TitleAuthorsApplication AreaTherapeutic AreaJournalYearLinkPageVolumeIssuePubMed IDAbstractImpact FactorSnippetDateDOI
A benzothiophene inhibitor of mitogen-activated protein kinase-activated protein kinase 2 inhibits tumor necrosis factor alpha production and has oral anti-inflammatory efficacy in acute and chronic models of inflammationMourey, RJ;Burnette, BL;Brustkern, SJ;Daniels, JS;Hirsch, JL;Hood, WF;Meyers, MJ;Mnich, SJ;Pierce, BS;Saabye, MJ;Schindler, JF;South, SA;Webb, EG;Zhang, J;Anderson, DR;Pharmacokinetics (PK)Inflammatory DiseaseJ. Pharmacol. Exp. Ther.2010http://jpet.aspetjournals.org/content/333/3/797.short797-807333320237073Activation of the p38 kinase pathway in immune cells leads to the transcriptional and translational regulation of proinflammatory cytokines. Mitogen-activated protein kinase-activated protein kinase 2 (MK2), a direct downstream substrate of p38 kinase, regulates lipopolysaccharide (LPS)-stimulated tumor necrosis factor alpha (TNFalpha) and interleukin-6 (IL-6) production through modulating the stability and translation of these mRNAs. Developing small-molecule inhibitors of MK2 may yield anti-inflammatory efficacy with a different safety profile relative to p38 kinase inhibitors. This article describes the pharmacologic properties of a benzothiophene MK2 inhibitor, PF-3644022 [(10R)-10-methyl-3-(6-methylpyridin-3-yl)-9,10,11,12-tetrahydro-8H-[1,4]diazepino[5',6':4,5]thieno[3,2-f]quinolin-8-one]. PF-3644022 is a potent freely reversible ATP-competitive compound that inhibits MK2 activity (K(i) = 3 nM) with good selectivity when profiled against 200 human kinases. In the human U937 monocytic cell line or peripheral blood mononuclear cells, PF-3644022 potently inhibits TNFalpha production with similar activity (IC(50) = 160 nM). PF-3644022 blocks TNFalpha and IL-6 production in LPS-stimulated human whole blood with IC(50) values of 1.6 and 10.3 microM, respectively. Inhibition of TNFalpha in U937 cells and blood correlates closely with inhibition of phospho-heat shock protein 27, a target biomarker of MK2 activity. PF-3644022 displays good pharmacokinetic parameters in rats and is orally efficacious in both the rat acute LPS-induced TNFalpha model and the chronic streptococcal cell wall-induced arthritis model. Dose-dependent inhibition of TNFalpha production in the acute model and inhibition of paw swelling in the chronic model is observed with ED(50) values of 6.9 and 20 mg/kg, respectively. PF-3644022 efficacy in the chronic inflammation model is strongly correlated with maintaining a C(min) higher than the EC(50) measured in the rat LPS-induced TNFalpha model.3.615Male Sprague-Dawley rats weighing 275 to 300 g were purchased from Charles River Laboratories Inc. (Wilmington, MA) and acclimated to their surroundings for approximately 1 week with food and water provided ad libitum. A minimum of 1 day before study, animals were anesthetized with isoflurane (to effect) and then implanted with Culex (BASi, West Lafayette, IN) vascular catheters in the carotid artery. Animals were acclimated in Culex cages overnight before dosing. Patency of the carotid artery catheter was maintained by using the “tend” function of Culex ABS. PF-3644022 was administered dissolved in 70% normal saline/20% polyethylene glycol-400/10% ethanol (intravenously) or suspended in 0.5% hydroxypropylmethylcellulose/0.1% Tween 80 in distilled water (by mouth). Blood collections were obtained from the carotid artery and performed by the Culex at 2 (intravenous only), 5, 15, 30 min and 1, 2, 4, 6, 82010-06-01 00:00:0010.1124/jpet.110.166173
A nonspecific phosphotyrosine phosphatase inhibitor, bis(maltolato)oxovanadium(IV), improves glucose tolerance and prevents diabetes in Zucker diabetic fatty ratsWinter, CL;Lange, JS;Davis, MG;Gerwe, GS;Downs, TR;Peters, KG;Kasibhatla, B;Glucose Tolerance TestDiabetesExp. Biol. Med. (Maywood)2005https://journals.sagepub.com/doi/abs/10.1177/153537020523000307207-216230315734724The molecular basis of insulin resistance, a major risk factor for development of Type II diabetes, involves defective insulin signaling. Insulin-mediated signal transduction is negatively regulated by the phosphotyrosine phosphatase, PTP1B, and numerous studies have demonstrated that organo-vanadium compounds, which are nonselective phosphotyrosine phosphatase inhibitors, have insulin-mimetic properties. However, whether or not vanadium compounds can prevent the transition from insulin resistance to overt diabetes is unknown. We compared the ability of bis(maltolato)oxovanadium(IV) (BMOV), an orally bioavailable organo-vanadium compound, and rosiglitazone maleate (RSG), a known insulin sensitizer, to prevent development of diabetes in Zucker diabetic fatty (ZDF) rats. Treatment began at 6 weeks of age when animals are insulin resistant and hyperinsulinemic, but not yet hyperglycemic, and ended at 12 weeks of age, which is 4 weeks after ZDF rats typically develop overt diabetes. BMOV-treated ZDF rats did not develop hyperglycemia, showed significant improvement in insulin sensitivity, and retained normal pancreatic islet morphology and endocrine cell distribution, similar to RSG-treated animals. BMOV and RSG treatment also prevented the hyper-phagia and polydipsia present in untreated ZDF rats; however, BMOV-treated ZDF rats gained much less weight than did RSG-treated animals. Circulating levels of adiponectin decreased in untreated ZDF rats compared to lean controls, but these levels remained normal in BMOV-treated ZDF rats. In contrast, in RSG-treated ZDF rats, plasma adiponectin levels were nearly 4-fold higher than in lean control rats, primarily as a result of a large increase in the amount of low-molecular weight forms of adiponectin in circulation. These data demonstrate that phosphatase inhibition offers a new approach to diabetes prevention, one that may have advantages over current approaches.At the end of the 6-week treatment period, a jugular vein catheter was surgically implanted, and the rats were allowed to recover for 24 hrs. Following a 16-hr fast, an oral glucose tolerance test was performed. Glucose was administered via oral gavage at a dose of 2 g/kg of body weight. Blood was sampled at 0, 30, 60,90, and 120 min using a Culex automated blood sampler from Bioanalytical Systems (West Lafayette, IN). The blOod was collected in sodium heparin tubes, centrifuged, and the plasma was analyzed for glucose levels using a Yellow Springs, Inc. (YSI) 2700 (Yellow Springs, OH) blOod glucose analyzer. The remaining plasma was frozen at -80°C for insulin analysis2005-03-01 00:00:0010.1177/153537020523000307
A novel autotaxin inhibitor reduces lysophosphatidic acid levels in plasma and the site of inflammationGierse, J;Thorarensen, A;Beltey, K;Bradshaw-Pierce, E;Cortes-Burgos, L;Hall, T;Johnston, A;Murphy, M;Nemirovskiy, O;Ogawa, S;Pegg, L;Pelc, M;Prinsen, M;Schnute, M;Wendling, J;Wene, S;Weinberg, R;Wittwer, A;Zweifel, B;Masferrer, J;PK/PDInflammatory DiseaseJ. Pharmacol. Exp. Ther.2010http://jpet.aspetjournals.org/content/334/1/310.short310-317334120392816Autotaxin is the enzyme responsible for the production of lysophosphatidic acid (LPA) from lysophosphatidyl choline (LPC), and it is up-regulated in many inflammatory conditions, including but not limited to cancer, arthritis, and multiple sclerosis. LPA signaling causes angiogenesis, mitosis, cell proliferation, and cytokine secretion. Inhibition of autotaxin may have anti-inflammatory properties in a variety of diseases; however, this hypothesis has not been tested pharmacologically because of the lack of potent inhibitors. Here, we report the development of a potent autotaxin inhibitor, PF-8380 [6-(3-(piperazin-1-yl)propanoyl)benzo[d]oxazol-2(3H)-one] with an IC(50) of 2.8 nM in isolated enzyme assay and 101 nM in human whole blood. PF-8380 has adequate oral bioavailability and exposures required for in vivo testing of autotaxin inhibition. Autotaxin's role in producing LPA in plasma and at the site of inflammation was tested in a rat air pouch model. The specific inhibitor PF-8380, dosed orally at 30 mg/kg, provided >95% reduction in both plasma and air pouch LPA within 3 h, indicating autotaxin is a major source of LPA during inflammation. At 30 mg/kg PF-8380 reduced inflammatory hyperalgesia with the same efficacy as 30 mg/kg naproxen. Inhibition of plasma autotaxin activity correlated with inhibition of autotaxin at the site of inflammation and in ex vivo whole blood. Furthermore, a close pharmacokinetic/pharmacodynamic relationship was observed, which suggests that LPA is rapidly formed and degraded in vivo. PF-8380 can serve as a tool compound for elucidating LPA's role in inflammation.3.615Male Lewis rats weighing 275 to 300 g were purchased from Charles River Laboratories, Inc. (Wilmington, MA) and acclimated to their surroundings for approximately 1 week with food and water provided ad libitum. A minimum of 1 day before study, animals were anesthetized with isoflurane (to effect) and implanted with Culex (BASi, West Lafayette, IN) vascular catheters in the carotid artery. Animals were acclimated in Culex cages overnight before dosing. Patency of the carotid artery catheter was maintained by using the “tend” function of the Culex automated blood sampler. Animals were dosed with PF-8380 at 1, 3, 10, 30, and 100 mg/kg by oral gavage after an overnight fast. Blood collections were obtained from the carotid artery and performed by the Culex automated blood sampler at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 h after administration. Blood was centrifuged, and plasma was collected for analysis of PF-8380 and LPA concentrations2010-07-01 00:00:0010.1124/jpet.110.165845
AMP-activated protein kinase signaling activation by resveratrol modulates amyloid-beta peptide metabolismVingtdeux, V;Giliberto, L;Zhao, H;Chandakkar, P;Wu, Q;Simon, JE;Janle, EM;Lobo, J;Ferruzzi, MG;Davies, P;Marambaud, P;PK/PDAlzheimersJ. Biol. Chem.2010http://www.jbc.org/content/285/12/9100.short9100-91132851220080969Alzheimer disease is an age-related neurodegenerative disorder characterized by amyloid-beta (Abeta) peptide deposition into cerebral amyloid plaques. The natural polyphenol resveratrol promotes anti-aging pathways via the activation of several metabolic sensors, including the AMP-activated protein kinase (AMPK). Resveratrol also lowers Abeta levels in cell lines; however, the underlying mechanism responsible for this effect is largely unknown. Moreover, the bioavailability of resveratrol in the brain remains uncertain. Here we show that AMPK signaling controls Abeta metabolism and mediates the anti-amyloidogenic effect of resveratrol in non-neuronal and neuronal cells, including in mouse primary neurons. Resveratrol increased cytosolic calcium levels and promoted AMPK activation by the calcium/calmodulin-dependent protein kinase kinase-beta. Direct pharmacological and genetic activation of AMPK lowered extracellular Abeta accumulation, whereas AMPK inhibition reduced the effect of resveratrol on Abeta levels. Furthermore, resveratrol inhibited the AMPK target mTOR (mammalian target of rapamycin) to trigger autophagy and lysosomal degradation of Abeta. Finally, orally administered resveratrol in mice was detected in the brain where it activated AMPK and reduced cerebral Abeta levels and deposition in the cortex. These data suggest that resveratrol and pharmacological activation of AMPK have therapeutic potential against Alzheimer disease.4.125Resveratrol Pharmacokinetics and Brain Accumulation in Rodents— All animal procedures were approved by the Purdue Animal Care and Use Committee. Sprague-Dawley rats were dosed orally by gavage with resveratrol in a dose escalation schedule and after an oral 400- mg/kg dose. Blood sampling was obtained in the CulexTM automated system (Bioanalytical Systems, West Lafayette, IN). Rats were then fasted for 8 h, dosed orally with 400 mg/kg resveratrol, and sacrificed 1 h postdose2010-03-19 00:00:00
An acute rat in vivo screening model to predict compounds that alter blood glucose and/or insulin regulationBrott, DA;Diamond, M;Campbell, P;Zuvich, A;Cheatham, L;Bentley, P;Gorko, MA;Fikes, J;Saye, J;Glucose Tolerance TestDiabetesJ Pharmacol Toxicol Methods2013https://www.sciencedirect.com/science/article/pii/S105687191300275X190-196682Drug-induced glucose dysregulation and insulin resistance have been associated with weight gain and potential induction and/or exacerbation of diabetes mellitus in the clinic suggesting they may be safety biomarkers when developing antipsychotics. Glucose and insulin have also been suggested as potential efficacy biomarkers for some oncology compounds. The objective of this study was to qualify a medium throughput rat _in vivo_ acute Intravenous Glucose Tolerance Test (IVGTT) for predicting compounds that will induce altered blood glucose and/or insulin levels.2.679Cannulas were externalized at the nape of the neck, and the animals allowed at least 4 days of recovery, prior to the IVGTT. The day prior to the experiment, rats were jacketed, tethered and housed individually in the Culex® (BASi, West Lafayette, IN) system ;and 2) clozapine increased the basal glucose level in the male mouse (Dwyer & compound risk for inducing clinical weight gain, 3) determine if different rat strains/substrains experiment, rats were jacketed, tethered and housed individually in the Culex® (BASi, West Lafayette 2013-07-05 00:00:0010.1016/j.vascn.2013.06.003
An automated blood sampler for simultaneous sampling of systemic blood and brain microdialysates for drug absorption, distribution, metabolism, and elimination studiesGunaratna, PC;Kissinger, PT;Kissinger, CB;Gitzen, JF;ADMEMethod DevelopmentJ Pharmacol Toxicol Methods2004https://www.sciencedirect.com/science/article/pii/S105687190300058357-6449114670694A major problem in preclinical drug development where blood sampling from small animals is a routine practice is the time and labor involved in the serial sampling of small blood volumes from small animals such as rats for the duration of pharmacokinetic/pharmacodynamic (PK/PD) studies. The traditional method of manually drawing blood from the animal requires the animal to be anesthetized or restrained with some device, both of which cause stress to the animal. An automated blood sampler (ABS) was developed to simultaneously collect blood and brain microdialysate samples at preprogrammed time points from awake and freely moving animals. The samples are delivered to fraction collectors and stored at 4 degrees C until use. The lost blood volume during collection is replaced with sterile saline to prevent fluid loss from the animal. In addition, the system is capable of collecting urine and feces for metabolism studies and monitoring the animal activity for behavioral studies. In the present study, blood samples were collected for 24 h after dosing rats orally with a 5 mg/kg dose of olanzapine (OLAN). Brain dialysates were collected for the same duration from a microdialysis probe implanted in the striatum. The pharmacokinetic parameters, obtained after an oral dose, are in good agreement with reported values in literature. The pharmacodynamic information obtained from brain dialysates data show that OLAN elevates the concentration of dopamine (DA) in the brain and remains in the brain even after it is cleared from the plasma. The ABS described here is a very useful tool in drug development to accelerate the pace of preclinical in vivo studies and to simultaneously provide pharmacodynamic and physiological information.2.679and a BAS Sample Sentinel autosampler with a 20-μl injection loop (Bioanalytical Systems, West Lafayette During the 24-h postsurgery recovery period in the Culex ABS, the catheter patency was temperature on a microbore ODS column (150×1.0 mm, 3 μm) (BAS Unijet) using 2004-12-13 00:00:0010.1016/S1056-8719(03)00058-3
An initial investigation into the effects of isolation and enrichment on the welfare of laboratory pigs housed in the PigTurn® system, assessed using tear staining, behaviour, physiology and haematologyDeBoer, S;Garner, J;McCain, R;Lay Jr, D;Eicher, S;Marchant-Forde, J;Stress HormonesMethod Developmentanim welf2015http://dx.doi.org/10.7120/09627286.24.1.01515-272411.5222-m catheter extension fixed to the automatic sampling system (Culex-L, BASi, West Lafayette 1 ml of heparinised (10 units ml-1) saline into the pig to maintain Automated blood collection was controlled by an automated sampling system (Culex, BioAnalytical Systems Inc, West2015-02-15 00:00:0010.7120/09627286.24.1.015
An integrative pharmacological approach to radio telemetry and blood sampling in pharmaceutical drug discovery and safety assessmentLitwin, DC;Lengel, DJ;Kamendi, HW;Bialecki, RA;Safety PharmacologyMethod DevelopmentBiomed Eng Online2011https://biomedical-engineering-online.biomedcentral.com/articles/10.1186/1475-925X-10-551021244682A successful integration of the automated blood sampling (ABS) and telemetry (ABST) system is described. The new ABST system facilitates concomitant collection of physiological variables with blood and urine samples for determination of drug concentrations and other biochemical measures in the same rat without handling artifact. Integration was achieved by designing a 13 inch circular receiving antenna that operates as a plug-in replacement for the existing pair of DSI's orthogonal antennas which is compatible with the rotating cage and open floor design of the BASi Culex® ABS system. The circular receiving antenna's electrical configuration consists of a pair of electrically orthogonal half-toroids that reinforce reception of a dipole transmitter operating within the coil's interior while reducing both external noise pickup and interference from other adjacent dipole transmitters. For validation, measured baclofen concentration (ABST vs. satellite (μM): 69.6 ± 23.8 vs. 76.6 ± 19.5, p = NS) and mean arterial pressure (ABST vs. traditional DSI telemetry (mm Hg): 150 ± 5 vs.147 ± 4, p = NS) variables were quantitatively and qualitatively similar between rats housed in the ABST system and traditional home cage approaches. The ABST system offers unique advantages over traditional between-group study paradigms that include improved data quality and significantly reduced animal use. The superior within-group model facilitates assessment of multiple physiological and biochemical responses to test compounds in the same animal. The ABST also provides opportunities to evaluate temporal relations between parameters and to investigate anomalous outlier events because drug concentrations, physiological and biochemical measures for each animal are available for comparisons.2.013Methods. Reasons for integration of the BASi Culex ® and DSI radio telemetry Paul, MN) and the Culex ® automated blood sampling (ABS) system (BAS Inc, West Lafayette, IN) The Culex ® system can accommodate awake and freely moving animals such as the rat2011-01-18 00:00:0010.1186/1475-925X-10-5
Application of automated serial blood sampling and dried blood spot technique with liquid chromatography-tandem mass spectrometry for pharmacokinetic studies in miceWong, P;Pham, R;Whitely, C;Soto, M;Salyers, K;James, C;Bruenner, BA;DBSMethod DevelopmentJ Pharm Biomed Anal2011https://www.sciencedirect.com/science/article/pii/S0731708511003426604-60856321784595The goal of this work was to obtain full pharmacokinetic profiles from individual mice with the use of an automated blood sampling system and dried blood spot (DBS) technique. AMG 517, a potent and selective vanilloid receptor (VR1) antagonist, was dosed to mice (n=3) intravenously and blood samples were collected using the automated blood sampling system with the "no blood waste" method. The collected blood samples were a mixture of 25 μL blood and 50 μL of heparinized saline solution. Two 15 μL aliquots were manually spotted onto a DBS card and dried at room temperature for at least 2h before being stored in zip bags with desiccant. The remaining samples (45 μL) were stored at -70°C until analysis. Both the DBS and the whole blood samples (diluted with saline (1:2, v/v)) were extracted and analyzed by liquid chromatography-tandem mass spectrometry. The overall extraction recovery of the analyte from the dried blood spots was determined to be about 90%. The pharmacokinetic parameters calculated using the whole blood or the DBS concentration data were comparable, and were obtained from only 3 mice, whereas conventional sampling and analysis would have required up to 27 mice to achieve the same result. The analyte was shown to be stable in the diluted whole blood (blood:saline 1:2) at room temperature for at least 4h and in the DBS for at least 34 days when stored at room temperature. These results indicated that the automated blood sampling system and DBS collection are promising techniques to obtain full pharmacokinetic profiles from individual mice and reduce the use of animals. Copyright © 2011. Published by Elsevier B.V.2.983A 30 μL aliquot of 100 μg/mL AMG 517 was spiked into 570 μL of mouse whole blood to obtain a 5000 ng/mL mixed with heparinized saline solution in a ratio of 1:2 (v/v) to mimic the collected blood samples from the Culex ® system (Bioanalytical Systems, West Lafayette ; A 30 μL aliquot of 100 μg/mL AMG 517 was spiked into 570 μL of mouse whole blood to obtain a 5000 ng/mL mixed with heparinized saline solution in a ratio of 1:2 (v/v) to mimic the collected blood samples from the Culex ® system (Bioanalytical Systems, West Lafayette ; A 30 μL aliquot of 100 μg/mL AMG 517 was spiked into 570 μL of mouse whole blood to obtain a 5000 ng/mL mixed with heparinized saline solution in a ratio of 1:2 (v/v) to mimic the collected blood samples from the Culex ® system (Bioanalytical Systems, West Lafayette ; Acknowledgements. The authors would like to thank Candace Rohde and Scott Peters from Bioanalytical Systems (West Lafayette, IN) for technical support of the Culex ® automated blood sampling system. Recommended articles Citing articles (0). References ; A 30 μL aliquot of 100 μg/mL AMG 517 was spiked into 570 μL of mouse whole blood to obtain a 5000 ng/mL mixed with heparinized saline solution in a ratio of 1:2 (v/v) to mimic the collected blood samples from the Culex ® system (Bioanalytical Systems, West Lafayette2011-11-01 00:00:0010.1016/j.jpba.2011.06.022
Assessment of a Maslinic Acid Derivative and its Metabolite in Rat Blood by Liquid Chromatography Coupled with Mass SpectrometryPavel, IZ;Dehelean, CA;Farczadi, L;ADMENutritional SupplementsThesis2017https://pdfs.semanticscholar.org/02c9/a2696a550f0c05fe9fefb96de168e0404fc6.pdfKeywords: LC/MS analysis, maslinic acid derivative, metabolite, rat blood The maslinic acid derivative (50 mg/kg of body weight) was given via the oral and intraperitoneal route [3, 7]. Blood was automatically collected using the Culex System [BASi Corporate, Indiana2017-01-01 00:00:00
Assessment of cisplatin-induced kidney injury using an integrated rodent platformChen, Y;Brott, D;Luo, W;Gangl, E;Kamendi, H;Barthlow, H;Lengel, D;Fikes, J;Kinter, L;Valentin, JP;Bialecki, R;Safety PharmacologyRenal InjuryToxicol. Appl. Pharmacol.2013https://www.sciencedirect.com/science/article/pii/S0041008X1300063X352-361268323415679Current diagnosis of drug-induced kidney injury (DIKI) primarily relies on detection of elevated plasma creatinine (Cr) or blood urea nitrogen (BUN) levels; however, both are indices of overall kidney function and changes are delayed with respect to onset of nephron injury. Our aim was to investigate whether early changes in new urinary DIKI biomarkers predict plasma Cr, BUN, renal hemodynamic and kidney morphological changes associated with kidney injury following a single dose of cisplatin (CDDP) using an integrated platform in rodent. Conscious surgically prepared male Han Wistar rats were given a single intraperitoneal dose of CDDP (15mg/kg). Glomerular filtration rate (GFR), effective renal plasma flow (ERPF), urinalysis, DIKI biomarkers, CDDP pharmacokinetics, blood pressures, heart rate, body temperature and electroencephalogram (EEG) were measured in the same vehicle- or CDDP-treated animals over 72h. Plasma chemistry (including Cr and BUN) and renal tissues were examined at study termination. Cisplatin caused progressive reductions of GFR, ERPF, heart rate and body temperature from day 1 (0-24h). DIKI biomarkers including alpha-glutathione S-transferase (α-GST) significantly increased as early as 6h post-dose, which preceded significant declines of GFR and ERPF (24h), increased plasma Cr and BUN (72h), and associated with renal acute tubular necrosis at 72h post-dose. The present study adds to the current understanding of CDDP action by demonstrating that early increases in urinary excretion of α-GST predict DIKI risk following acute exposure to CDDP in rats, before changes in traditional DIKI markers are evident. Copyright © 2013 Elsevier Inc. All rights reserved.3.585Animals were allowed free access to bottle water (sterilized tap water) and standard rat chow throughout the duration of the study. ABST system and data acquisition. The integrated pharmacology testing platform (Fig. 1) consisted of the BASi Culex® automated blood sampling2013-05-01 00:00:0010.1016/j.taap.2013.01.032
Automated method development of sample preparationSingleton, C;Li, M;Pharmacokinetics (PK)Data ReviewEliminating Bottlenecks for Efficient Bioanalysis: Practices and Applications in Drug Discovery and Development2014https://www.futuremedicine.com/doi/full/10.4155/fseb2013.13.396-17The automation of sample preparation has been underway for many years now, with reliable and robust assays being developed for most types of typical extraction procedures. On the horizon is the automation of method development for sample preparation, which has great potential to streamline workflows and routine bench work commonly encountered in a bioanalytical laboratory. This approach is still in its infancy, and dedicated development and refinement staff are recommended for successful application. Herein, we present recent work in the automation of the method development process as well as some guidelines for implementing this approach in the laboratory.One can envision a scenario where an automated sample collection system such as a Culex® (BASi, IN, USA) draws sample at set intervals and dispenses them [17]. Then a user receives the samples and generates an Page 102014-10-01 00:00:0010.4155/fseb2013.13.39
Behavioral control over shock blocks behavioral and neurochemical effects of later social defeatAmat, J;Aleksejev, RM;Paul, E;Watkins, LR;Maier, SF;MicrodialysisCNS StressNeuroscience2010https://www.sciencedirect.com/science/article/pii/S03064522090182231031-1038165419909791Experience with behavioral control over tailshock (escapable shock, ES) has been shown to block the behavioral and neurochemical changes produced by later uncontrollable tail shock (inescapable shock, IS). The present experiments tested, in rats, whether the protective effect of control over tailshock extends beyond reducing the behavioral and neurochemical impact of a subsequent tailshock experience to stressors that are quite different. Social defeat (SD) was chosen as the second stress experience because it has few if any cues in common with tailshock. SD produced shuttlebox escape learning deficits ("learned helplessness") and reduced juvenile social investigation 24 h later, as does IS. IS is notable for inducing a large increase in dorsal raphe nucleus (DRN) serotonergic (5-HT) activity as measured by extracellular levels of 5-HT within the DRN, and SD did so as well. ES occurring 7 days before SD blocked this SD-induced DRN activation, as well as the SD-induced interference with shuttlebox escape and reduction in social investigation. Prior exposure to yoked IS did not reduce the DRN 5-HT activation or later behavioral effects produced by SD, and thus the proactive stress-blunting effects of ES can be attributed to it's controllability. Thus, ES confers a very general protection to the impact of a subsequent stress experience.3.244Each animal was placed individually in a Plexiglas bowl (Bioanalytical Systems, West Lafayette, IN, USA) and increases extracellular levels of 5-HT in the dorsal raphe nucleus of the rat of repellent action of neem oil against the filarial vector, Culex quinquefasciatus (Diptera2010-02-17 00:00:0010.1016/j.neuroscience.2009.11.005
Bioanalytical method development and validation of corynantheidine, a kratom alkaloid, using UPLC-MS/MS, and its application to preclinical pharmacokinetic studiesKing, TI;Sharma, A;Kamble, SH;León, F;Berthold, EC;Popa, R;Cerlati, O;Prentice, BM;McMahon, LR;McCurdy, CR;Avery, BA;PK/PDAddictionJ Pharm Biomed Anal2020https://www.sciencedirect.com/science/article/pii/S073170851932291511301918031838282Corynantheidine, a minor alkaloid found in Mitragyna speciosa (Korth.) Havil, has been shown to bind to opioid receptors and act as a functional opioid antagonist, but its unique contribution to the overall properties of kratom remains relatively unexplored. The first validated bioanalytical method for the quantification of corynantheidine in rat plasma is described. The method was linear in the dynamic range from 1-500 ng/mL, requires a small plasma sample volume (25 μL), and a simple protein precipitation method for extraction of the analyte. The separation was achieved with Waters BEH C18 2.1 × 50 mm column and the 3-minute gradient of 10 mM ammonium acetate buffer (pH = 3.5) and acetonitrile as mobile phase. The method was validated in terms of accuracy, precision, selectivity, sensitivity, recovery, stability, and dilution integrity. It was applied to the analysis of the male Sprague Dawley rat plasma samples obtained during pharmacokinetic studies of corynantheidine administered both intravenously (I.V.) and orally (P.O.) (2.5 mg/kg and 20 mg/kg, respectively). The non-compartmental analysis performed in Certara Phoenix® yielded the following parameters: clearance 884.1 ± 32.3 mL/h, apparent volume of distribution 8.0 ± 1.2 L, exposure up to the last measured time point 640.3 ± 24.0 h*ng/mL, and a mean residence time of 3.0 ± 0.2 h with I.V. dose. The maximum observed concentration after a P.O. dose of 213.4 ± 40.4 ng/mL was detected at 4.1 ± 1.3 h with a mean residence time of 8.8 ± 1.8 h. Absolute oral bioavailability was 49.9 ± 16.4 %. Corynantheidine demonstrated adequate oral bioavailability, prolonged absorption and exposure, and an extensive extravascular distribution. In addition, imaging mass spectrometry analysis of the brain tissue was performed to evaluate the distribution of the compound in the brain. Corynantheidine was detected in the corpus callosum and some regions of the hippocampus. Copyright © 2019 Elsevier B.V. All rights reserved.2.983blood collection vials and sterile blood collection tubing were obtained from BASi (West Lafayette The high concentration plasma (HCP) was then diluted with blank rat plasma (BRP) at 5 the duration of the PK studies, animals were contained inside the Culex® automated blood2020-02-20 00:00:0010.1016/j.jpba.2019.113019
Bioanalytical method development and validation of MES207, a neuropeptide FF receptor antagonist, and its application in preclinical pharmacokineticsKing, TI;Roewekamp, AC;Sharma, A;Harrison, S;Mesangeau, C;Mottinelli, M;Kamble, SH;McCurdy, CR;Avery, BA;Pharmacokinetics (PK)Method DevelopmentJ. Chromatogr. B Analyt. Technol. Biomed. Life Sci.2019https://www.sciencedirect.com/science/article/pii/S15700232193122921218751134-113531790916The nonpeptide small molecule, MES207, exhibits 17-fold preferential binding to the neuropeptide FF receptor 1 (NPFFR1) over NPFFR2 and shows antagonist functionality at NPFF receptors. In order to further the development of MES207 as a NPFFR1 probe, an UPLC-MS/MS bioanalytical method was developed and validated to quantify MES207 in rat plasma for a linearity range of 3-200 ng/mL. The method was applied in the analysis of the plasma, brain, and urine samples collected during pharmacokinetic studies in healthy male and female Sprague Dawley rats. The animals were dosed through oral gavage (50 mg/kg) and intravenously (2.5 mg/kg). Test samples were analyzed using the validated bioanalytical method to generate plasma concentration-time profiles. The results were further subjected to non-compartmental analysis using Phoenix 6.4®. MES207 exhibits a large volume of distribution (1.2 ± 0.6 L), high clearance (0.8 ± 0.1 L/h), and a poor oral bioavailability (1.7 ± 0.4%). The compound also showed a multiple peak phenomenon with a very short absorption phase. It appears that gender does not significantly influence the differences in pharmacokinetic parameters of this NPFF probe. Copyright © 2019 Elsevier B.V. All rights reserved.2.813the duration of the experiment, animals were housed in individual single occupancy BASi Culex® (West Lafayette All metabolic cages (Nalgene and Culex) consisted of mesh floor, collecting funnel for urine, and a single oral dose of 50 mg/kg was given to a conscious rat by oral2019-12-15 00:00:0010.1016/j.jchromb.2019.121875
Biochemical, cellular, and anti-inflammatory properties of a potent, selective, orally bioavailable benzamide inhibitor of Rho kinase activityRajagopalan, LE;Davies, MS;Kahn, LE;Kornmeier, CM;Shimada, H;Steiner, TA;Zweifel, BS;Wendling, JM;Payne, MA;Loeffler, RF;Case, BL;Norton, MB;Parikh, MD;Nemirovskiy, OV;Mourey, RJ;Masferrer, JL;Misko, TP;Kolodziej, SA;Pharmacokinetics (PK)Inflammatory DiseaseJ. Pharmacol. Exp. Ther.2010http://jpet.aspetjournals.org/content/333/3/707.short707-716333320228155Rho kinase, is the most widely studied downstream effector of the small Rho GTPase RhoA. Two Rho kinase isoforms have been described and are frequently referred to in the literature as ROCK1 and ROCK2. The RhoA-Rho kinase pathway has been implicated in the recruitment of cellular infiltrates to disease loci in a number of preclinical animal models of inflammatory disease. In this study, we used biochemical enzyme assays and a cellular target biomarker assay to define PF-4950834 [N-methyl-3-{[(4-pyridin-4-ylbenzoyl)amino]methyl}benzamide] as an ATP-competitive, selective Rho kinase inhibitor. We further used PF-4950834 to study the role of Rho kinase activation in lymphocyte and neutrophil migration in addition to the endothelial cell-mediated expression of adhesion molecules and chemokines, which are essential for leukocyte recruitment. The inhibitor blocked stromal cell-derived factor-1alpha-mediated chemotaxis of T lymphocytes in vitro and the synthesis of vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 in activated human endothelial cells in vitro. The secretion of chemokines interleukin-8 and monocyte chemoattractant protein-1 was also inhibited in activated endothelial cells. In addition, when dosed orally, the compound potently inhibited neutrophil migration in a carrageenan-induced acute inflammation model. In summary, we have used a pharmacologic approach to link Rho kinase activation to multiple phenotypes that can contribute to leukocyte infiltration. Inhibition of this pathway therefore could be strongly anti-inflammatory and provide therapeutic benefit in chronic inflammatory diseases.3.615Use of animals in these studies was reviewed and approved by the Pfizer Institutional Animal Care and Use Committee. The animal care and use program is fully accredited by the Association for Assessment and Accreditation of Laboratory Animal Care, International. Male Sprague-Dawley rats weighing 275 to 300 g were purchased from Charles River Laboratories, Inc. (Wilmington, MA). Before study, animals were anesthetized with Isoflurane (to effect) and then implanted with Culex (BASi, West Lafayette, IN) vascular catheters in the carotid artery. Animals were acclimated in Culex cages overnight before dosing. Patency of the carotid artery catheter was maintained by using the “tend” function of Culex ABS. PF-4950834 was administered dissolved in 70% normal saline/20% polyethylene glycol-400/105 ethanol (intravenously) or suspended in 0.5% hydroxypropylmethylcellulose/0.1% Tween 80 in distilled water (orally). Blood collections were obtained from the carotid artery and performed by the Culex at 2 (intravenously only), 5, 15, and 30 min and 1, 2, 4, 6, 8, 12, 18, and 24 h. Plasma was separated and frozen for analysis2010-06-01 00:00:0010.1124/jpet.110.166033
Bioequivalence of Two Intravenous Artesunate Products with Its Active Metabolite Following Single and Multiple InjectionsLi, Q;Xie, L;Melendez, V;Weina, P;Pharmacokinetics (PK)AntimalarialPharmaceuticals2011https://www.mdpi.com/1424-8247/4/1/138138-15341In animal species and humans, artesunate (AS) undergoes extensive and complex biotransformation to an active metabolite, dihydroartemisinin (DHA). The bioequivalence of two intravenous AS pharmaceutical products with 5% NaHCO3 (China Formulation) or 0.3 M PBS (WRAIR Formulation) was determined in rats in a two-formulation, two-period, and two-sequence crossover experimental design. Following single and multiple intravenous administrations, a series of blood samples was collected by using an automated blood sampler and drug concentrations were analyzed by LC-MS/MS. The 90% CI of the difference between the two intravenous formulations was contained within 80–125% of the geometric mean of pharmacokinetic parameters for AS and DHA in all animals dosed. Hematological effects were studied on days 1 and 3 after the final dosing, and a rapidly reversible hematological toxicity (significant reductions in reticulocyte levels) was seen in the peripheral blood of the rats treated with each formulation. The results showed that bioequivalence with the parent compound and active metabolite was fulfilled in the 82.3–117.7% ranges of all parameters (AUC0–t, Cmax, concentration average and degree of fluctuation) in the two-period and two-sequence crossover studies following single and repeated intravenous injections. For the metabolite, the equivalence was satisfied in most pharmacokinetic parameters tested due to the variability in the hydrolysis rate of AS to DHA. The WRAIR formulation of AS was considered to be bioequivalent to the Chinese formulation at steady-state according to the total drug exposure, in terms of both parent drug and active metabolite, rapidly reversal in reticulocyte decline, and extension of single and multiple administrations. Therefore, the parent drug and active metabolites should play similar important roles in the determination of efficacy and safety of the drug. View Full-Text [/1424-8247/4/1/138/htm] _Keywords: _ intravenous artesunate; dihydroartemisinin; bioequivalence; bioavailability; rats intravenous artesunate [/search?q=intravenous%20artesunate]; dihydroartemisinin [/search?q=dihydroartemisinin]; bioequivalence [/search?q=bioequivalence]; bioavailability [/search?q=bioavailability]; rats [/search?q=rats] This is an open access article distributed under the Creative Commons Attribution License [https://creativecommons.org/licenses/by-nc-sa/3.0/] [SciFeed] [/1424-8247/4/1/138/scifeed_display]As reported in other literatures and research, artesunate seemed to be a prodrug of DHA in our studies. The plasma concentrations of AS and its active metabolite, DHA, were simultaneously measured by using LC-MS/MS. An automated blood sampler (Culex ABS, Bioanalytical Systems, Inc. West Lafayette, IN) was used for rat blood sample collection. Twenty-four rats were randomized to daily intravenous injection of AS for three days in a two-formulation, two-sequence, and two-period study. The formulations were considered bioequivalent if the 90% confidence interval (CI) of the mean difference for each variable between formulations and periods were within 80% and 125% by calculated with WinNonlin software (WinNonlin 5.2, Pharsight Co. Mountain View, CA. USA)2011-01-07 00:00:0010.3390/ph4010138
Blood microsampling using capillaries for drug-exposure determination in early preclinical studies: a beneficial strategy to reduce blood sample volumesDillen, L;Loomans, T;Van de Perre, G;Versweyveld, D;Wuyts, K;de Zwart, L;Pharmacokinetics (PK)MicrosamplingBioanalysis2014https://www.future-science.com/doi/abs/10.4155/bio.13.286293-3066324471951Capillary microsampling (CMS) of blood with subsequent blood analysis offers a potential strategy to deal with increased demand to reduce blood sample volumes in animal discovery and preclinical studies. A generic approach is presented allowing PK analysis in 15 µl blood samples. CMS blood exposure data were compared with the traditional plasma exposure results in rats and dogs. Blood PK profiles obtained for two different compounds were in agreement with profiles obtained in plasma. From these studies ex vivo blood to plasma ratios were also obtained. In a mouse study, blood PK profiles that were obtained following automatic sampling overlay with the blood PK profiles obtained with CMS. CMS in 15 µl glass capillaries allows collection and handling of small and exact volumes of blood. Although CMS can also be applied for plasma collection, the full benefit is only achieved with blood collection and analysis.2.321The Culex ® ABC (automated blood collection) was from BASi (IN, USA Three animals were dosed and blood was collected with an automatic sampling device (Culex) via the jugular vein as an acceptable compromise between the 25 µl that has been used for rat studies versus2014-02-01 00:00:0010.4155/bio.13.286
Cathepsin K: a cysteine protease with unique kinin-degrading propertiesGodat, E;Lecaille, F;Desmazes, C;Duchêne, S;Weidauer, E;Saftig, P;Brömme, D;Vandier, C;Lalmanach, G;PK and MicrodialysisBlood Brain Barrier (BBB)Biochem. J.2004http://www.biotechnocentre.fr/files/04-actes_colloque.pdf501-506383Pt. 315265002Taking into account a previous report of an unidentified enzyme from macrophages acting as a kininase, the ability of cysteine proteases to degrade kinins has been investigated. Wild-type fibroblast lysates from mice, by contrast with cathepsin K-deficient lysates, hydrolysed BK (bradykinin), and released two metabolites, BK-(1-4) and BK-(5-9). Cathepsin K, but not cathepsins B, H, L and S, cleaved kinins at the Gly4-Phe5 bond and the bradykinin-mimicking substrate Abz (o-aminobenzoic acid)-RPPGFSPFR-3-NO2-Tyr (3-nitrotyrosine) more efficiently (pH 6.0: kcat/K(m)=12500 mM(-1) x s(-1); pH 7.4: kcat/K(m)=6930 mM(-1) x s(-1)) than angiotensin-converting enzyme hydrolysed BK. Conversely Abz-RPPGFSPFR-3-NO2-Tyr was not cleaved by the Y67L (Tyr67-->Leu)/L205A (Leu205-->Ala) cathepsin K mutant, indicating that kinin degradation mostly depends on the S2 substrate specificity. Kininase activity was further evaluated on bronchial smooth muscles. BK, but not its metabolites BK(1-4) and BK(5-9), induced a dose-dependent contraction, which was abolished by Hoe140, a B2-type receptor antagonist. Cathepsin K impaired BK-dependent contraction of normal and chronic hypoxic rats, whereas cathepsins B and L did not. Taking together vasoactive properties of kinins and the potency of cathepsin K to modulate BK-dependent contraction of smooth muscles, the present data support the notion that cathepsin K may act as a kininase, a unique property among mammalian cysteine proteases.4.331Based on results already published on Blood Brain Barrier passage about morphine and morphine 6-β-D-glucuronide in rat. This experiment was carried out using Culex® and Raturn® to see whether an automatic sampling on awake, non stressed, freely moving animals was easy to set up. Animals were first submitted to vascular surgery in a femoral vein and to a probe implantation in the Striatum (AP: 1.20; L: 2; P:4). After one day to recover, 10 mg/kg morphine was administered via subcutaneous route. Infusion of Ringer like solution was carried out via syringe pump at a flow rate of 3 µL/min. 15 samples of 30 µL were taken over the 2.5 hour following the administration. Concomitantly, automatic blood sampling was carried out, and 11 samples of 50 µL of blood were taken. Blood was automatically replaced by the same volume of saline, while the total volume of blood collected did not exceeded 1.5 mL (less than the 10 % of total volume). An LC/MS-MS previously published was applied on these small volumes collected and the limit of sensitivity was highly improved using TSQ Quantum equipment2004-11-01 00:00:0010.1042/BJ20040864
Cebranopadol: a novel potent analgesic nociceptin/orphanin FQ peptide and opioid receptor agonistLinz, K;Christoph, T;Tzschentke, TM;Koch, T;Schiene, K;Gautrois, M;Schröder, W;Kögel, BY;Beier, H;Englberger, W;Schunk, S;De Vry, J;Jahnel, U;Frosch, S;Pharmacokinetics (PK)PainJ. Pharmacol. Exp. Ther.2014http://jpet.aspetjournals.org/content/349/3/535.short535-548349324713140Cebranopadol (trans-6'-fluoro-4',9'-dihydro-N,N-dimethyl-4-phenyl-spiro[cyclohexane-1,1'(3'H)-pyrano[3,4-b]indol]-4-amine) is a novel analgesic nociceptin/orphanin FQ peptide (NOP) and opioid receptor agonist [Ki (nM)/EC50 (nM)/relative efficacy (%): human NOP receptor 0.9/13.0/89; human mu-opioid peptide (MOP) receptor 0.7/1.2/104; human kappa-opioid peptide receptor 2.6/17/67; human delta-opioid peptide receptor 18/110/105]. Cebranopadol exhibits highly potent and efficacious antinociceptive and antihypersensitive effects in several rat models of acute and chronic pain (tail-flick, rheumatoid arthritis, bone cancer, spinal nerve ligation, diabetic neuropathy) with ED50 values of 0.5-5.6 µg/kg after intravenous and 25.1 µg/kg after oral administration. In comparison with selective MOP receptor agonists, cebranopadol was more potent in models of chronic neuropathic than acute nociceptive pain. Cebranopadol's duration of action is long (up to 7 hours after intravenous 12 µg/kg; >9 hours after oral 55 µg/kg in the rat tail-flick test). The antihypersensitive activity of cebranopadol in the spinal nerve ligation model was partially reversed by pretreatment with the selective NOP receptor antagonist J-113397[1-[(3R,4R)-1-cyclooctylmethyl-3-hydroxymethyl-4-piperidyl]-3-ethyl-1,3-dihydro-2H-benzimidazol-2-one] or the opioid receptor antagonist naloxone, indicating that both NOP and opioid receptor agonism are involved in this activity. Development of analgesic tolerance in the chronic constriction injury model was clearly delayed compared with that from an equianalgesic dose of morphine (complete tolerance on day 26 versus day 11, respectively). Unlike morphine, cebranopadol did not disrupt motor coordination and respiration at doses within and exceeding the analgesic dose range. Cebranopadol, by its combination of agonism at NOP and opioid receptors, affords highly potent and efficacious analgesia in various pain models with a favorable side effect profile.3.615The pharmacokinetic properties of cebranopadol in rats were investigated after a single intravenous dose of 160 µg/kg cebranopadol. The intravenous dose was administered as a bolus in a volume of 2 ml/kg with a catheter in the vena femoralis. Blood samples (200 µl/sample) were withdrawn via an implanted arterial catheter (arteria carotis) by an automated blood sampling system (Culex; Bioanalytical Systems Inc., West Lafayette, IN) at the following sampling times: 0 (predose), 5, 15, 30, 60, 180, 360, 720, and 1440 minutes after administration. Blood samples were centrifuged, and plasma was separated. Plasma concentrations of cebranopadol were determined using a validated liquid chromatography-tandem mass spectrometry method. The lower limit of quantification for cebranopadol in this method was 0.05 ng/ml using a sample volume of 50 µl of plasma2014-06-01 00:00:0010.1124/jpet.114.213694
Cellular impedance assays for predictive preclinical drug screening of kinase inhibitor cardiovascular toxicityLamore, SD;Kamendi, HW;Scott, CW;Dragan, YP;Peters, MF;Safety PharmacologyMethod DevelopmentToxicol. Sci.2013https://academic.oup.com/toxsci/article-abstract/135/2/402/1658334402-413135223897988Cardiovascular (CV) toxicity is a leading contributor to drug attrition. Implementing earlier testing has successfully reduced human Ether-à-go-go-Related Gene-related arrhythmias. How- ever, analogous assays targeting functional CV effects remain elusive. Demand to address this gap is particularly acute for kinase inhibitors (KIs) that suffer frequent CV toxicity. The drug class also presents some particularly challenging requirements for assessing functional CV toxicity. Specifically, an assay must sense a downstream response that integrates diverse kinase signaling pathways. In addition, sufficient throughput is essential for handling inherent KI nonselectivity. A new opportunity has emerged with cellular impedance technology, which detects spontaneous beating cardiomyocytes. Impedance assays sense morphology changes downstream of cardiomyocyte contraction. To evaluate cardiomyocyte impedance assays for KI screening, we investigated two distinct KI classes where CV toxicity was discovered late and target risks remain unresolved. Microtubule-associated protein/microtubule affinity regulating kinase (MARK) inhibitors decrease blood pressure in dogs, whereas checkpoint kinase (Chk) inhibitors (AZD7762, SCH900776) exhibit dose-limiting CV toxicities in clinical trials. These in vivo effects manifested in vitro as cardiomyocyte beat cessation. MARK effects were deemed mechanism associated because beat inhibition potencies correlated with kinase inhibition, and gene knockdown and microtubule-targeting agents suppressed beating. MARK inhibitor impedance and kinase potencies aligned with rat blood pressure effects. Chk inhibitor effects were judged off-target because Chk and beat inhibition potencies did not correlate and knockdowns did not alter beating. Taken together, the data demonstrate that cardiomyocyte impedance assays can address three unmet needs-detecting KI functional cardiotoxicity in vitro, determining mechanism of action, and supporting safety structure-activity relationships.Radio telemetry data acquisition. Two days before study, the rats were moved to the automated blood sampling and telemetry system (ABST), an integrated pharmacology testing platform consisting of the BASi Culex automated blood sampling apparatus (BASi, West Lafayette, IN) and DSI radio telemetry system (DSI, St Paul, MN) (Kamendi et al., 2010) to allow for simultaneous evaluation of functional radiotelemetry signals detected with the modified PhysioTel receiver adapted for the ABST (Litwin et al., 2011) and drug pharmacokinetic parameters. Sampling frequency for BP and temperature were set at 500 an2013-10-01 00:00:0010.1093/toxsci/kft167
Characterization of plasma cytokine response to intraperitoneally administered LPS & subdiaphragmatic branch vagus nerve stimulation in rat modelSomann, JP;Wasilczuk, KM;Neihouser, KV;Sturgis, J;Albors, GO;Robinson, JP;Powley, TL;Irazoqui, PP;Immune ResponseInflammatory DiseasePLoS ONE2019https://journals.plos.org/plosone/article/file?type=printable&id=10.1371/journal.pone.0214317e021431714330921373Vagus nerve stimulation (VNS) has been on the forefront of inflammatory disorder research and has yielded many promising results. Questions remain, however, about the biological mechanisms of such treatments and the inconsistencies in the methods used in research efforts. Here, we aimed to clarify the inflammatory response to intraperitoneal (IP) injections of lipopolysaccharide (LPS) in rats, while analyzing corresponding effects of electrical stimulation to subdiaphragmatic branches (anterior gastric, accessory celiac, and hepatic) of the left vagus nerve. We accomplished an in-depth characterization of the time-varying cytokine cascade response in the serum of 58 rats to an acute IP LPS challenge over a 330-minute period by utilizing curve-fitting and starting point-alignment methods. We then explored the post-LPS neuromodulation effects of electrically stimulating individually cuffed subdiaphragmatic branches. Through our analysis, we found there to be a consistent order of IP LPS cytokine response (IL-10, TNF-α, GM-CSF, IL-17F, IL-6, IL-22, INF-γ). Apart from IL-10, the IP cytokine cascade was more variable in starting time and occurred later than in previously recorded intravenous (IV) challenges. We also found distinct regulatory effects on multiple cytokine levels by each of the three subdiaphragmatic stimulation subsets. While the time-variability of IP LPS use in rats complicates its utility, we have shown it to be a practical, arguably more physiologically relevant method than IV in rats when our methods are used. More importantly, we have shown that selective subdiaphragmatic neurostimulation can be utilized to selectively induce specific effects on inflammation in the body.2.776Catheters for blood collection were placed before cuffing the vagus nerve branch. We followed the general procedures outline in the Culex surgical manual (Bioanalytical Systems, Inc.; West Lafayette, IN) for femoral cannulation in a rat [32]. Catheters in our experiment, however, were inserted into the femoral artery instead of the vein and remained exposed on the benchtop for connection to the Culex system versus being subcutaneously routed through the animal2019-03-28 00:00:0010.1371/journal.pone.0214317
Column liquid chromatography: equipment and instrumentationLaCourse, WR;Pharmacokinetics (PK)MicrosamplingAnal. Chem.2002https://pubs.acs.org/doi/abs/10.1021/ac020220q2813-28317412120906676.35Automation. He et al. (A54) investigated the feasibility of a Culex automated blood-sampling system in conjunction with LC/MS/MS analysis for the quantification of ketoconazole in rat, after an oral dose of 10 mg/kg of the test article. Powell and Tempst (A55) described a microflow-based instrument, consisting of multiple rotary valves, capillary tubing2002-06-15 00:00:0010.1021/ac020220q
Disposition and drug-drug interaction potential of veliparib (ABT-888), a novel and potent inhibitor of poly(ADP-ribose) polymeraseLi, X;Delzer, J;Voorman, R;de Morais, SM;Lao, Y;Pharmacokinetics (PK)OncologyDrug Metab. Dispos.2011http://dmd.aspetjournals.org/content/39/7/1161.short1161-116939721436403The disposition of veliparib [(R)-2-(2-methylpyrrolidin-2-yl)-1H-benzo[d]imidazole-4-carboxamide, ABT-888], a novel and potent inhibitor of poly(ADP-ribose) polymerase for the treatment of cancers, was investigated in rats and dogs after intravenous and oral administration of [(3)H]veliparib and compared with that of humans. Veliparib absorption was high. Dosed radioactivity was widely distributed in rat tissues. The majority of drug-related material was excreted in urine as unchanged drug (approximately 54, 41, and 70% of the dose in rats, dogs, and humans, respectively). A lactam M8 and an amino acid M3 were two major excretory metabolites in animals. In the circulation of animals and humans, veliparib was the major drug-related component, and M8 was one of the major metabolites. Monooxygenated metabolite M2 was significant in the rat and dog, and M3 was also significant in the dog. Veliparib biotransformation occurred on the pyrrolidine moiety via formation of a lactam, an amino acid, and an N-carbamoyl glucuronide, in addition to oxidation on benzoimidazole carboxamide and sequential glucuronidation. In vitro experiments using recombinant human cytochrome P450 (P450) enzymes identified CYP2D6 as the major enzyme metabolizing veliparib with minor contributions from CYP1A2, 2C19, and 3A4. Veliparib did not inhibit or induce the activities of major human P450s. Veliparib was a weak P-glycoprotein (P-gp) substrate, showing no P-gp inhibition. Taken together, these studies indicate a low potential for veliparib to cause clinically significant P-gp or P450-mediated drug-drug interactions (DDIs). Overall, the favorable dispositional and DDI profiles of veliparib should be beneficial to its safety and efficacy.3.354The cages were washed at the end of the study with a small amount of 70% ethanol in water, and the cage wash was collected for determination of total radioactivity. To obtain plasma samples for metabolite identification and profiling, a group of three surgically modified rats with femoral artery cannulas from Hilltop Laboratory Animals, Inc. was given a single 5 mg/kg intravenous dose of [3 H]veliparib via the superficial penile vein. Serial blood samples (0.5 ml at each time point) were taken via a Culex automated blood sampling system (BASi, West Lafayette, IN) from individual rats at 0.5, 1, 2, 4, 6, and 24 h postdose. Blood samples were centrifuged at 3000 rpm and 4°C to separate plasma (Jouan CR3 centrifuge, rotor model T40; Thermo Fisher Scientific, Waltham, MA), which were stored at 20°C until analysis2011-07-01 00:00:0010.1124/dmd.110.037820
Dose-Independent ADME Properties and Tentative Identification of Metabolites of α-Mangostin from Garcinia mangostana in Mice by Automated Microsampling and UPLC-MS/MS MethodsHan, SY;You, BH;Kim, YC;Chin, YW;Choi, YH;Pharmacokinetics (PK)Nutritional SupplementsPLoS ONE2015https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0131587e013158710726176540The information about a marker compound's pharmacokinetics in herbal products including the characteristics of absorption, distribution, metabolism, excretion (ADME) is closely related to the efficacy/toxicity. Also dose range and administration route are critical factors to determine the ADME profiles. Since the supply of a sufficient amount of a marker compound in in vivo study is still difficult, pharmacokinetic investigations which overcome the limit of blood collection in mice are desirable. Thus, we have attempted to investigate concurrently the ADME and proposed metabolite identification of α-mangostin, a major constituent of mangosteen, Garcinia mangostana L, in mice with a wide dose range using an in vitro as well as in vivo automated micro-sampling system together. α-mangostin showed dose-proportional pharmacokinetics at intravenous doses of 5-20 mg/kg and oral doses of 10-100 mg/kg. The gastrointestinal absorption of α-mangostin was poor and the distribution of α-mangostin was relatively high in the liver, intestine, kidney, fat, and lung. α-mangostin was extensively metabolized in the liver and intestine. With regards to the formation of metabolites, the glucuronidated, bis-glucuronidated, dehydrogenated, hydrogenated, oxidized, and methylated α-mangostins were tentatively identified. We suggest that these dose-independent pharmacokinetic characteristics of α-mangostin in mice provide an important basis for preclinical applications of α-mangostin as well as mangosteen. In addition, these experimental methods can be applied to evaluate the pharmacokinetics of natural products in mice.2.776The micro-sampling system (Culex BASi, West Lafayette, IN) was programmed to collect a 10 μL sample of blood into a micro-vial containing 50 μL of 12.5 units/mL heparinized saline. Blood loss due to blood sampling was replaced with equal volumes of heparinized saline ; Although the pharmacokinetic evaluation of α-MG was conducted using the plasma of rat [23], [24 The micro-sampling system (Culex BASi, West Lafayette, IN) was programmed to collect a 10 μL the urine collected over the previous 24 h. At this time, each mouse was sacrificed2015-07-15 00:00:0010.1371/journal.pone.0131587
Dried blood spots: concepts, present status, and future perspectives in bioanalysisSharma, A;Jaiswal, S;Shukla, M;Lal, J;DBSData ReviewDrug Test Anal2014https://onlinelibrary.wiley.com/doi/abs/10.1002/dta.1646399-4146524692095Over the past several years, dried blood spot (DBS) sampling technique has emerged as a pertinent method in both qualitative and quantitative bioanalysis context. In the DBS method, the blood sample is directly soaked on to a paper (with or without treatment). After drying it can be analyzed by modern analytical, immunological, or genomic detection systems. Several advantages of the DBS technique such as low blood volume requirement, transportation and storage without special treatment, better analytes stability, enhanced clinical cooperation in clinical trials, and reduced unforeseeable exposure of analysts to biohazards, make it the most appropriate blood sampling technique. This review illustrates the information available on the DBS method which may serve as a single window for investigators in the field of bioanalysis. Also, it explores the proficiency and appliance of the DBS method in pharmacokinetic (PK), therapeutic drug monitoring (TDM), toxicokinetic (TK), metabolomic, and disease diagnosis. Copyright © 2014 John Wiley & Sons, Ltd.2.799Online automated tools (ABS2; Instech Solomon, Plymouth Meeting, PA, USA and Culex; BASi, West Lafayette, IN, USA) are capable of collecting blood from freely moving laboratory animals and can be coupled for serial sampling (in microlitre of blood volume) on DBS cards ; For PK and TK studies in rat and mouse, blood can Online automated tools (ABS2; Instech Solomon, Plymouth Meeting, PA, USA and Culex; BASi, West Lafayette, IN, USA) are capable of collecting blood from freely moving laboratory animals and can be coupled for serial2014-05-01 00:00:0010.1002/dta.1646
Dried matrix spotting-An innovative sample preparation tool in bioanalysisAyre, AP;Chaundhari, PS;Shaikh, J;DBSMethod DevelopmentINTERNATIONAL JOURNAL OF PHARMACEUTICAL SCIENCES AND RESEARCH2018https://pdfs.semanticscholar.org/0300/3e93dfc410faa2972b467a56d1d639611856.pdf3597-3607After extraction, samples are subjected to analysis using a suitable analytical technique. Automation: Commercial instruments are available for fully automated online DBS sampling and analysis like ABS2; Instech Solomon, Pennsylvania, USA and Culex; BASi, Indiana, USA2018-01-01 00:00:0010.13040/IJPSR.0975-8232.9(9).3597-07
Effects of corticotropin-releasing factor 1 receptor antagonism on the hypothalamic-pituitary-adrenal axis of rodentsGehlert, DR;Cramer, J;Morin, SM;Hormone ResearchEndocrinologyJ. Pharmacol. Exp. Ther.2012http://jpet.aspetjournals.org/content/341/3/672.short672-680341322402929Corticotropin-releasing factor (CRF) is the major hypothalamic neuropeptide responsible for stimulation of the hypothalamic-pituitary-adrenal axis (HPAA), resulting in the synthesis and release of glucocorticoids from the adrenal cortex. In a recent study, we reported the discovery of the CRF1 receptor antagonist, 3-(4-chloro-2-morpholin-4-yl-thiazol-5-yl)-8-(1-ethylpropyl)-2,6-dimethyl-imidazo[1,2-b]pyridazine (MTIP), which has efficacy in preclinical models of stress-induced alcohol consumption. Because CRF1 is important in HPAA activation, we evaluated the effects of MTIP administration on rodent HPAA function. Initial studies established the MTIP doses required for brain and pituitary CRF1 occupancy and those associated with the inhibition of intracerebroventricular CRF on the HPAA in mice. Then, rat basal plasma corticosterone (CORT) concentrations were measured hourly by radioimmunoassay for 24 h after three daily doses of MTIP or vehicle. In these studies, the early phase of the nocturnal CORT surge was reduced; however, the area under the CORT curve was identical for the 24-h period. In subsequent studies, increases in plasma CORT due to direct pharmacological manipulation of the HPAA axis or by stressors were evaluated after MTIP treatment in mice. MTIP attenuated CORT responses generated by immediate bolus administration of insulin or ethanol; however, MTIP did not affect activation of the HPAA by other stressors and pharmacological agents. Therefore, MTIP can modulate basal HPAA activity during the CORT surge and reduced activation after a select number of stressors but does not produce a lasting suppression of basal CORT. The ability of MTIP to modulate plasma CORT after hyperinsulinemia may provide a surrogate strategy for a target occupancy biomarker.3.615To determine this, we used the BASi Culex API system that allows for administration of the compound via gastric cannulae and blood sampling via implanted jugular cannulae. This method eliminates the need to restrain the animal to accomplish these tasks and the resulting perturbations in stress hormones. For the following studies, all animals were surgically modified and dosed at BASi facilities (West Lafayet2012-06-01 00:00:0010.1124/jpet.111.189753
Efficacy and pharmacokinetic/pharmacodynamic study of 1,1\'-methylenebis{4-[(hydroxyimino)methyl] pyridinium} dimethanesulfonate in guinea pigs and rhesus macaques exposed to cyclosarinHarvilchuck, JA;Hong, SP;Richey, JS;Osheroff, MR;Johnson, JD;PK/PDChemical Weapons ResearchInt. J. Toxicol.2013https://journals.sagepub.com/doi/abs/10.1177/1091581813498425108S-117S324 Suppl23929444Male Hartley guinea pigs and male rhesus macaques were used to determine an efficacious dose of 1,1'-methylenebis{4-[(hydroxyimino)methyl] pyridinium} dimethanesulfonate (MMB4 DMS) that would result in 80% survival, 24 hours following a single exposure to cyclosarin (GF). The pharmacokinetic/pharmacodynamic relationship between acetylcholinesterase activity and MMB4 plasma concentrations relative to survival was evaluated. Guinea pigs and non-human primates (NHPs) were concurrently administered MMB4 DMS (guinea pigs: 0, 10, 30, or 40 mg/kg, intramuscular [IM] and NHPs: 0.1, 1, 5, 10, or 20 mg/kg, IM), atropine, and diazepam following a 3 × median lethal dose (LD50) GF challenge. Clinical observations were evaluated using a quality-of-life (QOL) scoring system. All GF-exposed animals exhibited typical signs of nerve agent poisoning immediately following challenge. In guinea pigs, 24-hour survival was 0%, 50%, 90%, and 90% for 0, 10, 30, and 40 mg/kg MMB4 DMS groups, respectively. In addition, nearly all animals surviving to 24 hours were clinically normal, with many in the 30 and 40 mg/kg MMB4 DMS dose group observed as normal by 4 hours post-challenge. In NHPs, survival was 100% for all treatment groups, with all animals noted as clinically normal by 48 hours. Following treatment with atropine/MMB4 DMS/diazepam, NHPs exhibited dose- and temporal-related decreases in incidence and duration of the clinical signs of toxicity. The QOL scores improved with increasing MMB4 DMS dose in both species. The estimated ED80s were 25.5 mg/kg MMB4 DMS (human equivalent dose [HED] of 5.5 mg/kg) and ≤ 0.1 mg/kg (HED of 0.03 mg/kg) in guinea pigs and NHPs, respectively.1.223Male Hartley guinea pigs and male rhesus macaques were used to determine an efficacious dose of 1,1′-methylenebis{4-[(hydroxyimino)methyl] pyridinium} dimethane... ;Male Hartley guinea pigs (Cavia porcellus), weighing between 250 and 475 g, were purchased from Charles River Laboratories, Inc (Stoneridge, New York). Animals used in the PK/PD phase of the study were surgically implanted with Culex (BASi, West Lafayette, Indiana) catheters in the carotid artery. Animals were single housed in polycarbonate cages on stainless steel racks and were on a 12-hour light/dark cycle2013-08-10 00:00:0010.1177/1091581813498425
Enhancing the Oral Absorption of Kinase Inhibitors Using Lipophilic Salts and Lipid-Based FormulationsWilliams, HD;Ford, L;Han, S;Tangso, KJ;Lim, S;Shackleford, DM;Vodak, DT;Benameur, H;Pouton, CW;Scammells, PJ;Porter, CJH;Pharmacokinetics (PK)?Mol. Pharm.2018https://pubs.acs.org/doi/abs/10.1021/acs.molpharmaceut.8b008585678-5696151230376336The absolute bioavailability of many small molecule kinase inhibitors (smKIs) is low. The reasons for low bioavailability are multifaceted and include constraints due to first pass metabolism and poor absorption. For smKIs where absorption limits oral bioavailability, low aqueous solubility and high lipophilicity, often in combination with high-dose requirements have been implicated in low and variable absorption, food-effects, and absorption-related drug-drug interactions. The current study has evaluated whether preparation of smKIs as lipophilic salts/ionic liquids in combination with coadministration with lipid-based formulations is able to enhance absorption for examples of this compound class. Lipophilic (docusate) salt forms of erlotinib, gefitinib, ceritinib, and cabozantinib (as example smKIs demonstrating low aqueous solubility and high lipophilicity) were prepared and isolated as workable powder solids. In each case, the lipophilic salt exhibited high and significantly enhanced solubility in lipidic excipients (>100 mg/g) when compared to the free base or commercial salt form. Isolation as the lipophilic salt facilitated smKI loading in model lipid-based formulations at high concentration, increased in vitro solubilization at gastric and intestinal pH and in some cases increased oral absorption (∼2-fold for cabozantinib formulations in rats). Application of a lipophilic salt approach can therefore facilitate the use of lipid-based formulations for examples of the smKI compound class where low solubility limits absorption and is a risk factor for increased variability due to food-effects.4.396The rats were kept in Raturn metabolic cages and blood sampling was conducted by a Culex automated blood sampler (ABS) (BASi, West Lafayette, IN, USA). Where animals received drug intravenously, cannulas were also inserted into the jugular vein as described previously.32018-12-03 00:00:0010.1021/acs.molpharmaceut.8b00858
Evaluation of matrix microsampling methods for therapeutic drug candidate quantification in discovery-stage rodent pharmacokinetic studiesSoto, M;Pham, R;Almon, V;Wagner, M;Primack, R;Ponce, M;Meyer, J;James, CA;Salyers, KL;Retter, MW;DBSMethod DevelopmentBioanalysis2014https://www.future-science.com/doi/abs/10.4155/bio.14.1842135-214661625331858AMG 517 or 1-aminobenzotriazole were quantified by LC-MS/MS from low blood/plasma volumes for rat pharmacokinetic (PK) characterization in order to qualify manual/automated dried blood spot (DBS) sampling and plasma separation capillary sampling. In addition, mouse serial automated blood sampling was compared with standard composite sampling. AMG 517 or 1-aminobenzotriazole was administered to rats or mice and multiple microsampling techniques were used to obtain blood or plasma. PK parameters derived from DBS and whole blood-obtained drug concentrations were within 7% for manual DBS and 20% for automated DBS. Plasma PK parameters derived from capillary or standard plasma-obtained drug concentrations differed by 6%. Plasma PK parameters obtained from serial automated blood sampling or manual composite sampling were within 20%. Collectively, these results suggest that the microsampling applications that were investigated are attractive approaches for quantifying drug candidates in low matrix volumes that can be successfully employed within discovery-stage rodent PK studies.2.321AMG 517 or 1-aminobenzotriazole were quantified by LC-MS/MS from low blood/plasma volumes for rat pharmacokinetic (PK) characterization in order to qualify manual/automated dried blood spot (DBS) sampling and plasma separation capillary sampling. In addition, mouse serial automated blood sampling was compared with standard composite sampling. AMG 517 or 1-aminobenzotriazole was administered to rats or mice and multiple microsampling techniques were used to obtain blood or plasma. PK parameters derived from DBS and whole blood-obtained drug concentrations were within 7% for manual DBS and 20% for automated DBS. Plasma PK parameters derived from capillary or standard plasma-obtained drug concentrations differed by 6%. Plasma PK parameters obtained from serial automated blood sampling or manual composite sampling were within 20%. Collectively, these results suggest that the microsampling applications that were investigated are attractive approaches for quantifying drug candidates in low matrix volumes that can be successfully employed within discovery-stage rodent PK studies.2014-08-01 00:00:0010.4155/bio.14.184
Evaluation of Poloxamer as a Slow Release Carrier for MorphineSulimai, NH;Pharmacokinetics (PK)Pain ManagementThesis2017http://search.proquest.com/openview/25dfb275d9de7371611f71f271660d53/1?pq-origsite=gscholar&cbl=18750&diss=yOnce the procedure was finished, Isoflurane was turned off and the rat was tethered to the rat-turn on Culex automated sampling system cage. The rat was allowed 1 day to recover from the surgery and checked for suture breakdown. When the rats were attached to Culex automated sampling system, they received 20 microliters of saline every 12 minutes to keep the catheter patent. The rats were checked for pain and inflammation daily from day of surgical procedure of carotid artery placement until the end of pharmacokinetic study2017-01-01 00:00:00
Evaluation of von Willebrand factor and von Willebrand factor propeptide in models of vascular endothelial cell activation, perturbation, and/or injuryBrott, DA;Katein, A;Thomas, H;Lawton, M;Montgomery, RR;Richardson, RJ;Louden, CS;ADMEVascular InjuryToxicol Pathol2014https://journals.sagepub.com/doi/abs/10.1177/0192623313518664672-68342424499802Pharmacologically, vasoactive agents targeting endothelial and/or smooth muscle cells (SMC) are known to cause acute drug-induced vascular injury (DIVI) and the resulting pathology is due to endothelial cell (EC) perturbation, activation, and/or injury. Alteration in EC structure and/or function may be a critical event in vascular injury and, therefore, evaluation of the circulatory kinetic profile and secretory pattern of EC-specific proteins such as VWF and VWFpp could serve as acute vascular injury biomarkers. In rat and dog models of DIVI, this profile was determined using pharmacologically diverse agents associated with functional stimulation/perturbation (DDAVP), pathological activation (lipopolysaccharide [LPS]/endotoxin), and structural damage (fenoldopam [FD], dopamine [DA], and potassium channel opener (PCO) ZD6169). In rats, FD caused moderate DIVI and time-related increase in plasma VWF levels ∼33% while in control rats VWF increased ∼5%. In dogs, VWF levels transiently increased ∼30% when there was morphologic evidence of DIVI by DA or ZD6169. However, in dogs, VWFpp increased >60-fold (LPS) and >6-fold (DDAVP), respectively. This was in comparison to smaller dynamic 1.38-fold (LPS) and 0.54-fold (DDAVP) increases seen in plasma VWF. Furthermore, DA was associated with a dose-dependent increase in plasma VWFpp. In summary, VWF and VWFpp can discriminate between physiological and pathological perturbation, activation, and injury to ECs. © 2014 by The Author(s).1.382Rats Male Han Wistar rats, 8 to 10 weeks of age (Charles River, Wilmington, MA), were used for all studies. The rats were housed individually in standard polycarbonate cages or in Culex® cages. For intravenous infusion studies (IV), catheters were implanted in the jugular and femoral vein (double catheterization), and the jugular vein catheter was connected to a tethered system and a Harvard pump (KD Scientific, Holliston, MA), while the femoral vein catheter was connected to the Culex® blood collection system (Bioanalytical Systems Inc., West Lafayette, IN). All rats were housed in a temperature-controlled room with 12-hr light/dark cycle and free access to water and food (Purina rodent chow)2014-06-01 00:00:0010.1177/0192623313518664
Exploratory translational modeling approach in drug development to predict human brain pharmacokinetics and pharmacologically relevant clinical dosesKielbasa, W;Stratford, RE;PK and MicrodialysisCNSDrug Metab. Dispos.2012http://dmd.aspetjournals.org/content/40/5/877.short877-88340522287668The central nervous system (CNS) pharmacokinetics (PK) of drugs that have pharmacological targets in the brain are not often understood during drug development, and this gap in knowledge is a limitation in providing a quantitative framework for translating nonclinical pharmacologic data to the clinical patient population. A focus of translational sciences is to improve the efficiency of clinical trial design via a more judicious selection of clinical doses on the basis of nonclinical data. We hypothesize that this can be achieved for CNS-acting drugs based on knowledge of CNS PK and brain target engagement obtained in nonclinical studies. Translating CNS PK models from rat to human can allow for the prediction of human brain PK and the human dose-brain exposure relationship, which can provide insight on the clinical dose(s) having potential brain activity and target engagement. In this study, we explored the potential utility of this translational approach using rat brain microdialysis and PK modeling techniques to predict human brain extracellular fluid PK of atomoxetine and duloxetine. The results show that this translational approach merits consideration as a means to support the clinical development of CNS-mediated drug candidates by enhancing the ability to predict pharmacologically relevant doses in humans in the absence of or in association with other biomarker approaches.3.354In brief, a Culex Automated Pharmacology System (BAS Bioanalytical Systems) was used that enabled timed blood and dialysate collections in awake and freely moving animals. Blood was collected in disodium EDTA tubes and centrifuged to generate plasma within 30 min of collection. At the conclusion of the live phase, animals were killed by CO2 asphyxiation, and whole brain and CSF from the cisterna magnum were collected from each animal. Duloxetine containing blood was removed from brain via carotid arterial perfusion with ice-cold saline before collection. Plasma, dialysate, and CSF samples were stored in a 96-well plate at −70°C before bioanalysis. Whole brains were homogenized in acetonitrile at a ratio of 2 ml/g brain and centrifuged at 3000 rpm for 10 min at an ambient temperature2012-05-01 00:00:0010.1124/dmd.111.043554
Fundamentals, method development, and applications of paper spray ionization mass spectrometryEspy, RD;Pharmacokinetics (PK)Method DevelopmentThesis2014http://search.proquest.com/openview/686ded4f876e7b535baade635595d656/1?pq-origsite=gscholar&cbl=18750&diss=yBlood was collected using a Culex automated blood collection (ABC) system (Bioanalytical Systems Inc., West Lafayette, IN, USA). In this device, the rats were allowed to navigate a 10-inch diameter round cage and remain awake and nonanaesthetized during blood collection. The Culex system drew blood automatically from the central venous catheter, deposited 180 L into an EDTA-coated vial on a carousel, and recycled the remaining blood back into the rat. The blood was then manually aliquotted in preparation for paper spray and LC/MS2014-01-01 00:00:00
Glycoengineered factor IX variants with improved pharmacokinetics and subcutaneous efficacyBrooks, AR;Sim, D;Gritzan, U;Patel, C;Blasko, E;Feldman, RI;Tang, L;Ho, E;Zhao, XY;Apeler, H;Murphy, JE;Pharmacokinetics (PK)HemophiliaJ. Thromb. Haemost.2013https://onlinelibrary.wiley.com/doi/abs/10.1111/jth.123001699-170611923692404The rapid clearance of factor IX (FIX) necessitates frequent intravenous administration to achieve effective prophylaxis for patients with hemophilia B. Subcutaneous administration would be a preferred route of administration but is limited by bioavailability. To improve the pharmacokinetics (PK) and bioavailability of FIX, a screen was performed to identify positions for the introduction of novel glycosylation sites with maximal effect on PK and maintenance of coagulation activity. Two hundred fifty-one variants, each containing one additional N-linked glycosylation site, were screened in vitro, and the PK profiles of selected variants mapping to spatially distinct regions of FIX were evaluated in mice. Optimal variants were combined, and their PK and efficacy were determined in mice with hemophilia B. Variants that mapped to spatially distinct regions of the FIX structure exhibited different degrees of improved PK and enabled selection of optimized sites while minimizing the loss of FIX activity. Combining the most effective N-glycan sites in the same FIX molecule resulted in further improvements in PK. An optimized variant containing three novel N-glycan sites (at amino acids 103, 151, and 228), and the activity enhancing 338A variant had double the specific activity of wild-type FIX, exhibited 4.5-fold reduced clearance and 2.4-fold increased subcutaneous bioavailability, and was efficacious at a fivefold lower mass dose than wild-type FIX after subcutaneous injection in a bleeding model in mice with hemophilia B. Glycoengineering was used to significantly improve the subcutaneous PK and efficacy of FIX and may have advantages for subcutaneous dosing. © 2013 International Society on Thrombosis and Haemostasis.Serial blood samples were collected automatically from the jugular vein via Culex (BASI, Lafayette, IN, USA) into sodium citrate (0.5% final) and plasma was collected after centrifugation at 3300 g. Mice with hemophilia B (Hem B mice) 11 (body weight 18-20 g), were dosed sc ; This ELISA assay detected only human FIX and not rat or mouse FIX2013-09-01 00:00:0010.1111/jth.12300
Green Tea Increases the Concentration of Total Mercury in the Blood of Rats following an Oral Fish Tissue BolusJanle, EM;Freiser, H;Manganais, C;Chen, TY;Craig, BA;Santerre, CR;ADMENutritional SupplementsBiomed Res Int2015https://www.hindawi.com/journals/bmri/aa/320936/320936201526301246Fish has many health benefits but is also the most common source of methylmercury. The bioavailability of methylmercury in fish may be affected by other meal components. In this study, the effect of green tea on the bioavailability of methylmercury from an oral bolus of fish muscle tissue was studied in rats and compared to a water treated control group and a group treated with meso-2,3-dimercaptosuccinic acid (DMSA), a compound used medically to chelate mercury. Rats were given a single oral dose of fish tissue via gavage and one of the treatments. Rats were given access to food for 3 h at 12 h intervals. They were dosed with each of the treatments with each meal. Blood samples were collected for 95 hours. Green tea significantly increased the concentration of total mercury in blood relative to the control, whereas DMSA significantly decreased it. In addition, feeding caused a slight increase in blood mercury for several meals following the initial dose.2.197Rats were surgically implanted with a femoral vein catheter under isoflurane anesthesia. They were placed in a Culex automated in vivo sampling system (Bioanalytical Systems, West Lafayette, IN). Rats were allowed to recover from surgery for 48 hours. To maintain patency the Culex injects 15 μL of dilute heparinized saline (20 U/mL) every 10 minutes2015-08-02 00:00:0010.1155/2015/320936
Hepatocyte clearance and pharmacokinetics of recombinant factor IX glycosylation variantsBlasko, E;Brooks, AR;Ho, E;Wu, JM;Zhao, XY;Subramanyam, B;ADMEDrug MetabolismBiochem. Biophys. Res. Commun.2013https://www.sciencedirect.com/science/article/pii/S0006291X13014617485-489440424036269Addition of N-linked glycosylation sites has been shown to increase serum half-life and decrease clearance for proteins such as recombinant erythropoietin (EPO). However, factor IX (FIX) variants with additional N-linked glycans ("HG" variants) that were expressed in HKB11 cells showed increased clearance in rat in vivo pharmacokinetic studies relative to FIX variants with no additional glycans. Variants with multiple additional glycans were the most rapidly cleared. A rat hepatocyte clearance assay was developed to measure intrinsic clearance of these FIX variants in vitro. The rank order of clearance of the variants was the same both in vivo and in the in vitro hepatocyte assay. In the in vitro assay, heparin, galactose, and asialo-orosomucoid inhibited clearance of a FIX HG variant by hepatocytes, and asialo-FIX was rapidly cleared, suggesting roles for the asialoglycoprotein receptor (ASGPR) and cell surface proteoglycans in FIX clearance. Thus the in vitro hepatocyte intrinsic clearance assay is both useful and predictive for identifying rapidly cleared recombinant proteins and for helping to identify receptors involved in clearance of proteins by the liver. Copyright © 2013 Elsevier Inc. All rights reserved.2.705Sprague-Dawley rats (Harlan Laboratories, Indianapolis, IN), approximately 10 weeks old, were dosed intravenously at 0.70 mg/kg (bolus injection) with FIX variants (n = 4). Blood samples were collected automatically through the jugular vein via Culex® (BASI, Lafayette, IN) at ;cell-expressed glycosylation variants (HG9, HG3/9/10, HG3/5/9). Blood samples were drawn at various time points using Culex™ (BASI, Lafayette, IN Primary rat hepatocytes, 1 million viable cells/ml, were incubated with 25 ng/ml of various FIX variants with mixing at 37 °C in a 2013-11-01 00:00:0010.1016/j.bbrc.2013.09.001
Historical Perspective on Blood and PhlebotomyKissinger, PT;Pharmacokinetics (PK)Data Reviewtechnical note2012https://phlebot.com/assets/pdf/Tech%20Note%20-%20History.pdf1-5originally developed by a research team led by Candice B. Kissinger at Bioanalytical Systems Inc. in 2000 (7). The purpose of the device (Culex®) was to facilitate in vivo extended to larger rodents (guinea pigs) and smaller (mice), and then larger animals (dog, monkey, pig)2012-01-01 00:00:00
How a rodent is dosed and sampled is equally important to how samples are analyzed: Automated Dosing, Sampling and LC-MS/MS with ion traps and tripleZhu, Y;Kissinger, BC;Kissinger, TP;Bromet, N;PK/PDMethod DevelopmentResearchgate2005https://www.researchgate.net/profile/Peter_Kissinger/publication/268324652_How_a_rodent_is_dosed_and_sampled_is_equally_important_to_how_the_samples_are_analyzed_automated_dosing_sampling_and_LCMSMS_with_ion_traps_and_triple_quads/links/09e4150feb225d8375000000.pdf1-13done and costs virtually nothing once an animal is instrumented for collecting other data (www.culex.net) (This paper was published in Current Separations, 2005, 21(2):37, an internal publication of Bioanalytical Systems, Inc.) The history of automated blood sampling at BASi2005-01-01 00:00:00
Identification and quantification of the oxidized protein profile in the Zucker diabetic fatty rat supplemented with green teaMyracle, AD;ADMENutritional SupplementsThesis2010http://search.proquest.com/openview/7b9d2f803a67e7e639bf96d59cc425c6/1?pq-origsite=gscholar&cbl=18750&diss=yPage 1. Graduate School ETD Form 9 (Revised 12/07) PURDUE UNIVERSITY GRADUATE SCHOOL Thesis/Dissertation Acceptance This is to certify that the thesis/dissertation prepared By Entitled For the degree of Is approved by the final examining committee: Chair ;Prior to waking, the animals were placed in the Culex (BAS, Inc., West Lafayette, IN) where the UF probe line was fed through the cage apparatus and exteriorized to a small peristaltic pump set on a speed of 4 for overnight collection of interstitial fluid. The line was attached to a needle hub which was inserted into a vacutainer® (BD, Franklin Lakes, NJ) and placed on ice2010-01-01 00:00:00
Identification of Three Novel Ring Expansion Metabolites of KAE609, a New Spiroindolone Agent for the Treatment of Malaria, in Rats, Dogs, and HumansHuskey, SE;Zhu, CQ;Lin, MM;Forseth, RR;Gu, H;Simon, O;Eggimann, FK;Kittelmann, M;Luneau, A;Vargas, A;Li, H;Wang, L;Einolf, HJ;Zhang, J;Favara, S;He, H;Mangold, JB;Pharmacokinetics (PK)AntimalarialDrug Metab. Dispos.2016http://dmd.aspetjournals.org/content/44/5/653.short653-66444526921386KAE609 [(1'R,3'S)-5,7'-dichloro-6'-fluoro-3'-methyl-2',3',4',9'-tetrahydrospiro[indoline-3,1'-pyridol[3,4-b]indol]-2-one] is a potent, fast-acting, schizonticidal agent being developed for the treatment of malaria. After oral dosing of KAE609 to rats and dogs, the major radioactive component in plasma was KAE609. An oxidative metabolite, M18, was the prominent metabolite in rat and dog plasma. KAE609 was well absorbed and extensively metabolized such that low levels of parent compound (≤11% of the dose) were detected in feces. The elimination of KAE609 and metabolites was primarily mediated via biliary pathways (≥93% of the dose) in the feces of rats and dogs. M37 and M23 were the major metabolites in rat and dog feces, respectively. Among the prominent metabolites of KAE609, the isobaric chemical species, M37, was observed, suggesting the involvement of an isomerization or rearrangement during biotransformation. Subsequent structural elucidation of M37 revealed that KAE609, a spiroindolone, undergoes an unusual C-C bond cleavage, followed by a 1,2-acyl shift to form a ring expansion metabolite M37. The in vitro metabolism of KAE609 in hepatocytes was investigated to understand this novel biotransformation. The metabolism of KAE609 was qualitatively similar across the species studied; thus, further investigation was conducted using human recombinant cytochrome P450 enzymes. The ring expansion reaction was found to be primarily catalyzed by cytochrome P450 (CYP) 3A4 yielding M37. M37 was subsequently oxidized to M18 by CYP3A4 and hydroxylated to M23 primarily by CYP1A2. Interestingly, M37 was colorless, whereas M18 and M23 showed orange yellow color. The source of the color of M18 and M23 was attributed to their extended conjugated system of double bonds in the structures. Copyright © 2016 by The American Society for Pharmacology and Experimental Therapeutics.3.354Male Wistar Hannover rats (approximately 227–310 g, n = 12) were purchased from Harlan Laboratories (South Easton, MA). Catheters were surgically implanted into the carotid artery and/or jugular vein of rats by the vendor (only one catheter was implanted into carotid artery for blood collection from rats receiving the oral dose). All rats were housed individually in Culex metabolism cages (Culex Autosampler; BAS, Indianapolis, IN) in a temperature- and humidity-controlled room (22 ± 2°C; 50% ± 15%) with free access to food and water (food was withheld until 4 hours postdose)2016-05-01 00:00:0010.1124/dmd.115.069112
Immunomodulatory Properties, Mechanisms, and Therapeutic Potential of Macrocyclic Theta DefensinsSchaal, JB;PK/PDImmune ResponseThesis2016http://search.proquest.com/openview/5fb2d372d40125d8d0c7f3a032b6e1b0/1?pq-origsite=gscholar&cbl=18750&diss=yhoused (Culex NxT Automated In Vivo Sampling System; BASi), with ad lib access to water and standard chow and 12 hour light/dark cycle. RTD-1 dissolved in saline was injected subcutaneously at 5 mg/kg to 4 animals. Approximately 200 μL of blood was collected prior to peptide administration and 0.25, 0.5, 1, 2, 4, 6, 8, 12, 18, 24, 30, 36, 48, 60, and 72 h after RTD1 injection. Blood was collected into pre-chilled (4° C) vials containing K3EDTA and plasma2016-01-01 00:00:00
Impacts on Sirtuin Function and Bioavailability of the Dietary Bioactive Compound DihydrocoumarinJacobi, JL;Yang, B;Li, X;Menze, AK;Laurentz, SM;Janle, EM;Ferruzzi, MG;McCabe, GP;Chapple, C;Kirchmaier, AL;ADMENutritional SupplementsPLoS ONE2016https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0149207e014920711226882112The plant secondary metabolite and common food additive dihydrocoumarin (DHC) is an inhibitor of the Sirtuin family of NAD+-dependent deacetylases. Sirtuins are key regulators of epigenetic processes that maintain silent chromatin in yeast and have been linked to gene expression, metabolism, apoptosis, tumorogenesis and age-related processes in multiple organisms, including humans. Here we report that exposure to the polyphenol DHC led to defects in several Sirtuin-regulated processes in budding yeast including the establishment and maintenance of Sir2p-dependent silencing by causing disassembly of silent chromatin, Hst1p-dependent repression of meiotic-specific genes during the mitotic cell cycle. As both transient and prolonged exposure to environmental and dietary factors have the potential to lead to heritable alterations in epigenetic states and to modulate additional Sirtuin-dependent phenotypes, we examined the bioavailability and digestive stability of DHC using an in vivo rat model and in vitro digestive simulator. Our analyses revealed that DHC was unstable during digestion and could be converted to melilotic acid (MA), which also caused epigenetic defects, albeit less efficiently. Upon ingestion, DHC was observed primarily in intestinal tissues, but did not accumulate over time and was readily cleared from the animals. MA displayed a wider tissue distribution and, in contrast to DHC, was also detected in the blood plasma, interstitial fluid, and urine, implying that the conversion of DHC to the less bioactive compound, MA, occurred efficiently in vivo.2.776Automatic blood and interstitial fluid draws was performed using a Culex automated sampling system (BASi, West Lafayette, IN), which infused 10 μl heparinized saline every 10 min to maintain catheter patency. Rats were given 100 mg/kg body mass dose of DHC in a 1 ml emulsion of DHC, water and Tween 20 by gavage (see in vitro digestion methods above). Food was restricted 8 hr prior to gavage and offered 2 hr post-gavage. The CulexTM was used to automatically draw 250 μl of blood from the femoral catheter into heparinized tubes2016-02-16 00:00:0010.1371/journal.pone.0149207
Improved extrapolation of hepatobiliary clearance from in vitro sandwich cultured rat hepatocytes through absolute quantification of hepatobiliary transportersLi, N;Singh, P;Mandrell, KM;Lai, Y;ADME (Bile)?Mol. Pharm.2010https://pubs.acs.org/doi/abs/10.1021/mp9001574630-6417320438085Previously we have reported that hepatobiliary transporter expressions in sandwich cultured hepatocytes (SCH) are altered 2- to 5-fold. This change could limit the model's predictive power for in vivo biliary clearance. The present study was designed to better establish in vitro to in vivo correlation (IVIVC) of biliary clearance. Eleven compounds representing the substrates of Mrp2/Abcc2, Bcrp/Abcg2 and Bsep/Abcb11 were tested in the sandwich cultured rat hepatocyte (SCRH) model. Simultaneously, the absolute difference of hepatobiliary transporters between rat livers and SCRH at day 5 post culture was determined by LC-MS/MS. This difference was integrated into the well-stirred hepatic prediction model. A correction factor named "g_factor" was mathematically defined to reflect the difference in hepatobiliary transporter expressions between the SCRH model and in vivo models, as well as the contribution of multiple transporters. When the g_factor correction was applied, the in vivo biliary clearance prediction was significantly improved. In addition, for those compounds which are poorly permeable and/or undergo transporter-dependent active uptake, the known intracellular concentrations of substrates were used to estimate intrinsic bile clearance. This led to further improvement in the prediction of in vivo bile secretion. While the rate-limiting processes of uptake transporters in the SCRH model remain to be further determined, we showed that integration of the absolute difference of hepatobiliary transporter proteins and transport contributions could improve the predictability of SCRH model. This integration is fundamental for increased confidence in the IVIVC of human biliary clearance.4.396The pooled rat plasma was spiked with 2 μL of 2.5 mM stock solutions in The rats were surgically implanted with BASi vascular catheters (West Lafayette, IN) in carotid artery Animals were acclimated in Culex cages (Bioanalytical Services, Inc., West Lafayette, IN) overnight prior2010-06-07 00:00:0010.1021/mp9001574
In the LC LiteratureMora, MI;Marioli, JM;Chrom, JL;PK and MicrodialysisMethod DevelopmentCurrent Separations2002http://www.currentseparations.com/issues/19-4/19-4g.pdf1-6(www.culex.net) New brain microdialysis probes with intracerebral cannulae have been developed by Bioanalytical Systems, Inc The MBR line of probes expands the offerings of microdialysis probes available from BAS for studies in brain, dermis, bile, blood vessels and other2002-01-01 00:00:00
In the MD LiteratureMicrodialysis, H;Microdialysis, CU;ADMENutritional SupplementsBiotechnology2002https://www.researchgate.net/profile/Jan_Kehr/publication/237569236_Monitoring_Molecules_in_Neuroscience_MicaGenix_A_New_BASi_Partner_through_PiCRA/links/0c96052998bad39b57000000.pdf1-5with Multi-Channel Electrochemical Detection for the Determination of Daidzin in Rat Blood Sampled New Rodent Cage for Culex Blood Samplers, Rodent Workstations and Microdialysis Applications A new size of BASi Chads is now available for standard Eppendorf® tubes2002-01-01 00:00:00
In vitro and in vivo evaluation of a novel water-soluble N-glycyl prodrug (N-GLY-CBZ) of carbamazepineHemenway, JN;Stella, VJ;Pharmacokinetics (PK)AnticonvulsantJ Pharm Sci2010https://www.sciencedirect.com/science/article/pii/S00223549153241264565-4575991120845455The synthesis and characterization of N-glycyl-carbamazepine (N-Gly-CBZ), an N-acyl urea derivative of carbamazepine (CBZ) designed to act as a prodrug and convert to CBZ and glycine in vivo by enzymatic cleavage of the glycyl-urea bond was recently reported. The rate and extent of conversion of N-Gly-CBZ to CBZ in a whole animal model is reported here along with supporting in vitro data. Pharmacokinetic parameters were determined for N-Gly-CBZ and CBZ following IV and oral administration of N-Gly-CBZ and CBZ control to rats using a crossover design. The in vivo elimination of N-Gly-CBZ following IV administration in rats was biphasic in nature with a t(1/2) of about 1.1 min, which was very similar to the t(1/2) for appearance of CBZ. The mean value for the relative AUC ratio for CBZ from N-Gly-CBZ and CBZ from a cyclodextrin solution showed that N-Gly-CBZ delivered a (± SD) 98 ± 16% (± SD) equivalent dose of CBZ in six rats. The results of the IV dosing pharmacokinetics investigation were consistent with N-Gly-CBZ acting as a prodrug with rapid and complete conversion to CBZ in vivo. The overall absolute oral bioavailability of CBZ from N-Gly-CBZ was determined to be 41 ± 14% in three rats. The relative oral bioavailability of CBZ from N-Gly-CBZ compared to an oral CBZ control was 1.72 ± 0.54. That is, the prodrug, N-Gly-CBZ, demonstrated superior oral bioavailability of CBZ over the CBZ control, which was likely due to its greater aqueous solubility. © 2010 Wiley-Liss, Inc. and the American Pharmacists Association3.197neck where the cannulae were externalized and placed within a swivel device allowing the rat free mobility and access to water. Upon completion of surgery, the rats were tethered in a metabolism cage of the Culex Automated Blood Sampler (ABS), Bioanalytical Systems, Inc 2010-11-01 00:00:0010.1002/jps.22171
In vitro-in vivo correlation and translation to the clinical outcome for CJ-13,610, a novel inhibitor of 5-lipoxygenaseMatthew Hutzler, J;Linder, CD;Melton, RJ;Vincent, J;Daniels, JS;Translational PharmacologyDrug MetabolismDrug Metab. Dispos.2010http://dmd.aspetjournals.org/content/38/7/1113.short1113-112138720375180The metabolism of the 5-lipoxygenase inhibitor, 4-(3-(4-(2-methyl-1H-imidazol-1-yl)phenylthio)phenyl)-tetrahydro-2H-pyran-4-carboxamide (CJ-13,610), was investigated in liver microsomes from human and preclinical species in an effort to compare metabolite profiles and evaluate the in vitro-in vivo correlation for metabolic clearance. Overall, the metabolite profile of CJ-13,610 was comparable across the species tested with multiple oxidative metabolites observed, including sulfoxidation. The sulfoxidation kinetics characterized in rat, dog, and human liver microsomes (HLM) indicated a low apparent Michaelis-Menten constant (K(m, app)) of 4 to 5 microM. Results from cDNA-expressed cytochrome P450 (P450) studies indicated that the metabolism in HLM was primarily mediated by CYP3A4 and 3A5. A subsequent in vitro study using ketoconazole as an inhibitor of CJ-13,610 sulfoxidation corroborated the CYP3A4/5-mediated pathway (IC(50) = 7 nM). Assessment of multiple methods for predicting the human pharmacokinetic profile observed with CJ-13,610 after a 30-mg single oral dose indicated that clearance scaled from human liver microsomes yielded a better prediction when coupled with a Vd(ss) term that was scaled from dog [area under the concentration-time curve (AUC) and half-life within 1.3-fold of actual] versus a Vd(ss) term obtained from rat. Single-species allometric scaling of clearance and Vd(ss) from dog pharmacokinetic studies was equally predictive, whereas scaling from rat resulted in underpredictions of both AUC and maximal concentration (C(max)). Results from these studies support the strategy of predicting human pharmacokinetics using human liver microsomal intrinsic clearance data. More importantly, results from the present investigation enabled the selection of alternative drug candidates from the chemical series via in vitro screening, while subsequently eliminating costly routine preclinical in vivo studies.3.354Male Sprague-Dawley rats were purchased from Charles River Laboratories, Inc. (Wilmington, MA) and acclimated to their surroundings for approximately 1 week with food and water provided ad libitum. On the day before the study, animals were anesthetized with isoflurane (to effect) and then implanted with BASi vascular catheters in the carotid artery and jugular vein. Animals were acclimated in Culex cages overnight before dosing. Patency of the carotid artery catheter was maintained using the “tend” function of a Culex ABS. Three rats were dosed intravenously with a 10% ethanol-56% PEG-400-34% phosphate-buffered saline (pH 7.4) at 0.5 mg/kg via the jugular vein catheter at a dose volume of 0.5 ml/kg. Blood collections were performed by the Culex ABS predose and at 2, 5, 15, and 30 min and 1, 2, 4, 6, 8, 12, 18, and 24 h postdose; samples were collected into chilled heparinized tubes and centrifuged for 10 min at 3000 rpm, and the resulting plasma was aliquoted to 96-well plates for LC-MS/MS analysis. Male beagle dogs (approximately 1–3 years of age and weighing between 9 and 12 kg at study initiation) were obtained from Marshall Farms (North Rose, NY)2010-07-01 00:00:0010.1124/dmd.110.032706
In vivo metabolic tracking of 14C-radiolabelled isoflavones in kudzu (Pueraria lobata) and red clover (Trifolium pratense) extractsMun, JG;Grannan, MD;Lachcik, PJ;Reppert, A;Yousef, GG;Rogers, RB;Janle, EM;Weaver, CM;Lila, MA;ADMENutritional SupplementsBr. J. Nutr.2009https://www.cambridge.org/core/journals/british-journal-of-nutrition/article/in-vivo-metabolic-tracking-of-14cradiolabelled-isoflavones-in-kudzu-pueraria-lobata-and-red-clover-trifolium-pratense-extracts/D812AE5CE62F8976DC01CC0D66BED9061523-15301021019586574Absorption, distribution and elimination of 14C-labelled isoflavone-containing extracts from kudzu (Pueraria lobata) root culture and red clover (Trifolium pratense) cell culture were investigated in an in vivo rat model. The predominant isoflavones in the kudzu extract were the glycosides puerarin, daidzin and malonyl daidzin, while in the red clover extract, the major isoflavones were formononetin and its derivatives, genistein and biochanin A, with radioactivities of 3.770 and 7.256 MBq/g, respectively. Male Sprague-Dawley rats, implanted with a jugular catheter and a subcutaneous ultrafiltrate probe, were orally administered with 14C-labelled isoflavone extracts from either kudzu or clover cell cultures. Serum, interstitial fluid (ISF), urine and faeces were collected using a Culex Automated Blood Collection System for 24 h. Analysis of bone tissues revealed that radiolabel accumulated in the femur, tibia and vertebrae at 0.04, 0.03 and 0.01 % of the administered dose, respectively, in both kudzu and red clover treatments. The liver accumulated the greatest concentration of radiolabel among the tissues tested, at 1.99 and 1.54 % of the administered kudzu and red clover extracts, respectively. Serum and ISF analysis showed that both extracts were rapidly absorbed, distributed in various tissues, and largely eliminated in the urine and faeces. Urine and faeces contained 8.53 and 9.06 % of the kudzu dose, respectively, and 3.60 and 5.64 % of the red clover dose, respectively. Serum pharmacokinetics suggest that extracts from kudzu may undergo enterohepatic circulation.3.319Absorption, distribution and elimination of 14C-labelled isoflavone-containing extracts from kudzu (Pueraria lobata) root culture and red clover (Trifolium pratense) cell culture were investigated in an in vivo rat model. The predominant isoflavones in the kudzu extract were the glycosides puerarin, daidzin and malonyl daidzin, while in the red clover extract, the major isoflavones were formononetin and its derivatives, genistein and biochanin A, with radioactivities of 3.770 and 7.256 MBq/g, respectively. Male Sprague-Dawley rats, implanted with a jugular catheter and a subcutaneous ultrafiltrate probe, were orally administered with 14C-labelled isoflavone extracts from either kudzu or clover cell cultures. Serum, interstitial fluid (ISF), urine and faeces were collected using a Culex Automated Blood Collection System for 24 h. Analysis of bone tissues revealed that radiolabel accumulated in the femur, tibia and vertebrae at 0.04, 0.03 and 0.01 % of the administered dose, respectively, in both kudzu and red clover treatments. The liver accumulated the greatest concentration of radiolabel among the tissues tested, at 1.99 and 1.54 % of the administered kudzu and red clover extracts, respectively. Serum and ISF analysis showed that both extracts were rapidly absorbed, distributed in various tissues, and largely eliminated in the urine and faeces. Urine and faeces contained 8.53 and 9.06 % of the kudzu dose, respectively, and 3.60 and 5.64 % of the red clover dose, respectively. Serum pharmacokinetics suggest that extracts from kudzu may undergo enterohepatic circulation2009-11-01 00:00:0010.1017/S000711450999047X
INFLUENCE OF FLUVOXAMINE ON CARVEDILOL’S PHARMACOKINETICS IN RATSABRUDAN, M;Pharmacokinetics (PK)Drug-drug InteractionFARMACIA2019http://www.revistafarmacia.ro/201904/2019-04-art-09-Abrudan_Muntean_Neag_616-620.pdf616-6206741.527The connection of the animal to BASi Culex ABC® needs the cannulation on the left femoral vein of the rat The venous cannula was used for blood sampling. The cannulation procedure was realized before connecting the rat to BASi Culex ABC®2019-07-10 00:00:0010.31925/farmacia.2019.4.9
Inhibitory Effect of Citalopram on the Pharmacokinetics of Carvedilol in Rats and in vitro ModelsAbrudan, MB;Muntean, DM;Popa, DS;Gheldiu, AM;Neag, MA;Vlase, L;Pharmacokinetics (PK)Drug-drug InteractionPharmacology2017https://www.karger.com/Article/Abstract/480090301-3071005-628848215The aim of this study was to investigate the drug-drug interaction between carvedilol and citalopram based on carvedilol metabolism in vitro and his pharmacokinetics (PKs) in vivo after the oral administration of the single drug and both drugs, and reveal citalopram effects on the PKs of carvedilol. Each rat was cannulated on the femoral vein, prior to being connected to BASi Culex ABC®. Carvedilol was orally administrated in rats (3.57 mg/kg body weight [b.w.]) in the absence of citalopram or after a pre-treatment with multiple oral doses of citalopram (1.42 mg/kg b.w.). Plasma concentrations of carvedilol were determined using high-performance liquid chromatography-MS at the designated time points after drug administration, and the main PK parameters were calculated by noncompartmental analysis. In addition, effects of citalopram on the metabolic rate of carvedilol were investigated using rat-pooled liver microsome incubation systems. During co-administration, significant increases of the area under the plasma concentration-time curve as well as of the peak plasma concentration were observed. The rat-pooled liver microsome incubation experiment indicated that citalopram could decrease the metabolic rate of carvedilol. Citalopram co-administration led to a significant alteration of carvedilol's PK profile in rats; it also demonstrated, in vitro, these effects could be explained by the existence of a drug-drug interaction mediated by CYP2D6 inhibition. © 2017 S. Karger AG, Basel.1.615drugs, and reveal citalopram effects on the PKs of carvedilol. Methods: Each rat was cannulated on the femoral vein, prior to being connected to BASi Culex ABC®. Carvedilol was orally administrated in rats (3.57 mg/kg body2017-08-19 00:00:0010.1159/000480090
Integration of cardiac energetics, function and histology from isolated rat hearts perfused with doxorubicin and doxorubicin-ol; a model for use in drug safety evaluationsHenderson, KA;Borders, RB;Ross, JB;Abdulalil, A;Gibbs, S;Skowronek, AJ;Knostman, K;Bailey, J;Smith, J;Vinci, T;Wood, B;Knopp, MV;Roche, BM;Pharmacokinetics (PK)OncologyJ Pharmacol Toxicol Methods2018https://www.sciencedirect.com/science/article/pii/S105687191830589654-6394Pt 230195582The isolated rat heart (Langendorff) assay combined with NMR spectroscopy and histology were used to elucidate functional, metabolic, and histological signs of cardiotoxicity resulting from acute exposure to clinically relevant concentrations of doxorubicin and its metabolite dox-ol. Doxorubicin blood concentrations and pharmacokinetic parameters were assessed following a clinically relevant dose of 2 mg/kg in order to select concentrations for isolated heart perfusions. Isolated rat hearts were exposed to 1 or 10 μM of doxorubicin or 0.3 μM dox-ol for at least 60 min using the Langendorff perfusion method. Effects on heart function were monitored using ECGs, left ventricular contraction parameters, and microscopic histology. Cardiac energetics (PCr, ATP, and Pi) were evaluated before, during, and after exposure to doxorubicin/dox-ol in perfused hearts using NMR spectroscopy. Cardiac effects were evident following clinically relevant concentrations of doxorubicin and dox-ol in isolated rat hearts demonstrated by altered heart function, energetic reserve, and microscopic lesions. A cardiac stress test utilizing isoproterenol resulted in enhanced functional response and reductions in PCr in doxorubicin versus vehicle treated hearts indicating possible alterations in the isoproterenol mediated pathway. Dox-ol treated hearts were similar to control with regard to function, but exhibited histologic findings. The use of combined Langendorff/NMR/histology methodologies allowed for comparison of multiple indices of cardiac function at one time in which cardiac effects were evident in multiple parameters. SHORT ABSTRACT: The isolated rat heart assay combined with NMR spectroscopy and histology was used to elucidate functional, metabolic, and histological signs of cardiotoxicity resulting from acute exposure to clinically relevant concentrations of doxorubicin and its metabolite dox-ol. Heart function was altered and microscopic signs of toxicity were evident with dox and dox-ol exposures. The use of combined Langendorff/NMR/histology assays allowed for comparison of multiple indices of cardiac function at one time in which cardiac effects were evident in multiple parameters. Copyright © 2018 Elsevier Inc. All rights reserved.2.679The specific objectives were to develop a toxicokinetic profile for doxorubicin in the rat and to establish from the PK data the was collected at approximately 2, 5, 8, 11, 15, 30, 45, and 60 min post dose using a Culex automated blood collection system (Basi, West Lafayette 2018-09-06 00:00:0010.1016/j.vascn.2018.08.004
Involvement of intestinal uptake transporters in the absorption of azithromycin and clarithromycin in the ratGarver, E;Hugger, ED;Shearn, SP;Rao, A;Dawson, PA;Davis, CB;Han, C;Pharmacokinetics (PK)AntibioticsDrug Metab. Dispos.2008http://dmd.aspetjournals.org/content/36/12/2492.short2492-2498361218755851Macrolide antibiotics azithromycin (AZI) and clarithromycin (CLARI) are large molecular weight compounds and are substrates for apically polarized efflux transporters such as P-glycoprotein, which can potentially restrict intestinal absorption. However, despite these undesired physicochemical and biopharmaceutical properties, AZI and CLARI exhibit moderate to excellent p.o. bioavailability in preclinical species and humans. Intestinal uptake transporters, such as organic anion transporting polypeptides (OATPs), can facilitate the uptake of drugs that are substrates and hence increase p.o. absorption. The present study was designed to determine whether the intestinal Oatps are involved in absorption of these macrolides. AZI or CLARI was dosed p.o. to Sprague-Dawley rats after p.o. administration with vehicle or rifamycin SV (RIF), an OATP inhibitor. The p.o. exposures of AZI and CLARI were reduced 65 and 45%, respectively, when coadministered with an optimized RIF regimen. The p.o. RIF had no affect on the total blood clearance of these macrolides and most likely did not cause induction of metabolizing enzymes and/or transporters. Therefore, the results suggest that inhibition of an RIF-sensitive uptake transporter such as Oatp along the rat gastrointestinal tract was responsible for reduced p.o. exposure of AZI and CLARI. In addition, AZI and CLARI caused inhibition of taurocholate uptake in rat Oatp1a5-transfected Madin-Darby canine kidney cell monolayers. The in vitro and in vivo results suggest that the intestinal Oatps are involved in the p.o. absorption of AZI and CLARI in the rat.3.354Male Sprague-Dawley rats (325– 425 g) were purchased from Charles River Laboratories (Raleigh, NC) and were cannulated with Tygon tubing (Norton Performance Plastics, Akron, OH) femoral artery and vein catheters at least 2 days before study initiation. Animals were tethered, placed in an automated blood sampling system (Culex, Bioanalytical Systems Incorporated, West Lafayette, IN), and acclimated for a minimum of 24 h before dose administration. Catheter patency was maintained with regular flushing intervals with 20 IU/ml heparin derived from porcine intestinal mucosa (Abraxis Pharmaceutical Products, Schaumburg, IL) throughout the study and maintained under standard conditions with a 12-h light/dark cycle. All the animal protocols were approved by GlaxoSmithKline Animal Care and Use Committee and were conducted in accordance with International Animal Care and Use Committee policy2008-12-01 00:00:0010.1124/dmd.108.022285
Isotopic labeling of isoflavones from plant cell and organ culture for metabolic tracking in animal modelsMun, JG;PK and MicrodialysisNutritional SupplementsThesis2010http://www.ideals.illinois.edu/handle/2142/16006Male Sprague-Dawley rats, implanted with a jugular catheter and a subcutaneous ultrafiltrate probe, were gavaged with 14C-labeled isoflavone extracts from either kudzu or clover cell cultures, and serum, interstitial fluid (ISF), urine and feces were collected using a Culex Automated Blood Collection System for 24 h. Analysis of bone tissues revealed that radiolabel accumulated in the femur, tibia, and vertebrae at 0.04%, 0.03%, and 0.01% of the administered dose respectively in both kudzu and red clover treatments2010-01-01 00:00:00
Linking internal dosimetries of the propyl metabolic series in rats and humans using physiologically based pharmacokinetic (PBPK) modelingSmith, JN;Tyrrell, KJ;Smith, JP;Weitz, KK;Faber, W;Pharmacokinetics (PK)Environmental ToxicologyRegul. Toxicol. Pharmacol.2020https://www.sciencedirect.com/science/article/pii/S027323001930271510450711031669189The metabolic series approach has successfully linked internal dosimetries of metabolically related compounds reducing cost and time for chemical risk assessments. Here, we developed a physiologically based pharmacokinetic (PBPK) model in rats and humans for the propyl metabolic series including propyl acetate, 1-propanol, propionaldehyde, and propionic acid. Manufacturers use these compounds as organic solvents and intermediates during chemical synthesis. Public exposures can occur through using consumer products containing propyl compounds like cosmetics, aerosol sprays, or foods, and occupational exposures can occur at manufacturing facilities. To develop the PBPK model, we measured in vitro metabolism of propyl acetate in blood and liver S9 fractions. We measured concentrations of propyl compounds in blood following intravenous (iv) infusion of 13C-propanol or 13C-propionic acid and closed chamber inhalation exposures to propyl acetate or propanol in rats. Using these studies and other published data, we modified an existing PBPK model for the butyl metabolic series to simulate time course concentrations of propyl compounds in rats and humans. Consistent with measured in vitro and in vivo data, the optimized propyl series model predicts rapid clearance of propyl acetate, higher concentrations of propanol in blood from propyl acetate inhalation compared to propanol inhalation in rats but not in humans, and low concentrations of propionic acid in blood from exposures to propyl acetate or propanol. Regulators can use this model as a tool for propyl compound risk assessment by linking internal dosimetries under various exposure scenarios. Copyright © 2019 Elsevier Inc. All rights reserved.2.996Isotopic propionic acid (~80 μM final concentration) was spiked into phosphate buffer or rat plasma diluted in phosphate buffer (10%) animals selected for dosing were housed in the Raturn Housing System, which is part of the Culex/Empis (BASi, West Lafayette, IN)2020-02-01 00:00:0010.1016/j.yrtph.2019.104507
LLY-2707, a novel nonsteroidal glucocorticoid antagonist that reduces atypical antipsychotic-associated weight gain in ratsSindelar, DK;Carson, MW;Morin, M;Shaw, J;Barr, RJ;Need, A;Alexander-Chacko, J;Coghlan, M;Gehlert, DR;Pharmacokinetics (PK)AntipsychoticJ. Pharmacol. Exp. Ther.2014http://jpet.aspetjournals.org/content/348/1/192.short192-201348124163440Weight gain and diabetes have been reported during treatment with atypical antipsychotic drugs (AAPDs). Patients treated with the glucocorticoid receptor antagonist (GRA) and the progesterone receptor antagonist (PRA) mifepristone [estra-4,9-dien-3-one, 11-[4-(dimethylamino)phenyl]-17-hydroxy-17-(1-propynyl)-(11β,17β)-(9CI)] experienced significant reduction in the weight gain observed when patients were treated with olanzapine or risperidone. To understand the pharmacology responsible for this finding, we discovered LLY-2707 [N-(5-(tert-butyl)-3-(2-fluoro-5-methylpyridin-4-yl)-2-methyl-1H-indol-7-yl)methanesulfonamide], a novel and selective GRA, and evaluated its utility in preclinical models of AAPD-associated weight gain and diabetes. In vitro, LLY-2707 was a highly selective and potent GRA. GR occupancy in vivo was assessed using ex vivo binding where LLY-2707 inhibited [(3)H]dexamethasone binding to the liver. Modest but statistically significant decreases in brain ex vivo binding were observed with high doses of CORT-108297 [(R)-4α-(ethoxymethyl)-1-(4-fluorophenyl)-6-((4-(trifluoromethyl)phenyl)sulfonyl)-4,4a,5,6,7,8-hexahydro-1H-pyrazolo[3,4-g]isoquinoline] and LLY-2707, but mifepristone inhibited at all doses. Central activity of the GRAs was confirmed by their ability to suppress amphetamine-induced increases in locomotor activity. The increases in the body weight of female rats treated with olanzapine (2 mg/kg PO) over 14 days were reduced in a dose-dependent manner by coadministration of LLY-2707. Similar decreases, although less robust, in body weight were seen with mifepristone and CORT-108297. In addition, sGRAs prevented the glucose excursion after intragastric olanzapine infusions consistent with a direct effect on the hyperglycemia observed during treatment with AAPDs. At doses effectively preventing weight gain, LLY-2707 did not substantially interfere with the dopamine D2 receptor occupancy by olanzapine. Therefore, GRA coadministration may provide a novel treatment modality to prevent the weight gain and diabetes observed during treatment with AAPDs.3.615Whole-blood samples were collected from the arterial catheter using the Culex automated blood sampling system (BASi). Blood samples were collected approximately 30 minutes before pretreatment dosing (T = 0) and at 150 (immediately before the treatment dose), 180, 210, 240, 270, 300, and 330 minutes after the T = 0 sample (eight samples per animal). Samples were processed to plasma and frozen at −80°C until analyzed. Plasma glucose values were determined with a Roche Cobas c311 blood chemistry analyzer (Roche, Basel, Switzerland). Plasma corticosterone was measured by radioimmunoassay (MP Biomedicals, Orangeburg, NY). Plasma insulin was determined using a MSD mouse/rat insulin assay kit (Meso Scale Discovery, Gaithersburg, MD). Data were analyzed using a two-way ANOVA with Bonferroni post hoc testing2014-01-01 00:00:0010.1124/jpet.113.205864
MARS: bringing the automation of small-molecule bioanalytical sample preparations to a new frontierLi, M;Chou, J;Jing, J;Xu, H;Costa, A;Caputo, R;Mikkilineni, R;Flannelly-King, S;Rohde, E;Gan, L;Klunk, L;Yang, L;Pharmacokinetics (PK)Method DevelopmentBioanalysis2012https://www.future-science.com/doi/abs/10.4155/BIO.12.771311-132641122720650In recent years, there has been a growing interest in automating small-molecule bioanalytical sample preparations specifically using the Hamilton MicroLab(®) STAR liquid-handling platform. In the most extensive work reported thus far, multiple small-molecule sample preparation assay types (protein precipitation extraction, SPE and liquid-liquid extraction) have been integrated into a suite that is composed of graphical user interfaces and Hamilton scripts. Using that suite, bioanalytical scientists have been able to automate various sample preparation methods to a great extent. However, there are still areas that could benefit from further automation, specifically, the full integration of analytical standard and QC sample preparation with study sample extraction in one continuous run, real-time 2D barcode scanning on the Hamilton deck and direct Laboratory Information Management System database connectivity. We developed a new small-molecule sample-preparation automation system that improves in all of the aforementioned areas. The improved system presented herein further streamlines the bioanalytical workflow, simplifies batch run design, reduces analyst intervention and eliminates sample-handling error.2.321Rat blood (200 µl each) was collected using a Culex automatic blood-sampling system (BASi, IN, USA) at predose and 5, 15, 30, 45 min, 1, 2, 4, 7, 12, 16 and 24 h after dose into heparinized borosilicate glass tubes, then centrifuged to separate blood cells from plasma2012-06-01 00:00:0010.4155/bio.12.77
Metabolic caging for animal researchKennedy, BW;ADMEData ReviewLab Anim (NY)2012https://www.nature.com/articles/laban0612-171171-176416226140932.794blood, bile, metabolites, dialysates and more from awake and freely moving animals as small as mice and as large as swine," according to the manufacturer's website (http://www.basinc.com/ products/culex). Figure 4: The MetKit rat metabolic cage (BASi) includes a2012-06-01 00:00:0010.1038/laban0612-171
Method for evaluating the potential of C labeled plant polyphenols to cross the blood-brain barrier using accelerator mass spectrometryJanle, EM;Lila, MA;Grannan, M;Wood, L;Higgins, A;Yousef, GG;Rogers, RB;Kim, H;Jackson, GS;Weaver, CM;PK and MicrodialysisNutritional SupplementsNucl Instrum Methods Phys Res B2010https://www.sciencedirect.com/science/article/pii/S0168583X090121051313-13162687-820419067Bioactive compounds in botanicals may be beneficial in preventing age-related neurodegenerative diseases, but for many compounds conventional methods may be inadequate to detect if these compounds cross the blood brain barrier or to track the pharmacokinetics in the brain. By combining a number of unique technologies it has been possible to utilize the power of AMS to study the pharmacokinetics of bioactive compounds in the brain at very low concentrations. (14)C-labeled compounds can be biosynthesized by plant cell suspension cultures co-incubated with radioisotopically-labeled sucrose and isolated and separated into a series of bioactive fractions.To study the pharmacokinetics and tissue distribution of (14)C labeled plant polyphenols, rats were implanted with jugular catheters, subcutaneous ultrafiltration probes and brain microdialysis probes. Labeled fractions were dosed orally. Interstitial fluid (ISF) and brain microdialysate samples were taken in tandem with blood samples. It was often possible to determine (14)C in blood and ISF with a β-counter. However, brain microdialysate samples (14)C levels on the order of 10(7) atoms/sample required AMS technology. The Brain Microdialysate(AUC)/Serum(AUC) ranged from .021- to .029, with the higher values for the glycoside fractions. By using AMS in combination with traditional methods, it is possible to study uptake by blood, distribution to ISF and determine the amount of a dose which can reach the brain and follow the pharmacokinetics in the brain.1.21removed from the probe guide and a 4 mm MD brain probe (Bioanalytical Systems, West Lafayette Sample collection was programmed into the Culex and blood samples were taken at 5, 15 AMS measurements any small differences in the background ratio for each rat brain are 2010-04-01 00:00:0010.1016/j.nimb.2009.10.161
Methods in vascular infusion biotechnology in research with rodentsNolan, TE;Klein, HJ;Pharmacokinetics (PK)Data ReviewILAR J2002https://academic.oup.com/ilarjournal/article-abstract/43/3/175/874686175-18243312105384Infusion of experimental compounds into the vascular system of rodents and the need to collect blood and other biological fluids from small animals comprise an area of emerging importance to biomedical research and drug discovery and development. The advances in the development of transgenic rodents coupled with technical progress in the manufacture and commercial availability of various catheters, swivels, tethers, infusion pumps, and sample collection systems that are described have enabled biomedical scientists to miniaturize vascular infusion and sample collection systems previously used in animal species larger than the rat or mouse. Use of these advanced, miniature vascular infusion systems in rodents is possible only when careful planning of experimental design, expert surgical technique, adequate postoperative care, and fundamental animal welfare considerations are meticulously taken into consideration. Use of these vascular infusion systems in rodents promotes animal welfare and scientific progress through the reduction and refinement of animal models.2.015Most sample collection studies require dual catheters: one for delivery of the test article and the other for collection of blood. This procedure is accomplished in tethered rodents using dual channel swivels. Automated blood collection systems have recently become commercially available. The two systems, Culex from Bioanalytical Systems, Inc., and the Automated Blood Sampler from Instech Solomon, are operated from a computer interface that automatically collects serial samples for up to 24 hr. The Culex system is available for use in rats only. The Instech system can be used for rats and larger laboratory animal species, including dogs and nonhuman primates. Specifications for these systems are available on each company's web site. The major advantages of an automated collection system are accuracy of sample collection, reduced stress from handling of animals, and less technician time involved in collecting sample2002-07-10 00:00:0010.1093/ilar.43.3.175
Microdialysis evaluation of atomoxetine brain penetration and central nervous system pharmacokinetics in ratsKielbasa, W;Kalvass, JC;Stratford, R;PK and MicrodialysisCNSDrug Metab. Dispos.2009http://dmd.aspetjournals.org/content/37/1/137.short137-14237118936112A comprehensive in vivo evaluation of brain penetrability and central nervous system (CNS) pharmacokinetics of atomoxetine in rats was conducted using brain microdialysis. We sought to determine the nature and extent of transport at the blood-brain barrier (BBB) and blood-cerebrospinal fluid barrier (BCB) and to characterize brain extracellular and cellular disposition. The steady-state extracellular fluid (ECF) to plasma unbound (uP) concentration ratio (C(ECF)/C(uP)=0.7) and the cerebrospinal fluid (CSF) to plasma unbound concentration ratio (C(CSF)/C(uP)=1.7) were both near unity, indicating that atomoxetine transport across the BBB and BCB is primarily passive. On the basis of the ratios of whole brain concentration to C(ECF) (C(B)/C(ECF)=170), brain cell (BC) concentration to C(ECF) (C(BC)/C(ECF)=219), and unbound brain cell concentration to C(ECF) (C(uBC)/C(ECF)=2.9), we conclude that whole brain concentration does not represent the concentration in the biophase and atomoxetine primarily partitions into brain cells. The distributional clearance at the BBB (Q(BBB)=0.00110 l/h) was estimated to be 12 times more rapid than that at the BCB (Q(BCB)=0.0000909 l/h) and similar to the clearances across brain parenchyma (CL(ECF-BC)=0.00216 l/h; CL(BC-ECF)=0.000934 l/h). In summary, the first detailed examination using a quantitative microdialysis technique to understand the brain disposition of atomoxetine was conducted. We determined that atomoxetine brain penetration is high, movements across the BBB and BCB occur predominantly by a passive mechanism, and rapid equilibration of ECF and CSF with plasma occurs.3.354The live phase portion of a study was conducted on a Culex Automated Pharmacology System (BAS Bioanalytical Systems) that enabled timed blood and dialysate collections in awake and freely moving animals. Animals were allowed free access to food and water during the time course of sample collection. All activities were performed humanely under a protocol approved by the Eli Lilly and Company Institutional Animal Care and Use Committee in an animal facility accredited by the Association for Assessment and Accreditation of Laboratory Animal Care, International2009-01-01 00:00:0010.1124/dmd.108.023119
Mitigation of sensory and motor deficits by acrolein scavenger phenelzine in a rat model of spinal cord contusive injuryChen, Z;Park, J;Butler, B;Acosta, G;Vega-Alvarez, S;Zheng, L;Tang, J;McCain, R;Zhang, W;Ouyang, Z;Cao, P;Shi, R;Pharmacokinetics (PK)PainJ. Neurochem.2016https://onlinelibrary.wiley.com/doi/abs/10.1111/jnc.13639328-338138227060873Currently there are no effective therapies available for the excruciating neuropathic pain that develops after spinal cord injuries (SCI). As such, a great deal of effort is being put into the investigation of novel therapeutic targets that can alleviate this pain. One such target is acrolein, a highly reactive aldehyde produced as a byproduct of oxidative stress and inflammation that is capable of activating the transient receptor potential ankyrin 1 (TRPA1) cation channel, known to be involved in the transmission and propagation of chronic neuropathic pain. One anti-acrolein agent, hydralazine, has already been shown to reduce neuropathic pain behaviors and offer neuroprotection after SCI. This study investigates another acrolein scavenger, phenelzine, for its possible role of alleviating sensory hypersensitivity through acrolein suppression. The results show that phenelzine is indeed capable of attenuating neuropathic pain behaviors in acute, delayed, and chronic administration schedules after injury in a rat model of SCI. In addition, upon the comparison of hydralazine to phenelzine, both acrolein scavengers displayed a dose-dependent response in the reduction of acrolein in vivo. Finally, phenelzine proved capable of providing locomotor function recovery and neuroprotection of spinal cord tissue when administered immediately after injury for 2 weeks. These results indicate that phenelzine may be an effective treatment for neuropathic pain after SCI and likely a viable alternative to hydralazine. We have shown that phenelzine can attenuate neuropathic pain behavior in acute, delayed, and chronic administration in post-SCI rats. This was accompanied by a dose-dependent reduction in an acrolein metabolite in urine and an acrolein adduct in spinal cord tissue, and the suppression of TRPA1 over-expression in central and peripheral locations post-trauma. Acrolein scavenging might be a novel therapeutic strategy to reduce post-SCI neuropathic pain. © 2016 International Society for Neurochemistry.4.87IN, USA) were implanted in the peritoneal cavity and the carotid artery (BASi Culex rat carotid, Bioanalytical A volume of 100 μL of blood was collected by the Culex sampler and a phosphate‐buffered saline blocking buffer and incubated with a monoclonal mouse anti‐acrolein2016-07-01 00:00:0010.1111/jnc.13639
Modulating the Inflammatory Reflex in Rats Using Low-Intensity Focused Ultrasound Stimulation of the Vagus NerveWasilczuk, KM;Bayer, KC;Somann, JP;Albors, GO;Sturgis, J;Lyle, LT;Robinson, JP;Irazoqui, PP;ADMEInflammatory DiseaseUltrasound Med Biol2019https://www.sciencedirect.com/science/article/pii/S0301562918304022481-48945230396599Tumor necrosis factor α (TNF-α) is linked to several chronic inflammatory diseases. Electrical vagus nerve stimulation reduces serum TNF-α levels but may cause chronic nerve damage and requires surgery. Alternatively, we proposed focused ultrasound stimulation of the vagus nerve (uVNS), which can be applied non-invasively. In this study, we induced an inflammatory response in rats using lipopolysaccharides (LPS) and collected blood to analyze the effects of uVNS on cytokine concentrations. We applied one or three 5-min pulsed focused ultrasound stimulation treatments to the vagus nerve (250 kHz, ISPPA = 3 W/cm2). Animals receiving a single ultrasound application had an average reduction in TNF-α levels of 19%, similar to the 16% reduction observed in electrically stimulated animals. With multiple applications, uVNS therapy statistically reduced serum TNF-α levels by 73% compared with control animals without any observed damage to the nerve. These findings suggest that uVNS is a suitable way to attenuate TNF-α levels. Copyright © 2018 Elsevier Ltd. All rights reserved.2.205Buford, GA, USA) containing K 3 EDTA using the Culex Automatic Blood Collection System (Culex & Honeycomb, Bioanalytical Systems, Inc., West Cytokine levels were determined using a commercial kit (LEGENDplex Rat Th Cytokine Panel 13-plex, BioLegend, San Diego 2019-02-01 00:00:0010.1016/j.ultrasmedbio.2018.09.005
mPEG-PLA/TPGS mixed micelles via intranasal administration improved the bioavailability of lamotrigine in the hippocampusYu, A;Lv, J;Yuan, F;Xia, Z;Fan, K;Chen, G;Ren, J;Lin, C;Wei, S;Yang, F;MicrodialysisEpilepsyInt J Nanomedicine2017https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5701607/8353-83621229200847This study aimed to develop a novel methoxy poly(ethylene glycol)-poly(lactide) (mPEG-PLA)/D-α-tocopherol polyethylene glycol 1000 succinate (TPGS) mixed micelle drug delivery system to improve lamotrigine (LTG) distribution in the hippocampus. LTG-loaded mPEG-PLA/TPGS mixed micelles and LTG-loaded mPEG-PLA micelles were formulated, and their characteristics, particle size, surface morphology, and release behavior in vitro were researched. Then, a microdialysis sampling technique coupled with two validated chromatographic systems was developed for the continuous measurement of the protein-unbound form of LTG in the rat plasma and hippocampus after administering two kinds of micelles and LTG solution intranasally. The drug loading and mean size of LTG-loaded micelles and LTG-loaded mixed micelles prepared with optimal formulation were 36.44%±0.14%, 39.28%±0.26%, 122.9, and 183.5 nm, respectively, with a core-shell structure. The cumulative release rate in vivo of LTG-loaded mixed micelles was 84.21% at 24 hours and showed more sustained release while that of LTG-loaded micelles was 80.61% at 6 hours. The Tmax and area under concentration-time curve from zero to time of last quantifiable concentration of LTG solution, LTG-loaded micelles, and LTG-loaded mixed micelles were 55, 35, and 15 minutes and about 5,384, 16,500, and 25,245 (min⋅μg)/L in the hippocampus, respectively. The results revealed that LTG-loaded mPEG-PLA/TPGS mixed micelles enhanced the absorption of LTG at the nasal cavity and reduced the efflux of LTG in the brain, suggesting that the function of TPGS inhibited P-glycoprotein and LTG-loaded mPEG-PLA/TPGS mixed micelles had the potential to overcome refractory epilepsy.4.471A microdialysis probe was slowly implanted via the guiding cannula into the hippocampus of the rat system consisted of a microdialysis syringe (1 mL, MDN-0100; BASi, Mt Vernon, IN, USA), a Bee syringe pump, and a fraction collector (CULEX fraction collector; BASi)2017-11-21 00:00:0010.2147/IJN.S145488
Natural antioxidants: Their electrochemistry and determinationin plant material and biological fluidsCorredor-Sanchez, PS;PK/PDNutritional SupplementsThesis2007http://search.proquest.com/openview/34655a28f4512e5afbdc6947a70faf41/1?pq-origsite=gscholar&cbl=18750&diss=yValidated LCUVEC and LCMS methods for the determination of curcuminoids, caffeic and chlorogenic acids both in food and in biological fluids permits study of pharmacokinetics. Determination of some primary and secondary metabolites was also achieved. Higher quality pharmacokinetics data was obtained by using an automated pharmacology unit (Culex), thus reducing animal stress during the study2007-01-01 00:00:00
Nilotinib preclinical pharmacokinetics and practical application toward clinical projections of oral absorption and systemic availabilityXia, B;Heimbach, T;He, H;Lin, TH;Pharmacokinetics (PK)OncologyBiopharm Drug Dispos2012https://onlinelibrary.wiley.com/doi/abs/10.1002/bdd.1821536-54933923097199Nilotinib is a highly potent and selective bcr-abl tyrosine kinase inhibitor used for the treatment of patients who are in the chronic and accelerated phases of Philadelphia chromosome-positive (Ph+) chronic myeloid leukemia (CML). Nilotinib preclinical data and its use for practical predictions of systemic exposure profiles and oral absorption are described. The systemic clearance (CL) of nilotinib was relatively low in rodents with a value of less than 25% of hepatic blood flow (Q(H)), while it was moderate in monkeys and dogs (CL/Q(H)  = 32-35%). The steady state volume of distribution (V(ss) ) ranged from 0.55 to 3.9 l/kg across the species tested. The maximum concentration (C(max)) of nilotinib occurred at 0.5-4 h and the bioavailability was moderate (17-44%). The plasma protein binding was high (> 97.5%) in preclinical species and humans. The human CL (~ 0.1 l/h/kg) and V(ss) (~2.0 l/kg) were best predicted by the rat-dog-human proportionality method and allometric scaling method, respectively. The human intravenous pharmacokinetic profile was projected by the Wajima 'C(ss)-MRT' method. The predicted micro-constants from human intravenous profiles were incorporated into the advanced compartmental absorption and transit model within the GastroPlus program to simulate the oral concentration-time curves in humans. Overall, the simulated oral human pharmacokinetic profiles showed good agreement with observed clinical data, and the model predicted that the C(max) , AUC, t(½) , V(z) /F and CL/F values were within 1.3-fold of the observed values. The absolute oral bioavailability of nilotinib in healthy humans was predicted to be low (< 25%). Copyright © 2012 John Wiley & Sons, Ltd.1.611in properly marked metabolism cages placed on an automated sample collection system (Culex Autosampler, BAS Indianapolis, IN) The intravenous plasma concentrations obtained from various species (mouse, rat, dog and monkey) was first normalized by the2012-12-01 00:00:0010.1002/bdd.1821
Oral coadministration of β-glucuronidase to increase exposure of extensively glucuronidated drugs that undergo enterohepatic recirculationEichenbaum, G;Hsu, CP;Subrahmanyam, V;Chen, J;Scicinski, J;Galemmo, RA;Tuman, RW;Johnson, DL;ADMEDrug MetabolismJ Pharm Sci2012https://www.sciencedirect.com/science/article/pii/S00223549153151482545-25561017Extensive first-pass metabolism can significantly limit a drug's oral exposure levels. In this work, we introduce an innovative approach for increasing the oral bioavailability of a drug that undergoes extensive reversible glucuronidation and enterohepatic recirculation through intraduodenal coadministration of the deconjugating enzyme β-glucur-onidase. Intraduodenal administration of JNJ-10198409 (10 mg/kg) with β-glucuronidase (34,000-140,000 units/kg) to catheterized rats resulted in a significant increase (_p_ < 0.005) in the mean area under the plasma concentration versus time curve (AUC; approx. threefold) and maximum plasma concentration (_C_max; approx. twofold) of JNJ-10198409. The AUC and _C_max were 60 ± 18 ng h/mL and 76 ± 29 ng/mL, respectively, with no enzyme and 177 ± 55 ng h/mL and 129 ± 41 ng/mL, respectively, with β-glucuronidase coadministered. Moreover, the AUC of the primary glucuronide metabolite increased approximately sevenfold from 1173 ± 361 (ng h)/mL with no enzyme coadministered to 8723 ± 2133 ng h/mL with coadministered enzyme. These pharmacokinetic data support the hypothesis that when the primary glucuronide is secreted into the duodenum via the bile duct, the glucuronide is converted by β-glucuronidase back to the parent compound. The parent compound is then reabsorbed and reconjugated, resulting in elevated systemic exposures to both parent and glucuronide. Potential clinical and preclinical applications and considerations for this approach are discussed.3.197housed in cages of a Culex TM automated blood sampler (ABS) (Bioanalytical Systems, Inc were collected simultaneously via both cannula and were controlled by the Culex TM ABS tethering system (Instech [Plymouth Meeting, PA]) and housed in the standard rat-size2012-07-01 00:00:0010.1002/jps.23113
P1–Multi-center prevalidation unsing a new in vitro reconstituted human corneal epithelial model to assess the eye irritating potential of chemicals.Alepee, N;Catoire, S;Montbroussou, E;Mignot, A;PK and MicrodialysisMethod Developmentbiotechnocentre.fr2004http://www.biotechnocentre.fr/files/04-actes_colloque.pdf5-5Based on results already published on Blood Brain Barrier passage about morphine and morphine 6-β-D-glucuronide in rat. This experiment was carried out using Culex® and Raturn® to see whether an automatic sampling on awake, non stressed, freely moving animals was easy to set up2004-01-01 00:00:00
Pharmacokinetic and pharmacodynamic properties of SOL1: a novel dual inhibitor of neutral endopeptidase and endothelin converting enzymeNelissen, J;Lemkens, P;Sann, H;Bindl, M;Bassissi, F;Jasserand, D;De Mey, JG;Janssen, BJ;Pharmacokinetics (PK)Blood PressureLife Sci.2012https://www.sciencedirect.com/science/article/pii/S0024320512000707587-5929113-1422365954The pharmacological profile of the novel putative neutral endopeptidase (NEP) and endothelin converting enzyme (ECE) inhibitor SOL1 was examined. The enzyme inhibitory profile of SOL1 was established in vitro. The pharmacokinetic and pharmacodynamic profile was determined in rodents in vivo. In vitro, at neutral pH, 10 μM SOL1 inhibited NEP-1, NEP-2, and ECE-1 by 99%, 94% and 75%, respectively. The IC(50)s were 25, 25 and 3200 nmol/L, respectively. In anesthetized rats, SOL1 inhibited blood pressure (BP) responses to big-ET-1 and ET-1(1-31) with ED(50)s of 1.9 and 0.03 mg/kg, corresponding to plasma EC(50)s of 4.6 and 0.1 μmol/L, respectively. Pharmacokinetics of SOL1 were examined after single injections in mice and rats. In these species, the estimated clearance of SOL1 varied between 5 and 9 ml/kg.min and T(1/2) between 20 and 60 min. Steady state kinetics of SOL1 were examined after continuous s.c. infusions of SOL1 for 3 weeks at 50mg/kg.day in DOCA-salt hypertensive rats. This treatment lowered BP by 22 mmHg. Steady state concentrations of SOL1 in plasma were 3.9 μmol/L. In heart, lung, and kidney the concentrations of SOL1 were 0.4, 1.8, and 20.5 μmol/kg, respectively. About 63% of the daily dose was retrieved unaltered in the urine. These data indicate that SOL1 is primarily a NEP inhibitor in vitro as well as in vivo. Given the preferential renal accumulation and renal clearance of SOL1 additional ECE-1 inhibition in the kidney may have contributed to its chronic BP lowering effects in the DOCA-salt hypertensive rat model. Copyright © 2012 Elsevier Inc. All rights reserved.3.448Venous blood samples were collected regularly using the automated Culex® system (Bioanalytical Systems, Inc, West Lafayette, Indiana, USA) to determine plasma SOL1 concentrations Details on the rat model and surgical protocols can be found in the supplement 2012-10-15 00:00:0010.1016/j.lfs.2012.01.015
Pharmacokinetic and pharmacodynamic studies in rats using a new method of automated blood samplingGunaratna, C;Cregor, M;Kissinger, C;PK/PDMethod DevelopmentCurrent 2000http://www.currentseparations.com/issues/18-4/pdf/chandrani.pdf1-5Headquarters Bioanalytical Systems Inc. 2701 Kent Avenue West Lafayette, IN 47906-1382 *corresponding author: prema@bioanalytical.com F1 The Culex ABS system (foreground) was used to automatically collect whole blood, urine and feces from a single rat over the2000-01-01 00:00:00
Pharmacokinetic evaluation of a 1,3-dicyclohexylurea nanosuspension formulation to support early efficacy assessmentWahlstrom, J;Chiang, P;Ghosh, S;Warren, C;Wene, S;Albin, L;Smith, M;Roberds, S;Pharmacokinetics (PK)Inflammatory DiseaseNanoscale Res Lett2007https://www.biomedcentral.com/openurl?doi=10.1007/s11671-007-9063-7291-29626For the IV bolus dose, the formulation was prepared by diluting the stock nanosuspension preparation (10 mg/mL) with 1% (w/w) Cremophore EL in phosphate buffered saline until the desired concentration was reached. For the IV infusion dose, the formulation was prepared by diluting the stock nanosuspension solution (10 mg/mL) to twice the desired target concentration by diluting with 1% (w/w) Cremophore EL in phosphate buffered saline and then further diluting with 1% (w/w) Cremophore EL in phosphate buffered saline containing 20% PVP (w/w). Final solutions were tested for concentration using the analytical methods described below. The total dissolution time and risk assessment for the IV formulation was conducted by measuring the plasma solubility of DCU, particle size of the nanoparticles and the diffusion coefficient of DCU in rat plasma. Both Noyes–Whitney and Hixson–Crowell cube root laws were used in the calculation in a conservative way. The calculation was conducted by assuming the diffusion layer thickness is close to the mean particle size and all particles are uniformly sized. The complete dissolution time for each particle was estimated to be less than a minute upon injection [22 [/articles/10.1007/s11671-007-9063-7#ref-CR22]]. The dissolution time was calculated without the considering the advantage of the turbulent blood flow in the vein, which should serve to further reduce the diffusion boundary thickness, rapidly disperse the initial injection volume and minimize local concentration effects. Thus, the DCU nanoparticle formulation is expected to have instant and complete dissolution in the blood upon the IV injection and infusion.3.159total dissolution time and risk assessment for the IV formulation was conducted by measuring the plasma solubility of DCU, particle size of the nanoparticles and the diffusion coefficient of DCU in rat plasma Animals were acclimated in Culex cages (BASi) overnight prior2007-06-25 00:00:0010.1007/s11671-007-9063-7
Pharmacokinetics and tissue distribution of 14C-labeled grape polyphenols in the periphery and the central nervous system following oral administrationJanle, EM;Lila, MA;Grannan, M;Wood, L;Higgins, A;Yousef, GG;Rogers, RB;Kim, H;Jackson, GS;Ho, L;Weaver, CM;ADMENutritional SupplementsJ Med Food2010https://www.liebertpub.com/doi/abs/10.1089/jmf.2009.0157926-93313420673061Grape polyphenols confer potential health benefits, including prevention of neurodegenerative diseases. To determine the absorption and tissue distribution of the complex grape polyphenol mixture, (14)C-labeled polyphenols were biosynthesized by grape cell suspension cultures, during co-incubation with radioisotopically labeled sucrose, and fractionated into polyphenolic subfractions. The pharmacokinetics and distribution of grape polyphenols into blood, brain, and peripheral interstitial fluid were determined by tracking the (14)C label. The blood peak (14)C concentration of the fractions ranged from 15 minutes to 4 hours. Absorption and tissue distribution varied greatly between fractions. Concentrations in interstitial fluid were lower than in blood. The amount of residual label in the brain at 24 hours ranged from 0.1% to 1.7% of the dose, depending on the fraction. (14)C label found in the brain tissue and brain microdialysate indicated that grape polyphenols or their metabolites are able to cross the blood-brain barrier. Using (14)C-labeled plant polyphenols it is possible to track the compounds or their metabolic products into any tissue and determine distribution patterns in spite of low concentrations. A central question regarding the potential role of dietary polyphenolics in neurodegenerative research is whether they are bioavailable in the brain. Our observations indicate that some grape-derived polyphenolics do reach the brain, which suggests their potential value for applications in neurodegenerative disorders.2.02al effects have been demonstrated even though the polyphenol was not found in the target tissue. Feeding resveratrol reduced the amyloid plaque formation associated with neurodegenerative diseases in mouse brains even though no resveratrol or metabolites could be detected in the brain using HPLC detection methods.23 14C-labeled compounds have long been used to investigate bioavailability of drugs ... achieve optimal efficacy. This synergistic effect of multiple components of a botanical was demonstrated by Morré _et al_.21 in their investigations of the effect of green tea catechins on cancer. Epigallocatechin gallate (10−5 _M_) is effective in inhibiting the growth of HeLa cells in culture. Epicatechin (10−4 _M_) alone has no inhibitory power but when added to epigallocatechin gallate i ... ubmed/19041676]] [Google Scholar] 24. Cotler S. Bugge CJ. Colburn WA. Role of gut contents, intestinal wall, and liver on the first pass metabolism and absolute bioavailability of isotretinoin in the dog. Drug Metab Dispos. 1983;11:458–462. [PubMed [/pubmed/6138231]] [Google Scholar [https://scholar.google.com/scholar_lookup?journal=Drug+Metab+Dispos&title=Role+of+gut+contents,+intestinal+wall,+;... 2010-08-01 00:00:0010.1089/jmf.2009.0157
Pharmacokinetics of DS-96, an alkylpolyamine lipopolysaccharide sequestrant, in rodentsShrestha, A;Li, R;Sil, D;Pardeshi, NN;Schwarting, N;Schorno, KS;Rajewski, RA;Datta, A;David, SA;Pharmacokinetics (PK)Septic ShockJ Pharm Sci2008http://dx.doi.org/10.1002/jps.213615376-5385971218383338The pharmacokinetics of DS-96, an N-alkylhomospermine analog designed to sequester bacterial lipopolysaccharides, has been determined in rodent species. The elimination half-life in mice and rats are about 400 and 500 min, respectively, with other PK parameters being quite similar in the two rodent species. Interestingly, the mouse intravenous plasma concentration time curves exhibit an apparent absorption phase. While the rat intravenous data did not exhibit a pronounced apparent absorption phase immediately following injection, plasma levels did increase between 10 and 30 min following an expected drop from time 0 to 5 min. The data are consistent with first-pass uptake, possibly by the lung, with back diffusion as a function of time. The observed C(max) values of 1.36 microg/mL in the mouse intraperitoneal model suggest that a plasma concentration of 0.5-1 microg/mL corresponds to complete protection for a 200 ng/animal dose of intraperitoneally administered LPS in the D-galactosamine-primed model of endotoxin-induced lethality.3.197Adult male Sprague–Dawley rats (230–254 g; Charles River), precannulated with indwelling catheters in the carotid artery and jugular vein for sampling and dosing, respectively, were housed during the experiment in rat metabolic cages of a Culex Automated Pharmacology System (Bioanalytical Systems, Inc., West Lafayette, IN).31,32 The animals (n ¼ 4) were injected i.v. via the jugular vein cannula with a dose of 12 mg/kg of DS-96. Eighty microliters blood samples were collected at preprogrammed intervals from the carotid artery by the Culex system, and stored temporarily on the onboard MD-1202 fraction collector at 48C. Plasma was obtained by centrifugation at 3000 RPM and stored at 808C2008-12-01 00:00:0010.1002/jps.21361
Pharmacokinetics of green tea catechins in extract and sustained-release preparationsJanle, EM;Morré, DM;Morré, DJ;Zhou, Q;Zhu, Y;PK and MicrodialysisNutritional SupplementsJ Diet Suppl2008https://www.tandfonline.com/doi/abs/10.1080/19390210802414279248-2635319885387Catechins are a major constituent of green tea. For green tea to have cancer therapeutic benefit, catechin concentrations in the range of 100 nM are required continuously until apoptosis (programmed cell death) is induced. To prolong elevated plasma and interstitial concentrations of catechins, a sustained-release formulation of green tea extract was tested and compared to a commercial green tea extract (Tegreen97®). Sustained-release formulations are usually developed in the pharmaceutical industry to slowly deliver the compound over a period of time and increase the dosing interval. Plasma and interstitial fluid (ISF) pharmacokinetics of catechins were determined following an oral dose in the rat. The sustained-release formulation profile included multiple smaller peaks of total catechins in both plasma and ISF. Interstitial fluid profiles of green tea extract indicate that higher catechins concentration and longer duration in tissue than in blood may make a sustained-release form unnecessary.Ten male Sprague Dawley rats between 250 and 300 g were obtained from Harlan (Indianapolis, IN). The rats were anesthetized with a mixture of ketamine and xylazine (10:1 100 mg/ml) at a dose of 0.1 ml/100 g and implanted with jugular catheters and subcutaneous ultrafiltration probes (PN MF-7023, Bioanalytical Systems, INC, West Lafayette, IN) under aseptic conditions as previously described (Janle, Portocarrero, Zhu, & Zhou, 2005). The rats were placed in a Culex-automated blood sampling system (Bioanalytical Systems, INC. West Lafayette, IN) and allowed to recover from surgery for 2 days. During the recovery period the Culex was placed on the “Tend” protocol which flushes the catheter with small amounts of sterile saline at 10 minute intervals to maintain patency. The ultrafiltration probe was connected to a needle hub which was inserted into a Vacutainer® tube to maintain flow until the dosing study. Before dosing, the ultrafiltration probe was attached to a mini peristaltic pump and fractions were collected in a fraction collector (BAS, West Lafayette, IN)2008-11-04 00:00:0010.1080/19390210802414279
Pharmacokinetics, Distribution, Metabolism, and Excretion of Omadacycline following a Single Intravenous or Oral Dose of 14C-Omadacycline in RatsLin, W;Flarakos, J;Du, Y;Hu, W;He, H;Mangold, J;Tanaka, SK;Villano, S;Pharmacokinetics (PK)AntibioticsAntimicrob. Agents Chemother.2017https://aac.asm.org/content/61/1/e01784-16.short61127821446The absorption, distribution, metabolism, and excretion (ADME) of omadacycline, a first-in-class aminomethylcycline antibiotic with a broad spectrum of activity against Gram-positive, Gram-negative, anaerobic, and atypical bacteria, were evaluated in rats. Tissue distribution was investigated by quantitative whole-body autoradiography in male Long-Evans Hooded (LEH) rats. Following an intravenous (i.v.) dose of 5 mg/kg of body weight, radioactivity widely and rapidly distributed into most tissues. The highest tissue-to-blood concentration ratios (t/b) were observed in bone mineral, thyroid gland, and Harderian gland at 24 h post-i.v. dose. There was no evidence of stable accumulation in uveal tract tissue, suggesting the absence of a stable binding interaction with melanin. Following a 90 mg/kg oral dose in LEH rats, the highest t/b were observed in bone mineral, Harderian gland, liver, spleen, and salivary gland. The plasma protein binding levels were 26% in the rat and 15% to 21% in other species. Omadacycline plasma clearance was 1.2 liters/h/kg, and its half-life was 4.6 h; the steady-state volume of distribution (Vss) was 6.89 liters/kg. Major circulating components in plasma were intact omadacycline and its epimer. Consistent with observations in human, approximately 80% of the dose was excreted into the feces as unchanged omadacycline after i.v. administration. Fecal excretion was primarily the result of biliary excretion (∼40%) and direct gastrointestinal secretion (∼30%). However, urinary excretion (∼30%) was equally prominent after i.v. dosing. Copyright © 2016 Lin et al.4.715p (n = 3) received 14C-omadacycline at 5 mg/kg (20.0 μCi/mg) by slow bolus injection via a jugular catheter. Approximately 250 μl of blood was collected using an automatic sample collection system (Culex; BASi, West Lafayette, IN). The samples were delivered to preheparinized tubes in 4°C sample collection carousels and stored until further processing. Saline solution (250 μl) was automaticall;... 3) received 14C-omadacycline at 5 mg/kg (20.0 μCi/mg) by slow bolus injection via a jugular catheter. Approximately 250 μl of blood was collected using an automatic sample collection system (Culex; BASi, West Lafayette, IN). The samples were delivered to preheparinized tubes in 4°C sample collection carousels and stored until further processing. Saline solution (250 μl) was automatically injec ... the omadacycline concentration range of 10 to 10,000 ng/ml, no obvious concentration dependency of plasma protein binding was found. The mean unbound protein fractions in plasma were 84.7% ± 5.3% in mouse, 73.9% ± 12.1% in rat, 78.8% ± 7.3% in monkey, and 78.7% ± 9.7% in human. PHARMACOKINETICS, METABOLISM, AND EXCRETION OF OMADACYCLINE. Following a 90 mg/kg oral dose of 14C-omadacycline, t ... TISSUE AND BODY FLUID DISTRIBUTION VIA QWBA. Distribution of radioactivity of omadacycline into tissues, organs, and body fluids was investigated by quantitative whole-body autoradiography (QWBA). Pigmented Long-Evans Hooded (LEH) rats were administered a single oral dose (_n_ = 8) of 14C-omadacycline at 90 mg/kg (1.92 μCi/mg) via gavage and a single i.v. dose (_n_ = 2) of 14C-omadacycline at ... 0,000 ng/ml, no obvious concentration dependency of plasma protein binding was found. The mean unbound protein fractions in plasma were 84.7% ± 5.3% in mouse, 73.9% ± 12.1% in rat, 78.8% ± 7.3% in monkey, and 78.7% ± 9.7% in human. PHARMACOKINETICS, METABOLISM, AND EXCRETION OF OMADACYCLINE. Following a 90 mg/kg oral dose of 14C-omadacycline, the peak radioactivity concentration in plasma ( ... Daley-Yates PT, Feenstra KL, Koike S, Ozawa N, Peng GW, Sams JP, Schuette MR, Yamazaki S. 2002. Pharmacokinetics, toxicokinetics, distribution, metabolism and excretion of linezolid in mouse, rat and dog. Xenobiotica 32:907–924. doi:10.1080/00498250210158249. [PubMed [/pubmed/12419019]] [CrossRef [//dx.doi.org/10.1080%2F00498250210158249]] [Google Scholar [https://scholar.google.com/scholar_look;... 3) received 14C-omadacycline at 5 mg/kg (20.0 μCi/mg) by slow bolus injection via a jugular catheter. Approximately 250 μl of blood was collected using an automatic sample collection system (Culex; BASi, West Lafayette, IN). The samples were delivered to preheparinized tubes in 4°C sample collection carousels and stored until further processing. Saline solution (250 μl) was automatically injec ... the omadacycline concentration range of 10 to 10,000 ng/ml, no obvious concentration dependency of plasma protein binding was found. The mean unbound protein fractions in plasma were 84.7% ± 5.3% in mouse, 73.9% ± 12.1% in rat, 78.8% ± 7.3% in monkey, and 78.7% ± 9.7% in human. Pharmacokinetics, metabolism, and excretion of omadacycline. Following a 90 mg/kg oral dose of 14C-omadacycline, the ... . Tissue and body fluid distribution via QWBA. Distribution of radioactivity of omadacycline into tissues, organs, and body fluids was investigated by quantitative whole-body autoradiography (QWBA). Pigmented Long-Evans Hooded (LEH) rats were administered a single oral dose (n = 8) of 14C-omadacycline at 90 mg/kg (1.92 μCi/mg) via gavage and a single i.v. dose (n = 2) of 14C-omadacycline at 5 mg ... 0,000 ng/ml, no obvious concentration dependency of plasma protein binding was found. The mean unbound protein fractions in plasma were 84.7% ± 5.3% in mouse, 73.9% ± 12.1% in rat, 78.8% ± 7.3% in monkey, and 78.7% ± 9.7% in human. Pharmacokinetics, metabolism, and excretion of omadacycline. Following a 90 mg/kg oral dose of 14C-omadacycline, the peak radioactivity concentration in plasma (Cm ... , ChibaK, Daley-YatesPT, FeenstraKL, KoikeS, OzawaN, PengGW, SamsJP, SchuetteMR, YamazakiS 2002 Pharmacokinetics, toxicokinetics, distribution, metabolism and excretion of linezolid in mouse, rat and dog. Xenobiotica 32:907–924. doi:10.1080/00498250210158249.12419019 12. SlatterJG, SamsJP, EasterJA, FateGD, ChibaK, JohnsonMG, CourtneyM, KoetsMD, NorrisLR, JonesBW 2003 Assessment of radioactive r2017-01-01 00:00:0010.1128/AAC.01784-16
Pharmacokinetics, safety, and hydrolysis of oral pyrroloquinazolinediamines administered in single and multiple doses in ratsLi, Q;Kozar, MP;Shearer, TW;Xie, LH;Lin, AJ;Smith, KS;Si, Y;Anova, L;Zhang, J;Milhous, WK;Skillman, DR;Pharmacokinetics (PK)AntimalarialAntimicrob. Agents Chemother.2007https://aac.asm.org/content/51/8/2898.short2898-290451817562804Pyrroloquinazolinediamine (PQD) derivatives such as tetra-acetamide PQD (PQD-A4) and bis-ethylcarbamyl PQD (PQD-BE) were much safer (with therapeutic indices of 80 and 32, respectively) than their parent compound, PQD (therapeutic index, 10). Further evaluation of PQD-A4 and PQD-BE in single and multiple pharmacokinetic (PK) studies as well as corresponding toxicity studies was conducted with rats. PQD-A4 could be converted to two intermediate metabolites (monoacetamide PQD and bisacetamide PQD) first and then to the final metabolite, PQD, while PQD-BE was directly hydrolyzed to PQD without precursor and intermediate metabolites. Maximum tolerant doses showed that PQD-A4 and PQD-BE have only 1/12 and 1/6, respectively, of the toxicity of PQD after a single oral dose. Compared to the area under the concentration-time curve for PQD alone (2,965 ng.h/ml), values measured in animals treated with PQD-A4 and PQD-BE were one-third (1,047 ng.h/ml) and one-half (1,381 ng.h/ml) as high, respectively, after an equimolar dosage, suggesting that PQD was the only agent to induce the toxicity. Similar results were also shown in multiple treatments; PQD-A4 and PQD-BE generated two-fifths and three-fifths, respectively, of PQD concentrations, with 8.8-fold and 3.8-fold safety margins, respectively, over the parent drug. PK data indicated that the bioavailability of oral PQD-A4 was greatly limited at high dose levels, that PQD-A4 was slowly converted to PQD via a sequential three-step process of conversion, and that PQD-A4 was significantly less toxic than the one-step hydrolysis drug, PQD-BE. It was concluded that the slow and smaller release of PQD was the main reason for the reduction in toxicity and that the active intermediate metabolites can still maintain antimalarial potency. Therefore, the candidate with multiple-step hydrolysis of PQD could be developed as a safer potential agent for malaria treatment.4.7151-3, PQD (ng·h/ml) 1,903.5 ± 241.1 2,856.1 ± 339.7 4,724.6 ± 684.2     AUCD3/AUCD1 1.86 ± 0.69 1.77 ± 0.51 1.66 ± 0.57 a Blood samples were collected by using a Culex automated blood sampler. b PQD bioavailabilities were calculated via total AUCoral (PQD)/AUCi.v. (PQD) (i.v., intravenous), with adjustment to the same dose level. c MW, molecular weight. d N;Serial plasma samples were collected during the absorption, distribution, and elimination phases for full PK analysis. Each animal was dosed, and plasma samples were obtained for up to 2 days. A total of 11 (at 0, 30, and 60 min and 2, 3, 5, 8, 12, 24, 36, and 48 h) plasma samples were collected into cooled vials by using a Culex automated blood sampler (Bioanalytical Systems Inc., West Lafayette, IN). One hundred microliters of blood per sample for intravenous groups and 200 μl for oral gavage groups were collected for all samples. During collection, blood samples were mixed with heparin-saline (50 μl) to obtain a total volume of 150 μl for intravenous samples and 250 μl for intragastric samples. All samples were analyzed by using LC-MS/MS. The Culex automatic blood sampler is a blood sampling and metabolic monitoring machine, which is controlled by its own internal computer. By using this system we were successful in evaluating PK for long-term periods. The detailed sampling process was described previously (18)2007-08-01 00:00:0010.1128/AAC.00932-06
Preclinical pharmacodynamic and pharmacokinetic characterization of the major metabolites of cariprazineKiss, B;Némethy, Z;Fazekas, K;Kurkó, D;Gyertyán, I;Sághy, K;Laszlovszky, I;Farkas, B;Kirschner, N;Bolf-Terjéki, E;Balázs, O;Lendvai, B;MicrodialysisAntipsychoticDrug Des Devel Ther2019https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6754336/3229-32481331571826Cariprazine, a dopamine D3-preferring D3/D2 receptor partial agonist and serotonin 5-HT1A receptor partial agonist, has two major human metabolites, desmethyl-cariprazine (DCAR) and didesmethyl-cariprazine (DDCAR). The metabolite pharmacology was profiled to understand the contribution to cariprazine efficacy. In vitro receptor binding and functional assays, electrophysiology, animal models, microdialysis, and kinetic-metabolism approaches were used to characterize the pharmacology of DCAR and DDCAR. Similar to cariprazine, both metabolites showed high affinity for human D3, D2L, 5-HT1A, 5-HT2A, and 5-HT2B receptors, albeit with higher selectivity than cariprazine for D3 versus D2 receptors. In [35S]GTPγS binding assays, cariprazine and DDCAR were antagonists in membranes from rat striatum and from cells expressing human D2 and D3 receptors, and were partial agonists in membranes from rat hippocampus. In cAMP signaling assays, cariprazine, DCAR, and DDCAR acted as partial agonists at D2 and D3 receptors; cariprazine and DDCAR were full agonists, whereas DCAR was a partial agonist at 5-HT1A receptors. Cariprazine, DCAR, and DDCAR were pure antagonists at human 5-HT2B receptors. Cariprazine and DDCAR increased rat striatal dopamine and reduced cortical serotonin turnover. Cariprazine and DDCAR showed similar in vivo D3 receptor occupancy in rat brain; however, cariprazine was more potent for D2 receptor occupancy. Both cariprazine and DDCAR dose-dependently but partially suppressed the spontaneous activity of midbrain dopaminergic neurons in rats, with the parent compound being more potent but shorter acting than its metabolite. Consistent with the D2 receptor occupancy profile, DDCAR was 3- to 10-fold less potent than cariprazine in rodent models of antipsychotic-like activity. Following acute cariprazine administration, DDCAR was detected in the rodent brain but at much lower levels than cariprazine. Overall, in vitro and in vivo pharmacological profiles of DCAR and DDCAR demonstrated high similarity with cariprazine, suggesting that the major metabolites of cariprazine contribute significantly to its clinical efficacy. © 2019 Kiss et al.3.208A dual-probe MD study was conducted to estimate the free extracellular levels of cariprazine and DDCAR in awake, freely moving, male Wistar rats (n=5). Cariprazine was administered intragastrically at 1 mg/kg dose in d.w. via surgically implanted catheter. The MD experiments were performed with the Culex automated sampling system (Bioanalytical Systems Inc.). Probes were implanted in the prefrontal cortex and striatum and perfused with artificial cerebrospinal fluid at a flow rate of 1 µL/min. Dialysate samples were collected in 20-min intervals from 60 min pre-dose to ≥300 min post-dose. Dialysate concentrations of cariprazine and DDCAR were determined by LC-MS/MS2019-09-16 00:00:0010.2147/DDDT.S188760
Prevention of fostamatinib-induced blood pressure elevation by antihypertensive agentsLengel, D;Lamm Bergström, E;Barthlow, H;Oldman, K;Musgrove, H;Harmer, A;Valentin, JP;Duffy, P;Braddock, M;Curwen, J;Safety PharmacologyRheumatoid ArthritisPharmacol Res Perspect2015https://bpspubs.onlinelibrary.wiley.com/doi/abs/10.1002/prp2.176e001763526516588Fostamatinib is a tyrosine kinase inhibitor with activity against spleen tyrosine kinase which has completed clinical trials for patients with rheumatoid arthritis. In clinical studies fostamatinib treatment was associated with a small elevation of systemic arterial blood pressure (BP), a similar finding to that seen with other kinase inhibitors, especially those that inhibit VEGFR2 signaling. We have investigated the link between fostamatinib-induced blood pressure elevation and plasma levels of the fostamatinib-active metabolite R940406 in conscious rats and found the time course of the BP effect correlated closely with changes in R940406 plasma concentration, indicating a direct pharmacological relationship. Free plasma levels of R940406 produced in these studies (up to 346 nmol/L) span the clinically observed mean peak free plasma concentration of 49 nmol/L. We have demonstrated that the blood pressure elevation induced by fostamatinib dosing can be successfully controlled by a variety of methods, notably simple drug withdrawal or codosing with a range of standard antihypertensive agents such as atenolol, captopril, and nifedipine. These findings support potential methods of maintaining patient safety while on fostamatinib therapy. Furthermore, we have demonstrated, using nifedipine as an example agent, that this blood pressure control was not achieved by reduction in plasma exposure of R940406, suggesting that potential benefits from the pharmacology of the investigational drug can be maintained while blood pressure control is managed by use of standard comedications.mm into the inferior vena cava. The cannula was used during the experimental phase of the study to obtain blood samples using an automated blood sampler (Culex, BASi, West Lafayette, IN). Rats were then shipped to AstraZeneca ; Rats were exposed to a 12‐h light:dark cycle and given full ad libitum access to a standard rat chow (Purina Labdiet 5001 The cannula was used during the experimental phase of the study to obtain blood samples using an automated blood sampler (Culex, BASi, West Lafayette2015-10-01 00:00:0010.1002/prp2.176
Probing a potential in vivo drug-excipient interaction: temporal effects of a modified β-cyclodextrin on the intravenous pharmacokinetics of a model high-affinity drug ligandMannila, A;Morizzi, J;Nguyen, TT;Charman, SA;McIntosh, MP;Shackleford, DM;PK/PDDrug/Drug InteractionsJ Pharm Sci2012https://www.sciencedirect.com/science/article/pii/S00223549153144773381-3389101922549698The investigational synthetic ozonide, OZ209, has previously been shown to have high binding affinity for sulfobutylether(7)-β-cyclodextrin [(SBE)(7)-β-CD] resulting in altered pharmacokinetics when administered intravenously to rats in a (SBE)(7)-β-CD aqueous formulation. In the present study, OZ209 and (SBE)(7)-β-CD have been used to probe whether a modified β-CD excipient, on systemic administration, can bind to and alter the pharmacokinetics of a coadministered drug. When (SBE)(7)-β-CD was administered 60 min after OZ209, a spike in the concentration of OZ209 in blood and plasma was detected within 2 min of the (SBE)(7)-β-CD infusion, and this was accompanied by a temporary decrease in the whole blood-to-plasma partitioning ratio of OZ209, the duration of which was dependent upon the dose of (SBE)(7)-β-CD. Administration of (SBE)(7)-β-CD also resulted in increased urinary excretion of OZ209. By contrast, administration of (SBE)(7)-β-CD 4 h prior to OZ209 had no pronounced effect on the blood or plasma pharmacokinetics of OZ209, consistent with the (SBE)(7)-β-CD having been largely eliminated prior to the administration of OZ209. This study is the first to demonstrate an in vivo drug-excipient interaction between a modified β-CD and a coadministered drug, and also demonstrates that such an interaction can be avoided through appropriate consideration of CD pharmacokinetics. Copyright © 2012 Wiley Periodicals, Inc.3.19711 Briefly, blood samples (170 μL) from awake and freely moving rats were collected using the Culex ABS (BASi) A 100 μL aliquot of urine was taken every time the rat voided, noting the time and volume of the urine collected2012-09-01 00:00:0010.1002/jps.23177
Protective effect of nectandrin B, a potent AMPK activator on neointima formation: inhibition of Pin1 expression through AMPK activationKi, SH;Lee, JW;Lim, SC;Hien, TT;Im, JH;Oh, WK;Lee, MY;Ji, YH;Kim, YG;Kang, KW;Pharmacokinetics (PK)Vascular DiseaseBr. J. Pharmacol.2013https://bpspubs.onlinelibrary.wiley.com/doi/abs/10.1111/j.1476-5381.2012.02228.x932-945168423004677Neointima is considered a critical event in the development of vascular occlusive disease. Nectandrin B from nutmeg functions as a potent AMP-activated protein kinase (AMPK) activators. The present study addressed whether nectandrin B inhibits intimal hyperplasia in guide wire-injured arteries and examined its molecular mechanism. Neointima was induced by guide wire injury in mouse femoral arteries. Cell proliferation and mechanism studies were performed in rat vascular smooth muscle cells (VSMC) culture model. Nectandrin B increased AMPK activity in VSMC. Nectandrin B inhibited the cell proliferation induced by PDGF and DNA synthesis. Moreover, treatment of nectandrin B suppressed neointima formation in femoral artery after guide wire injury. We have recently shown that Pin1 plays a critical role in VSMC proliferation and neointima formation. Nectandrin B potently blocked PDGF-induced Pin1 and cyclin D1 expression and nectandrin B's anti-proliferation effect was diminished in Pin1 overexpressed VSMC. PDGF-induced phosphorylation of ERK and Akt was marginally affected by nectandrin B. However, nectandrin B increased the levels of p53 and its downstream target p21 and, also reversibly decreased the expression of E2F1 and phosphorylated Rb in PDGF-treated VSMC. AMPK inhibition by dominant mutant form of adenovirus rescued nectandrin B-mediated down-regulation of Pin1 and E2F1. Nectandrin B inhibited VSMC proliferation and neointima formation via inhibition of E2F1-dependent Pin1 gene transcription, which is mediated through the activation of an AMPK/p53-triggered pathway. © 2012 The Authors. British Journal of Pharmacology © 2012 The British Pharmacological Society.6.583Apparatus and LC‐MS/MS conditions for pharmacokinetics of nectandrin B. The automated blood sampling unit consisted of a freely moving mouse containment device (RaturnTM, BASi, West Lafayette, IN, USA) and an automated blood sampler Culex™ (BASi) ; VSMC were isolated from rat thoracic aorta as described previously (Lee et al., 2009) The automated blood sampling unit consisted of a freely moving mouse containment device (RaturnTM, BASi, West Lafayette, IN, USA) and an automated blood sampler Culex™ (BASi)2013-02-01 00:00:0010.1111/j.1476-5381.2012.02228.x
Quantitative analysis of PD 0332991 in mouse plasma using automated micro-sample processing and microbore liquid chromatography coupled with tandem mass spectrometrySmith, D;Tella, M;Rahavendran, SV;Shen, Z;Pharmacokinetics (PK)OncologyJ. Chromatogr. B Analyt. Technol. Biomed. Life Sci.2011https://pdfs.semanticscholar.org/7cf2/179eb7a42ac2d59dd5930dd96cb0c197b88e.pdf2860-28658792721889427In the oncology therapeutic area, the mouse is the primary animal model used for efficacy studies. Often with mouse pharmacokinetic (PK) and pharmacokinetic/pharmacodynamic (PK/PD) studies, less than 20 μL of total plasma sample volume is available for bioanalysis due to the small size of the animal and the need to split samples for other measurements such as biomarker analyses. The need to conduct automated "small volume" sample processing for quantitative bioanalysis has therefore increased. An automated fit for purpose protein precipitation (PPT) method using a Hamilton MicroLab Star (Reno, NV, USA) to support mouse PK and PK/PD studies for an oncology drug candidate PD 0332991, (a specific inhibitor of cyclin-dependent kinase 4 (CDK-4) currently in development) for processing "small volumes" was developed. The automated PPT method was achieved by extracting and processing 10 μL out of a minimum sample volume of 15 μL plasma utilizing the Hamilton MicroLab Star. A 96-conical shallow well plate by Agilent Technologies, Inc (Wilmington, DE, USA) was the labware of choice used in the automated Hamilton "small volume" method platform. Analyses of a 10 μL plasma aliquot from 15 μL of plasma study samples were conducted by both automated and manual PPT method. All plasma samples were quantitated using a Sciex API 4000 triple quadrupole mass spectrometer coupled with an Eksigent Express HT Ultra HPLC system. The chromatography was achieved using an Agilent microbore C(18) Extend, 1.0 × 50 mm, 3.5 μm column at a flow rate of 0.150 mL/min with a total run time of 1.8 min. Accuracy and precision of standard and QC concentration levels were within 90-107% and <14%, respectively. Calibration curves were linear over the dynamic range of 1.0-1000 ng/mL. PK studies for PD 0332991 were conducted in female C3H mice following intravenous administration at 1mg/kg and oral administration at 2mg/kg. PK values such as area under curve (AUC), volume of distribution (Vd), clearance (Cl), half life (T(1/2)) and bioavailability (F%) demonstrated less than 11% difference between the automated Hamilton and manual PPT methods. The results demonstrate that the automated Hamilton PPT method can accurately and precisely aliquot 10 μL of plasma from 15 μL or larger volume plasma samples. The fit for purpose Hamilton PPT method is suitable for routine analyses of plasma samples from micro-sampling PK and PK/PD samples to support discovery studies. Copyright © 2011 Elsevier B.V. All rights reserved.2.813The main players on the market are Verutech (Accusampler), Basi (Culex) and Instech (ABS2) 12. Bundgaard C, Jorgensen M, Mork A. An integrated microdialysis rat model for multiple pharmacokinetic/pharmacodynamic investigations of serotonergic agents2011-10-01 00:00:0010.1016/j.jchromb.2011.08.009
Quantitative bioanalytical and analytical method development of dibenzazepine derivative, carbamazepine: A reviewDatar, PA;PharmacologyData ReviewJ Pharm Anal2015https://www.sciencedirect.com/science/article/pii/S2095177915000337213-2225429403934Bioanalytical methods are widely used for quantitative estimation of drugs and their metabolites in physiological matrices. These methods could be applied to studies in areas of human clinical pharmacology and toxicology. The major bioanalytical services are method development, method validation and sample analysis (method application). Various methods such as GC, LC-MS/MS, HPLC, HPTLC, micellar electrokinetic chromatography, and UFLC have been used in laboratories for the qualitative and quantitative analysis of carbamazepine in biological samples throughout all phases of clinical research and quality control. The article incorporates various reported methods developed to help analysts in choosing crucial parameters for new method development of carbamazepine and its derivatives and also enumerates metabolites, and impurities reported so far.4.44Samples were injected by an autosampler (Sample Sentinel, BASi), which was set at 10 °C and The extraction recovery of analytes from rat plasma was over 87 techniques (ie manual sampling via a jugular catheter versus automated sampling by using Culex automated blood ; Samples were injected by an autosampler (Sample Sentinel, BASi), which was set at 10 °C and The extraction recovery of analytes from rat plasma was over 87 techniques (ie manual sampling via a jugular catheter versus automated sampling by using Culex automated blood ; Samples were injected by an autosampler (Sample Sentinel, BASi), which was set at 10 °C and The extraction recovery of analytes from rat plasma was over 87 techniques (ie manual sampling via a jugular catheter versus automated sampling by using Culex automated blood ;... v/v/v). The flow rate was maintained at 1.0 mL/min. Mass spectral data showed protonated molecular ion peaks at m/z 237, m/z 196 and m/z 501 for CBZ, impurity-A and impurity-B, respectively. On the basis of RRT and mass spectral data, impurity-A having molecular ion peak at m/z 196 was identified as iminodibenzyl. The mass spectral data obtained for impurity-B at RT 95.8 min did not match with ... y a quick centrifugation. Without any change to a conventional HPLC system, CBZ and CBZ-EP were well separated in less than 2.5 min using a Kovasil MS C14 column. No interference was observed with endogenous compounds and with nine antiepileptic drugs commonly prescribed as co-medication, and their metabolites. Due to the very low specific surface area of the packing, the required organic modifie; Samples were injected by an autosampler (Sample Sentinel, BASi), which was set at 10 °C and The extraction recovery of analytes from rat plasma was over 87 techniques (ie manual sampling via a jugular catheter versus automated sampling by using Culex automated blood2015-08-01 00:00:0010.1016/j.jpha.2015.02.005
Rat BiodataHeiser, A;Liu, J;Drug DevelopmentMethod DevelopmentRat Jugular Vein and Carotid Artery Catheterization for Acute Survival Studies2007https://link.springer.com/content/pdf/10.1007/0-387-49416-2_15.pdf71-75The primary use of venous catheterization is for intravenous administration. This route is advantageous due to the predictability and reproducibility of drug kinetics. One is able to deliver agents in a controlled manner via pumps. Intravenous deliveries bypass the stomach and, therefore, avoid gastric pH and first pass metabolism issues. There is rapid onset at the target site. This is the route of choice for protein and peptide-based drugs. There are several considerations to note prior to intravenous delivery. One must be aware of possible vascular irritation and damage caused by the test substance, leakage out of the vessel, volume overload (hemodilution), hemolysis, and infection. One must also carefully consider the solubility and pH of the test substance. Solution should be 7.4 pH adjusted and completely soluble. Any particulates will cause an emboli. Methods in vascular infusion biotechnology in research with rodents by Dr. Nolan (2002) is an excellent resource for this applicationBioanalytical Systems, Inc. (BAS) 2701 Kent Avenue West Lafayette, Indiana 47906 www.bioanalytical.com 72 Page 3. Use of arterial catheterization Bioanalytical Systems, Inc. (Culex) 2701 Kent Avenue West Lafayette, Indiana 47906 www.bioanalytical.com DiLab Inc2007-06-01 00:00:0010.1007/0-387-49416-2_15
Recent progress in in-vivo sampling and analysisMusteata, F;MicrosamplingData ReviewTrAC Trends in Analytical Chemistry2013https://www.sciencedirect.com/science/article/pii/S0165993613000186154-16845This article reviews recent developments in in-vivo sampling and sample preparation for analysis of biological systems. The trend towards microanalytical techniques is justified by the small amount of sample that is available from some biological systems and the need to minimize interference with the system that is being studied or analyzed. Different approaches to direct in-vivo sampling and sample preparation are described, including microsampling, ultrafiltration, microextraction, microdialysis, solid-phase microextraction, biosensors, ambient mass spectrometry, spectroscopy, and microfluidic devices.8.428One such notable example is the Culex automated sampling system developed by BASi to collect analytical instruments can be performed either manually, using laboratory robotics (eg, Culex), or by Left: Picture of a conscious rat with the headpiece mounted on his head during 2013-04-01 00:00:0010.1016/j.trac.2012.11.012
Safety Pharmacology Society: 9th Annual MeetingCavero, I;Safety PharmacologyData ReviewExpert Opin Drug Saf2010https://www.tandfonline.com/doi/abs/10.1517/14740331003625577365-3789220175703The keynote presentation of the Safety Pharmacology (SP) Society 9th Annual Meeting addressed the urgency, for pharmaceutical organizations, to implement strategies for effectively communicating drug risks to all concerned stakeholders and, in particular, the general public. The application of chronobiology to SP investigational protocols can improve the search of drug-induced adverse effects. The Distinguished Service Award Lecture reviewed a life-long journey through trials and tribulations in the quest of the ever-distant scientific truth. The revision process of Directive 86/609/EC for improving animal welfare should be conducted with the purpose of maintaining a fair balance among animal protection, human health and research imperatives in order to prevent the migration of pharmaceutical activities outside Europe. Additional topics of interest were the behavioral, metabolic and cardiovascular problems experienced by small animals housed at the standard laboratory temperature. A technology for the automated collection of blood and urine samples in rats implanted with telemetry sensors was presented. Non-clinical, clinical, regulatory and legal aspects of abuse liability were expertly reviewed. The 'degradability' of pharmaceuticals into environment-friendly chemicals should be an actively searched and optimized feature of future pharmaceuticals in order to prevent drug pollution of ecosystems. Transgenic and diseased animal models should be selected whenever they can facilitate the determination of drug-induced adverse effects. SP strategies to investigate the safety of drug combination products were exemplified and analyzed in depth. The future of SP was proposed to lie not in the performance of regulatory studies of pharmacodynamic nature but in developing and early applying an array of screening assays for clearing clinical candidates against known drug-induced organ function injuries. In conclusion, the 2009 SP Society annual meeting offered a wealth of thought-provoking material to attendees for improving SP investigation strategies.3.22Germany); 'Effects of body temperature on QT interval in beagle dog' (Dr Phillip Atterson This biological feature obliges the mouse to a relentless search for food to is obtained by combining custom - modified, commercially available equipment (automated BASi Culex- ABS O2010-03-01 00:00:0010.1517/14740331003625577
Safety pharmacology--current and emerging conceptsHamdam, J;Sethu, S;Smith, T;Alfirevic, A;Alhaidari, M;Atkinson, J;Ayala, M;Box, H;Cross, M;Delaunois, A;Dermody, A;Govindappa, K;Guillon, JM;Jenkins, R;Kenna, G;Lemmer, B;Meecham, K;Olayanju, A;Pestel, S;Rothfuss, A;Sidaway, J;Sison-Young, R;Smith, E;Stebbings, R;Tingle, Y;Valentin, JP;Williams, A;Williams, D;Park, K;Goldring, C;Safety PharmacologyData ReviewToxicol. Appl. Pharmacol.2013https://www.sciencedirect.com/science/article/pii/S0041008X13002548229-241273223732082Safety pharmacology (SP) is an essential part of the drug development process that aims to identify and predict adverse effects prior to clinical trials. SP studies are described in the International Conference on Harmonisation (ICH) S7A and S7B guidelines. The core battery and supplemental SP studies evaluate effects of a new chemical entity (NCE) at both anticipated therapeutic and supra-therapeutic exposures on major organ systems, including cardiovascular, central nervous, respiratory, renal and gastrointestinal. This review outlines the current practices and emerging concepts in SP studies including frontloading, parallel assessment of core battery studies, use of non-standard species, biomarkers, and combining toxicology and SP assessments. Integration of the newer approaches to routine SP studies may significantly enhance the scope of SP by refining and providing mechanistic insight to potential adverse effects associated with test compounds. Copyright © 2013 Elsevier Inc. All rights reserved.3.585However, an integrated pharmacology testing system in surgically prepared rats has been recently developed for simultaneous measurements of GFR and renal plasma flow. This system successfully combined BASi Culex® automated blood sampling, radiotelemetry, quantitative urinalysis, and nephron site-specific urinary biomarkers of injury into one model testing system (Kamendi et al., 2010, Litwin et al., 2011). Renal toxicity can be predicted using clinical chemistry following a single administration of the test drug (Pestel et al., 2006), but the sensitivity is rather low when compared to NMR-based metabolomics methods (Lienemann et al., 2008)2013-12-01 00:00:0010.1016/j.taap.2013.04.039
Samidorphan mitigates olanzapine-induced weight gain and metabolic dysfunction in rats and non-human primatesCunningham, JI;Eyerman, DJ;Todtenkopf, MS;Dean, RL;Deaver, DR;Sanchez, C;Namchuk, M;Pharmacokinetics (PK)AntipsychoticJ. Psychopharmacol. (Oxford)2019https://journals.sagepub.com/doi/abs/10.1177/0269881119856850269881119856850331031294646Olanzapine, regarded as one of the most efficacious antipsychotic medications for the treatment of schizophrenia, is associated with a high risk of weight gain and metabolic dysfunction. ALKS 3831, a clinical candidate for treatment of schizophrenia, is a combination of olanzapine and samidorphan, an opioid receptor antagonist. The addition of samidorphan is intended to mitigate weight gain and the metabolic dysregulation associated with the use of olanzapine. Non-clinical studies were conducted to assess the metabolic effects of olanzapine and samidorphan alone and in combination at clinically relevant exposure levels. Chronic olanzapine administration in male and female rats shifted body composition by increasing adipose mass, which was accompanied by an increase in the rate of weight gain in female rats. Co-administration of samidorphan normalized body composition in both sexes and attenuated weight gain in female rats. In hyperinsulinemic euglycemic clamp experiments conducted prior to measurable changes in weight and/or body composition, olanzapine decreased hepatic insulin sensitivity and glucose uptake in muscle while increasing uptake in adipose tissue. Samidorphan appeared to normalize glucose utilization in both tissues, but did not restore hepatic insulin sensitivity. In subsequent studies, samidorphan normalized olanzapine-induced decreases in whole-body glucose clearance following bolus insulin administration. Results from experiments in female monkeys paralleled the effects in rats. Olanzapine administration increased weight gain and adiposity, both of which were attenuated by samidorphan. Furthermore, the combination of olanzapine and samidorphan prevented olanzapine-induced insulin insensitivity. Collectively, these data indicate that samidorphan mitigates several metabolic abnormalities associated with olanzapine in both the presence and the absence of weight gain.4.221d with 0.375 U/kg of insulin for all studies. Rats were treated with OLZ, SAM or the combination 48 h prior to insulin sensitivity testing. Rats were placed in an automated blood sampling system (CULEX; BASi, West Lafayette, Indiana, USA) and fasted overnight for 16–18 h prior to test. The following morning, baseline glucose concentrations were measured in whole blood using a glucometer (N;Olanzapine, regarded as one of the most efficacious antipsychotic medications for the treatment of schizophrenia, is associated with a high risk of weight gain and metabolic dysfunction. ALKS 3831, a clinical candidate for treatment of schizophrenia, is a combination of olanzapine and samidorphan, an opioid receptor antagonist. The addition of samidorphan is intended to mitigate weight gain and the metabolic dysregulation associated with the use of olanzapine. Non-clinical studies were conducted to assess the metabolic effects of olanzapine and samidorphan alone and in combination at clinically relevant exposure levels. Chronic olanzapine administration in male and female rats shifted body composition by increasing adipose mass, which was accompanied by an increase in the rate of weight gain in female rats. Co-administration of samidorphan normalized body composition in both sexes and attenuated weight gain in female rats. In hyperinsulinemic euglycemic clamp experiments conducted prior to measurable changes in weight and/or body composition, olanzapine decreased hepatic insulin sensitivity and glucose uptake in muscle while increasing uptake in adipose tissue. Samidorphan appeared to normalize glucose utilization in both tissues, but did not restore hepatic insulin sensitivity. In subsequent studies, samidorphan normalized olanzapine-induced decreases in whole-body glucose clearance following bolus insulin administration. Results from experiments in female monkeys paralleled the effects in rats. Olanzapine administration increased weight gain and adiposity, both of which were attenuated by samidorphan. Furthermore, the combination of olanzapine and samidorphan prevented olanzapine-induced insulin insensitivity. Collectively, these data indicate that samidorphan mitigates several metabolic abnormalities associated with olanzapine in both the presence and the absence of weight gain2019-07-11 00:00:0010.1177/0269881119856850
Saturation of multidrug-resistant protein 2 (mrp2/abcc2)-mediated hepatobiliary secretion: nonlinear pharmacokinetics of a heterocyclic compound in rats after intravenous bolus administrationHu, Y;Sampson, KE;Heyde, BR;Mandrell, KM;Li, N;Zutshi, A;Lai, Y;ADME (Bile)Drug MetabolismDrug Metab. Dispos.2009http://dmd.aspetjournals.org/content/37/4/841.short841-84637419139164Multidrug-resistant protein 2 (MRP2/ABCC2), expressed on the canalicular membrane of hepatocytes, mediates the secretion of conjugated or nonconjugated compounds into bile and plays an important role in physiology and drug elimination. A heterocyclic compound, BPCPU [1-(1-(4-bromophenyl)-3-carbamoyl-1H-pyrazol-4-yl) urea], which was metabolically stable in vitro in rat liver microsomes and freshly isolated rat hepatocytes, demonstrated a saturable nonlinear pharmacokinetic profile in the rat. Polarized efflux was observed for this compound in Caco-2 cells, with a low K(m) = 1.06 +/- 0.06 microM. The Caco-2 efflux was dose-dependent and saturable. Coadministration of 25 microM MK571 ([3-[[3-[2-(7-chloroquinolin-2-yl)vinyl]phenyl]-(2-dimethylcarbamoylethylsulfanyl)methylsulfanyl] propionic acid]), an MRP inhibitor, blocked the polarized efflux in Caco-2 cells. In contrast, this compound did not inhibit calcein efflux in MRP2 gene-transfected Madin-Darby canine kidney cells, suggesting that it is a substrate, not an inhibitor, of the MRP2/ABCC2 transporter. To investigate the mechanism for the nonlinear pharmacokinetics, bile duct-cannulated rats were used to obtain time profiles of plasma concentration, biliary, and urinary excretion after intravenous administration at various doses. The plasma clearance increased remarkably with decreased dose, from 1.5 ml/min/kg at 5 mg/kg to 14.9 ml/min/kg at 0.05 mg/kg. A dose-dependent biliary excretion also was observed. The results revealed that saturation of hepatobiliary secretion played a role in the dose-dependent changes in total body clearance and biliary clearance. Saturating concentrations of the Mrp2/Abcc2 substrate, BPCPU, causing decreased hepatobiliary clearance could be the major cause for the nonlinear pharmacokinetics observed in rats.3.354The rats were surgically implanted with BASi vascular catheters (BAS Bioanalytical Systems, West Lafayette, IN) in both the carotid artery and jugular vein. The common bile duct was also catheterized with the cannula exteriorized for access between the scapulae. Animals were acclimated in Culex cages (BASi) overnight before dosing with food and water provided ad libitum. Animals were dosed with the BPCPU through the jugular vein catheter. The dosing volume was controlled at approximately 300 μl (1 μl/g body weight). Blood samples were automatically collected by the Culex at designated intervals to 24 h postdose. Bile samples were collected in 1-h increments up to 8 h and then continuously from 8 to 24 h, after a baseline predose collection. All procedures were approved by the St. Louis Pfizer Institutional Animal Care and Use Committee. The animal care and use program is fully accredited by the Association for Assessment and Accreditation of Laboratory Animal Care, International2009-04-01 00:00:0010.1124/dmd.108.024059
Session IV-4: Multi-imaging modalities, telemetry and the Three RsCroy, BA;Adams, MA;Safety PharmacologyPre-eclampsiapdfs.semanticscholar.org2011https://pdfs.semanticscholar.org/fca7/c47cec70f65748583814ff8828ab1907384f.pdf1-4Mice are valuable tools providing understanding of the physiology and pathologies of human pregnancy. Acute onset hypertension with kidney failure is the commonest (3-7%) human pregnancy complication, characterizing the syndrome pre-eclampsia, a medical emergency with immunological complications. We implanted PA-C10 radiotransmitters (DSI) into 8 week female mice. After 10 days recovery, instrumented mice were paired for mating. Upon copulation plug detection, data were collected continuously to 48 h postpartum. A fluctuating pattern of normal blood pressure was defined. The pattern, observed in randombred CD1, inbred C57BL/6J, BALB/cJ, normoglycemic NOD and immune deficient Rag2-/- and Rag2-/-/Il2rg-/-, changed at specific times important for placental development. NOD scid and hyperglycemic NOD pregnancies differed, identifying NK cells and blood glucose values respectively as factors contributing to circulatory control over pregnancy. The precision of telemetric measurement and the concordance of findings between replicate animals made 4-6 recordings sufficient to generate publication quality data. Complications and technical failures were common in these pregnancy-based studies, requiring preparation of 10-12 instrumented mice per group. These animal numbers are much lower than needed for pregnancy time course studies requiring daily euthanasia of 3-6 replicate mice. Previous attempts to collect this information using serial, daily, tail cuff recording gave data with sufficient variability that we concluded (incorrectly) mice differed from humans because they lacked a dynamic pattern of cardiovascular regulation across pregnancy.in rats. For this, a unique engineering solution was created to combine the BASi Culex® automated blood sampling system with Data Sciences International Physio-tel® Multiplus radio telemetry system. Studies were performed2011-01-01 00:00:00
Small molecular inhibitor of transforming growth factor-beta protects against development of radiation-induced lung injuryAnscher, MS;Thrasher, B;Zgonjanin, L;Rabbani, ZN;Corbley, MJ;Fu, K;Sun, L;Lee, WC;Ling, LE;Vujaskovic, Z;Pharmacokinetics (PK)Lung InjuryInt. J. Radiat. Oncol. Biol. Phys.2008https://www.sciencedirect.com/science/article/pii/S0360301608003416829-83771318411002To determine whether an anti-transforming growth factor-beta (TGF-beta) type 1 receptor inhibitor (SM16) can prevent radiation-induced lung injury. One fraction of 28 Gy or sham radiotherapy (RT) was administered to the right hemithorax of Sprague-Dawley rats. SM16 was administered in the rat chow (0.07 g/kg or 0.15 g/kg) beginning 7 days before RT. The rats were divided into eight groups: group 1, control chow; group 2, SM16, 0.07 g/kg; group 3, SM16, 0.15 g/kg; group 4, RT plus control chow; group 5, RT plus SM16, 0.07 g/kg; group 6, RT plus SM16, 0.15 g/kg; group 7, RT plus 3 weeks of SM16 0.07 g/kg followed by control chow; and group 8, RT plus 3 weeks of SM16 0.15 g/kg followed by control chow. The breathing frequencies, presence of inflammation/fibrosis, activation of macrophages, and expression/activation of TGF-beta were assessed. The breathing frequencies in the RT plus SM16 0.15 g/kg were significantly lower than the RT plus control chow from Weeks 10-22 (p <0.05). The breathing frequencies in the RT plus SM16 0.07 g/kg group were significantly lower only at Weeks 10, 14, and 20. At 26 weeks after RT, the RT plus SM16 0.15 g/kg group experienced a significant decrease in lung fibrosis (p = 0.016), inflammatory response (p = 0.006), and TGF-beta1 activity (p = 0.011). No significant reduction was found in these measures of lung injury in the group that received SM16 0.7 g/kg nor for the short-course (3 weeks) SM16 at either dose level. SM16 at a dose of 0.15 g/kg reduced functional lung damage, morphologic changes, inflammatory response, and activation of TGF-beta at 26 weeks after RT. The data suggest a dose response and also suggest the superiority of long-term vs. short-term dosing.6.203A 430MW inhibitor of TGF-βRI kinase, SM16 14, 15, was formulated into rat chow (Research Diets, New Brunswick, NJ 0.07 g/kg SM16 chow ad libitum for 4 days, and then placed in a Culex Automated Pharmacology System (Bioanalytical Systems, Lafayette, IN2008-07-01 00:00:0010.1016/j.ijrobp.2008.02.046
Somatostatin is essential for the sexual dimorphism of GH secretion, corticosteroid-binding globulin production, and corticosterone levels in miceAdams, JM;Otero-Corchon, V;Hammond, GL;Veldhuis, JD;Qi, N;Low, MJ;Hormone ResearchEndocrinologyEndocrinology2015https://academic.oup.com/endo/article-abstract/156/3/1052/24231771052-1065156325551181Distinct male and female patterns of pituitary GH secretion produce sexually differentiated hepatic gene expression profiles, thereby influencing steroid and xenobiotic metabolism. We used a fully automated system to obtain serial nocturnal blood samples every 15 minutes from cannulated wild-type (WT) and somatostatin knockout (Sst-KO) mice to determine the role of SST, the principal inhibitor of GH release, in the generation of sexually dimorphic GH pulsatility. WT males had lower mean and median GH values, less random GH secretory bursts, and longer trough periods between GH pulses than WT females. Each of these parameters was feminized in male Sst-KO mice, whereas female Sst-KO mice had higher GH levels than all other groups, but GH pulsatility was unaffected. We next performed hepatic mRNA profiling with high-density microarrays. Male Sst-KO mice exhibited a globally feminized pattern of GH-dependent mRNA levels, but female Sst-KO mice were largely unaffected. Among the differentially expressed female-predominant genes was Serpina6, which encodes corticosteroid-binding globulin (CBG). Increased CBG was associated with elevated diurnal peak plasma corticosterone in unstressed WT females and both sexes of Sst-KO mice compared with WT males. Sst-KO mice also had exaggerated ACTH and corticosterone responses to acute restraint stress. However, consistent with their lack of phenotypic signs of excess glucocorticoids, cerebrospinal fluid concentrations of free corticosterone in Sst-KO mice were not elevated. In summary, SST is necessary for the prolonged interpulse troughs that define masculinized pituitary GH secretion. SST also contributes to sexual dimorphism of the hypothalamic-pituitary-adrenal axis via GH-dependent regulation of hepatic CBG production.3.8al activity and body weight regain to more than or equal to 90% of their presurgery level. They were acclimatized for 3–4 days to Ratturn cages as part of the Culex automated blood sampling system (Basi, Inc), and then, 44 serial blood samples were obtained from the carotid cannulae at 15-minute intervals during the 12-hour nocturnal period in complete darkness starting at 7 pm. Because of the d ... rawn at each time point, 10 μL was diluted with 50 μL of heparinized saline into refrigerated glass tubes, and the remaining 45 μL blood with 10 μL of heparinized saline was infused back into the mouse to maintain blood volume. The next morning, all mice appeared healthy. Collection tubes were centrifuged, and 50 μL of clear, diluted plasma was frozen at −20°C before assay. Samples were co ... measurements less than the lowest point of the standard curve were assigned that value (7.8 ng/mL). Basal daytime corticosterone measurements were measured by an in-house assay at the Oregon National Primate Research Center hormone assay core as previously described (23). Plasma ACTH was measured by immunoradiometric assay (Nichols Institute). For the restraint stress and CSF collection experiment ... They found elevated plasma IGF-I, increased pituitary _Gh_, _Ghrhr_, and _Ghsr_ expression, and decreased hypothalamic _Ghrh_ expression only in female _Sst_-KO mice, leading them to conclude that endogenous SST plays a predominant role in females in the regulation of the GH axis (67). Those findings are consistent with our observations that female _Sst_-KO mice exhibited the highest levels of me;... al activity and body weight regain to more than or equal to 90% of their presurgery level. They were acclimatized for 3–4 days to Ratturn cages as part of the Culex automated blood sampling system (Basi, Inc), and then, 44 serial blood samples were obtained from the carotid cannulae at 15-minute intervals during the 12-hour nocturnal period in complete darkness starting at 7 pm. Because of the d ... rawn at each time point, 10 μL was diluted with 50 μL of heparinized saline into refrigerated glass tubes, and the remaining 45 μL blood with 10 μL of heparinized saline was infused back into the mouse to maintain blood volume. The next morning, all mice appeared healthy. Collection tubes were centrifuged, and 50 μL of clear, diluted plasma was frozen at −20°C before assay. Samples were co ... measurements less than the lowest point of the standard curve were assigned that value (7.8 ng/mL). Basal daytime corticosterone measurements were measured by an in-house assay at the Oregon National Primate Research Center hormone assay core as previously described (23). Plasma ACTH was measured by immunoradiometric assay (Nichols Institute). For the restraint stress and CSF collection experiment ... They found elevated plasma IGF-I, increased pituitary _Gh_, _Ghrhr_, and _Ghsr_ expression, and decreased hypothalamic _Ghrh_ expression only in female _Sst_-KO mice, leading them to conclude that endogenous SST plays a predominant role in females in the regulation of the GH axis (67). Those findings are consistent with our observations that female _Sst_-KO mice exhibited the highest levels of me;... al activity and body weight regain to more than or equal to 90% of their presurgery level. They were acclimatized for 3–4 days to Ratturn cages as part of the Culex automated blood sampling system (Basi, Inc), and then, 44 serial blood samples were obtained from the carotid cannulae at 15-minute intervals during the 12-hour nocturnal period in complete darkness starting at 7 pm. Because of the d ... rawn at each time point, 10 μL was diluted with 50 μL of heparinized saline into refrigerated glass tubes, and the remaining 45 μL blood with 10 μL of heparinized saline was infused back into the mouse to maintain blood volume. The next morning, all mice appeared healthy. Collection tubes were centrifuged, and 50 μL of clear, diluted plasma was frozen at −20°C before assay. Samples were co ... measurements less than the lowest point of the standard curve were assigned that value (7.8 ng/mL). Basal daytime corticosterone measurements were measured by an in-house assay at the Oregon National Primate Research Center hormone assay core as previously described (23). Plasma ACTH was measured by immunoradiometric assay (Nichols Institute). For the restraint stress and CSF collection experiment ... They found elevated plasma IGF-I, increased pituitary _Gh_, _Ghrhr_, and _Ghsr_ expression, and decreased hypothalamic _Ghrh_ expression only in female _Sst_-KO mice, leading them to conclude that endogenous SST plays a predominant role in females in the regulation of the GH axis (67). Those findings are consistent with our observations that female _Sst_-KO mice exhibited the highest levels of me;... al activity and body weight regain to more than or equal to 90% of their presurgery level. They were acclimatized for 3–4 days to Ratturn cages as part of the Culex automated blood sampling system (Basi, Inc), and then, 44 serial blood samples were obtained from the carotid cannulae at 15-minute intervals during the 12-hour nocturnal period in complete darkness starting at 7 pm. Because of the d;... al activity and body weight regain to more than or equal to 90% of their presurgery level. They were acclimatized for 3–4 days to Ratturn cages as part of the Culex automated blood sampling system (Basi, Inc), and then, 44 serial blood samples were obtained from the carotid cannulae at 15-minute intervals during the 12-hour nocturnal period in complete darkness starting at 7 pm. Because of the d ... rawn at each time point, 10 μL was diluted with 50 μL of heparinized saline into refrigerated glass tubes, and the remaining 45 μL blood with 10 μL of heparinized saline was infused back into the mouse to maintain blood volume. The next morning, all mice appeared healthy. Collection tubes were centrifuged, and 50 μL of clear, diluted plasma was frozen at −20°C before assay. Samples were co ... measurements less than the lowest point of the standard curve were assigned that value (7.8 ng/mL). Basal daytime corticosterone measurements were measured by an in-house assay at the Oregon National Primate Research Center hormone assay core as previously described (23). Plasma ACTH was measured by immunoradiometric assay (Nichols Institute). For the restraint stress and CSF collection experiment ... They found elevated plasma IGF-I, increased pituitary _Gh_, _Ghrhr_, and _Ghsr_ expression, and decreased hypothalamic _Ghrh_ expression only in female _Sst_-KO mice, leading them to conclude that endogenous SST plays a predominant role in females in the regulation of the GH axis (67). Those findings are consistent with our observations that female _Sst_-KO mice exhibited the highest levels of me2015-03-01 00:00:0010.1210/en.2014-1429
Suppression and resolution of autoimmune arthritis by rhesus θ-defensin-1, an immunomodulatory macrocyclic peptideSchaal, JB;Tran, DQ;Subramanian, A;Patel, R;Laragione, T;Roberts, KD;Trinh, K;Tongaonkar, P;Tran, PA;Minond, D;Fields, GB;Beringer, P;Ouellette, AJ;Gulko, PS;Selsted, ME;Pharmacokinetics (PK)Macrocyclic PeptidesPLoS ONE2017https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0187868&_ga=2.176116900.74143535.1533772800-2020293321.1533772800e0187868121129145473θ-defensins constitute a family of macrocyclic peptides expressed exclusively in Old World monkeys. The peptides are pleiotropic effectors of innate immunity, possessing broad spectrum antimicrobial activities and immunoregulatory properties. Here we report that rhesus θ-defensin 1 (RTD-1) is highly effective in arresting and reversing joint disease in a rodent model of rheumatoid arthritis (RA). Parenteral RTD-1 treatment of DA/OlaHsd rats with established pristane-induced arthritis (PIA) rapidly suppressed joint disease progression, restored limb mobility, and preserved normal joint architecture. RTD-1 significantly reduced joint IL-1β levels compared with controls. RTD-1 dose-dependently inhibited fibroblast-like synoviocyte (FLS) invasiveness and FLS IL-6 production. Consistent with the inhibition of FLS invasiveness, RTD-1 was a potent inhibitor of arthritogenic proteases including ADAMs 17 and 10 which activate TNFα, and inhibited matrix metalloproteases, and cathepsin K. RTD-1 was non-toxic, non-immunogenic, and effective when administered as infrequently as once every five days. Thus θ-defensins, which are absent in humans, have potential as retroevolutionary biologics for the treatment of RA.2.776Sequential blood sampling for pharmacokinetic analyses was performed by Bioanalytical Systems, Inc were fitted with a surgically implanted jugular vein catheter for sequential blood collection (Culex NxT Automated In Vivo Sampling System; BASi, West Lafayette2017-11-16 00:00:0010.1371/journal.pone.0187868
Synthesis and biological evaluation of polysulfated oligosaccharide glycosides as inhibitors of angiogenesis and tumor growthJohnstone, KD;Karoli, T;Liu, L;Dredge, K;Copeman, E;Li, CP;Davis, K;Hammond, E;Bytheway, I;Kostewicz, E;Chiu, FC;Shackleford, DM;Charman, SA;Charman, WN;Harenberg, J;Gonda, TJ;Ferro, V;Pharmacokinetics (PK)Cancer/OncologyJ. Med. Chem.2010https://pubs.acs.org/doi/abs/10.1021/jm901449m1686-169953420128596A series of polysulfated penta- and tetrasaccharide glycosides containing alpha(1-->3)/alpha(1-->2)-linked mannose residues were synthesized as heparan sulfate (HS) mimetics and evaluated for their ability to inhibit angiogenesis. The compounds bound tightly to angiogenic growth factors (FGF-1, FGF-2, and VEGF) and strongly inhibited heparanase activity. In addition, the compounds exhibited potent activity in cell-based and ex vivo assays indicative of angiogenesis, with tetrasaccharides exhibiting activity comparable to that of pentasaccharides. Selected compounds also showed good antitumor activity in vivo in a mouse melanoma (solid tumor) model resistant to the phase III HS mimetic 1 (muparfostat, formerly known as PI-88). The lipophilic modifications also resulted in reduced anticoagulant activity, a common side effect of HS mimetics, and conferred a reasonable pharmacokinetic profile in the rat, as exemplified by the sulfated octyl tetrasaccharide 5. The data support the further investigation of this class of compounds as potential antiangiogenic, anticancer therapeutics.6.054Compound 5, formulated as a 2.8 mg/mL solution in PBS, was infused intravenously over a period of 5 min (1.0 mL/rat; n = 2 rats) or injected subcutaneously between the scapulae (1.0 mL/rat; n = 2 rats) to achieve a nominal dose of 10 mg/kg. Samples of blood [collected from the arterial cannula using a Culex autosampler (BASi)] and total urine were collected periodically for up to 8 and 24 h postdose, respectively. To minimize the potential for ex vivo degradation of 5 in blood/plasma samples, blood samples were collected directly into heparinized borosilicate vials (at 4 C) containing Complete (a protease inhibitor cocktail, 80 mg/mL, Roche, Mannheim, Germany), potassium fluoride (4 mg/mL), and EDTA (0.1 M). Once collected, blood samples were centrifuged, and the plasma supernatant was removed and stored at -20 C for subsequent analysis by LC-MS2010-02-25 00:00:0010.1021/jm901449m
Synthesis, structure-activity relationships and brain uptake of a novel series of benzopyran inhibitors of insulin-regulated aminopeptidaseMountford, SJ;Albiston, AL;Charman, WN;Ng, L;Holien, JK;Parker, MW;Nicolazzo, JA;Thompson, PE;Chai, SY;Pharmacokinetics (PK)MemoryJ. Med. Chem.2014https://pubs.acs.org/doi/abs/10.1021/jm401540f1368-137757424471437Peptide inhibitors of insulin-regulated aminopeptidase (IRAP) enhance fear avoidance and spatial memory and accelerate spatial learning in a number of memory paradigms. Using a virtual screening approach, a series of benzopyran compounds was identified that inhibited the catalytic activity of IRAP, ultimately resulting in the identification of potent and specific inhibitors. The present study describes the medicinal chemistry campaign that led to the development of the lead candidate, 3, highlighting the key structural features considered as critical for binding. Furthermore, the in vivo pharmacokinetics and brain uptake of compounds (1 and 3) were assessed in rats and were complemented with in vitro human and rat microsomal stability studies. Following intravenous administration to rodents, 3 exhibits brain exposure, albeit it is rapidly converted to 1, a compound which also exhibits potent inhibition of IRAP.6.054All animal studies were performed in accordance with the Australian National Health and Medical Research Council code of practice for the care and use of animals for scientific purposes and were approved by the Monash University (Victorian College of Pharmacy) Animal Ethics committee. Male Sprague−Dawley rats (7−9 weeks) were intravenously administered (via an indwelling jugular vein cannula) a 1.0 mL solution of 3 (2 mg/kg in an aqueous solution containing 30% w/v hydroxypropyl-β-cyclodextrin and 1% v/v DMSO) over a 5 min period. In additional studies, rats were intraperitoneally administered a 0.5 mL solution of 3 (10 mg/kg in in 10% DMSO (v/v)) and 30% hydroxypropyl-β-cyclodextrin (w/v). Samples of arterial blood were collected (from an indwelling carotid artery cannula) over 24 h using a Culex automated blood sampler (BASi Instruments, West LaFayette, IN, USA). Blood samples were collected directly into heparinized borosilicate vials (at 4 °C) containing Complete protease inhibitor cocktail (80 mg/mL, Roche, Mannheim, Germany), EDTA (0.1 M), and potassium fluoride (4 mg/mL) (stabilization mixture). Blood samples were immediately centrifuged and the plasma supernatant separated and immediately stored at −20 °C to minimize potential ex vivo conversion of 3 to 1. All plasma samples were subsequently assayed for 3 and 1 concentration using a validated LCMS method, with a lower limit of quantification of 1 ng/mL for 3 and 0.5 ng/mL for 1. Noncompartmental pharmacokinetic analysis was performed using WinNonLin software (version 4.0, Pharsight Corporation, Mountain View, CA, USA) to estimate the terminal elimination halflife, total plasma clearance, and volume of distribution2014-02-27 00:00:0010.1021/jm401540f
The contribution of VEGF signalling to fostamatinib-induced blood pressure elevationSkinner, M;Philp, K;Lengel, D;Coverley, L;Lamm Bergström, E;Glaves, P;Musgrove, H;Prior, H;Braddock, M;Huby, R;Curwen, JO;Duffy, P;Harmer, AR;Safety PharmacologyBlood PressureBr. J. Pharmacol.2014https://bpspubs.onlinelibrary.wiley.com/doi/abs/10.1111/bph.125592308-2320171924329544Fostamatinib is an inhibitor of spleen tyrosine kinase (TK). In patients, fostamatinib treatment was associated with increased BP. Some TK inhibitors cause BP elevation, by inhibiting the VEGF receptor 2 (VEGFR2). Here, we have assessed the mechanistic link between fostamatinib-induced BP elevation and inhibition of VEGF signalling. We used conscious rats with automated blood sampling and radio telemetry and anaesthetized rats to measure cardiovascular changes. Rat isolated aorta and isolated hearts, and human resistance vessels in vitro were also used. NO production by human microvascular endothelial cells was measured with the NO-dependent probe, DAF-FM and VEGFR2 phosphorylation was determined in mouse lung, ex vivo. In conscious rats, fostamatinib dose-dependently increased BP. The time course of the BP effect correlated closely with the plasma concentrations of R406 (the active metabolite of fostamatinib). In anaesthetized rats, infusion of R406 increased BP and decreased femoral arterial conductance. Endothelial function was unaffected, as infusion of R406 did not inhibit hyperaemia- or ACh-induced vasodilatation in rats. R406 did not affect contraction of isolated blood vessels. R406 inhibited VEGF-stimulated NO production from human endothelial cells in vitro, and treatment with R406 inhibited VEGFR2 phosphorylation in vivo. R406 inhibited VEGF-induced hypotension in anaesthetized rats. Increased vascular resistance, secondary to reduced VEGF-induced NO release from endothelium, may contribute to BP increases observed with fostamatanib. This is consistent with the elevated BP induced by other drugs inhibiting VEGF signalling, although the contribution of other mechanisms cannot be excluded. © 2013 The British Pharmacological Society.6.583to the AstraZeneca facility and, after the acclimatisation period, the femoral catheters were connected to a Culex® automated blood sampler (BASi, West Lafayette Where free plasma concentrations are quoted, this is based on plasma protein binding in the rat of 97.92014-05-01 00:00:0010.1111/bph.12559
The Effects of Arginine Vasopressin and Acetazolamide on CNS Clearance of Acetaminophen and Ibuprofen in RatsWang, Q;Grater, R;LuisetteDelva, EV;Black, C;PK/PDCNSJ Neurol Neuromedicine2018https://www.jneurology.com/articles/the-effects-of-arginine-vasopressin-and-acetazolamide-on-cns-clearance-of-acetaminophen-and-ibuprofen-in-rats.pdf51-56day 1, the animals were individually placed into Culex cages (BASi, West Lafayette On day 1, animals were individually placed into the Culex auto-sampler cages Monoclonal antibody exposure in rat and cynomolgus monkey cerebrospinal fluid following systemic administration2018-01-01 00:00:00
The effects of environmental enrichment on the welfare of laboratory swine housed in isolationDeBoer, SP;Stress HormonesBehaviorThesis2012http://search.proquest.com/openview/0a54e9134d53ad7150eda0d78246ab40/1?pq-origsite=gscholar&cbl=18750&diss=yPage 1. Graduate School ETD Form 9 (Revised 12/07) PURDUE UNIVERSITY GRADUATE SCHOOL Thesis/Dissertation Acceptance This is to certify that the thesis/dissertation prepared By Entitled For the degree of Is approved by the final examining committee: Chair ;l movement or slippage while on the pig. Automated blood collection was controlled by an automated sampling system (Culex, BioAnalytical Systems Inc., West Lafayette, IN, USA). The sampling system (Culex) was programmed to collect one 1 mL blood sample 2 hours after the program was started, timer beginning immediately after the person responsible for husbandry le2012-01-01 00:00:00
The evaluation of radiolabeled artesunate on tissue distribution in rats and protein binding in humansLi, Q;Xie, LH;Haeberle, A;Zhang, J;Weina, P;Pharmacokinetics (PK)AntimalarialAm. J. Trop. Med. Hyg.2006https://www.ajtmh.org/content/journals/10.4269/ajtmh.2006.75.817817-82675517123971The present study reports the tissue distribution, pharmacokinetics, mass balance, and elimination of [(14)C] artesunate (AS) following single intravenous administration in rats. Protein binding was performed with rat and human plasma. Radioactivity and drug levels in blood, plasma, tissues, urine, and feces up to 192 hours were collected and measured. The mean terminal half-life of plasma (76 h) and blood (105 h) radioactivity was prolonged compared with that of unchanged AS (0.43 h) and dihydroartemisinin (0.75 h), an active metabolite of AS. Drug was widely distributed after 1 hour in select tissues. After 24 hours, the radioactivity rapidly declined in all tissues except spleen until 96 hours. Only 1% of total radioactivity was detected in brain tissue. AS revealed a higher binding capacity with human and rat plasma proteins (73-81%). The radioactivity in whole blood was higher (two to fourfold) than that in plasma throughout the period of the treatment, suggesting that AS binding to RBCs may relate to its powerful antimalarial activity.2.315The Culex auto- mated blood sampler provides a way for blood withdrawal at preprogrammed intervals Rat samples were freshly collected from 5 male and 5 female rats with reductive electrochemi- cal detection (HPLC-ECD) was performed utilizing a model BAS 200B liquid2006-11-01 00:00:0010.4269/ajtmh.2006.75.817
The hypoxia imaging agent CuII(atsm) is neuroprotective and improves motor and cognitive functions in multiple animal models of Parkinson's diseaseHung, LW;Villemagne, VL;Cheng, L;Sherratt, NA;Ayton, S;White, AR;Crouch, PJ;Lim, S;Leong, SL;Wilkins, S;George, J;Roberts, BR;Pham, CL;Liu, X;Chiu, FC;Shackleford, DM;Powell, AK;Masters, CL;Bush, AI;O'Keefe, G;Culvenor, JG;Cappai, R;Cherny, RA;Donnelly, PS;Hill, AF;Finkelstein, DI;Barnham, KJ;Pharmacokinetics (PK)ParkinsonsJ. Exp. Med.2012http://jem.rupress.org/content/209/4/837.short837-854209422473957Parkinson's disease (PD) is a progressive, chronic disease characterized by dyskinesia, rigidity, instability, and tremors. The disease is defined by the presence of Lewy bodies, which primarily consist of aggregated α-synuclein protein, and is accompanied by the loss of monoaminergic neurons. Current therapeutic strategies only give symptomatic relief of motor impairment and do not address the underlying neurodegeneration. Hence, we have identified Cu(II)(atsm) as a potential therapeutic for PD. Drug administration to four different animal models of PD resulted in improved motor and cognition function, rescued nigral cell loss, and improved dopamine metabolism. In vitro, this compound is able to inhibit the effects of peroxynitrite-driven toxicity, including the formation of nitrated α-synuclein oligomers. Our results show that Cu(II)(atsm) is effective in reversing parkinsonian defects in animal models and has the potential to be a successful treatment of PD.10.892Animals had access to water ad libitum throughout the pre- and postdose sampling period. Arterial blood samples were collected over the 24 h postdose period by a Culex Automated Blood Sampler (Bioanalytical Systems Inc.) via an indwelling carotid arterial cannula inserted under surgical anesthesia on the day before dosing. Serial blood collection was performed for individual rats up to 24 h after dose administration directly into heparinized borosilicate vials maintained at 4°C; blood samples were centrifuged and aliquots of plasma were stored frozen at −20°C until analysis by liquid chromatography-mass spectrometry (LC-MS)2012-04-09 00:00:0010.1084/jem.20112285
The influence of sertraline on the pharmacokinetics of carvedilol. A preclinical study on ratsAbrudan, MB;Muntean, DM;Neag, MA;Vlase, L;PK/PDDrug-drug InteractionFARMACIA2017https://pdfs.semanticscholar.org/d3b5/69049c147dbd82316db396626b9b1f652008.pdf557-562Prior to being connected to BASi Culex ABC®, each rat had his left femoral vein cannulated in order to allow precise Martignoni M, Groothuis GMM, de Kante R. Species differences between mouse, rat, dog, monkey and human CYP-mediated drug metabolism, inhibition and2017-01-01 00:00:00
The Pharmacokinetic Interaction Study between Carvedilol and Bupropion in RatsAbrudan, MB;Muntean, DM;Gheldiu, AM;Neag, MA;Vlase, L;Pharmacokinetics (PK)Drug-drug InteractionPharmacology2017https://www.karger.com/Article/Abstract/453619139-143993-428052289The effects of multiple-dose bupropion on the pharmacokinetics of single-dose carvedilol were investigated in order to evaluate this possible drug-drug interaction. A preclinical study was conducted among white male Wistar rats. Each rat was cannulated on the femoral vein prior to being connected to BASi Culex ABC®. During the reference period, each rat received an intravenous and an oral dose of 3.57 mg/kg body weight (b.w.) carvedilol, at 2 days distance. After 5 days of pretreatment with 21.42 mg/kg b.w. bupropion (by oral route, twice a day - given in order to reach the steady state), during the sixth day, 3.57 mg/kg b.w. carvedilol and 21.42 mg/kg b.w. bupropion were orally co-administrated (test period). After each administration of carvedilol, several samples of 200 µL blood were collected. The pharmacokinetic parameters of carvedilol were analyzed by the noncompartmental method. The 5 days pretreatment with bupropion increased the exposure to carvedilol in rats by 180%, considering the modifications observed in the area under the curve of carvedilol. Carvedilol was shown to have higher plasma concentrations, delay in maximum concentration, and a prolonged half-life, after being pretreated with bupropion. The administration of multiple-dose bupropion influences the pharmacokinetics of carvedilol (single oral dose) in rats. © 2017 S. Karger AG, Basel.1.615Methods: A preclinical study was conducted among white male Wistar rats. Each rat was cannulated on the femoral vein prior to being connected to BASi Culex ABC® Each rat was cannulated on the left femoral vein, prior to being connected to BASi Culex ABC®2017-01-05 00:00:0010.1159/000453619
The use of mass spectrometry to analyze dried blood spotsWagner, M;Tonoli, D;Varesio, E;Hopfgartner, G;DBSData ReviewMass Spectrom Rev2016https://onlinelibrary.wiley.com/doi/abs/10.1002/mas.21441361-43835325252132Dried blood spots (DBS) typically consist in the deposition of small volumes of capillary blood onto dedicated paper cards. Comparatively to whole blood or plasma samples, their benefits rely in the fact that sample collection is easier and that logistic aspects related to sample storage and shipment can be relatively limited, respectively, without the need of a refrigerator or dry ice. Originally, this approach has been developed in the sixties to support the analysis of phenylalanine for the detection of phenylketonuria in newborns using bacterial inhibition test. In the nineties tandem mass spectrometry was established as the detection technique for phenylalanine and tyrosine. DBS became rapidly recognized for their clinical value: they were widely implemented in pediatric settings with mass spectrometric detection, and were closely associated to the debut of newborn screening (NBS) programs, as a part of public health policies. Since then, sample collection on paper cards has been explored with various analytical techniques in other areas more or less successfully regarding large-scale applications. Moreover, in the last 5 years a regain of interest for DBS was observed and originated from the bioanalytical community to support drug development (e.g., PK studies) or therapeutic drug monitoring mainly. Those recent applications were essentially driven by improved sensitivity of triple quadrupole mass spectrometers. This review presents an overall view of all instrumental and methodological developments for DBS analysis with mass spectrometric detection, with and without separation techniques. A general introduction to DBS will describe their advantages and historical aspects of their emergence. A second section will focus on blood collection, with a strong emphasis on specific parameters that can impact quantitative analysis, including chromatographic effects, hematocrit effects, blood effects, and analyte stability. A third part of the review is dedicated to sample preparation and will consider off-line and on-line extractions; in particular, instrumental designs that have been developed so far for DBS extraction will be detailed. Flow injection analysis and applications will be discussed in section IV. The application of surface analysis mass spectrometry (DESI, paper spray, DART, APTDCI, MALDI, LDTD-APCI, and ICP) to DBS is described in section V, while applications based on separation techniques (e.g., liquid or gas chromatography) are presented in section VI. To conclude this review, the current status of DBS analysis is summarized, and future perspectives are provided. © 2014 Wiley Periodicals, Inc.9.068Consequently, blood spotting has to be performed manually in a second step. Wong et al. (2011) demonstrated the use of such an instrument (Culex, BASi) for serial sampling of a mouse and determination of a 24‐hr PK profile of a drug candidate2016-09-22 00:00:0010.1002/mas.21441
Theme VIII–Refinement and WelfareHoness, P;Baboo, E;Griffiths, MA;Safety PharmacologyMethod DevelopmentTraining2014http://www.altex.ch/resources/249273_WC9_Theme_8.pdf1-25Paul, MN, USA; 2toxicology, BASi, Mount Vernon, IN, USA dsarazan@datasci.com this study established the feasibility of collecting simultaneous blood samples by the auto- mated Culex-l system while recording Session VIII-2a: Best practice welfare approaches - Mouse2014-01-01 00:00:00
Thinking about dried blood spots for pharmacokinetic assays and therapeutic drug monitoringKissinger, PT;DBSData ReviewBioanalysis2011https://www.future-science.com/doi/abs/10.4155/BIO.11.2352263-2266320220111732.321declares the possibility of conflicts in his role as founder and Chairman Emeritus of BASi; former CEO Mass Spec.22(9),1501-1507 (2011).Crossref, Medline, CAS, Google Scholar; 101 Bioanalytical Systems. www.basinc.com/products/culex/Google Scholar; 102 Eli Lilly & Co2011-10-01 00:00:0010.4155/bio.11.235
Time-series responses of swine plasma metabolites to ingestion of diets containing myo-inositol or phytaseCowieson, AJ;Roos, FF;Ruckebusch, JP;Wilson, JW;Guggenbuhl, P;Lu, H;Ajuwon, KM;Adeola, O;Pharmacokinetics (PK)Nutritional SupplementsBr. J. Nutr.2017https://www.cambridge.org/core/journals/british-journal-of-nutrition/article/timeseries-responses-of-swine-plasma-metabolites-to-ingestion-of-diets-containing-myoinositol-or-phytase/6F4A8A7A962061E10BF87071A3EBED2A897-9051181129173209The effect of the ingestion of diets containing either myo-inositol or exogenous phytase on plasma metabolites was examined using 29 kg barrows. The diets were: control (maize, soya, rapeseed, rice bran), control plus 2 g/kg myo-inositol, control plus 1000 phytase units (FYT)/kg or 3000 FYT/kg exogenous phytase. Pigs were housed in a PigTurn device and blood was collected, from jugular catheters, via an automated system at -30, (30 min before feeding), 0, 15, 30, 45, 60, 90, 120, 150, 180, 240, 300 and 360 min post-feeding. The addition of 2 g/kg myo-inositol to the basal diet resulted in an increase in plasma myo-inositol concentration that was evident 45-60 min after diet introduction and persisted to 360 min post-feeding. Similarly, supplementation of the basal diet with either 1000 or 3000 FYT/kg exogenous phytase resulted in an increase in plasma myo-inositol concentration that was still rising 360 min post-feeding. Plasma P concentration was increased over time by the addition of 1000 and 3000 FYT/kg phytase, but not by the addition of myo-inositol. Other plasma metabolites examined were not affected by dietary treatment. It can be concluded that oral delivery of myo-inositol results in rapid increase in plasma myo-inositol concentrations that peak approximately 45-60 min after feeding. Use of supplemental phytase achieves similar increases in myo-inositol concentration in plasma but the appearance is more gradual. Furthermore, supplementation of pig diets with exogenous phytase results in rapid appearance of P in plasma that may be sustained over time relative to diets with no added phytase.3.319All surgeries for each pig took approximately 45min. Blood collection 1 and 2). The catheter was attached to a 2-m catheter extension fixed to the automatic sampling system (Culex-L; BASi), which was programmed to collect blood at the collection intervals: −30, (30 min before2017-12-01 00:00:0010.1017/S0007114517003026
Topical application of a novel oxycodone gel formulation (tocopheryl phosphate mixture) in a rat model of peripheral inflammatory pain produces localized pain relief without significant systemic exposureSmith, MT;Wyse, BD;Edwards, SR;El-Tamimy, M;Gaetano, G;Gavin, P;Pharmacokinetics (PK)Pain ManagementJ Pharm Sci2015https://www.sciencedirect.com/science/article/pii/S00223549153006542388-2396104725995048This study was designed to assess the analgesic efficacy and systemic exposure of oxycodone administered topically in a novel tocopheryl phosphate mixture (TPM) gel formulation, to the inflamed hindpaws in a rat model of inflammatory pain. Unilateral hindpaw inflammation was induced in male Sprague-Dawley rats by intraplantar (i.pl.) injection of Freund's complete adjuvant (FCA). Mechanical hyperalgesia and hindpaw inflammation were assessed by measuring paw pressure thresholds and hindpaw volume, respectively, just prior to i.pl. FCA and again 5-6 days later. The analgesic effects of oxycodone administered topically (1 mg in TPM gel) or by i.pl. injection (50 μg), were assessed. Systemic oxycodone exposure was assessed over an 8-h postdosing interval following topical application. Skin permeation of oxycodone from the gel formulation was assessed in vitro using Franz diffusion cells. Oxycodone administered topically or by i.pl. injection produced significant (p < 0.05) analgesia in the inflamed hindpaws. Systemic oxycodone exposure was insignificant after topical dosing. The in vitro cumulative skin permeation of oxycodone was linearly related to the amount applied. Topical TPM/oxycodone gel formulations have the potential to alleviate moderate to severe inflammatory pain conditions with minimal systemic exposure, thereby avoiding central nervous system (CNS)-mediated adverse effects associated with oral administration of opioid analgesics. © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association.3.197Each rat received 2-3 randomly assigned oxycodone doses (n = 2-6 per dose) or vehicle (n = 5). PPT values for each hindpaw were An 1-cm, incision was made above the dorsal aspect of the scapulae to allow insertion of a BASi Culex® (West Lafayetta, Indiana, USA) portal 2015-07-01 00:00:0010.1002/jps.24502
Toxicokinetics and hydrolysis of artelinate and artesunate in malaria-infected ratsLi, Q;Xie, LH;Si, Y;Wong, E;Upadhyay, R;Yanez, D;Weina, PJ;PK/PDAntimalarialInt. J. Toxicol.2005https://journals.sagepub.com/doi/abs/10.1080/10915810591007201241-25024416126618Comparative toxicokinetic (TK) and hydrolysis studies of intravenously administered two new antimalarial agents, artelinate (AL) and artesunate (AS), were performed in malaria-infected rats using three daily equimolar doses (96 micromoles/kg). The TK evaluation was related to select one drug for severe malaria treatment in U.S. Army. Drug concentration of AS with daily dose of 36.7 mg/kg was one-third less on day 3 than on day 1, which resembled its active metabolite, dihydroartemisinin (DHA), suggesting an autoinduction of hepatic drug-metabolizing enzymes for AS. The results were similar to other artemisinin drugs, but not for AL. TK parameters of AL were very comparable from day 1 to day 3 at same AS molecular dose at 40.6 mg/kg. AS is the prodrug of DHA with the DHA/AS ratio of 5.26 compared to the ratio of 0.01 for DHA/AL. Other TK parameters revealed that the total AUC1-3 days (84.4 microg.h ml-1) of AL was fivefold higher than that of AS (15.7 microg.h ml-1 of AS plus DHA). The elimination half-life of AL (7.1 h) was much longer than that of AS (0.36 h) or DHA (0.72 h). The remarkable alteration of the TK shape of AL may be caused by poor conversion rates to DHA and an enterohepatic circulation, which is confirmed by the present TK and tissue distribution studies. Compared to AS, higher drug exposure levels and longer exposure time of AL in the rat blood may be the cause of its increased toxicity.1.223The Culex ABS is a blood sampling and metabolism monitoring machine, which is controlled by its own internal computer. We were successful in evaluating TK for long-term periods by using this system. After jugular vein cannulation, study animals were individually placed in the system, which allowed for a micropump to infuse heparinized saline (10 units heparin ml−1 saline) at a rate of 1 to 2 µl/min to prevent coagulation within the catheter. Culex obtained precise blood volumes from the study animals and maintained total collected volume at less than 6% per day and 14% during the entire study period. Blood samples were maintained at 3◦C using Culex’s temperature-controlled fractional collector. Plasma was separated from the blood samples and processed for drug extraction. The detailed sampling process was described previously (Reisen, Lothrop, and Meyer 1997; Tian et al. 2002)2005-08-30 00:00:0010.1080/10915810591007201
Tracking deposition of a 14C-radiolabeled kudzu hairy root-derived isoflavone-rich fraction into boneMun, JG;Grannan, MD;Lachcik, PJ;Rogers, RB;Yousef, GG;Grace, MH;Janle, EM;Wu, QL;Simon, JE;Weaver, CM;Lila, MA;PK and MicrodialysisNutritional SupplementsExp. Biol. Med. (Maywood)2010https://journals.sagepub.com/doi/abs/10.1258/ebm.2010.0101341224-12352351020851832Hairy roots were induced in four genotypes from three kudzu species (Pueraria montana var. lobata, P. lobata and P. phaseoloides) in vitro using Agrobacterium rhizogenes to stimulate rapid secondary metabolite synthesis. Hairy roots from P. montana var. lobata (United States Department of Agriculture no. PI 434246) yielded the highest puerarin and total isoflavone content and the greatest new biomass per growth cycle among the genotypes evaluated. Hairy roots from this genotype were selected for radiolabeling using (14)C-sucrose as a carbon source. Isoflavones from radiolabeled kudzu hairy root cultures were extracted with 80% methanol, partitioned by solvent extraction, and then subfractionated by Sephadex LH-20 gel filtration. Radiolabeled isoflavones were isolated in a highly enriched fraction, which contained predominantly puerarin, daidzin and malonyl-daidzin and had an average radioactivity of 8.614 MBq/g (232.8 μCi/g) dry fraction. The (14)C-radiolabeled, isoflavone-rich fraction was orally administered at a dose of 60 mg/kg body weight to male Sprague-Dawley rats implanted with a jugular catheter, a subcutaneous ultrafiltrate probe and a brain microdialysate probe. Serum, interstitial fluid, brain microdialysate, urine and feces were collected using a Culex(®) Automated Blood Collection System for 24 h. At the end of this period, rats were sacrificed and major tissues were collected. Analysis by a scintillation counter confirmed that a bolus dose of (14)C-radiolabeled, isoflavone-rich kudzu fraction reached bone tissues, which accumulated 0.011%, 0.09% and 0.003% of the administered dose in femur, tibia and vertebrae, respectively. Femurs extracted with 80% methanol were analyzed by high-performance liquid chromatography with electrospray ionization-mass spectrometry and were found to contain trace quantities of puerarin, daidzein and puerarin glucuronide. This study demonstrates that kudzu isoflavones and metabolites are capable of reaching bone tissues, where they may contribute to the prevention of osteoporosis and the promotion of bone health.Hairy roots were induced in four genotypes from three kudzu species (Pueraria montana var. lobata, P. lobata and P. phaseoloides) in vitro using Agrobacterium rhizogenes to stimulate rapid secondary metabolite synthesis. Hairy roots from P. montana var. lobata (United States Department of Agriculture no. PI 434246) yielded the highest puerarin and total isoflavone content and the greatest new biomass per growth cycle among the genotypes evaluated. Hairy roots from this genotype were selected for radiolabeling using (14)C-sucrose as a carbon source. Isoflavones from radiolabeled kudzu hairy root cultures were extracted with 80% methanol, partitioned by solvent extraction, and then subfractionated by Sephadex LH-20 gel filtration. Radiolabeled isoflavones were isolated in a highly enriched fraction, which contained predominantly puerarin, daidzin and malonyl-daidzin and had an average radioactivity of 8.614 MBq/g (232.8 μCi/g) dry fraction. The (14)C-radiolabeled, isoflavone-rich fraction was orally administered at a dose of 60 mg/kg body weight to male Sprague-Dawley rats implanted with a jugular catheter, a subcutaneous ultrafiltrate probe and a brain microdialysate probe. Serum, interstitial fluid, brain microdialysate, urine and feces were collected using a Culex(®) Automated Blood Collection System for 24 h. At the end of this period, rats were sacrificed and major tissues were collected. Analysis by a scintillation counter confirmed that a bolus dose of (14)C-radiolabeled, isoflavone-rich kudzu fraction reached bone tissues, which accumulated 0.011%, 0.09% and 0.003% of the administered dose in femur, tibia and vertebrae, respectively. Femurs extracted with 80% methanol were analyzed by high-performance liquid chromatography with electrospray ionization-mass spectrometry and were found to contain trace quantities of puerarin, daidzein and puerarin glucuronide. This study demonstrates that kudzu isoflavones and metabolites are capable of reaching bone tissues, where they may contribute to the prevention of osteoporosis and the promotion of bone health2010-10-01 00:00:0010.1258/ebm.2010.010134
Use of trifluoroacetic acid to quantify small, polar compounds in rat plasma during discovery-phase pharmacokinetic evaluationBock, MJ;Neilson, KL;Dudley, A;Pharmacokinetics (PK)Method DevelopmentJ. Chromatogr. B Analyt. Technol. Biomed. Life Sci.2007https://www.sciencedirect.com/science/article/pii/S1570023207003856165-1708561-217581800Although it is accepted that trifluoroacetic acid (TFA) can cause suppression of an analyte during LC/MS analysis, this paper presents a relatively sensitive gradient method that uses a TFA mobile phase for the improved quantification of small, polar drug-like compounds. The described method was developed in a discovery drug metabolism and pharmacokinetics (DMPK) laboratory for the screening measurement of compound concentrations to calculate PK parameters and CNS exposure of compounds from a chemical series that had poor chromatography under generic methods using formic acid mobile phase. The samples were collected by a Culex automated sampling unit, and the plasma proteins were precipitated by a Tecan robot in 96-well plates. After centrifugation, the supernatant was removed, dried down using a SPE-Dry unit, and the samples were reconstituted in aqueous buffer on the robot. The samples were analyzed on an Agilent LC/MSD using a 5-min gradient on a 5 cm phenyl column. No additional steps, such as the "TFA-fix", were necessary. Although sample batches were analyzed over 6h, no drift or degradation of signal was observed. The improved chromatography resulted in a method that was selective, rugged, and had a dynamic range from 5 to 20,000 nM, which was sufficient to quantitate low volume, serial plasma samples collected out to 8 h postdose.2.813Because DMPK pharmacokinetic samples are usually collected by a Culex ABS system (BAS, West Lafayette This method was used in rat pharmacokinetic screening studies for the quantification of samples out to 6 h following a 10 μmol/kg IV dose and 10 h following a 30 2007-09-01 00:00:0010.1016/j.jchromb.2007.05.024
Using miniature MS system with automatic blood sampler for preclinical pharmacokinetics studyPu, F;Zhang, W;Bateman, KP;Liu, Y;Helmy, R;Ouyang, Z;Pharmacokinetics (PK)Method DevelopmentBioanalysis2017https://www.future-science.com/doi/abs/10.4155/bio-2017-01601633-164192129095035Preclinical pharmacokinetic studies are an essential part of modern drug development. In this work, we explored a new solution with onsite analysis using a miniature MS system, which can significantly improve the efficiency of the preclinical study. Materials & methods: A miniature mass spectrometer was used with an automatic blood sampler for onsite quantitation of drug compounds in whole blood samples. Slug-flow microextraction was used to replace the in-lab sample preparation. Animal studies were carried out using two drug compounds, using the auto sampler to take blood samples at preprogrammed time points. The miniature MS system was used to obtain drug concentrations, which were subsequently used to calculate the pharmacokinetic parameters.2.321which was manufactured by BASi, Inc. Culex ABS was capable of sampling rodent blood automatically at programmed time points. Animal studies. Sprague Dawley male rats were purchased from Envigo (Huntington, UK). A catheter was implanted into each rat's artery2017-11-01 00:00:0010.4155/bio-2017-0160
Utilization of DBS within drug discovery: development of a serial microsampling pharmacokinetic study in miceClark, GT;Haynes, JJ;Bayliss, MA;Burrows, L;DBSMethod DevelopmentBioanalysis2010https://www.future-science.com/doi/abs/10.4155/bio.10.911477-14882821083348Dried blood spots (DBS) are rapidly gaining a foothold in the pharmaceutical industry. However, applications in exploratory drug discovery are limited mainly owing to method development time. The development of a generic DBS assay is presented with its application in serial microsampling from mice. A generic 'fit-for-purpose' assay was developed on FTA(®) Elute, which allowed 90% of compounds tested to reach sensitivity levels of 1 ng/ml. A ten-time point serial mouse pharmacokinetic study was conducted using 20-µl microsamples and DBS. Application of generic 'fit-for-purpose' approach did not compromise data delivery or quality. Serial microsampling and DBS in exploratory mouse pharmacokinetic has been shown to provide superior data quality when compared with traditional plasma-based composite studies.2.321The authors see the development of this technology over the next 5 years to be in two areas: ▪ The continued refinement of sample collection, in particular automated blood collection systems such as the Accusampler™ (DiLab) or Culex (BASi) system for microsampling; ; In our laboratories we traditionally run exploratory rat PK studies from n = 2 animals with The technique was qualified by running composite and serial sampling mouse studies in automated blood collection systems such as the Accusampler™ (DiLab) or Culex (BASi) system for2010-08-01 00:00:0010.4155/bio.10.91
Utilization of Stable Isotope Labeling to Facilitate the Identification of Polar Metabolites of KAF156, an Antimalarial AgentHuskey, SE;Forseth, RR;Li, H;Jian, Z;Catoire, A;Zhang, J;Ray, T;He, H;Flarakos, J;Mangold, JB;Pharmacokinetics (PK)AntimalarialDrug Metab. Dispos.2016http://dmd.aspetjournals.org/content/44/10/1697.short1697-1708441027486238Identification of polar metabolites of drug candidates during development is often challenging. Several prominent polar metabolites of 2-amino-1-(2-(4-fluorophenyl)-3-((4-fluorophenyl)amino)-8,8-dimethyl-5,6-dihydroimidazo[1,2-a]pyrazin-7(8H)-yl)ethanone ([(14)C]KAF156), an antimalarial agent, were detected in rat urine from an absorption, distribution, metabolism, and excretion study but could not be characterized by liquid chromatography-tandem mass spectrometry (LC-MS/MS) because of low ionization efficiency. In such instances, a strategy often chosen by investigators is to use a radiolabeled compound with high specific activity, having an isotopic mass ratio (i.e., [(12)C]/[(14)C]) and mass difference that serve as the basis for a mass filter using accurate mass spectrometry. Unfortunately, [(14)C]KAF156-1 was uniformly labeled (n = 1-6) with the mass ratio of ∼0.1. This ratio was insufficient to be useful as a mass filter despite the high specific activity (120 μCi/mg). At this stage in development, stable isotope labeled [(13)C6]KAF156-1 was available as the internal standard for the quantification of KAF156. We were thus able to design an oral dose as a mixture of [(14)C]KAF156-1 (specific activity 3.65 μCi/mg) and [(13)C6]KAF156-1 with a mass ratio of [(12)C]/[(13)C6] as 0.9 and the mass difference as 6.0202. By using this mass filter strategy, four polar metabolites were successfully identified in rat urine. Subsequently, using a similar dual labeling approach, [(14)C]KAF156-2 and [(13)C2]KAF156-2 were synthesized to allow the detection of any putative polar metabolites that may have lost labeling during biotransformations using the previous [(14)C]KAF156-1. Three polar metabolites were thereby identified and M43, a less polar metabolite, was proposed as the key intermediate metabolite leading to the formation of a total of seven polar metabolites. Overall this dual labeling approach proved practical and valuable for the identification of polar metabolites by LC-MS/MS. Copyright © 2016 by The American Society for Pharmacology and Experimental Therapeutics.3.354Male Wistar Hannover rats (∼250–320 g, ∼8 week, n = 24) were from Harlan Laboratories (Somerville, NJ). Catheters were surgically implanted into the carotid artery and/or jugular vein of rats by the vendor (only one catheter was implanted into carotid artery for blood collection from rats receiving oral dose). All rats were housed individually in metabolism cages (Culex Autosampler, BAS, Indianapolis, IN) in a temperature and humidity controlled room with free access to food and water (food was withheld until 4 hours postdose)2016-10-01 00:00:0010.1124/dmd.116.072108
Your bioanalytical data are only as good as your samplesKissinger, CB;Kissinger, PT;Pharmacokinetics (PK)Data ReviewBioanalysis2012https://www.future-science.com/doi/abs/10.4155/bio.12.1251411-1415412227930252.321As a result, automatically collecting serial samples from an awake mouse is now feasible, and in some laboratories very routine www.davita.comGoogle Scholar; 106 BASi. www.basinc.com/products/culexGoogle Scholar; 107 Prosolia2012-06-01 00:00:0010.4155/bio.12.125