Bhagwat Prasad

Assistant Professor

Department of Pharmaceutics, Pharmaceutics Faculty, UWPKDAP, UWRAPT

Telephone: (206) 221-2295

Email: bhagwat@uw.edu

Office Location: H268, Health Science Building

Website: Prasad lab

Education

  • Ph.D. and Master of Science in Pharmaceutical Analysis, National Institute of Pharmaceutical Education and Research (NIPER), SAS Nagar, India
  • Bachelor in Pharmaceutical Sciences, HNB Garhwal University, India

Research Interests

The overarching objective of our research is to investigate mechanisms of age-dependent variability in xeno- and endo-biotic disposition. Our current research projects involve the use of state-of-the-art LC-MS/MS quantitative proteomics and metabolomics to elucidate inter-individual differences in the abundances and activities of non-cytochrome P450 (non-CYP) drug metabolizing enzymes and transporters (DMET) in human tissues. We then integrate these data into physiologically-based pharmacokinetic (PBPK) models to predict variability in drug disposition. Such approaches are particularly important to predict drug disposition in special populations such as pediatrics where clinical studies are not often feasible. Currently, we have four specific projects undergoing in our laboratory.

  1. Development and validation of DMET quantitative proteomics methods: We apply robust LC-MS/MS proteomics approach to quantify DMET proteins in human and animal tissues, subcellular fractions and cells. We have developed a repository of LC-MS/MS methods for quantification of >280 human DMET proteins. We are also developing novel non-invasive methods for predicting drug disposition, e.g., by quantifying drug transporters in exosomes. These methods are being utilized to quantify effect of age, genotype and disease condition on expression of DMET proteins.
  2. PBPK prediction of ontogeny mediated hepatic and renal drug disposition: Our NICHD funded research utilizes in-house generated protein quantification data of hepatic DMET proteins to build PBPK models for prediction of age-dependent hepatic drug disposition. Such approaches are critical in determining first-in-children dose. We are also investigating age-dependent pharmacokinetics and pharmacodynamics of diuretics.
  3. Effect of UGT2B17 variability on drug and sex-steroid disposition: We are investigating the effect of age, gender and genetic polymorphism related variability of UGT2B17 on metabolism of typical UGT substrates including sex-steroids such as testosterone, dihydrotestosterone and estradiol.
  4. Integrated proteomics and metabolomics analysis of the steroidogenic pathway: We are also investigating age-dependent changes in steroidogenic pathway with respect to the role of hepatic and intestinal DMET proteins during human development. The quantitative proteomics and metabolomics assays developed in our laboratory are used to better elucidate mechanisms of fluctuations in levels of steroidal hormones.

 

Biography

I am a pharmacist by education with major research training in the field of pharmaceutical analysis. My recent research is focused on application of liquid chromatography-mass spectrometry (LC-MS) and quantitative proteomics to understand interindividual variability in drug disposition. Before joining as an assistant professor at UW Pharmaceutics, I worked with Prof. Jash Unadkat as a postdoc and a lead scientist of UW Research Affiliate Program on Transporters (UWRAPT). I obtained my MS in 2006 and Ph.D. in 2010 in Pharmaceutical Sciences from NIPER, Mohali, India. In my role as a PI or co-Inv. in various NIH and industry funded grants, I have been establishing and applying quantitative proteomic assays to quantify population variability in DMET proteins

Selected Publications

PubMed

  1. Bhatt DK, Gaedigk A, Pearce R, Leeder S, Prasad B*, Age-dependent Protein Abundance of Cytosolic Alcohol and Aldehyde Dehydrogenases in Human Liver, Drug Metab and Dispos, 2017.
  2. Xu M, Bhatt DK, Yeung CK, Claw KG, Singh A, Gaedigk A, Pearce R, Gaedigk R, Nickerson D, Schuetz E, Rettie AE, Leeder SJ, Thummel KE, Prasad B*, Genetic and Non-genetic Factors Associated with Protein Abundance of Flavin-containing Monooxygenase 3 in Human Liver, Accepted for publication in J Pharmacol Exp Ther, 2017.
  3. Bhatt DK and Prasad B*, Critical issues and optimized practices in quantification of protein abundance level to determine inter-individual variability in DMET proteins by LC-MS/MS proteomics, Accepted for publication in Clin Pharmacol Ther, 2017 (Review article).
  4. Boberg M, Vrana M, Mehrotra A, Pearce R, Gaedigk A, Leeder JS, Prasad B*. Age dependent absolute abundance of hepatic microsomal carboxylesterases (CES1 and CES2) by LC-MS/MS proteomics: Application in prediction of in vivo pharmacokinetics of oseltamivir, Drug Metabolism and Disposition, 2016.
  5. Liao MZ, Gao C, Shireman LM, Phillips B, Risler LJ, Neradugomma NK, Choudhari P, Prasad B, Shen DD, Mao Q. P-gp/ABCB1 exerts differential impacts on brain and fetal exposure to norbuprenorphine. Pharmacol Res. 2017.
  6. Prasad B, Johnson K, Billington S, Lee CA, Chung GW, Brown CDA, Kelly E, Himmelfarb J, Unadkat JD. Abundance of drug transporters in the human kidney cortex as quantified by LC-MS/MS proteomics Drug Metabolism and Disposition, 2016 (dmd. 116.072066).
  7. Vrana M, Goodling A, Afkarian M, Prasad B*, An optimized method for protein extraction from OCT-embedded human kidney tissue for protein quantification by LC-MS/MS proteomics” Drug Metab and Dispos, 2016:44, 1692-1696.
  8. Prasad B*., Gaedigk A., Vrana N., Gaedigk R., Leeder J.S., Salphati L., Chu X., Xiao G., Hop C.E.C.A., Evers R., Gan L., Unadkat J.D.,Ontogeny of Hepatic Drug Transporters as Quantified by LC-MS/MS Proteomics, Clin Pharmacol Therap [Epub ahead of print], 2016.
  9. Singh S, Prasad B, Savaliya A, Shah R, Gohil V, Kaur A. Strategies for characterizing sildenafil, vardenafil, tadalafil and their analogues in herbal dietary supplements, and detecting counterfeit products containing these drugs. Trends in Analytical Chemistry. 2009;28:13-28.
  10. Prasad B, Garg A, Takwani H, Singh S. Metabolite identification by liquid chromatography-mass spectrometry. Trends in Analytical Chemistry. 2011;30:360-87.
  11. Prasad B, Evers R, Gupta A, Hop CE, Salphati L, Shukla S, et al. Interindividual Variability in Hepatic Oatps and P-Glycoprotein (ABCB1) Protein Expression: Quantification by LC-MS/MS and Influence of Genotype, Age and Sex. Drug Metab Dispos. 2013 Oct 11.
  12. Prasad B, Lai Y, Lin Y, Unadkat JD. Interindividual variability in the hepatic expression of the human breast cancer resistance protein (BCRP/ABCG2): effect of age, sex, and genotype. J Pharm Sci. 2013 Mar;102(3):787-93.
  13. Deo AK, Prasad B, Balogh L, Lai Y, Unadkat JD. Interindividual variability in hepatic expression of the multidrug resistance-associated protein 2 (MRP2/ABCC2): quantification by liquid chromatography/tandem mass spectrometry. Drug Metab Dispos. 2012 May;40(5):852-5.
  14. Prasad B*, Unadkat JD. Optimized Approaches for Quantification of Drug Transporters in Tissues and Cells by MRM Proteomics. AAPS J. 2014 Jul;16(4):634-48.
  15. Prasad B, Unadkat JD, The concept of fraction of drug transported (ft) with special emphasis on BBB efflux of CNS and antiretroviral drugs, 2015, Clin Pharmacol Therap (DOI: 10.1002/cpt.72).
  16. Kumar V, Prasad B, Patilea G, Gupta A, Salphati L, Evers R, Hop CECA, Unadkat JD, Quantitative transporter proteomics by LC-MS/MS: addressing methodological issues of plasma membrane isolation and expression-activity relationship, 2015, Drug Metab Dispos, dmd. 114.061614.
  17. Arnold SL, Kent T, Hogarth CA, Schlatt S, Prasad B, Haenisch M, Walsh, T, Muller CH, Griswold MD, Amory JK, Isoherranen N, Importance of ALDH1A enzymes in determining human testicular retinoic acid concentrations, Journal of lipid research,. M054718
  18. Wang L, Prasad B, Salphati L, Chu X, Gupta A, Hop CECA, Evers R, Unadkat JD, Interspecies Variability in Expression of Hepatobiliary Transporters across Human, Dog, Monkey, and Rat as Determined by Quantitative Proteomics, Drug Metab Dispos, 2015 43 (3), 367-374
  19. Shuster DL, Risler LJ, Prasad B, Calamia JC, Voellinger JL, Kelly EJ, et al. Identification of CYP3A7 for glyburide metabolism in human fetal livers. Biochem Pharmacol. 2014 92 (4), 690-700.
  20. Stieger B, Unadkat JD, Prasad B, Langer O, Gali H. Role of (drug) transporters in imaging in health and disease. Drug Metab Dispos. 2014 Dec;42(12):2007-15.
  21. Salphati L, Chu X, Chen L, Prasad B, Dallas S, Evers R, et al. Evaluation of Organic Anion Transporting Polypeptide 1B1 and 1B3 Humanized Mice as a Translational Model to Study the Pharmacokinetics of Statins. Drug Metab Dispos. 2014 Aug;42(8):1301-13.
  22. Prasad B, Unadkat JD. Comparison of Heavy Labeled (SIL) Peptide versus SILAC Protein Internal Standards for LC-MS/MS Quantification of Hepatic Drug Transporters. Int J Proteomics. 2014;2014:451510.
  23. He J, Yu Y, Prasad B, Link J, Miyaoka RS, Chen X, et al. PET Imaging of Oatp-Mediated Hepatobiliary Transport of [C] Rosuvastatin in the Rat. Mol Pharm. 2014 Jul 8.
  24. He J, Yu Y, Prasad B, Chen X, Unadkat JD. Mechanism of an unusual, but clinically significant, digoxin-bupropion drug interaction. Biopharm Drug Dispos. 2014 Jul;35(5):253-63.
  25. Drozdzik M, Groer C, Penski J, Lapczuk J, Ostrowski M, Lai Y, et al. Protein Abundance of Clinically Relevant Multidrug Transporters along the Entire Length of the Human Intestine. Mol Pharm. 2014 Oct 6;11(10):3547-55.
  26. Prasad B. Targeted MRM Proteomics is a Better Protein Quantification Method Over Western-Blotting. J Anal Bioanal Tech. 2013;5:e117.
  27. Lee N, Hebert MF, Prasad B, Easterling TR, Kelly EJ, Unadkat JD, et al. Effect of Gestational Age on mRNA and Protein Expression of Polyspecific Organic Cation Transporters during Pregnancy. Drug Metab Dispos. 2013 Oct 7.
  28. Edson KZ, Prasad B, Unadkat JD, Suhara Y, Okano T, Guengerich FP, et al. Cytochrome P450-Dependent Catabolism of Vitamin K: omega-Hydroxylation Catalyzed by Human CYP4F2 and CYP4F11. Biochemistry. 2013 Oct 18.
  29. Prasad B, Singh S. In vitro reaction phenotyping studies on rifamycins to explain the auto-induction of rifabutin metabolism. Int J Tuberc Lung Dis. 2012 Feb;16(2):232-4.
  30. Prasad B, Singh S. Identification of rat urinary metabolites of rifabutin using LC-MSn and LC-HR-MS. Eur J Pharm Sci. 2010 Sep 11;41(1):173-88.
  31. Prasad B, Singh S. In vitro and in vivo investigation of metabolic fate of rifampicin using an optimized sample preparation approach and modern tools of liquid chromatography-mass spectrometry. J Pharm Biomed Anal. 2009 Oct 15;50(3):475-90.
  32. Prasad B, Singh S. LC-MS/TOF and UHPLC-MS/MS study of in vivo fate of rifamycin isonicotinyl hydrazone formed on oral co-administration of rifampicin and isoniazid. J Pharm Biomed Anal. 2009 Jul 8;52(3):377-83.
  33. Mayatra SJ, Prasad B, Jain M, Jain R, Singh S. Development and validation of a sensitive and selective UHPLC-MS/MS method for quantitation of an investigational anti-malarial compound, 2-tert-butylprimaquine (NP-96) in rat plasma, and its application in a preclinical pharmacokinetic study. J Pharm Biomed Anal. 2009 Jul 8;52(3):410-5.
  34. Garg A, Prasad B. Emerging role of liquid chromatography mass spectrometric techniques in studying drug metabolism induced toxicity. Current Research & Information on Pharmaceutical Sciences (CRIPS). 2009;11:1-5.
  35. Prasad B, Kumar V, Bhutani H, Singh S. Screening of bulk drug samples and anti-tuberculosis products for the presence of therapeutically less active diasteriomeric (R,S) form of ethambutol dihydrochloride. Indian J Tuberc. 2008 Oct;55(4):192-8.
  36. Kumar V, Prasad B, Singh S. Polypill as a treatment strategy for cardiovascular diseases: Pharmaceutical issues in the development of a polypill for the treatment of cardiovascular diseases. Drug Discovery Today: Therapeutic Strategies. 2008;5:63-71.
  37. Prasad B, Bhutani H, Kumar V, Singh S. A new validated DSC method for monitoring of less active R,S isomer of ethambutol dihydrochloride in bulk drug samples and anti-tuberculosis formulations. Pharmacopeial Forum (USP) 2007;33:323-33.
  38. Prasad B, Bhutani H, Singh S. Study of the interaction between rifapentine and isoniazid under acid conditions. J Pharm Biomed Anal. 2006 Jun 16;41(4):1438-41.