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Influence of Ethanol on Darunavir Hepatic Clearance and Intracellular PK/PD in HIV-Infected Monocytes, and CYP3A4-Darunavir Interactions Using Inhibition and in Silico Binding Studies

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Abstract

Purpose

Although the prevalence of alcohol consumption is higher in HIV+ people than general public, limited information is available on how alcohol affects the metabolism and bioavailability of darunavir (DRV).

Methods

DRV was quantified by using LC-MS/MS method. All in vitro experiments were performed using human liver microsomes and HIV-infected monocytic cells. CYP3A4 and DRV/Ritonavir (RTV) docking was performed using GOLD suite 5.8.

Results

Ethanol (20 mM) significantly decreased apparent half-life and increased degradation rate constant of RTV-boosted DRV but not for DRV alone. Similarly, ethanol exposure increased hepatic intrinsic clearance for RTV-boosted DRV with no significant influence on DRV alone. Ethanol showed a limited influence on intracellular total DRV exposure in the presence of RTV without altering maximum concentration (Cmax) values in HIV-infected monocytic cells. Ethanol alone elevated HIV replication but this effect was nullified with the addition of DRV or DRV + RTV. Additionally, inhibitory potency of DRV was significantly reduced in the presence of ethanol. Our docking results projected that ethanol increases the average distance between DRV and CYP3A4 heme, and alter the orientation of DRV-CYP3A4 binding.

Conclusions

Collectively these findings suggest that DRV metabolism is primarily influenced by ethanol in the liver, but has minor effect in HIV-residing monocytes.

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Abbreviations

AUCtot :

Area under the concentration-time curve

Cmax :

The maximum concentration of a drug in the cells after dosing

CYP:

Cytochrome P450

DRV:

Darunavir

EVG:

Elvitegravir

IC50 :

The 50% inhibitory concentration

Km :

The Michaelis-Menten constant

LC-MS/MS:

Liquid chromatography-tandem mass spectrometry

RTV:

Ritonavir

Vmax :

The maximal velocity

References

  1. Pokorna J, Machala L, Rezacova P, Konvalinka J. Current and novel inhibitors of HIV protease. Viruses. 2009;1(3):1209–39.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Rittweger M, Arasteh K. Clinical pharmacokinetics of darunavir. Clin Pharmacokinet. 2007;46(9):739–56.

    Article  CAS  PubMed  Google Scholar 

  3. Mudra DR, Desino KE, Desai PV. In silico, in vitro and in situ models to assess interplay between CYP3A and P-gp. Curr Drug Metab. 2011;12(8):750–73.

    Article  CAS  PubMed  Google Scholar 

  4. Poizot-Martin I, Naqvi A, Obry-Roguet V, Valantin MA, Cuzin L, Billaud E, et al. Potential for drug-drug interactions between Antiretrovirals and HCV direct acting antivirals in a large cohort of HIV/HCV Coinfected patients. PLoS One. 2015;10(10):e0141164.

    Article  PubMed  PubMed Central  Google Scholar 

  5. Midde NM, Kumar S. Development of NanoART for HIV treatment: minding the cytochrome P450 (CYP) enzymes. J Pers Nanomed. 2015;1(1):24–32.

    PubMed  PubMed Central  Google Scholar 

  6. Chander G, Josephs J, Fleishman JA, Korthuis PT, Gaist P, Hellinger J, et al. Alcohol use among HIV-infected persons in care: results of a multi-site survey. HIV Med. 2008;9(4):196–202.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Kumar S, Jin M, Ande A, Sinha N, Silverstein PS, Kumar A. Alcohol consumption effect on antiretroviral therapy and HIV-1 pathogenesis: role of cytochrome P450 isozymes. Expert Opin Drug Metab Toxicol. 2012;8(11):1363–75.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Baum MK, Rafie C, Lai S, Sales S, Page JB, Campa A. Alcohol use accelerates HIV disease progression. AIDS Res Hum Retrovir. 2010;26(5):511–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Kumar S, Earla R, Jin M, Mitra AK, Kumar A. Effect of ethanol on spectral binding, inhibition, and activity of CYP3A4 with an antiretroviral drug nelfinavir. Biochem Biophys Res Commun. 2010;402(1):163–7.

    Article  CAS  PubMed  Google Scholar 

  10. Kumar S, Kumar A. Differential effects of ethanol on spectral binding and inhibition of cytochrome P450 3A4 with eight protease inhibitors antiretroviral drugs. Alcohol Clin Exp Res. 2011;35(12):2121–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Rao PS, Kumar S. Chronic effects of ethanol and/or Darunavir/ritonavir on U937 Monocytic cells: regulation of cytochrome P450 and antioxidant enzymes, oxidative stress, and cytotoxicity. Alcohol Clin Exp Res. 2016;40(1):73–82.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Midde NM, Rahman MA, Rathi C, Li J, Mieibohm B, Li W, et al. Effect of ethanol on the metabolic characteristics of HIV-1 integrase inhibitor Elvitegravir and Elvitegravir/Cobicistat with CYP3A: an analysis using a newly developed LC-MS/MS meth. PLoS One. 2016;11(2):e0149225.

  13. Rao P, Ande A, Sinha N, Kumar A, Kumar S. Effects of cigarette smoke condensate on oxidative stress, apoptotic cell death, and HIV replication in human Monocytic cells. PLoS One. 2016;11(5):e0155791.

    Article  PubMed  PubMed Central  Google Scholar 

  14. Earla R, Kumar S, Wang L, Bosinger S, Li J, Shah A, et al. Enhanced methamphetamine metabolism in rhesus macaque as compared with human: an analysis using a novel method of liquid chromatography with tandem mass spectrometry, kinetic study, and substrate docking. Drug Metab Dispos: Biolo Fate Chem. 2014;42(12):2097–108.

    Article  Google Scholar 

  15. Shelley JC, Cholleti A, Frye LL, Greenwood JR, Timlin MR, Uchimaya M. Epik: a software program for pK( a ) prediction and protonation state generation for drug-like molecules. J Comput Aided Mol Des. 2007;21(12):681–91.

    Article  CAS  PubMed  Google Scholar 

  16. Sekar V, Spinosa-Guzman S, De Paepe E, Stevens T, Tomaka F, De Pauw M, et al. Pharmacokinetics of multiple-dose darunavir in combination with low-dose ritonavir in individuals with mild-to-moderate hepatic impairment. Clin Pharmacokinet. 2010;49(5):343–50.

    Article  CAS  PubMed  Google Scholar 

  17. Rokitta D, Pfeiffer K, Streich C, Gerwin H, Fuhr U. The effect of organic solvents on enzyme kinetic parameters of human CYP3A4 and CYP1A2 in vitro. Toxicol Mech Methods. 2013;23(8):576–83.

    Article  CAS  PubMed  Google Scholar 

  18. Je YT, Sim WC, Kim DG, Jung BH, Shin HS, Lee BH. Expression of CYP3A in chronic ethanol-fed mice is mediated by endogenous pregnane X receptor ligands formed by enhanced cholesterol metabolism. Arch Toxicol. 2015;89(4):579–89.

    Article  CAS  PubMed  Google Scholar 

  19. Pasternak AO, Lukashov VV, Berkhout B. Cell-associated HIV RNA: a dynamic biomarker of viral persistence. Retrovirology. 2013;10:41.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Midde NM, Sinha N, Lukka PB, Meibohm B, Kumar S. Alterations in cellular pharmacokinetics and pharmacodynamics of elvitegravir in response to ethanol exposure in HIV-1 infected monocytic (U1) cells. PLoS One. 2017;12(2):e0172628.

    Article  PubMed  PubMed Central  Google Scholar 

  21. Mastrogiannis DS, Wang X, Dai M, Li J, Wang Y, Zhou Y, et al. Alcohol enhances HIV infection of cord blood monocyte-derived macrophages. Curr HIV Res. 2014;12(4):301–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Agudelo M, Figueroa G, Yndart A, Casteleiro G, Munoz K, Samikkannu T, et al. Alcohol and cannabinoids differentially affect HIV infection and function of human monocyte-derived dendritic cells (MDDC). Front Microbiol. 2015;6:1452.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Ande A, Sinha N, Rao PSS, McArthur CP, Ayuk L, Achu PN, et al. Enhanced oxidative stress by alcohol use in HIV+ patients: possible involvement of cytochrome P450 2E1 and antioxidant enzymes. AIDS Res Ther. 2015;12:29.

    Article  PubMed  PubMed Central  Google Scholar 

  24. DrugsFDA. http://www.accessdata.fda.gov/drugsatfda_docs/nda/2006/021976s000_Sprycel_ClinPharmR.pdf. [Accessed on 2016, January 7]. 2015.

  25. Boffito M, Jackson A, Amara A, Back D, Khoo S, Higgs C, et al. Pharmacokinetics of once-daily darunavir-ritonavir and atazanavir-ritonavir over 72 hours following drug cessation. Antimicrob Agents Chemother. 2011;55(9):4218–23.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Asai H, Imaoka S, Kuroki T, Monna T, Funae Y. Microsomal ethanol oxidizing system activity by human hepatic cytochrome P450s. J Pharmacol Exp Ther. 1996;277(2):1004–9.

    CAS  PubMed  Google Scholar 

  27. Hamitouche S, Poupon J, Dreano Y, Amet Y, Lucas D. Ethanol oxidation into acetaldehyde by 16 recombinant human cytochrome P450 isoforms: role of CYP2C isoforms in human liver microsomes. Toxicol Lett. 2006;167(3):221–30.

    Article  CAS  PubMed  Google Scholar 

  28. Cardoso JL, Lanchote VL, Pereira MP, Capela JM, de Moraes NV, Lepera JS. Impact of inhalational exposure to ethanol fuel on the pharmacokinetics of verapamil, ibuprofen and fluoxetine as in vivo probe drugs for CYP3A, CYP2C and CYP2D in rats. Food Chem Toxicol. 2015;84:99–105.

    Article  CAS  PubMed  Google Scholar 

  29. Salmela KS, Kessova IG, Tsyrlov IB, Lieber CS. Respective roles of human cytochrome P-4502E1, 1A2, and 3A4 in the hepatic microsomal ethanol oxidizing system. Alcohol Clin Exp Res. 1998;22(9):2125–32.

    Article  CAS  PubMed  Google Scholar 

  30. Sellers EM, Holloway MR. Drug kinetics and alcohol ingestion. Clin Pharmacokinet. 1978;3(6):440–52.

    Article  CAS  PubMed  Google Scholar 

  31. Kostrubsky VE, Strom SC, Wood SG, Wrighton SA, Sinclair PR, Sinclair JF. Ethanol and isopentanol increase CYP3A and CYP2E in primary cultures of human hepatocytes. Arch Biochem Biophys. 1995;322(2):516–20.

    Article  CAS  PubMed  Google Scholar 

  32. Feierman DE, Melinkov Z, Nanji AA. Induction of CYP3A by ethanol in multiple in vitro and in vivo models. Alcohol Clin Exp Res. 2003;27(6):981–8.

    Article  CAS  PubMed  Google Scholar 

  33. Zhou SF. Drugs behave as substrates, inhibitors and inducers of human cytochrome P450 3A4. Curr Drug Metab. 2008;9(4):310–22.

    Article  CAS  PubMed  Google Scholar 

  34. Lane EA, Guthrie S, Linnoila M. Effects of ethanol on drug and metabolite pharmacokinetics. Clin Pharmacokinet. 1985;10(3):228–47.

    Article  CAS  PubMed  Google Scholar 

  35. Szabo G, Mandrekar P. Human monocytes, macrophages, and dendritic cells: alcohol treatment methods. Methods Mol Biol. 2008;447:113–24.

    Article  CAS  PubMed  Google Scholar 

  36. Jin M, Ande A, Kumar A, Kumar S. Regulation of cytochrome P450 2e1 expression by ethanol: role of oxidative stress-mediated pkc/jnk/sp1 pathway. Cell Death Dis. 2013;4:e554.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Jin M, Earla R, Shah A, Earla RL, Gupte R, Mitra AK, et al. A LC-MS/MS method for concurrent determination of nicotine metabolites and role of CYP2A6 in nicotine metabolism in U937 macrophages: implications in oxidative stress in HIV + smokers. J NeuroImmune Pharmacol. 2012;7(1):289–99.

    Article  PubMed  Google Scholar 

  38. Zhou S-F, Xue CC, Yu X-Q, Li C, Wang G. Clinically important drug interactions potentially involving mechanism-based inhibition of cytochrome P450 3A4 and the role of therapeutic drug monitoring. Ther Drug Monit. 2007;29(6):687–710.

    Article  CAS  PubMed  Google Scholar 

  39. Kirby BJ, Collier AC, Kharasch ED, Whittington D, Thummel KE, Unadkat JD. Complex drug interactions of HIV protease inhibitors 1: inactivation, induction, and inhibition of cytochrome P450 3A by ritonavir or nelfinavir. Drug Metab Dispos: Biolo Fate Chem. 2011;39(6):1070–8.

    Article  CAS  Google Scholar 

  40. Vermeir M, Lachau-Durand S, Mannens G, Cuyckens F, van Hoof B, Raoof A. Absorption, metabolism, and excretion of darunavir, a new protease inhibitor, administered alone and with low-dose ritonavir in healthy subjects. Drug Metab Dispos: Biolo Fate Chem. 2009;37(4):809–20.

    Article  CAS  Google Scholar 

  41. Pasala S, Barr T, Messaoudi I. Impact of alcohol abuse on the adaptive immune system. Alcohol Res. 2015;37(2):185–97.

    PubMed  PubMed Central  Google Scholar 

  42. Harrison NL, Skelly MJ, Grosserode EP, Lowes DC, Zeric T, Phister S, et al. Effects of acute alcohol on excitability in the CNS. Neuropharmacology. 2017.

  43. Kenworthy KE, Clarke SE, Andrews J, Houston JB. Multisite kinetic models for CYP3A4: simultaneous activation and inhibition of diazepam and testosterone metabolism. Drug Metab Dispos. 2001;29(12):1644–51.

    CAS  PubMed  Google Scholar 

  44. Hosea NA, Miller GP, Guengerich FP. Elucidation of distinct ligand binding sites for cytochrome P450 3A4. Biochemistry. 2000;39(20):5929–39.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments and Disclosures

This research was supported by grants from the National Institute of Health to Santosh Kumar (NIAAA/NIH AA-022063) and Bernd Meibohm (OD/NIH S10OD016226), and from the National Natural Science Foundation of China to Weihua Li (Grant 81,373,328).

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Correspondence to Narasimha M. Midde or Santosh Kumar.

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Midde, N.M., Gong, Y., Cory, T.J. et al. Influence of Ethanol on Darunavir Hepatic Clearance and Intracellular PK/PD in HIV-Infected Monocytes, and CYP3A4-Darunavir Interactions Using Inhibition and in Silico Binding Studies. Pharm Res 34, 1925–1933 (2017). https://doi.org/10.1007/s11095-017-2203-6

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  • DOI: https://doi.org/10.1007/s11095-017-2203-6

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