Amino Acid Conjugation: An Approach to Enhance Aqueous Solubility and Permeability of Poorly Water Soluble Drug Ritonavir
Abstract
Objective: The objective of present work is to improve physicochemical and pharmacokinetic profile of poorly water soluble HIV protease inhibitor, ritonavir (RT) by preparing its amino acid conjugates.
Methods: Ester conjugates of ritonavir with various amino acids were synthesized by a simple esterification process using dicyclohexyl carbodiimide (DCC) as a coupling agent. The synthesized compounds were characterized by thin layer chromatography (TLC), fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR) and mass spectroscopy. All conjugates were evaluated by saturation solubility and hydrolytic stability studies. Cytotoxicity and permeability studies were conducted using Caco-2 cell line.
Results: All amino acid conjugates showed a significantly higher aqueous solubility compared to the pure RT. With respect to hydrolysis, alkaline hydrolysis of conjugates was rapid relative to acidic hydrolysis. No cytotoxicity was shown by conjugates for concentration as high as 100μM, which indicates promising therapeutic potential. Transport studies across Caco-2 cells showed that prepared amino acid conjugates improved the permeation of RT compared to pure RT.
Conclusion: In vitro studies demonstrated that amino acid conjugation of RT may be an effective strategy to improve its aqueous solubility as well as permeability and can be used to improve oral absorption and thereby oral bioavailability of protease inhibitors.
Keywords: Ritonavir, amino acids, conjugates, solubility, HIV, hydrolysis
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References
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2. Liu H, Edgar KJ, Synthesis and characterization of neutral and anionic cellulosic amphiphiles, Carbohydrate Polymers, 2014; 113:480–489.
3. Liu H, Ilevbare GA, Cherniawski BP, et al. Synthesis and structure–property evaluation of cellulose x-carboxyesters for amorphous solid dispersions, Carbohydrate Polymers, 2014; 100:116–125.
4. Potta SG, Minemi S, Nukala RK, et al. Development of solid lipid nanoparticles for enhanced solubility of poorly soluble drugs, Journal of Biomedical Nanotechnology, 2010;6:634–640.
5. Rabinow BE, Nanosuspensions in drug delivery, Nature Reviews Drug Discovery, 2004; 3:785–796.
6. Muller RH, Peters K, Nanosuspensions for the formulation of poorly soluble drugs: I. Preparation by a size-reduction technique, International Journal of Pharmaceutics, 1998; 160:229–237.
7. Berge SM, Bighley LD, Monkhouse DC. Pharmaceutical salts, Journal of Pharmaceutical Sciences,1977; 66:1–19.
8. Fatouros D, Deen GR, Arleth L, et al. Structural development of self nano emulsifying drug delivery systems (SNEDDS) during in vitro lipid digestion monitored by small-angle X-ray scattering, Pharmaceutical Research, 2007; 24:1844–1853.
9. Lavasanifar A, Samuel J, Kwon GS, Poly(ethylene oxide)block-poly(l-amino acid) micelles for drug delivery, Advanced Drug Delivery Reviews, 2002; 54:169–190.
10. Loftsson T, Duchene D, Cyclodextrins and their pharmaceutical applications, International Journal of Pharmaceutics, 2007; 329:1–11.
11. Mueller E, Kovarik J, van Bree J, et al. Improved dose linearity of cyclosporine pharmacokinetics from a microemulsion formulation, Pharmaceutical Research, 1994; 11:301–304.
12. Park JH, Choi HK, Enhancement of solubility and dissolution of cilostazol by solid dispersion technique, Archives of Pharmacal Research, 2015; 38:1336–1344.
13. Saoji SD, Raut NA, Dhore PW, et al. Preparation and evaluation of phospholipid-based complex of standardized centella extract (SCE) for the enhanced delivery of phytoconstituents, AAPS Journal, 2016; 18:102–114.
14. Khan J, Saraf S, Saraf S, Preparation and evaluation of luteolin-phospholipid complex as an effective drug delivery tool against GalN/LPS induced liver damage, Pharmaeutical Development and Technology, 2016; 21:475–486
15. Daniela HJ, Diego EC, Man CC,et.al. The Prodrug Approach: A Successful Tool for Improving Drug Solubility, Molecules, 2016; 21, 42.
16. Zawilska J B, Wojcieszak J, Olejniczak AB, Prodrugs: A challenge for the drug development, Pharmacological Reports, 2013; 65:1–14.
17. Stella VJ, Prodrugs: Some thoughts and current issues, Journal of Pharmaceutical Sciences, 2010; 99:4755-65.
18. Huttunen KM, Raunio H, Rautio J, Prodrugs—from Serendipity to Rational Design. Pharmacological Reviews, 2011;63:750 –771.
19. Agarwal, S., Boddu, S.H., Jain, R.; Samanta, S.; Pal, D.; Mitra, A.K, Peptide prodrugs: Improved oral absorption of lopinavir, a HIV protease inhibitor, International Journal of Pharmaceutics, 2008; 359: 7–14.
20. Patel, M., Mandava, N., Gokulgandhi, M., Pal, D., Mitra, A.K, Amino Acid Prodrugs: An Approach to Improve the Absorption of HIV-1 Protease Inhibitor, Lopinavir, Pharmaceuticals,2014; 7:433-452.
21. Jain, R., Duvvuri, S., Kansara, V., Mandava, N.K., Mitra, A.K, Intestinal absorption of novel-dipeptide prodrugs of saquinavir in rats, International Journal of Pharmaceutics, 2007; 336:233–240.
22. Dias C, Nashed Y, Atluri H, Mitra A, Ocular penetration of acyclovir and its peptide prodrugs valacyclovir and val-valacyclovir following systemic administration in rabbits: An evaluation using ocular microdialysis and LC-MS, Current Eye Research, 2002; 25:243–252.
23. Patel K, Trivedi S, Luo S, Zhu X, Pal D, Kern ER, Mitra AK, Synthesis, physicochemical properties and antiviral activities of ester prodrugs of ganciclovir, International Journal of Pharmaceutics, 2005; 305:75–89.
24. Gunda S, Hariharan S, Mitra AK, Corneal absorption and anterior chamber pharmacokinetics of dipeptide monoester prodrugs of ganciclovir (GCV): In vivo comparative evaluation of these prodrugs with Val-GCV and GCV in rabbits, Journal of Ocular Pharmacology and Therapeutics, 2006; 22:465–476.
25. Majumdar S. et.al. Dipeptide monoester ganciclovir prodrugs for treating HSV-1-induced corneal epithelial and stromal keratitis: In vitro and in vivo evaluations, Journal of Ocular Pharmacology and Therapeutics, 2005; 21:463–474.
26. Gupta SV.et.al. Enhancing the intestinal membrane permeability of zanamivir: A carrier mediated prodrug approach, Molecular Pharmaceutics, 2011; 8:2358–2367.
27. Ghadi R, Dand N, BCS class IV drugs: Highly notorious candidates for formulation development, Journal of Controlled Release.2017; 28:71-95.
28. Law D, Schmitt EA, Marsh KC, Everitt EA, Wang W, Fort JJ, Krill SL, Qiu Y, Ritonavir-PEG 8000 amorphous solid dispersions: in vitro and in vivo evaluations. Journal of Pharmaceutical Sciences, 2004; 93(3):563-70.
29. Mahfouz NM, Hassan MA, Synthesis, chemical and enzymatic hydrolysis, and bioavailability evaluation in rabbits of metronidazole amino acid ester prodrugs with enhanced water solubility, Journal of Pharmacy and Pharmacology, 2001;53:841-48.
30. Xiangguo Z, Xinyi T, Dongzhi W, Qingxun S, Pharmacological activity and hydrolysis behavior of novel ibuprofen glucopyranoside conjugates, European Journal of Medicinal Chemistry, 2006; 41:1352-58.
31. Dhokchawle BV, Bhandari A, Synthesis, Hydrolysis Kinetics and Pharmacological Evaluation of Aceclofenac Prodrugs, Antiinflamm Antiallergy Agents in Medicinal Chemistry, 2014; 13:188-94.
32. Joshi G, Kumar A, Sawant K, Bioavailability enhancement, Caco-2 cells uptake and intestinal transport of orally administered lopinavir-loaded PLGA nanoparticles, Drug Delivery, 2016; 23(9): 3492-3504.
33. Senanayake TH, Warren G, Wei X, Vinogradov SV, Application of activated nucleoside analogs for the treatment of drug-resistant tumors by oral delivery of nanogel-drug conjugates, Journal of Controlled Release, 2003; 167:200–9.
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Chabukswar AR, Jagdale S, Gandhi P. Amino Acid Conjugation: An Approach to Enhance Aqueous Solubility and Permeability of Poorly Water Soluble Drug Ritonavir. JDDT [Internet]. 15May2019 [cited 2Jul.2020];9(3):252-6. Available from: http://jddtonline.info/index.php/jddt/article/view/2848
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