HIV-1 REVERSE TRANSCRIPTASE AND PROTEASE ASSAY OF METHANOLIC EXTRACTS OF ADANSONIA DIGITATA L.
Objective: Anti-HIV activity studies of methanolic extracts of Adansonia digitata L. leaves, root bark and fruit pulp.
Methods: Methanolic extracts of A. digitata were tested for HIV enzyme inhibitory activity against HIV-1 reverse transcriptase (RT) and HIV-1 protease (PR). HIV-1 RT assay was performed using non-radioactive HIV-RT colorimetric ELISA kit while the HIV-1 PR assay was performed using a fluorogenic octapeptide substrate, HIV-FRET (fluorescence resonance energy transfer) and a recombinant HIV-1 protease solution (AnaSpec Inc., USA).
Results: The percentage inhibition of controls and A. digitata leaves, root bark and fruit pulp extracts were calculated relative to uninhibited HIV-1 RT and PR in 2% DMSO. The results of the HIV-1 RT assay indicated 26.5% inhibition by root bark extract (50 Âµg/ml) and 12.2% inhibition by leaf extract while the fruit pulp extract (100 Âµg/ml) has shown only 5.9% inhibition as compared to standard Nevirapine (100 nM) 27.5% inhibition.
In the case of HIV-protease assay, leaf extract and fruit pulp extract (50 Âµg/ml) has shown quite high inhibition (â‰¥ 50%) to the extent of 75% and 74% respectively which is very significant. The root bark extract has shown 35% inhibition.
Conclusion: Methanolic extract of the leaves, root bark and fruit pulp of A. digitata has shown low anti-HIV-1 RT but significant PR activity. HIV-1 RT activity of root bark extract and HIV-1 PR activity of fruit pulp extract indicates the potential of the plant as an anti-HIV agent.Â
2. Fletcher CV, Kakuda TN, Collier AC, Talbert RL, Yee GC, Matzke GR, et al. Pharmacotherapy-a pathophysiologic approach. 5th edition. Mcgraw-Hill: 2002. Medical Publishing Division, United States of America; 2002. p. 2151-74.
3. Cos P, Maes L, Vanden Berghe D, Hermans N, Pieters L, Vlietinck A. Plant substances as anti-HIV agents selected according to their putative mechanism of action. J Nat Prod 2004;67:284-93.
4. Mandal D, Jana S, Bhattacharya SK, Chakrabarti S. HIV type 1 subtypes circulating in eastern and northeastern regions of India. AIDS Res Hum Retroviruses 2002;18:1219-27.
5. National AIDS Control Organization (NACO). National AIDS Control Organisation 2006. Available from http:// www.nacoonline.org. [Last accessed on 20 Mar 2016].
6. De Clercq E. Current lead natural products for the chemotherapy of human immunodeficiency virus (HIV) infection. Med Res Rev 2000;20:323-49.
7. Cos P, Vanden Berghe D, Bruyne TD, Vlietinck A. Plant substances as anti-viral agents: an update (1997â€“2001). Curr Org Chem 2004;7:1163-80.
8. Harnett SM, Oosthuizen V, van de Venter M. Anti-HIV activities of organic and aqueous extracts of Sutherlandia frutescens and Lobostemon trigonus. J Ethnopharmcol 2005;96:113-9.
9. Zhang K, Wang J, Jiang Y, Xu LZ. Utilization of traditional Chinese medicines in the treatment of 200 people living with HIV/AIDS. Chin J AIDS STD 2005;11:94-6.
10. Asres K, Bucar F, Kartnig T, Witvrouw M, Pannecouque C, De Clercq E. Antiviral activity against human immunodeficiency virus type 1 (HIV-1) and type 2 (HIV-2) of ethnobotanically selected ethiopian medicinal plants. Phytother Res 2001;15:62-9.
11. Vermani K, Garg S. Herbal medicines for sexually transmitted diseases and AIDS. J Ethnopharmacol 2002;80:49-66.
12. Farnsworth NR. Ethnopharmacology and drug development. Ethnobotany, drug development and biodiversity conservation-exploring the linkages. In: Ethnobotany and the Search for New Drugs; Ciba Foundation Symposium. John Wiley and Sons, Chichester; 1995;185:42-59.
13. Fabricant DS, Farnsworth NR. The value of plants used in traditional medicine for drug discovery. Environ Health Perspect 2001;109:69-75.
14. Wang JH, Tam SC, Huang H, Ouyang DY, Wang YY, Zheng YT. Site-directed PEGylation of trichosanthin retained its anti-HIV activity with reduces potency in vitro. Biochem Biophys Res Commun 2004a;317:965-71.
15. Ng TB, Huang B, Fong WP, Yeung HW. Anti-human immunodeficiency virus (anti-HIV) natural products with special emphasis on HIV reverse transcriptase inhibitors. Life Sci 1997;61:933-49.
16. Vlietinck AJ, De Bruyne T, Apers S, Pieters LA. Plant-derived leading compounds for chemotherapy of human immunodeficiency virus (HIV) infection. Planta Med 1998;64:97-109.
17. Yang SS, Gragg GM, Newman DJ, Bader JP. Natural product based anti-HIV drug discovery and development facilitated by the NCI developmental therapeutics program. J Nat Prod 2001;64:265-77.
18. Rangari VD, Dumbre RK, Dumbre MR. HIV-AIDS and bioactive natural products. Studium Press Llc, Houston, Texas, USA; 2009.
19. Nadkarni KM, Nadkarni AK. Indian Materia Medica-2. 3rd ed. Popular Prakasan, Bombay; 2000. p. 37-9.
20. Ramadan A, Harraz FM, El-Mougy SA. Anti-inflammatory, analgesic and antipyretic effects of the fruit pulp of Adansonia digitata. Fitoterapia 1994;LXV:418â€“22.
21. Shukla YN, Dubey S, Jain SP, Kumar S. Chemistry, biology and uses of Adansonia digitata-a review. J Med Aromat Plant Sci 2001;23:429-34.
22. Ramesh D, Dennis TJ, Shingare MS. Constituents of Adansoia digitata root bark. Fitoterapia 1992;63:278-9.
23. Van wyk BE, Gericke N. Peopleâ€™s Plants: a guide to useful plants of Southern Africa. Pretoria, Briza Publication; 2000. p. 351.
24. Esterhuyse N, Von Breitenbach J, Suhnge H. Remarkable trees of South Africa. Pretoria, Briza Publication; 2001.
25. Wickens GE. The Baobab-Africaâ€™s upside-down tree. Kew Bull 1982;37:173-209.
26. Van wyk BE, Van Heerden F, Van Oudtshoorn B. Poisonous plants of South Africa. Pretoria, Briza Publication; 2002. p. 288.
27. Hudson JB, Anani K, Lee MX, De SouzÃ¢ C, Arnason JT, Gbeassor M. Further investigations on the antiviral activities of medicinal plants of Togo. Pharm Biol 2000;38:46-50.
28. Atawodi SE, Bulus T, Ibrahim S, Amed DA, Nok AJ, Mamman M, Galadima M. In vitro trypanocidal effect of methanolic extract of some Nigerian savannah plants. Afr J Biotechnol 2003;2:317-21.
29. Kamatou GPP, Vermaak I, Viljoen AM. An updated review of Adansonia digitata: a commercially important African tree. S Afr J Bot 2011;77:908-19.
30. Anani K, Hudson JB, de Souzal C, Akpaganal K, Tower GHN, Amason, et al. Investigation of medicinal plants of Togo for antiviral and antimicrobial activities. Pharm Biol 2000;38:40-5.
31. Matayoshi ED, Wang GT, Krafft GA, Erickson J. Novel fluorogenic substrates for assaying retroviral proteases by resonance energy transfer. Science 1990;247:954-8.
32. Sharma A, Rangari VD. Antibacterial, the antifungal and antitubercular activity of methanolic extracts of Adansonia digitata. Int J Pharm Biol Sci 2015;10:52-60.
33. Chauhan JS, Chaturvedi R, Kumar S. A new Flavonol; l glycoside from the roots of Adansonia digitata. Planta Med 1984;50:113.
34. Chauhan JS, Kumar S, Chaturvedi R. A new flavonone glycoside from the roots of Adansonia digitata. Natl Acad Sci Lett 1987;10:177-9.
35. Yasmina A Lim, Ma Chao Mei, Ines T Kusumoto, Miyashiro H, Hattori M. HIV-1 Reverse transcriptase inhibitory principles from Chamaesyce hyssopifolia. J Phytother Res 1997;11:22-7.
36. Kannan M, Rajendran P, Vedha V, Ashok G, Anushka S, Chandran, et al. HIV-1 reverse transcriptase inhibition by Vitex negundo L. leaf extract and quantification of flavonoids in relation to the anti-HIV activity. J Cell Mol Biol 2012;10:53-9.
37. Shahat AA. Procyanidins from Adansonia digitata. Pharm Biol 2006;44:445-50.
38. Chaomei MA, Nakamura N, Miyashiro H, Hattori M, Shimotohno K. Inhibitory effect of constituents from Cynomorium songaricum and related triterpene derivatives on HIV-1 Protease. Chem Pharm Bull 1999;47:141-5.
39. Hong-XI XU, Wan M, Dong H, But PPH, Foo LY. Inhibitory activity of flavonoids and tannins against HIV-1 protease. Biol Pharm Bull 2000;23:1072-6.
40. Vlietinck AJ, Bruyne De T, Apers S, Pieters LA. Plant-derived leading compounds for chemotherapy of human immunodeficiency virus (HIV) infection. Planta Med 1998;64:97-109.