Vaginal Gel Formulation Based on Theaflavin Derivatives As a Microbicide to Prevent HIV Sexual Transmission
Publication: AIDS Research and Human Retroviruses
Volume 28, Issue Number 11
Abstract
We previously demonstrated that a commercially available natural product preparation with high content (>90%) of theaflavin derivatives (TFmix) exhibited potent anti-HIV activities. Here we developed a TFmix gel formulation as a topical microbicide candidate. The effect of TFmix on the amyloid fibril formation of semen-derived enhancer of virus infection (SEVI) peptide was detected by transmission electron microscopy. The toxicity of the TFmix gel was evaluated using human vaginal and cervical epithelial cell lines and rabbit vaginal irritation models, respectively. Levels of proinflammatory cytokines (IL-1β, IL-6, IL-8, and TNF-α) and immunoregulatory cytokines (IL-10 and GM-CSF) in cervicovaginal lavages (CVLs) were measured by ELISA kits. Proliferating cell nuclear antigen (PCNA) immunostaining was performed to evaluate inflammation in the vaginal tissues. TFmix gel could degrade SEVI-specific amyloid fibrils and showed low cytotoxicity to epithelial cells of the female reproductive tract. No apparent cervicovaginal toxicity was observed at any time point evaluated following the intravaginal administration of TFmix gel to rabbits, whereas application of N-9 gel resulted in damage to the vaginal epithelium. Neither proinflammatory nor immunoregulatory cytokine production was triggered by TFmix gel. Only low expression of PCNA was observed in vaginal tissues of TFmix gel-treated rabbits. The concentration of TFmix in plasma was very low (below the lower limit of quantitation) 1 h after a single vaginal administration of TFmix gel. However, TFmix was still detected in the cervicovaginal lavages (CVLs) 6 h after treatment, indicating that it could be retained in the vaginal cavity for a long period of time. With its potent anti-HIV-1 activity, marked stability at acidic condition, low mucosal toxicity, and lack of systemic absorption, TFmix gel can be considered as an inexpensive and safe microbicide candidate for the prevention of HIV sexual transmission.
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References
1.
WHO2011progress report on the global AIDS epidemichttp://www.who.int/hiv/pub/progress_report2011/en/. 1. WHO. 2011 progress report on the global AIDS epidemic. http://www.who.int/hiv/pub/progress_report2011/en/.
2.
Gisselquist DPotterat JJ. Heterosexual transmission of HIV in Africa: An empiric estimateInt J STD AIDS200314162-173. 2. Gisselquist D and Potterat JJ: Heterosexual transmission of HIV in Africa: An empiric estimate. Int J STD AIDS 2003;14:162–173.
3.
Malcolm KWoolfson DToner C et al. Vaginal microbicides for the prevention of HIV transmissionBiotechnol Genet Eng Rev20042181-121. 3. Malcolm K, Woolfson D, Toner C, et al.: Vaginal microbicides for the prevention of HIV transmission. Biotechnol Genet Eng Rev 2004;21:81–121.
4.
Stone AJiang S. Microbicides: Stopping HIV at the gateLancet2006368431-433. 4. Stone A and Jiang S: Microbicides: Stopping HIV at the gate. Lancet 2006;368:431–433.
5.
Ramachandran RShanmughavel P. Role of microbicides in the prevention of HIV and sexually transmitted diseases—a reviewCurr HIV Res20097279-286. 5. Ramachandran R and Shanmughavel P: Role of microbicides in the prevention of HIV and sexually transmitted diseases—a review. Curr HIV Res 2009;7:279–286.
6.
McCormack S. Vaginal microbicidesCurr Opin Infect Dis20021557-62. 6. McCormack S: Vaginal microbicides. Curr Opin Infect Dis 2002;15:57–62.
7.
Abdool Karim QAbdool Karim SFrohlich J et al. Effectiveness and safety of tenofovir gel, an antiretroviral microbicide, for the prevention of HIV infection in womenScience201032959961168-1174. 7. Abdool Karim Q, Abdool Karim S, Frohlich J, et al.: Effectiveness and safety of tenofovir gel, an antiretroviral microbicide, for the prevention of HIV infection in women. Science 2010;329(5996):1168–1174.
8.
Van DLChandeying VRamjee G et al. Safety of multiple daily applications of COL-1492, a nonoxynol-9 vaginal gel, among female sex workersCOL-1492 Phase II Study Group. AIDS20001485-88. 8. Van DL, Chandeying V, Ramjee G, et al.: Safety of multiple daily applications of COL-1492, a nonoxynol-9 vaginal gel, among female sex workers. COL-1492 Phase II Study Group. AIDS 2000;14:85–88.
9.
Fichorova RNTucker LDAnderson DJ. The molecular basis of nonoxynol-9-induced vaginal inflammation and its possible relevance to human immunodeficiency virus type 1 transmissionJ Infect Dis2001184418-428. 9. Fichorova RN, Tucker LD, and Anderson DJ: The molecular basis of nonoxynol-9-induced vaginal inflammation and its possible relevance to human immunodeficiency virus type 1 transmission. J Infect Dis 2001;184:418–428.
10.
Van DLGovinden RMirembe FM et al. Lack of effectiveness of cellulose sulfate gel for the prevention of vaginal HIV transmissionN Engl J Med2008359463-472. 10. Van DL, Govinden R, Mirembe FM, et al.: Lack of effectiveness of cellulose sulfate gel for the prevention of vaginal HIV transmission. N Engl J Med 2008;359:463–472.
11.
Fichorova RNHidemi S et al. Novel vaginal microflora colonization model providing new insight into microbicide mechanism of actionMBio20112526e00168-11. 11. Fichorova RN, Hidemi S, et al.: Novel vaginal microflora colonization model providing new insight into microbicide mechanism of action. MBio 2011;25;2(6):e00168-11.
12.
Nath AGarg S. Microbicides in India-present and futureIndian J Med Microbiol200927251-253. 12. Nath A and Garg S: Microbicides in India-present and future. Indian J Med Microbiol 2009;27:251–253.
13.
Friedman M. Overview of antibacterial, antitoxin, antiviral, and antifungal activities of tea flavonoids and teasMol Nutr Food Res200751116-134. 13. Friedman M: Overview of antibacterial, antitoxin, antiviral, and antifungal activities of tea flavonoids and teas. Mol Nutr Food Res 2007;51:116–134.
14.
Ciraj AMSulaim JMamatha B et al. Antibacterial activity of black tea (Camelia sinensis) extract against Salmonella serotypes causing enteric feverIndian J Med Sci200155376-381. 14. Ciraj AM, Sulaim J, Mamatha B, et al.: Antibacterial activity of black tea (Camelia sinensis) extract against Salmonella serotypes causing enteric fever. Indian J Med Sci 2001;55:376–381.
15.
Lee HHHo CTLin JK. Theaflavin-3,3'-digallate and penta-O-galloyl-beta-D-glucose inhibit rat liver microsomal 5alpha-reductase activity and the expression of androgen receptor in LNCaP prostate cancer cellsCarcinogenesis2004251109-1118. 15. Lee HH, Ho CT, and Lin JK: Theaflavin-3,3'-digallate and penta-O-galloyl-beta-D-glucose inhibit rat liver microsomal 5alpha-reductase activity and the expression of androgen receptor in LNCaP prostate cancer cells. Carcinogenesis 2004;25:1109–1118.
16.
Tu YYTang ABWatanabe N. The theaflavin monomers inhibit the cancer cells growth in vitroActa Biochim Biophys Sin (Shanghai)200436508-512. 16. Tu YY, Tang AB, and Watanabe N: The theaflavin monomers inhibit the cancer cells growth in vitro. Acta Biochim Biophys Sin (Shanghai) 2004;36:508–512.
17.
Aneja ROdoms KDenenberg AG et al. Theaflavin, a black tea extract, is a novel anti-inflammatory compoundCrit Care Med2004322097-2103. 17. Aneja R, Odoms K, Denenberg AG, et al.: Theaflavin, a black tea extract, is a novel anti-inflammatory compound. Crit Care Med 2004;32:2097–2103.
18.
Huang MTLiu YRamji D et al. Inhibitory effects of black tea theaflavin derivatives on 12-O-tetradecanoylphorbol-13-acetate-induced inflammation and arachidonic acid metabolism in mouse earsMol Nutr Food Res200650115-122. 18. Huang MT, Liu Y, Ramji D, et al.: Inhibitory effects of black tea theaflavin derivatives on 12-O-tetradecanoylphorbol-13-acetate-induced inflammation and arachidonic acid metabolism in mouse ears. Mol Nutr Food Res 2006;50:115–122.
19.
Sang SLambert JDTian S et al. Enzymatic synthesis of tea theaflavin derivatives and their anti-inflammatory and cytotoxic activitiesBioorg Med Chem200412459-467. 19. Sang S, Lambert JD, Tian S, et al.: Enzymatic synthesis of tea theaflavin derivatives and their anti-inflammatory and cytotoxic activities. Bioorg Med Chem 2004;12:459–467.
20.
Cai FLi CRWu JL et al. Theaflavin ameliorates cerebral ischemia- reperfusion injury in rats through its anti-inflammatory effect, modulation of STAT-1Mediators Inflamm2006530490. 20. Cai F, Li CR, Wu JL, et al.: Theaflavin ameliorates cerebral ischemia- reperfusion injury in rats through its anti-inflammatory effect and modulation of STAT-1. Mediators Inflamm 2006(5);30490.
21.
Letchoumy PVMohan KVStegeman JJ et al. In vitro antioxidative potential of lactoferrin and black tea polyphenols and protective effects in vivo on carcinogen activation, DNA damage, proliferation, invasion, and angiogenesis during experimental oral carcinogenesisOncol Res200817193-203. 21. Letchoumy PV, Mohan KV, Stegeman JJ, et al.: In vitro antioxidative potential of lactoferrin and black tea polyphenols and protective effects in vivo on carcinogen activation, DNA damage, proliferation, invasion, and angiogenesis during experimental oral carcinogenesis. Oncol Res 2008;17:193–203.
22.
Yoshino KHara YSano M et al. Antioxidative effects of black tea TF and thearubigin on lipid peroxidation of rat liver homogenates induced by tert-butyl hydroperoxideBiol Pharm Bull199417146-149. 22. Yoshino K, Hara Y, Sano M, et al.: Antioxidative effects of black tea TF and thearubigin on lipid peroxidation of rat liver homogenates induced by tert-butyl hydroperoxide. Biol Pharm Bull 1994;17:146–149.
23.
Liang YCChen YCLin YL et al. Suppression of extracellular signals and cell proliferation by the black tea polyphenol, theaflavin-3,3'-digallateCarcinogenesis199920733-736. 23. Liang YC, Chen YC, Lin YL, et al.: Suppression of extracellular signals and cell proliferation by the black tea polyphenol, theaflavin-3,3'-digallate. Carcinogenesis 1999;20:733–736.
24.
Kundu TDey SRoy M et al. Induction of apoptosis in human leukemia cells by black tea and its polyphenol theaflavinCancer Lett2005230111-121. 24. Kundu T, Dey S, Roy M, et al.: Induction of apoptosis in human leukemia cells by black tea and its polyphenol theaflavin. Cancer Lett 2005;230:111–121.
25.
Lahiry LSaha BChakraborty J et al. Contribution of p53-mediated Bax transactivation in theaflavin-induced mammary epithelial carcinoma cell apoptosisApoptosis200813771-781. 25. Lahiry L, Saha B, Chakraborty J, et al.: Contribution of p53-mediated Bax transactivation in theaflavin-induced mammary epithelial carcinoma cell apoptosis. Apoptosis 2008;13:771–781.
26.
Nance CLSiwak EBShearer WT. Preclinical development of the green tea catechin, epigallocatechin gallate, as an HIV-1 therapyJ Allergy Clin Immunol2009123459-465. 26. Nance CL, Siwak EB, and Shearer WT: Preclinical development of the green tea catechin, epigallocatechin gallate, as an HIV-1 therapy. J Allergy Clin Immunol 2009;123:459–465.
27.
Williamson MPMcCormick TGNance CL et al. Epigallocatechin gallate, the main polyphenol in green tea, binds to the T-cell receptor, CD4: Potential for HIV-1 therapyJ Allergy Clin Immunol20061181369-1374. 27. Williamson MP, McCormick TG, Nance CL, et al.: Epigallocatechin gallate, the main polyphenol in green tea, binds to the T-cell receptor, CD4: Potential for HIV-1 therapy. J Allergy Clin Immunol 2006;118:1369–1374.
28.
Nance CLShearer WT. Is green tea good for HIV-1 infection?J Allergy Clin Immunol2003112851-853. 28. Nance CL and Shearer WT: Is green tea good for HIV-1 infection?. J Allergy Clin Immunol 2003;112:851–853.
29.
Liu SLu HZhao Q et al. Theaflavin derivatives in black tea and catechin derivatives in green tea inhibit HIV-1 entry by targeting gp41Biochim Biophys Acta20051723270-281. 29. Liu S, Lu H, Zhao Q, et al.: Theaflavin derivatives in black tea and catechin derivatives in green tea inhibit HIV-1 entry by targeting gp41. Biochim Biophys Acta 2005;1723:270–281.
30.
Yang JLi LTan SY et al. A natural theaflavins preparation inhibits HIV-1 infection by targeting the entry step: Potential applications for preventing HIV-1 infectionFitoterapia201283348-355. 30. Yang J, Li L, Tan SY, et al.. A natural theaflavins preparation inhibits HIV-1 infection by targeting the entry step: Potential applications for preventing HIV-1 infection. Fitoterapia 2012;83:348–355.
31.
Wang KLiu ZHuang J et al. Determination of TF in black tea by high performance liquid chromatographySe Pu200422151-153. 31. Wang K, Liu Z, Huang J, et al.: Determination of TF in black tea by high performance liquid chromatography. Se Pu 2004;22:151–153.
32.
Owen DHKatz DF. A vaginal fluid simulantContraception19995991-95. 32. Owen DH and Katz DF: A vaginal fluid simulant. Contraception 1999;59:91–95.
33.
Hauber IHohenberg HHolstermann B et al. The main green tea polyphenol epigallocatechin-3-gallate counteracts semen-mediated enhancement of HIV infectionProc Natl Acad Sci USA20091069033-9038. 33. Hauber I, Hohenberg H, Holstermann B, et al.: The main green tea polyphenol epigallocatechin-3-gallate counteracts semen-mediated enhancement of HIV infection. Proc Natl Acad Sci USA 2009;106:9033–9038.
34.
Tipton DALyle BBabich H et al. In vitro cytotoxic and anti-inflammatory effects of myrrh oil on human gingival fibroblasts and epithelial cellsToxicol In Vitro200317301-310. 34. Tipton DA, Lyle B, Babich H, et al.: In vitro cytotoxic and anti-inflammatory effects of myrrh oil on human gingival fibroblasts and epithelial cells. Toxicol In Vitro 2003;17:301–310.
35.
Fichorova RNRheinwald JGAnderson DJ. Generation of papillomavirus-immortalized cell lines from normal human ectocervical, endocervical, and vaginal epithelium that maintain expression of tissue-specific differentiation proteinsBiol Reprod1997574847-855. 35. Fichorova RN, Rheinwald JG, and Anderson DJ: Generation of papillomavirus-immortalized cell lines from normal human ectocervical, endocervical, and vaginal epithelium that maintain expression of tissue-specific differentiation proteins. Biol Reprod 1997;57(4):847–855.
36.
Fichorova RNAnderson DJ. Differential expression of immunobiological mediators by immortalized human cervical and vaginal epithelial cellsBiol Reprod1999602508-514. 36. Fichorova RN and Anderson DJ: Differential expression of immunobiological mediators by immortalized human cervical and vaginal epithelial cells. Biol Reprod 1999;60(2):508–514.
37.
Chou TCHayball MPCalcuSyn: Windows software for dose effect analysisBIOSOFTFerguson, MO1991. 37. Chou TC and Hayball MP: CalcuSyn: Windows software for dose effect analysis. BIOSOFT, Ferguson, MO, 1991.
38.
Aranha CCGupta SMReddy KV. Assessment of cervicovaginal cytokine levels following exposure to microbicide Nisin gel in rabbitsCytokine20084363-70. 38. Aranha CC, Gupta SM, and Reddy KV: Assessment of cervicovaginal cytokine levels following exposure to microbicide Nisin gel in rabbits. Cytokine 2008;43:63–70.
39.
Draize JHAppraisal of the safety of chemicals in foods, drugs and cosmeticsUnited StatesAssociation of Food and Drug OfficialsDermal Toxicity195946-59. 39. Draize JH: Appraisal of the safety of chemicals in foods, drugs and cosmetics. United States: Association of Food and Drug Officials. Dermal Toxicity 1959;46–59.
40.
D'Cruz OJSamuel PWaurzyniak B et al. Development and evaluation of a thermoreversible ovule formulation of stampidine, a novel nonspermicidal broad-spectrum anti-human immunodeficiency virus microbicideBiol Reprod2003691843-1851. 40. D'Cruz OJ, Samuel P, Waurzyniak B, et al.: Development and evaluation of a thermoreversible ovule formulation of stampidine, a novel nonspermicidal broad-spectrum anti-human immunodeficiency virus microbicide. Biol Reprod 2003;69:1843–1851.
41.
Eckstein PJackson MCMillman N et al. Comparison of vaginal tolerance tests of spermicidal preparations in rabbits and monkeysJ Reprod Fertil19692085-93. 41. Eckstein P, Jackson MC, Millman N, et al.: Comparison of vaginal tolerance tests of spermicidal preparations in rabbits and monkeys. J Reprod Fertil 1969;20:85–93.
42.
Costa PLobo JM. Modeling and comparison of dissolution profilesEur J Pharm Sci200113123-133. 42. Costa P and Lobo JM. Modeling and comparison of dissolution profiles. Eur J Pharm Sci 2001;13:123–133.
43.
Munch JRucker EStandker L et al. Semen-derived amyloid fibrils drastically enhance HIV infectionCell20071311059-1071. 43. Munch J, Rucker E, Standker L, et al.: Semen-derived amyloid fibrils drastically enhance HIV infection. Cell 2007;131:1059–1071.
44.
Yu DMorris-Natschke SLLee KH. New developments in natural products-based anti-AIDS researchMed Res Rev200727108-132. 44. Yu D, Morris-Natschke SL, and Lee KH: New developments in natural products-based anti-AIDS research. Med Res Rev 2007;27:108–132.
45.
Dufresne CJFarnworth ER. A review of latest research findings on the health promotion properties of teaJ Nutr Biochem200112404-421. 45. Dufresne CJ and Farnworth ER: A review of latest research findings on the health promotion properties of tea. J Nutr Biochem 2001;12:404–421.
46.
Hussar DACampoli KE. New drugs: Etravirine, sinecatechins, and desvenlafaxine succinateJ Am Pharm Assoc200848428-431. 46. Hussar DA and Campoli KE: New drugs: Etravirine, sinecatechins, and desvenlafaxine succinate. J Am Pharm Assoc 2008;48:428–431.
47.
Tatti SSwinehart JMThielert C et al. Sinecatechins, a defined green tea extract, in the treatment of external anogenital warts: A randomized controlled trialObstet Gynecol20081111371-1379. 47. Tatti S, Swinehart JM, Thielert C, et al.: Sinecatechins, a defined green tea extract, in the treatment of external anogenital warts: A randomized controlled trial. Obstet Gynecol 2008;111:1371–1379.
48.
Jiang SDebnath AK. Development of HIV entry inhibitors targeted to the coiled-coil regions of gp41Biochem Biophys Res Commun2000269641-646. 48. Jiang S and Debnath AK: Development of HIV entry inhibitors targeted to the coiled-coil regions of gp41. Biochem Biophys Res Commun 2000;269:641–646.
49.
Lederman MMJump RPilch-Cooper HA et al. Topical application of entry inhibitors as "virustats" to prevent sexual transmission of HIV infectionRetrovirology20085116. 49. Lederman MM, Jump R, Pilch-Cooper HA, et al.: Topical application of entry inhibitors as "virustats" to prevent sexual transmission of HIV infection. Retrovirology 2008;5:116.
50.
Rabiskova MSedlakova MVitkova M et al. Carbomers and their use in pharmaceutical technologyCeska Slov Farm200453300-303. 50. Rabiskova M, Sedlakova M, Vitkova M, et al.: Carbomers and their use in pharmaceutical technology. Ceska Slov Farm 2004;53:300–303.
51.
Fichorova RNBajpai MChandra N et al. Interleukin (IL)-1, IL-6, and IL-8 predict mucosal toxicity of vaginal microbicidal contraceptivesBiol Reprod200471761-769. 51. Fichorova RN, Bajpai M, Chandra N, et al.: Interleukin (IL)-1, IL-6, and IL-8 predict mucosal toxicity of vaginal microbicidal contraceptives. Biol Reprod 2004;71:761–769.
52.
Fichorova RN. Guiding the vaginal microbicide trials with biomarkers of inflammationJ Acquir Immune Defic Syndr200437(Suppl)3184-193. 52. Fichorova RN: Guiding the vaginal microbicide trials with biomarkers of inflammation. J Acquir Immune Defic Syndr 2004; 37(Suppl)3:184–193.
Information & Authors
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Published In

AIDS Research and Human Retroviruses
Volume 28 • Issue Number 11 • November 2012
Pages: 1498 - 1508
PubMed: 22867271
Copyright
Copyright 2012, Mary Ann Liebert, Inc.
History
Published online: 5 November 2012
Published in print: November 2012
Published ahead of print: 11 September 2012
Published ahead of production: 6 August 2012
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No competing financial interests exist.
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