The gastrointestinal tract is structurally and functionally different from the vagina. Thus, the paradigm of topical microbicide development and evaluation has evolved to include rectal microbicides (RMs). Our interest was to create unique RM formulations to safely and effectively deliver antiretroviral drugs to mucosal tissue. RMs were designed to include those that spread and coat all surfaces of the rectum and distal colon rapidly (liquid) and those that create a deformable, erodible barrier and remain localized at the administration site (gel). Tenofovir (TFV) (1%) was formulated as an aqueous thermoreversible fluid and a carbopol-based aqueous hydrogel. Lipid-based liquid and gel formulations were prepared for UC781 (0.1%) using isopropyl myristate and GTCC (Caprylic/Capric Triglycerides), respectively. Formulations were characterized for pH, viscosity, osmolality, and drug content. Pre-clinical testing incorporated ex vivo colonic tissue obtained through surgical resections and flexible sigmoidoscopy (flex sig). As this was the first time using tissue from both sources side-by-side, the ability to replicate HIV-1 was compared. Efficacy of the RM formulations was tested by applying the products with HIV-1 directly to polarized colonic tissue and following viral replication. Safety of the formulations was determined by MTT assay and histology. All products had a neutral pH and were isoosmolar. While HIV-1BaL and HIV-1JR-CSF alone and in the presence of semen had similar replication trends between surgically resected and flex sig tissues, the magnitude of viral replication was significantly better in flex sig tissues. Both TFV and UC781 formulations protected the colonic tissue, regardless of tissue source, from HIV-1 and retained tissue viability and architecture. Our in vitro and ex vivo results show successful formulation of unique RMs. Moreover, the results of flex sig and surgically resected tissues were comparable suggesting the incorporation of both in pre-clinical testing algorithms.
References
[1]
WHO/UNAIDS/UNICEF (2011) Global HIV/AIDS Response. Geneva: UNAIDS.
[2]
Stein ZA (1990) HIV prevention: the need for methods women can use. Am J Public Health 80: 460–462. doi: 10.2105/ajph.80.4.460
[3]
Beyrer C, Baral SD, van Griensven F, Goodreau SM, Chariyalertsak S, et al. (2012) Global epidemiology of HIV infection in men who have sex with men. Lancet 380: 367–377. doi: 10.1016/s0140-6736(12)60821-6
[4]
El-Sadr WM, Mayer KH, Hodder SL (2010) AIDS in America–forgotten but not gone. N Engl J Med 362: 967–970. doi: 10.1056/nejmp1000069
[5]
Beyrer C (2010) Global prevention of HIV infection for neglected populations: men who have sex with men. Clin Infect Dis 50 Suppl 3S108–113. doi: 10.1086/651481
[6]
Gorbach PM, Manhart LE, Hess KL, Stoner BP, Martin DH, et al. (2009) Anal intercourse among young heterosexuals in three sexually transmitted disease clinics in the United States. Sex Transm Dis 36: 193–198. doi: 10.1097/olq.0b013e3181901ccf
[7]
Herbenick D, Reece M, Schick V, Sanders SA, Dodge B, et al. (2010) Sexual behavior in the United States: results from a national probability sample of men and women ages 14–94. J Sex Med 7 Suppl 5255–265. doi: 10.1111/j.1743-6109.2010.02012.x
[8]
Kalichman SC, Simbayi LC, Cain D, Jooste S (2009) Heterosexual anal intercourse among community and clinical settings in Cape Town, South Africa. Sex Transm Infect 85: 411–415. doi: 10.1136/sti.2008.035287
[9]
Halpern V, Ogunsola F, Obunge O, Wang CH, Onyejepu N, et al. (2008) Effectiveness of cellulose sulfate vaginal gel for the prevention of HIV infection: results of a Phase III trial in Nigeria. PLoS ONE 3: e3784. doi: 10.1371/journal.pone.0003784
[10]
Van Damme L, Govinden R, Mirembe FM, Guedou F, Solomon S, et al. (2008) Lack of effectiveness of cellulose sulfate gel for the prevention of vaginal HIV transmission. N Engl J Med 359: 463–472. doi: 10.1056/nejmoa0707957
[11]
Abdool Karim SS, Richardson BA, Ramjee G, Hoffman IF, Chirenje ZM, et al. (2011) Safety and effectiveness of BufferGel and 0.5% PRO2000 gel for the prevention of HIV infection in women. AIDS 25: 957–966. doi: 10.1097/qad.0b013e32834541d9
[12]
McCormack S, Ramjee G, Kamali A, Rees H, Crook AM, et al. (2010) PRO2000 vaginal gel for prevention of HIV-1 infection (Microbicides Development Programme 301): a phase 3, randomised, double-blind, parallel-group trial. Lancet 376: 1329–1337. doi: 10.1016/s0140-6736(10)61086-0
[13]
Kreiss J, Ngugi E, Holmes K, Ndinya-Achola J, Waiyaki P, et al. (1992) Efficacy of nonoxynol 9 contraceptive sponge use in preventing heterosexual acquisition of HIV in Nairobi prostitutes. JAMA 268: 477–482. doi: 10.1001/jama.268.4.477
[14]
Roddy RE, Zekeng L, Ryan KA, Tamoufe U, Weir SS, et al. (1998) A controlled trial of nonoxynol 9 film to reduce male-to-female transmission of sexually transmitted diseases. N Engl J Med 339: 504–510. doi: 10.1056/nejm199808203390803
[15]
Van Damme L, Ramjee G, Alary M, Vuylsteke B, Chandeying V, et al. (2002) Effectiveness of COL-1492, a nonoxynol-9 vaginal gel, on HIV-1 transmission in female sex workers: a randomised controlled trial. Lancet 360: 971–977. doi: 10.1016/s0140-6736(02)11079-8
[16]
Abdool Karim Q, Abdool Karim SS, Frohlich JA, Grobler AC, Baxter C, et al. (2010) Effectiveness and safety of tenofovir gel, an antiretroviral microbicide, for the prevention of HIV infection in women. Science 329: 1168–1174. doi: 10.1126/science.1193748
[17]
Marrazzo JM, Ramjee G, Nair G, Palanee T, Mkhize B, et al.. (2013) Pre-exposure Prophylaxis for HIV in Women: Daily Oral Tenofovir, Oral Tenofovir/Emtricitabine, or Vaginal Tenofovir Gel in the VOICE Study (MTN 003). 20th Conference on Retroviruses and Oppotunistic Infections. Atlanta, GA. #26LB.
[18]
Mayer KH, Maslankowski LA, Gai F, El-Sadr WM, Justman J, et al. (2006) Safety and tolerability of tenofovir vaginal gel in abstinent and sexually active HIV-infected and uninfected women. AIDS 20: 543–551. doi: 10.1097/01.aids.0000210608.70762.c3
[19]
Schwartz JL, Kovalevsky G, Lai JJ, Ballagh SA, McCormick T, et al. (2008) A randomized six-day safety study of an antiretroviral microbicide candidate UC781, a non-nucleoside reverse transcriptase inhibitor. Sex Transm Dis 35: 414–419. doi: 10.1097/olq.0b013e318162c4d8
[20]
Anton PA, Cranston RD, Kashuba A, Hendrix CW, Bumpus NN, et al. (2012) RMP-02/MTN-006: A phase 1 rectal safety, acceptability, pharmacokinetic, and pharmacodynamic study of tenofovir 1% gel compared with oral tenofovir disoproxil fumarate. AIDS Res Hum Retroviruses 28: 1412–1421. doi: 10.1089/aid.2012.0262
[21]
Anton PA, Saunders T, Elliott J, Khanukhova E, Dennis R, et al. (2011) First Phase 1 Double-Blind, Placebo-Controlled, Randomized Rectal Microbicide Trial Using UC781 Gel with a Novel Index of Ex Vivo Efficacy. PLoS ONE 6: e23243. doi: 10.1371/journal.pone.0023243
[22]
Rohan LC, Moncla BJ, Kunjara Na Ayudhya RP, Cost M, Huang Y, et al. (2010) In vitro and ex vivo testing of tenofovir shows it is effective as an HIV-1 microbicide. PLoS One 5: e9310. doi: 10.1371/journal.pone.0009310
[23]
Wang L, Schnaare RL, Dezzutti C, Anton PA, Rohan LC (2011) Rectal microbicides: clinically relevant approach to the design of rectal specific placebo formulations. AIDS Res Ther 8: 12. doi: 10.1186/1742-6405-8-12
[24]
Abner SR, Guenthner PC, Guarner J, Hancock KA, Cummins JE Jr, et al. (2005) A Human Colorectal Explant Culture to Evaluate Topical Microbicides for the Prevention of HIV Infection. J Infect Dis 192: 1545–1556. doi: 10.1086/462424
[25]
Fletcher PS, Elliott J, Grivel JC, Margolis L, Anton P, et al. (2006) Ex vivo culture of human colorectal tissue for the evaluation of candidate microbicides. AIDS 20: 1237–1245. doi: 10.1097/01.aids.0000232230.96134.80
[26]
Herrera C, Cranage M, McGowan I, Anton P, Shattock RJ (2009) Reverse transcriptase inhibitors as potential colorectal microbicides. Antimicrob Agents Chemother 53: 1797–1807. doi: 10.1128/aac.01096-08
[27]
Reed LJ, Muench H (1938) A simple method of estimating fifty per cent endpoints. Am J Hygiene 27: 493–497.
[28]
Koboziev I, Karlsson F, Grisham MB (2010) Gut-associated lymphoid tissue, T cell trafficking, and chronic intestinal inflammation. Ann N Y Acad Sci 1207 Suppl 1E86–93. doi: 10.1111/j.1749-6632.2010.05711.x
[29]
Langman JM, Rowland R (1986) The number and distribution of lymphoid follicles in the human large intestine. J Anat 149: 189–194.
[30]
Langman JM, Rowland R (1992) Density of lymphoid follicles in the rectum and at the anorectal junction. J Clin Gastroenterol 14: 81–84. doi: 10.1097/00004836-199201000-00020
[31]
Haase AT (2010) Targeting early infection to prevent HIV-1 mucosal transmission. Nature 464: 217–223. doi: 10.1038/nature08757
[32]
Koyanagi Y, Miles S, Mitsuyasu RT, Merrill JE, Vinters HV, et al. (1987) Dual infection of the central nervous system by AIDS viruses with distinct cellular tropisms. Science 236: 819–822. doi: 10.1126/science.3646751
[33]
Evans DF, Pye G, Bramley R, Clark AG, Dyson TJ, et al. (1988) Measurement of gastrointestinal pH profiles in normal ambulant human subjects. Gut 29: 1035–1041. doi: 10.1136/gut.29.8.1035
[34]
Buckheit RW Jr, Buckheit KW (2012) An algorithm for the preclinical development of anti-HIV topical microbicides. Curr HIV Res 10: 97–104. doi: 10.2174/157016212799304698
[35]
Lackman-Smith CS, Snyder BA, Marotte KM, Osterling MC, Mankowski MK, et al. (2010) Safety and anti-HIV assessments of natural vaginal cleansing products in an established topical microbicides in vitro testing algorithm. AIDS Res Ther 7: 22. doi: 10.1186/1742-6405-7-22
[36]
Gartner S, Markovits P, Markovitz DM, Kaplan MH, Gallo RC, et al. (1986) The role of mononuclear phagocytes in HTLV-III/LAV infection. Science 233: 215–219. doi: 10.1126/science.3014648
[37]
Grivel JC, Elliott J, Lisco A, Biancotto A, Condack C, et al. (2007) HIV-1 pathogenesis differs in rectosigmoid and tonsillar tissues infected ex vivo with CCR5- and CXCR4-tropic HIV-1. AIDS 21: 1263–1272. doi: 10.1097/qad.0b013e3281864667
[38]
Miller CJ, Li Q, Abel K, Kim EY, Ma ZM, et al. (2005) Propagation and dissemination of infection after vaginal transmission of simian immunodeficiency virus. J Virol 79: 9217–9227. doi: 10.1128/jvi.79.14.9217-9227.2005
[39]
Zhang Z, Schuler T, Zupancic M, Wietgrefe S, Staskus KA, et al. (1999) Sexual transmission and propagation of SIV and HIV in resting and activated CD4+ T cells. Science 286: 1353–1357. doi: 10.1126/science.286.5443.1353
[40]
Balandya E, Sheth S, Sanders K, Wieland-Alter W, Lahey T (2010) Semen protects CD4+ target cells from HIV infection but promotes the preferential transmission of R5 tropic HIV. J Immunol 185: 7596–7604. doi: 10.4049/jimmunol.1002846
[41]
Denison FC, Grant VE, Calder AA, Kelly RW (1999) Seminal plasma components stimulate interleukin-8 and interleukin-10 release. Mol Hum Reprod 5: 220–226. doi: 10.1093/molehr/5.3.220
[42]
Martellini JA, Cole AL, Venkataraman N, Quinn GA, Svoboda P, et al. (2009) Cationic polypeptides contribute to the anti-HIV-1 activity of human seminal plasma. FASEB J 23: 3609–3618. doi: 10.1096/fj.09-131961
[43]
Munch J, Rucker E, Standker L, Adermann K, Goffinet C, et al. (2007) Semen-derived amyloid fibrils drastically enhance HIV infection. Cell 131: 1059–1071. doi: 10.1016/j.cell.2007.10.014
[44]
Politch JA, Tucker L, Bowman FP, Anderson DJ (2007) Concentrations and significance of cytokines and other immunologic factors in semen of healthy fertile men. Hum Reprod 22: 2928–2935. doi: 10.1093/humrep/dem281
[45]
Roan NR, Munch J, Arhel N, Mothes W, Neidleman J, et al. (2009) The cationic properties of SEVI underlie its ability to enhance human immunodeficiency virus infection. J Virol 83: 73–80. doi: 10.1128/jvi.01366-08
[46]
Sabatte J, Ceballos A, Raiden S, Vermeulen M, Nahmod K, et al. (2007) Human seminal plasma abrogates the capture and transmission of human immunodeficiency virus type 1 to CD4+ T cells mediated by DC-SIGN. J Virol 81: 13723–13734. doi: 10.1128/jvi.01079-07
[47]
Introini A, Vanpouille C, Lisco A, Grivel JC, Margolis L (2013) Interleukin-7 facilitates HIV-1 transmission to cervico-vaginal tissue ex vivo. PLoS Pathog 9: e1003148. doi: 10.1371/journal.ppat.1003148
[48]
Stax MJ, van Montfort T, Sprenger RR, Melchers M, Sanders RW, et al. (2009) Mucin 6 in seminal plasma binds DC-SIGN and potently blocks dendritic cell mediated transfer of HIV-1 to CD4(+) T-lymphocytes. Virology 391: 203–211. doi: 10.1016/j.virol.2009.06.011
[49]
O’Connor TJ, Kinchington D, Kangro HO, Jeffries DJ (1995) The activity of candidate virucidal agents, low pH and genital secretions against HIV-1 in vitro. Int J STD AIDS 6: 267–272. doi: 10.1097/00002030-199411004-00196
[50]
Wolters-Everhardt E, Dony JM, Lemmens WA, Doesburg WH, De Pont JJ (1986) Buffering capacity of human semen. Fertil Steril 46: 114–119.
[51]
Mackall CL, Fry TJ, Gress RE (2011) Harnessing the biology of IL-7 for therapeutic application. Nat Rev Immunol 11: 330–342. doi: 10.1038/nri2970
[52]
Fuchs EJ, Lee LA, Torbenson MS, Parsons TL, Bakshi RP, et al. (2007) Hyperosmolar sexual lubricant causes epithelial damage in the distal colon: potential implication for HIV transmission. J Infect Dis 195: 703–710. doi: 10.1086/511279
[53]
Dezzutti CS, Brown ER, Moncla B, Russo J, Cost M, et al. (2012) Is wetter better? An evaluation of over-the-counter personal lubricants for safety and anti-HIV-1 activity. PLoS ONE 7: e48328. doi: 10.1371/journal.pone.0048328
[54]
Dezzutti CS, Hladik F (2013) Use of human mucosal tissue to study HIV-1 pathogenesis and evaluate HIV-1 prevention modalities. Curr HIV/AIDS Rep 10: 12–20. doi: 10.1007/s11904-012-0148-2
[55]
Anderson DJ, Pudney J, Schust DJ (2010) Caveats associated with the use of human cervical tissue for HIV and microbicide research. AIDS 24: 1–4. doi: 10.1097/qad.0b013e328333acfb
