Abstract
Purpose
EFdA is a potent nucleoside reverse transcriptase inhibitor (NRTI) with activity against a wide spectrum of wild-type and drug resistant HIV-1 variants. CSIC is a tight-binding non-nucleoside reverse transcriptase inhibitor (NNRTI) with demonstrated anti-HIV properties important for use in topical prevention of HIV transmission. The objective of this study was to develop and characterize film-formulated EFdA and CSIC for use as a female-controlled vaginal microbicide to prevent sexual transmission of HIV.
Methods
Assessments of EFdA- and CSIC-loaded films included physicochemical characteristics, in vitro cytotoxicity, epithelia integrity studies, compatibility with the normal vaginal Lactobacillus flora and anti-HIV bioactivity evaluations.
Results
No significant change in physicochemical properties or biological activity of the combination films were noted during 3 months storage. In vitro cytotoxicity and bioactivity testing showed that 50% cytotoxic concentration (CC50) of either EFdA or CSIC was several orders of magnitude higher than the 50% effective concentration (EC50) values. Film-formulated EFdA and CSIC combination showed additive inhibitory activity against wild type and drug-resistant variants of HIV. Epithelial integrity studies demonstrated that the combination vaginal film had a much lower toxicity to HEC-1A monolayers compared to that of VCF®, a commercial vaginal film product containing nonoxynol-9. Polarized ectocervical explants showed films with drug alone or in combination were effective at preventing HIV infection.
Conclusions
Our data suggest that vaginal microbicide films containing a combination of the NRTI EFdA and the NNRTI CSIC have potential to prevent HIV-1 sexual transmission.
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Abbreviations
- CSIC:
-
5-chloro-3-phenylsulfonylindole-2-carboxamide
- EFdA:
-
4′-ethynyl-2-fluoro-2′-deoxyadenosine
- FBS:
-
Fetal bovine serum
- FESEM:
-
Field emission scanning electron microscopy
- HIV:
-
Human immunodeficiency virus
- HPMC:
-
Hydroxypropyl methyl cellulose
- HSV:
-
Herpes simplex virus
- LOD:
-
Limit of detection
- LOQ:
-
Limit of quantification
- NNRTI:
-
Non-nucleoside reverse transcriptase inhibitor
- NRTI:
-
Nucleoside reverse transcriptase inhibitor
- PEG:
-
Polyethylene glycol
- PVA:
-
Polyvinyl alcohol
- RSD:
-
Relative standard deviation
- TEER:
-
Transepithelial electrical resistance
- XRD:
-
X-ray diffraction
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ACKNOWLEDGMENTS AND DISCLOSURES
We thank Ryan Winstead for his assistance with SEM study. We also would like to thank Eva Nagy for her help with bioactivity studies. Research reported in this publication was supported in part by grants AI076119 and AI079801 from the National Institute of Allergy and Infectious Diseases of the National Institutes of Health.
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Zhang, W., Hu, M., Shi, Y. et al. Vaginal Microbicide Film Combinations of Two Reverse Transcriptase Inhibitors, EFdA and CSIC, for the Prevention of HIV-1 Sexual Transmission. Pharm Res 32, 2960–2972 (2015). https://doi.org/10.1007/s11095-015-1678-2
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DOI: https://doi.org/10.1007/s11095-015-1678-2