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Vaginal Microbicide Film Combinations of Two Reverse Transcriptase Inhibitors, EFdA and CSIC, for the Prevention of HIV-1 Sexual Transmission

Pharmaceutical Research volume 32, pages 2960–2972 (2015)Cite this article

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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Affiliations

  1. Magee Womens Research Institute, University of Pittsburgh, Pittsburgh, Pennsylvania, 15213, USA

    Wei Zhang, Minlu Hu, Yuan Shi, Tiantian Gong, Charlene S. Dezzutti, Bernard Moncla & Lisa C. Rohan

  2. Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, Pennsylvania, 15213, USA

    Wei Zhang, Minlu Hu, Yuan Shi, Tiantian Gong & Lisa C. Rohan

  3. Department of Obstetrics, Gynecology, and Reproductive Sciences, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, 15213, USA

    Charlene S. Dezzutti, Bernard Moncla & Lisa C. Rohan

  4. Department of Molecular Microbiology & Immunology, School of Medicine, University of Missouri, Columbia, Missouri, 65211, USA

    Stefan G. Sarafianos

  5. Department of Microbiology and Molecular Genetics, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, 15219, USA

    Michael A. Parniak

Authors
  1. Wei Zhang
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  2. Minlu Hu
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  3. Yuan Shi
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  4. Tiantian Gong
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  5. Charlene S. Dezzutti
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  6. Bernard Moncla
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  7. Stefan G. Sarafianos
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  8. Michael A. Parniak
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  9. Lisa C. Rohan
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Corresponding author

Correspondence to Lisa C. Rohan.

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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

KEY WORDS

  • CSIC
  • EFdA
  • HIV prevention
  • microbicide
  • vaginal film