Candidate Topical Microbicides Bind Herpes Simplex Virus Glycoprotein B and Prevent Viral Entry and Cell-to-Cell Spread

ABSTRACT Topical microbicides designed to prevent acquisition of sexually transmitted infections are urgently needed. Nonoxynol-9, the only commercially available spermicide, damages epithelium and may enhance human immunodeficiency virus transmission. The observation that herpes simplex virus (HSV) and human immunodeficiency virus bind heparan sulfate provided the rationale for the development of sulfated or sulfonated polymers as topical agents. Although several of the polymers have advanced to clinical trials, the spectrum and mechanism of anti-HSV activity and the effects on soluble mediators of inflammation have not been evaluated. The present studies address these gaps. The results indicate that PRO 2000, polystyrene sulfonate, cellulose sulfate, and polymethylenehydroquinone sulfonate inhibit HSV infection 10,000-fold and are active against clinical isolates, including an acyclovir-resistant variant. The compounds formed stable complexes with glycoprotein B and inhibit viral binding, entry, and cell-to-cell spread. The effects may be long lasting due to the high affinity and stability of the sulfated compound-virus complex, as evidenced by surface plasmon resonance studies. The candidate microbicides retained their antiviral activities in the presence of cervical secretions and over a broad pH range. There was little reduction in cell viability following repeated exposure of human endocervical cells to these compounds, although a reduction in secretory leukocyte protease inhibitor levels was observed. These studies support further development and rigorous evaluation of these candidate microbicides.

[1]  P. Harrison,et al.  Topical microbicides for disease prevention: status and challenges. , 2003, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[2]  R. Puri,et al.  Human Cytokines: Their Role in Disease and Therapy , 1995 .

[3]  B. Herold,et al.  Development of Topical Microbicides for Prevention of Human Immunodeficiency Virus and Herpes Simplex Virus , 2003, American journal of reproductive immunology.

[4]  John D Lambris,et al.  Kinetic analysis of glycoprotein C of herpes simplex virus types 1 and 2 binding to heparin, heparan sulfate, and complement component C3b. , 2002, Virology.

[5]  R. Hayes,et al.  Interactions between herpes simplex virus type 2 and human immunodeficiency virus type 1 infection in African women: opportunities for intervention. , 2000, The Journal of infectious diseases.

[6]  L. Zaneveld,et al.  Study of the vaginal tolerance to Acidform, an acid-buffering, bioadhesive gel. , 1999, Contraception.

[7]  H. Rees,et al.  Safety and tolerability of vaginal PRO 2000 gel in sexually active HIV-uninfected and abstinent HIV-infected women , 2003, AIDS.

[8]  Myron S. Cohen,et al.  Sexually transmitted diseases enhance HIV transmission: no longer a hypothesis , 1998, The Lancet.

[9]  R. Lehrer,et al.  NP-1, a rabbit alpha-defensin, prevents the entry and intercellular spread of herpes simplex virus type 2. , 2003, Antimicrobial agents and chemotherapy.

[10]  A. Wald,et al.  Herpes simplex virus: the importance of asymptomatic shedding. , 2000, The Journal of antimicrobial chemotherapy.

[11]  D. Marcellino,et al.  Poly(sodium 4-styrene sulfonate): an effective candidate topical antimicrobial for the prevention of sexually transmitted diseases. , 2000, The Journal of infectious diseases.

[12]  H. Rees,et al.  Effectiveness of COL-1492, a nonoxynol-9 vaginal gel, on HIV-1 transmission in female sex workers: a randomised controlled trial , 2002, The Lancet.

[13]  C. Crumpacker,et al.  Characterization of an acyclovir-resistant herpes simplex virus type 2 strain isolated from a premature neonate. , 2000, The Journal of infectious diseases.

[14]  S. S. Olmsted,et al.  Tests of BufferGel for Contraception and Prevention of Sexually Transmitted Diseases in Animal Models , 2001, Sexually transmitted diseases.

[15]  Robert A. Anderson,et al.  Properties of a new acid-buffering bioadhesive vaginal formulation (ACIDFORM). , 2001, Contraception.

[16]  J. H. van de Wijgert,et al.  Phase 1 Trial of the Topical Microbicide BufferGel: Safety Results From Four International Sites , 2001, Journal of acquired immune deficiency syndromes.

[17]  D. Marcellino,et al.  Mandelic Acid Condensation Polymer: Novel Candidate Microbicide for Prevention of Human Immunodeficiency Virus and Herpes Simplex Virus Entry , 2002, Journal of Virology.

[18]  S. Wahl,et al.  Inhibition of human immunodeficiency virus type 1 infectivity by secretory leukocyte protease inhibitor occurs prior to viral reverse transcription. , 1997, Blood.

[19]  R. Longnecker,et al.  Herpesvirus Entry: an Update , 2003, Journal of Virology.

[20]  S. Straus,et al.  Specificity and affinity of binding of herpes simplex virus type 2 glycoprotein B to glycosaminoglycans , 1997, Journal of virology.

[21]  R. Roller,et al.  Characterization of a BHK(TK-) cell clone resistant to postattachment entry by herpes simplex virus types 1 and 2 , 1997, Journal of virology.

[22]  D. Anderson,et al.  The molecular basis of nonoxynol-9-induced vaginal inflammation and its possible relevance to human immunodeficiency virus type 1 transmission. , 2001, The Journal of infectious diseases.

[23]  D. Marcellino,et al.  Bile Salts: Natural Detergents for the Prevention of Sexually Transmitted Diseases , 1999, Antimicrobial Agents and Chemotherapy.

[24]  L. van Damme,et al.  Safety of multiple daily applications of COL-1492, a nonoxynol-9 vaginal gel, among female sex workers , 2000, AIDS.

[25]  J. Turpin Considerations and development of topical microbicides to inhibit the sexual transmission of HIV , 2002, Expert opinion on investigational drugs.

[26]  B. Herold,et al.  Glycoprotein C of herpes simplex virus type 1 plays a principal role in the adsorption of virus to cells and in infectivity , 1991, Journal of virology.

[27]  R. Eisenberg,et al.  Examination of the Kinetics of Herpes Simplex Virus Glycoprotein D Binding to the Herpesvirus Entry Mediator, Using Surface Plasmon Resonance , 1998, Journal of Virology.

[28]  D. Anderson,et al.  Differential expression of immunobiological mediators by immortalized human cervical and vaginal epithelial cells. , 1999, Biology of reproduction.

[29]  L. van Damme,et al.  Safety and Acceptability of Penile Application of 2 Candidate Topical Microbicides: BufferGel and PRO 2000 Gel: 3 Randomized Trials in Healthy Low‐Risk Men and HIV‐Positive Men , 2003, Journal of acquired immune deficiency syndromes.

[30]  M. Hirsch,et al.  Naphthalene sulfonate polymers with CD4-blocking and anti-human immunodeficiency virus type 1 activities , 1996, Antimicrobial agents and chemotherapy.

[31]  R. Sekulovich,et al.  Limited variability of glycoprotein gene sequences and neutralizing targets in herpes simplex virus type 2 isolates and stability on passage in cell culture. , 1998, The Journal of infectious diseases.

[32]  D. Bernstein,et al.  The topical microbicide PRO 2000 protects against genital herpes infection in a mouse model. , 1999, The Journal of infectious diseases.

[33]  F. Krebs,et al.  A Broad-Spectrum Microbicide with Virucidal Activity against Sexually Transmitted Viruses , 1999, Antimicrobial Agents and Chemotherapy.

[34]  B. Cullen,et al.  Molecular basis of latency in pathogenic human viruses. , 1991, Science.

[35]  Robert A. Anderson,et al.  Preclinical evaluation of sodium cellulose sulfate (Ushercell) as a contraceptive antimicrobial agent. , 2002, Journal of andrology.

[36]  G. Doncel,et al.  Evaluation of poly(styrene-4-sulfonate) as a preventive agent for conception and sexually transmitted diseases. , 2000, Journal of andrology.

[37]  P. Spear,et al.  Initial interaction of herpes simplex virus with cells is binding to heparan sulfate , 1989, Journal of virology.

[38]  D. Malamud,et al.  Sodium Dodecyl Sulfate and C31G as Microbicidal Alternatives to Nonoxynol 9: Comparative Sensitivity of Primary Human Vaginal Keratinocytes , 2000, Antimicrobial Agents and Chemotherapy.

[39]  G. Wilbanks,et al.  Sulfated carbohydrate compounds prevent microbial adherence by sexually transmitted disease pathogens , 1997, Antimicrobial agents and chemotherapy.

[40]  D. Phillips,et al.  Sulfated polysaccharides inhibit lymphocyte-to-epithelial transmission of human immunodeficiency virus-1. , 1996, Biology of reproduction.

[41]  N. Cheshenko,et al.  Glycoprotein B plays a predominant role in mediating herpes simplex virus type 2 attachment and is required for entry and cell-to-cell spread. , 2002, The Journal of general virology.