title: Two new male contraceptives exert their effects by depleting germ cells prematurely from the testis.
abstract: The three currently available male contraceptive approaches are 1) the barrier method such as the condom, 2) hormonal methods by disrupting the pituitary-testicular axis so as to impair spermatogenesis, and 3) immunological methods by preparing vaccines against male-specific antigens. We hereby describe an alternative approach in which attachments of developing germ cells onto the seminiferous epithelium are disrupted, thereby inducing their premature release into the tubular lumen. This in turn leads to infertility. A panel of analogues based on the core structure of 1-(2,4-dichlorobenzyl)-indazole-3-carboxylic acid was synthesized. These compounds were subjected to an in vivo screening assay assessing their effects in inducing the expression of testin, a testicular marker whose expression correlates with the integrity of Sertoli-germ cell junctions. An induction of testin expression in the testis signifies a disruption of Sertoli-germ cell junctions that is followed by depletion of germ cells from the seminiferous epithelium. Two compounds, namely 1-(2,4-dichlorobenzyl)-indazole-3-carbohydrazide (AF-2364) and 1-(2,4-dichlorobenzyl)-indazole-3-acrylic acid (AF-2785), were identified that caused detachment of germ cells, in particular round and elongated spermatids, from the epithelium inducing their premature release into the tubular lumen as confirmed by histological analysis. Adult rats receiving several oral doses of either one of these compounds became infertile within 3-7 wk after the epididymal sperm reserve was exhausted. Depending on the dosing of the administered compound, rats became infertile for 4-14 wk before their fertility gradually bounced back, illustrating the reversibility and efficacy of these new compounds. Also, these compounds did not appear to impair the hypothalamus-pituitary-testicular axis because the serum levels of LH, FSH, and testosterone of the treated animals did not change significantly when compared to control rats. In addition, results of serum microchemistry illustrate that liver and kidney function was not affected in animals treated with both compounds.
authors.affiliation: Population Council, The Rockefeller University, New York, New York 10021, USA. yan@popcbr.rockefeller.edu
authors.forename: C Y
authors.initials: CY
authors.lastname: Cheng
authors.forename: B
authors.initials: B
authors.lastname: Silvestrini
authors.forename: J
authors.initials: J
authors.lastname: Grima
authors.forename: M Y
authors.initials: MY
authors.lastname: Mo
authors.forename: L J
authors.initials: LJ
authors.lastname: Zhu
authors.forename: E
authors.initials: E
authors.lastname: Johansson
authors.forename: L
authors.initials: L
authors.lastname: Saso
authors.forename: M G
authors.initials: MG
authors.lastname: Leone
authors.forename: M
authors.initials: M
authors.lastname: Palmery
authors.forename: D
authors.initials: D
authors.lastname: Mruk
mesh_terms: D000818:Animals; D001593:Benzyl Compounds / Q000008:administration & dosage / Q000032:analysis / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D002452:Cell Count; D003272:Contraceptive Agents, Male / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005640:Follicle Stimulating Hormone / Q000097:blood; D015870:Gene Expression / Q000187:drug effects; D006834:Hydrazines / Q000008:administration & dosage / Q000032:analysis / Q000494:pharmacology; D007150:Immunohistochemistry; D007191:Indazoles / Q000008:administration & dosage / Q000032:analysis / Q000494:pharmacology; D007668:Kidney / Q000187:drug effects; D008099:Liver / Q000187:drug effects; D007986:Luteinizing Hormone / Q000097:blood; D008297:Male; D009929:Organ Size / Q000187:drug effects; D011506:Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D012670:Seminiferous Epithelium / Q000166:cytology; D013087:Spermatids / Q000187:drug effects / Q000502:physiology; D013094:Spermatozoa / Q000187:drug effects* / Q000502:physiology; D013737:Testis / Q000166:cytology*; D013739:Testosterone / Q000097:blood
chemical_list: C415442:1-(2,4-dichlorobenzyl)indazole-3-acrylic acid; C432482:1-(2,4-dichlorobenzyl)indazole-3-carbohydrazide; D001593:Benzyl Compounds; D003272:Contraceptive Agents, Male; D006834:Hydrazines; D007191:Indazoles; D011506:Proteins; D012333:RNA, Messenger; C489180:testin protein, rat; D013739:Testosterone; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone