Contraceiption by Targeting Germ Cell Adhesion

通过靶向生殖细胞粘附来避孕

• 批准号: 7284737
• 负责人: C. Yan Cheng
• 金额: $ 37.98万
• 依托单位: POPULATION COUNCIL
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-03-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7284737
• 关键词: AF2364; Acute; Adult; Aldehydes; Amino Acids; Apoptosis; Biochemistry; Biological Availability; Canis familiaris; Carboxylic Acids; Cell Adhesion; Cells; Cellular biology; Clinical Research; Collaborations; Contraceptive Agents; Contraceptive methods; Development; Disruption; Dose; Drug Kinetics; Energy Metabolism; Epithelium; Escherichia coli; Fertility; Follicle Stimulating Hormone; Follicle Stimulating Hormone Receptor; Gel; Germ Cells; Goals; Grant; Guanosine Monophosphate; Guidelines; Hormonal; Hydrazones; Hypothalamic structure; In Vitro; Indazoles; Infertility; Integrins; Intercellular Junctions; Laboratories; Lead; Licensing; Lonidamine; Male Contraceptions; Male Contraceptive Agents; Maps; Membrane; Molecular Biology; Molecular Mechanisms of Action; Mutate; Nose; Object Attachment; Pharmacologic Substance; Phase; Phosphoric Monoester Hydrolases; Phosphorylation Site; Phosphotransferases; Pituitary Gland; Population; Production; Protein Biosynthesis; Proteins; Radioimmunoassay; Rattus; Recombinants; Research; Route; Safety; Signal Transduction; Site; Sodium; Spermatids; Spermatocytes; Spermatogenesis; Spermatogonia; Staging; System; Techniques; Technology; Testis; Time; Title; Toxic effect; United States Food and Drug Administration; Week; Work; absorption; carbohydrazide; cellular targeting; contraceptive efficacy; cost; design; genotoxicity; glycosylation; improved; in vivo; male; men; metaperiodate; mutant; receptor binding; sertoli cell; spermatogenic epithelium structure

Throughout spermatogenesis, developing germ cells at different stages of their development must attach to the seminiferous epithelium via specialized cell junctions at the Sertoli-germ cell interface. As such, disruption of germ cell adhesion, even transiently, can lead to germ cell loss from the epithelium, resulting in infertility. Studies completed during the past grant period have shown that Adjudin¿ [formerly called AF- 2364, 1-(2,4-dichlorobenzyl)-7H-indazole-3-carbohydrazide] is a promising candidate for male contraception since it effectively depletes germ cells, particularly elongating/elongate spermatids, round spermatids, and spermatocytes, but not spermatogonia, from the epithelium in adult rats. More important, studies performed by licensed toxicologists according to FDA guidelines to assess the acute toxicity, mutagenicity, and genotoxicity of Adjudin have shown that it is safe for its further development. In a subsequent subchronic toxicity study, however, it was shown that Adjudin has a narrow margin between its safety and efficacy. To circumvent this issue, Adjudin was conjugated to an FSH mutant in which the intrinsic hormonal activity of the mutant was stripped without compromising its FSH receptor binding activity. Most importantly, its efficacy was significantly improved. The P.I. has now proposed studies to develop techniques for GMP production of this Adjudin-FSH mutant conjugate in collaboration with an industrial partner, and to develop alternative administrative routes, such as a gel patch or nasal spray for its absorption instead of parental administration, using technologies established in the field and at the Population Council. Once the efficacy and bioavailability of the conjugate are established, its safety issue will be carefully evaluated by subchronic toxicity studies in rats and dogs to assess the margin between its safety and efficacy. Furthermore, contemporary techniques of biochemistry, molecular biology and cell biology will be used to continue the ongoing research in this laboratory to probe the molecular mechanism(s) of action of Adjudin including its cellular effects on Sertoli and germ cells in the seminiferous epithelium. We will also identify the cellular target(s) of Adjudin in the testis, including mapping the phosphorylation site(s) in integrin, since its activation likely triggers the Adjudin-induced germ cell loss from the testis. In short, this proposal will continue the productive research in the P.l.'s laboratory, which will lead to a Phase 1 clinical study of Adjudin.

在整个精子发生过程中，处于不同发育阶段的生殖细胞必须附着在 通过支持细胞-生殖细胞界面处的特殊细胞连接来连接生精上皮。像这样， 生殖细胞粘附的破坏，即使是短暂的，也会导致生殖细胞从上皮细胞中流失，从而导致 不孕症。在过去的资助期内完成的研究表明，Adjudin¿ [以前称为 AF- 2364, 1-(2,4-二氯苄基)-7H-吲唑-3-碳酰肼]是一种有前途的男性避孕候选药物 因为它能有效地消耗生殖细胞，特别是伸长/伸长的精子细胞、圆形精子细胞和 来自成年大鼠上皮的精母细胞，但不是精原细胞。更重要的是，进行的研究 由有执照的毒理学家根据 FDA 指南评估急性毒性、致突变性和 Adjudin 的遗传毒性表明其进一步开发是安全的。在随后的亚慢性 然而，毒性研究表明 Adjudin 的安全性和有效性之间的差距很小。到 为了规避这个问题，Adjudin 与 FSH 突变体结合，其中内在的激素活性 突变体被剥离而不影响其 FSH 受体结合活性。最重要的是，它的 疗效显着提高。 P.I.现在已提出研究开发 GMP 技术 与工业合作伙伴合作生产这种 Adjudin-FSH 突变体缀合物，并开发 替代给药途径，例如凝胶贴剂或鼻喷雾剂以代替父母的吸收 管理，利用实地和人口理事会建立的技术。一旦见效 结合物的生物利用度确定后，将通过亚慢性仔细评估其安全性问题 对大鼠和狗进行毒性研究，以评估其安全性和有效性之间的界限。此外， 将利用生物化学、分子生物学和细胞生物学的现代技术来继续 本实验室正在进行的研究旨在探讨 Adjudin 的分子作用机制，包括其作用 对生精上皮中的支持细胞和生殖细胞的细胞作用。我们还将识别蜂窝 睾丸中 Adjudin 的靶标，包括自激活以来整合素中磷酸化位点的图谱 可能会引发 Adjudin 诱导的睾丸生殖细胞损失。简而言之，该提案将继续 P.l. 实验室的富有成效的研究将导致 Adjudin 的一期临床研究。