Contraceiption by Targeting Germ Cell Adhesion

在整个精子发生过程中，处于不同发育阶段的发育中的生殖细胞必须附着在 生精上皮通过支持细胞-生殖细胞界面的特化细胞连接。因此，在本发明中， 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丢失， 不孕在过去的赠款期间完成的研究表明，Adjudin <$[以前称为AF- 2364，1-（2，4-二氯苄基）-7H-吲唑-3-碳酰肼]是一种很有前途的男性避孕药 因为它有效地消耗生殖细胞，特别是伸长/伸长精子细胞、圆形精子细胞， 在成年大鼠的上皮细胞中有精母细胞，但没有精原细胞。更重要的是， 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 遗传毒性研究表明，该药物的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界限很窄。到 为了避免这个问题，Adjudin与FSH突变体结合，其中内源性激素活性 突变体被剥离而不损害其FSH受体结合活性。最重要的是，其 疗效显著提高。私家侦探现在已经提出研究开发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，如凝胶贴剂或鼻喷雾剂，用于吸收，而不是父母 利用在实地和人口理事会建立的技术，一旦疗效 和生物利用度，其安全性问题将由亚慢性 在大鼠和犬中进行毒性研究，以评估其安全性和有效性之间的界限。此外，委员会认为， 生物化学，分子生物学和细胞生物学的当代技术将被用来继续 该实验室正在进行的研究，以探索Adjudin的分子作用机制，包括其 对生精上皮中支持细胞和生殖细胞的细胞效应。我们还将识别 睾丸中Adjudin的靶点，包括定位整合素中的磷酸化位点，因为其活化 很可能引发了睾丸中由Adjudin诱导的生殖细胞丢失简而言之，这一建议将继续 生产性研究在P.L.的实验室，这将导致1期临床研究的Adjudin。

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.