Development of allosteric HIPK4 inhibitors as non-hormonal male contraceptives

开发变构 HIPK4 抑制剂作为非激素男性避孕药

• 批准号: 10470960
• 负责人: JAMES K CHEN
• 金额: $ 87.06万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-15 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10470960
• 关键词: Acne; Actins; Address; Algorithms; Allosteric Site; Animal Model; Atrophic; Barrier Contraception; Behavior; Binding; Biochemical; Biological; Biological Assay; Biological Availability; Biology; Blood-Testis Barrier; Bromodomain; Catalogs; Cells; Characteristics; Chemicals; Clinical; Contraceptive methods; Crystallization; Crystallography; Defect; Development; Docking; Drug Kinetics; Enzymes; Estrogens; Event; Exhibits; Failure; Family; Family member; Female; Fertility Disorders; Fertilization in Vitro; Genetic; Goals; Head; Homology Modeling; Hormonal; Hormones; In Vitro; Infertility; Intrauterine Devices; Investigation; Knockout Mice; Lead; Libraries; Lonidamine; Male Contraceptive Agents; Metabolic; Metabolic Diseases; Metabolism; Methods; Moods; Mus; Oocytes; Oral; Penetrance; Pharmaceutical Chemistry; Pharmacology; Phase; Phosphotransferases; Physiology; Pilot Projects; Progestins; Protein Kinase; Proteins; Protocols documentation; Recombinants; Reproductive Biology; Reproductive Health; Resources; Risk; Rodent Model; Safety; Shapes; Signal Transduction; Signaling Protein; Spermatids; Spermiogenesis; Structure; Testing; Testis; Thrombosis; Toxic effect; Tretinoin; Tubal Ligation; Variant; Vasectomy; Women' s Health; abortion; absorption; affective disturbance; analog; antagonist; base; birth control; cell motility; condoms; contraceptive target; design; experimental study; homeodomain; in silico; in vivo; inhibitor; invention; kinase inhibitor; lead optimization; male; male fertility; member; men; mutant; novel; parity; pill; prevent; scaffold; screening; side effect; small molecule; sperm cell; success; synchrotron radiation; therapeutic development; therapeutic lead compound; thrombotic; unintended pregnancy

Safe, effective, and reversible methods for contraception are necessary to address the 85 million unplanned pregnancies that occur worldwide each year. In addition to the negative impact of these unintended pregnancies on global sustainability, nearly one-fifth of these cases are terminated through unsafe abortions with significant risks to women’s health. Since the invention of “the Pill” in the 1950s, the vast majority of birth control options have been female-directed, including estrogen or progestin treatments, barrier methods, intrauterine devices, and tubal ligation. In contrast, men remain limited to condoms and vasectomy, which have high failure rates and incomplete reversibility, respectively. Pharmacological strategies for male contraception would help achieve parity with current female-directed options. However, hormone-based therapies can lead to metabolic disorders, mood changes, thrombosis, acne, and testicular degeneration. Non-hormonal agents in development such as retinoic acid signaling antagonists, lonidamine derivatives, and bromodomain testis-specific protein inhibitors can also have undesirable on-target side effects. Identifying new regulators of sperm development and function will be necessary to bridge this gap, and druggable testis-specific proteins are especially attractive targets. Our project focuses on one signaling protein that exemplifies this paradigm: HIPK4, a member of the homeodomain-interacting protein kinase family that is expressed in developing sperm. We observe that male mice lacking HIPK4 function are infertile but otherwise appear to have normal development, physiology, and behavior. HIPK4-deficient mice exhibit spermatogenic defects that are consistent with oligoasthenoteratozoospermia, and their misshapen sperm are incompetent for in vitro fertilization. Our investigations further indicate that HIPK4 regulates actin-driven head shaping during spermatid elongation. Our findings underscore the potential of small-molecule HIPK4 inhibitors as non-hormonal male contraceptives, particularly antagonists that target regions outside of the conserved ATP-binding pocket. Toward this goal, we will develop allosteric HIPK4 inhibitors and evaluate their effects on spermiogenesis and male fertility in animal models. The R61 phase of this project will focus on establishing a workflow for identifying and characterizing allosteric HIPK4 antagonists, including a primary protein thermal shift (PTS) assay (R61 Aim 1) and secondary/tertiary assays of inhibitor potency and selectivity (R61 Aim 2). We will also develop in silico and crystallographic protocols for studying the structural basis of HIPK4 inhibition (R61 Aim 3). After completing these milestones, we will pursue the R33 phase of this project, which includes a large-scale, high-throughput PTS screen for allosteric HIPK4 inhibitors (R33 Aim 1) and hit-to-lead optimization through medicinal chemistry and structure-based design (R33 Aim 2). We will then evaluate HIPK4 antagonists in animal models to determine their safety, efficacy, and reversibility as male contraceptives (R33 Aim 3).

安全、有效和可逆的避孕方法是解决8500万人口问题的必要条件。 每年全球范围内都会发生意外怀孕。除了这些非预期的负面影响之外， 在全球可持续性的怀孕中，近五分之一的病例是通过不安全堕胎终止的。 对女性健康有重大风险。自20世纪50年代发明“避孕药”以来， 控制选择是针对女性的，包括雌激素或孕酮治疗，屏障方法， 子宫内避孕器和输卵管结扎相比之下，男性仍然仅限于使用避孕套和输精管结扎术， 高故障率和不完全可逆性。 男性避孕的药理学策略将有助于实现与目前女性指导的 选项.然而，基于维生素E的疗法可能导致代谢紊乱、情绪变化、血栓形成、痤疮， 和睾丸退化正在开发的非激素类药物，如视黄酸信号拮抗剂， 氯尼达明衍生物和布罗莫结构域睾丸特异性蛋白抑制剂也可具有不期望的靶向作用。 副作用.确定精子发育和功能的新调节因子对于弥合这一差距是必要的， 可药物化的睾丸特异性蛋白是特别有吸引力的靶点。我们的项目集中在一个信号 一种证实这种模式的蛋白质：HIPK 4，同源结构域相互作用蛋白激酶家族的成员 在发育中的精子中表达。我们观察到缺乏HIPK 4功能的雄性小鼠是不育的， 在其他方面表现出正常的发育、生理和行为。HIPK 4缺陷小鼠表现出 生精缺陷符合少弱畸形精子症，他们畸形的精子是 不能进行体外受精我们的研究进一步表明，HIPK 4调节肌动蛋白驱动的头部 在精子细胞伸长过程中形成。 我们的研究结果强调了小分子HIPK 4抑制剂作为非激素男性的潜力 避孕药，特别是靶向保守ATP结合口袋以外区域的拮抗剂。 为了实现这一目标，我们将开发别构HIPK 4抑制剂，并评估其对精子发生的影响， 动物模型中的雄性生育力。该项目的R61阶段将侧重于建立一个工作流程， 鉴定和表征别构HIPK 4拮抗剂，包括初级蛋白质热位移（PTS） 抑制剂效价和选择性的二级/三级测定（R61 Aim 1）。我们还将 开发用于研究HIPK 4抑制的结构基础的计算机和晶体学方案（R61 Aim 3）。 在完成这些里程碑之后，我们将继续进行该项目的R33阶段，其中包括一个大规模的， 用于别构HIPK 4抑制剂（R33 Aim 1）高通量PTS筛选和通过以下方法的命中-先导物优化 药物化学和基于结构的设计（R33 Aim 2）。然后，我们将评估HIPK 4拮抗剂， 动物模型，以确定其作为男性避孕药的安全性、有效性和可逆性（R33目标3）。