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: 10018041
• 负责人: JAMES K CHEN
• 金额: $ 41.76万
• 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 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10018041
• 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; base; birth control; cell motility; condoms; contraceptive target; design; experimental study; homeodomain; in silico; in vivo; inhibitor/antagonist; 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; unintended pregnancy; 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; base; birth control; cell motility; condoms; contraceptive target; design; experimental study; homeodomain; in silico; in vivo; inhibitor/antagonist; 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; 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).

必须采用安全，有效和可逆的避孕方法来解决8500万 每年全球发生的计划外怀孕。除了这些意外的负面影响 关于全球可持续性的怀孕，这些案件中几乎五分之一是通过不安全的堕胎终止的 妇女健康的风险很大。自1950年代发明“药丸”以来，绝大多数出生 控制选项是由女性指导的，包括雌激素或孕激素治疗，屏障方法， 插入设备和输卵管连接。相比之下，男性仍然仅限于避孕套和输精管切除术 高失败率和不完全可逆性。 男性避孕的药理学策略将有助于与当前的女性指导达到平等 选项。但是，基于马的疗法会导致代谢性疾病，情绪变化，血栓形成，痤疮， 和有证明的变性。发育中的非激素剂，例如视黄酸信号拮抗剂， 孤田胺衍生物和溴结构域特异性蛋白抑制剂也可能具有不良目标 副作用。确定精子开发和功能的新调节剂对于弥合这一差距， 和可吸毒的睾丸特异性蛋白质是特别有吸引力的靶标。我们的项目专注于一个信号 类似于此范式的蛋白质：Hipk4，同源域相互作用蛋白激酶家族的成员 这是在开发精子中表达的。我们观察到缺乏HIPK4功能的雄性小鼠是不育的，但 否则似乎具有正常的发展，生理和行为。 HIPK4缺陷小鼠展示 与少肠刺激性缺陷相一致的精子缺陷，其Missshapeen精子是 体外受精无能力。我们的调查进一步表明HIPK4调节肌动蛋白驱动的头部 精子伸长期间的塑形。 我们的发现强调了小分子HIPK4抑制剂作为非激素雄性的潜力 避孕药具，尤其是针对已配置的ATP结合口袋之外的拮抗剂。 为了实现这一目标，我们将开发变构HIPK4抑制剂，并评估它们对精子发生的影响 在动物模型中的男性生育能力。该项目的R61阶段将着重于建立一个工作流程 识别和表征变构HIPK4拮抗剂，包括主要的蛋白质热移（PTS） 测定（R61 AIM 1）和抑制剂效力和选择性的二级/第三次测定（R61 AIM 2）。我们也会 在计算机和晶体学方案中开发，用于研究HIPK4抑制的结构基础（R61 AIM 3）。 完成这些里程碑后，我们将追求该项目的R33阶段，其中包括一个大规模的， 变构HIPK4抑制剂（R33 AIM 1）的高通量PTS屏幕，并通过 药物化学和基于结构的设计（R33 AIM 2）。然后，我们将评估HIPK4拮抗剂 动物模型以确定其作为男性避孕药的安全性，效率和可逆性（R33 AIM 3）。