Targeting Na,K-ATPase alpha4 for male contraception

靶向 Na,K-ATPase α4 的男性避孕

• 批准号: 10618848
• 负责人: V GUSTAVO BLANCO
• 金额: $ 53.04万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-15 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10618848
• 关键词: ATPase inhibitory protein; Affect; Affinity; Animals; Antihistamines; Appearance; Area; Binding; Binding Proteins; Biochemical; Biological; Biological Availability; Biological Markers; Cell membrane; Cells; Characteristics; Chemicals; Clinical; Clinical Research; Contraceptive Agents; Contraceptive methods; Cytoplasm; Data; Drug Kinetics; Drug Targeting; Effectiveness; Ejaculation; Epididymis; Ethers; Female; Female genitalia; Fertility; Fertilization in Vitro; Future; Gland; Goals; Human; Hyperactivity; In Vitro; Infertility; Investigational Drugs; Ions; K ATPase; Knockout Mice; Lead; Male Contraceptive Agents; Male Infertility; Maximum Tolerated Dose; Meiosis; Membrane Potentials; Metabolic; Metabolism; Mus; Mutation; Oral; Oral Administration; Oral Contraceptives; Ouabain; Partner in relationship; Permeability; Pharmaceutical Chemistry; Pilot Projects; Plasma; Population; Property; Protein Isoforms; Proteins; Public Health; Research; Safety; Series; Site; Solubility; Source; Specificity; Sperm Capacitation; Sperm Motility; Sperm Tail; Spermatocytes; Spermatogenesis; Sterility; Surface; System; Testing; Testis; Toxic effect; Validation; Woman; absorption; analog; birth control; cell motility; druggable target; efficacy study; experimental study; extracellular; improved; in silico; in vivo; inhibitor; male; male fertility; men; molecular size; monolayer; novel; pharmacologic; pre-clinical; preclinical study; progenitor; programs; reproductive tract; reversible contraceptive; scaffold; scale up; side effect; small molecule; sperm cell; sperm function; unintended pregnancy; virtual screening

PROJECT SUMMARY / ABSTRACT The rapidly growing world population and the high rate of unintended pregnancies make contraception a need and a priority for any public health program. While several contraceptive methods, with varying efficacy are currently available for women, a more comprehensive approach to birth control requires extending contraception to males. However, a safe, effective and reversible contraceptive for men is still unavailable. An attractive approach to develop male contraceptives consists in targeting proteins that are specifically expressed in sperm and are required for sperm fertility. We have shown that Na,K-ATPase α4 (NKAα4), a plasma membrane ion transporter which exchanges cytoplasmic Na+ for extracellular K+, is a validated target for male contraception. NKAα4 is uniquely expressed in testis male germ cells after meiosis, is particularly abundant in the sperm flagellum, and is critical for sperm function. Deletion of NAKα4 in mice results in complete sterility of only the male but not the female animals. NKAα4 is essential for sperm motility and sperm capacitation. Its activity maintains sperm intracellular Na+ levels ([Na+]i) and several vital sperm parameters, including membrane potential (Vm), intracellular Ca+2 ([Ca2+]i) and pH. From a biochemical standpoint, NKAα4 has a particularly high affinity for ouabain, the specific inhibitor of Na,K-ATPase. We took advantage of this property to specifically target NKAα4 and block its function to achieve male infertility. We synthesized a series of small molecule compounds, which can selectively bind to the high ouabain affinity site of NKAα4. Some of these compounds inhibit NKAα4 and affect sperm motility both in vitro and after administration to mice. This provides strong evidence for the suitability of NKAα4 as a pharmacological target and our compounds as agents that can be used for the control of male fertility. However, before NKAα4 inhibitors can be moved forward into their application as male contraceptives, it is necessary that their efficacy, drug-target interaction, biomarkers for their in vitro and vivo specificity, side effects, mechanisms of action and pharmacokinetic parameters are identified and optimized for future clinical use. We will test this in two aims. In specific aim 1, we will develop compounds with the capacity to selectively inhibit NKAα4 and block sperm function, which will be ready for testing in mice. Then, during specific aim 2, we will perform studies to obtain preclinical validation to advance the NKAα4 inhibitors as male contraceptives. A series of rigorous approaches from the medicinal chemistry and biological areas will be used to identify the compounds which will have the characteristics that will be necessary for male contraception. This research will be essential to fulfill the highly desired unmet goal of obtaining a non-hormonal pharmacological agent that could be used as an oral, reversible agent for the control of male fertility.

项目摘要/摘要

项目成果

Design, Synthesis, and in Vitro and in Vivo Evaluation of Ouabain Analogues as Potent and Selective Na,K-ATPase α4 Isoform Inhibitors for Male Contraception.

• DOI: 10.1021/acs.jmedchem.7b00925
• 发表时间: 2018-03-08
• 期刊: Journal of medicinal chemistry
• 影响因子: 7.3
• 作者: Syeda SS;Sánchez G;Hong KH;Hawkinson JE;Georg GI;Blanco G
• 通讯作者: Blanco G

Na,K-ATPase α4, and Not Na,K-ATPase α1, is the Main Contributor to Sperm Motility, But its High Ouabain Binding Affinity Site is Not Required for Male Fertility in Mice.

Na,K-ATPase α4（而不是 Na,K-ATPase α1）是精子活力的主要贡献者，但其高哇巴因结合亲和力位点并不是小鼠雄性生育能力所必需的。

• DOI: 10.1007/s00232-021-00181-2
• 发表时间: 2021
• 期刊: The Journal of membrane biology
• 作者: McDermott,JeffP;Sánchez,Gladis;Mitra,Amrita;Numata,September;Liu,LijunCatherine;Blanco,Gustavo
• 通讯作者: Blanco,Gustavo