SELF-MASKED ALDEHYDES AS INHIBITORS OF THE CYSTEINE PROTEASES 3CL PROTEASE, CATHEPSIN L, AND CRUZAIN

自掩蔽醛作为半胱氨酸蛋白酶 3CL 蛋白酶、组织蛋白酶 L 和 CruzAIN 的抑制剂

• 批准号: 10519117
• 负责人: Thomas Meek
• 金额: $ 19.69万
• 依托单位: TEXAS A&M AGRILIFE RESEARCH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-11-04 至 2024-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10519117
• 关键词: 2019-nCoV; Active Sites; Address; Aldehydes; Animal Model; Anti-Infective Agents; Autopsy; Binding; Binding Proteins; Biological Assay; COVID-19; COVID-19 pandemic; COVID-19 patient; COVID-19 treatment; Caspase; Cathepsin L; Cell model; Cells; Central America; Cessation of life; Chagas Disease; Communicable Diseases; Cysteine; Cysteine Proteinase Inhibitors; Drug Kinetics; Drug Targeting; Enzyme Inhibitor Drugs; Enzymes; Evaluation; Homologous Gene; Human; Individual; Infection; Lung; Maintenance; Malaria; Mammalian Cell; Masks; Medical; Metabolic; Microsomes; Modification; Parasites; Peptide Hydrolases; Peptides; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phase II Clinical Trials; Plasma; Play; Prodrugs; Property; Proteins; Publications; Reporting; Risk; Role; SARS-CoV-2 infection; SARS-CoV-2 inhibitor; SARS-CoV-2 variant; Sampling; Series; Severities; Solubility; South America; Structure; Therapeutic Agents; Time; Toxic effect; Trypanocidal Agents; Trypanosoma cruzi; Vaccines; aqueous; covalent bond; cruzain; design; human disease; improved; in vitro Assay; inhibitor; molecular modeling; nanomolar; novel; novel therapeutics; oxidation; scaffold

PROJECT SUMMARY Cysteine proteases play essential roles in the causative agents of numerous infectious diseases, including malaria, Chagas disease (CD, caused by the parasite Trypanosoma cruzi), and COVID- 19 (caused by the coronavirus SARS-CoV-2). There are neither vaccines nor well-tolerated therapies available for CD, which results in 50,000 annual deaths in Central and South America, with an estimated 300,000 infected individuals in the US. Cruzain is a cysteine protease that is essential to the establishment and maintenance of human infection by T. cruzi. By February 2021, the COVID-19 pandemic has resulted in 103 million cases worldwide and 2.2 million deaths. Despite the recent arrival of effective vaccines, the emergence of new variants of SARS-CoV-2 may render them less effective over time. Accordingly, the discovery of new therapeutic agents to treat COVID-19 remains a critical, unmet medical need. We recently showed that K11777, an irreversible, covalent inactivator of both cruzain and cathepsin L, potently blocked SARS-CoV-2 infection of mammalian cells by inactivation of the cysteine protease cathepsin L. Consequently, since then K11777 has advanced to Phase 2 clinical trials for the treatment of COVID-19. A recent publication reported that cathepsin L was significantly upregulated in lung autopsy samples from COVID-19 patients, suggesting that this CP not only has a key role in CoV-2 cell entry but also in the severity of human disease. Infection by SARS-CoV-2 requires the action of coronaviral protease 3CL protease (3CL-PR); also a cysteine protease, and consequently is an important drug target for COVID-19. Our approach is to identify inhibitors of cysteine proteases which form reversible covalent bonds with the invariant active-site cysteines of these enzymes. Ideally, selection of an appropriate peptide scaffold for such an inhibitor would provide a compound that potently inhibits more than one CP. Aldehydes are exceptionally potent, covalent, but reversible, inhibitors of cysteine proteases, despite the risk associated with their reactivity and metabolic instability. Here, we seek to re-address aldehydes as potent, reversible enzyme inhibitors by modifying them as intramolecular lactols, or “self-masked” aldehydes (SMAIs). The objective of this proposal is to design, synthesize, and evaluate new SMAIs, including prodrug forms, as novel inhibitors of these three CP drug targets, and evaluate their anti-infective properties in cellular models of CD and COVID-19. We will improve the drug-like properties of these new inhibitors in terms of pharmacokinetics, metabolic stability, and identify additional series of SMAIs. We will utilize prodrug forms of these inhibitors that provide enhanced metabolic stability from which active inhibitors should be released intercellularly upon enzymatic modification.

项目摘要 半胱氨酸蛋白酶在统一的传染病中起着重要作用， 包括疟疾，Chagas病（CD，由Cruzi引起的CD）和COVID- 19（由冠状病毒SARS-COV-2引起）。既没有疫苗也没有耐受性良好 可用于CD的疗法，在中美洲和南美导致每年50,000例死亡， 在美国，估计有300,000名受感染的人。 Cruzain是半胱氨酸蛋白酶 Cruzi T. Cruzi对人类感染的建立和维持至关重要。到2021年2月， COVID-19大流行已导致全球1.03亿例病例和220万例死亡。 尽管有效疫苗最近到来，但SARS-COV-2的新变体的出现 随着时间的流逝，它们的效率降低了。彼此之间，发现新的治疗剂 治疗Covid-19仍然是至关重要的，未满足的医疗需求。我们最近表明K11777，一个 Cruzain和组织蛋白酶L的不可逆转，共价灭活剂可能阻止SARS-COV-2 通过失活半胱氨酸蛋白结蛋蛋白L。因此，哺乳动物细胞感染。 从那时起，K11777已晋升为2期临床试验，以治疗Covid-19。最近 出版物报道，在肺尸检样品中，组织蛋白酶L已大大更新 COVID-19患者，表明该CP不仅在COV-2细胞进入中具有关键作用，而且在 人类疾病的严重程度。 SARS-COV-2感染需要冠状病毒的作用 蛋白酶3Cl蛋白酶（3Cl-pr）;还有半胱氨酸蛋白酶，因此是重要的 COVID-19的药物靶标。我们的方法是识别形成的半胱氨酸蛋白酶的抑制剂 与这些酶的不变活性位点半胱氨酸的可逆共价键。理想情况下， 选择适合这种抑制剂的合适肽支架将提供一种化合物 潜在抑制多个CP。醛具有极强的潜力，共价但可逆， 半胱氨酸蛋白酶的抑制剂，承担与其反应性和代谢相关的风险 不稳定。在这里，我们试图将醛重新用作潜力，可逆的酶抑制剂 将它们修饰为分子内乳糖，或“自我掩盖”醛（SMAIS）。目的 该建议是设计，合成和评估新的SMAI，包括前药形式，作为新颖的形式 这三个CP药物靶标的抑制剂，并评估其在细胞中的抗感染特性 CD和COVID-19的模型。我们将提高这些新抑制剂的类似药物样特性 药代动力学，代谢稳定性的术语，并确定了其他系列SMAI。我们将 利用这些抑制剂的前药形式，可提供增强的代谢稳定性 活性抑制剂应在酶促修饰后间间释放。