Small Molecule E6 Inhibitors to Treat Dysplasia Caused by HPV Infections

小分子 E6 抑制剂治疗 HPV 感染引起的发育异常

基本信息

批准号：
10390563
负责人：
ELLIOT J. ANDROPHY
金额：
$ 29.85万
依托单位：
KOVINA THERAPEUTICS INC.
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10390563
关键词：
Animal Model Antiviral Agents Anus Apoptosis Applications Grants Attention Binding Biochemical Biological Biological Assay Cells Cervical Cervix Uteri Cessation of life Clinical Collection Complex Coupled Cream Crystallization Cysteine Data Drug Kinetics Drug Targeting Dysplasia Epithelial Evaluation Formulation Gel Genital Genitalia Genotype Goals Guidelines HIV Human Human Papilloma Virus Vaccine Human Papillomavirus Human papilloma virus infection Human papillomavirus 16 In Vitro Infection Infectious Skin Diseases Inflammatory Response Investigation Lead Lesion Liver Microsomes Lotion Maintenance Malignant - descriptor Malignant Neoplasms Malignant neoplasm of cervix uteri Measures Mediating Medical Molecular Weight Morbidity - disease rate Mucous Membrane Mus Neoplastic Cell Transformation Ointments Operative Surgical Procedures Pain Pharmaceutical Preparations Pharmacotherapy Phase Population Positioning Attribute Procedures Process Property Proteins Safety Series Skin Small Business Innovation Research Grant Small Molecule Chemical Library Solubility Specific qualifier value TP53 gene Testing Therapeutic Topical agent Topical application Toxic effect Toxicology Tumor Suppressor Proteins UBE3A gene Ubiquitination Vagina Viral Genome Virus Virus Diseases Vulva Wait Time Woman Work X-Ray Crystallography adduct base cancer invasiveness cellular targeting covalent bond design drug development experimental study high risk in silico in vivo inhibitor intraepithelial men metabolic abnormality assessment model design model development multicatalytic endopeptidase complex novel pharmacokinetic model phase 1 study premalignant prevent prophylactic protein protein interaction prototype screening small molecule small molecule libraries standard of care success systemic toxicity ubiquitin ligase

项目摘要

Human papillomaviruses (HPV) cause exceedingly common infections of cutaneous and mucosal epithelia. Infection with specific “high risk” HPV genotypes can progress to pre- malignant lesions called dysplasias, which over a period of years, can eventuate in invasive and metastatic epithelial malignancies. Our therapeutic goal is to treat early stage infections of the cervix, genitalia, and anus that afflict millions of women and men before these progress to invasive cancers. Kovina Therapeutics strategy is to eliminate HPV infection by selectively targeting the E6 protein and preventing interactions with its cellular binding partners. E6 is necessary for viral genome replication and maintenance and is always expressed in HPV-associated dysplasia and malignancies. We performed in silico screening of chemical libraries for small molecules that would resemble the interface of E6 with several of its cellular partners. Our assays focused on the protein-protein interaction of E6 with the ubiquitin ligase E6AP, which mediates ubiquitination and proteasome mediated destruction of the tumor suppressor protein p53. Using this model, we designed and synthesized compounds that 1) bind to HPV-16 E6 and 2) are armed with a ‘warhead’ to make a covalent bond with a specific cysteine in HPV-16 E6 binding pocket for E6AP. The approach to identify molecules equipped with reactive warheads that mediate covalent and irreversible binding to cysteine is exemplified in several new clinically available drugs that target proteins previously considered undruggable. We discovered several compounds covalently bound to E6 via this cysteine residue, block E6 interaction with E6AP, restore P53 function, and induce death of HPV-16 expressing cells. Multiple biochemical analyses including X-ray crystallography prove these inhibitors occupy the targeted E6 pocket and form a single adduct with this cysteine. Iterative optimization guided by the co-crystal data enabled design and synthesis of >80 novel compounds and has resulted chemotypes with increased activity. This one-year Phase I SBIR proposal plans to screen our existing E6 inhibitor collection to determine their biological activities in cell-based assays and pharmacokinetic properties that would meet FDA guidelines for a topical agent. We will initiate drug development studies including formulation for topical application and early stage pharmacokinetic and toxicology characterization. The success of Phase I study will validate this antiviral strategy and position us to proceed to apply for a Phase II SBIR to perform IND-enabling studies.

人乳头瘤病毒（HPV）引起皮肤和粘膜上皮。特定“高风险” HPV基因型的感染可以发展为前称为发育不良的恶性病变，多年来，这种病变可能会在侵入性和转移性上皮恶性肿瘤。我们的理论目标是治疗早期感染宫颈，生殖器和肛门在这些进展之前影响数百万的男女癌症。 Kovina Therapeutics策略是通过选择性靶向E6来消除HPV感染蛋白质并防止其与其细胞结合伴侣相互作用。 E6对于病毒是必需的基因组复制和维护，并且始终在HPV相关的发育不良和恶性肿瘤。我们在化学文库的计算机筛选中为小分子进行了筛选将类似于E6与其几个细胞伴侣的接口。我们的测定重点是 E6与泛素连接酶E6AP的蛋白质 - 蛋白质相互作用，介导泛素化和蛋白酶体介导的肿瘤抑制蛋白p53的破坏。使用此模型，我们 1）与HPV-16 E6结合的设计和合成化合物，配有“弹头” 在HPV-16 E6结合袋中与特定半胱氨酸建立共价键，用于E6AP。这识别与反应性弹头相等的分子的方法在几种针对的新临床可用药物中，与半胱氨酸的不可逆结合体现了蛋白质以前认为不良。我们发现了几种共价绑定的化合物通过此半胱氨酸居住地到E6，E6与E6AP相互作用，恢复p53功能和影响 HPV-16表达细胞的死亡。多次生化分析，包括X射线晶体学证明这些抑制剂占据了靶向的E6口袋，并用这种半胱氨酸形成单个加合物。由共结晶数据启用设计和合成> 80新颖的迭代优化化合物并导致了化学型，其活性增加。这个一年的我sbir 提案计划筛选我们现有的E6抑制剂收集以确定其生物学活动在基于细胞的测定和药代动力学特性中，符合局部的FDA指南代理人。我们将启动药物开发研究，包括用于局部应用和早期药代动力学和毒理学特征。第一阶段研究的成功将验证这种抗病毒策略，并使我们继续申请II期SBIR进行执行辅助研究。