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) 引起极其常见的皮肤和皮肤感染特定“高危”HPV 基因型的粘膜上皮感染可进展至前期。称为不典型增生的恶性病变，经过数年的时间，最终会导致侵袭性和侵袭性我们的治疗目标是治疗转移性上皮恶性肿瘤。子宫颈、生殖器和肛门在发展为侵入性疾病之前困扰着数百万妇女和男子癌症。 Kovina Therapeutics 的策略是通过选择性靶向 E6 来消除 HPV 感染 E6 蛋白并防止与其细胞结合伙伴相互作用对于病毒是必需的。基因组复制和维持，并且总是在 HPV 相关的发育不良和我们对化学库中的小分子进行了计算机筛选。类似于 E6 与其几个细胞伙伴的界面。 E6 与泛素连接酶 E6AP 的蛋白质-蛋白质相互作用，介导泛素化和使用该模型，我们使用蛋白酶体介导的肿瘤抑制蛋白 p53 的破坏。设计和合成的化合物 1) 与 HPV-16 E6 结合，2) 配备“弹头” 与 HPV-16 E6 结合口袋中的特定半胱氨酸形成共价键，形成 E6AP。方法来识别配备有反应弹头的分子，这些弹头可以介导共价和与半胱氨酸不可逆结合的例子有几种新的临床可用药物，这些药物靶向我们发现了几种共价结合的化合物。通过这个半胱氨酸残基连接到E6，阻断E6与E6AP的相互作用，恢复P53功能，诱导 HPV-16 表达细胞的死亡，包括 X 射线晶体学在内的多种生化分析。证明这些抑制剂占据目标 E6 口袋并与该半胱氨酸形成单一加合物。以共晶数据为指导的迭代优化可实现超过 80 种新颖的设计和合成化合物并产生了活性增强的化学型。提案计划筛选我们现有的 E6 抑制剂系列以确定其生物活性基于细胞的测定和药代动力学特性，符合 FDA 局部用药指南我们将启动药物开发研究，包括局部应用的配方和制剂。早期药代动力学和毒理学表征将得到第一阶段研究的成功。验证这一抗病毒策略并使我们能够继续申请 II 期 SBIR 来执行支持 IND 的研究。