Small molecule enhancers of tumor immunity targeting the LPA5 GPCR

针对 LPA5 GPCR 的肿瘤免疫小分子增强剂

基本信息

批准号：
10669778
负责人：
CORINNE ELIZABETH AUGELLI-SZAFRAN
金额：
$ 81.02万
依托单位：
UNIVERSITY OF TENNESSEE HEALTH SCI CTR
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：
2022-08-01 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10669778
关键词：
Adoptive Transfer Allogenic Animals Antigens Antitumor Response Biological Assay Breast Breast Carcinoma C57BL/6 Mouse CD8-Positive T-Lymphocytes Cancer Patient Cellular Assay Cytotoxic T-Lymphocytes Data Development Disinhibition Docking Drug Kinetics Enhancers Enzymes G-Protein-Coupled Receptors Generations Genetic Goals Growth Human Immune Evasion Immune system Immunologic Surveillance Immunomodulators Immunooncology Immunotherapy Impairment In Vitro Incidence Interleukin-2 Knock-out Knockout Mice Lead Liver Lymphocyte Lymphocytic Infiltrate Lymphoma Lysophospholipase Macrophage Malignant Epithelial Cell Malignant Neoplasms Malignant neoplasm of ovary Measures Metastatic Neoplasm to the Lung Methods Microsomes Modeling Molecular Mus Neoplasm Metastasis Outcomes Research Ovarian Ovum Pancreas Patients Peptides Pharmaceutical Chemistry Pharmaceutical Preparations Phenocopy Physiological Preclinical Testing Process Production Proliferating Property Publications Qualifying Receptor Activation Receptor Signaling Repression Research Research Personnel Resistance Role SUM-159 Breast Cancer Cell Line Solid Solubility Specificity Structure-Activity Relationship T cell response T-Cell Activation T-Cell Receptor T-Lymphocyte Testing Toxic effect Transforming Growth Factor alpha Transgenic Mice Tumor Immunity Tumor Promotion Wild Type Mouse acute toxicity analog animal efficacy antagonist anti-tumor immune response antigen-specific T cells beta-arrestin cancer cell cancer therapy cell killing clinical application cytotoxic cytotoxic CD8 T cells design drug discovery efficacy evaluation efficacy study high throughput screening human model immune checkpoint immune resistance immunological synapse formation in vivo indexing inhibitor iterative design lead candidate lead optimization lymphocyte proliferation lysophosphatidic acid melanoma meter mouse model neoplastic cell novel pharmacophore preclinical development receptor recruit refractory cancer response scaffold scale up secondary analysis small molecule small molecule inhibitor therapeutically effective tool tumor tumor microenvironment tumor progression virtual virtual screening

项目摘要

Lysophosphatidic acid (LPA) GPCR subtype 5 (LPAR5) is abundantly expressed by human and murine CD8 cytotoxic T lymphocytes (CTLs) and functions as an inhibitory receptor that represses T cell receptor (TCR) signaling leading to inhibition of tumor immunity. Specifically, stimulation of LPAR5 by physiological levels of LPA significantly impedes antigen specific TCR-induced Ca2+ mobilization, T cell activation, proliferation and cytolytic tumor cell killing functions, resulting in an impaired anti-tumor immune response. Indeed, CD8+ T cells lacking LPAR5 expression are more effective at reducing the growth rate of EG7 lymphoma and B16 melanoma tumors in mice compared to wild type (WT) CD8+ T cells. Moreover, Lpar5 -/- mice have 85% reduction in the incidence of B16 melanoma-derived lung metastasis compared to WT littermates, with a robust CD8+ CTL infiltration observed in the Lpar5 -/- mice that developed few lung metastasis. These data highlight a unique role for LPAR5 as an immune checkpoint molecule regulating immune surveillance and cytotoxic effector function. The objective of this proposal is to identify small molecule inhibitors of LPAR5 as clinically applicable immunomodulators for cancer treatment. A virtual screening (VS) of 2 million compounds using validated LPAR models identified more than 300 hits of which 90 were selective for LPAR5 antagonist compounds with diverse scaffolds. In addition, a high throughput screening (HTS) campaign of 200K compounds and secondary analyses of 19 validated hits have already resulted in the identification of two distinct molecular scaffolds. The most promising hit, SRI-42730, demonstrated LPAR5 antagonism in five independent assays: β-arrestin recruitment, Ca2+ mobilization, TGFα-shedding, IL-2 production implemented in HTS platform and in vivo efficacy in the B16 murine melanoma metastasis model. The novel hits and analogs we have already identified will be used as tool compounds in the following proposed studies: 1) Perform hit-to-lead medicinal chemistry optimization of LPAR5 antagonists. Computational approaches will include scaffold hopping on HTS hits, and the generation of a pharmacophore model to aid synthetic optimization of potency and selectivity of newly designed analogs; 2) Determine the specificity of novel antagonists at LPA GPCR subtypes and autotaxin lysophospholipase enzyme; 3) Rank specific antagonist hits by potency in boosting antigen-specific TCR activation and IL-2 production in the presence of LPA; 4). 8 key compounds will then be evaluated in cellular assays from which 3 compounds will undergo PK analysis prior to in vivo tox studies and animal efficacy studies. 5) Determine efficacy of nominated three potential lead compounds in boosting tumor immunity using murine and allogeneic human in vitro tumor killing assays and in vivo murine metastasis seeding and progression models. The impact of this research will be the identification and nomination of a single lead compound and a pool of structurally diverse LPAR5 antagonists for further preclinical development.

溶血磷脂酸 (LPA) GPCR 亚型 5 (LPAR5) 在人和鼠 CD8 中大量表达细胞毒性 T 淋巴细胞 (CTL) 并作为抑制性受体抑制 T 细胞受体 (TCR) 信号传导导致肿瘤免疫的抑制，特别是通过生理水平刺激 LPAR5。 LPA 显着阻碍抗原特异性 TCR 诱导的 Ca2+ 动员、T 细胞活化、增殖和 CD8+ T 细胞具有溶细胞性肿瘤细胞杀伤功能，导致抗肿瘤免疫反应受损。缺乏 LPAR5 表达更能有效降低 EG7 淋巴瘤和 B16 黑色素瘤的生长速度与野生型 (WT) CD8+ T 细胞相比，Lpar5 -/- 小鼠体内的肿瘤减少了 85%。与 WT 同窝小鼠相比，B16 黑色素瘤衍生肺转移的发生率，具有强大的 CD8+ CTL 在几乎没有发生肺转移的 Lpar5 -/- 小鼠中观察到的浸润这些数据强调了其独特的作用。 LPAR5作为调节免疫监视和细胞毒效应功能的免疫检查点分子。该提案的目的是确定临床适用的 LPAR5 小分子抑制剂使用经过验证的 200 万种化合物进行虚拟筛选 (VS)。 LPAR 模型鉴定了 300 多个命中，其中 90 个对 LPAR5 拮抗剂化合物具有选择性此外，还开展了 200K 化合物和二级化合物的高通量筛选 (HTS) 活动。对 19 个经过验证的命中的分析已经鉴定出两种不同的分子支架。最有希望的命中，SRI-42730，在五项独立测定中证明了 LPAR5 拮抗作用：β-arrestin HTS 平台和体内实施的招募、Ca2+ 动员、TGFα 脱落、IL-2 生产我们已经确定了 B16 小鼠黑色素瘤转移模型中的新药和类似物的功效。将在以下拟议研究中用作工具化合物： 1) 进行从命中到先导的药物化学 LPAR5拮抗剂的优化将包括HTS命中的支架跳跃，以及生成药效团模型以帮助合成优化新药的效力和选择性设计类似物；2) 确定新型拮抗剂对 LPA GPCR 亚型和自分泌运动因子的特异性溶血磷脂酶；3) 按增强抗原特异性 TCR 的效力对特定拮抗剂命中进行排序 LPA 存在下的激活和 IL-2 的产生；然后将在细胞中评估 8 种关键化合物。在体内毒性研究和动物功效研究之前，将对 3 种化合物进行 PK 分析 5) 确定指定的三种潜在化合物在增强肿瘤免疫力方面的功效。使用小鼠和同种异体人类体外肿瘤杀伤试验和体内小鼠转移播种和这一研究进展的影响将是单一先导的识别和提名。化合物和一组结构多样的 LPAR5 拮抗剂，用于进一步的临床前开发。