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：
10535248
负责人：
CORINNE ELIZABETH AUGELLI-SZAFRAN
金额：
$ 86.18万
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/10535248
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 system Immunologic Surveillance Immunomodulators Immunooncology Immunotherapy Impairment In Vitro Incidence Infiltration Interleukin-2 Knock-out Knockout Mice Lead Liver Lymphocyte Lymphocyte Function Lymphoma Lysophospholipase Malignant Epithelial Cell Malignant Neoplasms Malignant neoplasm of ovary Measures Metastatic Neoplasm to the Lung Methods Modeling Molecular Mus Neoplasm Metastasis Outcomes Research Ovarian Pancreas Patients Peptides Pharmaceutical Chemistry Pharmaceutical Preparations Physiological Preclinical Testing Process Production Property Publications Receptor Activation Receptor Signaling 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 Wild Type Mouse acute toxicity analog animal efficacy antagonist anti-tumor immune response antigen-specific T cells base beta-arrestin cancer cell cancer therapy cell killing clinical application cytotoxic cytotoxic CD8 T cells design drug discovery 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 lymphoid neoplasm lysophosphatidic acid macrophage melanoma 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）绝对由人和鼠CD8表达细胞毒性T淋巴细胞（CTL）和作为反映T细胞受体（TCR）的抑制受体起作用信号导致抑制肿瘤免疫学。具体而言，通过物理水平刺激LPAR5 LPA显着阻碍抗原特异性TCR诱导的Ca2+动员，T细胞激活，增殖和细胞溶解肿瘤细胞的杀伤功能，导致抗肿瘤免疫激发受损。确实，CD8+ T细胞缺乏LPAR5表达在降低EG7淋巴瘤和B16黑色素瘤的生长速率方面更有效与野生型（WT）CD8+ T细胞相比，小鼠的肿瘤。此外，LPAR5 - / - 小鼠的降低了85％与WT同窝仔相比，B16黑色素瘤来源的肺转移的发生率具有强大的CD8+ CTL 在发生肺转移很少的LPAR5 - / - 小鼠中观察到的浸润。这些数据突出了一个独特的角色对于LPAR5，作为调节免疫监测和细胞毒性效应子功能的免疫切除分子。该建议的目的是将LPAR5的小分子抑制剂识别为临床上适用用于癌症治疗的免疫调节剂。使用经过验证的200万种化合物的虚拟筛选（VS） LPAR模型确定了300多个命中，其中90个对LPAR5拮抗剂具有选择性潜水员脚手架。此外，200K化合物和次要的高吞吐量筛查（HTS）广告系列对19个验证命中的分析已经导致了两个不同的分子支架的鉴定。这最有希望的命中SRI-42730在五个独立测定中证明了LPAR5拮抗：β-arrestin 招聘，CA2+动员，TGFα脱落，IL-2生产在HTS平台和体内实施 B16鼠类黑色素瘤转移模型中的功效。我们已经确定的新颖的热门歌曲和类似物将在以下建议的研究中用作工具化合物：1） LPAR5拮抗剂的优化。计算方法将包括在HTS命中上跳脚手架，并且生成药效团模型，以帮助新近的效力合成优化设计的类似物； 2）确定LPA GPCR亚型和自身Xt蛋白的新型拮抗剂的特异性溶血磷脂酶； 3）通过促进抗原特异性TCR的效力来列为特定拮抗剂的命中 LPA存在的激活和IL-2产生； 4）。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效率之前，从3种化合物进行PK分析的测定法研究。 5）确定提名的三种潜在铅化合物在增强肿瘤免疫组织化学方面的效率使用鼠和同种异体人体体外肿瘤杀死测定法和体内鼠转移播种和进程模型。这项研究的影响将是单一潜在客户的识别和提名化合物和一组结构上不同的LPAR5拮抗剂，以进一步临床前开发。