Integrated fragment-based phenotypic screening and chemoproteomics for identification of novel small cell lung cancer-specific targets

基于片段的表型筛选和化学蛋白质组学相结合，用于鉴定新型小细胞肺癌特异性靶标

• 批准号: 10577507
• 负责人: Andrii Monastyrskyi
• 金额: $ 23.63万
• 依托单位: H. LEE MOFFITT CANCER CTR & RES INST
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-02 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10577507
• 关键词: ASCL1 gene; Accounting; Affinity; Alkynes; Apoptosis; Azides; BETA2 protein; Binding; Binding Sites; Biochemical; Biological; Biological Assay; CASP3 gene; Cancer Etiology; Cancer Patient; Cancer cell line; Cell Line; Cell Survival; Cells; Cessation of life; Characteristics; Chemicals; Chemistry; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Copper; Data; Databases; Dedications; Diazomethane; Disease; Dose; Drug Costs; Drug Targeting; Epithelium; Future; Genetic; IGF1R gene; Immunotherapy; Induction of Apoptosis; Informatics; Knowledge; Lead; Libraries; Lung; Malignant Neoplasms; Malignant neoplasm of lung; Maps; Mass Spectrum Analysis; Mediating; Mining; Minor; Mission; Mutate; Non-Small-Cell Lung Carcinoma; Outcome; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phenotype; Proteins; Proteomics; Public Health; RB1 gene; RNA Interference; Research; Risk; Role; Signal Transduction; Structure; TP53 gene; Technology; Therapeutic; Time; Translating; Treatment Failure; Tumor Suppressor Proteins; United States National Institutes of Health; Validation; Western Blotting; analog; cancer cell; cancer subtypes; cancer therapy; chemical association; chemoproteomics; chemotherapy; cycloaddition; cytotoxic; driver mutation; drug development; drug discovery; drug resistance development; druggable target; improved; in vivo; inhibitor; innovation; insight; loss of function mutation; lung cancer cell; molecular subtypes; new therapeutic target; novel; novel therapeutics; patient population; response; scaffold; screening; small cell lung carcinoma; small molecule; standard of care; success; targeted treatment; transcription factor

PROJECT ABSTRACT Despite major successes in lung cancer therapy over the last two decades, there are still many patients who do not receive any benefit from targeted or immunotherapies. This is particularly true for small cell lung cancer (SCLC), a recalcitrant cancer for which there are currently no effective targeted therapies. Although SCLC patients initially respond well to conventional chemo- and immune therapy, drug resistance develops rapidly illustrating the need for new therapeutic targets and drugs. Traditional drug discovery often prioritize known targets and operates in limited biological and chemical space. Smaller fragment-like compounds are significantly better suited to enter previously unrecognized binding sites thereby probing uncharted biological target space and identifying novel targets. We hypothesize that cell-based screening of fragment-like probe molecules combined with chemoproteomics for target identification will enable the simultaneous identification of (i) novel, highly SCLC- specific targets and (ii) new lead compounds as modulators of these targets. We propose to specifically interrogate SCLC cells in comparison to NSCLC and non-cancerous cells and screen a library of small fragment-like molecules. Combination with innovative proteomics technology can subsequently identify new actionable targets, which will be functionally validated and will serve as starting points for novel SCLC-specific drug discovery projects. Aim 1: To identify actionable targets and associated chemical hit compounds in SCLC vs NSCLC cancer cells. We will screen a 15,000 fragment-like compound library (~estimated to the equivalent of >100,000 lead-like compounds) in several SCLC, which represent different molecular subtypes, and control (NSCLC and non-cancerous lung epithelial) cell lines. The most SCLC-specific or SCLC subtype-specific hits will be validated regarding selective viability and apoptosis effects, also in combination with standard of care chemotherapy drugs. Aim 2: To identify and validate novel and selective, actionable targets in SCLC cells. Using a diverse panel of fragment-like compounds with ‘privileged’ BioCore motifs and SCLC-selective screening hits from Aim 1 we will perform mass spectrometry-based chemoproteomics through UV-mediated photocrosslinking and copper-catalyzed ‘click’ chemistry in SCLC cell lines. High confidence protein target candidates, based on comparison with control compounds/cell lines and DEPMAP database mining, will be validated using appropriate biochemical and signaling readouts, as well as RNAi- and CRISPR-based genetic modulation in an expanded cell line panel. Medicinal chemistry optimization of hits will be done by 1) chemoinformatic and structure-network similarity analysis and 2) synthesis of analogs based on the SAR of both library and synthesized compounds. Leads will be characterized using biochemical and cell-based binding assays (e.g. affinity-immunoblotting, SPR, nanoBRET). The expected outcome is the identification of novel SCLC- and/or SCLC subtype-specific targets and chemical starting points for dedicated drug discovery efforts that through further elucidation of the biological context and in vivo validation produce desperately needed new therapeutics for SCLC.

项目摘要 尽管在过去的二十年里，肺癌治疗取得了重大成功，但仍有许多患者 没有从靶向或免疫治疗中获得任何益处。对于小细胞肺癌尤其如此 小细胞肺癌（SCLC）是一种恶性肿瘤，目前尚无有效的靶向治疗方法。虽然SCLC 患者最初对常规化疗和免疫治疗反应良好，耐药性迅速发展 说明了对新的治疗靶点和药物的需求。传统的药物发现往往优先考虑已知的 在有限的生物和化学空间内瞄准和运作。较小的碎片状化合物明显 更适合于进入以前未识别的结合位点，从而探测未知的生物靶空间， 确定新的目标我们假设，基于细胞的片段样探针分子筛选结合 利用化学蛋白质组学进行靶点鉴定将能够同时鉴定（i）新的、高度SCLC- 特异性靶点和（ii）作为这些靶点调节剂的新先导化合物。我们建议特别审问 SCLC细胞与NSCLC和非癌细胞的比较，并筛选小片段样 分子。与创新的蛋白质组学技术相结合，可以随后确定新的可操作的 这些靶点将在功能上得到验证，并将作为新型SCLC特异性药物的起点 发现项目。目的1：确定SCLC与 NSCLC癌细胞。我们将筛选15，000个片段样化合物文库（~估计相当于 > 100，000种铅样化合物），其代表不同的分子亚型，并且对照 （NSCLC和非癌性肺上皮）细胞系。最具SCLC特异性或SCLC亚型特异性的命中 将结合标准护理对选择性活力和细胞凋亡效应进行验证 化疗药物目的2：在SCLC细胞中鉴定和验证新的、选择性的、可操作的靶标。使用 一组具有“特权”BioCore基序和SCLC选择性筛选命中的片段样化合物 从目标1开始，我们将通过紫外光介导的光交联进行基于质谱的化学蛋白质组学 和铜催化的“点击”化学在SCLC细胞系。高置信度蛋白质靶候选物，基于 与对照化合物/细胞系和DEPMAP数据库挖掘的比较，将使用适当的 生物化学和信号读出，以及基于RNAi和CRISPR的遗传调节，在一个扩展的 细胞系面板。药物化学优化的命中将进行1）化学信息学和结构网络 相似性分析和2）基于库和合成化合物的SAR合成类似物。 将使用生物化学和基于细胞的结合测定（例如亲和免疫印迹，SPR， nanoBRET）。预期的结果是鉴定新的SCLC和/或SCLC亚型特异性靶点 和化学的起点，专门的药物发现工作，通过进一步阐明的生物 背景和体内验证产生了迫切需要的SCLC新疗法。