Developing novel LOX inhibitors to target chemotherapy resistant TNBC

开发新型 LOX 抑制剂以靶向化疗耐药的 TNBC

• 批准号: 10696810
• 负责人: Ozgur Sahin
• 金额: $ 40.65万
• 依托单位: LOXIGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-08 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10696810
• 关键词: 3-Dimensional; Address; Antibody-drug conjugates; Benchmarking; Binding; Binding Proteins; Biological Assay; Biological Availability; Biometry; Breast Cancer Cell; Breast Cancer Model; Breast Cancer Patient; Breast Cancer cell line; Cancer Biology; Canis familiaris; Cardiotoxicity; Cell Line; Cells; Cessation of life; Chemoresistance; Chemosensitization; Collagen; Communication; Complement; Dedications; Doxorubicin; Drug Kinetics; Extracellular Matrix; Fibronectins; Goals; Grant; Hypoxia; Immunocompetent; Immunologic Deficiency Syndromes; Immunotherapy; In Vitro; Induction of Apoptosis; Lead; Legal patent; Libraries; Mammary Neoplasms; Maximum Tolerated Dose; Mediator; Medical Oncology; Metabolic; Mission; Modeling; Modification; Monoamine Oxidase; Mus; Nature; Oral; Organoids; Outcome; PTK2 gene; Patients; Penetration; Permeability; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacodynamics; Phase; Phenotype; Phosphotransferases; Plasma Proteins; Property; Protein-Lysine 6-Oxidase; Public Health; Reaction; Resistance; Safety; Serum Proteins; Signal Transduction; Small Business Technology Transfer Research; Solubility; Specificity; Structure-Activity Relationship; Testing; Therapeutic; Time; Toxic effect; United States National Institutes of Health; aggressive breast cancer; breast surgery; cancer subtypes; cellular targeting; chemosensitizing agent; chemotherapy; clinically relevant; crosslink; design; disability; drug candidate; drug discovery; effective therapy; genotoxicity; improved; in vitro Assay; in vivo; inhibitor; lead optimization; mRNA Expression; malignant breast neoplasm; member; meter; mortality; mouse model; novel; overexpression; patient derived xenograft model; pharmacokinetics and pharmacodynamics; pharmacophore; protein expression; screening; small molecule libraries; success; synergism; therapy resistant; translational potential; treatment strategy; triple-negative invasive breast carcinoma; tumor

PROJECT SUMMARY Triple negative breast cancer (TNBC) is the most aggressive breast cancer subtype. It accounts for ~15% of all breast cancer yet is responsible for 30% of breast cancer deaths. TNBC is treated primarily by conventional chemotherapy; however, resistance to therapy is common, leading to high mortality rates. Importantly, the benefit of current therapeutic strategies used in chemoresistant TNBC; i.e., immunotherapy and antibody- drug conjugates, is confined to only a fraction of patients, and survival benefit is limited. Therefore, there is an urgent need to identify novel and effective treatment strategies to overcome resistance to chemotherapy. Recently, we identified hypoxia-induced ECM re-modeler, lysyl oxidase (LOX) as a key mediator of chemoresistance in TNBC (Saatci et al, Nature Communications, 2020). We showed that LOX is overexpressed in chemoresistant tumors, and its inhibition re-sensitizes the most aggressive breast tumors to doxorubicin using several clinically-relevant mouse models. However, the available LOX inhibitors are either non-selective or has toxicity. Hence, our main objective in this project is to develop potent, specific and well-tolerated LOX inhibitors to overcome chemoresistance in TNBC that has a high translational potential. Through high-throughput compound library screening and hit-to-lead conversion studies, we identified compounds with potent on-target cellular engagement of LOX, with good oral pharmacokinetics (PK) and with chemosensitizer effect without major toxicity (US PTO 17/693,371 and PCT/US2022/20086, patent pending). Starting from our current non-optimized lead molecule, we aim to develop lead compounds with increased potency, safety and drug-likeness. To accomplish this goal, in Phase I of this Fast-Track STTR grant, we will generate a diverse library of small molecules via an extensive structure activity relationship (SAR) study using our initial pharmacophore. We will test the synthesized inhibitors with respect to the degree of LOX enzymatic activity inhibition, LOX binding and selectivity towards LOX. We will perform the off-target assessment of the inhibitors using CEREP screen as well as kinome profiling. The shortlisted candidates will further be tested in ECM crosslinking and 3D chemosensitization assays using both cell lines and organoids. Inhibitors with better efficacy, selectivity and stability will move to Phase II. In Phase II, we will perform several ADME assays, including metabolic stability/identity, Caco-2 permeability and transport, cardiotoxicity and genotoxicity, plasma protein binding, CYP inhibition/induction/reaction phenotyping to improve drug-like properties while maintaining on-target potency in TNBC cells. Detailed PK/PD and toxicity analyses of the most promising candidates will be carried out followed by testing their chemosensitizer effect using both state-of-the-art immunodeficient (cell line- and patient-derived xenografts) and immunocompetent (syngeneic) mice models. The successful completion of the proposed project will lead to potent and specific, lead-optimized LOX inhibitors to overcome chemoresistance in TNBC, the deadliest form of breast cancer.

项目总结 三阴性乳腺癌(TNBC)是最具侵袭性的乳腺癌亚型。它占到约15%的 然而，所有的乳腺癌都是导致30%的乳腺癌死亡的原因。TNBC主要由传统的 化疗；然而，对治疗的抵抗力很常见，导致高死亡率。重要的是， 目前用于化疗耐药的TNBC的治疗策略的好处；即免疫治疗和抗体- 药物结合物，仅限于一小部分患者，生存益处有限。因此，有 迫切需要确定新的和有效的治疗策略来克服对化疗的耐药性。 最近，我们发现低氧诱导的细胞外基质重塑因子赖氨酰氧化酶(LOX)是一种关键的细胞外基质调节因子。 TNBC的化疗耐药性(Saatci等人，自然传播，2020年)。我们证明了LOX是 在化疗耐药肿瘤中过度表达，其抑制使最具侵袭性的乳腺肿瘤重新敏感 使用几种临床相关的小鼠模型来研究阿霉素。然而，可用的LOX抑制剂是 非选择性或有毒性。因此，我们在这个项目中的主要目标是开发强有力的、具体的和 耐受性良好的LOX抑制剂，以克服具有高翻译潜力的TNBC的化疗耐药性。 通过高通量化合物文库筛选和Hit-to-Lead转化研究，我们确定了 具有较强的靶向LOX细胞结合力的化合物，具有良好的口服药物动力学(PK)和 具有化学增敏剂效应，没有重大毒性(US PTO 17/693,371和PCT/US 2022/20086，专利 待定)。从我们目前未优化的铅分子出发，我们的目标是开发具有以下特性的铅化合物 增强了效力、安全性和类似药物的特性。为了实现这一目标，在本快速通道STTR的第一阶段 格兰特，我们将通过广泛的结构活性关系建立一个不同的小分子文库 (SAR)使用我们最初的药效团进行的研究。我们将测试合成的缓蚀剂的程度 对LOX酶活性的抑制、LOX结合和对LOX的选择性。我们将进行偏离目标的表演 使用CEREP筛查和基因组图谱对抑制剂进行评估。入围的候选人将 进一步在使用细胞系和有机物的ECM交联和3D化疗增敏试验中进行测试。 药效、选择性和稳定性更好的抑制剂将进入第二阶段。在第二阶段，我们将进行几项 ADME分析，包括代谢稳定性/特性、Caco-2通透性和转运、心脏毒性和 遗传毒性、血浆蛋白结合、CYP抑制/诱导/反应表型以改善类药物 特性，同时在TNBC细胞中保持靶向效力。详细的PK/PD和毒性分析 最有希望的候选人将在随后使用这两种方法测试他们的化学增敏效果 最新的免疫缺陷(细胞系和患者来源的异种移植)和免疫活性 (同基因)小鼠模型。拟议项目的成功完成将带来强有力和具体的成果， 铅优化LOX抑制剂，以克服TNBC，最致命的乳腺癌形式的化疗耐药性。