Novel antibody-drug conjugates targeting T-cell lymphoma

靶向 T 细胞淋巴瘤的新型抗体-药物偶联物

• 批准号: 10386723
• 负责人: John P. Miller
• 金额: $ 24.65万
• 依托单位: ASYLIA THERAPEUTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10386723
• 关键词: Acids; Affinity; Antibodies; Antibody-drug conjugates; Antigen Targeting; Antigens; Antineoplastic Agents; Apoptotic; Attention; B-Lymphocytes; Binding; Biotinylation; Breast Cancer cell line; Camptothecin; Cathepsins; Cell Line; Cell Lineage; Cell Maturation; Cell Surface Proteins; Cell model; Cell surface; Cells; Characteristics; Clinic; Clinical; Conjugating Agent; Cutaneous; Cutaneous T-cell lymphoma; Data; Deacetylase; Development; Disease; Dose; Down-Regulation; Drug Kinetics; Drug toxicity; Exhibits; Failure; Feedback; Generations; Goals; Heat shock proteins; Heat-Shock Proteins 70; Hematologic Neoplasms; Heterogeneity; Human; Hypoxia; Immunotherapeutic agent; Investigational Drugs; Lead; Link; Literature; Lymphoma; Lymphoma cell; Malignant Neoplasms; Mass Spectrum Analysis; Membrane; Modeling; Molecular Chaperones; Monoclonal Antibodies; Multiple Myeloma; Mus; Mutation; Neoplasms; Normal tissue morphology; Patient-Focused Outcomes; Patients; Peripheral; Pharmaceutical Preparations; Pharmacology; Phase; Physiological; Preparation; Property; Proteome; Proteomics; Refractory; Regimen; Relapse; Resistance; Schedule; Series; Solid Neoplasm; Specificity; Surface; Surface Antigens; T-Cell Lymphoma; TNFRSF8 gene; Therapeutic; Therapeutic antibodies; Toxic effect; Toxicology; Tumor Antigens; Tumor Cell Line; Work; Xenograft procedure; anti-cancer; base; biological adaptation to stress; biomarker-driven; cancer cell; cell transformation; clinical translation; clinically relevant; cytotoxic; drug candidate; drug distribution; efficacy study; experience; experimental study; genome editing; human monoclonal antibodies; immunogenic cell death; improved outcome; in vivo; in vivo Model; inhibitor; novel; novel therapeutics; overexpression; patient derived xenograft model; peripheral blood; potential biomarker; pre-clinical; protein folding; pyrrolobenzodiazepine; research clinical testing; resistance mechanism; success; synergism; triple-negative invasive breast carcinoma; tumor; uptake

Abstract: T-cell lymphomas have remained treatment-challenging, not least because of their remarkable heterogeneity. The lab of Robert Orlovsky (co-founder of Asylia) analyzed the membrane proteome of peripheral and cutaneous T-cell lymphoma and discovered that surface Heat shock protein (csHSP)-70 as being consistently overexpressed. While HSP70 is well established as an intracellular stress-response chaperone, its localization to the cell surface in cancer, is gaining increased attention. Considering the recent clinical successes of antibody-drug conjugates and the exceptionally high and selective overexpression of cell surface HSP70 in T-cell lymphoma, we summarize that csHSP70 presents as an exceptionally attractive target for antibody drug conjugates (ADC) for the treatment of this malignancy. We have generated a high affinity monoclonal antibody (clone 239-87) specifically recognizing cell-surface HSP70. In preliminary data, we have generated an ADC of mAb 239-87 with MMAE, which specifically bound csHSP70 on T-cell NHL cell lines but did not bind normal peripheral blood-derived cells. Linked to MMAE, the 239-87-MMAE ADC showed cytotoxic activity against both peripheral and cutaneous T-cell lymphoma lines cell lines with comparable, or greater potency than the leading clinical ADC, brentuximab vedotin. We now propose a series of critical experiments to accelerate the clinical translation of 239-87 ADCs for the treatment of T-cell lymphoma. In aim 1, we will humanize mAb 239-87 and optimize the drug-like characteristics of the current murine clone. We will study binding of the leads to soluble HSP70, and to csHSP70 in wild-type and genome edited cell models to verify that they retain csHSP70 affinity and specificity. In specific aim 2, we will convert clone 239-87 mAb into an ADC. We will first synthesize a panel of ADCs based on the optimal lead antibody molecule linked to MMAE, a maytansine, a camptothecin, and a pyrrolobenzodiazepine with cathepsin- and acid-cleavable linkers and compare their efficacy against T-cell lymphoma cell lines. Next, we will genetically modulate csHSP70 and perform internalization and quantitative flow studies to determine the minimum csHSP70 expression needed to see anti-cancer efficacy. Finally, we will verify the potential for synergy between our ADCs and agents that enhance csHSP70, such as histione deacetylase inhibitors. In Aim 3, we will validate the activity of clone 239-87 ADCs using in vivo tumor cell line-derived models. Next, we will validate the activity of selected best performers in more physiologically relevant patient-derived xenograft (PDX) models. Finally, we will perform pharmacokinetic studies in these PDXs to begin to understand potential drug distribution and toxicity parameters. Taken together, these studies will provide a proof-of-concept that HSP70-targeted ADCs have the necessary efficacy and pharmacologic properties for development as attractive drug candidates and allow us to move forward with investigational new drug-enabling toxicology studies in preparation for phase I clinical testing.

摘要：T 细胞淋巴瘤的治疗仍然具有挑战性，尤其是因为其显着的 异质性。 Robert Orlovsky（Asylia 联合创始人）的实验室分析了外周血的膜蛋白质组 和皮肤 T 细胞淋巴瘤，并发现表面热休克蛋白 (csHSP)-70 持续过度表达。虽然 HSP70 已被广泛认为是细胞内应激反应伴侣，但它 癌症中细胞表面的定位正受到越来越多的关注。考虑到最近的临床成功 抗体-药物偶联物的研究以及细胞表面 HSP70 的极高选择性过表达 T 细胞淋巴瘤，我们总结 csHSP70 是抗体药物极具吸引力的靶点 结合物（ADC）用于治疗这种恶性肿瘤。我们已经产生了高亲和力单克隆抗体 （克隆 239-87）特异性识别细胞表面 HSP70。在初步数据中，我们生成了一个 ADC mAb 239-87 与 MMAE 特异性结合 T 细胞 NHL 细胞系上的 csHSP70，但不结合正常细胞 外周血来源的细胞。与 MMAE 相关，239-87-MMAE ADC 对两者都表现出细胞毒活性 外周和皮肤 T 细胞淋巴瘤系细胞系与领先的细胞系具有相当或更强的效力 临床 ADC，brentuximab vedotin。 我们现在提出一系列关键实验，以加速 239-87 ADC 的临床转化 T细胞淋巴瘤的治疗。在目标1中，我们将mAb 239-87人源化并优化药物样 当前小鼠克隆的特征。我们将研究引线与可溶性 HSP70 和 csHSP70 的结合 在野生型和基因组编辑的细胞模型中验证它们保留 csHSP70 亲和力和特异性。具体来说 目标 2，我们将克隆 239-87 mAb 转化为 ADC。我们将首先根据以下内容综合一组 ADC 与 MMAE、美登素、喜树碱和吡咯并苯二氮卓连接的最佳先导抗体分子 与组织蛋白酶和酸可裂解的接头结合，并比较它们对 T 细胞淋巴瘤细胞系的功效。下一个， 我们将对csHSP70进行基因调节并进行内化和定量流研究以确定 观察抗癌功效所需的最低 csHSP70 表达。最后，我们将验证协同作用的潜力 我们的 ADC 和增强 csHSP70 的药物（例如组氨酸脱乙酰酶抑制剂）之间的关系。在目标 3 中，我们将 使用体内肿瘤细胞系衍生模型验证克隆 239-87 ADC 的活性。接下来我们将验证 选定的最佳表现者在生理上更相关的患者来源的异种移植（PDX）模型中的活动。 最后，我们将对这些 PDX 进行药代动力学研究，以开始了解潜在的药物分布 和毒性参数。 总而言之，这些研究将提供一个概念证明，即针对 HSP70 的 ADC 具有 开发成为有吸引力的候选药物所需的功效和药理特性，使我们能够 推进新药毒理学研究，为第一阶段临床测试做准备。