Development of a cell-penetrating beta-catenin antagonist peptide as a therapeutic candidate for Wnt-driven breast cancer

开发细胞穿透性 β-连环蛋白拮抗剂肽作为 Wnt 驱动乳腺癌的候选治疗药物

• 批准号: 10707593
• 负责人: Jim Rotolo
• 金额: $ 136.23万
• 依托单位: SAPIENCE THERAPEUTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-02 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10707593
• 关键词: Ablation; Advanced Development; Adverse effects; Affinity; Amino Acid Substitution; Amino Acids; Animal Testing; BCL9 gene; Binding; Biological Availability; Biological Markers; Breast; Breast Cancer Model; Breast Cancer Treatment; Cancerous; Cell Proliferation; Cell Survival; Cell physiology; Cells; Cessation of life; Clinical; Data; Development; Dose; Drug Targeting; Genes; Genetic; Genetic Transcription; Growth; Guidelines; Human; In Vitro; Infiltration; Intravenous infusion procedures; Lymphocyte; Macrophage; Malignant Neoplasms; Mediating; Metastatic Neoplasm to Lymph Nodes; Miniature Swine; Mus; Oncogenic; Pathway interactions; Patients; Penetration; Peptide Library; Peptides; Pharmaceutical Preparations; Pharmacodynamics; Pharmacology Study; Pharmacology and Toxicology; Phase; Phenotype; Prognosis; Proteins; Rattus; Recurrence; Research Design; Series; Signal Pathway; Signal Transduction; Small Business Innovation Research Grant; Solubility; Sprague-Dawley Rats; Structure-Activity Relationship; Testing; Therapeutic; Tissues; Toxic effect; Toxicokinetics; Toxicology; Transactivation; Transcription Coactivator; Tumor Escape; Tumor Immunity; Tumor-Infiltrating Lymphocytes; WNT Signaling Pathway; Xenograft procedure; antagonist; anti-tumor immune response; beta catenin; cancer therapy; chemotherapy; clinical development; clinically relevant; cytotoxicity; design; enantiomer; experimental study; first-in-human; immunogenicity; in vitro activity; in vivo; inhibitor; inhibitor therapy; intestinal epithelium; lead candidate; malignant breast neoplasm; migration; nanomolar; neoplastic cell; overexpression; participant enrollment; patient derived xenograft model; pharmacodynamic biomarker; pharmacologic; programmed cell death ligand 1; receptor; safety study; standard of care; success; therapeutic candidate; therapeutic target; triple-negative invasive breast carcinoma; tumor; tumor growth; tumor initiation; tumor microenvironment; tumor progression; tumor-immune system interactions

ABSTRACT Dysregulated overexpression and hyperactivation of Wnt/-catenin signaling is found in >20% of all cancers and in 50% of breast cancer, particularly triple negative breast cancer (TNBC). Yet despite extensive efforts to inhibit Wnt/-catenin signaling, no drugs that target the Wnt pathway have been approved, mainly due to the inability of inhibitors drugs to separate oncogenic from homeostatic functions of -catenin. BCL9 is a transcriptional co- activator that regulates oncogenic β-catenin-mediated gene transactivation. Its overexpression enhances Wnt/β- catenin signaling, resulting in cell proliferation, migration and invasiveness in vitro, and induces tumor initiation and progression in vivo. Genetic ablation of Bcl9/Bcl9l in mouse intestinal epithelium reduces tumor growth without phenotypic abnormalities. Therefore, specific antagonism of the interaction of β-catenin with BCL9 is hypothesized to prove a safe and efficacious strategy for cancer therapy. Sapience Therapeutics sought to develop a -catenin antagonist peptide, designed to interact with -catenin. Our approach introduces amino acid (AA) substitutions to the native HD2 binding domain sequence to enhance target affinity and includes a short cell penetrating domain (CPD) for intracellular entry. Additionally, to overcome challenges encountered by traditional peptides, Sapience’s peptides include D-enantiomer AAs (D-AAs), which dramatically increase stability and bioavailability, while reducing potential immunogenicity. Through extensive structure-activity relationship (SAR) studies, we screened a library of peptides designed to disrupt β-catenin’s oncogenic association with BCL9 and selected ST316 as the lead candidate. ST316 displays potent and selective inhibition of Wnt signaling and induction of cytotoxicity in tumor cells dependent on Wnt signaling (but not Wnt-independent tumors). Further, ST316 demonstrates stability, superior bioavailability and solubility and in vivo potency. In Phase I of this SBIR, ST316 demonstrated nanomolar binding affinity to β- catenin and a low micromolar (2.7µM) EC50 value for inhibition of β-catenin transcriptional activity in vitro. Safety studies with no histopathological findings at 50mg/kg suggested a therapeutic window of at least 10x. Following this proof of concept, this Phase II application proposes rigorous IND-enabling toxicology and phar- macology studies to support clinical development of ST316. Specific Aim #1 will determine the first-in-human (FIH) ST316 dose by identifying the no adverse effect level (NOAEL) in rats and highest non-severely toxic dose (HNSTD) in minipigs in 28-day repeat dose GLP toxicity studies. In Specific Aim #2, we propose a series of experiments to determine the ST316 pharmacologic active dose (PAD) in mouse patient-derived xenograft (PDX) models, and identify pharmacodynamic (PD) biomarkers associated with the PAD, which can be used clinically to assess ST316 activity. The PAD will then be used in Specific Aim #3 to evaluate the impact of ST316 on the tumor immune microenvironment (TME), namely macrophage polarization, tumor infiltrating lymphocyte (TIL) expression in tumors and the impact of combination with a PD-1 inhibitor.

摘要 Wnt/-catenin信号的异常过表达和过度激活在所有癌症和 在50%的乳腺癌中，尤其是三阴性乳腺癌(TNBC)。然而，尽管做出了广泛的努力来抑制 Wnt/-连环蛋白信号转导途径，目前还没有针对Wnt途径的药物被批准，主要是由于无法 用于分离-连环蛋白致癌和稳态功能的抑制剂药物。Bcl9是一种转录辅助性基因。 调节致癌β-连环蛋白介导的基因反式激活的激活剂。其过度表达增强了Wnt/β- 连环蛋白信号，在体外导致细胞的增殖、迁移和侵袭，并诱导肿瘤的启动 以及在体内的进展。基因消融小鼠肠上皮细胞Bcl9/Bcl9l抑制肿瘤生长 无表型异常。因此，β-连环蛋白与bcl9相互作用的特异性拮抗作用是 假设证明这是一种安全有效的癌症治疗策略。 Sapience Treateutics试图开发一种-连环蛋白拮抗剂多肽，旨在与-连环蛋白相互作用。 我们的方法在天然的HD2结合结构域序列中引入氨基酸(AA)取代以增强 靶向亲和力，包括用于细胞内进入的短细胞穿透域(CPD)。此外，要克服 传统多肽面临的挑战，Sapience的多肽包括D-对映体氨基酸(D-AAs)，它 显著提高稳定性和生物利用度，同时降低潜在的免疫原性。 通过广泛的构效关系(SAR)研究，我们筛选了一个设计用于 破坏β-连环蛋白与bcl9的致癌关联，并选择st316作为首选候选基因。ST316显示器 依赖Wnt的肿瘤细胞对Wnt信号的有效和选择性抑制及细胞毒作用的诱导 信号转导(但不是Wnt非依赖性肿瘤)。此外，ST316表现出稳定性、卓越的生物利用度和 溶解度和体内效力。在此SBIR的第一阶段，ST316显示了与β-的纳分子结合亲和力。 低微摩尔(2.7mM)EC50值用于体外抑制β-连环蛋白转录活性。安全问题 没有组织病理学发现的研究表明，50 mg/kg的治疗窗口至少是10倍。 根据这一概念证明，该第二阶段应用程序提出了严格的IND毒理学和药物- 支持ST316临床开发的眼压学研究。第一个具体目标将决定第一个人类 (FIH)ST316剂量，通过确定大鼠的无不良反应水平(NOAEL)和最高非严重毒性剂量 (HNSTD)在28天重复剂量的GLP毒性研究中的小型猪。在具体目标2中，我们提出了一系列 小鼠移植瘤ST316药理活性剂量(PAD)测定的实验研究 模型，并确定与PAD相关的药效学(PD)生物标记物，可用于临床 以评估ST316的活性。然后，PAD将用于特定的目标#3，以评估ST316对 肿瘤免疫微环境，即巨噬细胞极化、肿瘤浸润性淋巴细胞(TIL) 在肿瘤中的表达以及与PD-1抑制剂联合使用的影响。