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.

抽象的 在超过 20% 的所有癌症和 50% 的乳腺癌，尤其是三阴性乳腺癌 (TNBC)。然而，尽管采取了广泛的努力来抑制 Wnt/-catenin 信号传导，目前尚未批准针对 Wnt 通路的药物，主要是由于无法 抑制剂药物将 β-连环蛋白的致癌性与稳态功能分开。 BCL9 是一种转录共 调节致癌β-连环蛋白介导的基因反式激活的激活剂。它的过表达增强了 Wnt/β- 连环蛋白信号传导，导致细胞增殖、迁移和体外侵袭，并诱导肿瘤发生 和体内进展。小鼠肠上皮细胞中 Bcl9/Bcl9l 的基因消融可减少肿瘤生长 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 假设证明是一种安全有效的癌症治疗策略。 Sapience Therapeutics 试图开发一种 -连环蛋白拮抗剂肽，旨在与 -连环蛋白相互作用。 我们的方法将氨基酸 (AA) 替换引入天然 HD2 结合域序列以增强 目标亲和力，包括用于细胞内进入的短细胞穿透结构域 (CPD)。另外，为了克服 针对传统肽遇到的挑战，Sapience的肽包括D-对映体AA（D-AAs）， 显着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，我们筛选了一个肽库，旨在 破坏 β-连环蛋白与 BCL9 的致癌关联，并选择 ST316 作为主要候选者。 ST316显示器 有效且选择性地抑制 Wnt 信号传导并诱导依赖于 Wnt 的肿瘤细胞的细胞毒性 信号传导（但不是 Wnt 独立肿瘤）。此外，ST316 表现出稳定性、优异的生物利用度和 溶解度和体内效力。在此 SBIR 的第一阶段中，ST316 表现出与 β- 的纳摩尔结合亲和力 连环蛋白和低微摩尔 (2.7μM) EC50 值可抑制体外 β-连环蛋白转录活性。安全 50mg/kg 剂量下没有组织病理学结果的研究表明治疗窗口至少为 10 倍。 根据这一概念验证，该 II 期申请提出了严格的 IND 支持毒理学和药物 支持 ST316 临床开发的宏观学研究。具体目标#1将确定首个人类 (FIH) ST316 剂量，通过确定大鼠中的无不良反应水平 (NOAEL) 和最高非严重毒性剂量 (HNSTD) 在小型猪中进行的 28 天重复剂量 GLP 毒性研究。在具体目标 #2 中，我们提出了一系列 确定小鼠患者来源异种移植物 (PDX) 中 ST316 药理活性剂量 (PAD) 的实验 模型，并识别与 PAD 相关的药效 (PD) 生物标志物，可用于临床 评估 ST316 活性。然后，PAD 将用于特定目标 #3，以评估 ST316 对 肿瘤免疫微环境（TME），即巨噬细胞极化、肿瘤浸润淋巴细胞（TIL） 肿瘤中的表达以及与 PD-1 抑制剂联合使用的影响。