Development of Tumor and Immuno-Metabolism Based Small Molecule Therapeutics for Refractory Breast Cancer

肿瘤与免疫代谢为主的小分子药物治疗难治性疾病的研究进展 乳腺癌 摘要 在世界范围内，每年约有100万妇女被诊断为乳腺癌。三重负面乳房 肿瘤(TNBC)是指不表达雌激素、孕激素或HER-2受体的肿瘤。TNBC是 最致命的乳腺癌亚型，约占乳腺癌诊断的15%，约25% 与乳腺癌相关的死亡。30%的TNBC患者的中位生存期为一年。TNBC很差 高转移率、化疗耐药和缺乏有效治疗导致的临床转归 选择。尽管随着新靶点和新方法的发现，癌症的免疫疗法正在迅速扩大 为了激活肿瘤内的免疫功能，它只在转移性TNBC的有限亚群中显示出成功。 病人。富含乳酸的TNBC肿瘤微环境(TME)已被证明是高度 免疫抑制，促进肿瘤生长和进展。癌细胞在细胞内转运乳酸 膜通过单羧酸转运体MCT1和MCT4进入细胞外基质。我们已经开发出 双MCT1/4抑制剂(DMCTi)可阻止乳酸向TME的排泄，从而直接杀死癌细胞和 同时激活TME的局部免疫力。在我们的初步研究中，我们已经表明dMCTi是 有效的化合物对多种TNBC细胞系有抑制作用。此外，我们已经证明，在活体实验中， 小鼠异种移植模型(MDA-MB-231，乳腺癌)和同基因黑色素瘤和TNBC小鼠模型； SM1(黑色素瘤，BRAFV600E)和4T1(TNBC)，dMCTi具有显著的抗肿瘤作用。抗肿瘤疗效 在MDA-MB-231免疫缺陷异种移植模型中显示了抑制剂的直接细胞杀伤作用。在4T1和SM1中 同基因模型，我们观察到多种免疫抑制分子的表达减少，如 治疗肿瘤中B7家族蛋白、巨噬细胞对M1、MDSCs的极化以及CD8+细胞群的增加 与对照组肿瘤相比。此外，细胞因子的分析表明促炎因子的增加 干扰素γ、肿瘤坏死因子α、白介素1β及肿瘤抑制促进转化生长因子β、白介素10的表达 肿瘤研究证实，dMCTis的抗肿瘤作用部分是由于增强了免疫功能。支撑点 根据这些初步数据，我们选择了一个dMCTI，NGY-B，作为临床前开发的候选者。在此直销中 第二阶段应用，我们建议(1)进行NGY-B的临床前药代动力学和安全性研究，(2) 建立NGY-B的有效剂量方案，研究NGY-B在几种小鼠肿瘤模型中的疗效。 和体内免疫抑制机制的研究，(3)建立NGY-B的可制造性，以及 要求与FDA进行一次B型IND前会议。在完成这些目标后，尼罗乔内将建立 NGY-B的可扩展性、更广泛的疗效和潜在的剂量限制毒性。然后，我们将提交SBIR阶段 IIB拨款申请，以评估非GLP和GLP毒理学研究，并组装IND包，用于第一次- 人体临床试验。

Development of Tumor and Immuno-Metabolism Based Small Molecule Therapeutics for Refractory Breast Cancer Abstract Worldwide, approximately 1 million women are diagnosed with breast cancer each year. Triple Negative Breast Cancer (TNBC) is defined as that which does not express estrogen, progesterone, or Her-2 receptors. TNBC is the most deadly sub-type of breast cancer, accounting for ~15% of the breast cancer diagnoses and ~25% of breast cancer-related deaths. Median survival for 30% of the patients with TNBC is one year. TNBC has poor clinical outcomes due to its high metastatic rate, resistance to chemotherapy, and lack of effective treatment options. Although immunotherapy for cancers is rapidly expanding with the discovery of new targets and methods to activate immune function within tumors, it has only shown success in a limited subset of metastatic TNBC patients. The lactate-rich TNBC tumor microenvironment (TME) has been shown to be highly immunosuppressive, promoting tumor growth and progression. Cancer cells transport lactate across the cell membrane to the extracellular matrix via monocarboxylate transporters, MCT1 and MCT4. We have developed dual MCT1/4 inhibitors (dMCTi) to block lactate excretion to the TME thereby directly killing cancer cells and simultaneously activating local immunity in the TME. In our preliminary studies, we have shown that dMCTi are potent compounds against multiple TNBC cell lines. Also, we have shown that in in vivo experiments with both mouse xenograft models (MDA-MB-231, breast cancer) and syngeneic mouse models of melanoma and TNBC; SM1 (melanoma, BRAFV600E), and 4T1 (TNBC), dMCTi exert significant anti-tumor efficacy. Anti-tumor efficacy in MDA-MB-231 immune-deficient xenograft model shows inhibitors’ direct cell killing effect. In the 4T1 and SM1 syngeneic models, we observed a decrease in expression of multiple immunosuppressive molecules such as B7 family proteins, macrophage polarization to M1, MDSCs, and increase in CD8+ population in treated tumors compared to the control tumors. Furthermore, profiling of cytokines indicated an increase in pro-inflammatory IFNγ, TNFα, IL-1β and decrease in tumor promoting TGFβ, IL-10 in treated tumors compared to the control tumors confirming that the anti-tumor effect of dMCTis is in part due to enhanced immune function. Supported by these preliminary data, we selected a dMCTi, NGY-B, as a pre-clinical development candidate. In this Direct Phase-II application, we propose to (1) conduct preclinical pharmacokinetic and safety studies of NGY-B, (2) Establish an effective dose regimen of NGY-B, investigate the efficacy of NGY-B in several mouse tumor models, and study the immune suppressive mechanisms in vivo, (3) establish NGY-B scalability for manufacturing, and request a pre-IND Type B meeting with FDA. Upon completion of these Aims, Nirogyone will have established NGY-B’s scalability, broader efficacy, and potential dose-limiting toxicities. We will then submit a SBIR Phase IIb grant application to evaluate non-GLP and GLP toxicology studies and assemble the IND package for first- in-human clinical trials.