SBIR-STTR Award

Development of Tumor and Immuno-Metabolism Based Small Molecule Therapeutics for Refractory Breast Cancer 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 IFN?, TNF?, IL-1? and decrease in tumor promoting TGF?, 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.

Public Health Relevance Statement:
Project Narrative Triple Negative Breast Cancer (TNBC) is defined by the receptor status of cancer cells. TNBC does not express estrogen, progesterone, or Her-2 receptors. TNBC is the most deadly sub- type of breast cancer, accounting for ~15% of breast cancer diagnoses and ~25% of breast cancer-related deaths. Median survival for 30% of the patients with TNBC is only one year. TNBC has poor clinical outcomes because it is highly metastatic, resistant to chemotherapy, and lacks targeted treatment options. TNBC cells are highly glycolytic, and therefore over- express monocarboxylate (lactate) transporters (MCTs) that are crucial for the cancer cells to adapt to glycolysis for their survival. In this proposal we outline the development of small molecule MCT1/4 dual inhibitors as a new therapeutic approach to effectively treat metastatic TNBC.

Project Terms:
4T1; Accounting; Achievement; Acute; Advanced Development; antitumor effect; Applications Grants; Back; base; Biological; Breast Cancer Cell; Breast Cancer cell line; breast cancer diagnosis; Breast Cancer Model; Breast Cancer Patient; cancer biomarkers; cancer cell; Cancer Cell Growth; cancer immunotherapy; Canis familiaris; CD8B1 gene; cell killing; Cell membrane; Cell Survival; Cells; Cessation of life; chemotherapy; Clinical; Clinical Trials; Combined Modality Therapy; Complement; cytokine; cytotoxicity; Data; design; Development; Development Plans; Diagnosis; Dose; Dose-Limiting; Drug Interactions; Drug Kinetics; effective therapy; Epigenetic Process; Estrogens; Evaluation; Event; Excretory function; experimental study; Extracellular Matrix; first-in-human; Glycolysis; Goals; Growth; Immune; immune activation; immune checkpoint blockade; immune function; Immunity; immunoregulation; Immunosuppression; in vivo; Inflammatory; inhibitor/antagonist; insight; Interferon Type II; Interleukin-1 beta; Interleukin-10; Lactate Transporter; macrophage; malignant breast neoplasm; Malignant Neoplasms; Maximum Tolerated Dose; MDA MB 231; Mediating; meetings; melanoma; Metabolic Pathway; Metabolism; Methods; Modeling; Molecular; Monkeys; mouse model; Mus; Mutation; novel; novel drug class; novel therapeutic intervention; novel therapeutics; Organ; Outcome; overexpression; Patients; Pharmaceutical Preparations; pharmacodynamic biomarker; Phase; Population; Positioning Attribute; pre-clinical; preclinical development; predictive marker; Process; Progesterone; Prognostic Marker; programs; Protein Family; Rattus; receptor; Refractory; Regimen; Relapse; Reporting; Resistance; Safety; safety study; scale up; Schedule; Signal Pathway; Small Business Innovation Research Grant; small molecule; small molecule therapeutics; standard of care; success; synergism; targeted agent; targeted treatment; Testing; TNF gene; Toxic effect; Toxicology; Transforming Growth Factor beta; transport inhibitor; triple-negative invasive breast carcinoma; tumor; tumor growth; Tumor Immunity; tumor metabolism; tumor microenvironment; tumor progression; tumor xenograft; Tumor-Infiltrating Lymphocytes; Validation; Woman; Xenograft Model; Xenograft procedure

Development of Tumor and Immuno-Metabolism Based Small Molecule Therapeutics for RefractoryBreast CancerWorldwide, approximately 1 million women are diagnosed with breast cancer each year. Triple Negative BreastCancer (TNBC) is defined as that which does not express estrogen, progesterone, or Her-2 receptors. TNBC isthe most deadly sub-type of breast cancer, accounting for ~15% of the breast cancer diagnoses and ~25% ofbreast cancer-related deaths. Median survival for 30% of the patients with TNBC is one year. TNBC has poorclinical outcomes due to its high metastatic rate, resistance to chemotherapy, and lack of effective treatmentoptions. Although immunotherapy for cancers is rapidly expanding with the discovery of new targets and methodsto activate immune function within tumors, it has only shown success in a limited subset of metastatic TNBCpatients. The lactate-rich TNBC tumor microenvironment (TME) has been shown to be highlyimmunosuppressive, promoting tumor growth and progression. Cancer cells transport lactate across the cellmembrane to the extracellular matrix via monocarboxylate transporters, MCT1 and MCT4. We have developeddual MCT1/4 inhibitors (dMCTi) to block lactate excretion to the TME thereby directly killing cancer cells andsimultaneously activating local immunity in the TME. In our preliminary studies, we have shown that dMCTi arepotent compounds against multiple TNBC cell lines. Also, we have shown that in in vivo experiments with bothmouse 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 efficacyin MDA-MB-231 immune-deficient xenograft model shows inhibitors' direct cell killing effect. In the 4T1 and SM1syngeneic models, we observed a decrease in expression of multiple immunosuppressive molecules such asB7 family proteins, macrophage polarization to M1, MDSCs, and increase in CD8+ population in treated tumorscompared to the control tumors. Furthermore, profiling of cytokines indicated an increase in pro-inflammatoryIFNγ, TNFα, IL-1β and decrease in tumor promoting TGFβ, IL-10 in treated tumors compared to the controltumors confirming that the anti-tumor effect of dMCTis is in part due to enhanced immune function. Supportedby these preliminary data, we selected a dMCTi, NGY-B, as a pre-clinical development candidate. In this DirectPhase-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, andrequest a pre-IND Type B meeting with FDA. Upon completion of these Aims, Nirogyone will have establishedNGY-B's scalability, broader efficacy, and potential dose-limiting toxicities. We will then submit a SBIR PhaseIIb grant application to evaluate non-GLP and GLP toxicology studies and assemble the IND package for first-in-human clinical trials.

Public Health Relevance Statement:
Project Narrative Triple Negative Breast Cancer (TNBC) is defined by the receptor status of cancer cells. TNBC does not express estrogen, progesterone, or Her-2 receptors. TNBC is the most deadly sub- type of breast cancer, accounting for ~15% of breast cancer diagnoses and ~25% of breast cancer-related deaths. Median survival for 30% of the patients with TNBC is only one year. TNBC has poor clinical outcomes because it is highly metastatic, resistant to chemotherapy, and lacks targeted treatment options. TNBC cells are highly glycolytic, and therefore over- express monocarboxylate (lactate) transporters (MCTs) that are crucial for the cancer cells to adapt to glycolysis for their survival. In this proposal we outline the development of small molecule MCT1/4 dual inhibitors as a new therapeutic approach to effectively treat metastatic TNBC.

Project Terms:
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