KR-20260065705-A - Pharmaceutical composition for preventing or treating cancer, comprising an MTHFD2 expression or activity inhibitor as an active ingredient

Abstract

The present invention relates to a pharmaceutical composition for the prevention or treatment of cancer comprising an inhibitor of MTHFD2 expression or activity as an active ingredient. It was confirmed that inhibiting the expression of MTHFD2 can suppress the activity of cancer-associated fibroblasts, which play a key role in cancer growth and metastasis among the cells constituting the cancer microenvironment. Furthermore, various experiments confirmed that inhibiting the expression of MTHFD2 in cancer-associated fibroblasts significantly suppresses cancer cell growth and metastasis; thus, it is expected that the inhibition of cancer-associated fibroblast activity through the inhibition of MTHFD2 expression will be usefully utilized as a novel cancer treatment strategy.

Inventors

• 이상명
• 황예은

Assignees

• 중앙대학교 산학협력단

Dates

Publication Date

20260511

Application Date

20241101

Claims (17)

1. A pharmaceutical composition for the prevention or treatment of cancer comprising one or more active ingredients selected from the group consisting of the following: MTHFD2 (methylenetetrahydrofolate dehydrogenase 2) expression or activity inhibitor; Fibroblasts with inhibited MTHFD2 expression or activity; and Culture medium of fibroblasts with inhibited MTHFD2 expression or activity.
2. In paragraph 1, A pharmaceutical composition wherein the above MTHFD2 expression or activity inhibitor is an inhibitor that inhibits the expression or activity of MTHFD2 in cancer-associated fibroblasts (CAF).
3. In paragraph 1, A pharmaceutical composition characterized in that the above culture medium does not contain fibroblasts.
4. In paragraph 1, A pharmaceutical composition characterized in that the above culture medium has reduced expression levels of IL-6 and IL-8.
5. In paragraph 1, A pharmaceutical composition in which the above cancer is a solid tumor.
6. In paragraph 5, A pharmaceutical composition wherein the above cancer is one or more selected from the group consisting of lung cancer, non-small cell lung cancer, prostate cancer, colorectal cancer, liver cancer, pancreatic cancer, melanoma, squamous cell carcinoma, breast cancer, head and neck cancer, thyroid cancer, soft tissue sarcoma, osteosarcoma, testicular cancer, ovarian cancer, bladder cancer, skin cancer, brain cancer, angiosarcoma, mastocytoma, leukemia, lymphoma, gastric cancer, kidney cancer, hematopoietic tumor, neuroblastoma, epidermal carcinoma, and metastatic cancer thereof.
7. In paragraph 1, A pharmaceutical composition wherein the above expression or activity inhibitor is one or more selected from the group consisting of compounds, peptides, aptamers, polynucleotides, antibodies, recombinant vectors, antisense nucleotides, siRNA (small interference RNA), shRNA (short hairpin RNA), miRNA (microRNA), ribozymes, DNAzymes, PNA (peptide nucleic acids), antibodies, CRISPR, and aptamers.
8. In paragraph 1, The above composition is a pharmaceutical composition that inhibits the tumor microenvironment.
9. In paragraph 1, The above composition is a pharmaceutical composition that inhibits the invasion, growth, and metastatic activity of cancer cells.
10. In paragraph 1, A pharmaceutical composition wherein the above composition is characterized by one or more selected from the group consisting of the following: By reducing ROS levels induced by NAD + and inhibiting mitochondrial division, it reduces cytoplasmic mtDNA levels and inhibits cytokines by inhibiting the activity of the cGAS-STING pathway induced by mtDNA; and Reduces the activity and contractility of fibroblasts.
11. A pharmaceutical composition for inhibiting cancer metabolism, comprising, at an in vitro level, one or more active ingredients selected from the group consisting of the following: MTHFD2 (methylenetetrahydrofolate dehydrogenase 2) expression or activity inhibitor; Fibroblasts with inhibited MTHFD2 expression or activity; and Fibroblast culture medium with inhibited MTHFD2 expression or activity.
12. In Paragraph 11, A pharmaceutical composition characterized in that the above culture medium does not contain fibroblasts.
13. In Paragraph 11, A pharmaceutical composition characterized in that the above culture medium has reduced expression levels of IL-6 and IL-8.
14. A method for inhibiting cancer metabolism comprising the step of administering a composition comprising one or more active ingredients selected from the group consisting of the following: MTHFD2 (methylenetetrahydrofolate dehydrogenase 2) expression or activity inhibitor; MTHFD2-inhibiting fibroblasts; and MTHFD2 expression inhibiting fibroblast culture medium.
15. In Paragraph 14, A method for inhibiting cancer metabolism, characterized in that the above culture medium does not contain fibroblasts.
16. In Paragraph 14, A method for inhibiting cancer metabolism, characterized in that the above culture medium has reduced expression levels of IL-6 and IL-8.
17. A kit for preventing or treating cancer, comprising the composition of claim 1 and instructions.

Description

Pharmaceutical composition for preventing or treating cancer, comprising an MTHFD2 expression or activity inhibitor as an active ingredient The present invention relates to a pharmaceutical composition for the prevention or treatment of cancer comprising an MTHFD2 expression or activity inhibitor as an active ingredient. When considered as a single entity, cancer has become the second leading cause of death worldwide. The cost of cancer treatment is skyrocketing as demands on healthcare budgets increase across all countries. Cancer cells have a metabolism very different from normal cells; their higher metabolism allows them to maintain higher proliferation rates and resist certain cell death signals. The importance of cellular metabolism in cancer development was initially observed when Warburg discovered that tumor cells exhibited enhanced glycolytic activity compared to normal cells. This phenomenon, now known as the “Warburg effect,” has recently become the focus of intensive efforts in the discovery of new therapeutic targets and novel cancer drugs. However, cancer cells do not simply enhance glycolytic activity in metabolism by upregulating the expression of enzymes in the glycolytic pathway to facilitate numerous reactions within that pathway. It was observed that energy is primarily produced by lactic acid fermentation in the cytoplasm following a high rate of glycolysis, rather than by the oxidation of pyruvate following a relatively low rate of glycolysis in the mitochondria as in most normal cells. The former is anaerobic, which does not use oxygen, and the latter is aerobic, which uses oxygen. This transition in cancer cell metabolism occurs even in oxygen-rich environments. It was discovered that this transition occurs in cancer cell metabolism because the conversion of phosphoenolpyruvate to pyruvate, catalyzed by the pyruvate kinase enzyme, is not accelerated but rather weakened in cancer cells. This fact indicates that under aerobic conditions, cancer cells tend to prefer metabolism via glycolysis over the much more efficient oxidative phosphorylation pathway preferred by most other cells in the body, and it is known that pyruvate, the final product of glycolysis, is converted into lactic acid in tumor cells. Recently, there has been significant interest in developing anticancer therapies that target cell signaling pathways important for the metabolism and growth of cancer cells. Representative drugs involved in cancer cell metabolism include biguanide compounds such as metformin and 2-deoxy-D-glucose, but effective treatments have not yet been reported. Figures 1a to 1e show that, based on big data analysis results, the expression of MTHFD2 is increased in the stroma of solid tumors, including lung cancer. Figure 1a is a schematic diagram and heatmap showing the results of an analysis using mouse embryonic fibroblasts (MEFs) cultured in a soft environment (0.5 kPa polyacrylamide gel (PAG)) (cells were treated with 2 ng/ml TGF-β1 for 8 hours). As a result, differentially expressed genes (DEGs; upregulation: 316, downregulation: 290) were identified in WT MEFs stimulated by TGF-β1. Figure 1b shows the functional annotations of 316 genes selected in Figure 1a by gene ontology (GO) analysis. Figure 1c shows a combined diagram of the genes upregulated by the MEF RNA seq. results of Figure 1a and the genes present in each criterion in GSE92592. Figure 1d shows the functional annotations of 37 genes selected in Figure 1c by PANTHER protein class analysis, and Figure 1e shows a dot plot using 5 genes selected in Figure 1d. Figure 1f shows the results of a Western blot performed to compare physical hardness enhancement and the expression of MTHFD2 and α-SMA following TGF-β1 treatment. (MRC5 cells were seeded under soft (0.5 kPa) and hard (glass) conditions and treated with TGF-β1 (4 ng/ml) for 24 hours.) Figure 1g shows the 2D visualization (UMAP dimensionality reduction) results of the pooled data of GSE153935 and a heatmap of the Log2(tumor/normal) level of GSE153935. Specifically, it is the result of clustering according to cell type or diagnosis based on the GEO dataset identified by single cell RNA-sequencing of lung cancer tissue. Figure 1h shows the results of confirming that the expression of various CAF marker genes, including MTHFD2, is increased compared to normal in cancer-associated fibroblasts (CAF) of a fibroblast cluster. Figure 1i shows the results confirming that the expression of MTHFD2 is increased in prostate tumor stromal cells (GSE34312) and colon tumor stroma (GSE31279) compared to normal. Figure 1j shows the results of confirming that when MTHFD2 expression in the tumor stroma of lung cancer patients was checked, MTHFD2 expression was increased in the tumor stroma of patients with a high fibrosis score. Figure 1k shows the data scatter plot of all patients (n = 20) based on MTHFD2 and α-SMA intensity (shown with Pearson correlation coefficient (r) and statistical significance