Development of novel glycolytic enzyme targeting triazines for the treatment of type Ⅱ diabetes and cancer

Abstract

Beginning with the Warburg effect of explaining increased glycolysis in cancer cells, various attempts have been made to understand glucose metabolism in various disease studies. Diabetes, a representative metabolic disease, causes various complications due to abnormal energy metabolism and is known to be highly associated with cancer. In particular, many studies have been conducted on the relationship between hyperinsulinemia and cancer, and many researches on the high correlation between them have been published. Advances in medicine and science have increased life expectancy, but at the same time, the degree of exposure to diseases caused by various factors, such as changes in diet and lifestyle, has also increased. In particular, there is a high increase in the incidence of cancer, including metabolic diseases such as diabetes, hypertension, heart disease. This thesis focused on targeting glycolysis enzymes to develop drugs treating diabetes and cancer. Previous studies have shown that ENOblock targeting the glycolytic enzyme, enolase, increases intracellular glucose uptake, and GAPDS targeting the glycolytic enzyme, glyceraldehyde 3-phosphate dehydrogenase(GAPDH), has anticancer effects. In Part I, as a continuous study of previous studies on the antidiabetic effects of ENOblock in cells and zebrafish, I focused on verifying the antidiabetic effect of ENOblock in db/db mice, a type 2 diabetic mouse model. For 7 weeks, ENOblock was administrated by the intraperitoneal injection, and blood glucose level was monitored during the administration period to check the anti-hyperglycemic effect of ENOblock. After 7 weeks of treatment, blood, liver, heart, kidneys, and adipose tissues were harvested and analyzed. All results were compared with the positive control, Rosiglitazone which has excellent blood sugar control ability but has been withdrawn from the market due to cardiovascular risk, and the potential for side effects of ENOblock was assessed by comparing the side effects known to be caused by rosiglitazone in the ENOblock treatment group. It was confirmed that ENOblock could control blood glucose levels in vivo with 7 weeks of ENOblock treatment. Also, it was confirmed that ENOblock could regulate blood glucose by inhibiting hepatic gluconeogenesis in primary mouse hepatocytes. It was confirmed that ENOblock may have beneficial effects on various complications such as cirrhosis, fatty liver, and heart disease caused by diabetes. In particular, the excellent anti-fibrotic effect demonstrated by ENOblock is considered to be the basis for not only ENOblock but also enolase as a good target in various diseases. In Part II, I established a in vitro tumor microenvironment system to study cross-talk between cancer cells, cancer-associated fibroblasts(CAFs), and tumor-associated macrophages(TAMs) in the tumor microenvironment. Using YD-10B oral cancer conditioned media(CM) and hTERT-fibroblasts(immortalized human gingival fibroblasts), I established a stable system which was enhanced characteristics of inflammatory CAF and confirmed that this system induces monocyte differentiation to macrophage. These results are consistent with our previous laboratory results studied using primary CAF in co-culture system. Also, it was confirmed that GAPDS, which binds GAPDH and inhibit its glycolytic function, suppresses inflammatory CAF characteristics of hTERT-fibroblast induced by YD-10B CM and inhibit macrophage differentiation by targeting hTERT-fibroblasts. Also, GAPDS inhibits macrophage polarization to M2 type. In this thesis, I have shown the anti-diabetic effect of ENOblock and the anti-cancer effect of GAPDS, triazine-based small molecules. ENOblock reduced a hyperglycemia in type 2 diabetes, and improved symptoms of various complications, such as hepatic steatosis, fibrosis, cardiac hypertrophy, and fibrosis. I established a stable in vitro system to study cross-talk between CAF and TAM in the tumor microenvironment. In this system, GAPDS inhibited macrophage differentiation induced by cancer CM-stimulated hTERT-fibroblast. Taken together, I suggest enolase as a new target and ENOblock as a lead compound for diabetes treatment and suggest GAPDH as a new target and GAPDS as a lead compound for cancer treatment.