Thiogeranyl geranyl pyrophosphate GGSPP and application thereof in preparation of liver cancer drugs

Abstract

The invention discloses thiogeranyl geranyl pyrophosphate GGSPP and application thereof in preparation of liver cancer drugs, the structural formula of the GGSPP is as shown in formula (I): # imgabs0 # GGSPP specifically binds and activates human fructose-1, 6-diphosphatase 1FBP1 protein, up-regulates the enzymatic activity of FBP1, and reverses the Warburg effect of tumor cells, thereby inhibiting proliferation, migration and development of hepatocellular carcinoma. The invention not only clarifies the key effect of the GGSP-FBP1 axis in the occurrence and development of liver cancer, but also provides an efficient, specific and mechanism-clear anti-liver cancer candidate drug, opens up a new direction for the treatment of metabolism-related tumors, and has important scientific significance and clinical application prospects.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and particularly to a thio-geranylgeranyl pyrophosphate (GGSPP) and its application in the preparation of drugs for liver cancer. Background Art

[0002] Disorders of glycolipid metabolism lead to HCC. The regulation of glycolipid metabolic homeostasis is the basis for maintaining the life activities of the body. In China, due to the changes in lifestyle and dietary structure in recent decades, the glycolipid metabolism patterns of some people have changed accordingly, and the incidence of metabolic syndromes such as obesity and diabetes has increased significantly. As the central organ of metabolism, the liver bears the brunt. Long-term disorders of glycolipid metabolism trigger chronic liver diseases such as non-alcoholic fatty liver disease (NAFLD) and liver fibrosis. Recent research results have shown that in addition to viral hepatitis and alcoholic liver disease, metabolic syndromes and chronic liver diseases such as NAFLD are also important inducements for primary liver cancer. At the same time, by controlling the glycolipid metabolism of cancer cells and using various strategies to specifically block the energy sources of cancer cells to achieve cancer prevention and treatment, it has also attracted increasing attention. Therefore, glycolipid metabolic homeostasis plays an important role in the occurrence, development, prevention, early diagnosis and treatment of primary liver cancer.

[0003] Currently, there is a lack of effective clinical treatment drugs for HCC. As the main type of primary liver cancer, hepatocellular carcinoma (HCC) is one of the most common malignant tumors in the body worldwide and one of the most common causes of tumor-related death. In addition to surgical treatment, for patients with advanced liver cancer, the multikinase inhibitor Sorafenib is the only drug approved by multiple countries for the systemic treatment of advanced HCC, but it is expensive, has special adverse reactions, and its effectiveness for different patients is also questionable. In addition, in terms of new drug development, clinical trials of other molecular targeted agents such as Sunitinib, Brivanib and Linifanib for the treatment of liver cancer have all obtained a series of negative results. "New uses of old drugs" has become a new trend in drug development in the high-investment and high-risk drug development process because it can greatly shorten the development time, cost and clinical application cycle. As the most common type of lipid-lowering prescription drugs in clinical practice, statins are widely used and are mainly used to treat and prevent hyperlipidemia associated with metabolic syndrome and various cardiovascular diseases. A large amount of recent clinical statistical data shows that statins can reduce the risk of death in patients with various cancers such as HCC, suggesting the possibility of HCC as a new indication for statins. However, there is still uncertainty about the role of statins in the prevention and treatment of HCC. This may be related to the fact that statins inhibit 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) located upstream of the mevalonate pathway, resulting in the obstruction of the synthesis of a variety of metabolic small molecules with important biological functions downstream, such as farnesyl pyrophosphate (FPP), squalene, cholesterol (Cholesterol), geranylgeranyl pyrophosphate (GGPP) and ubiquinone (Ubiquinone) (such as Figure 1 shown).

[0004] The body's response to the loss of small molecules in the mevalonate pathway is extremely complex, which may be one of the important reasons why the therapeutic effects of statins on different cancer types and even different patients vary significantly. Elucidating the role of these small molecules in the mevalonate pathway in the occurrence and development of HCC is of great significance for the application of statins in the treatment of HCC and the further development of related treatment plans. Figure 1It is a mevalonate pathway diagram. Among them, geranylgeranyl pyrophosphate (GGPP) is one of the important metabolic small molecules downstream of the mevalonate pathway and is another important product of farnesyl pyrophosphate (FPP) other than cholesterol (as Figure 1 shown). The main functions of FPP and GGPP that have been reported are to participate in an important protein lipid acylation modification - isoprenylation modification as substrates, and by changing the hydrophobicity of certain specific proteins respectively, their localization and activity in the membrane system are changed, regulating the downstream signaling pathway, and thus playing specific functions.

[0005] The applicant previously discovered the association between GGSPP precursor GGPP and FBP1 and hepatocellular carcinoma. Geranylgeranyl pyrophosphate synthase 1 (GGPPS1) is the synthase in the body responsible for catalyzing the conversion of FPP to GGPP. The applicant used Ggpps1-specific knockout mice in different organ tissues to study the effects of relative changes in FPP and GGPP levels on the body's functions. Liver-specific Ggpps1 knockout mice were more prone to form primary liver cancer during DEN chemical mutagenesis; while the detection of liver cancer samples from HCC patients clinically found that the expression level of GGPP synthase GGPPS1 was positively correlated with the malignancy of HCC, and HCC patients with high GGPPS1 expression had a better prognosis, suggesting that GGPPS1 and GGPP may protect the liver through compensatory upregulation during the progression of HCC. At the same time, the analysis of Clarivate Analytics' Metadrug software also showed that GGPP has significant anti-tumor activity. These results suggest that GGPP plays an important role in the regulation of glucose and lipid metabolism in the liver and the occurrence and development of liver cancer, but the direct target proteins regulated by it and the corresponding mechanisms of action are currently poorly understood.

[0006] FBP1 is a key rate-limiting enzyme in the process of gluconeogenesis and has also been confirmed to be an important tumor suppressor protein. Multiple studies have reported that the deletion, mutation and decreased expression of FBP1 can lead to the occurrence and development of various cancers such as hepatocellular carcinoma, renal clear cell carcinoma, non-small cell lung cancer, breast cancer, etc. In HCC, the deletion or reduction of FBP1 may lead to glucose and lipid metabolism disorders and abnormal lipid accumulation by affecting the Warburg effect, providing a large amount of substances and energy for cancer cells and accelerating the occurrence and development of HCC. And statistical data also show that patients with high FBP1 expression levels in HCC have significantly better prognoses, which makes FBP1 a potential prognostic predictor for HCC. However, no reports on small molecule activators targeting FBP1 have been found so far.

[0007] Currently, there is a lack of an application of a small molecule compound with geranylgeranyl pyrophosphate (GGPP) as a precursor in the preparation of anti-hepatocellular carcinoma drugs by binding to and activating human fructose-1,6-bisphosphatase 1 (FBP1). Summary of the Invention

[0008] To solve the problems of the prior art, the object of the present invention is to provide a thio-geranylgeranyl pyrophosphate (GGSPP) and its application in the preparation of liver cancer drugs.

[0009] To achieve the above object, the present invention adopts the following technical solutions:

[0010] In the first aspect, the present application provides an application of thio-geranylgeranyl pyrophosphate (GGSPP) in the preparation of liver cancer drugs.

[0011] In the second aspect, the present application provides an anti-hepatocellular carcinoma drug composition.

[0012] In the third aspect, the present application provides a method for inhibiting the occurrence and development of hepatocellular carcinoma.

[0013] The first aspect of the present application provides an application of thio-geranylgeranyl pyrophosphate (GGSPP) in the preparation of liver cancer drugs. The structural formula of GGSPP is shown in formula (I):

[0014]

[0015] GGSPP inhibits the proliferation, migration and development of hepatocellular carcinoma by specifically binding to and activating human fructose-1,6-bisphosphatase 1 (FBP1) protein, up-regulating the enzyme activity of FBP1, and reversing the Warburg effect of tumor cells.

[0016] Furthermore, the amino acid sequence of human FBP1 protein is shown in SEQ ID No.1.

[0017] Furthermore, GGSPP promotes gluconeogenesis and inhibits glycolysis by up-regulating the enzyme activity of FBP1, and at the same time coordinates the metabolic balance of glucose synthesis and cholesterol biosynthesis pathways.

[0018] Even further, the inhibitory effect of GGSPP on liver cancer cells is manifested as significantly reducing the proliferation ability and migration ability of HepG2 and Huh7 cell lines.

[0019] Furthermore, GGSPP inhibits the malignant progression of hepatocellular carcinoma by inhibiting the Warburg effect and cholesterol synthesis, reducing the glycolysis level in liver cancer tissues, and improving cholesterol metabolic homeostasis.

[0020] Furthermore, compared with its precursor GGPP, GGSPP has significantly reduced isoprenylation ability and higher binding affinity for FBP1, and the dissociation constant KD is 400 - 600 nM.

[0021] Furthermore, the anti - hepatocellular carcinoma drug is applicable to the treatment of hepatocellular carcinoma caused by glycolipid metabolism disorders or FBP1 expression deficiency.

[0022] The second aspect of the present application provides an anti - hepatocellular carcinoma drug composition, comprising a therapeutically effective amount of the compound GGSPP, and a pharmaceutically acceptable carrier or excipient.

[0023] Furthermore, the dosage form of the drug composition is an oral preparation, an injection or a targeted delivery preparation.

[0024] The third aspect of the present application provides a method for inhibiting the occurrence and development of hepatocellular carcinoma, by administering a therapeutically effective amount of the compound GGSPP to a patient to activate FBP1 and reverse the Warburg effect, thereby inhibiting the proliferation and migration of liver cancer cells.

[0025] Beneficial effects: The present invention not only clarifies the key role of the GGSPP - FBP1 axis in the occurrence and development of liver cancer, but also provides a highly efficient, specific and clearly - mechanism anti - liver cancer candidate drug, opening up a new direction for the treatment of metabolism - related tumors, and having important scientific significance and clinical application prospects.

[0026] Compared with the prior art, the present invention has the following advantages:

[0027] (1) Specific targeting effect: The present invention for the first time reveals that the compound GGSPP directly regulates the balance of liver glycolipid metabolism by specifically binding to and activating human fructose - 1,6 - bisphosphatase 1 (FBP1). Compared with its precursor GGPP, GGSPP significantly reduces the isoprenylation ability through structural modification (thiol modification), avoiding non - specific effects caused by interference of isoprenylation function, and thus more efficiently and specifically targets FBP1 (dissociation constant K D is 400 - 600 nM), improving the precision and safety of the drug.

[0028] (2) Reversing the Warburg effect: GGSPP promotes gluconeogenesis and inhibits glycolysis by up - regulating the enzyme activity of FBP1, effectively reversing the Warburg effect of tumor cells (i.e., the metabolic characteristics of cancer cells relying on glycolysis), cutting off the energy and material supply of liver cancer cells, and significantly inhibiting their proliferation and migration ability (the inhibition rate is better than that of natural GGPP in HepG2 and Huh7 cell models).

[0029] (3) Metabolic balance regulation: GGSPP coordinates the metabolic balance of glucose synthesis and cholesterol biosynthesis pathways by activating FBP1, reducing abnormal glycolysis in liver cancer tissues ( Figure 6 E) and restoring cholesterol metabolic homeostasis (Figure 6 F), inhibiting the malignant progression of hepatocellular carcinoma at the molecular level, providing a new strategy for the treatment of liver cancer related to metabolic disorders.

[0030] (4) Clinical translation potential: Significant efficacy: In a mouse liver cancer model, GGSPP supplementation significantly inhibited tumorigenesis and development ( Figure 6 C-D), and the effect was better than that of existing statins, avoiding the problem of metabolic molecule deficiency caused by the latter's inhibition of the upstream of the mevalonate pathway.

[0031] (5) Target innovation: As a novel tumor suppressor protein, there is currently no report on small molecule activators targeting FBP1. The present invention fills this gap in the field, providing a new target and drug candidate molecule for the treatment of liver cancer.

[0032] (6) Wide application: GGSPP is not only applicable to hepatocellular carcinoma, and its mechanism of regulating glucose and lipid metabolism balance may be extended to other cancer types dependent on the Warburg effect (such as renal cancer, lung cancer, etc.), with potential broad-spectrum anti-tumor value.

[0033] (7) R & D advantages: Based on the structural optimization of GGPP precursors, GGSPP follows the strategy of "repurposing old drugs", which can significantly shorten the drug R & D cycle and cost. At the same time, combined with multi-level experimental verification (molecular, cellular, animal and clinical data), it lays a solid foundation for subsequent clinical trials. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] In order to more clearly illustrate the technical solutions in the test examples of the present application, the following will briefly introduce the drawings required for use in the test examples or the description of the prior art. Obviously, the drawings in the following description are only some test examples of the present application. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0035] ABBREVIATION DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 It is a diagram of the mevalonate pathway.

[0037] Figure 2 It is a verification diagram of BLI and ITC for the binding of GGSPP of the present invention to FBP1 protein. Figure 2 A shows that the ability of GGSPP molecule of the present invention to participate in isoprenylation is significantly reduced compared with GGPP; Figure 2 B is the BLI verification of the binding of GGSPP and FBP1 of the present invention; Figure 2 C is the ITC verification of the binding of GGSPP and FBP1 of the present invention.

[0038] Figure 3This is the figure showing that GGSPP of the present invention significantly upregulates the enzymatic activity of FBP1 after binding to FBP1. Figure 3 A shows that treatment with GGSPP of the present invention significantly upregulates the enzymatic activity of FBP1, and the activation effect is better than that of natural GGPP; Figure 3 B- Figure 3 C shows that treatment with GGSPP of the present invention significantly increases glucose production in primary hepatocytes ( Figure 3 B), but inhibits cholesterol production ( Figure 3 C); Figure 3 D- Figure 3 G shows that ECAR detection of the present invention indicates that treatment with GGSPP inhibits glycolysis in primary hepatocytes ( Figure 3 D), and all indicators of glycolysis decrease ( Figure 3 E- Figure 3 G).

[0039] Figure 4 This is the figure showing that GGSPP of the present invention inhibits the migration and proliferation of HepG2 liver cancer cells. Figure 4 A- Figure 4 B shows that treatment with GGSPP of the present invention significantly inhibits the migration of HepG2 liver cancer cells, and the inhibition effect is better than that of natural GGPP, Figure 4 B is Figure 4 the statistical result of A; Figure 4 C shows that treatment with GGSPP of the present invention significantly inhibits the proliferation of HepG2 liver cancer cells, and the inhibition effect is better than that of natural GGPP.

[0040] Figure 5 This is the figure showing that GGSPP of the present invention inhibits the migration and proliferation of Huh 7 liver cancer cells. Figure 5 A- Figure 5 B shows that treatment with GGSPP of the present invention significantly inhibits the migration of Huh 7 liver cancer cells, and the inhibition effect is better than that of natural GGPP, Figure 5 Figure B is Figure 5 the statistical result of Figure A; Figure 5 C shows that treatment with GGSPP of the present invention significantly inhibits the proliferation of Huh 7 liver cancer cells, and the inhibition effect is better than that of natural GGPP.

[0041] Figure 6 This is the figure showing that GGSPP of the present invention can inhibit the occurrence and development of mouse hepatocellular carcinoma. Figure 6 A is the timeline of establishing a mouse primary hepatocellular carcinoma model by high-fat diet and chemical mutagenesis of the present invention; Figure 6 B is the reversal situation of the normal control group, the high-fat diet and chemically induced mouse primary hepatocellular carcinoma group, and the GGSPP supplementation of the present invention; Figure 6 C- Figure 6D is the statistical analysis of the malignancy comparison, tumor number, and volume of the three groups of mouse hepatocellular carcinoma in B of the present invention; Figure 6 E- Figure 6 F is the detection result of glucose level ( Figure 6 E) and cholesterol level ( Figure 6 F) in the liver cancer tissues of different groups of mice in the liver cancer mouse model of the present invention. Detailed Embodiments

[0042] The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments. The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. The experimental methods in the following examples of the present invention that do not specify specific conditions are generally carried out under conventional conditions or according to the conditions recommended by the manufacturer. All common chemical reagents used in the examples are commercially available products.

[0043] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly understood by those skilled in the technical field to which the present invention belongs. The terms used in the description of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0044] The terms "comprising" and "having" and any variations thereof in the present invention are intended to cover non-exclusive inclusion. For example, a process, method, device, product, or equipment that includes a series of steps is not limited to the listed steps or modules, but may optionally further include steps not listed, or may optionally further include other steps inherent to these processes, methods, products, or equipment.

[0045] As used in the present invention, "a plurality of" means two or more. "And / or" describes the association relationship of associated objects and indicates that three relationships may exist. For example, A and / or B may represent: A exists alone, A and B exist simultaneously, and B exists alone. The character " / " generally represents an "or" relationship between the associated objects before and after.

[0046] In the first aspect of the embodiments of the present application, there is provided an application of thio-geranylgeranyl pyrophosphate GGSPP in the preparation of a liver cancer drug. The structural formula of GGSPP is shown in formula (I):

[0047]

[0048] GGSPP inhibits the proliferation, migration, and development of hepatocellular carcinoma by specifically binding to and activating the human fructose-1,6-bisphosphatase 1 FBP1 protein, upregulating the enzyme activity of FBP1, and reversing the Warburg effect of tumor cells.

[0049] In some embodiments, the amino acid sequence of the human FBP1 protein is as shown in SEQ ID No.1.

[0050] In some embodiments, GGSPP promotes gluconeogenesis and inhibits glycolysis by upregulating the enzymatic activity of FBP1, while coordinating the metabolic balance between glucose synthesis and cholesterol biosynthesis pathways.

[0051] In some embodiments, the inhibitory effect of GGSPP on liver cancer cells is manifested as a significant reduction in the proliferation and migration abilities of the HepG2 and Huh7 cell lines.

[0052] In some embodiments, GGSPP inhibits the malignant progression of hepatocellular carcinoma by inhibiting the Warburg effect and cholesterol synthesis, reducing the glycolysis level in liver cancer tissues, and improving cholesterol metabolic homeostasis.

[0053] In some embodiments, compared with its precursor GGPP, GGSPP has a significantly reduced isoprenylation ability and a higher binding affinity for FBP1, and the dissociation constant K D is 400 nM.

[0054] In some embodiments, compared with its precursor GGPP, GGSPP has a significantly reduced isoprenylation ability and a higher binding affinity for FBP1, and the dissociation constant K D is 500 nM.

[0055] In some embodiments, compared with its precursor GGPP, GGSPP has a significantly reduced isoprenylation ability and a higher binding affinity for FBP1, and the dissociation constant K D is 600 nM.

[0056] In some embodiments, the anti-hepatocellular carcinoma drug is applicable to the treatment of hepatocellular carcinoma caused by glycolipid metabolism disorders or FBP1 expression deficiency.

[0057] In a second aspect of the embodiments of the present application, an anti-hepatocellular carcinoma drug composition is provided, comprising a therapeutically effective amount of the compound GGSPP and a pharmaceutically acceptable carrier or excipient.

[0058] In some embodiments, the dosage form of the drug composition is an oral preparation, an injection or a targeted delivery preparation.

[0059] In a third aspect of the embodiments of the present application, a method for inhibiting the occurrence and development of hepatocellular carcinoma is provided, by administering a therapeutically effective amount of the compound GGSPP to a patient to activate FBP1 and reverse the Warburg effect, thereby inhibiting the proliferation and migration of liver cancer cells.

[0060] Example 1

[0061] Use of a thio-geranylgeranyl pyrophosphate GGSPP in the preparation of a hepatocellular carcinoma drug, wherein the structural formula of GGSPP is as shown in formula (I):

[0062]

[0063] GGSPP inhibits the proliferation, migration and development of hepatocellular carcinoma by specifically binding to and activating the human fructose-1,6-bisphosphatase 1 FBP1 protein, upregulating the enzyme activity of FBP1, and reversing the Warburg effect of tumor cells.

[0064] The amino acid sequence of the human FBP1 protein is as shown in SEQ ID No.1.

[0065] GGSPP promotes gluconeogenesis and inhibits glycolysis by upregulating the enzyme activity of FBP1, and at the same time coordinates the metabolic balance of the glucose synthesis and cholesterol biosynthesis pathways.

[0066] The inhibitory effect of GGSPP on hepatocellular carcinoma cells is manifested as a significant reduction in the proliferation and migration abilities of the HepG2 and Huh7 cell lines.

[0067] GGSPP inhibits the malignant progression of hepatocellular carcinoma by inhibiting the Warburg effect and cholesterol synthesis, reducing the glycolysis level in hepatocellular carcinoma tissues, and improving cholesterol metabolic homeostasis.

[0068] Compared with its precursor GGPP, GGSPP has significantly reduced isoprenylation ability and higher binding affinity for FBP1, and the dissociation constant K D is 600 nM.

[0069] The anti-hepatocellular carcinoma drug is applicable to the treatment of hepatocellular carcinoma caused by glycolipid metabolism disorders or FBP1 expression deficiency.

[0070] Example 2

[0071] The difference between Example 2 and Example 1 is that the dissociation constant K D is 400 nM.

[0072] Example 3

[0073] The difference between Example 2 and Example 1 is that the dissociation constant K D is 500 nM.

[0074] Example 4

[0075] An anti-hepatocellular carcinoma drug composition of the present invention comprises a therapeutically effective amount of a compound GGSPP and a pharmaceutically acceptable carrier or excipient. The dosage form of the drug composition is an injection.

[0076] Example 5

[0077] Example 5 is different from Example 4 in that the dosage form of the pharmaceutical composition is a targeted delivery preparation.

[0078] Example 6

[0079] Example 6 is different from Example 4 in that the dosage form of the pharmaceutical composition is an oral preparation.

[0080] Example 7

[0081] A method for inhibiting the occurrence and development of hepatocellular carcinoma according to the present invention is to administer a therapeutically effective amount of the compound GGSPP to a patient to activate FBP1 and reverse the Warburg effect, thereby inhibiting the proliferation and migration of liver cancer cells.

[0082] Test Example 1

[0083] GGSPP binds to and activates FBP1

[0084] 1. Verification of specific binding of GGSPP to FBP1 by BLI and ITC

[0085] Previous studies by the applicant have shown that FBP1 is a potential candidate target protein for GGPP. Based on this discovery, GGSPP was synthesized according to the literature reports. First, the ability of GGPP and GGSPP to induce isoprenylation was compared, and the results showed that the isoprenylation ability of GGSPP was significantly reduced ( Figure 2 A), and it can bind to FBP1 more specifically and efficiently. Then, various methods were further used to verify the binding of GGSPP and FBP1. Experiments such as biomembrane interference technology (BLI) and isothermal titration calorimetry (ITC) showed that the recombinant FBP1 protein purified from Escherichia coli could specifically bind to GGSPP in vitro. The dissociation constant K D value of the binding of FBP1 and GGSPP detected by BLI was 510 nM ( Figure 2 B), and the K D value detected by ITC was 493.5 nM ( Figure 2 C). Figure 2 This is the BLI and ITC verification diagram of the binding of GGSPP of the present invention to FBP1 protein. Figure 2 A shows that the ability of the GGSPP molecule of the present invention to participate in isoprenylation is significantly reduced compared to GGPP; Figure 2 B is the BLI verification of the binding of GGSPP and FBP1 of the present invention; Figure 2 C is the ITC verification of the binding of GGSPP and FBP1 of the present invention.

[0086] 2. GGSPP binding upregulates FBP1 enzyme activity and promotes gluconeogenesis, reverses the Warburg effect of tumors, and inhibits glycolysis and cholesterol synthesis; Figure 3After GGSPP of the present invention binds to FBP1, the enzymatic activity of FBP1 is significantly upregulated; Figure 3 A. Treatment with GGSPP of the present invention significantly upregulates the enzymatic activity of FBP1, and the activation effect is better than that of natural GGPP; Figure 3 B- Figure 3 C. Treatment with GGSPP of the present invention significantly increases glucose production in primary hepatocytes ( Figure 3 B), but inhibits cholesterol production ( Figure 3 C); Figure 3 D- Figure 3 G. ECAR detection of the present invention shows that treatment with GGSPP inhibits glycolysis in primary hepatocytes ( Figure 3 D), and all indicators of glycolysis decrease ( Figure 3 E- Figure 3 G).

[0087] Experimental Example 2

[0088] GGSPP binds to and activates FBP1 to inhibit the occurrence and development of hepatocellular carcinoma

[0089] In view of the important regulatory role of GGSPP in the activity of FBP1 and the important function of FBP1 itself as a tumor suppressor protein, the present invention first confirmed in the hepatoma cell lines HepG2 and Huh7 that treatment with GGSPP can significantly inhibit the migration and proliferation of hepatoma cells ( Figure 4 , Figure 5 ). Figure 4 This is a diagram showing that GGSPP of the present invention inhibits the migration and proliferation of HepG2 hepatoma cells. Figure 4 A- Figure 4 B. Treatment with GGSPP of the present invention significantly inhibits the migration of HepG2 hepatoma cells, and the inhibition effect is better than that of natural GGPP. Figure 4 B is Figure 4 the statistical result of A of the present invention; Figure 4 C. Treatment with GGSPP of the present invention significantly inhibits the proliferation of HepG2 hepatoma cells, and the inhibition effect is better than that of natural GGPP. Figure 5 This is a diagram showing that GGSPP of the present invention inhibits the migration and proliferation of Huh7 hepatoma cells. Figure 5 A- Figure 5 B. Treatment with GGSPP of the present invention significantly inhibits the migration of Huh7 hepatoma cells, and the inhibition effect is better than that of natural GGPP. Figure 5 B is Figure 5 the statistical result of A; Figure 5 C. Treatment with GGSPP of the present invention significantly inhibits the proliferation of Huh7 hepatoma cells, and the inhibition effect is better than that of natural GGPP.

[0090] This invention studied the role of the combination of GGSPP and FBP1 in inhibiting the occurrence and development of mouse hepatocellular carcinoma. The results showed that when using a high-fat diet and chemical mutagenesis to create a mouse model of primary hepatocellular carcinoma, wild-type mice developed liver cancer ( Figure 6 A- Figure 6 B); while replenishing GGSPP to the mice with liver cancer model could significantly inhibit the occurrence and development of mouse liver cancer ( Figure 6 C- Figure 6 D), and this inhibitory effect was related to GGSPP promoting gluconeogenesis, inhibiting the Warburg effect, and cholesterol synthesis ( Figure 6 E). Figure 6 Figure for GGSPP of this invention can couple glucose metabolism and cholesterol synthesis to inhibit the occurrence and development of mouse hepatocellular carcinoma; Figure 6 A is the timeline of creating a mouse model of primary hepatocellular carcinoma with a high-fat diet and chemical mutagenesis of this invention; Figure 6 B is the reversal situation of the normal control group, the high-fat diet and chemically induced mouse primary hepatocellular carcinoma group, and the replenishment of GGSPP of this invention; Figure 6 C- Figure 6 D is the statistical analysis of the malignancy degree comparison, tumor number, and volume size of hepatocellular carcinoma in the three groups of mice in B of this invention; Figure 6 E- Figure 6 F is the glucose level ( Figure 6 E) and cholesterol level ( Figure 6 F) detection results in the liver cancer tissues of different groups of mice in the liver cancer mouse model of this invention.

[0091] The above shows and describes the basic principles, main features, and advantages of this invention. Those skilled in the art should understand that this invention is not limited by the above test examples. What is described in the above test examples and the specification only illustrates the principles of this invention. Without departing from the spirit and scope of this invention, this invention will have various changes and improvements. The scope of protection required by this invention is defined by the appended claims, the specification, and their equivalents.

Claims

1. An application of thiogeranylgeranyl pyrophosphate (GGSPP) in the preparation of liver cancer drugs, characterized in that: The structural formula of the GGSPP is shown in formula (I): GGSPP specifically binds to and activates human fructose-1,6-bisphosphatase 1 FBP1 protein, upregulates the enzymatic activity of FBP1, reverses the Warburg effect of tumor cells, and thus inhibits the proliferation, migration and development of hepatocellular carcinoma.

2. The use according to claim 1, characterized in that: The amino acid sequence of the human FBP1 protein is shown in SEQ ID No.

1.

3. The use according to claim 1, characterized in that: The GGSPP promotes gluconeogenesis and inhibits glycolysis by upregulating the enzymatic activity of FBP1, while coordinating the metabolic balance of glucose synthesis and cholesterol biosynthesis pathways.

4. The use according to claim 1, characterized in that: The inhibitory effect of GGSPP on liver cancer cells is manifested in significantly reducing the proliferation and migration abilities of HepG2 and Huh7 cell lines.

5. The use according to claim 1, characterized in that: The GGSPP inhibits the Warburg effect and cholesterol synthesis, reduces the level of glycolysis in liver cancer tissue, and improves cholesterol metabolism homeostasis, thereby inhibiting the malignant progression of hepatocellular carcinoma.

6. The use according to claim 1, characterized in that: Compared with its precursor GGPP, the GGSPP has significantly lower isoprenylation ability and higher binding affinity with FBP1, with a dissociation constant K D It is 400-600nM.

7. The use according to claim 1, characterized in that: The anti-hepatocellular carcinoma drug is suitable for treating hepatocellular carcinoma caused by glucose and lipid metabolism disorder or FBP1 expression deficiency.

8. An anti-hepatocellular carcinoma pharmaceutical composition, characterized in that: It comprises a therapeutically effective amount of the compound GGSPP according to claim 1, and a pharmaceutically acceptable carrier or excipient.

9. The pharmaceutical composition according to claim 8, characterized in that: The dosage form of the pharmaceutical composition is an oral preparation, an injection or a targeted delivery preparation.

10. A method for inhibiting the occurrence and development of hepatocellular carcinoma, characterized in that: A therapeutically effective amount of the compound GGSPP according to claim 1 is administered to a patient to activate FBP1 and reverse the Warburg effect, thereby inhibiting the proliferation and migration of liver cancer cells.

Images