Use of polyunsaturated fatty acid in combination with acyl-coa ligase (acyl-coa synthetase, ACLY) inhibitor in preparation of anti-tumor drugs

Abstract

The application provides a medicine for treating an immune checkpoint inhibitor-resistant tumor, comprising a polyunsaturated fatty acid, an ACLY inhibitor and an immune checkpoint inhibitor. The polyunsaturated fatty acid comprises one or more of omega-3 polyunsaturated fatty acids and omega-6 polyunsaturated fatty acids. The ACLY inhibitor is selected from one or more of the following: Bempedoic acid, SB 204990, NDI-091143, BMS-303141, small interfering RNA targeting the ACLY gene and short hairpin RNA targeting the ACLY gene. The medicine has good treatment effect on the immune checkpoint inhibitor-resistant tumor.

Description

TECHNICAL FIELD

[0001] The application relates to application of a polyunsaturated fatty acid combined with an ACLY inhibitor to preparation of an antitumor drug and belongs to the technical field of tumor treatment. BACKGROUND

[0002] In recent years, immune checkpoint inhibitors have made significant progress in improving the survival of patients with advanced lung cancer, melanoma, renal cancer, Hodgkin's lymphoma, etc. Among them, immune checkpoint inhibitors targeting programmed cell death receptor 1 (PD-1) or ligand (PD-L1) are of great concern due to their significant clinical efficacy. Although immune checkpoint inhibitors have achieved obvious therapeutic effects in clinical application, the low effective rate in solid tumors is a very prominent limitation, and only a small part of patients (about 20%) get a sustained response and good long-term prognosis, often accompanied by different degrees of immune-related adverse events. The reason for the low effective rate of immune checkpoint inhibitors in solid tumors is that tumors are resistant to immune checkpoint inhibitors. The resistance of tumors to immune checkpoint inhibitors is related to many factors, such as the patient's own characteristics (such as gender / HLA typing), the characteristics of tumor stroma, tumor environmental factors (such as the patient's own microbiome), etc., but the most important of which is the factor of tumor cells themselves. The factor of tumor cells themselves refers to the ability of tumor cells to resist immunotherapy determined by the properties of the tumor cell genome, transcriptome, etc. Due to the complex mechanism of tumor resistance to immune checkpoint inhibitors and numerous influencing factors, how to overcome the resistance of tumors to immune checkpoint inhibitors has become a serious challenge for those skilled in the art. SUMMARY

[0003] The application provides application of a polyunsaturated fatty acid combined with an ACLY inhibitor to preparation of an antitumor drug, which can effectively solve the above problems.

[0004] The application is implemented as follows:

[0005] A drug for treating tumors resistant to immune checkpoint inhibitors, comprising a polyunsaturated fatty acid, an ACLY inhibitor and an immune checkpoint inhibitor.

[0006] As a further improvement, the polyunsaturated fatty acid comprises one or more of omega-3 polyunsaturated fatty acids and omega-6 polyunsaturated fatty acids.

[0007] As a further improvement, the omega-3 polyunsaturated fatty acid is selected from one or more of alpha-linolenic acid, docosahexaenoic acid or eicosapentaenoic acid.

[0008] As a further improvement, the omega-6 polyunsaturated fatty acid is selected from one or more of linoleic acid, gamma-linolenic acid or arachidonic acid.

[0009] As a further improvement, the ACLY inhibitor is selected from one or several of Bempedoic acid, SB 204990, NDI-091143, BMS-303141, small interfering RNA targeting the ACLY gene, short hairpin RNA targeting the ACLY gene.

[0010] As a further improvement, the ACLY inhibitor is selected from Bempedoic acid or short hairpin RNA targeting the ACLY gene.

[0011] As a further improvement, the immune checkpoint inhibitor is one or several of an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, an anti-LAG-3 antibody, an anti-TIGIT antibody, or an anti-TIM-3 antibody.

[0012] Use of a polyunsaturated fatty acid and an ACLY inhibitor in the manufacture of an immune checkpoint inhibitor antitumor sensitizer.

[0013] As a further improvement, the polyunsaturated fatty acid comprises one or several of an omega-3 polyunsaturated fatty acid, an omega-6 polyunsaturated fatty acid.

[0014] As a further improvement, the ACLY inhibitor is selected from one or several of Bempedoic acid, SB 204990, NDI-091143, BMS-303141, small interfering RNA targeting the ACLY gene, short hairpin RNA targeting the ACLY gene.

[0015] As a further improvement, the immune checkpoint inhibitor is an anti-PD-1 antibody or an anti-PD-L1 antibody.

[0016] As a further improvement, the tumor is one or several of a liver cancer, a kidney cancer, a pancreatic cancer, a melanoma, a breast cancer.

[0017] The beneficial effects of the present invention are:

[0018] The medicament for treating immune checkpoint inhibitor-resistant tumors of the present invention comprises a polyunsaturated fatty acid, an ACLY inhibitor, and an immune checkpoint inhibitor, the polyunsaturated fatty acid and the ACLY inhibitor can significantly increase the sensitivity of the immune checkpoint inhibitor to the tumor, and increase the treatment effect of the immune checkpoint inhibitor on the immune checkpoint inhibitor-resistant tumor. BRIEF DESCRIPTION OF DRAWINGS

[0019] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following will briefly introduce the drawings needed to be used in the embodiments. It should be understood that the following drawings only show some of the embodiments of the present application, and therefore should not be regarded as a limitation to the scope. Other related drawings can also be obtained by those of ordinary skill in the art without creative labor.

[0020] Figure 1 The anti-tumor effect diagram provided for the embodiments of the present application.

[0021] Figure 2 The anti-tumor physical diagram provided for the embodiments of the present application. DETAILED DESCRIPTION

[0022] In order to make the purpose, technical solutions and advantages of the embodiments of the present application clearer, the following will combine the drawings in the embodiments of the present application to make a clear and complete description of the technical solutions in the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative labor are within the scope of protection of the present application. Therefore, the following detailed description of the embodiments of the present application provided in the drawings is not intended to limit the scope of the claimed present application, but only represents selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative labor are within the scope of protection of the present application.

[0023] A medicament for treating tumors resistant to immune checkpoint inhibitors, comprising a polyunsaturated fatty acid, an ACLY inhibitor, and an immune checkpoint inhibitor.

[0024] Polyunsaturated fatty acids (PUFA) refer to straight-chain fatty acids containing two or more double bonds and having a carbon chain length of 18-22 carbon atoms. They are usually divided into omega-3 and omega-6 polyunsaturated fatty acids. Polyunsaturated fatty acids can regulate various functions of the human body and also help to remove excess "garbage" in the body. If the human body lacks unsaturated fatty acids, it will affect the physiological functions of the human immune, cardiovascular, reproductive, and endocrine systems, thereby triggering fatal diseases such as high blood pressure, thrombosis, atherosclerosis, rheumatism, and diabetes. Existing studies have shown that adding polyunsaturated fatty acids in the diet also shows potential anti-tumor function.

[0025] ATP citrate lyase (ACLY) is a cytosolic homotetrameric enzyme that catalyzes the conversion of citrate and coenzyme A (CoA) to acetyl-CoA and oxaloacetate, with the concomitant hydrolysis of ATP to ADP and phosphate. ACLY is a key enzyme that links glycolysis and lipid metabolism. In tumor cells characterized by alterations in high-energy metabolism, increased glucose uptake and accelerated glycolytic flux result in increased mitochondrial production of citrate. Once transported into the cytoplasm, citrate is converted by ACLY to acetyl-CoA, a biosynthetic precursor essential for fatty acid synthesis and the mevalonate pathway. Studies have demonstrated that the expression and activity of ACLY are aberrantly expressed in many types of tumors, and pharmacological or genetic inhibition of ACLY significantly inhibits cancer cell proliferation and induces apoptosis.

[0026] In the embodiments of the present application, the combination of polyunsaturated fatty acids and ACLY inhibitors plays a synergistic role, and together improves the sensitivity of immune checkpoint inhibitors to resistant tumors.

[0027] As a further improvement, the polyunsaturated fatty acids include one or more of omega-3 polyunsaturated fatty acids, omega-6 polyunsaturated fatty acids.

[0028] As a further improvement, the omega-3 polyunsaturated fatty acids are selected from one or more of alpha-linolenic acid, docosahexaenoic acid, or eicosapentaenoic acid, but are not limited thereto.

[0029] As a further improvement, the omega-6 polyunsaturated fatty acids are selected from one or more of linoleic acid, gamma-linolenic acid, or arachidonic acid, but are not limited thereto.

[0030] As a further improvement, the ACLY inhibitors are selected from one or more of Bempedoac, SB 204990, NDI-091143, BMS-303141, small interfering RNA targeting ACLY gene, short hairpin RNA targeting ACLY gene, but are not limited thereto.

[0031] As a further improvement, the ACLY inhibitors are selected from Bempedoac or short hairpin RNA targeting ACLY gene.

[0032] As a further improvement, the immune checkpoint inhibitors are one or more of anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-CTLA-4 antibodies, anti-LAG-3 antibodies, anti-TIGIT antibodies, or anti-TIM-3 antibodies, but are not limited thereto.

[0033] As a further improvement, the anti-PD-1 antibodies are nivolumab (Opdivo) or pembrolizumab (Keytruda), and the anti-PD-L1 antibodies are atezolizumab or durvalumab, but are not limited thereto.

[0034] As a further improvement, the ratio of the polyunsaturated fatty acid, the ACLY inhibitor and the immune checkpoint inhibitor needs to be adjusted adaptively according to the kind of the polyunsaturated fatty acid, the ACLY inhibitor and the immune checkpoint inhibitor.

[0035] Use of a polyunsaturated fatty acid and an ACLY inhibitor in the preparation of an immune checkpoint inhibitor antitumor sensitizer.

[0036] As a further improvement, the polyunsaturated fatty acid comprises one or several of omega-3 polyunsaturated fatty acid, omega-6 polyunsaturated fatty acid.

[0037] As a further improvement, the ACLY inhibitor is selected from one or several of Bempedoac, SB 204990, NDI-091143, BMS-303141, small interfering RNA targeting ACLY gene, short hairpin RNA targeting ACLY gene, but not limited thereto.

[0038] As a further improvement, the immune checkpoint inhibitor is one or several of anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, anti-LAG-3 antibody, anti-TIGIT antibody or anti-TIM-3 antibody, but not limited thereto.

[0039] As a further improvement, the tumor is one or several of liver cancer, kidney cancer, pancreatic cancer, melanoma, breast cancer, but not limited thereto.

[0040] Example 1

[0041] Selecting mouse pancreatic cancer cells (Pan02) to verify the immune checkpoint inhibitor treatment of pancreatic cancer animal models resistant to treatment

[0042] Experimental method: mouse pancreatic cancer cells (Pan02) (2x10 6 were inoculated subcutaneously in the forelimbs of 4-6 week old male C57BL / 6 mice, and one week later, the mice were randomly divided into 2 groups: control group and anti-PD-L1 antibody treatment group, 5 mice in each group.

[0043] Anti-PD-L1 antibody treatment group: intraperitoneal injection of anti-PD-L1 antibody atelizumab twice a week (100 μg per mouse).

[0044] Tumor volume was measured every 3 days with a vernier caliper, and the mice were sacrificed on the last day to weigh the tumor.

[0045] The formula for calculating the tumor volume is: tumor volume = 1 / 2 x long diameter of tumor x short diameter of tumor 2 .

[0046] The experimental results are shown in Table 1 andFigure 1 As shown.

[0047] Table 1

[0048]

[0049] Table 1

[0050] like Figure 1 As shown in (A and B), anti-PD-L1 antibody treatment did not have a significant inhibitory effect on the tumor model constructed from mouse pancreatic cancer cells (Pan02), indicating that the model is a tumor model resistant to immune checkpoint therapy.

[0051] Example 2

[0052] ω-3 polyunsaturated fatty acids (eicosapentaenoic acid) combined with short hairpin RNA (shRNA) (sequence: 5′GCTTCATCTCTGGTCTATT-3′ (SEQ ID No. 1)) target and inhibit ACLY, thereby enhancing the sensitivity of pancreatic cancer to anti-PD-L1 antibody therapy.

[0053] Experimental methods: Mouse pancreatic cancer cells (Pan02)((2×10)) were used to treat the cancer cells. 6 (5 mice per mouse) were injected subcutaneously into the forelimbs of 4-6 week old male C57BL / 6 mice to establish an animal model of pancreatic cancer resistant to immune checkpoint inhibitor treatment. One week later, the mice were randomly divided into 4 groups: anti-PD-L1 antibody treatment group, short hairpin RNA (shRNA) targeted inhibition ACLY + anti-PD-L1 antibody treatment group, eicosapentaenoic acid diet + anti-PD-L1 antibody treatment group, and eicosapentaenoic acid diet + short hairpin RNA (shRNA) targeted inhibition ACLY + anti-PD-L1 antibody treatment group, with 5 mice in each group.

[0054] Anti-PD-L1 antibody treatment group: Intraperitoneal injection of atezolizumab (100μg / animal) twice a week.

[0055] Short hairpin RNA (shRNA) targeted inhibition of ACLY + anti-PD-L1 antibody treatment group: Short hairpin RNA (shRNA) was used to target and inhibit the expression of ACLY gene in pancreatic cancer cells (Pan02); the anti-PD-L1 antibody atezolizumab was injected intraperitoneally twice a week (100 μg / animal).

[0056] Eicosapentaenoic acid (EPA) diet + anti-PD-L1 antibody treatment group: EPA was administered by gavage once a day (100 mg / kg); anti-PD-L1 antibody atezolizumab was injected intraperitoneally twice a week (100 μg / animal).

[0057] EPA diet + short hairpin RNA (shRNA) targeting inhibition of ACLY + anti-PD-L1 antibody treatment group: once a day (100 mg / kg) by gavage EPA; twice a week (100 μg / each) intraperitoneal injection of anti-PD-L1 antibody atezolizumab; short hairpin RNA (shRNA) targeting inhibition of ACLY gene expression of pancreatic cancer cells (Pan02).

[0058] Tumor volume was measured every 3 days with a vernier caliper, and the mice were sacrificed on the last day to weigh the tumor.

[0059] The formula for calculating the tumor volume is: tumor volume = 1 / 2 x long diameter of tumor x short diameter 2 .

[0060] Experimental results: as shown in Table 2

[0061] Table 2

[0062]

[0063] As Figure 1 (C-F) shows that after two weeks of group treatment, the tumor volume and weight of the EPA diet + short hairpin RNA (shRNA) targeting inhibition of ACLY + anti-PD-L1 antibody treatment three intervention methods combined group are significantly smaller than the anti-PD-L1 antibody treatment group, the short hairpin RNA (shRNA) targeting inhibition of ACLY + anti-PD-L1 antibody treatment two way combination group, the EPA diet + anti-PD-L1 antibody treatment two way combination group, indicating that polyunsaturated fatty acids (eicosapentaenoic acid) combined with short hairpin RNA (shRNA) targeting inhibition of ACLY enhances the sensitivity of immunotherapy-resistant pancreatic cancer to anti-PD-L1 antibody treatment.

[0064] Wherein, the sum of the tumor inhibition rates of the short hairpin RNA (shRNA) targeting inhibition of ACLY + anti-PD-L1 group and the eicosapentaenoic acid diet + anti-PD-L1 antibody group is less than the tumor inhibition rate of the eicosapentaenoic acid diet + short hairpin RNA (shRNA) targeting inhibition of ACLY + anti-PD-L1 antibody group, indicating that short hairpin RNA (shRNA) and eicosapentaenoic acid play a synergistic effect, and together improve the sensitivity of anti-PD-L1 antibody to resistant tumors.

[0065] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. For those skilled in the art, the present application can have various modifications and changes. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. A medicament for treating tumors resistant to immune checkpoint inhibitors, characterized in that, It includes polyunsaturated fatty acids, ACLY inhibitors, and immune checkpoint inhibitors; the polyunsaturated fatty acid is eicosapentaenoic acid, the ACLY inhibitor is a short hairpin RNA targeting the ACLY gene, and the immune checkpoint inhibitor is atezolizumab.

2. The application of a polyunsaturated fatty acid and an ACLY inhibitor in the preparation of an immune checkpoint inhibitor antitumor sensitizer, characterized in that, The unsaturated fatty acid is eicosapentaenoic acid, the ACLY inhibitor is a short hairpin RNA targeting the ACLY gene, and the immune checkpoint inhibitor is atezolizumab.

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