Application of KAT6A inhibitor combined with PD-1 monoclonal antibody in preparation of medicine for treating colorectal cancer

Abstract

The invention discloses application of a KAT6A inhibitor combined with a PD-1 monoclonal antibody in preparation of a medicine for treating colorectal cancer, relates to the technical field of tumor treatment, and aims to solve the problems that an existing immune checkpoint inhibitor is poor in MSS type intestinal cancer treatment effect and has drug resistance. The invention provides application of a KAT6A inhibitor combined with a PD-1 monoclonal antibody in preparation of a medicine for treating colorectal cancer and application of the KAT6A inhibitor in preparation of the medicine for treating the colorectal cancer. The invention also provides a pharmaceutical composition for resisting colorectal cancer. The pharmaceutical composition comprises the KAT6A inhibitor and the PD-1 monoclonal antibody. The KAT6A inhibitor is combined for use, so that the immunotherapy effect of the PD-1 monoclonal antibody is remarkably improved, and the growth of subcutaneous transplanted tumors of mice is inhibited. The traditional Chinese medicine is used for treating colorectal cancer.

Description

Technical Field

[0001] The present invention relates to the technical field of tumor treatment, and specifically relates to the application of a KAT6A inhibitor combined with a PD-1 monoclonal antibody in the preparation of a drug for treating colorectal cancer. Background Art

[0002] Colorectal cancer (CRC) is one of the most common malignant tumors globally, with its incidence ranking third among various cancers and its mortality rate ranking second. Currently, significant progress has been made in the immunotherapy of colorectal cancer, and immune checkpoint inhibitors (ICIs) have been approved for the treatment of advanced colorectal cancer with high microsatellite instability (MSI-H) or defective mismatch repair (dMMR).

[0003] Microsatellite stable (MSS) colorectal cancer accounts for approximately 95% of advanced colorectal cancer, and 85%-95% of patients with MSS colorectal cancer are initially resistant to single-agent ICI, with very limited treatment effects and usually a low objective response rate (<5%). Patients with MSI-H colorectal cancer are relatively sensitive to ICI treatment. Even so, a considerable number of patients with MSI-H colorectal cancer are initially resistant (ineffective in initial treatment) or secondarily resistant (initially effective but with subsequent disease progression) to ICI. Therefore, there is an urgent need to find more effective immunotherapy combination strategies for the treatment of CRC. Summary of the Invention

[0004] The present invention aims to solve the problems of poor treatment effects and drug resistance of existing immune checkpoint inhibitors in the treatment of MSS colorectal cancer, and provides the application of a KAT6A inhibitor combined with a PD-1 monoclonal antibody in the preparation of a drug for treating colorectal cancer.

[0005] The present invention provides the application of a KAT6A inhibitor combined with a PD-1 monoclonal antibody in the preparation of a drug for treating colorectal cancer.

[0006] Further, the KAT6A inhibitor is WM-1119 (MCE, HY-102058).

[0007] Further, the colorectal cancer is MSS colorectal cancer and MSI-H colorectal cancer.

[0008] Further, the PD-1 monoclonal antibody is CD279.

[0009] The present invention provides the application of a KAT6A inhibitor in the preparation of a drug for treating colorectal cancer.

[0010] Further, the colorectal cancer is MSS colorectal cancer and MSI-H colorectal cancer.

[0011] The present invention also provides an anti-colorectal cancer drug composition, which comprises a KAT6A inhibitor and a PD-1 monoclonal antibody.

[0012] Advantages of the present invention:

[0013] The present invention provides the application of the KAT6A gene target in anti-tumor treatment, and proposes a dual therapy regimen combining a KAT6A inhibitor and a PD-1 monoclonal antibody. In a preclinical animal model, compared with the control group, intraperitoneal injection of the KAT6A inhibitor alone can inhibit the growth of colorectal cancer tumors. At the same time, the combined regimen of the KAT6A inhibitor and immune therapy also has obvious therapeutic effects. The combined use of the KAT6A inhibitor significantly enhances the effect of PD-1 monoclonal antibody immunotherapy and inhibits the growth of subcutaneous transplanted tumors in mice.

[0014] KAT6A knockdown activates the cGAS / STING signaling pathway, thereby activating the interferon (IFN) response, promoting CD8+ T cell infiltration, and sensitizing colorectal cancer immunotherapy. It increases the sensitivity of MSS-type murine colorectal cancer to PD-1 monoclonal antibody treatment and reverses the resistance of MSS-type colorectal cancer to immunotherapy.

[0015] The present invention first proves that KAT6A may be a potential molecular marker and an important therapeutic target for the failure of immunotherapy for MSS colorectal cancer, and at the same time proves that the KAT6A inhibitor sensitizes immunotherapy. It provides a new marker for the immunotherapy of colorectal cancer. Description of the Drawings

[0016] Figure 1 It is a flow chart of grouping and administering drugs to mice with MSS-type colorectal cancer model;

[0017] Figure 2 It is a comparison of the volumes of subcutaneous transplanted tumors in mice homologous to CT26 cells in different treatment groups;

[0018] Figure 3 It is a comparison of the weights of subcutaneous transplanted tumors in mice homologous to CT26 cells in different treatment groups;

[0019] Figure 4 It is the change of the volume of subcutaneous transplanted tumors in mice homologous to CT26 cells in different treatment groups over the administration time;

[0020] Figure 5 It is a flow chart of grouping and administering drugs to mice with MSI-H type colorectal cancer model;

[0021] Figure 6 It is a comparison of the volumes of subcutaneous transplanted tumors in mice homologous to MC38 cells in different treatment groups;

[0022] Figure 7 It is a comparison of the weights of subcutaneous transplanted tumors in mice homologous to MC38 cells in different treatment groups;

[0023] Figure 8The change in the volume of MC38 cell syngeneic subcutaneous xenografts in mice of different treatment groups over the administration time;

[0024] Figure 9 The waterfall plot for comparing the efficacy of immune checkpoint inhibitor (ICI) treatment in colorectal cancer patients with high KAT6A expression and low KAT6A expression;

[0025] Figure 10 The stacked bar chart for comparing the efficacy of immune checkpoint inhibitor (ICI) treatment in colorectal cancer patients with high KAT6A expression and low KAT6A expression. Detailed implementation manners

[0026] To make the objectives, technical solutions and beneficial effects of the present invention clearer, the technical solutions of the present invention will be described in detail below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. The following gives detailed implementation schemes and specific operation procedures, but the protection scope of the present invention is not limited to the following embodiments.

[0027] Example 1: In vivo targeting of KAT6A combined with anti-PD-1 treatment for CT26 cell syngeneic murine colorectal cancer subcutaneous xenografts.

[0028] The KAT6A inhibitor used in this example is WM-1119 (MCE, HY-102058), which is a commonly used lysine acetyltransferase inhibitor in the field of colorectal cancer research. Other KAT6A inhibitors can also be selected according to experimental or clinical needs.

[0029] The anti-PD-1 monoclonal antibody (immune checkpoint inhibitor) used in this example is InVivoMAb anti-mouse PD-1 (CD279), which is a commonly used anti-PD-1 monoclonal antibody in the field of colorectal cancer research. Other anti-PD-1 monoclonal antibodies can also be selected according to experimental or clinical needs.

[0030] 1. Construction of a murine MSS-type colorectal cancer model.

[0031] (1) Female BALB / c mice at 5-6 weeks of age were used for the construction of the tumor-bearing model;

[0032] (2) After digestion and counting, CT26 cells (murine MSS-type colorectal cancer cells) were inoculated subcutaneously into mice at a density of 1×10 6 cells / 100 μL;

[0033] (3) When the size of the transplanted tumor grew to about 100 mm 3 (about 10 days), group treatment was carried out.

[0034] 2. The constructed murine MSS-type colorectal cancer model mice were divided into 4 groups, with 5 mice in each group.

[0035] Among them, the reagent preparation scheme is as follows: CD279 is prepared with normal saline; WM-1119 is prepared into a stock solution of 10 mg / mL with DMSO, and it is prepared immediately before use and configured according to the ratio of 10% DMSO + 90% (20% SBE-β-CD in saline).

[0036] Among them, the specific grouping is as follows (the grouping and drug administration process of mice with MSS-type colorectal cancer model are as Figure 1 shown):

[0037] ① Control group: Intraperitoneally inject 1M sterile PBS, 100 μL / mouse, once a day;

[0038] ② KAT6A inhibitor group: Intraperitoneally inject WM-1119, 600 μg / mouse, four times a day for a total of eight days;

[0039] ④ anti-PD-1 group: Intraperitoneally inject CD279, 200 μg / mouse, three times a week for a total of four times;

[0040] ⑤ KAT6A inhibitor combined with anti-PD-1 group: Intraperitoneally inject WM-1119, 600 μg / mouse, four times a day for a total of eight days; intraperitoneally inject CD279, 200 μg / mouse, three times a week for a total of four times; the two drugs are treated separately at the same time period.

[0041] 3. Detection of tumor volume:

[0042] Starting from the 10th day after inoculating CT26 cells, every 2 - 3 days, use vernier calipers to measure the length and width of the subcutaneous transplanted tumors in mice, calculate the average tumor diameter, and the statistical results are as Figures 2 - 4 shown.

[0043] Results: As Figures 2 - 4 shown, the single-agent KAT6A inhibitor group inhibited the growth of tumor volume in mice, and there was no significant change in the tumor volume of the single-agent anti-PD-1 group in mice. In addition, the dual-agent combination treatment group significantly inhibited the growth of tumor volume in mice compared with the single-agent treatment group, and at the same time did not cause obvious side effects. The above results suggest that targeting KAT6A effectively enhances the therapeutic efficacy of anti-PD-1.

[0044] Figure 2 There was no difference in the tumor size between the first row and the second row in [], indicating that the single-agent PD-1 was ineffective for subcutaneous transplanted tumors, that is, there was drug resistance. Comparing the KAT6A inhibitor combined with anti-PD-1 group (the fourth row) with the single-agent anti-PD-1 group (the second row), the tumor significantly shrank after anti-PD-1 was combined with the KAT6A inhibitor, that is, it indicated that the problem of drug resistance was solved.

[0045] Example 2: In vivo targeting of KAT6A combined with anti-PD-1 for the treatment of subcutaneous transplanted tumors of MC38 homologous murine colorectal cancer.

[0046] 1. Construction of an in situ model of murine MSI-H colorectal cancer.

[0047] (1) Female C57BL / 6 mice aged 5-6 weeks were used for the construction of the tumor-bearing model;

[0048] (2) MC38 cells (murine MSI-H colorectal cancer cells) after digestion and counting were inoculated subcutaneously into mice at a density of 1×10 6 cells / 100 μL;

[0049] (3) When the size of the transplanted tumor grew to about 100 mm 3 (about 10 days), grouped treatment was carried out.

[0050] 2. The mice with the constructed in situ model of MSI-H colorectal cancer were divided into 4 groups, with 5 mice in each group.

[0051] Among them, the reagent preparation schemes are as follows: CD279 was prepared with normal saline; WM-1119 was prepared into a stock solution of 10 mg / mL with DMSO and was freshly prepared before use and configured according to the ratio of 10% DMSO + 90% (20% SBE-β-CD in saline).

[0052] Among them, the specific grouping is as follows (the grouping and drug administration process of mice with MSI-H colorectal cancer model are as Figure 5 shown):

[0053] ① Control group: Intraperitoneal injection of 1M sterile PBS, 100 μL / mouse, once a day;

[0054] ② KAT6A inhibitor group: Intraperitoneal injection of WM-1119, 600 μg / mouse, four times a day for a total of eight days;

[0055] ④ anti-PD-1 group: Intraperitoneal injection of CD279, 200 μg / mouse, three times a week for a total of four times;

[0056] ⑤ KAT6A inhibitor combined with anti-PD-1 group: Intraperitoneal injection of WM-1119, 600 μg / mouse, four times a day for a total of eight days; Intraperitoneal injection of CD279, 200 μg / mouse, three times a week for a total of four times; The two drugs were treated separately at the same time period.

[0057] 3. Detection of tumor volume:

[0058] Starting from the 10th day after inoculation with MC38 cells, every 2 - 3 days, the length and width of the subcutaneous transplanted tumors in mice were measured using a vernier caliper, and the average tumor diameter was calculated. The statistical results are as Figures 6 - 8 shown.

[0059] Results: As Figures 6 - 8 shown, the single-agent KAT6A inhibitor group inhibited the growth of tumor volume in mice, and the single-agent anti-PD-1 group had a better effect than the single-agent KAT6A inhibitor group. In addition, the dual-agent combination treatment group more significantly inhibited the growth of tumor volume in mice compared with the single-agent treatment group, and no obvious side effects were caused. The above results suggest that targeting KAT6A effectively enhances the therapeutic efficacy of anti-PD-1.

[0060] Example 3: Clinical cohort and analysis

[0061] A total of 103 clinical cohorts of colorectal cancer (CRC) patients receiving ICI treatment were analyzed in the present invention. The efficacy evaluation was based on the Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1: Complete remission (CR) means that all target lesions completely disappear and the short-axis diameter of all pathological lymph nodes is measured as < 10 mm; Partial remission (PR) means that the sum of the diameters of the target lesions is reduced by ≥ 30% compared with the baseline and no new lesions appear; Progressive disease (PD) means that the sum of the diameters of the target lesions is increased by ≥ 20% (and the absolute increase is ≥ 5 mm) compared with the baseline or one or more new lesions appear; Stable disease (SD) means that the criteria of neither PR nor PD are met. The objective response rate (ORR) refers to the proportion of patients achieving CR or PR, and the disease control rate (DCR) refers to the proportion of patients achieving CR, PR, or SD.

[0062] The immunohistochemical staining technique was used to evaluate the KAT6A expression level, and the analysis was carried out according to the IRS scoring system (combining staining intensity and the proportion of positive cells): The staining intensity was scored from 0 to 3 levels (0 points: no staining; 1 point: weak staining; 2 points: medium-intensity staining; 3 points: strong staining), and the proportion of positive cells was scored from 0 to 4 levels (0 points: 0% positive cells; 1 point: 1% - 10%; 2 points: 11% - 50%; 3 points: 51% - 80%; 4 points: 81% - 100%). The final IRS score was obtained by multiplying the scores of the two (total score range 0 - 12 points), and the patients were divided into the KAT6A high-expression group (score ≥ 6 points) and the low-expression group (score < 6 points) with the median as the boundary.

[0063] The waterfall plot comparing the therapeutic effects of colorectal cancer patients with high KAT6A expression and low KAT6A expression receiving immune checkpoint inhibitor (ICI) treatment is as Figure 9 shown, and the stacked bar chart comparing the therapeutic effects of colorectal cancer patients with high KAT6A expression and low KAT6A expression receiving immune checkpoint inhibitor (ICI) treatment is as Figure 10As shown. In this cohort, 49.1% (27 / 55) of the 55 patients with low KAT6A expression achieved a complete response (PR), and 41.8% (23 / 55) had stable disease (SD). The objective response rate (ORR) of the low KAT6A expression group was 49.1% (27 / 55), and the disease control rate (DCR) was 90.9% (50 / 55). Both of these indicators were significantly better than those of the high KAT6A expression group (ORR: 10.4%, 5 / 48; DCR: 52.1%, 25 / 48). The above data indicate that KAT6A is a highly promising therapeutic target, and inhibiting this target can significantly improve the efficacy of immune checkpoint inhibitors in the treatment of colorectal cancer, providing a new biomarker for the immunotherapy of colorectal cancer.

Claims

1. Use of a KAT6A inhibitor in combination with a PD-1 monoclonal antibody in the preparation of a medicament for treating colorectal cancer.

2. The application according to claim 1, wherein The KAT6A inhibitor is WM-1119.

3. The application according to claim 1, characterized in that, The colorectal cancer is MSS-type colorectal cancer and MSI-H type colorectal cancer.

4. The application according to claim 1, wherein The PD-1 monoclonal antibody is CD279.

5. Use of a KAT6A inhibitor in the preparation of a medicament for treating colorectal cancer.

6. The application according to claim 5, wherein The KAT6A inhibitor is WM-1119.

7. The application according to claim 5, wherein The colorectal cancer is MSS-type colorectal cancer and MSI-H type colorectal cancer.

8. A pharmaceutical composition for treating colorectal cancer, characterized in that, The pharmaceutical composition comprises a KAT6A inhibitor and a PD-1 monoclonal antibody.

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