Novel tumor microenvironment immunomodulator and application thereof

Abstract

The invention belongs to the technical field of medicines, and relates to a novel tumor microenvironment immunomodulator and application thereof. The novel tumor microenvironment immunomodulator is composed of FK228 or a derivative thereof and a PD-1 or PD-L1 inhibitor. The derivative of the FK228 is FK228B or FK228C, and the derivative of the FK228 is FK228B or The invention also provides application of the immunomodulator. The immunomodulator is used for preparing a composition for treating solid tumors. The FK228 or the derivative thereof in the novel tumor microenvironment immunomodulator provided by the invention can increase the sensitivity of solid tumors to a PD-(L) 1 inhibitor, and can enhance the treatment of solid tumors with low sensitivity to ICBs such as the PD-(L) 1 inhibitor, so that a new product and a new thought are provided for clinically treating the solid tumors.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology and relates to a novel tumor microenvironment immunomodulator and its application. Background Art

[0002] A favorable tumor immune microenvironment (TIME) helps improve the response rate of human solid tumors to immune checkpoint blockers (ICB). Clinically, many solid tumor patients have low sensitivity to ICB treatment. Therefore, it is necessary to develop new combination treatment strategies. Increasing evidence shows that histone deacetylase (HDAC) plays a positive or negative regulatory role in maintaining immune-unfavorable TIME. Does inhibiting HDAC activity have a positive effect on the regulation of TIME? There is no relevant report yet.

[0003] FK228, also known as romidepsin, is a small molecule drug developed by Celgene Corporation of the United States and first approved by the U.S. Food and Drug Administration (FDA) for the treatment of non-solid tumors such as skin or peripheral T-cell lymphoma in 2009. FK228 is a best HDAC inhibitor (HDACi) that can directly act on cancer cells, activate the expression of epigenetic silencing genes, and inhibit the proliferation of cancer cells.

[0004] However, compared with non-solid tumors such as lymphoma, solid tumors show highly complex TIME, which contains cancer cells, endothelial cells and various immune cells, some of which can promote cancer cell growth and assist cancer cell immune escape. The state of these TIME plays a decisive role in the treatment of solid tumors. It is worth noting that there is no report on FK228 and its new derivatives comprehensively regulating TIME, thereby turning unfavorable TIME into favorable TIME. At the same time, there are no patents and literature reports on the combined administration of FK228 and its new derivatives with ICB to treat solid tumors. Summary of the invention

[0005] The purpose of the present invention is to provide a tumor microenvironment immunomodulator for patients with solid tumors with low sensitivity to ICB treatment. The immunomodulator is composed of FK228 or its derivatives and PD-(L)1 inhibitors. Experimental results show that FK228 and its derivatives can increase the sensitivity of solid tumors to PD-(L)1 inhibitors, and the combination of the two significantly inhibits the growth of solid tumors.

[0006] In order to achieve the above-mentioned purpose, the technical solution adopted by the present invention is: a new tumor microenvironment immunomodulator, composed of FK228 or its derivatives and PD-1 or PD-L1 inhibitors; the derivative of FK228 is FK228B or FK228C.

[0007] Preferably, the PD-1 or PD-L1 inhibitor is a PD-1 or PD-L1 antibody.

[0008] Preferably, the antibody is selected from any one of atezolizumab, envobalizumab, and pembrolizumab.

[0009] Preferably, the dosage of FK228 or its derivatives in the immunomodulator is 2-13 mg / m 2 / Second-rate.

[0010] Preferably, among the immunomodulators, the dosage of the PD-1 inhibitor is 10 to 30 mg / kg / time.

[0011] Preferably, among the immunomodulators, the dosage of the PD-L1 inhibitor is 4 to 7 mg / kg / time.

[0012] The present invention also provides the use of the immunomodulator, which is used for preparing a composition for treating solid tumors.

[0013] Compared with the existing technologies and methods, the beneficial effects of the present invention are: FK228 or its derivatives can increase the sensitivity of solid tumors to PD-(L)1 inhibitors, and can enhance the treatment of solid tumors with low sensitivity to ICBs such as PD-(L)1 inhibitors. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 The single-cell sequencing analysis diagram of FK228 comprehensively regulating the tumor immune microenvironment; A is the experimental model diagram of three transplanted tumor mouse models, B is the single-cell atlas of the tumor tissue microenvironment of the three transplanted tumor mouse models before and after FK228 treatment, C is the difference atlas of T cells in tumor tissue of the three transplanted tumor mouse models before and after FK228 treatment, D is the difference in the abundance of various T cells in tumor tissue of the three transplanted tumor mouse models before and after FK228 treatment, E is the difference atlas of macrophages in tumor tissue of the three transplanted tumor mouse models before and after FK228 treatment, and F is the difference in the abundance of various macrophages in tumor tissue of the three transplanted tumor mouse models before and after FK228 treatment; Figure 2 This is a single-cell sequencing analysis diagram of FK228B comprehensively regulating the tumor immune microenvironment; A is a single-cell map of various cells in tumor tissues treated with FK228B, and B is the difference in the abundance of various immune cells in tumor tissues before and after FK228B treatment; Figure 3The figure shows the therapeutic effect of FK228 combined with PD-L1 inhibitor in treating mouse cold tumor CT26 transplanted tumor; A is the combined treatment mode diagram of FK228 and PD-L1 inhibitor, B is the tumor morphology before and after the combined treatment of FK228 and PD-L1 inhibitor, and C is the tumor proliferation curve before and after the combined treatment of FK228 and PD-L1 inhibitor; Figure 4 This is the body weight chart of mice with cold tumor CT26 xenografts after treatment with FK228 combined with PD-1 inhibitors; Figure 5 Figure 2 is a single-cell sequencing analysis of the tumor immune microenvironment treated with FK228 combined with PD-L1 inhibitors; A is the annotated cell populations before and after the combined treatment with FK228 and PD-L1 inhibitors, B is the difference map of each cell population before and after the combined treatment with FK228 and PD-L1 inhibitors, C is the difference map of T cell subsets before and after the combined treatment with FK228 and PD-L1 inhibitors, D is the difference map of macrophage subsets in the combined treatment with FK228 and PD-L1 inhibitors, E is the difference in abundance of each immune cell before and after the combined treatment with FK228 and PD-L1 inhibitors, F is the difference in abundance of several immune cells before and after the combined treatment with FK228 and PD-L1 inhibitors detected by flow cytometry, G is the difference in abundance of anti-tumor CD8 + Changes in T cell killing toxicity; Figure 6 This is the body weight chart of mice with cold tumor CT26 xenografts treated with FK228B combined with PD-(L)1 inhibitors after treatment; Figure 7 This is the body weight chart of mice with cold tumor CT26 transplanted tumors after treatment with FK228C combined with PD-(L)1 inhibitor. DETAILED DESCRIPTION

[0015] The present invention is further described below in conjunction with specific embodiments and the accompanying drawings. More details are set forth in the following description to facilitate a full understanding of the present invention. However, the present invention is obviously capable of being implemented in a variety of other ways different from the description herein. For those skilled in the art, any substitution, improvement or change made to the embodiments of the present invention is within the protection scope of the present invention, and the protection scope of the present invention should not be limited by the content of this specific embodiment.

[0016] FK228 used in the following examples was purchased from MCE, FK228B and FK228C were produced by our own fermentation and purification (ZL202211122305.0), PD-L1 and PD-1 inhibitors (Envolimab, Pembrolizumab) were purchased from BioXCell, and the remaining cell-related reagents and consumables and animal experiment reagents and consumables were obtained from commercial channels.

[0017] Example 1: Treatment with FK228 and its novel derivative FK228B causes global changes in the immune microenvironment of solid tumors The number is 1×10 5 Mouse colon cancer cells MC38, CT26 and mouse breast cancer cells 4T1 were subcutaneously injected into C57BL / 6 and BALB / C mice. When the tumors grew to 50 mm 3 When the mice were treated with FK228 or FK228B, the mice were injected with 0.5 mg / kg of FK228 or FK228B via the tail vein. The mice were given an equal volume of saline as a control. The injections were repeated every 2 days for a total of 5 times. The tumor tissues were then collected and sent to Beijing Xunyin Biotechnology Co., Ltd. for single-cell transcriptome sequencing. After in-depth analysis of the sequencing data, it was found that after FK228 and FK228B treatment, the immune microenvironment of solid tumors changed significantly, specifically: anti-tumor CD8 + Immune cells such as T cells, pro-inflammatory macrophages, NK cells, and dendritic cells were significantly enriched in tumor tissues, among which CD8 + T cells are significantly activated, while anti-inflammatory macrophages and Treg cells that promote tumors are significantly reduced in tumor tissues. Figure 1 , Figure 2 There is no report on the global characterization of the tumor immune microenvironment after FK228 treatment of solid tumors in previous literature and patents. The data of the present invention discloses for the first time the single-cell map of the tumor immune microenvironment after FK228 treatment of solid tumors. More importantly, the present invention unexpectedly found that the new FK228 derivative FK228B developed by our laboratory can also regulate the immune microenvironment of solid tumors, which will promote the clinical application of the drug.

[0018] Example 2: FK228 combined with PD-(L)1 inhibitor significantly reduced the growth of solid tumors A tumor-bearing mouse model was constructed by referring to the method of Example 1, and then FK228 and PD-L1 or PD-1 inhibitor were injected through the tail vein, with a dose of 0.5 mg / kg and 200 μg / mouse, respectively, once every 2 days, for a total of 5 injections, and the tumor volume was monitored once every 2 days. Then the tumor tissue was taken for photography, and it was found that FK228 could sensitize the response of solid tumors to PD-L1 or PD-1 inhibitors, and the combined treatment group showed excellent therapeutic effects, such as Figure 3 and Figure 4 shown.

[0019] Example 3: Single cell profiling analysis of tumor tissues treated with FK228 combined with PD-L1 inhibitors The tumor in Example 2 was sent to Beijing Xunyin Biotechnology Co., Ltd. for single-cell transcriptome sequencing, which clearly depicted the immune microenvironment map in the tumor tissue before and after combined treatment. Figure 5 As shown, it was found that various immune cells underwent significant changes after combined treatment, especially an increase in anti-tumor immune cells and a decrease in pro-tumor immune cells. This data will further reveal the mechanism of combined treatment and lay the foundation for clinical application.

[0020] Example 4: FK228B combined with PD-(L)1 inhibitor significantly reduced the growth of solid tumors A tumor-bearing mouse model was constructed by referring to the method of Example 1, and then FK228B and PD-L1 or PD-1 inhibitors were injected through the tail vein, with doses of 0.5 mg / kg and 200 μg / mouse, respectively, once every 2 days, for a total of 5 injections. After the treatment, the mice were dissected to obtain the tumors and weighed. It was found that FK228B could also sensitize solid tumors to PD-L1 or PD-1 inhibitors. The combined treatment group showed excellent therapeutic effects, such as Figure 6 shown.

[0021] Example 5: FK228C combined with PD-(L)1 inhibitor significantly reduced the growth of solid tumors A tumor-bearing mouse model was constructed by referring to the method of Example 1, and then FK228C and PD-L1 or PD-1 inhibitors were injected through the tail vein, with doses of 0.5 mg / kg and 200 μg / mouse, respectively, once every 2 days, for a total of 5 injections. After the treatment, the mice were dissected to obtain the tumors and weighed. It was found that FK228C could also sensitize solid tumors to PD-L1 or PD-1 inhibitors. The combined treatment group showed excellent therapeutic effects, such as Figure 7 shown.

Claims

1. A novel tumor microenvironment immunomodulator, characterized in that: It consists of FK228 or its derivatives and PD-1 or PD-L1 inhibitors; the FK228 derivative is FK228B or FK228C.

2. The novel tumor microenvironment immunomodulator according to claim 1, characterized in that: The PD-1 or PD-L1 inhibitor is a PD-1 or PD-L1 antibody.

3. The novel tumor microenvironment immunomodulator according to claim 2, characterized in that: The antibody is selected from any one of atezolizumab, envobalizumab, and pembrolizumab.

4. The novel tumor microenvironment immunomodulator according to any one of claims 1 to 3, characterized in that: In the immunomodulator, the dosage of FK228 or its derivatives is 2-13 mg / m 2 / Second-rate.

5. The novel tumor microenvironment immunomodulator according to any one of claims 1 to 3, characterized in that: Among the immunomodulators, the dosage of the PD-1 inhibitor is 10 to 30 mg / kg / time.

6. The novel tumor microenvironment immunomodulator according to any one of claims 1 to 3, characterized in that: Among the immunomodulators, the dosage of the PD-L1 inhibitor is 4 to 7 mg / kg / time.

7. The use of the novel tumor microenvironment immunomodulator according to any one of claims 1 to 3, characterized in that: The immunomodulator is used for preparing a composition for treating solid tumors.

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