Application of PDE4 inhibitor and PD-L1 inhibitor in treatment of leukemia
By combining PDE4 inhibitors with PD-L1 inhibitors, CD8+ T cells are activated, the proportion of AML cells is reduced, and survival is prolonged, and the problem of poor efficacy of existing anti-PD-1/PD-L1 immunotherapy in AML patients is solved, and effective treatment of AML is achieved.
Patent Information
- Application Number
- CN202311271322.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2023-09-27
- Publication Date
- 2025-03-28
AI Technical Summary
The existing anti-PD-1/PD-L1 immunotherapy has poor efficacy in patients with acute myeloid leukemia (AML), and high-dose chemotherapy has great toxic and side effects on elderly patients and is unbearable.
PDE4 inhibitors are used in combination with PD-L1 inhibitors to enhance antitumor immune response. Specifically, by activating CD8+ T cells, the proportion of AML cells is reduced and the survival of mice is extended using PDE4 inhibitors such as ZL-n-91 or Apst combined with PD-L1 inhibitors such as PD-L1 antibodies.
This combination therapy significantly increased the proportion of CD8+ T cells, reduced the proportion of AML cells, prolonged the survival of AML mice, and had good anti-tumor effects.
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Figure CN119700988A_ABST
Abstract
Description
Technical Field
[0001] The present application belongs to the field of anti-tumor drugs, especially leukemia drugs. Specifically, the present application provides the use of a PDE4 inhibitor combined with a PD-L1 inhibitor to treat leukemia. Background Art
[0002] Acute myeloid leukemia (AML) is a type of bone marrow heterogeneous disease characterized by abnormal proliferation and blocked differentiation of myeloid hematopoietic stem cells. Its etiology and pathogenesis are complex and not yet fully understood. AML infiltrates many important human organs, such as the liver, spleen, and lymph nodes, thereby inhibiting body functions. AML progresses rapidly and is difficult to control. The median age of AML patients at diagnosis is around 65 years old, and the 5-year survival rate of young patients 60 and under is 35-45%; while the survival rate of elderly patients over 60 years old is only 5-15%. In the past 30 years, combined chemotherapy has been the main method for clinical treatment of AML. However, high-dose chemotherapy has very large toxic side effects, which can cause symptoms of bone marrow suppression, pancytopenia, organ damage, and even neurological and cardiac toxicity in AML patients. Elderly AML patients over 60 years old generally cannot withstand high-dose chemotherapy, which can be life-threatening. Therefore, it is of great significance to find new and mild therapies.
[0003] In recent years, anti-PD-1 / PD-L1 immunotherapy has revolutionized cancer treatment and has shown significant clinical efficacy in solid tumors, such as melanoma, non-small cell lung cancer, and pleural mesothelioma. However, compared with solid cancers, anti-PD-1 / PD-L1 immunotherapy has poor efficacy in AML patients, so PD-1 / PD-L1 inhibitors are not approved by the FDA for the treatment of AML. Many researchers have proposed that drugs that enhance antigen presentation and / or increase immune checkpoint expression can be combined with PD-1 / PD-L1 inhibitors to improve the response rate of AML patients to PD-1 / PD-L inhibitors.
[0004] Phosphodiesterase (PDEs) has the function of hydrolyzing intracellular second messengers cAMP or cGMP, thereby affecting the signaling pathways mediated by these second messengers and regulating cell functions. PDEs are divided into 11 subtypes, among which phosphodiesterase 4 (PDE4) specifically hydrolyzes cAMP. It has been reported in the literature that cAMP can increase the expression of CD86 in major histocompatibility complex (MHC) class II to increase the antigen presentation function of dendritic cells. At present, there is no study on the effect of PDE4 inhibitors combined with PD-L1 inhibitors to exert a synergistic tumor-killing effect in the treatment of acute myeloid leukemia. Summary of the invention
[0005] The present invention combines a PDE4 inhibitor with a PD-L1 inhibitor to treat acute myeloid leukemia, providing a new combination therapy for tumor patients.
[0006] On the one hand, the present application provides the use of a PDE4 inhibitor combined with a PD-L1 inhibitor in the preparation of a medicament for treating leukemia.
[0007] Furthermore, the leukemia is acute myeloid leukemia.
[0008] Furthermore, the PDE4 inhibitor is Zl-n-91 or Apremilast.
[0009] Furthermore, the PD-L1 inhibitor is a PD-L1 antibody.
[0010] Furthermore, the PD-L1 antibody is selected from atezolizumab, durvalumab, durvalumab, atezolizumab and BioXcell, BE0101.
[0011] On the other hand, the present application provides a drug for treating leukemia, which comprises a PDE4 inhibitor and a PD-L1 inhibitor.
[0012] Furthermore, the leukemia is acute myeloid leukemia.
[0013] Furthermore, the PDE4 inhibitor is Zl-n-91 or Apremilast.
[0014] Furthermore, the PD-L1 inhibitor is a PD-L1 antibody.
[0015] Furthermore, the PD-L1 antibody is selected from atezolizumab, durvalumab, durvalumab, atezolizumab and BioXcell, BE0101.
[0016] The chemical structures of ZL-n-91 and Apremilast described in the present application have been disclosed. Those skilled in the art can refer to the prior art literature (such as Ruihong Ma, Bin-yan Yang, Chang-you Wu. A selective phosphodiester erase 4 (PDE4) inhibitor Zl-n-91 suppresses IL-17 production by human memory Th17 cells. International Immunopharmacology, 2008, 8 (10): 1408-1417. and Zhao Qian, Sun Yue, Shi Yu, etc. Phosphodiesterase-4 inhibitor Apremilast [J]. Modern Drugs and Clinics, 2014, 29 (04): 428-433.) and conventional techniques in the field of organic chemistry to obtain ZL-n-91 of suitable purity by synthesis, purchase, application for donation, etc.
[0017] The leukemia described in this application includes various specific classifications of leukemia, including but not limited to, acute / chronic leukemia, myeloid / granulocytic / lymphocytic leukemia, and childhood / adult leukemia.
[0018] In addition to the above antibodies, other PD-L1 antibodies that are commercially available or under development can also be routinely selected by those skilled in the art.
[0019] The composition of the present invention is in any clinically acceptable dosage form, including various dosage forms for oral and parenteral administration. For oral administration, it can be tablets, capsules, soft capsules, oral liquids, syrups, granules, dripping pills, orodisintegrating tablets, sustained-release tablets, sustained-release capsules, controlled-release tablets, controlled-release capsules, etc.; for parenteral administration, it can be water injection, lyophilized powder injection, sterile powder injection, infusion, etc.
[0020] The PDE4 inhibitor combined with the PD-L1 inhibitor in the present invention may be in the same or different dosage forms, for example, they may be injection or oral dosage forms respectively. The PDE4 inhibitor combined with the PD-L1 inhibitor in the present invention may be prepared into the same pharmaceutical composition, or they may be provided separately and used in combination under the guidance of the instructions or doctor's advice.
[0021] Those skilled in the art can use various pharmaceutically acceptable carriers or excipients to construct the above-mentioned dosage forms, such as solvents, co-solvents, pH adjusters, osmotic pressure regulators, stabilizers, antioxidants, fillers, binders, lubricants, disintegrants, coating agents, sustained-release agents, controlled-release agents, adsorption carriers, etc.
[0022] The phosphodiesterase 4 inhibitor Apremilast or ZL-n-91 combined with PD-L1 inhibitor can inhibit the proliferation of acute myeloid leukemia cells and have a good anti-tumor effect: compared with the use of phosphodiesterase 4 inhibitors or PD-L1 inhibitors alone, the combination of drugs can more effectively increase CD8 + The proportion of T cells, activated immunity; reduced GFP + -The proportion of AML; effectively prolong the survival time of mice. BRIEF DESCRIPTION OF THE DRAWINGS
[0023] Figure 1 :The PDE4 inhibitor Apremilast combined with PD-L1 inhibitor significantly increased CD8 + The number of T cells. Figure 1 The following parts are as follows: (A) Dosage regimen of PDE4 inhibitor Apremilast combined with PD-L1 inhibitor in the treatment of AML mice; (B) Flow cytometry detection of CD3 in peripheral blood leukocytes of AML mice in each group + T cell population ratio; (C) Flow cytometry detection of CD3 + CD4 in T cells + T and CD8 + T cell population ratio; (D) CD3 + Quantitative analysis of cell populations; (E) CD3 + CD4 + Quantitative analysis of cell populations; (F) CD3 + CD8 + Quantitative analysis of cell populations. Data are presented as mean ± SD, n = 6. *P < 0.05, **P < 0.01 and ****P < 0.0001.
[0024] Figure 2 :The PDE4 inhibitor Apremilast combined with PD-L1 inhibitor significantly reduced the proportion of AML cells in AML mice. (A) Flow cytometry was used to detect GFP in peripheral blood leukocytes of AML mice on days 15 and 21 after modeling. + -Histogram of AML cells; (B) GFP expression in leukocytes of AML mice in each group on day 15 + -Quantitative analysis of AML cells; (C) GFP expression in leukocytes of AML mice in each group on day 21 + - Quantitative analysis of AML cells. Data are presented as mean ± SD, n = 6. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001.
[0025] Figure 3: The PDE4 inhibitor Apremilast combined with PD-L1 inhibitor significantly prolongs the survival of AML mice.
[0026] Figure 4 : Dosing regimen of PDE4 inhibitor ZL-n-91 combined with PD-L1 inhibitor in the treatment of AML mice.
[0027] Figure 5 : The PDE4 inhibitor ZL-n-91 combined with the PD-L1 inhibitor significantly prolongs the survival of AML mice. DETAILED DESCRIPTION
[0028] Example 1 Experiment on the treatment of in situ AML mice with the PDE4 inhibitor Apremilast combined with PD-L1 inhibitor
[0029] (1) After taking out the frozen MLL-AF9-driven AML mouse spleen cells from the liquid nitrogen tank, quickly place them in a 37°C water bath and gently shake them to quickly melt the cell freezing solution;
[0030] (2) Wash twice with PBS, count the cells, and inject into 40 male C57BL / 6 mice. The final injection volume is about 5*10 5 / 200μl / mouse; administration started on the 6th day after injection;
[0031] (3) The mice were divided into four groups (10 mice in each group): solvent control group, phosphodiesterase 4 inhibitor apremilast treatment group (0.5 mg kg -1 , intraperitoneal injection), PD-L1 inhibitor group (purchased from BioXcell, BE0101, 150 μg / mouse / time, intraperitoneal injection), and combined drug group. Figure 1 As shown in Part A.
[0032] (4) On the 19th day after modeling, 6 mice were randomly selected from each group, and about 50ul of blood was collected from the eye sockets. The blood samples were first treated with red blood cell lysis buffer; then anti-mouse CD16 / CD32 (BD, 553141) antibodies were added and incubated for 10 minutes; then anti-mouse CD3-APC-Cy7 (BD, 557596), anti-mouse CD4-PE (BD, 553652) and anti-mouseCD8-PerCP-Cy5.5 (BD, 551162) antibodies were added respectively, and incubated in the dark for 30 minutes. The CD3 in the blood samples was analyzed by FACSDxp AthenaTM (Cytek, USA) instrument. + , CD4 + and CD8 + The proportion of T cells.
[0033] (5) On days 15 and 21 after modeling, 6 mice were randomly selected from each group and 50 μl of blood was collected from the eye sockets. The blood samples were first treated with red blood cell lysis buffer and then analyzed for GFP in the blood samples using the FACSDxp AthenaTM (Cytek, USA) instrument. + -Proportion of AML cells.
[0034] (6) Observe the living conditions of mice every day and record their survival time.
[0035] The results are as follows Figure 1 As shown: In the peripheral blood leukocytes of each group of mice, CD3 + T cells ( Figure 1 D part) and CD4 + T( Figure 1 Compared with the CTRL group, PD-L1 inhibitor alone significantly increased the CD8 + The proportion of T cells ( Figure 1 The F part of the study), while the phosphodiesterase 4 inhibitor apremilast combined with PD-L1 inhibitor treatment group further increased CD8 + The proportion of T cells ( Figure 1 The difference was statistically significant (part F). This indicates that the combination of Apremilast and PD-L1 monoclonal antibody activated adaptive immunity, thereby enhancing anti-tumor activity.
[0036] The results are as follows Figure 2 Shown: On the 15th day after modeling, GFP expression in the Apremilast treatment group or PD-L1 monoclonal antibody treatment group was higher than that in the CTRL group. + -There was no significant difference in the number of AML cells, but in the apremilast and PD-L1 monoclonal antibody combination treatment group, GFP + -A significant reduction in AML cells ( Figure 2 On the 21st day after modeling, although the GFP expression in the Apremilast group was significantly higher than that in the CTRL group, + -AML cells still did not show significant differences, but GFP in the PD-L1 mAb-treated mice + -Significant reduction in AML cells ( Figure 2 A and C parts), and the combined treatment further reduced GFP + -The proportion of AML( Figure 2 A and C of the .
[0037] The results are as follows Figure 3As shown: the median survival time of the solvent control group was 23 days, the median survival time of the phosphodiesterase 4 inhibitor apremilast group was 24 days, the median survival time of the PD-L1 inhibitor group was 24 days, and the median survival time of the combination group was 27 days. Compared with the solvent control group, the combined treatment group significantly prolonged the survival time of AML mice, which was statistically significant.
[0038] The above research results show that the phosphodiesterase 4 inhibitor Apremilast combined with the PD-L1 inhibitor used in the present invention can inhibit the proliferation of acute myeloid leukemia cells and has a good anti-tumor effect.
[0039] Example 2 Experiment on the treatment of in situ AML mice with PDE4 inhibitor ZL-n-91 combined with PD-L1 inhibitor
[0040] (1) After taking out the frozen MLL-AF9-driven AML mouse spleen cells from the liquid nitrogen tank, quickly place them in a 37°C water bath and gently shake them to quickly melt the cell freezing solution;
[0041] (2) Wash twice with PBS, count the cells, and inject into 40 male C57BL / 6 mice. The final injection volume is about 5*10 5 / 200μl / mouse; administration started on the 3rd day after injection;
[0042] (3) The mice were divided into four groups (10 mice in each group): solvent control group, phosphodiesterase 4 inhibitor ZL-n-91 administration group (0.5 mg kg -1 , intraperitoneal injection), PD-L1 inhibitor group (purchased from BioXcell, BE0101, 100 μg / mouse / time, intraperitoneal injection), and combined drug group. Figure 4 shown.
[0043] (4) Observe the living conditions of the mice every day and record their survival period.
[0044] The results are as follows Figure 5 As shown: the median survival time of the solvent control group was 23 days, the median survival time of the phosphodiesterase 4 inhibitor Zl-n-91 administration group was 23 days, the median survival time of the PD-L1 inhibitor administration group was 24.5 days, and the median survival time of the combined drug group was 29 days. Compared with the solvent control group, the combined treatment group significantly prolonged the survival time of AML mice, which was statistically significant.
[0045] The above research results show that the phosphodiesterase 4 inhibitor ZL-n-91 combined with the PD-L1 inhibitor used in the present invention can inhibit the proliferation of acute myeloid leukemia cells and has a good anti-tumor effect.
Claims
1. Use of a PDE4 inhibitor combined with a PD-L1 inhibitor in the preparation of a drug for treating leukemia.
2. The use according to claim 1, wherein the leukemia is acute myeloid leukemia.
3. The method according to claim 1 or 2, wherein the PDE4 inhibitor is Zl-n-91 or Apremilast.
4. The use according to any one of claims 1 to 3, wherein the PD-L1 inhibitor is a PD-L1 antibody.
5. The use according to claim 4, wherein the PD-L1 antibody is selected from atezolizumab, durvalumab, durvalumab, atezolizumab and BioXcell, BE0101.
6. A drug for treating leukemia, comprising a PDE4 inhibitor and a PD-L1 inhibitor.
7. The medicament according to claim 6, wherein the leukemia is acute myeloid leukemia.
8. The drug according to claim 6 or 7, wherein the PDE4 inhibitor is Zl-n-91 or Apremilast.
9. The drug according to any one of claims 6 to 8, wherein the PD-L1 inhibitor is a PD-L1 antibody.
10. The medicament according to claim 9, wherein the PD-L1 antibody is selected from atezolizumab, durvalumab, durvalumab, atezolizumab and BioXcell, BE0101.
11. The use according to any one of claims 1 to 5 or the medicament according to any one of claims 6 to 10, wherein the medicament is in oral and / or injectable dosage form.
12. The use or medicine according to claim 11, wherein the medicine further comprises a pharmaceutically acceptable excipient.
13. The use or medicine according to claim 12, wherein the pharmaceutically acceptable excipient is selected from one or more of solvents, cosolvents, pH regulators, osmotic pressure regulators, stabilizers, antioxidants, fillers, binders, lubricants, disintegrants, coating agents, sustained-release agents, controlled-release agents, and adsorption carriers.