Pharmaceutical composition and use thereof

Abstract

Provided are a pharmaceutical composition and use thereof. Specifically provided is a pharmaceutical composition X, which comprises a compound represented by formula A or a pharmaceutically acceptable salt thereof and a PD-1 monoclonal antibody. The pharmaceutical composition exhibits good safety and significant synergistic advantages, and can effectively treat solid tumors.

Description

A pharmaceutical composition and its application

[0001] This application claims priority to Chinese patent application 2025104633393, filed on April 14, 2025. The entire contents of the aforementioned Chinese patent application are incorporated herein by reference. Technical Field

[0002] This invention relates to a drug combination and its application. Background Technology

[0003] PTEN (Phosphatase and Tensin Homolog) is an important tumor suppressor gene that encodes a tumor suppressor protein with both lipid phosphatase and protein phosphatase activities. It inhibits the PI3K / AKT signaling pathway by dephosphorylating phosphoinositide 3-kinase (PI3K), a pathway that plays a crucial role in cell growth, proliferation, survival, and metabolism. Mutations or deletions in the PTEN gene lead to persistent activation of the PI3K / AKT signaling pathway, thereby promoting tumor cell growth, proliferation, and survival. PTEN mutations are common in various solid tumors, particularly breast cancer, gastric cancer, endometrial cancer, cervical cancer, ovarian cancer, and lung cancer. These mutations result in persistent activation of the PI3K signaling pathway, promoting cell proliferation, survival, and migration, thus driving tumor development and progression.

[0004] PI3K constitutes a family of lipid kinases involved in the regulation of signal transduction pathways controlling a range of cellular processes. Based on their substrate specificity, PI3K is divided into three distinct subfamilies: class I, class II, and class III. Class IA PI3K possesses a p110α, p110β, or p110δ catalytic subunit that is complexed with one of the three regulatory subunits p85α, p85β, or p55δ. Class IA PI3K is activated via receptor tyrosine kinases, antigen receptors, G-protein coupled receptors (GPCRs), and cytokine receptors. Class IA PI3K primarily produces phosphatidylinositol-3,4,5-triphosphate (PI(3,4,5)P3), which is a second messenger activating the downstream target AKT. The consequences of AKT bioactivation include tumor cell progression, tumor cell proliferation, tumor cell survival, and growth. There is substantial evidence that the PI3K / AKT pathway is aberrantly regulated in many human cancers, and PI3K pathway activation is also closely associated with alterations in the tumor microenvironment and immune escape. Furthermore, PI3K activity has been involved in endocrinology, cardiovascular disease, immune dysregulation, and inflammation. PI3K-p110δ has been identified as playing a crucial role in the recruitment and activation of immune cells and inflammatory cells. PI3K-p110δ is also upregulated in many human tumors and plays a key role in tumor cell proliferation and survival.

[0005] PD-1 (Programmed Cell Death Protein 1) is an immune checkpoint protein mainly expressed by T cells. PD-L1 (PD-1 Ligand 1) is the main ligand of PD-1, which is widely expressed in tumor cells and immune cells in the tumor microenvironment. PD-1 inhibits the activation and proliferation of T cells by binding to PD-L1 or PD-L2, thereby inhibiting the anti-tumor immune response. PD-1 mAb (e.g., pembrolizumab, nivolumab, etc.) can block this inhibition, helping to restore T cell immune attack on tumor cells. In PTEN-mutated tumors, PD-L1 expression is usually high.

[0006] Currently, there is no approved therapy and drug for solid tumors with PTEN deletion, and it is urgent to develop a combination drug of PI3K inhibitor and PD-1 mAb, so as to bring a breakthrough for the treatment of PTEN-mutated solid tumors and meet the urgent treatment needs of these patient groups. SUMMARY

[0007] The technical problem to be solved by the present application is that there is no approved therapy and drug for solid tumors with PTEN deletion, and therefore a pharmaceutical composition and its application are provided. The present application combines the compound represented by formula A or its pharmaceutically acceptable salt with PD-1 mAb, which has good safety, and the combination shows synergistic effect, which can provide preclinical data support and scientific basis for the development of clinical trials.

[0008] The present application provides a pharmaceutical composition X comprising a compound represented by formula A or its pharmaceutically acceptable salt and PD-1 mAb;

[0009] In one implementation scheme, the PD-1 monoclonal antibody is selected from pembrolizumab, nivolumab, tislelizumab, toripalimab, penpulimab, zimberelimab, serplulimab, cadonilimab, and pucotelimab. One or more of sintilimab, camrelizumab, and enlonstobart, such as one or more of pembrolizumab, nivolumab, and tislelizumab, or pembrolizumab, nivolumab, or tislelizumab.

[0010] In one embodiment, the pharmaceutical composition X is a compound as shown in Formula A and pembrolizumab.

[0011] In one embodiment, the pharmaceutical composition X is a compound as shown in Formula A and tislelizumab.

[0012] In one embodiment, the drug combination X is a compound as shown in Formula A and nivolumab.

[0013] In one embodiment, the drug combination X is a compound as shown in Formula A and slulimab.

[0014] In the drug combination X, the PD-1 monoclonal antibody and compound A or its pharmaceutically acceptable salt may be administered simultaneously or separately, for example, separately. Separate administration may also be sequential.

[0015] In one embodiment, the pharmaceutical composition X contains the compound of formula A or a pharmaceutically acceptable salt thereof in an MC solution.

[0016] In one embodiment, the MC solution is an MC solution with a mass concentration of 0.5 g / 100 mL. In another embodiment, in the pharmaceutical composition X, the mass concentration of the compound of formula A or a pharmaceutically acceptable salt thereof in the MC solution is 0.1-50 mg / mL, for example, 5 mg / mL.

[0017] In one embodiment, the PD-1 monoclonal antibody is present in a physiological saline solution in the pharmaceutical composition X.

[0018] In one embodiment, the concentration of the PD-1 monoclonal antibody in the physiological saline solution of the pharmaceutical composition X is 0.1-20 mg / mL, for example, 1 mg / mL.

[0019] In one embodiment, the compound of formula A or a pharmaceutically acceptable salt thereof may be administered 1-2 times per day, for example, once per day, in the pharmaceutical composition X.

[0020] In one embodiment, the PD-1 monoclonal antibody in the pharmaceutical composition X may be administered every 7-31 days, for example, every 14 days.

[0021] In one embodiment, the PD-1 monoclonal antibody in the pharmaceutical composition X may be administered twice per week.

[0022] In this invention, the pharmaceutical composition X may further comprise pharmaceutical excipients.

[0023] In this invention, the active ingredients in the pharmaceutical composition X are the compound represented by formula A or its pharmaceutically acceptable salt and the PD-1 monoclonal antibody.

[0024] In this invention, the mass ratio of the compound of formula A or its pharmaceutically acceptable salt to the PD-1 monoclonal antibody in the pharmaceutical composition X can be (2-8):1, for example 5:1.

[0025] In this invention, the content of the compound represented by formula A or its pharmaceutically acceptable salt in the pharmaceutical composition X may be 50-400 mg, for example 50 mg.

[0026] In this invention, the content of the PD-1 monoclonal antibody in the pharmaceutical composition X is 10-500 mg, for example 10 mg.

[0027] In this invention, the dosage of the pharmaceutical composition X is conventional in the art, and those skilled in the art can adjust it according to the condition or the target of administration, for example, according to the weight of the target of administration. The dosage of the compound represented by Formula A or its pharmaceutically acceptable salt may be 4.16-33.3 mg / kg, for example 4.16 mg / kg; the dosage of the PD-1 monoclonal antibody may be 0.83-41.6 mg / kg, for example 0.83 mg / kg.

[0028] In this invention, the pharmaceutical composition X can be administered to patients via various routes, including but not limited to gastrointestinal administration (e.g., oral) or injection (e.g., intramuscular, subcutaneous, or intravenous), which can be adjusted as needed by those skilled in the art, with oral administration being preferred.

[0029] In this invention, the pharmaceutical composition X can be formulated into various suitable dosage forms depending on the route of administration, such as tablets (e.g., sugar-coated tablets, film-coated tablets, enteric-coated tablets, or compressed tablets), capsules (e.g., hard capsules or soft capsules), granules, powders, pills, powders, suspensions, oral liquids, injections, sprays, or drops, with tablets being the preferred dosage form.

[0030] The present invention also provides a pharmaceutical composition Y comprising a first pharmaceutical composition and a second pharmaceutical composition;

[0031] The first pharmaceutical composition comprises the compound shown in Formula A above or a pharmaceutically acceptable salt thereof and pharmaceutical excipients;

[0032] The second pharmaceutical composition comprises the aforementioned PD-1 monoclonal antibody and pharmaceutical excipients.

[0033] In this invention, the first pharmaceutical composition and the second pharmaceutical composition may be administered simultaneously or separately, for example, separately. Separate administration may also mean sequential administration.

[0034] In one embodiment, the first pharmaceutical composition further includes an MC solution.

[0035] In one embodiment, the first pharmaceutical composition comprises the compound represented by Formula A or a pharmaceutically acceptable salt thereof and an MC solution.

[0036] In one embodiment, the second pharmaceutical composition further includes physiological saline.

[0037] In one embodiment, the second pharmaceutical composition comprises a PD-1 monoclonal antibody and saline solution.

[0038] In one embodiment, the first pharmaceutical composition may be administered 1-2 times per day, for example, once per day.

[0039] In one embodiment, the second pharmaceutical composition may be administered every 7-31 days, for example, every 14 days.

[0040] In one embodiment, the second pharmaceutical composition may be administered twice a week.

[0041] In this invention, the mass ratio of the compound of formula A or its pharmaceutically acceptable salt in the first pharmaceutical composition to the PD-1 monoclonal antibody in the second pharmaceutical composition is the same as the mass ratio described in pharmaceutical composition X.

[0042] In this invention, the content of the compound of formula A or its pharmaceutically acceptable salt in the first pharmaceutical composition and the content of the PD-1 monoclonal antibody in the second pharmaceutical composition are the same as those in pharmaceutical composition X.

[0043] In this invention, the first pharmaceutical composition can be administered to patients via various routes, including but not limited to gastrointestinal administration (e.g., oral) or injection (e.g., intramuscular, subcutaneous, or intravenous), which can be adjusted as needed by those skilled in the art, with oral administration being preferred.

[0044] In this invention, the second pharmaceutical composition can be administered to patients via various routes, including but not limited to gastrointestinal administration (e.g., oral) or injection (e.g., intramuscular, subcutaneous, or intravenous), which can be adjusted as needed by those skilled in the art, with injection being preferred.

[0045] In this invention, the first pharmaceutical composition can be formulated into a suitable dosage form according to the method of administration, such as tablets (e.g., sugar-coated tablets, film-coated tablets, enteric-coated tablets, or compressed tablets), capsules (e.g., hard capsules or soft capsules), granules, powders, pills, powders, suspensions, oral liquids, injections, sprays, or drops, etc.; tablet dosage form is preferred.

[0046] In this invention, the second pharmaceutical composition can be formulated into a suitable dosage form according to the method of administration, such as tablets (e.g., sugar-coated tablets, film-coated tablets, enteric-coated tablets, or compressed tablets), capsules (e.g., hard capsules or soft capsules), granules, powders, pills, powders, suspensions, oral liquids, injections, sprays, or drops, etc.; preferably oral dosage forms or injection dosage forms, and more preferably injection dosage forms.

[0047] In this invention, the dosage of the pharmaceutical composition Y is conventional in the art, and those skilled in the art can adjust it according to the condition or the recipient, for example, according to the weight of the recipient. The dosage of the first pharmaceutical composition can be 25-400 mg / person; the dosage of the second pharmaceutical composition is 1-40 mg / kg, which can be 10-40 mg / kg.

[0048] The present invention also provides the use of substance M in the preparation of a medicament for the prevention and / or treatment of tumors; wherein, substance M is the above-mentioned pharmaceutical composition X or the above-mentioned pharmaceutical composition Y.

[0049] The tumor is a solid tumor. The solid tumor is selected from one or more of the following: breast cancer, gastric cancer, endometrial cancer, cervical cancer, ovarian cancer, and lung cancer, such as breast cancer.

[0050] The solid tumors mentioned are locally advanced or metastatic advanced solid tumors with PTEN deficiency or low expression.

[0051] The locally advanced or metastatic solid tumors with PTEN deficiency or low expression are selected from one or more of breast cancer, gastric cancer, endometrial cancer, cervical cancer, ovarian cancer, and lung cancer, such as breast cancer.

[0052] Terminology Explanation:

[0053] The term "pharmaceutically acceptable salt" refers to a salt formed by the compound and a pharmaceutically acceptable inorganic or organic acid, wherein the inorganic acid includes, but is not limited to: hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid, and sulfuric acid; and the organic acid includes, but is not limited to: formic acid, acetic acid, propionic acid, succinic acid, 1,5-naphthalenedisulfonic acid, linalool, oxalic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, valeric acid, diethylacetic acid, malonic acid, succinic acid, fumaric acid, pimelic acid, adipic acid, maleic acid, malic acid, sulfamic acid, phenylpropionic acid, gluconic acid, ascorbic acid, nicotinic acid, isonicotinic acid, methanesulfonic acid, p-toluenesulfonic acid, citric acid, and amino acids;

[0054] The term "pharmaceuticalally acceptable" refers to a substance that is suitable for human use without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), i.e., a substance with a reasonable benefit / risk ratio.

[0055] The term "therapeutic effective amount" refers to the amount of compound administered to a patient that is sufficient to effectively treat the disease. Therapeutic effective amount will vary depending on the compound, the type of disease, the severity of the disease, the patient's age, etc., but may be adjusted as appropriate by those skilled in the art.

[0056] The term "pharmaceutical excipients" refers to the excipients and additives used in the production of pharmaceuticals and the dispensing of prescriptions. It includes all substances contained in pharmaceutical preparations, excluding the active ingredient. See the Pharmacopoeia of the People's Republic of China (2020 edition) or the Handbook of Pharmaceutical Excipients (Raymond C. Rowe, 2009) for details.

[0057] The term “treatment” refers to a therapeutic approach. When a specific condition is involved, treatment means: (1) alleviating one or more biological manifestations of the disease or condition; (2) interfering with (a) one or more points in a biological cascade that causes or precipitates the condition or (b) one or more biological manifestations of the condition; (3) improving one or more symptoms, effects or side effects associated with the condition, or one or more symptoms, effects or side effects associated with the condition or its treatment; or (4) slowing the development of the condition or one or more biological manifestations of the condition.

[0058] The term “prevention” refers to a reduced risk of acquiring or developing a disease or disorder (i.e., resulting in at least one clinical symptom of the disease in a subject who may have been exposed to a disease-causing agent or a pre-existing predisposition to the disease).

[0059] The term "active ingredient" refers to the active ingredient in the pharmaceutical composition of the present invention, namely the compound represented by formula A or a pharmaceutically acceptable salt thereof, and the PD-1 monoclonal antibody.

[0060] Without violating common sense in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0061] The reagents and raw materials used in this invention are all commercially available.

[0062] The positive and progressive effects of this invention are as follows: the tumor growth inhibition rates of the combination drug groups of compound A + nivolumab, compound A + pembrolizumab, compound A + tislelizumab, and compound A + slulimab were 107.30%, 115.63%, 98.67%, and 110.63%, respectively. Compared with the monotherapy groups, the drug composition of this invention can effectively treat solid tumors and has a good synergistic effect. Moreover, in the experimental stage, no acute adverse reactions were observed in the subjects after combination therapy. Attached Figure Description

[0063] Figure 1 shows the changes in body weight of the experimental animals after drug administration.

[0064] Figure 2 shows the GVHD scores of the experimental animals after drug administration.

[0065] Figure 3 shows the changes in tumor volume in experimental animals after drug administration.

[0066] Figure 4 shows the tumor weight of the experimental animals at the end of the experiment.

[0067] Figure 5 shows the results of HCD45 FACS detection in peripheral blood of experimental animals at the end of the experiment. Detailed Implementation

[0068] The present invention will be described in detail below with reference to embodiments, but this does not imply any adverse limitation on the invention. The present invention has been described in detail, and specific embodiments thereof have been disclosed. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present invention without departing from the concept and scope of the invention. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product specification.

[0069] 1. Test drug

[0070] Test drug 1: Compound A.

[0071] Investigational drugs 2: Nivolumab, Pembrolizumab, Tislelizumab, and Slulizumab injection.

[0072] 2. Preparation of the test drug

[0073] 2.1 Solvent selection: Nivolumab, Pembrolizumab, Tislelizumab, and Slulizumab injections: normal saline;

[0074] Compound A: 0.5% MC solution.

[0075] 2.2 Preparation method:

[0076] Table 1 Preparation and storage of test substances

[0077] 3. Experimental equipment and reagents

[0078] Table 2 Experimental Equipment

[0079] Table 3 Experimental Reagents

[0080] Table 4. Antibody Information Detected by Flow Cytometry

[0081] Table 5 PBMC Information

[0082] 4. Experimental Methods

[0083] 4.1 Cell Culture

[0084] 4.1.1 Culture of MDA-MB-468 cells

[0085] MDA-MB-468 cells (human breast cancer cell line, provided by Sixin Cell Bank) were cultured in vitro in Leibovitz's medium supplemented with 10% FBS and 1% PS (37°C, 0% CO2), and passaged 2-3 times per week. Cells in the exponential growth phase were collected and resuspended in a suspension of PBS and Matrigel (PBS to Matrigel volume ratio 1:1) to obtain a cell concentration of 50 × 10⁻⁶ cells / year. 6 A cell suspension of 1 / mL was used to inoculate mice.

[0086] 4.1.2 Processing of PBMC

[0087] After PBMC resuscitation, the cells were resuspended in PBS to achieve a cell concentration of 50 × 10⁻⁶ cells / mL. 6 / mL of cell suspension was used for HuPBMC model reconstruction.

[0088] 4.2 Cell Seeding

[0089] 4.2.1 MDA-MB-468 cell seeding

[0090] NOG mice were used for tumor-bearing experiments. 0.2 mL of MDA-MB-468 tumor cells (10 × 10⁻⁶) were subcutaneously injected into the right back of each mouse. 6 ).

[0091] 4.2.2 HuPBMC cell seeding

[0092] Human PMBCs were used to reconstitute the immune system in NOG mice with severe immunodeficiency. On day 14 after inoculation with MDA-MB-468 cells, the tumor volume of the MDA-MB-468 tumor-bearing mice was measured. 110 mice were selected according to the size of the tumor for PBMC inoculation.

[0093] Each mouse was injected via tail vein with 0.1 mL of PBMC suspension (5 × 10⁻⁶). 6 ), to carry out immune reconstitution.

[0094] 4.3 Grouping and Dosing

[0095] When the tumor volume of PBMC-reconstructed MDA-MB-468 tumor-bearing mice reached 137 mm... 3 Around 110 mice were used to measure tumor volume. Based on tumor volume, 80 mice were randomly divided into 10 groups (n=8) for drug efficacy experiments and drug administration was started. The day of drug administration was defined as Day 0. Specific group information and drug administration information are shown in Table 6.

[0096] Table 6 Experimental Groups Note: n aNumber of animals per group; Dosage volume b Adjust the dosage according to body weight at 10 mL / kg. Discontinue the drug when the mouse's body weight drops by more than 15%, and resume the drug administration when the body weight drops back to within 10%.

[0097] 4.4 Tumor Measurement

[0098] The tumor size was measured using vernier calipers twice a week, with tumor volume expressed in mm. 3 The formula for calculating tumor volume is: V = 0.5 × a × b 2 Where a and b are the long and short diameters of the tumor, respectively. The tumor size is then used to calculate the relative tumor growth rate (T / C%). The T / C% value is calculated using the formula: T / C% = (T... i -T0) / (V i -V0)×100%, T i V is the average tumor volume of the treatment group on a certain day, T0 is the average tumor volume of the treatment group on the first day of treatment (before drug administration), and V is the average tumor volume of the treatment group on the first day of treatment. i To measure T i The average tumor volume of the control group on that day, V0 is the average tumor volume of the control group on the first day of treatment.

[0099] Calculate the TGI (%) for each group using the formula: Tumor Growth Inhibition Rate (TGI) (%) = 100% - T / C (%).

[0100] 4.5G VHD rating

[0101] After huPBMC cell inoculation, the basic survival status of mice was observed daily, and the body weight of mice was measured twice a week. Clinical scores (body weight, mouse posture, activity, fur, skin, etc.) were calculated based on clinical observations. The GVHD scoring criteria are shown in Table 7.

[0102] Table 7 Clinical Scoring Criteria for GVHD

[0103] 4.6 Sample Collection

[0104] As per experimental requirements, this experiment concluded on day 16 after drug administration. At the experimental endpoint, whole blood was collected, and human CD45 levels were detected by flow cytometry (FACS). + Cell (hCD45 cell) ratio. The tumor was removed, weighed, and photographed.

[0105] 5. Statistical Analysis

[0106] Experimental data are presented as mean and standard error (SEM), including tumor volume and mouse weight at each time point. All data were statistically analyzed using GraphPad Prism8. One-way ANOVA was used for intergroup comparisons, with p < 0.05 indicating statistical significance.

[0107] The combined effect is evaluated using the formula: Q = E A+B / (E A +E B -E A *E B ), where E A E represents the tumor growth inhibition rate of drug A alone. B E represents the tumor growth inhibition rate of drug B alone. A+B The tumor growth inhibition rate of the combined use of A and B drugs is represented by Q<0.85, which indicates antagonism; Q≤1.15 indicates addition; and Q>1.15 indicates synergy.

[0108] 6. Test Results

[0109] 6.1 Effects on body weight and GVHD score

[0110] The effects of drug treatment on animal body weight during administration are shown in Table 8 and Figure 1. The GVHD scores of mice in each group after PBMC inoculation are shown in Figure 2.

[0111] Compared with the solvent control group, the weight gain of animals in all treatment groups was slowed. Peripheral blood flow cytometry results are shown in Figure 5. The proportion of hCD45+ increased in both the compound A monotherapy group and the combination therapy group (vs. solvent control, P < 0.05, P < 0.01). Therefore, the slowed drug-related weight gain was caused by GvHD induced by PBMC humanization, and this significant change suggests that compound A has the effect of activating immunity and promoting immune system reconstruction.

[0112] Table 8. Effects of the test substances on animal body weight in the PBMC humanized MDA-MB-468 xenograft model.

[0113] 6.2 Tumor growth inhibition results

[0114] In this experiment, the tumor growth curves and tumor weight results of each group of animals are shown in Table 9, Figure 3 and Figure 4.

[0115] Fifteen days after administration, the tumor volumes in the solvent control group, nivolumab monotherapy group, pembrolizumab monotherapy group, tislelizumab monotherapy group, slulizumab monotherapy group, compound A monotherapy group, compound A + nivolumab combination therapy group, compound A + pembrolizumab combination therapy group, compound A + tislelizumab combination therapy group, and compound A + slulizumab combination therapy group were 340.04 ± 13.15 mm. 3 253.49±12.92mm 3 237.24±9.85mm 3 268.49±17.51mm 3 245.11±11.37mm 3 ,199.67±14.45mm3, 122.54±9.33mm 3 105.66±7.53mm 3 140.04±9.43mm 3 and 115.66±5.41mm 3 Compared with the control group, the tumor volume of each group was significantly reduced (P<0.001), and the tumor growth inhibition rate (%TGI) was 42.73%, 50.75%, 35.33%, 46.80%, 69.21%, 107.30%, 115.63%, 98.67%, and 110.63%, respectively. Compared with compound A and monoclonal antibody monotherapy groups, the combination therapy groups significantly improved antitumor activity (P<0.01). The Q values ​​of each combination therapy group were 1.30, 1.36, 1.23, and 1.32, respectively, all >1.15, suggesting that the combination therapy can produce a synergistic effect compared with the monotherapy groups.

[0116] At the end of the experiment, the tumor weights of each group were 228.4±48.1 mg, 180.0±45.5 mg, 166.3±15.59 mg, 181.3±16.43 mg, 181.3±52.6 mg, 161.8±14.0 mg, 112.0±15.0 mg, 106.6±6.30 mg, 112.3±7.09 mg, and 111.6±20.1 mg, respectively, and the trend of change was similar to that of tumor volume.

[0117] Table 9. Antitumor activity of the test substances in the PBMC humanized MDA-MB-468 xenograft model *P<0.05, **P<0.01, ***P<0.001, vs. group 1; #P<0.05; ##P<0.01, vs. group 2; $$P<0.01, vs. group 3; &&P<0.01, vs. group 4; ^^P<0.01, vs. group 10; @@P<0.001, vs. group 5.

[0118] 6.3 Peripheral blood flow cytometry results

[0119] The experimental endpoint was determined by measuring the proportion of hCD45 in the peripheral blood of mice to determine the PBMC reconstruction rate. The results are shown in Figure 5.

[0120] The results showed that 20 days after PBMC inoculation, the hCD45 percentage in the solvent control group was approximately 10%, indicating successful establishment of the humanized model. Compared to the solvent control group, the hCD45 percentage increased in both the compound A monotherapy group and the compound A + monoclonal antibody combination group, suggesting that compound A promotes the proliferation of human lymphocytes in mouse blood.

[0121] Conclusion: Compound A, nivolumab, pembrolizumab, tislelizumab, and slulimumab, as well as in combination, all exhibited significant antitumor activity in the hPBMC humanized MDA-MB-468 xenograft mouse model. Furthermore, the combination with compound A significantly enhanced antitumor activity compared to the single-agent groups or the compound A monotherapy groups. The enhanced antitumor activity from combination with a PD-1 inhibitor may be related to compound A's role in promoting humanization and increasing the proportion of human lymphocytes in the blood.

Claims

1. A pharmaceutical composition X, characterized in that, It comprises a compound as shown in Formula A or a pharmaceutically acceptable salt thereof and a PD-1 monoclonal antibody; 2. The pharmaceutical composition X according to claim 1, characterized in that, It meets one or more of the following conditions: (1) The active ingredient of the pharmaceutical composition X comprises the compound of formula A or a pharmaceutically acceptable salt thereof and a PD-1 monoclonal antibody; (2) In the pharmaceutical composition X, the PD-1 monoclonal antibody and compound A or a pharmaceutically acceptable salt thereof are administered simultaneously or separately, for example, separately; (3) In the pharmaceutical composition X, the PD-1 monoclonal antibody is selected from one or more of pembrolizumab, nivolumab, tislelizumab, toripalimab, penaprilimab, cepalimab, slulizumab, cantuniginemab, putelizumab, sintilimab, camrelizumab, and endograft; for example, selected from one or more of pembrolizumab, nivolumab, and tislelizumab, or for example, pembrolizumab, nivolumab, or tislelizumab; (4) In the pharmaceutical composition X, the content of the compound represented by formula A or its pharmaceutically acceptable salt is 50-400 mg, for example 50 mg; (5) In the pharmaceutical composition X, the content of the PD-1 monoclonal antibody is 10-500 mg, for example 10 mg; (6) The pharmaceutical composition X further comprises pharmaceutical excipients; (7) The pharmaceutical composition X is administered according to the weight of the recipient, and the dosage of the compound of formula A or its pharmaceutically acceptable salt is 4.16-41.6 mg / kg, for example 4.16 mg / kg; (8) The pharmaceutical composition X is administered according to the weight of the recipient, and the dosage of the PD-1 monoclonal antibody is 0.83-52 mg / kg, for example 0.83 mg / kg; (9) The pharmaceutical composition X includes, but is not limited to, gastrointestinal or injectable administration, preferably oral administration; and (10) The pharmaceutical composition X is prepared as tablets, capsules, granules, powders, pills, powders, suspensions, oral liquids, injections, sprays or drops, preferably tablets, depending on the route of administration.

3. The pharmaceutical composition X according to claim 1 or 2, characterized in that, It meets one or more of the following conditions: (1) In the pharmaceutical composition X, the compound as shown in Formula A or a pharmaceutically acceptable salt thereof is present in an MC solution; the MC solution is an MC solution with a mass concentration of 0.5 g / 100 mL; (2) In the pharmaceutical composition X, the PD-1 monoclonal antibody is present in a physiological saline solution; (3) In the pharmaceutical composition X, the compound of formula A or its pharmaceutically acceptable salt is administered 1-2 times / day, for example once / day; (4) In the pharmaceutical composition X, the PD-1 monoclonal antibody is administered 7-31 days / time or twice / week, for example, 14 days / time; and (5) The mass ratio of the compound of formula A or its pharmaceutically acceptable salt to the PD-1 monoclonal antibody is (2-8):1, for example 5:

1.

4. The pharmaceutical composition X according to claim 3, characterized in that, It meets one or more of the following conditions: (1) In the MC solution, the mass concentration of the compound of formula A or a pharmaceutically acceptable salt thereof is 0.1-50 mg / mL, for example 5 mg / mL; and (2) In the physiological saline solution, the mass concentration of the PD-1 monoclonal antibody is 0.1-20 mg / mL, for example 1 mg / mL.

5. A pharmaceutical composition Y, characterized in that, It includes a first pharmaceutical composition and a second pharmaceutical composition; The first pharmaceutical composition comprises: a compound of formula A or a pharmaceutically acceptable salt thereof and pharmaceutical excipients; The second pharmaceutical composition comprises: a PD-1 monoclonal antibody and pharmaceutical excipients.

6. The pharmaceutical composition Y according to claim 5, characterized in that, It meets one or more of the following conditions: (1) The first drug composition and the second drug composition may be administered simultaneously or separately, for example, separately; (2) The first pharmaceutical composition further includes MC solution; (3) The second pharmaceutical composition further includes physiological saline; (4) The content of the compound of formula A or its pharmaceutically acceptable salt in the first pharmaceutical composition and the content of the PD-1 monoclonal antibody in the second pharmaceutical composition as described in claim 2; (5) The dosage of the drug composition Y is administered according to the weight of the subject, and the dosage of the first drug composition is 25-400 mg / person; and (6) The dosage of the pharmaceutical composition Y is based on the weight of the target, and the dosage of the second pharmaceutical composition is 10-40 mg / kg.

7. The pharmaceutical composition Y according to claim 5 or 6, characterized in that, It meets one or more of the following conditions: (1) In the second pharmaceutical composition, the PD-1 monoclonal antibody is as described in claim 2; (2) The first drug composition is administered 1-2 times / day, for example, once / day; (3) The second drug composition is administered 7-31 days / time or twice / week, for example, 14 days / time; (4) The mass ratio of the compound of formula A or a pharmaceutically acceptable salt thereof in the first pharmaceutical composition to the PD-1 monoclonal antibody in the second pharmaceutical composition as described in claim 3; (5) The first pharmaceutical composition includes, but is not limited to, gastrointestinal or injectable administration, preferably oral administration; (6) The second pharmaceutical composition includes, but is not limited to, gastrointestinal or injectable administration, preferably injectable administration; and (7) The dosage forms of the first pharmaceutical composition and the second pharmaceutical composition are independently tablets, capsules, granules, powders, pills, powders, suspensions, oral liquids, injections, sprays or drops, preferably injection dosage forms.

8. The use of a substance M in the preparation of a medicament for the prevention and / or treatment of tumors; wherein, The substance M is a pharmaceutical composition X as claimed in any one of claims 1-4 or a pharmaceutical composition Y as claimed in any one of claims 5-7.

9. The application as described in claim 8, characterized in that, The tumor in question is a solid tumor; The solid tumor is preferably selected from one or more of breast cancer, gastric cancer, endometrial cancer, cervical cancer, ovarian cancer, and lung cancer, such as breast cancer; And / or, the solid tumor is a locally advanced or metastatic advanced solid tumor with PTEN deficiency or low expression.

10. The application as described in claim 8, characterized in that, The locally advanced or metastatic solid tumors with PTEN deficiency or low expression are selected from one or more of breast cancer, gastric cancer, endometrial cancer, cervical cancer, ovarian cancer, and lung cancer, such as breast cancer.

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