TWI831999B - Chidamide pharmaceutical composition and application thereof - Google Patents

Abstract

The invention relates to the field of pharmaceutical technology and discloses a chidamide pharmaceutical composition and its application. Through research, the present invention finds that one of the reasons for the poor bioavailability of chidamide is that it is a substrate of intestinal P-gp. Therefore, the present invention provides a pharmaceutical combination containing chidamide and a P-glycoprotein inhibitor. The rational application of both substances can significantly improve the bioavailability of chidamide and improve the efficacy of chidamide. Through research, the present invention also found that chidamide combined with P-glycoprotein inhibitors shows good anti-tumor efficacy, and the most significant anti-tumor efficacy is chidamide combined with P-glycoprotein inhibitors and immune checkpoints. inhibitor, indicating that the combined use of chidamide with P-gp inhibitors and immune checkpoint inhibitors can further effectively improve the anti-tumor efficacy of chidamide.

Description

Chidamide pharmaceutical composition and application thereof

This application is based on the application with CN application number 201910804149.8 and the filing date is August 28, 2019, and the CN application number 201911102897.8 and the filing date is November 12, 2019, and claims its priority. The disclosure of the CN application is hereby incorporated into this application in its entirety.

The present invention relates to the field of pharmaceutical technology, and in particular to a chidamide pharmaceutical composition and its application.

Chidamide (English name: Chidamide) is a new anti-cancer drug with a new chemical structure and global intellectual property rights independently designed and synthesized by Shenzhen Microchip Biotechnology Co., Ltd. Chidamide has histone deacetylase inhibitory activity and is an epigenetic regulator drug that can be used to treat a variety of diseases, such as cancer, viral diseases, autoimmune diseases, metabolic diseases, etc. Completed clinical research results show that chidamide has definite effects on various lymphomas, including peripheral T-cell lymphoma, and also on some other solid tumors such as breast cancer, lung cancer, kidney cancer, colorectal cancer, and intrauterine cancer. It has a clinically beneficial effect on patients with membrane cancer, and its comprehensive indicators (safety, preliminary efficacy, pharmacokinetic characteristics) are better than the results of the same period of international studies on drugs with similar mechanisms of action.

After oral solid preparations enter the body, they must go through a dissolution process before they can be absorbed by the body through the biological membrane. Because the dissolution rate of poorly soluble drugs is limited by solubility, which affects drug absorption, they act slowly and have low bioavailability.

Chidamide has very little solubility in water and low bioavailability, which brings about clinical shortcomings such as poor drug absorption, high dosage, and high gastrointestinal toxicity. Therefore, improving the bioavailability of chidamide is of great significance in reducing drug dosage, reducing drug production costs, and reducing gastrointestinal toxicity.

In view of this, the object of the present invention is to provide a chidamide pharmaceutical composition, which can significantly improve the bioavailability of chidamide and can be used in fields such as drug preparation and disease treatment related to chidamide.

In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions: A chidamide pharmaceutical composition includes chidamide and a P-glycoprotein inhibitor, and further includes an immune checkpoint inhibitor.

Chidamide of the present invention has a structure shown in formula (1), and its chemical name is N-(2-amino-4-fluorophenyl)-4-[N-[(E)-3-(3- Pyridine)acrylyl]aminomethyl]benzamide, in its structural formula, the configuration of 3-pyridylacrylyl is E type. Formula 1)

The study of the present invention found that the reason for the low oral bioavailability of chidamide is not only its poor water solubility, but also that chidamide as a substrate of intestinal P-gp is another important reason for reducing its oral bioavailability. The function of the P-gp efflux pump in the digestive tract reduces the absorption of chidamide into the blood. At the same time, the presence of P-gp efflux pumps on the surface of tumor cells can lead to reduced drug efficacy and tumor cell resistance to drugs. Therefore, the present invention starts from discovering the root cause of the problem and provides the combined application of chidamide and P-gp inhibitor, which can significantly improve the bioavailability of chidamide and improve the efficacy of chidamide.

In some embodiments, the pharmaceutical composition includes chidamide and a P-glycoprotein inhibitor. The P-glycoprotein inhibitor of the present invention is selected from the group consisting of synthetic P-glycoprotein inhibitors, natural P-glycoprotein inhibitors and pharmaceutical excipient P-glycoprotein inhibitors. In some embodiments, the P-glycoprotein inhibitor includes, but is not limited to, verapamil, dexverapamil, cyclosporine, dexnicodipine, transfluthixol, tamoxifen, valapamil, PSC833, Tariquidar, TPGS, β-cyclodextrin, PEG, piperine, VX-710, tetrandrine, FG020326, S-9788, GF-120918, LY-335979, DMDCK and MMDCK, optional From any one or two or more thereof; in the specific embodiment of the present invention, the P-glycoprotein inhibitor is preferably verapamil or Tariquidar.

In some embodiments, the mass ratio of chidamide and P-glycoprotein inhibitor is 10:1-1:10; in specific embodiments of the present invention, the mass ratio is 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1: 6, 1:7, 1:8, 1:9 or 1:10. In some embodiments, the mass ratio of chidamide and P-glycoprotein inhibitor is 5:1-1:5, for example, 5:1, 4.5:1, 4:1, 3.5:1, 3: 1, 2.5:1, 2:1, 1.9:1, 1.8:1, 1.7:1, 1.6:1, 1.5:1, 1.4:1, 1.3:1, 1.2:1, 1.1:1, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, 1:2.5, 1:3, 1: 3.5, 1:4, 1:4.5 or 1:5.

In some embodiments, the pharmaceutical composition includes chidamide, and verapamil or Tariquidar, wherein the mass ratio of chidamide to verapamil or Tariquidar is 5:1-1:5, for example 5:1, 4.5:1, 4:1, 3.5:1, 3:1, 2.5:1, 2:1, 1.9:1, 1.8:1, 1.7:1, 1.6:1, 1.5:1, 1.4: 1, 1.3:1, 1.2:1, 1.1:1, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, 1:2.5, 1:3, 1:3.5, 1:4, 1:4.5 or 1:5. In some embodiments, the pharmaceutical composition includes chidamide, a P-glycoprotein inhibitor, and an immune checkpoint inhibitor.

The immune checkpoint inhibitor of the present invention is selected from one or more than two types of anti-CTLA-4 antibodies, anti-PD-1/PD-L1 antibodies, anti-LAG-3 antibodies, anti-TIM-3 antibodies and anti-TIGIT antibodies. Among them, anti-CTLA-4 antibodies include but are not limited to Ipilimumab and Tremelimumab. Anti-PD-1/PD-L1 antibodies include, but are not limited to, RMP 1-14, pembrolizumab, nivolumab, ztezolizumab, avelumab, durvalumab, tislelizumab, camrelizumab, toripalimab, sintilimab, jadecan Nosumab, KN035, GLS-010 and CS1001 can be selected from any one or more than two of them. Anti-LAG-3 antibodies include, but are not limited to, MGD013, FS118, Relatlimab, GSK2831781, LAG525, REGN3767, and TSR033. Anti-TIM-3 antibodies include, but are not limited to, MBG453, TSR022, LY3321367, RO7121661, Sym023, INCAGN2390, and SHR1702. Anti-TIGIT antibodies include, but are not limited to, tiragolumab, BGB-A1217, MK-7684, BMS-986207, AB154, ASP8374, IBI-939 and OMP-313M32.

In some embodiments, the mass ratio of chidamide, P-glycoprotein inhibitor and immune checkpoint inhibitor is (1-10): (1-10): (1-10); wherein, the chidamide Damamide, P-glycoprotein inhibitor and immune checkpoint inhibitor can be independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 within the respective mass ratio range of 1-10 . In a specific embodiment of the present invention, the present invention provides a group of chidamide, P-glycoprotein inhibitors and immune checkpoint inhibitors with a mass ratio of 5:2:2, as an example to demonstrate the drug of the present invention. Anti-tumor effect of the composition.

In the pharmacokinetic test provided by the present invention, the AUC (area under the drug-time curve) of the pharmaceutical composition of the present invention was significantly improved compared to that of chidamide, and the combination multiple was more than 1 times, and the highest was 5 At the same time, the maximum concentration of the drug was also significantly increased, and the combination multiple was greater than 1.5 times, and up to more than 6 times; these results verified that chidamide, as a substrate for intestinal P-gp, reduces its oral bioavailability Another important reason.

In the mouse tumor efficacy test of chidamide combined with P-glycoprotein inhibitors and immune checkpoint inhibitors of the present invention, compared with each single drug test group, the chidamide combined with Tariquidar group showed good anti-tumor effects. The most significant anti-tumor efficacy is the combination of chidamide combined with Tariquidar and Anti-PD-1 antibodies, indicating that the combined use of chidamide with P-gp inhibitors and immune checkpoint inhibitors can further effectively improve Antitumor efficacy of chidamide.

In the present invention's chidamide combined with P-gp inhibitors and immune checkpoint inhibitors enhanced gene expression test of cytokines TNF-α and IL-6, compared with each single drug test group, chidamide combined with Tariquidar The group showed a certain up-regulation of IL-6 and TNF-α gene expression; while the chidamide combined with Tariquidar and Anti-PD-1 antibody group showed a more significant up-regulation of the gene expression of these two cytokines, indicating that The combined use of chidamide with P-gp inhibitors and immune checkpoint inhibitors exerts anti-tumor effects by further promoting the expression of immune cytokines.

Based on the above significant technical advantages, the present invention proposes the use of the pharmaceutical composition in preparing chidamide preparations and/or drugs for treating diseases related to the action mechanism of histone deacetylase.

In some embodiments, diseases related to the histone deacetylase mechanism of action include, but are not limited to, cancer, viral diseases, autoimmune diseases, and hematological diseases. In some embodiments, the disease associated with the histone deacetylase mechanism of action is cancer, such as lymphoma, breast cancer, lung cancer, renal cancer, colorectal cancer, or endometrial cancer. In some embodiments, the disease associated with the histone deacetylase mechanism of action is peripheral T-cell lymphoma.

Based on the above applications, the present invention also provides a chidamide preparation, which includes the chidamide pharmaceutical composition of the present invention, and more specifically includes the chidamide pharmaceutical composition and pharmaceutically acceptable excipients. Among them, the dosage forms of this preparation include but are not limited to tablets, capsules, pills, oral liquid preparations (such as syrups), granules, powders, ointments and dropping pills. All preparations can be prepared as sustained-release pharmaceuticals or controlled-release pharmaceuticals. Any pharmaceutical release form.

The pharmaceutically acceptable excipients can be adjusted and selected according to the dosage form to be prepared. In the present invention, the auxiliary material may be selected from one or more of fillers, disintegrants, binders, and lubricants.

On the basis of the above, the present invention further provides a pharmaceutical combination including chidamide and a P-glycoprotein inhibitor. In some embodiments, the P-glycoprotein inhibitor is selected from the group consisting of synthetic P-glycoprotein inhibitors, natural P-glycoprotein inhibitors and pharmaceutical excipient P-glycoprotein inhibitors. In some embodiments, the P-glycoprotein inhibitor includes, but is not limited to, verapamil, dexverapamil, cyclosporine, dexnicodipine, transfluthixol, tamoxifen, valapamil, Any of PSC833, Tariquidar, TPGS, β-cyclodextrin, PEG, piperine, VX-710, tetrandrine, FG020326, S-9788, GF-120918, LY-335979, DMDCK and MMDCK One or more than two types. In a specific embodiment of the present invention, the P-glycoprotein inhibitor is preferably verapamil or Tariquidar.

In some embodiments, the mass ratio of chidamide and P-glycoprotein inhibitor is 10:1-1:10; in specific embodiments of the present invention, the mass ratio is 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1: 6, 1:7, 1:8, 1:9 or 1:10. In some embodiments, the mass ratio of chidamide and P-glycoprotein inhibitor is 5:1-1:5, for example, 5:1, 4.5:1, 4:1, 3.5:1, 3: 1, 2.5:1, 2:1, 1.9:1, 1.8:1, 1.7:1, 1.6:1, 1.5:1, 1.4:1, 1.3:1, 1.2:1, 1.1:1, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, 1:2.5, 1:3, 1: 3.5, 1:4, 1:4.5 or 1:5.

In some embodiments, the drug combination includes chidamide, and verapamil or Tariquidar, wherein the mass ratio of chidamide to verapamil or Tariquidar is 5:1-1:5, for example, 5 :1, 4.5:1, 4:1, 3.5:1, 3:1, 2.5:1, 2:1, 1.9:1, 1.8:1, 1.7:1, 1.6:1, 1.5:1, 1.4:1 , 1.3:1, 1.2:1, 1.1:1, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1 :1.9, 1:2, 1:2.5, 1:3, 1:3.5, 1:4, 1:4.5 or 1:5.

In some embodiments, the drug combination includes chidamide, a P-glycoprotein inhibitor, and an immune checkpoint inhibitor. In some embodiments, the immune checkpoint inhibitor is selected from one or both of anti-CTLA-4 antibodies, anti-PD-1/PD-L1 antibodies, anti-LAG-3 antibodies, anti-TIM-3 antibodies, and anti-TIGIT antibodies. More than one species. In some embodiments, the anti-CTLA-4 antibodies include, but are not limited to, Ipilimumab and Tremelimumab. In some embodiments, the anti-PD-1/PD-L1 antibodies include, but are not limited to, RMP 1-14, pembrolizumab, nivolumab, ztezolizumab, avelumab, durvalumab, tislelizumab, camrelizumab, toripalimab , sintilimab, jenosumab, KN035, GLS-010 and CS1001, you can choose from any one or more than two of them. In some embodiments, the anti-LAG-3 antibodies include, but are not limited to, MGD013, FS118, Relatlimab, GSK2831781, LAG525, REGN3767, and TSR033. Anti-TIM-3 antibodies include, but are not limited to, MBG453, TSR022, LY3321367, RO7121661, Sym023, INCAGN2390, and SHR1702. In some embodiments, the anti-TIGIT antibodies include, but are not limited to, tiragolumab, BGB-A1217, MK-7684, BMS-986207, AB154, ASP8374, IBI-939, and OMP-313M32.

In some embodiments, the mass ratio of chidamide, P-glycoprotein inhibitor and immune checkpoint inhibitor is (1-10): (1-10): (1-10); wherein, the chidamide Damamide, P-glycoprotein inhibitor and immune checkpoint inhibitor can be independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 within the respective mass ratio range of 1-10 .

In some embodiments, the chidamide, P-glycoprotein inhibitor, and immune checkpoint inhibitor are in the same or separate formulation units.

In another aspect, the invention provides a method of inhibiting HDAC, which comprises administering to a subject in need thereof an effective amount of a chidamide pharmaceutical composition, chidamide preparation or pharmaceutical combination described herein. steps.

In some embodiments, the drugs in the pharmaceutical composition can be administered simultaneously, separately, or sequentially.

In another aspect, the present invention provides a method for treating diseases related to the mechanism of action of histone deacetylase, comprising administering to a subject in need thereof an effective amount of a chidamide pharmaceutical composition described herein, Procedures for Chidamide Preparation or Drug Combination.

In some embodiments, the disease associated with the histone deacetylase mechanism of action is selected from the group consisting of cancer, viral diseases, autoimmune diseases, and hematological diseases. In some embodiments, the disease associated with the histone deacetylase mechanism of action is cancer, such as lymphoma, breast cancer, lung cancer, kidney cancer, colorectal cancer, or endometrial cancer. In some embodiments, the disease associated with the histone deacetylase mechanism of action is peripheral T-cell lymphoma.

In some embodiments, the drugs in the pharmaceutical composition can be administered simultaneously, separately, or sequentially.

As can be seen from the above technical solution, the present invention has discovered through research that one of the reasons for the poor bioavailability of chidamide is that it is a substrate of intestinal P-gp. Therefore, the present invention provides a product containing chidamide and P-glycoside. The rational application of protein inhibitor pharmaceutical compositions can significantly improve the bioavailability of chidamide and improve the efficacy of chidamide. Through research, the present invention also found that chidamide combined with a P-glycoprotein inhibitor shows good anti-tumor efficacy, and the most significant anti-tumor efficacy is chidamide combined with a P-glycoprotein inhibitor and an immune checkpoint inhibitor, indicating that the combined use of chidamide with P-gp inhibitors and immune checkpoint inhibitors can further effectively improve the anti-tumor efficacy of chidamide.

The invention discloses a chidamide pharmaceutical composition and its preparation method and application. Persons skilled in the art can learn from the content of this article and appropriately improve the process parameters to achieve it. It should be noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are deemed to be included in the present invention. The pharmaceutical compositions and their applications of the present invention have been described through preferred embodiments. Relevant personnel can obviously make modifications or appropriate changes to the pharmaceutical compositions and their applications herein without departing from the content, spirit and scope of the present invention. and combination to implement and apply the technology of the present invention.

The chidamide pharmaceutical composition and its application provided by the present invention will be further described below. Example 1: Absorption experiment of Caco-2 cells cultured in vitro

Cell culture: Coca-2 cells were seeded on polyethylene membrane (PET) in a 96-well plate insert system at 2 x ¹⁰ cells per square centimeter until day 21-28 to form a confluent cell monolayer. Change the culture medium every 3-4 days.

Experimental method: Dilute the test compound to a concentration of 10 μM in transport buffer (HBSS + 1% BSA) in 10 mM stock solution and apply it to the apical or basolateral side of the cell monolayer. The penetration of the test compound from A to B or B to A was determined repeatedly during a 120-minute incubation at 37°C and 5% carbon dioxide at a relative humidity of 95%. In addition, the efflux ratio of each compound was determined. Test and reference compounds were quantified using LC-MS/MS analysis based on the analyte/Is peak area ratio. The experimental results are as follows in Table 1: Table 1 Summary Mean Papp (10 ^-6 cm/s) Efflux Ratio Rank Compound ID A to B B to A Papp P-gp Atenolol 1.73 1.33 0.77 Low Propranolol 22.10 11.16 0.51 High Digoxin 0.48 8.06 16.64 Low Substrate Digoxin+GF120918A 3.83 3.16 0.82 Moderate CS055 0.96 3.24 3.38 Low Substrate CS055+GF120918A 1.45 1.47 1.01 Low CS02 0.84 0.94 1.12 Low CS02+GF120918A 0.81 0.69 0.85 Low CS03 8.22 4.85 0.59 Moderate CS03+GF120918A 7.89 3.01 0.38 Moderate CS04 2.91 5.16 1.78 Moderate CS04+GF120918A 5.11 3.69 0.72 Moderate CS055 in the table above is chidamide, and the efflux ratio (Efflux ratio) is calculated from Papp(B to A)/Papp(A to B). When the ratio is greater than 2, it indicates that the efflux is mediated by P-gp. The platoon mechanism works. The efflux rate of chidamide reached 3.38, indicating that chidamide is a substrate of P-gp. Except for Digoxin, the ratios of other drugs are less than 2, indicating that not all drugs are substrates of P-glycoprotein. Example 2: Chidamide combined with verapamil pharmacokinetics test in rats

Twelve SD rats were selected and randomly divided into 2 groups, with 6 rats in each group, half male and half female. During the test, SD rats were fasted overnight before administration. Each rat in group 1 was orally administered chidamide suspension at 20 mg/kg alone, and each rat in group 2 was orally administered a mixture of chidamide (20 mg/kg) and verapamil (25 mg/kg) at the same time. suspension. Blood samples were collected before administration and 15min, 0.5h, 1h, 2h, 4h, 6h, and 8h after administration. Each sample was collected about 0.2ml. Heparin sodium was anticoagulated. After collection, place it on ice and centrifuge within 1 hour. The plasma was separated, the drug was extracted with acetonitrile, and the drug concentration in the plasma was measured using LC-MS/MS. The test results are shown in Table 2 and Figure 1. Table 2 Comparison of pharmacokinetic parameters of two groups of animals Pharmacokinetic parameters unit chidamide Chidamide + Verapamil combination multiple AUC _(0-t) h·ng/mL 1688.1 4789.4 2.83 C _max ng/mL 703.6 2765.6 3.93 Pharmacokinetic experiments in rats showed that when chidamide was combined with the first-generation P-gp inhibitor verapamil, the plasma exposure AUC increased to 2.83 times and C _max increased to 3.93 times. The above experiments further confirmed that chidamide is a substrate of P-gp, and P-gp inhibitors can significantly increase the in vivo bioavailability of chidamide. Example 3: Chidamide combined with Tariquidar pharmacokinetics test in rats

Twelve SD rats were selected and randomly divided into 2 groups, with 6 rats in each group, half male and half female. During the test, SD rats were fasted overnight before administration. Each rat in group 1 was orally administered chidamide suspension at 20 mg/kg alone, and each rat in group 2 was orally administered chidamide (20 mg/kg) and Tariquidar (12 mg/kg) suspension at the same time. Blood samples were collected before administration and 15min, 0.5h, 1h, 2h, 4h, 6h, and 8h after administration. Each sample was collected about 0.2ml. Heparin sodium was anticoagulated. After collection, place it on ice and centrifuge within 1 hour. The plasma was separated, the drug was extracted with acetonitrile, and the drug concentration in the plasma was measured using LC-MS/MS. The test results are shown in Table 3 and Figure 2. Table 3 Comparison of pharmacokinetic parameters of two groups of animals Pharmacokinetic parameters unit Chidamide Chidamide + Tariquidar combination multiple AUC _(0-t) h·ng/mL 2757.1 14517.6 5.26 C _max ng/mL 970.0 6139.6 6.33 Pharmacokinetic experiments in rats showed that when chidamide was combined with the third-generation P-gp inhibitor Tariquidar, the plasma exposure AUC increased significantly to 5.26 times, and the C _max increased significantly to 6.33 times. The above experiments further confirmed that chidamide is a substrate of P-gp, and P-gp inhibitors can significantly increase the in vivo bioavailability of chidamide. Example 4: Chidamide combined with vitamin E polyethylene glycol succinate (TPGS) pharmacokinetic test in rats

Twelve SD rats were selected and randomly divided into 2 groups, with 6 rats in each group, half male and half female. During the test, SD rats were fasted overnight before administration. Each rat in group 1 was orally administered chidamide suspension at 20 mg/kg, and each rat in group 2 was orally administered chidamide (20 mg/kg) and TPGS (3.3 mg/kg) suspension at the same time. . Blood samples were collected before administration and 15min, 0.5h, 1h, 2h, 4h, 6h, and 8h after administration. Each sample was collected about 0.2ml. Heparin sodium was anticoagulated. After collection, place it on ice and centrifuge within 1 hour. The plasma was separated, the drug was extracted with acetonitrile, and the drug concentration in the plasma was measured using LC-MS/MS. The test results are shown in Table 4 and Figure 3. Table 4 Comparison of pharmacokinetic parameters of two groups of animals Pharmacokinetic parameters unit Chidamide Chidamide+TPGS combination multiple AUC _(0-t) h·ng/mL 2757.1 2782.4 1 C _max ng/mL 970.0 1685.1 1.73 Pharmacokinetic experiments in rats show that when chidamide is combined with the pharmaceutical excipient P-gp inhibitor TPGS, the plasma exposure AUC does not change much, but the C _max can be increased to 1.73 times that of the drug alone. . It shows that the pharmaceutical excipient P-gp inhibitor can also promote the absorption of chidamide. However, the effect is less than that of P-gp inhibitors such as verapamil and Tariquidar. Example 5: Rat pharmacokinetic test of chidamide combined with hydroxypropyl β-cyclodextrin

Twelve SD rats were selected and randomly divided into 2 groups, with 6 rats in each group, half male and half female. During the test, SD rats were fasted overnight before administration. Each rat in group 1 was orally administered chidamide suspension at 20 mg/kg alone, and each rat in group 2 was orally administered chidamide (20 mg/kg) mixed with 40% hydroxypropyl β-cyclodextrin. suspension. Blood samples were collected before administration and 15min, 0.5h, 1h, 2h, 4h, 6h, and 8h after administration. Each sample was collected about 0.2ml. Heparin sodium was anticoagulated. After collection, place it on ice and centrifuge within 1 hour. The plasma was separated, the drug was extracted with acetonitrile, and the drug concentration in the plasma was measured using LC-MS/MS. The test results are shown in Table 5 and Figure 4. Table 5 Comparison of pharmacokinetic parameters of two groups of animals Pharmacokinetic parameters unit Chidamide Chidamide + Hydroxypropyl β-Cyclodextrin combination multiple AUC _(0-t) h·ng/mL 2757.1 3020.6 1.1 _Cmax ng/mL 970.0 1593.0 1.64 The results of pharmacokinetic experiments in rats show that when chidamide is combined with the pharmaceutical excipient P-gp inhibitor hydroxypropyl β-cyclodextrin, the plasma exposure AUC increases to 1.1 times, and the C _max increases to 1.64 times that of medication alone. It shows that pharmaceutical excipient P-gp inhibitors can also promote the absorption of chidamide, but the effect is insufficient compared to P-gp inhibitors such as verapamil and Tariquidar. Example 6: Mouse tumor efficacy test of chidamide combined with P-gp inhibitor

Forty female BALB/c mice aged 6-7 weeks were selected and randomly divided into 4 groups, with 10 mice in each group. At the beginning of the experiment, each mouse was subcutaneously inoculated with mouse intestinal cancer cell CT26 in the armpit, 5*10 ⁵ cells/mouse. Administration was started ⁷ days after inoculation when the tumor grew to 50mm3, and single-agent chidamide (25 mg/kg), chidamide combined with Tariquida, chidamide combined with verapamil, and chidamide were administered respectively. Amine combined with TPGS was administered continuously until 21 days after vaccination, during which the tumor growth of each group was measured every two days. The dosage and test results of each group are shown in Figure 5.

As can be seen from Figure 5, among the various test groups, the chidamide combined with Tariquidar group has the best anti-tumor efficacy, followed by the chidamide combined with verapamil group and the chidamide combined with TPGS group. The anti-tumor efficacy of each combination group is The efficacy was better than that of chidamide alone, indicating that the combined use of chidamide and P-gp inhibitors can effectively improve the anti-tumor efficacy of chidamide. Example 7: Mouse tumor efficacy test of chidamide combined with P-gp inhibitor and immune checkpoint inhibitor

Sixty female BALB/c mice aged 6-7 weeks were selected and randomly divided into 6 groups, with 10 mice in each group. At the beginning of the experiment, each mouse was subcutaneously inoculated with mouse intestinal cancer cell CT26 in the armpit, 5*10 ⁵ cells/mouse. Administration was started ⁷ days after the inoculation when the tumor grew to 50mm3. The solvent control, single drug chidamide (25 mg/kg, gavage, once a day), and single drug Tariquidar (10 mg/kg, gavage, once a day) were given respectively. once), single-agent Anti-PD-1 antibody (RMP 1-14, BioXcell, 10 mg/kg, intraperitoneal injection, twice a week) and chidamide (25 mg/kg, gavage, once a day) combined with Tariquidar (10 mg /kg, gavage, once a day), and chidamide (25 mg/kg, gavage, once a day) combined with Tariquidar (10 mg/kg, gavage, once a day) and Anti-PD-1 antibody (RMP 1- 14, BioXcell, 10 mg/kg, intraperitoneal injection, twice a week), administered continuously for 15 days, during which the tumor growth of each group was measured every two days. The dosage and test results of each group are shown in Figure 6.

As can be seen from Figure 6, compared with each single drug test group, the chidamide combined with Tariquidar group showed good anti-tumor efficacy; and the most significant anti-tumor efficacy was achieved with chidamide combined with Tariquidar and Anti-PD-1 antibody group, indicating that the combined use of chidamide with P-gp inhibitors and immune checkpoint inhibitors can further effectively improve the anti-tumor efficacy of chidamide. Example 8: Chidamide combined with P-gp inhibitor and immune checkpoint inhibitor enhances gene expression of cytokines TNF-α and IL-6

At the end of the test in Example 7 above, the spleens of the mice in each test group were taken. First, a single spleen cell suspension is obtained through steps such as separation and removal of red blood cells. 5*10 ⁶ spleen cells were taken from each group, and total RNA was extracted using TRIzol reagent (Invitrogen Company). cDNA was obtained by reverse transcription using the Transcriptor First Strand cDNA Synthesis Kit (Roche), and then real-time quantitative PCR was performed in an ABI Prism 7000 PCR instrument using Roche's SYBR Green Master Dye and other reagents. The primer sequences of mouse cytokine IL-6 and TNF-α genes are respectively, IL-6 Forward: 5'-GGAGCCCACCAAGAACGATAG-3', IL-6 Reverse: 5'-GTGAAGTAGGG AAGGCCGTG-3'; TNF-α Forward: 5'-TGATCGGTCCCCAAAGGGAT-3', TNF-α Reverse: 5'-GCTACGACGTGGGCTACAGG-3'. The primer sequences of mouse internal control β-actin are: β-actin Forward: 5'-GTCGAGTCGCGTCCACC-3', β-actin Reverse: 5'-ACGATGGAGGGGAATACAGC-3'. The relative expression results of spleen cytokines IL-6 and TNF-α genes in each group are shown in Figure 7.

As can be seen from Figure 7, compared with each single drug test group, the chidamide combined with Tariquidar group showed a certain effect in up-regulating the expression of IL-6 and TNF-α genes; while the chidamide combined with Tariquidar and Anti-PD-1 antibody The group showed a more significant up-regulation of the gene expression of these two cytokines, indicating that the combined use of chidamide with P-gp inhibitors and immune checkpoint inhibitors exerts anti-tumor effects by further promoting the expression of immune cytokines. effect.

The above are only the preferred embodiments of the present invention. It should be pointed out that those of ordinary skill in the art can also make several improvements and modifications without departing from the principles of the present invention. These improvements and modifications It should also be regarded as the protection scope of the present invention.

without.

Figure 1 shows the results of the pharmacokinetics experiment of chidamide combined with verapamil in rats; Figure 2 shows the results of pharmacokinetic experiments on chidamide combined with Tariquidar in rats; Figure 3 shows the results of pharmacokinetic experiments on chidamide combined with TPGS in rats; Figure 4 shows the results of pharmacokinetic experiments on chidamide combined with hydroxypropyl β-cyclodextrin in rats; Figure 5 shows the mouse tumor efficacy test of chidamide combined with a P-gp inhibitor; Figure 6 shows the mouse tumor efficacy test of chidamide combined with P-gp inhibitors and immune checkpoint inhibitors; Figure 7 shows that chidamide combined with P-gp inhibitors and immune checkpoint inhibitors enhances the gene expression of cytokines TNF-α and IL-6.

Claims (8)

A chidamide pharmaceutical composition is characterized by including chidamide and a P-glycoprotein inhibitor, and the P-glycoprotein inhibitor is Tariquidar. The pharmaceutical composition according to claim 1, wherein the mass ratio of chidamide and P-glycoprotein inhibitor is 10:1-1:10. The pharmaceutical composition according to claim 1 or claim 2, which further includes an anti-PD-1/PD-L1 antibody, wherein the anti-PD-1/PD-L1 antibody is selected from RMP 1-14, pembrolizumab pembrolizumab, nivolumab, atezolizumab, avelumab, durvalumab, tislelizumab ), camrelizumab, toripalimab, sintilimab, jenosumab, envolizumab (KN035), cepalizumab (GLS-010) and sugalizumab (CS1001) one or more than two. The pharmaceutical composition according to claim 3, wherein the mass ratio of chidamide, P-glycoprotein inhibitor and anti-PD-1/PD-L1 antibody is (1-10): (1-10): (1-10). The use of a pharmaceutical composition as described in any one of claims 1 to 4 in preparing chidamide preparations and/or medicaments for treating lymphoma, breast cancer, lung cancer, kidney cancer or colorectal cancer. The use as described in claim 5, wherein the lymphoma is peripheral T-cell lymphoma. A chidamide preparation, characterized in that it contains the chidamide pharmaceutical composition described in any one of claims 1 to 4. The preparation according to claim 7, wherein the dosage forms of the preparation include tablets, capsules, pills, oral liquid preparations (syrups), granules, powders, ointments and dropping pills.