WO2022134433A1

WO2022134433A1 - Composition and application thereof in treatment of tumors

Info

Publication number: WO2022134433A1
Application number: PCT/CN2021/093820
Authority: WO
Inventors: 朱永亮; 赵志浩; 张宝钰; 胡代强; 刘丹; 张叶静
Original assignee: 苏州普瑞森基因科技有限公司
Priority date: 2020-12-21
Filing date: 2021-05-14
Publication date: 2022-06-30
Also published as: CN114642732A

Abstract

The present invention relates to the technical field of pharmaceuticals, and relates in particular to a composition and an application thereof in the treatment of tumors. The composition consists of baicalein and a PD-1 antagonist. Experiments show that the anti-tumor efficacy of baicalein combined with PD-1 is significantly higher than that of each of the two components alone, indicating that the combination of the two has a synergistic effect in fighting tumors.

Description

A kind of composition and its application in the treatment of tumor

This application claims the priority of the Chinese patent application filed on December 21, 2020 with the application number 202011518837.7 and the invention titled "a composition and its application in the preparation of a drug for the treatment of tumors", all of which are The contents are incorporated herein by reference.

technical field

The present invention relates to the technical field of medicine, in particular to a composition and its application in the treatment of tumors.

Background technique

Colon cancer is a common malignant tumor of the digestive tract that occurs in the colon, and its incidence is increasing year by year in my country. Colon cancer is mainly adenocarcinoma, mucinous adenocarcinoma, undifferentiated carcinoma. The general shape was polypoid, ulcerated and so on. Colon cancer can develop in a circle along the intestinal wall, spread up and down along the longitudinal diameter of the intestinal canal, or infiltrate deep into the intestinal wall. In addition to lymphatic vessels, blood flow metastasis and local invasion, it can also be implanted in the abdominal cavity or spread along the suture and incision surface. . Patients with chronic colitis, colon polyps, and male obesity are susceptible groups.

Currently, the main treatments for colon cancer are surgery, radiotherapy and chemotherapy. The choice of treatment depends on the location of the tumor, the degree of malignancy, the degree of development, and the patient's physical state. Although the effect of surgical resection is the best, the risk of surgery is high and the recurrence rate after surgery is high, especially in patients with advanced disease who are not suitable for surgical treatment. The side effects of radiotherapy and chemotherapy drugs. Immunotherapy is another effective treatment method in addition to surgery and chemotherapy, and it has attracted more and more attention of researchers. Among them, PD-1 (programmed death receptor 1) is recognized to play an important role in immune regulation and peripheral tolerance maintenance. PD-1 (programmed death receptor 1) is modestly expressed on naive T, B and NKT cells and is upregulated by T/B cell receptor signaling on lymphocytes, monocytes and myeloid cells. However, PD-1 is considered important in immune regulation and maintenance of peripheral tolerance, and monoclonal antibodies that inhibit PD-1 are in clinical development for cancer therapy. Currently, PD-1 antagonists are only In the course of treatment of some patients, it showed better therapeutic effect. Many patients have low response rates to PD-1 antagonists. Therefore, it is of great significance to provide a composition with a significant therapeutic effect on colon cancer.

SUMMARY OF THE INVENTION

In view of this, the present invention provides a composition and its application in the treatment of tumors. The combination of baicalein and PD-1 in the composition has an obvious inhibitory effect on the growth of colon cancer tumor volume, and the effect is significantly better than that of the two components alone.

In order to achieve the above-mentioned purpose of the invention, the present invention provides the following technical solutions:

The present invention provides a composition consisting of baicalein and a PD-1 antagonist.

PD-1 is recognized to play an important role in immune regulation and maintenance of peripheral tolerance. PD-1 is modestly expressed on naive T, B and NKT cells and is upregulated by T/B cell receptor signaling on lymphocytes, monocytes and myeloid cells. The PD1 inhibitor is an antibody protein against PD1, blocking this pathway, the two proteins cannot combine, so that T cells can continue to kill tumor cells. Taking this as the target endows immune cells with stronger killing activity and realizes the precise treatment of tumors.

Baicalein is a pleiotropic natural active substance whose structural formula is shown in formula I. Studies have shown that baicalein can inhibit the proliferation of various tumor cells, specifically induce tumor cell apoptosis, block tumor cell cycle, inhibit telomerase activity, and tumor angiogenesis, invasion, and metastasis. Variety and less toxic and side effects. However, it is less effective in the treatment of colon cancer.

The present invention combines two substances, baicalein and PD-1 (programmed death receptor 1) antagonist, which have completely different action mechanisms and action pathways. The response rate showed significant anti-tumor efficacy, and the efficacy was more obvious in patients with tumors that were not sensitive to PD-1 antagonists.

In some embodiments, the mass ratio of the baicalein and the PD-1 antagonist is (25-150): (5-10).

In some specific embodiments, the mass ratio of baicalein and PD-1 antagonist is 25:5.

The present invention also provides the application of the composition in the preparation of a medicament for inhibiting the growth of tumor volume.

Specifically, the inhibiting tumor volume growth includes inhibiting tumor cell growth and/or proliferation.

The present invention also provides the application of the composition in the treatment of tumors. Meanwhile, the present invention also provides the application of baicalein in enhancing the response rate of tumor patients to PD-1 antagonist and the application of baicalein and PD-1 antagonist in treating tumor.

Wherein, the treating tumor includes inhibiting tumor volume growth and/or inhibiting tumor cell migration.

In the present invention, treating tumors and inhibiting the growth of tumor volume means that after the use of baicalein and PD-1 antagonists on tumor patients, there are obvious therapeutic effects, such as inhibition of tumor volume growth and inhibition of tumor cell migration.

Wherein, the tumor includes but is not limited to gastric cancer, intestinal cancer, melanoma, breast cancer, esophageal cancer, lung cancer, ovarian cancer, cervical cancer, kidney cancer, bladder cancer, pancreatic cancer, prostate cancer, non-Hodge cancer Gold lymphoma, chronic lymphoma leukemia, multiple myeloma, acute myeloid leukemia, or acute lymphoma leukemia.

In some embodiments, the tumor is a solid tumor, including but not limited to gastric cancer, bowel cancer, melanoma, breast cancer, esophageal cancer, lung cancer, ovarian cancer, cervical cancer, kidney cancer, bladder cancer, pancreatic cancer, prostate cancer, Non-Hodgkin's Lymphoma. In some preferred embodiments, the tumor is a PD-L expressing cancer. In some more preferred embodiments, the tumor includes colon cancer, melanoma, and triple negative breast cancer. In some specific embodiments, the tumor is colon cancer.

Specifically, baicalein is used in combination with PD-1 antagonists to treat tumors, and the therapeutic effect on tumors is achieved by enhancing the response rate of tumor patients to PD-1 antagonists through baicalein, thereby inhibiting the growth of tumor volume , the migration of tumor cells, so as to play a role in the treatment of tumors.

In the scheme of using baicalein and a PD-1 antagonist to treat tumors in combination according to the present invention, baicalein and a PD-1 antagonist are administered to tumor patients, preferably tumor patients with positive PD-L1 expression. The combination therapy of the present invention can be used for patients who have not received biological therapy or chemotherapy, that is, the first treatment patients; and can also be used for patients who have undergone biological therapy or chemotherapy but have no obvious therapeutic effect.

In the course of combination therapy, the administration of baicalein and PD-1 antagonist includes various forms, which can be administered orally or parenterally, such as intravenous, subcutaneous, intramuscular, subperitoneal, rectal, or transdermal routes.

The present invention also provides a medicament for treating tumors comprising the composition of the present invention.

In the compositions, medicines and applications of the present invention, baicalein can be obtained commercially, and can also be prepared by common methods of extracting baicalein in the art, such as ethanol extraction, ethanol ultrasonic extraction and other methods. The existing form of baicalein is not particularly limited, and it can be in the form of a compound of baicalein or in the form of a preparation of baicalein. When the existing form is a preparation, the dosage form of the preparation is an oral preparation, such as tablet, capsule, pill, granule, decoction, ointment, lotion, oral liquid, drop pill or syrup and the like.

In a specific embodiment of the present invention, baicalein is prepared by ethanol flash extraction. Specific steps are as follows:

Weigh 50 g of Scutellaria baicalensis coarse powder, add it into warm water for self-enzymatic hydrolysis for three hours, add 10 times the amount of 90% ethanol to Scutellaria baicalensis after enzymatic hydrolysis, flash extract twice for 90 s each time, and filter the extract. Scutellaria baicalensis extract was recovered by part of ethanol to 500ml, and 10ml of 1% chitosan acetic acid solution was added to each 100ml of medicinal solution for adsorption and clarification. , the content of baicalein in the product was measured to be 78.1%.

In the above medicines and applications of the present invention, the PD-1 antagonist is a PD-1 monoclonal antibody or an antigen-binding fragment thereof. Wherein, the PD-1 monoclonal antibody or its antigen-binding fragment specifically binds to PD-1 and blocks the combination of PD-1 and PD-L1. Preferably, the PD-1 monoclonal antibody or its antigen-binding fragment specifically binds to human PD-1 and blocks the binding of human PD-1 and human PD-L1.

The present invention has no special limitation on the source of the PD-1 antagonist, which can be purchased from any manufacturer. There is no special restriction on its dosage form, including but not limited to tablets, capsules, pills, granules, decoctions, ointments, lotions, oral liquids, injections, drop pills or syrups, preferably injections, such as injection powder and Injections.

In the scheme of using baicalein and PD-1 antagonist for combined treatment of tumors according to the present invention, each preparation can be administered simultaneously or sequentially in any order. The specific administration dose can be determined according to the specific condition of the tumor patient combined with the clinical experience of the doctor. In some embodiments, the effective dose of baicalein is 25-150 mg/kg, and the effective dose of PD-1 antagonist is 5-10 mg/kg. In some specific embodiments of the present invention, the effective dose of baicalein is 100 mg/kg, and the effective dose of PD-1 antagonist is 5 mg/kg.

In the compositions, medicines and applications of the present invention, the tumors include but are not limited to gastric cancer, intestinal cancer, melanoma, breast cancer, esophageal cancer, lung cancer, ovarian cancer, cervical cancer, kidney cancer, and bladder cancer , pancreatic cancer, prostate cancer, non-Hodgkin lymphoma, chronic lymphoma leukemia, multiple myeloma, acute myeloid leukemia or acute lymphoma leukemia.

In the composition, medicine and application of the present invention, the tumor is a solid tumor, including but not limited to gastric cancer, intestinal cancer, melanoma, breast cancer, esophageal cancer, lung cancer, ovarian cancer, cervical cancer, kidney cancer, and bladder cancer , pancreatic cancer, prostate cancer, non-Hodgkin lymphoma.

In some preferred embodiments, the tumor is a PD-L expressing cancer. In some more preferred embodiments, the tumor includes colon cancer, melanoma, and triple negative breast cancer. In some specific embodiments, the tumor is colon cancer.

Experiments show that the combination of baicalein and PD-1 in the present invention is used for the treatment of tumors, and the curative effect is significantly higher than that of the two components administered alone, indicating that the combination of the two produces a significant synergistic effect in anti-tumor.

Description of drawings

Figure 1 shows the changes in the body weight of mice in each group after administration;

Figure 2 shows the tumor volume and relative tumor inhibition rate of mice in each group after administration.

Detailed ways

The present invention provides a composition and its application in the treatment of tumors, and those skilled in the art can learn from the content of this article and appropriately improve the process parameters to achieve. It should be particularly pointed out 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 method and application of the present invention have been described through the preferred embodiments, and it is obvious that relevant persons can make changes or appropriate changes and combinations of the methods and applications herein without departing from the content, spirit and scope of the present invention to realize and apply the present invention. Invention technology.

The test materials used in the present invention are all common commercial products and can be purchased in the market.

Below in conjunction with embodiment, the present invention is further elaborated:

Example 1 Antitumor effect of the composition of the present invention in the CT-26 model of murine colon cancer

Cell culture: CT-26 cells were cultured in RPMI1640 medium containing 10% fetal bovine serum. CT-26 cells in exponential growth phase were collected and resuspended in PBS to an appropriate concentration for subcutaneous tumor inoculation in mice.

Tumor cell inoculation: experimental mice were subcutaneously inoculated with 4×10 ⁷ CT-26 cells on the right back, and the cells were resuspended in PBS (0.1 ml/mice), and the tumor growth was observed regularly.

Randomization: Before dosing began, all animals were weighed and tumor volumes were measured with vernier calipers. Since the tumor volume will affect the effectiveness of the treatment, a randomization design was used to group the mice according to their tumor volume to ensure that the tumor volume between the different groups is similar.

When tumors grew to an average volume of about 100 mm ³ , they were randomized into groups based on tumor size and mouse body weight. The experiment was divided into test group (group B), positive control group (group P), combined medication group (group BP) and vehicle control group (group V).

Administration: The drugs involved in the present invention include baicalein and PD-1 inhibitors.

Wherein, the preparation method of baicalein is as follows: Weigh 50 g of scutellaria baicalensis coarse powder, add it into warm water for self-enzymatic hydrolysis for three hours, add 10 times the amount of 90% ethanol after enzymatic hydrolysis, and perform flash extraction twice, 90s each time, and the extract is filtered. Scutellaria baicalensis extract recovers part of ethanol to 500ml, adds 10ml of 1% chitosan acetic acid solution per 100ml of medicinal solution for adsorption and clarification, continues to concentrate the clarified medicinal solution to 100ml, adds hydrochloric acid to the concentrated medicinal solution to make it acidic, and centrifuge to obtain 4.746g of dry product , the content of baicalein in the product was measured to be 78.1%.

PD-1 antagonist was purchased from BioXcell, InVivoMAb anti-mouse PD-1 (CD279).

The specific administration method, dosage and route of administration are shown in Table 1. The day of tumor cell inoculation was defined as day 0.

Table 1 Administration route, dose and schedule in CT-26 animal model

The curative effect was evaluated according to the relative tumor inhibition rate and tumor delay time, and the drug efficacy was evaluated according to the changes of animal body weight and death. The main observation indicators include:

Relative tumor inhibition rate TGI (%): TGI%=(1-T/C)×100%. T/C% is the relative tumor proliferation rate, the percentage value of the relative tumor volume or tumor weight of the treatment group and the control group at a certain time point.

Tumor size calculation formula: tumor volume (mm ³ )=0.5×(tumor long diameter×tumor short diameter ² ).

Routine monitoring: activity of laboratory animals, food and water intake, weight gain or loss, eyes, coat and other abnormalities.

The results showed that there was no animal death in each treatment group, no obvious drug toxicity, and no significant change in the body weight of the mice. The results are shown in Figure 1. The tumor growth of each treatment group and control group is shown in Table 2 and Figure 2.

Table 2. Changes in tumor volume of mice in each group with treatment time in CT-26 mouse colon cancer model

Results and discussion:

20 days after administration, the combination group of baicalein (100mg/kg) and PD-1 (5mg/kg) of the present invention has a significant inhibitory effect on the growth of tumor volume, and there is a very significant difference compared with the vehicle control group (p <0.001). Baicalein (100mg/kg), baicalein (105mg/kg), PD-1 antagonist (5mg/kg) and PD-1 antagonist (10mg/kg) were significantly different from the vehicle control group (p < 0.05) see Figure 2. It shows that the inhibitory effect of baicalein in combination with PD-1 antagonists on tumor is significantly better than that of each control group, and the anti-tumor effect has a significant synergistic effect.

The above are only the preferred embodiments of the present invention. It should be pointed out that for those skilled in the art, some improvements and modifications can be made without departing from the principles of the present invention, and these improvements and modifications should also be regarded as It is the protection scope of the present invention.

Claims

The composition is characterized in that it consists of baicalein and a PD-1 antagonist.
The composition according to claim 1, wherein the mass ratio of the baicalein and the PD-1 antagonist is (25-150):(5-10).
The composition according to claim 1, wherein the mass ratio of the baicalein and the PD-1 antagonist is 25:5.
The application of the composition of any one of claims 1 to 3 in inhibiting the growth of tumor volume.
The use according to claim 4, wherein the inhibiting tumor volume growth comprises inhibiting the growth and/or proliferation of tumor cells.
Application of the composition of any one of claims 1 to 3 in the treatment of tumors.
Application of baicalein in enhancing the response rate of tumor patients to PD-1 antagonists.
Application of baicalein and PD-1 antagonists in the treatment of tumors.
The use according to claim 6 or 8, wherein the tumor treatment comprises inhibiting tumor volume growth and/or inhibiting tumor cell migration.
The application according to any one of claims 4 to 9, wherein baicalein plays a role in treating tumors by enhancing the response rate of tumor patients to PD-1 antagonists.
The use according to any one of claims 4-10, wherein the effective dose of baicalein is 25-150 mg/kg, and the effective dose of the PD-1 antagonist is 5-10 mg/kg.
A medicament for treating tumors, characterized by comprising the composition of any one of claims 1-3.
The medicine according to claim 12, characterized in that, in the medicine, baicalein and PD-1 antagonist are respectively made into preparations with pharmaceutically acceptable auxiliary materials, and the dosage forms of the preparations are tablets, capsules, Pills, granules, decoctions, ointments, lotions, oral liquids, injections, drop pills or syrups.
The composition according to any one of claims 1 to 3 or the application of claim 4 to 11 or the medicament according to any one of claims 12 to 13, wherein the PD-1 antagonist is PD-1 Monoclonal antibodies or antigen-binding fragments thereof.
The composition according to any one of claims 1 to 3 or the application of claim 4 to 11 or the medicine according to any one of claims 12 to 13, wherein the tumor is gastric cancer, intestinal cancer, and melanoma , breast cancer, esophageal cancer, lung cancer, ovarian cancer, cervical cancer, kidney cancer, bladder cancer, pancreatic cancer, prostate cancer, non-Hodgkin lymphoma, chronic lymphoma leukemia, multiple myeloma, acute myeloid leukemia or acute Lymphoma leukemia.