CN115837042A - Application of angelica sinensis blood-enriching decoction in preparation of medicine for enhancing curative effect of immune checkpoint inhibitor - Google Patents
Application of angelica sinensis blood-enriching decoction in preparation of medicine for enhancing curative effect of immune checkpoint inhibitor Download PDFInfo
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Abstract
The invention discloses application of angelica sinensis blood-enriching decoction in preparation of a medicine for enhancing curative effect of an immune checkpoint inhibitor, wherein in an animal model of tumors, the angelica sinensis blood-enriching decoction can promote CD8 in tumor tissues or draining lymph nodes + The Chinese angelica blood-enriching soup can obviously enhance the anti-late-stage tumor curative effect of the immune checkpoint inhibitor, shows synergistic effect by combining the Chinese angelica blood-enriching soup and the immune checkpoint inhibitor, and can achieve good treatment effect on the late-stage tumor, so the Chinese angelica blood-enriching soup can be applied to the preparation of the medicine for enhancing the curative effect of the immune checkpoint inhibitor. The invention provides a new thought and theoretical basis for the research of treating advanced tumors. The angelica sinensis blood-enriching soup is a natural traditional Chinese medicine component, has no toxic or side effect, wide raw material sources and low price, reduces the treatment cost of tumors, and solves the problem that patients with advanced tumors are easy to have immunityThe check point inhibitor has good application prospect due to the drug resistance problem.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to application of angelica sinensis blood-enriching decoction in preparation of a medicine for enhancing curative effect of an immune checkpoint inhibitor.
Background
Malignant tumor is one of the main causes of death of the global population, and seriously harms human life health. In recent years, immunotherapy has achieved remarkable clinical effects, and new eosin is brought to the field of tumor therapy. In particular, the clinical use of numerous immunotherapeutic drugs, typified by PD1/PDL1 immune checkpoint inhibitors, has extended the survival of numerous tumor species due to their broad antitumor activity. Immune Checkpoint Inhibitors (ICIs) therapy reactivates T cells by blocking immunosuppressive pathways (such as PD-L1/PD-1 and CTLA-4/B7-1) hijacked by tumor cells, reactivates the recognition and removal capacity of the immune system of an organism to the tumor cells, has the advantages of good curative effect and lasting response, and initiates a new era of tumor therapy. Immune checkpoint inhibitors have become the standard treatment for a variety of malignancies, including malignant melanoma, renal cell carcinoma, and the like. Not all patients can benefit from the treatment, the overall objective remission rate of patients receiving the single-dose treatment of the immune checkpoint inhibitor is only about 20%, and even in some patients with good initial immune treatment response, relapse can be caused by acquired resistance, and the poor curative effect of the immune checkpoint inhibitor on patients with advanced tumors is a clinical problem to be solved at present. Clinical data also indicate that tumor patients with anemia (tumor-associated anemia) that is complicated prior to treatment have difficulty in benefiting from PD1/PD-L1 antibody treatment, often presenting treatment resistance; the pre-treatment hemoglobin level is an independent prediction index of the treatment efficacy of the PD1/PD-L1 antibody of a tumor patient, and the resistance or drug resistance of the patient to an immune checkpoint drug is a clinical bottleneck problem which needs to be solved at present. Therefore, how to enhance the therapeutic effect of immune checkpoint inhibitors is critical.
In order to solve the above problems, there is a great need to develop a combination strategy that potentiates the efficacy of immune checkpoint inhibitors. The current mainstream combination strategies comprise treatment means such as surgery, chemotherapy, radiotherapy and targeted therapy and the like and the combination therapy of the ICIs, can generate synergistic effect, enhance the durability and curative effect of the ICIs and overcome the problem of low response rate of the ICIs. For example, the patent CN106963948A discloses the combined use of apatinib and a PD-1 inhibitor, and animal experiment data recorded in the specification proves that the combined use of apatinib and PD-1 inhibitor has a remarkable effect on the treatment of colon cancer. The invention patent CN105960415A discloses the combined use of axitinib and a PD-1 inhibitor, and the clinical test scheme described in the specification shows that the combined use of the axitinib and the PD-1 inhibitor is expected to have a remarkable treatment effect on renal cell carcinoma. Although these combination therapies may improve the efficacy and response rate of ici to some extent, toxicity is often increased, toxic side effects are greater, the incidence of adverse reactions is increased, and the physiological function and quality of life of the patient are reduced.
Immunological examinationSpot inhibitors promote immune cells (particularly CD 8) + T cells) killing the tumor, thereby improving the exhaustion state of T cells, is the key to its therapeutic effect. However, if the number of immune cells surrounding a tumor cell is lacking or the immune cell function of recognizing the tumor cell is disabled, even if the function that the immune checkpoint inhibitor can improve is exhausted, the therapeutic resistance or drug resistance of the body thereto cannot be avoided. While the inability of antigen presenting cells and the lack of T cells around tumors are important reasons for the development of resistance or tolerance of immune checkpoint drugs in patients with advanced tumors. Thus enhancing the number of T cells surrounding a tumor or facilitating the presentation of tumor antigens by antigen presenting cells is an important breakthrough in improving the resistance or tolerance of immune checkpoint inhibitor therapy.
Disclosure of Invention
Aiming at the defects of the prior art, the technical problems to be solved by the invention are as follows: how to provide the application of the angelica sinensis blood-enriching decoction in preparing the medicine for enhancing the curative effect of the immune checkpoint inhibitor, greatly improve the curative effect of the immune checkpoint inhibitor as an anti-tumor medicine, provide a new treatment scheme and a new thought for the treatment of late-stage tumors, and solve the problems that the curative effect of the immune checkpoint inhibitor on patients with the late-stage tumors is poor and the like.
In order to solve the technical problems, the invention adopts the following technical scheme: an application of a Chinese angelica blood-enriching decoction in preparing a medicament for enhancing the curative effect of an immune checkpoint inhibitor is disclosed, wherein the Chinese angelica blood-enriching decoction comprises astragalus and Chinese angelica.
Preferably, the mass ratio of the astragalus to the angelica is 1-10: 1.
preferably, the immune checkpoint inhibitor comprises one or more of a PD-L1 antibody, a PD-1 antibody, PD-1 and PD-L1.
The invention also aims to provide a pharmaceutical composition for resisting the advanced tumor, which comprises the angelica sinensis blood-enriching decoction and an immune checkpoint inhibitor.
The invention also aims to provide application of the pharmaceutical composition in preparing a medicament for treating advanced tumors.
Preferably, the tumor is one or more of colon cancer, melanoma, gastric cancer, esophageal cancer, lung cancer, breast cancer, ovarian cancer, liver cancer, pancreatic cancer, cervical cancer and prostate cancer.
Preferably, the angelica sinensis blood-enriching decoction and the immune checkpoint inhibitor are administered to a tested patient, and the angelica sinensis blood-enriching decoction comprises the following components in a mass ratio of 1-10: 1 radix astragali and radix Angelicae sinensis.
Preferably, the angelica sinensis blood-enriching decoction and the immune checkpoint inhibitor can be simultaneously, separately or sequentially administered.
Compared with the prior art, the invention has the following beneficial effects:
1. the invention discovers for the first time that the angelica sinensis blood-enriching decoction can enhance the curative effect of the immune checkpoint inhibitor on the advanced tumor, and in an animal model of the tumor, the angelica sinensis blood-enriching decoction can promote CD8 in tumor tissues or drainage lymph nodes + The Chinese angelica blood-enriching decoction can obviously enhance the anti-late-stage tumor curative effect of the immune checkpoint inhibitor, shows synergistic effect by combining the Chinese angelica blood-enriching decoction and the immune checkpoint inhibitor, can achieve good tumor treatment effect, and solves the problem that the immune checkpoint inhibitor has poor curative effect on patients with late-stage tumors. Therefore, the angelica sinensis blood-enriching decoction combined with the immune checkpoint inhibitor can be applied to preparing the medicine for resisting the advanced tumor. The invention provides a new thought and theoretical basis for the research of treating advanced tumors, and has great significance.
2. The angelica sinensis blood-enriching soup is a natural traditional Chinese medicine component, has higher safety and smaller toxic and side effects, can ensure better physiological function and life quality for patients, has wide raw material sources and low price, reduces the treatment cost of tumors, and has good application prospect.
Drawings
FIG. 1 is a schematic representation of the treatment process of mice of different experimental groups in the middle and late stage tumor experiment of example 1.
Fig. 2 is a photograph of tumor volume over time for different treatment groups of mice in an advanced tumor experiment.
FIG. 3 is a graph of tumor volume versus time for different treatment groups of mice in an advanced tumor experiment.
FIG. 4 is a graph of tumor size over time for different treatment groups of mice in an early stage tumor experiment; a is a schematic representation of the treatment process, B is the tumor volume, C is a photograph of the tumor size, and D is the tumor weight.
FIG. 5 is a flow cytogram of spleens of mice of different treatment groups in example 2; a is CD8 + Proportion of T cells, B is CD4 + Proportion of T cells.
FIG. 6 is a flow cytogram of spleens of mice of different treatment groups in example 2; a is CD8 + The ratio of the activating factor IFN-gamma + in T cells, B is CD8 + The proportion of the activating factor TNF-. Alpha. + in T cells.
FIG. 7 is a graph showing CD8 in peripheral blood of mice of different treatment groups in example 3 + Proportion of T cells.
FIG. 8 is a graph showing the ratio of lymphocytes (A) in tumor cells and CD8 in mice of different treatment groups in example 3 + Proportion of T cells (B) and activated CD8 + Proportion of T cells (C).
FIG. 9 is CD8 in tumor tissue draining lymph nodes of mice of different treatment groups in example 4 + Proportion of T cells (A) and activated CD8 + Proportion of T cells (B).
FIG. 10 is a schematic view showing the treatment process of mice of different experimental groups in example 5.
FIG. 11 shows the ratio of OT-1T cells to CD8+ T cells (A), the number of OT-1T cells (B), and the ratio of activated OT-1T cells (C/D) in the tumor tissue draining lymph nodes of mice of different treatment groups in example 5.
Detailed Description
The present invention will be described in further detail with reference to examples.
MC38 cells (colon cancer, 5X 10) were used in the following examples 5 ) Respectively inoculating the seeds on the backs of 6-week-old C57BL/6 mice to establish a subcutaneous tumor-bearing model of the mice.
Preparing the angelica blood-enriching soup: 20g of angelica and 100g of astragalus. Decocting in water, and concentrating to 1.5g crude drug/ml for use (according to pharmacopeia, astragaloside IV and ferulic acid are used as quality control indexes, and content of astragaloside IV and ferulic acid is detected by LC-MS).
Example 1
In the experiment of intervening late stage tumor, tumor-bearing mice are divided into a control group (CON), a Chinese angelica blood-enriching Decoction Group (DGBX) and an anti-PD-L1 group, and the Chinese angelica blood-enriching Decoction (DGBX) is combined with the anti-PD-L1 group. Angelica sinensis blood enriching Shang Zu from the 15 th day of tumor bearing, the angelica sinensis blood enriching decoction (30 g crude drug/kg body weight) is administrated with 1 times per day by intragastric administration and subcutaneous injection of PBS; the anti-PD-L1 group was gavaged with physiological saline 1 time per day from the 15 th day of tumor bearing and was injected subcutaneously with PD-L1 antibody (200. Mu.g/200. Mu.l/mouse, once every 2 days, 4 times in total) from the 15 th day of tumor bearing; the angelica sinensis blood-enriching decoction and anti-PD-L1 group is administrated with 1 times of gastric lavage of angelica sinensis blood-enriching decoction (30 g crude drug/kg body weight) every day from the 15 th day of tumor bearing, and is injected with PD-L1 antibody (200 mug/one, once every 2 days for 4 times) subcutaneously from the 15 th day of tumor bearing, and the control group mice are administrated with the same dosage and frequency of physiological saline lavage or PBS subcutaneous injection (as shown in figure 1). Tumor volumes of tumor-bearing mice in each group were measured every 2 days, and the mice in each group were sacrificed on day 27, and the results are shown in fig. 2 and 3.
In the intervention experiment of early tumors, tumor-bearing mice were divided into a Control group (Control), a angelica sinensis blood-enriching Decoction Group (DGBX) and an anti-PD-L1 group, the angelica sinensis blood-enriching Shang Zu was administered with an angelica sinensis blood-enriching decoction (30 g crude drug/kg body weight) 1 time per day from the tumor-bearing day 0 and PBS was subcutaneously injected every 2 days from the day 7 (4 times total injection), the anti-PD-L1 group was subcutaneously injected with PD-L1 antibody (200 μ g/200 μ L/mouse, once every 2 days, 4 times total injection) from the tumor-bearing day 7 and physiological saline was administered from the day 0 (as in fig. 4A), the Control group mice were administered with an equal dose of physiological saline on the day 0 and PBS was subcutaneously injected with an equal frequency and dose on the day 7, the tumor-bearing mice were measured every 2 days, the tumor-bearing mice of each group were sacrificed on the day 25, and the weights of the tumors were weighed, and the results are shown in fig. 4.
As can be seen from FIGS. 2 and 3, in the experiment of intervention of advanced tumor, the volume of the tumor in mice of the Dang Gui Buxue Tang group was not significantly different from that of the tumor in mice of the control group, which indicates that the tumor control in the advanced tumor-bearing stage is not effective by using Dang Gui Buxue Tang aloneTumor growth; in the early intervention experiment of tumor, the Chinese angelica decoction for enriching blood can effectively control the growth of tumor by singly using, and the curative effect of the Chinese angelica decoction for enriching blood is equivalent to that of the immune checkpoint inhibitor (as shown in figure 4B, figure 4C and figure 4D); this may be early tumor CD8 + The function of T is not seriously exhausted, and the advanced tumor CD8 is obtained + The function exhaustion of T cells is already serious, so that the angelica sinensis blood-enriching decoction alone has curative effect on tumors in the early stage of the tumors, but has no curative effect in the late stage of the tumors. Surprisingly, however, in the experiment of tumor advanced intervention, the angelica sinensis hematinic decoction combined with anti-PD-L1 acts on tumor-bearing mice, and all the mice in the group show smaller tumor volumes which are respectively reduced by 75% and 50% compared with the tumor volumes of a control group and an anti-PD-L1 group (as shown in figure 3), and have significant difference. This indicates that, although the Dang Gui Xue Tang can not exert the anti-tumor effect in the advanced stage of tumor, dang Gui Xue Tang can significantly enhance the anti-tumor effect of PD-L1 antibody.
Example 2
In the experiment of the intervention of the advanced Tumor, tumor-bearing mice are divided into a Control group (Control) and a Chinese angelica blood-enriching Decoction Group (DGBX), and normal Tumor-free mice are used as a negative Control (Tumor-free). On the 15 th day of the tumor, the mice in the angelica sinensis blood-enriching decoction group are filled with the angelica sinensis blood-enriching decoction (30 g crude drug/kg body weight) for intragastric administration 1 time per day, and the control group and the negative control group are filled with normal saline for intragastric administration. Mice of each group were sacrificed on day 27 of tumor bearing, and spleens of mice were removed for CD4 + T and CD8 + T cells were detected by flow cytometry, and statistical significance of the differences was determined using independent sample T-tests (mean ± s.d.. P.)<0.01,***P<0.001 Results are shown in fig. 5 and 6.
As can be seen from FIG. 5, CD4 in spleen of control group mice was compared with that of negative control group + And CD8 + The proportion of T cells is obviously reduced, and the CD4 in the spleen of the mouse intervened by the angelica sinensis blood-enriching decoction + And CD8 + The proportion of T cells is obviously improved. As can be seen from FIG. 6, CD8 in spleen of control group mice was compared with that of negative control group + The function of T cells is obviously reduced, and the CD8 in the spleen of the mouse intervened by the angelica sinensis blood-enriching decoction + The functionality of T cells is recoveredAnd the level of the compound can reach the level of a normal mouse. It is demonstrated that the Chinese angelica decoction for enriching blood can up-regulate the CD4 of spleen of tumor organism + And CD8 + Number and function of T cells.
Example 3
In the experiment of intervening in the late stage tumor, tumor-bearing mice were divided into a Control group (Control) and a angelica sinensis blood-enriching Decoction Group (DGBX). On day 15 of the tumor, the mice in the angelica sinensis blood-enriching decoction group are filled with the angelica sinensis blood-enriching decoction (30 g crude drug/kg body weight) for intragastric administration 1 time per day, and the control group is filled with normal saline for intragastric administration. Mice from each group were sacrificed on day 27 and peripheral blood and tumor tissue were removed for flow cytometry and statistical significance of the differences was determined using independent sample t-tests (mean ± s.d. P <0.05, P < 0.01) with the results shown in fig. 7 and 8.
As can be seen from the figure, compared with the control group, the mice intervened by the angelica sinensis blood-enriching decoction are not only CD8 in peripheral blood + The ratio of T cells in (A), or CD8 in tumor tissue + The proportion of T cells is obviously increased, and the proportion of T lymphocytes in tumor cells of mice intervened by the angelica sinensis blood-enriching decoction and the activated CD8 + The number of T cells in (a) also increases significantly. It is demonstrated that the Dang Gui Buxue Tang can up-regulate the peripheral blood of tumor and the CD8 in tumor tissue + Number and function of T cells.
Example 4
In the experiment of intervening in the late stage tumor, tumor-bearing mice were divided into a Control group (Control) and a angelica sinensis blood-enriching Decoction Group (DGBX). On the 15 th day of the tumor, the mice in the angelica blood-enriching decoction group are filled with the angelica blood-enriching decoction (30 g crude drug/kg body weight) for intragastric administration for 1 time every day, and the mice in the control group are filled with normal saline for intragastric administration. Groups of mice were sacrificed on day 27 with tumor, and the paratumoral draining lymph nodes were removed for detection by flow cytometry. Statistical significance of the differences was determined using t-test on independent samples (mean ± s.d.. P <0.01,. P < 0.001) and the results are shown in fig. 9.
As can be seen from the figure, compared with the control group, the CD8 in the mouse draining lymph node intervened by the angelica sinensis blood-enriching decoction + Has significantly increased T cell ratio and activates CD8 + The number of T cells in (a) also increases significantly. It is demonstrated that the blood-enriching decoction of Chinese angelica can up-regulate the draining lymph node of tumor organismMiddle CD8 + Number and function of T cells.
Example 5
The MC38 subcutaneous tumor-bearing mice were divided into a Control group (Control) and a Danggui Buxue Decoction Group (DGBX). On day 7 of tumor, the mice in the group of DANGGUIBUXUE decoction are filled with DANGGUIBUXUE decoction (30 g crude drug/kg body weight) for intragastric administration 1 time per day, and the control group is filled with normal saline for intragastric administration. Gavage was completed on day 13 of tumor bearing, but two groups of mice were given terminal intravenous injections of OT-1T cells, and each group of mice was sacrificed on day 17 of tumor bearing (as shown in fig. 10), and the paratumoral draining lymph nodes were removed and examined by flow cytometry. Statistical significance of the differences was determined using t-test on independent samples (mean ± s.d.. P <0.01,. P < 0.001) and the results are shown in fig. 11.
As can be seen from the figure, OT-1T cells in the draining lymph nodes of the mice intervened by the Angelica sinensis decoction for enriching blood account for CD8, compared with the control group + The proportion of T cells is obviously increased, the number of OT-1T cells is obviously increased, and the proportion of activated OT-1T cells is obviously increased. It is demonstrated that Dang Gui Bu Tang can enhance the antigen presentation process.
In conclusion, in an animal model of tumor, the angelica sinensis blood-enriching decoction can obviously enhance the anti-late-stage tumor curative effect of the immune checkpoint inhibitor. The reason is that the mechanism of the angelica sinensis blood-enriching decoction which exerts the sensitization curative effect is to promote CD8 in tumor tissues or drainage lymph nodes + Proliferation and activation of T cells and promotion of antigen presentation of tumor cells by the body. Therefore, the angelica sinensis blood-enriching decoction can solve the problem that the immune checkpoint inhibitor has poor curative effect on patients with advanced tumors.
The above description is only exemplary of the present invention and should not be taken as limiting, and any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. An application of a Chinese angelica blood-enriching decoction in preparing a medicament for enhancing the curative effect of an immune checkpoint inhibitor is disclosed, wherein the Chinese angelica blood-enriching decoction comprises astragalus and Chinese angelica.
2. The application of claim 1, wherein the mass ratio of the astragalus to the angelica is 1-10: 1.
3. the use of claim 1, wherein the immune checkpoint inhibitor comprises one or more of a PD-L1 antibody, a PD-1 antibody, PD-1, and PD-L1.
4. A pharmaceutical composition for treating advanced tumors, comprising the Angelica sinensis blood-enriching decoction of any one of claims 1 to 3 and an immune checkpoint inhibitor.
5. Use of the pharmaceutical composition of claim 4 for the preparation of a medicament for the treatment of advanced tumors.
6. The use of claim 5, wherein the tumor is one or more of colon cancer, melanoma, gastric cancer, esophageal cancer, lung cancer, breast cancer, ovarian cancer, liver cancer, pancreatic cancer, cervical cancer, and prostate cancer.
7. The use of claim 5, wherein said Angelica sinensis decoction for tonifying blood and said immune checkpoint inhibitor are administered to said subject, said Angelica sinensis decoction for tonifying blood comprises a mass ratio of 1-10: 1 radix astragali and radix Angelicae sinensis.
8. The use of claim 7, wherein the angelicae sinensis blood-enriching decoction and the immune checkpoint inhibitor are administered simultaneously, separately or sequentially.
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