CN112274525B

CN112274525B - Chemotherapy pharmaceutical composition and application thereof

Info

Publication number: CN112274525B
Application number: CN202011411820.1A
Authority: CN
Inventors: 高宁; 姜秀星; 龚其海; 李志强; 申立文; 王梅
Original assignee: Zunyi Medical University
Current assignee: Zunyi Medical University
Priority date: 2020-12-04
Filing date: 2020-12-04
Publication date: 2022-04-08
Anticipated expiration: 2040-12-04
Also published as: CN112274525A

Abstract

The invention relates to the technical field of biological medicines, in particular to a chemotherapeutic medicine composition and application thereof. A chemotherapeutic composition comprises cepharanthine and epirubicin. Before administration of epirubicin, cepharanthine is administered to synergistically enhance the cancer cell inhibitory effect of epirubicin. The scheme solves the technical problems that the epirubicin has limited curative effect and the cancer patients are easy to generate adverse reaction and drug resistance after long-term use. The composition can be applied to clinical practice of treatment of various cancers, reduces adverse reactions of epirubicin and simultaneously prevents the appearance of drug resistance of epirubicin.

Description

Chemotherapy pharmaceutical composition and application thereof

Technical Field

The invention relates to the technical field of biological medicines, in particular to a chemotherapeutic medicine composition and application thereof.

Background

Chemotherapy is one of the methods for treating malignant tumors (cancers), and is a treatment method that uses chemical drugs to prevent the proliferation, infiltration and metastasis of cancer cells until the cancer cells are finally killed. Epirubicin (Epirubicin), also known as Epirubicin, is more potent than Doxorubicin (Doxorubicin), but less toxic to the heart, and is one of the currently clinically used chemotherapeutic agents. The epirubicin injection is clinically used for treating breast cancer, ovarian cancer, digestive tract cancer (such as advanced gastric cancer and advanced colorectal cancer), malignant lymphoma, lung cancer, malignant melanoma and the like. Just, epirubicin, like other chemotherapeutic drugs, is not very selective, and inevitably damages normal cells of the human body while killing cancer cells, thereby causing adverse reactions of the drugs. The time for treating malignant tumor is relatively long, and during the long-term treatment, the drug resistance or drug resistance is generated gradually. How to improve the curative effect of epirubicin without increasing the dosage of epirubicin, thereby reducing the adverse reaction of epirubicin and simultaneously preventing the occurrence of drug resistance is a difficult point to be overcome when using therapeutic drugs such as epirubicin and the like to treat cancers.

Disclosure of Invention

The invention aims to provide a chemotherapeutic medicine composition to solve the technical problems that the curative effect of epirubicin is limited, and the long-term use of epirubicin by cancer patients is easy to cause adverse reaction and drug resistance.

In order to achieve the purpose, the invention adopts the following technical scheme:

a chemotherapeutic composition comprises cepharanthine and epirubicin.

The principle and the advantages of the scheme are as follows: epirubicin (cas: 56363-09-1) is a commonly used anticancer chemotherapeutic drug in clinic, and cepharanthine is administered before Epirubicin administration, so that the cancer cell inhibition effect of Epirubicin can be synergistically enhanced. Cepharanthine (Cepharanthine, CEP, cas: 481-49-2) is an isoquinoline alkaloid extracted from the root of Stephania japonica Thunb of Menispermaceae, and is mainly used for increasing leukocyte in clinic with little toxic and side effects. Under the condition of normal dosage, the cepharanthine is a medicine basically without toxic and side effects, only occasional gastrointestinal reactions are seen in clinic, and no obvious toxic and side effects are seen. The use of cepharanthine can reduce the dosage of epirubicin on the basis of ensuring the drug effect of epirubicin, reduce adverse reaction caused by excessive use of epirubicin or avoid the generation of drug resistance of epirubicin in human body.

In conclusion, cepharanthine has the effect of being capable of cooperating with the clinical chemotherapy drug epirubicin to improve the effect of inhibiting and killing various tumor cells. Experiments show that for breast cancer MDA-MB-231 cells, MCF-7 cells, BT549 cells, leukemia K562 and U937 cells, stomach cancer SGC-7901 cells, lung cancer A549 cells and esophageal cancer Eca109 cells, under the condition of using the cepharanthine, the effects of inhibiting and killing the tumor cells by the epirubicin are greatly improved, and the effect of the cepharanthine in combination with the epirubicin is obviously better than the effect of the cepharanthine in combination with the doxorubicin and the effect of the epirubicin in combination with the chloroquine.

Further, the mass ratio of the cepharanthine to the epirubicin is 0.97: 0.032.

By adopting the technical scheme, the cepharanthine and epirubicin in the mass ratio can form effective inhibition effect on tumor cells. When the combination of the cepharanthine and the epirubicin is used for treating tumors, the mass ratio of the total dose of the cepharanthine to the total dose of the epirubicin is 0.97: 0.032.

Further, the single dose of cepharanthine in human is 0.97mg/kg · bw.

Further, the human single dose of epirubicin is 0.16 mg/kg-bw.

By adopting the technical scheme, the single human body dose of the cepharanthine and the epirubicin is obtained through animal experiment results and conversion. In Experimental example 2, the single dose of cepharanthine in nude mice was 12mg/kg bw, and the single dose of epirubicin in nude mice was 2mg/kg bw. The conversion is based on the conversion ratio (1: 12.3) of the injected Dose of human to mouse (i.e., experimental-verified nude mice) required to achieve the same biological effect, as given by the U.S. Food and Drug Administration (FDA) in the estimation of Safe Dose for Healthy Adult subjects in Clinical Trials of drugs (estimation of Safe Dose for Therapeutics in human subjects).

Further, the ratio of the administration frequency of cepharanthine and epirubicin in one treatment course was 5: 1.

By adopting the technical scheme, the use of the low-toxicity cepharanthine can reduce the administration frequency of epirubicin and reduce the harm of chemotherapeutic drugs to human bodies.

Further, the routes of administration of cepharanthine and epirubicin are both intravenous administration.

By adopting the technical scheme, intravenous injection administration is a conventional administration mode of the cepharanthine and epirubicin. In the case of injection of the epirubicin injection solution in the combination drug of the present invention, the administration is actually discontinuous in the same manner as that of the epirubicin injection solution alone. Because the epirubicin drug has longer metabolism time in human body, the non-continuous administration mode can reduce toxic and side effects. Even though the dosage of epirubicin can be reduced after the drug combination is formed by using the synergy of cepharanthine, the administration mode is still adopted, which accords with the traditional administration habit and is more convenient. When the medicine combination is applied, the step of injecting the cepharanthine injection is added, and no other special operation method is available. The actual dosage of the cepharanthine in the pharmaceutical composition of the invention is much more than that of epirubicin. The combination of experimental verification shows that the combination of cepharanthine and epirubicin can inhibit and kill various malignant tumor cells, reduce the dosage, shorten the treatment period and reduce the occurrence of toxic and side effects. Even in the contrary, under the medium action of the epirubicin serving as the conventional clinical chemotherapy medicament, the cepharanthine which is found by the inventor of the application and has the effect of effectively blocking autophagy degradation of various tumor cells also has the effect of killing various malignant tumor cells with reliable effect and practical value.

Further, the application of a chemotherapeutic drug composition as an anti-cancer drug is disclosed, wherein the chemotherapeutic drug composition comprises cepharanthine and epirubicin.

By adopting the technical scheme, the cepharanthine and epirubicin have the effect of synergistically enhancing the killing effect of cancer cells, and can be applied to the preparation of anti-cancer drugs.

Further, the anticancer drug is used for treating triple negative breast cancer.

By adopting the technical scheme, the combination of the cepharanthine and the epirubicin has obvious inhibition effect on human triple negative breast cancer cells MDA-MB-231 and BT549 cells, and has weaker sensitization effect on non-triple negative breast cancer cells MCF-7 cells, so that the cepharanthine and the epirubicin can be used for treating triple negative breast cancer. The composition of cepharanthine and epirubicin shows good triple negative breast cancer cell inhibition effect. The triple negative breast cancer refers to the breast cancer with negative cancer tissue immunohistochemical examination results of Estrogen Receptor (ER), progestational hormone receptor (PR) and proto-oncogene Her-2, and has poor prognosis and high death risk.

Further, in one treatment course, the administration frequency of cepharanthine is 20 times, and the administration frequency of epirubicin is 4 times.

By adopting the technical scheme, the use of the low-toxicity cepharanthine can reduce the administration times of epirubicin.

Further, within a course of treatment, the frequency of administration of cepharanthine is once a day and the frequency of administration of epirubicin is once every 5 days.

By adopting the technical scheme, the use of the low-toxicity cepharanthine can reduce the administration frequency of epirubicin.

Drawings

FIG. 1 is a graph showing the inhibition curves of epirubicin and epirubicin in combination with cepharanthine in Experimental example 1 (the subject was breast cancer cells, and the concentration of cepharanthine was constant).

FIG. 2 is a graph showing the inhibition curves of cancer cells by using cepharanthine alone and cepharanthine in combination with epirubicin in Experimental example 1 (the subject is breast cancer cells, and the concentration of epirubicin is constant).

FIG. 3 is a statistical chart of the cancer cell inhibitory effects of epirubicin and doxorubicin, respectively, in combination with cepharanthine in Experimental example 1.

FIG. 4 is a graph showing the statistics of the inhibition effect of the cancer cells by the combination of cepharanthine and chloroquine, respectively, with epirubicin in Experimental example 1.

FIG. 5 is a statistical graph showing the cancer cell inhibitory effect of epirubicin in combination with cepharanthine of Experimental example 1 (against cells other than breast cancer cells).

Fig. 6 is a photograph of mice of different experimental groups of experimental example 2 (showing tumor volume).

FIG. 7 is a graph showing the change in tumor volume of experimental mice in Experimental example 2.

FIG. 8 is a graph showing the change in body weight of experimental mice in Experimental example 2.

Detailed Description

The following is further detailed by way of specific embodiments:

example 1:

the chemotherapeutic medicine composition consists of cepharanthine and epirubicin, and more specifically, the medicine composition comprises cepharanthine and chemotherapeutic epirubicin. The structural formula of cepharanthine is shown in formula (I), and the structural formula of epirubicin is shown in formula (II).

After understanding that the pharmaceutical composition provided by the present invention can improve the therapeutic effect and reduce adverse reactions, when the pharmaceutical composition of the present invention is used to avoid drug resistance or drug resistance, it is fully within the ability of those skilled in the art to determine the ratio of the two components cepharanthine and epirubicin for a specific malignant tumor by only limited routine tests. Aiming at several malignant tumor cells selected during experimental verification, the medicine composition is preferably prepared according to the proportion, and in one treatment course, the total medicine quantity of the cepharanthine and the total medicine quantity of the epirubicin are 0.97: 0.032. The above dose ratio is converted from the experimental animal dose (experimental example 2). The conversion is based on the conversion ratio (1: 12.3) of the injected Dose of human to mouse (i.e., experimental-verified nude mice) required to achieve the same biological effect, as given by the U.S. Food and Drug Administration (FDA) in the estimation of Safe Dose for Healthy Adult subjects in Clinical Trials of drugs (estimation of Safe Dose for Therapeutics in human subjects).

When the medicine combination of the scheme is applied to treatment, the cepharanthine and epirubicin are both administrated in a divided mode (but the ratio of the total medicine amount is kept to be 0.97: 0.032), and the specific application method is as follows:

the injection is prepared by injecting the cepharanthine injection once a day through a vein according to the dose of the cepharanthine and the weight of a patient which is 0.97mg/kg (normal saline or 5 percent glucose injection is needed to prepare the cepharanthine injection, the normal saline is used in the embodiment);

② on the day of cepharanthine injection according to the step (i), according to the dosage of epirubicin to patient's weight being 0.16mg/kg (using normal saline or 5% glucose injection to prepare epirubicin injection, the glucose injection used in this example), the epirubicin injection is injected once by vein;

thirdly, when the sixth day comes, according to the dosage of the second step, the epirubicin injection with the same dosage is injected again through veins; circulating according to the steps of the first step and the third step;

fourthly, from the twenty-first day, whether to circulate according to the methods of the first step to the third step, the circulation times and the intermediate medicine stopping time are determined according to the bearing capacity and the treatment effect of the body of the patient.

It is clear to those skilled in the art that the concentration of the injection should be determined according to the tolerance of the patient, the "number of cycles" in step (iv) may be generally 1-4 times, and the "intermediate stopping time" may be generally 1-2 weeks; for patients with particularly poor health, it is even possible to use only steps (i) and (ii) as a medication cycle or a course of treatment (i.e. to suspend medication from the sixth day onwards).

Experimental example 1: cell experiments

1. Cell selection and active culture

The experimental selection is that human breast cancer cells MDA-MB-231, MCF-7 and BT549, human leukemia cells K562 and U937, human gastric adenocarcinoma cells SGC-7901, human lung cancer cells A549 and human esophageal carcinoma cells Eca 109. Wherein, human breast cancer cells MDA-MB-231, MCF-7, BT549 cells, human gastric adenocarcinoma SGC-7901 cells and human lung cancer A549 cells are actively cultured in a DMEM + 10% FBS complete culture medium; human leukemia cells K562, U937 cells, human esophageal cancer cells Eca109 were cultured in RPMI1640+ 10% FBS complete medium. The above cells are all at 37 deg.C and 5% CO₂And culturing in a cell culture box with saturated humidity.

2. Experimental methods and results

After each tumor cell line was stable, the cells were plated in 96-well plates with 5000 cells per well containing 90 μ L of complete medium, and then the cells were treated with the drug. MTT assay was performed on the treated cells to determine the effect of the drug on cell activity: after 48 hours of dosing, 20. mu.L of 5mg/mL thiazole blue (MTT) solution in Phosphate Buffered Saline (PBS) was added to each well and incubation was continued for 4 hours. Sucking the upper layer culture medium (human leukemia K562 and U937 cells firstly centrifugate the suspended cells to the bottom of the wells by a plate centrifuge), adding 150 mu L of dimethyl sulfoxide (DMSO) into each well for dissolving, dissolving for 10 minutes on a vibrator, detecting the absorbance value at A495 by an enzyme labeling instrument, and calculating the cell survival rate according to the following formula:

percent cell survival ═ (treatment absorbance-blank absorbance) ÷ (control absorbance-blank absorbance) × 100%.

The specific experimental grouping was as follows:

human breast cancer cells MDA-MB-231, MCF-7 and BT549 cells are grouped and treated in the following modes:

(1) control group: complete medium solution 10. mu.L.

(2) Epirubicin group: epirubicin solutions were prepared in complete medium at concentrations of 0, 1, 2, 4, 6, 8 μ M (μmol/L) and treated at 10 μ L per well to give final concentrations of 0, 0.1, 0.2, 0.4, 0.6, 0.8 μ M. The results of the experiment are shown in FIG. 1.

(3) Cepharanthine group: the whole culture medium is used to prepare cepharanthine solution with concentration of 0, 10, 20, 40, 60, 80 μ M (μmol/L), and 10 μ L of cepharanthine solution is added into each well to make the final concentration of the cepharanthine solution be 0, 1, 2, 4, 6, 8 μ M. The results of the experiment are shown in FIG. 2.

(4) A combination of drugs: epirubicin and cepharanthine combination drug group

Epirubicin and cepharanthine are administered in combination using two experimental protocols:

the first scheme is as follows: and (3) preparing epirubicin solutions with the concentrations of 0, 1, 2, 4, 6 and 8 mu M and cepharanthine mixed liquor with the concentrations of 20 mu M by using a complete culture medium, and adding 10 mu L of the mixed liquor into each hole for treatment so that the final acting concentrations of epirubicin are 0, 0.1, 0.2, 0.4, 0.6 and 0.8 mu M respectively, and the final acting concentration of cepharanthine is 2 mu M. Experimental results referring to fig. 1, epirubicin itself has a certain concentration-dependent cancer cell inhibitory effect, and the cancer cell inhibitory effect of epirubicin at the same concentration is enhanced by the action of cepharanthine.

Scheme II: and preparing a cepharanthine solution with the concentration of 0, 10, 20, 40, 60 and 80 mu M and an epirubicin mixed liquid medicine with the concentration of 2 mu M by using a complete culture medium, and adding 10 mu L of the mixed liquid medicine into each hole for treatment so that the final acting concentrations of the cepharanthine are respectively 0, 1, 2, 4, 6 and 8 mu M and the final acting concentration of the epirubicin is 0.2 mu M. The experimental results are shown in fig. 2, cepharanthine itself does not have a significant cancer cell inhibitory effect, and as the amount of cepharanthine increases, cepharanthine does not exhibit a cancer cell inhibitory effect. However, after the addition of 0.2 μ M of epirubicin, the composition had a significant effect of inhibiting cancer cells, which was not produced only by epirubicin alone, but by a synergistic effect of epirubicin and cepharanthine. Referring to fig. 1 and 2, cepharanthine alone had no significant effect on cell viability. Compared with the epirubicin group used alone, the tumor cell survival rate of the cepharanthine and epirubicin group is lower than that of the epirubicin group used alone to different degrees, and the combined effect of the two is not additive, but 1+1 is greater than 2.

By combining the experimental results shown in the figure 1 and the figure 2, the combination of the cepharanthine and the epirubicin has obvious inhibition effect on human triple-negative breast cancer cells MDA-MB-231 and BT549 cells, but has weaker sensitization effect on non-triple-negative breast cancer cells MCF-7 cells, so that the cepharanthine and the epirubicin can be used for treating triple-negative breast cancer. The composition of cepharanthine and epirubicin shows good triple negative breast cancer cell inhibition effect.

(5) Doxorubicin and cepharanthine combined drug group

And (3) preparing a doxorubicin solution with the concentration of 2 mu M and a cepharanthine mixed liquid medicine with the concentration of 20 mu M by using a complete culture medium, and adding 10 mu L of the mixed liquid medicine into each hole for treatment so that the final acting concentrations are 0.2 mu M and 2 mu M respectively. The experimental results are shown in fig. 3, the final concentration of cepharanthine group is 2 μ M; the final concentration of epirubicin group was 0.2 μ M; in the epirubicin + cepharanthine group, the final concentration of epirubicin is 0.2 μ M, and the final concentration of cepharanthine is 2 μ M; the final concentration of doxorubicin group was 0.2 μ M; in the doxorubicin + cepharanthine group, the final concentration of doxorubicin was 0.2 μ M and the final concentration of cepharanthine was 2 μ M. Doxorubicin and epirubicin are isomers, and the mechanism of action is direct intercalation between DNA nucleobase pairs, interfering with the transcriptional process, preventing mRNA formation, and thereby inhibiting DNA and RNA synthesis. The inventor researches and discovers that the inhibition effect of doxorubicin and epirubicin on cancer cells is equivalent when the doxorubicin and epirubicin are used singly, but the combined effect of the cepharanthine and epirubicin is obviously better than the effect of doxorubicin combined with the cepharanthine.

(6) Epirubicin and chloroquine combined drug group

And (3) preparing an epirubicin solution with the concentration of 2 mu M and chloroquine mixed liquor with the concentration of 40 and 200 mu M by using a complete culture medium, adding 10 mu L of the mixed liquor into each hole for treatment, and respectively enabling the epirubicin to have the final action concentration of 0.2 mu M, the chloroquine to have the final action concentrations of 4 and 20 mu M and the cepharanthine to have the final action concentration of 2 mu M in single application and combined application. Referring to fig. 4, 4 μ M chloroquine and 0.2 μ M epirubicin, when used in combination, did not significantly inhibit the growth of cancer cells; the combination of cepharanthine of 2 μ M and epirubicin of 0.2 μ M can obviously inhibit the growth of cancer cells; increasing the amount of chloroquine to 20 μ M, the synergistic effect of chloroquine and epirubicin in inhibiting cancer cells was still not as good as the combination of cepharanthine and epirubicin. Cepharanthine and Chloroquine (CQ) are both autophagy inhibitors, and chloroquine is currently the only clinically used autophagy inhibitor approved by the FDA. Although both have an autophagy-inhibiting effect, they have a very significant difference in the promoting effect on the cancer-inhibiting effect of epirubicin when both are used in combination with epirubicin, respectively.

Human gastric adenocarcinoma SGC-7901 cells, human lung cancer A549 cells, human leukemia K562 cells, U937 cells, human esophageal carcinoma Eca109 cells were grouped and treated as follows:

(1) control group: treating with 10 μ L of complete culture medium solution;

(2) epirubicin group: preparing epirubicin solution with concentration of 2 μ M (μmol/L) with complete culture medium, and treating each well with 10 μ L to make final concentration of 0.2 μ M;

(3) cepharanthine group: preparing 20 mu M (mu mol/L) cepharanthine solution with complete culture medium, and treating each well with 10 mu L; the final concentration of the drug is 2 mu M;

(4) epirubicin and cepharanthine combination drug group: and (3) preparing epirubicin solution with the concentration of 2 mu M and cepharanthine mixed liquor with the concentration of 20 mu M by using a complete culture medium, and adding 10 mu L of the mixed liquor into each hole for treatment so that the final acting concentrations are 0.2 mu M and 2 mu M respectively.

The experimental result is shown in fig. 5, the combination of epirubicin and cepharanthine can obviously inhibit the activity of human gastric adenocarcinoma SGC-7901 cells, human lung cancer A549 cells, human leukemia K562 cells, U937 cells and human esophageal carcinoma Eca109 cells.

In addition, in fig. 3 to 5, the ordinate represents the cell survival rate, i.e., the cell survival rate of the treated group relative to the control group. The abscissa represents various human tumor cell lines (note: represented by the distinguishing symbols of the corresponding tumor cells only). The values corresponding to the bars are the average values of the groups, and the upper and lower deviations are indicated by the "I" symbols in the figures.

For the experimental results of fig. 1 to 5, in order to prove that the verification conclusion is accurate and reliable, statistical calculation and analysis are performed on the sample during verification. In the figure ". mark" means p < 0.01, where "p" is the boundary symbol of a statistically acceptable error, with a value of p ≦ 0.05. Under the condition that p is calculated to be less than 0.01, the survival rate of the tumor cells of the cepharanthine and epirubicin group is determined to be less than that of the tumor cells of the epirubicin alone, and the possibility of error occurrence is low, which naturally shows that the verification result is accurate and reliable.

It is clear to those skilled in the art that in the cell experiments, the effect of the drug is only related to the drug concentration, not to the total amount of the drug, and meanwhile, the drug concentration at the cell level cannot be converted to the concentration in the animal experiments, so that the cell level experiments can only verify that the drug combination of the cepharanthine and epirubicin has no significant difference compared with the epirubicin used alone. Of course, the validity and reliability of the pharmaceutical composition of the present invention are also fully demonstrated by this validation. And the effect of the pharmaceutical composition is further verified by a nude mouse experiment, and only one tumor cell is researched.

Experimental example 2: nude mouse transplantation tumor model experiment (taking transplantation of human breast cancer MDA-MB-231 cells as an example only)

In the course of this experiment, the nude mice were injected intraperitoneally with various drugs for comparison (i.e., not intravenously).

1. Selection of nude mice

24 nude mice without specific pathogenic microorganisms (SPF) of 6-8 weeks old are taken, the weight range is 20 +/-2 g, and the nude mice are fed in the environment of room temperature (20-23 ℃) with alternate day and night, and are free to drink water and eat.

2. Experimental groups and dosages (4 groups in total)

All nude mice were randomly divided into four groups of 6 mice each, and 1X 10 mice were inoculated to the lateral side of each right thigh⁷0.2mL of a suspension prepared from human breast cancer MDA-MB-231 cells (the culture of human breast cancer MDA-MB-231 cells is the same as the aforementioned "cell experiment"), and administration is started after tumor formation in the following manner.

(1) Control group: injecting normal saline into abdominal cavity with volume same as that of treatment group for 1 time/day;

(2) cepharanthine group: injecting cepharanthine into abdominal cavity at a dose of 12mg/kg weight of nude mouse for 1 time/day;

(3) epirubicin group: injecting epirubicin intraperitoneally at a dosage of 2mg/kg body weight of nude mice for 1 time/5 days (i.e. injecting 1 time every 4 days);

(4) pharmaceutical combination group of cepharanthine and epirubicin: injecting cepharanthine into abdominal cavity at a dose of 12mg/kg weight of nude mouse for 1 time/day; epirubicin was intraperitoneally injected on the same day as the first cepharanthine injection at a dose of 2mg/kg body weight in nude mice, 1 time every 4 days.

The dose of epirubicin, 0.16mg, is calculated by injecting epirubicin 1 times/5 days according to the median value of the ratio (0.03-0.04) of epirubicin in the drug combination of the present invention.

The doses of cepharanthine and epirubicin in each group referred to above are simply the doses of the drug itself. The concentration to be achieved when formulating an injection will be readily apparent to those skilled in the art and can be determined by routine experimentation, at most, without further elaboration.

3. Measuring and recording

(1) Measuring and recording the weight of each group of nude mice after every six days;

(2) measuring the length and width of the tumor by using a vernier caliper after every six days, and recording the volume of the tumor body part of each group of nude mice once; tumor volume was calculated according to the following formula:

tumor volume (mm)³⁾Length of tumor (mm) × square of tumor width (mm)²)

Results of the experiment referring to FIGS. 6 to 8, FIG. 6 shows photographs taken after the end of the experiment (36 days), and the ordinate in FIG. 7 shows the tumor volume of each group of nude mice; the abscissa represents a measurement time point; various line nodes represent various processing groups; the values at each point are the mean of the tumor volumes of each group (the upper and lower deviations are indicated by the "I" symbols in the figure). The ordinate in fig. 8 represents the average body weight of each group of nude mice; the abscissa represents a measurement time point; various line nodes represent various processing groups; the values at each point are the average body weight of each group.

As can be seen from fig. 7, the tumor volume of the cepharanthine-synergistic epirubicin group was significantly lower after 12 to 18 days of treatment than the groups using epirubicin alone and cepharanthine alone. The ". mark" in fig. 7 also indicates p < 0.01, and it can be also confirmed that the tumor volume using the cepharanthine-epirubicin group is less likely to be erroneous than the drug group and the control group alone, and the verification result is also accurate and reliable.

As can be seen from fig. 8, the body weight of mice in the cepharanthine-epirubicin synergy group was not significantly different from those in the other drug-alone and control groups, compared with the epirubicin-alone, cepharanthine-alone and control groups. n.s represents p > 0.05, namely, the weight of the cepharanthine-epirubicin group is considered to be different from that of the single drug group or the control group, so that the error probability is more than 0.05, and because p 0.05 is the maximum statistically acceptable error probability, the error probability of the cepharanthine-epirubicin group which is more than 0.05 cannot be considered to be different from that of the single drug group or the control group, namely, no significant difference is found.

The verification result recorded in fig. 7 proves that the drug combination of the cepharanthine and the epirubicin chemotherapy is indeed superior to the single epirubicin, the MDA-MB-231 cells of the human breast cancer can be inhibited and killed, the dosage can be reduced, the treatment period can be shortened, and the toxic and side effects can be reduced. Since no significant toxic side effects are seen in the description of FIG. 3, the results of the validation demonstrate that the drug combination of cepharanthine in combination with epirubicin chemotherapy does not have at least the significant toxic side effects of epirubicin alone.

It is clear to those skilled in the art that after the verification of the above cell experiments, it can be completely demonstrated that the combination of the drugs of the cepharanthine and epirubicin chemotherapy has the same effect (difference and minimal) on human breast cancer BT549 cells, human leukemia K562 cells, U937 cells, gastric cancer SGC-7901 cells, lung cancer a549 cells and esophageal cancer Eca109 cells.

The foregoing is merely an example of the present invention and common general knowledge in the art of designing and/or characterizing particular aspects and/or features is not described in any greater detail herein. It should be noted that, for those skilled in the art, without departing from the technical solution of the present invention, several variations and modifications can be made, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicability of the patent. The scope of the claims of the present application shall be determined by the contents of the claims, and the description of the embodiments and the like in the specification shall be used to explain the contents of the claims.

Claims

1. A chemotherapeutic drug combination is characterized by consisting of cepharanthine and epirubicin with the mass ratio of 0.97: 0.032.

2. The use of a chemotherapeutic combination according to claim 1 for the preparation of an anticancer drug, wherein said chemotherapeutic combination consists of cepharanthine and epirubicin.

3. The use of a chemotherapeutic combination according to claim 2 for the preparation of an anticancer drug for the treatment of triple negative breast cancer.