阅读说明：本技术 5-ht受体抑制剂和顺铂在制备治疗肝癌的药物中的应用 (Application of 5-HT receptor inhibitor and cisplatin in preparation of medicine for treating liver cancer ) 是由 赵雪梅 李宁 王桂玲 付琳琳 王琢 狄伟华 叶彦艳 于 2021-09-23 设计创作，主要内容包括：本发明提供了五羟色胺受体抑制剂(5-HT受体抑制剂)和顺铂(Cisplatin)在制备治疗肝癌的药物中的应用,属于生物医药技术领域。5-HT受体抑制剂与Cisplatin联合用药作用于RECQL4高表达的肝癌,可以产生协同效应,能够增强RECQL4高表达的肝癌对Cisplatin的敏感性,增强Cisplatin的抗肝癌效果及减弱肝癌对Cisplatin的耐药性。(The invention provides application of serotonin receptor inhibitor (5-HT receptor inhibitor) and Cisplatin (Cisplatin) in preparing a medicament for treating liver cancer, belonging to the technical field of biological medicines. The 5-HT receptor inhibitor and Cisplatin are combined to act on the RECQL4 high-expression liver cancer, can generate a synergistic effect, and can enhance the sensitivity of the RECQL4 high-expression liver cancer to the Cisplatin, enhance the anti-liver cancer effect of the Cisplatin and weaken the drug resistance of the liver cancer to the Cisplatin.)

1. Application of serotonin receptor inhibitor and cisplatin in preparing medicine for treating liver cancer is provided.

2. Application of serotonin receptor inhibitor in preparing medicine for weakening resistance of liver cancer to cisplatin is provided.

3. Application of serotonin receptor inhibitor in preparing medicine for reducing toxic and side effects of cisplatin in treating liver cancer is provided.

4. Application of serotonin receptor inhibitor and cisplatin in preparing medicine for inhibiting liver cancer cell proliferation is provided.

5. The use of any one of claims 1 to 4, wherein the liver cancer comprises RECQL4 high expression liver cancer.

6. A medicine for treating liver cancer comprises serotonin receptor inhibitor and cisplatin.

7. The medicament according to claim 6, wherein the serotonin receptor inhibitor comprises one or more of mexicarpine, N- [3- (2-dimethylamino) ethoxy-4-methoxyphenyl ] -2' -methyl-4 ' - (5-methyl-1, 2, 4-oxadiazol-3-yl) - (1,1' -biphenyl) -4-carboxamide, 5- ((4- (6-chlorothieno [2,3-d ] pyrimidin-4-ylamino) piperidin-1-yl) methyl) -2-fluorobenzonitrile monofumarate and aripiprazole.

8. The medicament as claimed in claim 6, wherein the mass ratio of the serotonin receptor inhibitor to the cisplatin in the medicament is (1.5-2.5) to 5.

9. The medicament of claim 6, wherein the dosage form of the medicament comprises an injectable formulation.

10. The medicament of claim 9, wherein the concentration of serotonin receptor inhibitor in the medicament is 1.5-2.5 mg/ml; the concentration of cisplatin in the medicine is 4.5-5.5 mg/ml.

Technical Field

The invention relates to the technical field of tumor treatment, in particular to application of a 5-HT receptor inhibitor and cisplatin in preparation of a medicament for treating liver cancer.

Background

Liver cancer is cancer with incidence and fatality rate in prostate in the global scope, and currently, chemotherapy is performed on liver cancer treatment drugs represented by platinum in clinic, but due to individual differences of liver cancer patients, chemotherapy effects of patients in the same pathological period are remarkably different. At present, through analysis of a Cancer Genome Atlas (TCGA), changes of four genes including Rad21, Rad54B, Rad21 and RECQL4 are found to be related to The size, clinical stage and prognosis of tumors in clinical liver Cancer samples, particularly in liver Cancer of stages III and IV, The changes of The four genes including Rad21, Rad54B, Rad21 and RECQL4 have high correlation with prognosis, and The Rad21, Rad54B, Rad21 and RECQL4 genes are highly expressed in liver Cancer samples with poor prognosis. The four genes Rad21, Rad54B, Rad21 and RECQL4 are all involved in DNA replication and damage repair and play a role in different stages of DNA double strand break repair. The RECQL4 gene has stronger repair capability, and the expression level of the RECQL4 gene is in positive correlation with other repair genes; patients with higher RECQL4 expression have poorer prognosis and are susceptible to many clinical DNA damaging chemotherapeutic drugs (e.g., Cisplatin, Cis) (Thomassen M, Tan Q, Kruse T. Gene expression method-analysis identities in branched Cancer patients [ J ]. Breast Cancer Res Tr,2009,113(2): 239-; with the advent of chemotherapy, resistance of patients to chemotherapeutic drugs has led to treatment failure.

Serotonin receptor inhibitors (5-HT receptor inhibitors) are commonly used antidepressant drugs in clinic at present, and are mainly used for treating moderate and mild depression in clinic. At present, no report is available on the treatment of liver cancer by combining a serotonin receptor inhibitor and a chemotherapeutic drug.

Disclosure of Invention

The invention aims to provide application of a 5-HT receptor inhibitor and Cisplatin in preparation of a medicament for treating liver cancer, wherein the 5-HT receptor inhibitor can enhance the anti-liver cancer effect of Cisplatin and weaken the drug resistance of liver cancer to Cisplatin.

In order to achieve the above object, the present invention provides the following technical solutions:

the invention provides application of a serotonin receptor inhibitor and cisplatin in preparing a medicament for treating liver cancer.

The invention also provides application of the serotonin receptor inhibitor in preparing a medicine for weakening the drug resistance of liver cancer to cisplatin.

The invention also provides application of the serotonin receptor inhibitor in preparing a medicament for reducing the toxic and side effects of cisplatin in treating liver cancer.

The invention also provides application of the serotonin receptor inhibitor and the cisplatin in preparing the medicine for inhibiting the proliferation of the liver cancer cells.

Preferably, the liver cancer comprises a RECQL4 high-expression liver cancer.

The invention provides a medicine for treating liver cancer, which comprises a serotonin receptor inhibitor and cisplatin.

Preferably, the serotonin receptor inhibitor comprises one or more of mexicamine, N- [3- (2-dimethylamino) ethoxy-4-methoxyphenyl ] -2' -methyl-4 ' - (5-methyl-1, 2, 4-oxadiazol-3-yl) - (1,1' -biphenyl) -4-carboxamide, 5- ((4- (6-chlorothiophene [2,3-d ] pyrimidin-4-ylamino) piperidin-1-yl) methyl) -2-fluorobenzonitrile monofumarate and aripiprazole.

Preferably, the mass ratio of the serotonin receptor inhibitor to the cisplatin in the medicine is (1.5-2.5) to 5.

Preferably, the dosage form of the medicament comprises an injection.

Preferably, the concentration of the serotonin receptor inhibitor in the medicament is 1.5-2.5 mg/ml; the concentration of cisplatin in the medicine is 4.5-5.5 mg/ml.

The invention provides application of a serotonin receptor inhibitor and cisplatin in preparing a medicament for treating liver cancer. The 5-HT receptor inhibitor and Cisplatin are combined to act on the RECQL4 high-expression liver cancer, can generate a synergistic effect, can enhance the sensitivity of the RECQL4 high-expression liver cancer to the Cisplatin, enhance the anti-liver cancer effect of the Cisplatin, weaken the drug resistance of the liver cancer to the Cisplatin and weaken the toxicity of cis-platinum.

Drawings

FIG. 1 is a dose-effect relationship of PRX, Cis, PRX + Cis in inhibiting SNU398 cell proliferation;

FIG. 2 is a graph of Fa-CI for combined inhibition of SNU398 cell proliferation by PRX + Cis (. about.P < 0.05;. about.P < 0.01);

FIG. 3 is a dose-effect relationship of APZ, Cis, APZ + Cis for inhibiting SNU398 cell proliferation;

FIG. 4 is a Fa-CI plot of the combination of APZ + Cis inhibiting SNU398 cell proliferation (. about.P0.05;. about.P0.01);

FIG. 5 is a dose-effect relationship of MET, Cis, MET + Cis in inhibiting SNU398 cell proliferation;

FIG. 6 is a Fa-CI plot of the combination of MET + Cis inhibiting SNU398 cell proliferation (. beta.. P < 0.05;. beta.. P < 0.01);

FIG. 7 is a dose-effect relationship of SB, Cis, SB + Cis to inhibit SNU398 cell proliferation;

FIG. 8 is a Fa-CI plot of the combination of SB + Cis inhibiting SNU398 cell proliferation (P < 0.05;. P < 0.01);

FIG. 9 shows the tumor suppression ratios of nude mice by different administration groups in example 5 (.: P < 0.05;: P < 0.01);

FIG. 10 is a survival curve of nude mice of different administration groups in example 5.

Detailed Description

The invention provides application of a serotonin receptor inhibitor and cisplatin in preparing a medicament for treating liver cancer.

The invention provides application of a serotonin receptor inhibitor in preparing a medicine for weakening the drug resistance of liver cancer to cisplatin.

The invention provides application of a serotonin receptor inhibitor in preparing a medicament for reducing toxic and side effects of cisplatin in treating liver cancer.

The invention provides application of a serotonin receptor inhibitor and cisplatin in preparation of a medicament for inhibiting liver cancer cell proliferation.

In the present invention, the serotonin receptor inhibitor preferably includes one or more of Methiothepin (methohexine maleate), N- [3- (2-dimethylamino) ethoxy-4-methoxyphenyl ] -2' -methyl-4 ' - (5-methyl-1, 2, 4-oxadiazol-3-yl) - (1,1' -biphenyl) -4-carboxamide (SB 216641 hydrochloride), 5- ((4- (6-chlorothiophene [2,3-d ] pyrimidin-4-ylamino) piperidin-1-yl) methyl) -2-fluorobenzonitrile monofumarate (PRX-08066Maleic acid) and Aripiprazole (Aripiprazole). In the present invention, the Methothopepin maleate, SB 216641 hydrochloride, PRX-08066Maleic acid and Aripiprazole are conventionally commercially available.

In the present invention, the liver cancer preferably includes a RECQL4 high expression liver cancer.

In the present invention, the dosage form of the drug preferably includes an injection; the injection is preferably an intraperitoneal injection or an intravenous injection.

In the invention, the mass ratio of the serotonin receptor inhibitor to the cisplatin in the medicine is preferably (1.5-2.5) to 5. In the invention, the concentration of the serotonin receptor inhibitor in the medicament is preferably 1.5-2.5 mg/ml; the concentration of the cisplatin in the medicine is preferably 4.5-5.5 mg/ml. In the present invention, the solvent used for the drug is preferably sterile physiological saline or sterile PBS.

The invention also provides a medicament for treating liver cancer, which comprises serotonin receptor inhibitor and cisplatin.

The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The sources of the materials in the examples of the invention are as follows:

LO2 (human normal hepatocytes): purchased from Nanjing Kai Biotech development Ltd. SNU398 (human liver cancer cells): purchased from Guangzhou Saku Biotechnology, Inc. SB 216641 hydrochloride (Selective h5-HT1B antaconist, SB) and Methothepin maleate (5-HT Receptors, Non-Selective, MET) were purchased from TOCRTS; PRX-08066Maleic acid (5-HT2B receivers 1antagonist, abbreviated as PRX), Aripiprazole (a high-affinity 5-HT receiver partial aginst, abbreviated as APZ) was purchased from Selleck. Cisplatin (Cis for short) is purchased from Solambio.

Example 1 inhibition of cells by Cis and PRX alone and in combination

1. Cell plate

Selecting LO2 cells and SNU398 cells in logarithmic growth phase, digesting with 0.25% trypsin, centrifuging, collecting cells, counting cells, and diluting cells to 5 × 10³Each per ml, 3 parallel wells per group, 100. mu.l per well of cell suspension were seeded in 96-well plates at 37 ℃ in 5% CO₂After culturing in an incubator for 24h, observing the adherence condition of the cells under a microscope.

2. Combination drug

Grouping experiments:

(1) blank group: adding only culture medium without adding cells;

(2) DMSO negative group: no medicine is added;

(3) the single medicine group: 10 concentration levels (see Table 1) were set with a 2-fold relationship between adjacent concentrations.

(4) A combination of drugs: according to the general principle of Chou-Talalay combined exponential experiment design, 10 concentration levels are set for the medicine, and PRX and Cis are used in a combined way under different concentrations, namely the level 1: concentration of PRX 1+ concentration of Cis 1; level 2: concentration of PRX 2+ concentration of Cis 2; in turn to level 10: concentration of PRX 10+ concentration of Cis 10. (see Table 1).

TABLE 1 Effect of Cis and PRX alone and in combination on cell proliferation

And observing the cells in the 96-well plate under a microscope, wherein the cells are well attached to the wall and are full in shape, and the cells are ready to be added with medicine. Ultraviolet irradiating on a clean bench for 30min, circulating and ventilating for 15min, taking out the prepared drug mother liquor from a refrigerator, spraying 75% alcohol, and placing in a clean bench. And calculating and configuring the medicine according to the set medicine concentration. First, separate drug groups (PRX, Cis) were prepared, each at 10 concentrations, and Cis and PRX were combined. The 96-well plate was removed from the incubator, the original medium in the well was aspirated by a syringe, and different concentrations of each drug were added to the 96-well plate in separate drug wells, 100. mu.l per well. When the Cis and the PRX are used together, the volume ratio of the two medicines is 1:1, the total medicine amount in each hole is still 100 mu l, namely, the concentration of a single medicine is enlarged by one time and is respectively added by 50 mu l, and the corresponding concentration is obtained after dilution by multiple times as shown in the table 1.

The 96-well plate was removed from the cell incubator, the original medium was aspirated from each well using a syringe, and a fresh drug-containing medium was added to each well in an amount of 100. mu.l, and the 96-well plate was placed in the cell incubator to continue the culture. Aseptic operation is carried out in the dosing process to prevent cell pollution. And after 24h and 48h of drug intervention, respectively observing the cell morphology, and photographing for storage. After the medicine intervention is carried out for 48 hours, the medicine-containing culture medium is discarded, a certain amount of 10% CCK-8 working solution is prepared, 100 mu l of working solution is added into each hole, after the incubation is carried out for 2-4 hours in a cell incubator, the OD value of each hole under the wavelength of 450nm is measured by using a multifunctional microplate reader, the data is stored, and the data processing is carried out. The data is substituted into the following equation:

cell Viability (OD)_{Medicine using set}-OD_{Blank group})/(OD_{Negative group}-OD_{Blank group})×100％；

OD_{Medicine using set}Is the final solution light absorption value of the cell hole measured after adding the compound to be measured for co-culture;

OD_{negative group}Is the final solution absorbance value of the cell pore measured after adding complete culture medium for co-culture;

OD_{blank group}Is the final solution absorbance value of a blank well of medium without cells and test compound;

taking the concentration of the drug as the abscissa and the survival rate of the cells as the ordinate, GraphPad Prism 6.0 was used for data processing to obtain the IC corresponding to each drug for each cell₅₀The value, 50% induction of tumor cell death by a concentration of compound called the median inhibitory concentration, is a measure of the ability of a drug to interfere with tumor cell proliferation, IC₅₀The lower the value, the stronger the drug intervention ability, indicating a cell pairThe more sensitive the drug.

3. Chou-Talalay equation and determination of combined drug effect

According to the Chou-Talalay equation, fa/fu is [ D/Dm ] m, (D is the drug concentration, Dm is the median effect concentration, namely the drug concentration when fa is 0.5), taking the logarithm of two sides of the linear equation: y ═ mX + a (where Y ═ log [ fa/fu ], X ═ logD, m is the slope of the equation, and a is the equation intercept). The corresponding Dm (logDm ═ a/m) of the two drugs used alone or in combination is calculated respectively, and then the effect relationship of the two drugs used in combination is calculated according to the dose-effect formula in the combined index method, namely D1/Dx1+ D2/Dx2+ alpha D1D2/Dx1Dx 2. (D1 and D2 indicate the concentrations of the two drugs when the drug effect X is achieved, Dx1 and Dx2 are respectively used when the two drugs are used independently, the interaction of the two drugs is that CI <1 indicates the synergistic effect when the two drugs are used together, CI ═ 1 indicates the additive effect when the two drugs are used together, and CI >1 indicates the antagonistic effect when the two drugs are used together.)

Drug intervention was performed on SNU398 by setting PRX, Cis, and PRX + Cis groups using LO2 cells as a control, and the results are shown in fig. 1 and 2. The results in FIG. 1 show that: the Cis group, the PRX group and the PRX + Cis group have inhibition effect on the proliferation of the SNU398 cells, the inhibition rate of the cell proliferation is increased along with the increase of the administration concentration, and the dosage-effect relationship is realized; compared with the Cis group, the inhibition rate of the PRX + Cis group to the SNU-398 cells is obviously improved, and the statistical difference is significant (P is less than 0.01); when the Cis group and the PRX + Cis group reach the same inhibition rate, the PRX + Cis group reduces the dosage of Cis and enhances the sensitivity of SNU398 to Cis. The results in FIG. 2 show that when the PRX + Cis combination is used, the CI value is less than 1, the synergistic effect is shown under different concentrations, and the synergistic effect of the PRX + Cis combination is enhanced along with the increase of the concentration of the combination.

EXAMPLE 2 cellular inhibition by Cis and APZ alone and in combination

See table 2 for drug grouping and example 1 for the rest of the procedure.

TABLE 2 Effect of Cis and APZ alone and in combination on cell proliferation

Drug intervention was performed on SNU398 by setting APZ group, Cis group, and APZ + Cis group using LO2 cells as a control. As shown in FIG. 3, Cis group, APZ group and APZ + Cis group all have inhibitory effects on the proliferation of SNU398 cells, and the inhibition rate of cell proliferation is increased along with the increase of the drug concentration, and has a dose-effect relationship; at medium and low concentration, compared with the Cis group, the inhibition rate of the SNU-398 cells by the APZ + Cis group is higher than that of the Cis group, and the statistical difference (P <0.05) exists, which indicates that the sensitivity of the SNU398 to Cis is improved by the combined medication of the APZ + Cis group and the Cis group; with the increase of the drug concentration, the inhibition rates of the Cis group and the APZ + Cis group to the SNU398 have no obvious difference; in FIG. 4, when APZ + Cis group is used in combination at low concentration, the CI value is less than 1, and the synergistic effect is shown; as the concentration of the APZ + Cis group combination is increased, the CI value is larger than 1, and antagonism is presented.

Example 3 inhibition of cells by Cis and MET alone and in combination

See table 3 for drug grouping and example 1 for the rest of the procedure.

TABLE 3 Effect of Cis and MET alone and in combination on cell proliferation

With LO2 cells as control, the MET group, Cis group, and MET + Cis group were set for SNU398, and drug intervention was performed. As shown in FIG. 5, Cis group, MET group and MET + Cis group all have inhibitory effects on the proliferation of SNU398 cells, and the inhibition rate of cell proliferation is increased along with the increase of the administration concentration, and has a dose-effect relationship; in a certain concentration range, the inhibition rate of the MET + Cis group to the SNU398 is obviously higher than that of the Cis group, and the statistical difference is significant (P < 0.01); as shown in FIG. 6, when the combination of MET + Cis group at low concentration was administered, the CI value >1 exhibited antagonism, and when the combination of MET + Cis group at high concentration was administered, the CI value <1 exhibited synergy.

Example 4 inhibition of cells by Cis and SB alone and in combination

See table 4 for drug grouping and example 1 for the rest of the procedure.

TABLE 4 Effect of Cis and SB on cell proliferation alone and in combination

Drug intervention was performed on SNU398 by setting SB group, Cis group, and SB + Cis group using LO2 cells as a control. As shown in FIG. 7, the Cis group, the SB group and the SB + Cis group all have inhibitory effects on the proliferation of SNU398 cells, and the inhibition rate of cell proliferation increases with the increase of the drug concentration, and has a dose-effect relationship; in a certain concentration range, compared with the Cis group, the inhibition rate of the SB + Cis group to the SNU398 is obviously improved, and the statistical difference is significant (P < 0.01); as shown in FIG. 8, when the low concentration combination of SB + Cis group was used, the CI value was >1, and when the high concentration combination of SB + Cis group was used, the CI value was <1, showing a synergistic effect.

Example 5 pharmacodynamic assay of APZ and Cis combination for nude mouse xenograft tumor models

(1) The method comprises the following operation steps:

after the BALB/C-Nu nude mice were cultured in SPF environment for 1 week, 40 nude mice were individually weighed and labeled. Sterile PBS and Matrigel are prepared in a volume ratio of 1:1 on ice and mixed evenly, SNU398 cells are fixed into cell suspension and injected into right forelimb armpit of a nude mouse, 200 mu l of mixed solution is injected into the nude mouse of Blank group (PBS: Matrigel ═ 1:1, volume ratio), and 7 × 10 of nude mice of experimental group are injected into the nude mouse of experimental group⁶200 μ l of liver cancer cell SNU 398. After the nude mice are injected with SNU398 cells subcutaneously, the growth condition of the tumor is observed every day, and the volume of the tumor to be grown in the nude mice is about 100-150 mm³In the process, according to the size of the tumor and the weight of the nude mice, the nude mice with over-large or over-small tumor are removed, and are randomly grouped for intraperitoneal injection administration. The experimental groups were divided into 5 groups: (1) cis drug group: the dosage is 25mg/kg, and the medicine is taken once every five days; (2) APZ group: the dosage is 10mg/kg, and the medicine is taken once every two days; (3) cis and APZ combination: administered once every five days at a dose of Cis (25mg/kg) + APZ (10 mg/kg); (4) DMSO control group: injecting DMSO solvent every timeOnce every two days; (5) blank group.

Auxiliary materials of the medicine: preparing the medicine by using sterile PBS (pH7.2); configuration of cissplatin: 50mg of Cis were added to 50. mu.l DMSO and 9950. mu.l PBS was added to prepare a solution of 50mg/10ml, which was dispensed into 10 EP tubes of 1.5ml and frozen at-20 ℃ for use. The dosage of each tumor-bearing mouse is as follows: 100 μ l/20 g. Configuration of APZ: 20mg of APZ was used to prepare a 10ml solution, otherwise the procedure was as for Cis. The dosage of each tumor-bearing mouse is as follows: 100 μ l/20 g.

Observing whether the mental state of the mouse is normal after injecting the medicine, recording death time by taking administration days as a unit, drawing a survival cycle curve and evaluating the survival cycle; observing and measuring the weight of the nude mice every other day, measuring the long diameter and the short diameter of the tumor by using a vernier caliper, and recording and analyzing. After the experiment is finished, cervical vertebrae are removed, animals are sacrificed and dissected, tumors are stripped, the weight of the nude mice and the tumor weight are weighed, the heart, the liver, the spleen, the lung and the kidney are taken, then the weighing is carried out, the organ coefficients of each group are respectively calculated, and the results are statistically processed. The analytical data are expressed as Mean ± standard deviation (Mean ± s.e.m), comparative analysis of data between groups using One-Way analysis of variance (One Way ANOVA), comparison between two groups using student-t test, statistical analysis and plotting using GraphPad Prism 6.0 statistical software.

(2) As a result: tumor inhibition effect of combined medicine

After the experiment, the nude mice were sacrificed by removing the cervical vertebrae, the nude mice were dissected to remove the tumor, and the tumor was weighed. The tumor inhibition rate was calculated by the following formula: the inhibition ratio (%) (tumor weight of control group-tumor weight of experimental group)/tumor mass of control group × 100%. As shown in FIG. 9 and Table 5, compared with the DMSO control group, the APZ group, the Cis group and the APZ + Cis group have an inhibitory effect on the growth of tumors in mice, wherein the inhibition rate of the APZ + Cis group on the tumors is highest. The inhibition rates of APZ group, Cis group and APZ + Cis group on tumors are respectively 25.02%, 82.33% and 91.23%.

TABLE 5 inhibition of tumor growth by APZ, Cis, APZ and Cis

Compared to DMSO control group,: p < 0.05; **: p < 0.01.

(3) As a result: survival Curve analysis in nude mice

The survival cycles of nude mice of different drug groups were evaluated and survival cycle curves were plotted, as shown in fig. 10, it can be seen that the survival cycles of the nude mice of Cis group, APZ + Cis group were all prolonged compared to the DMSO control group, wherein the survival cycle of the nude mice of APZ + Cis group was the longest.

(4) As a result: analysis of organ coefficients of nude mice

When tumor model mice were treated in the above experiment, the heart, liver, spleen, lung and kidney were removed. Each organ was then weighed and the weight recorded. Organ coefficients are calculated according to the formula: the organ coefficient is the organ weight/body weight × 100%.

After each group of BALB/N-Cu nude mice is dissected, organs are taken and weighed, the statistics and analysis of main organ coefficients are carried out, the results are shown in Table 6, compared with BALB/N-Cu nude mice of a DMSO control group, the organ coefficients of the lungs of a Cis group, an APZ + Cis group are respectively increased to different degrees, and the statistical difference is significant (P is less than 0.01); the organ coefficient of Cis group kidney is increased, and the statistical difference is significant (P < 0.01); the organ coefficient of the Cis group heart is increased, and the statistical difference is obvious (P < 0.01); compared with the mouse BALB/N-Cu of the Cis group, the lung organ coefficients of the APZ group and the APZ + Cis group are higher than those of the Cis group, and the statistical difference is significant (P is less than 0.01); the organ coefficient of the spleen of the APZ group is higher than that of the Cis group, and the statistical difference exists (P is less than 0.01); the organ coefficients of the APZ group and the APZ + Cis group are lower than those of the Cis group, and the statistical difference is significant (P is less than 0.05); compared with organ coefficients of liver and spleen of the DMSO control group, the Cis group, the APZ + Cis group and the DMSO control group, no statistical difference exists.

TABLE 6 BALB/C-Nu mouse organ coefficient analysis

Each group compared to DMSO control group,: p < 0.05; **: p < 0.01.

Organ coefficient analysis shows that organ coefficients of kidneys, hearts and lungs of the Cis group are higher than those of organs of nude mice of the DMSO group, and the Cis group has large toxic and side effects and can cause complications. The inhibition rate of the APZ + Cis combined drug group on tumors is highest, and organ coefficient analysis shows that compared with the Cis group, organ coefficients of the kidney, the heart and the lung of the APZ + Cis group are all smaller than those of the Cis group, so that the APZ can weaken toxic and side effects of the Cis and enhance the sensitivity of tumors to the Cis to a certain extent.

The side effects of cissplatin are manifested as loss of appetite, nausea, vomiting, diarrhea, etc. Usually appear 1-2 hours after injection, last 4-6 hours or more, and disappear after 2-3 days. This response is dose dependent. Cisplatin has prominent nephrotoxicity, mainly manifested as renal transient injury, including damage to tubular epithelial cells. It is characterized by impairment of hematuria and renal function, increase of serum creatinine and reduction of clearance rate.

The above examples show that the 5-HT receptor inhibitor can enhance the anti-liver cancer effect of Cisplatin and reduce the toxic and side effects of Cisplatin, and provides a new treatment idea and theoretical experimental basis for clinically treating liver cancer.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.