Pharmaceutical composition for treating liver cancer

Abstract

The invention discloses a pharmaceutical composition for treating liver cancer, and belongs to the technical field of medicines. The pharmaceutical composition contains an effective dose of icariin and kaempferol, and the mass ratio of icariin to kaempferol is (8-12): (45-55). Experimental results show that compared with independent administration of KAE and ICA, a CMPLX group shows that more necrotic tissue fragment structures, moderate expansion of interstitial blood vessels, hyperemia and bleeding exist in tumor cell clusters, which indicates that KAE and ICA have a certain synergistic effect and have a more obvious killing effect on tumor cells after combined use, so that tumor cell proliferation is inhibited. Therefore, the invention provides a new medicine for treating the liver cancer.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and in particular relates to a pharmaceutical composition for treating liver cancer. Background Art

[0002] The trend of liver cancer is becoming increasingly severe worldwide. According to the 2020 Global Liver Cancer Statistics, liver cancer is the sixth most commonly diagnosed cancer in the world and the third leading cause of death from cancer. Liver cancer can be divided into primary and secondary liver cancers. The incidence of primary liver cancer is the highest among all liver cancers, and hepatocellular carcinoma is the most common primary liver cancer. Its mortality rate in the late stage is very high (>90%), which is usually caused by factors such as viral hepatitis (hepatitis B or C), aflatoxin infection, alcoholic liver disease and non-alcoholic fatty liver disease / disease. Among the liver cancer patient population, patients are often diagnosed with advanced liver cancer, which often has a poor prognosis, and surgical resection and drug chemotherapy are the best treatment options at this time. Indeed, Sorafenib is the first-line drug for the treatment of liver cancer. It is a multikinase inhibitor that targets cell growth and angiogenesis, and directly or indirectly causes cancer cell apoptosis through signaling pathways such as HIF-1α / SLC7A11 and ERK. However, in the past two years, studies exploring the relationship between Sorafenib and liver cancer have pointed to the drug resistance of liver cancer cells: cDCBLD2 (a circular RNA), SMYD3 (histone methyltransferase SET and MYND domain containing 3), DPP9 (dipeptidyl peptidase 9), GSTZ1 (glutathione S-transferase zeta1) and other substances directly / indirectly induce drug resistance in liver cancer cells treated with Sorafenib. Therefore, it is urgent to find new methods and drugs for the treatment of liver cancer, and traditional Chinese medicine treatment is an important complementary alternative therapy. Biqiong Ren et al. found that curcumin isolated from the rhizome of the traditional Chinese medicine turmeric promoted apoptosis by causing HepG2 cells to remain in the DNA S phase, and significantly reduced the levels of HSP70, eHSP70 and TLR4 in HepG2 cells under heat tolerance. Similar characteristic components of traditional Chinese medicine such as angelica polysaccharides and andrographolide have shown good anti-liver cancer activity in previous clinical studies.

[0003] As of 2020, there are more than 60 species of Epimedium plants recognized worldwide. The main production area is East Asia with the highest genetic diversity. China is the main distribution center, and it has been used as medicine for more than 2,000 years. The main medicinal varieties are the dried leaves of Epimedium brevicornu Maxim., Epimedium sagittatum (Sieb. et Zucc.) Maxim., Epimedium pubescens Maxim. and Epimedium koreanum Nakai, which mainly have the functions of nourishing the liver and kidneys, and dispelling wind and dampness.

[0004] Based on previous studies, scholars have detected more than 260 components in Epimedium, including flavonoids, lignins, phenolic glycosides, phenylethanol glycosides and polysaccharides. Among them, icariin, noricariin, Epimedin C and other components have shown anti-liver cancer activity through various mechanisms. At the beginning of 2022, icariin soft capsules were approved for marketing in China, which is a major breakthrough in the modernization of traditional Chinese medicine in the field of liver cancer.

[0005] However, the bioavailability of the above-mentioned monomers is rarely taken into consideration. Perhaps there are other monomer components or complexes with anti-liver cancer effects that still have anti-liver cancer therapeutic effects. Summary of the invention

[0006] The present invention provides a pharmaceutical composition for treating liver cancer, wherein the pharmaceutical composition contains an effective dose of icariin and kaempferol, wherein the mass ratio of icariin to kaempferol is 8-12:45-55, and the chemical formula of icariin is C 33 H 40 O 15 , molecular weight is 676.66, CAS registration number is 489-32-7, the chemical formula of kaempferol is C 15 H 10 O6, molecular weight is 286.236, CAS registration number is 520-18-3.

[0007] Preferably, the mass ratio of icariin to kaempferol is 1:5.

[0008] The present invention also provides a product for treating liver cancer, wherein the product contains the above-mentioned pharmaceutical composition.

[0009] Preferably, the product is a medicine.

[0010] More preferably, the medicine further contains medically acceptable excipients.

[0011] More preferably, the dosage form of the drug is any one of tablets, capsules, pills, granules and powders.

[0012] More preferably, the tablet is any one of enteric-coated tablets, sugar-coated tablets, plain tablets, film-coated tablets, dispersible tablets, sustained-release tablets, and controlled-release tablets.

[0013] More preferably, each tablet contains 769.23 mg of kaempferol and 153.85 mg of icariin, and is taken once a day, one tablet at a time.

[0014] More preferably, the capsule is any one of a hard capsule, a soft capsule, an enteric-coated capsule, a sustained-release capsule, and a controlled-release capsule.

[0015] The present invention also provides the use of icariin and kaempferol in preparing a combined drug for treating solid tumors, wherein the solid tumor is any one of liver cancer, pancreatic cancer, intestinal cancer, uterine cancer, prostate cancer, gastric cancer and the like.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] The present invention found that compared with the administration of KAE and ICA alone, the CMPLX group showed more necrotic tissue fragment structures in its tumor cell clusters, and moderate dilation, congestion and bleeding of blood vessels in the interstitium, indicating that KAE and ICA have a certain synergistic effect, and their combined use has a more obvious killing effect on tumor cells, thereby inhibiting tumor cell proliferation. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The results are the changes in tumor volume and tumor inhibition rate of mice in each drug-treated group in Example 2.

[0019] Figure 2 These are the results of hematoxylin-eosin staining of tumor tissue sections of mice in each group in Example 2 (200×, 400×). DETAILED DESCRIPTION

[0020] Example 1

[0021] Epimedium, Epimedium pubescens, Epimedium sagittatum and Epimedium koreana were powdered respectively, 25 times volume of 50% ethanol was added to the Epimedium powder respectively, and the supernatant was concentrated by rotary evaporation after ultrasonic treatment (power 80%, frequency 60kHz), and the remaining liquid was freeze-dried to obtain freeze-dried powder of Epimedium extract.

[0022] Example 2

[0023] 1 Experimental animals and methods

[0024] Five SPF male ICR mice, weighing 17-18 g, and 48 BALB / c male mice, weighing 17-18 g, were purchased from Jinan Pengyue Laboratory Animal Breeding Co., Ltd., license number SCXK (Lu) 20220006, and laboratory animal quality certificate number No. 370726241101030187. The animals were kept in an environmentally controlled room (temperature 24 ± 2 ° C, relative humidity 35%-65%, 12 h day and night alternation environment) with free access to food and water. There were no known contaminants in the food or water that might interfere with the research results. The animals were adapted for one week before the experiment. At the beginning of the study, all animals were in good health, and all animal operations were carried out in accordance with the Guide for the Care and Use of Laboratory Animals.

[0025] Forty-eight BALB / c male mice were randomly divided into model group (MOD), positive drug group (CTX), low-dose medicinal extract group (EPL), high-dose medicinal extract group (EPH), low-dose complex administration group (CMPLXL), high-dose complex administration group (CMPLXH), kaempferol group (KAE), and icariin group (ICA), with 6 mice in each group.

[0026] H22 cells in logarithmic growth phase were cultured at 2×10 6 The cells were inoculated at a density of 0.2 mL into the peritoneal cavity of each ICR mouse. After about 7 days, when the abdominal cavity of the mouse was obviously swollen, ascites was extracted and 0.1 mL was injected into the axilla of each Babl / c mouse after hair removal.

[0027] After 3 days, when obvious lumps appeared in the armpits of mice, BABL / c mice were gavaged once a day for a total of 7 days:

[0028] MOD group, control treatment (0.5% CMC-Na solution, 0.2 mL / mouse) was given by oral gavage;

[0029] CTX group, treated with cyclophosphamide saline solution (3 mg / ml cyclophosphamide saline solution, 0.2 mL / mouse) via intraperitoneal injection;

[0030] The EPL group was given a control treatment by oral gavage (6.1 mg of Epimedium extract, 6.4 mg of Epimedium pubescens extract, 5.85 mg of Epimedium sagittatum extract, and 8.5 mg of Epimedium koreana extract mixed and dissolved in 2 ml of 0.5% CMC-Na solution, 0.2 mL / mouse);

[0031] The EPH group was given a control treatment by oral gavage (10.1 mg of Epimedium extract, 10.7 mg of Epimedium pubescens extract, 9.75 mg of Epimedium sagittatum extract, and 14.17 mg of Epimedium koreana extract mixed and dissolved in 2 ml of 0.5% CMC-Na solution, 0.2 mL / mouse);

[0032] CMPLXL group, the complex (KAE+ICA, 50 mg / kg+10 mg / kg) was administered by oral gavage;

[0033] CMPLXH group, the complex (KAE+ICA, 100 mg / kg+20 mg / kg) was administered by oral gavage;

[0034] KAE group, administered by oral gavage (KAE, 50 mg / kg);

[0035] The ICA group was given by oral gavage (ICA, 10 mg / kg).

[0036] During the experiment, all animals received standard diet and water, and were free to eat. The rats' feed and water intake, abnormal behavior, and adverse toxicity signs were observed every day. During the experiment, the length and width of the tumor were measured every day with a vernier caliper, and the volume of the subcutaneous solid tumor in mice was calculated according to formula (1):

[0037] Tumor volume / mm 3 = length × width 2 / twenty one)

[0038] Twelve hours after the last administration, the tumor tissues of mice in each group were removed under deep anesthesia on a sterile operating table, and the tumor inhibition rate was calculated according to formula (2):

[0039] Tumor inhibition rate % = (average tumor weight of mice in the model group - average tumor weight of mice in the treatment group) / average tumor weight of mice in the model group × 100 (2)

[0040] Part of the tumor was preserved in 4% paraformaldehyde for later use, and hematoxylin-eosin (HE) staining and sections were made according to the method of the literature [Study on the anti-tumor effect of Pleurotus geesteranus powder on H22 tumor-bearing mice], observed under an optical microscope (200×, 400×), and photographed and recorded. All animal operations were carried out in accordance with the Guide for the Care and Use of Laboratory Animals.

[0041] 2 Experimental results

[0042] To observe whether the drug can inhibit tumor growth, the volume of subcutaneous solid tumors was measured once a day. Figure 1 A found that in the early stage of intervention, compared with the model group, the growth trend of the positive control group was slower, and the differences between the other groups were small. When cancer cells multiplied rapidly and the tumor volume continued to expand, each treatment group had a certain effect of inhibiting the tumor volume in the body, among which the inhibitory effect of the CMPLX group was better than that of KAE and ICA alone, and the high mass concentration inhibitory effect was better.

[0043] The tumor weight of each group of mice was further weighed, and the tumor weight of each drug-treated group was smaller than that of the model group ( Figure 1 B), indicating that all types of administration of Epimedium have a certain degree of inhibitory effect on tumors in tumor-bearing mice. The inhibition rate was calculated by tumor. Figure 1 C. Table 1 shows that compared with the single administration of KAE 50 mg / kg and ICA 10 mg / kg, the tumor inhibition rate was significantly increased after the combined administration of the two, with a relative increase of 168.0% and 74.3%, respectively, and the difference was extremely significant.

[0044] Table 1 Tumor mass and inhibition rate of mice in each drug administration group

[0045]

[0046] Note: Compared with the model group, *** indicates P < 0.001, * indicates P < 0.05.

[0047] Hematoxylin-eosin staining showed that in the model group, a large number of tumor cells were densely arranged in the muscle tissue with irregular cell morphology. The tumor cells were arranged in nests and trabeculae, and no obvious necrotic tissue fragment structure was observed in the tumor cell clusters. The tumor cells had rich cytoplasm and were eosinophilic. The nuclei were of different sizes, highly atypical, and had a large number of pathological nuclear divisions. The blood vessels in the interstitium were slightly dilated, congested, and hemorrhagic.

[0048] Among the drug-treated groups, the CMPLX group showed more necrotic tissue fragment structures in the tumor cell clusters, and moderate dilation, congestion, and bleeding of blood vessels in the interstitium compared with KAE and ICA alone, indicating that it has a more obvious killing effect on tumor cells, thereby inhibiting tumor cell proliferation.

[0049] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A pharmaceutical composition for treating liver cancer, characterized in that: The pharmaceutical composition contains effective doses of icariin and kaempferol, and the mass ratio of icariin to kaempferol is 8-12:45-55.

2. The pharmaceutical composition according to claim 1, characterized in that The mass ratio of icariin to kaempferol is 1:

5.

3. A product for treating liver cancer, characterized in that: The product contains the pharmaceutical composition according to claim 1 or 2.

4. The product according to claim 3, characterized in that The product described is a drug.

5. The product according to claim 4, characterized in that The medicine also contains medically acceptable auxiliary materials.

6. The product according to claim 4 or 5, characterized in that The dosage form of the medicine is any one of tablets, capsules, pills, granules and powders.

7. The product according to claim 6, characterized in that The tablet is any one of enteric-coated tablets, sugar-coated tablets, plain tablets, film-coated tablets, dispersible tablets, sustained-release tablets and controlled-release tablets.

8. The product according to claim 7, characterized in that Each tablet contains 769.23 mg of kaempferol and 153.85 mg of icariin, and is taken once a day, one tablet at a time.

9. The product according to claim 6, characterized in that The capsule is any one of a hard capsule, a soft capsule, an enteric-coated capsule, a sustained-release capsule, and a controlled-release capsule.

10. Use of icariin and kaempferol in the preparation of a combined drug for treating liver cancer.

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