Application of combination of punicalagin and oxaliplatin in preparation of medicine for treating gastric cancer ovarian metastasis

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to application of a combination of punicalagin and a chemotherapeutic drug oxaliplatin (OXA) in preparation of a drug for treating gastric cancer ovarian metastasis. Researches show that in various animal models of gastric cancer ovarian metastasis of mice, single treatment of oxaliplatin cannot inhibit tumor growth, punicalagin shows a certain effect of inhibiting tumor growth, and punicalagin and oxaliplatin are combined for medication to show significant synergistic interaction and significantly inhibit tumor growth. The pharmaceutical composition is simple in component and can be used for treating gastric cancer ovarian metastasis, especially for patients insensitive to oxaliplatin, and punicalagin and oxaliplatin are both marketed drugs, can be quickly developed and applied clinically and have good popularization and application prospects.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to the use of a combination of punicalagin and oxaliplatin in the preparation of a therapeutic drug for gastric cancer ovarian metastasis. Background Art

[0002] Gastric cancer is a common malignant tumor worldwide, characterized by insidious onset and high metastasis rate. More than half of patients have developed distant metastasis at the time of initial diagnosis, including lymph nodes, liver, lungs, bones, peritoneum and ovaries. Among them, ovarian metastasis is a special type of metastasis of gastric cancer, which is common in young women and has common symptoms such as abdominal pain, abdominal distension, irregular vaginal bleeding or increased leucorrhea, which is seriously life-threatening. The prognosis of patients with ovarian metastasis of gastric cancer is worse than that of ovarian metastatic tumors from other digestive tract sources, with a median survival time of only 7-14 months.

[0003] Currently, the treatment of gastric cancer with ovarian metastasis remains controversial internationally. The probability of surgical resection of isolated ovarian metastasis is only 17% to 25%, and most patients require chemotherapy. Currently, the majority of chemotherapy regimens are oxaliplatin-based. Oxaliplatin (CAS No: 61825-94-3), also known as oxaliplatin or oxaliplatin, is a platinum derivative and a third-generation platinum anti-tumor compound following cisplatin and carboplatin. It is also the only cheloplatin-based drug with significant activity against colorectal cancer. It acts by generating alkylated conjugates that bind to DNA, forming intra- and interstrand crosslinks that inhibit DNA synthesis and replication. It also exhibits antiproliferative effects in ovarian cancer and melanoma cell lines. Oxaliplatin has demonstrated broad in vitro cytotoxicity and in vivo antitumor activity in various tumor models, including gastric and ovarian cancers. It is often combined with anthracyclines, taxol, organic acids, and protein inhibitors to enhance its anticancer efficacy. However, for gastric cancer with ovarian metastasis, the survival benefit brought by oxaliplatin-based chemotherapy regimens is very limited, and many patients are insensitive to oxaliplatin. The clinical treatment of gastric cancer with ovarian metastasis still faces difficulties.

[0004] Drug combinations are a common clinical synergistic approach. For example, CN107669685A discloses that a three-drug combination of S-1a, paclitaxel, and apatinib has shown promising results for gastric cancer with ovarian metastasis. However, drug combination synergy is highly selective and can even lead to negative effects such as antagonism. Currently, there are relatively few drug combinations available for the treatment of gastric cancer with ovarian metastasis, making it difficult to meet clinical needs. Summary of the Invention

[0005] The present application aims to develop a new drug for treating gastric cancer ovarian metastasis. Through research, a simple drug combination for effectively treating gastric cancer ovarian metastasis is provided, i.e., a combination therapy of punicalagin and oxaliplatin, which has good application value for treating gastric cancer ovarian metastasis. According to the research of the present application, in various mouse gastric cancer ovarian metastasis animal models, oxaliplatin alone cannot inhibit tumor growth, while punicalagin shows a certain inhibitory effect on tumor growth, and punicalagin combined with oxaliplatin shows significant synergistic effect.

[0006] The present application aims to provide the application of punicalagin in preparing a drug for treating or inhibiting gastric cancer metastasis, especially gastric cancer ovarian metastasis.

[0007] The present application aims to provide the application of punicalagin in preparing a drug for treating or inhibiting gastric cancer metastasis, especially gastric cancer ovarian metastasis.

[0008] The present application aims to provide the application of punicalagin in preparing a drug for treating or inhibiting gastric cancer metastasis, especially gastric cancer ovarian metastasis.

[0009] The above-mentioned purposes of the present application are achieved by the following technical solutions.

[0010] The present application provides a combined medication scheme of punicalagin and the chemotherapeutic drug oxaliplatin. According to the research, in various mouse gastric cancer ovarian metastasis animal models, oxaliplatin alone cannot inhibit tumor growth, while punicalagin shows a certain inhibitory effect on tumor growth, and punicalagin can improve the efficacy of oxaliplatin, and the combination of the two can significantly inhibit tumor growth. The drug combination has simple components and can be used for treating gastric cancer ovarian metastasis, and has important application value for treating gastric cancer ovarian metastasis.

[0011] Therefore, specifically, the present application provides and claims the following scheme: the application of punicalagin in preparing a drug for treating or inhibiting gastric cancer metastasis.

[0012] Preferably, it refers to the application of punicalagin in preparing a drug for treating or inhibiting gastric cancer ovarian metastasis.

[0013] The application of punicalagin in preparing a synergistic agent for oxaliplatin in treating or inhibiting gastric cancer metastasis.

[0014] Preferably, it refers to the application of punicalagin in preparing a synergistic agent for oxaliplatin in treating or inhibiting gastric cancer ovarian metastasis.

[0015] More specifically, the present application provides a drug combination comprising punicalagin and oxaliplatin.

[0016] and the application of the drug combination in preparing a drug for treating gastric cancer ovarian metastasis.

[0017] Preferably, it refers to the use of the drug combination in the preparation of a therapeutic drug for gastric cancer ovarian metastasis.

[0018] In the above-mentioned combination drug regimen, the main active ingredients of the drug combination are punicalagin and oxaliplatin.

[0019] Preferably, the active ingredients of the drug combination are only: punicalagin and oxaliplatin.

[0020] Preferably, the pharmaceutical combination further comprises pharmaceutically acceptable excipients, such as carriers, excipients, and / or vehicles, and can be formulated into different pharmaceutical dosage forms as needed.

[0021] Punicalagin (CAS No. 65995-63-3), also known as punicalagin, belongs to the ellagitannin subclass of hydrolyzable tannins. Punicalagin is abundant in pomegranate juice, fruit, peel, and bark. It is also found at high levels in pink rock rose, tropical almond, yellowwood, and Terminalia chebula. In cancer research, punicalagin has been shown to be effective in treating oral, gastric, intestinal, cervical, breast, ovarian, lung, prostate, and thyroid cancers.

[0022] However, the use of punicalagin in the treatment of gastric cancer with ovarian metastasis has not been reported, especially for patients with gastric cancer with ovarian metastasis who have poor chemotherapy efficacy, and the anticancer effect of punicalagin is unknown. The present invention innovatively applies it to enhance the therapeutic effect of oxaliplatin on gastric cancer with ovarian metastasis, achieving a significant improvement in efficacy. This invention not only provides a new treatment option for patients with gastric cancer with ovarian metastasis, but also provides a new application direction for punicalagin, and also plays an important role in the development of drug value.

[0023] The present invention has the following beneficial effects:

[0024] (1) The present invention provides a drug combination of punicalagin and oxaliplatin, which is used to treat ovarian metastasis of gastric cancer. By combining these two drugs, tumor growth is significantly inhibited.

[0025] (2) The present invention provides a significantly effective and simple combination of therapeutic drugs for the treatment of ovarian metastasis of gastric cancer. Moreover, all of the drugs are already on the market and can be quickly developed and applied in clinical practice, with good prospects for promotion and application. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is the construction of a mouse gastric cancer ovarian metastasis model. Figure A shows the location of the mouse abdomen, Figure B shows the location of the ovary, Figure C shows the tumor inoculation, Figure D shows the in vivo imaging, Figure E shows the tumor in the ovary, and Figure F shows the HE staining of the pathological section.

[0027] Figure 2Therapeutic effect of punicalagin and oxaliplatin in NUGC4 mouse model of gastric cancer ovarian metastasis, wherein, A is the live imaging results of NUGC4 cell line ovarian metastasis model, B is the live imaging statistics, C is the tumor photos of the ovarian site, D is the tumor weight statistics. (Control: control group, OXA: only given oxaliplatin treatment of treatment group 1, Punicalagin: only given punicalagin treatment of treatment group 2, OXA+Punicalagin: given oxaliplatin and punicalagin combination therapy of treatment group 3, Week 1: one week of experiment, Week 3: three weeks of experiment, Total flux: total flux, Tumor weight: tumor weight, ns: no statistical difference, *: P<0.05, **: P<0.01, ***: P<0.001).

[0028] Figure 3 Therapeutic effect of punicalagin and oxaliplatin in NUGC4 mouse model of gastric cancer ovarian metastasis, wherein, A is the live imaging results of NUGC4 cell line ovarian metastasis model, B is the live imaging statistics, C is the tumor photos of the ovarian site, D is the tumor weight statistics. (Control: control group, OXA: only given oxaliplatin treatment of treatment group 1, Punicalagin: only given punicalagin treatment of treatment group 2, OXA+Punicalagin: given oxaliplatin and punicalagin combination therapy of treatment group 3, Week 1: one week of experiment, Week 3: three weeks of experiment, Total flux: total flux, Tumor weight: tumor weight, ns: no statistical difference, *: P<0.05, **: P<0.01, ***: P<0.001). DETAILED DESCRIPTION

[0029] The present application will be further described below in conjunction with the accompanying drawings and specific examples, but the examples do not make any form of limitation to the present application. Unless otherwise specified, the reagents, methods and devices used in the present application are conventional reagents, methods and devices in the art. Unless otherwise specified, the reagents and materials used in the examples are commercially available.

[0030] The technical solutions in the embodiments of the present application will be described clearly and completely below. Obviously, the described embodiments are only a part of the embodiments of the present application, not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor are within the protection scope of the present application.

[0031] Example 1 Construction of mouse model of gastric cancer ovarian metastasis

[0032] 1. Experimental materials

[0033] (1) Cells: Fluorescent dye-labeled gastric cancer cell lines MKN45 and NUGC4.

[0034] (2) Mice: Commercially available nude mice.

[0035] (3) Surgical instruments, anesthetic drugs, and disinfectants.

[0036] (4) In vivo imaging equipment.

[0037] (5) Clean bench.

[0038] (6) Insulated countertop.

[0039] 2. Experimental steps

[0040] (1) First, fluorescent dye-labeled gastric cancer cell lines MKN45 and NUGC4 were established. Cells in the logarithmic phase were collected to prepare cell suspensions. After disinfection, 50 μL of the cell suspension was injected subcutaneously into 5-week-old nude mice. The total number of cells was 1×10 6 After 2 weeks of tumor formation, the tumor tissue was cut into pieces and used for inoculation in the ovary.

[0041] (2) Perform intraperitoneal anesthesia on another 5-week-old nude mice. After the mice are in a deep anesthesia state, place them in a lateral position and place them flat on a clean work surface. After disinfection, make an opening on the back of the nude mouse thigh ( Figure 1 Middle A). Use forceps to reach into the nude mouse and find the ovary ( Figure 1 Figure B, red circle), inoculated about 1mm on the ovary 3 Size of the tumor tissue block (1) Figure 1 (C) After inoculation, the wound was sutured, disinfected, and placed on a warming table. Once the mouse was awake, it was returned to its cage.

[0042] 3. Experimental results

[0043] The growth of tumors was observed by in vivo imaging systems, such as Figure 1 As shown in Figure D, the luminescent dye shows a tumor growing in the ovary, indicating that the model was successfully established.

[0044] About 3 weeks after the modeling surgery, the mice can be euthanized and the ovarian metastatic tumors can be removed for photographing and weighing. Figure 1 As shown in Figure E. HE staining identified the tissue source (ovary), as shown in Figure 1 As shown in Figure F.

[0045] The results showed that this method was successful in constructing a mouse gastric cancer ovarian metastasis model.

[0046] Mice modeled according to this method can be divided into groups for treatment about one week after the modeling surgery. Example 2 Effect of Punicalagin and Oxaliplatin Combination on Ovarian Metastasis of Gastric Cancer in MKN45 Mice

[0047] 1. Experimental Materials

[0048] (1) Drugs: Commercially available punicalagin (MCE, HY-N0063) and oxaliplatin (MCE, HY-17371).

[0049] (2) Surgical instruments, anesthetic drugs, and disinfectants.

[0050] (3) In-vivo imaging equipment.

[0051] (4) The MKN45 mouse gastric cancer ovarian metastasis model was constructed according to the method of Example 1.

[0052] (5) Weighing equipment.

[0053] 2. Experimental steps (1) One week after modeling, the mice were divided into 4 groups, 6 in each group, and given:

[0054] Control group: given drug solvent (water) treatment,

[0055] Treatment group 1: 3 mg / kg of oxaliplatin was given.

[0056] Treatment group 2: 100 mg / kg of punicalagin was given.

[0057] Treatment group 3: 3 mg / kg oxaliplatin and 100 mg / kg punicalagin were given in combination.

[0058] Among them, oxaliplatin is taken once every three days for three times; punicalagin is taken once a day for 10 days.

[0059] (2) Monitor tumor growth every week using an in vivo imaging device.

[0060] (3) Two weeks after the start of treatment, the mice were euthanized, and the tumor tissues were removed, weighed, and photographed, and the differences between the groups were statistically analyzed.

[0061] 3. Experimental results

[0062] The results are as follows Figure 2 In vivo imaging was performed again at week 3 (as shown Figure 2 As shown in Figure A), Figure 2The statistical data in Figure B show that oxaliplatin has no significant inhibitory effect on tumor growth when used alone, and has a certain inhibitory effect on tumor growth when used alone. The two compounds have a significant inhibitory effect on tumor growth when used in combination, and there is a significant statistical difference compared with the use of oxaliplatin or punicalagin alone, indicating that the drug combination of punicalagin and oxaliplatin has more advantages in the treatment of ovarian metastasis of gastric cancer, and the two have played a significant synergistic role.

[0063] Nude mice were euthanized, and the ovarian tumors were dissected and photographed, weighed, and statistically analyzed. Figure 2 As shown in Figures CD, when oxaliplatin was used alone, the tumor size and weight were similar to those in the control group. When punicalagin was used alone, the tumor size and weight were slightly reduced compared to the control group. When the two compounds were used in combination, the tumor size and weight were significantly reduced compared to the control group. This further confirmed that Figure 2 The results are shown in Figure AB.

[0064] Example 3 Effect of Punicalagin and Oxaliplatin Combination on Ovarian Metastasis Growth of Gastric Cancer in NUGC4 Mice

[0065] 1. Experimental Materials

[0066] (1) Drugs: Commercially available punicalagin (MCE, HY-N0063) and oxaliplatin (MCE, HY-17371).

[0067] (2) Surgical instruments, anesthetic drugs, and disinfectants.

[0068] (3) In-vivo imaging equipment.

[0069] (4) The NUGC4 mouse gastric cancer ovarian metastasis model was constructed according to the method of Example 1.

[0070] (5) Weighing equipment

[0071] 2. Experimental steps (1) One week after modeling, the mice were divided into 4 groups, 6 in each group, and given:

[0072] Control group: given drug solvent (water) treatment,

[0073] Treatment group 1: 3 mg / kg of oxaliplatin was given.

[0074] Treatment group 2: 100 mg / kg of punicalagin was given.

[0075] Treatment group 3: 3 mg / kg oxaliplatin and 100 mg / kg punicalagin were given in combination.

[0076] Among them, oxaliplatin is taken once every three days for three times; punicalagin is taken once a day for 10 days.

[0077] (2) Monitor tumor growth every week using an in vivo imaging device.

[0078] (3) Two weeks after the start of treatment, the mice were euthanized, and the tumor tissues were removed, weighed, and photographed, and the differences between the groups were statistically analyzed.

[0079] 3. Experimental results

[0080] The results are as follows Figure 3 As shown, consistent with the results in Example 2, in vivo imaging was performed again at week 3 (as shown in FIG. Figure 3 As shown in Figure A), Figure 3 The statistical results in Figure B show that oxaliplatin had no significant inhibitory effect on tumor growth when used alone, and had a certain inhibitory effect on tumor growth when used alone. The two compounds had a significant inhibitory effect on tumor growth when used in combination, and there was a significant statistical difference compared with oxaliplatin or punicalagin used alone, indicating that the drug combination of punicalagin and oxaliplatin has more advantages in the treatment of ovarian metastasis of gastric cancer, and the two have played a significant synergistic role.

[0081] Nude mice were euthanized, and the ovarian tumors were dissected and photographed, weighed, and statistically analyzed. Figure 3 As shown in Figures CD, when oxaliplatin was used alone, the tumor size and weight were similar to those in the control group. When punicalagin was used alone, the tumor size and weight were slightly reduced compared to the control group. When the two compounds were used in combination, the tumor size and weight were significantly reduced compared to the control group. This further confirmed that Figure 3 The results are shown in Figure AB.

[0082] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be considered as equivalent replacement methods and are included in the scope of protection of the present invention.

Claims

1. Application of punicalagin in the preparation of drugs for treating or inhibiting gastric cancer metastasis.

2. Application of punicalagin in the preparation of drugs for treating or inhibiting ovarian metastasis of gastric cancer.

3. Application of punicalagin in the preparation of oxaliplatin synergist for treating or inhibiting gastric cancer metastasis.

4. Application of punicalagin in the preparation of oxaliplatin synergist for treating or inhibiting ovarian metastasis of gastric cancer.

5. A drug combination for treating or inhibiting gastric cancer metastasis, characterized in that: Including punicalagin and oxaliplatin.

6. The pharmaceutical combination according to claim 5, characterized in that The main active ingredients of the drug combination are punicalagin and oxaliplatin.

7. The pharmaceutical combination according to claim 5 or 6, characterized in that The active ingredients of the drug combination are only punicalagin and oxaliplatin.

8. The pharmaceutical combination according to claim 5 or 6, characterized in that The pharmaceutical combination further includes pharmaceutically acceptable excipients.

9. Use of the drug combination according to any one of claims 5 to 8 in the preparation of a drug for treating or inhibiting gastric cancer metastasis, wherein the drug combination comprises punicalagin and oxaliplatin.

10. The use according to claim 9, characterized in that The gastric cancer metastasis is gastric cancer ovarian metastasis.

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