阅读说明：本技术 雷洛昔芬与奥沙利铂联合用药在制备治疗胃癌药物中的应用 (Application of raloxifene and oxaliplatin combined medicine in preparation of medicine for treating gastric cancer ) 是由 李斌 于 2019-08-24 设计创作，主要内容包括：本发明属于医药领域,特别涉及雷洛昔芬与奥沙利铂联合用药在制备治疗胃癌药物中的应用。本发明首次提出雷洛昔芬与奥沙利铂联合用药在制备治疗胃癌药物中的应用,两者具有明显的协同作用,有效提高疗效,相对于单一组分疗效更为显著,提高了对肿瘤细胞的杀伤性；有效降低用药量,从而减少毒副作用。两者联合使用也可节约成本,减轻病人的经济负担,为胃癌的防治提供了一条新途径,在医药学领域有广阔的应用前景。(The invention belongs to the field of medicines, and particularly relates to application of raloxifene and oxaliplatin combined medicine in preparation of a medicine for treating gastric cancer. The invention provides the application of the raloxifene and oxaliplatin combined drug in preparing the drug for treating gastric cancer for the first time, the raloxifene and oxaliplatin have obvious synergistic effect, the curative effect is effectively improved, the curative effect is more obvious compared with that of a single component, and the killing property to tumor cells is improved; effectively reduces the dosage, thereby reducing the toxic and side effects. The combined use of the two can also save the cost, reduce the economic burden of patients, provide a new way for preventing and treating gastric cancer, and have wide application prospect in the field of medicine and pharmacology.)

1. Application of raloxifene and oxaliplatin combined medicine in preparation of medicine for treating gastric cancer.

2. Use according to claim 1, characterized in that: the dosage ratio of raloxifene to oxaliplatin is 0.5-128 ng/ml: 8 μ g/ml.

3. Use according to claim 1, characterized in that: the dosage ratio of raloxifene to oxaliplatin is 0.5-128 ng/ml: 16. mu.g/ml.

4. Use according to claim 1, characterized in that: the dosage ratio of raloxifene to oxaliplatin is 0.5-128 ng/ml: 32. mu.g/ml.

5. Use according to claim 1, characterized in that: the dosage ratio of raloxifene to oxaliplatin is 32 ng/ml: 16. mu.g/ml.

6. A preparation for treating gastric cancer is characterized in that: the composition consists of the composition as claimed in claims 1 to 5 and pharmaceutically acceptable auxiliary materials.

7. The formulation of claim 6, wherein: the preparation is in the form of injection, tablet, granule or capsule.

Technical Field

The invention belongs to the field of medicines, and particularly relates to application of raloxifene and oxaliplatin combined medicine in preparation of a medicine for treating gastric cancer.

Background

Gastric cancer is a common malignant tumor of the digestive tract, ranks third in the cancer-related deaths worldwide, and is one of the main health burdens of society. In China, gastric cancer is the second most prevalent cancer and the second leading cause of cancer death, so the development of anti-gastric cancer drugs is the hot spot of anti-tumor research and development in China. At present, the main treatment means of the gastric cancer is chemotherapy, but the single chemotherapy curative effect of the gastric cancer is poor, and the median survival time is only 8-13 months.

Oxaliplatin is a third generation platinum anticancer drug following cisplatin and carboplatin, has the characteristics of good anticancer effect, wide adaptation diseases and the like, shows broad-spectrum in-vitro cytotoxicity and in-vivo antitumor activity in various tumor model systems including human gastric cancer models, and plays an important role in the clinical treatment of gastric cancer. However, most of oxaliplatin is prone to cause severe gastrointestinal reactions, nephrotoxicity, ototoxicity, neurotoxicity and bone marrow toxicity during clinical use. Meanwhile, oxaliplatin is easy to cause acquired drug resistance, so that the actual curative effect of oxaliplatin is seriously reduced, and the clinical use of the oxaliplatin is further limited.

Raloxifene is a selective estrogen receptor modulator, has selective agonistic or antagonistic activity on tissues acted by estrogen, is mainly used for preventing and treating osteoporosis of postmenopausal women, and can remarkably reduce the incidence rate of vertebral body fracture. Reports on the effect of raloxifene in combination with oxaliplatin have not yet been made. The invention researches the effect of raloxifene in sensitizing oxaliplatin for treating gastric cancer in a human gastric cancer model.

Disclosure of Invention

Object of the Invention

The application of raloxifene sensitization oxaliplatin in the preparation of the medicine for treating gastric cancer is provided, the curative effect of the compound is enhanced, and a basis is provided for the new application of old medicines.

Technical scheme

The invention provides application of raloxifene and oxaliplatin combined medicine in preparation of a medicine for treating gastric cancer.

Therefore, the application of the combination of raloxifene and oxaliplatin in preparing the anti-gastric cancer medicament and the anti-gastric cancer medicament containing both raloxifene and oxaliplatin are all within the protection scope of the invention.

Preferably, the gastric cancer is human gastric cancer cell AGS or MGC 803.

Preferably, the gastric cancer is a human gastric cancer tissue organoid.

In addition, preferably, the dosage ratio of the raloxifene to the oxaliplatin is 0.5-128 ng/ml: 8 μ g/ml.

Preferably, the dosage ratio of raloxifene to oxaliplatin is 0.5-128 ng/ml: 16. mu.g/ml.

Preferably, the dosage ratio of raloxifene to oxaliplatin is 0.5-128 ng/ml: 32. mu.g/ml.

Particularly preferably, the dosage ratio of raloxifene to oxaliplatin is 32 ng/ml: 16. mu.g/ml.

Advantageous effects

The invention discloses the discovery of raloxifene on the aspect of improving the sensitivity of gastric cancer cells to oxaliplatin for the first time, can realize the synergistic effect, realize the obvious improvement of the anti-tumor effect, and reduce the dosage of oxaliplatin through combined medication so as to reduce the toxic and side effects, thereby having obvious significance for the application of raloxifene and oxaliplatin combined medication to the anti-gastric cancer.

Drawings

FIG. 1 is a graph showing the results of the inhibition analysis of Raloxifene alone in gastric cancer cells by MTT assay.

FIG. 2 is a graph showing the results of MTT method for detecting the inhibition rate of oxaliplatin alone or in combination with raloxifene and oxaliplatin for gastric cancer cell treatment; wherein, A: analyzing the result of cell AGS; b: cell MGC803 results analysis.

FIG. 3 is a graph showing the results of the inhibition rate analysis of oxaliplatin alone or in combination with raloxifene and oxaliplatin for human gastric cancer tissue organoids.

Detailed Description

The invention is further described with reference to the drawings and the following detailed description, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.

The kit materials used in the following examples are all commercially available unless otherwise specified.

Example 1 MTT assay to determine the sensitivity of different gastric cancer cells to Raloxifene

1. Experimental Material

(1) Medicine preparation: raloxifene hydrochloride is commonly used clinically, and the Raloxifene hydrochloride is used as a test medicament in the embodiment.

The chemical structural formula of the raloxifene hydrochloride is shown as follows:

(2) gastric cancer cell: human gastric cancer cells AGS and MGC 803.

(3) Commercial MTT kit.

2. Experiment grouping

(1) Control group: blank control, i.e. gastric cancer cells, were not treated with any drug.

(2) Experimental groups: gastric cancer cells were treated with different concentrations of raloxifene hydrochloride.

3. MTT method for detecting sensitivity of different tumor cells to raloxifene

(1) Human gastric cancer cells AGS and MGC803 are inoculated to a 96-well plate according to the number of 5000-9000 cells per well, and after the cells adhere to the wall, PBS solutions of raloxifene hydrochloride with different concentrations are respectively added. After 24 hours of incubation, 10. mu.l of 5mg/ml MTT was added to each well for another 4 hours, and then 100. mu.l of DMSO was added to each well after discarding the culture solution, and the absorbance of each well was read at 490nm using a microplate reader. Statistical analysis was performed on the data.

(2) The analysis result is shown in fig. 1, which indicates that raloxifene hydrochloride has a certain effect of inhibiting proliferation of gastric cancer cells, but the inhibition effect is weak.

Example 2 Effect of oxaliplatin alone or in combination with Raloxifene and oxaliplatin on gastric cancer cell inhibition Rate

1. Experimental Material

(1) Medicine preparation: raloxifene hydrochloride and oxaliplatin.

(2) Gastric cancer cell: human gastric cancer cells AGS and MGC 803.

(3) Commercial MTT kit.

2. Experiment grouping

(1) Control group: gastric cancer cells were treated with oxaliplatin only.

(2) Experimental groups: various concentrations of raloxifene hydrochloride and oxaliplatin are used in combination to treat gastric cancer cells.

3. MTT method for detecting sensitivity of gastric cancer cells to raloxifene and oxaliplatin combined drug

(1) Human gastric cancer cells AGS and MGC803 are inoculated to a 96-well plate according to the number of 5000-9000 cells per hole, and PBS solutions with different proportions of raloxifene hydrochloride and oxaliplatin are respectively added after the cells adhere to the wall. After 24 hours of incubation, 10. mu.l of 5mg/ml MTT was added to each well for another 4 hours, and then 100. mu.l of DMSO was added to each well after discarding the culture solution, and the absorbance of each well was read at 490nm using a microplate reader. Statistical analysis was performed on the data.

(2) The analysis result is shown in fig. 2, the addition of raloxifene on the basis of oxaliplatin can more obviously inhibit the proliferation of gastric cancer cells compared with the single drug, and the combined use of raloxifene and oxaliplatin shows that the treatment effect of gastric cancer is obviously improved and the synergistic treatment effect is realized.

Example 3 Effect of oxaliplatin alone or in combination with Raloxifene and oxaliplatin on human gastric carcinoma tissue organoid inhibition

1. Experimental Material

(1) Medicine preparation: raloxifene hydrochloride and oxaliplatin.

(2) Gastric cancer cell: human gastric cancer tissue organoids.

(3) The commercially available CellTiter-Glo 3D kit.

2. Experiment grouping

(1) Control group: treatment with oxaliplatin alone.

(2) Experimental groups: raloxifene hydrochloride and oxaliplatin at different concentrations are used in combination to treat human gastric cancer tissue organoids.

3. Sensitivity of human gastric cancer tissue organoids to combined administration of raloxifene and oxaliplatin

(1) Mixing the treated human gastric cancer cells with Matrigel, adding 50 mul of the mixed Mtrigel Matrigel into each well of a 48-well plate, adding 250 mul of complete culture medium into each well after gelling, and culturing in a cell culture box at 37 ℃ and with the concentration of CO2 being 5%. The culture solution is replaced every 3-4 days, and can be subcultured and frozen every 1-2 weeks. After the organoid modeling is successful, subculturing is carried out for drug treatment, culture solution containing different proportions of raloxifene hydrochloride and oxaliplatin is added into each hole, and the mixture is placed in an incubator for 5 days. Adding CellTiter-Glo 3D detection reagent with the same volume as the culture medium into each hole, shaking and uniformly mixing, incubating at room temperature for 25 minutes, detecting by using an enzyme-linked immunosorbent assay (ELISA) instrument, and performing statistical analysis on data.

(2) The analysis result is shown in figure 2, the addition of raloxifene on the basis of oxaliplatin can more obviously inhibit the cell proliferation of human gastric cancer tissue organoids compared with single drug, and the combined use of raloxifene and oxaliplatin shows that the treatment effect of gastric cancer is obviously improved and the synergistic treatment effect is realized.