Plant type medicine for inhibiting growth of tumor cells as well as preparation method and application of plant type medicine

Abstract

The invention provides a plant type medicine for inhibiting tumor cell growth as well as a preparation method and application thereof, and belongs to the technical field of medicines. According to the preparation method of the plant-type medicine for inhibiting the growth of the tumor cells, pretreatment steps such as low-temperature freezing treatment, vacuum drying treatment and ultrasonic disruption treatment are introduced, a proper amount of nitrogen is introduced into a reaction kettle, the oxidation-reduction environment is adjusted, and a degradation reconstruction chemical reaction is carried out in a microwave heating mode. The measures greatly improve the purity and the generation efficiency of the product from different aspects.

Description

Technical Field

[0001] The present application relates to the field of pharmaceutical technology. Specifically, it relates to a plant-based drug for inhibiting the growth of tumor cells, its preparation method, and its application. Background Art

[0002] Digestive tract cancers such as lung cancer, gastric cancer, colorectal cancer, liver cancer, and esophageal cancer are particularly common, which may be related to culture, dietary patterns, low intake of food nutrients, and food contamination.

[0003] Currently, for the treatment of cancer, chemotherapy is one of the most common means. However, anti-tumor chemotherapy drugs have problems such as high costs and drug resistance, and the 5-year survival rate of tumor patients treated with chemotherapy is still less than 25%. Such data is deeply worrying and also prompts the medical community to continuously explore new treatment methods.

[0004] Against this background, the plant-based drug for inhibiting the growth of tumor cells of the present invention came into being. This drug is made from natural plants through scientific extraction and refinement. Compared with traditional chemotherapy drugs, it has several significant advantages. First, it uses natural plants as raw materials, which are widely sourced and have low costs, and is expected to reduce the treatment costs of patients. Second, while inhibiting the growth of tumor cells, this drug has less toxic effects on normal cells, reducing the occurrence of side effects. In addition, in response to the problem that chemotherapy drugs are prone to drug resistance, the drug of the present invention reduces the generation of drug resistance and improves the treatment effect through a unique mechanism of action. For the existing extraction methods, Chinese invention patent, a plant-based drug for inhibiting sexually transmitted viruses HIV-1 and HSV-2 and its preparation method, CN115252693A, has low extraction purity and low reaction efficiency. Summary of the Invention

[0005] The purpose of the present application is to provide a plant-based drug for inhibiting the growth of tumor cells, its preparation method, and its application, which can improve the low extraction purity and enhance the reaction efficiency.

[0006] In the first aspect, the present application provides a preparation method of a plant-based drug for inhibiting the growth of tumor cells, including the following steps Perform cryogenic freezing treatment on the biomass raw material, and after natural thawing after cryogenic freezing treatment, perform vacuum drying treatment. After repeating this step 3 to 5 times, form a pre-treated biomass raw material; Use the high-frequency vibration of ultrasonic waves to crush the pre-treated biomass raw material; Regulate the reaction kettle, fill it with an appropriate amount of nitrogen, adjust the redox environment, add the crushed biomass raw material to the reaction kettle, and mix it with tartaric acid, ferrous sulfate, hydrogen peroxide aqueous solution, and deionized water and stir. Perform a degradation and reconstruction chemical reaction under the action of cellulase catalysis and microwave heating; After the reaction is completed, the generated steam is guided to a condenser for cooling to obtain a plant degradation and reconstruction extract.

[0007] In a possible embodiment, the low-temperature freezing temperature is -15°C to -25°C.

[0008] In one possible embodiment, the cryogenic freezing temperature is -20°C.

[0009] In a possible implementation manner, the microwave power of microwave heating is 500-1000 W and the frequency is 2450 MHz.

[0010] In one possible implementation, the stirring speed is 200-400 rpm.

[0011] In a possible embodiment, the biomass raw material is Salix psammophila or Caragana korshinskii.

[0012] In a second aspect, the present application provides a plant-based drug for inhibiting tumor cell growth, which is prepared using the aforementioned method for preparing a plant-based drug for inhibiting tumor cell growth.

[0013] In a third aspect, the present application provides a plant-based drug for inhibiting tumor cell growth for use in a drug for inhibiting tumor cell growth.

[0014] The present application provides a plant-based drug for inhibiting tumor cell growth, a preparation method thereof, and the beneficial effects of the application include: the plant-based drug for inhibiting tumor cell growth, first, by introducing pretreatment steps such as low-temperature freezing treatment, vacuum drying treatment, and ultrasonic crushing treatment, the technical solution of the present invention can more effectively crush the biomass raw material, increase its specific surface area and reaction activity, thereby improving the efficiency and purity of the subsequent degradation and reconstruction chemical reaction. These pretreatment steps can destroy the cell wall structure of the biomass raw material, making it easier to release the active ingredients inside. Secondly, the technical solution of the present invention is filled with an appropriate amount of nitrogen in the reactor to adjust the redox environment, which helps to control the process and product of the degradation and reconstruction chemical reaction. Nitrogen, as an inert gas, can reduce the oxidation reaction during the reaction process and avoid the generation of unnecessary by-products. In addition, the present invention also uses microwave heating to perform degradation and reconstruction chemical reactions. Microwave heating is fast, uniform, and efficient, which can enable the reaction system to reach the desired reaction temperature in a short time, and can generate more collision opportunities between the reactant molecules, thereby accelerating the reaction rate. These measures greatly improve the purity and generation efficiency of the product from different aspects.

[0015] In summary, the technical solution of the present invention significantly improves the preparation efficiency and purity of plant-based drugs for inhibiting the growth of tumor cells by introducing new pretreatment steps, adjusting the reaction environment, and using microwave heating. These improvements not only help reduce the treatment cost for patients and improve the treatment effect, but also provide new ideas and methods for the research and development and application of plant-based drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present application, and thus should not be regarded as limiting the scope. For those of ordinary skill in the art, other related drawings can also be obtained based on these drawings without creative efforts, which also fall within the protection scope of the present application.

[0017] Figure 1 Effect diagram of the inhibitory effect of plant degradation and reconstruction extracts with different concentrations provided by the embodiments of the present application on the growth of Eca cells (human lung cancer cells).

[0018] Figure 2 Effect diagram of the inhibitory effect of plant degradation and reconstruction extracts with different concentrations provided by the embodiments of the present application on the growth of HepG2 cells (human liver cancer cells).

[0019] Figure 3 Effect diagram of the inhibitory effect of plant degradation and reconstruction extracts with different concentrations provided by the embodiments of the present application on the growth of HCT116 cells (human colon cancer cells).

[0020] Figure 4 Effect diagram of the inhibitory effect of plant degradation and reconstruction extracts with different concentrations provided by the embodiments of the present application on the growth of MCF cells (human breast cancer cells).

[0021] Figure 5 Effect diagram of the inhibitory effect of plant degradation and reconstruction extracts with different concentrations provided by the embodiments of the present application on the growth of Hela cells (human cervical cancer cells). DETAILED DESCRIPTION OF THE EMBODIMENTS

[0022] The present application provides a method for preparing a plant-based drug for inhibiting the growth of tumor cells, including the following steps: The biomass raw material is subjected to cryogenic freezing treatment, naturally thawed after cryogenic freezing treatment, and then vacuum dried. This step is cycled 3 to 5 times to form a pretreated biomass raw material.

[0023] The pretreated biomass raw material is crushed by using the high-frequency vibration effect of ultrasonic waves.

[0024] Regulate the reactor, fill in an appropriate amount of nitrogen, adjust the redox environment, add the biomass raw material after crushing treatment to the reactor, and mix and stir it with tartaric acid, ferrous sulfate, hydrogen peroxide aqueous solution, and deionized water. Carry out a degradation and reconstruction chemical reaction under the action of cellulase catalysis and microwave heating.

[0025] After the reaction ends, guide the generated steam to a condenser for cooling to obtain a plant degradation and reconstruction extract.

[0026] Example A method for preparing a plant-based drug for inhibiting the growth of tumor cells includes the following steps. Perform cryogenic freezing treatment on the biomass raw material of Salix psammophila or Caragana korshinskii. The cryogenic freezing temperature is -15°C. After natural thawing after cryogenic freezing treatment, perform vacuum drying treatment. Repeat this step 3 times to form a pretreated biomass raw material.

[0027] Use the high-frequency vibration of ultrasonic waves to crush the pretreated biomass raw material.

[0028] Regulate the reactor, fill in an appropriate amount of nitrogen, adjust the redox environment, add the crushed biomass raw material to the reactor, and mix and stir it with tartaric acid, ferrous sulfate, hydrogen peroxide aqueous solution, and deionized water. The stirring speed is 200 rpm. Carry out a degradation and reconstruction chemical reaction under the action of cellulase catalysis and microwave heating. Among them, the microwave power of microwave heating is 500 w, and the frequency is 2450 MHz.

[0029] After the reaction ends, guide the generated steam to a condenser for cooling to obtain a plant degradation and reconstruction extract.

[0030] Example A method for preparing a plant-based drug for inhibiting the growth of tumor cells includes the following steps. Perform cryogenic freezing treatment on the biomass raw material of Salix psammophila or Caragana korshinskii. The cryogenic freezing temperature is -20°C. After natural thawing after cryogenic freezing treatment, perform vacuum drying treatment. Repeat this step 4 times to form a pretreated biomass raw material.

[0031] Use the high-frequency vibration of ultrasonic waves to crush the pretreated biomass raw material.

[0032] Regulate the reactor, fill in an appropriate amount of nitrogen, adjust the redox environment, add the crushed biomass raw material to the reactor, and mix and stir it with tartaric acid, ferrous sulfate, hydrogen peroxide aqueous solution, and deionized water. The stirring speed is 300 rpm. Carry out a degradation and reconstruction chemical reaction under the action of cellulase catalysis and microwave heating. Among them, the microwave power of microwave heating is 750 w, and the frequency is 2450 MHz.

[0033] After the reaction is completed, the generated steam is guided to a condenser for cooling to obtain a plant degradation and reconstruction extract.

[0034] Example A method for preparing a plant-based drug for inhibiting the growth of tumor cells, comprising the following steps: The biomass raw materials of Salix psammophila or Caragana korshinskii are subjected to cryogenic freezing treatment at a cryogenic freezing temperature of -25°C.

[0035] After natural thawing after cryogenic freezing treatment, vacuum drying treatment is carried out. After this step is cycled 5 times, a pre-treated biomass raw material is formed.

[0036] The pre-treated biomass raw material is crushed by using the high-frequency vibration of ultrasonic waves.

[0037] The reaction kettle is regulated, an appropriate amount of nitrogen is filled, the redox environment is adjusted, the crushed biomass raw material is added to the reaction kettle, and it is mixed and stirred with tartaric acid, ferrous sulfate, hydrogen peroxide aqueous solution, and deionized water. The stirring speed is 400 rpm, and a degradation and reconstruction chemical reaction is carried out under the action of cellulase catalysis and microwave heating. Among them, the microwave power of microwave heating is 1000 w, and the frequency is 2450 MHz.

[0038] After the reaction is completed, the generated steam is guided to a condenser for cooling to obtain a plant degradation and reconstruction extract.

[0039] This application provides a method for preparing a plant-based drug for inhibiting the growth of tumor cells. In this method for preparing a plant-based drug for inhibiting the growth of tumor cells, first, by introducing pre-treatment steps such as cryogenic freezing treatment, vacuum drying treatment, and ultrasonic crushing treatment, the technical solution of the present invention can more effectively crush the biomass raw material, increase its specific surface area and reaction activity, thereby improving the efficiency and purity of the subsequent degradation and reconstruction chemical reaction. These pre-treatment steps can destroy the cell wall structure of the biomass raw material, making the active ingredients inside it easier to be released. Secondly, the technical solution of the present invention fills an appropriate amount of nitrogen in the reaction kettle to adjust the redox environment, which helps to control the process and products of the degradation and reconstruction chemical reaction. As an inert gas, nitrogen can reduce the oxidation reaction during the reaction and avoid the generation of unnecessary by-products. In addition, the present invention also uses microwave heating for the degradation and reconstruction chemical reaction. Microwave heating has the characteristics of fast speed, uniformity, and high efficiency. It can make the reaction system reach the required reaction temperature in a short time, and can make the reactant molecules have more collision opportunities, thereby accelerating the reaction rate. These measures greatly improve the purity and production efficiency of the product from different aspects.

[0040] The present invention provides a plant-based drug for inhibiting the growth of tumor cells, which is prepared by using the aforementioned method for preparing a plant-based drug for inhibiting the growth of tumor cells.

[0041] The present invention provides an application of a method for preparing a plant-based drug for inhibiting the growth of tumor cells in a drug for inhibiting the growth of tumor cells.

[0042] Here, the tumor cells can be tumor cells such as Eca (lung cancer) cells, HepG2 (liver cancer) cells, HCT116 (colon cancer) cells, and Hela (cervical cancer) cells, or can also be tumor cells such as leukemia, glioma, pancreatic cancer, lymphoma, esophageal cancer, rectal cancer, and gastric cancer.

[0043] Taking Example 2 as an example below, the effect of the plant-based drug for inhibiting the growth of tumor cells of the present invention in inhibiting the growth of tumor cells is described.

[0044] In order to study the inhibitory effect of plant degradation and reconstruction extracts at different concentrations on the growth of tumor cells of lung cancer, liver cancer, colon cancer, and cervical cancer, the control group uses fluorouracil (5-Fu), and is compared with fluorouracil (5-Fu). The following test methods can be carried out: Test preparation Prepare cell lines: Obtain tumor cell lines of lung cancer, liver cancer, colon cancer, and cervical cancer, ensure the reliable source, high purity, and good proliferation activity of the cell lines.

[0045] Prepare drugs: Prepare plant degradation and reconstruction extracts at different concentrations, and prepare an appropriate amount of fluorouracil (5-Fu) as the control group drug.

[0046] Prepare cell culture supplies: including cell culture medium, serum, antibiotics, cell culture plates, cell culture flasks, pipettes, pipettes, etc.

[0047] Set up experimental groups and control groups: Set up experimental groups and control groups for each type of tumor cell. The experimental groups are treated with plant degradation and reconstruction extracts at different concentrations, and the control groups are treated with fluorouracil (5-Fu).

[0048] Cell culture and treatment Cell seeding: Seed the tumor cells into cell culture plates or cell culture flasks at an appropriate density, and add an appropriate amount of cell culture medium to ensure the normal growth of the cells.

[0049] Drug treatment: After the cells adhere and grow, add plant degradation and reconstruction extracts at different concentrations to the experimental groups, and add fluorouracil (5-Fu) to the control groups, and treat for a certain period of time (such as 24 hours, 48 hours, 72 hours, etc.).

[0050] Observe the cell growth situation: During the drug treatment period, regularly observe the cell growth situation, and record phenomena such as changes in cell morphology, slowdown of proliferation rate, or death.

[0051] Cell viability detection Use appropriate cell viability detection methods (such as MTT method, CCK-8 method, etc.) to detect the viability of tumor cells in the experimental group and the control group.

[0052] According to the detection results, draw a cell growth curve or a cell survival rate curve to compare the inhibitory effects of different concentrations of the plant degradation and reconstruction extract and 5-fluorouracil (5-Fu) on the growth of tumor cells.

[0053] Data analysis and conclusion Statistically analyze the experimental data to compare the significance of differences between the experimental group and the control group.

[0054] Based on the data analysis results, obtain the inhibitory effects of different concentrations of the plant degradation and reconstruction extract on the growth of tumor cells in lung cancer, liver cancer, colon cancer, and cervical cancer, and compare them with the inhibitory effect of 5-fluorouracil (5-Fu).

[0055] Combined with the cell viability detection results and the cell growth observation, comprehensively evaluate the inhibitory effect of the plant degradation and reconstruction extract on tumor cells.

[0056] Through the above experimental methods, the inhibitory effects of different concentrations of the plant degradation and reconstruction extract on the growth of various tumor cells can be systematically studied and compared with the commonly used clinical chemotherapy drug 5-fluorouracil (5-Fu), providing a scientific basis for the application of the plant degradation and reconstruction extract in the anti-tumor field.

[0057] Please refer to Figure 1 , the effect diagram of the inhibitory effect of different concentrations of the plant degradation and reconstruction extract on the growth of Eca cells (human lung cancer cells). It shows that the inhibitory effect of the reconstruction extract on Eca (human lung cancer cells) is stronger at dilutions of 60 - 500 times, and the inhibitory effect on cells is not significant at dilutions above 500 times, and the inhibitory effect is stronger than that of 5-fluorouracil (5-Fu) (5-fluorouracil is a commonly used clinical chemotherapy drug for malignant tumors).

[0058] Please refer to Figure 2 , the effect diagram of the inhibitory effect of different concentrations of the plant degradation and reconstruction extract on the growth of HepG2 cells (human liver cancer cells). It shows that the inhibitory effect of the reconstruction extract on HepG2 cells (human liver cancer cells) is stronger at dilutions of 100 times, 125 times, and 250 times, and the inhibitory effect on cells is not significant at dilutions above 500 times.

[0059] Please refer to Figure 3, Effect diagram of the growth inhibitory effect of plant degradation and reconstruction extracts at different concentrations on HCT116 cells (human colon cancer cells). It shows that the inhibitory effect of the reconstruction extract on HCT116 cells (human colon cancer cells) is stronger at dilutions of 100 times and 125 times, and the inhibitory effect on cells is not significant when diluted more than 250 times.

[0060] Please refer to Figure 4 , Effect diagram of the growth inhibitory effect of plant degradation and reconstruction extracts at different concentrations on Hela cells (human cervical cancer cells). It shows that the inhibitory effect of the reconstruction extract on Hela cells (human cervical cancer cells) is stronger at dilutions of 60 - 500 times, the inhibitory effect on cells is not significant when diluted more than 500 times, and the inhibitory effect is stronger than that of 5-fluorouracil (5-Fu).

[0061] In summary, the experimental conclusions on the growth inhibitory effect of the plant degradation and reconstruction extract of the present invention on tumor cells such as lung cancer, liver cancer, colon cancer, and cervical cancer: The plant degradation and reconstruction extract has obvious anti-tumor activity on Eca (lung cancer) cells, HepG2 (liver cancer) cells, HCT116 (colon cancer) cells, and Hela (cervical cancer) cells at dilutions of 60 - 500 times, and the anti-tumor effect is not obvious when the dilution multiple is greater than 500 times. The anti-tumor activity on Eca (lung cancer) cells and Hela (cervical cancer) cells is stronger than that of 5-fluorouracil.

[0062] For the preparation method of the plant-based drug for inhibiting tumor cell growth provided in this application, taking Example 2 as an example, compared with CN115252693A (hereinafter referred to as the prior art), its extraction purity and production efficiency are as follows in the table: In addition, for the preparation method of the plant-based drug for inhibiting tumor cell growth provided in this application, taking Example 2 as an example, compared with the prior art CN115252693A, its purity is increased from 85.07% to 98.75%, and the production efficiency is increased by 58%.

[0063] The embodiments described above are some embodiments of this application, not all embodiments. The detailed description of the embodiments of this application is not intended to limit the scope of this application claimed, but merely represents the selected embodiments of this application. All other embodiments obtained by those of ordinary skill in the art based on the embodiments in this application without creative efforts belong to the scope of protection of this application.

Claims

1. A preparation method of a plant-based drug for inhibiting the growth of tumor cells, characterized in that, It includes the following steps: The biomass raw material is subjected to low-temperature freezing treatment, naturally thawed after the low-temperature freezing treatment, and then vacuum dried. This step is cycled 3 to 5 times to form the pretreated biomass raw material; The pretreated biomass raw material is crushed by the high-frequency vibration of ultrasonic waves; The reaction kettle is regulated, an appropriate amount of nitrogen is charged, the redox environment is adjusted, the crushed biomass raw material is added to the reaction kettle, and it is mixed and stirred with tartaric acid, ferrous sulfate, hydrogen peroxide aqueous solution, and deionized water, and a degradation and reconstruction chemical reaction is carried out under the action of cellulase catalysis and microwave heating; After the reaction is completed, the generated steam is guided to a condenser for cooling to obtain a plant degradation and reconstruction extract.

2. The preparation method of the plant-based drug for inhibiting the growth of tumor cells according to claim 1, characterized in that, The temperature of the low-temperature freezing is -15°C to -25°C.

3. The method for preparing a plant-based drug for inhibiting the growth of tumor cells according to claim 2, wherein The temperature of the low-temperature freezing is -20°C.

4. The method for preparing a plant-based drug for inhibiting the growth of tumor cells according to claim 1, wherein The microwave power of the microwave heating is 500 to 1000 W, and the frequency is 2450 MHz.

5. The method for preparing a plant-based drug for inhibiting the growth of tumor cells according to claim 1, wherein The stirring speed is 200 to 400 rpm.

6. The preparation method of the plant-based drug for inhibiting the growth of tumor cells according to claim 1, characterized in that, The biomass raw material is Salix psammophila or Caragana korshinskii.

7. A plant-based drug for inhibiting the growth of tumor cells, characterized in that, It is prepared by using the preparation method of the plant-type drug for inhibiting the growth of tumor cells according to any one of claims 1-6.

8. An application of the preparation method of the plant-type drug for inhibiting the growth of tumor cells according to any one of claims 1-6 in the drug for inhibiting the growth of tumor cells.

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