Anti-tumor composition as well as preparation method and application thereof

Abstract

The invention provides an anti-tumor composition as well as a preparation method and application thereof, and relates to the technical field of biological medicines. The anti-tumor composition disclosed by the invention is prepared from the following raw materials in parts by mass: 18 to 22 parts of ginsenoside Rg3, 3 to 7 parts of ginsenoside CK, 28 to 32 parts of ganoderma lucidum polysaccharide, 23 to 27 parts of astragalus polysaccharide, 8 to 12 parts of lentinan, 16 to 20 parts of panax notoginseng saponins, 9 to 11 parts of resveratrol, 13 to 17 parts of curcumin, 6 to 10 parts of dihydromyricetin and 7 to 9 parts of sophocarpidine. The ratio of the total mass of the ganoderan and the astragalus polysaccharide to the total mass of the ginsenoside Rg3 and the panax notoginseng saponins is (1.3-1.6): 1. Through a multi-target and multi-path synergistic effect mechanism, tumor cell proliferation can be directly inhibited, apoptosis can be induced, the cell cycle can be retarded, tumor angiogenesis can be inhibited, the immune function of the body can be enhanced, the antioxidant effect can be exerted, tumor invasion and metastasis can be inhibited, meanwhile, the obvious synergistic and toxicity reducing effects can be shown in combined chemotherapy application, and the application prospect is wide. And a new technical scheme is provided for comprehensive treatment of tumors.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine, in particular to an anti-tumor composition and a preparation method and application thereof. BACKGROUND

[0002] Malignant tumor has become a major global public health problem, and its incidence and mortality continue to rise. Traditional treatment methods such as surgery, radiotherapy and chemotherapy are widely used in clinical practice, but they generally have poor selectivity, serious side effects, and are prone to drug resistance, which seriously affects the quality of life and treatment compliance of patients. Chemotherapy drugs represented by cisplatin often cause bone marrow suppression, liver and kidney function damage and other serious adverse reactions while killing tumor cells, forcing many patients to discontinue treatment or reduce the dose, thereby affecting the efficacy.

[0003] Traditional Chinese medicine has accumulated rich clinical experience in the field of tumor prevention and treatment, and has the characteristics of multi-target and multi-pathway action, which can not only directly inhibit tumor growth, but also regulate immune function and reduce the side effects of radiotherapy and chemotherapy. However, current research on traditional Chinese medicine for anti-tumor is mostly focused on the mechanism of single effective component, which often needs a high dose to achieve the desired effect, and the target is relatively single, making it difficult to fully exert the anti-tumor effect. Although some studies have tried to use multiple components of traditional Chinese medicine in combination, most of them lack systematic compatibility theory guidance and dose-effect relationship research, and the synergistic mechanism between components is not clear, resulting in insufficient scientificity and stability of the prescription.

[0004] In particular, in the application of compatibility of polysaccharide components and saponin components, there is still a lack of in-depth research on the optimal ratio and synergistic effect of the two components. Polysaccharide components mainly play an indirect anti-tumor effect by enhancing immune function, while saponin components have a direct inhibitory effect on tumor cell proliferation. How to scientifically match the two components to achieve complementary advantages and synergistic effect is still a key scientific problem to be solved.

[0005] In addition, the existing traditional Chinese medicine composition has limited effect on reducing the side effects of chemotherapy drugs when used in combination with chemotherapy, and cannot meet the dual needs of "synergistic effect and reduction of toxicity" in clinical practice. Therefore, it is of great clinical application value and practical significance to develop a traditional Chinese medicine compound preparation that has significant anti-tumor activity and can reduce the side effects of chemotherapy drugs. SUMMARY

[0006] In view of this, the present invention provides an antitumor composition, its preparation method, and its application. Addressing the technical problems in existing technologies such as the limited antitumor effects of single effective components of traditional Chinese medicine, the lack of scientific basis for the compatibility of multiple component combinations, and the unclear synergistic mechanism, the present invention utilizes a multi-target, multi-pathway synergistic mechanism to directly inhibit tumor cell proliferation, induce apoptosis, and arrest the cell cycle. It also inhibits tumor angiogenesis, enhances the body's immune function, exerts antioxidant effects, and inhibits tumor invasion and metastasis. Furthermore, it exhibits significant synergistic and toxicity-reducing effects when used in combination with chemotherapy, providing a new technical solution for the comprehensive treatment of tumors.

[0007] A first aspect of the present invention is to provide an antitumor composition comprising, by weight, the following raw materials: Ginsenoside Rg3 18-22 parts, ginsenoside CK 3-7 parts, Ganoderma lucidum polysaccharide 28-32 parts, Astragalus polysaccharide 23-27 parts, Lentinan 8-12 parts, Panax notoginseng total saponins 16-20 parts, resveratrol 9-11 parts, curcumin 13-17 parts, dihydromyricetin 6-10 parts, matrine 7-9 parts; The ratio of the total mass of Ganoderma lucidum polysaccharides and Astragalus polysaccharides to the total mass of ginsenoside Rg3 and Panax notoginseng total saponins is (1.3-1.6):1.

[0008] Preferably, the antitumor composition comprises, by weight, the following raw materials: Ginsenoside Rg3 20 parts, ginsenoside CK 5 parts, Ganoderma lucidum polysaccharide 30 parts, Astragalus polysaccharide 25 parts, Lentinus edodes polysaccharide 10 parts, Panax notoginseng total saponins 18 parts, resveratrol 10 parts, curcumin 15 parts, dihydromyricetin 8 parts, matrine 8 parts.

[0009] The raw materials of this invention can be commercially available products or prepared using conventional methods in the art.

[0010] Preferably, the antitumor composition can be prepared into different dosage forms by adding appropriate amounts of pharmaceutical excipients.

[0011] Preferably, the dosage form is a powder, capsule, tablet, or injection.

[0012] A second aspect of the present invention is to provide a method for preparing the antitumor composition, specifically comprising the following steps: Weigh the raw materials according to the proportions, mix them evenly, and obtain the antitumor composition.

[0013] A third aspect of the present invention is to provide an antitumor composition for use in the preparation of antitumor drugs or in combination with chemotherapy drugs to enhance efficacy and reduce toxicity.

[0014] While traditional Chinese medicine decoctions are a commonly used dosage form in clinical practice, they face numerous technical bottlenecks. The composition of these decoctions is complex, containing a large number of ineffective or low-efficacy components such as starch, cellulose, and pigments. These impurities not only dilute the concentration of active ingredients but may also cause gastrointestinal discomfort. Significant differences in the origin, harvesting period, and processing methods of different batches of medicinal materials lead to large fluctuations in the quality of the decoctions, making standardization and quality control difficult. More importantly, the extraction rate of each active ingredient in the decoction is affected by factors such as decoction time, temperature, and solvent ratio. The optimal extraction conditions for certain fat-soluble components, such as ginsenoside Rg3 and resveratrol, and water-soluble components, such as polysaccharides, are contradictory, making it impossible to achieve maximum extraction efficiency in the same decoction process. Furthermore, the poor stability of decoctions, their difficulty in storage and transportation, and the large dosages required lead to poor patient compliance. These inherent defects limit the clinical application and modernization of traditional Chinese medicine.

[0015] This application creatively employs a scientific formulation of single active ingredients with known pharmacological activities from traditional Chinese medicinal materials, achieving a significant technological breakthrough. By using standardized active ingredients with high purity, the dosage of each ingredient can be precisely controlled, establishing a clear dose-effect relationship. Experimental data shows that the optimal ratio of polysaccharides to saponins is (1.3-1.6):1, a precise ratio that cannot be achieved with traditional decoctions. Different components require different extraction methods; saponins require alcohol extraction, and polysaccharides require water extraction. Using purified ingredients ensures that each active substance reaches its highest content and optimal activity state, and then they are compounded in the optimal ratio to maximize synergistic effects.

[0016] The inventive essence of this application lies in realizing a paradigm shift from empirical formulation to rational design. Traditional decoctions rely on the "principal, assistant, adjuvant, and guide" theory of traditional Chinese medicine, but the material basis and mechanism of action among the various medicinal materials are unclear. Based on modern pharmacology, this application clarifies the anti-tumor targets of each of the 10 effective components, achieving a "1+1>2" effect through multi-target synergy.

[0017] This invention establishes an antitumor composition with ginsenoside Rg3, Ganoderma lucidum polysaccharide, and Panax notoginseng total saponins as the core, combined with ginsenoside CK, Astragalus polysaccharide, Lentinan, resveratrol, curcumin, dihydromyricetin and matrine, etc., through compatibility studies and dose-effect relationship optimization. The components produce significant synergistic effects.

[0018] First, this invention establishes the optimal ratio of polysaccharide components (Ganoderma lucidum polysaccharides and Astragalus polysaccharides) to saponin components (ginsenoside Rg3 and total Panax notoginseng saponins) as (1.3-1.6):1. This ratio achieves the best synergistic effect between immunomodulatory and direct antitumor activities. In vitro experiments show that the composition of this invention significantly inhibits the proliferation of human hepatocellular carcinoma HepG2 cells compared to the control group with a single component or an unbalanced ratio. Mouse xenograft experiments further confirm that the tumor inhibition rate of the composition of this invention is significantly higher than that of the control group with improper ratios or missing components.

[0019] Secondly, the composition of this invention has a significant immune-enhancing effect. Animal experiments showed that the spleen index and thymus index of mice in the treatment group were significantly higher than those in the model control group, indicating that the composition can effectively enhance the body's immune function while inhibiting tumor growth, which is difficult to achieve with a single chemotherapy drug. This "combined attack and supplementation" effect makes this invention more in line with the concept of comprehensive tumor treatment.

[0020] Finally, this invention demonstrates excellent synergistic and toxicity-reducing effects in combination chemotherapy. When used in combination with cisplatin, the composition of this invention not only enhances tumor inhibition rates but, more importantly, significantly reduces the toxic side effects of chemotherapy drugs. The white blood cell count in the combination therapy group was significantly higher than that in the cisplatin-only group, effectively alleviating bone marrow suppression. Simultaneously, liver function indicators ALT and kidney function indicators Cr were also significantly lower than those in the cisplatin-only group, indicating that this invention has a significant protective effect against chemotherapy-induced liver and kidney damage. This dual effect of synergistic efficacy and toxicity reduction provides new insights for optimizing clinical chemotherapy regimens.

[0021] Compared with the prior art, the beneficial technical effects of the present invention are as follows: The composition of this invention utilizes a variety of natural active ingredients, exhibiting high safety, no significant toxic side effects with long-term use, and stable and controllable quality. Through scientific formulation, each component exerts a synergistic effect across multiple targets and pathways in the anti-tumor process. It can directly inhibit tumor cell proliferation, induce apoptosis, and arrest the cell cycle, while also inhibiting tumor angiogenesis, regulating immune function, exerting antioxidant effects, and inhibiting tumor invasion and metastasis, thus forming a comprehensive anti-tumor effect with broad application prospects and significant clinical promotion value. Detailed Implementation

[0022] The technical solutions in the embodiments of this application will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of this application, not all of them. Other embodiments obtained by those skilled in the art based on the embodiments of this application without creative effort are all within the scope of protection of the invention. In the following embodiments, all raw materials are commercially available.

[0023] Unless otherwise specified, all experiments were repeated three times. Results are expressed as mean ± standard deviation, and P < 0.05 indicates a significant difference.

[0024] Example 1 An antitumor composition, comprising the following parts by weight: Ginsenoside Rg3 20 parts, ginsenoside CK 5 parts, Ganoderma lucidum polysaccharide 30 parts, Astragalus polysaccharide 25 parts, Lentinus edodes polysaccharide 10 parts, Panax notoginseng total saponins 18 parts, resveratrol 10 parts, curcumin 15 parts, dihydromyricetin 8 parts, matrine 8 parts. The method for preparing the antitumor composition is as follows: Accurately weigh each active ingredient according to the formula, pass them through a 100-mesh sieve, mix them thoroughly, and obtain the antitumor composition.

[0025] Example 2 An antitumor composition, comprising the following parts by weight: Ginsenoside Rg3 18 parts, ginsenoside CK 3 parts, Ganoderma lucidum polysaccharide 28 parts, Astragalus polysaccharide 23 parts, Lentinus edodes polysaccharide 8 parts, Panax notoginseng total saponins 16 parts, resveratrol 9 parts, curcumin 13 parts, dihydromyricetin 6 parts, matrine 7 parts. The preparation method is the same as in Example 1.

[0026] Example 3 An antitumor composition, in parts by weight, has the following composition: Ginsenoside Rg3 22 parts, ginsenoside CK 7 parts, Ganoderma lucidum polysaccharide 32 parts, Astragalus polysaccharide 27 parts, Lentinus edodes polysaccharide 12 parts, Panax notoginseng total saponins 20 parts, resveratrol 11 parts, curcumin 17 parts, dihydromyricetin 10 parts, matrine 9 parts. The preparation method is the same as in Example 1.

[0027] Example 4: Preparation of Capsules Take 100 g of the powdered composition prepared in Example 1, add 50 g of microcrystalline cellulose, 10 g of sodium carboxymethyl starch, and 2 g of magnesium stearate, and mix thoroughly. Fill the mixture into No. 0 capsule shells, with each capsule containing 0.4 g (0.25 g of the composition), to obtain 400 capsules.

[0028] Example 5: Preparation of Tablets Take 100 g of the powdered composition prepared in Example 1, add 80 g of microcrystalline cellulose, 15 g of sodium carboxymethyl starch, 10 g of povidone K30, and 3 g of magnesium stearate, and mix well after passing through an 80-mesh sieve. Perform wet granulation using 95% ethanol as a wetting agent to prepare a soft mass, granulate through a 20-mesh sieve, dry at 60°C, granulate, and then compress into tablets.

[0029] Comparative Example 1: An antitumor composition, in parts by weight, has the following composition: Ginsenoside Rg3 20 parts, Ganoderma lucidum polysaccharide 30 parts, Astragalus polysaccharide 25 parts, Lentinan shiitake polysaccharide 10 parts, Panax notoginseng total saponins 18 parts, Resveratrol 10 parts, Curcumin 15 parts, Dihydromyricetin 8 parts, Matrine 8 parts.

[0030] Comparative Example 2: An antitumor composition, in parts by weight, has the following composition: Ginsenoside Rg3 20 parts, ginsenoside CK 5 parts, Ganoderma lucidum polysaccharide 20 parts, Astragalus polysaccharide 18 parts, Lentinan shiitake polysaccharide 10 parts, Panax notoginseng total saponins 18 parts, resveratrol 10 parts, curcumin 15 parts, dihydromyricetin 8 parts, matrine 8 parts.

[0031] Comparative Example 3: An antitumor composition, in parts by weight, has the following composition: Ginsenoside Rg3 20 parts, ginsenoside CK 5 parts, Ganoderma lucidum polysaccharide 40 parts, Astragalus polysaccharide 35 parts, Lentinus edodes polysaccharide 10 parts, Panax notoginseng total saponins 18 parts, resveratrol 10 parts, curcumin 15 parts, dihydromyricetin 8 parts, matrine 8 parts.

[0032] Comparative Example 4: An antitumor composition, in parts by weight, has the following composition: Ginsenoside Rg3 20 parts, Panax notoginseng total saponins 18 parts.

[0033] Comparative Example 5: An antitumor composition, in parts by weight, has the following composition: The difference from Example 1 is that each component is in portions of 10.

[0034] Test Example 1: In vitro antitumor activity experiment Frozen HepG2 cells were revived and cultured in DMEM complete medium containing 10% FBS, 100 U / mL penicillin and 100 μg / mL streptomycin. The cells were placed in a 37°C, 5% CO2 incubator. When the cells adhered and grew to 80%-90% confluence, they were passaged by digestion with 0.25% trypsin, and cells in the logarithmic growth phase were selected for experiments.

[0035] First, cells in the logarithmic growth phase were collected, digested with trypsin, and prepared into a single-cell suspension. Cells were then counted using a hemocytometer, and the cell density was adjusted to 5 × 10⁶ cells / mL. 4 Cells / mL, add 100 μL of cell suspension (containing 5 × 10⁶ cells / mL) to each well of a 96-well plate. 3Cells were cultured for 24 hours to allow them to adhere to the culture medium. Then, drug treatment was performed: the culture medium was discarded, and the cells were washed once with PBS. The following groups were set up: a blank control group (100 μL of complete culture medium), a solvent control group (100 μL of culture medium containing 0.1% DMSO), an example group (100 μL of culture medium containing different example samples, with final concentrations of 25, 50, 100, and 200 μg / mL), a comparative group (100 μL of culture medium containing different comparative sample samples, with a final concentration of 100 μg / mL), and a positive control group (100 μL of culture medium containing 5-FU, with a final concentration of 50 μg / mL). Each group had 6 replicates, and the cells were cultured for another 48 hours. Then, MTT assay was performed: 20 μL of MTT solution (5 mg / mL) was added to each well, and the mixture was incubated at 37°C for 4 hours. The supernatant was carefully aspirated, and 150 μL of DMSO was added to each well. The mixture was shaken for 10 minutes to fully dissolve the crystals, and the absorbance (OD value) was measured at 490 nm using a microplate reader.

[0036] The formula for calculating the cell proliferation inhibition rate is: Cell proliferation inhibition rate (%) = [1 - (OD-treated group - OD blank) / (OD control group - OD blank)] × 100% MTT assay was used to detect the inhibitory effect on the proliferation of human hepatocellular carcinoma HepG2 cells (48 h, concentration 100 μg / mL).

[0037]

[0038] Note: Different letters in the superscript of data in the same column indicate significant differences (P<0.05).

[0039] Test Example 2: Mouse Xenograft Inhibition Experiment SPF-grade male Kunming mice, weighing 18-22 g, were kept in an SPF-grade animal room (temperature 22±2℃, humidity 50±10%, 12-hour light-dark cycle) with free access to water and food.

[0040] Mice with H22 hepatocellular carcinoma cells were passaged and sacrificed 7-8 days post-inoculation when the ascites was abundant. The ascites fluid was aseptically aspirated, diluted with physiological saline, and the cell concentration was adjusted to 2 × 10⁻⁶ cells after counting using a hemocytometer. 7 Cells / mL (confirmed viability >95% by trypan blue staining), followed by subcutaneous inoculation of 0.2 mL of cell suspension (containing 4 × 10⁶ cells / mL) into the right axilla of Kunming mice. 6 (1 cell), and mice were randomly divided into groups 24 hours after inoculation to ensure that there was no significant difference in the initial weight of each group.

[0041] The following groups were set up: a model control group (physiological saline by gavage, 0.2 mL / 10 g), multiple example groups (Examples 1-3, all 200 mg / kg by gavage), multiple comparative groups (Comparative Examples 1-5, all 200 mg / kg by gavage), and a positive control group (cyclophosphamide 20 mg / kg, intraperitoneal injection), with 10 mice in each group. Starting from the first day after inoculation, the mice were administered the drug once a day at a fixed time (9:00-10:00), with the gavage volume calculated at 0.2 mL / 10 g body weight, for 14 consecutive days.

[0042] Observe the general condition of the mice daily (mentality, activity, coat color, food and water intake, etc.) and record mortality; measure body weight every 3 days to calculate the rate of change; starting from day 5 post-inoculation, measure the long diameter (a) and short diameter (b) of the tumor every 3 days using calipers, and calculate the value using the formula V=(a×b). 2 The tumor volume was calculated as () / 2. Twenty-four hours after the last administration (day 15), mice were fasted but allowed free access to water for 12 hours. Then, 1 mL of blood was collected from the orbital venous plexus and placed in an anticoagulant tube. Mice were anesthetized with an intraperitoneal injection of 10% chloral hydrate (0.3 mL / 10g), euthanized by cervical dislocation, and the tumor tissue was quickly dissected. The intact tumor was then removed, blood was aspirated, and the tumor was weighed. The tumor inhibition rate was calculated. Tumor inhibition rate (%) = (average tumor weight in the model group - average tumor weight in the treatment group) / average tumor weight in the model group × 100%.

[0043]

[0044] At the same time, the pancreas and thymus were completely dissected, weighed, and their index was calculated.

[0045] Note: Different letters in the superscript of data in the same column indicate significant differences (P<0.05).

[0046] Test Example 3: Combined Chemotherapy Enhancement and Toxicity Reduction Experiment H22 tumor-bearing mouse models were established using the same method as in Test Example 2. The grouping and administration regimens were as follows (n=12 per group): 1) Model control group (saline gavage + intraperitoneal injection); 2) DDP-only group (cisplatin 2 mg / kg intraperitoneal injection, every other day); 3) Example 2-only group (Example 2 sample 200 mg / kg gavage, once daily); 4) Example 1 + DDP group; 5) Example 2 + DDP group; 6) Example 3 + DDP group; 7) Comparative Example 1 + DDP group; 8) Comparative Example 2 + DDP group; 9) G-CSF + DDP positive control group (G-CSF 50 μg / kg subcutaneous injection, once daily + cisplatin 2 mg / kg intraperitoneal injection, every other day). All groups were administered medication starting from day 1 post-inoculation for 14 consecutive days, with the administration sequence being gavage followed by injection, 2 hours apart.

[0047] The method for evaluating the anti-tumor effect is the same as that for test case 2.

[0048] Blood samples were collected via the orbital rim on days 7 and 14, and white blood cell counts (WBCs) were measured using a fully automated hematology analyzer. Serum was collected via the orbital rim 24 hours after the last administration, and liver function indicators (alanine aminotransferase ALT) and kidney function indicators (creatinine Cr) were measured using a fully automated biochemical analyzer.

[0049]

[0050] Note: Different letters in the superscript of data in the same column indicate significant differences (P<0.05).

[0051] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

Claims

1. An antitumor composition, characterized in that, By weight, it includes the following raw materials: Ginsenoside Rg3 18-22 parts, ginsenoside CK 3-7 parts, Ganoderma lucidum polysaccharide 28-32 parts, Astragalus polysaccharide 23-27 parts, Lentinan 8-12 parts, Panax notoginseng total saponins 16-20 parts, resveratrol 9-11 parts, curcumin 13-17 parts, dihydromyricetin 6-10 parts, matrine 7-9 parts; The ratio of the total mass of Ganoderma lucidum polysaccharides and Astragalus polysaccharides to the total mass of ginsenoside Rg3 and Panax notoginseng total saponins is (1.3-1.6):

1.

2. The antitumor composition according to claim 1, characterized in that, By weight, it includes the following raw materials: Ginsenoside Rg3 20 parts, ginsenoside CK 5 parts, Ganoderma lucidum polysaccharide 30 parts, Astragalus polysaccharide 25 parts, Lentinus edodes polysaccharide 10 parts, Panax notoginseng total saponins 18 parts, resveratrol 10 parts, curcumin 15 parts, dihydromyricetin 8 parts, matrine 8 parts.

3. The antitumor composition according to claim 1, characterized in that, The ratio of the total mass of Ganoderma lucidum polysaccharides and Astragalus polysaccharides to the total mass of ginsenoside Rg3 and Panax notoginseng total saponins is 1.45:

1.

4. The antitumor composition according to any one of claims 1-3, characterized in that, The composition includes an appropriate amount of pharmaceutical excipients.

5. The antitumor composition according to any one of claims 1-4, characterized in that, The composition is available in the form of powder, capsule, tablet or injection.

6. A method for preparing the antitumor composition according to any one of claims 1-5, characterized in that, Includes the following steps: Weigh the raw materials according to the proportions, mix them evenly, and obtain the antitumor composition.

7. The use of an antitumor composition in the preparation of antitumor drugs or in combination with chemotherapy drugs to enhance efficacy and reduce toxicity, characterized in that, The antitumor composition is the antitumor composition according to any one of claims 1-5 or the antitumor composition prepared by the method according to claim 6.

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