Anti-tumor traditional Chinese medicine composition and preparation method thereof

Abstract

The invention provides an anti-tumor traditional Chinese medicine composition and a preparation method thereof, and belongs to the technical field of medicines. The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 1-2 parts of ginseng, 7-12 parts of astragalus membranaceus, 5-15 parts of subprostrate sophora, 5-8 parts of polyporus umbellatus, 7-10 parts of sculellaria barbata, 3-5 parts of stephania tetrandra, 2-4 parts of caulis spatholobi, 7-10 parts of scutellaria baicalensis, 7-12 parts of bighead atractylodes rhizome, 10-15 parts of longspur epimedium, 5-7 parts of capillary artemisia, 3-5 parts of glabrous sarcandra herb, 5-10 parts of oldenlandia diffusa and 3-5 parts of salvia miltiorrhiza. The prepared anti-tumor traditional Chinese medicine composition not only has better water solubility and good bioavailability, can effectively inhibit proliferation of tumor cells, induce apoptosis of cancer cells and inhibit growth of tumor stem cells, shows a better anti-tumor effect, but also has better anti-oxidation and anti-inflammatory activity, and is good in drug effect, free of toxic and side effects and high in safety.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to an anti-tumor Chinese medicine composition and a preparation method thereof. Background Art

[0002] Tumors are a large class of complex diseases that seriously threaten human health. At present, the etiology and pathology of tumors are still in the exploratory stage. The treatment methods adopted in clinical practice are mainly surgery, and the purpose of treatment is achieved by removing the tumor. However, the particularity of tumors is that tumor cells are metastatic and can metastasize in the body. This is the greatest harm of tumors to the human body. In order to prevent the metastasis of tumor cells, clinically, radiotherapy and chemotherapy are also carried out after surgical resection of the tumor in order to completely eliminate tumor cells and reduce the probability of tumor cell metastasis. Although radiotherapy and chemotherapy can kill cancer cells, because tumors coexist with normal tissues and organs and even form a complex whole together, normal tissue cells are damaged during radiotherapy and chemotherapy, inducing gastrointestinal reactions, bone marrow suppression, and liver, kidney and heart function damage, making the patient's body weaker, causing damage to the human immune system, and sometimes even losing more than the gain. In Western medicine, excluding surgery and only using drugs, the efficiency is not high, the adverse reactions are huge, and the complex pathogenesis of tumors makes the current choice of chemical drugs to treat tumors not the most ideal means. In other words, there is still an embarrassing situation in the treatment of tumors with chemical drugs. Conventional chemical drug treatments are difficult to prevent tumor cell metastasis and have significant toxic side effects, which is the main problem in current tumor treatment. Therefore, finding a drug that can effectively eliminate tumor cells with minimal toxic side effects is the goal to be pursued in the current treatment of tumor diseases.

[0003] Traditional Chinese medicine is a huge treasure house. The development of traditional Chinese medicine theory has laid a foundation for the treatment of tumors. The scrofula, goiter, breast abscess, carbuncle, sputum nucleus formed by phlegm coagulation and blood stasis described in traditional Chinese medicine theory are all causes of tumors. Traditional Chinese medicine has effective treatment methods and drugs for the above diseases. As we all know, each Chinese herbal medicine contains complex ingredients, ranging from a dozen to hundreds. It is worthwhile to screen reasonable traditional Chinese medicines to prepare anti-tumor drugs. In fact, many drugs prepared by traditional Chinese medicine have played an important role in the treatment of tumors. Different tumors have different etiologies and pathologies, and traditional Chinese medicine treatment will also have different basic principles. There should be a broad space for exploration in the treatment of cancer with traditional Chinese medicine. It is of great significance to find a more effective, safer and less toxic drug for the treatment of tumors. Summary of the invention

[0004] The purpose of the present invention is to provide an anti-tumor Chinese medicine composition and a preparation method thereof, which not only has good water solubility and good bioavailability, can effectively inhibit the proliferation of tumor cells, induce cancer cell apoptosis and inhibit the growth of tumor stem cells, showing good anti-tumor effect, but also has good antioxidant and anti-inflammatory activity, good efficacy, no toxic side effects and high safety.

[0005] The technical solution of the present invention is achieved in this way:

[0006] The invention provides an anti-tumor Chinese medicine composition, which is prepared from the following raw materials in parts by weight: 1-2 parts of ginseng, 7-12 parts of scutellaria baicalensis, 5-15 parts of sophora tonkinensis roots, 5-8 parts of umbellatus, 7-10 parts of scutellaria barbata, 3-5 parts of tetrandra, 2-4 parts of millettia reticulata, 7-10 parts of scutellaria baicalensis, 7-12 parts of atractylodes macrocephala, 10-15 parts of epimedium, 5-7 parts of capillaris, 3-5 parts of sarmentosa, 5-10 parts of oldenlandia diffusa and 3-5 parts of salvia miltiorrhiza.

[0007] As a further improvement of the present invention, the following steps are included:

[0008] S1. Wash ginseng, Sophora flavescens, Stephania tetrandra, and Scutellaria baicalensis separately, dry, slice, obtain Chinese medicine tablets, mix with water, heat to boiling for extraction, filter, repeat extraction 2-3 times, filter, combine the extracts, retain the filtered solids, concentrate the extracts, add ethanol for precipitation, filter, wash the solids, dry, obtain Chinese medicine polysaccharides, add ammonium sulfate to the filtrate for graded precipitation, wash the precipitate, freeze-dry, obtain Chinese medicine protein, dialyze the filtrate, dry, and obtain Chinese medicine water extract;

[0009] S2. The filtered solids in step S1 are mixed with ethanol, heated for extraction, filtered, and the extraction is repeated 2-3 times, filtered, the extracts are combined, concentrated, and purified with a macroporous resin to obtain an active extract;

[0010] S3. adding the active extract to toluene, adding Chinese medicine polysaccharides and a catalyst, heating to reflux and stirring to react, filtering, washing, and drying to obtain an active extract / polysaccharide complex;

[0011] S4. adding the active extract / polysaccharide complex to water, adding NHS and EDC·HCl, activating the reaction, adding the Chinese medicine protein, stirring the reaction, dialyzing, and freeze-drying to obtain the active extract / polysaccharide / protein complex;

[0012] S5. The Polyporus umbellatus, Scutellaria barbata, Millettia spatholobi, Scutellaria baicalensis, Atractylodes macrocephala, Epimedium, Artemisia capillaris, Sarcandra chinensis, Hedyotis diffusa, and Salvia miltiorrhiza were washed, dried, sliced, mixed with water, heated to boiling for extraction, filtered, and the extraction was repeated 2-3 times, filtered, the extracts were combined, and the extracts were dried to obtain a Chinese medicine extract;

[0013] S6. Evenly mix the active extract / polysaccharide / protein complex, the Chinese medicine extract, and the Chinese medicine water extract, and granulate to obtain an anti-tumor Chinese medicine composition.

[0014] As a further improvement of the present invention, the solid-liquid ratio of the Chinese medicine tablets and water in step S1 is 1:7-10 g / mL, the time for heating to boiling extraction is 3-5 hours, the ethanol is added to 70-80wt% of the ethanol content of the system, the precipitation time is 2-3 hours, and the dialysis bag pore size used in the dialysis is 3000-4000Da, and the time is 24-36 hours.

[0015] As a further improvement of the present invention, the solid-liquid ratio of the filtered solid to ethanol in step S2 is 1:3-5 g / mL, the temperature of the heating extraction is 70-80°C, the time is 2-4 hours, and the macroporous resin is selected from at least one of D101, AB-8, HP-20, D301, and DA201.

[0016] As a further improvement of the present invention, the mass ratio of the active extract, the Chinese medicine polysaccharide and the catalyst in step S3 is 10:4-7:0.5-1, the heating reflux stirring reaction time is 7-10h, and the catalyst is p-toluenesulfonic acid or concentrated sulfuric acid.

[0017] As a further improvement of the present invention, the mass ratio of the active extract / polysaccharide complex, NHS, EDC·HCl, and traditional Chinese medicine protein in step S4 is 12-15:4-6:3-5:5-7, the temperature of the activation reaction is 0-4°C, the time is 30-50min, the stirring reaction time is 10-12h, and the dialysis bag pore size used in the dialysis is 5-7KDa, and the time is 24-36h.

[0018] As a further improvement of the present invention, the time for heating to boiling extraction in step S5 is 2-4 hours, and the mass ratio of active extract / polysaccharide / protein complex, Chinese medicine extract and Chinese medicine water extract in step S6 is 15-20:7-10:3-5.

[0019] The present invention further protects a method for preparing an anti-tumor co-crystal, which comprises dissolving the above-mentioned active extract / polysaccharide / protein complex in water to obtain solution A; dissolving berberine in water to obtain solution B; and dissolving solanine in hot ethanol to obtain solution C; and uniformly mixing solution A, solution B and solution C, evaporating the solvent in an open manner, filtering, washing, and drying to obtain the anti-tumor co-crystal.

[0020] As a further improvement of the present invention, the mass ratio of the active extract / polysaccharide complex, berberine and solanine is 10-12:1-2:2-3, and the temperature of the hot ethanol is 60-70°C.

[0021] The present invention further protects an anti-tumor co-crystal prepared by the above preparation method.

[0022] The present invention has the following beneficial effects:

[0023] The Chinese medicine composition of the invention comprises ginseng, scutellaria baicalensis, sophora flavescens, umbellatus, scutellaria barbata, tetrandra stephania tetrandra, millettia spatholobi, scutellaria baicalensis, atractylodes macrocephala, epimedium, capillaris, scutellaria scabra, oldenlandia diffusa and salvia miltiorrhiza. After water extraction, alcohol precipitation is performed to obtain Chinese medicine polysaccharides, and ammonium sulfate graded precipitation is performed to obtain Chinese medicine proteins. The water extract contains rich saponin active components such as ginsenoside, astragaloside and icariin, as well as alkaloid active components, which effectively inhibit the proliferation of tumor cells, induce cancer cell apoptosis and inhibit the growth of tumor stem cells, and show good anti-tumor effect.

[0024] After the solids filtered from the traditional Chinese medicine are extracted with alcohol, the obtained alcohol extract is purified by macroporous resin to obtain active extracts such as flavonoids, triterpenes, and lactones with higher activity. However, these components have poor water solubility and poor efficacy. Therefore, the present invention further modifies the structure of the active extract and couples it with traditional Chinese medicine protein, which not only improves the water solubility, but also improves the efficacy and broadens the application range of anti-tumor, and has good antioxidant and anti-inflammatory effects. However, due to the introduction of protein, the product is prone to allergies and other problems. Therefore, the present invention further uses traditional Chinese medicine polysaccharide compounding, so that the polysaccharide structure hides the sites on the protein surface that are prone to allergic reactions, thereby improving the safety of the product, and further improving the water solubility and efficacy of the product. The obtained active extract / polysaccharide / protein complex has a better anti-tumor effect.

[0025] Solanine can effectively inhibit the growth and proliferation of tumor cells, and inhibit the growth of cancer cells by mediating the mutual regulation between miR-375-3p and long non-coding RNA-colon cancer transcription factor 1. It can also induce cell cycle arrest and promote the apoptosis of cancer cells. However, it is not easily soluble in water, has low bioavailability, and is not easy to exert its efficacy. Berberine can cause cell cycle arrest and autophagy, promote cell apoptosis and inhibit tumor cell angiogenesis, thereby achieving the purpose of inhibiting tumor cell activity.

[0026] The present invention mixes the prepared active extract / polysaccharide / protein complex with berberine and solanine to prepare a drug co-crystal, which not only improves the water solubility of the product and overcomes the disadvantage that solanine is not easily soluble in water, but also greatly improves the drug efficacy and the stability of the product, overcomes the problem of poor thermal stability of the prepared active extract / polysaccharide / protein complex, and the prepared drug co-crystal has good anti-tumor activity, stability and water solubility, good drug efficacy and good bioavailability.

[0027] The anti-tumor Chinese medicine composition prepared by the present invention not only has good water solubility and good bioavailability, can effectively inhibit the proliferation of tumor cells, induce cancer cell apoptosis and inhibit the growth of tumor stem cells, showing good anti-tumor effect, but also has good antioxidant and anti-inflammatory activities, good efficacy, no toxic side effects and high safety. DETAILED DESCRIPTION

[0028] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0029] Example 1

[0030] This embodiment provides a method for preparing an anti-tumor Chinese medicine composition, comprising the following steps:

[0031] S1. Wash 1g of ginseng, 5g of Sophora flavescens, 3g of Stephania tetrandra, and 7g of Scutellaria baicalensis respectively, dry them, slice them, and obtain Chinese medicine tablets, mix them with water, the solid-liquid ratio of the Chinese medicine tablets to water is 1:7g / mL, heat to boiling and extract for 3h, filter, repeat the extraction twice, filter, combine the extracts, retain the filtered solids, concentrate the extracts, add ethanol to 70wt% of the ethanol content of the system, precipitate for 2h, filter, wash the solids, dry them, obtain Chinese medicine polysaccharides, add ammonium sulfate to the filtrate for graded precipitation, wash the precipitate, freeze-dry, obtain Chinese medicine protein, and dialyze the filtrate for 24h using a dialysis bag with a pore size of 3000Da, and dry them to obtain Chinese medicine water extracts;

[0032] S2. Mix the filtered solids in step S1 with ethanol, wherein the solid-liquid ratio of the filtered solids to ethanol is 1:3 g / mL, heat to 70°C, extract for 2 hours, filter, repeat the extraction twice, filter, combine the extracts, concentrate, purify with macroporous resin HP-20, the eluent is a mixture of ethanol and water, collect the eluent with an ethanol concentration of 40-60wt%, and obtain an active extract;

[0033] S3. Add 10 g of the active extract to 200 mL of toluene, add 4 g of Chinese medicine polysaccharide and 0.5 g of p-toluenesulfonic acid, heat under reflux and stir to react for 7 h, filter, wash, and dry to obtain an active extract / polysaccharide complex;

[0034] S4. 12 g of active extract / polysaccharide complex was added to 500 mL of water, 4 g of NHS and 3 g of EDC·HCl were added, the reaction was activated at 0°C for 30 min, 5 g of Chinese medicine protein was added, the reaction was stirred for 10 h, dialyzed for 24 h using a dialysis bag with a pore size of 5 KDa, and freeze-dried to obtain an active extract / polysaccharide / protein complex;

[0035] S5. 5g of Poria, 7g of Scutellaria barbata, 2g of Millettia spatholobi, 7g of Scutellaria baicalensis, 7g of Atractylodes macrocephala, 10g of Epimedium, 5g of Capillaris, 3g of Sarcandra scutellariae, 5g of Hedyotis diffusa, and 3g of Salvia miltiorrhiza were washed, dried, sliced, mixed with 10 times the mass of water, heated to boiling for extraction h, filtered, and the extraction was repeated 2 times, filtered, the extracts were combined, and the extracts were dried to obtain a Chinese medicine extract;

[0036] S6. Evenly mix 15 g of active extract / polysaccharide / protein complex, 7 g of Chinese medicine extract, and 3 g of Chinese medicine water extract, and granulate to obtain an anti-tumor Chinese medicine composition.

[0037] Example 2

[0038] This embodiment provides a method for preparing an anti-tumor Chinese medicine composition, comprising the following steps:

[0039] S1. Wash 2g of ginseng, 15g of Sophora flavescens, 5g of Stephania tetrandra, and 10g of Scutellaria baicalensis respectively, dry them, slice them, and obtain Chinese medicine tablets, mix them with water, the solid-liquid ratio of the Chinese medicine tablets to water is 1:10g / mL, heat to boiling and extract for 5h, filter, repeat the extraction 3 times, filter, combine the extracts, retain the filtered solids, concentrate the extracts, add ethanol to 80wt% of the ethanol content of the system, precipitate for 3h, filter, wash the solids, dry them, obtain Chinese medicine polysaccharides, add ammonium sulfate to the filtrate for graded precipitation, wash the precipitate, freeze-dry, obtain Chinese medicine protein, and dialyze the filtrate for 36h using a dialysis bag with a pore size of 4000Da, and dry them to obtain Chinese medicine water extracts;

[0040] S2. Mix the filtered solids in step S1 with ethanol, wherein the solid-liquid ratio of the filtered solids to ethanol is 1:5 g / mL, heat to 80°C, extract for 4 hours, filter, repeat the extraction 3 times, filter, combine the extracts, concentrate, purify with macroporous resin AB-8, the eluent is a mixture of ethanol and water, collect the eluent with an ethanol concentration of 40-60wt%, and obtain an active extract;

[0041] S3. Add 10 g of the active extract to 200 mL of toluene, add 7 g of Chinese medicine polysaccharide and 1 g of concentrated sulfuric acid, heat under reflux and stir to react for 10 h, filter, wash, and dry to obtain an active extract / polysaccharide complex;

[0042] S4. 15 g of active extract / polysaccharide complex was added to 500 mL of water, 6 g of NHS and 5 g of EDC·HCl were added, the reaction was activated at 4°C for 50 min, 7 g of Chinese medicine protein was added, the reaction was stirred for 12 h, dialyzed for 36 h using a dialysis bag with a pore size of 7 KDa, and freeze-dried to obtain an active extract / polysaccharide / protein complex;

[0043] S5. Wash 8g of Polyporus, 10g of Scutellaria barbata, 4g of Millettia spatholobi, 12g of Scutellaria baicalensis, 12g of Atractylodes macrocephala, 15g of Epimedium, 7g of Artemisia capillaris, 5g of Radix scutellariae, 10g of Hedyotis diffusa, and 5g of Salvia miltiorrhiza separately, dry, slice, mix with 10 times the mass of water, heat to boiling and extract for 4h, filter, repeat the extraction 3 times, filter, combine the extracts, and dry the extracts to obtain a Chinese medicine extract;

[0044] S6. Evenly mix 20 g of active extract / polysaccharide / protein complex, 10 g of Chinese medicine extract, and 5 g of Chinese medicine water extract, and granulate to obtain an anti-tumor Chinese medicine composition.

[0045] Example 3

[0046] This embodiment provides a method for preparing an anti-tumor Chinese medicine composition, comprising the following steps:

[0047] S1. Wash 1.5g of ginseng, 10g of Sophora flavescens, 4g of Stephania tetrandra, and 8g of Scutellaria baicalensis, dry them, slice them, and obtain Chinese medicine tablets, mix them with water, the solid-liquid ratio of the Chinese medicine tablets to water is 1:8g / mL, heat to boiling and extract for 4h, filter, repeat the extraction 3 times, filter, combine the extracts, retain the filtered solids, concentrate the extracts, add ethanol to 75wt% of the ethanol content of the system, precipitate for 2.5h, filter, wash the solids, dry them, obtain Chinese medicine polysaccharides, add ammonium sulfate to the filtrate for graded precipitation, wash the precipitate, freeze-dry, obtain Chinese medicine protein, and dialyze the filtrate for 30h using a dialysis bag with a pore size of 3500Da, and dry them to obtain Chinese medicine water extracts;

[0048] S2. Mix the filtered solids in step S1 with ethanol, wherein the solid-liquid ratio of the filtered solids to ethanol is 1:4 g / mL, heat to 75°C, extract for 3 hours, filter, repeat the extraction 3 times, filter, combine the extracts, concentrate, purify with macroporous resin D101, the eluent is a mixture of ethanol and water, collect the eluent with an ethanol concentration of 40-60wt%, and obtain an active extract;

[0049] S3. Add 10 g of the active extract to 200 mL of toluene, add 5.5 g of Chinese medicine polysaccharide and 0.7 g of concentrated sulfuric acid, heat to reflux and stir to react for 8 h, filter, wash, and dry to obtain an active extract / polysaccharide complex;

[0050] S4. 13 g of active extract / polysaccharide complex was added to 500 mL of water, 5 g of NHS and 4 g of EDC·HCl were added, the reaction was activated at 2°C for 40 min, 6 g of Chinese medicine protein was added, the reaction was stirred for 11 h, dialyzed for 30 h using a dialysis bag with a pore size of 6 KDa, and freeze-dried to obtain an active extract / polysaccharide / protein complex;

[0051] S5. Wash 6g of Polyporus, 8g of Scutellaria barbata, 3g of Millettia spatholobi, 10g of Scutellaria baicalensis, 10g of Atractylodes macrocephala, 12g of Epimedium, 6g of Capillaris, 4g of Radix Artemisiae Scopariae, 7g of Hedyotis diffusa, and 4g of Salvia miltiorrhiza, dry them, slice them, mix them with 10 times the mass of water, heat them to boiling and extract for 3h, filter them, repeat the extraction 3 times, filter them, combine the extracts, and dry the extracts to obtain a Chinese medicine extract;

[0052] S6. Evenly mix 17 g of active extract / polysaccharide / protein complex, 8 g of Chinese medicine extract, and 4 g of Chinese medicine water extract, and granulate to obtain an anti-tumor Chinese medicine composition.

[0053] Comparative Example 1

[0054] Compared with Example 3, the difference is that step S3 is not performed.

[0055] The details are as follows:

[0056] S1. Wash 1.5g of ginseng, 10g of Sophora flavescens, 4g of Stephania tetrandra, and 8g of Scutellaria baicalensis, dry them, slice them, and obtain Chinese medicine tablets, mix them with water, the solid-liquid ratio of the Chinese medicine tablets to water is 1:8g / mL, heat to boiling and extract for 4h, filter, repeat the extraction 3 times, filter, combine the extracts, retain the filtered solids, concentrate the extracts, add ethanol to 75wt% of the ethanol content of the system, precipitate for 2.5h, filter, wash the solids, dry them, obtain Chinese medicine polysaccharides, add ammonium sulfate to the filtrate for graded precipitation, wash the precipitate, freeze-dry, obtain Chinese medicine protein, and dialyze the filtrate for 30h using a dialysis bag with a pore size of 3500Da, and dry them to obtain Chinese medicine water extracts;

[0057] S2. Mix the filtered solids in step S1 with ethanol, wherein the solid-liquid ratio of the filtered solids to ethanol is 1:4 g / mL, heat to 75°C, extract for 3 hours, filter, repeat the extraction 3 times, filter, combine the extracts, concentrate, purify with macroporous resin D101, the eluent is a mixture of ethanol and water, collect the eluent with an ethanol concentration of 40-60wt%, and obtain an active extract;

[0058] S3. 13 g of active extract was added to 500 mL of water, 5 g of NHS and 4 g of EDC·HCl were added, the reaction was activated at 2°C for 40 min, 6 g of Chinese medicine protein was added, the reaction was stirred for 11 h, dialyzed for 30 h using a dialysis bag with a pore size of 6 KDa, and freeze-dried to obtain an active extract / protein complex;

[0059] S4. Wash 6g of Polyporus, 8g of Scutellaria barbata, 3g of Millettia spatholobi, 10g of Scutellaria baicalensis, 10g of Atractylodes macrocephala, 12g of Epimedium, 6g of Capillaris, 4g of Radix Artemisiae Scopariae, 7g of Hedyotis diffusa, and 4g of Salvia miltiorrhiza, dry them, slice them, mix them with 10 times the mass of water, heat them to boiling and extract for 3h, filter them, repeat the extraction 3 times, filter them, combine the extracts, and dry the extracts to obtain a Chinese medicine extract;

[0060] S5. Evenly mix 17 g of active extract / protein complex, 8 g of Chinese medicine extract, and 4 g of Chinese medicine water extract, and granulate to obtain an anti-tumor Chinese medicine composition.

[0061] Comparative Example 2

[0062] Compared with Example 3, the difference is that step S4 is not performed.

[0063] The details are as follows:

[0064] S1. Wash 1.5g of ginseng, 10g of Sophora flavescens, 4g of Stephania tetrandra, and 8g of Scutellaria baicalensis, dry them, slice them, and obtain Chinese medicine tablets, mix them with water, the solid-liquid ratio of the Chinese medicine tablets to water is 1:8g / mL, heat to boiling and extract for 4h, filter, repeat the extraction 3 times, filter, combine the extracts, retain the filtered solids, concentrate the extracts, add ethanol to 75wt% of the ethanol content of the system, precipitate for 2.5h, filter, wash the solids, dry them, obtain Chinese medicine polysaccharides, add ammonium sulfate to the filtrate for graded precipitation, wash the precipitate, freeze-dry, obtain Chinese medicine protein, and dialyze the filtrate for 30h using a dialysis bag with a pore size of 3500Da, and dry them to obtain Chinese medicine water extracts;

[0065] S2. Mix the filtered solids in step S1 with ethanol, wherein the solid-liquid ratio of the filtered solids to ethanol is 1:4 g / mL, heat to 75°C, extract for 3 hours, filter, repeat the extraction 3 times, filter, combine the extracts, concentrate, purify with macroporous resin D101, the eluent is a mixture of ethanol and water, collect the eluent with an ethanol concentration of 40-60wt%, and obtain an active extract;

[0066] S3. Add 10 g of the active extract to 200 mL of toluene, add 5.5 g of Chinese medicine polysaccharide and 0.7 g of concentrated sulfuric acid, heat to reflux and stir to react for 8 h, filter, wash, and dry to obtain an active extract / polysaccharide complex;

[0067] S4. Wash 6g of Polyporus, 8g of Scutellaria barbata, 3g of Millettia spatholobi, 10g of Scutellaria baicalensis, 10g of Atractylodes macrocephala, 12g of Epimedium, 6g of Capillaris, 4g of Radix Artemisiae Scopariae, 7g of Hedyotis diffusa, and 4g of Salvia miltiorrhiza, dry them, slice them, mix them with 10 times the mass of water, heat them to boiling and extract for 3h, filter them, repeat the extraction 3 times, filter them, combine the extracts, and dry the extracts to obtain a Chinese medicine extract;

[0068] S5. Evenly mix 17 g of active extract / polysaccharide complex, 8 g of Chinese herbal extract, and 4 g of Chinese herbal water extract, and granulate to obtain an anti-tumor Chinese herbal composition.

[0069] Test Example 1

[0070] The anti-tumor Chinese medicine compositions prepared in Examples 1-3 of the present invention and Comparative Examples 1-2 were subjected to a solubility test to test the solubility of the compositions in 100 g of distilled water at room temperature. The results are shown in Table 1.

[0071] Table 1

[0072] Group Solubility (g / 100g) Example 1 11.2 Example 2 11.5 Example 3 12.0 Comparative Example 1 2.5 Comparative Example 2 7.6

[0073] It can be seen from the above table that the anti-tumor Chinese medicine compositions prepared in Examples 1-3 of the present invention are easily soluble in water.

[0074] Test Example 2

[0075] Weaned female BALB / c mice (4 weeks old) after adaptive feeding for 7 days were randomly divided into 4 groups according to body weight: normal control group, sensitized control group, Example 3 group and comparative example 2 group, 10 mice in each group. During the test period (0-42 days), each intervention group of mice was fed with an aqueous solution containing an anti-tumor Chinese medicine composition (10 mg / mL) instead of drinking water for feeding intervention every day, and the control group (including the normal control group and the sensitized control group) mice were fed with normal drinking water as needed. The sensitized control group and each intervention group of mice were gavaged with milk protein (20 mg / mouse) + cholera toxin (10 μg / mouse) on the 7th, 14th, 21st, 28th, and 35th days of the test, and the normal control group mice were gavaged with sterile saline; on the 42nd day of the test, all mice were gavaged with milk protein (50 mg / mouse).

[0076] Allergy symptom scoring Systemic allergy symptom scoring of mice: The specific scoring criteria are as follows: 0 points = no symptoms; 1 point = scratching around the nose and head; 2 points = swelling around the eyes and mouth, erect hair, reduced activity, and increased respiratory rate; 3 points = wheezing, labored breathing, and cyanosis around the mouth and tail; 4 points = trembling, convulsions, and no activity after stimulation; 5 points = death.

[0077] The results are shown in Table 2.

[0078] Table 2

[0079] Group Allergy symptom score Normal control group 0 Sensitized control group 3.21 Example 3 Group 1.02 Comparative Example 2 4.27

[0080] It can be seen from the above table that the anti-tumor Chinese medicine composition prepared in Example 3 of the present invention can alleviate allergic symptoms to a certain extent, while the anti-tumor Chinese medicine composition prepared in Comparative Example 2 aggravates the allergic symptoms.

[0081] Example 4

[0082] This embodiment provides an anti-tumor co-crystal, and the preparation method is as follows:

[0083] 10 g of the active extract / polysaccharide / protein complex obtained in step S4 of Example 1 was dissolved in 200 mL of water to obtain solution A; 1 g of berberine was dissolved in 200 mL of water to obtain solution B; 2 g of solanine was dissolved in 200 mL of hot ethanol at 60° C. to obtain solution C; solution A, solution B and solution C were mixed evenly, the solvent was evaporated in an open state for 7 days, filtered, washed, and dried to obtain anti-tumor cocrystals.

[0084] Example 5

[0085] This embodiment provides an anti-tumor co-crystal, and the preparation method is as follows:

[0086] 12 g of the active extract / polysaccharide / protein complex obtained in step S4 of Example 2 was dissolved in 200 mL of water to obtain solution A; 2 g of berberine was dissolved in 200 mL of water to obtain solution B; 3 g of solanine was dissolved in 200 mL of 70° C. hot ethanol to obtain solution C; solution A, solution B and solution C were mixed evenly, the solvent was evaporated in an open state for 7 days, filtered, washed, and dried to obtain anti-tumor cocrystals.

[0087] Example 6

[0088] This embodiment provides an anti-tumor co-crystal, and the preparation method is as follows:

[0089] 11 g of the active extract / polysaccharide / protein complex obtained in step S4 of Example 3 was dissolved in 200 mL of water to obtain solution A; 1.5 g of berberine was dissolved in 200 mL of water to obtain solution B; 2.5 g of solanine was dissolved in 200 mL of hot ethanol at 65° C. to obtain solution C; solution A, solution B and solution C were mixed evenly, the solvent was evaporated in an open state for 7 days, filtered, washed, and dried to obtain anti-tumor cocrystals.

[0090] Comparative Example 3

[0091] Compared with Example 6, the difference is that no active extract / polysaccharide / protein complex is added.

[0092] The details are as follows:

[0093] 1.5 g of berberine was dissolved in 200 mL of water to obtain solution B, and 2.5 g of solanine was dissolved in 200 mL of 65° C. hot ethanol to obtain solution C. Solution B and solution C were mixed evenly, the solvent was evaporated in an open air for 7 days, filtered, washed, and dried to obtain antitumor cocrystals.

[0094] Comparative Example 4

[0095] Compared with Example 6, the difference is that berberine is not added.

[0096] The details are as follows:

[0097] 11 g of the active extract / polysaccharide / protein complex obtained in step S4 of Example 3 was dissolved in 200 mL of water to obtain solution A; 2.5 g of solanine was dissolved in 200 mL of hot ethanol at 65° C. to obtain solution C; solution A and solution C were mixed evenly, the solvent was evaporated in an open state for 7 days, filtered, washed, and dried to obtain anti-tumor cocrystals.

[0098] Comparative Example 5

[0099] Compared with Example 6, the difference is that solanine is not added.

[0100] The details are as follows:

[0101] 11 g of the active extract / polysaccharide / protein complex obtained in step S4 of Example 3 was dissolved in 200 mL of water to obtain solution A; 1.5 g of berberine was dissolved in 200 mL of water to obtain solution B. Solution A and solution B were mixed evenly, the solvent was evaporated in an open state for 7 days, filtered, washed, and dried to obtain anti-tumor cocrystals.

[0102] Test Example 3

[0103] SPF female C57BL / 6 mice, 6 weeks old, were fed adaptively for 7 days before the experiment. The right side of the abdomen of the mice was shaved and inoculated subcutaneously with 5×10 5 The cell fluid of mouse melanoma cells B16F10 was purchased from the cell bank of the Chinese Academy of Sciences. The cells were cultured in RMPI1640 culture medium containing 1% penicillin-streptomycin and 10% FBS. When the tumor diameter was about 0.5 cm, the mice with uniform tumor growth were selected and randomly divided into a control group and a treatment group, with 5 mice in each group. The treatment group was injected intratumorally with 1 mg / mL of the anti-tumor Chinese medicine composition prepared in Example 1-6 or Comparative Example 1-5 or 2 mg / mouse of the anti-tumor co-crystal, once every two days; the control group was injected intratumorally with an equal volume of PBS solution. After 15 days, the tumor mass was peeled off and weighed, and the tumor inhibition rate (%) was calculated. The results are shown in Table 3.

[0104] Tumor inhibition rate (%) = (control group tumor mass weight - drug administration group tumor mass weight) / control group tumor mass weight × 100%

[0105] Table 3

[0106]

[0107]

[0108] It can be seen from the above table that the anti-tumor Chinese medicine composition prepared in Examples 1-3 or the anti-tumor co-crystals prepared in Examples 4-6 of the present invention have good anti-tumor effects.

[0109] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. An anti-tumor Chinese medicine composition, characterized in that: The invention is prepared from the following raw materials by weight: 1-2 parts of ginseng, 7-12 parts of stilbene, 5-15 parts of sophora flavescens, 5-8 parts of umbellatus, 7-10 parts of scutellaria barbata, 3-5 parts of tetrandra, 2-4 parts of millettia reticulata, 7-10 parts of scutellaria baicalensis, 7-12 parts of atractylodes macrocephala, 10-15 parts of epimedium, 5-7 parts of capillaris, 3-5 parts of sarmentosa, 5-10 parts of oldenlandia diffusa and 3-5 parts of salvia miltiorrhiza.

2. A method for preparing the anti-tumor Chinese medicine composition according to claim 1, characterized in that: The following steps are involved: S1. Wash ginseng, Sophora flavescens, Stephania tetrandra, and Scutellaria baicalensis separately, dry, slice, obtain Chinese medicine tablets, mix with water, heat to boiling for extraction, filter, repeat extraction 2-3 times, filter, combine the extracts, retain the filtered solids, concentrate the extracts, add ethanol for precipitation, filter, wash the solids, dry, obtain Chinese medicine polysaccharides, add ammonium sulfate to the filtrate for graded precipitation, wash the precipitate, freeze-dry, obtain Chinese medicine protein, dialyze the filtrate, dry, and obtain Chinese medicine water extract; S2. The filtered solids in step S1 are mixed with ethanol, heated for extraction, filtered, and the extraction is repeated 2-3 times, filtered, the extracts are combined, concentrated, and purified with a macroporous resin to obtain an active extract; S3. adding the active extract to toluene, adding Chinese medicine polysaccharides and a catalyst, heating to reflux and stirring to react, filtering, washing, and drying to obtain an active extract / polysaccharide complex; S4. adding the active extract / polysaccharide complex to water, adding NHS and EDC·HCl, activating the reaction, adding the Chinese medicine protein, stirring the reaction, dialyzing, and freeze-drying to obtain the active extract / polysaccharide / protein complex; S5. The Polyporus umbellatus, Scutellaria barbata, Millettia spatholobi, Scutellaria baicalensis, Atractylodes macrocephala, Epimedium, Artemisia capillaris, Sarcandra chinensis, Hedyotis diffusa, and Salvia miltiorrhiza were washed, dried, sliced, mixed with water, heated to boiling for extraction, filtered, and the extraction was repeated 2-3 times, filtered, the extracts were combined, and the extracts were dried to obtain a Chinese medicine extract; S6. Evenly mix the active extract / polysaccharide / protein complex, the Chinese medicine extract, and the Chinese medicine water extract, and granulate to obtain an anti-tumor Chinese medicine composition.

3. The preparation method according to claim 2, characterized in that: The solid-liquid ratio of the Chinese medicine tablets and water in step S1 is 1:7-10 g / mL, the time for heating to boiling extraction is 3-5 hours, the ethanol is added to 70-80wt% of the ethanol content of the system, the precipitation time is 2-3 hours, and the dialysis bag pore size used in the dialysis is 3000-4000Da, and the time is 24-36 hours.

4. The preparation method according to claim 2, characterized in that: The solid-liquid ratio of the filtered solid to ethanol in step S2 is 1:3-5 g / mL, the temperature of the heating extraction is 70-80°C, the time is 2-4 hours, and the macroporous resin is selected from at least one of D101, AB-8, HP-20, D301, and DA201.

5. The preparation method according to claim 2, characterized in that: In step S3, the mass ratio of the active extract, the Chinese medicine polysaccharide and the catalyst is 10:4-7:0.5-1, the heating reflux stirring reaction time is 7-10h, and the catalyst is p-toluenesulfonic acid or concentrated sulfuric acid.

6. The preparation method according to claim 2, characterized in that: In step S4, the mass ratio of the active extract / polysaccharide complex, NHS, EDC·HCl, and Chinese medicine protein is 12-15:4-6:3-5:5-7, the temperature of the activation reaction is 0-4°C, the time is 30-50min, the stirring reaction time is 10-12h, the dialysis bag pore size used in the dialysis is 5-7KDa, and the time is 24-36h.

7. The preparation method according to claim 2, characterized in that: The time for heating to boiling extraction in step S5 is 2-4 hours, and the mass ratio of active extract / polysaccharide / protein complex, Chinese medicine extract and Chinese medicine water extract in step S6 is 15-20:7-10:3-5.

8. A method for preparing an anti-tumor co-crystal, characterized in that: The active extract / polysaccharide / protein complex obtained in step S5 of claim 2 is dissolved in water to obtain solution A; berberine is dissolved in water to obtain solution B, and solanine is dissolved in hot ethanol to obtain solution C; solution A, solution B and solution C are mixed evenly, the solvent is evaporated in an open manner, filtered, washed, and dried to obtain anti-tumor co-crystals.

9. The preparation method according to claim 8, characterized in that: The mass ratio of the active extract / polysaccharide complex, berberine and solanine is 10-12:1-2:2-3, and the temperature of the hot ethanol is 60-70°C.

10. An anti-tumor co-crystal obtained by the preparation method according to claim 8 or 9.

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