Traditional Chinese medicine compound effective active ingredient preparation for resisting colorectal cancer and preparation method thereof

Abstract

The application discloses a Chinese medicine compound effective active ingredient preparation for resisting colorectal cancer and a preparation method thereof. The preparation is prepared from the following Chinese medicine raw materials in equal weight parts: Smilax riparia, Rabdosia rubescens, Oldenlandia diffusa and Centella asiatica. The active ingredient extraction method comprises the following steps: (1) immersing the Chinese medicine raw materials in 90% ethanol for extraction; (2) adding chitosan into the extraction liquid, stirring, standing for precipitation, solid-liquid separation, and obtaining a filtrate; (3) adjusting the pH value, standing for precipitation, solid-liquid separation, and obtaining a filtrate; (4) eluting the obtained filtrate on a macroporous adsorption resin D4020; and (5) performing vacuum concentration and drying on the obtained eluate, and then crushing the eluate into fine powder, thereby obtaining the preparation. Compared with the existing compound Smilax three herb soup, the preparation has an inhibition rate on colon cancer HCT116 and DIFI cells which is increased by more than 2 times, and is more than 90%.

Description

TECHNICAL FIELD

[0001] The present application relates to an anti-tumor compound Chinese herbal medicine preparation, in particular to an anti-colorectal cancer Chinese herbal medicine compound effective ingredient preparation and a preparation method thereof. BACKGROUND

[0002] The existing compound Goutengsan three herb soup, the main medicine includes Goutengsan, Henlingcao, Baihuasheshecao, Jixuecao, which is derived from clinical experience prescription, and the ratio is equal to the ratio. When used for the treatment and prevention of colorectal cancer, it is suggested that the prescription has significant anti-tumor effect (see Cao Liujian et al. Effect of compound Goutengsan on quality of life and immune function of patients with colorectal cancer after chemotherapy [J]. Chinese Journal of Cancer Prevention and Treatment, 2020, 27 (S1): 104-105; He Benshou et al. Observation of compound Goutengsan on adverse reactions and clinical efficacy of postoperative chemotherapy in patients with colorectal cancer [J]. Chinese Journal of Clinical Research, 2021, 13 (19): 13-15.) But what is the effective active ingredient for anti-colorectal cancer is not clear.

[0003] At present, the research on the effective active ingredients of compound Goutengsan three herb soup is limited to the research on the effective active ingredients of single medicine. The research on the extraction method of their effective active ingredients is not deep enough and cannot be applied in industry. The following will be introduced respectively.

[0004] Goutengsan is the dried root of Actinidia Chinensis Planch. or Actinidia arguta (Sieb & Zucc.) Planch. ex Miq. of the Actinidiaceae Actinidia genus, which has the effects of clearing heat and resolving toxins, promoting blood circulation and removing dampness, and contains triterpenoids, flavonoids, anthraquinones, steroids, alkaloids, and phenylpropanoids. Some people have found that it has anti-cancer effect (see Chen Dexuan et al. Anti-tumor plant medicine Goutengsan, Chinese Herbal Medicine, 2004, 27 (2): 86-87), and its ethanol extract can significantly induce apoptosis of colorectal cancer cells HCT15 and LoVo at a concentration of 50 μg / ml. Some people have studied its active ingredients and found that the triterpenoid ursolic acid contained in it can significantly induce apoptosis of colorectal cancer cells HCT116 (see Li Yongjing et al. Effect and mechanism of ursolic acid on reducing cholesterol content and inducing apoptosis of colorectal cancer cells [J]. Chinese Journal of Integrated Traditional Chinese and Western Medicine for Digestive Diseases, 2023, 31 (04): 266-273.

[0005] Rabdosia rubescens (Hemsl.) H. Hara is the dry aboveground whole grass of the Lamiaceae Xiangcha Suomi, with anti-tumor, anti-bacterial and anti-inflammatory, anti-oxidation and other pharmacological effects. The kopsane type of ring diterpenoids, oridonin, is one of the main active ingredients, with good anti-tumor activity and becoming the research hotspot of anti-tumor drugs today. At dosages of 4, 8 and 12 μg / mL, it can significantly inhibit the growth of colon cancer LoVo cells and induce apoptosis of colon cancer LoVo cells (Long J, Bu HQ, Liu DL. Experimental study on the effect of oridonin on the proliferation and EMT of colon cancer LOVO cells [J]. Practical Oncology, 2019, 34(04): 309-315). Oridonin also has an anti-proliferative effect on colon cancer SW620 cells and can induce apoptosis of colon cancer SW620 cells, increase the total level and phosphorylation level of p53, and reduce the growth rate of mouse tumors (see Liu RX, Ma Y, Hu XL, et al. Anticancer effects of oridoninon colon cancer are mediated via BMP7 / p38 MAPK / p53 signaling. Int J Oncol. 2018; 53(5): 2091-2101).

[0006] Hedyotis diffusa Willd is the dry whole grass of the Rubiaceae Hedyotis plant, with the effects of clearing heat and resolving toxicity, relieving pain and resolving nodules, and diuresis and dampness, mainly containing iridoid compounds, acid-containing compounds, anthraquinone compounds, volatile components, phenylpropanoids, coumarins and polysaccharides. It is a common drug in traditional Chinese medicine anti-tumor compound therapy, with anti-tumor activity including inhibition of tumor cell proliferation, inhibition of tumor angiogenesis, induction of tumor cell apoptosis, regulation of host immune response and protective autophagy, and is widely used in the clinical treatment of digestive system tumors. Hedyotis diffusa extract polysaccharide (50 μg / ml) and total flavonoids (5 μg / ml) have an inhibitory effect on colon cancer HT-29 cells, promote apoptosis, and inhibit the migration of colon cancer HT-29 cells (Xie DB, Guo YT. Inhibitory effect of Hedyotis diffusa extract polysaccharide and total flavonoids on colon cancer HT-29 cells [J]. China Medicine and Clinic, 2021, 21(19): 3217-3220). However, so far, there is no literature report on the separation method and improved preparation dosage form of its components.

[0007] Centella asiatica is the whole herb of the plant Centella asiatica, belonging to the Apiaceae family. It possesses the effects of clearing heat and dampness, promoting blood circulation and stopping bleeding, and detoxifying and reducing swelling. Total asiatic acid and asiaticoside are its main active ingredients. A asiaticoside concentration of 250 μg / mL significantly promoted apoptosis in colon cancer CT26 cells, exhibiting concentration-dependent apoptosis. A asiaticoside concentrations of 50 mg / kg and 100 mg / kg significantly inhibited tumorigenesis in mouse tumor cells (see Zhang Hua. Aasiaticoside induces apoptosis in colon cancer cells through immunomodulation and the NF-κB pathway [D]. Wuhan University, 2015.).

[0008] In summary, to date, there are no literature reports on the extraction and separation methods and formulation techniques of compound active ingredients composed of Actinidia chinensis root, Rabdosia rubescens, Hedyotis diffusa, and Centella asiatica. Summary of the Invention

[0009] The technical problem to be solved by the present invention is to provide a compound preparation of effective active ingredients of traditional Chinese medicine for colorectal cancer, composed of Actinidia chinensis root, Rabdosia rubescens, Hedyotis diffusa, and Centella asiatica, which is easy to operate, low in cost, and has good inhibitory and killing effects on colorectal cancer cells, and the preparation method thereof.

[0010] The technical solution adopted by this invention to solve its technical problem is a compound preparation of effective active ingredients of traditional Chinese medicine for colorectal cancer, made from the following parts by weight of traditional Chinese medicine raw materials: 1-3 parts of *Actinidia chinensis* root, 1-3 parts of *Rabdosia rubescens*, 1-3 parts of *Hedyotis diffusa*, and 1-3 parts of *Centella asiatica*; the extraction method of the effective active ingredients of the compound of traditional Chinese medicine raw materials *Actinidia chinensis* root, *Rabdosia rubescens*, *Hedyotis diffusa*, and *Centella asiatica* includes the following steps:

[0011] (1) Mix the raw medicinal materials of *Actinidia chinensis* root, *Rhizoma Scutellariae*, *Hedyotis diffusa*, and *Centella asiatica* or their pulverized parts, and extract them with a 90% ethanol solution (hereinafter referred to as "90% ethanol") for 1-2 hours. Filter, centrifuge to remove the precipitate, and retain the extract. Repeat the extraction 2-3 times and combine the extracts.

[0012] (2) Add chitosan to the extract obtained in step (1), stir, let it stand to precipitate, filter or centrifuge to remove the precipitate, and obtain filtrate or supernatant.

[0013] (3) Add pH adjuster to the filtrate or supernatant obtained in step (2) to adjust the pH value, let it stand to precipitate, filter or centrifuge to remove the precipitate, and obtain filtrate or supernatant.

[0014] (4) Load the filtrate or supernatant obtained in step (3) onto macroporous adsorption resin D4020, and then elute with ethanol solution to obtain eluent.

[0015] (5) the eluate obtained in step (4) is concentrated under reduced pressure to obtain a decoction, and the decoction is dried and then crushed into fine powder to obtain the anti-colorectal cancer Chinese herbal medicine compound active ingredient preparation crude drug (hereinafter sometimes referred to as the crude drug of the present application).

[0016] The crude drug of the present application can be added to a medically permissible excipient to prepare various different dosage forms such as granules and capsules for convenient clinical use.

[0017] Further, in step (1), the mass ratio of the crude Chinese medicine to 90% ethanol is 1:10 for the first time, 1:8 for the second time, and 1:6 for the third time in the 90% ethanol extraction; the extract can be concentrated or not concentrated and used in subsequent steps.

[0018] Further, in step (2), the amount of chitosan added is 0.1-1% by weight based on the weight of the crude Chinese medicine.

[0019] Further, in step (3), the pH value is adjusted to 2-5.

[0020] Further, the pH value adjusting agent is an acidic reagent such as hydrochloric acid or acetic acid.

[0021] Further, in step (4), the type of macroporous adsorption resin is D4020.

[0022] Further, in step (5), the drying process is one of vacuum drying, spray drying, freeze drying, and other drying methods.

[0023] Usage and dosage: granules: specifications: 3g / bag, 1 bag each time, decoction with boiling water, 2 times a day; capsules: specifications: 0.25g / pill, 2 pills each time, oral, 2 times a day.

[0024] The present application has the following advantages: (1) 90% ethanol is used for extraction for 2-3 times, each time for 1-2 hours, which is beneficial to improve the transfer rate (extraction rate) of the effective components in the raw medicinal materials as much as possible; (2) chitosan is added to the extraction liquid for precipitation operation, filtration, centrifugation and removal of the precipitate, which avoids the loss caused by incomplete precipitation of the ethanol precipitation method and the additional steps of recovering more ethanol; (3) the acid insoluble substances are removed by adjusting the acid method, which is beneficial to improve the adsorption efficiency of the subsequent macroporous adsorption resin on the effective active ingredients; (4) the macroporous adsorption resin D4020 is used to purify and refine the effective active ingredients in the extraction liquid, which can effectively purify the effective parts of the Chinese medicinal materials of Radix Tetrastigme, Rabdosia rubescens, Oldenlandia diffusa and Adiantum capillus-veneris (the content of the pentacyclic triterpenoid components is more than 80%, and the content of the flavonoid components is more than 60%), and is convenient for industrial production; (5) the effective active ingredients of the compound of Radix Tetrastigme, Rabdosia rubescens, Oldenlandia diffusa and Adiantum capillus-veneris are extracted to prepare a preparation, which can effectively improve the bioavailability and improve the drug efficacy: compared with the existing compound of Radix Tetrastigme, Rabdosia rubescens and Oldenlandia diffusa, the inhibition rate of the preparation on the HCT116 and DIFI cells of colon cancer is increased by more than 2 times, which is more than 90%; (6) the present application fills the gap of the existing anti-tumor compound of Radix Tetrastigme, Rabdosia rubescens, Oldenlandia diffusa and Adiantum capillus-veneris, and develops a new type of anti-tumor Chinese medicinal compound effective component preparation. DETAILED DESCRIPTION

[0025] The present application will be further described in detail below in combination with examples. Example 1

[0026] (1) 2 kg of Radix Tetrastigme, 1 kg of Rabdosia rubescens, 1 kg of Oldenlandia diffusa and 1 kg of Adiantum capillus-veneris are mixed and crushed into coarse powder, 10 times of 90% ethanol is added for heating reflux, and the time is counted for 90 minutes; filtration is performed, 8 times of 90% ethanol is added to the filter residue for heating reflux, and the time is counted for 60 minutes; filtration is performed, 6 times of 90% ethanol is added to the filter residue for heating reflux, and the time is counted for 60 minutes; filtration is performed; the three times of filtrates are combined, and the ethanol is recovered by vacuum concentration; the concentration is continued until the hot specific gravity is 1.05; (2) 5 g of chitosan is added, stirred, and precipitated, and the precipitate is removed by filtration or centrifugation to obtain the filtrate or supernatant; (3) the pH value is adjusted to 2 by using 10% hydrochloric acid, stirred uniformly, and the supernatant is obtained by centrifugation; (4) the supernatant is passed through a macroporous adsorption resin D4020 column, and then eluted with an ethanol solution to obtain an eluate; (5) the eluate is concentrated under reduced pressure, the vacuum degree is 0.05 Mpa, the extract is obtained, the extract is vacuum freeze-dried, the vacuum degree is greater than 0.02 Mpa, and the drying time is 4 hours; the extract is crushed into fine powder (the anti-colorectal cancer Chinese herbal compound effective active ingredient preparation of the present application, the same below), weighed, and 115 g is obtained.

[0027] The content of the pentacyclic triterpenoids and flavonoids in the dry product is 88% and 66% respectively, which is determined by Ultra Performance Liquid Chromatography (UPLC). Example 2

[0028] (1) Take 4 Kg of the roots of Actinidia deliciosa, 1 Kg of Rabdosia rubescens, 1 Kg of Oldenlandia diffusa and 1 Kg of Centella asiatica, crush them into coarse powder, add 10 times of 90% ethanol, heat and reflux for 90 minutes, filter, add 8 times of 90% ethanol to the residue, heat and reflux for 60 minutes, filter, add 6 times of 90% ethanol to the residue, heat and reflux for 60 minutes, combine the three filtrates, vacuum concentrate and recover ethanol, continue to concentrate to a hot specific gravity of 1.05; (2) add 5 g of chitosan, stir, stand, centrifuge, remove the precipitate, and obtain the supernatant; (3) adjust the pH of the supernatant to 3 with 10% hydrochloric acid, stand, centrifuge, remove the precipitate, and obtain the supernatant; (4) pass the supernatant through a macroporous adsorption resin D4020, elute with dilute hydrochloric acid, and obtain the eluate; (5) reduce the pressure to concentrate the eluate, the vacuum degree is 0.05 Mpa, vacuum freeze dry the extract, the vacuum degree is 0.02 Mpa, and the drying time is 4 hours; and a total of 109 g of dry product is prepared.

[0029] The content of the pentacyclic triterpenoids and flavonoids in the dry product is 91% and 69% respectively, which is determined by UPLC. Example 3

[0030] (1) Take 3 Kg of the roots of Actinidia deliciosa, 2 Kg of Rabdosia rubescens, 1 Kg of Oldenlandia diffusa and 1 Kg of Centella asiatica, crush them into coarse powder, add 10 times of 90% ethanol, heat and reflux for 90 minutes; filter, add 8 times of 90% ethanol to the residue, heat and reflux for 60 minutes; filter, add 6 times of 90% ethanol to the residue, heat and reflux for 60 minutes, combine the three filtrates, vacuum concentrate and recover ethanol, continue to concentrate to a hot specific gravity of 1.05; (2) add 5 g of chitosan, stir, stand, centrifuge, remove the precipitate, and obtain the supernatant; (3) adjust the pH of the supernatant to 3 with 10% hydrochloric acid, stir, stand, centrifuge, remove the precipitate, and obtain the supernatant; (4) pass the supernatant through a macroporous adsorption resin D4020, elute with dilute ethanol, and obtain the eluate; (5) reduce the pressure to concentrate the eluate, the vacuum degree is greater than 0.05 Mpa, vacuum freeze dry the extract, the vacuum degree is greater than 0.02 Mpa, and the drying time is 4 hours; and a total of 113 g of dry product is prepared.

[0031] The content of the pentacyclic triterpenoids and flavonoids in the dry product is 89% and 67% respectively, which is determined by UPLC. Example 4

[0032] (1) Take the roots of Actinidia 2Kg, Snowygrass 3Kg, Oldenlandia 2Kg, Centella asiatica 2Kg, and crush them into coarse powder. Add 10 times the amount of 90% ethanol and heat to reflux for 90 minutes. Filter, add 8 times the amount of 90% ethanol to the residue and heat to reflux for 60 minutes. Filter, add 6 times the amount of 90% ethanol to the residue and heat to reflux for 60 minutes. Combine the three filtrates, recover the ethanol by vacuum concentration, and continue to concentrate to a hot specific gravity of 1.05. (2) Add 5g of chitosan, stir, stand, centrifuge to remove the precipitate, and obtain the supernatant. (3) Adjust the pH of the supernatant obtained in step (2) to 2 with 10% hydrochloric acid, stand, centrifuge to remove the precipitate, and obtain the supernatant. (4) Pass the supernatant obtained in step (3) through a D4020 macroporous adsorption resin column, and elute with dilute ethanol solution to obtain the eluate. (5) Concentrate the eluate under reduced pressure, with a vacuum degree greater than 0.05Mpa, to obtain the extract, and freeze-dry the extract under vacuum, with a vacuum degree greater than 0.02Mpa, for 4 hours. A total of 125g of dry product is obtained.

[0033] The content of pentacyclic triterpenoids in the dry product is 90% and the content of flavonoids is 74% as determined by UPLC. Example 5

[0034] (1) Take the roots of Actinidia 2Kg, Snowygrass 3Kg, Oldenlandia 2Kg, Centella asiatica 2Kg, and crush them into coarse powder. Add 10 times the amount of 90% ethanol and heat to reflux for 90 minutes. Filter, add 8 times the amount of 90% ethanol to the residue and heat to reflux for 60 minutes. Filter, add 6 times the amount of 90% ethanol to the residue and heat to reflux for 60 minutes. Combine the three filtrates, recover the ethanol by vacuum concentration, and continue to concentrate to a hot specific gravity of 1.05. (2) Add 5g of chitosan, stir, stand, centrifuge to remove the precipitate, and obtain the supernatant. (3) Adjust the pH of the supernatant obtained in step (2) to 2 with 10% hydrochloric acid, stand, centrifuge to remove the precipitate, and obtain the supernatant. (4) Pass the supernatant obtained in step (3) through a D4020 macroporous adsorption resin column, and elute with dilute ethanol solution to obtain the eluate. (5) Concentrate the eluate under reduced pressure, with a vacuum degree greater than 0.05Mpa, to obtain the extract, and freeze-dry the extract under vacuum, with a vacuum degree greater than 0.02Mpa, for 4 hours. A total of 125g of dry product is obtained.

[0035] The content of pentacyclic triterpenoids in the dry product is 90% and the content of flavonoids is 74% as determined by UPLC. Example 6

[0036] (1) Take 2Kg of the root of Actinidia, 1Kg of Rabdosia rubescens, 2Kg of Oldenlandia, and 2Kg of Centella asiatica, crush them into coarse powder, add 10 times of 90% ethanol to heat and reflux, time 90 minutes; filter, add 8 times of 90% ethanol to the residue to heat and reflux, time 60 minutes; filter, add 6 times of 90% ethanol to the residue to heat and reflux, time 60 minutes, combine the three filtrates; vacuum concentration, recover ethanol, continue to concentrate to the hot specific gravity of 1.05; (2) add 5g of chitosan, stir, stand, centrifugal to remove the precipitate, get the supernatant; (3) adjust the pH of the supernatant obtained in step (2) to 2 with 10% hydrochloric acid, stand, centrifugal to remove the precipitate, get the supernatant; (4) pass the supernatant obtained in step (3) through a macroporous adsorption resin D4020 column, elute with dilute NaOH solution; (5) reduce pressure to concentrate the eluent obtained in step (4), vacuum degree greater than 0.05Mpa, get the extract, vacuum freeze dry the extract, vacuum degree greater than 0.02Mpa, drying time 4 hours; a total of 116g of dry product is prepared.

[0037] The content of pentacyclic triterpenoids in the dry product is quantitatively determined by UPLC to be 92%, and the content of flavonoids is 68%. Example 7

[0038] Take 1Kg of the crude drug of the application prepared in Example 2, add medicinal starch, dextrin, and 80% ethanol to prepare soft material, mix well, pass through a 12-mesh sieve, wet granulation, dry (below 60℃), and sieve (pass through a 14-mesh sieve) to prepare finished granules, which are packaged into bags to obtain the granules of the preparation of the application. Example 8

[0039] Take 1Kg of the crude drug of the application prepared in Example 1, add 2Kg of medicinal microcrystalline cellulose, mix well, wet granulation, adjust with water, and prepare granules that are loose and pass through a 20-mesh sieve, dry, and then fill into capsules to obtain the capsules of the preparation of the application. Example 9

[0040] Take 1Kg of the crude drug of the application prepared in Example 3, add 6Kg of PEG6000 (polyethylene glycol 6000), heat to 60℃ to melt, mix well, place in a drop pill machine, cool with liquid paraffin, and prepare drop pills to obtain the drop pills of the preparation of the application. Example 10

[0041] Take 1Kg of the crude drug of the application prepared in Example 4, add 2Kg of mannitol, dissolve with sterilized water, pass through a 0.3um microporous filter membrane, and then pour into an ampoule, freeze dry at -50℃ for 24 hours to obtain the freeze-dried powder injection of the preparation of the application. Example 11

[0042] Colon cancer HCT116, DIFI cells were cultured in DMEM medium containing 10% fetal bovine serum, and when the cells were logarithmically growing, they were inoculated in sterile 96-well culture plates at a density of 2X106 / ml, 100 ul of tumor cell suspension was added to each well, 10 ul of active ingredient extract solution obtained in Example 1 (concentration 1 mg / ml) was added, 6 wells per group, the same amount of PBS solution 10 ul was added to the normal control wells, and the same amount of compound smilax soup water extract 10 ul was added to the negative control wells, then they were shaken and mixed, and then they were placed in a 5% CO2 incubator (37°C) for 24 h, CCK-8 solution was added to each well, 10 ul, and incubation was continued for 2 h, and then the optical density OD value was measured at a wavelength of 450 nm using an enzyme-linked immunoassay analyzer (model: 3001, Thermo Fisher Scientific, USA), and the results were calculated.

[0043] Calculation formula:

[0044] Inhibition rate = 1 - (measured value - blank value) / (control value - blank value) x 100%.

[0045] In the colon cancer HCT116, DIFI cell inhibition rate experiment, it was found that the inhibition rate of the preparation of the application on the two tumor cells was significantly higher than that of the existing compound smilax three herb soup water extract (concentrated solution of filtrate obtained by adding 10 times the weight of boiling water for extraction for 2 hours), and the inhibition rate increased with the increase of the dose. The results are shown in the following table.

[0046] Example 12

[0047] Experimental method: Take mouse-derived colon cancer MC-38 cells, adjust the cell number to 5X105 / ml concentration, inject 0.2 ml into the right axillary subcutaneous tissue of Babl / C mice (4 weeks old, female), and after 2 weeks, randomly group the mice for administration, 5-fluorouracil (5-FU) group intraperitoneal administration, normal saline (control group), existing compound smilax three herb soup decoction group (concentrated solution of filtrate obtained by adding 10 times the weight of boiling water for extraction for 2 hours), and the preparation of the application of Example 1 (dose 10 g / Kg) group gavage, on the 28th day, weigh the mice, take the tumor mass, weigh the tumor and measure the tumor volume, and calculate the tumor inhibition rate.

[0048] The tumor volume calculation formula is: V = 1 / 2 x a x b2 (a is the long axis and b is the short axis);

[0049] Tumor inhibition rate (%) = (W[negative control group] - W[administration experiment group]) / W[negative control group] x 100%;

[0050] Results: The existing compound Zhenli Sancao decoction water decoction on mice tumor inhibition rate was 35.5%, and the same dose of the preparation of the present application tumor inhibition rate was 56.6~68.7%, compared with the control group has a significant difference; and compared with the 5-FU group, the difference is not significant; suggest that the preparation of the present application has good anti-tumor effect, and its tumor inhibition effect increases with the dose. The results are shown in the following table.

[0051] .

Claims

1. A compound preparation of effective active ingredients from traditional Chinese medicine for treating colorectal cancer, made from the following raw materials in parts by weight: 1-3 parts of *Actinidia chinensis* root, 1-3 parts of *Rhizoma Scutellariae*, 1-3 parts of *Hedyotis diffusa*, and 1-3 parts of *Centella asiatica*; characterized in that, The extraction method for effective active ingredients in compound traditional Chinese medicine raw materials such as *Actinidia chinensis* root, *Rhizoma Rabdosiae*, *Hedyotis diffusa*, and *Centella asiatica* includes the following steps: (1) Mix the raw medicinal materials of *Actinidia chinensis* root, *Rabdosia rubescens*, *Hedyotis diffusa*, and *Centella asiatica* or their pulverized parts, and extract them with 90% ethanol for 1-2 hours. Filter, centrifuge to remove the precipitate, and retain the extract. Repeat the extraction 2-3 times and combine the extracts. (2) Add chitosan to the extract obtained in step (1), stir, let it stand to precipitate, filter or centrifuge to remove the precipitate, and obtain filtrate or supernatant. (3) Add pH adjuster to the filtrate or supernatant obtained in step (2) to adjust the pH value, let it stand to precipitate, filter or centrifuge to remove the precipitate, and obtain filtrate or supernatant. (4) Elute the filtrate or supernatant obtained in step (3) onto macroporous adsorption resin D4020 to obtain an eluent; (5) The eluent obtained in step (4) is concentrated under reduced pressure to obtain an extract. The extract is dried and pulverized into a fine powder to obtain the original drug of the effective active ingredient preparation of the traditional Chinese medicine compound for anti-colorectal cancer of the present invention.

2. The herbal compound preparation for treating colorectal cancer according to claim 1, characterized in that, In step (1), the extraction with 90% ethanol is carried out. The mass ratio of the Chinese herbal medicine raw material to 90% ethanol is: 1:10 for the first time, 1:8 for the second time, and 1:6 for the third time.

3. The herbal compound preparation for treating colorectal cancer according to claim 1 or 2, characterized in that, In step (2), the amount of chitosan added is 0.1 to 1% of the weight of the Chinese herbal medicine raw material.

4. The herbal compound preparation for treating colorectal cancer according to claim 1 or 2, characterized in that, In step (3), the pH value is adjusted to 2-5.

5. The herbal compound preparation for treating colorectal cancer according to claim 3, characterized in that, In step (3), the pH value is adjusted to 2-5.

6. The herbal compound preparation of effective active ingredients for treating colorectal cancer according to claim 1 or 2, characterized in that, The pH adjuster is hydrochloric acid or acetic acid.

7. The herbal compound preparation for treating colorectal cancer according to claim 3, characterized in that, The pH adjuster is hydrochloric acid or acetic acid.

8. The herbal compound preparation for treating colorectal cancer according to claim 4, characterized in that, The pH adjuster is hydrochloric acid or acetic acid.

9. The herbal compound preparation for treating colorectal cancer according to claim 1 or 2, characterized in that, In step (5), the drying process is vacuum drying, spray drying, or freeze drying.

10. The herbal compound preparation for treating colorectal cancer according to claim 3, characterized in that, In step (5), the drying process is vacuum drying, spray drying, or freeze drying.

11. The herbal compound preparation for treating colorectal cancer according to claim 4, characterized in that, In step (5), the drying process is vacuum drying, spray drying, or freeze drying.

12. The herbal compound preparation for treating colorectal cancer according to claim 6, characterized in that, In step (5), the drying process is vacuum drying, spray drying, or freeze drying.

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