Traditional Chinese medicine composition for treating tumors and preparation method thereof

Abstract

The invention relates to the technical field of traditional Chinese medicine compositions, in particular to a traditional Chinese medicine composition for treating tumors and a preparation method thereof. The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 3 to 6 parts of traditional Chinese medicine extract, 5 to 10 parts of modified SiO2 and 8 to 12 parts of cellulose-chitosan compound, the traditional Chinese medicine extract is prepared from the following raw material components: 20 to 30 parts of herba scutellariae barbatae, 20 to 30 parts of herba hedyotidis diffusae, 15 to 25 parts of radix astragali seu hedysari, 10 to 20 parts of radix codonopsis, 10 to 15 parts of radix notoginseng and 5 to 10 parts of radix glycyrrhizae. The prepared traditional Chinese medicine composition can effectively inhibit tumor growth and has high biological safety and bioavailability.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine compositions, and particularly relates to a traditional Chinese medicine composition for treating tumors and a preparation method thereof. Background Art

[0002] Tumors are major diseases threatening human health. Existing treatment methods mainly include surgery, radiotherapy, chemotherapy, targeted therapy, etc. However, these methods all have certain limitations. For example, surgery has large trauma, obvious side effects of radiotherapy and chemotherapy, high cost of targeted therapy and limitations in applicable populations, etc. Traditional Chinese medicine has unique advantages in tumor treatment. It can regulate the physiological functions of the human body as a whole, enhance the body's immunity, reduce the adverse reactions of radiotherapy and chemotherapy, and inhibit the growth and metastasis of tumor cells. However, existing traditional Chinese medicine drugs for treating tumors still have room for improvement in terms of efficacy, safety and stability. Therefore, developing a highly effective and safe traditional Chinese medicine composition for treating tumors is a technical problem to be solved. Summary of the Invention

[0003] In view of the deficiencies of the prior art, the present invention provides a traditional Chinese medicine composition for treating tumors and a preparation method thereof.

[0004] The present invention is achieved through the following technical solutions: A traditional Chinese medicine composition for treating tumors, the preparation raw materials of which include the following components in parts by weight: 3 - 6 parts of traditional Chinese medicine extract, 5 - 10 parts of modified SiO₂, and 8 - 12 parts of cellulose-chitosan complex.

[0005] Further, the preparation raw materials of the traditional Chinese medicine extract include the following components in parts by weight: 20 - 30 parts of Scutellaria barbata, 20 - 30 parts of Hedyotis diffusa, 15 - 25 parts of Astragalus membranaceus, 10 - 20 parts of Codonopsis pilosula, 10 - 15 parts of Panax notoginseng, and 5 - 10 parts of Glycyrrhiza uralensis.

[0006] Further, the preparation method of the traditional Chinese medicine extract includes the following steps: taking each medicinal material, drying and pulverizing it through an 80 - 100 mesh sieve, adding it to 60 - 70 vol% ethanol according to a material-liquid ratio of 1 g:20 - 30 mL, soaking for 30 min, heating and refluxing for 2 h, filtering, extracting 3 times, combining the filtrates, rotary evaporating to 20% of the original volume, and freeze-drying to obtain the traditional Chinese medicine extract.

[0007] Further, the preparation method of the modified SiO₂ includes the following steps: A1: Disperse CTAB in deionized water, add 2 M NaOH solution, stir at 80 °C and 600 - 800 r / min for 30 min, dropwise add TEOS at a rate of 0.2 mL / min, continue stirring for 4 - 6 h, cool to room temperature, filter, wash the precipitate with ethanol and deionized water, dry in vacuum, and calcine in a muffle furnace at 550 °C for 5 - 6 h to obtain mesoporous SiO₂; A2: Add the mesoporous SiO₂ obtained in step A1 into ethanol, ultrasonicate at 100 - 150 W for 30 min, add APTES, heat under reflux for 12 h, centrifuge at 10000 r / min for 5 - 7 min, wash the precipitate with ethanol and water, dry in vacuum to obtain amino-functionalized mesoporous SiO₂; A3: Add the amino-functionalized mesoporous SiO₂ obtained in step A2 into DMF, ultrasonicate at 100 - 150 W for 30 min, add succinic anhydride, stir at 600 - 800 rpm for 24 h, centrifuge at 10000 rpm for 10 min, wash the precipitate with water and ethanol, dry in vacuum to obtain modified SiO₂.

[0008] Further, in step A1, the mass concentration of CTAB in deionized water is 1.5 - 2 mg / mL.

[0009] Further, in step A1, the dosage ratio of CTAB to NaOH solution is 2 g:7 mL.

[0010] Further, in step A1, the dosage ratio of CTAB to TEOS is 1 g:5 mL.

[0011] Further, in step A2, the mass concentration of mesoporous SiO₂ in ethanol is 5 - 10 mg / mL.

[0012] Further, in step A2, the dosage ratio of mesoporous SiO₂ to APTES is 1 g:1 - 1.5 mL.

[0013] Further, in step A3, the mass concentration of amino-functionalized mesoporous SiO₂ in DMF is 8 - 12 mg / mL.

[0014] Further, in step A3, the mass ratio of amino-functionalized mesoporous SiO₂ to succinic anhydride is 1:4 - 6.

[0015] Further, the raw materials for preparing the cellulose-chitosan composite include the following components in parts by weight: chitosan (CS) 4 - 6 parts, cellulose 2 - 3 parts, thionyl chloride 5 - 8 parts, 1,3-propanediamine 12 - 18 parts, 2,2'-dipyridyl disulfide 1 - 2 parts.

[0016] Further, the preparation method of the cellulose-chitosan composite comprises the following steps: B1: Weigh NAC and add it to DMF. After stirring and dissolving, add EDC·HCl and NHS, and react in the dark at room temperature for 12 h to obtain an activation solution. Weigh chitosan and dissolve it in 0.1 M HCl, stir for 5 h, add the activation solution, adjust the pH of the solution to 5-6 with 1 M NaOH solution, react in the dark at room temperature for 24 h, dialyze, and freeze-dry to obtain modified chitosan; B2: Add cellulose to DMF and stir evenly. Dropwise add thionyl chloride under stirring at 80-90 °C. After the addition is complete, continue to react at 80-90 °C for 2-3 h. Cool to room temperature, add 10 times the volume of cold water while stirring, filter, vacuum-dry the precipitate, grind it into powder, add it to distilled water, add 1,3-propanediamine while stirring, heat and reflux for 3-4 h, filter, wash the precipitate with deionized water, and then vacuum-dry; B3: Prepare the activation solution using NAC according to the method in step B1. Add the product obtained after vacuum drying in step B2 to DMF, stir at 200-300 rpm for 1 h, add the activation solution, stir and react at room temperature for 5-6 h, dialyze with 5 mM HCl solution for 48 h, dialyze with ultrapure water for 24 h, and freeze-dry to obtain modified cellulose; B4: Add the modified chitosan obtained in step B1 to ultrapure water and mix evenly. Add 2,2'-dipyridyl disulfide to methanol and mix evenly. Mix the two, react at room temperature for 24 h, centrifuge at 8000 rpm for 10-15 min, wash with methanol and deionized water, dry, disperse it with the modified cellulose obtained in step B3 in DMF, react at 30 °C for 24 h, centrifuge at 8000 rpm for 10-15 min, wash with DMF and deionized water, and then dry to obtain the cellulose-chitosan composite.

[0017] Further, in step B1, the mass ratio of chitosan to NAC is 2:1.

[0018] Further, in step B1, the mass concentration of NAC in DMF is 20-30 mg / mL.

[0019] Further, in step B1, the mass ratio of NAC, EDC·HCl and NHS is 2:5:3.

[0020] Further, in step B1, the mass-volume fraction of chitosan in 0.1 M HCl is 1 w / v%.

[0021] Further, in step B1, the dialysis is carried out as follows: under the condition of avoiding light at 4 °C, in a dialysis bag with a molecular weight cut-off of 3500 Da, dialysis is carried out with 5 mM HCl solution for 24 h, with 5 mM HCl solution containing 1 w / v% NaCl for 24 h, with 1 mM HCl solution for 24 h, and with ultrapure water for 12 h.

[0022] Further, in step B2, the mass concentration of the cellulose in DMF is 40 - 50 mg / mL.

[0023] Further, in step B2, the dosage ratio of the cellulose to distilled water is 1 g:40 - 50 mL.

[0024] Further, in step B3, the mass ratio of the NAC to the cellulose is 1:2.

[0025] Further, in step B3, the dosage ratio of the DMF to the cellulose is 100 mL:1 - 2 g.

[0026] Further, in step B4, the mass concentration of the 2,2'-dipyridyl disulfide in methanol is 20 - 25 mg / mL.

[0027] Further, in step B4, the mass concentration of the modified chitosan in ultrapure water is 10 mg / mL.

[0028] Further, in step B4, the mass concentration of the modified cellulose in DMF is 10 mg / mL.

[0029] Further, the present invention also provides a preparation method of the traditional Chinese medicine composition for treating tumors, comprising the following steps: S1: Dispersing the modified SiO2 and the traditional Chinese medicine extract in ethanol, stirring at 37 °C and 300 - 400 rpm for 24 h, centrifuging at 10000 rpm for 10 min, washing the precipitate with ethanol and drying it under vacuum to obtain the drug-loaded modified SiO2; S2: Adding the drug-loaded modified SiO2, EDC·HCl and NHS obtained in step S1 into a PBS buffer solution with pH 5, stirring at room temperature for 1 h, centrifuging at 10000 rpm for 10 min, and collecting the precipitate; S3: Dispersing the precipitate obtained in step S2 in a PBS buffer solution with pH 7.4, adding a cellulose-chitosan complex, stirring and reacting at room temperature for 12 h, centrifuging at 10000 rpm for 10 min, washing the precipitate with water and ethanol, and drying it under vacuum to obtain the traditional Chinese medicine composition for treating tumors.

[0030] Further, in step S1, the mass concentration of the modified SiO2 in ethanol is 5 mg / mL.

[0031] Further, in step S2, the mass ratio of the modified SiO2, EDC, and NHS is 1:1.5:1.

[0032] Further, in step S2, the mass concentration of EDC·HCl in PBS at pH 5 is 15 mg / mL.

[0033] Further, in step S3, the volume ratio of the PBS buffer solution at pH 7.4 to the PBS buffer solution at pH 5 is 1:1.

[0034] Compared with the prior art, the present invention has the following beneficial effects: The present invention provides a traditional Chinese medicine composition for treating tumors. By scientifically combining traditional Chinese medicine extracts with modified SiO2 and cellulose-chitosan composites, the modified SiO2 has good drug-loading performance, and the cellulose-chitosan composite can protect and modify the drug-loading system, and through their synergistic effect, the therapeutic effect on tumors is enhanced. In the present invention, traditional Chinese medicine materials such as Scutellaria barbata, Hedyotis diffusa, Astragalus membranaceus, Codonopsis pilosula, Panax notoginseng, and Glycyrrhiza uralensis are selected to prepare extracts. Among them, Scutellaria barbata and Hedyotis diffusa have the effects of clearing heat and detoxifying, dissipating swelling and anti-cancer. Modern research shows that the various active ingredients contained in them can induce apoptosis of tumor cells and inhibit the formation of tumor blood vessels; Astragalus membranaceus and Codonopsis pilosula can invigorate qi and strengthen the spleen, enhance the body's immunity, and by regulating the body's immune function, activate immune cells, and enhance the recognition and killing ability of tumor cells; Panax notoginseng has the effects of promoting blood circulation to remove blood stasis, dissipating swelling and relieving pain, can improve the microcirculation of tumor tissues, and inhibit the invasion and metastasis of tumor cells; Glycyrrhiza uralensis can coordinate the actions of various drugs, and at the same time has certain immune regulation and anti-inflammatory effects. These traditional Chinese medicine materials are combined with each other to play a role in inhibiting tumor growth and metastasis from multiple pathways and multiple targets. In the present invention, mesoporous SiO2 is prepared, which can load traditional Chinese medicine extracts. It is aminated by APTES to introduce amino groups, providing active sites for subsequent modification. Then it reacts with succinic anhydride, and carboxyl groups are introduced onto the surface of the carrier through acylation reaction, so that carboxyl groups are connected to the surface of SiO2, providing sites for subsequent reactions and enhancing the interaction with cellulose-chitosan; the modified SiO2 is used as a drug carrier to effectively load traditional Chinese medicine extracts, realizing the slow release of drugs, prolonging the action time of drugs in the body, improving the bioavailability of drugs, and at the same time reducing the toxic and side effects of drugs. In the present invention, chitosan-cellulose composites are prepared. Both chitosan and cellulose are natural polymer materials with good biocompatibility. By chemically modifying chitosan and cellulose and introducing active groups, the two can be combined through chemical bonds to form a composite. This composite can further encapsulate the drug-loaded modified SiO2 to form a stable drug delivery system, enhancing the drug slow-release effect and improving the bioavailability. In the present invention, after NAC is activated, it reacts with some amino groups of chitosan to introduce mercapto groups onto chitosan, which is beneficial to subsequent cross-linking reactions; cellulose hydroxyl groups are activated with thionyl chloride and then reacted with 1,3-propanediamine to introduce amino groups, which is beneficial to increasing the amino content and enhancing the interaction with the carboxyl group-containing drug-loaded SiO2. Then NAC is used to introduce mercapto groups onto cellulose. Using 2,2'-dipyridyl disulfide as a cross-linking agent, through the mercapto-disulfide exchange reaction, the modified chitosan and the modified cellulose are connected by disulfide bonds (-S-S-), reducing self-cross-linking reactions. The formed cellulose-chitosan composite has a special molecular structure and physical and chemical properties, and can encapsulate and protect the drug-loaded modified SiO2.During the drug release process, this system can effectively slow down the drug release rate, prevent premature drug release, thereby prolonging the action time of the drug in the body and improving the utilization efficiency of the drug. In addition, it can reduce the potential biological toxicity of the drug and improve biocompatibility, and can be used as a safe oral drug delivery carrier. The traditional Chinese medicine composition of the present invention improves the absorption process of the drug in the body through reasonable ingredient combination and drug delivery system design. The drug delivery system formed by modified SiO2 and cellulose-chitosan complex can protect the drug from premature degradation in the gastrointestinal environment and promote drug absorption. It has a better absorption effect in the body, significantly improves the oral bioavailability, is beneficial to enhancing the therapeutic potential of the drug, and reducing the dosage and frequency of medication. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. Obviously, the drawings in the following description are only those of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0036] Figure 1 For the tumor treatment effect of the traditional Chinese medicine composition described in Example 1 of the present invention; Figure 2 For the biological safety of the traditional Chinese medicine compositions described in Example 2 of the present invention and Comparative Examples 1-3; Figure 3 For the sustained-release effect of the traditional Chinese medicine compositions described in Example 3 of the present invention and Comparative Examples 1-3; Figure 4 For the bioavailability of the traditional Chinese medicine compositions described in Example 1 of the present invention and Comparative Examples 1-3. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0037] To make the objectives, technical solutions, and advantages of the present invention clearer, the following further details the present invention with reference to specific embodiments. However, the present invention is not limited to the following embodiments. It should be noted that, unless otherwise specified, the chemical reagents involved in the present invention are purchased through commercial channels.

[0038] Example 1: A traditional Chinese medicine composition for treating tumors, the preparation raw materials include the following components in parts by weight: 6 parts of traditional Chinese medicine extract, 10 parts of modified SiO2, and 12 parts of cellulose-chitosan complex.

[0039] The preparation raw materials of the traditional Chinese medicine extract include the following components in parts by weight: 30 parts of Scutellaria barbata, 30 parts of Hedyotis diffusa, 25 parts of Astragalus membranaceus, 20 parts of Codonopsis pilosula, 15 parts of Panax notoginseng, and 10 parts of Glycyrrhiza uralensis.

[0040] Preparation method of traditional Chinese medicine extract, comprising the following steps: drying and pulverizing each medicinal material, passing through a 100-mesh sieve, adding it to 70 vol% ethanol according to a material-liquid ratio of 1 g: 30 mL, soaking for 30 min, heating under reflux for extraction for 2 h, filtering, extracting 3 times, combining the filtrates, rotary evaporating to 20% of the original volume, and freeze-drying to obtain the traditional Chinese medicine extract.

[0041] Preparation method of modified SiO2, comprising the following steps: A1: Dissolve 2 g of CTAB in 1000 mL of deionized water, add 7 mL of 2 M NaOH solution, stir at 80 °C and 800 r / min for 30 min, dropwise add 10 mL of TEOS at a rate of 0.2 mL / min, continue stirring for 6 h, cool to room temperature, filter, wash the precipitate with ethanol and deionized water, dry under vacuum, and calcine in a muffle furnace at 550 °C for 6 h to obtain mesoporous SiO2; A2: Add 2 g of the mesoporous SiO2 obtained in step A1 to 200 mL of ethanol, ultrasonicate at 150 W for 30 min, add 3 mL of APTES, heat under reflux for 12 h, centrifuge at 10000 r / min for 7 min, wash the precipitate with ethanol and water, dry under vacuum to obtain amino-functionalized mesoporous SiO2; A3: Add 1.2 g of the amino-functionalized mesoporous SiO2 obtained in step A2 to 100 mL of DMF, ultrasonicate at 150 W for 30 min, add 7.2 g of succinic anhydride, stir at 800 rpm for 24 h, centrifuge at 10000 rpm for 10 min, wash the precipitate with water and ethanol, dry under vacuum to obtain modified SiO2.

[0042] Raw materials for preparing cellulose-chitosan composite, comprising the following components in parts by weight: 6 parts of chitosan, 3 parts of cellulose, 8 parts of thionyl chloride, 18 parts of 1,3-propanediamine, and 2 parts of 2,2'-dipyridyldisulfide.

[0043] Preparation method of cellulose-chitosan composite, comprising the following steps: B1: Weigh 3 g of NAC and add it to 100 mL of DMF. After stirring and dissolving, add 7.5 g of EDC·HCl and 4.5 g of NHS, and react for 12 h in the dark at room temperature to obtain an activation solution. Weigh 6 g of chitosan and dissolve it in 600 mL of 0.1 M HCl, stir for 5 h, add the activation solution, adjust the pH of the solution to 6 with 1 M NaOH solution, react for 24 h in the dark at room temperature, dialyze, and freeze-dry to obtain modified chitosan; the dialysis is carried out under light-shielded conditions at 4°C, in a dialysis bag with a molecular weight cut-off of 3500 Da, dialyze with 5 mM HCl solution for 24 h, dialyze with 5 mM HCl solution containing 1 w / v% NaCl for 24 h, dialyze with 1 mM HCl solution for 24 h, and dialyze with ultrapure water for 12 h; B2: Add 3 g of cellulose to 60 mL of DMF and stir well. Dropwise add 8 g of thionyl chloride while stirring at 90°C. After the addition is complete, continue to react at 90°C for 3 h. Cool to room temperature, add 10 times the volume of cold water while stirring, filter, vacuum-dry the precipitate, grind it into powder, add it to 150 mL of distilled water, add 18 g of 1,3-propanediamine while stirring, heat under reflux for 4 h, filter, wash the precipitate with deionized water, and then vacuum-dry; B3: Prepare an activation solution using 1.5 g of NAC according to the method of step B1. Add the product obtained after vacuum-drying in step B2 to 150 mL of DMF, stir at 300 rpm for 1 h, add the activation solution, stir and react at room temperature for 6 h, dialyze with 5 mM HCl solution for 48 h, dialyze with ultrapure water for 24 h, and freeze-dry to obtain modified cellulose; B4: Add the modified chitosan obtained in step B1 to ultrapure water and mix well. The mass concentration of the modified chitosan in ultrapure water is 10 mg / mL. Add 2 g of 2,2'-dipyridyl disulfide to 80 mL of methanol and mix well. Mix the two, react at room temperature for 24 h, centrifuge at 8000 rpm for 15 min, wash with methanol and deionized water, dry, and disperse it with the modified cellulose obtained in step B3 in DMF. The mass concentration of the modified cellulose in DMF is 10 mg / mL. React at 30°C for 24 h, centrifuge at 8000 rpm for 15 min, wash with DMF and deionized water, and then dry to obtain a cellulose-chitosan composite.

[0044] This example also provides a preparation method of the traditional Chinese medicine composition for treating tumors, including the following steps: S1: Disperse 10 g of modified SiO2 and 6 g of traditional Chinese medicine extract in 2000 mL of ethanol, stir at 37°C and 400 rpm for 24 h, centrifuge at 10000 rpm for 10 min, wash the precipitate with ethanol and vacuum-dry to obtain drug-loaded modified SiO2; S2: Add the drug-loaded modified SiO₂, EDC·HCl, and NHS obtained in step S1 into a PBS buffer solution with a pH of 5. The mass ratio of modified SiO₂, EDC, and NHS is 1:1.5:1. The mass concentration of EDC·HCl in PBS with a pH of 5 is 15 mg / mL. Stir at room temperature for 1 h, centrifuge at 10000 rpm for 10 min, and collect the precipitate. S3: Disperse the precipitate obtained in step S2 in a PBS buffer solution with a pH of 7.4. The volume ratio of the PBS buffer solution with a pH of 7.4 to the PBS buffer solution with a pH of 5 is 1:1. Add 12 g of cellulose-chitosan complex, stir and react at room temperature for 12 h, centrifuge at 10000 rpm for 10 min, wash the precipitate with water and ethanol, and dry it under vacuum to obtain a traditional Chinese medicine composition for treating tumors.

[0045] Example 2: A traditional Chinese medicine composition for treating tumors, the preparation raw materials of which include the following components in parts by weight: 3 parts of traditional Chinese medicine extract, 5 parts of modified SiO₂, and 8 parts of cellulose-chitosan complex.

[0046] The preparation raw materials of the traditional Chinese medicine extract include the following components in parts by weight: 20 parts of Scutellaria barbata, 20 parts of Hedyotis diffusa, 15 parts of Astragalus membranaceus, 10 parts of Codonopsis pilosula, 10 parts of Panax notoginseng, and 5 parts of Glycyrrhiza uralensis.

[0047] The preparation method of the traditional Chinese medicine extract includes the following steps: Take each medicinal material, dry it, crush it, and pass it through an 80-mesh sieve. Add it to 60 vol% ethanol according to a material-liquid ratio of 1 g:20 mL, soak for 30 min, heat under reflux for 2 h, filter, extract 3 times, combine the filtrates, rotary evaporate to 20% of the original volume, and freeze-dry to obtain the traditional Chinese medicine extract.

[0048] The preparation method of modified SiO₂ includes the following steps: A1: Take 3 g of CTAB and disperse it in 2000 mL of deionized water. Add 10.5 mL of 2 M NaOH solution, stir at 80 °C and 600 r / min for 30 min, dropwise add 15 mL of TEOS at a rate of 0.2 mL / min, continue to stir for 4 h, cool to room temperature, filter, wash the precipitate with ethanol and deionized water, dry it under vacuum, and calcine it in a muffle furnace at 550 °C for 5 h to obtain mesoporous SiO₂. A2: Add 2 g of the mesoporous SiO₂ obtained in step A1 into 400 mL of ethanol, ultrasonicate at 100 W for 30 min, add 2 mL of APTES, heat under reflux for 12 h, centrifuge at 10000 r / min for 5 min, wash the precipitate with ethanol and water, and dry it under vacuum to obtain amino-functionalized mesoporous SiO₂. A3: Add 0.8 g of the amidated mesoporous SiO₂ obtained in step A2 into 100 mL of DMF, ultrasonicate for 30 min at 100 W, add 3.2 g of succinic anhydride, stir at 600 rpm for 24 h, centrifuge at 10000 rpm for 10 min, wash the precipitate with water and ethanol, and dry it under vacuum to obtain modified SiO₂.

[0049] The raw materials for preparing the cellulose-chitosan composite include the following components in parts by weight: 4 parts of chitosan, 2 parts of cellulose, 5 parts of thionyl chloride, 12 parts of 1,3-propanediamine, and 1 part of 2,2'-dipyridyl disulfide.

[0050] The preparation method of the cellulose-chitosan composite includes the following steps: B1: Weigh 2 g of NAC and add it into 100 mL of DMF. After stirring and dissolving, add 5 g of EDC·HCl and 3 g of NHS, and react in the dark at room temperature for 12 h to obtain an activation solution. Weigh 4 g of chitosan and dissolve it in 400 mL of 0.1 M HCl, stir for 5 h, add the activation solution, adjust the pH of the solution to 5 with 1 M NaOH solution, react in the dark at room temperature for 24 h, dialyze, and freeze-dry to obtain modified chitosan; dialysis is carried out under dark conditions at 4°C in a dialysis bag with a molecular weight cut-off of 3500 Da, dialyze with 5 mM HCl solution for 24 h, dialyze with 5 mM HCl solution containing 1 w / v% NaCl for 24 h, dialyze with 1 mM HCl solution for 24 h, and dialyze with ultrapure water for 12 h; B2: Add 2 g of cellulose into 50 mL of DMF and stir evenly. Dropwise add 5 g of thionyl chloride while stirring at 80°C. After the addition is complete, continue to react at 80°C for 2 h. Cool to room temperature, add 10 times the volume of cold water while stirring, filter, vacuum-dry the precipitate, grind it into powder, add it into 80 mL of distilled water, add 12 g of 1,3-propanediamine while stirring, heat and reflux for 3 h, filter, wash the precipitate with deionized water, and then vacuum-dry; B3: Prepare an activation solution using 1 g of NAC according to the method in step B1. Add the product obtained after vacuum drying in step B2 into 200 mL of DMF, stir at 200 rpm for 1 h, add the activation solution, stir and react at room temperature for 5 h, dialyze with 5 mM HCl solution for 48 h, dialyze with ultrapure water for 24 h, and freeze-dry to obtain modified cellulose; B4: Add the modified chitosan obtained in step B1 to ultrapure water and mix well. The mass concentration of the modified chitosan in ultrapure water is 10 mg / mL. Add 1 g of 2,2'-dipyridyl disulfide to 50 mL of methanol and mix well. Mix the two, react at room temperature for 24 h, centrifuge at 8000 rpm for 10 min, wash with methanol and deionized water, dry, and disperse it with the modified cellulose obtained in step B3 in DMF. The mass concentration of the modified cellulose in DMF is 10 mg / mL. React at 30 °C for 24 h, centrifuge at 8000 rpm for 10 min, wash with DMF and deionized water and then dry to obtain the cellulose-chitosan composite.

[0051] This example also provides a preparation method of the traditional Chinese medicine composition for treating tumors, including the following steps: S1: Disperse 5 g of modified SiO2 and 3 g of traditional Chinese medicine extract in 1000 mL of ethanol, stir at 37 °C and 300 rpm for 24 h, centrifuge at 10000 rpm for 10 min, wash the precipitate with ethanol and vacuum dry to obtain the drug-loaded modified SiO2; S2: Add the drug-loaded modified SiO2, EDC·HCl and NHS obtained in step S1 to the PBS buffer solution with pH 5. The mass ratio of modified SiO2, EDC and NHS is 1:1.5:1. The mass concentration of EDC·HCl in the PBS with pH 5 is 15 mg / mL. Stir at room temperature for 1 h, centrifuge at 10000 rpm for 10 min, and collect the precipitate; S3: Disperse the precipitate obtained in step S2 in the PBS buffer solution with pH 7.4. The volume ratio of the PBS buffer solution with pH 7.4 to the PBS buffer solution with pH 5 is 1:1. Add 8 g of the cellulose-chitosan composite, stir and react at room temperature for 12 h, centrifuge at 10000 rpm for 10 min, wash the precipitate with water and ethanol, and vacuum dry to obtain the traditional Chinese medicine composition for treating tumors.

[0052] Example 3: A traditional Chinese medicine composition for treating tumors, the preparation raw materials of which include the following components in parts by weight: 5 parts of traditional Chinese medicine extract, 8 parts of modified SiO2, and 10 parts of cellulose-chitosan composite.

[0053] The preparation raw materials of the traditional Chinese medicine extract include the following components in parts by weight: 25 parts of Scutellaria barbata, 25 parts of Hedyotis diffusa, 20 parts of Astragalus membranaceus, 15 parts of Codonopsis pilosula, 12 parts of Panax notoginseng, and 7 parts of Glycyrrhiza uralensis.

[0054] Preparation method of traditional Chinese medicine extract, comprising the following steps: taking each medicinal material, drying, pulverizing, passing through a 90-mesh sieve, adding to 65 vol% ethanol according to a material-liquid ratio of 1 g:25 mL, soaking for 30 min, heating under reflux for extraction for 2 h, filtering, extracting 3 times, combining the filtrates, rotary evaporating to 20% of the original volume, and freeze-drying to obtain the traditional Chinese medicine extract.

[0055] Preparation method of modified SiO2, comprising the following steps: A1: Dissolve 2 g of CTAB in 1200 mL of deionized water, add 7 mL of 2 M NaOH solution, stir at 80 °C and 700 r / min for 30 min, dropwise add 10 mL of TEOS at a rate of 0.2 mL / min, continue stirring for 5 h, cool to room temperature, filter, wash the precipitate with ethanol and deionized water, dry in vacuum, and calcine in a muffle furnace at 550 °C for 5.5 h to obtain mesoporous SiO2; A2: Add 2 g of the mesoporous SiO2 obtained in step A1 to 300 mL of ethanol, ultrasonicate at 120 W for 30 min, add 2.5 mL of APTES, heat under reflux for 12 h, centrifuge at 10000 r / min for 6 min, wash the precipitate with ethanol and water, dry in vacuum to obtain amino-functionalized mesoporous SiO2; A3: Add 1 g of the amino-functionalized mesoporous SiO2 obtained in step A2 to 100 mL of DMF, ultrasonicate at 120 W for 30 min, add 5 g of succinic anhydride, stir at 700 rpm for 24 h, centrifuge at 10000 rpm for 10 min, wash the precipitate with water and ethanol, dry in vacuum to obtain modified SiO2.

[0056] Raw materials for preparing cellulose-chitosan composite, comprising the following components in parts by weight: 5 parts of chitosan, 2.5 parts of cellulose, 6 parts of thionyl chloride, 15 parts of 1,3-propanediamine, and 1.5 parts of 2,2'-dipyridyl disulfide.

[0057] Preparation method of cellulose-chitosan composite, comprising the following steps: B1: Weigh 2.5 g of NAC and add it to 100 mL of DMF. After stirring and dissolving, add 6.25 of EDC·HCl and 3.75 of NHS. React under dark at room temperature for 12 h to obtain an activation solution. Weigh 5 g of chitosan and dissolve it in 500 mL of 0.1 M HCl. Stir for 5 h, add the activation solution, adjust the pH of the solution to 5.5 with 1 M NaOH solution, react under dark at room temperature for 24 h, dialyze, and freeze-dry to obtain modified chitosan; Dialysis is carried out under dark at 4 °C in a dialysis bag with a molecular weight cut-off of 3500 Da, dialyze with 5 mM HCl solution for 24 h, dialyze with 5 mM HCl solution containing 1 w / v% NaCl for 24 h, dialyze with 1 mM HCl solution for 24 h, and dialyze with ultrapure water for 12 h; B2: Add 2.5 g of cellulose to 60 mL of DMF and stir well. Dropwise add 6 g of thionyl chloride under stirring at 85 °C. After the addition is complete, continue to react at 85 °C for 2.5 h. Cool to room temperature, add 10 times the volume of cold water while stirring, filter, vacuum-dry the precipitate, grind it into powder, add it to 120 mL of distilled water, add 15 g of 1,3-propanediamine under stirring, heat under reflux for 3.5 h, filter, wash the precipitate with deionized water and then vacuum-dry; B3: Prepare an activation solution using 1.25 g of NAC according to the method in step B1. Add the product obtained after vacuum-drying in step B2 to 200 mL of DMF, stir at 250 rpm for 1 h, add the activation solution, stir and react at room temperature for 5.5 h, dialyze with 5 mM HCl solution for 48 h, dialyze with ultrapure water for 24 h, and freeze-dry to obtain modified cellulose; B4: Add the modified chitosan obtained in step B1 to ultrapure water and mix well. The mass concentration of the modified chitosan in ultrapure water is 10 mg / mL. Add 1.5 g of 2,2'-dipyridyl disulfide to 70 mL of methanol and mix well. Mix the two, react at room temperature for 24 h, centrifuge at 8000 rpm for 12 min, wash with methanol and deionized water, dry, and disperse it with the modified cellulose obtained in step B3 in DMF. The mass concentration of the modified cellulose in DMF is 10 mg / mL. React at 30 °C for 24 h, centrifuge at 8000 rpm for 12 min, wash with DMF and deionized water and then dry to obtain a cellulose-chitosan complex.

[0058] This example also provides a preparation method of the traditional Chinese medicine composition for treating tumors, including the following steps: S1: Disperse 8 g of modified SiO2 and 5 g of traditional Chinese medicine extract in 1600 mL of ethanol, stir at 37 °C and 350 rpm for 24 h, centrifuge at 10000 rpm for 10 min, wash the precipitate with ethanol and vacuum-dry to obtain drug-loaded modified SiO2; S2: Add the drug-loaded modified SiO2, EDC·HCl, and NHS obtained in step S1 into a PBS buffer solution with a pH of 5. The mass ratio of modified SiO2, EDC, and NHS is 1:1.5:1. The mass concentration of EDC·HCl in PBS with a pH of 5 is 15 mg / mL. Stir at room temperature for 1 h, centrifuge at 10000 rpm for 10 min, and collect the precipitate. S3: Disperse the precipitate obtained in step S2 in a PBS buffer solution with a pH of 7.4. The volume ratio of the PBS buffer solution with a pH of 7.4 to the PBS buffer solution with a pH of 5 is 1:1. Add 10 g of cellulose-chitosan complex, stir and react at room temperature for 12 h, centrifuge at 10000 rpm for 10 min, wash the precipitate with water and ethanol, and dry it under vacuum to obtain a traditional Chinese medicine composition for treating tumors.

[0059] The difference between Comparative Example 1 and Example 1 is only that the cellulose-chitosan complex is not added, that is, steps S2 and S3 are not carried out.

[0060] The difference between Comparative Example 2 and Example 1 is only that mesoporous SiO2 is used to replace the modified SiO2.

[0061] The difference between Comparative Example 3 and Example 1 is only that chitosan is used to replace the modified chitosan.

[0062] Experimental Example 1: Take HepG2 cells in the logarithmic growth phase, digest them with trypsin, wash them with serum-free medium, count them, and prepare a single-cell suspension of 10×10 7 cells / mL. Subcutaneously inject 100 μL of the HepG2 cell suspension (i.e., 1×10 7 cells per mouse) at the right axilla near the armpit of each BALB / c nude mouse (18 - 20 g). Observe the growth of the transplanted tumor after injecting the tumor cells. Detect the diameter of the tumor mass with an electronic vernier caliper. When the transplanted tumor grows to a diameter of 0.5 cm or more, it is considered that the transplanted tumor nude mouse model is successfully established. Randomly divide 16 tumor-bearing mice into 2 groups, namely the Example 1 group and the control group. Raise the nude mice under SPF-level conditions, feed them with sterilized acidified water and sterile complete nutritional granular feed every day, and intragastrically administer a physiological saline suspension of 50 mg of the active ingredient / kg of the traditional Chinese medicine composition in Example 1 to the Example 1 group, and administer an equal amount of physiological saline to the control group. Administer the drug once a day for 15 consecutive days, record the tumor volume every 3 days, and draw a tumor growth curve. The results are as Figure 1 shown.

[0063] Figure 1 The results show that the example has a significant tumor inhibitory effect, indicating that the selected drug combination and structural design of the present invention can effectively treat tumors.

[0064] Experimental Example 2: BALB / c mice (18 - 20 g) were randomly divided into 4 groups, namely a control group, an Example 2 group, and Comparative Example 1 - 3 groups. They were orally administered normal saline, a normal saline suspension of the traditional Chinese medicine composition of Example 2 and Comparative Example 1 - 3, respectively, at a dosage of 50 mg active ingredient / kg, once every 2 days. The control group was given an equal amount of normal saline. At the same time, the body weight changes of the mice were monitored. The results are as Figure 2 shown.

[0065] Figure 2 The results showed that the body weights of the mice in the Example 2 and Comparative Example 1 - 3 groups increased slightly over time, similar to the control group, indicating that the traditional Chinese medicine composition of the present invention has good biocompatibility and can be used as a safe carrier for oral drug delivery.

[0066] Experimental Example 3: The traditional Chinese medicine compositions prepared in Example 3 and Comparative Example 1 - 3 were dispersed in PBS buffer solution containing 0.1% Tween 80 with a pH of 7.4 (ensuring that the content of the active ingredient in each group was the same), and oscillated at 100 rpm at 37°C for 24 h. 1 mL of the release medium was taken every 4 h to detect the content of the active ingredient quercetin, and new release medium was added. The cumulative release rate (%) at each time point was calculated. The results are as Figure 3 shown.

[0067] Figure 3 The results showed that the release rate of Example 3 was lower than that of Comparative Example 1 - 3. In Comparative Example 1, no cellulose - chitosan complex was added, and the release rate was faster. In Comparative Example 2, mesoporous SiO2 was used to replace the modified SiO2, and the release rate increased. In Comparative Example 3, chitosan was used to replace the modified chitosan, and the release rate increased. The above results showed that the cellulose - chitosan complex prepared in the present invention has a protective effect on the carrier, can prevent the premature release of the drug, and improve the sustained - release effect.

[0068] Experimental Example 4: SD rats were selected, and the feeding environment was maintained at a temperature of 25 ± 2°C, a relative humidity of 55%, and a normal light / dark cycle. 32 male SD rats (200 ± 20 g) were randomly divided into an Example 1 group and Comparative Example 1 - 3 groups, with 8 rats in each group. They were fasted but not water - deprived for 12 h before intragastric administration. Each group of rats was intragastrically administered a normal saline suspension of the samples of Example 1 and Comparative Example 1 - 3 at a dose of 50 mg / kg of quercetin concentration. After intragastric administration, blood was taken from the orbital venous plexus of the rats using a heparinized capillary and dropped into a heparinized EP tube. The blood sample was centrifuged at 3500 rpm for 10 min at 4°C, and the upper - layer plasma was aspirated to calculate the blood drug concentration of quercetin. After the last blood sampling was completed 24 h after intragastric administration, the area under the curve (AUC 0-t (mg·h·L -1 ) was calculated. The results are as Figure 4 shown.

[0069] Figure 4 The results showed that the AUC of the administration group of Example 1 after 24 h of administration was significantly higher than that of Comparative Examples 1-3, indicating that the absorption effect of the traditional Chinese medicine composition of Example 1 of the present invention in vivo was significantly better than that of Comparative Examples 1-3, and the oral bioavailability was better. It can prolong the action time in vivo, is beneficial to improving the bioavailability, has a longer action time, and enhances the therapeutic potential of the drug.

[0070] Those of ordinary skill in the art should understand that: the discussion of any above embodiment is only exemplary, and is not intended to imply that the scope of the present invention is limited to these examples; under the idea of the present invention, the technical features in the above embodiments or different embodiments can also be combined, the steps can be implemented in any order, and there are many other variations in different aspects of the present invention as described above, and they are not provided in detail for the sake of brevity.

Claims

1. A traditional Chinese medicine composition for treating tumors, characterized in that, The preparation raw materials include the following components in parts by weight: 3 - 6 parts of Chinese medicine extract, 5 - 10 parts of modified SiO₂, and 8 - 12 parts of cellulose-chitosan composite; The preparation method of the Chinese medicine extract includes the following steps: Take 20 - 30 parts of Scutellaria barbata, 20 - 30 parts of Hedyotis diffusa, 15 - 25 parts of Astragalus membranaceus, 10 - 20 parts of Codonopsis pilosula, 10 - 15 parts of Panax notoginseng, and 5 - 10 parts of Glycyrrhiza uralensis, dry and crush them, add them to an ethanol solution for soaking, reflux extraction, concentration, and freeze-drying to obtain the Chinese medicine extract; The preparation method of the modified SiO₂ includes the following steps: A1: Prepare mesoporous SiO₂ using CTAB, 2 M NaOH solution, and TEOS; A2: Modify the mesoporous SiO₂ with APTES to obtain amino-functionalized mesoporous SiO₂; A3: Modify the amino-functionalized mesoporous SiO₂ with succinic anhydride to obtain modified SiO₂; The preparation method of the cellulose-chitosan composite includes the following steps: B1: Modify 4 - 6 parts of CS with EDC·HCl and NHS to activate NAC to obtain modified chitosan; B2: Add 2 - 3 parts of cellulose to DMF and stir evenly, add 5 - 8 parts of thionyl chloride, react, cool, filter, dry, grind, add to distilled water, add 12 - 18 parts of 1,3-propanediamine, reflux, filter, wash the precipitate, and then dry it under vacuum; B3: Prepare an activation solution using NAC according to the method in step B1, add the product obtained after vacuum drying in step B2 to DMF, stir, add the activation solution, stir, dialyze, and freeze-dry to obtain modified cellulose; B4: Add the modified chitosan to ultrapure water and mix evenly, add 1 - 2 parts of 2,2'-dipyridyl disulfide to methanol and mix evenly, mix the two, react, centrifuge, wash, dry, disperse it with the modified cellulose in DMF, react, centrifuge, wash, and then dry to obtain the cellulose-chitosan composite.

2. The traditional Chinese medicine composition for treating tumors according to claim 1, wherein In step A1, the dosage ratio of CTAB, NaOH solution, and TEOS is 2 g:7 mL:10 mL; in step A2, the dosage ratio of mesoporous SiO₂ to APTES is 1 g:1 - 1.5 mL; in step A3, the mass ratio of amino-functionalized mesoporous SiO₂ to succinic anhydride is 1:4 - 6.

3. The traditional Chinese medicine composition for treating tumors according to claim 2, wherein, In step B1, the mass ratio of CS to NAC is 2:1; the mass ratio of NAC, EDC·HCl, and NHS is 2:5:

3.

4. The traditional Chinese medicine composition for treating tumors according to claim 3, wherein, In step B3, the mass ratio of NAC to cellulose is 1:

2.

5. The traditional Chinese medicine composition for treating tumors according to claim 4, wherein In step B4, the mass concentration of 2,2'-dipyridyl disulfide in methanol is 20 - 25 mg / mL; the mass concentration of modified chitosan in ultrapure water is 10 mg / mL; the mass concentration of modified cellulose in DMF is 10 mg / mL.

6. A method for preparing a traditional Chinese medicine composition for treating tumors according to any one of claims 1-5, characterized in that, It includes the following steps: S1: Disperse the modified SiO₂ and the Chinese medicine extract in ethanol, stir, centrifuge and wash, and dry to obtain drug-loaded modified SiO₂; S2: Add the drug-loaded modified SiO₂ obtained in step S1, EDC·HCl, and NHS to a PBS buffer solution with a pH of 5, stir, centrifuge, and collect the precipitate; S3: Disperse the precipitate obtained in step S2 in PBS buffer solution with a pH of 7.4, add a cellulose-chitosan complex, stir and react, centrifuge, wash the precipitate, and dry it to obtain a traditional Chinese medicine composition for treating tumors.

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