Synergistic and attenuated traditional Chinese medicine composition for osteosarcoma chemotherapy, preparation, preparation method and application

Abstract

The embodiment of the invention discloses an osteosarcoma chemotherapy effect-enhancing and toxicity-reducing traditional Chinese medicine composition, a traditional Chinese medicine preparation, a preparation method and application. The osteosarcoma chemotherapy effect-enhancing and toxicity-reducing traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 9 to 30 parts of radix astragali seu hedysari, 9 to 15 parts of radix scrophulariae, 5 to 10 parts of radix cyathulae, 6 to 12 parts of radix angelicae sinensis, 3 to 9 parts of pseudobulbus cremastrae seu pleiones and 9 to 30 parts of herba hedyotidis diffusae. The osteosarcoma chemotherapy efficacy-enhancing and toxicity-reducing traditional Chinese medicine composition provided by the embodiment of the invention consists of astragalus membranaceus, radix scrophulariae, radix cyathulae, angelica sinensis, edible tulip and oldenlandia diffusa, has a good curative effect on osteosarcoma, and can reduce the toxic and side effects of chemotherapeutic drugs on individuals with tumors.

Description

Technical Field

[0001] The present invention relates to the field of traditional Chinese medicine, and in particular to a traditional Chinese medicine composition for enhancing the efficacy and reducing the toxicity of osteosarcoma chemotherapy, a traditional Chinese medicine preparation, a preparation method and an application. Background Art

[0002] Osteosarcoma is a common bone malignancy in children and adolescents. It is prone to occur in the epiphyseal ends of long bones and is characterized by high malignancy, easy recurrence, and easy early metastasis. The current clinical treatment is mainly neoadjuvant chemotherapy combined with limb salvage surgery. Chemotherapy is the most commonly used treatment method, but patients face problems such as intolerance to chemotherapy drug doses and large toxic side effects. Therefore, the treatment of osteosarcoma is a major clinical problem.

[0003] In the field of traditional Chinese medicine, osteosarcoma belongs to the category of "stone carbuncle, stone carbuncle, bone carbuncle, stone rock" in traditional Chinese medicine. The pathogenesis is imbalance of yin and yang, insufficient kidney qi, dysfunction of viscera, accumulation of cold phlegm and dampness toxins, and stagnation of qi and blood in the bones. Phlegm and blood stasis are important pathological factors. Based on modern research, the TCM syndrome of osteosarcoma can be divided into qi deficiency and blood stasis type, kidney yang deficiency, cold stagnation of blood vessels type, kidney yin deficiency, weak tendons and atrophy of feet type, phlegm and blood stasis, malnutrition of tendons and bones type, yang deficiency and blood deficiency, cold stagnation of phlegm type. Traditional Chinese medicine has a rich theoretical basis and clinical experience in the treatment of osteosarcoma, and has the advantages of enhancing the efficacy and reducing the toxicity of chemotherapy, controlling the size of lesions, and preventing and treating distant metastasis.

[0004] Therefore, with modern pharmacological research methods, we conducted in-depth research and screening on the classic Chinese medicine prescriptions for treating cancer-related syndromes, and used data mining technology to extract and analyze the medication and prescription composition rules related to the treatment of osteosarcoma syndromes in the "Qing Palace Prescription Collection" by Academician Chen Keji, a master of traditional Chinese medicine. It is imperative to develop a drug combination to solve the problems of chemotherapy dose intolerance and large toxic side effects in osteosarcoma. Summary of the invention

[0005] Therefore, in order to overcome at least some of the defects and shortcomings in the prior art, the embodiments of the present invention provide a Chinese medicine composition, a Chinese medicine preparation, a preparation method and an application for enhancing the efficacy and reducing the toxicity of osteosarcoma chemotherapy.

[0006] On the one hand, an embodiment of the present invention provides a traditional Chinese medicine composition for enhancing the efficacy and reducing toxicity of osteosarcoma chemotherapy, wherein the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 9-30 parts of Astragalus, 9-15 parts of Scrophularia, 5-10 parts of Cyathula, 6-12 parts of Angelica, 3-9 parts of Psoralea corylifolia and 9-30 parts of Hedyotis diffusa.

[0007] In one embodiment of the present invention, the Chinese medicine composition comprises the following raw materials in parts by weight: 15 parts of Astragalus, 9 parts of Scrophularia, 9 parts of Cyathula, 9 parts of Angelica, 9 parts of Trillium and 12 parts of Hedyotis diffusa.

[0008] On the other hand, an embodiment of the present invention provides a Chinese medicine preparation of a Chinese medicine composition for enhancing the efficacy and reducing the toxicity of osteosarcoma chemotherapy, wherein the Chinese medicine preparation is prepared by adding a pharmaceutically acceptable excipient to the Chinese medicine composition.

[0009] In one embodiment of the present invention, the Chinese medicine preparation is an oral preparation, and the oral preparation is selected from the dosage form of decoction, extract, lyophilized powder, capsule, tablet, powder, granule, wine, pill and micropill.

[0010] In one embodiment of the present invention, the pharmaceutically acceptable excipient is selected from one or more of a diluent, a carrier, a filler, a binder, a wetting agent, a disintegrant, an absorption promoter, a surfactant, an adsorption carrier and a lubricant.

[0011] On the other hand, an embodiment of the present invention provides a method for preparing a traditional Chinese medicine composition for enhancing the synergy and reducing toxicity of osteosarcoma chemotherapy, comprising the following steps: weighing raw materials of Astragalus, Scrophularia, Cyathula, Angelica, Psoralea corylifolia and Hedyotis diffusa, adding ethanol aqueous solution, heating and refluxing, and filtering out the medicinal liquid to obtain a crude extract of the traditional Chinese medicine; concentrating the crude extract of the traditional Chinese medicine to obtain a concentrated solution of the traditional Chinese medicine; adding a pharmaceutically acceptable excipient to the concentrated solution of the traditional Chinese medicine to prepare an oral dosage form.

[0012] In one embodiment of the present invention, the step of adding an ethanol aqueous solution and heating under reflux and filtering out the medicinal liquid to obtain a crude Chinese medicine extract specifically comprises: adding an ethanol aqueous solution and heating under reflux, heating under reflux three times, each time for 2 hours, filtering out a first medicinal liquid, a second medicinal liquid and a third medicinal liquid, and mixing the first medicinal liquid, the second medicinal liquid and the third medicinal liquid to obtain the crude Chinese medicine extract.

[0013] In one embodiment of the present invention, the concentration of the ethanol aqueous solution is 70%.

[0014] In another aspect, an embodiment of the present invention provides the use of the above-mentioned Chinese medicine composition in the preparation of a chemotherapy synergistic and toxicity-reducing drug for osteosarcoma.

[0015] In summary, the traditional Chinese medicine composition for enhancing the efficacy and reducing toxicity of osteosarcoma chemotherapy proposed in the embodiment of the present invention is composed of Astragalus, Scrophularia, Cyathula, Angelica, Trillium and Oldenlandia diffusa, so that the traditional Chinese medicine composition proposed in the embodiment of the present invention has a good therapeutic effect on osteosarcoma and can reduce the toxic and side effects of chemotherapy drugs on tumor-bearing individuals. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other accompanying drawings can be obtained based on these accompanying drawings without paying creative work.

[0017] Figure 1 The present invention provides a flow chart of a method for preparing a traditional Chinese medicine composition for enhancing the chemotherapy efficacy and reducing the toxicity of osteosarcoma provided in an embodiment of the present invention.

[0018] Figure 2 This is a comparison chart of liver cells from mice in the maximum dose experiment.

[0019] Figure 3A and Figure 3B This is a comparison chart of kidney cells from mice in the maximum dosage experiment.

[0020] Figure 4 It is a line graph showing the effect of the Chinese medicine composition on the survival rate of osteosarcoma 143B cells.

[0021] Figure 5 It is a line graph of the changes in tumor volume in the combination drug experiment.

[0022] Figure 6 This is a comparison chart of tumor weight in the combined drug experiment.

[0023] Figure 7 This is a comparison chart of mouse body weight in the combined drug experiment. DETAILED DESCRIPTION

[0024] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. 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.

[0025] The embodiment of the present invention provides a traditional Chinese medicine composition for enhancing the efficacy and reducing toxicity of osteosarcoma chemotherapy. The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 9-30 parts of astragalus, 9-15 parts of Scrophularia, 5-10 parts of Cyathula, 6-12 parts of Angelica, 3-9 parts of Psoralea corylifolia and 9-30 parts of Hedyotis diffusa.

[0026] In a specific implementation of this embodiment, the Chinese medicine composition includes the following raw materials in parts by weight: 15 parts of Astragalus, 9 parts of Scrophularia, 9 parts of Cyathula, 9 parts of Angelica, 9 parts of Trillium and 12 parts of Hedyotis diffusa.

[0027] The embodiment of the present invention provides a Chinese medicine preparation of a Chinese medicine composition for enhancing the efficacy and reducing toxicity of osteosarcoma chemotherapy, wherein the Chinese medicine preparation is prepared by adding a pharmaceutically acceptable excipient to the Chinese medicine composition. The Chinese medicine preparation is an oral preparation, and the oral preparation is selected from the dosage form of a decoction, an extract, a lyophilized powder, a capsule, a tablet, a powder, a granule, an alcoholic preparation, a pill, and a micropill. Among them, the pharmaceutically acceptable excipient is selected from one or more of a diluent, a carrier, a filler, a binder, a wetting agent, a disintegrant, an absorption promoter, a surfactant, an adsorption carrier, and a lubricant.

[0028] See also Figure 1 The present invention also provides a method for preparing the above-mentioned Chinese medicine composition, comprising the following steps:

[0029] S10, weighing astragalus, Scrophularia, Cyathula, Angelica, Psoralea corylifolia and Hedyotis diffusa raw materials, adding ethanol aqueous solution, heating under reflux, and filtering out the medicinal solution to obtain a crude extract of traditional Chinese medicine;

[0030] S20, concentrating the crude extract of traditional Chinese medicine to obtain a concentrated solution of traditional Chinese medicine;

[0031] S30, adding a pharmaceutically acceptable excipient to the traditional Chinese medicine concentrate to prepare an oral dosage form.

[0032] In step S10, after adding the ethanol-water solution and heating under reflux, filtering out the medicinal liquid to obtain a crude Chinese medicine extract specifically includes: adding the ethanol-water solution and heating under reflux, heating under reflux three times, heating under reflux for 2 hours each time, filtering out a first medicinal liquid, a second medicinal liquid and a third medicinal liquid, and mixing the first medicinal liquid, the second medicinal liquid and the third medicinal liquid to obtain the crude Chinese medicine extract.

[0033] Preferably, the concentration of the ethanol aqueous solution is 70%, and the amount of the ethanol aqueous solution is 10 times the amount of the medicinal material.

[0034] In order to facilitate a better understanding of the embodiments of the present invention, the following experimental description is given by taking the preparation of a Chinese medicine composition for enhancing the synergy and reducing toxicity of osteosarcoma chemotherapy into a powdered Chinese medicine as an example.

[0035] [Example 1]

[0036] 9 parts of Astragalus, 9 parts of Scrophularia, 5 parts of Cyathula, 6 parts of Angelica, 3 parts of Psoralea corylifolia and 9 parts of Hedyotis diffusa.

[0037] [Example 2]

[0038] 30 parts of Astragalus, 15 parts of Scrophularia, 10 parts of Cyathula, 12 parts of Angelica, 9 parts of Psoralea corylifolia and 30 parts of Oldenlandia diffusa.

[0039] [Example 3]

[0040] 15 parts of Astragalus, 9 parts of Scrophularia, 9 parts of Cyathula, 9 parts of Angelica, 9 parts of Psoralea corylifolia and 12 parts of Hedyotis diffusa.

[0041] Preparation process of powdered Chinese medicine composition for enhancing efficacy and reducing toxicity of osteosarcoma chemotherapy:

[0042] (1) Weigh the Chinese medicinal materials required for each dose of the Chinese medicinal compound composition, add 10 times the weight of the raw materials to a 70% ethanol aqueous solution, soak for 30 minutes, heat and reflux for 2 hours, and filter out the first medicinal liquid; then add 8 times the weight of the 70% ethanol aqueous solution, heat and reflux for 2 hours, and filter out the second medicinal liquid; then add 8 times the weight of 70% ethanol, heat and reflux for 2 hours, and filter out the third medicinal liquid, and mix the first medicinal liquid, the second medicinal liquid and the third medicinal liquid to obtain a crude extract.

[0043] (2) The crude extract obtained in step (1) is concentrated by rotary evaporation until it becomes thick to obtain a concentrated solution, which can be passed through a 12-gauge gavage needle.

[0044] (3) freeze-drying the concentrated solution obtained in step (2) under vacuum for more than 36 hours to obtain a powdered Chinese medicine compound extract, i.e., a lyophilized powder of the Chinese medicine compound composition, wherein each gram of lyophilized powder is equivalent to 2.291 g of the crude drug.

[0045] Drug toxicology experiments:

[0046] The raw materials of the embodiments of the present invention include Astragalus, Scrophularia, Cyathula, Angelica, Psoralea corylifolia, and Hedyotis diffusa. To facilitate clinical application, their safety was evaluated, and thus a toxicology experiment was designed.

[0047] Drug acute toxicity test:

[0048] Experimental animals: 20 ICR mice (SYXK (Shanghai) 2022-0012), SPF grade, half male and half female, weighing 20±2 g, were raised in the Experimental Animal Center of Fujian University of Traditional Chinese Medicine (SYXK (Min) 2019-0007). Experimental intervention was performed after 1 week of adaptive feeding.

[0049] Administration: Oral administration, the administration volume was 0.4mL / 10gb.w, the administration group was given the above extract, and the control group was given normal saline, and the observation was continued for 14 days.

[0050] Observation contents: general activity status, morphological signs, diet, stool, body weight, and morphology of liver, heart, spleen, lungs, and kidneys; the time of onset, manifestation, aggravation, relief, and death of toxic reactions were recorded.

[0051] Experimental results: After administration, the mice had normal fur color, normal morphology, normal activities, started eating, had no abnormal secretions or exudation from the eyes and ears, had dry and formed stools, and no deaths were observed. Gross anatomical observation of the liver, heart, spleen, lungs, kidneys and other major organs showed no abnormalities in color, shape, size, etc.

[0052] Maximum dosage experiment:

[0053] Due to the limited stomach capacity of mice, the lethal dose for mice was not measured, so the maximum dosage was further tested.

[0054] Experimental animals: 40 ICR mice. Other procedures were the same as those in the acute toxicity test.

[0055] Administration: Same as acute toxicity test.

[0056] Observation contents: serum was collected for blood biochemical detection of liver and kidney function related indicators, liver and kidney tissues were collected for pathological sections, and the rest was the same as the acute toxicity test.

[0057] Experimental results: After administration, the mice had normal fur color, normal morphology, normal activities, started to eat, had no abnormal secretions and exudation from the eyes and ears, had dry and formed stools, and no deaths. Gross autopsy and naked eye observation of the liver, heart, spleen, lungs, kidneys and other major organs showed no abnormalities in color, shape, size, etc. See Table 1. Compared with the control group, there was no significant difference in the weight of mice in the administration group. See Table 2. There was no significant difference in the weight of the liver, heart, spleen, lungs, and kidneys of mice. See Table 2. There was no significant difference in the liver and kidney function indicators of mice. See Figure 2 , Figure 3A and Figure 3B , no obvious pathological changes were observed in the liver and kidney of mice. It can be concluded from the above experimental results that the Chinese medicine composition provided in this embodiment has no pathological damage to the liver and kidney of mice, and has no obvious toxic side effects on mice.

[0058] Table 1 Changes in body weight of mice at the maximum dose

[0059]

[0060] Table 2 Maximum dose mouse liver, heart, spleen, lung and kidney weight (unit: g, ±s)

[0061]

[0062] Table 3 Detection of serum liver and kidney function in mice in the maximum dosage experiment

[0063]

[0064]

[0065] Effect of a traditional Chinese medicine composition for enhancing the efficacy and reducing toxicity of osteosarcoma chemotherapy on IC of osteosarcoma cells 50 (half maximal inhibitory concentration) determination:

[0066] CCK-8 method to detect cell viability: 143B cells with good growth status were collected according to the subculture method and 5×10 4 The cells were inoculated at a density of 100 μL per well in a 96-well plate at 37°C and 5% CO2 incubator. The cells were observed under a microscope. When the cell growth confluence reached 40-50%, the culture medium was replaced with complete culture medium and culture medium containing 700 μg, 800 μg, 900 μg, 1 mg, and 1.1 mg of the Chinese medicine compound for 48 hours. Six replicates were set for each concentration, and a normal culture medium and no cells were set as a blank group. After the intervention of each group, 10 μL of CCK-8 reagent and 90 μL of blank culture medium were added to each well in turn under light-proof conditions to avoid bubbles. After incubation in a 37°C incubator for 2 hours, the OD value was measured at a wavelength of 450 nm using an enzyme reader, and the average value of each replicate was taken. The experiment was repeated 3 times.

[0067] The experimental results are as follows Figure 4 As shown, the osteosarcoma chemotherapy synergistic and attenuated Chinese herbal composition provided in this embodiment can inhibit the survival rate of osteosarcoma cells, and there is a dose-effect relationship between the osteosarcoma chemotherapy synergistic and attenuated Chinese herbal composition and the survival rate of osteosarcoma cells, and the 24h, 48h, and 72h half-inhibition rates of osteosarcoma 143B cells are 1.339, 1.069, and 0.799 mg / mL.

[0068] Evaluation of the efficacy of osteosarcoma chemotherapy-enhancing and -reducing Chinese medicine composition combined with chemotherapy drugs on osteosarcoma:

[0069] Construction of osteosarcoma cell transplant tumor model in Balb / c nude mice: 40 Balb / c nude mice were used to obtain 143B cells with good growth status. After the cells grew to 90%, they were digested with trypsin and collected, and resuspended in matrix gel. The cell concentration was adjusted to 1×10 7 / mL, and 0.2mL / mouse was inoculated into the subcutaneous part of the right forelimb of Balb / c nude mice to establish an osteosarcoma nude mouse transplant tumor model. After the tumor was formed, the mice were divided into control group, Chinese medicine compound group, and chemotherapy drug combination Chinese medicine compound combined with chemotherapy drug group for intervention for 14 days. During the period, the weight and tumor volume of the mice were measured, and the tumor was weighed after the experiment.

[0070] Experimental results: see Figure 5 and Figure 6 It can be clearly seen that the Chinese medicine composition can inhibit the growth of osteosarcoma nude mouse transplanted tumors; compared with the treatment with chemotherapy drugs alone, the Chinese medicine composition combined with chemotherapy drugs (such as cisplatin) has a more significant inhibitory effect on the growth of mouse transplanted tumors. Figure 7 It can be seen that compared with the use of chemotherapy drugs alone, the combination of the Chinese medicine composition and chemotherapy drugs can increase the body weight of tumor-bearing mice.

[0071] In summary, the osteosarcoma chemotherapy synergistic and toxicity-reducing traditional Chinese medicine composition provided in the embodiment of the present invention is composed of Astragalus, Scrophularia, Cyathula, Angelica, Trillium and Oldenlandia diffusa, so that the traditional Chinese medicine composition proposed in the embodiment of the present invention has a good therapeutic effect on osteosarcoma and can reduce the toxic and side effects of chemotherapy drugs on tumor-bearing individuals.

[0072] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A Chinese medicine composition for enhancing the efficacy and reducing the toxicity of osteosarcoma chemotherapy, characterized in that: The Chinese medicine composition comprises the following raw materials in parts by weight: 9-30 parts of astragalus, 9-15 parts of figwort, 5-10 parts of cyathula, 6-12 parts of angelica, 3-9 parts of trillium and 9-30 parts of oldenlandia diffusa.

2. The osteosarcoma chemotherapy efficacy-enhancing and toxicity-reducing traditional Chinese medicine composition according to claim 1, characterized in that: The Chinese medicine composition comprises the following raw materials in parts by weight: 15 parts of astragalus, 9 parts of figwort, 9 parts of cyathula, 9 parts of angelica, 9 parts of trillium and 12 parts of oldenlandia diffusa.

3. A Chinese medicine preparation of the osteosarcoma chemotherapy synergistic and toxicity-reducing Chinese medicine composition according to claim 1 or 2, characterized in that: The traditional Chinese medicine preparation is prepared by adding a pharmaceutically acceptable excipient to the traditional Chinese medicine composition.

4. The Chinese medicine preparation according to claim 3, characterized in that: The Chinese medicine preparation is an oral preparation, and the oral preparation is selected from the dosage form of decoction, extract, lyophilized powder, capsule, tablet, powder, granule, wine, pill and micropill.

5. The Chinese medicine preparation according to claim 3, characterized in that: The pharmaceutically acceptable excipient is selected from one or more of a diluent, a carrier, a filler, a binder, a wetting agent, a disintegrant, an absorption promoter, a surfactant, an adsorption carrier and a lubricant.

6. A method for preparing the Chinese medicine composition according to claim 1 or 2, characterized in that: The following steps are involved: Weigh astragalus, Scrophularia, Cyathula, Angelica, Psoralea corylifolia and Hedyotis diffusa raw materials, add ethanol aqueous solution, heat under reflux, filter out the medicinal solution, and obtain a crude extract of traditional Chinese medicine; Concentrating the crude extract of traditional Chinese medicine to obtain a concentrated solution of traditional Chinese medicine; A pharmaceutically acceptable excipient is added to the traditional Chinese medicine concentrated solution to prepare an oral dosage form.

7. The preparation method according to claim 6, characterized in that: The ethanol aqueous solution is added, heated under reflux, and the medicinal solution is filtered out to obtain a crude extract of traditional Chinese medicine, which specifically comprises: Add ethanol aqueous solution and heat to reflux, heat to reflux three times, each time for 2 hours, filter out to obtain the first medicinal liquid, the second medicinal liquid and the third medicinal liquid, mix the first medicinal liquid, the second medicinal liquid and the third medicinal liquid to obtain the crude Chinese medicine extract.

8. The preparation method according to claim 7, characterized in that: The concentration of the ethanol aqueous solution is 70%.

9. The preparation method according to claim 7, characterized in that: The amount of the ethanol aqueous solution is 10 times the amount of the medicinal material.

10. Use of the Chinese medicine composition according to claim 1 or 2 in the preparation of a drug for enhancing the efficacy and reducing the toxicity of chemotherapy for osteosarcoma.

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