Traditional Chinese medicine composition for preventing and treating myelosuppression after chemotherapy

Abstract

The invention belongs to the field of traditional Chinese medicines, and particularly discloses a traditional Chinese medicine composition for preventing and treating myelosuppression after chemotherapy. The composition is prepared from radix scrophulariae, bighead atractylodes rhizome, astragalus membranaceus, sargentgloryvine stem, radix rehmanniae recen, angelica sinensis, hairyvein agrimony, caulis spatholobi, dandelion and eclipta. Pharmacodynamic experiments show that the composition can improve platelet count of mice with platelet reduction caused by cyclophosphamide or carboplatin, and has the effect of promoting generation of megakaryocyte of mouse bone marrow. The composition disclosed by the invention is definite in curative effect and small in side effect, has a remarkable effect on myelosuppression caused by chemotherapy, especially on prevention and treatment of thrombocytopenia after chemotherapy, and enriches clinical medication selection of tumor patients.

Description

Technical Field

[0001] The invention belongs to the field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition for preventing and treating post-chemotherapy bone marrow suppression and a preparation method thereof. Background Art

[0002] With the development of society and economy, human life expectancy has been extended overall, and malignant tumors are one of the serious diseases that threaten human life and health. Surgery, radiotherapy, and chemotherapy are the main methods for treating malignant tumors. Chemotherapy is a way of using chemical drugs to prevent the proliferation, infiltration, and metastasis of cancer cells until the cancer cells are finally killed. It is the most common means of anti-tumor treatment. Most patients can achieve relatively ideal treatment effects through chemotherapy, prolong survival time, and delay the development of the disease. However, due to the low selectivity of chemotherapy drugs, they mainly target actively growing cells. In addition to malignant tumor cells, cells in organs such as bone marrow hematopoietic stem cells, digestive tract mucosa, skin and appendages, endometrium, and ovaries are also updated rapidly, often resulting in a series of toxic and side effects such as bone marrow suppression, gastrointestinal reactions, and liver and kidney damage. Among them, bone marrow suppression seriously affects the treatment process of tumor patients, thereby affecting the anti-tumor effect.

[0003] Chemotherapy-induced bone marrow suppression manifests as a decrease in white blood cells, platelets, or red blood cells; among them, chemotherapy-induced thrombocytopenia (CIT) is the most difficult to treat. Thrombocytopenia caused by anti-tumor drugs will increase the patient's bleeding risk and prolong hospitalization. Clinicians often choose to reduce the chemotherapy dose, extend the chemotherapy interval, or even terminate chemotherapy, which affects the efficacy and prognosis. Therefore, the prevention and treatment of thrombocytopenia has become a difficult point in tumor treatment.

[0004] At present, Western medicine treatment of CIT mostly uses platelet transfusion, subcutaneous injection of recombinant human thrombopoietin (rhTPO), recombinant human interleukin-11 (rhIL-11) and other drugs that promote platelet regeneration. These drugs have certain effects in promoting platelet production, reducing bleeding risks, and ensuring that chemotherapy is carried out as scheduled and in sufficient quantities. However, they are consumed rapidly, have a short maintenance period, and have many adverse reactions.

[0005] As a special feature of tumor treatment in my country, traditional Chinese medicine has certain advantages and clinical efficacy in shrinking lesions, reducing the toxic side effects of chemotherapy, improving the quality of life, and prolonging the overall survival period. In recent years, traditional Chinese medicine has been widely used in the prevention and treatment of adverse reactions to tumor chemotherapy, and has a significant therapeutic effect on CIT.

[0006] At present, each medical institution of TCM for the prevention and treatment of CIT has its own prescriptions, which have certain efficacy within the statistical range, but there are also some shortcomings. For example, the prescriptions formed under the guidance of syndrome differentiation are too large, there is no clear legislative rule, and the main purpose of strengthening the spleen, warming the kidney, nourishing yin, and replenishing qi is unclear; the complexity of tumor treatment, the etiology and pathogenesis of bone marrow suppression caused by it are different, and it is difficult to determine a clear treatment plan. In view of the above problems, it is urgent to provide a Chinese medicine combination with clear efficacy to enrich the clinical drug selection. Summary of the invention

[0007] In order to enrich the medication options for myelosuppression after chemotherapy, the present invention provides a Chinese medicine composition with clear efficacy, which is used for tumor patients to resist the side effects of myelosuppression during chemotherapy, especially has a significant efficacy for thrombocytopenia after chemotherapy.

[0008] The first aspect of the present invention provides a traditional Chinese medicine composition for preventing and treating bone marrow suppression after chemotherapy, which is made from the following traditional Chinese medicine components: Scrophularia ningpoensis, Atractylodes macrocephala, Astragalus membranaceus, Sargentodoxae, Rehmannia root, Angelica sinensis, Agrimoniae, Millettia spatholobi, Taraxacum mongolicum and Eclipta prostrata.

[0009] Preferably, the Chinese medicine composition is made of the following Chinese medicine components in parts by weight:

[0010]

[0011] Preferably, the Chinese medicine composition is composed of the following Chinese medicine components in parts by weight:

[0012]

[0013] In a preferred embodiment, the Chinese medicine composition is made of the following Chinese medicine components in parts by weight:

[0014]

[0015] Traditional Chinese medicine believes that chemotherapy drugs are offensive, consume Qi and damage Yin, and belong to the category of "fire poison" and "fire and heat poison" of the six exogenous evils. Multiple cycles of chemotherapy will damage the body's own blood system and immune system, causing the body to be in a pathological state of "blood deficiency". Blood is one of the most basic nutrients for the human body. If the blood is deficient, the nourishing and metaplastic functions of the blood will be low, and the human organs and limbs will not be nourished, causing the body to become "more deficient". After chemotherapy, when thrombocytopenia occurs due to bone marrow suppression, patients are often accompanied by dizziness, fatigue, tiredness, poor appetite, soreness of the limbs, etc. In severe cases, there may be subcutaneous mucosal bleeding, and even critical and severe bleeding of important organs, which belongs to the category of "deficiency".

[0016] In the composition of the present invention, Scrophularia ningpoensis nourishes yin and cools blood, Astragalus invigorates qi and nourishes blood as the main drugs, Atractylodes macrocephala strengthens the spleen and invigorates qi, Radix Rehmanniae and Caulis Sargentodoxae clear away heat and cool blood, Angelica sinensis and Caulis Spatholobi nourish blood and activate blood circulation, which are all assistant drugs, helping the main drugs to play the functions of nourishing blood and cooling blood; Herba Agrimoniae astringes and stops bleeding, Herba Taraxaci clears heat and disperses stagnation, and Herba Ecliptae cools blood and stops bleeding as the adjuvant drugs. All the drugs work together to achieve the effects of nourishing yin and invigorating qi, nourishing blood and cooling blood.

[0017] The second aspect of the present invention provides a method for preparing the Chinese medicine composition, wherein the Chinese medicine raw materials of the above formula are extracted or processed in other ways to prepare a pharmaceutically active substance, and the substance is used as a raw material, and pharmaceutically acceptable excipients are added when necessary to prepare various pharmaceutically acceptable dosage forms according to conventional techniques of pharmaceutics. The active substance is obtained by extracting the Chinese medicine raw materials separately, extracting the Chinese medicine raw materials together, or other methods, such as: crushing, soaking, extraction, percolation, distillation, decoction, concentration, alcohol precipitation, water precipitation, centrifugation, filtration, chromatography, drying, sieving, etc. These active substances can be substances in the form of extracts, which can be dry extracts or fluid extracts, and different concentrations can be prepared according to different preparation requirements.

[0018] The preferred processing method is as follows: the medicinal material is decocted with water for 1-3 times, each time for 1-3 hours, and 6-12 times the amount of water is added each time. The decoction is filtered, combined, and concentrated under reduced pressure to an extract with a relative density of 1.0-1.4 (50-60° C.), and placed at room temperature. If necessary, ethanol is added to make the alcohol content reach 60%-80%, and the extract is fully stirred, allowed to stand, filtered, and the filtrate is concentrated under reduced pressure to recover the ethanol. The concentrate is added with pharmaceutically acceptable excipients to prepare an oral liquid preparation or dried into a dry powder and then added with pharmaceutically acceptable excipients to prepare an oral solid preparation.

[0019] The preparation method of the Chinese medicine composition comprises the following steps:

[0020] (1) Remove impurities from Scrophularia and Angelica, wash, soak, cut into thin slices, and set aside; remove impurities from Atractylodes, Sargentodoxae, and Radix Rehmanniae, wash, soak, cut into thick slices, dry, and set aside; remove impurities from Agrimonia, Taraxacum, and Eclipta prostrata, wash, cut into sections, dry, and set aside;

[0021] (2) taking the ten medicinal materials processed in step (1) in the prescribed amount, adding 6-12 times of water and boiling and extracting for 2-3 times, each time for 1-3 hours; filtering, collecting and combining the decoction, and concentrating under reduced pressure to a relative density of 1.05-1.4 (50-60° C.), to obtain an extract;

[0022] (3) adding pharmaceutically acceptable excipients to the extract obtained in step (2) and preparing the extract into a clinically acceptable dosage form through conventional processes.

[0023] Preferably, the pharmaceutically acceptable excipient is one or more of a filler, a diluent, a flavoring agent, a lubricant, a preservative, a binder, a disintegrant, and a preservative.

[0024] Further preferably, the pharmaceutically acceptable excipients include but are not limited to lactose, starch, dextrin, powdered sugar, magnesium stearate, maltose, lemonade, tartaric acid, sodium hydroxide, aspartame, stevioside, cyclamate, protein sugar, acesulfame potassium, aspartame, sucralose and the like.

[0025] Preferably, the clinically acceptable dosage form is granules, tablets, pills, capsules, oral solutions or mixtures.

[0026] In one specific embodiment, the Chinese medicine composition is a granule, which is prepared by the following method:

[0027] (1) Remove impurities from Scrophularia and Angelica, wash, soak, cut into thin slices, and set aside; remove impurities from Atractylodes, Sargentodoxae, and Radix Rehmanniae, wash, soak, cut into thick slices, dry, and set aside; remove impurities from Agrimonia, Taraxacum, and Eclipta prostrata, wash, cut into sections, dry, and set aside;

[0028] (2) taking the ten medicinal materials processed in step (1) in the prescribed amount, adding 6-12 times of water and boiling and extracting for 2-3 times, each time for 1-3 hours; filtering, collecting and combining the decoctions, and concentrating under reduced pressure to an extract having a relative density of 1.1-1.4 at 50° C.-60° C.;

[0029] (3) The extract obtained in step (2) is vacuum dried to obtain a dry paste, and pharmaceutically acceptable excipients are added, dry granulated or wet granulated, granulated, and dried to obtain granules.

[0030] In another specific embodiment, the Chinese medicine composition is an oral liquid preparation, which is prepared by the following method:

[0031] (1) Remove impurities from Scrophularia and Angelica, wash, soak, cut into thin slices, and set aside; remove impurities from Atractylodes, Sargentodoxae, and Radix Rehmanniae, wash, soak, cut into thick slices, dry, and set aside; remove impurities from Agrimonia, Taraxacum, and Eclipta prostrata, wash, cut into sections, dry, and set aside;

[0032] (2) taking the ten medicinal materials processed in step (1) in the prescribed amount, adding 6-12 times of water and boiling and extracting for 2-3 times, each time for 1-3 hours; filtering, collecting and combining the decoctions, and concentrating under reduced pressure to an extract having a relative density of 1.05-1.15 at 50-60° C.;

[0033] (3) adding ethanol to the extract obtained in step (2) until the alcohol content is 60%-80%, cooling and settling for 24-48 hours, filtering and obtaining the alcohol precipitate for later use;

[0034] (4) recovering ethanol by vacuum concentration and concentrating to obtain an extract or a liquid for preparation; adding a flavoring agent and / or a preservative, adjusting the pH, adding water to the total amount of preparation, stirring, filtering, filling, and sterilizing to obtain an oral liquid preparation.

[0035] The Chinese medicine composition of the present invention is used in the preparation of drugs for preventing and treating bone marrow suppression after chemotherapy, wherein the bone marrow suppression is manifested as a decrease in white blood cells, red blood cells or platelets after chemotherapy.

[0036] The Chinese medicine composition of the invention has clear curative effect and small side effect, and can prevent and treat bone marrow suppression after chemotherapy, and has a significant effect in preventing and treating thrombocytopenia after chemotherapy.

[0037] In order to verify the efficacy of the Chinese medicine composition of the present invention, the inventors carried out pharmacodynamic tests. It should be noted that the drugs selected for the pharmacodynamic tests of the present invention are drugs obtained by the representative formulas and preparation methods of the present invention. Due to space limitations, the tests and results involved in the drugs obtained by other formulas and preparation methods included in the present invention are not listed here.

[0038] At present, patients with thrombocytopenia caused by chemotherapy are mainly treated with rhTPO or rhIL-11. Clinical data show that in the process of chemotherapy for tumor patients, the application of rhTPO or rhIL-11 can effectively reduce the degree of thrombocytopenia and accelerate platelet recovery, thereby reducing the patient's demand for platelet transfusion. Although the efficacy is certain, it requires high costs and has certain side effects. Long-term injection will cause the body to produce antibodies, affecting the treatment effect. The Chinese medicine composition of the present invention has the effects of invigorating qi and nourishing blood, promoting blood circulation and cooling blood, nourishing yin and tonifying the kidney, can prevent and treat the level of platelet reduction after chemotherapy, and improve the quality of life of patients.

[0039] Experimental Example 1 Effect of the composition of the present invention on thrombocytopenia in mice induced by cyclophosphamide

[0040] 1. Experimental Materials

[0041] Reagents and instruments: The composition of the present invention is the granules prepared in Example 1, recombinant human thrombopoietin injection (rhTPO), specifications: 1 ml: 15,000 units, produced by Shenyang Sansheng Pharmaceutical Co., Ltd.; cyclophosphamide (cytoxan, CTX): 0.2 g / vial, produced by Jiangsu Hengrui Medicine Co., Ltd.; mouse thrombopoietin ELISA kit, produced by Wuhan Boster Bioengineering Co., Ltd.; Sysmex-K21 blood cell analyzer, Jinan Sysmex Medical Electronics Co., Ltd.

[0042] Experimental animals: 60 Kunming mice, male, 6-8 weeks old, body weight 18-22 g, experimental animal license number SYXK (Lu) 20180008, provided by Lunan Pharmaceutical Group Co., Ltd., adaptive feeding for one week before the experiment, free access to food and water during the experiment, 12 h circadian rhythm.

[0043] 2. Experimental Methods

[0044] 2.1 Experimental Animal Grouping and Dosing

[0045] Kunming mice were adapted to feeding in the laboratory for 2 days, blood routine was measured, and then they were randomly divided into 6 groups according to platelet count and body weight balance, with 10 mice in each group. They were blank control group, model control group, positive control group, low-dose group, medium-dose group, and high-dose group. Administration method: the model control group, low-dose group, medium-dose group, and high-dose group of the present composition were respectively administered with equal volumes of normal saline, 8.4g raw medicinal materials / kg, 16.7g raw medicinal materials / kg, and 32.3g raw medicinal materials / kg of the present composition 5 days before modeling; once a day, after CTX modeling on the 6th day, continuous daily gavage for 6 days; the positive control group began to subcutaneously inject recombinant thrombopoietin rhTPO (2000U) daily 1h before modeling for 6 consecutive days, and the blank control group was only given an equal volume of normal saline gavage.

[0046] 2.2 Modeling method

[0047] Except for the blank control group, mice in each group were intraperitoneally injected with cyclophosphamide CTX 100 mg / kg 1 hour after administration on the 6th day, once a day for 3 consecutive days to complete the model establishment. The platelet count (PLT) decreased to 20%-30% before administration as the standard for successful model establishment.

[0048] 2.3 Index observation and measurement

[0049] 2.3.1 Peripheral blood cell count

[0050] 20 μl of blood was collected from the tail vein of each group 2 hours after administration before the experiment (d1), before modeling (d6), 3 days (d9) and 6 days (d12) after modeling, and the platelet count and mean platelet volume (MPV) were determined using a cell analyzer after dilution.

[0051] 2.3.2 Determination of bone marrow megakaryocyte count

[0052] On the 12th day of the experiment, mice were killed by cervical dislocation 2 hours after administration. The femurs on both sides of the mice were freed, and 1 ml of PBS buffer was drawn with a 1 ml syringe to flush out all the bone marrow cells in the femurs. The cell suspension was passed through a syringe with a No. 4 needle to fully disperse the cells in the suspension. After centrifugation and aspiration of the bone marrow supernatant, the bone marrow cell fluid was smeared and stained, and the number of all bone marrow megakaryocytes was counted under a microscope using the white blood cell counting method.

[0053] 2.3.3 TPO detection in serum and bone marrow supernatant: On the 12th day of the experiment, 1 ml of blood was collected from the femoral vein (0.1 ml of heparin was added to the test tube for anticoagulation), centrifuged, and serum was separated; bone marrow supernatant was prepared according to the method in 2.3.2. The TPO content in serum and bone marrow supernatant was detected by ELISA according to the instructions. The OD was measured at 450 nm using a Spectra Max M2 multifunctional microplate reader, and the OD curve was drawn to obtain the TPO concentration of the corresponding sample.

[0054] 2.3.4 Data processing methods

[0055] The experimental data were statistically analyzed using SPSS22 software, and the data were expressed as mean ± standard deviation. The results were analyzed by one-way analysis of variance, and the t-test was used to compare the means of the two groups. P < 0.01 and P < 0.05 indicated that the difference between the two groups was statistically significant.

[0056] 3 Experimental results

[0057] 3.1 Comparison of PLT counts in each group of mice

[0058] Platelet count (PLT) is an indicator that reflects platelet production and death. The effects of the composition of the present invention on the PLT counts of mice in each group are shown in Table 1. From the results in Table 1, it can be seen that the PLT counts of mice in the model group on the 9th day were significantly lower than those in the blank control group (P < 0.05), indicating that the mouse model was successfully established. The platelet counts of mice in each medication group on the 12th day were significantly higher than those of mice in the model control group (P < 0.05, P < 0.01). This indicates that both the positive control drug and the composition of the present invention can increase the peripheral blood PLT counts of mice with platelet reduction caused by cyclophosphamide.

[0059] Table 1 Changes in peripheral blood PLT counts of mice in each group ( n=10)

[0060]

[0061] Note: Compared with the blank control group, "*" indicates P < 0.05; compared with the model control group, ▲ "P<0.05," ▲▲ ” indicates P<0.01.

[0062] 3.2 Comparison of mean platelet volume (MPV) in each group of mice

[0063] Mean platelet volume (MPV) represents the volume of platelets. It is a parameter that reflects the proliferation, metabolism and platelet production of megakaryocytes in the bone marrow, and also shows the age of circulating platelets. An increase in MPV indicates that the maturity of megakaryocytes is high, and vice versa. The newly generated platelets are larger in size, have more dense bodies, higher enzyme activity, and stronger functions, which is manifested by an increase in MPV. The effects of the composition of the present invention on the peripheral blood MPV of each group of mice are shown in Table 2. From the results in Table 2, it can be seen that on the 9th day, the peripheral blood MPV of the mice in the model group was significantly higher than that of the blank control group (P < 0.05). On the 12th day, the peripheral blood MPV of the mice in the medium and high dose groups was significantly higher than that of the model control group (P < 0.05, P < 0.01). It shows that the composition of the present invention can increase the peripheral blood MPV count of mice with platelet reduction caused by cyclophosphamide.

[0064] Table 2 Changes in peripheral blood MPV counts of mice in each group ( n=10)

[0065]

[0066] Note: Compared with the blank control group, "*" indicates P < 0.05; compared with the model control group, ▲ "P<0.05," ▲▲ ” indicates P<0.01.

[0067] 3.3 Determination of TPO concentration in serum and bone marrow supernatant of mice in each group

[0068] TPO acts directly on bone marrow hematopoietic stem cells and regulates all stages of platelet production. TPO is an endogenous cytokine that stimulates the growth and differentiation of megakaryocytes. It has a stimulating effect on all stages of megakaryocyte production, including the proliferation of precursor cells and the maturation of polyploid megakaryocytes, and specifically increases the number of platelets. The results of the TPO concentration determination in the serum and bone marrow supernatant of each group of mice are shown in Table 3. From the results in Table 3, it can be seen that the serum TPO and bone marrow supernatant TPO concentrations of the model group mice were significantly lower than those of the blank control group (P < 0.05), indicating that the mouse model was successfully established. The serum TPO concentration and bone marrow supernatant TPO concentration of mice in each medication group were significantly higher than those in the model control group (P < 0.01); the increase in serum TPO concentration and bone marrow supernatant TPO concentration of mice in the low, medium and high dose groups of the composition of the present invention was dose-dependent.

[0069] Table 3 Comparison of TPO concentrations in serum and bone marrow supernatant of mice in each group ( n=10)

[0070]

[0071]

[0072] Note: Compared with the blank control group, "*" indicates P < 0.05, "**" indicates P < 0.01; compared with the model control group, ▲ "P<0.05," ▲▲ ” indicates P<0.01.

[0073] 3.4 Determination of bone marrow megakaryocyte counts in each group of mice

[0074] The results of bone marrow megakaryocyte counts of mice in each group are shown in Table 4. As shown in Table 4, the bone marrow megakaryocyte counts of mice in the model control group were significantly lower than those in the blank control group (P < 0.01). The bone marrow megakaryocyte counts of mice in each drug administration group were significantly higher than those in the model control group (P < 0.01).

[0075] Table 4 Comparison of bone marrow megakaryocyte counts in each group of mice ( n=10)

[0076]

[0077] Note: Compared with the blank control group, "**" indicates P < 0.01; compared with the model control group, ▲▲ ” indicates P<0.01.

[0078] From the above results, it can be seen that the Chinese medicine composition of the present invention can promote the platelet count of mice with thrombocytopenia caused by cyclophosphamide, promote the production of megakaryocytes in the mouse bone marrow, and increase the TPO level.

[0079] Experimental Example 2 Effect of the Composition of the Present Invention on Carboplatin-Induced Thrombocytopenia in Mice

[0080] 1 Experimental Materials

[0081] Instruments and reagents: Recombinant human interleukin-11 for injection, specification: 3 mg (24 million U) / vial, produced by Qilu Pharmaceutical Co., Ltd.; Carboplatin injection, specification 10 ml: 50 mg, produced by Qilu Pharmaceutical Co., Ltd.; The composition of the present invention is the mixture prepared in Example 2. Mouse thrombopoietin ELISA kit, produced by Wuhan Boster Bioengineering Co., Ltd.; Sysmex-K21 blood cell analyzer, Jinan Sysmex Medical Electronics Co., Ltd.

[0082] Experimental animals: 60 Kunming mice, half male and half female, 6-8 weeks old, body weight 18-22 g, experimental animal license number SYXK (Lu) 20180008, provided by Lunan Pharmaceutical Group Co., Ltd., adaptive feeding for one week before the experiment, free access to food and water during the experiment, 12 h circadian rhythm.

[0083] 2 Experimental methods

[0084] 2.1 Animal grouping, drug administration and modeling

[0085] 60 Kunming mice were randomly divided into 6 groups, each with 10 mice, namely, a blank control group, a model control group, a positive control group, a low-dose group, a medium-dose group and a high-dose group of the composition of the present invention.

[0086] The blank control group and the model control group were administered with an equal volume of normal saline, the positive control group was subcutaneously injected with 5 μg / kg of recombinant human interleukin-11 solution, and the injection volume was 10 ml / kg. The low, medium and high dose groups of the composition of the present invention were intragastrically administered with the mixture prepared in Example 2, and 8.4 g of raw medicinal materials / kg, 16.7 g of raw medicinal materials / kg, and 32.3 g of raw medicinal materials / kg were administered respectively, and the administration was continued for 14 days, once a day.

[0087] Modeling method: On the 8th day of administration, except for the blank control group, the mice in other groups were injected with carboplatin solution once through the tail vein at a concentration of 5 mg / ml and an injection volume of 0.15 ml / 10 g body weight. The mice in the blank control group were injected with normal saline through the tail vein at the same dosage volume to establish a mouse model of thrombocytopenia induced by carboplatin chemotherapy. The standard for successful modeling was that the platelet count of the mice in the model control group decreased to 20%-30% of the platelet count of the mice in the blank control group.

[0088] 2.2 Index observation and measurement

[0089] 2.2.1 Mouse peripheral platelet count

[0090] Orbital venous blood was collected from mice before the start of drug administration (d0), after model establishment (d9), and 1 h after the last drug administration (d14), and the platelet count was determined using a Sysmex-K21 blood cell analyzer.

[0091] 2.2.2 Determination of megakaryocyte count in mouse femoral bone marrow

[0092] The mice were killed by cervical dislocation, and the femurs on both sides of the mice were freed. 1 ml of PBS buffer was drawn into a 1 ml syringe to flush out all the bone marrow cells in the femurs. The cell suspension was passed through a syringe with a No. 4 needle to fully disperse the cells in the suspension. After centrifugation and aspiration of the bone marrow supernatant, the bone marrow cell fluid was smeared and stained, and the number of all bone marrow megakaryocytes was counted under a microscope using the white blood cell counting method.

[0093] 2.3 Experimental data processing and analysis

[0094] The experimental data were statistically analyzed using SPSS22 software, and the data were expressed as mean ± standard deviation. The results were analyzed by one-way analysis of variance, and the t-test was used to compare the means of the two groups. P < 0.01 and P < 0.05 indicated that the difference between the two groups was statistically significant.

[0095] 3 Experimental results

[0096] 3.1 Comparison of PLT counts in each group of mice

[0097] The effect of the composition of the present invention on the PLT counts of each group of mice is shown in Table 5. From the results in Table 5, it can be seen that the platelet count of mice in the model control group on the 9th day was significantly higher than that of mice in the blank control group (P < 0.05). The platelet count of mice in each administration group on the 14th day was significantly higher than that of mice in the model control group (P < 0.05, P < 0.01). It shows that both the positive control drug and the composition of the present invention can increase the peripheral blood PLT count of mice with platelet reduction caused by carboplatin.

[0098] Table 5 Changes in peripheral blood PLT counts of mice in each group ( n=10)

[0099]

[0100] Note: Compared with the blank control group, "*" indicates P < 0.05; compared with the model control group, ▲ "P<0.05," ▲▲ ” indicates P<0.01.

[0101] 3.2 Determination of bone marrow megakaryocyte counts in each group of mice

[0102] The results of the determination of bone marrow megakaryocyte counts of mice in each group are shown in Table 6. As can be seen from Table 6, the bone marrow megakaryocyte counts of mice in the model control group were significantly lower than those in the blank control group (P < 0.01). The bone marrow megakaryocyte counts of mice in the positive control group and the medium and high dose groups of the composition of the present invention were significantly higher than those in the model control group (P < 0.05 (P < 0.01).

[0103] Table 6 Comparison of bone marrow megakaryocyte counts in each group of mice ( n=10)

[0104]

[0105]

[0106] Note: Compared with the blank control group, "**" indicates P < 0.01; compared with the model control group, ▲▲ ” indicates P<0.01.

[0107] It can be seen from the above test results that the Chinese medicine composition of the present invention can promote the platelet count of mice with thrombocytopenia caused by carboplatin and promote the production of megakaryocytes in the mouse bone marrow. DETAILED DESCRIPTION

[0108] The technical solution of the present invention is described below by way of examples. Parameters not described in detail in the examples may be explained by the manufacturer's suggestions or the instructions.

[0109] Example 1 Granule Preparation

[0110]

[0111] (1) Remove impurities from Scrophularia and Angelica, wash, soak, cut into thin slices, and set aside; remove impurities from Atractylodes, Sargentodoxae, and Radix Rehmanniae, wash, soak, cut into thick slices, dry, and set aside; remove impurities from Agrimonia, Taraxacum, and Eclipta prostrata, wash, cut into sections, dry, and set aside;

[0112] (2) taking the ten medicinal materials processed in step (1) in the prescription amount, adding water and decocting them twice, the first time adding 10 times the amount of water and decocting for 3 hours, the second time adding 8 times the amount of water and decocting for 2 hours; filtering, collecting and combining the decoctions, and concentrating under reduced pressure to an extract with a relative density of 1.32 at 50°C;

[0113] (3) The extract obtained in step (2) is vacuum dried to obtain a dry paste, and appropriate amounts of sucrose and starch are added, mixed, softened with 80% ethanol, granulated, and dried to obtain granules.

[0114] Example 2 Preparation of mixture

[0115]

[0116] (1) Remove impurities from Scrophularia and Angelica, wash, soak, cut into thin slices, and set aside; remove impurities from Atractylodes, Sargentodoxae, and Radix Rehmanniae, wash, soak, cut into thick slices, dry, and set aside; remove impurities from Agrimonia, Taraxacum, and Eclipta prostrata, wash, cut into sections, dry, and set aside;

[0117] (2) Take the ten medicinal materials processed in step (1) and add water to boil twice. The first time, add 10 times the amount of water and boil for 3 hours. The second time, add 8 times the amount of water and boil for 1.5 hours. Combine the decoctions and concentrate under reduced pressure to an extract with a relative density of 1.07 (55° C.);

[0118] (3) adding ethanol to the extract obtained in step (2) until the alcohol content is 65%, cooling and precipitating for 48 hours, filtering and obtaining the alcohol precipitate for later use;

[0119] (4) Recover ethanol by vacuum concentration and concentrate until the relative density of the concentrate is 1.26 (55°C), add 6 times the amount of water, cool and settle for 48 hours, filter, concentrate the supernatant to an appropriate amount, add an appropriate amount of aspartame, stir to dissolve, adjust the pH to 7.0, add water to the total amount of preparation, stir, filter until the clarity is acceptable, fill, and sterilize to obtain the mixture.

[0120] Example 3 Preparation of oral liquid

[0121]

[0122] (1) Remove impurities from Scrophularia and Angelica, wash, soak, cut into thin slices, and set aside; remove impurities from Atractylodes, Sargentodoxae, and Radix Rehmanniae, wash, soak, cut into thick slices, dry, and set aside; remove impurities from Agrimonia, Taraxacum, and Eclipta prostrata, wash, cut into sections, dry, and set aside;

[0123] (2) taking the ten medicinal materials processed in step (1) in the prescription amount, adding water and decocting them twice, the first time adding 8 times the amount of water and decocting for 2 hours, and the second time adding 6 times the amount of water and decocting for 1 hour; filtering, collecting and combining the decoctions, and concentrating under reduced pressure to an extract with a relative density of 1.06 at 60°C;

[0124] (3) adding ethanol to the extract obtained in step (2) until the alcohol content is 60%, cooling and precipitating for 48 hours, filtering and obtaining the alcohol precipitate for later use;

[0125] (4) Concentrate under reduced pressure to recover ethanol and concentrate until the relative density of the concentrate is 1.21 (60°C), add 6 times the amount of water, cool and settle for 48 hours, filter, centrifuge and filter the supernatant, add appropriate amount of sucralose, stir to dissolve, adjust pH to 7.0, add water to the total amount of preparation, stir, filter until the clarity is acceptable, fill and sterilize to obtain the oral solution.

[0126] Example 4 Preparation of capsules

[0127]

[0128] (1) Remove impurities from Scrophularia and Angelica, wash, soak, cut into thin slices, and set aside; remove impurities from Atractylodes, Sargentodoxae, and Radix Rehmanniae, wash, soak, cut into thick slices, dry, and set aside; remove impurities from Agrimonia, Taraxacum, and Eclipta prostrata, wash, cut into sections, dry, and set aside;

[0129] (2) taking the ten medicinal materials processed in step (1) in the prescription amount, adding water and decocting twice, the first time adding 8 times the amount of water and decocting for 2 hours, the second time adding 6 times the amount of water and decocting for 1 hour; filtering, collecting and combining the decoctions, and concentrating under reduced pressure to an extract with a relative density of 1.06 at 55°C;

[0130] (3) adding ethanol to the extract obtained in step (2) until the alcohol content is 60%, cooling and precipitating for 48 hours, filtering and obtaining the alcohol precipitate for later use;

[0131] (4) Concentrating under reduced pressure to recover ethanol and concentrating to a relative density of the concentrate of 1.35 (55° C.), vacuum drying to obtain a dry paste, crushing it into fine powder, sieving, mixing, and filling it into capsule shells to prepare capsules.

[0132] Example 5 Preparation of tablets

[0133]

[0134] (1) Remove impurities from Scrophularia and Angelica, wash, soak, cut into thin slices, and set aside; remove impurities from Atractylodes, Sargentodoxae, and Radix Rehmanniae, wash, soak, cut into thick slices, dry, and set aside; remove impurities from Agrimonia, Taraxacum, and Eclipta prostrata, wash, cut into sections, dry, and set aside;

[0135] (2) taking the ten medicinal materials processed in step (1) in the prescription amount, adding water and decocting them twice, the first time adding 12 times the amount of water and decocting for 2 hours, and the second time adding 8 times the amount of water and decocting for 1 hour; filtering, collecting and combining the decoctions, and concentrating under reduced pressure to an extract with a relative density of 1.38 at 50°C;

[0136] (3) The extract obtained in step (2) is vacuum dried to obtain a dry paste, which is crushed into fine powder, and appropriate amounts of microcrystalline cellulose and magnesium stearate are added, mixed, and compressed to obtain tablets.

[0137] Example 6 Preparation of Pills

[0138]

[0139] (1) Remove impurities from Scrophularia and Angelica, wash, soak, cut into thin slices, and set aside; remove impurities from Atractylodes, Sargentodoxae, and Radix Rehmanniae, wash, soak, cut into thick slices, dry, and set aside; remove impurities from Agrimonia, Taraxacum, and Eclipta prostrata, wash, cut into sections, dry, and set aside;

[0140] (2) taking the ten medicinal materials processed in step (1) in the prescription amount, adding water and decocting twice, the first time adding 12 times the amount of water and decocting for 2 hours, the second time adding 8 times the amount of water and decocting for 2 hours; filtering, collecting and combining the decoctions, and concentrating under reduced pressure to an extract with a relative density of 1.36 at 50°C;

[0141] (3) The extract obtained in step (2) is vacuum dried to obtain a dry paste, which is crushed, sieved, and then pelletized with 80% ethanol and dried to obtain pills.

Claims

1. A Chinese medicine composition for preventing and treating bone marrow suppression after chemotherapy, characterized in that: It is made from the following Chinese medicinal ingredients: Scrophularia, Atractylodes, Astragalus, Sargentodoxae, Rehmannia glutinosa, Radix Angelicae Sinensis, Herba Agrimoniae, Caulis Spatholobi, Taraxaci and Eclipta prostrata.

2. The Chinese medicine composition according to claim 1, characterized in that: The Chinese medicine composition is made of the following Chinese medicine components in parts by weight: Scrophularia 9-20 weight parts Atractylodes 6-15 weight parts Astragalus 9-30 weight parts Sargentodoxae 9-20 weight parts Rehmannia glutinosa 10-20 weight parts Angelica 6-15 weight parts Agrimoniae 6-15 weight parts Millettia reticulata 9-20 weight parts Taraxacum officinale 10-20 weight parts Eclipta prostrata 6-15 weight parts.

3. The Chinese medicine composition according to claim 2, characterized in that: The Chinese medicine composition is made of the following Chinese medicine components in parts by weight: Scrophularia 12-15 weight parts Atractylodes 10-12 weight parts Astragalus 15-20 weight parts Sargentodoxae 10-15 weight parts Rehmannia glutinosa 12-15 weight parts Angelica 10-12 weight parts Agrimonia 10-12 weight parts Millettia reticulata 10-15 weight parts Taraxacum officinale 12-15 weight parts Eclipta prostrata 10-12 weight parts.

4. The Chinese medicine composition according to claim 3, characterized in that: The Chinese medicine composition is made of the following Chinese medicine components in parts by weight: 15 parts by weight of Scrophularia ningpoensis, 10 parts by weight of Atractylodes macrocephala, 20 parts by weight of Astragalus membranaceus, 12 parts by weight of Sargentodoxae, 12 parts by weight of Rehmannia glutinosa, 12 parts by weight of Angelica sinensis, 10 parts by weight of Agrimoniae, 12 parts by weight of Millettia reticulata, 12 parts by weight of Taraxacum officinale, 15 parts by weight of Eclipta prostrata.

5. A method for preparing the Chinese medicine composition according to any one of claims 1 to 4, characterized in that: The steps include: (1) Remove impurities from Scrophularia and Angelica, wash, soak, cut into thin slices, and set aside; remove impurities from Atractylodes, Sargentodoxae, and Radix Rehmanniae, wash, soak, cut into thick slices, dry, and set aside; remove impurities from Agrimonia, Taraxacum, and Eclipta prostrata, wash, cut into sections, dry, and set aside; (2) taking 10 raw medicinal materials processed in step (1) in the prescribed amount, adding 6-12 times of water and boiling and extracting for 2-3 times, each time for 1-3 hours; filtering, collecting and combining the decoction, and concentrating under reduced pressure to a relative density of 1.05-1.4 (50-60° C.), to obtain an extract; (3) adding pharmaceutically acceptable excipients to the extract obtained in step (2) and preparing the extract into a clinically acceptable dosage form through conventional processes.

6. The preparation method according to claim 5, characterized in that: The clinically acceptable dosage form is one of granules, tablets, pills, capsules, oral liquids or mixtures.

7. Use of the Chinese medicine composition according to claim 1 in the preparation of drugs for preventing and treating bone marrow suppression after chemotherapy.

8. The use according to claim 7, characterized in that: The bone marrow suppression is manifested as a decrease in white blood cells, red blood cells and / or platelets after chemotherapy.

9. The use according to claim 7, characterized in that: The composition is used in preparing a drug for preventing and treating thrombocytopenia after chemotherapy.

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