Traditional chinese medicine composition for promoting peripheral platelet production and preparation thereof

Abstract

The present invention relates to a traditional Chinese medicine composition for promoting peripheral platelet production and a preparation thereof. The traditional Chinese medicine composition is prepared from Loropetalum chinense or an extract thereof and Scutellariae radix or an extract thereof, wherein the traditional Chinese medicine composition comprises the following components: 1 to 10 parts by weight of Loropetalum chinense or the extract thereof, and 1 to 10 parts by weight of Scutellariae radix or the extract thereof. The traditional Chinese medicine composition can significantly increase peripheral blood platelet level in a thrombocytopenia model animal, promote peripheral platelet production, and is suitable for the treatment of thrombocytopenia.

Description

A traditional Chinese medicine composition for promoting peripheral thrombopoiesis and a preparation thereof TECHNICAL FIELD

[0001] The present application belongs to the field of traditional Chinese medicine extracts, and particularly relates to a traditional Chinese medicine composition for promoting peripheral thrombopoiesis and a preparation thereof. BACKGROUND

[0002] Loropetalum chinense, also known as Loropetalum, is a plant of Loropetalum in Hamamelidaceae. Loropetalum chinense (R. Br.) Oliver and its variety Loropetalum chinense var. rubrum are also plants of Loropetalum in Hamamelidaceae. Loropetalum is usually a shrub or small tree, and is widely distributed in central, southern and southwestern provinces, and also has rich wild resources in China. The flowers, leaves and roots of Loropetalum can be used as medicine, and have the effects of clearing heat and detoxifying, astringing, hemostasis, etc., and are commonly used for the treatment of burns, scalds, external injuries, hemorrhage, metrorrhagia, diarrhea, gastric ulcer, etc. At present, more than 60 compounds have been isolated from Loropetalum, including tannins, flavonoid glycosides, lignans, steroids, etc., which have the effects of antibacterial, anti-inflammatory, antioxidant, promoting healing, etc., and have high clinical application value and broad market prospects.

[0003] Scutellaria baicalensis Georgi, also known as Huangqin, is a dry root of Scutellaria baicalensis Georgi in Labiatae. As a medicine, it is recorded in Shennong Bencao Jing, and Chinese Pharmacopoeia 2020 records that it tastes bitter and is cold in nature; it is related to the lung, gallbladder, spleen, small intestine and large intestine channels, and has the effects of clearing heat and drying dampness, purging fire and detoxifying, stopping bleeding, and preventing miscarriage. Modern studies have shown that Scutellaria baicalensis Georgi contains flavonoids, organic acids, volatile oils, polysaccharides and other components, and has the effects of anti-tumor, anti-inflammatory, antibacterial, antioxidant, etc.

[0004] Thrombocytopenia is a refractory hematological disease, mainly including primary immune thrombocytopenia, secondary thrombocytopenia caused by diseases such as malignant tumors, systemic lupus erythematosus, chronic anemia and drug-induced thrombocytopenia caused by chemotherapy drugs, thrombolytic drugs, etc. The incidence of thrombocytopenia in China is high (5-10 per 100,000 people), and patients generally have symptoms such as skin purpura, fatigue, anxiety, and may be accompanied by leukopenia, anemia and total hemoglobin reduction, and severe cases can die due to internal organ hemorrhage or intracranial hemorrhage. The etiology of thrombocytopenia is complex, and the pathogenesis has not been fully elucidated, and there is a lack of effective clinical treatment drugs. Glucocorticoids and gamma globulin are the first-line treatment drugs for thrombocytopenia, but there are problems such as low efficiency, large side effects, high cost, inability to take long-term medication, easy recurrence after stopping medication, etc. Human recombinant thrombopoietin injection (such as Tiberio, etc.), oral thrombopoietin receptor agonists (such as eltrombopag, romiplostim, etc.) and rituximab are also widely used for various types of primary and secondary thrombocytopenia. Although such drugs can be taken long-term, the clinical efficiency is only about 60%, and there are also problems such as high treatment cost, large side effects, easy drug resistance and easy recurrence after stopping medication.

[0005] Domestic research on E. indica and E. acida mainly focuses on the isolation of compounds and medicinal activity. Zhang Wugang et al. also recorded that E. indica leaves have the effects of "relieving blood stasis, promoting blood circulation, relieving fever and diarrhea, treating abdominal pain, muscle stiffness, diarrhea, callus and external trauma bleeding" (Analysis of Hemostatic Chemical Constituents of E. indica, Chinese Journal of Experimental Traditional Chinese Formulas, 2017, 23(05): 47-52) and other effects. Lian Zeqin et al. reported that E. indica can promote the repair of damaged epithelial tissue and promote wound healing, and its hemostatic and coagulation effects are significant and accurate (Preliminary Study on the Material Basis of E. indica for Promoting Wound Healing in Rats, Chinese Journal of Chinese Materia Medica, 2013, 38(20): 3566-3570). It is also widely reported that E. indica is clinically used for treating external trauma bleeding, postpartum uterine contraction and bleeding, and promoting tissue healing at the surgical site.

[0006] Although E. indica and E. acida have excellent hemostatic and anti-inflammatory effects, unlike the mechanism of traditional Chinese medicine hemostasis, which mainly promotes blood vessel contraction, coagulation factor release and platelet aggregation, the treatment of thrombocytopenia is mainly through promoting the maturation of megakaryocytes in bone marrow, the generation of platelets and the reduction of megakaryocyte and platelet destruction. There is no report on the promotion of peripheral platelet generation by E. indica extract and E. acida extract and their combination. SUMMARY

[0007] The present application aims to provide a traditional Chinese medicine composition, which is characterized in that it is made of the following weight parts of traditional Chinese medicine raw materials: 1-10 parts by weight of E. indica or its extract, 1-10 parts by weight of E. acida or its extract.

[0008] wherein, is made of the following weight parts of traditional Chinese medicine raw materials: 1-9 parts by weight of Rhododendron simsii or extract thereof, 1-10 parts by weight of Scutellaria baicalensis or extract thereof;

[0009] or, 1-7 parts by weight of Rhododendron simsii or extract thereof, 3-10 parts by weight of Scutellaria baicalensis or extract thereof;

[0010] or, 5-9 parts by weight of Rhododendron simsii or extract thereof, 5-10 parts by weight of Scutellaria baicalensis or extract thereof;

[0011] or, 7-9 parts by weight of Rhododendron simsii or extract thereof, 3-5 parts by weight of Scutellaria baicalensis or extract thereof;

[0012] or, 1-9 parts by weight of Rhododendron simsii or extract thereof, 1-9 parts by weight of Scutellaria baicalensis or extract thereof.

[0013] Preferably, is made of the following weight parts of traditional Chinese medicine raw materials: 9 parts by weight of Rhododendron simsii or extract thereof, 1 part by weight of Scutellaria baicalensis or extract thereof;

[0014] or, 7 parts by weight of Rhododendron simsii or extract thereof, 3 parts by weight of Scutellaria baicalensis or extract thereof;

[0015] or, 5 parts by weight of Rhododendron simsii or extract thereof, 5 parts by weight of Scutellaria baicalensis or extract thereof;

[0016] or, 3 parts by weight of Rhododendron simsii or extract thereof, 7 parts by weight of Scutellaria baicalensis or extract thereof;

[0017] or, 1 part by weight of Rhododendron simsii or extract thereof, 9 parts by weight of Scutellaria baicalensis or extract thereof.

[0018] wherein, the Rhododendron simsii extract is prepared by the following method: Rhododendron simsii is crushed, sieved, an alcohol organic solvent is added as an extractant, reflux extraction is performed, the extract is combined, and the extract is concentrated under reduced pressure to obtain an extract, the extract is adsorbed by a macroporous resin column, deionized water is used for elution, and then different concentrations of alcohol organic solvents are used for elution, different concentrations of eluate are collected, and the Rhododendron simsii extract is obtained by concentration under reduced pressure;

[0019] the Scutellaria baicalensis extract is prepared by the following method: Scutellaria baicalensis is crushed, sieved, an alcohol organic solvent is added as an extractant, reflux extraction is performed, the extract is combined, and the extract is concentrated under reduced pressure to obtain an extract, the extract is adsorbed by a macroporous resin column, deionized water is used for elution, and then different concentrations of alcohol organic solvents are used for elution, different concentrations of eluate are collected, and the Rhododendron simsii extract is obtained by concentration under reduced pressure;

[0020] the macroporous resin column is selected from one of the following types: HPD-300, HPD-400, HPD-450, HPD-600, HPD-826, ADS-17, ADS-21, AB-8, D101, or D301.

[0021] Wherein, in the preparation of the Loropetalum chinensis extract or the Scutellaria baicalensis extract, the material-liquid ratio of Loropetalum chinensis or Scutellaria baicalensis to the extractant is 1:1-12 kg / L, reflux extraction is performed 1 to 4 times, and each extraction time is 0.2 to 4 hours. The alcohol organic solvent is methanol, ethanol, propanol or butanol; the mass fraction of the alcohol organic solvent is 5%-100%, 10%-80%, 20%-60% or 40%-60%;

[0022] Preferably, Loropetalum chinensis or Scutellaria baicalensis is sieved through 18 mesh; the material-liquid ratio of Loropetalum chinensis or Scutellaria baicalensis to the extractant is 1:2-10 kg / L, reflux extraction is performed 1-3 times, each extraction time is 0.5-2 hours, and the extract is concentrated under reduced pressure to a relative density of 1.10-1.30 (60°C); the alcohol organic solvent is ethanol; the mass fraction of the alcohol organic solvent is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 100%, and the dosage is 2-8 BV.

[0023] In the preparation of the Loropetalum chinensis extract or the Scutellaria baicalensis extract, the macroporous resin column is selected from one of the following models: HPD-300, HPD-400, ADS-17 or D101, and the adsorption flow rate is 1-8 mL·min -1 The diameter of the macroporous resin is 1-6 cm, the diameter-to-height ratio is 1:2-8, the sample volume is 10-60 mL, and the static adsorption is 2-24 h; the amount of ionized water used for elution is 1 BV-6 BV,

[0024] Preferably, the adsorption flow rate is 2-6 mL·min -1 The diameter of the macroporous resin is 2-4 cm, the diameter-to-height ratio is 1:2-4, the sample load is 20-40 mL, and the static adsorption is 4-24 h; the amount of ionized water used for elution is 2-6 BV.

[0025] Wherein, the loropetalum is white loropetalum or red loropetalum.

[0026] The present invention aims to provide a method for preparing a Chinese medicine composition, which is characterized in that the preparation method is as follows:

[0027] (1) Preparation of Loropetalum chinense extract: The Loropetalum chinense was crushed and sieved, and an alcoholic organic solvent was added as an extractant. The extracts were refluxed and concentrated under reduced pressure to obtain an extract. The extract was adsorbed on a macroporous resin column, eluted with deionized water, discarded, and then eluted with alcoholic organic solvents of different concentrations. The eluates of different concentrations were collected and concentrated under reduced pressure to obtain the Loropetalum chinense extract.

[0028] (2) Scutellaria extract preparation: Scutellaria is crushed, sieved, alcohol organic solvent is added as extractant, reflux extraction, combined extract, reduced pressure concentration to obtain the extract, the extract is adsorbed by macroporous resin column, deionized water elution, discarded, and then eluted with different concentrations of alcohol organic solvent, and the eluate of different concentrations is collected and concentrated under reduced pressure to obtain the extract of wampee;

[0029] The macroporous resin column is selected from one of the following types: HPD-300, HPD-400, HPD-450, HPD-600, HPD-826, ADS-17, ADS-21, AB-8, D101 or D301;

[0030] (3) The wampee extract and the Scutellaria extract are mixed in a ratio of 1-10:1-10 by weight.

[0031] In the preparation of the wampee extract in step (1) or the Scutellaria extract in step (2), the ratio of wampee or Scutellaria to the extractant is 1:1-12 kg / L, the reflux extraction is performed 1-4 times, and the extraction time is 0.2-4 h each time; the alcohol organic solvent is methanol, ethanol, propanol or butanol; and the mass fraction of the alcohol organic solvent is 5%-100%, 10%-80%, 20%-60% or 40%-60%.

[0032] Preferably, the wampee or Scutellaria is sieved to 18 mesh; the ratio of wampee or Scutellaria to the extractant is 1:2-10 kg / L, the reflux extraction is performed 1-3 times, and the extraction time is 0.5-2 h each time; the extract is concentrated under reduced pressure to a relative density of 1.10-1.30 (60°C); the alcohol organic solvent is ethanol; the mass fraction of the alcohol organic solvent is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 100%; and the amount of the alcohol organic solvent is 2-8 BV.

[0033] More preferably, the macroporous resin column is selected from one of the following types: HPD-300, HPD-400, ADS-17 or D101, the adsorption flow rate is 1-8 mL·min -1 , the diameter of the macroporous resin is 1-6 cm, the diameter-height ratio is 1:2-8, the loading amount is 10-60 mL, the static adsorption time is 2-24 h, and the deionized water elution amount is 1 BV-6 BV.

[0034] Further preferably, the adsorption flow rate is 2-6 mL·min -1 , the diameter of the macroporous resin is 2-4 cm, the diameter-height ratio is 1:2-4, the loading amount is 20-40 mL, the static adsorption time is 4-24 h, and the deionized water elution amount is 2-6 BV.

[0035] The extract of Loropetalum chinense and the extract of Scutellaria baicalensis in the step (3) are in a weight ratio of 9:1, 7:3, 5:5, 3:7 or 1:9.

[0036] The composition is in a form of a pharmaceutical preparation, and the pharmaceutical preparation can further comprise a pharmaceutical excipient as needed.

[0037] The pharmaceutical preparation is selected from the group consisting of a tablet, a capsule, a buccal, a granule, a pill, a powder, a paste, a bolus, a suspension, a solution, an injection, a suppository, a plaster, a spray, a drop, a drop pill or a patch.

[0038] The application aims to provide the use of the traditional Chinese medicine composition in the preparation of a medicine for blood system diseases.

[0039] The blood system disease includes but is not limited to iron deficiency anemia, aplastic anemia, acute leukemia, idiopathic thrombocytopenic purpura (ITP), primary immune thrombocytopenia, chronic anemia or thrombocytopenia, preferably thrombocytopenia.

[0040] The medicine can promote peripheral platelet production, and improve the levels of peripheral red blood cells and hemoglobin.

[0041] The following is the explanation and description of the terms of the application:

[0042] Obviously, according to the above content of the application, according to the ordinary technical knowledge and common means in the art, other modifications, replacements or changes can be made without departing from the above basic technical idea of the application.

[0043] The information related to the prescription of the prescription medicine is as follows:

[0044] Scutellaria baicalensis: the product is the dried root of Scutellaria baicalensis Georgi of the Labiatae family. The product should meet the relevant provisions of Scutellaria baicalensis in Chinese Pharmacopoeia 2020 edition part 1 page 314.

[0045] The Loropetalum chinense in the application includes Loropetalum chinense or Loropetalum chinense.

[0046] The term "pharmaceutically acceptable" means that a carrier, vehicle, diluent, excipient and / or formed salt is generally chemically or physically compatible with other ingredients constituting a pharmaceutical dosage form, and is physiologically compatible with the recipient.

[0047] The pharmaceutically acceptable carrier includes diluents, excipients, fillers, emulsifiers, binders, lubricants, absorption promoters, surfactants, disintegrants or antioxidants that are conventional in the pharmaceutical field. The pharmaceutically acceptable carrier also includes flavoring agents, sweetening agents, preservatives or coloring agents. For example: sorbitol, sodium pyrosulfite, sodium bisulfite, sodium thiosulfate, soybean phospholipid, vitamin C, vitamin E, disodium EDTA, carbonates of monovalent alkali metals, acetates, phosphates or aqueous solutions thereof, hydrochloric acid, acetic acid, sulfuric acid, phosphoric acid, amino acids, sodium chloride, potassium chloride, sodium lactate, hydroxyethyl benzoate solution, benzoic acid, potassium sorbate, chlorhexidine acetate, maltose, glucose, dextran, starch, sucrose, lactose, mannitol, cellulose and its derivatives, alginate, gelatin, polyvinylpyrrolidone, glycerol, Tween 80, agar, calcium carbonate, calcium bicarbonate, surfactants, polyethylene glycol, cyclodextrin, phospholipid materials, kaolin, talc, calcium stearate or magnesium stearate, etc.

[0048] The traditional Chinese medicine preparation of the present application can be an oral solid preparation, an oral liquid preparation or an injection; wherein the oral solid preparation can be a capsule in addition to tablets; the oral liquid preparation is selected from aqueous or oily suspensions, solutions, emulsions, syrup; the injection is selected from emulsion, lyophilized powder injection and water injection.

[0049] The dosage of the extract and its effective ingredients of the present application depends on the age, weight and type and severity of the disease of the subject.

[0050] The beneficial technical effects of the present application are: 1. The extract of Loropetalum chinense and the extract of Scutellaria baicalensis are combined into effective ingredients, the ratio is clear, which can ensure the stability of drug quality and the uniformity of drug efficacy, and can be further developed into a traditional Chinese medicine preparation suitable for clinical application. Compared with the prior art, the preparation method of the traditional Chinese medicine composition provided by the present application is simple and suitable for industrial production.

[0051] 2. The present application provides the application of the traditional Chinese medicine extract in promoting peripheral platelet production, the preparation can promote the increase of peripheral platelet count in the guinea pig anti-platelet serum to immunogenic platelet reduction syndrome mouse model, and improve the peripheral red blood cell and hemoglobin count level. DETAILED DESCRIPTION

[0052] The present application is further illustrated by the following examples, but is not limited thereto.

[0053] Example 1 Preparation of traditional Chinese medicine extract A

[0054] 1. The composition and formula of the traditional Chinese medicine extract A are: Radermachia hainanensis extract and Scutellaria baicalensis extract. The Radermachia hainanensis extract and the Scutellaria baicalensis extract are mixed in a mass ratio of 10:0, 9:1, 7:3, 5:5, 3:7, 1:9, or 0:10 to obtain the traditional Chinese medicine extract A, wherein the preparation steps of the Radermachia hainanensis extract and the Scutellaria baicalensis extract are as shown below.

[0055] 2. The preparation method of the Radermachia hainanensis extract is as follows: the fresh Radermachia hainanensis material is crushed, then passed through an 18-mesh sieve, then 60% ethanol is added to the powder to obtain a material-liquid ratio of 1:8 kg / L, then the extract is obtained by condensation reflux extraction for 3 times, 2 h each time, the extract is combined and concentrated under reduced pressure, and the extract is concentrated under reduced pressure to a relative density of 1.10-1.30 (60°C). HPD-300, HPD-400, D101, and ADS-17 macroporous adsorption resins are selected for purification; first, the extract is adsorbed on the macroporous resin column at a flow rate of 5 ml / min, the diameter of the macroporous adsorption resin column is 4 cm, the diameter-height ratio is 1:4, the loading amount is 40 ml, and the static adsorption time is 4 h; then, 2 BV of deionized water is used for elution, then 5 BV of 10% ethanol, 6 BV of 30% ethanol, 6 BV of 50% ethanol, and 5 BV of 70% ethanol are used for elution, respectively, and the ethanol eluate is collected, concentrated and dried to obtain the Radermachia hainanensis extract component.

[0056] 3. The preparation method of the Scutellaria baicalensis extract is as follows: the Scutellaria baicalensis root is crushed, then passed through an 18-mesh sieve, then 60% ethanol is added to obtain a material-liquid ratio of 1:10 kg / L, then the extract is obtained by condensation reflux extraction for 2 times, 2 h each time, the extract is combined, and the extract is concentrated under reduced pressure to a relative density of 1.10-1.30 (60°C). HPD-300, HPD-400, and D101 macroporous adsorption resins are selected for purification; first, the extract is adsorbed on the macroporous resin column at a flow rate of 5 ml / min, the diameter of the macroporous adsorption resin column is 4 cm, the diameter-height ratio is 1:2, the loading amount is 30 ml, and the static adsorption time is 24 h. Then, 5 BV of deionized water is used for elution, then 8 BV of 70% ethanol is used for elution, the 70% ethanol eluate is collected, concentrated and dried to obtain the Scutellaria baicalensis extract component.

[0057] Preparation of the traditional Chinese medicine extract B in Example 2

[0058] 1. The composition and formula of the traditional Chinese medicine extract B are: Radermachia rubiginosa extract and Scutellaria baicalensis extract. The Radermachia rubiginosa extract and the Scutellaria baicalensis extract are mixed in a mass ratio of 10:0, 9:1, 7:3, 5:5, 3:7, 1:9, or 0:10 to obtain the composition B.

[0059] 2. Preparation method of Lagerstroemia speciosa extract: Fresh Lagerstroemia speciosa material is crushed, then passed through an 18-mesh sieve, then 60% ethanol is added to the powder, so that the material-to-liquid ratio is 1:8 kg / L, then the extract is obtained by condensation reflux extraction for 3 times, 2 h each time, the extract is combined, and the extract is concentrated under reduced pressure to a relative density of 1.10-1.30 (60°C). HPD-300, HPD-400, D101, and ADS-17 macroporous adsorption resins are selected for purification; first, the extract is adsorbed on the macroporous resin column at a flow rate of 5 ml / min, the diameter of the macroporous adsorption resin column is 2 cm, the diameter-to-height ratio is 1:2, the loading amount is 20 ml, and the static adsorption time is 24 h. Then, 2 BV of deionized water is used for elution, then 5 BV of 10% ethanol, 6 BV of 30% ethanol, 6 BV of 50% ethanol, and 5 BV of 70% ethanol are used for elution, respectively, and the ethanol eluate is collected, concentrated, and dried to obtain the Lagerstroemia speciosa extract component.

[0060] 3. Preparation method of Scutellaria baicalensis extract: Scutellaria baicalensis roots are crushed, then passed through an 18-mesh sieve, 60% ethanol is added, so that the material-to-liquid ratio is 1:10 kg / L, then the extract is obtained by condensation reflux extraction for 2 times, 2 h each time, the extract is combined, and the extract is concentrated under reduced pressure to a relative density of 1.10-1.30 (60°C). HPD-300, HPD-400, and D101 macroporous adsorption resins are selected for purification. First, the extract is adsorbed on the macroporous resin column at a flow rate of 5 ml / min, the diameter of the macroporous adsorption resin column is 2 cm, the diameter-to-height ratio is 1:4, the loading amount is 30 ml, and the static adsorption time is 2 h. Then, 5 BV of deionized water is used for elution, then 8 BV of 70% ethanol is used for elution, the 70% ethanol eluate is collected, concentrated, and dried to obtain the Scutellaria baicalensis extract component.

[0061] Example 3 Preparation of oral tablets

[0062] 100 g of any formula of the traditional Chinese medicine composition A in Example 1, 10 g of liquid paraffin, 25 g of talc, 30 g of starch, and 25 g of tartaric acid are prepared into 1000 tablets by tabletting.

[0063] Example 4 Preparation of oral tablets

[0064] 100 g of any formula of the traditional Chinese medicine composition B in Example 2, 10 g of liquid paraffin, 25 g of talc, 30 g of starch, and 25 g of tartaric acid are prepared into 1000 tablets by tabletting.

[0065] Example 5 Preparation of oral emulsion

[0066] 100 g of any formula of the traditional Chinese medicine composition A in Example 1, 30 g of soybean phospholipid or injectable soybean phospholipid, and 10 g of sodium oleate are taken, and distilled water is added to 100 ml to prepare 100 ml of traditional Chinese medicine composition oral emulsion.

[0067] Example 6 Preparation of oral emulsion

[0068] Take 100 g of any formula of the traditional Chinese medicine composition B in Example 2, 30 g of soybean phospholipid or injectable soybean phospholipid, 10 g of sodium oleate, and distilled water to 100 ml to prepare 100 ml of traditional Chinese medicine composition oral emulsion.

[0069] Example 7 Preparation of capsules

[0070] The capsules of this example are composed of a drug solution and a gel solution. Take 100 g of any formula of the traditional Chinese medicine composition A in Example 1, add 10 g of vitamin E as an antioxidant and 10 g of Tween 80 as an emulsifier to prepare a drug solution. Take gelatin, purified water, glycerol, and preservatives in a mass ratio of 1:2.5:1:5, add glycerol, purified water, and preservatives into a gel-making tank in turn, heat to 80°C, then add gelatin while continuously stirring and vacuumizing until the gelatin is completely dissolved, filter the gel solution, and store the filtrate at 60°C.

[0071] Example 8 Preparation of capsules

[0072] The capsules of this example are composed of a drug solution and a gel solution. Take 100 g of any formula of the traditional Chinese medicine composition B in Example 2, add 10 g of vitamin E as an antioxidant and 10 g of Tween 80 as an emulsifier to prepare a drug solution. Take gelatin, purified water, glycerol, and preservatives in a mass ratio of 1:2.5:1:5, add glycerol, purified water, and preservatives into a gel-making tank in turn, heat to 80°C, then add gelatin while continuously stirring and vacuumizing until the gelatin is completely dissolved, filter the gel solution, and store the filtrate at 60°C.

[0073] Effect Example 1 Traditional Chinese medicine composition tablets enhance immunogenicity in a thrombocytopenia model peripheral platelet count

[0074] A mouse model of thrombocytopenia was established by intraperitoneal injection of guinea pig anti-platelet serum, and the mice were orally administered with tablets of traditional Chinese medicine compositions A and B, respectively. It was found that the tablets of each composition significantly increased the platelet count of the mice, and the efficacy was significantly different from that of the model group. Moreover, the platelet-increasing effect of some composition tablets was comparable to that of the positive control drug eltrombopag.

[0075] Test purpose: To observe the effect of oral traditional Chinese medicine extract on peripheral blood in immune thrombocytopenia, and to prove its role in promoting platelet production.

[0076] Test substance: Each traditional Chinese medicine extract was prepared into an oral solution with a concentration of 500 mg / kg using distilled water.

[0077] Positive control drug: Eltrombopag oral tablets were prepared into an oral solution with a concentration of 50 mg / kg using distilled water.

[0078] Test method: BAL / BC mice, half male and half female. Randomly divided according to body weight: normal control group, model group, positive control group, composition A oral administration group, composition B oral administration group, composition C oral administration group, composition D oral administration group, composition E oral administration group, composition F oral administration group. A total of 9 groups, each group of 8 animals. According to the platelet separation kit, the platelets of BAL / BC mice were separated, and then the platelets were suspended in 0.01 mol / L PBS with pH 7.4, then the platelets were counted, and finally the number of platelets was adjusted to 1-2×109 / L. Take the prepared platelets, add equal amount of Freund's incomplete adjuvant or Freund's incomplete adjuvant, ultrasonic mixing for more than 60 min to form water-in-oil emulsion, that is, Freund's incomplete adjuvant or Freund's incomplete adjuvant antigen. A small amount of prepared adjuvant antigen was sucked into cold water to maintain its integrity, and when it became oil droplets, the required water-in-oil emulsion was obtained. Take the Freund's complete adjuvant antigen and inject it subcutaneously on the back, palm, abdomen and other parts of the guinea pig at 0 weeks, not less than 5 points / time, each point not less than 0.1 mL; Take the Freund's incomplete adjuvant antigen and inject it subcutaneously on the back, palm and abdomen and other parts of the guinea pig at 1, 2 and 4 weeks, not less than 5 points / time, each point not less than 0.1 mL, and disinfect the local area after the operation. At the 5th week, the mice were anesthetized again, and the heart blood was taken, and after standing for 30-60 min, the supernatant was obtained by centrifugation at 3000 rpm for 10 min, which was the guinea pig anti-platelet serum, which was stored in the refrigerator at-20℃ for standby. Before each modeling, the guinea pig anti-platelet serum was taken out from the refrigerator and incubated in a 56℃ water bath for 30 min for complement inactivation, then 5% BALB / c mouse red blood cell suspension was added to the guinea pig anti-platelet serum, and incubated in a 37℃ water bath for 1 h, then centrifuged at 3500 r / min for 10 min, and the supernatant was reserved. The guinea pig anti-platelet serum was adsorbed twice by the same method, and the guinea pig anti-platelet serum was diluted with physiological saline at a volume ratio of 40:1 for standby. The diluted guinea pig anti-platelet serum was injected into the abdominal cavity of the mice at a concentration of 5 μL / g, once every other day, and the total modeling period was 21 days. After the operation, the local area was disinfected. Each group was given corresponding drugs by gavage according to body weight, 0.1 ml / 10g, and the drug administration started after modeling, once a day, for 21 consecutive days, and the positive control group was given Eltrombopag 25 mg / kg orally every day. On the 21st day after modeling, 200 μL of blood was collected submandibularly, and the platelet index changes were detected using a hemocytometer.

[0079] Test results: On the 21st day of administration, the peripheral platelet level of the model group mice was significantly decreased compared with that of the normal mice, and each traditional Chinese medicine composition tablet could significantly promote the generation of peripheral platelets, and the efficacy of some composition tablets was equivalent to that of the positive control drug Eltrombopag, and the results are shown in Table 1.

[0080] Table 1 Peripheral platelet count results of different traditional Chinese medicine preparations in guinea pig anti-platelet serum-induced thrombocytopenia model mice on the 21st day

[0081] In the table, "-" represents that the platelet level of the treatment group is reduced or increased by less than 10% compared with the model control group; "+" represents that the platelet level of the treatment group is increased by 10% to 49% compared with the model control group; "++" represents that the platelet level of the treatment group is increased by 50 to 89% compared with the model control group; "+++" represents that the platelet level of the treatment group is increased by 90 to 129% compared with the model control group; and "++++" represents that the platelet level of the treatment group is increased by more than 130% compared with the model control group.

[0082] The platelet level of the treatment group compared with the model control group has a statistical difference by Student's-t test: *, p<0.05; **, p<0.01; ***, p<0.001. The platelet level of the treatment group compared with the composition A tablet (Radermachia hainanensis: Scutellaria baicalensis = 10:0) corresponding group has a statistical difference by Student's-t test: #, p<0.05; ##, p<0.01; ###, p<0.001. The platelet level of the treatment group compared with the composition A tablet (Radermachia hainanensis: Scutellaria baicalensis = 0:10) corresponding group has a statistical difference by Student's-t test: $, p<0.05; $$, p<0.01; $$$, p<0.001. The platelet level of the treatment group compared with the composition B tablet (Radermachia sinica: Scutellaria baicalensis = 10:0) corresponding group has a statistical difference by Student's-t test: &, p<0.05; &&, p<0.01; &&&, p<0.001. The platelet level of the treatment group compared with the composition B tablet (Radermachia sinica: Scutellaria baicalensis = 0:10) corresponding group has a statistical difference by Student's-t test: @, p<0.05; @@, p<0.01; @@@, p<0.001.

[0083] Conclusion: The prepared traditional Chinese medicine composition tablet has the effect of significantly improving the peripheral platelet count of the immunogenic thrombocytopenia model mice. The test results show that when the HPD300 macroporous adsorption resin is selected for purification, the 30% ethanol elution fraction of Radermachia hainanensis is collected, and the mass ratio of Radermachia hainanensis to Scutellaria baicalensis is 7:3, the prepared tablet has the optimal effect of improving the peripheral platelet count of the immunogenic thrombocytopenia model mice, and the effect is equivalent to that of the positive control drug eltrombopag.

[0084] Effect of the traditional Chinese medicine composition tablet in Example 2 on improving the peripheral red blood cell and hemoglobin levels of the immunogenic thrombocytopenia model

[0085] The mouse model of thrombocytopenia is induced by intraperitoneal injection of guinea pig anti-platelet serum, and the preferred Chinese medicine compositions A and B in Example 3 are orally administered in the form of tablets, and it is found that the Chinese medicine composition tablets prepared in the application have better effects than the positive control drug eltrombopag.

[0086] Test purpose: to observe the effects of oral Chinese medicine composition tablets on peripheral blood of immune thrombocytopenia, so as to prove that the Chinese medicine composition tablets have the effect of promoting platelet production.

[0087] Test object: the preferred Chinese medicine composition tablets are prepared into oral solutions with a concentration of 500 mg / kg by using distilled water.

[0088] Positive control drug: eltrombopag oral tablets are prepared into oral solutions with a concentration of 50 mg / kg by using distilled water.

[0089] Test method: BAL / BC mice, half male and half female. According to the body weight, they were randomly divided into: normal control group, model group, positive control group, composition A oral administration group, composition B oral administration group, composition C oral administration group, composition D oral administration group, composition E oral administration group, composition F oral administration group. A total of 9 groups, 8 animals in each group. According to the platelet separation kit, the platelets of BAL / BC mice were separated, and then the platelets were suspended in 0.01 mol / L PBS with pH 7.4, and then the platelets were counted, and finally the number of platelets was adjusted to 1-2×109 / L. Take the prepared platelets, add equal amount of Freund's incomplete adjuvant or Freund's incomplete adjuvant, ultrasonic mixing for more than 60 min to form water-in-oil emulsion, that is, Freund's incomplete adjuvant or Freund's incomplete adjuvant antigen. A small amount of prepared adjuvant antigen was sucked into cold water to maintain integrity, and when it became oil droplets, the required water-in-oil emulsion was obtained. Take the Freund's complete adjuvant antigen and inject it subcutaneously on the back, palm, abdomen and other parts of the guinea pig at 0 weeks, not less than 5 points / time, each point not less than 0.1 mL; Take the Freund's incomplete adjuvant antigen and inject it subcutaneously on the back, palm and abdomen and other parts of the guinea pig at 1, 2 and 4 weeks, not less than 5 points / time, each point not less than 0.1 mL, and disinfect the local area after the operation. At the 5th week, the mice were anesthetized again, and the heart blood was taken, and after standing for 30-60 min, the supernatant was obtained by centrifugation at 3000 rpm for 10 min, which was the guinea pig anti-platelet serum, which was stored in the refrigerator at-20℃ for standby. Before each modeling, the guinea pig anti-platelet serum was taken out from the refrigerator and incubated in a 56℃ water bath for 30 min for complement inactivation, then 5% BALB / c mouse red blood cell suspension was added to the guinea pig anti-platelet serum, and incubated in a 37℃ water bath for 1 h, then centrifuged at 3500 r / min for 10 min, and the supernatant was reserved. The guinea pig anti-platelet serum was adsorbed twice by the same method, and the guinea pig anti-platelet serum was diluted with physiological saline at a volume ratio of 40:1 for standby. The diluted guinea pig anti-platelet serum was injected into the abdominal cavity of the mice at a concentration of 5 μL / g, once every other day, and the total modeling period was 21 days. After the operation, the local area was disinfected. Each group was given corresponding drugs by gavage at 0.1 ml / 10g, and the modeling was started immediately after the modeling, and the drug was given once a day, for 21 days. The positive control group was given Eltrombopag 25 mg / kg orally every day. On the 21st day after modeling, 200 μL of blood was collected submandibularly, and the changes of red blood cells and hemoglobin were detected by a hemocytometer.

[0090] Test results: After 21 days of modeling, the peripheral red blood cells and hemoglobin levels of the model group mice decreased to a certain extent compared with the normal mice. At this time, the positive control drug Eltrombopag oral tablet had no obvious effect on the red blood cells and hemoglobin of the mice, while the preferred traditional Chinese medicine composition tablet showed a significant effect on increasing red blood cells and hemoglobin. The results are shown in Tables 2 and 3.

[0091] Table 2. The results of red blood cell count of each Chinese medicine preparation on the 21st day in the thrombocytopenia model mice induced by guinea pig anti-platelet serum

[0092] "-" in the table represents that the red blood cell and hemoglobin levels of the corresponding group compared with the model control group are reduced or increased by less than 10%; "+" represents that the red blood cell and hemoglobin levels of the treatment group compared with the model control group are increased by 10-19%; "++" represents that the red blood cell and hemoglobin levels of the treatment group compared with the model control group are increased by 20-39%; "+++" represents that the red blood cell and hemoglobin levels of the treatment group compared with the model control group are increased by 40-59%; "++++" represents that the red blood cell and hemoglobin levels of the treatment group compared with the model control group are increased by more than 60%.

[0093] The red blood cell and hemoglobin levels of the treatment group compared with the model control group have statistical differences by Student's-t test: *, p<0.05; **, p<0.01; ***, p<0.001. The red blood cell and hemoglobin levels of the treatment group compared with the positive drug control group have statistical differences by Student's-t test: #, p<0.05; ##, p<0.01; ###, p<0.001.

[0094] Table 3. The results of hemoglobin levels of each Chinese medicine preparation on the 21st day in the thrombocytopenia model mice induced by guinea pig anti-platelet serum

[0095] "-" in the table represents that the red blood cell and hemoglobin levels of the corresponding group compared with the model control group are reduced or increased by less than 10%; "+" represents that the red blood cell and hemoglobin levels of the treatment group compared with the model control group are increased by 10-19%; "++" represents that the red blood cell and hemoglobin levels of the treatment group compared with the model control group are increased by 20-39%; "+++" represents that the red blood cell and hemoglobin levels of the treatment group compared with the model control group are increased by 40-59%; "++++" represents that the red blood cell and hemoglobin levels of the treatment group compared with the model control group are increased by more than 60%.

[0096] The red blood cell and hemoglobin levels of the treatment group compared with the model control group have statistical differences by Student's-t test: *, p<0.05; **, p<0.01; ***, p<0.001. The red blood cell and hemoglobin levels of the treatment group compared with the positive drug control group have statistical differences by Student's-t test: #, p<0.05; ##, p<0.01; ###, p<0.001.

[0097] Experimental conclusion: compared with the positive control drug eltrombopag, the prepared traditional Chinese medicine composition tablet has a significant effect of improving the peripheral red blood cell and hemoglobin levels of the immunogenic thrombocytopenia model mice. The test results show that when HPD300 macroporous adsorption resin is selected for purification, the 30% ethanol elution fraction of rhododendron is collected, and the mass ratio of rhododendron to scutellaria baicalensis is 7:3, the prepared tablet has the optimal effect of improving the peripheral platelet count of the immunogenic thrombocytopenia model mice,

[0098] The above is only a preferred specific embodiment of the present application, but the protection scope of the present application is not limited to this. Any person skilled in the art can easily think of changes or replacements within the technical range disclosed by the present application, which should be covered within the protection scope of the present application. Therefore, the protection scope of the present application should be subject to the protection scope of the claims.

Claims

1. A traditional Chinese medicine composition, characterized in that, is made from the following weight parts of traditional Chinese medicine raw materials: 1-10 parts by weight of Rhododendron simsii or extract thereof, 1-10 parts by weight of Scutellaria baicalensis or extract thereof.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, is made from the following weight parts of traditional Chinese medicine raw materials: 1-9 parts by weight of Rhododendron simsii or extract thereof, 1-10 parts by weight of Scutellaria baicalensis or extract thereof. Or, 1-7 parts by weight of Rhododendron simsii or extract thereof, 3-10 parts by weight of Scutellaria baicalensis or extract thereof. Or, 5-9 parts by weight of Rhododendron simsii or extract thereof, 5-10 parts by weight of Scutellaria baicalensis or extract thereof. Or, 7-9 parts by weight of Rhododendron simsii or extract thereof, 3-5 parts by weight of Scutellaria baicalensis or extract thereof. Or, 1-9 parts by weight of Rhododendron simsii or extract thereof, 1-9 parts by weight of Scutellaria baicalensis or extract thereof.

3. The traditional Chinese medicine composition according to any one of claims 1-2, characterized in that: is made from the following weight parts of traditional Chinese medicine raw materials: 9 parts by weight of Rhododendron simsii or extract thereof, 1 part by weight of Scutellaria baicalensis or extract thereof. Or, 7 parts by weight of Rhododendron simsii or extract thereof, 3 parts by weight of Scutellaria baicalensis or extract thereof. Or, 5 parts by weight of Rhododendron simsii or extract thereof, 5 parts by weight of Scutellaria baicalensis or extract thereof. Or, 3 parts by weight of Rhododendron simsii or extract thereof, 7 parts by weight of Scutellaria baicalensis or extract thereof. Or, 1 part by weight of Rhododendron simsii or extract thereof, 9 parts by weight of Scutellaria baicalensis or extract thereof.

4. The traditional Chinese medicine composition according to any one of claims 1-2, characterized in that: The Rhododendron simsii extract is prepared by the following method: Rhododendron simsii is crushed, sieved, an alcohol organic solvent is added as an extractant, reflux extraction is performed, the extract is combined, and the extract is concentrated under reduced pressure to obtain an extract, the extract is adsorbed by a macroporous resin column, deionized water is used for elution, and then different concentrations of alcohol organic solvents are used for elution, different concentrations of eluate are collected, and the Rhododendron simsii extract is obtained by concentration under reduced pressure. The Scutellaria baicalensis extract is prepared by the following method: Scutellaria baicalensis is crushed, sieved, an alcohol organic solvent is added as an extractant, reflux extraction is performed, the extract is combined, and the extract is concentrated under reduced pressure to obtain an extract, the extract is adsorbed by a macroporous resin column, deionized water is used for elution, and then different concentrations of alcohol organic solvents are used for elution, different concentrations of eluate are collected, and the Rhododendron simsii extract is obtained by concentration under reduced pressure. The macroporous resin column is selected from one of the following types: HPD-300, HPD-400, HPD-450, HPD-600, HPD-826, ADS-17, ADS-21, AB-8, D101, or D301.

5. The traditional Chinese medicine composition according to any one of claims 1-4, characterized in that: In the preparation of the Rhododendron simsii extract or the Scutellaria baicalensis extract, the ratio of Rhododendron simsii or Scutellaria baicalensis to the extractant is 1:1-12 kg / L, reflux extraction is performed 1-4 times, and the extraction time is 0.2-4 h each time, the alcohol organic solvent is methanol, ethanol, propanol, or butanol, and the mass fraction of the alcohol organic solvent is 5%-100%, 10%-80%, 20%-60%, or 40%-60%. Preferably, Rhododendron simsii or Scutellaria baicalensis is sieved to 18 mesh, the ratio of Rhododendron simsii or Scutellaria baicalensis to the extractant is 1:2-10 kg / L, reflux extraction is performed 1-3 times, and the extraction time is 0.5-2 h each time, the extract is concentrated under reduced pressure to a relative density of 1.10-1.30 (60°C), the alcohol organic solvent is ethanol, the mass fraction of the alcohol organic solvent is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 100%, and the amount of the alcohol organic solvent is 2-8 BV.

6. The traditional Chinese medicine composition according to any one of claims 1-4, characterized in that: In the preparation of the extract of Loropetalum chinense or the extract of Scutellaria baicalensis, the macroporous resin column is selected from one of the following types: HPD-300, HPD-400, ADS-17 or D101, and the adsorption flow rate is 1-8 mL·min -1 The macroporous resin has a diameter of 1-6 cm, a diameter-height ratio of 1:2-8, and a loading amount of 10-60 mL, and the static adsorption time is 2-24 h; the ionized water elution amount is 1BV-6BV, Preferably, the adsorption flow rate is 2-6 mL·min -1 Preferably, the macroporous resin has a diameter of 2-4 cm, a diameter-height ratio of 1:2-4, and a loading amount of 20-40 mL, and is statically adsorbed for 4-24 h; the ionized water elution amount is 2-6 BV.

7. The method for preparing the Chinese medicine composition according to any one of claims 1 to 6, characterized in that The preparation method is as follows: (1) Preparation of the extract of Loropetalum chinense: Loropetalum chinense is crushed, sieved, and added with an alcohol organic solvent as an extractant for reflux extraction. The extractants are combined and concentrated under reduced pressure to obtain an extract. The extract is adsorbed by a macroporous resin column, eluted with deionized water, discarded, and then eluted with alcohol organic solvents of different concentrations. The eluate of different concentrations is collected and concentrated under reduced pressure to obtain the extract of Loropetalum chinense; (2) Preparation of the extract of Scutellaria baicalensis: Scutellaria baicalensis is crushed, sieved, and added with an alcohol organic solvent as an extractant for reflux extraction. The extractants are combined and concentrated under reduced pressure to obtain an extract. The extract is adsorbed by a macroporous resin column, eluted with deionized water, discarded, and then eluted with alcohol organic solvents of different concentrations. The eluate of different concentrations is collected and concentrated under reduced pressure to obtain the extract of Loropetalum chinense; The macroporous resin column is selected from one of the following types: HPD-300, HPD-400, HPD-450, HPD-600, HPD-826, ADS-17, ADS-21, AB-8, D101, or D301; (3) The extract of Loropetalum chinense and the extract of Scutellaria baicalensis are mixed in a ratio of 1-10:1-10 by weight.

8. The preparation method according to claim 7, characterized in that: In the preparation of the extract of Loropetalum chinense in step (1) or the extract of Scutellaria baicalensis in step (2), the ratio of Loropetalum chinense or Scutellaria baicalensis to the extractant is 1:1-12 kg / L, and reflux extraction is performed 1-4 times, with each extraction time being 0.2-4 h. The alcohol organic solvent is methanol, ethanol, propanol, or butanol. The mass fraction of the alcohol organic solvent is 5%-100%, 10%-80%, 20%-60%, or 40%-60%. Preferably, Loropetalum chinense or Scutellaria baicalensis is sieved to 18 mesh. The ratio of Loropetalum chinense or Scutellaria baicalensis to the extractant is 1:2-10 kg / L, and reflux extraction is performed 1-3 times, with each extraction time being 0.5-2 h. The extract is concentrated under reduced pressure to a relative density of 1.10-1.30 (60°C). The alcohol organic solvent is ethanol. The mass fraction of the alcohol organic solvent is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 100%, and the amount is 2-8 BV. More preferably, the macroporous resin column is selected from one of the following models: HPD-300, HPD-400, ADS-17 or D101, and the adsorption flow rate is 1-8 mL·min -1 The macroporous resin has a diameter of 1-6 cm and a diameter-height ratio of 1:2-8, and the loading amount is 10-60 mL, and the static adsorption time is 2-24 h; the ion water elution amount is 1BV-6BV, Further preferably, the adsorption flow rate is 2-6 mL·min -1 The macroporous resin has a diameter of 2-4 cm, a diameter-height ratio of 1:2-4, a loading amount of 20-40 mL, and a static adsorption time of 4-24 h. The ionized water used for elution has a volume of 2-6 BV. In step (3), the ratio of the extract of Loropetalum chinense to the extract of Scutellaria baicalensis by weight is 9:1, 7:3, 5:5, 3:7, or 1:

9.

9. The traditional Chinese medicine composition according to any one of claims 1-6, characterized in that, The composition is in the form of a pharmaceutical preparation, and the pharmaceutical preparation can further include a pharmaceutical excipient as needed.

10. The traditional Chinese medicine composition of claim 9, wherein the pharmaceutical preparation is selected from tablets, capsules, buccal preparations, granules, pills, powders, pastes, Dan preparations, suspensions, solutions, injections, suppositories, pastes, sprays, drops, drop pills, or patches.

11. Use of the traditional Chinese medicine composition of any one of claims 1-6 in the preparation of a medicine for blood system diseases.

12. The use of claim 11, wherein the blood system disease includes, but is not limited to, iron deficiency anemia, aplastic anemia, acute leukemia, idiopathic thrombocytopenic purpura (ITP), primary immune thrombocytopenia, chronic anemia, or thrombocytopenia, preferably thrombocytopenia.

13. Use according to claim 11, characterized in that The medicine can promote peripheral platelet production and increase peripheral red blood cell and hemoglobin levels.

14. The traditional Chinese medicine composition of any one of claims 1-6, wherein The Loropetalum is Loropetalum chinense or Loropetalum rubrum. The Loropetalum is Loropetalum chinense or Loropetalum rub