Medical use of loropetalum chinense or extract thereof

Abstract

The use of Loropetalum chinense or an extract thereof in the preparation of a drug for circulatory system diseases. The Loropetalum chinense extract comprises a crude extract or a purified extract of Loropetalum chinense. The purified extract of Loropetalum chinense is obtained by means of: using medicinal materials of Loropetalum chinense (R. Br.) Oliver or Loropetalum chinense var. rubrum as raw materials, crushing same, adding water or an organic solvent as an extraction solvent to extract, performing concentration under reduced pressure and drying to obtain the crude extract of Loropetalum chinense; and adsorbing the crude extract on a macroporous resin, and eluting same with ethanol at different concentrations. The extract is used for treatment of thrombocytopenia, and can promote peripheral platelet production and / or enhance platelet activity.

Description

Medical use of Loropetalum chinense or extract thereof TECHNICAL FIELD

[0001] The present application belongs to the field of traditional Chinese medicine extract, and particularly relates to a medical use of Loropetalum chinense or extract 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 chinense is usually a shrub or a small tree, and is widely distributed in central, southern and southwestern provinces, and also has rich wild resources in China. The flower, leaf and root of Loropetalum chinense can be used as medicine, and has the effects of clearing heat and detoxifying, astringing, hemostasis, etc. Loropetalum chinense is commonly used for the treatment of burns, scalds, external injuries, hemorrhage, hemoptysis, metrorrhagia, diarrhea and gastric ulcer. At present, more than 60 compounds have been isolated from Loropetalum chinense, including tannins, flavonoid glycosides, lignans and steroids, which have the effects of antibacterial, anti-inflammatory, antioxidant, promoting healing, etc. Loropetalum chinense has high clinical application value and broad market prospect.

[0003] Thrombocytopenia is a refractory blood system disease, mainly including primary immune thrombocytopenia, secondary thrombocytopenia caused by diseases such as malignant tumor, systemic lupus erythematosus and chronic anemia, and drug-induced thrombocytopenia caused by chemotherapy drugs, thrombolytic drugs, etc. The incidence of thrombocytopenia in China is relatively high (5-10 per 100,000 people), and patients generally have symptoms such as skin purpura, fatigue, anxiety, and may also have leukopenia, anemia and total hemoglobin reduction, etc. Severe cases can die due to internal bleeding or intracranial hemorrhage. The etiology of thrombocytopenia is complex, and the pathogenesis has not been completely analyzed. 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 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.

[0004] The researches on L. alba and L. indica in China mainly focus on the separation of compounds and medicinal activities. Zhang Wugang et al. once recorded that L. alba leaves have the effects of "relaxing muscles, promoting blood circulation, relieving fever and diarrhea, treating stomachache, muscle spasm, diarrhea, callus and external injury bleeding" (Analysis of Hemostatic Chemical Constituents of L. alba, Chinese Journal of Experimental Traditional Chinese Formulas, 2017, 23(05): 47-52) and other effects. Lian Zeqin et al. reported that L. alba can promote the repair of damaged epithelial tissue and converge wound, and has significant and accurate effects of promoting hemostasis and coagulation (Preliminary Study on the Substances of L. alba for Promoting Wound Healing in Rats, Chinese Journal of Chinese Materia Medica, 2013, 38(20): 3566-3570). L. alba or its extract is also widely reported to be clinically used for treating external injury bleeding, postpartum poor uterine contraction and bleeding, and promoting tissue healing at a surgical site. At present, there is no relevant research on the prevention and treatment of thrombocytopenia by L. alba or its extract. The inventors found that L. alba or its extract can be used as a drug for thrombocytopenia and can promote peripheral platelet production or improve platelet activity. SUMMARY

[0005] The purpose of the present application is to provide a medical use of L. alba or its extract.

[0006] The first purpose of the present application is to provide an application of traditional Chinese medicine L. alba or L. alba extract in preparing a drug for blood system diseases.

[0007] The blood system diseases include but are not limited to thrombocytopenia, iron deficiency anemia, aplastic anemia, acute leukemia, idiopathic thrombocytopenic purpura (ITP), primary immune thrombocytopenia or chronic anemia, and preferably thrombocytopenia.

[0008] The drug can promote peripheral platelet production or improve platelet activity.

[0009] The traditional Chinese medicine L. alba is selected from L. alba and / or L. indica, and the L. alba extract is selected from L. alba extract and / or L. indica extract.

[0010] The L. alba extract includes L. alba crude extract and / or L. alba purified extract.

[0011] The L. alba crude extract is obtained by extracting the traditional Chinese medicine L. alba with water or an alcohol aqueous solution.

[0012] The L. alba crude extract is prepared by the following method: step (1), crushing L. alba and adding water or an organic solvent as an extractant; step (2), reflux extraction and vacuum concentration to obtain the L. alba crude extract.

[0013] The crude extract of the Loropetalum chinense is prepared by the following method: (1) crushing and sieving the Loropetalum chinense, adding water or an alcohol organic solvent or an ester organic solvent or a ketone organic solvent as an extractant, and standing; (2) reflux extraction, combining the extract, and reducing pressure concentration to obtain the crude extract of the Loropetalum chinense.

[0014] In the step (1), the ratio of the Loropetalum chinense to the extractant is 1:1-12 kg / L, and the standing time is 0.5-18 h; in the step (2), the reflux extraction is performed 1-4 times, and each extraction time is 0.2-4 h.

[0015] Preferably, in the step (1), the sieving is 18 mesh, the ratio of the Loropetalum chinense to the extractant is 1:2-10 kg / L, and the standing time is 1-12 h; in the step (2), the reflux extraction is performed 1-3 times, and each extraction time is 0.5-2 h, and the extract is reduced pressure concentrated to a relative density of 1.10-1.30 (60℃).

[0016] In the step (1), the alcohol organic solvent is methanol, ethanol, propanol or butanol, the ester organic solvent is ethyl formate, ethyl acetate, propyl acetate, isopropyl acetate or butyl acetate, and the ketone organic solvent is acetone, butanone or pentanone.

[0017] Preferably, the mass fraction of the alcohol organic solvent is 5%-100%, 10%-80%, 20%-60% or 40%-60%.

[0018] Further preferably, the mass fraction of the alcohol organic solvent is 20%, 40%, 60%, 80% or 100%.

[0019] In the crude extract of the Loropetalum chinense, the total phenol content is in the range of 1%-99%, 5%-95%, 6%-90%, 7%-85%, 8%-80%, 9%-78%, 10%-76%, 11%-74%, 12%-70%, 13%-68%, 14%-66%, 15%-64%, 16%-62%, 18%-60%, 20%-58%, 22%-56%, 24%-54%, 26%-52%, 28%-50%, 30%-48%, 32%-46%, 34%-44%, 32%-42%, 34%-40% or 36%-38%.

[0020] Preferably, the total phenol content of the crude extract of the Magnolia officinalis is 7.3%, 19.4%, 25.4%, 31.3%, 20.1%, 23.1%, 6.6%, 5.8%, 7.3%, 15.2%, 22.3%, 30.2%, 19.4%, 21.4%, 7.0%, 5.3%, 17.8%, 62.3%, 50.4%, 42.1%, 15.4%, 70.2%, 58.7%, 43.3%, 10.2%, 55.7%, 48.9%, 40.3%, 19.1%, 58.2%, 55.9%, or 38.9%.

[0021] A second object of the present application is that the purified extract of Magnolia officinalis is prepared by the following method: the crude extract of Magnolia officinalis is purified by a macroporous resin column to obtain a purified extract of Magnolia officinalis with different polarities.

[0022] The purified extract of Magnolia officinalis is prepared by the following method: the crude extract of Magnolia officinalis is diluted with distilled water, adsorbed by a macroporous resin column, eluted with deionized water, discarded, and then eluted with different concentrations of alcohol organic solvents, collected, concentrated under reduced pressure, and dried to obtain the purified extract of Magnolia officinalis; the macroporous resin column is independently selected from a macroporous resin column with a type number of HPD-300, HPD-400, HPD-450, HPD-600, HPD-826, ADS-17, ADS-21, AB-8, D101, or D301.

[0023] The macroporous resin column is selected from one of the following type numbers: AB-8, HPD-300, ADS-17, or D101; 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 with a static adsorption time of 2-24 h; the deionized water elution amount is 1BV-6BV, the alcohol organic solvent is ethanol with a mass fraction of 10%-80%, and the amount is 2-10BV.

[0024] Preferably, the adsorption flow rate is 2-6 mL·min -1 ; the macroporous resin has a diameter of 2-4 cm and a diameter-height ratio of 1:2-4, the loading amount is 20-40 mL with a static adsorption time of 4-24 h; the deionized water elution amount is 2-6BV; the gradient elution mass fraction of ethanol is 10%, 30%, 50%, or 70%, and the amount is 2-8BV.

[0025] More preferably, the adsorption flow rate is 5 mL·min -1The macroporous resin has a diameter of 4 cm, a diameter-height ratio of 1:4 or 1:8, a loading amount of 20 mL or 40 mL, a static adsorption time of 12 h, the ionized water elution amount is 2 BV, and the ethanol gradient elution mass fraction is 10%, 30%, 50% or 70%, and the amount is 5 BV or 6 BV.

[0026] The total phenol content in the purified extract of Rhododendron simsii is 1%-99%, 5%-95%, 6%-90%, 7%-85%, 8%-80%, 9%-78%, 10%-76%, 11%-74%, 12%-70%, 13%-68%, 14%-66%, 15%-64%, 16%-62%, 18%-60%, 20%-58%, 22%-56%, 24%-54%, 26%-52%, 28%-50%, 30%-48%, 32%-46%, 34%-44%, 32%-42%, 34%-40% or 36%-38%.

[0027] Preferably, the total phenol content in the purified extract of Rhododendron simsii is 17.8%, 62.3%, 50.4%, 42.1%, 15.4%, 70.2%, 58.7%, 43.3%, 10.2%, 55.7%, 48.9%, 40.3%, 19.1%, 58.2% or 38.9%.

[0028] The medicine is a medicine composition containing traditional Chinese medicine Rhododendron simsii or Rhododendron simsii extract.

[0029] The medicine composition can further contain a pharmaceutically acceptable carrier.

[0030] The medicine composition is prepared into any pharmaceutically acceptable dosage form.

[0031] The pharmaceutically acceptable dosage form is selected from tablets, capsules, solutions, granules, powders, ointments, pills, suspensions, suppositories, rubs, emulsions, smears, patches or sprays.

[0032] [According to the rules 91 correction 23.07.2025] The dosage form is in unit dosage form, and each unit dosage contains 1-10 mg, or 10-100 mg, or 100-1000 mg, or 1000-10000 mg of traditional Chinese medicine Rhododendron simsii or Rhododendron simsii extract.

[0033] The following is the explanation and description of the terms of the present application:

[0034] The relevant information of Loropetalum chinense is as follows: Loropetalum chinense (R. Brown) Oliv. is a plant of Hamamelidaceae Hamamelis genus. It has the effects of astringing, hemostasis, clearing heat and detoxification, and stopping diarrhea. It is used to treat various bleeding syndromes such as epistaxis, hemoptysis, trauma, and so on, as well as scalding, diarrhea, and dysentery.

[0035] The term "pharmaceutically acceptable" means that the carrier, vehicle, diluent, adjuvant, and / or formed salt is generally chemically and / or physically compatible with the other ingredients constituting a pharmaceutical formulation and physiologically compatible with the recipient thereof.

[0036] [Corrected according to Rule 91 on 23.07.2025] The pharmaceutical composition of the present application can be in any form of reusable pharmaceutical preparation, such as oral, injection, external use, etc. Oral dosage forms include but are not limited to tablets, capsules, oral liquids, granules, pills, suspensions, injections selected from water injections, powder injections, and external preparations selected from patches and ointments. All preparations can be prepared according to conventional techniques of pharmacy, such as using any one of the compounds of the present application, or its stereoisomer, or its pharmaceutically acceptable salt as the active pharmaceutical ingredient, if necessary, adding a pharmaceutically acceptable carrier to prepare the above-mentioned pharmaceutical dosage forms suitable for administration, wherein the unit dose of the active pharmaceutical ingredient can be 0.1 mg-10000 mg, such as 0.1 mg-10000 mg of active pharmaceutical ingredient per tablet, preferably 5-500 mg of active pharmaceutical ingredient, or 1-10 mg of active pharmaceutical ingredient, or 10-100 mg of active pharmaceutical ingredient, or 100-1000 mg of active pharmaceutical ingredient, or 1000-10000 mg of active pharmaceutical ingredient.

[0037] Obviously, according to the above content of the present application, other various forms of modifications, substitutions or changes can be made according to the ordinary technical knowledge and common practice in the art without departing from the above-mentioned basic technical ideas of the present application.

[0038] The beneficial technical effects of the present application are: 1. In the prior art, it is believed that the promotion of platelet production mainly depends on the stimulation of bone marrow megakaryocyte differentiation and maturation, the promotion of platelet production, and the reduction of megakaryocyte and platelet destruction. The present application finds that the Loropetalum chinense extract obtained by a certain extraction and purification process can significantly increase the peripheral blood platelet level of a platelet reduction model animal and promote peripheral blood platelet production. Unlike the body's main hemostasis and coagulation mechanism through a series of mechanisms such as contraction of blood vessel wall, stimulation of platelet aggregation, activation of coagulation factors and fibrinolysis system.

[0039] 2. The application provides the application of E. acuminatum and E. indica extracts in promoting peripheral platelet production. Using a chemotherapy drug-induced thrombocytopenia mouse model, the E. acuminatum and E. indica extract prepared by the extraction method of the application can significantly promote the peripheral platelet production of the model mice, and the application of the E. acuminatum and E. indica extracts has not been reported in the prior art. DETAILED DESCRIPTION

[0040] The application is further illustrated by the following examples, but is not limited to the application.

[0041] Example 1: Preparation of E. acuminatum crude extract and determination of total phenol content

[0042] (1) 500 g of fresh E. acuminatum material was crushed and then passed through an 18-mesh sieve;

[0043] Then water was added as an extraction solvent, and it was recorded as E. acuminatum crude extract No. 1 (referred to as white crude 1);

[0044] 20% ethanol was added as an extraction solvent, and it was recorded as E. acuminatum crude extract No. 2 (referred to as white crude 2);

[0045] 40% ethanol was added as an extraction solvent, and it was recorded as E. acuminatum crude extract No. 3 (referred to as white crude 3);

[0046] 60% ethanol was added as an extraction solvent, and it was recorded as E. acuminatum crude extract No. 4 (referred to as white crude 4);

[0047] 80% ethanol was added as an extraction solvent, and it was recorded as E. acuminatum crude extract No. 5 (referred to as white crude 5);

[0048] 100% methanol was added as an extraction solvent, and it was recorded as E. acuminatum crude extract No. 6 (referred to as white crude 6);

[0049] Acetone was added as an extraction solvent, and it was recorded as E. acuminatum crude extract No. 7 (referred to as white crude 7);

[0050] Ethyl acetate was added as an extraction solvent, and it was recorded as E. acuminatum crude extract No. 8 (referred to as white crude 8);

[0051] The ratio of each group of material liquid was 1:2 kg / L, and it was allowed to stand for 12 h, and then it was continuously extracted by condensation reflux for 3 times, 0.5 h each time, the extraction liquid was combined, and the extraction liquid was concentrated under reduced pressure to a relative density of 1.10-1.30 (60℃), and the following E. acuminatum crude extracts were obtained: E. acuminatum crude extract No. 1, E. acuminatum crude extract No. 2, E. acuminatum crude extract No. 3, E. acuminatum crude extract No. 4, E. acuminatum crude extract No. 5, E. acuminatum crude extract No. 6, E. acuminatum crude extract No. 7 and E. acuminatum crude extract No. 8.

[0052] (2) 500 g of fresh materials of E. alba were ground and then passed through an 18-mesh sieve, respectively;

[0053] Then, water was added to the powder as an extraction solvent, and the crude extract of E. alba No. 9 (referred to as white crude 9) was recorded;

[0054] 20% ethanol was added as an extraction solvent, and the crude extract of E. alba No. 10 (referred to as white crude 10) was recorded;

[0055] 40% ethanol was added as an extraction solvent, and the crude extract of E. alba No. 11 (referred to as white crude 11) was recorded;

[0056] 60% ethanol was added as an extraction solvent, and the crude extract of E. alba No. 12 (referred to as white crude 12) was recorded;

[0057] 80% ethanol was added as an extraction solvent, and the crude extract of E. alba No. 13 (referred to as white crude 13) was recorded;

[0058] 100% methanol was added as an extraction solvent, and the crude extract of E. alba No. 14 (referred to as white crude 14) was recorded;

[0059] Acetone was added as an extraction solvent, and the crude extract of E. alba No. 15 (referred to as white crude 15) was recorded;

[0060] Ethyl acetate was added as an extraction solvent, and the crude extract of E. alba No. 16 (referred to as white crude 16) was recorded;

[0061] The ratio of each group of materials to liquid was 1:10 kg / L, and the mixture was allowed to stand for 1 h, then continuously condensed and refluxed for 1 time, each time for 2 h, the extract was combined, and the extract was concentrated under reduced pressure to a relative density of 1.10-1.30 (60°C); the following crude extracts of E. alba were obtained: crude extract of E. alba No. 9, crude extract of E. alba No. 10, crude extract of E. alba No. 11, crude extract of E. alba No. 12, crude extract of E. alba No. 13, crude extract of E. alba No. 14, crude extract of E. alba No. 15, and crude extract of E. alba No. 16.

[0062] (3) The total phenol content of each crude extract of E. alba was detected by Folin phenol colorimetry with gallic acid as the standard. The yield and total phenol content of the crude extract of E. alba were calculated according to the following formula, and the total phenol content of the crude extract of E. alba was calculated as gallic acid, as shown in Table 1.

[0063] Yield of crude extract of E. alba % = dry extract of E. alba / E. alba feed amount * 100%

[0064] Total phenol content of crude extract of E. alba % = total phenol mass of crude extract of E. alba / dry extract of crude extract of E. alba * 100%

[0065] Table 1 Yield and total phenol content of crude extract of E. alba prepared by different extraction processes

[0066] Conclusion: The yield of the crude extract of Acacia confusa is between 11.5% and 65.8%, and the total phenol content is between 5.7% and 30.8%. The total phenol content of the crude extract of Acacia confusa No. 4 is the highest.

[0067] Example 2 Preparation of crude extract of Acacia confusa and determination of total phenol content

[0068] (1) 500g of fresh Acacia confusa material was ground and then passed through an 18-mesh sieve;

[0069] Then water was added to the powder as an extraction solvent, and it was recorded as crude extract of Acacia confusa No. 1 (abbreviated as red crude 1);

[0070] 20% ethanol was added as an extraction solvent, and it was recorded as crude extract of Acacia confusa No. 2 (abbreviated as red crude 2);

[0071] 40% ethanol was added as an extraction solvent, and it was recorded as crude extract of Acacia confusa No. 3 (abbreviated as red crude 3);

[0072] 60% ethanol was added as an extraction solvent, and it was recorded as crude extract of Acacia confusa No. 4 (abbreviated as red crude 4);

[0073] 80% ethanol was added as an extraction solvent, and it was recorded as crude extract of Acacia confusa No. 5 (abbreviated as red crude 5);

[0074] 100% methanol was added as an extraction solvent, and it was recorded as crude extract of Acacia confusa No. 6 (abbreviated as red crude 6);

[0075] Acetone was added as an extraction solvent, and it was recorded as crude extract of Acacia confusa No. 7 (abbreviated as red crude 7);

[0076] Ethyl acetate was added as an extraction solvent, and it was recorded as crude extract of Acacia confusa No. 8 (abbreviated as red crude 8).

[0077] The ratio of each group of material to liquid was 1:2 kg / L, and it was allowed to stand for 12h, then it was continuously condensed and refluxed for 3 times, 1h each time, and the extraction liquid was combined and concentrated under reduced pressure to a relative density of 1.10-1.30 (60℃), and the following crude extracts of Acacia confusa were obtained: crude extract of Acacia confusa No. 1, crude extract of Acacia confusa No. 2, crude extract of Acacia confusa No. 3, crude extract of Acacia confusa No. 4, crude extract of Acacia confusa No. 5, crude extract of Acacia confusa No. 6, crude extract of Acacia confusa No. 7, and crude extract of Acacia confusa No. 8.

[0078] (2) 500g of fresh Acacia confusa material was ground and then passed through an 18-mesh sieve;

[0079] Then water was added to the powder as an extraction solvent, and it was recorded as crude extract of Acacia confusa No. 9 (abbreviated as red crude 9);

[0080] The 20% ethanol was added as the extraction solvent, and the obtained product was recorded as No. 10 of the crude extract of E. acuminatum (abbreviated as red crude 10);

[0081] The 40% ethanol was added as the extraction solvent, and the obtained product was recorded as No. 11 of the crude extract of E. acuminatum (abbreviated as red crude 11);

[0082] The 60% ethanol was added as the extraction solvent, and the obtained product was recorded as No. 12 of the crude extract of E. acuminatum (abbreviated as red crude 12);

[0083] The 80% ethanol was added as the extraction solvent, and the obtained product was recorded as No. 13 of the crude extract of E. acuminatum (abbreviated as red crude 13);

[0084] The 100% methanol was added as the extraction solvent, and the obtained product was recorded as No. 14 of the crude extract of E. acuminatum (abbreviated as red crude 14);

[0085] The acetone was added as the extraction solvent, and the obtained product was recorded as No. 15 of the crude extract of E. acuminatum (abbreviated as red crude 15);

[0086] The ethyl acetate was added as the extraction solvent, and the obtained product was recorded as No. 16 of the crude extract of E. acuminatum (abbreviated as red crude 16).

[0087] The liquid-to-solid ratio of each group was 1:12 kg / L, and the mixture was allowed to stand for 1 h, and then was subjected to continuous condensation reflux extraction for 1 time, each time for 2 h. The extraction liquid was combined and concentrated under reduced pressure to a relative density of 1.10-1.30 (60°C), to obtain the following crude extracts of E. acuminatum: No. 9 of the crude extract of E. acuminatum, No. 10 of the crude extract of E. acuminatum, No. 11 of the crude extract of E. acuminatum, No. 12 of the crude extract of E. acuminatum, No. 13 of the crude extract of E. acuminatum, No. 14 of the crude extract of E. acuminatum, No. 15 of the crude extract of E. acuminatum, and No. 16 of the crude extract of E. acuminatum.

[0088] (3) The total phenol content of each crude extract of E. acuminatum was detected by the Folin phenol colorimetric method using gallic acid as the standard. The yield and the total phenol content of the crude extract of E. acuminatum were calculated according to the following formula. The total phenol content of the crude extract of E. acuminatum was shown in Table 2.

[0089] The yield of the crude extract of E. acuminatum (%) = the dry extract amount of the crude extract of E. acuminatum / the E. acuminatum feeding amount * 100%

[0090] The total phenol content of the crude extract of E. acuminatum (%) = the total phenol mass of the crude extract of E. acuminatum / the dry extract amount of the crude extract of E. acuminatum * 100%

[0091] Table 2 The yield and the total phenol content of the crude extract of E. acuminatum prepared by different extraction processes

[0092] Conclusion: The yield of the crude extract of E. acuminatum was between 10.3% and 72.9%, and the total phenol content was between 5.3% and 31.3%. The total phenol content of the crude extract of E. acuminatum No. 4 was the highest.

[0093] Example 3, Preparation of White Mixture of Purified Extract Components and Determination of Total Phenol Content

[0094] (1) The extract white crude 4 in Example 1 was redissolved with distilled water at a flow rate of 5 mL·min-1 through a macroporous resin column for adsorption. The type of macroporous adsorption resin used was AB-8, the diameter was 4 cm, the diameter-height ratio was 1:4, the total sample amount was 20 mL, static adsorption was 12 h, and first deionized water was used for elution. The amount of distilled water used was 2 BV, and the distilled water part was discarded. -1

[0095] 10% ethanol was used for elution, the amount of 10% ethanol used was 5 BV, and the component was collected and recorded as White Mixture of Purified Extract Component No. 1 (abbreviated as white pure 1).

[0096] 30% ethanol was used for elution, the amount of 30% ethanol used was 6 BV, and the component was collected and recorded as White Mixture of Purified Extract Component No. 2 (abbreviated as white pure 2).

[0097] 50% ethanol was used for elution, the amount of 50% ethanol used was 6 BV, and the component was collected and recorded as White Mixture of Purified Extract Component No. 3 (abbreviated as white pure 3).

[0098] 70% ethanol was used for elution, the amount of 70% ethanol used was 5 BV, and the component was collected and recorded as White Mixture of Purified Extract Component No. 4 (abbreviated as white pure 4).

[0099] (2) The extract white crude 4 in Example 1 was redissolved with distilled water at a flow rate of 5 mL·min-1 through a macroporous resin column for adsorption. The type of macroporous adsorption resin used was HPD-300, the diameter was 4 cm, the diameter-height ratio was 1:8, the total sample amount was 40 mL, static adsorption was 12 h, and first deionized water was used for elution. The amount of distilled water used was 2 BV, and the distilled water part was discarded.

[0100] Then 10% ethanol was used for elution, the amount of 10% ethanol used was 5 BV, and the component was collected and recorded as White Mixture of Purified Extract Component No. 5 (abbreviated as white pure 5).

[0101] 30% ethanol was used for elution, the amount of 30% ethanol used was 6 BV, and the component was collected and recorded as White Mixture of Purified Extract Component No. 6 (abbreviated as white pure 6).

[0102] 50% ethanol was used for elution, the amount of 50% ethanol used was 6 BV, and the component was collected and recorded as White Mixture of Purified Extract Component No. 7 (abbreviated as white pure 7).

[0103] 70% ethanol was used for elution, the amount of 70% ethanol used was 5 BV, and the component was collected and recorded as White Mixture of Purified Extract Component No. 8 (abbreviated as white pure 8).

[0104] ​(3) The extract white crude 4 in Example 1 was redissolved with distilled water, and was adsorbed by a macroporous resin column at a flow rate of 5 mL·min-1. The type of macroporous adsorption resin used was ADS-17, the diameter was 4 cm, the ratio of diameter to height was 1:4, the total amount of extract loaded was 20 mL, and static adsorption was performed for 12 h. Deionized water was used for elution, the amount of distilled water used was 2 BV, the distilled water part was discarded, -1

[0105] Then 10% ethanol was used for elution, the amount of 10% ethanol used was 5 BV, and the component collected was recorded as white flower kanchuang purified component 9 (abbreviated as white pure 9) ;

[0106] 30% ethanol was used for elution, the amount of 30% ethanol used was 6 BV, and the component collected was recorded as white flower kanchuang purified component 10 (abbreviated as white pure 10) ;

[0107] 50% ethanol was used for elution, the amount of 50% ethanol used was 6 BV, and the component collected was recorded as white flower kanchuang purified component 11 (abbreviated as white pure 11) ;

[0108] 70% ethanol was used for elution, the amount of 70% ethanol used was 5 BV, and the component collected was recorded as white flower kanchuang purified component 12 (abbreviated as white pure 12).

[0109] (4) The extract white crude 4 in Example 1 was redissolved with distilled water, and was adsorbed by a macroporous resin column at a flow rate of 5 mL·min-1. The type of macroporous adsorption resin used was D101, the diameter was 4 cm, the ratio of diameter to height was 1:8, the total amount of extract loaded was 40 mL, and static adsorption was performed for 12 h. Deionized water was used for elution, the amount of distilled water used was 2 BV, the distilled water part was discarded,

[0110] 10% ethanol was used for elution, the amount of 10% ethanol used was 5 BV, and the component collected was recorded as white flower kanchuang purified 13 (abbreviated as white pure 13) ;

[0111] 30% ethanol was used for elution, the amount of 30% ethanol used was 6 BV, and the component collected was recorded as white flower kanchuang purified 14 (abbreviated as white pure 14) ;

[0112] 50% ethanol was used for elution, the amount of 50% ethanol used was 6 BV, and the component collected was recorded as white flower kanchuang purified component 15 (abbreviated as white pure 15) ;

[0113] 70% ethanol was used for elution, the amount of 70% ethanol used was 5 BV, and the component collected was recorded as white flower kanchuang purified component 16 (abbreviated as white pure 16).

[0114] (5) Then, each of the white flower kanchuang purified components 1-16 was concentrated and dried, and white flower kanchuang purified components were successfully prepared.

[0115] ​(6) Folin-phenol colorimetric method was used to detect the total phenol content of each purified L. alba product with gallic acid as the standard. The transfer rate and total phenol content of the purified L. alba product were calculated according to the following formula, as shown in Table 3.

[0116] Transfer rate of purified L. alba product = total phenol mass of purified L. alba product / total phenol mass of L. alba crude extract * 100%

[0117] Total phenol content of purified L. alba product = total phenol mass of purified L. alba product / dry extract mass of purified L. alba product * 100%

[0118] Table 3. Transfer rate and total phenol content of purified L. alba product prepared by different purification processes

[0119] Conclusion: The transfer rate of the purified L. alba product prepared in this example ranged from 29.9% to 85.8%, and the total phenol content ranged from 10.2% to 70.2%.

[0120] Example 4, Preparation of L. formosana Purified Components and Determination of Total Phenol Content

[0121] (1) The L. formosana crude extract 4 in Example 2 was resuspended with distilled water and adsorbed on a macroporous resin column at a flow rate of 2 mL·min-1. The macroporous adsorption resin used was HPD-300, with a diameter of 4 cm and a diameter-height ratio of 1:4. The total sample amount was 20 mL, and the static adsorption time was 12 h. First, deionized water was used for elution, and the amount of distilled water used was 2 BV, which was discarded;

[0122] Then 10% ethanol was used for elution, and the amount of 10% ethanol used was 5 BV. The collected component was designated as L. formosana purified component No. 1 (abbreviated as red pure 1);

[0123] 30% ethanol was used for elution, and the amount of 30% ethanol used was 6 BV. The collected component was designated as L. formosana purified component No. 2 (abbreviated as red pure 2);

[0124] 50% ethanol was used for elution, and the amount of 50% ethanol used was 6 BV. The collected component was designated as L. formosana purified component No. 3 (abbreviated as red pure 3);

[0125] 70% ethanol was used for elution, and the amount of 70% ethanol used was 5 BV. The collected component was designated as L. formosana purified component No. 4 (abbreviated as red pure 4).

[0126] (2) The red coarse 4 extract in Example 2 was selected and redissolved with distilled water, and was adsorbed by a macroporous resin column at a flow rate of 2 mL·min-1. The selected macroporous adsorption resin was HPD-400, the diameter was 4 cm, the diameter-height ratio was 1:8, the total sample amount was 40 mL, and static adsorption was performed for 12 h. First, deionized water was used for elution, and the amount of distilled water was 2 BV, and the distilled water part was discarded.

[0127] Then 10% ethanol was used for elution, the amount of 10% ethanol was 5 BV, and the collected component was recorded as No. 5 of the purified component of Rhododendron simsii (abbreviated as red pure 5);

[0128] 30% ethanol was used for elution, the amount of 30% ethanol was 6 BV, and the collected component was recorded as No. 6 of the purified component of Rhododendron simsii (abbreviated as red pure 6);

[0129] 50% ethanol was used for elution, the amount of 50% ethanol was 6 BV, and the collected component was recorded as No. 7 of the purified component of Rhododendron simsii (abbreviated as red pure 7);

[0130] 70% ethanol was used for elution, the amount of 70% ethanol was 5 BV, and the collected component was recorded as No. 8 of the purified component of Rhododendron simsii (abbreviated as red pure 8).

[0131] (3) The red coarse 4 extract in Example 2 was selected and redissolved with distilled water, and was adsorbed by a macroporous resin column at a flow rate of 2 mL·min-1. The selected macroporous adsorption resin was D101, the diameter was 4 cm, the diameter-height ratio was 1:4, the total sample amount was 20 mL, and static adsorption was performed for 12 h. First, deionized water was used for elution, and the amount of distilled water was 2 BV, and the distilled water part was discarded.

[0132] Then 10% ethanol was used for elution, the amount of 10% ethanol was 5 BV, and the collected component was recorded as No. 9 of the purified component of Rhododendron simsii (abbreviated as red pure 9);

[0133] 30% ethanol was used for elution, the amount of 30% ethanol was 6 BV, and the collected component was recorded as No. 10 of the purified component of Rhododendron simsii (abbreviated as red pure 10);

[0134] 50% ethanol was used for elution, the amount of 50% ethanol was 6 BV, and the collected component was recorded as No. 11 of the purified component of Rhododendron simsii (abbreviated as red pure 11);

[0135] 70% ethanol was used for elution, the amount of 70% ethanol was 5 BV, and the collected component was recorded as No. 12 of the purified component of Rhododendron simsii (abbreviated as red pure 12).

[0136] (4) The red rough 4 extract in Example 2 was redissolved with distilled water, and was adsorbed by a macroporous resin column at a flow rate of 2 mL·min-1. The macroporous adsorption resin used was D301, the diameter was 4 cm, the diameter-height ratio was 1:8, and the total sample amount was 40 mL. The static adsorption time was 12 h. First, deionized water was used for elution, and the amount of distilled water was 2 BV. The distilled water part was discarded.

[0137] Then 10% ethanol was used for elution, and the amount of 10% ethanol was 5 BV. The collected component was recorded as No. 13 of the purified Rhododendron simsii (referred to as red pure 13 for short);

[0138] 30% ethanol was used for elution, and the amount of 30% ethanol was 6 BV. The collected component was recorded as No. 14 of the purified Rhododendron simsii (referred to as red pure 14, the same hereinafter);

[0139] 50% ethanol was used for elution, and the amount of 50% ethanol was 6 BV. The collected component was recorded as No. 15 of the purified Rhododendron simsii (referred to as red pure 15, the same hereinafter);

[0140] 70% ethanol was used for elution, and the amount of 70% ethanol was 5 BV. The collected component was recorded as No. 16 of the purified Rhododendron simsii (referred to as red pure 16, the same hereinafter).

[0141] (5) Then each of the above purified Rhododendron simsii components 1-16 was concentrated and dried, and the purified Rhododendron simsii components were successfully prepared.

[0142] (6) The total phenol content of each purified Rhododendron simsii was detected by Folin phenol colorimetry with gallic acid as a standard. The transfer rate and total phenol content of the purified Rhododendron simsii were calculated according to the following formula, as shown in Table 4.

[0143] The transfer rate of the purified Rhododendron simsii was calculated according to the following formula: the total phenol mass of the purified Rhododendron simsii / the total phenol mass of the crude Rhododendron simsii*100%

[0144] The total phenol content of the purified Rhododendron simsii was calculated according to the following formula: the total phenol mass of the purified Rhododendron simsii / the dry extract amount of the purified Rhododendron simsii*100%

[0145] Table 4. The transfer rate and total phenol content of the purified Rhododendron simsii prepared by different purification processes

[0146] Conclusion: The transfer rate of the purified Rhododendron simsii prepared in this example was in the range of 38.7%-84.2%, and the total phenol content was in the range of 10.1%-67.1%.

[0147] Example 5 Preparation of granules

[0148] Take any one of the extract components prepared in Examples 1-4 1100g, add the appropriate amount of lactose, mannitol and a little microcrystalline cellulose, mix well, spray into 90% ethanol solution, granulate, dry at 60°C, and prepare 1500g granules, which are granules.

[0149] Tablet preparation of Example 6

[0150] Take any one of the extract components prepared in Examples 1-4 100g, liquid paraffin 10g, talc 25g, starch 30g, tartaric acid 25g, and prepare 1000 tablets by tabletting.

[0151] Capsule preparation of Example 7

[0152] The capsules of this example are composed of a drug solution and a gel solution. Take any one of the extract components prepared in Examples 1-4 100g, add vitamin E 10g as an antioxidant and Tween 80 10g 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 to the gel tank in turn, heat to 80°C, then add gelatin and continuously stir and vacuum until the gelatin is completely dissolved. Filter the gel solution and store the filtrate at 60°C.

[0153] Pharmacodynamic Example 1 Promotion of platelet production in a model of thrombocytopenia induced by chemotherapy drugs by different purified components of A. luteoviridis and A. sanguinea

[0154] Objective: To observe the effect of orally administered different purified components of A. luteoviridis and A. sanguinea on the peripheral blood of a model of thrombocytopenia induced by chemotherapy drugs in mice, and to demonstrate their role in promoting platelet production.

[0155] Test substances: White crude 1, white crude 6, white crude 7, white crude 8, red crude 1, red crude 6, red crude 7, red crude 8 from Examples 1 and 2, and white pure 2, white pure 3, white pure 6, white pure 7, white pure 10, white pure 11, white pure 14, white pure 15, red pure 2, red pure 3, red pure 6, red pure 7, red pure 10, red pure 11, red pure 14, red pure 15 from Examples 3 and 4 were dissolved in distilled water to prepare oral preparations. In addition, we also prepared A. luteoviridis extract according to the preparation method in the literature (Zhang Wugang et al., Analysis of Hemostatic Chemical Components of A. luteoviridis [J], Chinese Journal of Experimental Formulas) and dissolved it in distilled water to prepare an oral preparation as a literature control group.

[0156] Test method: 162 C57BL / 6 mice, half male and half female. Randomly divided according to body weight: normal control group, model group, literature control group, Bai 1 drug group, Bai 6 drug group, Bai 7 drug group, Bai 8 drug group, Hong 1 drug group, Hong 6 drug group, Hong 7 drug group, Hong 8 drug group, Bai pure 2 drug group, Bai pure 3 drug group, Bai pure 6 drug group, Bai pure 7 drug group, Bai pure 10 drug group, Bai pure 11 drug group, Bai pure 14 drug group, Bai pure 15 drug group, Hong pure 2 drug group, Hong pure 3 drug group, Hong pure 6 drug group, Hong pure 7 drug group, Hong pure 10 drug group, Hong pure 11 drug group, Hong pure 14 drug group, Hong pure 15 drug group, a total of 27 groups, each group of 6 animals. Except for the normal control group, each group of mice was intraperitoneally injected with 100 mg / kg cytarabine once a day for 2 consecutive days, and from the 3rd day, intraperitoneally injected with 50 mg / kg cytarabine once a day for 3 consecutive days, and the total modeling time was 5 days. Each group was given corresponding drugs by gavage according to body weight, wherein each pure compound group was given a dose of 250 mg / kg, and each crude extract group was given a dose of 2000 mg / kg, once a day for 10 consecutive days. On the 10th day after modeling, 200 μL of blood was collected submandibularly, and the blood routine index changes were detected using a blood cell instrument.

[0157] Table 5. Platelet level changes of each group of mice after 10 days of modeling

[0158] Note: represents no significant difference (p>0.05) compared with the model control group; * represents p<0.05 compared with the model control group; ** represents p<0.01 compared with the model control group; *** represents p<0.001 compared with the model control group; * represents p<0.05 compared with the model control group; ** represents p<0.01 compared with the model control group; *** represents p<0.001 compared with the model control group;

[0159] Test results: On the 10th day after intraperitoneal injection of cytarabine in mice, the peripheral platelet count of the model group mice decreased significantly compared with that of normal mice, indicating that the chemotherapy drug-induced thrombocytopenia model was successfully constructed. Each purified component of Radermachia hancei and Radermachia sinensis showed a significant effect of improving the peripheral blood platelet level of mice compared with the model group, and compared with the peripheral platelet count of the literature control group, the purified components prepared by the application had more significant efficacy, especially the drug groups with relatively high total phenol content, which showed the most significant effect of promoting peripheral blood platelet production. In addition, each crude extract of Radermachia hancei and Radermachia sinensis also had a certain effect of increasing platelets, but required a higher dose. The results are shown in Table 5. The purified components of Radermachia hancei and Radermachia sinensis prepared by the application have a significant effect of promoting peripheral blood platelet production, and the efficacy is positively correlated with the total phenol content.

[0160] The above description is only the preferred 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 in 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. Use of traditional Chinese medicine Lysidice racemosa or Lysidice racemosa extract in preparation of a drug for blood system diseases.

2. Use according to claim 1, characterized in that The blood system diseases include, but are not limited to, thrombocytopenia, iron deficiency anemia, aplastic anemia, acute leukemia, idiopathic thrombocytopenic purpura (ITP), primary immune thrombocytopenia or chronic anemia; the blood system disease is preferably thrombocytopenia.

3. Use according to any one of claims 1-2, characterized in that The drug can promote peripheral platelet production or improve platelet activity.

4. Use according to any one of claims 1 to 2, characterized in that The traditional Chinese medicine Lysidice racemosa is selected from Lysidice fortunei and / or Lysidice rubescens; the Lysidice racemosa extract is selected from Lysidice fortunei extract and / or Lysidice rubescens extract.

5. Use according to any one of claims 1-2, characterized in that The Lysidice racemosa extract includes Lysidice racemosa crude extract and / or Lysidice racemosa purified extract.

6. Use according to claim 5, characterized in that The Lysidice racemosa crude extract is obtained by extracting the traditional Chinese medicine Lysidice racemosa with water or an alcohol aqueous solution.

7. Use according to claim 5, characterized in that The Lysidice racemosa crude extract is prepared by the following method: step (1), crushing Lysidice racemosa and adding water or an organic solvent as an extractant; step (2), reflux extraction and vacuum concentration to obtain the Lysidice racemosa crude extract.

8. Use according to any one of claims 6-7, characterized in that The Lysidice racemosa crude extract is prepared by the following method: step (1), crushing Lysidice racemosa, sieving, adding water or an alcohol organic solvent or an ester organic solvent or a ketone organic solvent as an extractant, and standing; step (2), reflux extraction, combining the extract, and vacuum concentration to obtain the Lysidice racemosa crude extract.

9. Use according to claim 8, characterized in that, In step (1), the ratio of Lysidice racemosa to the extractant is 1:1-12 kg / L, and the standing time is 0.5-18 h; in step (2), the reflux extraction is performed for 1-4 times, and the extraction time is 0.2-4 h each time. Preferably, in step (1), the sieving is 18 mesh; the ratio of Lysidice racemosa to the extractant is 1:2-10 kg / L, the standing time is 1-12 h, the reflux extraction is performed for 1-3 times, and the extraction time is 0.5-2 h each time; the extract is vacuum concentrated to a relative density of 1.10-1.30 (60°C).

10. Use according to any one of claims 7-8, characterized in that In step (1), the alcohol organic solvent is methanol, ethanol, propanol or butanol; the ester organic solvent is ethyl formate, ethyl acetate, propyl acetate, isopropyl acetate or butyl acetate; and the ketone organic solvent is acetone, butanone or pentanone. Preferably, the mass fraction of the alcohol organic solvent is 5%-100%, 10%-80%, 20%-60% or 40%-60%. Further preferably, the mass fraction of the alcohol organic solvent is 20%, 40%, 60%, 80% or 100%.

11. Use according to any one of claims 5 to 7, characterized in that The total phenol content in the Lysidice racemosa crude extract ranges from 1%-99%, 5%-95%, 6%-90%, 7%-85%, 8%-80%, 9%-78%, 10%-76%, 11%-74%, 12%-70%, 13%-68%, 14%-66%, 15%-64%, 16%-62%, 18%-60%, 20%-58%, 22%-56%, 24%-54%, 26%-52%, 28%-50%, 30%-48%, 32%-46%, 34%-44%, 32%-42%, 34%-40% or 36%-38%. Preferably, the total phenol content of the crude extract of the Magnoliae officinalis is 7.3%, 19.4%, 25.4%, 31.3%, 20.1%, 23.1%, 6.6%, 5.8%, 7.3%, 15.2%, 22.3%, 30.2%, 19.4%, 21.4%, 7.0%, 5.3%, 17.8%, 62.3%, 50.4%, 42.1%, 15.4%, 70.2%, 58.7%, 43.3%, 10.2%, 55.7%, 48.9%, 40.3%, 19.1%, 58.2%, 55.9%, or 38.9%.

12. The use according to claim 5, characterized in that The purified extract of Magnoliae officinalis is prepared by the following method: the crude extract of Magnoliae officinalis is purified by macroporous resin to obtain different polar parts of the purified extract of Magnoliae officinalis.

13. The use according to claim 5, characterized in that The purified extract of Magnoliae officinalis is prepared by the following method: the crude extract of Magnoliae officinalis is dissolved with distilled water, adsorbed by a macroporous resin column, eluted with deionized water, discarded, and then eluted with different concentrations of alcohol organic solvents. The eluate of different concentrations of alcohol organic solvents is collected, concentrated under reduced pressure, and dried to obtain the purified extract of Magnoliae officinalis. The macroporous resin column is independently selected from a macroporous resin column of type HPD-300, HPD-400, HPD-450, HPD-600, HPD-826, ADS-17, ADS-21, AB-8, D101, or D301.

14. The use according to any one of claims 12-13, wherein the macroporous resin column is selected from one of the following models: AB-8, HPD-300, ADS-17 or D101; 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, the loading amount is 10-60 mL, the static adsorption time is 2-24 h; the ionized water elution amount is 1BV-6BV, the alcohol organic solvent is ethanol, the mass fraction is 10%-80%, and the amount is 2-10BV. 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, a loading amount of 20-40 mL, and a static adsorption time of 4-24 h; the ionized water elution amount is 2-6 BV; the ethanol gradient elution mass fraction is 10%, 30%, 50%, or 70%, and the amount is 2-8 BV. More preferably, the adsorption flow rate is 5 mL·min -1 More preferably, the adsorption flow rate is 5 mL·min More preferably, the adsorption flow rate is 5 mL·min 15. The use according to claim 5, characterized in that The total phenol content of the purified extract of Magnoliae officinalis ranges from 1% to 99%, 5% to 95%, 6% to 90%, 7% to 85%, 8% to 80%, 9% to 78%, 10% to 76%, 11% to 74%, 12% to 70%, 13% to 68%, 14% to 66%, 15% to 64%, 16% to 62%, 18% to 60%, 20% to 58%, 22% to 56%, 24% to 54%, 26% to 52%, 28% to 50%, 30% to 48%, 32% to 46%, 34% to 44%, 32% to 42%, 34% to 40%, or 36% to 38%. Preferably, the total phenol content of the purified extract of Magnoliae officinalis is 17.8%, 62.3%, 50.4%, 42.1%, 15.4%, 70.2%, 58.7%, 43.3%, 10.2%, 55.7%, 48.9%, 40.3%, 19.1%, 58.2%, or 38.9%.

16. The use according to claim 1 or 2, wherein the medicament is a pharmaceutical composition comprising Magnoliae officinalis or an extract of Magnoliae officinalis.

17. The use according to claim 16, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

18. The use according to claim 17, wherein the pharmaceutical composition is prepared into any pharmaceutically acceptable dosage form.

19. The use according to claim 18, wherein the pharmaceutically acceptable dosage form is selected from tablets, capsules, solutions, granules, powders, ointments, pills, suspensions, suppositories, liniments, emulsions, smears, patches, or sprays.

20. The use according to claim 19, wherein the dosage form is in unit dosage form, each unit dosage contains 1-10 mg, or 10-100 mg, or 100-1000 mg, or 1000-10000 mg of traditional Chinese medicine Radermachia fraxinifolia Lindl. or Radermachia fraxinifolia Lindl. extract.

Images