CN103585259B - The application of a kind of Mucuna birdwoodiana Tutcher extract in preparation treatment AIDS-treating medicine - Google Patents
The application of a kind of Mucuna birdwoodiana Tutcher extract in preparation treatment AIDS-treating medicine Download PDFInfo
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Abstract
The present invention relates to a kind of novelty teabag of Mucuna birdwoodiana Tutcher extract, this novelty teabag is the application of Mucuna birdwoodiana Tutcher extract in preparation treatment AIDS-treating medicine.Described Mucuna birdwoodiana Tutcher extract is prepared as follows: after being extracted by Mucuna birdwoodiana, obtained by twice purification with macroreticular resin.Mucuna birdwoodiana Tutcher extract provided by the present invention is significant to preparation treatment AIDS-treating medicine.
Description
Technical field
The invention belongs to biomedicine field, relate to the application of a kind of Chinese medicinal material extract in preparation treatment AIDS-treating medicine, specifically relate to the application of a kind of Mucuna birdwoodiana Tutcher extract in preparation treatment AIDS-treating medicine.
Background technology
Acquired immune deficiency syndrome (AIDS) is the new virus sexually transmitted disease found in 1981, and the traditional Chinese medical science past is not on the books to it.The traditional Chinese medical science is dialectical to acquired immune deficiency syndrome (AIDS), thinks that acquired immune deficiency syndrome (AIDS) belongs to epidemic disease category.The cause of disease is the scorchingly hot heresy invasion and attack of epidemic disease, deficiency of vital QI and causing.Pathogenesis is mainly weakened defensive QI, and yang-energy damages, and internal organs are got involved, and causes that the deficiency of the lung is insufficiency of the spleen suffers from a deficiency of the kidney, secondary infection.Thermal burn venation, coagulation of QI-blood, Xing Cheng mass in the abdomen lump tumor.
Syndrome Differentiation of Chinese Medicine " card ": mainly distinguish ebb and flow between pathogen and vital QI and deficiency syndrome excess syndrome.HIV (human immunodeficiency virus) is evil poison, and the change of disease in evil poison enters, the dynamic change of just resisting with the heresy of body healthy energy is advancing of disease process.Infect early stage based on the domination of pathogen, mid-term " causes void because of real ", late period " excess caused by deficiency ".The acquired immune deficiency syndrome (AIDS) state of an illness copies changeable, and acquired immune deficiency syndrome (AIDS) and secondary infection thereof all can be formed " disease ", and has difference " type " and " phase ".The traditional Chinese medical science has dialectical differential diagnosis of diseases to acquired immune deficiency syndrome (AIDS), the typing different diagnosis and treatment project such as by stages.
The Chinese medicine of acquired immune deficiency syndrome (AIDS) still belongs to conceptual phase at present, at home and abroad there is no at present generally acknowledged curative effect certainly go through produce AIDS virus resisting Chinese medicine or Chinese medicine monomer compound.AIDS virus resisting plant amedica and Chinese medicine research mainly divide 3 kinds of approach, and the AIDS virus resisting Chinese medicinal plant medicine that smoking set obtains and composition thereof, some is in clinic trial.
Since HIV (human immunodeficiency virus) finds, use HIV (human immunodeficiency virus) cell culture and reverse transcriptase in the world, the ill vitro test method such as protease and intergrase, while extensive SCREENED COMPOUND, also plant and Chinese herbal medicine extract and composition thereof is screened in a large number, also studied some compound Chinese medicinal preparation, find the active drug of inhibiting HIV in vitro, the viremia of the suppressed simian immunodeficiency virus had (SIV) infected monkey.Some is in clinic trial, not yet clear and definite effect.
China removes Africa treatment AIDS patients from 1987 Nian Pai combination of Chinese and Western medicine medical teams.Domestic before 2002, lack specially good effect Anti-AIDS Drugs, Chinese and western medicine, combines with dialectical to AIDS patients differential diagnosis of diseases according to theory of Chinese medical science clinical, with heat-clearing and toxic substances removing, strengthening the body resistance, or removing heat from blood the kidney invigorating, the rule for the treatment of such as blood circulation promoting and blood stasis dispelling, by the property of Chinese materia medica, select Chinese medicine, prescribed treatment.By clinical practice, think that some Medical Complex Prescription in Treating AIDS can improve symptom, mitigate the disease.In recent years to the AIDS patients that laboratory diagnosis is made a definite diagnosis, adopt treatment by Chinese herbs, if matched group, at treatments period, periodic detection blood viral charge (HIV-RNA) and cd4 cell level, evaluation effect.There is report to improve symptom, improve immunity, mostly still in clinical verification.Enter clinical AntiHIV1 RT activity single medicinal material or plant amedica and Chinese medicine compound as follows:
Glycyrrhizin: glycyrrhizin (glycyrrhizin, GL) be the glycyrrhizin that glycyrrhiza uralensis fisch (Glycyrrhiza glabra L) extracts, there is broad-spectrum disease resistance toxic action, vitro inhibition HIV1-RT, HIV cell fusion is suppressed in cell culture, also resisiting influenza virus and SARS virus, the dead and pneumonopathy change of the pneumonia that experiment in vivo proves to protect influenza infection mice to cause; The CCl of protection mice
4toxic liver injury; Induce gamma interferon; There is parahormone effect.Japan's its intravenous administrations chronic hepatitis B conventional, also once tried out treatment AIDS patients, can suppress serum HIV-1-P24 antigen, raised cd4 cell.Lv Weibai professor 1988-1992 can treat AIDS patients African Tanzania application glycyrrhizin oral formulations gram Chinese mugwort can improve symptom, and cd4 cell slightly rises.Its effect is not yet verified.
Trichosanthin: trichosanthin (Trichosanthin, TCS, GLQ223, Q material) belong to Cucurbitaceae (Cucurbitaceae) for Chinese medicine Fructus Trichosanthis (Trichosanthes kirilowii), the main Chinese medicine ingredients of China's Chinese medicine compound for miscarrying among the people is Fructus Trichosanthis, the effective ingredient of China scientist isolation identification device antiearly pregnancy and induced labor is trichosanthin, molecular weight is 27kD, one, secondary structure and three dimensional structure are understood fully completely, for the toxalbumin of Profilin synthesis, ribosome inactivating protein (the Ribosome Inactivating Protein of strand, RIP).The U.S. is when screening anti-HIV-1 medicines, find that it suppresses HIV-1 cell fusion in lymphocyte is cultivated, thereafter find P24 antigen and the RNA of HIV-1 in mononuclear phagocyte is cultivated, blood plasma HIV-1 can be suppressed in the intravenous injection of 3 simian acquired immunodeficiency syndrome virus (SIV) infected monkeys.The U.S. uses intravenous administrations AIDS patients in forest farm, and HIV-1-P24 antigen in the T lymphocyte that peripheral blood HIV-1 can be suppressed to infect and mononuclear phagocyte, makes CD4/ CD8 ratio slightly raise.Due to anaphylactic reaction can be caused dead, neurovirulent Mental Subnormality and nephrotoxicity reaction, clinical experiment stops.From Radix Trichosanthis, isolate other albumen TAP29 in recent years, molecular weight, at 29 kD, also have the effect suppressing HIV-1, but toxicity is less, not in clinic trial.In addition the bitter melon protein of Fructus Momordicae charantiae (MOmordica charantia) and the Radix Phytolaccae albumen (Pokeweed protein) etc. of Radix Phytolaccae (Phytolacca americana) have the effect suppressing HIV.
Hypericin: plant Herba Hyperici Monogyni (St. John Wort, Hypericum perforatum) its extract for treating depression of folk tradition, the active component proposed from flower is hypericin (hypericin), it is the multi-ring dimerization anthraquinone analog compound of aromatic series, for anti-depression drug, complete synthesis.Hypericin has multiple antiviral activity, in cell culture, suppress HIV, and its mechanism of action is undressing of blocking virus, and assembling and release, the effect of killing the virus, by blood brain barrier.Murine leukemia virus can be suppressed in cell culture to breed.The splenomegaly that murine leukemia virus infecting mouse causes can be suppressed in vivo, duck hepatitis B virus infection Sanguis Anas domestica also can be suppressed to ask viral DNA, and have broad spectrum antibiotic active.Once in treatment on probation of clinical I/II phase HIV sufferers, test had been stopped because there being light sensitization.
1992 year report the native compound that from Malaysian rainforest plant Calophyllum lanigerum be separated dextrorotation lane element A: dextrorotation lane element A(+Calanolide A), Mus coumarin kind compound, suppress HIV1-RT, but invalid to HIV-2.Belong to non-nucleoside HIV1-RT inhibitor, in cell culture, multi-to-multi lives in the responsive and drug resistance HIV-1 of the AZT of clinical separation effectively, and and AZT, NVP have synergism.In HIV hollow fiber pipe Mice Body, prove there is AIDS virus resisting effect in experimental model (HIV hollow fiber mouse model), can drug effect be strengthened with AZT use in conjunction.Complete synthesis, having carried out its clinical trial of I-II proves there is the therapeutic effect suppressing HIV-1 to infect, and toxicity is little, and side effect is light, has headache and inappetence etc., effective with AZT drug combination.Be the single plant component uniquely entering II phase clinical experiment at present, observing in checking curative effect, not yet ratify to produce.
Herba Sidae Rhombifoliae soup: Herba Sidae Rhombifoliae soup (Radix Bupleuri, Radix Ginseng, the Rhizoma Pinelliae, Radix Scutellariae, Radix Glycyrrhizae, Rhizoma Zingiberis Recens, Fructus Jujubae)
It is Zhang Zhongjing Treatise on Febrile Diseases side.Hepatitis B is treated with it by Japan, finds that it suppresses HIV in cell culture.The U.S. is used for the treatment of numerical example AIDS patients clinical, can reduce blood-serum P 24 antigen.We prove that it has the effect suppressing HIV1-RT and suppress HIV-1 in cell culture.Study the effect of its 7 kinds of composition Chinese medicines respectively, find that only Radix Scutellariae water extract and composition baicalin (baicalin) thereof have and suppress HIV-1 effect, HIV1-RT is suppressed in vitro in cell free system, HIV-1 cytopathy is suppressed in human T-cell and peripheral blood lymphocytes cultivate, FLuorescent antigen and P24 antigen, in body, oral splenomegaly and the blood leukocytes of suppressing of murine leukemia virus infecting mouse raises.Japan Ono reports that noroxylin (baicalien) suppresses HIV1-RT.
In grind No. I: in grind No. I commodity such as (principal agent has Herba Violae, Radix Trichosanthis composition) and be called: Chinese mugwort
Gram electuary take heat-clearing and toxic substances removing as the rule for the treatment of, by Chinese medicine research institute of China prescription 1992-1993 in African Tanzania to the clinical practice of early metaphase acquired immune deficiency syndrome (AIDS), think to have and necessarily improve symptom, improve the effect of immunity.Laboratory proves that this medicine vitro inhibition hiv reverse transcriptase is active, simian acquired immunodeficiency syndrome poison infected monkey experimental therapy result is shown in body, plasma viral titre can be reduced, and cd4 cell number and CD4/CD8 ratio and T can be improved, bone-marrow-derived lymphocyte numerical value, inducement interferon and generation immunoloregulation function.Once in Africa and China's clinic trial, not yet clinical verification was ratified.
Zhongyan No. 2: Zhongyan No. 2 (Radix Astragali, Radix Ginseng, Radix Angelicae Sinensis, Fructus Lycii and Radix Glycyrrhizae etc.) is China's traditional Chinese medical science
Academy medical team prescription.1992 at African Tanzania treatment AIDS patients.1998-1999 is at African special outpatient clinic to 29 routine AIDS patients treatments, and 2/16 routine HIV virus load declines, and the routine CD4 of CD4 8/29 raises.2001-2002 helps the routine AIDS patients of peace hospitalize 15 in Beijing, 3/15 routine virus load declines, and the routine CD4 of CD4 1/15 raises.Be improved the effect of symptom.Experiment proves that this medicine shows simian acquired immunodeficiency syndrome poison infected monkey experimental therapy in human body, and medication 4-12 week can reduce plasma viral titre, P27 antigen, and can improve cd4 cell number, has activation to infected monkey lymphocyte.
Mucuna birdwoodiana is the dry seed of leguminous plant Mucuna birdwoodiana Mucuna birdwoodiana Tutcher.Collecting fruit, took out seed, dried autumn.Record in " Drug Standard of Ministry of Public Health of the Peoples Republic of China Tibetan medicine (first) ", standard number: WS3-BC-0030-95.Property is put down, sweet in the mouth.There is spleen-heat clearly, dredging collateral, the effect of keeping fit.For pneumonopathy, the splenopathy, meridians are sick, and Baconic is sick, nosotoxicosis, and external application is subsided a swelling.Kernel nourishes, and increases seminal fluid.The common drug of the minority areas such as illiteracy is hidden for China.But about the basic research of Mucuna birdwoodiana is still very limited, the follow-up promotion and application of this medical material are restricted.Modern study rarely has report about the chemical composition of Mucuna birdwoodiana and pharmacological action.
Domestic patent search result, has no Mucuna birdwoodiana Patents.
Above-mentioned document and patent etc., there is not yet Mucuna birdwoodiana or the Mucuna birdwoodiana Tutcher extract report for the preparation for the treatment of AIDS-treating medicine.
Summary of the invention
The object of the present invention is to provide the application of a kind of Mucuna birdwoodiana Tutcher extract in preparation treatment AIDS-treating medicine.
The present invention is achieved through the following technical solutions:
The present invention's Mucuna birdwoodiana used is the dry seed of leguminous plant Mucuna birdwoodiana Mucuna birdwoodiana Tutcher.
The application of a kind of Mucuna birdwoodiana Tutcher extract of the present invention in preparation treatment AIDS-treating medicine, the preparation method of described Mucuna birdwoodiana Tutcher extract is:
(1) Mucuna birdwoodiana, with concentration 50%-95% ethanol as solvent, extract at 30 DEG C of-60 DEG C of warm macerating, extraction time is 2-6 time, each extraction time is 2-8 hour, each solvent load is 10-30 times of Mucuna birdwoodiana weight, filters, obtains medicinal residues A and extracting solution, extracting solution reclaims ethanol, be concentrated into relative density d=1.05-1.12, filter, obtain medicinal liquid A;
(2) by the medicinal liquid A that step (1) obtains, by nonpolar macroporous adsorption resin, first wash with water, then use the alcoholic solution eluting of concentration 60%-95%, collect different concentration ethanol eluent, concentrate drying, obtains Mucuna birdwoodiana Tutcher extract A;
(3) step (1) being obtained medicinal residues A water is solvent, reflux, extract, and extraction time is 2-5 time, each extraction time is 1-6 hour, and each solvent load is 6-20 times of Mucuna birdwoodiana weight, filters, extracting solution is concentrated into relative density d=1.05-1.12, filters, obtains medical liquid B;
(4) by the medical liquid B that step (3) obtains, by polar macroporous adsorption resin, first wash with water, then use the alcoholic solution eluting of concentration 30%-80%, collect different concentration ethanol eluent, concentrate drying, obtains Mucuna birdwoodiana Tutcher extract B;
(5) by above-mentioned Mucuna birdwoodiana Tutcher extract A, Mucuna birdwoodiana Tutcher extract B, wherein one or both mix by a certain percentage, obtain Mucuna birdwoodiana Tutcher extract of the present invention.
The preparation method of Mucuna birdwoodiana Tutcher extract is:
(1) Mucuna birdwoodiana, with concentration 50%-95% ethanol as solvent, extract at 30 DEG C of-60 DEG C of warm macerating, extraction time is 2-6 time, each extraction time is 2-8 hour, each solvent load is 10-30 times of Mucuna birdwoodiana weight, filters, obtains medicinal residues A and extracting solution, extracting solution reclaims ethanol, be concentrated into relative density d=1.05-1.12, filter, obtain medicinal liquid A;
(2) by the medicinal liquid A that step (1) obtains, by nonpolar macroporous adsorption resin, first wash with water, then use the alcoholic solution eluting of concentration 60%-95%, collect different concentration ethanol eluent, concentrate drying, obtains Mucuna birdwoodiana Tutcher extract A;
(3) step (1) being obtained medicinal residues A water is solvent, reflux, extract, and extraction time is 2-5 time, each extraction time is 1-6 hour, and each solvent load is 6-20 times of Mucuna birdwoodiana weight, filters, extracting solution is concentrated into relative density d=1.05-1.12, filters, obtains medical liquid B;
(4) by the medical liquid B that step (3) obtains, by polar macroporous adsorption resin, first wash with water, then use the alcoholic solution eluting of concentration 30%-80%, collect different concentration ethanol eluent, concentrate drying, obtains Mucuna birdwoodiana Tutcher extract B;
(5) above-mentioned Mucuna birdwoodiana Tutcher extract A, Mucuna birdwoodiana Tutcher extract B are mixed, obtain Mucuna birdwoodiana Tutcher extract of the present invention.
The preparation method of preferred Mucuna birdwoodiana Tutcher extract is:
(1) Mucuna birdwoodiana, with concentration 70% ethanol as solvent, extract at 50 DEG C of warm macerating, extraction time is 3 times, each extraction time is 4 hours, each solvent load is 20 times of Mucuna birdwoodiana weight, filters, obtains medicinal residues A and extracting solution, extracting solution reclaims ethanol, be concentrated into relative density d=1.10, filter, obtain medicinal liquid A;
(2) by the medicinal liquid A that step (1) obtains, by D101 macroporous adsorbent resin, first wash with water, then use the alcoholic solution eluting of concentration 60%, collect 60% concentration ethanol eluent, concentrate drying, obtains Mucuna birdwoodiana Tutcher extract A;
(3) step (1) being obtained medicinal residues A water is solvent, reflux, extract, and extraction time is 3 times, each extraction time is 2 hours, and each solvent load is 10 times of Mucuna birdwoodiana weight, filters, extracting solution is concentrated into relative density d=1.11, filters, obtains medical liquid B;
(4) by the medical liquid B that step (3) obtains, by DM130 macroporous adsorbent resin, first wash with water, then use the alcoholic solution eluting of concentration 70%, collect 70% concentration ethanol eluent, concentrate drying, obtains Mucuna birdwoodiana Tutcher extract B;
(5) above-mentioned Mucuna birdwoodiana Tutcher extract A, Mucuna birdwoodiana Tutcher extract B are mixed, obtain Mucuna birdwoodiana Tutcher extract of the present invention.
The preparation method of Mucuna birdwoodiana Tutcher extract of the present invention, is characterized in that: the nonpolar macroporous adsorption resin adopted is AB-8 macroporous adsorbent resin, D101 macroporous adsorbent resin; The polar macroporous adsorption resin adopted is DM130 macroporous adsorbent resin, LSA-40 macroporous adsorbent resin.
Mucuna birdwoodiana Tutcher extract of the present invention is for the preparation for the treatment of AIDS-treating medicine.
Mucuna birdwoodiana Tutcher extract A of the present invention, the Mucuna birdwoodiana Tutcher extract B application in preparation treatment AIDS-treating medicine.
The treatment AIDS-treating medicine that Mucuna birdwoodiana Tutcher extract of the present invention and chemical drugs or Chinese medicine or natural drug form.
The treatment AIDS-treating medicine that Mucuna birdwoodiana Tutcher extract A of the present invention, Mucuna birdwoodiana Tutcher extract B and chemical drugs or Chinese medicine or natural drug form.
Mucuna birdwoodiana Tutcher extract of the present invention, by adding the various adjuvants that pharmaceutics allows, makes the peroral dosage forms such as the tablet on pharmaceutics, granule, capsule.
The present invention's exploratory development first treats acquired immune deficiency syndrome (AIDS) extract with the dry seed of leguminous plant Mucuna birdwoodiana Mucuna birdwoodiana Tutcher for raw material extracts preparation.This experimentation shows, the external infected by influenza of each extract of Mucuna birdwoodiana, parainfluenza virus I type (HVJ), respiratory syncytial virus (RSV), Coxsackie virus B 4 type (C
oxb
4), adenovirus type III ((Ard
3) and rhinoviral cytopathy comparatively virus control group alleviate all to some extent.There is obvious inhibitory action to mouse influenza pneumonia in body, have significant protective effect to influenza to dead mouse, show that each extract of Mucuna birdwoodiana has good antivirus action.Clinical observation result of the test shows, takes 18 routine HIV sufferers of Mucuna birdwoodiana Tutcher extract treatment, obviously improves symptom, repairs hepatic and renal function, improve, stablize or prolong CD
4decline, suppress virus replication etc. to have obvious curative effect.
Detailed description of the invention
Below by specific experiment example and embodiment, the application of a kind of Mucuna birdwoodiana Tutcher extract in preparation treatment AIDS-treating medicine is described further, but is not limited to the present invention.
Embodiment 1: the preparation of Mucuna birdwoodiana Tutcher extract
(1) Mucuna birdwoodiana 15kg, with concentration 70% ethanol as solvent, extract at 50 DEG C of warm macerating, extraction time is 3 times, each extraction time is 4 hours, each solvent load is 20 times of Mucuna birdwoodiana weight, filters, obtains medicinal residues A and extracting solution, extracting solution reclaims ethanol, be concentrated into relative density d=1.10, filter, obtain medicinal liquid A;
(2) by the medicinal liquid A that step (1) obtains, by D101 macroporous adsorbent resin, first wash with water, then use the alcoholic solution eluting of concentration 60%, collect 60% concentration ethanol eluent, concentrate drying, obtains Mucuna birdwoodiana Tutcher extract A;
(3) step (1) being obtained medicinal residues A water is solvent, reflux, extract, and extraction time is 3 times, each extraction time is 2 hours, and each solvent load is 10 times of Mucuna birdwoodiana weight, filters, extracting solution is concentrated into relative density d=1.11, filters, obtains medical liquid B;
(4) by the medical liquid B that step (3) obtains, by DM130 macroporous adsorbent resin, first wash with water, then use the alcoholic solution eluting of concentration 70%, collect 70% concentration ethanol eluent, concentrate drying, obtains Mucuna birdwoodiana Tutcher extract B;
(5) above-mentioned Mucuna birdwoodiana Tutcher extract A, Mucuna birdwoodiana Tutcher extract B are mixed, obtain Mucuna birdwoodiana Tutcher extract of the present invention.
Embodiment 2: the preparation of Mucuna birdwoodiana Tutcher extract
(1) Mucuna birdwoodiana 20kg, with concentration 50% ethanol as solvent, extract at 60 DEG C of warm macerating, extraction time is 6 times, each extraction time is 2 hours, each solvent load is 10 times of Mucuna birdwoodiana weight, filters, obtains medicinal residues A and extracting solution, extracting solution reclaims ethanol, be concentrated into relative density d=1.12, filter, obtain medicinal liquid A;
(2) by the medicinal liquid A that step (1) obtains, by AB-8 nonpolar macroporous adsorption resin, first wash with water, then use the alcoholic solution eluting of concentration 60%, collect 60% concentration ethanol eluent, concentrate drying, obtains Mucuna birdwoodiana Tutcher extract A;
(3) step (1) being obtained medicinal residues A water is solvent, reflux, extract, and extraction time is 2 times, each extraction time is 6 hours, and each solvent load is 20 times of Mucuna birdwoodiana weight, filters, extracting solution is concentrated into relative density d=1.05, filters, obtains medical liquid B;
(4) by the medical liquid B that step (3) obtains, by LSA-40 polar macroporous adsorption resin, first wash with water, then use the alcoholic solution eluting of concentration 30%, collect 30% concentration ethanol eluent, concentrate drying, obtains Mucuna birdwoodiana Tutcher extract B;
(5) above-mentioned Mucuna birdwoodiana Tutcher extract A, Mucuna birdwoodiana Tutcher extract B are mixed, obtain Mucuna birdwoodiana Tutcher extract of the present invention.
Embodiment 3: the preparation of Mucuna birdwoodiana Tutcher extract
(1) Mucuna birdwoodiana, with concentration 95% ethanol as solvent, extract at 30 DEG C of warm macerating, extraction time is 2 times, each extraction time is 8 hours, each solvent load is 30 times of Mucuna birdwoodiana weight, filters, obtains medicinal residues A and extracting solution, extracting solution reclaims ethanol, be concentrated into relative density d=1.05, filter, obtain medicinal liquid A;
(2) by the medicinal liquid A that step (1) obtains, by D101 nonpolar macroporous adsorption resin, first wash with water, then use the alcoholic solution eluting of concentration 95%, collect 95% concentration ethanol eluent, concentrate drying, obtains Mucuna birdwoodiana Tutcher extract A;
(3) step (1) being obtained medicinal residues A water is solvent, reflux, extract, and extraction time is 5 times, and each extraction time is 1 hour, and each solvent load is 6 times of Mucuna birdwoodiana weight, and filter, extracting solution is concentrated into relative density d=1.12, filters, obtains medical liquid B;
(4) by the medical liquid B that step (3) obtains, by LSA-40 polar macroporous adsorption resin, first wash with water, then use the alcoholic solution eluting of concentration 80%, collect 80% concentration ethanol eluent, concentrate drying, obtains Mucuna birdwoodiana Tutcher extract B;
(5) above-mentioned Mucuna birdwoodiana Tutcher extract A, Mucuna birdwoodiana Tutcher extract B are mixed, obtain Mucuna birdwoodiana Tutcher extract of the present invention.
Embodiment 4: the preparation of Mucuna birdwoodiana Tutcher extract sheet
Example 1 Mucuna birdwoodiana Tutcher extract 322g, adds starch 70g, mixing, granulates, dry, sieves, adds microcrystalline Cellulose 25g, magnesium stearate 4.5g, and mixing, is pressed into 1000, obtains Mucuna birdwoodiana Tutcher extract sheet.
Embodiment 5: the preparation of Mucuna birdwoodiana Tutcher extract granule
Example 2 Mucuna birdwoodiana Tutcher extract 245g, adds dextrin 98g, mixing, granulates, and dry, granulate, obtains Mucuna birdwoodiana Tutcher extract granule.
Embodiment 6: the preparation of Mucuna birdwoodiana Tutcher extract capsule
Example 3 Mucuna birdwoodiana Tutcher extract 285g, adds starch 75g, mixing, granulates, and dry, granulate, adds magnesium stearate 3g, and mixing, obtains Mucuna birdwoodiana Tutcher extract capsule by encapsulated 1000.
Experimental example l: the antivirus action of each extract of Mucuna birdwoodiana
Test material: Mucuna birdwoodiana Tutcher extract, Mucuna birdwoodiana Tutcher extract A, Mucuna birdwoodiana Tutcher extract B(prepare by the embodiment of the present invention 1 method, and lot number is respectively: 20110808,20110809,20110810); Virazole (ribavirin) is produced by Hubei Province's institute of Pharmaceutical Industry, lot number: 20110503; SHUANGHUANGLIAN JIAONANG is produced by Jinghai Pharmaceutical Co., Ltd., Beijing, lot number: 20110701; Influenza virus A 3 type, the sick 4 type (C of coxsackie B race
oxb
4), parainfluenza virus I type (HVJ), respiratory syncytial virus (RSV), adenovirus type III (Ard
3), rhinovirus 14 type and influenza virus A-prime Mus lung adapted strain FM1, respectively purchased from Virology Inst., Chinese Academy of Preventive Medical Science and institute of pediatrics; People's laryngeal carcinoma passage cell HEP-2 strain, purchased from Ministry of Public Health pharmaceutical biological product calibrating institute; Containing 10% calf serum, 0.29mg paddy ammonia phthalein amine/m1,100 μ l are blue or green, the Eagle'MEM of Streptomycin; Cell maintenance medium, except contained calf serum is except 2%, all same culture fluid of other content.Kunming mouse, is provided by Shandong Province's Experimental Animal Center.
Experimental technique
1. antiviral effect in vitro: the toxicity test to Hep-2 cultured cell: medicinal liquid Eagle's is cultivated and makes 1:20-1:1280 doubling dilution, be added to and grow up in the HEP-2 Tissue Culture Plate of monolayer, 100 μ l/ holes, each dilution factor medicinal liquid does 3 multiple holes, establishes normal cell controls simultaneously.Culture plate is put 37 DEG C, 5% C0
2cultivate 4 days in incubator, observation of cell growing state under every day inverted microscope, determines that cell does not occur that the minimum of obvious regression releases rare multiple, postpones and accomplish minimum effective drug concentration (maximum dilution multiple) during experiment.50% toxic concentration (TC is calculated by Reed-Muench method
50) and maximal non-toxic concentration (TC
0), the results are shown in Table 1.
The TC of table 1 pair cultured cell
0tC
50(mg/ml).
2. interior resisting virus effect: the inhibitory action to mouse influenza virus pneumonia: get mice 70,7 groups are divided at random by body weight, be respectively intact animal's matched group, virus-infected controls group, trial drug 3 groups (Mucuna birdwoodiana Tutcher extract, Mucuna birdwoodiana Tutcher extract A, Mucuna birdwoodiana Tutcher extract B), virazole group, SHUANGHUANGLIAN JIAONANG group.Except Normal group, mice is used ether light anesthesia, infect with 15 LD50 influenza virus drop noses, every 0.05ml.Gastric infusion from infecting the previous day, every day 3 times, each 0.5ml, continuous 5 days, matched group is distilled water gavage under equal conditions.Dissect after within 6th day, taking Mouse Weight, perusal pulmonary lesion, the degree of record pulmonary liver sample consolidation, wins full lung and weighs, calculate Lung Exponent value one by one, and obtain lung index.Result: the mice pneumonia that Mucuna birdwoodiana each extract infected by influenza causes all has obvious inhibitory action, the results are shown in Table 2.
The inhibitory action of table 2 pair mouse influenza virus pneumonia
Note: compare with Normal group:
##p<0.01; Compare with virus control group: * * P<0.01, * P<0.05.
3, the protective effect of infected by influenza induced mice death: get mice 120, be divided into 6 groups at random by body weight.Be respectively trial drug three dosage groups, virazole group, SHUANGHUANLIAN group and virus-infected controls group.Each group of mice, with after ether light anesthesia, infects with 2 LD50 influenza virus drop noses, every 0.05ml.Gastric infusion from infecting the previous day, every day 3 times, each 0.5ml, continuous 7 days.The accumulative death toll infecting rear mice, calculates mortality rate, protective rate and increase in life span, the results are shown in Table 3,4.
Table 3 infected by influenza causes the protective effect of dead mouse
Note: compare with infection matched group:
*p<0.0l,
*p<0.05.
Table 4 infected by influenza causes the protective effect of dead mouse
Note: compare with infection matched group:
*p<0.0l,
*p<0.05.
Table 3; table 4 result show: mouse infection virus latter 14 days in; the death toll of Mucuna birdwoodiana each extract group mice comparatively infects matched group and obviously reduces; mice Average Survival natural law comparatively infects matched group and obviously extends, and shows that trial drug infected by influenza infection when this dosage causes dead mouse and has certain protective role.
The external infected by influenza of each extract of Mucuna birdwoodiana, parainfluenza virus I type (HVJ), respiratory syncytial virus (RSV), Coxsackie virus B 4 type (C is found in experimentation
oxb
4), adenovirus type III ((Ard
3) and rhinoviral cytopathy comparatively virus control group alleviate all to some extent.There is obvious inhibitory action to mouse influenza pneumonia in body, have significant protective effect to influenza to dead mouse, show that each extract of Mucuna birdwoodiana has good antivirus action, therefore have the basis developed preferably.
Experimental example 2: the clinical observation on the therapeutic effect of Mucuna birdwoodiana Tutcher extract treatment acquired immune deficiency syndrome (AIDS)
Physical data: 18 routine cases are all HIV sufferers through hospital diagnosis, accept clinical research voluntarily; Can adhere to that accepting assessment detects and treatment; Age 20-62 year; Man 11 example, female 7 example; 33 years old mean age.All case never received antiviral treatments.
Diagnostic criteria: all cases all meet HIV/AIDS diagnostic criteria.
Therapeutic Method: all oral Mucuna birdwoodiana Tutcher extract of the present invention of all cases, a 1g, 3 times on the one 30 days is a course for the treatment of.
Clinical efficacy evaluation criteria: " the Treating AIDS with Traditional Chinese Medicine pilot project clinical technology scheme " formulated according to State Administration of Traditional Chinese Medicine, the standard of curative effect evaluation is as follows: effectively: Clinical symptom and sign improves obviously, and total mark declines>=1/3; CD
4rising>=30% or 50 μ l; Virus load reduces >0.5log/ml; Stable: Clinical symptom and sign makes moderate progress, total mark decline <1/3; CD
4raise or decline <30% or 50 μ l; Virus load rises or decline <0.5log/ml.Invalid: Clinical symptom and sign is without improvement or increase the weight of, and total mark does not decline or increases; CD
4decline>=30% or 50 μ l; Viral load levels continues to rise, copy number rising >0.5log/ml.
Result is tired out in clinical sight: symptom and sign change before and after treatment: effective 13 examples, stable: 5 examples, invalid: 0 example.
CD before and after treatment
4cellular change: effective 15 examples is stable: 1 example, invalid: 2 examples.
Virus load change before and after treatment: effective 10 examples, stable: 2 examples, invalid: 6 examples.
Clinical observation result shows: the 18 routine HIV sufferers taking Mucuna birdwoodiana Tutcher extract treatment, obviously improves symptom, repairs hepatic and renal function, improve, stablize or prolong CD
4decline, suppress virus replication etc. to have obvious curative effect.
Claims (3)
1. the application of Mucuna birdwoodiana Tutcher extract in preparation treatment AIDS-treating medicine, the preparation method of described Mucuna birdwoodiana Tutcher extract is:
(1) Mucuna birdwoodiana, with concentration 50%-95% ethanol as solvent, extract at 30 DEG C of-60 DEG C of warm macerating, extraction time is 2-6 time, each extraction time is 2-8 hour, each solvent load is 10-30 times of Mucuna birdwoodiana weight, filters, obtains medicinal residues A and extracting solution, extracting solution reclaims ethanol, be concentrated into relative density 1.05-1.12, filter, obtain medicinal liquid A;
(2) by the medicinal liquid A that step (1) obtains, by nonpolar macroporous adsorption resin, first wash with water, then use the alcoholic solution eluting of concentration 60%-95%, collect different concentration ethanol eluent, concentrate drying, obtains Mucuna birdwoodiana Tutcher extract A;
(3) step (1) being obtained medicinal residues A water is solvent, reflux, extract, and extraction time is 2-5 time, each extraction time is 1-6 hour, and each solvent load is 6-20 times of Mucuna birdwoodiana weight, filters, extracting solution is concentrated into relative density 1.05-1.12, filters, obtains medical liquid B;
(4) by the medical liquid B that step (3) obtains, by polar macroporous adsorption resin, first wash with water, then use the alcoholic solution eluting of concentration 30%-80%, collect different concentration ethanol eluent, concentrate drying, obtains Mucuna birdwoodiana Tutcher extract B;
(5) above-mentioned Mucuna birdwoodiana Tutcher extract A, Mucuna birdwoodiana Tutcher extract B are mixed, obtain Mucuna birdwoodiana Tutcher extract.
2. application according to claim 1, is characterized in that the preparation method of described Mucuna birdwoodiana Tutcher extract is:
(1) Mucuna birdwoodiana, with concentration 70% ethanol as solvent, extract at 50 DEG C of warm macerating, extraction time is 3 times, each extraction time is 4 hours, each solvent load is 20 times of Mucuna birdwoodiana weight, filters, obtains medicinal residues A and extracting solution, extracting solution reclaims ethanol, be concentrated into relative density 1.10, filter, obtain medicinal liquid A;
(2) by the medicinal liquid A that step (1) obtains, by D101 macroporous adsorbent resin, first wash with water, then use the alcoholic solution eluting of concentration 60%, collect 60% concentration ethanol eluent, concentrate drying, obtains Mucuna birdwoodiana Tutcher extract A;
(3) step (1) being obtained medicinal residues A water is solvent, reflux, extract, and extraction time is 3 times, and each extraction time is 2 hours, and each solvent load is 10 times of Mucuna birdwoodiana weight, and filter, extracting solution is concentrated into relative density 1.11, filters, obtains medical liquid B;
(4) by the medical liquid B that step (3) obtains, by DM130 macroporous adsorbent resin, first wash with water, then use the alcoholic solution eluting of concentration 70%, collect 70% concentration ethanol eluent, concentrate drying, obtains Mucuna birdwoodiana Tutcher extract B;
(5) above-mentioned Mucuna birdwoodiana Tutcher extract A, Mucuna birdwoodiana Tutcher extract B are mixed, obtain Mucuna birdwoodiana Tutcher extract.
3. application according to claim 1, is characterized in that the treatment AIDS-treating medicine that described Mucuna birdwoodiana Tutcher extract and chemical drugs or Chinese medicine or natural drug form.
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| CN101537060A (en) * | 2009-04-29 | 2009-09-23 | 常俊平 | Medicine for treating AIDS |
| CN101756956A (en) * | 2009-03-25 | 2010-06-30 | 赵亚力 | Use of catechin for preparing retrovirus integrase inhibitors |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101756956A (en) * | 2009-03-25 | 2010-06-30 | 赵亚力 | Use of catechin for preparing retrovirus integrase inhibitors |
| CN101537060A (en) * | 2009-04-29 | 2009-09-23 | 常俊平 | Medicine for treating AIDS |
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