CN103585188B - The application of wall tapestry extract in preparation treatment AIDS-treating medicine - Google Patents
The application of wall tapestry extract in preparation treatment AIDS-treating medicine Download PDFInfo
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- CN103585188B CN103585188B CN201310525371.7A CN201310525371A CN103585188B CN 103585188 B CN103585188 B CN 103585188B CN 201310525371 A CN201310525371 A CN 201310525371A CN 103585188 B CN103585188 B CN 103585188B
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Abstract
The present invention relates to a kind of novelty teabag of wall tapestry extract, this novelty teabag is the application of wall tapestry extract in preparation treatment AIDS-treating medicine.Described wall tapestry extract is prepared as follows: after being extracted by wall tapestry, obtained by twice purification with macroreticular resin.Wall tapestry 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 wall tapestry 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.
Wall tapestry is the dry thallus of lichens Teloshistaceae plant wall tapestry Theloschestes flavicans (Sw.) Norm.After collection, removing earth, dries.Record in " Drug Standard of Ministry of Public Health of the Peoples Republic of China Tibetan medicine (first) ", standard number: WS3-BC-0125-95.Cool in nature, bitter in the mouth.There is heat extraction, effect of removing toxic substances.For lung-heat, liver-heat, arteries and veins heat, scorchingly hot.The common drug of the minority areas such as illiteracy is hidden for China.But about the basic research of wall tapestry is still very limited, the follow-up promotion and application of this medical material are restricted.Relevant wall tapestry chemical composition and pharmacological research have no report.
Domestic patent search result, has no wall tapestry Patents.
Above-mentioned document and patent etc., there is not yet wall tapestry or the wall tapestry 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 wall tapestry extract in preparation treatment AIDS-treating medicine.
The present invention is achieved through the following technical solutions:
The present invention's wall tapestry used is the dry thallus of lichens Teloshistaceae plant wall tapestry Theloschestes flavicans (Sw.) Norm.
The application of wall tapestry extract of the present invention in preparation treatment AIDS-treating medicine, the preparation method of described wall tapestry extract is:
(1) wall tapestry, with concentration 80%-100% ethanol as solvent, extract at 20 DEG C of-70 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 wall tapestry weight, filters, obtains medicinal residues A and extracting solution, extracting solution reclaims ethanol, concentrated, dry, obtain wall tapestry extract A;
(2) step (1) being obtained medicinal residues A concentration 0-50% ethanol is solvent, extract at 30 DEG C of-80 DEG C of warm macerating, extraction time is 2-5 time, each extraction time is 1-6 hour, and each solvent load is 6-20 times of wall tapestry weight, filters, extracting solution reclaims ethanol, be concentrated into relative density d=1.05-1.12, filter, obtain medical liquid B;
(3) by medical liquid B that step (2) obtains, pass through nonpolarity macroporous adsorptive resins chromatographic column, first wash with water, water elution liquid is directly by polar macroporous resin column, use the alcoholic solution eluting nonpolarity macroporous adsorptive resins chromatographic column of concentration 60%-95% again, collect different concentration ethanol eluent, concentrate drying, obtains wall tapestry extract B; Use the alcoholic solution eluting polar macroporous adsorbent resin column of concentration 60%-85% again, collect different concentration ethanol eluent, concentrate drying, obtains wall tapestry extract C;
(4) above-mentioned wall tapestry extract A, wall tapestry extract B, wall tapestry extract C, wherein one or both or three kinds mix by a certain percentage, obtain wall tapestry extract of the present invention.
The preparation method of wall tapestry extract is:
(1) wall tapestry, with concentration 80%-100% ethanol as solvent, extract at 20 DEG C of-70 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 wall tapestry weight, filters, obtains medicinal residues A and extracting solution, extracting solution reclaims ethanol, concentrated, dry, obtain wall tapestry extract A;
(2) step (1) being obtained medicinal residues A concentration 0-50% ethanol is solvent, extract at 30 DEG C of-80 DEG C of warm macerating, extraction time is 2-5 time, each extraction time is 1-6 hour, and each solvent load is 6-20 times of wall tapestry weight, filters, extracting solution reclaims ethanol, be concentrated into relative density d=1.05-1.12, filter, obtain medical liquid B;
(3) by medical liquid B that step (2) obtains, pass through nonpolarity macroporous adsorptive resins chromatographic column, first wash with water, water elution liquid is directly by polar macroporous resin column, use the alcoholic solution eluting nonpolarity macroporous adsorptive resins chromatographic column of concentration 60%-95% again, collect different concentration ethanol eluent, concentrate drying, obtains wall tapestry extract B; Use the alcoholic solution eluting polar macroporous adsorbent resin column of concentration 60%-85% again, collect different concentration ethanol eluent, concentrate drying, obtains wall tapestry extract C;
(4) by above-mentioned wall tapestry extract A, wall tapestry extract B, the mixing of wall tapestry extract C, wall tapestry extract of the present invention is obtained.
The preparation method of preferred wall tapestry extract of the present invention is:
(1) wall tapestry, with concentration 90% ethanol as solvent, extract at 50 DEG C of warm macerating, extraction time is 4 times, each extraction time is 4 hours, and each solvent load is 20 times of wall tapestry weight, filters, obtains medicinal residues A and extracting solution, extracting solution reclaims ethanol, concentrated, dry, obtains wall tapestry extract A;
(2) step (1) being obtained medicinal residues A concentration 30% ethanol is solvent, and extract at 70 DEG C of warm macerating, extraction time is 3 times, and each extraction time is 3 hours, each solvent load is 15 times of wall tapestry weight, filters, and extracting solution reclaims ethanol, be concentrated into relative density d=1.12, filter, obtain medical liquid B;
(3) by medical liquid B that step (2) obtains, by AB-8 nonpolarity macroporous adsorptive resins chromatographic column, first wash with water, water elution liquid is directly by the polar macroporous resin column of DM130, use the alcoholic solution eluting AB-8 nonpolarity macroporous adsorptive resins chromatographic column of concentration 85% again, collect 85% concentration ethanol eluent, concentrate drying, obtain wall tapestry extract B; Use the alcoholic solution eluting DM130 polar macroporous adsorbent resin column of concentration 70% again, collect 70% concentration ethanol eluent, concentrate drying, obtain wall tapestry extract C;
(4) by above-mentioned wall tapestry extract A, wall tapestry extract B, the mixing of wall tapestry extract C, wall tapestry extract of the present invention is obtained.
The preparation method of wall tapestry 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-10 macroporous adsorbent resin.
The application in preparation treatment AIDS-treating medicine of wall tapestry extract A of the present invention, wall tapestry extract B, wall tapestry extract C.
The treatment AIDS-treating medicine that wall tapestry extract of the present invention and chemical drugs or Chinese medicine or natural drug form.
The treatment AIDS-treating medicine that wall tapestry extract A of the present invention, wall tapestry extract B, wall tapestry extract C and chemical drugs or Chinese medicine or natural drug form.
Wall tapestry 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 is first that raw material extracts preparation treatment acquired immune deficiency syndrome (AIDS) extract with the dry thallus of lichens Teloshistaceae plant wall tapestry Theloschestes flavicans (Sw.) Norm.This experimentation shows, wall tapestry extract of the present invention can be used for preparation treatment AIDS-treating medicine, the splenomegaly that wall tapestry extract oral can suppress FLV to cause.Spleen is the first target organs of virus, and the spleen of FLV infected mice increases weight large I quantitative response virus quantity, and therefore can medicine suppress virus, and spleen index is a directly perceived and index reliably.Spleen has abundant lymphocyte and macrophage, and they all participate in humoral immune reaction, so wall tapestry extract can suppress splenomegaly in an experiment, shows it and not only has anti-FLV effect in body, also may have the effect improving humoral immune function.From the experimental result of amynologic index, wall tapestry extract has good rise effect to cellular immunization, particularly when the body's immunity that retrovirus causes is low, performance is particularly remarkable, showing this medicine can enhancing body cellular immune function, suppressing the immunodeficiency that FLV causes, is a good immunomodulator.
Detailed description of the invention
Below by specific experiment example and embodiment, the application of wall tapestry extract in preparation treatment AIDS-treating medicine is described further, but is not limited to the present invention.
Embodiment 1: the preparation of wall tapestry extract
(1) wall tapestry 15kg, with concentration 90% ethanol as solvent, extract at 50 DEG C of warm macerating, extraction time is 4 times, each extraction time is 4 hours, and each solvent load is 20 times of wall tapestry weight, filters, obtains medicinal residues A and extracting solution, extracting solution reclaims ethanol, concentrated, dry, obtains wall tapestry extract A;
(2) step (1) being obtained medicinal residues A concentration 30% ethanol is solvent, and extract at 70 DEG C of warm macerating, extraction time is 3 times, and each extraction time is 3 hours, each solvent load is 15 times of wall tapestry weight, filters, and extracting solution reclaims ethanol, be concentrated into relative density d=1.12, filter, obtain medical liquid B;
(3) by medical liquid B that step (2) obtains, by AB-8 nonpolarity macroporous adsorptive resins chromatographic column, first wash with water, water elution liquid is directly by the polar macroporous resin column of DM130, use the alcoholic solution eluting AB-8 nonpolarity macroporous adsorptive resins chromatographic column of concentration 85% again, collect 85% concentration ethanol eluent, concentrate drying, obtain wall tapestry extract B; Use the alcoholic solution eluting DM130 polar macroporous adsorbent resin column of concentration 70% again, collect 70% concentration ethanol eluent, concentrate drying, obtain wall tapestry extract C;
(4) by above-mentioned wall tapestry extract A, wall tapestry extract B, the mixing of wall tapestry extract C, wall tapestry extract of the present invention is obtained.
Embodiment 2: the preparation of wall tapestry extract
(1) wall tapestry 18kg, with concentration 80% ethanol as solvent, extract at 70 DEG C of warm macerating, extraction time is 2 times, each extraction time is 8 hours, and each solvent load is 10 times of wall tapestry weight, filters, obtains medicinal residues A and extracting solution, extracting solution reclaims ethanol, concentrated, dry, obtains wall tapestry extract A;
(2) step (1) being obtained medicinal residues A water is solvent, and extract at 80 DEG C of warm macerating, extraction time is 2 times, and each extraction time is 6 hours, each solvent load is 20 times of wall tapestry weight, filters, and extracting solution reclaims ethanol, be concentrated into relative density d=1.12, filter, obtain medical liquid B;
(3) by medical liquid B that step (2) obtains, by D101 nonpolarity macroporous adsorptive resins chromatographic column, first wash with water, water elution liquid is directly by the polar macroporous resin column of LSA-10, use the alcoholic solution eluting D101 nonpolarity macroporous adsorptive resins chromatographic column of concentration 95% again, collect 95% concentration ethanol eluent, concentrate drying, obtain wall tapestry extract B; Use the alcoholic solution eluting LSA-10 polar macroporous adsorbent resin column of concentration 60% again, collect 60% concentration ethanol eluent, concentrate drying, obtain wall tapestry extract C;
(4) by above-mentioned wall tapestry extract A, wall tapestry extract B/wall tapestry extract C mixing, wall tapestry extract of the present invention is obtained.
Embodiment 3: the preparation of wall tapestry extract
(1) wall tapestry 20kg, with 100% ethanol as solvent, extract at 20 DEG C of warm macerating, extraction time is 6 times, each extraction time is 2 hours, and each solvent load is 30 times of wall tapestry weight, filters, obtains medicinal residues A and extracting solution, extracting solution reclaims ethanol, concentrated, dry, obtains wall tapestry extract A;
(2) step (1) being obtained medicinal residues A concentration 50% ethanol is solvent, and extract at 30 DEG C of warm macerating, extraction time is 5 times, and each extraction time is 1 hour, each solvent load is 6 times of wall tapestry weight, filters, and extracting solution reclaims ethanol, be concentrated into relative density d=1.05, filter, obtain medical liquid B;
(3) by medical liquid B that step (2) obtains, by D101 nonpolarity macroporous adsorptive resins chromatographic column, first wash with water, water elution liquid is directly by the polar macroporous resin column of DM130, use the alcoholic solution eluting D101 nonpolarity macroporous adsorptive resins chromatographic column of concentration 60% again, collect 60% concentration ethanol eluent, concentrate drying, obtain wall tapestry extract B; Use the alcoholic solution eluting DM130 polar macroporous adsorbent resin column of concentration 85% again, collect 85% concentration ethanol eluent, concentrate drying, obtain wall tapestry extract C;
(4) by above-mentioned wall tapestry extract A, wall tapestry extract B, the mixing of wall tapestry extract C, wall tapestry extract of the present invention is obtained.
Embodiment 4: the preparation of wall tapestry extract sheet
Example 1 wall tapestry extract 245g, adds starch 80g, mixing, granulates, dry, sieves, adds microcrystalline Cellulose 15g, magnesium stearate 2.5g, and mixing, is pressed into 1000, obtains wall tapestry extract sheet.
Embodiment 5: the preparation of wall tapestry extract particles
Example 2 wall tapestry extract 235g, adds dextrin 95g, mixing, granulates, and dry, granulate, obtains wall tapestry extract particles.
Embodiment 6: the preparation of wall tapestry extract capsule
Example 3 wall tapestry extract 255g, adds starch 35g, galactose granulation 25g, mixing, granulates, and dry, granulate, obtains wall tapestry extract capsule by encapsulated 1000.
Experimental example 1: wall tapestry extract is to acquired immune deficiency syndrome (AIDS) mice therapeutical effect.
1. materials and methods
Medicine: wall tapestry extract, wall tapestry extract A, wall tapestry extract B, wall tapestry extract C, be embodiment 1 method to prepare, lot number is respectively: 040810,040811,040812,040813, dissolve with the dimethyl sulfoxide of 30g/L (DMSO) during test, then with distilled water diluting to desired concn; Positive control medicine zidovudine (AZT) is produced by Xiamen Pharmaceutical Factory, and lot number is 030907.
Animal: only, female, weight l3-15g, is provided by Guangdong Province's Experimental Animal Center, the quality certification number SPF level BALB/C mice 5O: check and affirm word 2005A013 in SCXK (Guangdong) 2003-0002 Guangdong.
Seed culture of viruses: Friend type murine leukemia virus (FriendLeukemia virus, FLV) draw from American Type culture collecting center (American Typical Culture Centre, ATCC), after mice goes down to posterity enhancing virulence, make 100 g/L spleen suspensions ,-70 DEG C frozen for subsequent use.
Main agents: the two labeling antibody of PE/Cy5 anti-mouse CD3e monoclonal; PE/Cy5 Mouse IgG1, K Isotype Ctrl Isotype control; FITC anti-mouse CD4/PE anti-mouse CD8a monoclonal list labeling antibody; FITC Mouse IgG1, K Isotype Ctrl/PEMouse IgG1, K Isotype Ctrl Isotype control, is more than U.S. Biolegend Products.
Virus median infective dose (ID
50) titration: lumbar injection FLV, makes 0.1g/L spleen suspension, collects spleen suspension and press literature method calculating ID
50, with 60 times of ID during experiment
50for the experimental concentration of mouse model of AIDS.
The each extract of wall tapestry causes the inhibitory action of mice spleen enlargement to FLV: 70 mices are divided into normal group, model group, wall tapestry extract group, wall tapestry extract A group, wall tapestry extract B group, wall tapestry extract C group, AZT group, often organize each 10; Except normal group, all adopt FLV lumbar injection infecting mouse.Normal group and model group every day are with distilled water 0.2 mL/ gavage, wall tapestry extract group, wall tapestry extract A group, wall tapestry extract B group, wall tapestry extract C group dosage are 20mg/kg, AZT group dosage is 100mg/kg, all be made into 2mL solution with distilled water, after virus attack, l d starts with the administration of gavage mode, 0.2 mL/, 1 time/d, successive administration 21d; Cut open after administration 21d and kill mice, calculate mouse spleen index and spleen index suppression ratio, spleen index (mass ratio) ω
spleen=m
spleen(mg)/m
body(g), p
spleen index suppresses=(ω
on average, model group-ω
on average, administration group)/ω
on average, model group× 100%.Obtained experimental data statistic software SPSS 10.0 is analyzed, compares between each group and adopt one factor analysis of variance (LSD).
Hemogram checking: pluck eyeball anticoagulant and get blood, carries out erythrocyte (RBC) counting, leukocyte (WBC) counting with full-automatic blood counting instrument.
Impact on cellular immune function: measure mouse thymus index (mass ratio), ω
thymus=m
thymus(mg)/m
body(g); T lymphocyte subsets of peripheral blood horizontal detection, adopts cells were tested by flow cytometry BALB/C mice T lymphocyte subsets CD3
+, CD4
+, CD8
+, CD4
+/ CD8
+level.
2. result.
2.1 each groups cause the inhibiting comparison of mice spleen enlargement to FLV
The spleen index of each extract group of wall tapestry has significant difference (P<0.05) compared with model group, it is inhibited that display wall tapestry extract etc. causes mice spleen enlargement to FLV, and suppression ratio is respectively: 94.45%, 85.33%, 94.18%, 81.98%.The results are shown in Table 1.
The each extract of table 1 wall tapestry is to the inhibitory action of FLV infecting mouse splenomegaly
1.: P<0.05,2.: P<0.0l, compare with Normal group; 3.: P<0.05,4.: P<0.0l, compare with model group (lower same).
The comparison of 2.2 each group cellular immune functions
Wall tapestry each extract group thymus is better protected.Do not occur obvious atrophy, the decline of the thymus index that can cause after antagonism viral infection, shows good immanoprotection action.Periphery blood T lymphocyte analyzes (TLC) display, and wall tapestry extract etc. can significantly improve peripheral blood CD3
+, CD4
+, CD8
+t lymphocyte level.The results are shown in Table 2.
The each extract of table 2 wall tapestry is on the impact of the cellular immune function of FLV infecting mouse.
This experimental result shows, the splenomegaly that wall tapestry extract oral can suppress FLV to cause.Spleen is the first target organs of virus, and the spleen of FLV infected mice increases weight large I quantitative response virus quantity, and therefore can medicine suppress virus, and spleen index is a directly perceived and index reliably.Spleen has abundant lymphocyte and macrophage, and they all participate in humoral immune reaction, so wall tapestry extract can suppress splenomegaly in an experiment, shows it and not only has anti-FLV effect in body, also may have the effect improving humoral immune function.From the experimental result of amynologic index, wall tapestry extract has good rise effect to cellular immunization, particularly when the body's immunity that retrovirus causes is low, performance is particularly remarkable, showing this medicine can enhancing body cellular immune function, suppressing the immunodeficiency that FLV causes, is a good immunomodulator.
Claims (8)
1. the application of wall tapestry extract in preparation treatment AIDS-treating medicine, the preparation method of described wall tapestry extract is:
(1) wall tapestry, with concentration 80%-100% ethanol as solvent, extract at 20 DEG C of-70 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 wall tapestry weight, filters, obtains medicinal residues A and extracting solution, extracting solution reclaims ethanol, concentrated, dry, obtain wall tapestry extract A;
(2) step (1) being obtained medicinal residues A concentration 0-50% ethanol is solvent, extract at 30 DEG C of-80 DEG C of warm macerating, extraction time is 2-5 time, each extraction time is 1-6 hour, and each solvent load is 6-20 times of wall tapestry weight, filters, extracting solution reclaims ethanol, be concentrated into relative density d=1.05-1.12, filter, obtain medical liquid B;
(3) by medical liquid B that step (2) obtains, pass through nonpolarity macroporous adsorptive resins chromatographic column, first wash with water, water elution liquid is directly by polar macroporous resin column, use the alcoholic solution eluting nonpolarity macroporous adsorptive resins chromatographic column of concentration 60%-95% again, collect different concentration ethanol eluent, concentrate drying, obtains wall tapestry extract B; Use the alcoholic solution eluting polar macroporous adsorbent resin column of concentration 60%-85% again, collect different concentration ethanol eluent, concentrate drying, obtains wall tapestry extract C;
(4) above-mentioned wall tapestry extract A, wall tapestry extract B, wall tapestry extract C, wherein one or both or three kinds mix by a certain percentage, obtain wall tapestry extract.
2. application according to claim 1, is characterized in that the preparation method of described wall tapestry extract is:
(1) wall tapestry, with concentration 80%-100% ethanol as solvent, extract at 20 DEG C of-70 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 wall tapestry weight, filters, obtains medicinal residues A and extracting solution, extracting solution reclaims ethanol, concentrated, dry, obtain wall tapestry extract A;
(2) step (1) being obtained medicinal residues A concentration 0-50% ethanol is solvent, extract at 30 DEG C of-80 DEG C of warm macerating, extraction time is 2-5 time, each extraction time is 1-6 hour, and each solvent load is 6-20 times of wall tapestry weight, filters, extracting solution reclaims ethanol, be concentrated into relative density d=1.05-1.12, filter, obtain medical liquid B;
(3) by medical liquid B that step (2) obtains, pass through nonpolarity macroporous adsorptive resins chromatographic column, first wash with water, water elution liquid is directly by polar macroporous resin column, use the alcoholic solution eluting nonpolarity macroporous adsorptive resins chromatographic column of concentration 60%-95% again, collect different concentration ethanol eluent, concentrate drying, obtains wall tapestry extract B; Use the alcoholic solution eluting polar macroporous adsorbent resin column of concentration 60%-85% again, collect different concentration ethanol eluent, concentrate drying, obtains wall tapestry extract C;
(4) by above-mentioned wall tapestry extract A, wall tapestry extract B, the mixing of wall tapestry extract C, wall tapestry extract is obtained.
3. application according to claim 1, is characterized in that the preparation method of described wall tapestry extract is:
(1) wall tapestry, with concentration 90% ethanol as solvent, extract at 50 DEG C of warm macerating, extraction time is 4 times, each extraction time is 4 hours, and each solvent load is 20 times of wall tapestry weight, filters, obtains medicinal residues A and extracting solution, extracting solution reclaims ethanol, concentrated, dry, obtains wall tapestry extract A;
(2) step (1) being obtained medicinal residues A concentration 30% ethanol is solvent, and extract at 70 DEG C of warm macerating, extraction time is 3 times, and each extraction time is 3 hours, each solvent load is 15 times of wall tapestry weight, filters, and extracting solution reclaims ethanol, be concentrated into relative density d=1.12, filter, obtain medical liquid B;
(3) by medical liquid B that step (2) obtains, by AB-8 nonpolarity macroporous adsorptive resins chromatographic column, first wash with water, water elution liquid is directly by the polar macroporous resin column of DM130, use the alcoholic solution eluting AB-8 nonpolarity macroporous adsorptive resins chromatographic column of concentration 85% again, collect 85% concentration ethanol eluent, concentrate drying, obtain wall tapestry extract B; Use the alcoholic solution eluting DM130 polar macroporous adsorbent resin column of concentration 70% again, collect 70% concentration ethanol eluent, concentrate drying, obtain wall tapestry extract C;
(4) by above-mentioned wall tapestry extract A, wall tapestry extract B, the mixing of wall tapestry extract C, wall tapestry extract is obtained.
4. application according to claim 1, is characterized in that adopted nonpolar macroporous adsorption resin is AB-8 macroporous adsorbent resin, D101 macroporous adsorbent resin; The polar macroporous adsorption resin adopted is DM130 macroporous adsorbent resin, LSA-10 macroporous adsorbent resin.
5. application according to claim 1, is characterized in that the treatment AIDS-treating medicine that described wall tapestry extract and chemical drugs or Chinese medicine or natural drug form.
6. application according to claim 1, is characterized in that the application in preparation treatment AIDS-treating medicine of described wall tapestry extract A, wall tapestry extract B, wall tapestry extract C.
7. application according to claim 1, is characterized in that the treatment AIDS-treating medicine that described wall tapestry extract A, wall tapestry extract B, wall tapestry extract C and chemical drugs or Chinese medicine or natural drug form.
8. application according to claim 1, is characterized in that described wall tapestry extract makes tablet oral on pharmaceutics, granule, capsule by the various adjuvants adding pharmaceutics permission.
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| "Lichen-derived Culture and Its Pharmaceutical Applications";Yoshikazu Yamamoto;《International Symposium on Systematic Lichenology Bryology》;20101021;82-85 * |
| 张璞 等."地衣抗生素和地衣多糖类物质的研究进展".《山东科学》.2007,第20卷(第2期), * |
| 房敏峰等."地衣化学成分和药理作用研究进展".《中草药》.2011,第42卷(第12期), * |
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