CN102727654A - Pharmaceutical composition providing inhibition effect against inflammation factor expression - Google Patents
Pharmaceutical composition providing inhibition effect against inflammation factor expression Download PDFInfo
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Abstract
The invention provides a pharmaceutical composition providing an inhibition effect against inflammation factor expression. The pharmaceutical composition is composed of a first extract, a second extract and a third extract. The preparation method of the extracts comprises the steps that: raw materials of honeysuckle flower, figwort, Chinese angelica and licorice are well mixed according to a weight ratio of 2.5-4:2.5-4:1.5-2.5:0.5-1.5; the mixture is subjected to ethanol extraction, such that an extraction liquid is obtained; the extraction liquid is condensed, and is loaded on macroporous resin; 30% ethanol, 60% ethanol, and 95% ethanol are respectively used for eluting, such that the first extract, the second extract and the third extract are obtained; the extracts are mixed, such that the pharmaceutical composition is obtained. The pharmaceutical composition provided by the invention can effectively inhibit inflammation factor expression.
Description
Technical field
The present invention relates to field of medicaments, especially effective ingredient in Chinese and application thereof especially relate to a kind of compositions of inflammatory factor being expressed inhibited effective ingredient in Chinese.
Background technology
Inflammation damnification is internal organs, when damage takes place, by the complex reaction of immune system wide participation, can discharge a large amount of inflammatory factors in the inflammatory process like heart, brain etc., comprises cytokine, adhesion molecule etc., impels the further generation of damage.The generation of inflammatory reaction can cause the generation of internal organs local secondary damage, and for example cerebral ischemia, myocardial ischemia all are the common clinicals that have close ties with inflammatory reaction.
For example, acute coronary syndrome (ACS) is one group of clinical syndrome that is caused by acute myocardial ischemia, and coagulation function activates and the vulnerable plaque inflammatory reaction plays an important role in the generation of ACS, is two core pathology links.And for example in the apoplexy process, tangible inflammatory reaction all appears in cerebral ischemia and again after the perfusion in the cerebral tissue, causes the secondary damage after the cerebral ischemia.
Chinese medicine cardiac-cerebral ischemia sexually transmitted disease (STD) has shown great potential on becoming, and still, ideal medicine does not also develop." SIMIAOYONGAN TANG " is made up of " Flos Lonicerae, Radix Scrophulariae, Radix Angelicae Sinensis, Radix Glycyrrhizae ", has the invigorate blood circulation effect of analgesic therapy of remarkable detoxifcation, and tradition is used for artery of lower extremity obliterans and vasculitic treatment.In recent years, doctor family is based on to internal organs, the for example heart, brain, and the understanding of ischemic inflammatory reaction pathomechanism begins this side to be used for treatments such as coronary heart disease, cerebral ischemia, has obtained satisfied clinical efficacy." SIMIAOYONGAN TANG " compatibility is precise and appropriate, and medicine is concentrated one's efforts grand, is one of tradition name side of tool traditional Chinese medical science compatibility and consumption characteristic; Has good patent medicine DEVELOPMENT PROSPECT; Its compatibility rule and dose-effect relationship also are worth further investigation, but because former side's dosage is excessive, bring difficulty to new drug development.
Therefore; Need to find the pharmacodynamics material base of tradition name side; Find its active site place, develop that effective substance is clear, the effect link is clear and definite, safe and efficient, quality controllable, compatibility is precise and appropriate to the inhibited modern medicines compositions of inflammatory factor.
Summary of the invention
In order to reach above-mentioned at least one goal of the invention, the invention provides and a kind of inflammatory factor is expressed inhibited pharmaceutical composition, the ingredient of said composition is the effective site of traditional Chinese medical science tradition name side " SIMIAOYONGAN TANG ".
Provided by the present invention to the inhibited pharmaceutical composition of inflammatory factor expression; Be made up of the 1st extract, the 2nd extract and the 3rd extract, said preparation method of extract is following: a) with crude drug Flos Lonicerae, Radix Scrophulariae, Radix Angelicae Sinensis, Radix Glycyrrhizae according to weight ratio 2.5～4: 2.5～4: 1.5～2.5: 0.5～1.5 ratio mixing; B) with the ethanol that adds entry or 40～60% in the said crude drug as extracting solvent, the solid-to-liquid ratio of said crude drug and said extraction solvent is 1: 5～10, extract more than 2 times, each 1.5～4.5 hours, filter merging filtrate afterwards, obtain extracting solution and residue; C) be concentrated into said extracting solution dried; After add the water suspendible; Last macroporous resin; Adopt water, 30% ethanol, 60% ethanol and 95% ethanol to carry out continuous drip washing as eluent, obtain 30% ethanol drip washing thing respectively and be said the 1st extract, 60% ethanol drip washing thing is said the 2nd extract, 95% ethanol drip washing thing is said the 3rd extract; D) said the 1st, 2,3 extracts are mixed, obtain said pharmaceutical composition.
One of according to the embodiment of the present invention, the preferred weight ratio of Flos Lonicerae, Radix Scrophulariae, Radix Angelicae Sinensis, Radix Glycyrrhizae is 3: 3: 2 in the crude drug: 1.This is the summary of clinical experience, is again simultaneously the effective proportioning through experimental verification.
According to another embodiment of the present invention, concentration of ethanol is 55% in the step b) of extraction.
One of according to the embodiment of the present invention, in the step b) solid-to-liquid ratio for be preferably 1: 6～8.
One of according to the embodiment of the present invention, the extraction time in the step b) is 4 times, each extraction time is 2～4 hours.
One of according to the embodiment of the present invention, the macroporous resin that is adopted in this method for preparing is the AB-8 macroporous resin.
Preferably; Provided by the present invention a kind of to the inhibited pharmaceutical composition of inflammatory factor expression; Be made up of the 1st extract, the 2nd extract and the 3rd extract, said preparation method of extract is following: a) with crude drug Flos Lonicerae, Radix Scrophulariae, Radix Angelicae Sinensis, Radix Glycyrrhizae according to weight ratio 3: 3: 2: 1 ratio mixing; B) with adding 55% ethanol in the said crude drug as extracting solvent, the solid-to-liquid ratio of said crude drug and said extraction solvent is 1: 8, extracts 4 times, and each 2 hours, filtration is merging filtrate afterwards, obtains extracting solution and residue; C) be concentrated into said extracting solution dried; After add the water suspendible; Last AB-8 macroporous resin; Adopt water, 30% ethanol, 60% ethanol and 95% ethanol to carry out continuous drip washing as eluent, obtain 30% ethanol drip washing thing and be said the 1st extract, 60% ethanol drip washing thing is said the 2nd extract, 95% ethanol drip washing thing is said the 3rd extract; D) said the 1st～3 extract is mixed, obtain said pharmaceutical composition.
The above pharmaceutical composition provided by the present invention and preferred version thereof are that the inventor gropes to form on the basis of medical practice for many years, and through improvement dosage form experimental verification, that be directed against tradition name side SIMIAOYONGAN TANG.
Extract 1～3 provided by the present invention is expressed all inhibited through experimental verification to inflammatory factor.Through the inventor in experiment; Adopt NF-κ B reaction original paper binding inhibitors screening model and peroxidase multiplication agent receptor (PPAR γ) agonist screening model; Effect to the reaction of the inflammation-inhibiting of these extracts is verified, confirms that they all have the effect of inhibition atherosclerotic plaque inflammatory reaction in various degree.
NF-κ B overactivity can cause multiple pathological reaction, and is particularly relevant with various inflammatory reactions, and wherein activating the atherosclerotic plaque inflammatory reaction also is one of them.NF-κ B is many to combine the form of p65 subunit to exist with p50 or p52, and p65 subunit and I κ B dissociate after the activation, get into karyon by the endochylema transposition, combines the expression of adjusting genes of interest with reaction original paper (RE) on the target gene.Therefore, the blocking strategy that early gene is expressed after the NF-κ B transposition is for suppressing the crucial target spot of its pathological signals path.The inventor utilizes molecule clone technology; With Human umbilical vein endothelial cells cDNA is template, and amplification NF-κ B-RE gene is cloned in the pGL4.32 carrier for expression of eukaryon; Through transfection; Import in the 293A cell, screen, go down to posterity, set up the cell line (pGL4.32 [luc2P/NF-kappaB-RE] 293A) of stably express NF-κ B-RE through Hygromycin.This model can directly detect the variation of luciferase photochemistry value through the luciferase detectable, be used to carry out the high flux screening of NF-kB inhibitor.Empirical tests confirms that the 1st, 2 extracts have blocking effect to NF-κ B path, can suppress this pathological signals path, IC through suppressing the early gene expression
50Be respectively 31.61 μ g/ml and 32.49 μ g/ml.
Peroxidase multiplication agent receptor (PPAR) is one of the activated transcription factor nuclear receptor superfamily of part member.Present known its three hypotypes: PPAR α ,-β/δ and-γ.Peroxidase multiplication agent receptor y (PPAR γ) can influence the signal path of NF-κ B, signal transcription, activator protein-1 mediation, reaches through the activation that suppresses these approach and suppresses the purpose that target gene promoters activates and transcribes.An ability is contained by the activated protein kinase phosphorylation of mitogen site in the N of PPAR γ end functional areas, if this zone sudden change or can not produce phosphorylation and make its activation in that the phosphoprotein phosphatase combined effect is next.After part combines with PPAR γ and makes it to activate; With retinal X receptor α (retinoid Xreceptor α; RXR α) forms heterodimer; Be incorporated into specific DNA sequences again and make target gene activation, this sequence be called PPAR specific reaction element (peroxisome proliferator responsiveelement, PPRE).The gene that contains the PPRE structure comprises acyl-CoA synthetase, lipoprotein lipase (LPL), IRS-2 (IRS-2), leptin and tumor necrosis factor-alpha (TNF-α) etc.PPAR γ, at lipogenesis, glycolipid metabolism and plays a significant role in immune system through regulating Expression of Related Genes, and relevant with generation, the development of multiple disease such as diabetes, obesity, hypertension, cancer etc.Especially PPAR γ is the key factor in the adipose cell atomization, receives much concern in recent years.PPAR γ becomes obesity, hypertension, atherosclerosis therapy target spot.The inventor is its agonist of target position screening with PPAR γ, finds that the 1-3 extract all has PPAR gamma agonist effect in various degree.The most tangible with the 3rd extract effect, its effective percentage when 0.01mg/ml is 90%, EC
500.004mg/ml; The effective percentage of the 2nd extract when 0.1mg/ml is 100%, EC
500.01mg/ml; The effective percentage less than 60% of the 1st extract.
According to experimental result; It is thus clear that the 1st～3 extract that is contained in the pharmaceutical composition provided by the present invention all has the activatory peroxisome proliferation-activated receptors γ of promotion (PPAR-γ) in various degree to express and/or inhibition nuclear factor κ B (NF-κ B) expression.Wherein, the most remarkable with the effect that promotes PPAR-γ to express.Thereby; Medication needs according to clinical practice; With these extracts carry out selectively, the combination of arbitrary proportion, can obtain promptly that effective substance is clear, the effect link is clear and definite, safe and efficient, quality controllable, compatibility is precise and appropriate to the inhibited modern medicines compositions of inflammatory factor.
In order further to improve the effect of the inflammation-inhibiting factor of this pharmaceutical composition, the inventor has carried out research further to the compatibility rule of three kinds of effective sites, has found the proportion compatibility that some are better.
One of according to the embodiment of the present invention, in the aforementioned pharmaceutical compositions, said the 1st extract, the 2nd extract, the 3rd extract are according to weight ratio 0.3～5.4: 0.2～4.2: 1 ratio is mixed.
According to another embodiment of the present invention, in the aforementioned pharmaceutical compositions, said the 1st extract, the 2nd extract, the 3rd extract are according to weight ratio 0.6～3: 0.5～2.1: 1 ratio is mixed.
According to an embodiment more of the present invention, in the aforementioned pharmaceutical compositions, said the 1st extract, the 2nd extract, the 3rd extract mix according to 1.34: 1.05: 1 ratio of weight ratio.
The inventor is through the different orthogonal test checkings with concentration of 3 kinds of extracts, and according to the pharmaceutical composition that the aforementioned proportion range fit forms, performance is stablized aspect the inflammation-inhibiting factor, and effect is more outstanding.
Additional aspect of the present invention and advantage part in the following description provide, and part will become obviously from the following description, or recognize through practice of the present invention.
The specific embodiment
Describe embodiments of the invention below in detail.The embodiment that describes below is exemplary, only is used to explain the present invention, and can not be interpreted as limitation of the present invention.
Experimental example one single extract is to the combination inhibitory action of NF-κ B reaction original paper
1, experimental procedure
(1) experiment material: NF-κ B-RE cell (pGL4.32 [luc2P/NF-kappaB-RE] 293A; Institute of Materia Medica,Chinese Academy of Medical Sciences), MEM-α culture medium (Gibco), hyclone (Gibco); 96 well culture plate (Costar; 3599), Bright-Glo Luciferase luciferase detectable (Promaga), water-jacket typ CO
2Cell culture incubator (Sanyo, Japan), Safire2 high-speed multiple channel continuous wavelength ELIASA (TECAN, Austria).
(2) test sample:
Adopt embodiment 1 said method extract obtained 1～3, provide by Beijing University of Chinese Medicine.
(3) experimental procedure:
With the NF-κ B-RE cell dissociation and the counting of logarithmic growth in the culture bottle, with 100 μ l systems, 5 * 10
3The density of/well is inoculated in the white culture plate of 96 holes, hatches 18-24h.Matched group changes the culture medium with serum-free MEM-α; Wait to sieve sample and behind 18h, add simultaneously, hatch 12h jointly with cell then with the concentration of 100 μ g/ml, 50 μ g/ml, 25 μ g/ml, 10 μ g/ml, 5 μ g/ml.After sample effect finishes, place 30min in room temperature, inhale and abandon former culture medium, every hole adds 50 μ l Bright-GloLuciferase luciferase detectable, and room temperature lucifuge concussion 5min places the photochemistry detector, selects the ONE-Glo program to detect.
This model directly indicates sample whether NF-κ B-RE is had blocking effect with the variation of luciferase photochemistry value.Therefore, through calculating suppression ratio, can confirm sample power to early gene expression blocking effect in NF-κ B signal path.
2, experimental result
Detailed results is seen shown in the table 1.
Table 1 extract is to the effect of NF-κ B reaction original paper binding inhibitors screening model
(data represent with mean ± SEM, n=3)
According to table 1, visible extract 1 and 2 extracts have blocking effect to NF-κ B signal path, can suppress this pathological signals path, its IC through suppressing the early gene expression
50Be respectively 31.61 μ g/ml, 32.49 μ g/ml.
Experimental example two single extracts are to the agonism of peroxidase multiplication agent receptor (PPAR γ)
1 material
1.1 reagent:
Africa green ape and monkey nephrocyte (CV-1), PPRE plasmid, PPAR plasmid, RXR α plasmid, MEM-α culture medium (Gibco), hyclone (Gibco company), Bright-Glo Luciferase luciferase detectable (Promega company), rosiglitazone.
1.2 instrument:
Water-jacket typ CO2 cell culture incubator (Sanyo, Japan), EnSpireTM 2300MultilabelReader (PerkinElmer company).
2 methods
2.1 the extraction of test sample
Adopt extract obtained 3 of embodiment 1 said method, provide by Beijing University of Chinese Medicine.
2.2 cell culture
The green ape and monkey nephrocyte in Africa (CV-1) uses the DMEM culture medium that is added with 10%FBS and 1% antibiotic (penicillin/streptomycin) at 5%CO
2Cultivate in 37 ℃ of the incubators.
2.3 transfection and luciferase gene expression horizontal detection
PPRE luciferase reporter gene plasmid and PPAR γ and the RXR α plasmid co-transfection that will contain PPAR target sequence response element are gone in the African green ape and monkey nephrocyte (CV-1); After the transfection 24 hours; With the 30000cells/well bed board; Add positive drug and testing compound after 4 hours, (Rosiglitazone) is the agonist positive control with rosiglitazone, in CO2 gas incubator, cultivates and detects the luciferase gene expression level after 24 hours.
3 experimental results
The result gathers and sees table 2, and it is as shown in the table: with the inductive reporter gene activity of 20 μ M rosiglitazones (Rosiglitazone) is 100%; Relative activity % and the EC of chemical compound when 1mg/ml
50(mg/ml):
Table 2
The result shows:
In the exciting model, as initial concentration, the 1st extract is not seen cell toxicant with 1mg/ml, Efficacy% (relative activity %)＜60%; The 2nd extract is seen obvious cell toxicant at 1mg/ml, and the Efficacy% during 0.1mg/ml is 100%; The 3rd extract is 1, and 0.1mg/ml all sees obvious cell toxicant, and the Efficacy% during 0.01mg/ml is 90%.
4 discuss:
PPAR γ can suppress the expression of atherosclerosis (AS) speckle proinflammatory factor through many A signal pathways, alleviates immunity and inflammatory reaction, suppresses monocyte/macrophage to the foam cell conversion etc., thereby delays the AS development, and stabilize plaque prevents to break.PPAR γ has become the novel targets of anti-AS new drug development.PPAR γ is the key link of inflammation-inhibiting reaction.
Experimentation finds that the inductive reporter gene activity of positive control medicine 20 μ M rosiglitazones (Rosiglitazone) is 100%, has tangible activation.In the exciting model that research is set up, the 2nd extract provided by the invention and the 3rd extract are remarkably productive.
Experimental example three extracts are to the combination inhibitory action compatibility rule research of NF-κ B reaction original paper
1. experimental procedure
With experimental example one.
2. orthogonal design
3 factors, 3 horizontal quadrature EXPERIMENTAL DESIGN are adopted in the research of compatibility rule, according to orthogonal table, design 27 compatibility groups altogether.Wherein 3 factors are the 1st extract, the 2nd extract, the 3rd extract, and 3 levels are got concentration point, factor level table such as table 3 in each active site live vol effect relationship scope.
Table 3
| (μg/ml) | The 1st extract | The 2nd extract | The 3rd extract |
| 1 | 69 | 54 | 51.6 |
| 2 | 34.5 | 27 | 25.8 |
| 3 | 17.25 | 13.5 | 12.9 |
3. experimental result
Detailed results is seen table 4.
Each compatibility group of table 4 is to the effect of NF-κ B reaction original paper binding inhibitors screening model
Data are represented with mean ± SEM, n=4.
According to table 4; It is thus clear that 2,3,6,8,9,10,13,15,16,17,18,23,24,25,26, No. 27 compatibility groups of compatibility group can improve suppression ratio; Explain that above compatibility has blocking effect to NF-κ B signal path, can suppress this pathological signals path through suppressing the early gene expression.Wherein 8,9,18, No. 26 compatibility groups of compatibility group can obviously improve suppression ratio, and prompting has strong blocking effect to NF-κ B signal path.And behind 1,4, No. 7 compatibility group effects of compatibility group 12h NF-κ B-RE is shown tangible toxic action, though above-mentioned 3 kinds of compatibilities can improve suppression ratio, not the blocking effect of medicine itself.
Experimental example four extracts are to the agonism compatibility rule research of peroxidase multiplication agent receptor (PPAR γ)
1. experimental procedure
(1) experiment material:
With experimental example two.
(2) test sample:
Adopt embodiment 1 said method extract obtained 1～3, provide by Beijing University of Chinese Medicine.
Adopt 3 factors, 3 horizontal quadrature EXPERIMENTAL DESIGN, design is with experimental example three.
(3) experimental procedure:
With experimental example two.
2, experimental result
Detailed results is seen shown in the table 5.
Experimental result is following: with the inductive reporter gene activity of 20 μ M Rosiglitazone is 100%.
Table 5 compatibility group is to the effect of the agonist screening model of PPAR γ
*: twice experiment (respectively) average data;
Efficacy%: relative activity %.
The result shows; 1,2,3,5,7,8,10,11,12,13,14,16,17,19,20,22,23,25, No. 26 compatibility groups all have PPAR gamma agonist effect in various degree; Explain that above compatibility has activation to PPAR γ signal path, thereby the early gene that suppresses NF-κ B reaches this pathological signals path of inhibition.
Wherein 1,2,4,5,7,10,12,13,16,19,20,22, No. 25 compatibility group has tangible activation, and prompting has strong activation to PPAR γ signal path.
Embodiment one
With crude drug Flos Lonicerae 3.6g, Radix Scrophulariae 3.6g, Radix Angelicae Sinensis 2.4g, Radix Glycyrrhizae 1.2g, pulverize the back mix homogeneously, add the ethanol of 8 times of amounts 55%, extract each 2 hours 4 times.Merging filtrate after extraction finishes obtains 55% ethanol extract and residue.
Be concentrated into 55% ethanol extract dried; After add the water suspendible; Last AB-8 macroporous resin; Adopt water, 30% ethanol, 60% ethanol and 95% ethanol to carry out continuous drip washing as eluent, obtain 30% ethanol drip washing thing respectively and be the 1st extract, 60% ethanol drip washing thing is the 2nd extract, 95% ethanol drip washing thing is the 3rd extract.
The 1st extract, the 2nd extract and the 3rd extract are mixed according to 345: 270: 258 ratio, promptly get.
Embodiment two
With crude drug Flos Lonicerae 7.5g, Radix Scrophulariae 7.5g, Radix Angelicae Sinensis 4.5g, Radix Glycyrrhizae 1.5g, pulverize the back mix homogeneously, add 40% ethanol of 5 times of amounts, extract each 1.5 hours 3 times.Merging filtrate after extraction finishes obtains 40% ethanol extract and residue.
Be concentrated into 40% ethanol extract dried; After add the water suspendible; Last AB-8 macroporous resin; Adopt 30% ethanol, 60% ethanol and 95% ethanol to carry out continuous drip washing as eluent, obtain 30% ethanol drip washing thing respectively and be the 1st extract, 60% ethanol drip washing thing is the 2nd extract, 95% ethanol drip washing thing is the 3rd extract.
The 1st extract, the 2nd extract and the 3rd extract are mixed according to 1.3: 1.1: 1 ratio, promptly get.
Embodiment three
With crude drug Flos Lonicerae 12g, Radix Scrophulariae 12g, Radix Angelicae Sinensis 7.5g, Radix Glycyrrhizae 4.5g, pulverize the back mix homogeneously, add 60% ethanol of 10 times of amounts, extract each 4.5 hours 2 times.Merging filtrate after extraction finishes obtains 60% ethanol extract and residue.
Be concentrated into 60% ethanol extract dried; After add the water suspendible; Last AB-8 macroporous resin; Adopt water, 30% ethanol, 60% ethanol and 95% ethanol to carry out continuous drip washing as eluent respectively, obtain 30% ethanol drip washing thing respectively and be the 1st extract, 60% ethanol drip washing thing is the 2nd extract, 95% ethanol drip washing thing is the 3rd extract.
The 1st extract, the 2nd extract and the 3rd extract are mixed according to 2.7: 2.1: 1 ratio, promptly get.
Embodiment four
With crude drug Flos Lonicerae 11.4g, Radix Scrophulariae 8.4g, Radix Angelicae Sinensis 5.7g, Radix Glycyrrhizae 3.9g, pulverize the back mix homogeneously, add 58% ethanol of 7 times of amounts, extract each 2 hours 4 times.Merging filtrate after extraction finishes obtains 58% ethanol extract and residue.
Be concentrated into 58% ethanol extract dried; After add the water suspendible; Last AB-8 macroporous resin; Adopt 30% ethanol, 60% ethanol and 95% ethanol to carry out continuous drip washing as eluent, obtain 30% ethanol drip washing thing respectively and be the 1st extract, 60% ethanol drip washing thing is the 2nd extract, 95% ethanol drip washing thing is the 3rd extract.
The 1st extract, the 2nd extract and the 3rd extract are mixed according to 5.4: 4.2: 1 ratio, promptly get.
Embodiment five
With crude drug Flos Lonicerae 8.1g, Radix Scrophulariae 10.8g, Radix Angelicae Sinensis 6.9g, Radix Glycyrrhizae 2.4g, pulverize the back mix homogeneously, add 50% ethanol of 6 times of amounts, extract each 2.5 hours 5 times.Merging filtrate after extraction finishes obtains 50% ethanol extract and residue.
Be concentrated into 50% ethanol extract dried; After add the water suspendible; Last AB-8 macroporous resin; Adopt water, 30% ethanol, 60% ethanol and 95% ethanol to carry out continuous drip washing as eluent respectively, obtain 30% ethanol drip washing thing respectively and be the 1st extract, 60% ethanol drip washing thing is the 2nd extract, 95% ethanol drip washing thing is the 3rd extract.
The 1st extract, the 2nd extract and the 3rd extract are mixed according to 0.6: 2.1: 1 ratio, promptly get.
Embodiment six
With crude drug Flos Lonicerae 9.9g, Radix Scrophulariae 9.9g, Radix Angelicae Sinensis 6g, Radix Glycyrrhizae 3g, pulverize the back mix homogeneously, add 45% ethanol of 8 times of amounts, extract each 4 hours 4 times.Merging filtrate after extraction finishes obtains 45% ethanol extract and residue.
Be concentrated into 45% ethanol extract dried; After add the water suspendible; Last AB-8 macroporous resin; Adopt 30% ethanol, 60% ethanol and 95% ethanol to carry out continuous drip washing as eluent, obtain 30% ethanol drip washing thing respectively and be the 1st extract, 60% ethanol drip washing thing is the 2nd extract, 95% ethanol drip washing thing is the 3rd extract.
The 1st extract, the 2nd extract and the 3rd extract are mixed according to 3: 0.5: 1 ratio, promptly get.
Embodiment seven
The 1st extract, the 2nd extract and the 3rd preparation method of extract are with embodiment 1.
After obtaining 3 kinds of extracts, the 1st extract, the 2nd extract and the 3rd extract are mixed according to 5.4: 1.1: 1 ratio, promptly get.
Embodiment seven
The 1st extract, the 2nd extract and the 3rd preparation method of extract are with embodiment 2.
After obtaining 3 kinds of extracts, the 1st extract, the 2nd extract and the 3rd extract are mixed according to 0.7: 0.5: 1 ratio, promptly get.
Embodiment eight
The 1st extract, the 2nd extract and the 3rd preparation method of extract are with embodiment 3.
After obtaining 3 kinds of extracts, the 1st extract, the 2nd extract and the 3rd extract are mixed according to 0.7: 1: 1 ratio, promptly get.
Although illustrated and described embodiments of the invention; For those of ordinary skill in the art; Be appreciated that under the situation that does not break away from principle of the present invention and spirit and can carry out multiple variation, modification, replacement and modification to these embodiment, scope of the present invention is limited accompanying claims and equivalent thereof.
Claims (9)
1. one kind inflammatory factor expressed inhibited pharmaceutical composition, be made up of the 1st extract, the 2nd extract and the 3rd extract, said preparation method of extract is following:
A) with crude drug Flos Lonicerae, Radix Scrophulariae, Radix Angelicae Sinensis, Radix Glycyrrhizae according to weight ratio 2.5～4: 2.5～4: 1.5～2.5: 0.5～1.5 ratio mixing;
B) with the ethanol that adds entry or 40～60% in the said crude drug as extracting solvent, the solid-to-liquid ratio of said crude drug and said extraction solvent is 1: 5～10, extract more than 2 times, each 1.5～4.5 hours, filter merging filtrate afterwards, obtain extracting solution and residue;
C) be concentrated into said extracting solution dried; After add the water suspendible; Last macroporous resin; Adopt water, 30% ethanol, 60% ethanol and 95% ethanol to carry out continuous drip washing as eluent, obtain 30% ethanol drip washing thing respectively and be said the 1st extract, 60% ethanol drip washing thing is said the 2nd extract, 95% ethanol drip washing thing is said the 3rd extract;
D) said the 1st, 2,3 extracts are mixed, obtain said pharmaceutical composition.
2. method for preparing as claimed in claim 1, the weight ratio of Flos Lonicerae, Radix Scrophulariae, Radix Angelicae Sinensis, Radix Glycyrrhizae is 3: 3: 2 in the wherein said crude drug: 1.
3. method for preparing as claimed in claim 1, concentration of ethanol is 55% in the wherein said step b).
4. method for preparing as claimed in claim 1, solid-to-liquid ratio is 1: 6～8 in the wherein said step b).
5. method for preparing as claimed in claim 1, the extraction time in the wherein said step b) are 4 times, and each extraction time is 2～4 hours.
6. method for preparing as claimed in claim 1, wherein said macroporous resin are the AB-8 macroporous resin.
7. pharmaceutical composition as claimed in claim 1, wherein said the 1st extract, the 2nd extract, the 3rd extract are according to weight ratio 0.3～5.4: 0.2～4.2: 1 ratio is mixed.
8. pharmaceutical composition as claimed in claim 1, wherein said the 1st extract, the 2nd extract, the 3rd extract are according to weight ratio 0.6～3: 0.5～2.1: 1 ratio is mixed.
9. pharmaceutical composition as claimed in claim 1, wherein said the 1st extract, the 2nd extract, the 3rd extract mix according to 1.34: 1.05: 1 ratio of weight ratio.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102920838A (en) * | 2012-11-30 | 2013-02-13 | 哈药集团中药二厂 | Simiaoyong'an decoction dry paste powder and method for preparing oral preparation by using dry paste powder |
| CN103933207A (en) * | 2014-04-09 | 2014-07-23 | 滨州医学院附属医院 | Medicament for treating wegener granulomatosis with nose as first symptom and preparation method of medicament |
| CN116173115A (en) * | 2023-03-20 | 2023-05-30 | 北京中医药大学东直门医院 | Traditional Chinese medicine composition containing honeysuckle stem, application and preparation thereof |
| CN117838774A (en) * | 2022-12-23 | 2024-04-09 | 中国中医科学院中医基础理论研究所 | Formula medicine for preventing and treating neutrophil activity related diseases |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102920838A (en) * | 2012-11-30 | 2013-02-13 | 哈药集团中药二厂 | Simiaoyong'an decoction dry paste powder and method for preparing oral preparation by using dry paste powder |
| CN103933207A (en) * | 2014-04-09 | 2014-07-23 | 滨州医学院附属医院 | Medicament for treating wegener granulomatosis with nose as first symptom and preparation method of medicament |
| CN117838774A (en) * | 2022-12-23 | 2024-04-09 | 中国中医科学院中医基础理论研究所 | Formula medicine for preventing and treating neutrophil activity related diseases |
| CN116173115A (en) * | 2023-03-20 | 2023-05-30 | 北京中医药大学东直门医院 | Traditional Chinese medicine composition containing honeysuckle stem, application and preparation thereof |
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| CN102727654B (en) | 2015-06-10 |
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