CN103417644B - A kind of Heiguteng exract compound extract and its production and use - Google Patents
A kind of Heiguteng exract compound extract and its production and use Download PDFInfo
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Abstract
The invention provides the preparation method of Heiguteng exract compound extract, it comprises following operating procedure: (1) is by compound recipe weight proportion weighting raw materials: Heiguteng exract 2.5 ~ 10 parts, Radix Gentianae Macrophyllae 1 ~ 4 part, Rhizoma Corydalis 1 ~ 4 part; After 40 ~ 60%v/v ethanol extraction, merge extractive liquid, reclaims ethanol to without after alcohol taste, obtains crude extract; (2) get crude extract, upper non-polar macroporous resin post, after ion, with 20 ~ 60%v/v ethanol elution, collect ethanol elution, after concentrated, drying, obtain Heiguteng exract compound extract.Present invention also offers a kind of Heiguteng exract compound extract and uses thereof.The present invention's research shows, the Heiguteng exract compound extract obtained with preparation method of the present invention has good drug activity to rheumatoid arthritis, and effect is better than other extract parts, specify that the effective site of Heiguteng exract compound recipe further, for preparation and drug safety monitoring provide guarantee.
Description
Technical field
The present invention relates to a kind of Heiguteng exract compound extract and its production and use.
Background technology
Rheumatoid arthritis (RheumatioidArthritis, RA) be a kind of autoimmune disorder disease, with synovium of joint chronic inflammatory disease for principal character, " arthromyodynia " category is belonged in Chinese medicine, concrete pathogenic factor it be unclear that, and it is generally acknowledged relevant with many factors such as environment, infection, immune disorders.RA is about 0.4% at China's sickness rate, and focus mostly on southern province.Therapeutic Method has drug treatment and the physical therapy modalities such as chemical medicine, Chinese medicine, biological preparation.
Heiguteng exract is dry root or the Herb of Asclepiadaceae (Asclepiasaceae) Periploca (PeriplocaLinn.) plant Caulis et Radix Periplocae Forrestii (PeriplocaforrestiiSchltr), having effect of stasis-dispelling and pain-killing, expelling wind and removing dampness, dredge the meridian passage, is the common medicine being widely used in treating rheumatism and the disease such as rheumatoid, closed soft tissue injury among the people.
Radix Gentianae Macrophyllae is the dry root of Gentianaceae Radix Gentianae Macrophyllae (GentianamacrophyllaPall), gentiana straminea maxim (G.stramineaMaxim), gentiana crassicaulis Duthie (G.crassicaulisDuthieexBurk) or Gentiana dahurica (G.dahuricaFisch), has to protect the liver, treat the effects such as rheumatic arthritis, clearing away damp-heat, antiallergic, stopping numbness pain.
Rhizoma Corydalis has another name called Rhizoma Corydalis, is the dry tuber of papaveraceae plant corydalis (CorydalisyanhusuoW.T.Wang).There is effect of blood circulation promoting and blood stasis dispelling, regulating QI to relieve pain.
Number of patent application: 201210094512.x, disclose a kind of pharmaceutical composition for the treatment of rheumatoid arthritis, formula is Heiguteng exract 2.5 ~ 10 parts, Radix Gentianae Macrophyllae 1 ~ 4 part, Rhizoma Corydalis 1 ~ 4 part, test shows, 50% ethanol extraction of above-mentioned compound recipe effectively can treat rheumatoid arthritis, and toxic and side effects is low, containing total flavones 2-4%, total steroid saponin 3 ~ 6%, total alkaloids 0.1 ~ 0.3%, gentiopicrin 1 ~ 3%, tetrahydropalmatine 0.04 ~ 0.06%, periplocoside 0.1 ~ 0.2% in its ethanol extraction.
In above-mentioned patent, the drug activity of compound recipe is better, but in its extract, active constituent content is lower, if be prepared into square preparation, supplementary product consumption is comparatively large, is unfavorable for the saving of commercial production cost, and its dosage is comparatively large, is not easy to patient and uses.If a kind of active constituent content can be provided higher and the significant compound extract of drug activity, the popularization of above-mentioned compound medicine will be conducive to.
Summary of the invention
The object of the present invention is to provide a kind of Heiguteng exract compound extract and its production and use.
The invention provides the preparation method of Heiguteng exract compound extract, it comprises following operating procedure:
(1) by compound recipe weight proportion weighting raw materials: Heiguteng exract 2.5 ~ 10 parts, Radix Gentianae Macrophyllae 1 ~ 4 part, Rhizoma Corydalis 1 ~ 4 part; After 40 ~ 60%v/v ethanol extraction, merge extractive liquid, reclaims ethanol to without after alcohol taste, obtains crude extract;
(2) get crude extract, upper non-polar macroporous resin post, after ion, with 20 ~ 60%v/v ethanol elution, collect ethanol elution, after concentrated, drying, obtain Heiguteng exract compound extract.
Further, compound recipe weight proportion is: Heiguteng exract 2.5 parts, Radix Gentianae Macrophyllae 1 part, Rhizoma Corydalis 1 part.
Further, in step (1), described concentration of alcohol is 50%v/v.
Further, in step (2), described non-polar macroporous resin is D101.
Further, in step (2), concrete elution action is one of following scheme:
Option A: after ion, uses 20%v/v ethanol elution, collects ethanol elution;
Option b: after ion, uses 20%v/v, 40%v/v ethanol elution successively, collects 40%v/v ethanol elution;
Scheme C: after ion, uses 40%v/v, 60%v/v ethanol elution successively, collects 60%v/v ethanol elution.
Wherein, in scheme C, before 40%v/v ethanol elution, first use 20%v/v ethanol elution.
Present invention also offers Heiguteng exract compound extract prepared by said method, during option A is extract obtained, gentiopicroside in different morphological is 7 ~ 9%w/w, periplocoside content is 0 ~ 1%w/w, Content determination of dl-tetrahydropalmatine is 0 ~ 1%w/w, and general flavone content is 11 ~ 15%w/w, and total saponin content is 7 ~ 9%w/w;
During option b is extract obtained, gentiopicroside in different morphological is 0.4 ~ 0.6%w/w, and periplocoside content is 3 ~ 5%w/w, and Content determination of dl-tetrahydropalmatine is 0 ~ 1%w/w, and general flavone content is 16 ~ 19%w/w, and total saponin content is 15 ~ 18%w/w;
During scheme C is extract obtained, gentiopicroside in different morphological is 0.01 ~ 0.2%w/w, and periplocoside content is 1 ~ 3%w/w, and Content determination of dl-tetrahydropalmatine is 2 ~ 4%w/w, and general flavone content is 8 ~ 12%w/w, and total saponin content is 15 ~ 18%w/w.
Further, during option A is extract obtained, gentiopicroside in different morphological is 8%w/w, and periplocoside content is 0%w/w, and Content determination of dl-tetrahydropalmatine is 0%w/w, and general flavone content is 13 ~ 14%w/w, and total saponin content is 8 ~ 9%w/w;
During option b is extract obtained, gentiopicroside in different morphological is 0.5 ~ 0.6%w/w, and periplocoside content is 4%w/w, and Content determination of dl-tetrahydropalmatine is 0%w/w, and general flavone content is 17 ~ 18%w/w, and total saponin content is 16 ~ 17%w/w;
During scheme C is extract obtained, gentiopicroside in different morphological is 0.1 ~ 0.2%w/w, and periplocoside content is 2%w/w, and Content determination of dl-tetrahydropalmatine is 2 ~ 3%w/w, and general flavone content is 10 ~ 11%w/w, and total saponin content is 16 ~ 18%w/w.
Present invention also offers the purposes of above-mentioned Heiguteng exract compound extract in the medicine preparing antiinflammatory or treatment rheumatoid arthritis.
Present invention also offers a kind of pharmaceutical composition for the treatment of rheumatoid arthritis, it be with the extract obtained a kind of or option A of above-mentioned option A ~ C and the extract obtained compositions of option b for active component, add the preparation that pharmaceutically conventional adjuvant or complementary composition prepare.
In option A and the extract obtained compositions of option b, both proportionings are 1:1w/w.
Further, described preparation is oral agents, such as tablet, capsule, pill, powder, granule etc.
The present invention's research shows, the Heiguteng exract compound extract obtained with preparation method of the present invention has good drug activity to rheumatoid arthritis, and effect is better than other extract parts, specify that the effective site of Heiguteng exract compound recipe, for further preparation and drug safety monitoring provide guarantee.
Detailed description of the invention
The preparation of embodiment 1 Heiguteng exract compound extract of the present invention
Prescription:
Heiguteng exract 50g, Radix Gentianae Macrophyllae 20g, Rhizoma Corydalis 20g
Preparation method:
(1) weighting raw materials by weight, after 50%v/v ethanol ultrasonic extraction, merge extractive liquid, reclaims ethanol to without after alcohol taste, obtains crude extract;
(2) get crude extract, upper D101 macroporous resin column, first wash remove impurity (20 times of column volumes) with water, then use 20 times of column volume 20%v/v ethanol elutions, collect ethanol elution, after concentrating under reduced pressure, lyophilizing, obtains Heiguteng exract compound extract.
The assay method of total flavones, total saponins in the present invention, the ultraviolet spectrophotometry can commonly used see this area; The detection method of gentiopicrin, periplocoside and tetrahydropalmatine can see Chinese Patent Application No.: 201210094512.x.
The preparation of embodiment 2 Heiguteng exract compound extract of the present invention
Prescription:
Heiguteng exract 50g, Radix Gentianae Macrophyllae 20g, Rhizoma Corydalis 20g
Preparation method:
(1) weighting raw materials by weight, after 50%v/v ethanol ultrasonic extraction, merge extractive liquid, reclaims ethanol to without after alcohol taste, obtains crude extract;
(2) get crude extract, upper D101 macroporous resin column, first wash remove impurity (20 times of column volumes) with water, use 20 times of column volume 20%v/v, 40%v/v, 60%v/v ethanol elutions more respectively successively, collect 60% ethanol elution, after concentrating under reduced pressure, lyophilizing, obtains Heiguteng exract compound extract.
The preparation of embodiment 3 Heiguteng exract compound extract of the present invention
Prescription:
Heiguteng exract 50g, Radix Gentianae Macrophyllae 20g, Rhizoma Corydalis 20g
Preparation method:
(1) weighting raw materials by weight, after 50%v/v ethanol ultrasonic extraction, merge extractive liquid, reclaims ethanol to without after alcohol taste, obtains crude extract;
(2) get crude extract, upper D101 macroporous resin column, first wash remove impurity (20 times of column volumes) with water, use 20 times of column volume 20%v/v, 40%v/v ethanol elutions more respectively successively, collect 40% ethanol elution, after concentrating under reduced pressure, lyophilizing, obtains Heiguteng exract compound extract.
The preparation of embodiment 4 Heiguteng exract compound extract of the present invention
Prescription:
Heiguteng exract 50g, Radix Gentianae Macrophyllae 20g, Rhizoma Corydalis 20g
Preparation method:
(1) weighting raw materials by weight, after 50%v/v ethanol ultrasonic extraction, merge extractive liquid, reclaims ethanol to without after alcohol taste, obtains crude extract;
(2) get crude extract, upper D101 macroporous resin column, first wash remove impurity (20 times of column volumes) with water, then use 20 times of column volume 40%v/v ethanol elutions, collect 40% ethanol elution, after concentrating under reduced pressure, lyophilizing, obtains Heiguteng exract compound extract.
The preparation of embodiment 5 pharmaceutical composition of the present invention
Heiguteng exract compound extract prepared by Example 1 or embodiment 2, adds appropriate amount of starch, dextrin, granulates, obtain granule.
The preparation of embodiment 6 pharmaceutical composition of the present invention
Heiguteng exract compound extract prepared by Example 1 and embodiment 3, after 1:1w/w mixing, add appropriate amount of starch, dextrin, granulate, tabletting, obtains tablet.
The extract that the different eluate concentration of embodiment 7 prepares
The preparation of 1 crude extract
Get Heiguteng exract 500g, Radix Gentianae Macrophyllae 200g, Rhizoma Corydalis 200g, after pulverizing, mixing, after adding 50%v/v ethanol ultrasonic extraction, merge extractive liquid, after reclaiming ethanol, concentrated, lyophilizing, obtains purification crude extract.
2 eluate concentration are on the impact of purification effect
Take clean macroporous resin D1012kg to chromatographic column dress post, take Heiguteng exract crude extract dry powder 200g and add loading after 1000ml distilled water ultrasonic dissolution, be each 20 times of column volume eluting of alcoholic solution of 0%, 20%, 40%, 60%, 80%, 95% successively by concentration, after collecting each eluent spin concentration, lyophilizing is for subsequent use, measures the content of total saponins, total flavones, periplocoside, tetrahydropalmatine, gentiopicrin.Detection method is with reference to prior art.
2.1 gentiopicrins, periplocoside and Content determination of dl-tetrahydropalmatine
Result sees table.
In each concentration ethanol solution of table 1 gentiopicrin, periplocoside and tetrahydropalmatine content (
n=3)
Wherein A group: 0% ethanol elution; B group: 20% ethanol elution; C group: 40% ethanol elution; D group: 60% ethanol elution; E group: 80% ethanol elution; F group: 95% ethanol elution; G group: sample before purification.
As seen from the above table, the bitter glycosides of gallbladder content in B group is the highest, and enrichment times is about 7 times; Periplocoside content in C group is the highest, and enrichment times is about 14 times; Tetrahydropalmatine content in D group is the highest, and enrichment times is about 24 times.General effect is obvious.
2.2 total flavones and total saponin content
The results are shown in following table.
The content of table 2 total flavones and total saponins (
n=3)
Wherein A group: 0% ethanol elution; B group: 20% ethanol elution; C group: 40% ethanol elution; D group: 60% ethanol elution; E group: 80% ethanol elution; F group: 95% ethanol elution; G group: sample before purification.
Total flavones is the highest at B group content, and total saponins is the highest at C group content, and total flavones and total saponins all reach concentration effect in B, C, D, E group.
Beneficial effect of the present invention is illustrated below by way of test example.
Test example 1 effect experiment
The mice ear experiment of 1.1 each extract parts
1.1.1 laboratory animal
Regular grade Kunming is healthy mouse, body weight 18 ~ 22g, 60 ages in days, Quan Xiong, sub-cage rearing, reaches large animal feeding center purchased from Chengdu.
1.1.2 experiment material, reagent and instrument
Sample after each gradient eluent drying in embodiment 7.
Positive control drug prednisone (the celestial jade pendant Pharmaceutical in Zhejiang), sodium carboxymethyl cellulose, dimethylbenzene are purchased from Chengdu Ke Long chemical reagent factory, and water is distilled water.
ESJ120-4 ten thousand/electronic balance (Shenyang Long Teng), operating scissors, card punch, tweezers.
1.1.3 Pharmaceutical setting method
Take 5g sodium carboxymethyl cellulose in 1000mL beaker, adding distil water heating for dissolving, after being cooled to room temperature, polishing distilled water to graduation mark obtains 0.5% carboxymethylcellulose sodium solution 100mL, and tablet dry powder to the 100mL volumetric flask taken containing 25mg prednisolone acetate adds 0.5% sodium carboxymethyl cellulose and dissolves and obtain 0.25mg/ml prednisolone acetate solution 100mL.
Take 20% respectively, 40%, in 60%v/v ethanol elution product 1.25g to 50mL volumetric flask, then add respectively 0.5% sodium carboxymethyl cellulose dissolve obtain concentration be 25mg/ml 20%, 40%, 60%v/v eluted product solution 50mL.
Pipette the 40%v/v ethanol elution reaction mixture mixing that 20%v/v ethanol elution reaction mixture that 10mL concentration is 25mg/mL and 10mL concentration are 25mg/mL, obtain 20% and 40% ethanol elution product mixed solution.
Pipette the 60%v/v ethanol elution reaction mixture mixing that 20%v/v ethanol elution reaction mixture that 10mL concentration is 25mg/mL and 10mL concentration are 25mg/mL, obtain 20% and 60%v/v ethanol elution product mixed solution.
Pipette the 60%v/v ethanol elution reaction mixture mixing that 40%v/v ethanol elution reaction mixture that 10mL concentration is 25mg/mL and 10mL concentration are 25mg/mL, obtain 40% and 60%v/v ethanol elution product mixed solution.
1.1.4 mice ear experiment
After buying 18-22g healthy mice, adaptability feeds three days, is divided into 8 groups immediately, often organizes 10.
A group is blank group, gavage 0.5% carboxymethylcellulose sodium solution, each 0.2mL.
B group is positive group, gavage 0.25mg/ml prednisolone acetate solution, each 0.2mL.
C group is 20% ethanol group, gavage 25mg/mL20%v/v ethanol elution reaction mixture, each 0.2mL.
D group is 40% ethanol group, gavage 25mg/mL40%v/v ethanol elution reaction mixture, each 0.2mL.
E group is 60% ethanol group, gavage 25mg/mL60%v/v ethanol elution reaction mixture, each 0.2mL.
F group is 20% and 40% ethanol mixing group, gavage 25mg/mL20% and 40%v/v ethanol elution product mixed solution, each 0.2mL.
G group is 20% and 60% ethanol mixing group, gavage 25mg/mL20% and 60%v/v ethanol elution product mixed solution, each 0.2mL.
H group is 40% and 60% ethanol mixing group, gavage 25mg/mL40% and 60%v/v ethanol elution product mixed solution, each 0.2mL.
Every 12 hours gavages once, continuous gavage 6 times.Last administration evenly coats dimethylbenzene to after 1 hour mouse right ear, and after half an hour, disconnected vertebra is put to death, and cuts left and right two ears, the large auricle such as to lay weigh with card punch at left and right two ear same position.
Computing formula is as follows:
1.2 experimental result
1.3.1 mice ear experimental result
The results are shown in Table 3 through SPSS13.0 analysis design mothod.
The experiment of table 3 mice ear (
n=10)
Note: *: contrast P<0.05** with model group: contrast P<0.01 with model group
Table 3 result shows, administration group C, F swelling rate and model group contrast pole significant difference, has antiphlogistic effects.Experimental result shows that administration group D, E swelling rate and model group contrast and have significant difference, have good antiphlogistic effects.As can be seen here, the drug action of 20%v/v alcohol elution is best, is secondly 40%, 60%v/v alcohol elution; Test simultaneously and also find, after 20%, 40% alcohol elution mixing, its drug action have also been obtained enhancing, and after 20% and 60%, 40% and 60% alcohol elution mixing, drug activity reduces on the contrary.
The present invention's research shows, the Heiguteng exract compound extract obtained with preparation method of the present invention has good drug activity to rheumatoid arthritis, and effect is better than other extract parts, specify that the effective site of Heiguteng exract compound recipe further, for preparation and drug safety monitoring provide guarantee.
Claims (10)
1. the preparation method of Heiguteng exract compound extract, is characterized in that: it comprises following operating procedure:
(1) by compound recipe weight proportion weighting raw materials: Heiguteng exract 2.5 parts, Radix Gentianae Macrophyllae 1 part, Rhizoma Corydalis 1 part; After 40 ~ 60%v/v ethanol extraction, merge extractive liquid, reclaims ethanol to without after alcohol taste, obtains crude extract;
(2) get crude extract, upper non-polar macroporous resin post, after ion, with 20 ~ 60%v/v ethanol elution, collect ethanol elution, after concentrated, drying, obtain Heiguteng exract compound extract; Described non-polar macroporous resin is D101;
Concrete elution action is one of following scheme:
Option A: after ion, uses 20%v/v ethanol elution, collects ethanol elution;
Option b: after ion, uses 20%v/v, 40%v/v ethanol elution successively, collects 40%v/v ethanol elution;
Scheme C: after ion, uses 40%v/v, 60%v/v ethanol elution successively, collects 60%v/v ethanol elution.
2. preparation method according to claim 1, is characterized in that: in step (1), and described concentration of alcohol is 50%v/v.
3. preparation method according to claim 1, is characterized in that: in scheme C, before 40%v/v ethanol elution, first uses 20%v/v ethanol elution.
4. the Heiguteng exract compound extract prepared of method described in claim 1-3 any one, it is characterized in that: during option A is extract obtained, gentiopicroside in different morphological is 7 ~ 9%w/w, periplocoside content is 0 ~ 1%w/w, Content determination of dl-tetrahydropalmatine is 0 ~ 1%w/w, general flavone content is 11 ~ 15%w/w, and total saponin content is 7 ~ 9%w/w;
During option b is extract obtained, gentiopicroside in different morphological is 0.4 ~ 0.6%w/w, and periplocoside content is 3 ~ 5%w/w, and Content determination of dl-tetrahydropalmatine is 0 ~ 1%w/w, and general flavone content is 16 ~ 19%w/w, and total saponin content is 15 ~ 18%w/w;
During scheme C is extract obtained, gentiopicroside in different morphological is 0.01 ~ 0.2%w/w, and periplocoside content is 1 ~ 3%w/w, and Content determination of dl-tetrahydropalmatine is 2 ~ 4%w/w, and general flavone content is 8 ~ 12%w/w, and total saponin content is 15 ~ 18%w/w.
5. Heiguteng exract compound extract according to claim 4, is characterized in that:
During option A is extract obtained, gentiopicroside in different morphological is 8%w/w, and periplocoside content is 0%w/w, and Content determination of dl-tetrahydropalmatine is 0%w/w, and general flavone content is 13 ~ 14%w/w, and total saponin content is 8 ~ 9%w/w;
During option b is extract obtained, gentiopicroside in different morphological is 0.5 ~ 0.6%w/w, and periplocoside content is 4%w/w, and Content determination of dl-tetrahydropalmatine is 0%w/w, and general flavone content is 17 ~ 18%w/w, and total saponin content is 16 ~ 17%w/w;
During scheme C is extract obtained, gentiopicroside in different morphological is 0.1 ~ 0.2%w/w, and periplocoside content is 2%w/w, and Content determination of dl-tetrahydropalmatine is 2 ~ 3%w/w, and general flavone content is 10 ~ 11%w/w, and total saponin content is 16 ~ 18%w/w.
6. the purposes of Heiguteng exract compound extract in the medicine preparing antiinflammatory described in claim 4 or 5.
7. the purposes of Heiguteng exract compound extract described in claim 4 or 5 in the medicine of preparation treatment rheumatoid arthritis.
8. an anti-inflammatory pharmaceutical compositions, is characterized in that: it be with the extract obtained one of the option A ~ C described in claim 4 or 5 for active component, add the preparation that pharmaceutically conventional adjuvant prepares.
9. an anti-inflammatory pharmaceutical compositions, is characterized in that: it be with the option A described in claim 4 or 5 and the extract obtained compositions of option b for active component, add the preparation that pharmaceutically conventional adjuvant prepares.
10. pharmaceutical composition according to claim 9, is characterized in that: in described option A and the extract obtained compositions of option b, both proportionings are 1:1w/w.
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