CN104474068B - Fevervine extract and application thereof - Google Patents
Fevervine extract and application thereof Download PDFInfo
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- CN104474068B CN104474068B CN201410620120.1A CN201410620120A CN104474068B CN 104474068 B CN104474068 B CN 104474068B CN 201410620120 A CN201410620120 A CN 201410620120A CN 104474068 B CN104474068 B CN 104474068B
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- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
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Abstract
The present invention discloses fevervine extract, the extract contains iridoid glycosides compound, it is characterised in that the iridoid glycosides compound is mainly made up of 20.0% 50.0% Paederosidie Acid, 2.0% 25.0% paederoside and 1.0% 10.0% Paederosidie Acid methyl esters；The fevervine extract is prepared as follows：A) fevervine medicinal material extracts to obtain extract solution through ethanol；B) extract solution is concentrated into macroporous adsorption resin chromatography in no alcohol taste, successively with water and 80 90% ethanol elutions, collects eluent, is dried under reduced pressure, produces fevervine extract.The present invention also provides application of the described fevervine extract in anti-trioxypurine while the arthritic medicine of analgesic antigout is prepared.
Description
Technical field
The present invention relates to fevervine extract and application thereof.
Background technology
Fevervine is Rubiaceae Paederia plant fevervine Paederia scandens (Lour.) Merr. overground part
Point or herb, be Chinese traditional herbs, also known as Chinese Fevervine Herb, ox-hide jelly, relieving summer-heat rattan, horseshit rattan, smelly rattan, Sabia japonica etc., fevervine taste
It is thuja acid, mild-natured, enter liver, stomach, large intestine channel, there is the effect of promoting digestion and removing indigestion, expelling wind and activating blood flow, alleviating pain and detumescence.It is distributed in China and the southeast
Sub- multiple countries, the both at home and abroad pharmacological action to fevervine and fevervine clinical application research be concentrated mainly on anti-inflammatory, analgesia and
Treat disease of digestive system.Chemical constitution study in recent years shows to mainly contain the iridoid glycosides of sulfur-bearing in fevervine
Compound, there is the iridoid glycoside dimer of paederoside, Paederosidie Acid and a variety of sulfur-bearings.
CN101129524 is related to a kind of fevervine extract, and this application, which is used, contains ethanol (20%~95%) solvent extraction chicken
Rattan medicinal material is sweared, and points out to mainly contain Scandoside (10-90%) and Paederosidic acid methyl ester (10- in the extract of this method
90%).However, method disclosed in this application is cumbersome, high energy consumption.
The content of the invention
First purpose of the present invention is to provide fevervine extract, and the extract contains iridoid glycosides chemical combination
Thing, it is characterised in that the iridoid glycosides compound is mainly by 20.0%-50.0% Paederosidie Acid, 2.0%-
25.0% paederoside and 1.0%-10.0% Paederosidie Acids methyl esters composition；The fevervine extract is as follows
Prepare：
A) fevervine medicinal material extracts to obtain extract solution through ethanol；
B) extract solution is concentrated into macroporous adsorption resin chromatography in no alcohol taste, successively with water and 80-90% ethanol (preferably 85%
Ethanol) elution, eluent is collected, is dried under reduced pressure, produces fevervine extract.
According to a preferred embodiment, the ratio between the medicinal material and weight resin are that 1kg medicinal materials need 300ml through anhydrous second
Macroreticular resin after alcohol swelling pretreatment.
According to another preferred embodiment, the model D101 of the macroporous absorbent resin.
According to a preferred embodiment, the resin column blade diameter length ratio is 1: 8~1: 12.
According to another preferred embodiment, the loading flow velocity is 1.0~2.0BV/h, and elution flow rate is 4~6BV/
h。
According to the particularly preferred embodiment of the present invention, the preparation method bag of fevervine extract of the present invention
Include following specific steps：
Fevervine medicinal material W kg are taken, 8 times of 95% alcohol refluxs of amount is added and extracts 3 hours, filter extract solution, the dregs of a decoction add
8 times of 95% alcohol refluxs of amount extract 3 hours, filter extract solution, and the dregs of a decoction add 8 times of 95% alcohol refluxs of amount and extracted 3 hours, mistake
Extract solution is filtered, merges extract solution three times, 60 DEG C are concentrated under reduced pressure, and make to the deionized water solution without alcohol taste, 4 times of medicinal material amounts of addition molten
Solution, centrifuge, D101 macroporous absorbent resins in centrifuged supernatant, medicinal material is that 1kg medicinal materials need 300ml through absolute ethyl alcohol with amount of resin ratio
D101 macroreticular resins after swelling pretreatment, resin column blade diameter length ratio are 1: 8~1: 12, and loading flow velocity is 1.0~2.0BV/h, is washed
Separation of flow speed is 4~6BV/h.After end of the sample, deresination post 2BV is first washed with deionized water, discards and flows through liquid and water elution, then
With 85% ethanol-eluting resin column 6BV, the eluent of 85% ethanol is collected, 60 DEG C are concentrated under reduced pressure, and dry, and produce fevervine extraction
Thing.
Second object of the present invention is to provide fevervine extract and is preparing anti-trioxypurine analgesic antigout joint simultaneously
Purposes in scorching medicine.
Third object of the present invention is to provide use of the fevervine extract in the medicine for preparing treatment gouty arthritis
On the way.
Fourth object of the present invention is to provide use of the fevervine extract in the medicine for preparing treatment hyperuricemia
On the way.
The 5th purpose of the present invention is to provide fevervine extract and is preparing the use in preventing the medicine of uric acid mineralization
On the way.
Fevervine extract of the present invention can effectively reduce the serum uric acid level of high lithemia rat, while can also effectively change
Kind acetic acid causes the pain writhing response of mouse.In summary, fevervine extract of the invention has town while anti-trioxypurine
Pain activity.
The preparation method technique of the fevervine extract of the present invention is simple, reasonable in design, and environmental pollution is small.This method is to ring
Border is pollution-free, and ethanol consumption is few, and cost is relatively low, and yield is higher, suitable industrialized production.
Brief description of the drawings
Fig. 1 is the HPLC collection of illustrative plates of the gained fevervine extract of embodiment 1, wherein 1 represents Paederosidie Acid；2 represent chicken arrow
Rattan glycosides；3 represent Paederosidie Acid methyl esters；
Fig. 2 is the HPLC collection of illustrative plates of the gained fevervine extract of embodiment 2, wherein 1 represents Paederosidie Acid；2 represent chicken arrow
Rattan glycosides；3 represent Paederosidie Acid methyl esters.
Embodiment
The HPLC analysis method conditions of embodiment are as follows：
Mobile phase：Acetonitrile：1 ‰ formic acid waters, gradient elution, Detection wavelength 245nm, flow velocity 0.5ml/min, 30 DEG C of column temperature,
Chromatographic column Waters AtlantisTM C18,5u, 250 × 4.6mm.
Eluent gradient elutes ratio：
Using Paederosidie Acid as standard items, external standard method calculates Paederosidie Acid, paederoside and Paederosidie Acid methyl esters
Content.
Embodiment 1
Take Yunnan medicinal material (lot number：131110) 1.0kg, 8 times of 95% alcohol refluxs of amount is added and are extracted 3 hours, are extracted 3 times,
Filtering merges extract solution, and 60 DEG C are concentrated under reduced pressure, and to without alcohol taste, add deionized water makes dissolving to 4L, centrifuges, takes on supernatant
(medicinal material is with amount of resin than needing D101s of the 300ml after absolute ethyl alcohol swelling pretreatment big for 1kg medicinal materials for D101 macroporous absorbent resins
Hole resin), resin column blade diameter length ratio is 1: 8, and loading flow velocity is 1.0BV/h, after end of the sample, elution flow rate 5BV/h, is first spent
Ion water elution resin column 2BV, discards and flows through liquid and water elution, then with 85% ethanol-eluting resin column 6BV, collects 85% second
The eluent of alcohol, 60 DEG C are concentrated under reduced pressure, and dry, produce fevervine extract 16.43g, lot number 131110, yield 1.64%,
Using Paederosidie Acid as standard items external standard method calculate Paederosidie Acid, paederoside, Paederosidie Acid methyl esters content be respectively
27.33%th, 4.56%, 6.53%.
Embodiment 2
Take Yunnan medicinal material (lot number：131226) 1.0kg, 8 times of 95% alcohol refluxs of amount is added and are extracted 3 hours, are extracted 3 times,
Filtering merges extract solution, and 60 DEG C are concentrated under reduced pressure, and to without alcohol taste, add deionized water makes dissolving to 4L, centrifuges, takes on supernatant
(medicinal material is with amount of resin than needing D101s of the 300ml after absolute ethyl alcohol swelling pretreatment big for 1kg medicinal materials for D101 macroporous absorbent resins
Hole resin), resin column blade diameter length ratio is 1: 8, and loading flow velocity is 1.0BV/h, after end of the sample, elution flow rate 5BV/h, is first spent
Ion water elution resin column 2BV, discards and flows through liquid and water elution, then with 85% ethanol-eluting resin column 6BV, collects 85% second
The eluent of alcohol, 60 DEG C are concentrated under reduced pressure, and dry, produce fevervine extract 15.79g, lot number 131226, yield 1.58%,
Using Paederosidie Acid as standard items external standard method calculate Paederosidie Acid, paederoside, Paederosidie Acid methyl esters content be respectively
27.65%th, 18.23%, 2.20%.
Comparative example
The ethanol solution refluxing extraction that the fevervine herb volumetric concentration 1 kilogram is cleaned, crushed is 70% 2 hours,
Collect the ethanol solution and be concentrated under reduced pressure into dry, obtain about 100g medicinal extract.With the ethanol solution that 2 liters of volumetric concentrations are 10%
The medicinal extract of step 1 is dissolved at a temperature of 45 DEG C, then upper liquid will be taken after the solution centrifugal separation of gained.Gained supernatant is led to
Macroreticular resin (D101) chromatographic column is crossed, the weight of the macroreticular resin is 20 times of gained medicinal extract weight.It is with volumetric concentration
20% ethanol solution washes that this macroporous resin column is thin out to solution colour, and the ethanol consumption is the 10 of the macroreticular resin column volume
Times, flow velocity is 1 column volume/hour.This macroporous resin column to solution colour is washed with the ethanol solution that volumetric concentration is 60% again to become
Light, the ethanol consumption is 12 times of the macroreticular resin column volume, and flow velocity is 1 column volume/hour, collects eluent.Make to obtain
Eluent by being concentrated under reduced pressure dry, obtain fevervine extract 10.32g.
Test of pesticide effectiveness result
1.1 medicines and reagent
The fevervine extract lot number 131110 of embodiment 1；Comparative example.
Benzbromarone (the German big pharmaceutical factories of He Man, 1207062)；Glacial acetic acid (Chemical Reagent Co., Ltd., Sinopharm Group)；Yin
U.S. pungent enteric coatel tablets (the upper Hisense's friendship the Yellow River pharmaceutical Co. Ltd lot number of diindyl：A100101)；Uric acid (Uric acid) (U.S. Sigma
Company, lot number：LOT#BCBJ8674V)；Uric acid detection kit (Japanese Shino Test Corp., lot number YZB/
JAP 6068-2012)。
1.2 animal
Male ICR mouse, body weight 18-22g, by Shanghai, western pul-Bi Kai experimental animals Co., Ltd provides animal center
There is provided.Quality certification number：SCXK 2008-0016.
Male SD rat, body weight 130-150g, by Shanghai, western pul-Bi Kai experimental animals Co., Ltd provides animal center
There is provided.Quality certification number：SCXK 2008-0016.
1.3 instrument
Assay balance (Mettler Toledo Inc. of Switzerland, model：PL601-L)；Mouse claims that (Nanjing is with Ma Neili medicine instrument
Device Co., Ltd)；Full automatic serum Biochemical Analyzer (HIT, model：7080)；
1.4 method
(1) long-term prevention administration is to rat hyperuricemia model
70 rats are randomly divided into normal group, model group, fevervine extract doses difference group, positive drug group, every group 10
Only.The dosage group of fevervine extract is set to 25,50,100,200,400mgkgd-1, above medicine uses 0.5%CMC-
Na solution is configured to solution or suspension.Blank group and model group gavage physiological saline 1ml/d daily, the bodies such as remaining each group gavages
Product relative medicine, continuous gavage 7 days, one time a day.7th day, in addition to blank group, remaining each group 1 hour abdominal cavity note after gastric infusion
It is 250mg/kg uric acid suspension modelings to penetrate dosage.Rat eyeground vein clump intubation takes blood 0.5mL after 1 hour, and ice bath preserves, from
The heart, serum Full automatic serum Biochemical Analyzer in one hour detect uric acid content, the results are shown in Table 1.
(2) single therapy administration is to rat hyperuricemia model
33 rats are randomly divided into normal group, model group, fevervine extract group and comparative example group, every group 10.
Two administration group dosage are 400mgkgd-1, and above medicine is configured to solution or suspension with 0.5%CMC-Na solution
Liquid.Blank group and model group gavage physiological saline 1ml daily, and remaining each group gavages isometric relative medicine, remaining in addition to blank group
Each group 1 hour intraperitoneal injection dosage after gastric infusion is 250mg/kg uric acid suspension modelings.Rat eyeground vein after 1 hour
Clump intubation takes blood 0.5mL, and ice bath preserves, centrifugation, and serum Full automatic serum Biochemical Analyzer in one hour detects uric acid content,
It the results are shown in Table 2
(3) Dichlorodiphenyl Acetate causes the influence of mouse writhing
70 male indicated weight ICR mouse are randomly divided into model group, fevervine extract doses difference group, positive drug group, often
Group 10.Model group gives physiological saline, and positive controls give Indomethacin (10mgkgd-1), fevervine extract
Dosage group is set to 50,100,200,400 and 800mgkgd-1, above medicine is configured to solution with 0.5%CMC-Na solution
Or suspension.After last dose 60min, 0.7% acetum 0.1mL/10g is injected intraperitoneally, with mouse web portion indent, hind leg
Stretch, buttocks is lifted as index, records mouse writhing reaction times in 15min, and is calculated analgesia and suppressed percentage, the results are shown in Table
3。
Ease pain percentage %=(control group writhing mean-administration group writhing mean)/control group writhing mean × 100%.
All data carry out statistical disposition with SPSS20.0 softwares, withRepresent, examined using T.
1.5 result
(1) long-term influence of the prevention administration to hyperuricemia rat blood serum uric acid：As shown in Table 1, hyperuricemia mould
Type group rat blood serum uric acid level is apparently higher than normal group (P ＜ 0.01), compared with model group, when positive drug gives 50mg/kg
When can significantly decrease the serum uric acid level (P ＜ 0.05) of hyperuricemia rat；When fevervine extract gives 50,100,
200th, the serum uric acid level (P ＜ 0.05,0.01) of hyperuricemia rat can be significantly decreased during 400mg/kg, and is in concentration
Dependence.
Influence of the 1 long-term prevention administration of table to rat hyperuricemia
Compared with normal group：##P ＜ 0.01；Compared with model group：* P ＜ 0.05, * * P ＜ 0.01
(2) influence to hyperuricemia rat blood serum uric acid is administered in single therapy：As shown in Table 2, hyperuricemia mould
Type group rat blood serum uric acid level is apparently higher than normal group (P ＜ 0.01), compared with model group, when comparative example is given
During 400mg/kg, rat blood uric acid is without significantly sexually revising；Height can be significantly decreased when fevervine extract gives 400mg/kg
The serum uric acid level (P ＜ 0.05) of uricacidemia rat.Illustrate that the effect of fevervine extract anti-trioxypurine is better than comparative example.
The single therapy of table 2 be administered to the shadow of rat hyperuricemia to
Compared with normal group：##P ＜ 0.01；Compared with model group：* P ＜ 0.05
(3) influence of mouse writhing caused by Dichlorodiphenyl Acetate：As shown in table 3, compared with model group, Indomethacin is given
10mg/kg can effectively suppress the mouse writhing reaction (P ＜ 0.001) of acetic-acid induced；EPS various doses be able to can be reduced effectively
Mouse writhing number (50mg/kg, the P ＜ 0.01 of acetic-acid induced in 15min；100mg/kg, P ＜ 0.001,200mg/kg, P ＜
0.001；400mg/kg, P ＜ 0.001；800mg/kg, P ＜ 0.001)；Inhibiting rate is respectively 46.4%, 65.2%, 68.0%,
Inhibiting rate difference is little between 76.2% and 74.9%, 100-800mg/kg dosage, there is obvious dose dependent.
Mouse writhing number caused by the Dichlorodiphenyl Acetate of table 3 influence (N=10)
Compared with model group*P ＜ 0.05,**P ＜ 0.01,***P ＜ 0.001
Hyperuricemia is the important biochemical basis of urarthritis, maintains normal serum uric acid level, for anti-
Only uric acid mineralization has important biochemical basis meaning, meanwhile, pain caused by inflammation is the main for the treatment of urarthritis
One of therapeutic purpose.Effect experiment result of the present invention shows that the fevervine extract of present invention gained can effectively reduce height
The serum uric acid level of uric acid rat, while also can effectively improve the pain writhing response that acetic acid causes mouse.In summary, it is of the invention
Fevervine extract has analgesic activities while anti-trioxypurine.
Claims (1)
1. purposes of the fevervine extract in anti-trioxypurine while the arthritic medicine of analgesic antigout is prepared, the extract
Contain iridoid glycosides compound, it is characterised in that the iridoid glycosides compound is mainly by 20.0%-50.0%'s
Paederosidie Acid, 2.0%-25.0% paederoside and 1.0%-10.0% Paederosidie Acids methyl esters composition；
The fevervine extract is prepared as follows：
A) fevervine medicinal material extracts to obtain extract solution through ethanol；
B) extract solution is concentrated into macroporous adsorption resin chromatography in no alcohol taste, successively with water and 85% ethanol elution, collects eluent,
It is dried under reduced pressure, produces fevervine extract；
The ratio between the medicinal material and weight resin are that 1kg medicinal materials need macroreticular resins of the 300ml after absolute ethyl alcohol swelling pretreatment；
The model D101 of the macroporous absorbent resin；
The resin column blade diameter length ratio is 1:8~1:12；
The loading flow velocity is 1.0~2.0BV/h, and elution flow rate is 4~6BV/h.
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| CN104435226A (en) * | 2014-11-06 | 2015-03-25 | 宁波大昌药业有限公司 | Paederia scandens extract and application thereof |
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| CN101890033A (en) * | 2009-05-20 | 2010-11-24 | 苏州颐华生物医药技术有限公司 | Application of paederosidic acid used for preparing analgesic drugs and/or anti-inflammatory drugs |
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| CN104435226A (en) * | 2014-11-06 | 2015-03-25 | 宁波大昌药业有限公司 | Paederia scandens extract and application thereof |
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