CN105434567A - Traditional Chinese medicine composition for treating rheumatoid arthritis and preparation method thereof - Google Patents

Traditional Chinese medicine composition for treating rheumatoid arthritis and preparation method thereof Download PDF

Info

Publication number
CN105434567A
CN105434567A CN201511019368.3A CN201511019368A CN105434567A CN 105434567 A CN105434567 A CN 105434567A CN 201511019368 A CN201511019368 A CN 201511019368A CN 105434567 A CN105434567 A CN 105434567A
Authority
CN
China
Prior art keywords
radix aconiti
aconiti lateralis
lateralis preparata
rhizoma atractylodis
atractylodis macrocephalae
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201511019368.3A
Other languages
Chinese (zh)
Other versions
CN105434567B (en
Inventor
徐攀
浦锦宝
胡轶娟
梁卫青
程林
张宏建
周洁
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Zhejiang Traditional Chinese Medicine Research Institute
Zhejiang Academy of Traditional Chinese Medicine
Original Assignee
Zhejiang Traditional Chinese Medicine Research Institute
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Zhejiang Traditional Chinese Medicine Research Institute filed Critical Zhejiang Traditional Chinese Medicine Research Institute
Priority to CN201511019368.3A priority Critical patent/CN105434567B/en
Publication of CN105434567A publication Critical patent/CN105434567A/en
Application granted granted Critical
Publication of CN105434567B publication Critical patent/CN105434567B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/71—Ranunculaceae (Buttercup family), e.g. larkspur, hepatica, hydrastis, columbine or goldenseal
    • A61K36/714—Aconitum (monkshood)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
    • A61K36/284—Atractylodes
    • A—HUMAN NECESSITIES
    • 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
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
    • A—HUMAN NECESSITIES
    • 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
    • A61K2236/30—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
    • A—HUMAN NECESSITIES
    • 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
    • A61K2236/30—Extraction of the material
    • A61K2236/35—Extraction with lipophilic solvents, e.g. Hexane or petrol ether
    • A—HUMAN NECESSITIES
    • 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
    • A61K2236/30—Extraction of the material
    • A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
    • A—HUMAN NECESSITIES
    • 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
    • A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
    • A—HUMAN NECESSITIES
    • 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

Landscapes

  • Health & Medical Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Microbiology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Biotechnology (AREA)
  • Botany (AREA)
  • Medical Informatics (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Mycology (AREA)
  • Engineering & Computer Science (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention relates to a traditional Chinese medicine composition for treating rheumatoid arthritis and a preparation method thereof. The traditional Chinese medicine composition is prepared from the following extracts: 15-35% of radix aconiti carmichaeli and rhizoma atractylodis macrocephalae polysaccharide, 23-43% of rhizoma atractylodis macrocephalae lactone and 30-50% of radix aconiti carmichaeli alkaloid. The preparation method comprises the following steps: 1), preparation of radix aconiti carmichaeli and rhizoma atractylodis macrocephalae polysaccharide: weighing medicinal materials namely radix aconiti carmichaeli and rhizoma atractylodis macrocephalae, extracting the medicinal materials with petroleum ether, and then extracting with ethyl alcohol for removing impurities, extracting medicine dregs with water so as to obtain a concentrated medical solution, adding absolute ethyl alcohol in the concentrated medical solution, carrying out alcohol precipitation, carrying out alcohol precipitation again after protein is removed, sequentially washing with absolute ethyl alcohol and acetone, and carrying out vacuum drying so as to obtain the radix aconiti carmichaeli and rhizoma atractylodis macrocephalae polysaccharide; 2), preparation of rhizoma atractylodacrocephalaeis m lactone and radix aconiti carmichaeli alkaloid: after refluxing is carried out with ethyl alcohol in a water bath, centrifuging for removing insoluble matters, loading medical solutions to the upper end of a macroporous resin column for eluting with pure water and ethyl alcohol in sequence, collecting eluents and carrying out vacuum drying so as to obtain the rhizoma atractylodis macrocephalae lactone and the radix aconiti carmichaeli alkaloid. The traditional Chinese medicine composition is better applied on the aspect of medicines for treating rheumatoid arthritis.

Description

A kind of Chinese medicine composition for the treatment of rheumatoid arthritis and preparation method thereof
Technical field
The present invention relates to the field of Chinese medicines, be specifically related to the active component Radix Aconiti Lateralis Preparata total alkaloids extracted from Radix Aconiti Lateralis Preparata, the composition and method of making the same of the active component atractylodes lactone extracted from the Rhizoma Atractylodis Macrocephalae and the attached art polysaccharide of the active component extracted from the Radix Aconiti Lateralis Preparata Rhizoma Atractylodis Macrocephalae.
Background technology
Rheumatoid arthritis (rheumatoidarthritis, RA) is a kind of common autoimmune disease being the chronic inflammation of feature with Progressive symmetric erythrokeratodermia articular cartilage and bone damage, and finally can cause joint deformity and afunction, disability rate is high.According to latest survey display, the sickness rate of China's rheumatoid arthritis is very high, and number of patients is about more than 3,000 ten thousand, and in adult, sickness rate can up to about 2%.It is the one of " arthromyodynia " in rheumatoid arthritis Chinese medical theory system in ancient times, Chinese medicine arthromyodynia has the history of several thousand, in " Treatise on Febrile and Miscellaneous Disease ", " Medical Treasures of the Golden Chamber " of Huangdi's Internal Classics and Zhang Zhongjing, just the discussion of system is carried out to the aspect such as the cause of disease, pathogenesis, sick position, syndrome, prognosis of arthromyodynia." Radix Aconiti Lateralis Preparata-Rhizoma Atractylodis Macrocephalae " medicine, to the classical prescription coming from Zhang Zhongjing, is also the herbal pair that modern clinic is used for the treatment of rheumatoid arthritis.Radix Aconiti Lateralis Preparata, dispelling cold by warming the meridian can pass through 12 warps, Rhizoma Atractylodis Macrocephalae dampness, and that closes then has dispelling cold and dampness, the meridian dredging, the merit of stopping numbness pain, and be one of characteristic compatibility of Zhong Jingfang treatment " arthromyodynia ", both complementations are use, embody the principles of formulating prescriptions of differential diagnosis in tcm.Research shows, Radix Aconiti Lateralis Preparata and Rhizoma Atractylodis Macrocephalae compatibility effectively can suppress the arthroncus of adjuvant arthritis rats Secondary cases, significantly can reduce the level of adjuvant arthritis rats serum NO level and TNF-α.
And the research right to Radix Aconiti Lateralis Preparata Rhizoma Atractylodis Macrocephalae medicine at present is mainly, by experimental animal model, carry out the pharmacological effect before and after compatibility of drugs and pharmaceutical chemistry research, thus science evidence or checking are carried out to traditional compatibility, make it possess scientific basis or carry out developing new drug for clinical application by the experimentation of compatibility and rational experimental basis is provided.In addition, also studied by the dosage ratio of different proportion, disclose the dose ratio that it plays the best use of.The research method of above Radix Aconiti Lateralis Preparata Rhizoma Atractylodis Macrocephalae Compatibility Law rests on Chinese medicine itself generally substantially, and a Chinese medicine inherently complicated system, there is complicated chemical composition and drug action, therefore with the main active substances in Chinese medicine compound for object of study, study the proportion compatibility of its active substance and to carry out the secondary development of medicine based on this more reasonable.
Summary of the invention
First object of the present invention is to provide a kind of Chinese medicine composition for the treatment of rheumatoid arthritis, and second object of the present invention is to provide a kind of preparation method for the treatment of the Chinese medicine composition of rheumatoid arthritis.
In order to realize above-mentioned first object, present invention employs following technical scheme:
Treat a Chinese medicine composition for rheumatoid arthritis, be made up of the extract of following weight portion: attached art polysaccharide 15 ~ 35%, atractylodes lactone 23 ~ 43%, Radix Aconiti Lateralis Preparata alkaloid 30 ~ 50%.
Preferably, the ratio of described attached art polysaccharide is 25.5%, and the ratio of atractylodes lactone is 33.0%, and the ratio of Radix Aconiti Lateralis Preparata alkaloid is 41.5%.
Preferably, described attached art polysaccharide is extracted by Radix Aconiti Lateralis Preparata and the Rhizoma Atractylodis Macrocephalae, and described atractylodes lactone is extracted by the Rhizoma Atractylodis Macrocephalae, and described Radix Aconiti Lateralis Preparata alkaloid is extracted by RADIX ACONITI LATERALIS PREPARATA.
In order to realize above-mentioned second object, present invention employs following technical scheme:
Treat a preparation method for the Chinese medicine composition of rheumatoid arthritis, comprise the following steps:
1) preparation of attached art polysaccharide: take Radix Aconiti Lateralis Preparata and Rhizoma Atractylodis Macrocephalae 1000g, wherein Radix Aconiti Lateralis Preparata consumption is 3 times of the Rhizoma Atractylodis Macrocephalae, add 6 times amount volumes and boiling point is the Petroleum ether extraction 2 times of 60 DEG C-90 DEG C, each extraction is after 3 hours, collect medicinal residues and add 80% ethanol, 100 DEG C of water-bath reflux, extract, 2 times, each 3 hours, each 8 times amount solvents, remove fat, pigment, monosaccharide and the oligosaccharide in medical material; After collecting medicinal residues 60 DEG C of vacuum drying 12h, add 10 times of water gagings 100 DEG C and extract 3 hours, extract twice altogether, merge extractive liquid, be evaporated to the medicinal liquid that concentration is 1 gram of crude drug every milliliter, centrifugal removing insoluble matter; Get concentrated medicament, slowly add dehydrated alcohol, limit edged stirs and makes the ultimate density of ethanol in solution be 80%, 4 DEG C of precipitate with ethanol 12h; Sucking filtration obtains light brown crude polysaccharides, and after crude polysaccharides removes protein by SAVEGE method, precipitate with ethanol again, then washs 3 times successively with dehydrated alcohol, acetone, be transferred in evaporating dish, and vacuum drying obtains attached art polysaccharide;
2) preparation of atractylodes lactone: take Rhizoma Atractylodis Macrocephalae 1000g, adds 75% ethanol, 100 DEG C of water-bath reflux, extract, 2 times, each 3 hours, each 8 times amount solvents; Merge extractive liquid, is evaporated to the medicinal liquid that concentration is 1 gram of crude drug every milliliter, and centrifugal removing insoluble matter adds to D-101 macroporous resin column upper end, standing adsorption 1 hours; 6 times of column volume pure water eluting, discard eluent, 5 times of column volume 70% ethanol elutions, and collect eluent, be concentrated into small size, vacuum drying obtains atractylodes lactone;
3) preparation of Radix Aconiti Lateralis Preparata alkaloid: take RADIX ACONITI LATERALIS PREPARATA medical material 1000g, add 75% ethanol, 100 DEG C of water-bath reflux, extract, 2 times, each 3 hours, each 8 times amount solvents; Merge extractive liquid, is evaporated to the medicinal liquid that concentration is 1 gram of crude drug every milliliter, and centrifugal removing insoluble matter adds to D-101 macroporous resin column upper end, standing adsorption 1 hours; 6 times of column volume pure water eluting, discard eluent, 5 times of column volume 70% ethanol elutions, and collect eluent, be concentrated into small size, vacuum drying obtains Radix Aconiti Lateralis Preparata alkaloid.
Preferably, the optimization of described attached art polysaccharide, atractylodes lactone and Radix Aconiti Lateralis Preparata alkaloid three proportion compatibility, adopt simple grid shape to design and carry out component proportion experiment, and use the response surface and crave for function method and carry out multiple-objection optimization, set up active component and the relation between compatibility proportioning mode and drug effect thereof.
Used simple grid shape design in the present invention, 10 groups are combined the ratio of seeking optimum combination altogether, and group is few, credible result.Compared with prior art, the active component of Radix Aconiti Lateralis Preparata of the present invention and Rhizoma Atractylodis Macrocephalae medicine centering carries out reasonable compatibility, it is made to be well used preparing in drugs for rheumatoid arthritis, for finding the thinking and countermeasure that new component compatibility provides new from complicated Chinese medicine system, can be used as the exploitation of Chinese medicine two kind new medicine.
Accompanying drawing explanation
Fig. 1 is the FT-IR spectrogram of attached art polysaccharide component.
Fig. 2 be atractylenolideⅡ (A, 276nm) in atractylodes lactone component, I, the HPLC of III (B, 220nm) figure.
Fig. 3 is the HPLC figure of Radix Aconiti Lateralis Preparata alkaloid active component.
Fig. 4 is that Radix Aconiti Lateralis Preparata Rhizoma Atractylodis Macrocephalae medicine is schemed and contour map the response surface that left sufficient swelling degree of the paw is optimized different prescription.
Fig. 5 is that Radix Aconiti Lateralis Preparata Rhizoma Atractylodis Macrocephalae medicine is schemed and contour map the response surface that IL-1 β in serum optimizes different prescription.
Fig. 6 is that Radix Aconiti Lateralis Preparata Rhizoma Atractylodis Macrocephalae medicine is schemed and contour map the response surface that 5-HT in serum optimizes different prescription.
Fig. 7 is the response value of D value binomial model and the scattergram of predictive value.
Detailed description of the invention
The present invention further describes flesh and blood of the present invention in conjunction with the accompanying drawings and embodiments, and this embodiment is not only limitation of the present invention for illustration of the present invention.
The preparation of one active component
1. the preparation of the attached art polysaccharide of active component
Take Radix Aconiti Lateralis Preparata and Rhizoma Atractylodis Macrocephalae 1000g (Radix Aconiti Lateralis Preparata: the Rhizoma Atractylodis Macrocephalae=3:1), add 6 times amount volume petroleum ether (boiling point: 60-90 DEG C) to extract 2 times, each extraction is after 3 hours, collect medicinal residues and add 80% ethanol, 100 DEG C of water-bath reflux, extract, 2 times, each 3 hours, each 8 times amount solvents, removed the fat in medical material, pigment, monosaccharide and oligosaccharide.After collecting medicinal residues 60 DEG C of vacuum drying 12h, add 10 times of water gagings 100 DEG C and extract 3 hours, extract twice altogether, merge extractive liquid, be evaporated to the medicinal liquid that concentration is 1 gram of crude drug every milliliter, centrifugal removing insoluble matter.Get concentrated medicament, slowly add dehydrated alcohol, limit edged stirs and makes the ultimate density of ethanol in solution be 80%, 4 DEG C of precipitate with ethanol 12h.Sucking filtration obtains light brown crude polysaccharides, and after SAVEGE method removes protein, precipitate with ethanol again, dehydrated alcohol, acetone wash 3 times successively, and be transferred in evaporating dish, vacuum drying is for subsequent use.
2. the preparation of active component atractylodes lactone
Take Rhizoma Atractylodis Macrocephalae 1000g, add 75% ethanol, 100 DEG C of water-bath reflux, extract, 2 times, each 3 hours, each 8 times amount solvents.Merge extractive liquid, is evaporated to the medicinal liquid that concentration is 1 gram of crude drug every milliliter, and centrifugal removing insoluble matter adds to D-101 macroporous resin column upper end, standing adsorption 1 hours.6 times of column volume pure water eluting, discard eluent, 5 times of column volume 70% ethanol elutions, and collect eluent, be concentrated into small size, vacuum drying is for subsequent use.
3. the preparation of active component Radix Aconiti Lateralis Preparata alkaloid
Take RADIX ACONITI LATERALIS PREPARATA medical material 1000g, add 75% ethanol, 100 DEG C of water-bath reflux, extract, 2 times, each 3 hours, each 8 times amount solvents.Merge extractive liquid, is evaporated to the medicinal liquid that concentration is 1 gram of crude drug every milliliter, and centrifugal removing insoluble matter adds to D-101 macroporous resin column upper end, standing adsorption 1 hours.6 times of column volume pure water eluting, discard eluent, 5 times of column volume 70% ethanol elutions, and collect eluent, be concentrated into small size, vacuum drying is for subsequent use.
The analysis of two active components
1. attached art polysaccharide active component is analyzed
Precision takes through 50 DEG C of vacuum drying polysaccharide 5mg, is dissolved in water, is settled in 50mL volumetric flask.The solution of joining be 0.1gmL -1need testing solution.Pipette 0.1mgmL respectively -1dextrose standard sample solution 0.2,0.4,0.6,0.8,1.0,1.2mL in dry tool plug test tube, adding distil water is supplemented to 2mL, using 2mL distilled water as blank, measures absorbance A value by phend-sulphuric acid at 489nm place.And get the polysaccharide sample solution 1mL prepared and be placed in dry tool plug test tube, working sample absorbance as stated above.The content calculating wherein total sugar according to standard curve and sample concentration is 65.16%.
Separately get appropriate attached art polysaccharide sample, adopt KBr pressed disc method, at 4000-400cm -1carry out IR spectrum scanning in scope, scanning times is 32 times, and resolution is 4cm -1, the results are shown in Figure 1.From in figure, polysaccharide sample is at 2929.42cm -1the absworption peak at place is the stretching vibration absorption of C-H, 1418.94cm -1the absworption peak at place is the bending vibration absorption of C-H, and these two groups of absworption peaks are characteristic absorption peaks of saccharide compound; 3421.95cm -1the strong absworption peak at place is the stretching vibration absorption of O-H key; 1644.93cm -1the absworption peak at place is the absorption of Bound moisture; 1155.78,1080.65 and 1022.42cm -1place absworption peak be in pyranose ring structure C=O wash receipts peak, pyranoside has 3 absworption peaks at 1100-1010 place; 844.05cm -1the absworption peak at place is the angle vibration of α-D-pyranose C-H, and show that this polysaccharide component contains α-D-pyranose glucose ring, molecule connects with α-glycosidic bond.
2. atractylodes lactone active component is analyzed
Separately get homemade atractylodes lactone component 0.2632g, dissolve with methanol, after ultrasonic 30min, centrifugal, get supernatant and be settled to 10mL, cross the organic membrane of 0.22 μm, obtain need testing solution for subsequent use.
HPLC condition: chromatographic column is C 18(4.6mm × 250mm, 5 μm); Mobile phase A: acetonitrile, Mobile phase B: water, gradient elution program is in table 1; Flow velocity: 1mLmin -1; Column temperature: 30 DEG C; Sample size: 10 μ L.
The condition of gradient elution of table 1 atractylodes lactone component
Sequence Time (min) A％ B％
1 0 60 40
2 16 70 30
3 18 65 35
6 30 65 35
The efficient liquid phase chromatographic analysis spectrogram of atractylodes lactone active component is shown in Fig. 2.HPLC measure atractylenolide Ⅰ in atractylodes lactone component, II, the content of III is respectively 202.14mgg -1, 86.40mgg -1, 321.30mgg -1.
3. Radix Aconiti Lateralis Preparata alkaloid active component is analyzed
Take self-control alkaloidal constituent powder 0.1000g, add ethanol 5ml and dissolve, adjust PH to 10-11, chloroform extraction 2 times, each 10ml, gets chloroform layer and volatilizes, and residue adds ethanol 5mL makes dissolving, and precision adds sulphuric acid volumetric solution (0.01molL -1) 15mL, water 15mL and C.I. 13020. indicator solution 3, with sodium hydroxide titration liquid (0.02molL -1) be titrated to yellow.With 1mL sulphuric acid volumetric solution (0.01molL -1) be equivalent to the Aconitum carmichjaelii Debx. (C of 12.9mg 34h 47nO 11) calculate, in 0.1g alkaloidal constituent, the content of total alkaloids is 52.096mg.
Precision takes homemade alkaloid active component 0.1008g, is dissolved in 10mL methanol, ultrasonic 30min, centrifugal, gets supernatant, adds methanol constant volume to 10mL, crosses the organic membrane of 0.22 μm, obtains need testing solution for subsequent use.
HPLC condition: chromatographic column is Dalian Yi Lite BDSC 18post (4.6 × 250mm, 5 μm); Mobile phase: methanol: 0.15% triethylamine-aqueous solution gradient elution, gradient condition in table 2, flow velocity 1mLmin -1.
The elution requirement of table 2 alkaloidal constituent
Time Methanol 0.15% triethylamine-aqueous solution
0 55 45
7 56 44
10 57 43
15 60 40
30 69 31
35 73 27
40 75 25
The efficient liquid phase chromatographic analysis spectrogram of Radix Aconiti Lateralis Preparata alkaloid active component is shown in Fig. 3, and HPLC measures alkaloidal constituent mesaconitine, mesaconitine and hypaconitine content and is respectively 72.10mgg -1, 96.24mgg -1, 367.76mgg -1.
The effect experiment of three different active component compatibility proportionings
This experiment adopts simple grid shape experimental design, select 3 kinds of components of Radix Aconiti Lateralis Preparata Rhizoma Atractylodis Macrocephalae medicine centering, Radix Aconiti Lateralis Preparata Rhizoma Atractylodis polysaccharide (A), atractylodes lactone (B) and Radix Aconiti Lateralis Preparata total alkaloids (C) form 3 factors, (300mgkgd under same dosage -1), have employed that { 3,3} simplex lattice design, testing site number is C 3 3 + 3-1=10.In order to simplify calculating, the concentration of each component has been carried out standardization, and Cmin is classified as 0, and maximum concentration is classified as 1.Each point Xi value is 0,1/3,2/3 and 1.Planning of experiment is as shown in table 3.Wherein in Radix Aconiti Lateralis Preparata total alkaloids component because of containing toxic stronger diester-type alkaloids, according to the limited amount regulation of your diester-type alkaloids, the maximal dose of the diester-type alkaloids related in literary composition is all lower than the LD of rat in preliminary experiment 50.
The simple grid shape experimental design table of table 3
Group Active component A Active component B Active component C Drug effect Y 4-->
C1 1 0 0 Y1
C2 0 1 0 Y2
C3 0 0 1 Y3
C4 2/3 1/3 0 Y4
C5 2/3 0 1/3 Y5
C6 1/3 0 2/3 Y6
C7 1/3 2/3 0 Y7
C8 0 1/3 2/3 Y8
C9 0 2/3 1/3 Y9
C10 1/3 1/3 1/3 Y10
After Rat Right metapedes routine disinfection, inject Freund's complete adjuvant 0.02mL to sufficient sole of the foot pad portion to ankle joint direction and make adjuvant-induced arthritis animal model.Select modeling success SD rat, second day after modeling starts administration, according to simple grid shape design, three combination of components is become 10 prescriptions, is divided into into 10 groups, often organizes 8 rats, every day 300mgkg -1the heavy gastric infusion of Mus, arranges model group and normal group to give equal-volume normal saline in addition.Successive administration by sacrifice of animal, got blood 3mL after 21 days, put in Eppendorf pipe, and the centrifugal 15min of 3500rpm, gets serum for subsequent use, and ELISA method measures IL-1 β content in serum; Peel off full brain simultaneously, get 1/2 fresh brain tissue, after cleaning with normal saline, dry, weigh, homogenate, the centrifugal 15min of 3000rpm, gets supernatant, and ELISA method measures the content of 5-HT in maincenter cerebral tissue.Measure left whole sole of the foot thickness with thickness gauge and calculate swelling degree of the paw.
Table 4 is the impact on the left sufficient swelling degree of the paw of adjuvant arthritis rats, IL-1 β and 5-HT after the Radix Aconiti Lateralis Preparata Rhizoma Atractylodis Macrocephalae and different prescription administration.As can be seen from table, after rat modeling, swelling degree of the paw and IL-1 β content all have rising (P<0.05 largely, P<0.05), and cental system 5-HT content slightly increase (P>0.05) because of pain stimulation after modeling.Attached art polysaccharide, atractylodes lactone and the single component of Radix Aconiti Lateralis Preparata alkaloid on swelling degree of the paw, IL-1 β and 5-HT all has affects extremely significantly, P value is all less than 0.01.And all prescriptions all can reduce the content of IL-1 β in the swelling degree of the paw of rat and serum extremely significantly, raise the content of 5-HT in cerebral tissue, P value is all less than 0.01.
The different prescription of the table 4 Radix Aconiti Lateralis Preparata Rhizoma Atractylodis Macrocephalae on the impact of swelling degree of the paw, IL-1 β and 5-HT ( n=8)
Note: compare with blank group ▲p<0.05, ▲ ▲p<0.01; Compare with model group *p<0.05, *p<0.01.
The optimization of four active component compatibility proportionings
Although had a lot of method at present if evolution and genetic algorithm are for multiple-objection optimization, consider the response surface and craved for the unique advantage of function on optimizing, therefore adopted Response Surface Method and crave for function to complete compatibility optimization.Wherein response phase method adopts DesignExpert8.0.6 software to carry out, by the relation between binary polynomial equation reflection variable and response value; And crave for function and can crave for value thus the optimal value reaching expectation by multiple response variable being changed into, here, we are first by each desired value Y ivalue changes into respectively independently craves for value d, craves for value between 0 ~ 1, if when response value is outside acceptable scope, if d i=0, if its response value is considered to perfect, if d i=1.If what response value was pursued is be the bigger the better, then concrete computing formula is as follows:
d i = 0 Y i &le; Y i - m i n &lsqb; Y i - Y i - m i n Y i - max - Y i - m i n &rsqb; r Y i - min < Y i < Y i - max 1 Y i &GreaterEqual; Y i - max
If the response value pursued is the smaller the better, then concrete computing formula is as follows:
d i = 1 Y i &le; Y i - m i n &lsqb; Y i - Y i - m a x Y i - m i n - z i - m a x &rsqb; r Y i - min < Y i < Y i - max 0 Y i &GreaterEqual; Y i - max
Wherein Y i-minfor acceptable minimum Y idesired value, and Y i-maxbe then maximum desired value, r is normal number.Finally independently crave for value in conjunction with all, then crave for function D value to realize total optimization by formulae discovery entirety below:
D = ( d 1 w 1 d 2 w 2 d 3 w 3 d 4 w 4 d n w n ) 1 / &Sigma;w i
Wherein D represents that entirety craves for value, d irepresent and independently crave for value, W irepresent the weight coefficient of i-th d value.
According to variance analysis, draw the multivariate regression equation of swelling degree of the paw, IL-1 β and 5-HT, represent with (1), (2) and (3) respectively.
Y 1＝31.05A+28.78B+31.16C-7.15AB-10.59AC-24.99BC(1)
Y 2＝53.39A+53.35B+47.46C-36.28AB-38.34AC-30.29BC(2)
Y 3＝662.38A+645.44B+708.42C+924.58AB+1162.29AC+1448.59BC(3)
Can find out that from the equal pitch contour and response surface figure of Fig. 4-Fig. 6 three components share the content of the IL-1 β that can significantly reduce in swelling degree of the paw and serum, increase the content of 5-HT in cerebral tissue.Wherein, the effectiveness comparison of atractylodes lactone component and Radix Aconiti Lateralis Preparata alkaloid compatibility is remarkable.Three Y ivalue (Y 1, Y 2, Y 3) change into corresponding d respectively ivalue (d 1, d 2, d 3), in order to reach the anti-inflammatory pain-stopping effect of Radix Aconiti Lateralis Preparata Rhizoma Atractylodis Macrocephalae medicine centering three component the bests, the left sufficient swollen tissue degree (d of its rat 1) value the smaller the better, serum IL-1 β (d 2) value be the bigger the better, the value (d of 5-HT in cerebral tissue 3) be the bigger the better.Therefore, by left for rat sufficient swollen tissue degree (d 1) minima be set to 1.08%, maximum is then set to 100%; Serum IL-1 β (d 2) theoretical minimum value be 0, maximum is then set to 100pgml -1; 5-HT (d in cerebral tissue 3) the theoretical minimum value of content be 0, maximum is set to 1500ngml -1.Therefore, the value of d1, d2, d3 is respectively calculated as follows:
d 1 = 1 Y i &le; 1.08 &lsqb; Y i - 100 1.08 - 100 &rsqb; 1.08 < Y i < 100 0 Y i &GreaterEqual; 100
d 2 = 1 Y i &le; 0 &lsqb; Y i - 100 0 - 100 &rsqb; 0 < Y i < 100 0 Y i &GreaterEqual; 100
d 3 = 0 Y i &le; 0 &lsqb; Y i - 0 1500 - 0 &rsqb; 0 < Y i < 100 1 Y i &GreaterEqual; 100
Therefore, entirety craves for being calculated as follows of value D:
D＝(d 1d 2d 3) 1/3
Each index of final gained independently crave for value d 1, d 2, d 3and entirety crave for value D specifically in table 5, use Response Surface Method D value is optimized, obtaining regression equation is:
D＝0.52A+0.52B+0.56C+0.39AB+0.45AC+0.54BC+0.16ABC
The different prescription of the table 5 Radix Aconiti Lateralis Preparata Rhizoma Atractylodis Macrocephalae crave for functional value
Fig. 7 is the response value of D value binomial model and the distribution of predictive value, and upper as can be seen from figure, most point all drops near diagonal, and the deviation between response value and predictive value is little, shows that models fitting data are good.Application response surface optimization, the optimum combination finally filtered out is Radix Aconiti Lateralis Preparata Rhizoma Atractylodis polysaccharide (A), atractylodes lactone (B) and Radix Aconiti Lateralis Preparata alkaloid (C) account for 25.5%, 33.0% and 41.5%, i.e. 51:66:83 respectively.
The checking of five optimization prescriptions
In order to verification computation result, carry out experimental verification with identical animal model.By optimize the best compatibility side that obtains respectively with atractylodes lactone+Radix Aconiti Lateralis Preparata alkaloid (V1 group, 1:1), blank group, model group, Radix Aconiti Lateralis Preparata Rhizoma Atractylodis Macrocephalae medicine compare research to compatibility group.As shown in table 6, compared with model group, best compatibility obviously can reduce swelling degree of the paw and serum IL-1 β content (P<0.01, P<0.01), and can increase the content (P<0.01) of 5-HT in brain; The simultaneously best compatibility side effect that reduces swelling degree of the paw and serum IL-1 β content and raise 5-HT content in brain and Radix Aconiti Lateralis Preparata Rhizoma Atractylodis Macrocephalae medicine are to compared with compatibility group, except swelling degree of the paw is without except significant statistical significance (P<0.05), namely without outside significant difference, all the other two indices are all better than Radix Aconiti Lateralis Preparata Rhizoma Atractylodis Macrocephalae medicine to compatibility group, there is significant statistical significance (P<0.05, P<0.05); Namely the anti-inflammatory pain-stopping effect of best compatibility side is better than Radix Aconiti Lateralis Preparata Rhizoma Atractylodis Macrocephalae medicine to compatibility.
Table 6 Radix Aconiti Lateralis Preparata Rhizoma Atractylodis Macrocephalae medicine on component compatibility optimization side on the impact of swelling degree of the paw, IL-1 β and 5-HT ( n=8)
Note: compare with blank group ▲p<0.05, ▲ ▲p<0.01; Compare with model group *p<0.05, *p<0.01; With Radix Aconiti Lateralis Preparata Rhizoma Atractylodis Macrocephalae medicine, compatibility group is compared #p<0.05, ##p<0.01.
Used simple grid shape design in the present invention, 10 groups are combined the ratio of seeking optimum combination altogether, and group is few, credible result.Compared with prior art, the active component of Radix Aconiti Lateralis Preparata of the present invention and Rhizoma Atractylodis Macrocephalae medicine centering carries out reasonable compatibility, it is made to be well used preparing in drugs for rheumatoid arthritis, for finding the thinking and countermeasure that new component compatibility provides new from complicated Chinese medicine system, can be used as the exploitation of Chinese medicine two kind new medicine.

Claims (5)

1. treat a Chinese medicine composition for rheumatoid arthritis, it is characterized in that, be made up of the extract of following weight portion: attached art polysaccharide 15 ~ 35%, atractylodes lactone 23 ~ 43%, Radix Aconiti Lateralis Preparata alkaloid 30 ~ 50%.
2. a kind of Chinese medicine composition for the treatment of rheumatoid arthritis according to claim 1, is characterized in that, the ratio of described attached art polysaccharide is 25.5%, and the ratio of atractylodes lactone is 33.0%, and the ratio of Radix Aconiti Lateralis Preparata alkaloid is 41.5%.
3. a kind of Chinese medicine composition for the treatment of rheumatoid arthritis according to claim 1 and 2, is characterized in that, described attached art polysaccharide is extracted by Radix Aconiti Lateralis Preparata and the Rhizoma Atractylodis Macrocephalae, and described atractylodes lactone is extracted by the Rhizoma Atractylodis Macrocephalae, and described Radix Aconiti Lateralis Preparata alkaloid is extracted by RADIX ACONITI LATERALIS PREPARATA.
4. treat a preparation method for the Chinese medicine composition of rheumatoid arthritis, it is characterized in that, comprise the following steps:
1) preparation of attached art polysaccharide: take Radix Aconiti Lateralis Preparata and Rhizoma Atractylodis Macrocephalae 1000g, wherein Radix Aconiti Lateralis Preparata consumption is 3 times of the Rhizoma Atractylodis Macrocephalae, add 6 times amount volumes and boiling point is the Petroleum ether extraction 2 times of 60 DEG C-90 DEG C, each extraction is after 3 hours, collect medicinal residues and add 80% ethanol, 100 DEG C of water-bath reflux, extract, 2 times, each 3 hours, each 8 times amount solvents, remove fat, pigment, monosaccharide and the oligosaccharide in medical material; After collecting medicinal residues 60 DEG C of vacuum drying 12h, add 10 times of water gagings 100 DEG C and extract 3 hours, extract twice altogether, merge extractive liquid, be evaporated to the medicinal liquid that concentration is 1 gram of crude drug every milliliter, centrifugal removing insoluble matter; Get concentrated medicament, slowly add dehydrated alcohol, limit edged stirs and makes the ultimate density of ethanol in solution be 80%, 4 DEG C of precipitate with ethanol 12h; Sucking filtration obtains light brown crude polysaccharides, and after crude polysaccharides removes protein by SAVEGE method, precipitate with ethanol again, then washs 3 times successively with dehydrated alcohol, acetone, be transferred in evaporating dish, and vacuum drying obtains attached art polysaccharide;
2) preparation of atractylodes lactone: take Rhizoma Atractylodis Macrocephalae 1000g, adds 75% ethanol, 100 DEG C of water-bath reflux, extract, 2 times, each 3 hours, each 8 times amount solvents; Merge extractive liquid, is evaporated to the medicinal liquid that concentration is 1 gram of crude drug every milliliter, and centrifugal removing insoluble matter adds to D-101 macroporous resin column upper end, standing adsorption 1 hours; 6 times of column volume pure water eluting, discard eluent, 5 times of column volume 70% ethanol elutions, and collect eluent, be concentrated into small size, vacuum drying obtains atractylodes lactone;
3) preparation of Radix Aconiti Lateralis Preparata alkaloid: take RADIX ACONITI LATERALIS PREPARATA medical material 1000g, add 75% ethanol, 100 DEG C of water-bath reflux, extract, 2 times, each 3 hours, each 8 times amount solvents; Merge extractive liquid, is evaporated to the medicinal liquid that concentration is 1 gram of crude drug every milliliter, and centrifugal removing insoluble matter adds to D-101 macroporous resin column upper end, standing adsorption 1 hours; 6 times of column volume pure water eluting, discard eluent, 5 times of column volume 70% ethanol elutions, and collect eluent, be concentrated into small size, vacuum drying obtains Radix Aconiti Lateralis Preparata alkaloid.
5. a kind of preparation method for the treatment of the Chinese medicine composition of rheumatoid arthritis according to claim 1, it is characterized in that, the optimization of described attached art polysaccharide, atractylodes lactone and Radix Aconiti Lateralis Preparata alkaloid three proportion compatibility, adopt simple grid shape to design and carry out component proportion experiment, and use the response surface and crave for function method and carry out multiple-objection optimization, set up active component and the relation between compatibility proportioning mode and drug effect thereof.
CN201511019368.3A 2015-12-30 2015-12-30 A kind of Chinese medicine composition and preparation method thereof for treating rheumatoid arthritis Active CN105434567B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201511019368.3A CN105434567B (en) 2015-12-30 2015-12-30 A kind of Chinese medicine composition and preparation method thereof for treating rheumatoid arthritis

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201511019368.3A CN105434567B (en) 2015-12-30 2015-12-30 A kind of Chinese medicine composition and preparation method thereof for treating rheumatoid arthritis

Publications (2)

Publication Number Publication Date
CN105434567A true CN105434567A (en) 2016-03-30
CN105434567B CN105434567B (en) 2019-09-10

Family

ID=55545501

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201511019368.3A Active CN105434567B (en) 2015-12-30 2015-12-30 A kind of Chinese medicine composition and preparation method thereof for treating rheumatoid arthritis

Country Status (1)

Country Link
CN (1) CN105434567B (en)

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102166302A (en) * 2011-03-31 2011-08-31 王一军 Metabolism-compensation method for treating rheumatic osteoarthritis and proportioning and preparing process of preparation thereof
CN102552223A (en) * 2012-01-17 2012-07-11 浙江省中医药研究院 Application of alpha-terpinenol to treatment of arthritis
CN102895617A (en) * 2011-07-29 2013-01-30 苏州知微堂生物科技有限公司 Preparation technology and production method for integrated new formulation of bighead atractylodes rhizome and monkshood decoction
CN104688846A (en) * 2013-12-04 2015-06-10 王晓娟 acute rheumatoid arthritis
CN104784515A (en) * 2015-04-08 2015-07-22 宋长银 Traditional Chinese medicine for treating rheumatic arthritis

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102166302A (en) * 2011-03-31 2011-08-31 王一军 Metabolism-compensation method for treating rheumatic osteoarthritis and proportioning and preparing process of preparation thereof
CN102895617A (en) * 2011-07-29 2013-01-30 苏州知微堂生物科技有限公司 Preparation technology and production method for integrated new formulation of bighead atractylodes rhizome and monkshood decoction
CN102552223A (en) * 2012-01-17 2012-07-11 浙江省中医药研究院 Application of alpha-terpinenol to treatment of arthritis
CN104688846A (en) * 2013-12-04 2015-06-10 王晓娟 acute rheumatoid arthritis
CN104784515A (en) * 2015-04-08 2015-07-22 宋长银 Traditional Chinese medicine for treating rheumatic arthritis

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
张锟等: "中医药治疗类风湿关节炎研究进展", 《风湿病与关节炎》 *
范先志: "桂芍知母汤联合来氟米特对急性风湿关节炎的疗效与安全性分析", 《辽宁中医杂志》 *
陈佩虹等: "寒湿痹片抑制大鼠滑膜增生及相关机制研究", 《中国实验方剂学杂志》 *

Also Published As

Publication number Publication date
CN105434567B (en) 2019-09-10

Similar Documents

Publication Publication Date Title
Kuang et al. Screening and comparison of the immunosuppressive activities of polysaccharides from the stems of Ephedra sinica Stapf
CN102631390B (en) Preparation method of periploca forrestii schltr extract as well as product and application of periploca forrestii schltr extract
CN105169096B (en) A kind of pharmaceutical composition for treating gout or/and hyperuricemia
Xu et al. Recognition and optimization of ingredients treating nitroglycerin-induced migraine rats from Wuzhuyu decoction
CN103191289B (en) Synchronous preparation method of four effective parts in medicine pair of common anemarrhena rhizome and amur corktree bark and application thereof
CN101810715B (en) New application of fructus evodiae and extracts and compounds thereof
CN103463156B (en) A kind of Heiguteng exract extract and its production and use
CN102100825B (en) Chinese medicine composition for treating pains and preparation method thereof
WO2022022648A1 (en) Traditional chinese medicine composition having mental relief and antidepressant effects and preparation method therefor
CN101269179B (en) Traditional Chinese medicine preparation for treating senile dementia and preparation method thereof
CN100446805C (en) Medicine for treating chronic diarrhoea
CN102133242B (en) Tuckahoe extract and preparation method thereof
CN101574377B (en) Effective part of sweetberry jointfir, preparation method and application thereof in preparing medicament for treating gout disease
CN105434567A (en) Traditional Chinese medicine composition for treating rheumatoid arthritis and preparation method thereof
CN101642492B (en) Drug for treating chronic pelvic inflammatory disease and preparation method thereof
CN103156997B (en) Composition of effective parts of traditional Chinese medicines for treating chronic hepatopathy, preparation method and application thereof
CN103837636A (en) HPTLC (high performance thin layer chromatography) quality evaluation method of Xinfeng capsule by adopting coix seeds as reference
CN109270203B (en) Construction method of characteristic spectrum of active ingredients of Jingyaokang capsule and quality detection method of Jingyaokang capsule
CN1434042A (en) Cattail pollen extract and preparation process and use thereof
CN104547514A (en) Traditional Chinese medicine composition for treating systemic lupus erythematosus rheumatoid arthritis vasculitis and application thereof
CN105616556B (en) A kind of preparation method preventing and treating alcoholic liver injury active principle
CN104069204A (en) Scutellaria baicalensis decoction granules for treating ulcerative colitis and preparation method thereof
CN104161859A (en) Traditional Chinese medicine composition for treating spleen and stomach diseases, and preparation method thereof
CN107998316A (en) It is a kind of to be used to treat Chinese medicine composition of allergic dermatitis and preparation method thereof
CN103301414B (en) Gout, short kidney are arranged, repair kidney damages pharmaceutical composition and its preparation method and application

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant