CN104274540B - Chinese medicine composition, preparation method and use containing tripterygium wilfordii - Google Patents
Chinese medicine composition, preparation method and use containing tripterygium wilfordii Download PDFInfo
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- WVYADZUPLLSGPU-UHFFFAOYSA-N salsalate Chemical compound OC(=O)C1=CC=CC=C1OC(=O)C1=CC=CC=C1O WVYADZUPLLSGPU-UHFFFAOYSA-N 0.000 description 1
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 1
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 1
- 229960004025 sodium salicylate Drugs 0.000 description 1
- JGMJQSFLQWGYMQ-UHFFFAOYSA-M sodium;2,6-dichloro-n-phenylaniline;acetate Chemical compound [Na+].CC([O-])=O.ClC1=CC=CC(Cl)=C1NC1=CC=CC=C1 JGMJQSFLQWGYMQ-UHFFFAOYSA-M 0.000 description 1
- 238000001256 steam distillation Methods 0.000 description 1
- 230000003637 steroidlike Effects 0.000 description 1
- NCEXYHBECQHGNR-QZQOTICOSA-N sulfasalazine Chemical compound C1=C(O)C(C(=O)O)=CC(\N=N\C=2C=CC(=CC=2)S(=O)(=O)NC=2N=CC=CC=2)=C1 NCEXYHBECQHGNR-QZQOTICOSA-N 0.000 description 1
- 229960001940 sulfasalazine Drugs 0.000 description 1
- 229960000894 sulindac Drugs 0.000 description 1
- MLKXDPUZXIRXEP-MFOYZWKCSA-N sulindac Chemical compound CC1=C(CC(O)=O)C2=CC(F)=CC=C2\C1=C/C1=CC=C(S(C)=O)C=C1 MLKXDPUZXIRXEP-MFOYZWKCSA-N 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 210000002435 tendon Anatomy 0.000 description 1
- 230000008719 thickening Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 210000003371 toe Anatomy 0.000 description 1
- 229960001017 tolmetin Drugs 0.000 description 1
- UPSPUYADGBWSHF-UHFFFAOYSA-N tolmetin Chemical compound C1=CC(C)=CC=C1C(=O)C1=CC=C(CC(O)=O)N1C UPSPUYADGBWSHF-UHFFFAOYSA-N 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000820 toxicity test Toxicity 0.000 description 1
- 231100000925 very toxic Toxicity 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- 210000002268 wool Anatomy 0.000 description 1
- 230000037314 wound repair Effects 0.000 description 1
- 239000008096 xylene Substances 0.000 description 1
Classifications
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- 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)
- A61K36/37—Celastraceae (Staff-tree or Bittersweet family), e.g. tripterygium or spindletree
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
- A61K36/232—Angelica
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- A61K36/185—Magnoliopsida (dicotyledons)
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- A61K36/236—Ligusticum (licorice-root)
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/32—Burseraceae (Frankincense family)
- A61K36/324—Boswellia, e.g. frankincense
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- 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)
- A61K36/32—Burseraceae (Frankincense family)
- A61K36/328—Commiphora, e.g. mecca myrrh or balm of Gilead
-
- 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)
- A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
- A61K36/534—Mentha (mint)
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/70—Polygonaceae (Buckwheat family), e.g. spineflower or dock
- A61K36/708—Rheum (rhubarb)
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- A61K2236/30—Extraction of the material
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Abstract
Tripterygium wilfordii, saltcake, Rhizoma Chuanxiong, the root of Dahurain angelica, rhubarb, frankincense（System）, myrrh（System）, peppermint Chinese medicine composition, available for treatment rheumatoid arthritis or prepare treatment rheumatoid arthritis drug.
Description
Technical field
The present invention relates to a kind of Chinese medicine composition containing tripterygium wilfordii, available for the medicine for preparing treatment rheumatoid arthritis
Object.The present invention also relates to the preparation methods and purposes of the Chinese medicine composition containing tripterygium wilfordii.
Background technology
Rheumatoid arthritis（Rheumatoid Arthritis, RA）It is slow as what is mainly showed using symmetry panarthritis
Property, progressive, aggressivity disease.RA is one kind using synovium of joint chronic inflammation as the autoimmune disease mainly showed.Mesh
Before, the illness rate in the world is 1%, and the illness rate in China is 0.3-0.4%, and there are about 4,000,000 patients in continent, it is seen that harm face is suitable
Extensively.Beijing area be this sick district occurred frequently, illness rate 0.34%.This disease can cause multiple joints of whole body swelling pain, and late period can
Joint deformity is caused, function is lost, and at least 50% cannot adhere to work in patient 10 years diseased.The disease is summarised as by foreign scholar
5D, i.e., it is painful（discomfort）, it is dead（death）, it is disabled（disability）, economic loss（dollar lost）, drug
Poisoning（drug toxicity）.
The primary therapeutic purpose of RA is by the symptom that controls inflammation, prevents destruction of joint, holding function of joint, maintenance normal
Social activities utmostly improves the long-term health of patient, quality of life.Diminish inflammation is to realize the most important side of this target
Method, clinically the most commonly used is the nonsteroidal anti-inflammatory agents of first-line treatment at present（Non-Steroid Anti-Inflammatory
Drugs, NSAIDs）The glucocorticoid of " bridge " treatment, with methotrexate (MTX)（MTX）For choice drug generally acknowledged both at home and abroad, but
Be since NSAIDs and hormone side effect are more, NSAIDs gastrointestinal tracts, angiocarpy as known to being numerous health care workers,
The osteoporosis of the adverse reactions such as kidney and glucocorticoid, hypertension, diabetes, infection risk etc., prevent many patients from
Using or tolerance, cannot still meet the needs of clinical treatment.
Chinese herbal medicine tripterygium wilfordii is used for antirheumatic (DMARD), recognized by medical field.It is widely used in RA in the past 30 years
Treatment, the case of hitherto reported is efficient up to 80%~90% more than 8,000.Clinical research proves that tripterygium wilfordii is treatment RA
Effective Chinese medicine, and its effective monomer component structure is also it has been determined that this point is approved by international counterparts.But because its is very toxic,
Bone marrow suppression, hepatorenal damage and female patient can be caused there are the adverse reactions such as amenorrhoea, pigmentation, its clinical practice is made to be subject to one
Determine the limitation of degree, it is difficult to adhere to medication for a long time, clearly limitation uses especially in young patient.
The effect of how utilizing tripterygium wilfordii, while various means is taken to lower its toxicity, it is made to play maximum effect, is mesh
The direction that preceding clinician makes great efforts.Clinically need the drug of effectively treatment rheumatoid arthritis.
The content of the invention
In view of it is above-mentioned, applicant carried out research extensively and profoundly, it turns out that, tripterygium wilfordii, saltcake, Rhizoma Chuanxiong, the root of Dahurain angelica, life
Rheum officinale, frankincense（System）, myrrh（System）, peppermint Chinese medicine composition have treatment rheumatoid arthritis activity.
It is an object of the present invention to provide a kind of Chinese medicine compositions for treating rheumatoid arthritis；The present invention another
Purpose is to provide a kind of method for preparing Chinese medicine composition of the present invention.It is a further object to provide one kind to contain this hair
The drug of bright Chinese medicine composition and/or its preparation.It is a further object to provide a kind of Traditional Chinese drug mixture of the present invention or
The purposes of Chinese medical extract, for treating rheumatoid arthritis or being used to prepare the drug for the treatment of rheumatoid arthritis.
Specific embodiment
The amount of Chinese medicine composition each component described in embodiment of the present invention and embodiment represents with weight " gram ", i.e. table
Show a parts by weight.The amount of each component can be identical ratio enlargement several times.
In one embodiment, Chinese medicine composition of the present invention is provided, it is characterised in that tripterygium wilfordii, saltcake, Rhizoma Chuanxiong, white
The root of Dahurian angelica, rhubarb, frankincense（System）, myrrh（System）, peppermint combination.
We are using tripterygium wilfordii as monarch drug in a prescription.Tripterygium wilfordii is bitter, cold, toxic.Converge to heart and liver channels.There is removing damp-heat, it is promoting blood circulation and removing obstruction in channels, disappear
The effect of swollen analgesic is the treatment common effective drug of rheumatoid arthritis.《Chinese medical herb will》Cloud：" tripterygium wilfordii hardship,
It is puckery, it trembles with fear.It is toxic.Function relaxing tendons and activating collaterals, dispelling wind and eliminating dampness.Cure mainly rheumatoid arthritis.”
Saltcake, the ministerial drug that Rhizoma Chuanxiong is we.Saltcake is pungent, salty, bitter, trembles with fear.This product, which has taken orally to purge heat, leads stagnant, the work of moistening for dryness and softening hard mass
With local topical has clearing heat-fire, the effect of swelling and pain relieving.《Haigoushen》Cloud：This product " stop up when main, can dispelling malignant blood by disease heat." river
Rhizome of chuanxiong is pungent, warm, and the pungent scattered temperature of effect is logical, and blood-activating and qi-promoting, wind-expelling pain-stopping is clinically used blood-activating drug.《Drugization justice》Call this product：
" its acrid flavour temperature, energy row profit key, makes blood flow gas, is gas medicine in blood.”《All book on Chinese herbal medicine of solar corona》Calling this product can " dispersing blood stasis blood.”
The root of Dahurain angelica, rhubarb, frankincense, myrrh, peppermint are assistant.Root of Dahurain angelica property and flavor of peppery and warm, have relieve heat, dehumidifying, sensible, apocenosis, only
Pain five functional.Rhubarb is bitter, cold.With clearing heat-fire, the function of blood circulation and removing stasis.Rheum officinale external application also can be clearing heat and detoxicating, promoting blood circulation
Dissolving stasis.《Records of Tradition Chinese and Western Medicine in Combination》Call it：" blood system can be entered, break all hemostasis." frankincense works hard, temperature, myrrh is bitter, flat, and two medicines are equal
With promoting blood circulation, the effect of swelling and pain relieving, Chang Xiangxu is used.Such as《Compendium of Materia Medica》Meaning " invigorate blood circulation, and myrrh dissipates blood, all can by frankincense
Analgesic, detumescence, myogenic, therefore two medicines often mutually and and are used ".Peppermint external application plays an important role of cool pain-alleviating, and often coordinates with the root of Dahurain angelica
Using two medicines play the role of resuscitation with aromatics, clearing away heat to and alleviating pain with being that assistant makes altogether.
Tripterygium wilfordii, saltcake, rhubarb, peppermint all medicines share can enhance removing damp-heat in side, subside a swelling dredging collateral the effect of；River
Rhizome of chuanxiong, frankincense, myrrh, root of Dahurain angelica all medicines share can enhance promoting blood circulation, the effect of removing obstruction in channels to relieve pain.All medicines share, play altogether clearing heat and expelling damp,
The effect of promoting blood circulation and stopping pain.
In further embodiment, Chinese medicine composition of the present invention is provided, it is characterised in that 50 to 70 grams of tripterygium wilfordii, awns
55 to 65 grams of nitre, 20 to 40 grams of Rhizoma Chuanxiong, 20 to 40 grams of the root of Dahurain angelica, 10 to 20 grams of rhubarb, frankincense（System）10 to 20 grams, myrrh（System）
10 to 20 grams, 10 grams to 20 grams of peppermint.
In further embodiment, Chinese medicine composition of the present invention is provided, it is characterised in that 60 grams of tripterygium wilfordii, saltcake
60 grams, 30 grams of Rhizoma Chuanxiong, 30 grams of the root of Dahurain angelica, 15 grams of rhubarb, frankincense（System）15 grams, myrrh（System）15 grams, 15 grams of peppermint.
In another embodiment, the method for preparing Chinese medicine composition of the present invention is provided, it is characterised in that：
(1) each Chinese medicine material is directly mixed, obtains Traditional Chinese drug mixture；Optionally and
(2) mixture obtained by water and/or alcohol extracting above-mentioned (1), obtains Chinese medical extract.
The Chinese medical extract of the present invention can also be prepared according to other methods, such as be prepared by following method：
I) tripterygium wilfordii, rheum officinale, the root of Dahurain angelica are extracted with ethyl alcohol, and filtration, filtrate recycling ethanol is spare,
The another device of the dregs of a decoction preserves；
Ii) Rhizoma Chuanxiong, frankincense, three taste extracting in water volatile oil of myrrh, collect volatile oil, and liquid filtration is spare；
Iii) Rhizoma Chuanxiong, frankincense, the dregs of a decoction of three taste of myrrh merge tripterygium wilfordii, rheum officinale, the dregs of a decoction of three taste of the root of Dahurain angelica, add in saltcake,
It is decocted, filtered with water, filtrate merges other above-mentioned liquids, concentrates, and filtration is spare；
Iv) take more than Rhizoma Chuanxiong, frankincense, three taste medicine of myrrh volatile oil, from peppermint extract peppermint oil dementholized dissolved with ethyl alcohol,
Ethanol is mixed with the liquid of above-mentioned concentration.
In further embodiment, provide Traditional Chinese drug mixture prepared by the method for the present invention or Chinese medical extract or
Contain their Traditional Chinese drug mixture or Chinese medical extract.
In another embodiment, the purposes of Traditional Chinese drug mixture or Chinese medical extract of the present invention is provided, for treating
Rheumatoid arthritis or the drug for being used to prepare treatment rheumatoid arthritis, preferably external used medicine.
In further embodiment, the drug for the treatment of rheumatoid arthritis is provided, with Traditional Chinese drug mixture of the present invention or
Chinese medicine composition is as active ingredient.
In further embodiment, the combination drug for the treatment of rheumatoid arthritis is provided, including in the present invention
Medicine mixture or Chinese medical extract are as effective component of chinese medicine and NSAIDs and/or DMARDs class Western medicine active ingredients.
In another embodiment, the pharmaceutical composition for the treatment of rheumatoid arthritis is provided, including in the present invention
Medicine mixture or Chinese medical extract are as active ingredient, optional Western medicine active ingredient and pharmaceutically useful carrier, diluent or tax
Type agent.
In another embodiment, the pharmaceutical preparation for the treatment of rheumatoid arthritis is provided, includes the medicine of the present invention
Compositions.
In further embodiment, pharmaceutical preparation of the invention is solution, gelling agent, lotion, liniment, coating
Agent, spray, creme, paste or patch, preferably ointment or gelling agent.
In another embodiment, the preparation method for the pharmaceutical preparation for preparing treatment rheumatoid arthritis is provided,
Traditional Chinese drug mixture or Chinese medical extract including the present invention are mixed with pharmaceutically acceptable matrix, are optionally joined with Western medicine of the present invention
With.
In one embodiment, Chinese medicine composition of the present invention can also include other traditional Chinese medicine ingredients, such as Radix Glycyrrhizae etc..
In another embodiment, Traditional Chinese drug mixture of the invention or Chinese medical extract can also be with the west for the treatment of rheumatoid arthritis
Medicine Western medicine such as non-steroid anti-inflammatory drug NSAIDs and/or DMARDs is combined to treat rheumatoid arthritis.
The Western medicine for the treatment of rheumatoid arthritis is well known to those skilled in the art, and representative example resists including non-steroidal
Scorching anodyne such as acetylsalicylic acid salt include aspirin, non-acetylsalicyclic Barbiturates include magnesium salicylate, sodium salicylate,
Choline Salicylate magnesium, Diflunisal（Diflunisal）, sasapyrin, it is beautiful that non-salicylic acid salt includes brufen, indoles
It is pungent（Indocin）, fluorine is than brufen, phenoxy group brufen, naproxen, Nabumetone（Nabumetone）, piroxicam（Scorching pain is liked
Health）, phenylbutazone, diclofenac sodium, Fen Luofen, Ketoprofen, ketorolac, four clofenamic acidses, sulindac, tolmetin etc. and/
Or glucocorticoid such as prednisone, hydrocortisone, dexamethasone etc. and/or immunosuppressor such as methotrexate (MTX), take fluorine rice
Spy, salicylazosulfapyridine, hydroxychloroquine sulfate etc. and/or biological agent such as Enbrel, benefit match general, Qiang Ke, class gram, Xiu Meile etc.
Deng.
Preferably application mode administration in addition of the invention, the present invention are applied to mouse by maximum dosage-feeding and are used ointment of the present invention one
Interior total dosage reaches 15.84g crude drugs/kg weight, still safe.Preferred dose scope is 0.01-15.84g crude drugs/kg bodies
Weight/day.
The numbness side of the disappearing crude drug content of the embodiment of the present invention 1：0.48g crude drugs/g medicinal extract；0.528 g crude drugs/ml medicinal extract.
Numbness people from the side of disappearing quantity：5g medicinal extract/day × 0.48g crude drugs/g medicinal extract=2.4g crude drugs/people/day；By 60/kg bodies
Re-computation：2.4g crude drugs/people/day ÷ 60kg/ people=0.04g crude drugs/kg body weight/days（People）
To mouse：Administration concentration：0.528g crude drugs/ml medicinal extract；
Volume is administered：0.1ml/10g weight=10ml/kg, administration number of times：3 times；
Day administration total amount：0.528g crude drugs/ml medicinal extract × 10 × 3=15.84g crude drugs/kg/ days.
Chinese medicine composition of the present invention can be prepared according to this field conventional method.Using Chinese medicine composition of the present invention, use
This field routine techniques and/or auxiliary material, those skilled in the art can prepare other drugs and preparation.
Chinese medicine composition of the present invention, pharmaceutical composition, drug and preparation can be used for treating rheumatoid arthritis, can disappear
Swollen, analgesic.Particularly, inventor has found Chinese medicine composition of the present invention, with tripterygium wilfordii, saltcake, Rhizoma Chuanxiong, frankincense（System）、
Myrrh（System）Combination compare, there is more significant detumescence, analgesic effect.
Embodiment
The following example only for illustrating invention, is not construed as the scope of limitation invention.
Embodiment 1
Take 60 grams of tripterygium wilfordii, 60 grams of saltcake, 30 grams of Rhizoma Chuanxiong, 30 grams of the root of Dahurain angelica, 15 grams of rhubarb, frankincense（System）15 grams, myrrh
（System）15 grams, 15 grams of peppermint, with water boiling and extraction 3 times, merging filtrate filters, and concentration adds in vegetable oil, adds up to 500 grams, system
It into ointment or is equably applied on non-woven fabrics and ointment medicine is made.
Embodiment 2
Take 120 grams of tripterygium wilfordii, 120 grams of saltcake, 60 grams of Rhizoma Chuanxiong, 60 grams of the root of Dahurain angelica, 30 grams of rhubarb, frankincense（System）30 grams, do not have
Medicine（System）30 grams, 30 grams of peppermint, tripterygium wilfordii, rheum officinale, the root of Dahurain angelica every time with 8 times amount 75% alcohol refluxs extract 3 times, every time 1 it is small when,
Merge extracting solution, filtration, filtrate recycling ethanol is spare, and the another device of the dregs of a decoction preserves.Rhizoma Chuanxiong, frankincense, three taste of myrrh add 6 times of water, extraction
When volatile oil 8 is small, volatile oil is collected, liquid filtration is spare；The dregs of a decoction of three taste such as Rhizoma Chuanxiong merge the dregs of a decoction of three tastes such as tripterygium wilfordii,
Saltcake is added in, adds 6 times of water, when decoction 1 is small, filtration, filtrate merges other above-mentioned liquids, is concentrated into relative density 1.055-
1.075（75℃）, 100 mesh sieves filter, spare.Peppermint is through steam distillation, then freezes, and the de- brain in part is processed to obtain 0.3ml thin
Lotus vegetable oil.After volatile oil, peppermint oil dementholized and the ethyl hydroxy benzoate of the three taste medicine such as more than Rhizoma Chuanxiong is taken to be dissolved with ethanol in proper amount, by ethyl alcohol
Liquid, appropriate sodium carboxymethylcellulose are added in glycerine, after fully stirring well, add in the liquid of above-mentioned concentration, and water is added to adjust total amount
To 1000g, continue to stir, gelling agent is made.
Embodiment 3
Take 70 grams of tripterygium wilfordii, 55 grams of saltcake, 25 grams of Rhizoma Chuanxiong, 30 grams of the root of Dahurain angelica, 15 grams of rhubarb, frankincense（System）15 grams, myrrh
（System）15 grams, 15 grams of peppermint, 10 grams of Radix Glycyrrhizae are extracted with 75% ethyl alcohol, are filtered, and concentration obtains extract.Vegetable oil is added in, is made
Ointment.
Embodiment 4
Take 50 grams of tripterygium wilfordii, 65 grams of saltcake, 35 grams of Rhizoma Chuanxiong, 30 grams of the root of Dahurain angelica, 15 grams of rhubarb, frankincense（System）15 grams, myrrh
（System）15 grams, 15 grams of peppermint, drying and crushing cross 120 mesh sieves.Coating agent is made in gained powder with conventional Chinese medicine method.
The Chinese medicine composition or drug of the present invention can treat rheumatoid arthritis.The particularly gelling agent of embodiment 2, than
The preparation of embodiment 1 has preferably detumescence, analgesic effect.
The 1 numbness side of disappearing of the present invention of test example acts on the easing pain and diminishing inflammation of mouse
1. test material
1.1 test medicine：The numbness side of disappearing（The ointment prepared according to 1 method of embodiment）:0.48g crude drugs/g ointment;Batch
Number：080312；By China Academy of Sciences Guanganmen Hospital Daxing, Drug Manufacturing Room provide.Voltarol：The limited public affairs of Beijing Novartis
Department's production, 2011.01 2008.02X1012 of lot number.Dimethylbenzene：52952 chemical plant of Beijing provides, lot number：870330；
Blank control medicine：Bare substrate lot number：080312, by China Academy of Sciences Guanganmen Hospital, Drug Manufacturing Room provide.
1.2 test dose:The numbness side of disappearing（0.48g crude drugs/g）, be grown up clinical daily 5g, i.e., daily clinical application amount is
2.40g crude drugs/d, in terms of the weight 60kg that is grown up, average dosage is 0.04g crude drugs/kg/d.This test mice is basic, normal, high
Three dosage groups, be set to 0.2,0.4,0.8g crude drugs/kg, above-mentioned dosage is 5,10,20 times of clinical medicine dose（Press body
Re-computation）, positive control drug Vitalin Emulgel group（0.016ml ointment/kg）10 times of people's clinical application amounts are equivalent to, it is negative right
According to medicine to the matrix of middle dose group equivalent.
1.3 animal：KM mouse are provided by Chinese military medicine academy of sciences animal center.Meet cleaning grade standard, the quality certification
Number：SCXK- (army) 2002-001.Normal raising supplies examination after 3 days.
2. test method and result
The influence of 2.1 pairs of thermostimulation induced mice pain（Hot plate method）
KM mouse 70, female, 18~22g of weight are raised 3 days after buying, and choose Basic Pain Threshold between 5~30s
Mouse 50（Since mouse is placed in hot plate temperature for 55 ± 0.5 DEG C up to mouse lick metapedes time based on the threshold of pain）
It is randomly divided into 5 groups, i.e. bare substrate control group, the basic, normal, high dosage group in the numbness side of disappearing, Voltarol positive controls.Daily backside
Medicine 1 time, blank control group apply the matrix of equivalent, continuous 6 days, 2 × 1.5cm of antedorsal shaving are administered.After the last administration, medicine is measured
Afterwards 0.5,1.0,1.5, the mouse threshold of pain of 2.0h, calculates the threshold of pain and improves percentage, more each group difference.It the results are shown in Table 1.
Influence of the 1 numbness side of disappearing of table to thermostimulation induced mice pain
Note：Compared with bare substrate group,*P<0.05,**P<0.01,***P<0.001
As seen from Table 1, the basic, normal, high dosage each group mouse in the numbness side of disappearing can improve 0.5~2h mouse threshold of pain to some extent
Improve percentage, the P compared with bare substrate group<0.05~0.001.Voltarol can also significantly improve the mouse threshold of pain and improve percentage
Rate, the P compared with bare substrate group<0.001.
The influence of 2.2 paraxylene induced mice auricle edemas
Take KM mouse 50,18~22g of weight, female is randomly divided into 5 groups, i.e., bare substrate control group, the numbness side of disappearing it is low,
Middle and high dosage group, Voltarol positive controls, then daily coating 1 time, blank control group apply the matrix of equivalent, continuous 6 days,
After last dose 1h, by 50 μ l drops of dimethylbenzene in mouse right ear two sides, left ear is not applied as control, by mouse cervical dislocation after 0.5h
It puts to death, lays round auricle at the same position of left and right ear respectively with diameter 6mm card punch, precision balance is weighed, with left and right auricle
The difference of weight is as swelling；（Cause scorching auris dextra weight-left ear weight of control）The left ear weight=ear swelling rate of/control（%）, compare
Each group difference.It the results are shown in Table 2.
The influence of the 2 numbness side of disappearing paraxylene induced mice auricle edema of table
Note：Compared with bare substrate group,*P<0.05,**P<0.01,***P<0.001
Table 2 the result shows that：The basic, normal, high dosage in the numbness side of disappearing can mitigate the swelling of auricle of caused by dimethylbenzene xylene inflammation mouse to some extent
Expansibility and swelling rate（P<0.05~P<0.01）, Voltarol also can significantly mitigate the ear swelling degree and swelling rate of mouse（P<0.01
~P<0.001）.
3. it discusses
The basic, normal, high dosage each group animal in the numbness side of disappearing can work after being administered 30 minutes, and with administration time extension and
The increase of dosage, the mouse threshold of pain improve percentage be also continuously improved, to 1.5 it is small when reach pharmaceutically-active peak, afterwards
Drug action is maintained or slightly weakened.Prompt the numbness side of disappearing that there is apparent analgesic activity to mouse.
The acute inflammation models such as the basic, normal, high dosage each group animal paraxylene induced mice auricle edema in the numbness side of disappearing also have
Obvious inhibiting effect, in 20 times of clinical application amounts in the range of, with the increase of dosage, swelling inhibitory action adds therewith
By force, the intensity of inhibitory action is：Heavy dose of group>Middle dose group>Small dose group.Prompt the numbness side of disappearing that there is significant anti-inflammation detumescence
Effect.
4. conclusion
This experimental result prompting numbness side of disappearing can improve the scorching mouse swelling degree of cause, improve tolerance of the mouse to thermostimulation
Degree has apparent anti-inflammatory, swollen, analgesic effect of dispelling.
The 2 numbness side of disappearing of the present invention of test example induces rats with arthritis therapeutic effect to CII
1. test material
1.1 test medicines and reagent：
The numbness side of disappearing（The ointment prepared according to 1 method of embodiment）:0.48g crude drugs/g ointment, lot number：080312, by
China Academy of Sciences Guanganmen Hospital Daxing Drug Manufacturing Room provide；Voltarol：Novartis Pharma AG produces, lot number
2011.012008.02X1012；Blank control medicine：Bare substrate lot number：080312, by China Academy of Sciences Guanganmen Hospital
Drug Manufacturing Room provide；II Collagen Type VIs（CII）:(sigma is public by Bovine Immunization Grade TypeII Collage
Department)；Incomplete Freund's adjuvant（IFA）:Incomplete Freund ' Adjuvant (sigma companies).
1.2 experiment dosages:The numbness side of disappearing（0.48g crude drugs/g）, be grown up clinical daily 5g, i.e., daily clinical application amount is
2.40g crude drugs/d, in terms of the weight 60kg that is grown up, average dosage is 0.04g crude drugs/kg/d.This test mice is basic, normal, high
Three dosage groups, be set to 0.25,0.50,1.00g crude drugs/kg, above-mentioned dosage for clinical medicine dose 6.25,12.5,
25 times（According to the weight）, positive control drug Vitalin Emulgel group（0.016ml ointment/kg）It is equivalent to the clinical use of 12.5 times of people
Dose, negative control medicine is to the matrix of middle dose group equivalent.
1.3 animal：SD rats 80,160-180g/, half male and half female, sub-cage rearing.Rat is purchased from Chinese drug biology
Product calibrating institute's Laboratory Animal Resource center is bought, quality certification number：SCXK (capital) 2005-0004.Rat feeding is in China
Gate of Pervasive Peace Hospital Clean grade animal housing of medical courses in general institute（Quality certification number：SYXK (capital) 2005-0001）, normal raising 3 days.
1.4 instrument：PV-200 toes capacity measurers（Chengdu TME Technology Co., Ltd. produces）.
2. test method
2.1CII albumen makes rat arthritis model
IFA is placed in test tube, under ice bath, every test tube stirs evenly IFA's with 10000 revs/min of rotating speed with vortex mixer
Meanwhile the CII of equivalent is slowly added dropwise into pipe.Stir evenly 3 minutes every time, every test tube stirs evenly 3-4 times, until mixing extremely
（Emulsus）Oil-in-water shape is instilled a drop suspension in water without until scattering.Under ice bath, by the IFA+CII liquid of mixing
To 70 rat root of the tail portions with after 75% alcohol disinfecting, injection 0.2ml/ only, is carried out immune for the first time.It is used after 7 days immune for the first time
Same method carries out rat half to measure immune, i.e. every rat root of the tail portion injection IFA+CII liquid 0.1ml/ for the second time.8 days
It will delineate and mark at 0.5-1.0cm on all rat double lower limb ankle-joints afterwards.Start to measure all rats with foot volumetric type meter after 9 days
Sufficient volume and articular index.The 11st day rat 50 for choosing double lower limb swelling, is randomly divided into 5 groups, every group of 10 rats.Start
It is grouped double lower limb coating：Matrix group（Matrix）；Low dose group is administered（The numbness side of disappearing）；Middle dose group（The numbness side of disappearing）；High dose group（Numbness
The side of disappearing）；Positive controls（Voltarol）；Other 10 are not normal group by immune rat.Rat coating 1 time is given daily, even
Continuous coating 18 days.Rat foot volume was measured per 3-4 days once, observation joint pathology integration and pathology damage state.
2.2 rat articulars damage pathology integration evaluation criteria：
◆ 0 point：Joint structure is normal, and joint space is uniform, and cartilage, bone are not damaged, synovial tissue's no inflammation, hyperplasia etc.；
◆ 1 point：Joint structure is normal, there is synovial hyperplasia, and blood vessel number increases, it is seen that a small amount of cell infiltration, it is soft
The light damage of bone surface layer；
◆ 2 points：Articular cartilage has apparent erosion damage, and pannus is formed, and cell infiltration is apparent, no osteoclasia,
Joint space is normal；
◆ 3 points：There is substantial amounts of pannus to be formed, it is seen that the erosion damage of extensive cartilage, it is seen that osteoclasia, joint
Structure goes to pot.
2.3 pathology
The ankle-joint sample of each group rat is fixed using formalin, 5%HNO3After decalcification 1 week, alcohol is dehydrated step by step, and two
Toluene is transparent, paraffin Bao Li, section, HE dyeing.Light Microscopic observation pathological examination.
3. result
The 3.1 numbness sides of disappearing induce CII the improvement result of rats with arthritis foot volume
It it can be seen that, is immunized from table 1 and 2 in first time（CII induces arthroncus）During rat 9 days, rat articular is opened
There is swelling in beginning, normal group and immune group each group（Matrix group, low dosage, middle dosage, high dose, the positive being immunized before administration
Group rat）Compare, start difference occur；When being immunized 11 days, there is obvious tumefaction in rat articular, and normal group goes out with immune each group
Existing notable difference（P<0.001）.
1 CII of table induces arthritis female rats foot volume determination
Each group is compared with matrix group：*P<0.05 **P<0.01 # P<0.001
2 CII of table induces arthritis male rat foot volume determination
Each group is compared with matrix group：*P<0.05 # P<0.001
In rat immunity 11 days, we selected the rat of arthroncus to start grouping administration, are administered once daily, continuously
Administration 17 days measures weekly 2 sufficient volumes, observes and records arthroncus degree.
The 3 numbness side of disappearing of table induces arthritis female rats to CII（Left side）Sufficient volume improvement result
Each group is compared with matrix group：*P<0.05 **P<0.01 #P<0.001
The 4 numbness side of disappearing of table induces arthritis female rats to CII（Right side）Sufficient volume improvement result
Each group is compared with matrix group：*P<0.05 **P<0.01 #P<0.001
The 5 numbness side of disappearing of table induces arthritis male rat to CII（Left side）Sufficient volume improvement result
Each group is compared with matrix group：*P<0.05 **P<0.01 #P<0.001
The 6 numbness side of disappearing of table induces arthritis male rat to CII（Right side）Sufficient volume improvement result
Each group is compared with matrix group：*P<0.05 **P<0.01 #P<0.001
It will be seen that each administration group rat articular swelling is mitigated from table 3,4,5,6, in administration 10 days（Exempt from
Epidemic disease 21 days）When, the numbness side of disappearing high dose group and positive control（Voltarol）Group rat articular swelling is substantially reduced, and either female is gone back
It is that the sufficient volume of male rat is below the matrix group with the time（P<0.05）, it is big to illustrate that the numbness side of disappearing induces arthritis for CII
The arthroncus of mouse has certain mitigation effect.
The 3.2 numbness sides of disappearing induce rats with arthritis arthroncus pathological change to CII
3.2.1 rat ankle joint degree of injury pathology integral result
7 are shown in Table according to " rat articular damage pathology integration evaluation criteria " detection rat articular damage pathology integral result.
The improvement result that the 7 numbness side of disappearing of table integrates CII rats with arthritis joint injuries pathology
| Grouping/joint integration | n | X±SD |
| Matrix group | 10 | 3.00±0.00 |
| Low dose group | 10 | 2.30±1.16 |
| Middle dose group | 10 | 2.50±1.08 |
| High dose group | 10 | 1.90±1.45 |
| Positive controls | 10 | 1.90±1.29 |
| Normal group | 10 | 0.00±0.00# |
Each group is compared with matrix group：#P>0.001
As can be seen from Table 7, CII induces rat arthritis（Model）The arthroncus of matrix group is apparently higher than normal group（P
>0.001）；Administration each group acts on the arthroncus of the CII rats with arthritis induced, but is not statistically significant.
3.2.2 joint pathology observes result
Om observation result is as follows：
Normal group：Joint structure is normal, and synovial tissue is without congestion and edema, and synovial cell is without hyperplasia phenomenon, no blood vessel
Screen is formed, and articular cartilage surface is smooth, no peeling-off phenomenon, and bone trabecula size, arrangement are normal under articular cartilage.
Matrix group：In addition to female No. 2 only slight synovial cell proliferation, the visible a large amount of synovial membranes of each ankle-joint of remaining rat are thin
Born of the same parents' hyperplasia, synovial cell is disorganized, synovial tissue's congestion and edema, blood capillary proliferation, and visible a large amount of cell infiltrations；Increase
Raw synovial tissue forms villiform, stretches to articular cavity depths or creeps to cartilage surface, forms pannus.It is covered in pannus
Under being denatured of the visible apparent cartilage textura epidermoidea in cartilage surface layer, necrosis, and visible pannus stretches into bone tissue, causes
Apparent osteoclasia, joint structure are substantially destroyed, and have articular cartilage and the synovial tissue of de- stripping in articular cavity.
Positive controls：5 joint pathologies change with matrix group；2 joints have no apparent pathological change；2 joints can
Seeing has slight synovial cell proliferation；1 is seen there is severe synovial cell proliferation, and the synovial tissue of hyperplasia forms villiform, stretches to pass
Chamber depths is saved, and is creeped to cartilage surface, forms pannus.
High dose group：The pathological change for having 6 joints is light compared with matrix group, and 3 joints have no apparent pathological change；1 pass
Save it is visible have slight synovial cell proliferation, and with a small amount of cell infiltration.
Middle dose group：There is 8 joints are visible to have typical arthritis pathological change, pathological change is the same as matrix group；1 joint
Have no apparent pathological change；1 joint is visible slight synovial cell proliferation, with a small amount of cell infiltration.
Low dose group：There is 7 joints are visible to have typical pathological change, pathological change has no bright with matrix group, 1 joint
Aobvious pathological change, 2 joints are visible slight synovial cell proliferation, with a small amount of cell infiltration, the light damage of cartilage surface layer.
Pathology impression
Matrix group rat is visible apparent arthritis pathological change, and the pathological change of high dose group is bright compared with matrix group
It is aobvious to mitigate.In, the pathological change of low dose group and model group no significant difference.
4. brief summary
The arthroncus that the numbness side of disappearing high dose group rat induces for CII can be seen that by above-mentioned experimental result（Foot holds
Product）, joint injury degree pathology integration have certain mitigation effect；Pathological observation also turns out this point, i.e. high dose group rat
Arthritis cellular infiltration is reduced, and blood capillary proliferation, synovial tissue's congestion and edema, synovial cell proliferation make moderate progress.
The inflammatory symptoms for the rats with arthritis that the prompting Chinese medicine compound prescription numbness side of disappearing is induced mitigating CII collagens have certain effect.
3 local application's toxicity test of test example
1. test material
1.1 test medicine
The numbness side of disappearing:0.48g crude drugs/g medicinal extract, by China Academy of Sciences Guanganmen Hospital, centers for making of pharmaceutical preparations provide, lot number
080312.Bare substrate medicine：By China Academy of Sciences Guanganmen Hospital, Drug Manufacturing Room provide, lot number 080312.Voltarol：Beijing
Nova Lab SDN. BHD produces, lot number X1012.
1.2 reagent
2.4- dinitro-chloro-benzene：Beijing Chemical Plant, three-level, lot number 710710.
Acetone：Beijing chemical reagents corporation analyzes pure, lot number 20050104.Be made into during experiment with acetone 1% sensitization it is dense
Degree and 0.1% excitation concentration.
1.3 animal：
New zealand rabbit：Center, (quality certification number are cultivated purchased from Beijing rich and powerful people experimental animal：SCXK (capital) 2005-0009), just
Often raising, after a week for examination.
Cleaning grade Hartley cavys：Purchased from Nat'l Pharmaceutical ＆ Biological Products Control Institute's Laboratory Animal Resource center, the quality certification
Number：SCXK (capital) 2005-0004, the cavy feed quality certification：SCXK- (army) 2007-005.Ad lib is drunk water, and normally raises 5
For examination after it.
1.3 rearing conditions：
New zealand rabbit is normally raised in China Academy of Sciences Guanganmen Hospital cleaning grade animal housing, quality certification number：SYXK
(capital) 2005-0001.
Cleaning grade Hartley cavys are normally raised in cleaning grade animal housing of the Capital University of Medical Sciences, quality certification number：SYXK (capital)
2004-0007。
2 experimental methods and result
2.1 Skin Irritation Test
Take healthy new zealand rabbit 8,2.0~2.5kg of weight, half male and half female.Every rabbit will when before administration 24 is small
Animal backbone both sides 50cm2Skin unhairing simultaneously cleans, this experiment using androgynous left and right sides skin self-contrast, be divided into administration area with
Check plot, left side go to hair-fields to apply the numbness side of disappearing 1ml, and right side check plot applies equivalent matrix and makees blank control, then with gauze and adhesive plaster
Fixed, every animal sub-cage rearing divides single-dose（4,3 times/day, when small per minor tick 4）And multiple dosing（4,1 time/
My god, continuous 7 days）Irritant test.
2.1.1 single-dose (acute) irritation test
Be administered 24 it is small when after with warm water remove liquid, in remove tested material 1,24,48,72 it is small when after visually observe animal
Local reaction is observed continuously 7.It presses《Study of tcm new drug guide》In related skin irritation reaction standards of grading (see attached list
1st, 2) record stimulates score value, while the skin of clip medicine-feeding part does histopathologic examination, the results are shown in Table 1.
2.1.2 multiple dosing irritation test
Successive administration 7 days, when last dose 24 is small after, with warm water remove drug, observation removal drug after 1 to 7 day apply
The response situation of medicine area skin scores with reference to single-dose irritation test method, while the skin of clip medicine-feeding part
Fu Zuo histopathologic examinations, the results are shown in Table 1.
1. skin wound repair standards of grading of subordinate list
2 skin irritatin intensity evaluation standard of subordinate list
Table 1：The numbness side of disappearing is to the average response value of skin irritatin
React average value：（Erythema forms total score+oedema and forms total score）/ total number of animals
The result shows that standards of grading and irritation intensity evaluation are reacted according to skin irritation, it is single or multiple to be coated with numbness
The side of disappearing and matrix do not occur phenomena such as erythema, oedema to rabbit normal skin, also have no cutaneous pigmentation or pachylosis
Situations such as occur.Skin irritatin average response value is 0.Stimulus intensity is evaluated as 0.Histopathological examination also shows：Check plot
It is complete with visible epidermis under administration area's mirror, it has no necrosis, come off, have no hyperemia and inflammatory cell infiltration, have no epithelium foot cell
Thickening and other lesions.This experiment prompting the numbness side of disappearing it is nonirritant to normal intact skin (by mean scores 0-0.49 is evaluated as
It is nonirritant), see pathological tissue photo.
2.2 hypersensitive test
Cleaning grade Hartley cavys 30, half male and half female are taken, 250~300g of weight is randomly divided into 3 groups:Bare substrate
Group, the numbness side of disappearing group, positive controls (2,4- dinitro-chloro-benzenes), scrape off guinea pig back diamond wool when before experiment 24 is small,
Hair-fields about 3 × 3cm is removed per side2, the following two steps progress of experiment point.
2.2.1 sensitization contact is tested:Every animal sub-cage rearing takes the numbness side of disappearing 0.2g to be coated on the right side of cavy and goes to hair-fields, hold
It is continuous 6 it is small when, in the 7th and 14 day, be in kind respectively repeated once, altogether three times, bare substrate group and positive controls drug,
Dosage method is same as above.
2.2.2 contact test is excited:14 days after last dose sensitization, the test medicine 0.2g of each group is applied to the cavy back of the body
Hair removal section on the left of portion, 6 it is small when after with distilled water flushing back residual drug, observe at once, then when 24,48,72 is small again
Cutaneous anaphylaxis and general reaction situation are observed, is pressed《Study of tcm new drug guide》In the scoring mark in relation to cutaneous anaphylaxis
Accurate (seeing attached list 3,4) is evaluated, and the results are shown in Table 2.
The standards of grading of 3. cutaneous anaphylaxis degree of subordinate list
React average value：（Erythema forms total score+oedema and forms total score）/ total number of animals
4 hypersensitive evaluation criterion of subordinate list
Sensitization incidence：It will appear from the animal number of cases of erythema, oedema or systemic anaphylaxis（No matter mild degree
Weight）, except in animal subject sum
The 2. numbness side of disappearing of table is on the anaphylactoid influence of cavy
React average value：（Erythema forms total score+oedema and forms total score）/ total number of animals
The experimental results showed that:The tested area's excitation 6 of positive drug 1-CHLORO-2,4-DINITROBENZENE group guinea pig skin occurs slight when small
Erythema, no oedema, when 24,48,72 is small after there is erythema, oedema, sensitization rate 100%, blank control group and the group excitation of the numbness side of disappearing
Sensitization phenomenon and general reaction are had no when 6,24,48,72 is small afterwards, by 4 hypersensitive standards of grading of subordinate list, can determine whether the numbness side of disappearing
Without sensitization, allergy effect is not generated.
3. conclusion
Cavy intact skin single and the numbness side of disappearing smearing repeatedly is given, do not find skin histology exception, prompt the numbness side of disappearing
To no skin irritation.Cavy allergic experiment result also shows Bi Xiaofang local applications and does not cause allergic reaction simultaneously, prompts numbness
The side's of disappearing clinical application safety.
Claims (12)
1. Chinese medicine composition, it is characterised in that
Its by 50 to 70 grams of tripterygium wilfordii, 55 to 65 grams of saltcake, 20 to 40 grams of Rhizoma Chuanxiong, 20 to 40 grams of the root of Dahurain angelica, 10 to 20 grams of rhubarb,
10 to 20 grams of frankincense, 10 to 20 grams of myrrh, 10 grams of peppermint to 20 restrain into.
2. according to the Chinese medicine composition of claim 1, wherein
It is by 60 grams of tripterygium wilfordii, 60 grams of saltcake, 30 grams of Rhizoma Chuanxiong, 30 grams of the root of Dahurain angelica, 15 grams of rhubarb, 15 grams of frankincense, myrrh 15
Gram, peppermint 15 restrains into.
3. the preparation method of the Chinese medicine composition of claim 1 or 2, it is characterised in that：
(1) each Chinese medicine material is directly mixed, obtains Traditional Chinese drug mixture；Optionally and
(2) mixture obtained by water and/or alcohol extracting above-mentioned (1), obtains Chinese medical extract.
4. it is mixed according to the Traditional Chinese drug mixture prepared by the method for claim 3 or Chinese medical extract or containing their Chinese medicine
Object or Chinese medical extract.
5. the purposes of the Traditional Chinese drug mixture or Chinese medical extract in claim 4 is used to prepare the outer for the treatment of rheumatoid arthritis
Use drug.
6. the drug of rheumatoid arthritis is treated, using the Traditional Chinese drug mixture of claim 4 or Chinese medical extract as active ingredient.
7. treat the combination drug of rheumatoid arthritis, Traditional Chinese drug mixture or Chinese medical extract conduct including claim 4
Effective component of chinese medicine and NSAIDs and/or DMARDs class Western medicine active ingredients.
8. treating the pharmaceutical composition of rheumatoid arthritis, the drug including claim 6 or 7 is as active ingredient and pharmaceutically acceptable
Carrier, diluent or excipients.
9. treat the pharmaceutical preparation of rheumatoid arthritis, the pharmaceutical composition including claim 8.
10. the pharmaceutical preparation of claim 9 is solution, gelling agent, lotion, liniment, coating agent, spray, creme, cream
Agent or patch.
11. the pharmaceutical preparation of claim 10 is gelling agent.
12. the preparation method of the pharmaceutical preparation of claim 10 or 11, Traditional Chinese drug mixture or Chinese medicine including claim 4
Extract is mixed with pharmaceutically acceptable matrix, is optionally combined with Western medicine described in claim 7；Chinese medical extract therein is by following
Method is prepared：
I) tripterygium wilfordii, rheum officinale, the root of Dahurain angelica are extracted with ethyl alcohol, and filtration, filtrate recycling ethanol is spare, and the another device of the dregs of a decoction preserves；
Ii) Rhizoma Chuanxiong, frankincense, three taste extracting in water volatile oil of myrrh, collect volatile oil, and liquid filtration is spare；
Iii) Rhizoma Chuanxiong, frankincense, the dregs of a decoction of three taste of myrrh merge tripterygium wilfordii, rheum officinale, the dregs of a decoction of three taste of the root of Dahurain angelica, add in saltcake, use water
It decocts, filtration, filtrate merges other above-mentioned liquids, concentrates, and filtration is spare；
Iv) take more than Rhizoma Chuanxiong, frankincense, three taste medicine of myrrh volatile oil, from peppermint extract peppermint oil dementholized dissolved with ethyl alcohol, by second
Alcohol liquid is mixed with the liquid of above-mentioned concentration.
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