CN104352633B - A kind of preparation method of pharmaceutical composition that treating osteoarthropathy - Google Patents
A kind of preparation method of pharmaceutical composition that treating osteoarthropathy Download PDFInfo
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Abstract
The invention belongs to field of traditional Chinese medicine preparations, and in particular to a kind of for treating the preparation method of the pharmaceutical composition of osteoarthropathy.Preparation method of the present invention: it is used as medicine with rhizoma cibotii, Herba Epimedii, Radix Angelicae Pubescentis, the rhizome of davallia, teasel root, herba taxilli, Caulis Spatholobi, Rehmannia glutinosa, radix aucklandiae and Olibanum volatile oil, Myrrha volatile oil and psoralea corylifolia processed product and is prepared.The pharmaceutical composition that the method is prepared not only effectively treats Osteoarthritis and lumbar muscle strain caused by kidney deficiency and liver, qi depression to blood stasis, the resistance of train of thought numbness, and effectively reduces the hepatotoxicity wind agitation side effect of original product, has better clinical therapeutic efficacy.
Description
Technical field
The invention belongs to field of traditional Chinese medicine preparations, and in particular to a kind of for treating the preparation of the pharmaceutical composition of osteoarthropathy
Method.
Background technique
Osteoarthritis (osteoarthritis, OA) is a kind of common chronic joint diseases, and major lesions are to close
Save the retrogression and secondary osteoproliferation of cartilage.Degenerative osteoarthropathy refers to a series of degenerative diseases that human body occurs
Become, such as arthralgia, stiff and limitation of activity, is a kind of comprehensive disease.It is in 50 years old or more crowd, and disease incidence is only
Inferior to heart disease, the disease incidence in over-65s crowd is up to 60-90%, especially brings pole to the life of the middle-aged and the old
Big pain and inconvenience.
Lumbar muscle strain is also one of the most common type pain in Low Back Pain disease, is often used as the chronic low back of no organic change
The general name of pain.Lumbar muscle strain is as caused by a lot of reasons, and the prolonged and repeated breaking-out of pain, waist is ached or distending pain.It is clinical
It shows as, hyperactivity hyperkinesia, then pain exacerbation of bending over too long, straight waist is difficult, and sombre, wet weather can aggravate illness.
For above-mentioned Osteoarthritis and lumbar muscle strain symptom, Chinese patent CN1931277A disclose it is a kind of treat bone pass
Save the pharmaceutical composition of disease.Described pharmaceutical composition by rhizoma cibotii, Herba Epimedii, Radix Angelicae Pubescentis, the rhizome of davallia, teasel root, psoralea corylifolia, herba taxilli,
Caulis Spatholobi, Rehmannia glutinosa, radix aucklandiae, olibanum, 12 taste Chinese medicinal composition of myrrh have and tonify the liver and kidney, nourshing blood and promoting blood circulation, relaxing tendons and activating collaterals, reason
The functions such as gas analgesic, are mainly used for kidney deficiency and liver, stagnation of QI and blood, the diseases such as Osteoarthritis, lumbar muscle strain caused by obstruction of meridians and collaterals.
And animal and clinical test are shown, which has exact curative effect, and obtains many patients to Osteoarthritis, lumbar muscle strain
Favor.
However, said medicine finds that there are certain hepatic injuries during clinical use, for this purpose, before guaranteeing drug effect
It puts, it is problem in the urgent need to address that being carried out by process modification to it, which reduces hepatic injury research,.
Summary of the invention
For this purpose, technical problem to be solved by the present invention lies in the drugs for treating osteoarthropathy in the prior art to exist centainly
Hepatic injury adverse reaction the problem of, and then develop a kind of hepatotoxicity wind agitation and damage drug that is lower, can effectively treating osteoarthropathy
The preparation method of composition.
In order to solve the above technical problems, the preparation method of the pharmaceutical composition for the treatment of osteoarthropathy of the present invention, packet
Include following steps:
(1) olibanum 154-520 parts by weight, the mixing of myrrh 154-520 parts by weight are taken, water is added to carry out steam distillation extraction,
Volatile oil is collected, customary adjuvant is added, volatile oil clathrate compound is made, it is spare；
(2) psoralea corylifolia 154-520 parts by weight are taken, extracting in water collects the dregs of a decoction after discarding Aqueous extracts, obtains psoralea corylifolia processing
Product, it is spare；
(3) rhizoma cibotii 290-864 parts by weight, Herba Epimedii 154-520 parts by weight, Radix Angelicae Pubescentis 154-520 parts by weight, the rhizome of davallia are taken
224-692 parts by weight, teasel root 290-864 parts by weight, herba taxilli 290-864 parts by weight, Caulis Spatholobi 154-520 parts by weight, radix rehmanniae preparata
Yellow 692-2076 parts by weight, radix aucklandiae 154-520 parts by weight, mix with the spare psoralea corylifolia processed product, crush, described in addition
Clinically-acceptable oral preparation is made according to common process in spare volatile oil clathrate compound and customary adjuvant.
Further, the preparation method of the pharmaceutical composition, includes the following steps:
(1) 346.1 parts by weight of olibanum, the mixing of 346.1 parts by weight of myrrh are taken, adds water to carry out steam distillation extraction, collects
Volatile oil is added customary adjuvant and volatile oil clathrate compound is made, spare；
(2) 346.1 parts by weight of psoralea corylifolia are taken, extracting in water collects the dregs of a decoction after discarding Aqueous extracts, obtains psoralea corylifolia processing
Product, it is spare；
(3) 576.8 parts by weight of rhizoma cibotii, 346.1 parts by weight of Herba Epimedii, 346.1 parts by weight of Radix Angelicae Pubescentis, 462.0 weight of the rhizome of davallia are taken
Measure part, 576.8 parts by weight of teasel root, 576.8 parts by weight of herba taxilli, 346.1 parts by weight of Caulis Spatholobi, 1384.2 parts by weight of Rehmannia glutinosa,
346.1 parts by weight of radix aucklandiae are mixed with the spare psoralea corylifolia processed product, are crushed, and add the spare volatile oil clathrate compound
And customary adjuvant, clinically-acceptable oral preparation is made according to common process.
Preferably, the preparation method of the pharmaceutical composition, includes the following steps:
(1) it takes the olibanum and myrrh to mix according to selected parts by weight, adds 8-12 times to measure water progress steam distillation and mention
It takes 8-15 hours, collects volatile oil, customary adjuvant is added, volatile oil clathrate compound is made, it is spare；
(2) psoralea corylifolia is taken according to selected parts by weight, adds 6-10 times to measure water and extracts 1-3 times, it is 0.5-2 hours each,
The dregs of a decoction are collected after discarding Aqueous extracts, are drying to obtain psoralea corylifolia processed product, it is spare；
(3) rhizoma cibotii, Herba Epimedii, Radix Angelicae Pubescentis, the rhizome of davallia, teasel root, herba taxilli, Caulis Spatholobi, radix rehmanniae preparata are taken according to selected parts by weight
Yellow, radix aucklandiae, mixes with the spare psoralea corylifolia processed product, crushes, and adds the spare volatile oil clathrate compound and routine is auxiliary
Material, is made clinically-acceptable oral preparation according to common process.
More preferably, the preparation method of the pharmaceutical composition, includes the following steps:
(1) it takes the olibanum and myrrh to mix according to selected parts by weight, 10 times of amount water is added to carry out steam distillation extraction
12 hours, volatile oil is collected, customary adjuvant is added, volatile oil clathrate compound is made, it is spare；
(2) psoralea corylifolia is taken according to selected parts by weight, adds 8 times of amount water to extract 2 times, 1 hour every time, discards Aqueous extracts
After collect the dregs of a decoction, be drying to obtain psoralea corylifolia processed product, it is spare；
(3) rhizoma cibotii, Herba Epimedii, Radix Angelicae Pubescentis, the rhizome of davallia, teasel root, herba taxilli, Caulis Spatholobi, radix rehmanniae preparata are taken according to selected parts by weight
Yellow, radix aucklandiae, mixes with the spare psoralea corylifolia processed product, crushes, and adds the spare volatile oil clathrate compound and routine is auxiliary
Material, is made clinically-acceptable oral preparation according to common process.
In the step (1), the customary adjuvant for preparing the volatile oil clathrate compound includes beta-cyclodextrin, water and ethyl alcohol.
In the step (1), the volatile oil, beta-cyclodextrin, water and ethyl alcohol mass ratio be 1:8-12:16-24:1-2.
The volatile oil, beta-cyclodextrin, water and ethyl alcohol mass ratio be 1:10:20:1.
The olibanum and/or myrrh are vinegar toast.
The invention also discloses the pharmaceutical compositions being prepared by the preparation method, and by aforementioned pharmaceutical compositions
Customary adjuvant is added, according to clinically-acceptable oral preparation made of common process.
Preparation method of the present invention improves on the basis of existing product, the medicine group of the treatment osteoarthropathy
It closes object to be made of medicinal materials such as psoralea corylifolia, the rhizome of davallia, teasel root, Herba Epimedii, Radix Angelicae Pubescentis, radix aucklandiae, Caulis Spatholobi and rhizoma cibotii, is mainly used for liver kidney
The diseases such as Osteoarthritis and lumbar muscle strain caused by deficiency, qi depression to blood stasis, the resistance of train of thought numbness.Preparation pharmaceutical composition of the present invention
Method be used as medicine with the volatile oil and psoralea corylifolia processed product of olibanum and myrrh, on the basis of keeping existing product drug effect, into
One step greatly reduces the hepatotoxicity wind agitation side effect of original product, so that product has better medical value and market prospects.
Experimental example
1 pharmacodynamic experiment of experimental example
1 material
1.1 animal
Hartley cavy, regular grade, SD rat, KM mouse are SPF grades, limited purchased from Beijing China Fukang biotechnology share
Company, quality certification number: SCXK (capital) 2009-0004；It raises in Tianjin University Of Traditional Chinese Medicine's Experimental Animal Center, keeps animal house peace
It is quiet, it divulges information, dry, 18~25 DEG C of room temperature, humidity 40%~70% freely ingests, drinks water.
1.2 test drug
New process pill prepared by embodiment 1；
Zhuanggu guanjie pill, China Resources Sanjiu Medical & Pharmaceutical Co., Ltd..
Experimental administration dosage refers to clinical people's dosage of zhuanggu guanjie pill: 12g/d, and each dosage in experiment is all pressed
Corresponding dosage is converted into according to different genera animal.
1.3 reagent
Aminoglucose hydrochloride capsule (Zhejiang Cheng Yi Pharmaceutical Co., Ltd)；
Papain, F127 (Sigma company)；
0.5% eosin stains liquid, haematoxylin dyeing liquid (military rattan Chemical Co., Ltd.)；
Fibroblast collagenase (MMP-1), Fibroblast collagenase inhibitor (TIMP-1) Elisa kit
(Cusabio)；TIMP-1 and MMP-3ELISA kit (Wuhan You Ersheng company)；UNIQ-10column Trizol total serum IgE
Extracts kit (Shanghai Sheng Gong bioengineering limited liability company)；mPromⅡReverse Transcription System
Reverse Transcriptase kit (Promega)；FastStart Universal CYBR Green kit (Roche)；BCA Protein Detection
Kit (Beyotime)；
Alkaline phosphatase (AKP), Bioengineering Research Institute is built up in Nanjing；
Benzalkonium bromide solution, Jiangxi Jingdone district pharmaceutcal corporation, Ltd；
Benzylpenicillin sodium for injection, middle promise medicine company (Shijiazhuang) Co., Ltd；
Glacial acetic acid, Tianjin Chemical Reagents Factory No.1；
Formaldehyde, Tianjin Fengchuan Chemical Reagent Science & Technology Co., Ltd.；
Dimethylbenzene, Tianjin Jinbei Fine Chemicals Co., Ltd..
1.4 instrument
JJ3000 electronic balance, JJ2000 precision electronic balance (Changshu Shuan Jie test equipment factory)；
YLS-Q4 ears swell card punch (Jinan Yi Yan development in science and technology Co., Ltd)；
HSS-1 (B) thermostatic bath (Chengdu Instruement Factory)；
MEK-6318K type cellanalyzer (Nihon Kohden Corporation)；
Nucleic acid-protein analyzer-DU800, Allegra 64R High-Speed Centrifuge (Beckman
Coulter)；Real-time RT-PCR System, Prism7500 (Applied Biosystem)；PCR-C1000 type PCR
Instrument, C1000Touch Thermal Cycler (Bio-Rad)；FlexStation3 multi-function microplate reader (Molecular
Devices)；1012 type of MODEL hybridizes case (SHEL LAB)；ASP300S Full automatic sealing dewaterer, RM2135 slicer
(Leica)；BMJ-1 biological tissue embedding machine (Tianjin Aviation Mechano-Electrical Co.)；Photomicroscope (OLYMPUS)；Ultrasonic Cell Disruptor
(UIBRA)；MEK-7222K Automatic Blood Cell Analyzer (Japanese photoelectricity industry Zhu Shi people's commune)；
IKA/T18 refiner (1KA)；KJ-201A type oscillator (Jiangsu Kangjian Medical Apparators Co., Ltd.)；
QL-901 type turbine mixer (its woods Bell's instrument manufacturing Co., Ltd, Haimen City)；
MilliQ pure water system (Millipore company).
2 methods
2.1 osteoarthritis treatment Effect studies
2.1.1 to the therapeutic effect of hareley cavy spontaneity osteoarthritis
Hartley cavy is taken, female is randomly divided into model group (control group), zhuanggu guanjie pill group (0.96g/kg) and sheet
5 inventive composition high dose (1.92g/kg), middle dosage (0.96g/kg), low dosage (0.48g/kg) dosage groups, every group 10
Only.After the spontaneous Osteoarthritis model of 5 monthly age cavys is formed, start to be administered, model group gives respective volume distilled water, continuously
Administration 4 weeks.Observation animal general state daily claims the weight of animals and appetite, after the last administration abdominal aortic blood on every Mondays, inspection
Survey the content of MMP-1, TIMP-1 in serum.Animal cuts shin bone and carries out histopathology to the articular cartilage between femur after putting to death
It learns and checks.
2.1.2 to the treatment of the rat bone arthritis chronica phase of Papain enzyme induction
Healthy male SD rat is selected, it is husky with 35% pool Lip river by the chloral hydrate anesthesia of 0.3g/kg weight intraperitoneal injection 10%
Nurse carries medicine body 4% papain of load and temperature sensitive type in-situ gel controlled release agent (pH5.7) is made, according to 0.05mL (1.6U)/pass
Dosage is saved, bilateral knee joint injection is placed in special incubator by modeling animal knee joint and with lower limb body, under 50 DEG C of environment
Insulation effect 2 hours, the damage of cartilage can be caused.It randomly selects 3/10 rat and carries out histopathologic examination, check mould
The reliability and success rate of type.
Selection modeling successful osteoarthritis rat 40, is randomly divided into model group, zhuanggu guanjie pill group, the present invention
Composition 2.20g/kg, 1.10g/kg and 0.55g/kg dosage group, every group 8.Optionally health male SD rat 8, as right
According to group.Zhuanggu guanjie pill and the present composition are according to setting dosage gastric infusion, continuous treatment 4 weeks, control group and model group
The isometric distilled water of stomach-filling.
After treatment end, anaesthetized by the chloraldurate of 0.3mL/100g weight intraperitoneal injection 10%, abdominal aorta takes
Blood.Left knee joint cavity is opened, with 1mL physiological saline repeated flushing 3 times, collects joint fluid, 3000r/min is centrifuged 10 minutes, takes
Supernatant, -20 DEG C save backup.With the surface cartilage of pocket knife scraping rat femur articular surface and tibial prosthesis face, weighing is put into
In 400 μ l Trizol, -80 DEG C of storages are spare, using real-time quantitative RT-PCR method measurement cartilage of rats iNOS,
Aggrecan and II mRNA of Collegan expression.Whole 10% formalin of right knee joint is taken to fix.Joint is measured with ELISA method
The content of liquid MMP-3 and TIMP-1, design of primers are shown in Table 1.
Conventional Trizol method extracts total serum IgE, and the purity of nucleic acid is estimated according to the value of OD260/OD280.Using Real-time
RT-PCR method measures II mRNA of cartilage of rats Aggrecan and Collegan- expression.2- △ △ CT value is taken to carry out data system
Meter.
1 Real-time RT-PCR primer sequence of table
Fixed right knee joint is taken, formalin is removed, 5% nitric acid solution is substituted for and carries out decalcification.Routine paraffin wax packet
It buries, HE dyeing, carries out histopathologic examination under light microscopic.
The data obtained is counted with 17.0 software of SPSS, and data are usedIt indicates, between measuring result each group
Compare and be all made of variance analysis, P < 0.05 is that difference is statistically significant.
2.2 analgesic activity
2.2.1 mouse hot-plate is tested
KM mouse is selected, female, by it by being only placed on 55 DEG C of metal fever board slots, constant temperature changes in ± 0.5 DEG C, from small
Mouse is put into hot plate slot and starts timing, to lick metapedes as pain reaction indicator.Test advance action object screening, will be less than the response latency
5s or animal greater than 30s are rejected, and are chosen qualified KM mouse 60, are randomly divided into control group；The high agent of zhuanggu guanjie pill
Amount group (3.08g/kg)；Present composition high dose group (3.08g/kg), middle dose group (1.54g/kg), low dose group
(0.77g/kg) 5 dosage groups.It is administered once daily, control group gives corresponding dosage distilled water, is administered 14 days, with hot plate method (55
DEG C ± 0.5 DEG C) pain threshold of measurement mouse, the difference between comparative experiments group and control group.
2.2.2 mouse acetic acid twisting is tested
KM mouse 60 are chosen, half male and half female, grouping is administered and tests with 2.2.1 mouse hot-plate, after last dose 30min,
Only by 0.2mL/, 0.7% ice of intraperitoneal injection acid observes and records 5-10min, 11-15min and 16-20min period after injection
Writhing sum in the writhing number and 15min of interior mouse, the difference between comparative experiments group and control group.
2.3 anti-inflammatory effect
2.3.1 the swollen experiment of mouse dimethylbenzene ear
KM mouse 60 are chosen, female, grouping is administered and tests with 2.2.1 mouse hot-plate, after the last administration 30min, in small
100% dimethylbenzene proinflammatory agent 0.03mL/ of mouse auris dextra tow sides even spread only, with the swollen method of dimethylbenzene ear calculates swelling
(mg), the difference between comparative experiments group and control group.2.3.2 swollen hyperplasia of rat granuloma is tested
Rat 60 are chosen, male is randomly divided into control group；Zhuanggu guanjie pill high dose group (2.20g/kg)；The present invention
Composition high dose group (2.20g/kg), middle dose group (1.10g/kg)；5 dosage groups of low dose group (0.55g/kg).It takes
Swollen hyperplasia of rat granuloma model method rat et al. Ke cotton balls, for 24 hours after, according to dosage set continuous gavage 14d, control group gives
Respective volume distilled water, is administered once daily, and after the last administration, puts to death animal, cotton balls is extracted together with granulation tissue, weigh,
Record weight in wet base.60 DEG C of dryings are weighed afterwards for 24 hours, as dry weight, are subtracted original cotton balls weight, as granulation net weight, are compared administration group
Difference between control group.
3 results
The effect of 3.1 osteoarthritis treatments
3.1.1 to the therapeutic effect of hareley cavy spontaneity osteoarthritis
The cavy of spontaneous osteoarthritis is formed, after successive administration 4 weeks, the weight of each administration group animal is compared with model group
There was no significant difference, MMP-1 and TIMP-1 value is above model group.Histopathological examination shows, model group animal knee joint
Apparent cartilaginous tissue retrogression pathological changes occur, if cartilage four-layer structure is unobvious, cells of superficial layer falls off, a small amount of cell infiltration
And cartilage cell it is downright bad phenomena such as；Each administration group compared with model group, knee cartilage regression be suppressed or delay (be shown in Table 2,
Fig. 1).
The influence of 2 zhuanggu guanjie pill of table and the present composition to guinea pig serum MMP-1, TIMP-1
3.1.2 to the treatment of the rat bone arthritis chronica phase of Papain enzyme induction
As a result, the osteoarthritis chronic phase rat articular liquid MMP-3 content to Papain enzyme induction is high as shown in table 3-1
In Normal group (P < 0.01), the changes of contents of TIMP is unobvious (P > 0.05), and the ratio of MMP-3/TIMP-1 increases,
The content relative increase (P < 0.01) of MMP-3, is controlled with zhuanggu guanjie pill 1.10g/kg and new process 2.20g/kg, 1.10g/kg
It treats 4 weeks, MMP-3 content is substantially reduced (P < 0.01), and the concentration ratio of MMP-3/TIMP-1 is intended to balance.
The influence of table 3-1 zhuanggu guanjie pill and the present composition to joint fluid MMP-3 and TIMP-1
Note: compared with the control group, P < 0.01 * *；Compared with model group, #P < 0.05, ##P < 0.01
As a result, OA chronic phase cartilage of rats Collagen II mRNA and Aggrecan mRNA are bright as shown in table 3-2
It is aobvious to weaken (P < 0.05 or P < 0.01), it is treated 4 weeks with zhuanggu guanjie pill and the present composition, present composition 2.20g/
Kg dosage group rabbit cartilage Collagen II mRNA expression is remarkably reinforced (P < 0.01), zhuanggu guanjie pill 1.10g/kg and sheet
Inventive composition 2.20g/kg, 1.10g/kg dosage group Aggrecan mRNA expression are remarkably reinforced, wherein the present composition
1.10g/kg dosage group and model group difference are statistically significant (P < 0.01).The result shows that zhuanggu guanjie pill and of the present invention group
Closing object can promote the synthesis of Collagen II and Aggrecan, has certain anti-cartilage damage effect, can delay cartilage regression
Property become process.
The influence of table 3-2 zhuanggu guanjie pill and the present composition to cartilage CollagenII, AggrecanmRNA
Note: compared with the control group, * P < 0.05, * * P < 0.01；Compared with model group, #P < 0.05, ##p < 0.01
The discovery of OA rat articular histopathologic examination, gross examination of skeletal muscle: zhuanggu guanjie pill and present composition treatment group:
Rat articular lesion is slightly lighter than model group.PATHOMORPHOLOGICAL OBSERVATION OF PULLORUM: zhuanggu guanjie pill and present composition 2.20g/kg,
1.10g/kg dosage treatment group: compared with model group, cartilaginous tissue necrosis is reduced, and necrotic area remains more cartilage cell, greatly
It is small different, disorganized, some tufted arrangements.
3.2 analgesic activity
3.2.1 hot-plate is shown
30min after medicine, compared with the control group, three dosage groups of the present composition can significantly extend the pain reaction of mouse
Time (p < 0.01) has preferable analgesic activity；60min after medicine, the middle dose group of the only present composition show compared with
Good analgesic effect (p < 0.05)；120min after medicine, each administration group is without analgesic effect (the results are shown in Table 4).
The influence of 4. zhuanggu guanjie pill of table and the present composition to the mouse hot-plate pain reaction time
Note: compared with the control group, examining through t,*P < 0.05,**P < 0.01,***P<0.001
3.2.2 acetic acid twisting is tested
The results show that three dosage groups of the present composition are to mouse writhing after injecting algogen acetic acid to mouse
Number has certain inhibiting effect, and You Yigao, middle dosage analgesic effect are significant (being shown in Table 5).
The influence of 5. zhuanggu guanjie pill of table and the present composition to mouse acetic acid twisting number
Note: compared with the control group,*P < 0.05,**P < 0.01,***P<0.001
3.3 anti-inflammatory effect
The swollen experiment of mouse dimethylbenzene ear and swollen hyperplasia of rat granuloma experiment display, the present composition can inhibit dimethylbenzene institute
Caused Mice Auricle topical acute inflammation and swollen hyperplasia of rat granuloma hyperplasia, be effectively reduced mouse and rat acute inflammation and
The generation (being shown in Table 6,7) of chronic inflammation.
The influence that 6. zhuanggu guanjie pill of table and the present composition swell to mouse caused by dimethylbenzene xylene ear
Note: compared with the control group,*P < 0.05,**P < 0.01,***P<0.001。
The influence of 7. zhuanggu guanjie pill of table and the present composition to swollen hyperplasia of rat granuloma
Note: compared with the control group,*P < 0.05,**P < 0.01,***P<0.001
2 toxicity test of experimental example
1, materials and methods
1.1 test sample
The present composition prepared by embodiment 1, China Resources Sanjiu Medical & Pharmaceutical Co., Ltd., the black water-bindered pill, gas fragrance, taste
Slight bitter；
1.2 reference substance
Zhuanggu guanjie pill, China Resources Sanjiu Medical & Pharmaceutical Co., Ltd., the black water-bindered pill, gas fragrance, mildly bitter flavor.1.3 preparation sides
Method
The present composition and reference substance, according to the weight of measurement the last before every animal drugs, are determined without preparing
The dosage (retain to decimal point 1) of every animal, the amount of weighing of the present composition should control theoretical dosage ±
0.2g.Dosage is calculated by crude drug amount, and 1 gram of new process (preparation of embodiment 1) is equivalent to 1.13 grams of crude drugs, 1 gram of bone strengthening joint
Ball is equivalent to 1 gram of crude drug.
2, experimental system
Beagle dog, regular grade, 60, half male and half female；The animal monthly age: animal was 6~7 monthly ages when administration starts；Animal
Weight: the weight of animals when administration starts: 5.33-6.67kg (♂), 5.44-7.84kg (♂)；Experimental animal source: Beijing Ma Sisheng
Object Technology Co., Ltd.；Experimental animal production licence number: SCXK (capital) 2011-0003；Quality of Experimental Animals quality certification number:
0234473；Production licence signs and issues unit: Science and Technology Commission, Beijing.
3, experimental design
3.1 test groupings and dosage
According to the weight of animals that (D-2) before administration is measured, animal similar in qualified, healthy, weight that selection is quarantined uses meter
Animal is carried out section according to gender and is grouped at random by the random number of calculation machine system.It is dynamic after groups of animals dosage and grouping
Object number see the table below 8:
Each tested group of animal dosage design of table 8
Note:a: human body presses 60kg calculating again, and clinic is intended with dosage being 0.2g/kg/ days (calculating by crude drug amount)
Administration route is oral administration, and administration frequency is weekly administration 6 days, to be administered once a day, successive administration with the period
It 39 weeks, is administered 234 times altogether.
4, living animal Testing index
4.1 general clinical observations
Every morning and afternoon respectively carry out 1 general clinical observation, comprising: cage observation animal dead or dying situation,
The state of mind, behavioral activity, fecal character etc..
The observation of 4.2 Detailed clinicals
Detailed clinical observation is carried out once a week, comprising: the state of mind, behavioral activity, skin, coat, eye, ear, nose, abdomen
Portion, external genital organs, anus, four limbs, mouth, foot and breathing etc..
The observation of 4.3 other parameters
Including irregularly to the weight of animals, body temperature, Ecg.
The monitoring of 4.4 clinical pathologies
Including to blood count, coagulation function (prothrombin time (PT), active partial thromboplastin time
(APTT), fibrinogen (FIB)), blood parameters, uroscopy, eye examination, after animal euthanasia Testing index into
Row detection, while the corpse of the animal after euthanasia is observed, monitors organ weights and organ and group to all animals
It knits and carries out routine paraffin wax embedding, slice, HE dyeing and micro- sem observation.
5, data acquisition and statistical analysis
Data acquisition: scheme requires measurement and observation data result to need written notes to table appropriate or computer
Directly acquire data.
Statistical analysis: all statistical analysis are located at 5% or P≤0.05 using double tail analyses, statistics level.Groups of animals
The indexs such as weight, body temperature, electrocardiogram parameters, blood count, coagulation function, blood biochemical, organ weights and organ coefficient
Calculate the average and standard deviation of different Group Animals (female animals merge).Above-mentioned data will be by following process
Analysis: Homogeneity Test is carried out to data with Levene Test first.If data are uniform (P ﹥ 0.05), single factor test is carried out
Variance analysis；If variance analysis is significant (P≤0.05), it is for statistical analysis to carry out Dunnett ' s.If Levene
The result of Test is significant (P≤0.05), then carries out Kruskal-wallis non-parametric test；If the non-ginseng of Kruskal-wallis
Number inspection result is significant (P≤0.05), then is further examined using Mann-Whitney U for statistical analysis.Sample number (n)
When less than 3, without statistical analysis.
6, result
6.1 animal deads and dying
During test, groups of animals does not occur dead or dying phenomenon.
6.2 clinical observation
During test, the abnormal response of each administration group is mainly the gastrointestinal reaction after medicine, and concrete condition is as follows:
Zhuanggu guanjie pill 3g/kg dosage group: 2 animals vomit after medicine for the first time, and visible other animals go out successively later
It now vomits, visible vomiting after 7/10 animal drugs during administration, it is 9 dogs time, W13 animal that wherein the frequency vomitted, which occurs, in W1 animal
The frequency vomitted occur is 1 dog time, and W27 animal does not vomit, and it is 4 dogs time that the frequency vomitted, which occurs, in W39 animal；1 dynamic
There is loose stools in object (D2) after the 2nd medicine, and soft stool or loose stools occur in visible other animals successively later, during administration 9/10 it is dynamic
Visible soft stool or loose stools after object medicine, it is 2 dogs time that wherein loose stools or the frequency of soft stool, which occurs, in W1 and W13 animal, and W27 and W39 are dynamic
Object does not occur soft stool or loose stools.
Zhuanggu guanjie pill 10g/kg dosage group: 9 animals vomit after medicine for the first time, and visible other animals go out successively later
It now vomits, visible vomiting after 10/10 animal drugs during administration, wherein the frequency vomitted occur in W1, W13, W27 and W39 animal
The dog of respectively 54,25,17 and 12 time；There is loose stools in 1 animal (D2) after the 2nd medicine, and visible other animals go out successively later
Existing soft stool or loose stools, visible soft stool or loose stools after 10/10 animal drugs during administration, wherein W1, W13 and W27 animal occur soft
Just or the frequency of loose stools is respectively 10,13 and 1 dog time, and W39 animal does not occur soft stool or loose stools.
Present composition 1g/kg dosage group: animal does not vomit during administration, soft stool occurs after 1 animal D5 medicine,
There is soft stool or loose stools in visible other animals successively later, visible soft stool or loose stools after 5/10 animal drugs during administration, wherein
It is 1 dog time that soft stool or the frequency of loose stools, which occurs, in W1 animal, and W13, W27 and W39 animal do not occur soft stool or loose stools.
Present composition 3g/kg dosage group: 1 animal vomits after medicine for the first time, visible other dynamic successively later
Object is vomitted, visible vomiting after 4/10 animal drugs during administration, and it is 1 dog time that wherein the frequency vomitted, which occurs, in W1 animal,
W13, W27 and W39 animal do not vomit；There is soft stool after the 2nd medicine of 2 animals, visible other animals occur successively later
Soft stool or loose stools, visible soft stool or loose stools after 5/10 animal drugs during administration, wherein there is the frequency of soft stool or loose stools in W1 animal
Rate is 2 dogs time, and W13, W27 and W39 animal do not occur soft stool or loose stools.
Present composition 10g/kg dosage group: 1 animal vomits after medicine for the first time, visible other dynamic successively later
Object is vomitted, visible vomiting after 10/10 animal drugs during administration, and wherein vomitting occur in W1, W13, W27 and W39 animal
Frequency is respectively 7,2,8 and 2 dogs time；There is soft stool after the 2nd medicine in 2 animals, and visible other animal appearance are soft successively later
Just or loose stools, visible soft stool or loose stools after 10/10 animal drugs during administration, wherein there is soft stool or loose stools in W1 and W13 animal
Frequency be respectively 9 and 6 dogs time, W27 and W39 animal does not occur soft stool or loose stools.
In addition, visible hygrostomia shows during the Some Animals including negative control group are administered, Ke Nengyu
It stress be related；Some Animals visible dermis erythema, flush, anomalies of conjunctiva, discharge of eye, wound, skin ulceration, movement are lost
The performance such as tune is thought of as the changes such as the spontaneous wound for sexually revising or occurring when movable of animal, it is believed that uncorrelated to administration.
6.3 weight
During test, compared with same period negative control group, zhuanggu guanjie pill 10g/kg dosage group animal D49, D56, D63,
The weight loss (P≤0.05) of D70, D77, D84, D91 and D210, the weight of remaining groups of animals there are no toxicological significance
Change；Compared with same period present composition 10g/kg dosage group, zhuanggu guanjie pill 10g/kg dosage group animal D49, D56,
The weight loss (P≤0.05) of D63, D70, D77, D84, D91 and D210.
Different time points groups of animals changes of weight trend is shown in Fig. 2.
6.4 body temperature
During test, the body temperature of all animals has no the change with toxicological significance.
6.5 electrocardiogram
During test, compared with same period negative control group, zhuanggu guanjie pill 10g/kg dosage group and the present composition
The QRS time limit of 10g/kg dosage group animal D272 shortens.Zhuanggu guanjie pill 10g/kg dosage group and new process 10g/kg dosage group
The QRS time limit of animal D272 is compared and has no difference (P > 0.05).
Due to the QRS time limit of zhuanggu guanjie pill 10g/kg dosage group and present composition 10g/kg dosage group animal D272
In reference range, and significant change is also had no compared with before medicine, therefore, it is considered that the variation does not have toxicological significance.
6.6 clinical pathology
6.6.1 blood count
Related indexes of red blood cells
During test, compared with same period negative control group, the Retic of present composition 10g/kg dosage group D273 is (absolutely
To value) it increases, MCH is reduced；The MCH of present composition 3g/kg dosage group D273 is reduced, and the above difference has statistical significance
(P≤0.05)。
Compared with the dosage zhuanggu guanjie pill control group such as same period, the MCH of present composition 3g/kg dosage group animal D273
It reduces (P≤0.05)；The Retic (absolute value) and MCH of present composition 10g/kg dosage group animal D273 has no obvious change
Change (P > 0.05).
Due to the Retic (absolute value) and MCH of present composition 10g/kg dosage group animal D273 and of the present invention group
M- reaction relation when closing the MCH of object 3g/kg dosage group animal D273 in reference range, and having no specific, therefore recognize
Do not have toxicological significance for the variation.
Platelet counts
During test, compared with same period negative control group, the WBC of present composition 10g/kg dosage group animal D184 and
Neut (absolute value) is increased, and the Lymph (absolute value) of D273 is increased；The Mono of zhuanggu guanjie pill 10g/kg dosage group animal D273
(relative value and absolute value) increases, and Eos (relative value) is reduced, and the above difference has statistical significance (P≤0.05).
Compared with the dosage zhuanggu guanjie pill control group such as same period, present composition 10g/kg dosage group animal D184's
WBC and Neut (absolute value) is increased, and the Mono (relative value and absolute value) of D273 is reduced, and EOS (relative value) is increased, the above difference
With statistical significance (P≤0.05).
Due to WBC, Neut (absolute value) of present composition 10g/kg dosage group animal D184, the Lymph of D273 is (absolutely
To value) and zhuanggu guanjie pill 10g/kg dosage group animal D273 Mono (relative value and absolute value) and Eos (relative value)
In reference range, and while having no specific m- reaction relation, therefore, it is considered that the variation does not have toxicological significance.
Blood platelet
During test, compared with same period negative control group, zhuanggu guanjie pill 3g/kg dosage group animal D91, D184 and D273
PLT increase, the PLT of zhuanggu guanjie pill 10g/kg dosage group animal D29, D91, D184 and D273 is increased；The present composition
The PLT of 3g/kg and 10g/kg dosage group animal D29, D91, D184 and D273 is increased, the above difference have statistical significance (P≤
0.05)。
Compared with the dosage present composition group such as same period, the PLT of zhuanggu guanjie pill 0g/kg group animal D273 increase (P≤
0.05)。
At the end of convalescence, the regularity that each administration group blood platelet is shown no obvious abnormalities changes.
Groups of animals PLT situation of change see the table below 9 during administration:
Groups of animals PLT situation of change during table 9 is administered
Note: 1. " * " indicate P≤0.05 compared with vehicle control group；2. " ↑ ", indicates to increase
The situation of change of above-mentioned PLT prompts: after administration, zhuanggu guanjie pill and the present composition 3,10g/kg dosage group are dynamic
Significantly raised phenomenon is presented in object PLT, and the above phenomenon thinks related to administration.6.6.2 coagulation function
During test, each administration group animal Blood Coagulation there are no the change of toxicological significance.
6.6.3 blood biochemical
During test, with same period negative control most compared with, the ALT and ALB of zhuanggu guanjie pill 3g/kg dosage group animal D91
It reduces, ALP is increased, and the ALP of D184 is increased, and the ALP and TCH of D273 is increased；
ALT, ALB and A/G of zhuanggu guanjie pill 10g/kg dosage group animal D29 declines, and TG is increased, ALT, AST of D91,
ALB, A/G and Cl- are reduced, and ALP and TG are increased, and A/G, BUN, CREA and Cl- of D184 is reduced, and ALP, GLB and K+ are increased, D273
AST, ALB, A/G, BUN, CREA, Na+ and Cl- reduce, ALP, GGT and TG increase；
The ALB and A/G of the present composition (preparation of embodiment 1) 1g/kg dosage group animal D91 and D184 are reduced；
The A/G of present composition 3g/kg dosage group animal D29 is reduced, and the ALB of D91 is reduced, the TCH of D184 and D273
It increases；
The ALT and A/G of present composition 10g/kg dosage group animal D29 are reduced, and GLB is increased, ALT, ALB of D91 and
A/G is reduced, and the ALB and A/G of D184 is reduced, and GLB and TCH are increased, and the ALB and A/G of D273 is reduced, and TCH is increased, the above difference tool
Statistically significant (P≤0.05).Remaining blood parameters has no notable difference.
Compared with the dosage present composition such as same period (preparation of embodiment 1) group, zhuanggu guanjie pill 3g/kg dosage group animal
The ALP of D91, D184 and D273 are increased, and the TCH of D91 and D184 are reduced；The TG of zhuanggu guanjie pill 10g/kg dosage group animal D29
It increasing, the CREA and TCH of D91, D184 and D273 are reduced, and the Cl- of D91 and D273 are reduced, and the ALP of D184 and D273 are increased,
AST, BUN, A/G and Na+ of D273 is reduced, and GGT is increased, and the above difference has statistical significance (P≤0.05).
In the blood parameters that above-mentioned appearance changes, due to ALT, AST, BUN, CREA, Na+、K+And Cl-Referring to
It is worth fluctuation in range, has no that evident regularity changes, therefore, it is considered that uncorrelated to administration.Remaining blood parameters situation of change
Such as the following table 10.
10 blood parameters situation of change of table
Note: " ↓ " indicates to reduce；" ↑ ", indicates to increase；And compared with same period negative control group, the statistically significant (P of difference
≤0.05)
In the data of above-mentioned variation, groups of animals before medicine compared with being worth such as the following table 11:
It is worth before 11 groups of animals of table and medicine and compares
Note: " ↓ " indicates to reduce；" ↑ ", indicates to increase
Blood parameters situation of change prompts in upper table: during administration, zhuanggu guanjie pill 3g/kg and 10g/kg dosage group
Animal ALP is significantly raised, and visible GGT is significantly increased at the end of zhuanggu guanjie pill 10g/kg dosage group animal is administered, it is believed that ALP and
The raising of GGT and the hepatotoxicity wind agitation of zhuanggu guanjie pill are related, though the visible ALP liter of present composition 10g/kg dosage group animal D184
Height, but significant change is had no compared with before medicine；Zhuanggu guanjie pill 3g/kg and 10g/kg and the present composition 3g/kg and 10/kg
The visible TCH of dosage group animal is increased, and the visible TG of zhuanggu guanjie pill 10g/kg dosage group animal is increased, it is believed that the variation and administration phase
It closes；The visible ALB of each administration group is slightly reduced, and zhuanggu guanjie pill 10g/kg and the visible A/G of each dosage group of the present composition are reduced,
Zhuanggu guanjie pill and the visible GLB of present composition 10g/kg group are increased, in conjunction with combination pathological examination result, it is believed that the variation
May be related to administration, but each dosage group ALB of the present composition has no significant change, the slight drop of A/G compared with before administration
It is low may be related with the slightly raising of GLP after medicine.
6.7 urinalysis
During test, each administration group animals urine inspection there are no the change of toxicological significance.
6.8 eye examination
During test, each administration group animal eye examination there are no the change of toxicological significance.
6.9 pathologic finding
Compared with same period negative control group, at the end of the administration phase, zhuanggu guanjie pill group 3g/kg dosage group animal's liver weight
It increases, than increasing, epididymis weight is reduced for zhuanggu guanjie pill 10g/kg dosage group animal's liver weight and dirty body, and the above difference has
Statistical significance (P≤0.05).
Compared with the dosage present composition such as same period (preparation of embodiment 1) group, zhuanggu guanjie pill 3g/kg dosage group animal
Liver weight increases, and than increasing, epididymis weight reduces the dirty body of zhuanggu guanjie pill 10g/kg dosage group animal's liver, the above difference tool
Statistically significant (P≤0.05).
Histological indications show that the toxic pathology of the visible liver of zhuanggu guanjie pill 10g/kg dosage group animal changes,
Therefore the weight change of zhuanggu guanjie pill control group liver may be related to administration in this test.
6.10 gross examination of skeletal muscle
All animal gross examination of skeletal muscle are showed no obvious exception relevant to drug.
6.11 histological examination
Microscopic examination showed, the visible Histological change relevant to zhuanggu guanjie pill of liver.Successive administration 39 weeks, bone strengthening
Arthrosis pill 10g/kg group has the visible Binucleate Hepatocytes of 2/6 animal's liver to increase, the visible bile duct proliferation of 1/6 animal, and 1/6 animal is visible
Degeneration of liver cells restores completely after restoring 4 weeks；Zhuanggu guanjie pill 3g/kg group has the visible bile duct of 1/4 animal at the end of restoring 4 weeks
Hyperplasia.
And the present composition (preparation of embodiment 1) each dosage group has no obviously Histological change relevant to new process.
It discusses
The analysis of 7.1 toxic reactions
During administration, the stomaches such as present composition group and the main visible vomiting of bone strengthening arthritis control group, soft stool or loose stools
Enteron aisle reaction, and the weight loss that the visible gastrointestinal reaction of bone strengthening arthritis 10g/kg group is secondary.
Bone strengthening arthritis control group 3g/kg and 10g/kg dosage group animal ALP is significantly raised, bone strengthening arthritis control group
Visible GGT is significantly increased at the end of 10g/kg dosage group animal is administered, it is believed that the raising of ALP and GGT and the arthritic liver of bone strengthening
Toxicity is related.
The bone strengthening arthritis control group 3g/kg and 10g/kg and present composition (preparation of embodiment 1) 3g/kg and 10g/kg
The visible TCH of dosage group animal is increased, and the visible TG of bone strengthening arthritis control group 10g/kg dosage group animal is increased；Bone strengthening arthritis pair
It is reduced according to the visible ALB of each dosage group is organized slightly, bone strengthening arthritis control group 10g/kg and the present composition (preparation of embodiment 1)
The visible A/G of each dosage group is reduced, bone strengthening arthritis and the visible GLB raising of present composition 10g/kg group, it is believed that the variation can
Can be related to administration, but the slight decrease of each dosage group A/G of the present composition may be related with the slightly raising of GLB.
7.2 toxic reactions compare
During administration, the gastrointestinal tracts such as the present composition and the visible vomiting of bone strengthening arthritis control group, soft stool or loose stools
Stimulate the reaction, but under Isodose, occur in the frequency and the extent of reaction that GI irritation is reacted, the arthritic stomach and intestine of bone strengthening
Road reaction is slightly heavy, and secondary weight loss performance occurs in bone strengthening arthritis；And the present composition 3 and 10g/kg and bone strengthening are closed
The visible PLT of the scorching control 3 and 10g/kg dosage group of section is increased, but under Isodose, the raised amplitude of present composition PLT
Slightly below bone strengthening arthritis.It is significantly raised that bone strengthening arthritis compares the visible ALP of 3 and 10g/kg group, and bone strengthening arthritis compares
GGT at the end of 10g/kg group is administered is increased；The ALP of present composition 10g/kg group D184 is increased, but amplitude of variation is more same
The control of the dosage bone strengthening arthritis such as phase is small；The visible liver weight of bone strengthening arthritis 3g/kg and 10g/kg dosage group increases, and liver can
See that Binucleate Hepatocytes increase, reaction, each dosage group liver of the present composition such as bile duct proliferation and degeneration of liver cells have no obvious
Change.
The toxicity comparison of equal dosage zhuanggu guanjie pill and the present composition is listed as follows table 12-14.
The comparison of the gastrointestinal toxicity of the dosage zhuanggu guanjie pills such as table 12 and the present composition
The comparison of the hepatotoxicity wind agitation of the dosage zhuanggu guanjie pills such as table 13 and the present composition
The variation of the index of the dosage zhuanggu guanjie pills such as table 14 and the present composition
Note: " ↑ " indicates to increase；And compared with same period negative control group, difference is statistically significant (P≤0.05)
8 conclusions
Under this experimental condition, using zhuanggu guanjie pill control and the embodiment of the present invention 1 prepare pill repeat take orally to
Beagle dog is given, weekly administration 6 days, is administered once a day, successive administration 39 weeks, zhuanggu guanjie pill dosage was 3 and 10g/ respectively
Kg, major toxicity reaction are gastrointestinal reaction (predominantly vomiting, soft stool or loose stools) and hepatic injury；It is prepared by the embodiment of the present invention 1
Pill be 1,3 and 10g/kg respectively, major toxicity reaction is gastrointestinal reaction, the relatively equal dosage zhuanggu guanjie pill of gastrointestinal reaction
(original process) gently, hepatotoxicity wind agitation damage is lower.
Detailed description of the invention
In order to make the content of the present invention more clearly understood, it below according to specific embodiments of the present invention and combines
Attached drawing, the present invention is described in further detail, wherein
Fig. 1 is that the H.E of osteoarthritis rat knee joints cartilaginous tissue effect of each test group to Papain enzyme induction contaminates
Chromatic graph piece (× 200)；Wherein, A is control group, B is model group, C is zhuanggu guanjie pill group, D-F is respectively 1 test drug of embodiment
Object 2.20,1.10,0.55g/kg group, G are Glucosamine control group；
Fig. 2 is different time points groups of animals changes of weight trend in toxicity test.
Specific embodiment
Embodiment 1
576.8 grams of [prescription] rhizoma cibotii, 346.1 grams of Herba Epimedii, 346.1 grams of Radix Angelicae Pubescentis, 462.0 grams of the rhizome of davallia, teasel root 576.8
Gram, 576.8 grams of herba taxilli, 346.1 grams of Caulis Spatholobi, 1384.2 grams of Rehmannia glutinosa, 346.1 grams of radix aucklandiae, 346.1 grams of olibanum, myrrh
346.1 grams, 346.1 grams of psoralea corylifolia.
[preparation method]
It takes the olibanum and myrrh to mix according to selected weight, adds 10 times of amount water to carry out steam distillation extraction 12 small
When, collect volatile oil；According to volatile oil: beta-cyclodextrin: water: auxiliary material is added in the ratio that the mass ratio of ethyl alcohol is 1:10:20:1, grinds
Mill 20min is included, and dry within 50 DEG C, crushing crosses 120 meshes, volatile oil clathrate compound is made, spare；
The psoralea corylifolia is taken according to selected weight, adds 8 times of amount water to extract 2 times, 1 hour every time, is received after discarding Aqueous extracts
Collect the dregs of a decoction, is drying to obtain psoralea corylifolia processed product, it is spare；
Remaining medicinal material is taken according to selected weight, is mixed with psoralea corylifolia processed product, crushes, sieving, inclusion essential oil is added
Object and suitable adhesive mix, dry with water pill, film coating to get.
Embodiment 2
290 grams of [prescription] rhizoma cibotii, 520 grams of Herba Epimedii, 154 grams of Radix Angelicae Pubescentis, 692 grams of the rhizome of davallia, 290 grams of teasel root, herba taxilli 864
Gram, 154 grams of Caulis Spatholobi, 2076 grams of Rehmannia glutinosa, 154 grams of radix aucklandiae, 520 grams of olibanum, 154 grams of myrrh and 520 grams of psoralea corylifolia.
[preparation method]
It takes the olibanum and myrrh to mix according to selected weight, adds 8 times of amount water to carry out steam distillation and extract 15 hours,
Collect volatile oil；According to volatile oil: beta-cyclodextrin: water: auxiliary material, grinding is added in the ratio that the mass ratio of ethyl alcohol is 1:8:24:1
20min is included, and volatile oil clathrate compound is made in dry within 50 DEG C, crushing, spare；
The psoralea corylifolia is taken according to selected weight, 6 times of amount water is added to extract 3 times, 0.5 hour every time, after discarding Aqueous extracts
The dregs of a decoction are collected, it is spare to be drying to obtain psoralea corylifolia processed product；
Remaining medicinal material is taken according to selected weight, is mixed with psoralea corylifolia processed product, crushes, sieving, inclusion essential oil is added
Object and suitable adhesive mix, dry with water pill, film coating to get.
Embodiment 3
864 grams of [prescription] rhizoma cibotii, 154 grams of Herba Epimedii, 520 grams of Radix Angelicae Pubescentis, 224 grams of the rhizome of davallia, 864 grams of teasel root, herba taxilli 290
Gram, 520 grams of Caulis Spatholobi, 692 grams of Rehmannia glutinosa, 520 grams of radix aucklandiae, 154 grams of olibanum, 520 grams of myrrh and 154 grams of psoralea corylifolia.
[preparation method]
It takes the olibanum and myrrh to mix according to selected weight, adds 12 times of amount water to carry out steam distillation and extract 8 hours,
Collect volatile oil；According to volatile oil: beta-cyclodextrin: water: auxiliary material, grinding is added in the ratio that the mass ratio of ethyl alcohol is 1:12:16:2
20min is included, dry within 50 DEG C, crushing, volatile oil clathrate compound is made, it is spare；
The psoralea corylifolia is taken according to selected weight, adds 10 times of amount water to extract 1 time, 0.5 hour, is collected after discarding Aqueous extracts
It is spare to be drying to obtain psoralea corylifolia processed product for the dregs of a decoction；
Remaining medicinal material is taken according to selected weight, is mixed with psoralea corylifolia processed product, crushes, sieving, inclusion essential oil is added
Object and suitable adhesive mix, dry with water pill, film coating to get.
Embodiment 4
576.8 grams of [prescription] rhizoma cibotii, 346.1 grams of Herba Epimedii, 346.1 grams of Radix Angelicae Pubescentis, 462.0 grams of the rhizome of davallia, teasel root 576.8
Gram, 576.8 grams of herba taxilli, 346.1 grams of Caulis Spatholobi, 1384.2 grams of Rehmannia glutinosa, 346.1 grams of radix aucklandiae, 346.1 grams of olibanum, myrrh
346.1 grams, 346.1 grams of psoralea corylifolia.
[preparation method]
It takes the olibanum and myrrh to mix according to selected weight, adds 10 times of amount water to carry out steam distillation extraction 12 small
When, volatile oil is collected, customary adjuvant is added, volatile oil clathrate compound is made, it is spare；
The psoralea corylifolia is taken according to selected weight, adds 8 times of amount water to extract 2 times, 1 hour every time, is received after discarding Aqueous extracts
Collect the dregs of a decoction, it is spare to be drying to obtain psoralea corylifolia processed product；
Remaining medicinal material is taken according to selected weight, is mixed with the volatile oil clathrate compound and psoralea corylifolia processed product, is crushed, mistake
Sieve, is added suitable customary adjuvant, honeyed bolus is made.
Embodiment 5
576.8 grams of [prescription] rhizoma cibotii, 346.1 grams of Herba Epimedii, 346.1 grams of Radix Angelicae Pubescentis, 462.0 grams of the rhizome of davallia, teasel root 576.8
Gram, 576.8 grams of herba taxilli, 346.1 grams of Caulis Spatholobi, 1384.2 grams of Rehmannia glutinosa, 346.1 grams of radix aucklandiae, 346.1 grams of olibanum, myrrh
346.1 grams, 346.1 grams of psoralea corylifolia.
[preparation method]
It takes the olibanum and myrrh to mix according to selected weight, adds water to carry out steam distillation extraction, collect volatile oil,
Customary adjuvant is added, volatile oil clathrate compound is made, it is spare；
The psoralea corylifolia is taken according to selected weight, extracting in water collects the dregs of a decoction after discarding Aqueous extracts, is drying to obtain Psoralen
Rouge processed product, it is spare；
Remaining medicinal material is taken according to selected weight, is mixed with the psoralea corylifolia processed product, crushes, sieving, volatile oil is added
Inclusion compound and suitable customary adjuvant, are made tablet.
Obviously, the above embodiments are merely examples for clarifying the description, and does not limit the embodiments.It is right
For those of ordinary skill in the art, can also make on the basis of the above description it is other it is various forms of variation or
It changes.There is no necessity and possibility to exhaust all the enbodiments.And it is extended from this it is obvious variation or
It changes still within the protection scope of the invention.
Claims (8)
1. a kind of preparation method for the pharmaceutical composition for treating osteoarthropathy, which comprises the steps of:
(1) 346.1 parts by weight of olibanum, the mixing of 346.1 parts by weight of myrrh are taken, adds water to carry out steam distillation extraction, collects volatilization
Oil is added customary adjuvant and volatile oil clathrate compound is made, spare；
(2) 346.1 parts by weight of psoralea corylifolia are taken, extracting in water collects the dregs of a decoction after discarding Aqueous extracts, obtains psoralea corylifolia processed product, standby
With；
(3) take 576.8 parts by weight of rhizoma cibotii, 346.1 parts by weight of Herba Epimedii, 346.1 parts by weight of Radix Angelicae Pubescentis, 462.0 parts by weight of the rhizome of davallia,
576.8 parts by weight of teasel root, 576.8 parts by weight of herba taxilli, 346.1 parts by weight of Caulis Spatholobi, 1384.2 parts by weight of Rehmannia glutinosa, radix aucklandiae
346.1 parts by weight are mixed with the spare psoralea corylifolia processed product, are crushed, and add the spare volatile oil clathrate compound and often
Auxiliary material is advised, clinically-acceptable oral preparation is made according to common process.
2. the preparation method of pharmaceutical composition according to claim 1, which comprises the steps of:
(1) it takes the olibanum and myrrh to mix according to selected parts by weight, adds 8-12 times to measure water and carry out steam distillation extraction 8-
15 hours, volatile oil is collected, customary adjuvant is added, volatile oil clathrate compound is made, it is spare；
(2) psoralea corylifolia is taken according to selected parts by weight, adds 6-10 times to measure water and extracts 1-3 times, it is 0.5-2 hours each, it discards
The dregs of a decoction are collected after Aqueous extracts, are drying to obtain psoralea corylifolia processed product, it is spare；
(3) rhizoma cibotii, Herba Epimedii, Radix Angelicae Pubescentis, the rhizome of davallia, teasel root, herba taxilli, Caulis Spatholobi, Rehmannia glutinosa, wood are taken according to selected parts by weight
Perfume (or spice) is mixed with the spare psoralea corylifolia processed product, is crushed, and is added the spare volatile oil clathrate compound and customary adjuvant, is pressed
Clinically-acceptable oral preparation is made in more solito technique.
3. the preparation method of pharmaceutical composition according to claim 2, which comprises the steps of:
(1) it takes the olibanum and myrrh to mix according to selected parts by weight, adds 10 times of amount water to carry out steam distillation extraction 12 small
When, volatile oil is collected, customary adjuvant is added, volatile oil clathrate compound is made, it is spare；
(2) psoralea corylifolia is taken according to selected parts by weight, adds 8 times of amount water to extract 2 times, 1 hour every time, is received after discarding Aqueous extracts
Collect the dregs of a decoction, is drying to obtain psoralea corylifolia processed product, it is spare；
(3) rhizoma cibotii, Herba Epimedii, Radix Angelicae Pubescentis, the rhizome of davallia, teasel root, herba taxilli, Caulis Spatholobi, Rehmannia glutinosa, wood are taken according to selected parts by weight
Perfume (or spice) is mixed with the spare psoralea corylifolia processed product, is crushed, and is added the spare volatile oil clathrate compound and customary adjuvant, is pressed
Clinically-acceptable oral preparation is made in more solito technique.
4. the preparation method of pharmaceutical composition according to claim 1 to 3, which is characterized in that in the step (1),
The customary adjuvant for preparing the volatile oil clathrate compound includes beta-cyclodextrin, water and ethyl alcohol.
5. the preparation method of pharmaceutical composition according to claim 4, which is characterized in that described to wave in the step (1)
Hair oil, beta-cyclodextrin, water and ethyl alcohol mass ratio be 1:8-12:16-24:1-2.
6. the preparation method of pharmaceutical composition according to claim 5, which is characterized in that described to wave in the step (1)
Hair oil, beta-cyclodextrin, water and ethyl alcohol mass ratio be 1:10:20:1.
7. the preparation method of pharmaceutical composition according to claim 5 or 6, which is characterized in that the olibanum and/or myrrh
For vinegar toast.
8. the pharmaceutical composition being prepared by any preparation method of claim 2-7.
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| CN104435314B (en) * | 2014-11-14 | 2018-02-27 | 华润三九医药股份有限公司 | A kind of pharmaceutical composition for treating osteoarthropathy and its application |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1931277A (en) * | 2005-10-08 | 2007-03-21 | 周小明 | Osteoarthrosis treating Chinese medicine composition and its prepn process |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1931277A (en) * | 2005-10-08 | 2007-03-21 | 周小明 | Osteoarthrosis treating Chinese medicine composition and its prepn process |
Non-Patent Citations (3)
| Title |
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| 乳香没药挥发油入药的β-环糊精包合工艺及镇痛作用研究;谈英等;《中国药物警戒》;20100731;第7卷(第7期);390-391 |
| 壮骨关节丸中肝毒性药材的筛选研究;周昆等;《中国药物警戒》;20091130;第6卷(第11期);641-648 |
| 补骨脂水提取引起的大鼠肝损害;周昆等;《天津中医药大学学报》;20131231;第32卷(第4期);221-224 |
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