Summary of the invention
The pharmaceutical composition containing Artemether and tripterygium hypoglaucum hutch that the purpose of the present invention is to provide a kind of, the composition utilize Hao Jia
The synergistic function of ether and two kinds of ingredients of tripterygium hypoglaucum hutch, improves to arthritic curative effect, shortens treatment time, reduces simultaneously
Toxicity.
In order to achieve the goal above, the present invention provides following technical schemes:
A kind of pharmaceutical composition containing Artemether and tripterygium hypoglaucum hutch, including Artemether and tripterygium hypoglaucum hutch, the mass ratio of the two are 0.1
~0.2:7.5~15.
It is a discovery of the invention that can improve to arthritic curative effect after Artemether is compounded with tripterygium hypoglaucum hutch, shorten treatment time,
Reduce toxicity.
It is found using rat, mouse as subjects, pharmaceutical composition of the invention can significantly inhibit caused by mouse red blood cell
Mouse hemolytic antibody generate, moreover it is possible to significantly inhibit mouse delayed hypersensitive reaction caused by 2,4-dinitrofluorobenzene, can be significant
Reduce the Articular swelling of rat.
It tests and finds through mouse toxicity, under same large dosage, Artemether Hao Jia single compared with after tripterygium hypoglaucum hutch compounding
Ether toxicity lethality reduces 6%, reduces 12% compared to single tripterygium hypoglaucum hutch toxicity lethality.
In the present invention, the arbitrary value in the range of 0.1~0.2:7.5~15 is can be used in the mass ratio of Artemether and tripterygium hypoglaucum hutch,
Wherein, when preferably the mass ratio of Artemether and tripterygium hypoglaucum hutch is 0.1:7.5~15, cooperative synergism effect is more significant.
Tripterygium hypoglaucum hutch of the present invention is the root of tripterygium hypoglaucum hutch, directly extract obtained by, can be used conventional extracting method and
, such as extraction etc., i.e., tripterygium hypoglaucum hutch of the present invention are torch flower extract.
Artemether of the present invention can be made by oneself or commercially available.
In some embodiments, pharmaceutical composition further includes pharmaceutically acceptable at least one adjuvant.
In some embodiments, the adjuvant includes solubilizer, preservative, corrigent, wetting agent, disintegrating agent, helps
One of suspension, chelating agent, filler, lubricant, binder, glidant are a variety of.
Adjuvant is selected generally according to preparation type.
Arbitrary preparation, such as tablet, oral solution, powder, capsule, injection is made in pharmaceutical composition of the present invention
Agent or externally dressing.Different dosage forms select different adjuvants.
By taking tablet as an example, the filler is selected from one of lactose, dextrin, mannitol and sorbierite or a variety of, is making
Content in agent is preferably 10~30wt%;
The binder is pregelatinized starch, and content in the formulation is preferably 2~10wt%;
The disintegrating agent is crospovidone, and content in the formulation is preferably 0.5~5wt%;
The lubricant is lauryl sodium sulfate, and content in the formulation is preferably 1~5wt%;
The glidant is talcum powder, and content in the formulation is preferably 1~5wt%.
By taking oral solution as an example, common adjuvant has solubilizer, preservative, corrigent, suspending agent, wetting agent, surface living
Property agent, emulsifier etc..
Pharmaceutical composition of the present invention the preparation method comprises the following steps: according to formula, by all material combinations.
The preparation method of preparation of the present invention is using conventional preparation method, by taking tablet as an example, by all originals
Expect that granulation, tabletting after mixing to obtain the final product.Suitable prilling process can be selected according to the type of auxiliary agent and the physico-chemical property of drug.
To sum up, compared with prior art, invention achieves following technical effects:
(1) after Artemether is compounded with tripterygium hypoglaucum hutch, synergistic function is generated, improves to arthritic curative effect, shortens
Treatment time, while reducing toxicity.
(2) tablet hurtless measure provided by, convenient drug administration, compliance are good.
Specific embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person carries out according to conventional conditions or manufacturer's recommended conditions.Reagents or instruments used without specified manufacturer is
The conventional products that can be obtained by commercially available purchase.
Effect experiment
1, dose design
Artemether dosage: mouse: A1 200mg/kg, A2 100mg/kg;Rat: A3 100mg/kg, A4 50mg/
kg;
Tripterygium hypoglaucum hutch dosage: mouse: B1 is 15g crude drug/kg, and B2 is 7.5g crude drug/kg;Rat: B3 is 7.5g crude drug/kg,
B4 is 3.75g crude drug/kg.
2 experiment contents
The influence that 2.1 pairs of mouse hemolytic antibodies generate
2.1.1 drug is prepared
Test group medication: taking 200mg Artemether, is added after 0.5%CMC-Na solution is sufficiently milled uniformly and is settled to 20ml,
The Artemether solution for being 10mg/ml up to concentration;1.1g colguhoumia root medicinal extract (content is 13.64g crude drug/g medicinal extract) is taken, is added
0.5%CMC-Na solution is settled to 20ml to get concentration after sufficiently milling uniformly be 0.75g crude drug/ml colguhoumia root medicinal extract
Solution.By prepared concentration be 10mg/ml Artemether solution and concentration be that 0.75g crude drug/ml colguhoumia root medicinal extract is molten
To get the solution of required proportion (A2:B2), (the final concentration of 5mg/ml of Artemether, colguhoumia root are whole after liquid 1:1 mixing by volume
Concentration is 0.375g crude drug/ml).
Being settled to 20ml after taking 200mg Artemether, addition 0.5%CMC-Na solution sufficiently to mill uniformly to get concentration is
The Artemether solution of 10mg/ml;2.2g colguhoumia root medicinal extract (content is 13.64g crude drug/g medicinal extract) is taken, 0.5%CMC- is added
Na solution is settled to 20ml to get concentration after sufficiently milling uniformly be 1.5g crude drug/ml colguhoumia root medicinal extract solution.It will prepare
The Artemether solution and concentration that good concentration is 10mg/ml are 1.5g crude drug/ml colguhoumia root medicinal extract solution 1:1 by volume
To get the solution of required proportion (A2:B1), (the final concentration of 5mg/ml of Artemether, the final concentration of 0.75g of colguhoumia root are raw after mixing
Medicine/ml).
Being settled to 20ml after taking 400mg Artemether, addition 0.5%CMC-Na solution sufficiently to mill uniformly to get concentration is
The Artemether solution of 20mg/ml;It takes 1.1g colguhoumia root medicinal extract (13.64g crude drug/g medicinal extract), 0.5%CMC-Na solution is added
It is 0.75g crude drug/ml colguhoumia root medicinal extract solution that 20ml is settled to after sufficiently milling uniformly to get concentration.It will be prepared
The Artemether solution and concentration that concentration is 20mg/ml are that 1:1 is mixed by volume for 0.75g crude drug/ml colguhoumia root medicinal extract solution
To get the solution of required proportion (A1:B2), (the final concentration of 10mg/ml of Artemether, the final concentration of 0.375g of colguhoumia root are raw after conjunction
Medicine/ml).
Being settled to 20ml after taking 400mg Artemether, addition 0.5%CMC-Na solution sufficiently to mill uniformly to get concentration is
The Artemether solution of 20mg/ml;2.2g colguhoumia root medicinal extract (content is 13.64g crude drug/g medicinal extract) is taken, 0.5%CMC- is added
Na solution is settled to 20ml to get concentration after sufficiently milling uniformly be 1.5g crude drug/ml colguhoumia root medicinal extract solution.It will prepare
The Artemether solution and concentration that good concentration is 20mg/ml are 1.5g crude drug/ml colguhoumia root medicinal extract solution 1:1 by volume
To get solution (the final concentration of 10mg/ml of Artemether, the final concentration of 0.75g of colguhoumia root of required proportion (A1:B1) after mixing
Crude drug/ml).
200mg Artemether is taken, is added after 0.5%CMC-Na solution is sufficiently milled uniformly and is settled to 20ml to get required
Artemether solution (A1) (the final concentration of 10mg/ml of Artemether).
1.1g colguhoumia root medicinal extract (content is 13.64g crude drug/g medicinal extract) is taken, 0.5%CMC-Na solution is added and sufficiently grinds
Mill uniformly after be settled to 20ml to get required colguhoumia root medicinal extract solution (B1) (the final concentration of 0.75g crude drug of colguhoumia root/
ml)。
2.1.2 experimental method
Under disinfecting action, mouse blood is added in 0.9% sodium chloride injection of 50mL by the venous blood collection under mouse
It mixes, is then washed 3 times with 0.9% sodium chloride injection, preceding centrifugal speed twice is 1500r/min, is centrifuged 5min, abandons supernatant
The leukocytic cream of liquid and interface is finally centrifuged (2000r/min, 5min) twice in succession again until hematocrit value is constant, by this
Value is made into 5% mouse red blood cell suspension with 0.9% sodium chloride injection.
Culling heart blood is carried out to mouse, after waiting for blood to solidify, 2000r/min is centrifuged 5min, the serum mixing of top is drawn,
10% serum is made into physiological saline.
After quarantine and adaptive feeding, choose 56 qualified mouse of health, weight be no more than average weight ±
20%, it is randomly divided into 7 groups, i.e. blank control group, Artemether and tripterygium hypoglaucum hutch A2:B2 group, Artemether and tripterygium hypoglaucum hutch A1:B2 group, wormwood artemisia
Methyl ether and tripterygium hypoglaucum hutch A2:B1 group, Artemether and tripterygium hypoglaucum hutch A1:B1 group, Artemether A1 group, tripterygium hypoglaucum hutch B1 group, every group of 8 (male and female
It is fifty-fifty).After grouping, corresponding drug is given in stomach-filling to each medicine group respectively, and blank control group gives the purified water of respective volume,
Administered volume is 0.2ml/10g weight, once a day, successive administration 7 times.
Administration first day, 5% mouse red blood cell suspension is only injected intraperitoneally by 0.2ml/ and is immunized for each group.
1h after the last administration, eyeball take blood, and 3000r/min is centrifuged 10min, serum are separated, with 0.9% sodium chloride injection
Serum is diluted 100 times.Dilute serum 1ml is taken, with 5% mouse red blood cell suspension 0.5ml, 10% mice serum (complement)
0.5ml is mixed, and is incubated 30min in 37 DEG C of insulating boxs, 30min stopped reaction in 0 DEG C of ice water is then set, by aforesaid liquid 3000r/
Min is centrifuged 10min, takes supernatant (the supernatant zeroing that mice serum is not added), measures OD at 540nm with spectrophotometer
Value.
2.1.3 experimental result
Such as table 1, the different ratio of Artemether and tripterygium hypoglaucum hutch generates mouse hemolytic antibody caused by mouse red blood cell equal
There is apparent inhibiting effect, it is prompted to have inhibiting effect to humoral immune function, and is substantially better than the drug being applied alone.
Table 1
Group |
Ratio |
OD value |
Blank control |
/ |
1.260±0.709 |
Artemether: tripterygium hypoglaucum hutch |
A2:B2 |
0.091±0.099 |
Artemether: tripterygium hypoglaucum hutch |
A1:B2 |
0.140±0.187 |
Artemether: tripterygium hypoglaucum hutch |
A2:B1 |
0.103±0.150 |
Artemether: tripterygium hypoglaucum hutch |
A1:B1 |
0.178±0.183 |
Artemether |
A1 |
0.191±0.317 |
Tripterygium hypoglaucum hutch |
B1 |
0.149±0.255 |
2.2 couples of DNFB cause the influence of mouse delayed hypersensitive reaction
2.2.1 drug is prepared
Test group medication: taking 200mg Artemether, is added after 0.5%CMC-Na solution is sufficiently milled uniformly and is settled to 20ml,
The Artemether solution for being 10mg/ml up to concentration;1.1g colguhoumia root medicinal extract (content is 13.64g crude drug/g medicinal extract) is taken, is added
0.5%CMC-Na solution is settled to 20ml to get concentration after sufficiently milling uniformly be 0.75g crude drug/ml colguhoumia root medicinal extract
Solution.By prepared concentration be 10mg/ml Artemether solution and concentration be that 0.75g crude drug/ml colguhoumia root medicinal extract is molten
To get the solution of required proportion (A2:B2), (the final concentration of 5mg/ml of Artemether, colguhoumia root are whole after liquid 1:1 mixing by volume
Concentration is 0.375g crude drug/ml).
Being settled to 20ml after taking 200mg Artemether, addition 0.5%CMC-Na solution sufficiently to mill uniformly to get concentration is
The Artemether solution of 10mg/ml;2.2g colguhoumia root medicinal extract (content is 13.64g crude drug/g medicinal extract) is taken, 0.5%CMC- is added
Na solution is settled to 20ml to get concentration after sufficiently milling uniformly be 1.5g crude drug/ml colguhoumia root medicinal extract solution.It will prepare
The Artemether solution and concentration that good concentration is 10mg/ml are 1.5g crude drug/ml colguhoumia root medicinal extract solution 1:1 by volume
To get the solution of required proportion (A2:B1), (the final concentration of 5mg/ml of Artemether, the final concentration of 0.75g of colguhoumia root are raw after mixing
Medicine/ml).
Being settled to 20ml after taking 400mg Artemether, addition 0.5%CMC-Na solution sufficiently to mill uniformly to get concentration is
The Artemether solution of 20mg/ml;It takes 1.1g colguhoumia root medicinal extract (13.64g crude drug/g medicinal extract), 0.5%CMC-Na solution is added
It is 0.75g crude drug/ml colguhoumia root medicinal extract solution that 20ml is settled to after sufficiently milling uniformly to get concentration.It will be prepared
The Artemether solution and concentration that concentration is 20mg/ml are that 1:1 is mixed by volume for 0.75g crude drug/ml colguhoumia root medicinal extract solution
To get the solution of required proportion (A1:B2), (the final concentration of 10mg/ml of Artemether, the final concentration of 0.375g of colguhoumia root are raw after conjunction
Medicine/ml).
Being settled to 20ml after taking 400mg Artemether, addition 0.5%CMC-Na solution sufficiently to mill uniformly to get concentration is
The Artemether solution of 20mg/ml;2.2g colguhoumia root medicinal extract (content is 13.64g crude drug/g medicinal extract) is taken, 0.5%CMC- is added
Na solution is settled to 20ml to get concentration after sufficiently milling uniformly be 1.5g crude drug/ml colguhoumia root medicinal extract solution.It will prepare
The Artemether solution and concentration that good concentration is 20mg/ml are 1.5g crude drug/ml colguhoumia root medicinal extract solution 1:1 by volume
To get solution (the final concentration of 10mg/ml of Artemether, the final concentration of 0.75g of colguhoumia root of required proportion (A1:B1) after mixing
Crude drug/ml).
200mg Artemether is taken, is added after 0.5%CMC-Na solution is sufficiently milled uniformly and is settled to 20ml to get required
Artemether solution (A1) (the final concentration of 10mg/ml of Artemether).
1.1g colguhoumia root medicinal extract (content is 13.64g crude drug/g medicinal extract) is taken, 0.5%CMC-Na solution is added and sufficiently grinds
Mill uniformly after be settled to 20ml to get required colguhoumia root medicinal extract solution (B1) (the final concentration of 0.75g crude drug of colguhoumia root/
ml)。
2.2.2 experimental method
After quarantine and adaptive feeding, choose 56 qualified mouse of health, weight be no more than average weight ±
20%, it is randomly divided into 7 groups, i.e. blank control group, Artemether and tripterygium hypoglaucum hutch A2:B2 group, Artemether and tripterygium hypoglaucum hutch A1:B2 group, wormwood artemisia
Methyl ether and tripterygium hypoglaucum hutch A2:B1 group, Artemether and tripterygium hypoglaucum hutch A1:B1 group, Artemether A1 group, tripterygium hypoglaucum hutch B1 group, every group of 8 (male and female
It is fifty-fifty).After grouping, corresponding drug is given in stomach-filling to each medicine group respectively, and blank control group gives the purified water of respective volume,
Administered volume is 0.2ml/10g weight, once a day, successive administration 7 times.
It is administered the 3rd day, each group mouse uniformly smears 0.5% 2,4-dinitrofluorobenzene acetone olive oil at abdomen unhairing
Solution (25ul/ is only) sensitization, is administered the 4th day and strengthens 1 time.
After last dose 1 hour, 0.2%2,4- dinitrofluorobenzene acetone olive oil solution (20ul/ is only) is uniformly applied to
Mouse right ear, left ear smear matrix (acetone olive oil), and model control group smears matrix.After 30 minutes, cervical dislocation puts to death mouse,
Left and right auricle is cut, auricle is removed with punch (diameter 8mm), claims left and right ear weight, calculate swelling and swelling inhibiting rate, calculate
Formula is as follows:
Swelling=auris dextra weight-Zuo Erchong
Swelling inhibiting rate=(model group swelling-medicine group swelling)/model group swelling × 100%
2.2.3 experimental result
Such as table 2, the different ratio of Artemether and tripterygium hypoglaucum hutch has apparent inhibition to ear swelling caused by 2,4-dinitrofluorobenzene
Effect, prompts it to have apparent inhibiting effect to delayed hypersensitive reaction caused by 2,4-dinitrofluorobenzene, and proportion effect is better than list
With.
Table 2
Group |
Ratio |
Swelling |
Swelling inhibiting rate |
Blank control |
/ |
0.0070±0.0020 |
/ |
Artemether: tripterygium hypoglaucum hutch |
A2:B2 |
0.0057±0.0036 |
18.57 |
Artemether: tripterygium hypoglaucum hutch |
A1:B2 |
0.0067±0.0022 |
4.29 |
Artemether: tripterygium hypoglaucum hutch |
A2:B1 |
0.0049±0.0019 |
30.00 |
Artemether: tripterygium hypoglaucum hutch |
A1:B1 |
0.0061±0.0023 |
12.86 |
Artemether |
A1 |
0.0067±0.0025 |
4.29 |
Tripterygium hypoglaucum hutch |
B1 |
0.0068±0.0026 |
2.86 |
The influence of 2.3 pairs of experimental animal model of CFA induced adjuvant arthritis in rats
2.3.1 drug is prepared
Test group medication: taking 200mg Artemether, is added after 0.5%CMC-Na solution is sufficiently milled uniformly and is settled to 20ml,
The Artemether solution for being 10mg/ml up to concentration;1.1g colguhoumia root medicinal extract (content is 13.64g crude drug/g medicinal extract) is taken, is added
0.5%CMC-Na solution is settled to 20ml to get concentration after sufficiently milling uniformly be 0.75g crude drug/ml colguhoumia root medicinal extract
Solution.By prepared concentration be 10mg/ml Artemether solution and concentration be that 0.75g crude drug/ml colguhoumia root medicinal extract is molten
To get the solution of required proportion (A4:B4), (the final concentration of 5mg/ml of Artemether, colguhoumia root are whole after liquid 1:1 mixing by volume
Concentration is 0.375g crude drug/ml).
Being settled to 20ml after taking 200mg Artemether, addition 0.5%CMC-Na solution sufficiently to mill uniformly to get concentration is
The Artemether solution of 10mg/ml;2.2g colguhoumia root medicinal extract (content is 13.64g crude drug/g medicinal extract) is taken, 0.5%CMC- is added
Na solution is settled to 20ml to get concentration after sufficiently milling uniformly be 1.5g crude drug/ml colguhoumia root medicinal extract solution.It will prepare
The Artemether solution and concentration that good concentration is 10mg/ml are 1.5g crude drug/ml colguhoumia root medicinal extract solution 1:1 by volume
To get the solution of required proportion (A4:B3), (the final concentration of 5mg/ml of Artemether, the final concentration of 0.75g of colguhoumia root are raw after mixing
Medicine/ml).
Being settled to 20ml after taking 400mg Artemether, addition 0.5%CMC-Na solution sufficiently to mill uniformly to get concentration is
The Artemether solution of 20mg/ml;It takes 1.1g colguhoumia root medicinal extract (13.64g crude drug/g medicinal extract), 0.5%CMC-Na solution is added
It is 0.75g crude drug/ml colguhoumia root medicinal extract solution that 20ml is settled to after sufficiently milling uniformly to get concentration.It will be prepared
The Artemether solution and concentration that concentration is 20mg/ml are that 1:1 is mixed by volume for 0.75g crude drug/ml colguhoumia root medicinal extract solution
To get the solution of required proportion (A3:B4), (the final concentration of 10mg/ml of Artemether, the final concentration of 0.375g of colguhoumia root are raw after conjunction
Medicine/ml).
Being settled to 20ml after taking 400mg Artemether, addition 0.5%CMC-Na solution sufficiently to mill uniformly to get concentration is
The Artemether solution of 20mg/ml;2.2g colguhoumia root medicinal extract (content is 13.64g crude drug/g medicinal extract) is taken, 0.5%CMC- is added
Na solution is settled to 20ml to get concentration after sufficiently milling uniformly be 1.5g crude drug/ml colguhoumia root medicinal extract solution.It will prepare
The Artemether solution and concentration that good concentration is 20mg/ml are 1.5g crude drug/ml colguhoumia root medicinal extract solution 1:1 by volume
To get solution (the final concentration of 10mg/ml of Artemether, the final concentration of 0.75g of colguhoumia root of required proportion (A3:B3) after mixing
Crude drug/ml).
200mg Artemether is taken, is added after 0.5%CMC-Na solution is sufficiently milled uniformly and is settled to 20ml to get required
Artemether solution (A3) (the final concentration of 10mg/ml of Artemether).
1.1g colguhoumia root medicinal extract (content is 13.64g crude drug/g medicinal extract) is taken, 0.5%CMC-Na solution is added and sufficiently grinds
Mill uniformly after be settled to 20ml to get required colguhoumia root medicinal extract solution (B3) (the final concentration of 0.75g crude drug of colguhoumia root/
ml)。
2.3.2 experimental method
Reference literature method carries out model preparation: after quarantine and adaptive feeding, the qualified rat of quarantine (being stayed 8
Rat is not processed, half male and half female) Yu Youhou toes intracutaneous injection CFA 0.1mL/ is only.
The 14th day after injection, the successful rat of modeling type is selected to be randomly divided into 7 groups, i.e. model comparison according to Articular swelling
Group, Artemether and tripterygium hypoglaucum hutch A4:B4 group, Artemether and tripterygium hypoglaucum hutch A3:B4 group, Artemether and tripterygium hypoglaucum hutch A4:B3 group, Artemether with
Tripterygium hypoglaucum hutch A3:B3 group, Artemether A3 group, tripterygium hypoglaucum hutch B3 group, every group 8 (half male and half female);It is blank that another 8, which do not process animal,
Control group.After grouping, corresponding drug is given in stomach-filling to each medicine group respectively, and blank control group, model control group are given accordingly
The purified water of volume, administered volume are 1ml/100g weight, by daily single, successive administration 21 days.
It (the 0th day) and is administered before modeling, before administration and measures the left back foot (secondary disease of all rats respectively on the 7th, 14,21 day
Become side) and ankle-joint diameter 1 time of right metapedes, calculate Articular swelling.Calculation formula is as follows:
Rat ankle joint diameter before rat ankle joint diameter-modeling after Articular swelling=administration
2.3.3 experimental result
Such as table 3, the different ratio of Artemether and tripterygium hypoglaucum hutch has certain therapeutic effect to rat assist agent arthritis, prompts
It has apparent therapeutic effect to rat assist agent arthritis subsequent articular swelling, and proportion effect, which is better than, to be applied alone.
Table 3
The prompt of Pharmacodynamics screening result of study with rat arthritis model for main evaluation criterion, Artemether and tripterygium hypoglaucum hutch
A4:B3 proportion (dosage be equal to mouse A2:B1 proportion) play the role of it is obvious, should with compare DNFB cause mouse it is tardy
Hypersensitivity and mouse hemolytic antibody, which generate experiment, apparent inhibiting effect.
3, acute toxicity testing result
Experimental method: after quarantine and adaptive feeding, 60 qualified mouse of health are chosen, weight is no more than average
± the 20% of weight is randomly divided into 3 groups, every group 20 (half male and half female).By group, relative medicine is given in stomach-filling respectively, gives medicine body
Product is 0.4ml/10g weight, is administered once.
Tripterygium hypoglaucum hutch 200g crude drug/kg: it dead 18, survives 2;
Artemether 4g/kg: dead 19, it survives 1;
Tripterygium hypoglaucum hutch 200g crude drug/kg+ Artemether 4g/kg: it dead 17, survives 3.
It can be seen that the faint decline of toxicity after tripterygium hypoglaucum hutch and Artemether combination, from result above it can also be seen that, fire when administration
It should not be more than the above-mentioned amount having compared with high lethality rate colored and Artemether dosage.