The present invention is further elaborated with the biological activity test example by the following examples:
Embodiment 1
Pharmaceutical composition of the present invention is made by following proportion raw material medical material:
Radix Glycyrrhizae 700g Radix Platycodonis 250g Rhizoma Belamcandae 280g
Fructus Arctii 200g Herba Menthae 200g Periostracum Cicadae 320g
Radix Scrophulariae 500g Radix Paeoniae Rubra 600g Herba Schizonepetae 330g
Preparation method:
Radix Glycyrrhizae, Radix Platycodonis, Fructus Arctii, Rhizoma Belamcandae, Radix Scrophulariae, Radix Paeoniae Rubra, Periostracum Cicadae are pulverized, decoct with water twice, each 90 minutes, decocting liquid merged.Herba Schizonepetae, Herba Menthae are used steam distillation, isolate volatile oil, and remaining distillate and decocting liquid merge filtration.Filtrate decompression is concentrated into relative density the be 1.15 extractum liquid of (80 ℃ of heat are surveyed), puts and is chilled to room temperature, adds ethanol and is adjusted to and contains alcohol amount 60% (w/w), and the limit edged stirs, and leaves standstill 24 hours.Get supernatant, being evaporated to relative density is the thick extractum of 1.36 (80 ℃ of heat are surveyed).With separate Herba Schizonepetae-Herba Menthae Haplocalycis volatile oil be added dropwise to saturated beta-schardinger dextrin-aqueous solution while stirring, add the back and continue to stir 2 hours, places 16 hours after-filtration, drying, beta-cyclo dextrin included compound.Thick extractum is mixed with beta-cyclo dextrin included compound, get pharmaceutical composition 720g of the present invention.
Pharmaceutical composition 720g
Icing Sugar 450g
Dextrin 200g
According to the conventional preparation technology of granule, Icing Sugar and dextrin are added pharmaceutical composition, mix homogeneously is made granule, 60 ℃ of dryings, packing is promptly.
The biological activity test example
Example 1: bacteriostatic test in the body
Animal: 100 of Kunming mouses
Medicine-feeding way: irritate stomach
The administration group: little, in, heavy dose of (being equivalent to 0.6g, 1.7g, 5.0g extract powder respectively)
Matched group: SHUANGHUANGLIAN KOUFUYE 25ml/Kg
Test method: 10 every group of mices, administration every day 1 time continuous 5 days, is infected staphylococcus aureus, the death toll of observing mice in 24 hours in the 30min mouse peritoneum after the last administration.The results are shown in following table:
Group | Dosage (g/kg) | Number of animals (only) | Death toll (only) | Mortality rate (%) |
The heavy dose of group of dosage group SHUANGHUANLIAN group in the normal saline group small dose group | - 0.6 1.7 5.0 25ml/Kg | 20 20 20 20 20 | 18 19 14 9 11 | 90 95 70 45
** 55
** |
Compare with the normal saline group,
*P<0.05.
The result shows that the heavy dose of group of pharmaceutical composition of the present invention can reduce mortality rate to the infection of staphylococcus aureus mice, has the certain protection effect.
Example 2: antiinflammatory test
(1) to the swollen outgrowth influence of rat granuloma
Animal: 50 of male SD rats
Medicine-feeding way: irritate stomach
The administration group: little, in, heavy dose of (being equivalent to 0.4g, 1.3g, 3.8g extract powder respectively)
Matched group: aspirin 0.1g/Kg
Test method: 10 every group of rats, cotton balls is implanted in the anesthesia back, and postoperative is gastric infusion respectively, administration every day 1 time, continuous 7 days.Put to death rat on the 8th day, calculate the granuloma net weight, the results are shown in following table:
Group | Dosage (g/Kg) | Number of animals (only) | Granuloma dry weight (mg/100g) | Suppression ratio (%) |
The heavy dose of group of dosage group aspirin group in the normal saline group small dose group | - 0.4 1.3 3.8 0.1 | 9 10 9 10 9 | 49.1±13.3 36.8±10.3
* 33.4±13.2
* 33.0±10.9
* 30.7±9.4
** | 25.05 31.98 32.79 37.47 |
Compare with matched group,
*P<0.05,
*P<0.01.
Annotate: 3 rats are destroyed because of operation suture thread, and the cotton balls that causes implanting is lost, and can't calculate, and therefore exclude result of the test.
The result shows, compare with the normal saline matched group, each dosage group of pharmaceutical composition of the present invention all can significantly suppress by the rat chronic granulation tissue hyperplasia due to the implantation cotton balls, and suppression ratio is respectively 25.05%, 31.98%, 32.79%, illustrates that it has certain inhibitory action to chronic inflammatory disease.
(2) influence of xylol induced mice auricle inflammation
Animal: 50 of Kunming mouses
Medicine-feeding way: irritate stomach
The administration group: little, in, heavy dose of (being equivalent to 0.6g, 1.7g, 5.0g extract powder respectively)
Matched group: aspirin 0.1g/Kg
Test method: 10 every group of mices, administration every day 1 time continuous 3 days, after the last administration 1 hour, is smeared to mouse ear with dimethylbenzene.Put to death mice after 1 hour, calculate the auricle edema value, the results are shown in following table:
Group | Dosage (g/Kg) | Auricle edema value (g) | Auricle edema rate (%) | Suppression ratio (%) |
The heavy dose of group of dosage group aspirin group in the normal saline group small dose group | - 0.6 1.7 5.0 0.1 | 10.6±2.2 8.4±2.5 8.2±2.1 7.9±2.0 7.3±2.1 | 166.4±43.8 121.8±37.0
* 124.4±43.5
* 117.5±38.9
* 109.9±37.6
** | 26.8 25.2 29.4 34.0 |
Compare with matched group,
*P<0.05,
*P<7.30.01.
The result shows, compares with the normal saline matched group, and each dosage group of pharmaceutical composition of the present invention all can significantly suppress the Mice Auricle inflammation due to the dimethylbenzene, and suppression ratio is respectively 26.8%, 25.2%, 29.4%.
(3) on Carrageenan causes the influence of rat paw edema
Animal: 50 of male SD rats
Medicine-feeding way: irritate stomach
The administration group: little, in, heavy dose of (being equivalent to 0.4g, 1.3g, 3.8g extract powder respectively)
Matched group: aspirin 0.1g/Kg
Test method: 10 every group of mices, gastric infusion respectively, after the administration 1 hour, in the right back sufficient sole of the foot injection carrageenin of rat, cause scorching back 1,2,4,6 hour, measure sufficient sole of the foot volume respectively, calculate swelling value, swelling rate, the results are shown in following table:
Group | Dosage (g/Kg) | Swelling value (ml) |
1h | 2h | 4h | 6h |
The heavy dose of group of dosage group aspirin group in the normal saline group small dose group | - 0.4 1.3 3.8 0.1 | 0.41±0.15 0.35±0.13 0.22±0.13
** 0.24±0.13
* 0.25±0.16
| 0.73±0.13 0.50±0.9
** 0.49±0.16
** 0.47±0.16
** 0.41±0.15
** | 0.55±0.15 0.42±0.12 0.34±0.15
** 0.33±0.16
** 0.28±0.17
** | 0.37±0.15 0.31±0.12 0.26±0.15 0.30±0.15 0.24±0.13 |
Compare with matched group,
*P<0.05,
*P<0.01.
Group | Dosage (g/Kg) | Swelling rate (%) |
1h | 2h | 4h | 6h |
The heavy dose of group of dosage group aspirin group in the normal saline group small dose group | - 2.5 5 10 0.1 | 44.0±20.1 37.2±16.9 26.0±17.6
* 26.5±16.2
* 25.5±16.7
* | 76.4±19.8 53.5±25.3
* 55.6±23.3
* 52.2±22.7
* 42.9±18.3
* | 58.2±21.8 44.7±18.2 40.1±21.5 37.3±21.6
* 30.6±20.9
* | 39.7±19.8 33.7±17.7 29.8±19.3 34.3±20.3 26.3±19.7 |
Compare with matched group,
*P<0.05,
*P<0.01.
The result shows, compares with the normal saline group, and each dosage group of pharmaceutical composition of the present invention all can obviously suppress the rat paw edema due to the carrageenin, onset in 1 hour after the general administration, and effect lasted till 4 hours.
(4) to the influence of mouse peritoneal capillary permeability
Animal: 50 of Kunming mouses
Medicine-feeding way: irritate stomach
The administration group: little, in, heavy dose of (being equivalent to 0.6g, 1.7g, 5.0g extract powder respectively)
Matched group: aspirin 0.1g/Kg
Test method: 10 every group of mices, administration every day 1 time, continuous 3 days, after the last administration 1 hour, behind the mouse tail vein injection azovan blue normal saline, lumbar injection acetic acid normal saline, put to death mice after 30 minutes, measure the optical density value of abdominal cavity cleaning mixture, the results are shown in following table:
Group | Dosage (g/Kg) | Optical density value (OD) |
The heavy dose of group of dosage group aspirin group in the normal saline group small dose group | - 0.6 1.7 5.0 0.1 | 0.370±0.137 0.264±0.162 0.190±0.106
** 0.196±0.112
** 0.211±0.111
* |
Compare with the normal saline group,
*P<0.05,
*P<0.01.
The result shows, with the normal saline group relatively, the mouse peritoneal capillary permeability due to the big or middle dosage group of the pharmaceutical composition of the present invention Dichlorodiphenyl Acetate increases obvious inhibitory action.
Example 3: analgesic test
The influence of Dichlorodiphenyl Acetate induced mice writhing response
Animal: 50 of Kunming mouses
Medicine-feeding way: irritate stomach
The administration group: little, in, heavy dose of (being equivalent to 0.6g, 1.7g, 5.0g extract powder respectively)
Matched group: aspirin 0.2g/Kg
Test method: 10 every group of mices, after the administration 1 hour, mouse peritoneal injection acetum, that observes each treated animal in 15 minutes turns round the body number of times, the results are shown in following table:
Group | Dosage (g/Kg) | Turn round the body number of times | Suppression ratio (%) |
The heavy dose of group of dosage group aspirin group in the normal saline group small dose group | - 0.6 1.7 5.0 0.2 | 47.5±12.4 36.6±10.1
* 34.0±9.3
* 32.6±10.5
** 31.5±12.2
** | 22.9 28.4 31.4 33.7 |
Compare with matched group,
*P<0.05,
*P<0.01.
The result shows, with the normal saline matched group relatively, the mouse writhing reaction due to each dosage group Dichlorodiphenyl Acetate of pharmaceutical composition of the present invention has obvious inhibitory action, suppression ratio is respectively 22.9%, 28.4%, 31.4%, illustrates that it has certain analgesic activity.
Example 4: expectorant test
Influence to the phenol red discharge rate of mice trachea
Animal: 50 of Kunming mouses
Medicine-feeding way: irritate stomach
The administration group: little, in, heavy dose of (being equivalent to 0.6g, 1.7g, 5.0g extract powder respectively)
Matched group: ammonium chloride 1.0g/Kg
Test method: 10 every group of mices, after the administration 1 hour, mouse peritoneal injection phenol red solution was put to death mice after 30 minutes, measured the optical density value of trachea cleaning mixture, the results are shown in following table:
Group | Dosage (g/Kg) | Phenol red output (μ g/ml) | Rate of increase (%) |
The heavy dose of group of dosage group sodium chloride group in the normal saline group small dose group | - 0.6 1.7 5.0 1.0 | 0.48±0.29 0.78±0.32
* 0.81±0.37
* 0.88±0.32
** 1.04±0.27
** | 62.5 68.8 83.3 116.7 |
Compare with the normal saline group,
*P<0.05,
*P<0.01.
The result shows, compares with the normal saline matched group, and each dosage group of pharmaceutical composition of the present invention all can obviously promote the phenol red discharge of mice trachea, illustrates that it has certain phlegm-dispelling functions.
From the biological activity test result as can be seen, that pharmaceutical composition of the present invention has is antibiotic significantly, antiinflammatory, phlegm-dispelling functions, thus acute pharyngitis is had significant therapeutic effect, and can prevent or reduce acute pharyngitis protracted course of disease or outbreak repeatedly, and have no adverse reaction.