CN1526423A - Medicine composition for treating gastric ulcer, stomachache and superficial gastritis and its prepn and use - Google Patents
Medicine composition for treating gastric ulcer, stomachache and superficial gastritis and its prepn and use Download PDFInfo
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Abstract
The present invention is one kind of Chinese medicine composition for treating gastric ulcer, stomachache and superficial gastritis and its preparation process and use. The Chinese medicine composition is prepared with chinaberry fruit, vinegar processed corydalis tuber, redflower anise leaf and aucklandia root. It has high pain-relieving effect, quick acting, wide pain-relieving range, and no medicine dependence and resistance.
Description
Technical field
The present invention relates to a kind of treatment gastric ulcer, stomachache or superficial gastritis pharmaceutical composition and its production and use.
Background technology
The stomach pain symptom comes across in the numerous disease, brings very big misery to the patient, influences work, study and daily life even.Common Western medicine analgesic has dependency, toleration, and has other side effect, and the application aims to provide the medicine that a kind of analgesic effect better, also can be treated gastric ulcer, superficial gastritis.
Summary of the invention
The application's compositions is made by following materials of weight proportions: Fructus Toosendan 3-5, Rhizoma Corydalis (vinegar system) 3-5, the anistree leaf 1-2 of Flos Carthami, Radix Aucklandiae 1-2, preferred weight proportion is: Fructus Toosendan 3, Rhizoma Corydalis (vinegar system) 3, the anistree leaf 2 of Flos Carthami, the Radix Aucklandiae 2, but the compositions of not exclusively making by the proportioning in the above scope also is effective.The preferred Saussurea lappa Clarke of the Radix Aucklandiae wherein.
The application's compositions can adopt the conventional method of galenic pharmacy to make regular dosage form, and as powder, tablet, electuary, oral liquid etc., preferred dosage form is a tablet.
The preparation method of tablet or electuary can for: get raw material in proportion: the anistree leaf of Fructus Toosendan, Rhizoma Corydalis (vinegar system), Flos Carthami, Saussurea lappa Clarke.Fructus Toosendan, Saussurea lappa Clarke are added suitable quantity of water decoct, gained filtrate merges, and it is standby to be concentrated into extractum; Rhizoma Corydalis (vinegar system), the anistree leaf of Flos Carthami add an amount of 40-95% ethanol liquid reflux, extract,, and gained filtrate merges, decompression recycling ethanol, and it is standby to be concentrated into extractum.Two step extractum merge, and add conventional excipients, and dry method or wet granulation are made electuary, or further tabletting is made tablet.The used quantity of solvent of each extraction medical material is 8-10 a times of medical material weight.Medical material preferably extracts 2 times.Extract the used ethanol of medical material and preferably use 50% ethanol.
The method of quality control of compositions
Qualitative identification method: identify the Radix Aucklandiae, Quercetin, tetrahydropalmatine in the compositions
(1) it is an amount of to get compositions, adds that petroleum ether (60-90 ℃) is an amount of to be extracted, and divides and gets the ether layer, is concentrated into small size, as need testing solution; It is an amount of that other gets Radix Aucklandiae control medicinal material powder, adds Petroleum ether extraction, divides and get the ether layer, is concentrated into small size medical material solution in contrast.Test according to thin layer chromatography (35 pages of 1995 editions appendix of Chinese Pharmacopoeia), each is an amount of to draw above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (60-90 ℃)-ethyl acetate one benzene (14: 3: 3) is developing solvent, launch, taking-up is dried, and spray is with 5% vanillin dilute sulfuric acid solution.Heat after several minutes, in the test sample chromatograph with the corresponding position of control medicinal material chromatograph on, show identical blue spot.
(2) it is an amount of to get compositions, add ethanol extraction, get filtrate, fling to ethanol or reclaim ethanol, with suitable quantity of water with residue suspendible or dissolving, with ethyl acetate extraction, collect acetic acid ethyl acetate extract, remove and desolvate, residue adds 10% hydrochloric acid 10ml in water-bath hydrolysis 1.5-3 hour, preferred 2 hours, filter the filtrate evaporate to dryness; Residue adds dissolve with ethanol, as need testing solution; Other gets the Quercetin reference substance, adds alcoholic solution product solution in contrast.According to thin layer chromatography (35 pages of 1995 editions appendix of Chinese Pharmacopoeia) test, each is an amount of to draw need testing solution and reference substance solution, put respectively on same silica gel g thin-layer plate, and be that developing solvent launches with toluene-Ethyl formate-formic acid (5: 4: 1), dry.Natural light is observed down, in the test sample chromatograph with the corresponding position of reference substance chromatograph on, show identical yellow spotting.
(3) it is an amount of to get compositions, adds a small amount of distilled water and equivalent strong aqua ammonia moistening medicated powder, adds an amount of chloroform extraction again, collects extracting solution, is concentrated into small size, makes test liquid; Other gets the tetrahydropalmatine reference substance, and it is an amount of to add ethanol, makes reference substance solution; Test according to thin layer chromatography (35 pages of 1995 editions appendix of Chinese Pharmacopoeia), each is an amount of to draw above-mentioned two kinds of solution, putting respectively on same silica gel g thin-layer plate, is developing solvent with petroleum ether-chloroform-methanol (10: 6: 1), launches, takes out, dries, sprays with bismuth potassium iodide test solution.In the test sample chromatograph with the corresponding position of reference substance chromatograph on, show the speckle of same color.
The above extraction comprises backflow and ultrasonic extraction.
Assay: the tetrahydropalmatine in the titration measuring compositions
It is an amount of to get compositions, and accurate the title decides, and porphyrize is put in the conical flask, add the ammonia appropriate basesization, shake up, it is an amount of to add benzene, hour filters surplus placing 10, and it is an amount of that residue adds benzene again, shake up, surplus placing 10 hour, filter merging filtrate, filtrate is moved in the separatory funnel, with the sulphuric acid extraction of 0.1-0.5mol/L three times, merge extractive liquid, adds ammonia and transfers to alkalescence, with chloroform extraction for several times, combined chloroform, with the rearmounted water bath method of distilled water wash, residue adds the chloroform dissolving, evaporate to dryness, the dissolving of reuse minimum of chloroform, the accurate sulphuric acid 25ml that adds 0.01mol/L puts water-bath and flings to chloroform, put and be chilled to room temperature, 3 of the red indicators of methylate, with the caustic lye of soda titration of 0.02mol/L, the result proofreaies and correct with blank assay.
Every 1ml 0.01mol/L sulphuric acid is equivalent to 7.1084mg Rhizoma Corydalis second rope.
Pharmacodynamic study proves: the prepared tablet golden red tablet for alleviating pain of the application's compositions removes has the gastric emptying of delaying; Reduce gastric secretion and total acid output, suppress outside the effect of pepsinia, and have analgesia, spasmolytic, antiinflammatory, antiulcer action.This result shows that the golden red tablet for alleviating pain can regulate gastrointestinal function, plays the effect of depressed liver-energy dispersing and QI regulating and stomach and alleviating pain, can treat gastric ulcer, stomachache or superficial gastritis.
Pharmacodynamic test of active extract
Experiment material
1, medicine: golden red tablet for alleviating pain: form (seeing data three) by Fructus Toosendan, Rhizoma Corydalis (vinegar system), the anistree leaf of Flos Carthami, Saussurea lappa Clarke, every contains crude drug 2.15g, with tablet (plain sheet) pulverization, be desired concn (PH5.5) with distilled water diluting during test, the time spent shakes up.Rotundine: 40mg/ sheet, Kweiyang Golden Bridge pharmaceutical factory produces, prepare the same, lot number 881014, meperidine hydrochloride inj (pethidine): 50mg/ml, Yichang pharmaceutical factory, lot number 881024,36% acetic acid, sodium hydroxide (AR) Chengdu chemical reagent factory, lot number 8809160 carrageenin (carrageenan) and serotonin (5-HT), sigman product.The little Chaohu of Rhizoma Corydalis tablet for alleviating pain (be called for short the Rhizoma Corydalis sheet, form) peace pharmaceutical factory by Rhizoma Corydalis, the Radix Angelicae Dahuricae, lot number 880816, histamine phosphate and acetyl chloride gallbladder treasure, Shanghai Inst. of Biochemistry, Chinese Academy of Sciences 8510212.
2, instrument: 721 type spectrophotometers: Shanghai the 3rd analytical tool factory product.L7-IIIA type automatic constant-temperature isolated organ tester: Chuanbei Medical College makes.
3, animal: Kunming kind healthy mice, body weight 18-22g, the wistar healthy white rat, body weight 180-220g, big mice male and female dual-purpose, animal housing provides by Guiyang Medical College.
Method and result
(1) the golden red tablet for alleviating pain is to the influence (colorimetry) of mice gastric emptying motion
Get 30 of body weight 20-22g healthy mices, (as follows) is divided into three groups at random, every group 10, male and female half and half, fasting 12h (but freely drinking water), for the interference of the color contrast color of avoiding Chinese medicine gives each treated animal after all will Chinese medicine filtering, drug dose is 0.1ml/10g (drug dose of following mice all herewith).40min after the administration, every Mus ig0.2ml0.1% methyl orange liquid.20min post-tensioning neck is put to death animal, cut open the belly, win stomach, place the small beaker that contains the 10ml distilled water, cut off along greater gastric curvature, gastric content is fully washed in distilled water with little shears, transfer pH value to 6.0-6.5 with NaHCO3 liquid, pour graduated centrifuge tube into, with 2000rpm, centrifugal 10min gets supernatant with 721 type spectrophotometer colorimetrics (going into 420nm).The OD value is surveyed in the distilled water zeroing.Add in the 10ml distillation with 0.1% methyl orange 0.2ml and to shake up, survey OD value,, be calculated as follows methyl orange stomach residual rate, and (following test all with) checked in " t " that takes statistics as radix methyl orange optical density.
Result's (table 1) shows golden red tablet for alleviating pain 8g/kg and 10g/kg group, and relatively there were significant differences with matched group, can improve the methyl orange residual rate, the tool mice gastric emptying that slows down.
Table 1 golden red tablet for alleviating pain is to the influence of mice gastric emptying (M ± SD)
Group number of animals (n) dosage (g/kgd) methyl orange residual rate (%)
NS 10 equal-volumes 24.6 ± 4.3
Golden red tablet for alleviating pain 10 8.0 31.1 ± 6.7*
Golden red tablet for alleviating pain 10 10.0 38.3 ± 7.5**
Golden red tablet for alleviating pain and NS group be * P＜0.05 * * P＜0.01 * * * P＜0.001 relatively, and the NS group is for giving the matched group (as follows) of distilled water.
(2) the golden red tablet for alleviating pain is to the influence of rat gastric acid, pepsinia
Get 24 of body weight 180 ± 22g Mus, be divided into three groups at random, male and female half and half, single cage is fed, and fasting 24h freely drinks water, behind the slight numb sour rat of ether, the abdominal part cropping, routine disinfection skin, median line is made the 2.5cm otch in the xiphoid-process lower edge.Carefully mention stomach, avoid blood vessel suture ligation at pylorus and duodenum bound fraction.Inject golden red tablet for alleviating pain liquid 8g/kg and 10g/kg (1ml/100g) by duodenum immediately, matched group is given equal-volume water with method, puts back to behind the sew up wound in the cage, and water is prohibited in the postoperative fasting.Take off cervical vertebra behind the 5h and put to death, get stomach, collection gastric juice, is drawn supernatant and is calculated the gastric juice amount with the centrifugal 15min of 3000rpm in graduated centrifuge tube, parallel gastric analysis.
1. gastric acidity determination: get clear gastric juice 2ml and put and add start going to a nursery each 2 of Fo Shi and phenolphthalein indicators in the small beaker.Be cherry-red when containing free acid.With the burette titration that fills 0.02mol/L NaoH, and constantly shake triangular flask, occur till the ginger color, be the free acid titration end-point to red disappearance.Record consumes the ml number of NaoH liquid, till continuation is not taken off to blush with the NoaH drop is fixed again, be the total acidity titration end-point, remember total ml number of the 0.02mol/L NaoH that is consumed for twice, the result calculates: ml number * 10 of the NoaH liquid that free acid (mEq/L)=free acid titration end-point consumes.Total acidity (NaoH liquid ml number * 10 that twice titration of mEq/L=consumes.Total acid output (meq/h)=total acidity * gastric juice amount hourly, the result shows (table 2) golden red tablet for alleviating pain 10g/kg group to gastric secretion, and total acid output has the reduction effect, and the 5g/kg group does not then have obviously influence.
2. pepsin is measured (wheat Te Shi capillary glass-tube method): the capillary glass-tube of internal diameter 0.5mm even thickness is cut into 10cm length, clean, dry, the Ovum Gallus domesticus album of getting one piece of egg fully stirs evenly the back filtered through gauze, utilize siphonage capillaceous, fill Ovum Gallus domesticus album (the pipe planted agent does not have bubble) in the pipe, be placed in 85 ℃ of water and make protein coagulation, the storage refrigerator is standby.
(bottle adds 0.05mol/L HCL liquid 15ml to get the triangular flask that rat gastric juice 1ml puts into 50ml during test, shake up), put into two of above-mentioned protein pipe again, filled in bottleneck, hatch 24h in 37 ℃ of calorstats, take out the length (mm) of back, ask its meansigma methods with the value of four ends with the transparent part in chi measurement protein pipe two ends.Pepsin activity unit=meansigma methods 2 * 16, the pepsin output,, (X of active unit/h=pepsin activity unit gastric juice amount hourly, result showed that (table 3) golden red tablet for alleviating pain 10g/kg has inhibitory action to pepsinia.
Table 2 golden red tablet for alleviating pain is to the influence of rat gastric juice, gastric acid secretion (M ± SD)
The total acid output of group dosage (g/kgd) gastric juice amount (mEq/L) free acid (mEq/L) total acidity (mEq/L)
(μmEq/L)
NS equal-volume (8) 9.18 ± 2.41 98.6 ± 11.4 127.6 ± 14.7 231.4 ± 56.4
Golden red tablet for alleviating pain 5.0 (8) 8.69 ± 2.13 102.0 ± 6.2 130.8 ± 7.1 218.9 ± 62.7
Golden red tablet for alleviating pain 10.0 (8) 6.21 ± 2.32* 104.6 ± 11.3 125.8 ± 12.7 173.9 ± 53.9**
In the Δ bracket is the animal number of elements
Table 3 golden red tablet for alleviating pain is to the influence of pepsin activity and output thereof (M ± SD)
Group dosage (g/kgd) pepsin activity pepsin output
(active unit/ml)) (active unit/ml)
NS equal-volume (8) 223.0 ± 37.0 438.9 ± 79.8
Golden red tablet for alleviating pain 5.0 (8) 237.0 ± 51.6 415.8 ± 96.8
Golden red tablet for alleviating pain 10.0 (8) 239.8 ± 63.9 310.2 ± 89.7**
(3) the golden red tablet for alleviating pain is to the influence of mice intestinal propelling rate: taking body weight is 30 of 21.08 ± 0.74g mices, male and female half and half, be divided into three groups, each organizes administration every day (ig) once, successive administration 3d, give in the medicine by 1: 1 adding 20% active carbon in each group of 4d behind the fasting 22h before testing, each medicine capacity of ig is 0.2ml/10g, and 30min puts to death behind the ig, cuts open the belly, the careful separation mesentery, the clip pylorus is to the intestinal tube of ileocecus
(4)Be put in the big square plate, gently that intestinal tube is stretching, survey intestinal segment total length and distance (this distance as prepared Chinese ink in the propulsive distance of enteral), with following formula calculating intestinal propelling rate (%) from pylorus to the prepared Chinese ink forward position.The result shows that (table 4) golden red tablet for alleviating pain can significantly delay the intestinal propulsion rate.
Table 4 golden red tablet for alleviating pain is to the influence of mice intestinal propelling rate (M ± SD)
Group dosage (g/kgd) number of animals (only) intestinal propelling rate (%)
NS - 10 79.36±11.56
Golden red tablet for alleviating pain 5.0 10 62.18 ± 10.98*
Golden red tablet for alleviating pain 10.0 10 51.21 * 11.23**
(4) analgesic test: 1, to the influence of acetic acid induced mice writhing response: press document
(5), revise a little, 50 of mices, body weight is 19.8 ± 1.37 (SD) male and female half and half, is divided into five groups, each organize behind the ig 1h (im pethidine min) immediately ip0.6 acetic acid 0.2ml/ only, the number of times of writhing response in the acetic acid 15min given in record.Result's (table 5) shows that golden red tablet for alleviating pain two dosage groups all have the effect of remarkable inhibition writhing response, and relatively there were significant differences and golden red tablet for alleviating pain 8hg/kg group is with rotundine, more then do not have significant difference with the pethidine group.
The influence that table 5 golden red tablet for alleviating pain, rotundine, pethidine react mouse writhing (M ± SD)
Writhing response number of times in group dosage (g/kgd) 15min
NS - (10) 46.5±6.32
Golden red tablet for alleviating pain 4.0 (10) 34.7 ± 13.45*
Golden red tablet for alleviating pain 8.0 (10) 4.3 ± 1.42*** Δ Δs
Rotundine 30mg/kg (10) 13.4 ± 10.32***
Pethidine 20mg/kgd (10) 4.2 ± 1.30***
Δ Δ and rotundine be P＜0.01 relatively
2, to the influence of mice hot plate method: female mice is some, body weight 18-22g, and (to lick metapedes incubation period is the pain index by only being put in screening on 55 ± 0.5 ℃ of hot plates
(6)), surpass 30 seconds (S) incubation period or the happiness leaper rejects), select 40 of qualified mices, survey 1h and 2h behind the ig and lick metapedes incubation period.Result's (table 6) shows the NS group, and there were significant differences with golden red tablet for alleviating pain two dosage groups.Rhizoma Corydalis sheet group and NS group does not more then have significant difference, but itself and golden red tablet for alleviating pain group there were significant differences.
Table 6 golden red tablet for alleviating pain, Rhizoma Corydalis sheet are licked the preclinical influence of metapedes (M ± SD) to mice
Metapedes incubation period (S)
Group dosage (g/kgd)
1h 2h
NS - 20.71±7.14 19.84±6.07
Golden red tablet for alleviating pain 5.0 32.26 ± 9.81* 28.11 ± 8.93* Δ
Golden red tablet for alleviating pain 10.0 36.95 ± 8.18 Δ * * 34.18 ± 6.44* Δ
Rhizoma Corydalis sheet 10.0 25.16 ± 5.17 20.67 ± 4.53
Δ golden red tablet for alleviating pain and Rhizoma Corydalis sheet be P＜0.05 relatively
(5) spasmolytic test-to the active influence of tame rabbit intestinal:
With body weight is 4 of 1.8-2.4kg rabbit (male and female half and half), put to death behind the fasting 16h, cut open the belly and take out small intestinal 2-3 section apart from the about 5cm of duodenum place, every section about 3cm, 18 of the stripped myenteron specimen of preparation, test one by one is after earlier the small intestinal specimen being rinsed with cold tyrode's solution, place the Magnus' bath (volume 20ml, load 1g) of 38.0 ± 0.5 ℃ of isolated organ thermostats.Start one section myenteron shrinkage curve of electrokymograph record
(7)Shrink the adding reagent of stable back Deng myenteron.Promptly with tyrode's solution flushing three times, treat that the myenteron contraction returns to the preceding level of administration and remakes test next time after each test finishes, each specimen is done 3-5 test, and each drug test 6-10 time is tested as follows:
(1) adds golden red tablet for alleviating pain 8 * 10
-2-16 * 10
-3G/ml, myenteron are slowly lax, also obviously strengthen with the increase relexation of dosage, through 6 test myenterons lax to the baseline 0.45 ± 0.2Cm (P＜0.001).Contraction frequency reduces (per minute reduces 2-3 time) slightly, and shrinkage amplitude weakens (amplitude reduces 1-2.0mm) slightly, does 12 tests altogether.
(2) with acetylcholine (ACh) 4 * 10
-3Mg/ml when treating that myenteron shrinks up to peak 4.15 ± 1.9cm, gives 8 * 10
-2Behind the golden red tablet for alleviating pain of g/ml, myenteron is slowly lax, and shrinkage curve reduces by 2.56 ± 0.35cm (P＜0.01).In 10 tests, all antagonism ACh contractions (myenteron is punctured into a straight line) of 4 tests are arranged.
(3) give histamine phosphate 5 * 10
-3Mg/ml, myenteron shrinkage curve give golden red tablet for alleviating pain 2 * 10 up to 2.3 * 0.8cm when being contracted to the peak
-3G/ml, myenteron are lax gradually.Shrinkage curve is reduced to 1.02 ± 0.27cm (P＜0.01).In 8 tests, there are 4 tests all to show to antihistaminic effect (myenteron shrinks in line).
(4) add 5 * 10
-2The barium chloride muscle of hindgut of mg/ml is shunk up to 3.21 ± 0.16cm, and give 4 * 10 this moment
-3-8 * 10
-38 tests of the golden red tablet for alleviating pain of g/ml all do not influence the height (P＞0.05) that myenteron shrinks.
(6) antiinflammatory test
1, on Carrageenan (Carrageenan) causes the bullate influence of rat foot: 32 of rats, body weight 190-220g male and female half and half, 8 every group.Divide four groups, after the special messenger surveyed right back whole mat thickness with micrometer callipers, each group is igNS0.2ml/100g, golden red tablet for alleviating pain 8.0g/kg, 10.0g/kg (capacity 0.2ml/10g) and intramuscular injection (im) hydrocortisone 20mg/kg respectively.Behind im 15min, the ig30min in right back whole pad portion's subcutaneous injection (SC) 1% carrageenin 0.1ml
(8)Survey 1h, 2h, 31 after the administration, 4h, 6h, 8h foot mat thickness, and calculate its swelling value (cause scorching metapedes mat thickness-cause scorching forefoot pad thickness).Golden red tablet for alleviating pain two dosage groups and hydrogen are examined group more all there were significant differences with NS group as a result, and hydrogen is examined group and golden red tablet for alleviating pain group 2-6h no significant difference, but 8h then there were significant differences (table 7).
Table 7 golden red tablet for alleviating pain and hydrogen are examined the influence (X ± SD) that on Carrageenan causes the rat paw edema value
Medicine foot swelling value (mm)
1h 2h 3h 4h 6h 8h
NS 1.60±0.75 2.35±0.79 3.55±0.56 3.08±0.64 3.15±0.67 3.09±0.44
Golden red tablet for alleviating pain I 1.59 ± 0.38 1.69 ± 0.81 2.43 ± 0.73**, 2.04 ± 0.80* 2.90 ± 0.68 2.97 ± 0.53
Golden red tablet for alleviating pain II 1.36 ± 0.52 1.40 ± 0.64*, 2.23 ± 0.81** 2.05 ± 0.63** 2.48 ± 0.80* 2.12 ± 0.62*** Δ
Hydrogen is examined 1.12 ± 0.53 1.32 ± 0.63*, 1.98 ± 0.48**, 1.92 ± 0.60***, 2.41 ± 0.34* 1.57 ± 0.42
Annotate: golden red tablet for alleviating pain I is 8/g/kg, and golden red tablet for alleviating pain 2 is 10g/kg, and it is 20mg/kg that hydrogen is examined.
Δ golden red tablet for alleviating pain and hydrogen are examined relatively P＜0.05 of group
2,5-HT is caused the bullate influence of rat foot:
Male rat, 8 every group, first group of igNS, second group of ig golden red tablet for alleviating pain 10.0g/kg, the 3rd group of ig hydrocortisone 20mg/kg.Press last method in the right back whole pad SC0.01%5-HT0.1ml of portion.
The result shows that the golden red tablet for alleviating pain all has the bullate effect (table 8) of remarkable inhibition rat foot in 1-6h, and effect is not remarkable behind the 8h.
Table 8 golden red tablet for alleviating pain causes the influence (X ± SD) of rat paw edema value to 5-HT
Medicine foot swelling value (mm)
1h 2h 3h 4h 6h 8h
NS 3.23±0.75 2.46±0.34 2.18±0.51 1.94±0.42 1.88±0.56 1.39±0.62
Golden red tablet for alleviating pain 2.41 ± 0.38* 1.81 ± 0.42* 1.20 ± 0.48** 1.04 ± 0.40** 1.00 ± 0.44** 0.90 ± 0.58 Δ
Hydrogen is examined 2.10 ± 0.21*, 1.57 ± 0.37**, 1.18 ± 0.36**, 0.98 ± 0.39**, 0.83 ± 0.40 0.68 ± 0.32**
Δ hydrogen is examined with the golden red tablet for alleviating pain and is compared P＜0.05
3, to the bullate influence of rat granuloma: select 27 of body weight 182 ± 21.5g ♂ rats for use, after the ip pentobarbital sodium 30mg/kg anesthesia, behind the two hand hay cutter of each Mus groin routine disinfection, cut the long osculum of 1cm, the cotton balls that sterilization is weighed as 20mg (adds green grass or young crops, streptomycin mixed liquor 0.2ml, every ml contains penicillin 800mg, streptomycin 650mg implants subcutaneous from otch, behind the skin suture, respectively organize igNS every day, golden red tablet for alleviating pain 5g/kg and 10g/kg, continuously behind the 6d, open otch in the 7th day, cotton balls is taken out in the lump with connective tissue around it, remove fatty tissue, putting oven for drying (spending the night) weighs, weighing deducts the former weight of cotton balls and promptly gets granuloma weight, knows golden red tablet for alleviating pain 10g from table 9, the kg group has remarkable inhibitory action to granuloma induced by implantation of cotton pellets, and the no remarkable inhibitory action of 5g/kg group.
Table 9 golden red tablet for alleviating pain is to the influence of the swollen weight of rat granuloma (M ± SD)
Group dosage (g/kgd) Mus number (only) granuloma heavy (mg)
NS - 9 188.4±59.8
Golden red tablet for alleviating pain 5.0 9 171.5 ± 67.4
Golden red tablet for alleviating pain 10.0 9 89.6 ± 24.5**
(7) to the effect (method of acetic acid) of rat experiment gastric ulcer
(10): 22 of rats, male and female half and half in abdomen state in midair, are divided into two groups down at random, and by only putting into the bell jar that contains ether, take out the anesthesia back, and dorsal position is fixed on the operating-table, cuts off ventral seta, makes conventional skin degerming.In the about 2.5cm of xiphoid-process lower abdomen positive middle part otch, tick duodenum gently, glandular stomach portion is drawn gently outside abdomen, in body of stomach and stomach pyloric antrum intersection, thrust about 0.5mm serous coat under with the microsyringe of interior Sheng 10% acetic acid 0.05ml is flat, the formation pimple is sent stomach back to gently.Sew up stomach wall, flesh layer and skin (during whole surgery, all put stomach animal nose frequently and keep anesthesia usefulness) with the cotton balls that contains ether.Postoperative igNS next day, golden red tablet for alleviating pain 10g/kgd gives 12d continuously, puts to death in 13d, open abdomen, ligation pylorus and cardia are got stomach and are dipped in 1% formalin, fixedly 10min, cut off along greater gastric curvature, stomach is turned up outwell content, wash out food debris gently, observe the ulcer situation of respectively organizing.Ulcer is rounded or oval, central concave, and little all around protuberance, blood capillary gathers.If ulcer heals, protuberance is arranged slightly around then, the surface is ruddy, and radial microgroove is arranged, and represents ulcer index with major diameter of ulcer and minor axis.Ulcer healing percentage rate computing formula:
Relatively there were significant differences with the NS group to know golden red tablet for alleviating pain 10g/kg group from table 10, and the effect that promotes the rat ulcer healing is arranged.
Table 10 golden red tablet for alleviating pain is to the influence of experimental gastric ulcer (method of acetic acid) (M ± SD)
Group dosage number of animals ulcer index healing rate
(g/kgd) (only) (mm) (%)
NS - 9 5.36±1.32
Golden red tablet for alleviating pain 10.0 10 2.47 ± 1.07** 53.92
Brief summary: show that through pharmacodynamics test the golden red tablet for alleviating pain removes has the gastric emptying of delaying; Reduce gastric secretion and total acid output, suppress outside the effect of pepsinia, and have analgesia, spasmolytic, antiinflammatory, antiulcer action.This result shows that the golden red tablet for alleviating pain can regulate gastrointestinal function, plays the effect of depressed liver-energy dispersing and QI regulating and stomach and alleviating pain.
The specific embodiment
Embodiment 1:
Get 1000 raw materials that tablet is required of preparation: Fructus Toosendan 645g, Rhizoma Corydalis (vinegar system) 645g, the anistree leaf 430g of Flos Carthami, Saussurea lappa Clarke 430g.Press following prepared:
1, get Fructus Toosendan, Saussurea lappa Clarke, after the weighing, add the water boil 1.5 hours do not exceed 10 times of amounts, filter, medicinal residues add 8 times of water gagings again and boiled 1.5 hours, filter.Merge 2 times filtrate, it is standby to be concentrated into extractum.
2, get the anistree leaf of Rhizoma Corydalis (vinegar system) and Flos Carthami, after the weighing, added 8 times of amount 50% alcohol reflux 1.5 hours, filter, medicinal residues add 8 times of amount alcohol reflux again and filtered in 1.5 hours, merging filtrate, and it is standby that decompression recycling ethanol gets extractum.
3, the extractum with 1 and 2 steps merge, and drying under reduced pressure becomes dry extract, weigh, and add starch, dextrin is mixed thoroughly, and the back tabletting of granulating obtains 1000 in the tablet of every heavy 0.5 gram, and every contains crude drug 2.15g.
This tablet is oral, a 3-5 sheet, 3-4 time on the one.
Embodiment 2:
The golden red tablet for alleviating pain of the above-mentioned preparation of thin layer chromatography qualitative identification:
(1) get 5 of this product, in the rearmounted conical flask of porphyrize, add petroleum ether (60-90 ℃) 20ml, supersound extraction is 1 hour behind the shake well.Tell the ether layer, put and be concentrated into 0.5ml in the evaporating dish as need testing solution, other gets Radix Aucklandiae control medicinal material powder 0.5g, adds petroleum ether 6ml, and supersound extraction is 1 hour behind the shake well, tells the ether layer, is concentrated into 0.5ml medical material solution in contrast.Test according to thin layer chromatography (35 pages of 1995 editions appendix of Chinese Pharmacopoeia), drawing each 20 μ l of above-mentioned two kinds of solution puts respectively on same silica gel g thin-layer plate, with petroleum ether (60-90 ℃)-ethyl acetate one benzene (14: 3: 3) is developing solvent, launch, taking-up is dried, and spray is with 5% vanillin dilute sulfuric acid solution.Heat after several minutes, in the test sample chromatograph with the corresponding position of control medicinal material chromatograph on, show identical blue spot.
(2) get 15 of this product, porphyrize adds ethanol 35ml shake well, and supersound extraction was filtered after half an hour, and filtrate is flung to ethanol, with 10ml water residue is transferred in the separatory funnel, and 20ml extracts at twice with ethyl acetate, each 10 minutes; Combining extraction liquid, evaporate to dryness.Residue adds 10% hydrochloric acid 10ml hydrolysis in water-bath and filtered the filtrate evaporate to dryness in 2 hours; Residue adds ethanol 1ml dissolving, as need testing solution.Other gets the Quercetin reference substance, and adding ethanol, to make the solution of 2mg/ml be reference substance solution.According to thin layer chromatography (35 pages of 1995 editions appendix of Chinese Pharmacopoeia) test, draw need testing solution 30 μ l and reference substance solution 10 μ l and put respectively on same silica gel g thin-layer plate, be that developing solvent launches with toluene-Ethyl formate-formic acid (5: 4: 1), dry.Natural light is observed down, in the test sample chromatograph with the corresponding position of reference substance chromatograph on, show identical yellow spotting.
(3) get 15 of this product, pulverize and insert in the round-bottomed bottle, add 2ml distilled water and 2ml strong aqua ammonia, moistening medicated powder, add 40ml chloroform (analytical pure) again, reflux, extract, 1 hour, filtration, chloroform is concentrated into 5ml, carry out thin layer chromatography as test liquid.Other gets the tetrahydropalmatine reference substance, adds solution that ethanol makes the suitable 1mg of every 1ml product solution in contrast.Test according to thin layer chromatography (35 pages of 1995 editions appendix of Chinese Pharmacopoeia), draw each 20 μ l of above-mentioned two kinds of solution and put respectively on same silica gel g thin-layer plate, be developing solvent, expansion, take out, dry, spray with bismuth potassium iodide test solution with petroleum ether-chloroform-methanol (10: 6: 1).In the test sample chromatograph with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Embodiment 3Measure the content of tetrahydropalmatine in the golden red tablet for alleviating pain
Get 10 of this product, the accurate title, decide, and porphyrize is put in the conical flask, add ammonia 2ml alkalization, shake up, add benzene 30ml, place filtration in 12 hours, residue adds benzene 30ml again, shake up, placed 12 hours, filter, merging filtrate moves to filtrate in the separatory funnel, the sulphuric acid extraction of usefulness 0.25mol/L three times, each 30ml, merge extractive liquid, adds ammonia and transfers to alkalescence, uses chloroform extraction three times, each 20ml, combined chloroform washs rearmounted water bath method with distilled water 10ml, and residue adds chloroform 5ml dissolving, evaporate to dryness, the dissolving of reuse minimum of chloroform, the accurate sulphuric acid 25ml that adds 0.01mol/L puts water-bath and flings to chloroform, put and be chilled to room temperature, 3 of the red indicators of methylate, with the caustic lye of soda titration of 0.02mol/L, the result proofreaies and correct with blank assay.
Every 1ml 0.01mol/L sulphuric acid is equivalent to 7.1084mg Rhizoma Corydalis second rope.
This product contains total alkaloids, with tetrahydropalmatine (C
21H
26O
4N) calculate, every is no less than 0.5mg.
Claims (11)
1, a kind of compositions for the treatment of stomach pain is characterized in that being made by following materials of weight proportions: Fructus Toosendan 3-5, Rhizoma Corydalis (processed with vinegar) 3-5, the anistree leaf 1-2 of Flos Carthami, Radix Aucklandiae 1-2.
2, compositions according to claim 1 is characterized in that the weight proportion of each raw material is: Fructus Toosendan 3, Rhizoma Corydalis (processed with vinegar) 3, the anistree leaf 2 of Flos Carthami, the Radix Aucklandiae 2.
3, compositions according to claim 1 and 2 is characterized in that the used Radix Aucklandiae is a Saussurea lappa Clarke.
4, a kind of preparation is characterized in that carrying out according to the following steps according to claim 1,2 or 3 described method for compositions: get each raw material in proportion, Fructus Toosendan, Saussurea lappa Clarke are added suitable quantity of water decoct, gained filtrate merges, and it is standby to be concentrated into extractum; Rhizoma Corydalis (processed with vinegar), the anistree leaf of Flos Carthami add an amount of 40-95% ethanol liquid reflux, extract,, and gained filtrate merges, decompression recycling ethanol, and it is standby to be concentrated into extractum; Two step extractum merge, and add conventional excipients, and dry method or wet granulation are made granule, or make capsule, or further tabletting is made tablet.
5, method according to claim 4 is characterized in that raw material water or ethanol extraction 2 times, the 8-10 that extracts used water or ethanol at every turn and be medical material weight doubly, each extraction time is 1-2 hour, extracting the used ethanol of crude drug is 50% ethanol.
6, according to the purposes of the described compositions of arbitrary claim among the claim 1-5 in the medicine of preparation treatment gastric ulcer, stomachache or superficial gastritis.
7, have in preparation according to the described compositions of arbitrary claim among the claim 1-5 and delay gastric emptying, reduce gastric secretion and total acid output effect, suppress the purposes in the medicine of the active at least a effect of pepsinia.
8, the method for the Radix Aucklandiae, Quercetin, tetrahydropalmatine in a kind of qualitative identification compositions comprises a kind of in the following method respectively:
A. it is an amount of to get compositions, and the petroleum ether that adds boiling range and be 60-90 ℃ extracts in right amount, collects extracting solution and prepares need testing solution; It is an amount of that other gets Radix Aucklandiae control medicinal material powder, and the petroleum ether that adds boiling range and be 60-90 ℃ extracts in right amount, collects extracting solution, medical material solution in contrast; Each is an amount of to draw above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, petroleum ether-ethyl acetate one benzene that with the boiling range is 60-90 ℃ is that 14: 3: 3 mixed solvent is developing solvent by volume, launch, spray is with 5% vanillin dilute sulfuric acid solution, heating colour developing, in the test sample chromatograph with the corresponding position of control medicinal material chromatograph on, show identical blue spot;
B. it is an amount of to get compositions, adds ethanol extraction, and extracting solution concentrates, extract obtained with suitable quantity of water with residue suspendible or dissolving, with ethyl acetate extraction, collect acetic acid ethyl acetate extract, remove and desolvate, residue adds 10% hydrochloric acid hydrolysis 1.5-3 hour, filters the filtrate evaporate to dryness; Residue adds dissolve with ethanol, as need testing solution; Other gets the Quercetin reference substance, adds ethanol and makes reference substance solution; Each is an amount of to draw need testing solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with volume proportion is that toluene-Ethyl formate-formic acid mixed solvent of 5: 4: 1 is that developing solvent launches, natural light is observed down, in the test sample chromatograph with the corresponding position of reference substance chromatograph on, show identical yellow spotting;
C. it is an amount of to get compositions, adds a small amount of distilled water and equivalent strong aqua ammonia moistening medicated powder, adds an amount of chloroform extraction again, collects extracting solution, is concentrated into small size, makes test liquid; Other gets the tetrahydropalmatine reference substance, adds ethanol and makes reference substance solution; Each is an amount of to draw above-mentioned two kinds of solution, puts respectively on same silica gel g thin-layer plate, is that petroleum ether-chloroform-methanol of 60-90 ℃ is that developing solvent launches by the mixed solvent of 10: 6: 1 volume ratios with the boiling range, and spray is with bismuth potassium iodide test solution; In the test sample chromatograph with the corresponding position of reference substance chromatograph on, show the speckle of same color;
The above extraction comprises backflow and ultrasonic extraction.
9, the qualitative identification method of the described compositions of arbitrary claim among a kind of claim 1-5, comprise in the following method any one or multiple:
A. it is an amount of to get compositions, and the petroleum ether that adds boiling range and be 60-90 ℃ extracts in right amount, collects extracting solution and prepares need testing solution; It is an amount of that other gets Radix Aucklandiae control medicinal material powder, and the petroleum ether that adds boiling range and be 60-90 ℃ extracts in right amount, collects extracting solution, medical material solution in contrast; Each is an amount of to draw above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, petroleum ether-ethyl acetate one benzene that with the boiling range is 60-90 ℃ is that 14: 3: 3 mixed solvent is developing solvent by volume, launch, spray is with 5% vanillin dilute sulfuric acid solution, heating colour developing, in the test sample chromatograph with the corresponding position of control medicinal material chromatograph on, show identical blue spot;
B. it is an amount of to get compositions, adds ethanol extraction, and extracting solution concentrates, extract obtained with suitable quantity of water with residue suspendible or dissolving, with ethyl acetate extraction, collect acetic acid ethyl acetate extract, remove and desolvate, residue adds 10% hydrochloric acid hydrolysis 1.5-3 hour, filters the filtrate evaporate to dryness; Residue adds dissolve with ethanol, as need testing solution; Other gets the Quercetin reference substance, adds ethanol and makes reference substance solution; Each is an amount of to draw need testing solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with volume proportion is that toluene-Ethyl formate-formic acid mixed solvent of 5: 4: 1 is that developing solvent launches, natural light is observed down, in the test sample chromatograph with the corresponding position of reference substance chromatograph on, show identical yellow spotting;
C. it is an amount of to get compositions, adds a small amount of distilled water and equivalent strong aqua ammonia moistening medicated powder, adds an amount of chloroform extraction again, collects extracting solution, is concentrated into small size, makes test liquid; Other gets the tetrahydropalmatine reference substance, adds ethanol and makes reference substance solution; Each is an amount of to draw above-mentioned two kinds of solution, puts respectively on same silica gel g thin-layer plate, is that petroleum ether-chloroform-methanol of 60-90 ℃ is that developing solvent launches by the mixed solvent of 10: 6: 1 volume ratios with the boiling range, and spray is with bismuth potassium iodide test solution; In the test sample chromatograph with the corresponding position of reference substance chromatograph on, show the speckle of same color;
The above extraction comprises backflow and ultrasonic extraction.
10, the qualitative identification method of a kind of claim 4 or 5 described tablets, comprise in the following method any one or multiple:
A. get 5 in tablet, in the rearmounted conical flask of porphyrize, add boiling range and be 60-90 ℃ petroleum ether 20ml, supersound extraction is 1 hour behind the shake well; Tell the ether layer, put and be concentrated into 0.5ml in the evaporating dish as need testing solution, O.5g other get Radix Aucklandiae control medicinal material powder, adds boiling range and be 60-90 ℃ petroleum ether 6ml, and supersound extraction is 1 hour behind the shake well, tells the ether layer, is concentrated into O.5ml medical material solution in contrast; Draw each 20 μ l of above-mentioned two kinds of solution and put respectively on same silica gel g thin-layer plate, petroleum ether-ethyl acetate one benzene that with the boiling range is 60-90 ℃ is that 14: 3: 3 mixed solvent is developing solvent by volume, launches, and taking-up is dried, and spray is with 5% vanillin dilute sulfuric acid solution; Heat after several minutes, in the test sample chromatograph with the corresponding position of control medicinal material chromatograph on, show identical blue spot;
B. get 15 in tablet, porphyrize adds ethanol 35ml shake well, and supersound extraction was filtered after half an hour, and filtrate is flung to ethanol, with 10ml water residue is transferred in the separatory funnel, and 20ml extracts at twice with ethyl acetate, each 10 minutes; Combining extraction liquid, evaporate to dryness; Residue adds 10% hydrochloric acid 10ml hydrolysis in water-bath and filtered the filtrate evaporate to dryness in 2 hours; Residue adds ethanol 1ml dissolving, as need testing solution; Other gets the Quercetin reference substance, and adding ethanol, to make the solution of 2mg/ml be reference substance solution; Drawing need testing solution 30 μ l and reference substance solution 10 μ l and put respectively on same silica gel g thin-layer plate, is that toluene-Ethyl formate-formic acid mixed solvent of 5: 4: 1 is that developing solvent launches with volume proportion, dries; Natural light is observed down, in the test sample chromatograph with the corresponding position of reference substance chromatograph on, show identical yellow spotting;
C. get 15 of this product, pulverize and insert in the round-bottomed bottle, add 2ml distilled water and 2ml strong aqua ammonia, moistening medicated powder, add the 40ml chloroform again, reflux, extract, 1 hour, filtration, chloroform is concentrated into 5ml, carry out thin layer chromatography as test liquid; Other gets the tetrahydropalmatine reference substance, adds solution that ethanol makes the suitable 1mg of every 1ml product solution in contrast; Drawing each 20 μ l of above-mentioned two kinds of solution and put respectively on same silica gel g thin-layer plate, is that petroleum ether-chloroform-methanol of 60-90 ℃ is developing solvent by the mixed solvent of 10: 6: 1 volume ratios with the boiling range, launches, takes out, dries, sprays with bismuth potassium iodide test solution; In the test sample chromatograph with the corresponding position of reference substance chromatograph on, show the speckle of same color.
11, the method of quality control of the described compositions of arbitrary claim among a kind of claim 1-5, it is characterized in that measuring the wherein content of tetrahydropalmatine, carry out according to the following steps: it is an amount of to get compositions, accurate claim fixed, porphyrize is put in the conical flask, add the ammonia appropriate basesization, shake up, it is an amount of to add benzene, hour filters surplus placing 10, and it is an amount of that residue adds benzene again, shake up, surplus placing 10 hour, filter merging filtrate, filtrate is moved in the separatory funnel, with O.1-0.5mol/L sulphuric acid extraction three times, merge extractive liquid, adds ammonia and transfers to alkalescence, with chloroform extraction for several times, combined chloroform, with the rearmounted water bath method of distilled water wash, residue adds the chloroform dissolving, evaporate to dryness, the dissolving of reuse minimum of chloroform, the accurate sulphuric acid 25ml that adds 0.01mol/L, it is imitative to put water-bath Back stroke dechlorination, put and be chilled to room temperature, 3 of the red indicators of methylate, with the caustic lye of soda titration of 0.02mol/L, the result proofreaies and correct with blank assay.
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| CN104585282A (en) * | 2014-12-31 | 2015-05-06 | 无限极（中国）有限公司 | Biscuit and preparation method thereof |
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| CN104585282A (en) * | 2014-12-31 | 2015-05-06 | 无限极（中国）有限公司 | Biscuit and preparation method thereof |
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