CN1257723C - Powder injection for intravenous injection and its preparing method - Google Patents
Powder injection for intravenous injection and its preparing method Download PDFInfo
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- CN1257723C CN1257723C CN 200410080023 CN200410080023A CN1257723C CN 1257723 C CN1257723 C CN 1257723C CN 200410080023 CN200410080023 CN 200410080023 CN 200410080023 A CN200410080023 A CN 200410080023A CN 1257723 C CN1257723 C CN 1257723C
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Abstract
The present invention discloses a medicine powder for intravenous injection and the preparation method. The medicine is primarily prepared from the raw materials: chlorogenic acid and baicalin. The medicine has obvious curative effect on upper respiratory tract infection, pneumonia and respiratory tract virus infection.
Description
Technical field
The present invention relates to a kind of treatment upper respiratory tract infection, pneumonia, the powder pin of anti-respirovirus.Be the powder pin of feedstock production with the Chinese herbal medicine specifically, relate to the preparation method of this medicine simultaneously.
Background technology
In China, upper respiratory tract infection such as pharyngitis, tracheitis, pneumonia, respiratory virus infection are class commonly encountered diseases frequently-occurring diseases.Not only normal protracted course of disease own, and can cause multiple complications, harm humans health.Along with the development of industrial society, air is by ground contamination in various degree, makes the sickness rate of upper respiratory tract infection such as rhinitis, pharyngitis and pneumonia class disease increase.Though modern medicine thinks that the main pathogenic bacterium of primary disease are streptococcus, antibacterial and virus mixed infection person are much, and diseases such as the acute lymphoblastic inflammation of Yin Faing, rheumatic fever, nephritis have been brought huge misery to the patient thus.Therefore should the disease elimination be very necessary in the early stage.
At present the choice drug of treatment primary disease is an antibiotic, but penicillin medicine has severe anaphylactic reaction, make penicillium sp is have anaphylactoid patient can't to use, and this medicine is strong to antibacterial and virus mixed infection person weak curative effect, drug resistance; It is that SHUANGHUANLIAN transfusion, injectable powder and various silver yellow preparation have dosage forms such as oral liquid, buccal tablets, electuary more widely that the Chinese medicine aspect is used.SHUANGHUANLIAN transfusion, injectable powder curative effect are definite but its effective ingredient is also not too clear and definite, the report of existing at present many anaphylaxiss (" CHINA JOURNAL OF CHINESE MATERIA MEDICA " 1994 754 pages, " modern Application pharmacy " 1994 the 11 the volume first phase 25 pages), hyperemesis (110 pages of the 6th volume second phases of " People's Armed Police's medical science " nineteen ninety-five), hematuria untoward reaction such as (42 pages of the 12nd the 6th phases of volume of " modern Application pharmacy " nineteen ninety-five December).Studies show that in a large number its untoward reaction is mainly caused by phillyrin.Chinese patent 1398626 and 1424084 discloses Yinhuang powder injection for intravenous injection and preparation method thereof and injection silver yellow injectable powder and preparation method thereof.They and the conventional process for purification that extracts of the same employing of other silver yellow transfusion: it is index that Flos Lonicerae is extracted with the chlorogenic acid, and ignored the effect of effective ingredient such as isochlorogenic acid, and chlorogenic acid content is too low, and a large amount of impurity and invalid components cause very big threat to safety, the stability of medication; The leaching process of Radix Scutellariae is an index with the baicalin, though the higher effect of ignoring effective ingredient such as wogonoside of the purity that has has influenced the curative effect of medicine on the contrary.Conventional formulation is used as medicine with extract, is index with chlorogenic acid, baicalin respectively, and active constituent content is low, and composition is indeterminate in the extract, stays hidden danger for safety, the stability of medicine.
Therefore, provide a kind of curative effect reliable, the medicine that toxic and side effects is low presses for.
Summary of the invention
The purpose of this invention is to provide a kind of treatment upper respiratory tract infection powder pin, it is started with from each active component, has studied each component and content range and ratio in the preparation more comprehensively, the active constituent content height, impurity and invalid components content are little, drug safety, stable, and curative effect is reliable.Except that being used for the treatment of upper respiratory tract infection, pneumonia, anti-upper respiratory tract infection also there is better curative effect.Another object of the present invention provides the preparation method of this medicine.
For realizing above purpose, the present invention takes following technical scheme:
The main effective ingredient of powder pin provided by the present invention is grouped into by following one-tenth: chlorogenic acid 10-250mg/g, baicalin 16-400mg/g do not contain phillyrin, jasminoidin.
Main active and concentration are preferably as mentioned above: chlorogenic acid 50-175mg/g, baicalin 100-300mg/g.
Its main active and concentration are optimized for: chlorogenic acid 100-140mg/g, baicalin 170-230mg/g.
The powder pin also comprises the active component of following percentage by weight as mentioned above: isochlorogenic acid 3-80mg/g, wogonoside 2-40mg/g, its concentration is optimized for: isochlorogenic acid 8-50mg/g, wogonoside 4-20mg/g.
For guaranteeing carrying out smoothly of freeze-dry process, increase the rehydration effect of dryed product, in the medicinal liquid we added additive as: sodium chloride, glucose, sorbitol, mannitol, PVP etc. are preferably sodium chloride.The concentration of additive is preferably 10%-15% between 4%-25%, the concentration of glucose sugar is preferably 5%-10%.
Medicine powder pin effective constituent determination method of the present invention is:
The assay method of chlorogenic acid is:
Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica;
With ratio is that 10: 90 acetonitrile-0.4% phosphoric acid solution is a mobile phase
The detection wavelength is 327nm
It is an amount of that the preparation precision of reference substance solution takes by weighing the chlorogenic acid reference substance, adds methanol and make the solution that every 1m1 contains 0.05mg, shakes up, promptly;
This product is got in the preparation of need testing solution, and porphyrize gets that about 0.1g is accurate to be claimed surely, puts in the tool plug conical flask, adds 50% methanol 100ml, claims decide weight, and ultrasonic 30min is put coldly, and weight decided in title again.Supply with 50% methanol and to subtract weight loss, shake up, filter, get that filtrate is an amount of to be added brown measuring bottle and promptly get for test agent solution;
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly;
The assay method of baicalin:
Chromatographic condition and system suitability test octadecyl silane are packing material
With 47: 53: 0.2 methanol-water-phosphoric acid of ratio is mobile phase
Detect wavelength 280nm
It is an amount of that the preparation precision of reference substance solution takes by weighing the baicalin that is dried to constant weight, puts in the 50ml measuring bottle, adds dissolve with methanol, shakes up, and makes the solution that every 1ml contains 60 μ g, promptly;
This product is got in the preparation of need testing solution, porphyrize, and it is fixed to get the accurate title of about 0.1g, puts in the 50ml measuring bottle, adds 50% methanol 50ml jolting, dissolving shakes up, and filters, and precision is measured subsequent filtrate 1ml, puts in the 25ml measuring bottle, add 50% methanol to scale, shake up,, filter with microporous filter membrane (0.45 μ m), that is:
Accurate respectively reference substance solution and the need testing solution 10 μ l of drawing of algoscopy;
The preparation method of medicine powder pin of the present invention is:
(1) chlorogenic acid, the preparation technology of isochlorogenic acid: extracting honeysuckle, 4-20 doubly measures 40%-95% ethanol, reflux, extract, 1-5 time, each 0.5-3 hour, filter, merging filtrate, be evaporated to the extractum shape, it is dry to put into vacuum drying oven, and exsiccant Flos Lonicerae extract is dissolved in and is equivalent to its weight 1-20 and doubly measures in the water of volume, aqueous solution extracts 1-5 time with equal amounts of chloroform, chloroform layer discards (recovery chloroform), and water reuse 0.5-3 doubly measures isoamyl alcohol extraction 1-5 time, merges the isoamyl alcohol layer, concentrating under reduced pressure, vacuum drying gets that to be dissolved in the 40-90% alcoholic solution in right amount an amount of, admixes 200 gram silica gel, water-bath volatilizes (60 ℃), as the column chromatography for separation sample, take by weighing 200-300 order silica gel 2000 grams, with chloroform-methanol (40: 1-2: 1) be solvent, wet method dress post, sample on the dry method is with chloroform-methanol (40: 1-2: 1) eluting, equal-volume collection, every part of 500ml, gradient elution, every 500ml adds methanol 10ml, and TLC follows the trail of inspection, merge same section, through pillar chromatography and in conjunction with recrystallization technology repeatedly, obtain the chemical compound chlorogenic acid, the chemical compound isochlorogenic acid again.
(2) baicalin, the preparation technology of wogonoside: Radix Scutellariae coarse powder, add 2-10 times of water gaging, reflux, extract, 1-5 time, each 0.5--3 hour, filter, merging filtrate, concentrating under reduced pressure, add 0.5-4mol/LHCl solution and transfer PH=0.5～4.0 in right amount, be incubated 0.5-3 hour, placed 8-24 hour, filter, precipitation adds 4-20 times of water gaging, transfer PH=5.0～10.0, add 0.5-2 times of ethanol again, stir and make dissolving, filter, filtrate is transferred pH=1.0～5.5 with HCl solution, is incubated 0.5-3 hour, places 8-24 hour, filter, precipitation is washed till PH=5.0～10.0 with ethanol, and vacuum drying is got above-mentioned dry thing, mix with silica gel (200-300 order), silica gel wet method dress post carries out chromatography, and chloroform-methanol (90: 1～1: 1) is an eluant, and thin layer is checked eluent, equivalent is collected, every part of 500ml is divided into 2 major parts: one (10: 1～90: 1), B (10: 1～1: 1) behind the merging same section, separate out acicular crystal from a part the eluent of chloroform-methanol (10: 1) section, further recrystallization obtains chemical compound 2; B partly passes through silica gel column chromatography (200-300 order), and polydextran gel (LH-20) column chromatography for separation obtains chemical compound 1; Chemical compound 1, baicalin; Chemical compound 2, wogonoside, promptly.
(3) preparation technology of said preparation is: the chlorogenic acid of inventory, baicalin etc. are dissolved in the proper amount of water for injection, and making concentration is chlorogenic acid 0.1%-25mg/ml, baicalin 1-40mg/ml, adds isoosmotic adjusting agent, the pH value of control medicinal liquid.Mend to an amount of with water for injection, regulate the pH value of medicinal liquid, the pH value that makes finished product in 4.0～9.0 scope, quality inspection, fill, lyophilization, packing, promptly.
Among the preparation technology of this chlorogenic acid, isochlorogenic acid employed medical material for the medical material that contains chlorogenic acid, isochlorogenic acid arbitrarily as: Flos Lonicerae, the Cortex Eucommiae etc. are preferably Flos Lonicerae.
The present invention also distinguishes with preparation in the past and embodies in the following areas:
1. improve the purity of effective ingredient, having increases preparation stability, reduces toxicity, reduces dose, is convenient to prepare important function such as modernized preparation.In in the past preparation and technology Flos Lonicerae many be index with the chlorogenic acid, and other compositions such as isochlorogenic acid have been ignored, the purity of chlorogenic acid is also low, and Flos Lonicerae of the present invention is an index with total organic acids and chlorogenic acid how below 20%, has improved the purity of total organic acids when improving curative effect of medication.The purity of traditional handicraft baicalin reaches more than 90%, has but ignored the effect of effective ingredient such as wogonoside, and the present invention is an index with Radix Scutellariae total flavones and baicalin, has improved curative effect of medication.
2. the composition of each component and ratio have close ties to stability of formulation, curative effect etc. in the medicinal substances extract, and we tell component such as chlorogenic acid, isochlorogenic acid in the Flos Lonicerae extract by research, and each components contents ratio is defined; From Radix Scutellariae total flavones, told baicalin, wogonoside, etc. component and defined each components contents by finger printing.Simultaneously components such as chlorogenic acid, isochlorogenic acid, baicalin, wogonoside in the medicine powder pin are done content and defined, improved drug standard, guaranteed safety, stability and the effectiveness of preparation.
3. the suitable pH value of injection can reduce the absorption of medicinal liquid to the untoward reaction of human body, the stability that increases the powder pin, acceleration medicine, so injection must have the suitable substance P H-number.Flavones ingredients such as baicalin are soluble in alkali liquor, and easy crystallization is separated out under the acid condition; Organic acids such as chlorogenic acid are easily molten in methanol, ethanol, 70% ethanol, acetone, dissolve in ethyl acetate, water.We by a large amount of evidences pH value scope of the present invention between 4.0～9.0: when pH value flavones ingredient such as baicalin less than 5.0 time has crystallization to separate out; Chlorogenic acid is the caffeic acid quinate, thereby can also can be by alkali catalyzed hydrolysis by acid, and is very unstable when pH value organic acid composition such as chlorogenic acid greater than 8.0 time, and change color easily takes place.
4. a lot of preparations of being made by chlorogenic acid, baicalin are that numerous medical personnels and patient accept as a kind of common drug for the treatment of upper respiratory tract infection such as pharyngitis, tracheitis.My unit finds that through scrutinizing with long term test the present invention treats pneumonia, preventing respiratory viruses effect in addition, and determined curative effect is worthy of popularization.Chlorogenic acid and isochlorogenic acid have stronger inhibition and killing action to various pathogens and virus.Chlorogenic acid has obvious curative effects to acute laryngopharyngitis and dermatosis, be used for the treatment of clinically acute bacterial catch and put, the leukopenia due to the chemotherapy, chlorogenic acid also has the blood pressure lowering choleretic effect.Studies show that above-mentioned organic acid compositions to various pathogens, has certain inhibitory action as dysentery bacterium, Bacillus typhi, Salmonella paratyphi, streptococcus pneumoniae etc.The external three kinds of antigens to hepatitis B virus (HBV) of baicalin all have significant inhibitory effect; Baicalin does not have the cold service of causing to the normal body temperature rat, and the heating rat is had significant refrigeration function, and is a certain amount of effect relationship; Compositions such as baicalin, wogonoside has the obvious suppression effect to the generation of lipid peroxide, influenza virus PRs there is certain inhibitory action, Cavia porcellus passive cutaneous anaphylaxis, PCA and rat small intestine in vitro, trachea all there is inhibitory action to the anaphylaxis contractile response due to the antigen, can suppress mastocyte and discharge histamine, the activity that can also suppress cyclo-oxygenase and lipoxygenase, synthesizing of blocking-up prostaglandin and leukotriene, thus bring into play antiinflammatory, anti-allergic effects.
In addition, though the present invention has lacked phillyrin than shuanghuanglian powder injection, curative effect does not descend, and raising has all been arranged on the contrary mostly.
The present invention, good effect, toxic and side effects are few.Be the therapeutical effect and the toxic and side effects that show medicine of the present invention, we have done a large amount of experimentatioies, below experimental example be used to further specify the present invention.
Powder of the present invention is at 2, and 2, 4-dinitrophenol causes the refrigeration function of rat fever
One, test objective
Powder pin of the present invention is 7 class new Chinese medicines of Beijing Furuikangzheng Medicine Techn Inst. development, and this test is intended to observe powder of the present invention at chemical irritant-2, and the refrigeration function of rat fever model due to the 2, 4-dinitrophenol is to estimate the analgesic curative effect of this medicine.
Two, test material
1. be subjected to the reagent thing: powder pin of the present invention is provided by Beijing Furuikangzheng Medicine Techn Inst..Clinical plan dosage: 1-2 time/day, 0.4-0.8g/ time, clinical maximum consumption is an X crude drug/Mg).
2. reagent: the chloride injection agent, the 500ml/ bottle is produced lot number: 030415322 by the Beijing Double-Crane Pharmaceutical Co., Ltd; 2,2, 4-dinitrophenol, specification 25g is provided by Chinese Military Medical Science Institute, lot number 860419.
3. animal and feedstuff: the Wistar rat, 102, male, carry out 3 days pre-raisings after buying, body weight 150～200g when being used to test (body weight difference is less than 30g), available from Military Medical Science Institute's animal center, the animal quality certification number: SCXK-(army) 2002-001.The raising condition: 5/cage of group support, standard particle Mus forage feed is freely drunk water, 18～22 ℃ of room temperatures, humidity 50%～70%.The Experimental Establishment quality certification: the moving word of doctor 01-2049 number.
4. equipment: DT-1TB digital electronic clinical thermometer, available from Huachen Medical Meters Co Ltd, Shanghai.
Three, test method
(1) preparation before the test:
1. be in pre-the raising 3 days of environment of same temperature for rat on probation, breadboard temperature is at 17～25 ℃, and in the test all processes, room temperature must not change greater than 3 ℃, avoids noise jamming.
2. duration of test grasps the action of rat at every turn and wants soft, and the degree of depth that clinical thermometer inserts the rat anus is identical with the time, and every rat is clinical thermometer fixedly, fixed test personnel;
3. body temperature prediction: test preceding 3 days with electric body-temperature instrumentation normal body temperature, every day 2 times, continuous 3 days;
4. the rat of using the same day, normal body temperature should be in 36.6～38.3 ℃ scope.Every 30 minutes take temperatures once, measure three times meansigma methods as the normal body temperature of rat.
5.2 the preparation of 2, 4-dinitrophenol solution: precision takes by weighing 2,2, 4-dinitrophenol 300mg places 80ml left and right sides normal saline, and Dropwise 5 mol/LNaOH solution constantly stirs, and treats the clear and bright glassy yellow that becomes of medicinal liquid, and it is standby to 100ml to add the chloride injection agent again.
(2) test method:
1. trial drug
High dose group of the present invention:
Dosage group among the present invention:
Low dose group of the present invention:
SHUANGHUANLIAN matched group: 600mg/10ml/kg;
Model group: 10ml normal saline/kg;
Normal control group: 10ml normal saline/kg.
2. method
After weighing, 102 rats are divided into 6 groups of powder pin low dose group of the present invention, middle dosage group and high dose group, model group, SHUANGHUANLIAN matched group and normal control groups at random.After 0.5 hour, every Mus is in back subcutaneous injection chemistry pyrogen by above-mentioned dosage intravenously administrable for each group--and 2,2, 4-dinitrophenol 33mg/l1ml/kg, intact animal's matched group replaces 2 with the normal saline of equal volume, 2, 4-dinitrophenol.Per 30 minutes take temperatures are 1 time after the pyrogenicity, METHOD FOR CONTINUOUS DETERMINATION 3h.
Four, result of the test
Under this experimental condition, subcutaneous injection chemistry pyrogen-2,2, 4-dinitrophenol, the body temperature of rat begin to raise, and occur the heating peak when 1h, and 3h does not return to basal body temperature yet; Each ratio of powder pin of the present invention and SHUANGHUANLIAN very significantly reduce by 2, and 2, 4-dinitrophenol causes the body temperature (p＜0.001) (result sees table 1 for details) of heating rat
Table 1 powder of the present invention is at 2, the influence of rat fever due to the 2, 4-dinitrophenol
| Group | Body weight/kg | Normal body temperature/℃ | Give 2, fervescence value behind the 2, 4-dinitrophenol/℃ | |||||
| 30min | 60min | 90min | 120min | 150min | 180min | |||
| Dosage group low dose group SHUANGHUANLIAN in the normal group model group high dose group | 37.49±0.40 | 37.49±0.40 37.42±0.29 37.56±0.33 37.60±0.28 37.55±0.35 37.59±0.32 | 0.13±0.29 2.53±0.32 1.91±0.54 1.35±0.59 1.54±0.70 1.22±0.55 | 0.03±0.29 3.17±0.48 1.79±0.43 1.51±0.67 1.95±0.65 1.93±0.78 | 0.00 ± 0.30 2.73 ± 0.39 1.50 ± 0.58 1.12 gives birth to 0.78 1.72 ± 0.64 1.64 ± 0.56 | 0.07±0.36 2.18±0.43 1.34±0.60 0.74±0.68 1.35±0.62 1.30±0.53 | 0.01±0.39 1.88±0.67 1.04±0.55 0.54±0.52 1.02±0.55 0.82±0.62 | -0.03±0.33 1.55±0.58 0.73±0.42 0.43±0.52 0.90±0.43 0.55±0.38 |
Five, conclusion (of pressure testing)
Under this experimental condition, each ratio of powder pin raw material of the present invention all can very significantly reduce by 2, the body temperature of heating rat due to the 2, 4-dinitrophenol; The effect of middle dosage group is best, is better than SHUANGHUANLIAN and other ratio groups.
Powder directed toward bacteria endotoxin of the present invention causes the refrigeration function of rabbit heating
One, test objective
Powder pin of the present invention is provided by Beijing Furuikangzheng Medicine Techn Inst., and this test is intended to observe the refrigeration function of rabbit heating due to the powder directed toward bacteria endotoxin of the present invention, to estimate the analgesic curative effect of this medicine.
Two, test material
1. be subjected to the reagent thing: powder pin of the present invention is provided by Beijing Furuikangzheng Medicine Techn Inst..Clinical plan dosage: 1-2 time/day, 0.4-0.8g/ time, clinical maximum consumption is Xg crude drug/Mml.
2. reagent: the chloride injection agent, the 500ml/ bottle is produced lot number: 030415322 by the Beijing Double-Crane Pharmaceutical Co., Ltd; Bacterial endotoxin is provided by Chinese pharmaceutical biological product institute, tires: every contains 9000EU, lot number 981.
3. animal and feedstuff: 54 of large ear rabbits, male, the quality certification number is provided by Beijing section space animal cultivation center: SCXK capital 2002-005, carried out for 1 week and raise body weight 1.6～2.2kg when being used to test in advance after buying.The raising condition: metal rabbit-hutch list cage is raised, and freely drinks water, and gives standard particle rabbit feedstuff, 18～22 ℃ of room temperatures, humidity 50%～70%.The Experimental Establishment quality certification: the moving word of doctor 01-2049 number.
4. equipment: DT-1TB digital electronic clinical thermometer, available from Huachen Medical Meters Co Ltd, Shanghai.
Three, test method
(1) preparation before the test:
1. be in pre-the raising for 1 week of environment of same temperature for rabbit on probation, breadboard temperature is at 17～25 ℃, and in the test all processes, room temperature must not change greater than 3 ℃, avoids noise jamming;
2. duration of test grasps the action of rabbit at every turn and wants soft, and the degree of depth that clinical thermometer inserts each rabbit anus is identical with the time, and every rabbit is clinical thermometer fixedly, fixed test personnel;
3. body temperature prediction: test preceding 2 days with electric body-temperature instrumentation normal body temperature, every day 2 times, continuous 2 days;
4. the rabbit of using the same day, normal body temperature should be in 38.0～39.6 ℃ scopes, and regular using warming therapy difference must not be above 1 ℃ between each rabbit.Every 30 minutes take temperatures once, the difference of measuring twice, twice body temperature must not surpass 0.2 ℃, with the meansigma methods of this secondary body temperature normal body temperature as this rabbit.
(2) test method:
1. trial drug
High dose group of the present invention:
Dosage group among the present invention:
Low dose group of the present invention:
SHUANGHUANLIAN matched group: 600mg/5ml/kg;
Model group: 5ml normal saline/kg;
Normal control group: 5ml normal saline/kg.
2. method: measure a body temperature every 0.5h before 54 large ear rabbits tests, measure 2 times, the meansigma methods of 2 body temperature is as the normal body temperature of this rabbit.The large ear rabbit that body temperature is qualified is divided into 6 groups of powder pin high dose group of the present invention, middle dosage group and low dose group, SHUANGHUANLIAN group, model group and normal control groups at random, every group 9, each is organized auricular vein and gives warm corresponding medicinal liquid of bathing to 38 ℃ respectively, auricular vein injection endotoxin 1200EU/ml/kg during administration 40min gives 1h, 2h behind the endotoxin, 3h, 4h and 5h and respectively measures the anus temperature one time.
Four, result of the test
Under this experimental condition, behind the intravenous injection endotoxin, rabbit body temperature begins to raise, and occurs the double quotidian fever of exothermal peak when 1h and 3h respectively.Do not return to basal body temperature during 5h yet.When giving endotoxin 1h, SHUANGHUANLIAN, powder pin low dosage of the present invention and high dose have tangible reduction effect to the rising of body temperature, and cooling-down effect be better than in the dosage group, the fervescence value of middle dosage group has the trend of reduction.
When giving endotoxin 2h, compare with model group, SHUANGHUANLIAN, three administration groups of powder pin of the present invention have the reduction effect to the rising of body temperature; Powder pin high dose group animal heat of the present invention has the trend of reduction than SHUANGHUANLIAN group, and the cooling-down effect of the two is better than powder pin low dosage of the present invention and middle dosage.
Give endotoxin 3h, compare with model group, SHUANGHUANLIAN, three administration groups of powder pin of the present invention have the reduction effect to the rising of body temperature; The cooling-down effect of powder pin high dose of the present invention is better than SHUANGHUANLIAN, this medicine low dosage and middle dosage.
Give endotoxin 4h, compare with model group, SHUANGHUANLIAN, three administration groups of powder pin of the present invention have the reduction effect to the rising of body temperature; The cooling-down effect of high dose of the present invention is better than powder pin low dosage of the present invention and middle dosage, and this group fervescence value has the trend of reduction than SHUANGHUANLIAN group.
Give endotoxin 5h, compare with model group, SHUANGHUANLIAN, three administration groups of powder pin of the present invention have the effect of dosage group in the reduction effect powder pin of the present invention to be better than low dose group to the rising of body temperature.(result sees table 2 for details)
Table 2 powder of the present invention is at the influence of rabbit heating due to the endotoxin
| Group | Body weight/kg | Normal body temperature/℃ | Give fervescence value behind the endotoxin/℃ | ||||
| 1h | 2h | 3h | 4h | 5h | |||
| Dosage group low dose group in the model group SHUANGHUANLIAN high dose group | 1.80±0.15 1.77±0.13 1.74±0.11 1.77±0.11 1.77±0.13 | 39.0±0.37 38.8±0.25 38.8±0.23 38.9±0.32 39.0±0.28 | 1.44±0.35 1.09±0.26 1.46±0.19 1.32±0.19 1.33±0.14 | 1.64±0.24 0.77±0.10 1.46±0.11 1.15±0.15 1.17±0.28 | 1.87±0.15 1.31±0.18 1.76±0.53 1.53±0.20 1.67±0.35 | 1.44±0.27 0.84±0.12 1.23±0.52 0.96±0.35 1.59±0.17 | 1.12±0.53 0.53±0.23 0.68±0.40 0.29±0.31 0.71±0.24 |
| Normal group | 1.88±0.26 | 38.8±0.31 | -0.04±0.08 | -0.03±0.11 | -0.02±0.14 | -0.08±0.13 | -0.08±0.14 |
Five, conclusion (of pressure testing)
Under this experimental condition, each group of powder pin of the present invention all has in the refrigeration function dosage group therapeutic effect good to rabbit heating due to the bacterial endotoxin.
Powder of the present invention is at the inhibitory action of caused by dimethylbenzene xylene Mus ear swelling
One, test objective
Powder pin of the present invention is Beijing Furuikangzheng Medicine Techn Inst.'s development new drug, and this test is intended to observe the inhibitory action of powder of the present invention at dimethylbenzene induced mice ear swelling, to estimate the antiinflammatory curative effect of this medicine.
Two, test material
1. animal and raising condition: the KM mice, 100, male, healthy secondary carries out 3 days pre-raisings after buying, body weight 18～22g when being used to test, available from Military Medical Science Institute's animal center, the animal quality certification number: SCXK-(army) 2002-001.The raising condition: 5/cage of group support, standard particle Mus forage feed is freely drunk water, 18～22 ℃ of room temperatures, humidity 50%～70%.The Experimental Establishment quality certification: the moving word of doctor 01-2049 number.
2. instrument: JA5003 electronic balance HANGPING
3. medicine and reagent: powder pin of the present invention is provided clinical plan dosage by Beijing Furuikangzheng Medicine Techn Inst.: 1-2 time/day, and 0.4-0.8g/ time; The chloride injection agent, the 500ml/ bottle is provided by the Beijing Double-Crane Pharmaceutical Co., Ltd, lot number: 030415322; SHUANGHUANGLIAN FENZHENJI, the 600mg/ ampoule is produced lot number 0306225 by No. 2 TCM Factory; Dimethylbenzene.
Three, test method
1. trial drug:
High dose group of the present invention:
Dosage group among the present invention:
Low dose group of the present invention:
SHUANGHUANLIAN matched group: 900mg/20ml/kg;
Model group: 20ml normal saline/kg;
2. test method:
100 of male KM mices are divided into 5 groups of powder pin low dose group of the present invention, middle dosage group and high dose group, model group and SHUANGHUANLIAN matched groups, 20 every group at random by body weight.By each intravenously administrable of upper and lower noon of above-mentioned dosage once, be administered once morning next day, and administration is 3 times altogether.Drip 50ul dimethylbenzene in auris dextra during last administration 30min, left side ear is left intact, dislocation is put to death behind the 15min, cut behind the ears with 7mm diameter card punch respectively about same position is laid behind the two circle auricles along auricle, electronic balance claims the auricle weight in wet base, every Mus is the swelling degree with the weight that the weight of auris dextra sheet deducts left auricle, and calculates inhibitory rate of intumesce.
Suppression ratio=(model group swelling degree-medication group swelling degree)/model group swelling degree * 100%
Four, result of the test
Originally discover: compare with model group, each group of SHUANGHUANLIAN and powder pin of the present invention can obviously suppress the mice ear that dimethylbenzene causes; The results are shown in Table 3.
Table 3 powder of the present invention is at the inhibitory action (n=20) of Mus ear swelling
| Body weight/g | Swelling degree/g | Inhibitory rate of intumesce % | |
| Dosage group high dose group of the present invention among the low dose group the present invention of the present invention of model group SHUANGHUANLIAN group | 16.3±6.92 7.7±6.55 8.9±4.28 10.7±3.15 12.7±5.28 | 0 52.8 45.4 34.4 22.1 |
Five, conclusion (of pressure testing)
Under this experimental condition, powder of the present invention has inhibitory action at the Mus ear swelling that dimethylbenzene causes, and inhibition degree and dosage are inverse ratio.
The anti-diseases of respiratory system toxic action of powder pin of the present invention
One, test objective
Powder pin of the present invention is Beijing Furuikangzheng Medicine Techn Inst.'s development new drug, and this test is intended to observe the inhibitory action of powder of the present invention at each Strain, to estimate the antiviral curative effect of this medicine.
Two, test material
1. animal and raising condition: the animal Kunming mouse, male and female half and half, 18～20g, 4 ages in week are available from Military Medical Science Institute's animal center, the animal quality certification number: SCXK-(army) 2002-001.The raising condition: 5/cage of group support, standard particle Mus forage feed is freely drunk water, 18～22 ℃ of room temperatures, humidity 50%～70%.The Experimental Establishment quality certification: the moving word of doctor 01-2049 number.
2. reagent: viral respiratory syncytial virus (RSV) Long strain is derived from U.S. ATCC strain library, lot number: 990521; Adenovirus Ad3V, Virology Inst., Chinese Academy of Preventive Medical Science preserves.Influenza A1 virus (A1V), Mus pneumonopathy venom is preserved by Virology Inst., Chinese Academy of Preventive Medical Science.SHUANGHUANGLIAN FENZHENJI, the 600mg/ ampoule is produced lot number 0306225 by No. 2 TCM Factory; Culture medium: IMDM (U.S. GIBC product); Tetrazolium bromide (MTT) is available from magnificent company.Eagle ' s keeps liquid, is provided by Virology Inst., Chinese Academy of Preventive Medical Science.
3. be subjected to the reagent thing: powder pin of the present invention, by Beijing Furuikangzheng Medicine Techn Inst..
4. instrument: instrument enzyme mark analyzer, LBK ultraviolet-uisible spectrophotometer produce by the U.S., the LC-9A infusion pump, and refrigerated centrifuge, Olympus phase contrast microscope are day island proper Tianjin company product.
Three, test method
1. the screening of cell and preparation: used 3 kinds of viruses are inoculated in VERO, HEP2 cell respectively to measure sensitivity, and cell goes down to posterity according to a conventional method.Make 2 * 10-5/mL cell suspension simultaneously, every hole 01mL, it is standby to form cell monolayer behind the 24h.
2. primary dcreening operation and grouping experiment: form the cell monolayer hole and discard culture fluid, the drug effect 24h that adds pair cell avirulence concentration, the sucking-off medicinal liquid adds viral liquid, by the time of general virus absorption, all adopt 37 ℃ of absorption 1h sucking-off, add the liquid of keeping of pastille, put 37 ℃, 5%CO2 calorstat cultivation 48～96h, treat that virus control hole pathological changes reaches ++ +～++ ++, the result is just often surveyed in the cell contrast.Divide 4 groups with the significant virus of primary dcreening operation and experimentize, inquire into the concrete effect link of medicine.High dose: 24h administration before infecting, the sucking-off medicinal liquid is washed 3 times with Hank ' s liquid, adds viral liquid.In the dosage group: infect the back administration, first liquid feeding is put 37 ℃ of absorption 1h flush away viruses and is added pastille and keep liquid.Low dose group: medicinal liquid and virus add cellular layer simultaneously.IV group: medicinal liquid adds cellular layer with viral the mixing after putting 37 ℃, 2h.Below all establish normal control and virus control.
3. the foundation of infected animal model and dosage regimen: viral pneumonia modelling
[1]Laboratory animal is divided into viral infection model group, normal control group, antiviral variable concentrations administration group at random, the positive control drug comparable group, every group 10, all raise under the same conditions, be intravenously administrable, dosage is as follows: powder pin of the present invention and positive control drug SHUANGHUANGLIAN FENZHENJI are 320,160,80,40mg/k g.
Four, result
Utilize light microscopy checking cytopathy (CPE), the result shows that powder pin of the present invention can suppress above-mentioned virus to some extent to histiocytic CPE, sees Table 4.
Table 4 powder of the present invention causes the cytopathy inhibitory action at virus
| Virus | The cell contrast | Virus control | Test |
| RSV AD3V A1V | - - - | ++++ ++++ ++++ | - - - |
Group result is observed on the CPE basis under mirror, uses the MTT staining cell, measures absorbance, so that further reflect the difference of dosing and not dosing objectively.High dose: can experiment purpose be to observe medicine enter cell or be adsorbed in cell surface, to stop viral absorption and to penetrate.The result shows, this method of powder pin of the present invention is to RSV, Ad3V, A1V3 kind virus meaningful (P＜0 05).Middle dosage group: can experiment purpose be to observe medicine work to entering intracellular virus, suppresses its biosynthesis and ripe release.The result shows that this method of powder pin of the present invention is to 3 kinds of viruses meaningful (P＜0 05).III, IV group: for medicine and virus act under different condition, observe medicine to the direct deactivation of virus, the result shows that powder of the present invention all has effect at 3 kinds of viruses.Illustrate powder pin of the present invention no matter be medicine and virus add simultaneously cellular layer or medicine and virus earlier effect 2h add cellular layer then, all have antiviral effect.The protective rate of pair cell reaches as high as 96%, sees Table 5.
Table 5 powder medicine administered by injection of the present invention thing is to the direct deactivation of virus
| NO. | RSV | AD3V | A1V | P |
| I II III IV | 91.6 90.1 88.9 66.4 | 84.3 84.7 79.2 80.2 | 94.1 94.6 80.5 73.4 | ＜0.05 ＜0.05 ＜0.05 ＜0.05 |
Viral inflammation due to histopathological examination powder of the present invention infects at FM1 has significant protective effect, and treatment group Mus lung tissue structure base is normal, and there are a large amount of pathological changes in virus control group Mus pulmonary, and medicine calculates with following formula the histiocyte protective rate.Protective rate (%)=dosing hole A value-virus control hole A value/cell control well A value * 100%.See Table 6.
The protective effect of the viral pneumonia due to table 6 powder of the present invention infects at FM1
| NO | Ratio | Index ( x±s) | Protective rate (%) |
| Virus control group normal control group API | - - | 1.785±0.312 0.910±0.103 1.401±0.171 1.232±0.069 1.199±0.062 | - - 30.73 41.42 48.68 |
Five, conclusion (of pressure testing)
Under this experimental condition, the antivirus action of powder pin of the present invention is not only confirmed on cellular level, can suppress, delay cytopathy significantly occurs, also show on the integral level, can effectively suppress the generation and the development of respiratory-tract viral pneumonia such as influenza virus, curative effect is preferably arranged.About the antiviral mechanism of action of powder pin of the present invention, from the grouping experiment result as can be seen, effect is multipath, and the effect of direct inactivation of viruses is not only arranged, and to being adsorbed in cell surface and entering intracellular virus inhibitory action is arranged also.
Powder pin pyrogen test of the present invention
One, test objective
Whether powder pin of the present invention is provided by Beijing Furuikangzheng Medicine Techn Inst., in the medicinal liquid vein injection large ear rabbit body with doses, at the appointed time, observes the situation of rabbit fervescence, up to specification with the limit of judging contained pyrogen in the test sample.
Two, test material
(1) be subjected to the reagent thing: powder pin of the present invention is provided by Beijing Furuikangzheng Medicine Techn Inst..Clinical plan dosage: 1-2 time/day, 0.4-0.8g/ time, clinical maximum consumption is Xg crude drug/Mml/60kg body weight.
(2) instrument and reagent: the chloride injection agent, the 500ml/ bottle is produced lot number: 030415322 by the Beijing Double-Crane Pharmaceutical Co., Ltd; The DT-1TB electronic clinical thermometer, Huachen Medical Meters Co Ltd, Shanghai produces.
(3) animal and feedstuff: large ear rabbit, male, the quality certification number is provided by Beijing section space animal cultivation center: SCXK capital 2002-005, carry out a week and raise body weight 2.3～2.4kg when being used to test in advance after buying.Raising condition: raise with metal rabbit-hutch list cage, freely drink water, give granule rabbit feedstuff, humidity 50%～70%, 18～22 ℃ of temperature.The Experimental Establishment quality certification: the moving word of doctor 01-2049 number.
Three, test method
(1) preparation before the test:
1. the environment that is in same temperature for rabbit on probation before the pyrogen test is raised in advance, and breadboard temperature is at 17～25 ℃, and in the test all processes, room temperature must not change greater than 3 ℃, avoids noise jamming;
2. water is can't help in rabbit fasting before test, and it is soft that operation is wanted;
3. to insert the degree of depth of each rabbit anus identical with the time for clinical thermometer;
4. every 30 minutes take temperatures once, measure secondary, the difference of secondary body temperature must not surpass 0.2 ℃, with the meansigma methods of this secondary body temperature normal body temperature as this rabbit;
5. the rabbit of using the same day, normal body temperature should be in 38.0～39.6 ℃ scopes, and regular using warming therapy difference must not be above 1 ℃ between each rabbit.
(2) selecting of animal: the condition of selecting is identical when checking test sample, and only liquid medicine injection not every 30 minutes take temperatures once, is measured four times, and the difference of four body temperature must not be above 0.4 ℃.
(3) inspection technique; Get 3 of suitable large ear rabbits, measure after its normal body temperature in 15 minutes, slowly inject from auricular vein with aseptic manipulation and to be warmed to 38 ℃ powder pin of the present invention, measure its body temperature once every 30min by preceding method then, survey altogether six times, with the highest normal body temperature that once deducts in six body temperature, be this rabbit fervescence the number of degrees (℃), if body temperature is lower than normal body temperature after the administration, then the fervescence number of degrees are in " 0 ".
Four, the result judges
(1) judges retrial
1. as in 3 rabbits 1 fervescence is arranged more than 0.6 ℃ or 0.6 ℃;
2.3 only the rabbit fervescence all is lower than 0.6 ℃, but the sum that raises reaches and occur one of above-mentioned situation more than 1.4 ℃ or 1.4 ℃ and should get 5 rabbit retrials in addition, inspection method is the same.
(2) nonpyrogenic
1. in 3 of preliminary examination rabbits, fervescence all is lower than 0.6 ℃, and 3 rabbit fervescence sums are lower than 1.4 ℃;
2. in 5 of retrial rabbits, the rabbit number of fervescence more than 0.6 ℃ or 0.6 ℃ only has 1, and the fervescence that preliminary examination, retrial merge 8 rabbits adds up to below 3.5 ℃ or 3.5 ℃.
Meet one of above-mentioned situation and think that all the pyrogen test of test sample is up to specification.
(3) pyrogen test is defective
1. in 3 rabbits of preliminary examination, the rabbit number of fervescence more than 0.6 ℃ or 0.6 ℃ is above 1;
2. in 5 rabbits of retrial, the rabbit number of fervescence more than 0.6 ℃ or 0.6 ℃ is above 1;
3. merge the fervescence sum of 8 large ear rabbits above 3.5 ℃ in preliminary examination and retrial.
One of above-mentioned situation occurs and think that all the pyrogen test of test sample is against regulation.
Five, result of the test
Vein gives the powder pin of the present invention that tries, and 2 rabbit body temperatures do not raise, and 1 fervescence is lower than 0.6 ℃ for 0.1 ℃, and 3 fervescence sums (0.1 ℃) are lower than 1.4 ℃, the results are shown in Table 7.
Table 7 powder pin of the present invention pyrogen test result
| Count | Body weight (kg) | Dosage (the mg crude drug/kg) | Normal body temperature (℃) | High fever after the administration (℃) | Fervescence (℃) |
| 1 3 5 | 2.4 2.3 2.4 | X X X | 38.5 38.9 38.9 | 38.6 38.8 38.8 | 0.1 0 0 |
Six, conclusion (of pressure testing)
Under this experimental condition, the limit of the contained pyrogen of powder pin of the present invention is up to specification.
Powder pin undue toxicity of the present invention checks
One, test objective
The medicinal liquid vein of the present invention of doses is injected in the mice body, at the appointed time, observe the death condition of mice, whether up to specification with the undue toxicity who judges test sample.
Two, test material
(1) be subjected to the reagent thing: powder pin of the present invention is provided by Beijing Furuikangzheng Medicine Techn Inst..Clinical plan dosage: 1-2 time/day, 0.2-0.4g/ time, clinical maximum consumption is Xg crude drug/Mml/60kg body weight.
(2) animal and feedstuff: 5 KM mices, male, healthy secondary carries out 3 days pre-raisings after buying, body weight 17～20g when being used to test, available from Military Medical Science Institute's animal center, the animal quality certification number: SCXK-(army) 2002-001.The raising condition: 5/cage group support, standard feed is fed, and freely drinks water 18～25 ℃ of room temperatures, humidity 50%～70%, illumination light and shade each 12 hours, the Experimental Establishment quality certification: the moving word of doctor 01-2049 number.Full nutrition granule Mus feedstuff is by Military Medical Science Institute's animal center development.
Three, test method
5 of mices by intravenous administration 0.5ml/ are only weighed the back, and normal the raising must not be dead in 48 hours, weighs.
Four, the result judges
(1) judges retrial
When in 5 mices death condition being arranged, should get 10 retrials of mice of body weight 18～19g in addition, inspection method is the same.
(2) undue toxicity's passed examination
1. 5 of preliminary examination mices did not have dead in 48 hours;
2. 10 of retrial mices did not have dead in 48 hours.
Meeting one of above-mentioned situation, to think that all the undue toxicity of test sample checks up to specification.
(3) undue toxicity checks defective
5 mices of preliminary examination had death in 48 hours; 10 mices of retrial had death in 48 hours.
Meeting above-mentioned situation, to think that the undue toxicity of test sample checks against regulation.
Five, result of the test
All mice does not have dead after administration in 48 hours.
Six, conclusion (of pressure testing)
Under this experimental condition, powder pin undue toxicity of the present invention is up to specification.
Powder pin acute toxicity test in mice of the present invention
One. test objective
The toxicity of powder pin of the present invention is lower, surveys the median lethal dose(LD 50) (LD that do not come out
50), observe the single vein and irritate stomach giving mice acute toxic reaction and death condition that powder pin of the present invention is produced, draw the maximum tolerated dose of iv and ig administration, its safety is estimated, provide reference for clinical.
Two. material
(1) tried thing
1. be subjected to the reagent thing: powder pin of the present invention is provided by Beijing Furuikangzheng Medicine Techn Inst..Clinical plan dosage: 1-2 time/day, 0.4-0.8g/ time, clinical maximum consumption is the Dg crude drug, i.e. Fg crude drug/60kg body weight.
2. reagent: distilled water, available from Chinese People's Liberation Army General Hospital.
(2) animal
60 KM mices, male and female half and half, healthy secondary carries out 3 days pre-raisings after buying, body weight 18～22g when being used to test, available from Military Medical Science Institute's animal center, the animal quality certification number: SCXK-(army) 2002-001.The raising condition: 5/cage group support, standard feed is fed, and freely drinks water 18～25 ℃ of room temperatures, humidity 50%～70%, illumination light and shade each 12 hours, the Experimental Establishment quality certification: the moving word of doctor 01-2049 number.Full nutrition granule Mus feedstuff is by Military Medical Science Institute's animal center development.
Three. test method
(1) dosage determines
The concentration Ag crude drug/ml of used powder pin of the present invention, according to mice single vein (iv) the administration heap(ed) capacity be aml/bg, the dosage of determining mice iv is vg crude drug/kg; It is dml/10g that the mice single is irritated stomach (ig) administration heap(ed) capacity, and the dosage of determining mice ig is fg crude drug/kg.
(2) medication
1. route of administration: iv is consistent with the approach of clinical vein administration.
2. administration capacity: mice iv vml/10g; Ig dml/10g.
3. for reagent: medicine is provided by the Drug Manufacturing Room of this institute.
(3) test basis
Method requires and these section office formulate " pharmacological testing standard operating procedure " according to " acute toxicity test " in bureau of drug administration of Ministry of Health of the People's Republic of China " study of tcm new drug guide " and " new drug (Western medicine) the preclinical study guideline compilation " (SOP) carries out.
(4) method
60 of KM mices are divided into iv group of the present invention, ig of the present invention group and three groups of matched groups at random according to body weight, and 20 every group, male and female half and half, fasting be can't help water 12 hours before the administration, and iv group mice of the present invention is according to the administration capacity intravenously administrable of 0.25ml/10g; Ig group mice of the present invention is according to the administration capacity gastric infusion of 0.4ml/10g.At once observe the reaction of animal after the administration, form, behavioral activity, the mental status, appetite, defecation and the color thereof, fur, nose, eye, the mouth that comprise animal have or not abnormal secretion thing and animal dead situation, the dead timely postmortem of animal is if there is macroscopic pathological changes then to carry out histopathologic examination.Administration was observed once in the 1st day per 1 hour, and observe once later every day, observed reaction of record animal toxicity and death condition continuously 7 days.The weighing mice before test, the body weight of test back the 3rd day and the 7th day.Response situation overall merit result of the test from body weight and animal obtains the maximum tolerated dose of powder of the present invention at mice single vein and gastric infusion.If death occurs, then carry out acute median lethal dose(LD 50) (LD
50) mensuration.
Four. result of the test
(1) MTD of iv
KM mice single vein gives the powder pin 17.075g crude drug/kg of the present invention that tries, be equivalent to 187.5 times of the clinical maximum consumption per day of people, at once the righting reflex loss of animal after the administration, righting reflex recovers in the palpitating speed, 3min, heartbeating recovery is normal, the no abnormal performance of animal does not occur any unusually during the whole test, body weight increases with the prolongation of test period, with concurrent control group zero difference (p＞0.05), duration of test is not seen animal dead.Put to death mice when the observation period finishes, postmortem does not have macroscopic variation.
(2) MTD of ig
KM mice single is irritated stomach and is given the powder pin 27.320g crude drug/kg of the present invention that tries, be equivalent to 300 times of the clinical maximum consumption per day of people, at once reach after the administration do not occur during the whole test any unusual, body weight increases with the prolongation of test period, with concurrent control group zero difference (p＞0.05), duration of test is not seen animal dead.Put to death mice when the observation period finishes, postmortem does not have macroscopic variation.
Five. conclusion (of pressure testing)
Under this experimental condition, powder pin of the present invention is given the maximum tolerated dose＞Ag crude drug/kg of mice single intravenously administrable, is equivalent to 187.5 times of the clinical maximum consumption per day of people; Single is irritated the maximum tolerated dose＞Sg crude drug/kg of stomach, is equivalent to 300 times of the clinical maximum consumption per day of people, and the clinical consumption of this medicine of this test prompting is a quite safe dosage.
Powder pin blood vessel irritation test of the present invention
One, test objective
This test is intended to observe this product continuous auricular vein administration every day, rabbit ear edge vein is had or not the vascular stimulation effect, for clinical drug safety provides reference.
Two, test material
1. be subjected to the reagent thing: powder pin of the present invention is provided by Beijing Furuikangzheng Medicine Techn Inst..Clinical plan dosage: 1-2 time/day, 0.4-0.8g/ time.
2. reagent: the chloride injection agent, the 500ml/ bottle is produced lot number: 030415322 by the Beijing Double-Crane Pharmaceutical Co., Ltd.
3. animal and feedstuff: large ear rabbit, male, the quality certification number is provided by Beijing section space animal cultivation center: SCXK capital 2002-005, carried out for 1 week and raise body weight 2.2～2.6kg when being used to test in advance after buying.The raising condition: metal rabbit-hutch list cage is raised, and freely drinks water, and gives standard particle rabbit feedstuff, 18～22 ℃ of room temperatures, humidity 50%～70%.The Experimental Establishment quality certification: the moving word of doctor 01-2049 number.
Three, test method
1. get health, the ear edge does not have 6 of the large ear rabbits of wound, is divided into two groups of powder pin group of the present invention and chloride injection agent matched groups at random, 3 every group.The administration group is with aseptic manipulation auricular vein drug administration by injection 91.07mg crude drug/0.67ml/kg, matched group gives with the agent of volume chloride injection, control injection speed 3ml/min, 9:00am respectively organizes rabbit left side auricular vein drug administration by injection once respectively at every day, successive administration 3d.
2. get health, the ear edge does not have 6 of the large ear rabbits of wound, is divided into two groups of powder pin group of the present invention and chloride injection agent matched groups at random, 3 every group.The administration group is with aseptic manipulation auricular vein drug administration by injection mg/0.67ml/kg, and matched group gives with the agent of volume chloride injection, control injection speed 3ml/min, and 9:00am respectively organizes rabbit left side auricular vein drug administration by injection once respectively at every day, successive administration 7d.
Four, observation index
1. after administration every day, the red and swollen situation of partial vein blood vessel of perusal administration and surrounding tissue.
2. 24h knocks head execution with animal after the last administration, respectively at injection site proximal part 1.5cm to 3cm place clip auricular vein, fix with 10% formalin solution, routine pathology section HE dyeing, observation has or not tissue degeneratiaon or irritant reaction such as necrosis, thrombosis and endothelial injury.
Five, result of the test
Powder pin auricular vein drug administration by injection 91.07mg crude drug/0.67ml/kg of the present invention, 3d and 7d have expansion, hemorrhage effect to rabbit ear local vascular continuously, and cell infiltration, do not see vascular wall degeneration, downright bad damage and thrombosis, administration changed slightly heavy than 3 days blood vessels of administration in 7 days.This medicament intravascular stimulates pathological changes to belong to reversibility, slight vascular lesion generally.
Six, conclusion (of pressure testing)
Under this experimental condition, powder pin of the present invention does not produce degeneration, necrosis and the thrombosis of blood vessel wall.
The outer hemolytic test of powder needle body of the present invention
One, test objective
This test is intended to observe powder of the present invention and has or not haemolysis and cause agglutination at rabbit erythrocyte.
Two, test material
1. be subjected to the reagent thing: the same
2. reagent: the chloride injection agent, the 500ml/ bottle is produced lot number: 030415322 by the Beijing Double-Crane Pharmaceutical Co., Ltd
3. instrument: 800B type desk centrifuge, Anting Scientific Instrument Factory, Shanghai produces; The SY21-Ni electric-heated thermostatic water bath, mayor of Beijing bearing instruments and meters company produces.
Three, test method
The preparation of 1.2% red blood cell suspension: rabbit heart is got blood 20ml, put into fill bead triangular flask gently jolting 10 minutes remove and defibrinate, make into defibrinated blood, the normal saline that adds 10 times of amounts shakes up, centrifugal 3min (rotating speed 2000rpm/min), remove supernatant, sedimentary erythrocyte reuse normal saline washs till the supernatant water white transparency.It is standby that the gained erythrocyte is made into 2% suspension with normal saline.
2. test method: get 7 in test tube, add 2% red blood cell suspension and normal saline successively by table 8, bathe half an hour in 37 ℃ of temperature behind the mixing, (the 6th manages not add and is subjected to test product, as the blank group to add not commensurability medicinal liquid by table 8 then; The 7th pipe does not add and is subjected to test product, replaces normal saline with distilled water, as positive controls), each pipe is shaken up rearmounted 37 ℃ of water bath heat preservation 4h gently.Beginning was observed once every 15 minutes, after 1 hour, observed once observation haemolysis as shown in table 9 and agglutination phenomenon every 1 hour.
The outer hemolytic test application of sample table of table 8 powder needle body of the present invention
| The pipe number | 1 | 2 | 3 | 4 | 5 | 6 | 7 |
| 2% red cell suspension (ml) normal saline (ml) distilled water (ml) is subjected to test product (ml) | 2.5 2.O - O.5 | 2.5 2.1 - 0.4 | 2.5 2.2 - 0.3 | 2.5 2.3 - 0.2 | 2.5 2.4 - 0.1 | 2.5 2.5 - - | 2.5 - 2.5 - |
Table 9 erythrocyte hemolysis, coagulation criterion
| Full haemolysis part haemolysis does not have the haemolysis coagulation | The clear and bright redness of solution, the pipe end, is acellular residual; The clear and bright redness of solution or brown, the pipe end, have a small amount of erythrocyte residual; Erythrocyte all sinks, the supernatant liquid achromatism and clarity; The erythrocyte aggregation in bulk can not disperse after the jolting; |
Four, the result judges
1. with 0.3ml powder pin (the 3rd pipe), but the person that do not produce the haemolysis in 2 hours thinks injection.
2. if any the phenomenon of red blood cell condensation, can further judge it is true cohesion or pseudo agglutination by purgation.If condensation product again can homodisperse after test tube vibration, or aggregation is placed on the microscope slide, drips 2 normal saline at the coverslip edge, microscopically is observed, and the cohesion erythrocyte can be pseudo agglutination by the person of breaking up, and test sample can supply clinical practice.If condensation product is not shaken diffusing or be not true cohesion by the person of breaking up on slide, test sample should not be for clinical use.
Five, result of the test
Haemolysis does not appear in the 1-6 pipe 4h; The 1-4 pipe coacervation occurs at different time points, and the erythrocyte homodisperse belongs to pseudo agglutination after the jolting; 7 pipe complete hemolysis.(the results are shown in Table 10)
The outer hemolytic test result of table 10 powder needle body of the present invention
| Time | 1 | 2 | 3 | 4 | 5 | 6 | 7 |
| 15min 30min 45min 1h 2h 3h 4h | - - - * ** *** *** | - - - * ** *** *** | - - - - ** *** *** | - - - - *** *** *** | - - - - *** *** *** | - - - - - - - | + + + + + + + |
Remarks: the no haemolysis of "-" expression; "+" expression complete hemolysis; " ± " expression part haemolysis; " * " represents pseudo agglutination
Six, conclusion (of pressure testing)
Under this experimental condition, the powder of the present invention that tries does not have haemolysis at rabbit erythrocyte, has pseudo agglutination phenomenon in various degree to take place.
Powder pin systemic allergy test of the present invention
One, test objective
Powder pin of the present invention is the new Chinese medicine of Beijing Furuikangzheng Medicine Techn Inst.'s development, and this test is intended to investigate powder pin of the present invention and has or not the whole body sensitization, is reference data for clinical drug use.
Two, test material
1. be subjected to the reagent thing: the same
2. reagent: the chloride injection agent, the 500ml/ bottle is produced lot number: 030415322 by the Beijing Double-Crane Pharmaceutical Co., Ltd; Fresh ovalbumin is mixed with 10% concentration with normal saline and is for experiment.
3. animal: albino guinea-pig, male and female half and half are provided by Beijing section space animal cultivation center, the quality certification number: SCXK capital 2002-005, carry out a week and raise body weight 270～350g when being used to test in advance after buying.The raising condition: 9/cage of group support, granule Cavia porcellus forage feed is freely taken the photograph water, 20～22 ℃ of room temperatures, humidity 50%～70%.The Experimental Establishment quality certification: the moving word of doctor 01-2049 number.
Four, test method
1. grouping: 18 of healthy guinea pigs are divided into three groups of powder pin group of the present invention, ovalbumin positive controls and chloride injection agent negative control group, 6 every group, male and female half and half at random by body weight.
2. animal subject sensitization: each treated animal by the sterile working next day respectively lumbar injection only give powder pin of the present invention, 10% ovalbumin and chloride injection agent 0.5ml/, sensitization is three times altogether.
3. attack: each treated animal is divided into 2 batches, 3 every batch, a collection of after sensitization first 14d respectively the corresponding medicinal liquid 1ml/ of intravenous injection only attack; Another batch corresponding medicinal liquid 1ml/ of 21d intravenous injection after sensitization first only attacks.
Five, observation index
After attacking administration, in the 15min, observe animal and have or not cough, grab nose, erect situations such as hair, dyspnea, spasm, shock and death, and press the listed standard scoring of table 11, marked 〉=2 o'clock, be judged to be the hypersensitive test positive.(the results are shown in Table 11)
Table 11 systemic anaphylaxis reaction standards of grading
| Scoring | Sign |
| 0 1 2 3 4 | No significant reaction is slightly grabbed nose, is trembled or perpendicular hair appearance cough, repeatedly grab nose, tremble or perpendicular hair repeatedly or cough continuously, with dyspnea or spasm, tic spasm, tic, gatism, shock death |
Six, result of the test
Occur spasm, tic, gatism, last dead in 6 Cavia porcellus 15min of positive group, marked 4 fens, be the anaphylaxis positive; Negative group and 6 Cavia porcelluss of powder pin group of the present invention do not have significant reaction, mark 0 fen, are the anaphylaxis feminine gender.(the results are shown in Table 12)
Table 12 powder pin of the present invention systemic allergy test result
| Powder pin group | Positive group | Negative group | ||||||
| Animal number | Sex | Scoring | Animal number | Sex | Scoring | Animal number | Sex | Scoring |
| 4 5 6 4 5 6 | ♀ ♀ ♀ ♂ ♂ ♂ | 0 0 0 0 0 0 | 7 8 9 7 8 9 | ♀ ♀ ♀ ♂ ♂ ♂ | 4 4 4 4 4 4 | 1 2 3 1 2 3 | ♀ ♀ ♀ ♂ ♂ ♂ | 0 0 0 0 0 0 |
Seven, conclusion (of pressure testing)
Under this experimental condition, powder pin systemic anaphylaxis feminine gender of the present invention, no whole body sensitization.These embodiment only are optimal way of the present invention, do not limit the scope of protection of present invention.
The specific embodiment
Embodiment 1
Chlorogenic acid 0.1g
Baicalin 40g
Sodium chloride 55g
Claim sodium chloride, add an amount of water for injection stirring and make dissolving.Inventory chlorogenic acid, baicalin are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The sodium chloride that adds inventory again, stirring and dissolving adds the injection water to about 550ml.Use 1mol/L hydrochloric acid and 1mol/L sodium hydroxide to regulate the pH value of medicinal liquid, the pH value that makes medicinal liquid is in 6.0～6.8 scope.The active carbon that adds 0.2% amount of liquid medicine is warming up to 70 ℃, stirs 30 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 5.0 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 2
Chlorogenic acid 25g
Baicalin 1.6g
Sodium chloride 67g
Get sodium chloride, add an amount of water for injection stirring and make dissolving.Inventory chlorogenic acid, baicalin are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The sodium chloride that adds inventory again, stirring and dissolving adds the injection water to about 500ml.Use 1mol/L hydrochloric acid and 1mol/L sodium hydroxide to regulate the pH value of medicinal liquid, the pH value that makes medicinal liquid is in 6.0～6.8 scope.The active carbon that adds 1% amount of liquid medicine is warming up to 80 ℃, stirs 40 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 8.0 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 3
Chlorogenic acid 25g
Baicalin 40g
Sodium chloride 35g
Get sodium hydroxide, add an amount of water for injection stirring and make dissolving.Inventory chlorogenic acid, baicalin are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The sodium chloride that adds inventory again, stirring and dissolving adds the injection water to 900ml.Use 1mol/L hydrochloric acid and 1mol/L sodium hydroxide to regulate the pH value of medicinal liquid, the pH value that makes medicinal liquid is in 6.0～6.8 scope.The active carbon that adds 1% amount of liquid medicine is warming up to 75 ℃, stirs 20 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 7.0 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 4
Chlorogenic acid 5.0g
Baicalin 30g
Sodium chloride 63g
Get sodium hydroxide, add an amount of water for injection stirring and make dissolving.Inventory chlorogenic acid, baicalin are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The sodium chloride that adds inventory again, stirring and dissolving adds the injection water to 250ml.Use 1mol/L hydrochloric acid and 1mol/L sodium hydroxide to regulate the pH value of medicinal liquid, the pH value that makes medicinal liquid is in 6.0～6.8 scope.The active carbon that adds 1% amount of liquid medicine is warming up to 75 ℃, stirs 20 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 7.0 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 5
Chlorogenic acid 17.5g
Baicalin 6g
Glucose 78g
It is an amount of to get sodium hydroxide, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The glucose that adds inventory again, stirring and dissolving adds the injection water to about 780ml.The active carbon that adds 1% amount of liquid medicine is warming up to 60 ℃, stirs 30 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 5.5 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 6
Chlorogenic acid 10.0g
Baicalin 23.0g
Glucose 67g
It is an amount of to get sodium hydroxide, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The glucose that adds inventory again, stirring and dissolving adds the injection water to about 1340ml.The active carbon that adds 1% amount of liquid medicine is warming up to 80 ℃, stirs 30 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 6.5 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 7
Chlorogenic acid 14g
Baicalin 17g
Glucose 69g
It is an amount of to get sodium hydroxide, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The glucose that adds inventory again, stirring and dissolving adds the injection water to about 280ml.The active carbon that adds 1% amount of liquid medicine is warming up to 85 ℃, stirs 30 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 7.5 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 8
Chlorogenic acid 5.0g
Baicalin 30.0g
Isochlorogenic acid 8.0g
Wogonoside 4.0g
Glucose 50.5g
It is an amount of to get sodium hydroxide, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin, isochlorogenic acid, wogonoside are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The glucose that adds inventory again, stirring and dissolving adds the injection water to about 1265ml.The active carbon that adds 1% amount of liquid medicine is warming up to 80 ℃, stirs 40 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 8.5 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 9
Chlorogenic acid 17.5g
Baicalin 6g
Isochlorogenic acid 8g
Wogonoside 4g
Sorbitol 60.5g
Weighing sodium hydroxide is an amount of, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin, isochlorogenic acid, wogonoside are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The sorbitol that adds inventory again, stirring and dissolving adds the injection water to about 605ml.The active carbon that adds 1% amount of liquid medicine is warming up to 80 ℃, stirs 50 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 9.0 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 10
Chlorogenic acid 17.5g
Baicalin 30g
Isochlorogenic acid 0.3g
Wogonoside 4.0g
Sorbitol 58.2g
Weighing sodium hydroxide is an amount of, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin, isochlorogenic acid, wogonoside are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The sorbitol that adds inventory again, stirring and dissolving adds the injection water to about 340ml.The active carbon that adds 1% amount of liquid medicine is warming up to 80 ℃, stirs 50 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 6.4 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 11
Chlorogenic acid 17.5g
Baicalin 30.0g
Isochlorogenic acid 8.0g
Wogonoside 0.2g
Sorbitol 44.3g
Weighing sodium hydroxide is an amount of, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin, isochlorogenic acid, wogonoside are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The sorbitol that adds inventory again, stirring and dissolving adds the injection water to about 1100ml.The active carbon that adds 1% amount of liquid medicine is warming up to 80 ℃, stirs 40 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 7.8 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 12
Chlorogenic acid 10g
Baicalin 23g
Isochlorogenic acid 8g
Wogonoside 4g
Sorbitol 45g
Weighing sodium hydroxide is an amount of, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin, isochlorogenic acid, wogonoside are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The sorbitol that adds inventory again, stirring and dissolving adds the injection water to about 180ml.The active carbon that adds 1% amount of liquid medicine is warming up to 50 ℃, stirs 60 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 6.8 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 13
Chlorogenic acid 14g
Baicalin 17g
Isochlorogenic acid 8g
Wogonoside 4g
Mannitol 43g
Weighing sodium hydroxide is an amount of, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin, isochlorogenic acid, wogonoside are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The mannitol that adds inventory again, stirring and dissolving adds the injection water to about 430ml.The active carbon that adds 1% amount of liquid medicine is warming up to 70 ℃, stirs 30 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 7.2 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 14
Chlorogenic acid 14g
Baicalin 23g
Isochlorogenic acid 0.3g
Wogonoside 2g
Mannitol 61g
Weighing sodium hydroxide is an amount of, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin, isochlorogenic acid, wogonoside are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The mannitol that adds inventory again, stirring and dissolving adds the injection water to about 410ml.The active carbon that adds 1% amount of liquid medicine is warming up to 70 ℃, stirs 30 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 7.8 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 15
Chlorogenic acid 14g
Baicalin 23g
Isochlorogenic acid 5g
Wogonoside 0.2g
Mannitol 58g
Weighing sodium hydroxide is an amount of, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin, isochlorogenic acid, wogonoside are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The mannitol that adds inventory again, stirring and dissolving adds the injection water to about 1450ml.The active carbon that adds 1% amount of liquid medicine is warming up to 70 ℃, stirs 30 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 7.8 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 16
Chlorogenic acid 5.0g
Baicalin 30g
Isochlorogenic acid 5g
Wogonoside 2g
Mannitol 55g
Weighing sodium hydroxide is an amount of, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin, isochlorogenic acid, wogonoside are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The mannitol that adds inventory again, stirring and dissolving adds the injection water to about 1300ml.The active carbon that adds 1% amount of liquid medicine is warming up to 80 ℃, stirs 50 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 6.6 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 17
Chlorogenic acid 17.5g
Baicalin 10g
Isochlorogenic acid 5g
Wogonoside 2g
Mannitol 65g
It is an amount of to get sodium hydroxide, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin, isochlorogenic acid, wogonoside are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The mannitol that adds inventory again, stirring and dissolving adds the injection water to about 1300ml.The active carbon that adds 1% amount of liquid medicine is warming up to 80 ℃, stirs 45 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 6.8 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 18
Chlorogenic acid 17.5g
Baicalin 30g
Isochlorogenic acid 0.8g
Wogonoside 2g
Sodium chloride makes finished product to 100g
It is an amount of to get sodium hydroxide, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin, isochlorogenic acid, wogonoside are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The sodium chloride that adds inventory again, stirring and dissolving adds the injection water to about 3000ml.The active carbon that adds 1% amount of liquid medicine is warming up to 70 ℃, stirs 30 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 6.4 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 19
Chlorogenic acid 17.5g
Baicalin 30g
Isochlorogenic acid 5g
Wogonoside 0.4g
Sorbitol makes finished product to 100g
It is an amount of to get sodium hydroxide, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin, isochlorogenic acid, wogonoside are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The sorbitol that adds inventory again, stirring and dissolving adds the injection water to about 3000ml.The active carbon that adds 1% amount of liquid medicine is warming up to 70 ℃, stirs 30 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 6.5 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 20
Chlorogenic acid 10g
Baicalin 23g
Isochlorogenic acid 5g
Wogonoside 2g
Glucose makes finished product to 100g
It is an amount of to get sodium hydroxide, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin, isochlorogenic acid, wogonoside are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.The sorbitol that adds inventory again, stirring and dissolving adds the injection water to about 3000ml.The active carbon that adds 1% amount of liquid medicine is warming up to 70 ℃, stirs 30 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 6.2 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 21
Chlorogenic acid 14g
Baicalin 17g
Isochlorogenic acid 5g
Wogonoside 2g
Sodium hydroxide is an amount of, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin, isochlorogenic acid, wogonoside are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.Add the injection water approximately to 4000ml, doping is an amount of.The active carbon that adds 1% amount of liquid medicine is warming up to 70 ℃, stirs 30 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 6.6 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 22
Chlorogenic acid 14g
Baicalin 23g
Isochlorogenic acid 0.8g
Wogonoside 2g
Sodium hydroxide is an amount of, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin, isochlorogenic acid, wogonoside are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.Add the injection water approximately to 2000ml, add the active carbon of 1% amount of liquid medicine, be warming up to 70 ℃, stirred 30 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 6.6 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Embodiment 23
Chlorogenic acid 14g
Baicalin 23g
Isochlorogenic acid 5g
Wogonoside 0.4g
Sodium hydroxide is an amount of, adds an amount of water for injection stirring and makes dissolving.Inventory chlorogenic acid, baicalin, isochlorogenic acid, wogonoside are dropped in the water for injection, add an amount of 50% sodium hydroxide solution under stirring and make chlorogenic acid and baicalin dissolving, the pH value of control medicinal liquid makes and is lower than 7.5.Add the injection water approximately to 2000ml.The active carbon that adds 1% amount of liquid medicine is warming up to 65 ℃, stirs 45 minutes.Use husky filter stick and vacuum pump with medical filtration.Use the 1mol/L sodium hydroxide to regulate the pH value to 6.2 of medicinal liquid.Use filter with medical filtration, according to the half-finished content of semi-manufactured goods quality standard test, pH value, fill, lyophilization gets the about 100g of lyophilized powder, packing, storage.
Claims (15)
1, a kind of power for intravenous injection, it is characterized in that its main effective ingredient is grouped into by following one-tenth: chlorogenic acid 10-250mg/g, baicalin 16-400mg/g do not contain phillyrin, jasminoidin.
2, powder pin as claimed in claim 1 is characterized in that described active component and content are: chlorogenic acid 50-175mg/g, baicalin 60-300mg/g.
3, powder pin as claimed in claim 2 is characterized in that described active component and content are: chlorogenic acid 100-140mg/g, baicalin 170-230mg/g.
4,, it is characterized in that also comprising the active component of following percentage by weight: isochlorogenic acid 3-80mg/g, wogonoside 2-40mg/g as claim 2 or 3 described powder pins.
5,, it is characterized in that also comprising the active component of following percentage by weight: isochlorogenic acid 8-50mg/g, wogonoside 4-20mg/g as claim 2 or 3 described powder pins.
6, as each described powder pin of claim 1-5, it is characterized in that having added in the pharmaceutical preparation: sodium chloride, glucose, sorbitol or mannitol are as additive.
7, powder pin as claimed in claim 6, the concentration that it is characterized in that additive is between 4%-25%.
8, powder pin as claimed in claim 7, the concentration that it is characterized in that described additive concentration glucose is 5%-10%, other is 10%-15%.
9,, it is characterized in that described pharmaceutical preparation pH value is between 4.0～9.0 as each described powder pin of claim 1-8.
10, as the application of each described powder pin of claim 1-9 in preparation treatment pneumonia medicine.
11, as the application of each described powder pin of claim 1-9 in preparation treatment upper respiratory tract infection medicine.
12, as the application of each described powder pin of claim 1-9 in the preparation antiviral drugs.
13,, it is characterized in that its active ingredient chlorogenic acid measures as follows as each described powder pin of claim 1-9:
Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica;
With ratio is that 10: 90 acetonitrile-0.4% phosphoric acid solution is a mobile phase
The detection wavelength is 327nm
It is an amount of that the preparation precision of reference substance solution takes by weighing the chlorogenic acid reference substance, adds methanol and make the solution that every 1ml contains 0.05mg, shakes up, promptly;
This product is got in the preparation of need testing solution, porphyrize, and it is fixed to get the accurate title of about 0.1g, put in the tool plug conical flask, add 50% methanol 100ml, claim to decide weight, ultrasonic 30min, put cold, weight decided in title again, supplies with 50% methanol to subtract weight loss, shakes up, filter, get the brown measuring bottle of an amount of adding of filtrate and promptly get for test agent solution;
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.
14,, it is characterized in that its effective ingredient baicalin measures as follows as each described powder pin of claim 1-9:
Chromatographic condition and system suitability test octadecyl silane are packing material
With 47: 53: 0.2 methanol-water-phosphoric acid of ratio is mobile phase
Detect wavelength 280nm
It is an amount of that the preparation precision of reference substance solution takes by weighing the baicalin that is dried to constant weight, puts in the 50ml measuring bottle, adds dissolve with methanol, shakes up, and makes the solution that every 1ml contains 60 μ g, promptly;
This product is got in the preparation of need testing solution, porphyrize, and it is fixed to get the accurate title of about 0.1g, put in the 50ml measuring bottle, add 50% methanol 50ml jolting, dissolving, shake up, filter, precision is measured subsequent filtrate 1ml, put in the 25ml measuring bottle, adding 50% methanol to scale, shake up, is the microporous filter membrane of 0.45 μ m with the aperture, filter, promptly;
Accurate respectively reference substance solution and the need testing solution 10 μ l of drawing of algoscopy;
15,, it is characterized in that its preparation technology is according to each described pharmaceutical preparation of claim 1-9:
(1) chlorogenic acid, the preparation technology of isochlorogenic acid: extracting honeysuckle, 4-20 doubly measures 40%-95% ethanol, reflux, extract, 1-5 time, each 0.5-3 hour, filter, merging filtrate, be evaporated to the extractum shape, it is dry to put into vacuum drying oven, and exsiccant Flos Lonicerae extract is dissolved in and is equivalent to its weight 1-20 and doubly measures in the water of volume, aqueous solution extracts 1-5 time with equal amounts of chloroform, chloroform layer discards, and water reuse 0.5-3 doubly measures isoamyl alcohol extraction 1-5 time, merges the isoamyl alcohol layer, concentrating under reduced pressure, vacuum drying gets that to be dissolved in the 40-90% alcoholic solution in right amount an amount of, admixes 200 gram silica gel, 60 ℃ of water-baths volatilize, as the column chromatography for separation sample, take by weighing 200-300 order silica gel 2000 gram, being 40 with ratio: 1-2: 1 chloroform-methanol is a solvent, wet method dress post, sample on the dry method is 40 with ratio: 1-2: 1 chloroform-methanol eluting, and equal-volume is collected, every part of 500ml, gradient elution, every 500ml adds methanol 10ml, and TLC follows the trail of inspection, merge same section, through pillar chromatography and in conjunction with recrystallization technology repeatedly, obtain the chemical compound chlorogenic acid, the chemical compound isochlorogenic acid again;
(2) baicalin, the preparation technology of wogonoside: Radix Scutellariae coarse powder, add 2-10 times of water gaging, reflux, extract, 1-5 time, each 0.5--3 hour, filter, merging filtrate, concentrating under reduced pressure, add 0.5-4mol/LHCl solution and transfer PH=0.5～4.0 in right amount, be incubated 0.5-3 hour, placed 8-24 hour, filter, precipitation adds 4-20 times of water gaging, transfer PH=5.0～10.0, add 0.5-2 times of ethanol again, stir and make dissolving, filter, filtrate is transferred PH=1.0～5.5 with HCl solution, is incubated 0.5-3 hour, places 8-24 hour, filter, precipitation is washed till PH=5.0～10.0 with ethanol, and vacuum drying is got above-mentioned dry thing, mix with 200-300 purpose silica gel, silica gel wet method dress post carries out chromatography, and chloroform-methanol is an eluant, and thin layer is checked eluent, equivalent is collected, every part of 500ml is divided into 2 major parts behind the merging same section: the one, and the chloroform-methanol gradient is 10: 1～90: 1 parts, B is that the chloroform-methanol gradient is 10: 1～1: 1 part, separate out acicular crystal from a part the eluent of 10: 1 sections of chloroform-methanols, further recrystallization obtains chemical compound 2; B partly passes through 200-300 purpose silica gel column chromatography, obtains chemical compound 1 through LH-20 type polydextran gel column chromatography for separation;
(3) preparation technology of said preparation is: the chlorogenic acid of inventory, baicalin etc. are dissolved in the proper amount of water for injection, and making concentration is chlorogenic acid 0.1%-25mg/ml, baicalin 1-40mg/ml, adds isoosmotic adjusting agent, the pH value of control medicinal liquid.Mend to an amount of with water for injection, regulate the pH value of medicinal liquid, the pH value that makes finished product in 4.0～9.0 scope, quality inspection, fill, lyophilization, packing, promptly.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410080023 CN1257723C (en) | 2004-09-24 | 2004-09-24 | Powder injection for intravenous injection and its preparing method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410080023 CN1257723C (en) | 2004-09-24 | 2004-09-24 | Powder injection for intravenous injection and its preparing method |
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| CN1616051A CN1616051A (en) | 2005-05-18 |
| CN1257723C true CN1257723C (en) | 2006-05-31 |
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| CN 200410080023 Expired - Fee Related CN1257723C (en) | 2004-09-24 | 2004-09-24 | Powder injection for intravenous injection and its preparing method |
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