CN1883523A - A compound lyophilized powder injection and preparation method thereof - Google Patents

Abstract

Disclosed is a freeze dried powder injection and its preparation process, which is prepared from the following raw materials (by weight ratio): Picrasma quassioides 1-10, green chiretta 1-10. The preparing process comprises the steps of alcohol extracting, eluting, drying, and freeze drying.

Description

A kind of compound lyophilized powder injection and preparation method thereof

(1) technical field: a kind of compound lyophilized powder injection and preparation method thereof, belong to Chinese medicine preparation, specifically be to be the medicament that raw material is made with the botanical herbs, and its preparation method.

(2) background technology: show that according to the relevant statistics of health ministry the prevalence of China's respiratory system disease is about about 6.94%, promptly the whole nation has more than 8,000 ten thousand people to suffer from respiratory system disease every year.On disease constituted, upper respiratory tract infection accounted for the major part of whole respiratory system disease.And from the age, an old Shaozheng is the age bracket occurred frequently of respiratory system disease.Acute upper respiratory tract infection easily causes the hyperpyrexia emergency case, and more than 39 ℃, whole body is scorching hot, excessive thirst, rapid pulse are that hyperpyrexia convulsion takes place easily for cardinal symptom, especially child, and life is caused serious threat with body temperature.Acute upper respiratory tract infection, hyperpyrexia class disease are except the medication of suiting the medicine to the illness, and the infection medication is essential, especially based on antibiotic.But enhancing along with bacterial drug resistance, the toxic and side effects of Western medicine is outstanding all the more, and the Western medicine antibiotic is at a loss what to do substantially to this type of disease that unknown cause causes, antibiotic is to have specificly for the kind of antibacterial or virus, and the diseases such as heating that some unknown causes are produced do not have specific treatment way substantially.It is rapid to infect class disease frequently-occurring disease, cause complication and critical illness easily, severe form and antibiotic limitation under the pressure of this type of disease, the exploitation natural drug, with the multicomponent of natural drug, the therapeutic modality of many target spots, slow down the generation of Resistant strain, improve this type of treatment of diseases effect and seem very urgent.Adopt diseases such as treatment by Chinese herbs upper respiratory tract infection, hyperpyrexia, good effect, toxic and side effects is little, have no drug resistance, and need not hypersensitive test, this is a big speciality of the traditional Chinese medical science, all is to refine to be processed into from China's Chinese medicine as antipyretic-detoxicant preparations such as the QINGKAILING of development newly developed in recent years, SHUANGHUANLIAN, Herba Houttuyniae, Andrographolides, characteristics such as it is steady to have therapeutical effect, and side effect is little.And Chinese medicine is with regard to the treatment infectious disease, looses, conventional dosage forms Yin Qihei such as pellet, cream, big, thick profile and produce effects speed are slow, use inconvenience, and the unsettled shortcoming of curative effect can not satisfy clinical and social demand.It is the first-selected route of administration of treatment hyperpyrexia emergency case that the injection injection is brought down a fever, especially intravenous injection or instillation, its bioavailability 100%, produce effects is fast, and injection is more suitable for clinical needs, but injection is because of the unstability of its preparation, clinical practice is limited to, and make troubles for storage and transportation, fragment and microgranule pollute medicinal liquid easily when cutting saw injection vial in addition, damage to the patient.Therefore, be badly in need of efficient, quick-acting, safe high-grade Chinese medicine preparation on the market and fill up this blank.

(3) summary of the invention: the problem to be solved in the present invention be exactly provide a kind of at above deficiency and treat that curative effect of disease such as upper respiratory tract infection, hyperpyrexia are good, produce effects soon, safely, the Chinese medicine preparation that has no drug resistance, and its preparation method.Its technical scheme is as follows:

Take by weighing each crude drug by following weight portion: Ramulus Et Folium Picrasmae 1～10, Herba Andrographis 1～10.

Preparation method may further comprise the steps:

1) get Ramulus Et Folium Picrasmae and be ground into coarse powder, add 6～8 times of amount 75%--80% ethanol extractions 2～3 times with Folium Andrographis, each 1～2 hour, merge extractive liquid, filtered filtrate recycling ethanol to 1: 1.5;

2) in above-mentioned filtrate, add the active carbon reflux 25～30 minutes of inventory 5～6%, filter while hot, and use the washing with alcohol filtering residue, filtration, merging filtrate;

3) add the ethanol of 3～4 times of inventorys in above-mentioned filtrate, stir, left standstill 24～36 hours, filter, filtrate recycling ethanol is made fluid extract;

4) in fluid extract, add the Diluted Alcohol dissolving, filter, get filtrate by macroporous adsorbent resin, it is colourless that water is eluted to effluent, the reuse ethanol elution, collect eluent, reclaim ethanol, concentrate drying under reduced pressure to doing, add the water for injection dissolving fully, add the pharmaceutic adjuvant lactose, Dextran 40, aminoacid, sodium chloride, mannitol one or two or more kinds, consumption is the 20-200% of extract amount, fully stirs, regulate its pH value to 5-7, the active carbon of adding 0.5%～0.6% is incubated 15～20 minutes down at 50-60 ℃, filters, packing, lyophilization, gland, promptly.

Perhaps take by weighing each crude drug by following weight portion:

Ramulus et Folium Picrasmae 20g～200g, Herba Andrographis extract 20g～200g.

Preparation method is to get Ramulus Et Folium Picrasmae and Herba Andrographis extract, adds the water for injection dissolving fully, adds pharmaceutic adjuvant, fully stirs, regulate its pH value to 5-7, add 0.5%～0.6% active carbon, be incubated 15～20 minutes down, filter at 50-60 ℃, packing, lyophilization, gland, promptly.

Compared with prior art the beneficial effect that has of the present invention is:

1, product of the present invention is highly purified natural medicinal formulations, and Ramulus Et Folium Picrasmae nature and flavor bitter cold is slightly poisonous, return lung, large intestine channel, function are anti-inflammation with curing mainly, the damp eliminating detoxifcation, the Herba Andrographis bitter in the mouth, cold in nature, GUIXIN, lung, large intestine, urinary bladder channel has heat-clearing and toxic substances removing, the effect of removing heat from blood detumescence, the two collaborative multicomponent with natural drug, the therapeutic modality of many target spots slows down the generation of Resistant strain, and therapeutic effect is good, instant effect, toxic and side effects is little, and especially gastrointestinal reaction is little, drug safety, can better treat the heating repeatedly that upper respiratory tract infection etc. causes, persistent fever class disease, solve the Western medicine antibiotic because of agnogenio really can't symptomatic treatment and a difficult problem such as side effect is bigger.

2, this product is advanced lyophilized injectable powder, compares with injection, has following advantage:

1) the products obtained therefrom quality is loose, can dissolve rapidly after adding water, and the said preparation water content is low, and powder pin packing can keep vacuum or filling noble gas, helps strengthening stability of drug;

2) its sterilization process is that filtering with microporous membrane by the different apertures of multilamellar obtains can be used for after the packing the directly sterile liquid medicine of lyophilizing embedding, and avoided the High Temperature High Pressure of injection sterilization, thereby give to thermo-responsive or in aqueous solution unsettled medicine a good dosage form is provided;

3) can be mixed with the solution of variable concentrations because of the difference of clinical application, satisfy the needs of different way of administration;

4) packaging volume is little, and packaging material require low, have reduced packing cost and cost of transportation;

Fragment and particle contamination medicinal liquid guarantee patient's drug safety when 5) having avoided cutting saw injection vial.

Medicament selection Ramulus Et Folium Picrasmae of the present invention and Herba Andrographis make up, and their combination makes its effect produce synergism, thereby can effectively treat upper respiratory tract infection, hyperpyrexia class disease:

Ramulus Et Folium Picrasmae [Picrasmaquassiodes (D.Don) Ben] is a Simarubaceae Quassia plant.Widely distributed in China, aboundresources.The nature and flavor bitter cold of this medicine, slightly poisonous.Return lung, large intestine channel.Function is anti-inflammation with curing mainly, the damp eliminating detoxifcation.Its pharmacological research and clinical practice are as follows: 1, antibiotic: the Ramulus Et Folium Picrasmae alkaloid has antifungic action, and external antifungal experiment finds that Rhomotoxine has bacteriostasis to hemolytic group B streptococcus, staphylococcus aureus, sonne bacillus, bacillus subtilis and sarcina; 2, antiinflammatory: mice ear inflammation due to the Ramulus Et Folium Picrasmae preparation xylol and Ovum Gallus domesticus album foot are swollen to have obvious inhibition to use; 3, analgesic: rabbit intramuscular injection Ramulus Et Folium Picrasmae preparation is to typhoid fever, and heating has refrigeration function due to the paratyphoid fever mixed vaccine; 4, toxicity: Rhomotoxine is not seen obvious influence to rat growthing development, liver, renal function, hemogram and parenchymatous organ, the heart, liver, lung, kidney.

Herba Andrographis [Andrographispaniculata (Burm.f) Nees] has another name called Herba Andrographitis, Herba vallisneriae Spiralis, and longleaf rabdosia stem, India's grass etc. are Acanthaceae Herba Andrographis platymiscium.Main component has lactone, sterol, multiple flavone compound, alkaloid etc.Bitter in the mouth, cold in nature, GUIXIN, lung, large intestine, urinary bladder channel has heat-clearing and toxic substances removing, and the effect of removing heat from blood detumescence is that a kind of source is wide, the Chinese herbal medicine commonly used that toxic and side effects is few.Its pharmacological research and clinical practice are as follows: 1, antibiotic, antivirus action: the Herba Andrographis water decoction has certain inhibitory action to bacillary dysentery, leptospira, streptococcus pneumoniae etc.Bluff the Chuanhuning injection that amber acid half ester (DAS) monopotassium salt makes by dehydrorographolide and staphylococcus aureus and Staphylococcus albus are had inhibitory action with agar plate method, the intravenous injection of andrographolide iodate injection is to the lagophthalmos pathological changes of streptococcus pneumoniae local infection, can significantly shorten the course of disease, quicken the absorption of inflammation.2, antiinflammatory action: the effective ingredient of Herba Andrographis, Herba Andrographis first, second, third, fourth element all have antiinflammatory action in various degree, can suppress the hyperfunction and exudation of the early stage capillary permeability of acute inflammation; 3, refrigeration function: Herba Andrographis and interior vinegar constituents thereof are for the rabbit or 2 that generates heat due to typhoid fever, the paratyphoid vaccine, the rat that generates heat due to the 2, 4-dinitrophenol has certain refrigeration function, can postpone the body temperature rise time to fever in rabbit due to while pneumonia infection diplococcus and the Hemolytic streptococcus culture, weaken the body temperature rise degree; 4, andrographolide and derivant thereof do not have overt toxicity in vivo.Sub-acute toxicity test shows that andrographolide 1g/kg gavages, and continuous 7 days of every day 1 time, rat or rabbit is not had overt toxicity; Chuanhuning injection 84mg/kg lumbar injection every day 1 time, continuous 10 days to the rat free of toxic effects; 5, be usually used in treating multiple infectious disease such as flu, tonsillitis, bronchitis, acute bacillary dysentery, gastroenteritis clinically.

For proving beneficial effect of the present invention and practicality, the inventor has made following pharmacodynamics and toxicity research to product of the present invention:

Method: observe the influence of product of the present invention to streptococcus pneumoniae, streptococcus, dysentery bacterium, staphylococcus aureus, escherichia coli, salmonella; After observing drug administration by injection, to the influence of LPS pyrogenicity rabbit body temperature,, carrageenin is caused the influence of rat paw edema to the inductive exothermic reaction of beer yeast, xylol causes the influence of mice ear, the influence that the mouse peritoneal capillary permeability of Dichlorodiphenyl Acetate mediation increases; The acute toxic reaction of mice behind the observation drug administration by injection.

Strain and animal: strain: dysentery bacterium (clinical samples isolate group is provided by the Clinical Laboratory center); Streptococcus pneumoniae, streptococcus, Escherichia coli, staphylococcus aureus, salmonella are from Nat'l Pharmaceutical ﹠ Biological Products Control Institute.Bacterium liquid: each reference culture is inoculated in the 10ml nutrient broth medium with inoculating loop, cultivates 18～24h for 37 ℃, promptly.Culture medium: nutrient broth medium, nutrient agar.Animal: healthy closed colony new zealand rabbit, male, body weight 2.0～2.6kg; Kunming mouse, body weight 18～22g, male and female dual-purpose; The Wistar healthy rat, body weight 180～220g, male and female dual-purpose; Provide (production licence numbering: 015) by Sichuan Province's Experimental Animal Center.

The mensuration of minimum inhibitory concentration (MIC) adopts double dilution method, getting an amount of product freeze-dried powder solution of the present invention adds and makes in the nutrient broth medium at double than 5 of dilution factors i.e. (10,5,2.5,1.25,0.625mg/ml), more than each dilution factor respectively draw 1.0ml and add in 7 blank test tubes of sterilization, altogether 7 groups 5 dilution dosing meat soup solution; In every group of dosing meat soup test tube, add streptococcus pneumoniae, streptococcus, dysentery bacterium, staphylococcus aureus, escherichia coli, various each 0.1ml of test organisms liquid of salmonella respectively, cultivate 18～24h (result such as table 1) for 37 ℃.

Table 1 double dilution method is measured anti-inflammatory and choleretic solution MIC experimental result

Tried bacterium	Drug quality concentration (mg/ml)
						10	5	2.5	1.25	0.625
	Streptococcus pneumoniae streptococcus dysentery bacterium staphylococcus salmonella escherichia coli	- - - - - -	- - - - - -	- - - - - -	- - - - - -						- - - - + +

Annotate: "+" expression test pipe range bacterium; The not long bacterium of "-" expression developmental tube

Product freeze-dried powder solution of the present invention as can be seen from Table 1 is 0.625mg/ml to the minimum inhibitory concentration MIC of streptococcus pneumoniae, streptococcus, staphylococcus aureus, dysentery bacterium, is 1.25mg/ml to the minimum inhibitory concentration MIC of escherichia coli, salmonella.

Bacteriostatic test:

Adopt the agar plate diffusion method.With 10 on diameter 90cm culture dish, 2 every group, add the nutrient agar sterilized and melted, each 15ml of every ware, after waiting to solidify, every group of ware 5ml that tiles contains the Nutrient agar of every kind of bacterium; 5 Oxford cups of each dull and stereotyped equidistant placement, the product freeze-dried powder solution of the present invention that splashes into 5 concentration in the table 3 are cultivated 16～18h for 37 ℃ in the cup of Oxford, measure inhibition zone diameter, calculating mean value (the results are shown in Table 2).

Table 2 agar plate diffusion method result of the test

Tried bacterium	Tried the diameter (D/mm) of inhibition zone under each concentration of bacterium (mg/ml)
						10	8	6	5	4
	Streptococcus pneumoniae streptococcus dysentery bacterium staphylococcus salmonella escherichia coli	25.36 23.77 24.12 16.79 15.87 15.01	23.09 21.88 22.13 15.66 15.08 14.33	22.59 21.12 21.78 14.57 14.56 13.25	21.63 19.42 19.99 13.38 13.77 12.89						19.55 17.98 18.67 12.79 12.86 12.23

By table 2 as seen, product freeze-dried powder solution of the present invention is the strongest to the bacteriostasis of streptococcus pneumoniae, streptococcus, dysentery bacterium, and the bacteriostasis of salmonella, staphylococcus aureus is taken second place, and is the most weak to colibacillary bacteriostasis.

Product freeze-dried powder of the present invention is to the influence of LPS pyrogenicity rabbit body temperature:

Select the qualified rabbit of body temperature (38.5～39.5 ℃ of normal body temperatures) before the experiment for use, grouping and dosage see Table 3.Experiment is worked as day interval 1h continuous measurement rabbit body temperature 2 times, gets its meansigma methods as basal body temperature.Every subsequently rabbit injection designs the medicine of dosage, 10min auricular vein injection endotoxin 0.2ug/kg (being diluted to 0.2mg/L) behind the medicine with the apyrogeneity normal saline, respectively at after the administration 30,60,120, each take temperature of 180min 1 time, calculate body temperature and basal body temperature difference (the results are shown in Table 3) behind the medicine.

The influence of table 3 pair LPS pyrogenicity rabbit body temperature (x ± s, ℃)

Group	Quantity (only)	Dosage mg/kg	Basal body temperature	Different time body temperature difference after the administration
								30min	60min	120min	180min
				Dosage Chinese medicine low dosage in the model group aspirin group Chinese medicine high dose Chinese medicine	7 7 7 7 7	- 200 10 5 2.5	39.11±1.02 39.23±0.96 38.98±1.09 38.87±1.21 39.19±1.31					0.42±0.12 0.25±0.08* 0.13±0.09** 0.24±0.07* 0.26±0.13*	0.95±0.24 0.35±0.13** 0.27±0.09** 0.37±0.15** 0.42±0.17**	1.55±0.31 0.52±0.18** 0.42±0.21** 0.57±0.15** 0.72±0.26*	1.46±0.27 0.75±0.24* 0.72±0.26* 0.85±0.29* 0.93±0.31*

Annotate: experiment back administration treated animal and model group animal in corresponding index are relatively learned by statistics and handle (Dunnett t check), and there is statistical significance * P＜0.05, * * P＜0.01.

Table 3 result shows, high, medium and low dosage group of product freeze-dried powder of the present invention and positive control drug aspirin group all can significantly be resisted the exothermic reaction of endotaxin induction, with model control group significant difference (P＜0.05 or P＜0.01) is arranged relatively, and product freeze-dried powder of the present invention onset time of high dose is early than aspirin.

Product freeze-dried powder of the present invention heats the influence of rat temperature to beer yeast:

The qualified SD rat (normal body temperature 36.4-38.2 ℃) of screening body temperature is male before the test, and grouping and dosage see Table 4.Test is got its meansigma methods as basal body temperature when 1 hour continuous measurement rat temperature of day interval 2 times.Every subsequently Mus nape portion subcutaneous injection 100 grams per liter beer yeast normal saline suspensions 1 gram/kilogram was measured after the modeling and is heated body temperature in 6 hours, was used for test with the intensification value greater than 1 ℃ of rat.The medicine of dosage is given and designed to every rat, measured 1 body temperature every 1 hour behind the medicine, and continuous 4 times, body temperature and basal body temperature difference after the calculating administration.

Table 4 pair beer yeast heats the influence of rat temperature

Group	Quantity (only)	Dosage mg/kg	Basal body temperature	Different time body temperature difference after the administration
								60min	120min	180min	240min
				Dosage Chinese medicine low dosage in the model group aspirin group Chinese medicine high dose Chinese medicine	10 10 10 10 10	- 150 8 4 2	37.11±0.52 37.23±0.46 37.38±0.59 37.17±0.61 37.19±0.49					1.92±0.32 0.85±0.38** 0.79±0.35** 0.89±0.39** 1.05±0.41*	1.75±0.29 0.75±0.33** 0.69±0.31** 0.78±0.35** 0.99±0.39*	1.65±0.33 0.52±0.28** 0.51±0.27** 0.59±0.30** 0.82±0.32*	1.56±0.28 0.65±0.24** 0.49±0.22** 0.55±0.26** 0.75±0.27*

Annotate: experiment back administration treated animal and model group animal in corresponding index are relatively learned by statistics and handle (Dunnett t check), and there is statistical significance * P＜0.05, * * P＜0.01.

Fruit shows table 4, high, medium and low dosage group of product freeze-dried powder of the present invention and positive control drug aspirin group all can significantly be resisted the inductive exothermic reaction of beer yeast, with model control group significant difference (P＜0.05 or P＜0.01) is arranged relatively, and prolong the onset time of Chinese drug-treated group than aspirin.

Product freeze-dried powder of the present invention causes the influence of rat paw edema to carrageenin:

Select the Wistar rat for use, grouping and dosage see Table 5.Each treated animal drug administration by injection every day 1 time, continuous 3d, before the last administration, volumetric method is measured the right back toes normal volume of rat (is sign with the ankle joint), 15min after the last administration, right back toes subcutaneous injection 10g/L carrageenin suspension 0.08ml/ only, respectively at after the administration 1,2,4,6h measures right back toes volume 1 time, calculate and cause scorching back different time toes swelling degree differences (the results are shown in Table 5).

Table 5 pair carrageenin causes the influence (x ± s) of rat paw edema

Group	Quantity (only)	Dosage mg/kg	Cause scorching preceding toes volume	Different time foot swelling difference after the administration
								1h	2h	4h	6h
				Dosage Chinese medicine low dosage in the model group aspirin group Chinese medicine high dose Chinese medicine	10 10 10 10 10	- 100 4 2 1	1.45±0.11 1.45±0.11 1.45±0.11 1.45±0.11 1.45±0.11					0.25±0.07 0.15±0.03* 0.11±0.04** 0.14±0.03* 0.17±0.05*	0.56±0.11 0.35±0.12* 0.27±0.11** 0.33±0.12* 0.39±0.13*	0.94±0.15 0.70±0.13* 0.62±0.12* 0.71±0.14* 0.80±0.15	0.96±0.17 0.68±0.16* 0.64±0.14* 0.70±0.15* 0.82±0.19

Annotate: experiment back administration treated animal and model group animal in corresponding index are relatively learned by statistics and handle (Dunnett t check), and there is statistical significance * P＜0.05, * * P＜0.01.

Table 5 result shows that the high, medium and low dosage group of product freeze-dried powder of the present invention has significant inhibition foot swelling effect in causing scorching back 4h, similar to the effect of aspirin group (comparing P＜0.05 or P＜0.01 with model control group).

Product freeze-dried powder xylol of the present invention causes the influence of mice ear:

Select Kunming mouse for use, male and female half and half, grouping and dosage see Table 6.Each organizes continuous drug administration by injection 3d, 15min after the last administration, the inside and outside two sides of every Mus auris dextra is evenly smeared each 15uL of dimethylbenzene and is caused inflammation, left side ear contrast does not deal with, take off cervical vertebra execution mice after causing scorching 3h, cut ears, get the left-right symmetric auricle with diameter 8.0mm card punch, electronic balance claims quality, and left and right sides ear weight difference is counted swelling degree (the results are shown in Table 6).

Table 6 xylol causes the influence (x ± s) of mice ear

Group	Number of animals (only)	Dosage mg/kg	Left and right sides auricle weight difference mg
				Dosage Chinese medicine low dosage in the model group aspirin group Chinese medicine high dose Chinese medicine	12 12 12 12 12	- 150 6 3 1.5	8.12±1.34 2.33±0.65** 2.54±0.48** 3.55±1.06** 4.33±1.37*

Annotate: experiment back administration treated animal and model group animal in corresponding index are relatively learned by statistics and handle (Dunnett t check), and there is statistical significance * P＜0.05, * * P＜0.01.

Table 6 result shows that 3 dosage of product freeze-dried powder of the present invention all can significantly suppress the mice ear (comparing P＜0.01 with model control group) due to the dimethylbenzene, point out product freeze-dried powder of the present invention that the acute inflammation of inhibition transudation is arranged.

The influence that the mouse peritoneal capillary permeability of product freeze-dried powder Dichlorodiphenyl Acetate mediation of the present invention increases:

Select Kunming mouse for use, male and female half and half, grouping and dosage see Table 7.Each organizes continuous drug administration by injection 3d, behind the last administration 15min, tail vein injection 5g/L azovan blue 50mg/kg, lumbar injection acetic acid 20ml/kg takes off cervical vertebra and puts to death mice behind the 30min immediately, every Mus intraperitoneal injection of saline 5ml, open the abdominal cavity and collect 3mL abdominal cavity washing liquid, be settled to 10ml, the centrifugal 10min of 3000r/min, collect supernatant, UV-260 type ultraviolet divides spectrophotometer to measure D value (590nm) (the results are shown in Table 7).

The influence that the mouse peritoneal capillary permeability of table 7 Dichlorodiphenyl Acetate mediation increases (x ± s)

Group	Number of animals (only)	Dosage mg/kg	Vascular permeability
				Dosage Chinese medicine low dosage in the model group aspirin group Chinese medicine high dose Chinese medicine	12 12 12 12 12	- 150 6 3 1.5	0.142±0.025 0.091±0.014** 0.089±0.015** 0.094±0.017** 0.101±0.024*

Annotate: experiment back administration treated animal and model group animal in corresponding index are relatively learned by statistics and handle (Dunnett t check), and there is statistical significance * P＜0.05, * * P＜0.01.

Table 7 result shows that the mouse peritoneal capillary permeability that 3 dosage of product freeze-dried powder of the present invention all can significantly suppress the acetic acid mediation increases the effect of pointing out this medicine to have anti-acute inflammation to ooze out.

Acute toxicity test in mice:

After the injected in mice product freeze-dried powder of the present invention medicinal liquid 300mg/kg body weight, in 7 days, grow health, fleshiness, dense and glossy by hair, be close to its body, bright and flexible, the N/R secretions of eyes, the crissum cleaning, it is normal to ingest, the extremity stalwartness, spontaneous activity is normal, and body weight increases gradually, compares no significant difference with the blank group.Do not see animal dead and significant toxicity in 7 days.To dissect behind the sacrifice of animal when experiment finishes, observe vitals such as the heart, liver, spleen, lung, kidney, gastrointestinal and there is no obvious pathological change (body weight change sees table 8 for details).

The acute toxicity test of table 8 product freeze-dried powder of the present invention to the influence of mice body weight (x ± s, g)

Group	Number of animals (only)	Before the experiment	After the experiment
				The female matched group of the male administration group of the male matched group of administration group is female	8 8 8 8	20.2±0.9 19.9±0.8 19.7±0.7 19.8±0.8	27.1±2.4 28.3±2.8 24.7±2.2 25.6±1.9

Annotate: the body weight of administration treated animal after experiment, with control animals at the weight ratio of corresponding time, learn by statistics and handle (Dunnett t check), its male and female there are no significant difference, P is equal＞0.05.

From last table as seen, the body weight after product freeze-dried powder administration group experiment of the present invention finishes is a little less than matched group, but not statistically significant, may be because the 1st day heavy dose of drug administration by injection has certain influence to mice, its speed of growth has slowed down.After the mice maximum dosage-feeding injection product freeze-dried powder medicinal liquid 300mg/kg body weight of the present invention, animal does not occur dead and significantly toxic reaction in 7 days.Results suggest, the acute toxicity of product freeze-dried powder of the present invention is less, application safety.

The result shows: product freeze-dried powder of the present invention is in various degree antibacterial activity in vitro to antibacterial not of the same race, and is the strongest to bacteriostasis and the bactericidal action of streptococcus pneumoniae, streptococcus and dysentery bacterium, along with its bacteriostasis of increase of drug level also strengthens; The high, medium and low dosage group of product freeze-dried powder of the present invention all can significantly be resisted the exothermic reaction of endotaxin induction, all can significantly resist the inductive exothermic reaction of beer yeast, all can in causing scorching back 4h, significant inhibition foot swelling effect be arranged, all can significantly suppress the mice ear due to the dimethylbenzene, the mouse peritoneal capillary permeability that all can significantly suppress the acetic acid mediation increases; After the injected in mice compound quassia freeze-dried powder medicinal liquid 300mg/kg body weight, animal does not occur dead and significantly toxic reaction in 7 days.

Conclusion:

This Preliminary experiment results shows that product freeze-dried powder of the present invention has good antibiotic, antibacterial, analgesic, antiinflammatory action; Anxious poison experiment shows that the acute toxicity of product freeze-dried powder of the present invention is less, application safety.

(4) description of drawings:

Accompanying drawing is a process chart of the present invention.

(5) specific embodiment:

Embodiment one:

Take by weighing each crude drug by following weight portion: Ramulus Et Folium Picrasmae 8, Herba Andrographis 5

Preparation method may further comprise the steps:

1) get Ramulus Et Folium Picrasmae and be ground into coarse powder, add 7 times of amount 80% ethanol extractions 3 times with Folium Andrographis, each 2 hours, merge extractive liquid, filtered filtrate recycling ethanol to 1: 1.5;

2) in above-mentioned filtrate, add the active carbon reflux 30 minutes of inventory 5～6%, filter while hot, and use the washing with alcohol filtering residue, filtration, merging filtrate;

3) ethanol of 3 times of inventorys of adding in above-mentioned filtrate stirs, and leaves standstill 24 hours, filters, and filtrate recycling ethanol is made fluid extract;

4) in fluid extract, add the Diluted Alcohol dissolving fully, filter, get filtrate and pass through macroporous adsorbent resin, it is colourless that water is eluted to effluent, and the reuse ethanol elution is collected eluent, reclaim ethanol, concentrate drying under reduced pressure, add the water for injection dissolving fully to doing, add lactose 100g, fully stir, regulate its pH value to 5-7, the active carbon that adds inventory 0.5%～0.6% is incubated 15 minutes down at 50-60 ℃, filters packing, lyophilization, gland, promptly.Above-mentioned inventory is meant the weight of Ramulus Et Folium Picrasmae and Herba Andrographis.

Embodiment two:

Take by weighing each crude drug by following weight: Ramulus et Folium Picrasmae 100g, Herba Andrographis extract 80g.Preparation method is to add the water for injection dissolving fully in Ramulus et Folium Picrasmae and Herba Andrographis extract, adds 100g mannitol, fully stirs, and regulates its pH value to 5-7, add the 1g active carbon, be incubated 15～20 minutes down, filter packing at 50-60 ℃, lyophilization, gland, promptly.

Product of the present invention also can be prepared into injection as required.

Claims (5)

1, a kind of compound lyophilized powder injection is characterized in that it mainly is by comprising that following raw medicaments in portion by weight makes: Ramulus Et Folium Picrasmae 1～10, Herba Andrographis 1～10.

2, a kind of preparation method of compound lyophilized powder injection is characterized in that may further comprise the steps:

1) get Ramulus Et Folium Picrasmae and be ground into coarse powder, add 6～8 times of amount 75%--80% ethanol extractions 2～3 times with Folium Andrographis, each 1～2 hour, merge extractive liquid, filtered filtrate recycling ethanol to 1: 1.5;

2) in above-mentioned filtrate, add the active carbon reflux 25～30 minutes of inventory 5～6%, filter while hot, and use the washing with alcohol filtering residue, filtration, merging filtrate;

3) add the ethanol of 3～4 times of inventorys in above-mentioned filtrate, stir, left standstill 24～36 hours, filter, filtrate recycling ethanol is made fluid extract;

4) add the Diluted Alcohol dissolving fully in fluid extract, filter, get filtrate by macroporous adsorbent resin, it is colourless that water is eluted to effluent, the reuse ethanol elution is collected eluent, reclaims ethanol, concentrate drying under reduced pressure to doing, add the water for injection dissolving fully, add pharmaceutic adjuvant, fully stir, regulate its pH value, add 0.5%～0.6% active carbon to 5-7, be incubated 15～20 minutes down at 50-60 ℃, filter packing, lyophilization, gland, promptly.

3, the preparation method of compound lyophilized powder injection according to claim 2 is characterized in that may further comprise the steps:

1) get Ramulus Et Folium Picrasmae and be ground into coarse powder, add 7 times of amount 80% ethanol extractions 3 times with Folium Andrographis, each 2 hours, merge extractive liquid, filtered filtrate recycling ethanol to 1: 1.5;

2) in above-mentioned filtrate, add the active carbon reflux 30 minutes of inventory 5～6%, filter while hot, and use the washing with alcohol filtering residue, filtration, merging filtrate;

3) ethanol of 3 times of inventorys of adding in above-mentioned filtrate stirs, and leaves standstill 24 hours, filters, and filtrate recycling ethanol is made fluid extract;

4) add the Diluted Alcohol dissolving in fluid extract, filter, get filtrate by macroporous adsorbent resin, it is colourless that water is eluted to effluent, the reuse ethanol elution is collected eluent, reclaims ethanol, be dried to driedly, add the water for injection dissolving fully, add pharmaceutic adjuvant, fully stir, regulate its pH value, add 0.5%～0.6% active carbon to 5-7, be incubated 15 minutes down at 50-60 ℃, filter packing, lyophilization, gland, promptly.

4, a kind of compound lyophilized powder injection is characterized in that it mainly is by comprising that following raw medicaments in portion by weight makes: Ramulus et Folium Picrasmae 20g～200g, Herba Andrographis extract 20g～200g.

5, a kind of preparation method of compound lyophilized powder injection is characterized in that getting Ramulus Et Folium Picrasmae and Herba Andrographis extract, adds the water for injection dissolving fully, add pharmaceutic adjuvant, fully stir, regulate its pH value, add 0.5%～0.6% active carbon to 5-7, be incubated 15～20 minutes down at 50-60 ℃, filter packing, lyophilization, gland, promptly.