CN1861087B - High purity wild Radix scutellariae glucoside medicine composition, and application of making medicine to treat diseases of cardiovascular and cerebrovascular - Google Patents

Abstract

A composite medicine in the form of solution for treating cardiovascular and cerebrovascular diseases features that it contains high-purity wild-scutelloside and pH regulator proportionally and the content of said high-purity wild-scutelloside can be stabilized for at least 18 months.

Description

A kind of high-purity wild baicalin pharmaceutical composition and the medical usage aspect the preparation heart, cerebrovascular drug thereof

Technical field

The present invention relates to a kind of high-purity wild baicalin pharmaceutical composition and the medical usage aspect the preparation heart, cerebrovascular drug thereof.

Background technology

Scutellarin (Scutellarin), have another name called scutellarin, scutellarin, scutellarin, lamp-dish flower acetic, seal baicalin and (come from the active ingredient of autonomic drug handbook: People's Health Publisher's in March, 1986 front page, p946), chemistry by name 4 ', 5,6-trihydroxyflavone-7-O-glucuronide.Its structural formula is as follows:

At the high Radix Scutellariae Scutellaria of lip plant altissima L. leaf, Radix Scutellariae S.baicalensisGeorgi stem and leaf, scutellarin all has certain content in the plants such as short booth Herba Erigerontis aceris (Herba Erigerontis, Herba Erigerontis) Erigeronbreviscapus (Vant.) Hand-Mazz of feverfew.

The Herba Erigerontis prime system is isolated flavonoids effective constituent from Herba Erigerontis Erigeron breviscapus (Vant.) Hand-Mazz herb, is a kind of flavonoid effective part group, wherein mainly contains lamp-dish flower acetic; The breviscapine of buying from the market at present is the mixture of scutellarin and breviscapine, analyzes through HPLC, and lamp-dish flower acetic content is about 90%, and majority is lower than 90%.

In ministry standard, what the content assaying method of breviscapine adopted is ultraviolet spectrophotometry (abbreviation ultraviolet method), measures trap at 335nm wavelength place, is 570 to calculate that to contain lamp-dish flower acetic be 95%-105% by the absorptance of lamp-dish flower acetic.

In recent years, in the research of high-efficient liquid phase technique mensuration breviscapine, find, the commercially available breviscapine crude drug (ultraviolet method is measured more than 95%) that meets the national ministry standard that is always adopting, main component lamp-dish flower acetic (having another name called scutellarin) content employing HPLC method wherein are determined in the very big scope fluctuate (the commercially available breviscapine HPLC method mensuration scutellarin actual content in the legal source of a plurality of manufacturing enterprises production that buys in studying according to us is 50%-80%).

The deutero-various preparations of breviscapine have been widely used in the treatment cardiovascular and cerebrovascular disease clinically, evident in efficacy, but its injection poor stability, period of storage is short, there are phenomenons such as caloric response, skin pruritus, erythra to take place when individual patient is with the back clinically, the appearance of these phenomenons does not eliminate in process of production mainly due to crude drug due to the some of them impurity, and this problem fails over more than 30 year to solve since the Herba Erigerontis product comes out always.

Recently, Chinese patent CN1640409A discloses a kind of highly purified breviscapine B injection liquid, and its principal character is to contain the injection supplementary material composition that lamp-dish flower acetic or its physiologically acceptable salt and pharmacology allow.Wherein said adjuvant is any one or more a mixture of aminoacid, glucose, lactose, sucrose, trehalose, fructose, maltose, xylitol, sterin, mannitol, dextran, sodium chloride, calcium gluconate or phosphate, phospholipid or derivatives thereof, cyclodextrin and derivant thereof etc.

Chinese patent CN1565465A discloses preparation of a kind of administrated by injection that contains the scutellarin active component and preparation method thereof.It is that active component scutellarin, medicinal basic and the pharmaceutical carrier of 98-102% formed that the feature of this patent mainly is by purity.

What all adopt in the injection preparation of foregoing invention is conventional injection preparation technology.Studies show that, because the scutellarin chemical property is active, very easily degraded inefficacy in the storage.Can in storage, degrade equally when common process is made injection or other liquid preparations, thereby cause content to descend, the general very difficult secular medicinal requirements that guarantees.There is not scutellarin injection products steady in a long-term to appear on the market before the present invention as yet.

Summary of the invention

Purpose of the present invention promptly attempts to obtain a kind of stable scutellarin drug solution, and reached more than a year and a half its storage period, and injection is used or the medicinal requirements of other approach to satisfy.The present invention has reached this purpose.

The present invention has used special stabilizing agent, and by PH regulator regulator solution pH value, adopts distinguished preparation method, has obtained gratifying effect, has made active constituent content and can stablize at least a year and a half scutellarin drug solution.This solution can satisfy the medicinal requirements of injection use or other approach.

The used stabilizing agent of the present invention comprises: EDTA-2Na, use amount 0.005%～1.0%; Carbonate or bicarbonate, use amount is an amount of; Vitamin C, use amount 0.05%～0.2%; Polyoxyethylene sorbitan monoleate, use amount 0.005%～0.5%; The aforementioned stable agent can be used separately, and combination also can suit.

As the PH regulator, drug solution of the present invention has also used sodium bicarbonate solution.

Distinguished preparation method is that the whole course of processing of the present invention has adopted the overall process nitrogen filled protection.

Studies show that, when using these stabilizing agents and PH regulator and adopting distinguished preparation method, beat allly obtained stable scutellarin drug solution, make the storage period stable content of this drug solution more than a year and a half.

The general preparation process of drug solution of the present invention is: under the overall process nitrogen filled protection, content is joined in the pharmaceutically acceptable water (as water for injection) of part preparation dosage (as 50%) up to the scutellarin raw material drug more than 98%, add bicarbonate solution and make till the dissolving fully; The stabilizing agent or its combination that add Sq are stirred and are made dissolving; Add water to full dose, regulate PH6.5～7.5 with sodium bicarbonate solution, promptly.

When as injection, increase the process of fine straining, embedding, sterilization.

By prescription of the present invention and manufacture method, scutellarin drug solution its storage period of preparing is more than a year and a half.

The applicant has carried out the anxious poison and the test of pesticide effectiveness with high-purity wild baicalin pharmaceutical composition of the present invention, has inquired into the safety of high-purity wild baicalin pharmaceutical composition and at the therapeutical effect of the treatment heart, cerebrovascular disease.Studies show that the scutellarin solution that the present invention prepares has reliable pharmacological action, its effect and safety are better than present conventional Breviscapini injection greatly, can be widely used in the treatment of the heart, cerebrovascular disease.

Detailed Description Of The Invention

Below demonstrate the present invention:

Embodiment 1:

Prescription: the former powder 40g of scutellarin

Polyoxyethylene sorbitan monoleate 4g

Vitamin C 20g

Sodium bicarbonate is an amount of

Add the injection water to 20000ml

Former powder 40g joins in the 10000ml water for injection with scutellarin, adds sodium bicarbonate solution and makes till the dissolving fully; Add 4g polyoxyethylene sorbitan monoleate, the stirring of 20g vitamin C 5 minutes, principal agent and adjuvant all dissolve; Add to the full amount of water for injection, regulate PH=7.05 with sodium bicarbonate solution, fine straining, embedding, sterilization promptly get the scutellarin injection.Content is 98.8%, and pH value is 7.0; Calculate after measured: it is 96.3% that the room temperature lucifuge is deposited a year and a half content, and pH value is 6.80.(overall process nitrogen filled protection)

Embodiment 2:

Prescription: the former powder 40g of scutellarin

EDTA-2Na 6g

Vitamin C 20g

Sodium bicarbonate is an amount of

Add the injection water to 20000ml

Former powder 40g joins in the 10000ml water for injection with scutellarin, adds sodium bicarbonate solution and makes till the dissolving fully; Add 6g EDTA-2Na, the stirring of 20g vitamin C 5 minutes, principal agent and adjuvant all dissolve; Add to the full amount of water for injection, regulate PH=6.95 with sodium bicarbonate solution, fine straining, embedding, sterilization promptly get the scutellarin injection.Content is 98.7%, and pH value is 6.91, calculates after measured: it is 96.5% that the room temperature lucifuge is deposited a year and a half content, and pH value is 6.87.(overall process nitrogen filled protection)

Embodiment 3:

Prescription: the former powder 40g of scutellarin

Vitamin C 10g

Sodium bicarbonate is an amount of

Add the injection water to 20000ml

Former powder 40g joins in the 10000ml water for injection with scutellarin, adds sodium bicarbonate solution and makes till the dissolving fully; Add the 10g vitamin C, stirred 5 minutes, principal agent and adjuvant all dissolve; Add to the full amount of water for injection, regulate PH=7.10 with sodium bicarbonate solution, fine straining, embedding, sterilization promptly get the scutellarin injection.Content is 98.5%, and pH value is 7.02, calculates after measured: it is 96.8% that the room temperature lucifuge is deposited a year and a half content, and pH value is 6.91.(overall process nitrogen filled protection)

The aforementioned stable data briefly gather as follows:

Sample source	Stable content
					0 month	June	December	18 months
	Embodiment 1	98.8％	98.2％	97.1％					96.3％
Embodiment 2					98.7％	98.3％	97.0％	96.5％

Sample source	Stable content
					0 month	June	December	18 months
	Embodiment 3	98.5％	97.8％	97.4％					96.8％

The applicant has carried out detailed pharmacology and pharmacodynamic study to high-purity wild baicalin pharmaceutical composition of the present invention, has inquired into the safety of high-purity wild baicalin pharmaceutical composition and at the therapeutical effect of the treatment heart, cerebrovascular disease.Result of study shows that high-purity wild baicalin has the good heart, cerebrovascular vascular applications prospect.The correlational study material is summarized as follows:

1.1 drug efficacy study

The scutellarin injection coagulates the cerebral tissue ischemia injury that intraluminal middle cerebral artery occlusion in rats causes to electricity the obvious treatment effect, 0.25～1mg/kg intravenous injection has tangible minimizing effect to the necrosis of cerebral tissue, simultaneously behavioristics's defective of improving animal of highly significant; Reperfusion injury also has the obvious treatment effect behind the focal cerebral ischemia that rat line bolt method is caused, and 0.25～1mg/kg intravenous injection obviously reduces the tissue necrosis rate, and behavioristics's defective is significantly improved; On the rat whole brain ischemia-reperfusion injury model, intravenous injection 0.25～1mg/kg scutellarin injection can prolong electroencephalogram extinction time behind the ischemia very significantly, shorten electroencephalogram recovery time and righting reflex recovery time after the perfusion again, and the anti-global brain ischemia reperfusion injury and protect of scutellarin injection obviously is better than positive drug nimodipine and Breviscapini injection, cerebral tissue moisture content behind the ischemia-reperfusion of scutellarin injection intravenous injection can the reduction simultaneously shows that the cerebral edema that this product causes ischemia-reperfusion has tangible preventive effect.Test of pesticide effectiveness result confirms, the intravenous injection of scutellarin injection to the experimental cerebral of animal and ischemia after again the tissue injury that causes of perfusion tangible prevention and therapeutical effect are arranged.

Find in the research aspect the anti-cerebral ischemia damnification mechanism: scutellarin injection various dose group and positive drug Radix Salviae Miltiorrhizae Injection have in various degree improvement to the blood fluidised form of rat brain microcirculation disturbance, showing as the hemocyte aggregation extent alleviates to some extent, become granular as the blood fluidised form by the stasis shape, or by granular band shape or the wire of becoming, or become wire by the dotted line shape, make meninges blood capillary site counting obviously increase (P＜0.05).The cerebral hemodynamic result of study shows, but scutellarin injection intravenous injection cerebral blood flow increasing amount is different with Radix Salviae Miltiorrhizae, its to the increase effect of cerebral blood flow greater than influence to periphery tissue blood flow amount.

Hemorheological result of study shows, the intravenous injection of high dose scutellarin injection can significantly reduce height cuts and hang down whole blood viscosity when cutting, and plasma viscosity is not then had influence, may improve erythrocyte deformability with it, suppress hemocyte and assemble relevant.The scutellarin injection also has the obvious suppression effect to the inductive platelet aggregation of AA, ADP and collagen-induced platelet aggregation then there is not the obvious suppression effect, rat arteriovenous thrombosis there is preventive effect, but fibrinogen content and plasmin activity are not had influence.Hemorheology Study result shows that scutellarin has good cardiovascular application prospect.

In addition, the stripped ileum longitudinal muscle that the scutellarin injection causes excitatory amino acid-glutamic acid shrinks more weak inhibitory action is arranged, and positive drug Ifenprodil has very strong inhibitory action to the stripped ileum longitudinal muscle contraction that glutamic acid causes, its dosage and effect are tangible positive correlation.The basilar artery ring experimental result that exsomatizes shows that the scutellarin of variable concentrations shrinks the inductive isolated rabbit basilar artery of KCl ring the dosage correlation inhibitory action.

Therefore, thus scutellarin injection anti-cerebral ischemia damaging action may with reduce whole blood viscosity, antiplatelet aggregation microcirculation improvement and brain blood for and to suppress the lipid oxidation enzyme system relevant with the calcium antagonism.Whether relevant with regard to its treating cerebral ischemia with the effect of antagonism excitatory amino acid, still await further research.

1.2 general pharmacology is learned research

This experiment is injected administration by vein is disposable, observes scutellarin to anesthesia Cor Canitis electricity, blood pressure, breathing and the general behavior of conscious mouse, the influence of spontaneous activity, and the pentobarbital sodium of observing this product and sub-threshold dose has or not collaborative syngignoscism.Experimental result shows, high dose (10mg/kg) scutellarin can cause that Canis familiaris L. one crosses property accelerated breathing and of short duration pressure reduction effect, and its maximum reducing effect occurs in after the administration 1 minute, just begin after 1 minute to recover gradually, return to the preceding level of administration in 5 minutes substantially.Middle dosage (3mg/kg) can reduce the systolic pressure of anesthesia Canis familiaris L., and low dosage (1mg/kg) does not all have obvious influence to electrocardio, breathing and blood pressure.But compare before each dosage group and the self administration, except that high dose group can obviously be accelerated to breathe after administration in 1 minute, the difference of comparing before other all observation indexs and the self administration that there are no significant.In view of scutellarin will be used for clinically in the intravenous drip administration from now on, peak concentration is injected the peak concentration of administration well below this any dosage after its administration, should be unable to occur one and cross property accelerated breathing and of short duration pressure reduction effect.Therefore we think that scutellarin intravenous drip administration do not have obvious influence to electrocardio, breathing and blood pressure.Scutellarin general behavior to conscious mouse when 1mg/kg does not obviously influence; The righting reflex score of animal has increase trend during 3～10mg/kg, especially at high dose 10mg/kg the righting reflex of animal is had more significantly influence; Scutellarin 3～10mg/kg also has more significantly influence to the pole-jump test of animal.Behind scutellarin injection 1～10mg/kg intravenous injection in 0.5～1 hour the spontaneous activity of mice reduce, returned to normal level in 2 hours substantially; And the pentobarbital sodium of this product and sub-threshold dose has collaborative syngignoscism, and its median effective dose ED50 (95% fiducial limit) to the test of barbital sodium subliminal hypnosis is 17.85 (9.46～36.89) mg/kg.

1.3 toxicologic study

1.3.1 acute toxicity test

1.3.1.1 mouse mainline acute toxicity test

Animal is evenly divided into groups by body weight, every group of each 10 mice of male and female, all at 81.2～2000.0mg/kg, dosage was in ratio setting in 1: 0.8 through the female male mice of prerun dosage range.The administration group is pressed the heavy every 0.4ml/10g volume intravenous injection of animal body, the normal saline solution that matched group deturs talis dosis, observe reaction of animals after the administration continuously, and record 1h, 2h, 4h, 1d, 2d, animal dead number in 3d to the 7d equal time, dead animal is carried out each main organs situation of anatomic observation, and surviving animals was dissected at the 7th day.Each dosage treated animal death toll is calculated other LD50 of male and female two individual characteies (95% fiducial limit) value with bliss method in the new drug statistical procedure.After the result shows the mouse mainline scutellarin, occur the paroxysmal tonic spasm at once, and with startling, spin until death, mouse diing time is all in 20 minutes.It is normal that survival mice was recovered after 2 minutes gradually.Existence and dead animal show no obvious abnormalities through each main organs of anatomic observation.The LD50 (95% fiducial limit) that calculates with the Bliss method is female to be 1280.0 (1143.13～1433.26) mg/kg, and male is 1469.05 (1302.23～1690.91) mg/kg.

1.3.1.2. mice oral administration gavage acute toxicity test

Animal is evenly divided into groups every group of each 10 animal of male and female by body weight.The dosage of female tom all is 6000mg/kg.Fasting is 4 hours before the zoopery.The administration group is pressed the heavy every 0.8ml/20g volume oral administration gavage of animal body, the matched group normal saline that deturs talis dosis, observe reaction of animals after the administration continuously, and animal dead number in record 4h, 1d, 2d, 3d to the 7d equal time, dead animal is carried out each main organs situation of anatomic observation, and surviving animals was dissected at the 7th day.After experimental result showed the oral scutellarin of mice, movable normal, not having faints from fear took place.Animal does not have death in 7 days.Each animal cervical vertebra dislocation in the 7th day is put to death, and shows no obvious abnormalities through each main organs of anatomic observation.Therefore the LD50 of female tom is all greater than 6000mg/kg.

1.3.1.3 mouse peritoneal injection acute toxicity test

Experimental result writhing response occurs after showing mouse peritoneal injection scutellarin, and prostrate motionless.Mouse diing time is all in 48 hours.Not having faints from fear takes place.It is normal that survival mice was recovered after 3 days gradually.Existence and dead animal show no obvious abnormalities through each main organs of anatomic observation.The LD50 (95% fiducial limit) that calculates with the Bliss method is female to be 2240.00 (2000.47～2508.21) mg/kg, and male is 2284.45 (2064.36～2528.22) mg/kg.

1.3.1.4 Beagle dog studies on acute toxicity

This test is injected heavy dose of scutellarin by giving clear-headed Beagle Canis familiaris L. disposable vein, the observation sample scutellarin is to the acute toxic reaction of beagle dog, grope the approximate lethal dose of scutellarin, judge histoorgan or system that toxic reaction may relate to, for clinical application provides reference.The experimental study result shows: scutellarin injection 166.7,375.0 and 843.8mg/kg dosage respectively after the administration about 4min sialorrhea all takes place, then vomiting, diarrhea, lie prostrate motionlessly, the low dosage Canis familiaris L. recovers behind 20min gradually, 1 hour basic recovery normally; Slow down 375.0mg/kg breathe to deepen during the about 25min of Canis familiaris L., vomiting reaction strengthens, and symptom alleviates gradually after 1 hour, and 3 hours basic recovers normal.843.8mg/kg sialorrhea, vomiting, diarrhea take place for about 4min after the Canis familiaris L. administration, the ground that crouches can not be uprightly, orthocolosis and opisthotonus, strong vomiting took place in the time of 8 minutes move, the platycoria death of the 15min left and right sides is dissected each main organs and is shown no obvious abnormalities.Further give 375 and the middle dosage 562.5mg/kg of 843mg/kg, vomiting after the administration, crouching ground can not be upright, orthocolosis, opisthotonus, strong vomiting action takes place during 8min, and the platycoria death of the 15min left and right sides is dissected each main organs and is shown no obvious abnormalities.Give 5# and 6# Canis familiaris L. with scutellarin 470mg/kg again, the 6# Canis familiaris L. vomits, gatism, and it is motionless to fall down to the ground, extremity stretch, opisthotonus, and whole body is twitched and is trembled, grit one's teeth, binocular magnifies, and relaxes during breathing and when rapid, saliva foam and urine are dark green, breathe slightly behind about 20min to show steadily, but still have the tic phenomenon, and the increase of twinkling, muscle trembles and the vomiting reaction enhancing about 1 hour, and is dead about 20` divided in 2 hours, dissects the whole body internal organs and show no obvious abnormalities.Begin vomiting, fecal incontinence after the administration of 5# Canis familiaris L., but medicine is injected and still can be stood after finishing.Lie prostrate behind the 5min, recover normal after 4 hours gradually.Surviving animals is in 14 day observation period subsequently, behavioral activity, outward appearance sign, diet, body weight, secretions and Excreta color and character are all normal, comparing before the every index of electrocardio and the administration does not also have significant change, dissects huge inspection at last and does not find that each internal organs and tissue have tangible pathological change.Therefore the general fatal dose of the clear-headed beagle Canis familiaris L. of scutellarin intravenous injection single administration is about 470mg/kg.

1.3.2 long term toxicity test

1.3.2.1 rat long term toxicity test

The SD rat is adopted in this experiment, and 20 every group, male and female half and half are divided into matched group and 5,15, three administration groups of 50mg/kg/d, intraperitoneal injection 3 months.

Rat lumbar injection scutellarin is 3 months long term toxicity test results show, the inspections such as general situation (outward appearance sign, behavioral activity, feces character, diet drinking-water, glandular secretion, body weight etc.), hematology, serum biochemistry, routine urinalysis, system's postmortem, organ coefficient and histopathology of animal be there is no the overt toxicity reaction relevant with medicine.Inspections such as the hematology in 4 weeks of drug withdrawal, serum biochemistry, routine urinalysis, system's postmortem and organ coefficient also there is no the overt toxicity reaction relevant with medicine.

The continuous lumbar injection scutellarin of rat 3 months, its nontoxic dose are 50mg/kg/d (be equivalent to rat effective dose 100 times).

1.3.2.2 Beagle Canis familiaris L. long term toxicity test

The beagle dog is adopted in this experiment, is divided into matched group and scutellarin 2,7, three administration groups of 25mg/kg/d, every group of 8 animals, male and female half and half, every day, intravenous drip was administered once, and administration is 6 days weekly, continued for 13 weeks, put to death the half animal and check that comprehensively drug withdrawal was observed for 3 weeks and seen its recovery situation then.

Administration is 13 all beagle dog long term toxicity test results show, intravenous drip scutellarin injection, high dose group has in 2 animals administration process in early days, repeatedly (totally 6 times) sialorrhea occurs, feel sick, symptoms of emesis, part high dose group animal when drip velocity is fast, have after administration finishes of short durationly lie prostrate, when standing shank tremble, body-sway motion, astasia phenomenon, return to normal after tens of approximately seconds, it is inferior that 9 examples take place in the process of the test altogether, and the drip velocity that slows down is like preventing that phenomenon takes place here.Administration mid-term and latter stage, administration group and control animals platelet count have significant difference statistically, but both there be not dose-effect relationship, also irregular following, in the entire test, the all upper and lower fluctuations in normal range of matched group and administration group cause so this difference should be test error, and are irrelevant with sample.

Except that the sialorrhea and vomiting of high dose group, 13 weeks of administration are to other index of animal, as situation (comprising outward appearance sign, behavioral activity, feces character, diet drinking-water, administration local response, body weight etc.), hemogram, liver function, serum biochemistry there is no the obviously toxic reaction relevant with inspections such as ion, routine urinalysis, the huge inspection of internal organs, organ coefficient and myelopathy Neo-Confucianism with medicine.Histopathology result shows, high dose group has 2 animals to occur obviously and the reaction of the gastrointestinal toxicity of drug-associated: atrophic gastritis, the mucosa top layer is slight downright bad and have and come off, edema still arranged under the gastric mucosa, chronic duodenitis and ileitis are arranged, intestinal mucosa is all seen partial necrosis and is come off, ileum lymphoid tissue severe hypertrophy, the necrosis of animal mucous membrane of colon epithelium is replaced by the mucoid exudate, a large amount of hypertrophy of enteraden goblet cell.Cavity sample degeneration not of uniform size appears in granular degeneration of hepatic tissue part cell and aorta.

The inspections such as the general situation of animal, hemogram, liver function, serum biochemistry and ion, routine urinalysis, the huge inspection of internal organs, organ coefficient and myelopathy Neo-Confucianism in 13 all 3 weeks of drug withdrawal of administration also there is no the overt toxicity reaction relevant with medicine

Beagle dog continuous intravenous infusion scutellarin 13 weeks of injection, consider that 25mg/kg has obviously gastrointestinal tract, behavioristics and the histologic reaction relevant with medicine, so think this dosage for this test in the dosage of toxic reaction, but the toxic reaction that causes under this dosage is not observed on the convalescent period animal, is the reversibility pathological change.Its non-toxic is 7mg/kg/d, (be equivalent to rat drug effect dosage 14 times).

1.3.3 specific safety test

The scutellarin injection does not have the obvious stimulation effect to rabbit auricular vein blood vessel; Injection has slight stimulation to the rabbit quadriceps femoris; Hypersensitive test result to rabbit erythrocyte hemolytic test and Cavia porcellus is all negative; The scutellarin injection is also negative to the passive cutaneous anaphylaxis test result of rat.

1.3.4 mutagenicity test

Designing requirement by " new drug (Western medicine) preclinical study guideline compilation ", selecting histidine defect type Salmonella typhimurium (S.typhimurium) TA97, TA98, TA100, TA102 and TA1535 for use is indicator strain, has detected the mutagenicity of scutellarin injection.Experiment is not adding and is not adding under metabolism activation system (S9) condition and carry out, and adopts dressing plate to mix method.The result shows that the scutellarin injection adding and not adding under the condition of S9, does not have mutagenicity to Salmonella typhimurium.

Adopt Chinese hamster ovary fibroblast (CHO) in vitro tests method, detected the mutagenicity that is tried thing, test establishes 58,116 and three dosage groups of 232mg/ml.The result shows: adding S9 and not adding the Chinese hamster ovary celI chromosomal aberration of all not bringing out cultivation under the condition of S9 metabolism activation system, illustrate that this medicine does not have the aberration inducing effect to the Chinese hamster ovary celI chromosome.

Use mouse bone marrow cells micronucleus test method the scutellarin injection is carried out mutagenicity research.The result shows: (female: 256～1024mg/kg at set dosage range, male: in 293.8～1175.2mg/kg), the micronucleus of NIH mice is brought out rate and negative control group does not more all have significant difference (P＞0.05), show that being tried thing scutellarin injection does not have the effect of bringing out bone marrow polychromatic erythrocyte micronucleus to the NIH mice.

Result of the test shows that scutellarin does not have mutagenicity.

1.3.5 reproduction toxicological test

1.3.5.1 rat II section test

The SD pregnant rat at sensitive period to teratogenic agent (conceived the 6th～15 day) through intravenous injection 10,35 and 120mg/kg scutellarin, continuous ten days.Experimental result shows that female sodoku appears at the 120mg/kg group, and development toxicity also appears at the 120mg/kg group, does not see because of drug-induced monster is young to occur under 120mg/kg dosage, and scutellarin is the medicine with embryo, the young teratogenecity of tire.

1.4 pharmacokinetic

According to the drug efficacy study result, adopt 1,3 and three dosage groups of 9mg/kg, the rat vein administration, after administration 0,1,2.5,5,15,30,60,90,120 and 180min get blood, detect blood Chinese medicine concentration; With after the administration 5,30,60 and each 6 animal of 90min high dose group, make tissue distribution and detect; With 9mg/kg dosage 6 rats are carried out excretion test.The result shows, the drug-time curve of quiet notes 1,3 of rat and 9mg/kg, through process of fitting treatment, the feature that meets linear two-compartment model, the half-life t1/2a that calculates by two-compartment model is respectively 1.52,1.47,3.43min, t1/2b is respectively 53.16,51.08,45.63min, and apparent volume of distribution is 4.1136,1.4271,3.2218L/kg, and clearance rate is 0.0536,0.0194 and 0.0489L/min/kg.Scutellarin area under the drug-time curve AUC proportional increase when dosage increases to 9mg/kg by 1mg/kg, correlation coefficient is 0.997, shows linear pharmacokinetics feature in the test dose scope.5min after the intravenous injection promptly has distribution in each internal organs, wherein liver, kidney, small intestinal content are higher; The concentration of other each tissues is lower.Each time point, the drug level of ovary is all apparently higher than testis.Remedy,tissue's substrate concentration descends also with plasma concentration, and respective synchronization descends.Behind the quiet notes scutellarin of the rat 9mg/kg, the original shape medicine of after administration, from urine, discharging in 2 hours be 1.42%, 8 hour be 2.60%, 24 hour be that the homaluria total amount reaches dosage in 3.08%, 48 hour 3.38%.48 hours excretion is 1.97% in the feces.The excretory original shape medicine total amount of urine and excrement is 5.35% of a dosage.After intravenous injection this product, the part medicine can be secreted in bile by liver, and the peak value of bile excretion is after the administration in 0-2 hour, and 24 hours total excretion is about 9.52% of dosage, greater than feces Chinese medicine output.Show by bile secretion after intestinal is drained is another elimination approach of this product, may have a certain amount of liver sausage circulation simultaneously; Also may be because the unstability of medicine itself makes medicine change other chemical compounds in feces.This product protein binding rate is medium, and when 0.024,1.804,3.608 μ g/mL concentration, combination rate is respectively 58.6,62.8,58.8%.The intravenous blood plasma pharmacokinetic properties of Canis familiaris L. also meets the linear two-compartment model of eliminating, 1,3 and the elimination half-life t1/2a of 9mg/kg dosage group is respectively 4.26,3.93,1.48min, t1/2b is 70.43,43.14,31.92min, area under the drug-time curve AUC is proportional increase when dosage increases to 9mg/kg by 1mg/kg, is linear kinetics and eliminates (r=0.9996).

Claims (3)

1. a high-purity wild baicalin injection is characterized in that this injection contains high-purity wild baicalin, stabilizing agent and pH regulator agent, adopts the overall process nitrogen filled protection during preparation:

High-purity wild baicalin content is more than 98%;

Stabilizing agent is that sodium bicarbonate and vitamin C are united use, or sodium bicarbonate, vitamin C and EDTA-2Na unite use, or sodium bicarbonate, vitamin C and polyoxyethylene sorbitan monoleate are united use;

The consumption of stabilizing agent is respectively: EDTA-2Na, 0.005～1.0%; Vitamin C, 0.05%～0.2%; Polyoxyethylene sorbitan monoleate, 0.005～0.5%;

The pH regulator agent is a sodium bicarbonate, and pH is 6.5～7.5.

2. according to the described high-purity wild baicalin injection of claim 1, it is characterized in that:

Prescription: the former powder 40g of scutellarin

Polyoxyethylene sorbitan monoleate 4g

Vitamin C 20g

Sodium bicarbonate is an amount of

Add the injection water to 20000ml

Method for making: former powder 40g joins in the 10000ml water for injection with scutellarin, adds sodium bicarbonate solution and makes till the dissolving fully; Add 4g polyoxyethylene sorbitan monoleate, the stirring of 20g vitamin C 5 minutes, principal agent and adjuvant all dissolve; Add to the full amount of water for injection, regulate PH=7.05 with sodium bicarbonate solution, fine straining, embedding, sterilization promptly get the scutellarin injection; The overall process nitrogen filled protection.

2, the described high-purity wild baicalin injection of claim 1 is characterized in that:

Prescription: the former powder 40g of scutellarin

EDTA-2Na 6g

Vitamin C 20g

Sodium bicarbonate is an amount of

Add the injection water to 20000ml

Method for making: former powder 40g joins in the 10000ml water for injection with scutellarin, adds sodium bicarbonate solution and makes till the dissolving fully; Add 6g EDTA-2Na, the stirring of 20g vitamin C 5 minutes, principal agent and adjuvant all dissolve; Add to the full amount of water for injection, regulate PH=6.95 with sodium bicarbonate solution, fine straining, embedding, sterilization promptly get the scutellarin injection; The overall process nitrogen filled protection.

4. according to the described high-purity wild baicalin injection of claim 1, it is characterized in that:

Prescription: the former powder 40g of scutellarin

Vitamin C 10g

Sodium bicarbonate is an amount of

Add the injection water to 20000ml

Method for making: former powder 40g joins in the 10000ml water for injection with scutellarin, adds sodium bicarbonate solution and makes till the dissolving fully; Add the 10g vitamin C, stirred 5 minutes, principal agent and adjuvant all dissolve; Add to the full amount of water for injection, regulate PH=7.10 with sodium bicarbonate solution, fine straining, embedding, sterilization promptly get the scutellarin injection; The overall process nitrogen filled protection.