CN101480391B - Application of sophocarpine in preparing medicament for treating disease induced by coxsachie A virus - Google Patents

Abstract

The invention discloses an application of sophocarpine or officinal salt thereof in the preparation of medicine for treating myocarditis or brain fever caused by Coxsaickie Viruses A. The sophocarpine or the officinal salt thereof can effectively prevent the biosynthesis of the Coxsaickie Viruses A, enter cells to play an antivirus role, improve the cure rate and reduce the death rate, and moreover, the sophocarpine or the officinal salt thereof has no obvious toxic effect for the cells, has abundant medicine sources of sophocarpine, simple and convenient preparation and low cost, and is suitable for a commercial process.

Description

Sophocarpine is treated by the application in the medicine of Coxsackie A disease disease that poison causes in preparation

Technical field:

The present invention relates to field of medicaments, relate in particular to the application of sophocarpine or its officinal salt, particularly a kind of sophocarpine is treated by the Coxsackie A disease application in myocarditis or the meningitic medicine that poison causes in preparation.

Background technology:

The viral myocarditis showed increased is mainly caused by enterovirus in recent years.Enterovirus is mainly provincialism or diffusing point is popular, and it is main that family's contact method infects.Enterovirus is the infectiousness of height to Susceptible population, and the child is the most responsive crowd, in comprising pharyngeal gastrointestinal epithelial cells and lymphoid tissue, breeds.In child who lacks immunity or adult, virus further is diffused into blood circulation, causes viremia.This moment, heating appearred in patient, and at this moment virus gets into whole body reticuloendothelial cell and lymphoid tissue continuation breeding on the one hand, also can invade responsive organ or tissue such as spinal cord, brain, meninges, cardiac muscle and skin etc. and cause special clinical manifestation.

Many viruses can cause myocarditis, have to think enterovirus more than 24 kinds is arranged (like Coxsackie virus (Coxsackic virus), echovirus (Echo VIRUS), poliovirus (because the vaccine of the scorching virus of grey marrow, not seeing has morbidity).Coxsackie virus is divided into two groups, and CBV has CVB _1-6Type is CVB wherein _1-5All can cause a disease, especially CVB _2-4Type be viral myocarditis main diseases because of.The person is also quite a lot but the Coxsackie A disease poison causes the viral myocarditis.Coxsackie A disease poison (CVB _{The 1-24 type}, CVB wherein _4,7,9,10,23Be prone to cause a disease, especially 7 types, 9 types cause myocarditis, pneumonia and meningitis easily.Echovirus also can cause viral myocarditis for enterovirus equally.And the medicine of anti-COXB still not yet in effect so far, COXA and Echo virus.For seeking the active drug of antagonism COXB, COXA and Echo virus, we only filter out Radix Sophorae Flavescentis simply through experiment from numerous Chinese medicine.Its low toxicity of effective ingredient sophocarpine-Sophocarpine in the Radix Sophorae Flavescentis and obvious antagonism COXB, COX are arranged _AAnd the effect of Echo virus.Radix Sophorae Flavescentis is one of time-honored conventional medicament of China, firstly appears the medical literature Shennong's Herbal the earliest in China, has the effect of heat clearing and damp drying, wind dispelling insecticide.Cure mainly trusted subordinate's feeling of fullness flatulence, lump in the abdomen, jaundice etc.China's pharmacopeia version in 1997 and some areas drug standard will " Sophora alopecuroides stream has soaked sheet " have recorded and have cured mainly bacillary dysentery and enteritis " Chinese herbal medicine (middle volume) ", in August, 1976 front page (494 ~ 495).Zoopery in recent years proves that Radix Sophorae Flavescentis alkaloid has inhibition, eases pain and fall wet effect to nervus centralis, ability human body immunity improving power, and antitumaous effect is arranged.The Radix Sophorae Flavescentis extract alkaloid can be treated hepatitis B, improves clinical symptoms and liver function, the effect that part patient HBeAg is turned out cloudy.Having document record Radix Sophorae Flavescentis alkaloid to have relievings asthma and the antiarrhythmic effect.Its low toxicity of effective ingredient sophocarpine (Sophocarpine) in the Radix Sophorae Flavescentis and the effect of obvious antagonism COXB, COXA and echovirus is arranged.Be published in my one in the granted patent 99119850.6 about sophocarpine antagonism COXB virus and medication preparation thereof.In addition, the present inventor also found sophocarpine to anti-coronavirus (SARSvirus) correlation technique content at another in the granted patent 03151269.0.The present inventor finds that also sophocarpine can resist echovirus (Echo virus) adenovirus (Adeco-virus) and influenza B virus (influenza virus B); It can be to RNA virus resisting; Again can be to resisting DNA virus; Therefore it is a kind of broad-spectrum antiviral medicament, and the correlation technique content discloses (open) to some extent in another patent application 200410089431.6 of the applicant.

Summary of the invention:

The object of the present invention is to provide a kind of sophocarpine to treat by the application in the medicine of Coxsackie A disease disease that poison causes in preparation, described this sophocarpine or its officinal salt will solve does not have the malicious institute of effective Drug therapy Coxsackie A disease to cause myocarditis or meningitic technical problem in the prior art.

The invention provides sophocarpine or its officinal salt and treat by the application in Coxsackie A disease caused myocarditis of poison or the meningitic medicine in preparation, wherein, the structural formula of described sophocarpine is (I)

Further, the officinal salt of described sophocarpine is bromate or the hydrochlorate of sophocarpine or the fluorate of sophocarpine of sophocarpine.

Further, the dosage form of the medicine of described sophocarpine or its officinal salt is any dosage form of described pharmaceutical dosage form for medically approving.

Further, the pharmaceutical dosage form of described sophocarpine or its officinal salt is powder or injection or capsule or tablet or oral liquid.

The method for preparing of sophocarpine of the present invention comprises the steps:

(1) Herba Sophorae alopecuroidis is pulverized, the aqua ammonia of adding 5%～10% is moistening, extracts through the chloroform reflux, adds the extraction of 7%～15% sulphuric acid, obtains total alkaloids.

(2) total alkaloids adds 5%～10% hydrochloric acid extraction sophocarpine then through extracted in toluene.

(3) regulate PH5～10, sucking filtration again with acetone recrystallization, obtains the sophocarpine elaboration

(4) the sophocarpine elaboration is dissolved in the injection water, filters, assay is regulated PH4～4.5.

(5) through 0.65 microporous filter membrane filling and sealing.Can make injection in 30 minutes through 100 ℃ of sterilizations of flowing steam.

Sophocarpine of the present invention can effectively stop the biosynthesis (comprising protein synthesis) of Coxsackie A disease poison and get into performance antivirus action in the cell.

In cardiac muscle biopsy, be difficult for isolating any virus because the back takes place viral myocarditis, specific antigen is also difficult finds that the viral myocarditis cause of disease mainly relied on Serological testing in the past, like specificity neutralizing antibody and IgM.But owing to antibody can be passed in time and reduces, disappears.Thereby sensitivity that serology detects after acute stage and specificity reduction; In recent years owing to adopted in situ hybridization and polymerase chain reaction (Polymersae Chain Reaction-PCR) etc. to detect the viral gene in endomyocardial biopsy and the corpse; Find that COXA virus is much, and affirmed that COXA virus also is one of cause of disease of viral myocarditis.

COXA papova (coxsaicki A-virus) divides 24 types, and wherein 7 types, 9 types are prone to cause pathological changes such as viral myocarditis, pericarditis and viral meningitis.It is the important cause of disease of infant viral myocarditis and encephalitis.CVA ₂₃Type can cause adult's fulminant myocarditis (sometimes with the influenza virus double infection); Even cause severe arrhythmia sudden deaths such as paroxysmal auricular tachycardia, room Piao, atrial fibrillation, the ventricular premature contraction that takes place frequently, atrioventricular block, acute heart failure and contraction cardiac insufficiency.The COXA papova can cause that meningitis (generally claiming aseptic meningitis) onset is medium degree heating; Sometimes similar cold symptoms often with headache, nauseating, vomiting, has muscular soreness occasionally; Stiffness of nape and back appears in weight person, and the cerebrospinal fluid cell number increases and protein increases.As being difficult for differentiating with the epidemic encephalitis type B case sometimes in summer.Can in cerebrospinal fluid, find the COXA viral gene with the PCR method, or isolated viral.Shanghai once was separated to COXA in 1964 from the cerebrospinal fluid of aseptic meningitis ₉, follow-up period also was separated to COXA from the cerebrospinal fluid of aseptic meningitis in 1960～1970 years ₉, COXB ₄Virus etc.

For this reason, we have further done research to the effect of sophocarpine antagonism COXA virus.Observed sophocarpine anti-COXA on the Hela cell ₉Effect, observed sophocarpine that crystal violet competent cell staining measures variable concentrations to infecting COXA ₉The protective effect of Hela cell survival rate, also observed tetramethyl azo azoles salt (MTT) method and measured sophocarpine infecting COXA ₉The metabolic protective effect of Hela cellular energy of virus.Find that all sophocarpine has obvious antagonism COXA ₉The effect of virus and to the protective effect of infection cell.

Pass through COXA ₉The viremia model of the Balb/c mice of viral infection, myocarditis model and encephalitis model are observed sophocarpine in vivo to COXA ₉The antagonism of virus.The result finds that it is to COXA ₉The inhibited proliferation of virus reaches 99%, and can obviously alleviate the variation of the histopathology of myocarditis and brain, and the obvious treatment effect is arranged.

The present invention and existing compared with techniques, its effect are actively with tangible.Sophocarpine of the present invention or its officinal salt can effectively stop the biosynthesis of Coxsackie A disease poison, and get into performance antivirus action in the cell, can improve more rate of treatment, reduce mortality rate.And not having a significant cytotoxicity effect, sophocarpine medicine source is abundant, and prepares easyly, with low cost, is applicable to suitability for industrialized production.

Description of drawings:

Fig. 1 is the photo of Balb/c mice viral myocarditis due to the COXA9 virus, large stretch of cardiac muscle of visible visceral pericardium and myocardium inflammatory disorders.

Fig. 2 has shown that sophocarpine treatment group (10mg/kg) visceral pericardium inflammation obviously alleviates a small amount of inflammatory kitchen range of myocardium.

Fig. 3 has shown that sophocarpine treatment group (20mg/kg) visceral pericardium cardiac muscle and myocardium do not see obvious inflammation.

Fig. 4 is the photo of the ependymitis of Balb/c mice due to the COXA9 virus.

Fig. 5 has shown that sophocarpine treatment group (10mg/kg) ependymitis pathological changes alleviates.

Fig. 6 has shown that sophocarpine treatment group (20mg/kg) do not see ependymitis.

The specific embodiment:

Embodiment 1

The method for preparing of sophocarpine comprises the steps:

(1) Herba Sophorae alopecuroidis is pulverized, the aqua ammonia of adding 5%～10% is moistening, extracts through the chloroform reflux, adds the extraction of 7%～15% sulphuric acid, obtains total alkaloids.

(2) total alkaloids adds 5%～10% hydrochloric acid extraction sophocarpine then through extracted in toluene.

(3) regulate PH5～10, sucking filtration again with acetone recrystallization, obtains the sophocarpine elaboration

(4) the sophocarpine elaboration is dissolved in the injection water, filters, assay is regulated PH4～4.5.

(5) through 0.65 microporous filter membrane filling and sealing.Can make injection in 30 minutes through 100 ℃ of sterilizations of flowing steam.

The test of the external anti-COXA9 virus of embodiment 2 sophocarpine

Seed culture of viruses preparation: COXA ₉Strain is provided by Zhao Shu of Kunming Institute of Medical Biology of the Chinese Academy of Sciences.Get and cultivate the Hela cell that grew up to monolayer on the 4th day, wash secondary, every bottle of cell inoculation 0.3ml10 with the RPMI-1640 culture medium of no calf serum ^-2TCID COX A ₉37 ℃ of absorption of virus 1 hour, the supernatant that inclines, adding 10ml does not have calf serum RPIM-1640 culture medium, 37 ° of 5%CO ₂Hatch cultivation, treat the complete pathological changes of cell after, take out cell and multigelation 6 times.Collection is gone down to posterity as new inoculum.After the continuous passage like this 4～5 times, last freeze thawing thing collection is put-30 ℃ store for future use.

The trace cell culture method is measured virus titer: viral freeze thawing liquid is done continuous 10 times of dilutions with no calf serum RPIM-1640 culture medium.From 10 ^-1～10 ^-8Totally 8 dilution factors, on 40 hole microdetermination plates, each dilution factor is done 4 multiple holes, and every hole amount of splashing into is 25 μ l, and restock 25 μ l in every then hole do not have calf serum RPIM-1640 culture medium, and adding concentration at last is 3 * 10 ⁵The Hela cell suspension of/ml, every hole 50 μ l (calf serum is 4% in the culture medium) are done the normal cell contrast simultaneously, and plate is put 37 ℃, contain the 5%CO2 incubator and cultivate the 4th day sentence read result.50% above cytopathy occurring with the high dilution of virus is the titre terminal point.

The trace cell culture method is measured COXA ₉Virus titer

The viral dilution degree	10 -1	10 -2	10 -3	10 -4	10 -5	10 -6	10 -7	10 -8	The cell contrast
										Cytopathy hole/total cell hole count	4/4	4/4	4/4	4/4	4/4	3/4	1/4	0/4	0/4

Press the Reed-Muench formula and calculate, virus titer is 10 ^-6.5TCID ⁵⁰(Tissue culture infections dose).

Embodiment 3 sophocarpine are to the toxicity test of Hela cell

To be 100mg/ml (Renji Hospital Attached to Medical College of Shanghai Jiaotong Univ. provides the rich pharmaceutical factory of standing grain to make) be diluted to 1% liquor strength with no calf serum RPIM-1640 culture fluid to sophocarpine that (10000 μ g/ml are mother solution, the sterile cryogenic preservation.

With concentration is that 1% sophocarpine injection is diluted to variable concentrations (the continuous doubling dilution to 1 from 1: 2: 1024) add the hole respectively with no calf serum RPIM-1640 culture fluid; Each dilution factor is that cultivated 4 days in 4 multiple holes (and establishing normal control); Every day, observation of cell toxicity was calculated half toxic dose (TD ₅₀) and maximal non-toxic dosage (TD ₀).

Sophocarpine is to the toxicity of Hela cell

The sophocarpine dilution factor

1∶2

1∶4

1∶8

1∶16

1∶32

1∶64

1∶128

1”256

1∶512

1∶1024

Cytopathy

+

±

-

-

-

-

-

-

-

-

Annotate: when sophocarpine is diluted to 1: 8 when following, promptly concentration is when 312 μ g/ml are following, to Hela cell avirulence.

TD ₅₀Be 624 μ g/ml, TD ₀Be 312 μ g/ml.

Embodiment 4 sophocarpine injection resist COXA on the Hela cell ₉The effect of virus

(1) micromethod is measured sophocarpine anti-COXA on the Hela cell ₉The virus test

Sophocarpine is anti-COXA on the Hela cell ₉Effect (the COXA of virus ₉Virus concentration is 100TCID ₅₀)

The sophocarpine dilution factor

1∶8

1∶16

1∶32

1∶64

1∶128

1∶256

1∶512

1∶1024

Cytopathy

-

-

-

-

-

-

±

+

Annotate: when dosage 1; 8 promptly all have tangible anti-COXA during the sophocarpine injection below the 312 μ g/ml ₉The effect of virus still has anti-COXA when 1: 512 (i.e. 5 μ g/ml) ₉The effect of virus.

Embodiment 5 sophocarpine are to COXA ₉Viral infection Hela cell the protective effect of energy and survival rate measure

MTT trace detection method (tetramethyl azo azoles salt) is measured sophocarpine to COXA ₉Virus is to infecting the metabolic protective effect of Hela cellular energy.

Crystal violet staining assay (crystal violet): measure sophocarpine to COXA ₉Virus is to infecting the protective effect of Hela cell survival rate.

Measure sophocarpine to infecting COXA ₉The energy metabolism of the Hela cell of virus and the protective effect of survival rate require COXA ₉The concentration of virus is carried out the mould rope.

Getting the Hela cell of cultivating the 4th day and process suspension, abandon culture fluid, use no calf serum RPIM-1640 culture fluid Xian twice, is 5 COXA with infection multiplicity ₉37 ℃ of absorptions of viral suspension are abandoned viral liquid 2 times; Again with twice of no calf serum RPIM-1640 culture fluid Xian; The calf serum RPIM-1640 culture fluid 200 μ l that add sophocarpine 100,50,25,12.5,6.25,3.125,1.563 μ l/ml then respectively; If medicine matched group (sophocarpine 200 μ l/ml), virus control group, cell matched group, 37 ° of 5%CO ₂Hatched 15 hours.

COXA ₉The mould rope of virus concentration

Method

COXA 9Virus-infection multiplicity

0

2

2.5

4

5

10

20

MTT colorimetry (D570nm)

0.495 ±0.026

0.431 ±0.067

0.400 ±0.433

0.387 ±0.058

0.370 ±0.044

0.340 ±0.035

0.334 ±0.026

Violet staining (OD600nm)

1.580 ±0.091

1.464 ±0.12

0.448 ±0.244

1.309 ±0.065

1.290 ±0.140

1.252 ±0.133

1.125 ±0.122

Annotate: the variation of measured value under the different virus infection multiplicity, two kinds of distinct methods results are like table.The result shows that with the rising of virus concentration, the cellular metabolism rate that mtt assay records is low more, and both are negative correlation (r=-0.805), and it is also low more that crystal violet staining assay records cell survival rate, and both are negative correlation (r=-0.883) linearly also.

According to result with the patrix rope, selected COXA ₉The viral infection plural number is that 5 o'clock (MOI=5) surveys the sophocarpine injection to COXA ₉The Hela cytoprotection of viral infection is appropriate.

Sophocarpine is to COXA ₉The protective effect of the Hela cell of viral infection

Sophocarpine concentration	COXA 9The viral infection plural number is 5 o'clock (MOI:5)
			MTT colorimetry (D570nm)	Crystal violet staining assay (OD600nm)
	100(μg/ml)	0.498±0.018			2.083±0.42
50(μg/ml)			0.489±0.02	1.801±0.148
	25(μg/ml)	0.483±0.045			1.707±0.148
12.5(μg/ml)			0.473±0.041	1.600±0.173
	6.25(μg/ml)	0.428±0.026			1.57±0.115
3.125(μg/ml)			0.401±0.022	1.514±0.203
	1.563(μg/ml)	0.398±0.054			1.500±0.101
The cell contrast			0.518±0.026	2.193±0.282
	Virus control	0.351±0.012			1.134±0.060
The medicine contrast			0.500±0.033	2.000±0.163

Shown in the last table:: work as COXA ₉The viral infection plural number is 5 o'clock, and sophocarpine all has protective effect to the Hela cell between 1.563～100 μ g/ml.Energy anabolism, cell survival number average increase with the rising of drug level, are proportionate, and r is respectively 0.913 and 0.985.

(1) the medicine matched group has been compared very notable difference P＜0.001 with the virus control group.

(2) the cell matched group has been compared notable difference P＜0.001 with the virus control group.

(3) medicine matched group and cell matched group no significant difference P＞0.05.

Explain that sophocarpine is to COXA ₉The survival rate effect of adequately protecting of the energy anabolism of the Hela cell of viral infection and cell.

Embodiment 6 sophocarpine are to COXA ₉The intravital antivirus action of virus animal model

The virus multiplication inhibition test is measured sophocarpine in vivo to COXA ₉The inhibitory action of virus.

COXA ₉Behind the viral infection Hela cell, virus is bred in cell, because cell is topped with agarose, virus can only be through cell-cell by way of propagation.So cytoclasis, the cracking of regional area property occur, cell death is after fixed and stained, and the destruction region presents plaque.Each plaque is formed by a viral infection unit, so virus titer can be accurately measured in the plaque test.Plaque reduces experiment can reflect accurately that sophocarpine suppresses COXA ₉The propagation degree of virus.This experiment is adopted virus multiplication to suppress method (being that plaque reduces test) and is observed sophocarpine in vivo to COXA ₉The inhibitory action of virus.

Anti-COXA in embodiment 7 bodies ₉The effect of virus

Suppress method (plaque forming method) through cell culture method with virus multiplication and measure sophocarpine COXA ₉The inhibitory action of virus multiplication.

One .COXA ₉The viremia animal model making

The sophocarpine injection: He Feng pharmaceutical factory in Shanghai provides specification: 200mg/2ml, lot number: 030827.

COXA ₉Viral source: Kunming Institute of Medical Biology, Zhao Shudong provides.

Laboratory animal: the Balb/c mice in 3～4 ages in week, must be provided by triumphant laboratory animal company limited by the Sino-British joint western pul in Shanghai.

COXA ₉Viremia Animal Model Making: get 20 of Balb/c mices.

A group: COXA ₉10 on viral infection model

B group: 10 of sophocarpine injection for treating groups

Two groups each is at intraperitoneal inoculation 0.1ml100TCID ₅₀COXA ₉Virus.B organizes behind virus inoculation 2 hours beginnings with sophocarpine injection 10mg/kg/ day by tail vein injection physiological saline solution 0.1ml two/day 2 hours beginnings after the A winding kind, branch secondary/day tail vein injection.All get blood under aseptic condition, extracing eyeball of mouse on the 3rd day after the inoculation for two groups, put into sterile test tube, leave standstill and treat that serum separates out; Every group of 10 mices are got 0.1ml serum mixed in equal amounts; Add mycillin, concentration is 2000 μ/ml, puts 4 ℃ of refrigerators 2 hours with sterilization.

Sophocarpine suppresses method of proliferating (plaque forming method) with virus and measures when Balb/c model mice viremia

BIAO and BEN dilution factor (COXA 9Serum during viremia)
								The plaque number	10 -2	10 -3	10 -4	10 -5	10 -6	10 -7	The cell contrast
The treatment group	Merge	168	28	12	0	0	0
								COXA 9The virus group	Merge	Merge	456	124	24	0	0

Sophocarpine is to COXA shown in the last table ₉The inhibited proliferation of virus is 90.4%～94%.

Two .COXA ₉The making of viral myocarditis and encephalitis body inner model.

The Balb/c male mice in 50 of 3～4 all ages, must be provided by triumphant laboratory animal company limited by the Sino-British joint western pul in Shanghai.Intraperitoneal injection COXA ₉Virus 1000TCID ₅₀0.1ml, cause myocarditis and meningitis model.

Experiment divides three groups: normal control group (N=10), virus control group (N=10), COXA ₉Viral infection+drug group (N=30), respectively 10 of 10mg, 20mg, 30mg with 0.1%, 0.2% and 0.3% sophocarpine 0.01ml+ normal saline 0.1ml, divide secondary to make tail vein injection respectively every day.Infect back the 8th day the aseptic eye socket of survival mice is got blood (survey neutralizing antibody), draw neck to cause death then to core dirty and brain is done the experiment of two aspects.

(1) detection of neutralizing antibody; On the Hela cell, survey three various dose sophocarpine (10mg, 20mg, 30mg) with the cytopathy political reform and survey COXA ₉The neutralizing antibody of virus.

Survey COXA with the Hela cell ₉The neutralizing antibody of virus (blood was little clear in the 8th day)

Serum dilution

1∶2

1∶4

1∶8

1∶16

1∶32

1∶64

1∶128

1∶256

1∶512

The cell contrast

Virus control

10mg

+

+

+

+

-

+

20mg

±

+

+

+

-

+

30mg

+

+

+

?-

+

COXA ₉The anti-testing result 10mg of virus neutralization was 1: 64 positive on the 8th day, and 20mg is 1: 128 positive, and 30mg is 1: 128 positive, relevant with used sophocarpine dosage (occur neutralizing antibody in the serum, the making that pathological model is described is successful).

(2) use TCID ₅₀Experiment survey various dose sophocarpine (10mg, 20mg, 30mg) on the Hela cell to COXA ₉TCID in the homogenate of viral infection myocardial cell ₅₀Observe sophocarpine to COXA ₉The inhibitory action of virus.

Survey COXA with the Hela cell ₉TCID in the homogenate of viral infection myocardial cell ₅₀

Hela cytopathy hole count/total cell hole count

BIAO and BEN dilution factor (cardiac muscle)

Sophocarpine treatment group mg/kg

10 -2

10 -3

10 -4

10 -5

10 -6

10 -7

10 -8

10mg

4/4

2/4

1/4

0/4

0/4

0/4

0/4

20mg

4/4

2/4

1/4

0/4

0/4

0/4

0/4

30mg

4/4

2/4

0/4

0/4

0/4

0/4

0/4

COXA 9Virus

4/4

4/4

4/4

4/4

2/4

0/4

0/4

The normal control group

0/4

0/4

0/4

0/4

0/4

0/4

0/4

Sophocarpine is to COXA ₉Virus titer is 10 in the viral myocarditis homogenate ^-3, virus titer is 10 in the virus control group core even slurry ^-5

Treatment group TCID ₅₀Than high 2 dilution factors of matched group, the prompting sophocarpine has antagonism COXA ₉The effect of virus.Sophocarpine on the Hela cell culture to COXA ₉The suppression ratio of virus is 99%.

(3) COXA ₉The 8th day model of the Balb/c mice of viral infection, the pathological section of heart and brain.The BIAO and BEN of heart and cerebral tissue is with 10% formalin fixed, paraffin embedding, and 10 of serial section, heart tissue and pathological changes such as cerebral tissue leukocyte infiltration, necrocytosis and degeneration are observed in HE dyeing, contrast COXA ₉The result of infection group and viral infection group and sophocarpine treatment group.

(1) COXA ₉The pathological change of viral infection Balb/c mouse cardiac muscle cell sophocarpine treatment group and infected group (attaching photo one, two, three).

Result: heart section COXA ₉Balb/c mice viral myocarditis is seen large stretch of cardiac muscle of visceral pericardium and myocardium inflammatory disorders (as shown in Figure 1) due to the virus.Sophocarpine 10mg/kg treatment group visceral pericardium cardiac muscle inflammation obviously alleviates the rarely seen a small amount of inflammatory cell of myocardium (as shown in Figure 2).Sophocarpine 20mg/kg treatment group visceral pericardium cardiac muscle and myocardium are not seen inflammation (as shown in Figure 3).

(2) COXA ₉Virus Balb/c mouse brain is organized sophocarpine treatment group and infected group pathological change (seeing Fig. 4,5,6).

Brain tissue slice, COXA ₉Balb/c mice viral meningitis due to the virus, the virus group is shown obvious ependymitis (as shown in Figure 4).Sophocarpine 10mg/kg treatment group shows that the ependymitis pathological changes alleviates (as shown in Figure 5).Sophocarpine 20mg/kg treatment group is not seen ependymitis (as shown in Figure 6).

This shows that sophocarpine is to COXA ₉Virus has tangible antivirus action, in order to treatment COXA ₉The myocarditis and the meningitis of virus animal model are effective.

Claims (4)

1. by the application in the medicine of COxsackie A9 disease that virus causes, wherein, described disease is myocarditis or meningitis in the preparation treatment for sophocarpine or its officinal salt, and the structural formula of described sophocarpine is (I)

2. sophocarpine as claimed in claim 1 or its officinal salt are treated by the application in the medicine of COxsackie A9 disease that virus causes in preparation; Wherein, the officinal salt of described sophocarpine is bromate or the hydrochlorate of sophocarpine or the fluorate of sophocarpine of sophocarpine.

3. by the application in the medicine of COxsackie A9 disease that virus causes, wherein, described pharmaceutical dosage form is any dosage form of medically approving in the preparation treatment for sophocarpine as claimed in claim 1 or its officinal salt.

4. by the application in the medicine of COxsackie A9 disease that virus causes, wherein, described pharmaceutical dosage form is powder or injection or capsule or tablet or oral liquid in the preparation treatment for sophocarpine as claimed in claim 3 or its officinal salt.

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