CN102349898B - Application of sophocarpine to preparation of medicament for treating cardiovascular diseases caused by coxsackie virus B - Google Patents
Application of sophocarpine to preparation of medicament for treating cardiovascular diseases caused by coxsackie virus B Download PDFInfo
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Abstract
The invention discloses application of sophocarpine or pharmaceutical salts thereof to preparation of medicament for treating cardiovascular diseases caused by coxsackie virus B. The sophocarpine or pharmaceutical salts thereof can be used as medicaments for treating dilated cardiomyopathy, viral endocarditis, viral valvulitis, viral pericarditis, viral coronary arteritis and hypertrophic cardiomyopathy caused by the coxsackie virus B. According to the sophocarpine or pharmaceutical salts thereof, the cure rate can be improved, the death rate can be reduced; and the sophocarpine or pharmaceutical salts thereof have no obvious toxic effects of cells, simpleness and convenience for preparation and low cost and are suitable for industrial production; meanwhile, medical resources of the sophocarpine are abundant.
Description
The application is that application number is to divide an application for " application of sophocarpine in the medicine of preparation treatment diseases induced by coxsackie B virus " for " on October 26th, 2007 ", denomination of invention " 200710047507.2 ", the applying date.
Technical field
The invention belongs to pharmaceutical field, relate to a kind of medicinal preparation, is a kind of application of sophocarpine, the particularly sophocarpine application in the medicine of preparation treatment diseases induced by coxsackie B virus specifically.
Background technology
Sophocarpine (sophocarpine) is the effective monomer that extracts in dry root from cassia leguminous plant Radix Sophorae Flavescentis and cassia leguminous plant Herba Sophorae alopecuroidis and above ground portion thereof, and molecular formula is C
15H
22ON
2, fusing point is 53-54 ℃, its structural formula is (I):
Be disclosed in one of the applicant in granted patent 99119850.6 about sophocarpine antagonism Coxsackie B virus (coxsackie virus B) and medicine preparation thereof.In addition, the present inventor has also found sophocarpine to the therapeutic effect in the disease of anti-coronavirus (SARS Virus) initiation, and the correlation technique content is in being disclosed in granted patent 03151269.4.The present inventor finds that also sophocarpine can resist echovirus (Echo virus), adenovirus (Adeno virus) and influenza virus (Influenza virus).It can be to RNA virus resisting, again can be to resisting DNA virus.Therefore, it is a kind of broad-spectrum antiviral medicament.The correlation technique content has been disclosed in another patent application 200410089431.6 of the applicant.
Domestic disease of viral infection has increasing trend.Can cause different clinical manifestations in disease of viral infection, the virus of dissimilar intestinal infection and non-intestinal group can produce clinical syndrome of the same race.On the other hand, the enterovirus of same type is under different condition, or homophyletic can not cause different clinical syndromes.The inventor finds that Coxsackie B virus (COX-Virus) can invade various organ, causes the outstanding pathological changes of this organ.as invade heart, can cause Coxsackie B virus heart disease (coxsackie Virus heart disease), invade respiratory tract and can cause upper respiratory tract infection and pneumonia (coxsackie Virus pneumonitis), invade brain and can cause aseptic meningitis and encephalitis (coxsackie Virus meningitis and encephaltides), invade liver and can cause Coxsackie B virus hepatitis (coxsackie Virus hepititis), invade pancreas and can cause pancreatitis (coxsackie Virus pancretitis), infringement skeletal muscle can cause Bornholm disease (epidemical myodynia) but can independently there be also cross occurrence in above each organ disease.The present invention is by adopting the molecular biological many rapid enzyme chain reactions (Polymerase Chain Reaction-PCR) of high-tech technological means, Coxsackie B virus ribonucleic acid (coxsackie B Virus ribonucleic acid CBV-RNA) detected from patient's blood plasma or peripheral leukocytes, can make accurately viral diagnosis.The pathological changes serious symptom person that Coxsackie B virus causes causes serious consequence even to die suddenly, and the normal persistent infection delay that has is continuous, so that organ function goes down or loses.In present existing antiviral drugs, still lack the effectively active drug of antagonism CBV.
Treat at present the disease that Coxsackie B virus infects, the clinical symptomatic treatment of all taking.In recent years once there was the scholar to report interferon and infected CVB at experiment mice
3The time or infect after 24 hours and to use the propagation of failing to block Coxsackie B virus, ribavirin (ribavirin) is the medicine mainly for adenovirus (genus DNA viruses).Though someone thinks that the RNA that can stop virus is synthetic.But effective treatment concentration of this medicine is toxic to human body.The acute CVB of the treatment such as immunosuppressant and ciclosporin mice infects without effective report, but has report can make instead with this Drug therapy that sb.'s illness took a turn for the worse.Therefore the interferon of more than addressing, ribavirin, immunosuppressant and ciclosporin are not the medicines of effective anti-Coxsackie B virus.
For seeking the active drug of antagonism Coxsackie B virus.The inventor only filters out Radix Sophorae Flavescentis simply by experiment from numerous Chinese medicine, its low toxicity and the effect of obvious anti-Coxsackie B virus is arranged.Radix Sophorae Flavescentis contains this monomer Herba Sophorae alopecuroidis of sophocarpine monomer (Sophocarpine-SC) that resists Coxsackie B virus also to be had, and high being easy to of content extracted, now made the sophocarpine injection of purity＞98%, relevant this technology contents has been open in the patent of CN99119850 at the inventor's application number.
Radix Sophorae Flavescentis is one of time-honored conventional medicament of China, begins to be stated from China's medical literature Shennong's Herbal the earliest, classifies middle product as, has the function of heat clearing and damp drying, wind dispelling insecticide, cures mainly trusted subordinate's feeling of fullness flatulence, lump in the abdomen, jaundice etc.China's pharmacopeia version in 1997 and some areas medicine specimen " Sophora alopecuroides stream soaks sheet " recorded cure mainly bacillary dysentery and enteritis (" Chinese herbal medicine (middle volume) ", in August, 1976 front page, 494～495).Zoopery in recent years proves, inhibition, analgesia and the cooling effect of Radix Sophorae Flavescentis alkaloid to nervus centralis can improve immunity of organisms, and antitumaous effect is arranged.The Radix Sophorae Flavescentis extract Radix Sophorae Flavescentis alkaloid can be treated hepatitis B, the effect that is improved clinical symptoms and liver function, some patients were HbeAg is turned out cloudy.Having document record Radix Sophorae Flavescentis alkaloid to have relievings asthma and the antiarrhythmic effect.
Summary of the invention:
The object of the present invention is to provide the application of a kind of sophocarpine in the medicine of preparation treatment diseases induced by coxsackie B virus, described sophocarpine or its officinal salt will solve the not good technical problem of medication effect of the viral dilated cardiomyopathy that in prior art, treatment is caused by Coxsackie B virus, viral hypertrophic cardiomyopathy, viral coronaritis, viral pericarditis, viral endocarditis, viral valvulitis, pneumonia, bronchitis, hepatitis, encephalitis, meningitis, pancreatitis, diabetes, Bornholm disease, epidemic pleurodynia.
The object of the present invention is to provide the application in the medicine of the Bornholm disease that preparation treatment Coxsackie B virus causes of a kind of sophocarpine or its officinal salt, wherein, the structural formula of described sophocarpine is (I):
Further, described Bornholm disease comprises epidemic pleurodynia.
The present invention also aims to provide a kind of sophocarpine or the application of its officinal salt in the medicine of the encephalitis that preparation treatment Coxsackie B virus causes, wherein, the structural formula of described sophocarpine is (I):
The present invention also aims to provide a kind of sophocarpine or the application of its officinal salt in the meningitic medicine that preparation treatment Coxsackie B virus causes, wherein, the structural formula of described sophocarpine is (I):
The present invention also aims to provide a kind of sophocarpine or the application of its officinal salt in the pancreatitic medicine that preparation treatment Coxsackie B virus causes, wherein, the structural formula of described sophocarpine is (I):
The present invention also aims to provide a kind of sophocarpine or the application of its officinal salt in the medicine of the hepatitis that preparation treatment Coxsackie B virus causes, wherein, the structural formula of described sophocarpine is (I):
The present invention also aims to provide a kind of sophocarpine or the application of its officinal salt in the medicine of the viral pneumonia that preparation treatment Coxsackie B virus causes, wherein, the structural formula of described sophocarpine is (I):
The present invention also aims to provide a kind of sophocarpine or the application of its officinal salt in the medicine of the viral bronchitis that preparation treatment Coxsackie B virus causes, wherein, the structural formula of described sophocarpine is (I):
The present invention also aims to provide a kind of sophocarpine or the application of its officinal salt in the medicine of the viral dilated cardiomyopathy that preparation treatment Coxsackie B virus causes, wherein, the structural formula of described sophocarpine is (I):
The present invention also aims to provide a kind of sophocarpine or the application of its officinal salt in the viral endocarditic medicine that preparation treatment Coxsackie B virus causes, wherein, the structural formula of described sophocarpine is (I):
The present invention also aims to provide a kind of sophocarpine or the application of its officinal salt in the viral valvulitis medicine that preparation treatment Coxsackie B virus causes, wherein, the structural formula of described sophocarpine is (I):
The present invention also aims to provide a kind of sophocarpine or the application of its officinal salt in the medicine of the viral pericarditis that preparation treatment Coxsackie B virus causes, wherein, the structural formula of described sophocarpine is (I):
The present invention also aims to provide a kind of sophocarpine or the application of its officinal salt in the medicine of the viral coronaritis that preparation treatment Coxsackie B virus causes, wherein, the structural formula of described sophocarpine is (I):
The present invention also aims to provide a kind of sophocarpine or the application of its officinal salt in the medicine of the viral hypertrophic cardiomyopathy that preparation treatment Coxsackie B virus causes, 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 pharmaceutical dosage form of the officinal salt of described sophocarpine is any dosage form of medically approving.
Concrete, described pharmaceutical dosage form is powder or injection or capsule or tablet or oral liquid.
Oral formulations and intravenous administration compare at Pharmacokinetics and bioavailability, and the physiological disposition of two kinds of preparations all meets two-compartment model.The haemoconcentration Cmax of oral liquid is lower than intravenous fluid, peak time Tmax also extends relatively, its parameter T1/2 β, K10, cLs, Auc etc. show that without significant difference (P＞0.05) two kinds of dosage forms of oral formulations and intravenous formulations have bioequivalence, and F is 80-85%.
The preparation of the medicine of sophocarpine of the present invention or its officinal salt can be with reference to the method for preparation of drug of routine, and after the dosage of having known required medicine, the preparation of medicine is that the art personnel are known.But the granted patent 99119850.6 before a kind of preparation method REFERENCE TO RELATED people of the medicine of sophocarpine or its officinal salt.
The present invention compares with prior art, and its effect is actively with obvious.Sophocarpine of the present invention or its officinal salt can effectively stop the biosynthesis of Coxsackie B virus, and enter performance antivirus action in cell, therefore sophocarpine of the present invention or its officinal salt can be treated viral dilated cardiomyopathy, viral hypertrophic cardiomyopathy, viral coronaritis, viral pericarditis, viral endocarditis, viral valvulitis, pneumonia, hepatitis, encephalitis, meningitis, pancreatitis, diabetes, Bornholm disease, the epidemic pleurodynia that is caused by Coxsackie B virus.Use sophocarpine or its officinal salt as the medicine of each relevant internal organs due to Coxsackie B virus, can improve cure rate, reduce mortality rate, and there is no obvious cytotoxic effect.And the medicine source of sophocarpine is abundant, and prepares easyly, with low cost, is applicable to suitability for industrialized production.
Description of drawings:
Fig. 1 is breast muscle's section of normal Balb/c mice, and photo shows that the chest muscle of mice is normal, has no pathological changes.
Fig. 2 is the tissue pathological slice after Coxsackie B virus infects Balb/c mice chest muscle, has shown in photo that the chest muscle transverse section section of mice has inflammatory cell infiltration, inflammatory cell infiltration around small artery.
Fig. 3 adopts sophocarpine of the present invention or the treatment of its officinal salt to be infected breast muscle's pathological section of Balb/c mice by Coxsackie B virus, after sophocarpine or the treatment of its officinal salt, and the calcification of kitchen range sexual cell.
Fig. 4 is the brain tissue slice of normal Balb/c mice, and photo shows that the cerebral tissue of mice has no obvious pathological changes.
Fig. 5 Coxsackie B virus infects the histopathology section of Balb/c mice, and photo shows the cerebral tissue focal necrosis of mice, inflammatory cell infiltration.
Fig. 6 adopts sophocarpine of the present invention or the treatment of its officinal salt to be infected the histopathology section of Balb/c mice by Coxsackie B virus, and photo has shown glial nodule under the nearly ependyma of the cerebral tissue of mice, has no obvious inflammation.
Fig. 7 is the pancreas section of normal Balb/c mice, and photo shows the pancreatic cell NIP of mice, and islets of langerhans is complete.
Fig. 8 is the pancreas pathological section that Coxsackie B virus infects the Balb/c mice, and photo shows that the pancreas of mice has obvious inflammation, and pancreatic cell destroys, and islets of langerhans destroys.
Fig. 9 adopts sophocarpine of the present invention or the treatment of its officinal salt to be infected the pancreas pathological section of Balb/c mice by Coxsackie B virus, and it is normal that photo has shown that the pancreatic cell of mice approaches, and it is complete that islet cells keeps.
Figure 10 is the liver section of normal Balb/c mice, and photo shows that the liver of mice is without pathological changes.
Figure 11 is the hepatic pathology section that Coxsackie B virus infects the Balb/c mice, and photo shows the swelling of liver cell of mice, and the endochylema endoparticle disappears, luminescence transparent, is the balloon sample and becomes.
Figure 12 adopts sophocarpine of the present invention or its officinal salt treatment Coxsackie B virus to infect the hepatic pathology section of Balb/c mice, and photo shows the rarely seen after birth inflammation of liver of mice, and pathological changes is lighter.
Figure 13 is the heart section of normal Balb/c mice, and the Epicardium of the nearly pericardium of heart of photo demonstration mice is normal.
Figure 14 is the pericarditis heart pathological section that Coxsackie B virus infects the Balb/c mice, and under the visceral pericardium of the nearly pericardium of heart of photo demonstration mice, myocardium inflammatory changes.
Figure 15 adopts sophocarpine of the present invention or its officinal salt treatment Coxsackie B virus to infect the pericarditis heart pathological section of Balb/c mice, and photo shows that the Epicardium inflammatory variation of the nearly pericardium of heart of comparing mice with the virus group obviously alleviates.
Figure 16 is the heart section of normal Balb/c mice, and the heart endocardium of photo demonstration mice and valve are without pathological changes.
Figure 17 is endocarditis and the Valvulitis heart pathological section that Coxsackie B virus infects the Balb/c mice, and photo shows endocardium and the valve inflammatory cell infiltration of mice.
Figure 18 adopts sophocarpine of the present invention or its officinal salt treatment Coxsackie B virus to infect endocarditis and the Valvulitis heart pathological section of Balb/c mice, compares with the virus group, and heart endocardium and the valve inflammatory lesion of mice alleviate.
Figure 19 is the heart section of normal Balb/c mice, and photo shows that the cardiovascular of mice is without pathological changes.
Figure 20 is the coronaritis heart pathological section that Coxsackie B virus infects the Balb/c mice, and photo shows that mice has myocardium polyangitis disease, and the wall smooth muscle cell is loose, hypertrophy.
Figure 21 adopts sophocarpine of the present invention or its officinal salt treatment Coxsackie B virus to infect the coronaritis heart pathological section of Balb/c mice, and photo shows the little blood vessel endothelium hypertrophy of myocardium of mice, compares with the virus control group, and pathological changes is lighter.
Figure 22 is the pathological examination photo of coronaritis patient cardiac muscle biopsy section, photo is (from a male, 39 years old, suddenly angina pectoris, the clinical manifestation myocardial infarction, electrocardio diagram ST section is raised, myocardium enzyme increases) show that the visible blood capillary of patient thickens, smooth muscle cell proliferation, a matter inflammatory cell infiltration.
Figure 23 is the etiological examination photo of coronaritis patient cardiac muscle biopsy section, with Figure 22 be same patient, section take off cured after, detect with In situ reverse transcription polymerase chain reaction, find Coxsackie B virus ribonucleic acid (CVB-RNA).
Figure 24 is coronaritis patient cardiac muscle biopsy section, and photo (from a male, 23 years old, angina pectoris suddenly.The clinical manifestation myocardial infarction, electrocardio diagram ST section is raised, myocardium enzyme increases.) show that patient's blood capillary thickens, smooth muscle cell proliferation, a matter inflammatory cell infiltration, and see that another blood capillary tube chamber almost completely stops up.
Figure 25 is healthy normal person's myocardium pathological section, photo (from a male, 19 years old, because of traffic accident till death.) show that blood capillary is normal.
Figure 26 Coxsackie B virus infects myocarditis, the myocardium pathological section of Balb/c mice, and photo shows that myocardial cell melts necrosis, interstitial edema, inflammatory cell infiltration.
Figure 27 adopts sophocarpine of the present invention or the treatment of its officinal salt to be infected myocarditis, the myocardium pathological section of Balb/c mice by Coxsackie B virus, and photo shows that the myocardial cell of mice shows no obvious abnormalities.
Figure 28 is the lung tissue section of normal Balb/c mice, and photo shows the thin NIP of alveolar wall of mice.
Figure 29 is the pathologic section that Coxsackie B virus infects the Balb/c mice, and photo shows that alveolar wall and the compartment of mice thicken, congestion and edema.
Figure 30 adopts sophocarpine of the present invention or its officinal salt treatment Coxsackie B virus to infect the pathologic section of Balb/c mice, and photo shows to be compared with the virus group, and the lung tissue of mice has clear improvement.
Figure 31 has shown the injection of sophocarpine or its officinal salt to the low not impact of (LVEDP) of perfusion isolated rat heart left chamber diastole, before each dosage of LVEDP and medicine than all ↑ P＜0.01,7.5 * 10
-6Gm/ml, 3.75 * 10
-5Gm/ml, after 15 minutes, LVEDP all descends, and 35 ' beginning～40 ' compare with matched group, LVEDP all descends, P＜0.05.
The specific embodiment:
Embodiment 1 in vitro tests
Cytopathic-effect inhibition assay is measured the toxicity of sophocarpine on the vero cell and the effect of antiviral (CVB3m), the result of sophocarpine and " positive drug " (Moroxydine, ribavirin) relatively on the vero cell proves that sophocarpine is low toxicity and obvious antagonism CVB is arranged
3The medicine of m
One, the compound method of sophocarpine, Moroxydine, three kinds of mother solutions of ribavirin
Sophocarpine is 100mg/ml to be diluted to 1% drug level (10000 μ g/ml) without calf serum RPMI-1640 culture fluid as mother solution, and sterile cryogenic is preserved.The compound method of Moroxydine and ribavirin is the same.
(1) three kind of medicine is to the Cytotoxic mensuration of vero
Be that 1% sophocarpine injection is used without calf serum RPMI-1640 culture fluid and is diluted to variable concentrations (continuous doubling dilution to 1 from 1: 2: 1024) add respectively the hole with concentration.Each dilution factor is 4 multiple holes (and establishing normal control).Cultivated 4 days, every day, observation of cell toxicity, calculated half toxic dose (TD
50) and maximal non-toxic dosage (TD
0).The same sophocarpine of the dilution process of Moroxydine and ribavirin.
Result:
Table 1: the toxic action (1% concentration two-fold dilution) of sophocarpine injection to the Vero cell
Above result shows that concentration is that 1% sophocarpine is diluted at 1: 8 o'clock (being 312ug) when following, the toxicity of vero cell disappeared, and TD
0Be 312ug/ml, TD
50Be 624ug/ml.
Table 2: guanidine hydrochloride (ABOB) is to Vero cell toxicity test (1% concentration two-fold dilution)
Above result shows, concentration be 1% guanidine hydrochloride be diluted to 1: 32 (the 2nd day and the 4th day) when following (concentration be 78ug/ml when following toxicity to the Vero cell disappear, TD
0Be 78ug/ml, TD
50Be 156ug/ml, toxicity surpasses sophocarpine.
Table 3: ribavirin (triazole) is to Vero cell toxicity test (1% concentration two-fold dilution)
Above result shows, concentration be 1% ribavirin be diluted to 1: 64 (the 2nd day and the 4th day) when following (concentration be 39ug/ml when following toxicity to the Vero cell disappear, TD
0Be 39ug/ml, TD
50Be 78ug/ml, toxicity more surpasses sophocarpine.
(2) three kinds of medicine anti-CVB on the Vero cell
3The effect of m
Cytopathic-effect inhibition assay is measured sophocarpine injection anti-CVB on the Vero cell
3Effect (the CVB of m
3M concentration is 100TCID
50)
Test shows, sophocarpine injection at 312ug/ml so that obvious antagonism CVB all to be arranged down to 5ug/ml
3The effect of m.
Cytopathic-effect inhibition assay is surveyed ribavirin (triazole) anti-CVB on the Vero cell
3M experimental result (CVB
3M100TCID
50)
Cytopathic-effect inhibition assay is surveyed guanidine hydrochloride (ABOB) anti-CVB on the Vero cell
3The m experimental result
Above test shows, guanidine hydrochloride and ribavirin are all without obvious antagonism coxsackie B
3The m virus function.
(3) the variable concentrations sophocarpine reaches with mtt assay, crystal violet vital staining
3[H] mixes method and measures cellular energy metabolism rate, cell survival rate and DNA anabolism rate
(1) 3[H] the TdR method of mixing measures sophocarpine to the protective effect of Vero cell DNA anabolism rate: use 3[H] TdR mixes method and be the result of variations of the measured value of 5 o'clock Vero cells at the sophocarpine of variable concentrations in infection multiplicity (CVB3m-MOI:5), and is as shown in the table:
Table 4:3[H] TdR mixes method: the mensuration of variable concentrations sophocarpine to the anabolic protective effect of Vero cell DNA
MOI:5 (the viral infection plural number is 5)
Annotate: CPM is for dodging the umber of pulse of measuring on dry device at liquid
**P＜0.01
***P＜0.001
When result was 5 when the viral infection plural number, sophocarpine all had protective effect to the DNA anabolism of vero cell when 200ug/ul-3.12ug/ml, compare P＜0.01-0.001 with the virus control group.Minimum effective drug concentration (MTC) is 3.125ug/ml.
(2) protective effect of tetramethyl azo azoles salt (MTT) method mensuration sophocarpine to the Vero cellular energy:
Table 5: tetramethyl azo azoles salt (MTT) method is measured the variable concentrations sophocarpine protective effect of Vero cellular energy metabolism is measured
MOl:5 (the viral infection plural number is 5 o'clock)
Annotate: each dosage group is compared p＜0.05 for variant with the virus control group,
* *＜0.01 is unusual notable difference
Result: when sophocarpine is at 200ug/ul to 3.125mg/ml when CVB3mMOI:5 all the energy metabolism to the vero cell have protective effect to compare p＜0.05-0.001 with the virus control group.
(3) protective effect of crystal violet competent cell staining survey various dose sophocarpine to the vero cell survival rate:
Table 6: the protective effect of crystal violet competent cell Determination Staining variable concentrations sophocarpine to the vero cell survival rate
(when CVB3mMOI:5)
When the viral infection plural number was 5, sophocarpine injection concentration all had protective effect to the survival rate of vero cell when 200ug/ul to 3.125mg/ml.Its OD measured value rises with sophocarpine injection concentration, P＜0.05-P＜0.001.
In sum, the Vero cell DNA anabolism rate of sophocarpine to CVB3m, energy metabolism and cell survival rate all have protective effect.
Embodiment 2 in vivo tests
(1) by the research of sophocarpine injection to antagonism virus in the moulding Mice Body, all demonstrate its low toxicity and the effect of obvious anti-CBV arranged, can survive more than 50 days to the mice of medication treatment group.And the matched group of medication 100% was not died from 8 days, illustrated that sophocarpine can reduce the mortality rate of infecting mouse, extended life cycle, and each cells of organs of virus group is downright bad companion's inflammatory cell infiltration obviously.And the heart for the treatment of group, lungs, liver, pancreas, brain, skeletal muscle are all without significantly pathological change, therefore sophocarpine antagonism CVB
3The effect of m, thus the disease of each internal organs due to coxsackie B there is obvious therapeutic action.
(2) with cytopathy political reform (cell pathogenic effect-CPE)
Be determined at the effect of sophocarpine anti-CVB3m in the Balb/c mice viremia phase serum that CVB3m infects on the Vero cell with cytopathy political reform (micromethod), relatively the TCID for the treatment of group and virus group
50(Tissue culture infectious dose) calculates sophocarpine to the suppression ratio (%) of CVB3m
The effect of microdetermination sophocarpine (mice viremia) anti-CVB3m in stopping
Upper table shows, treatment group serum-virus titre is 10 as calculated
-3TCID
50, the virus control group is 10
-5TCID
50, press Read Muench formula and calculate sophocarpine in vivo to CVB
3The suppression ratio of m is 99%.
Embodiment 3 Bornholm diseases and chest pain
Epidemic pleurodynia: 5 types of Coxsackie B virus all can cause myalgia and myasthenia.Be 2～5 days incubation period, can extend to for 2 weeks.Main manifestations is heating and paroxysmal myalgia, and myalgia can be involved whole body skeletal muscle.And take the breast abdominal part as common, especially diaphram is the most easily got involved, and is normal and the aseptic meningitis while is concurrent.The chest pain weight differs, and severe one even causes shock.When chest muscle is movable, chest pain is more acute.Chest X-ray is without abnormal discovery.At present treatment is with analgesic or with morphine and codeine.After the analgesic treatment, a couple of days can be alleviated.But outbreak repeatedly can be arranged later on, and major lesions is in inflammation and the necrosis of Skeletal Muscle Cell.Record patient's blood WBC and have CVB-RNA to exist can to make a definite diagnosis, adopt the medicine of effective anti-CVB virus such as sophocarpine injection in time to resist CVB virus if in time use PCR, the state of an illness is controlled.The inventor confirms in the test of Wistan pharmacokinetics in rats that at sophocarpine the concentration of sophocarpine in skeletal muscle is considerably beyond blood concentration.When blood drug level was 7.335ug/ml, the concentration in skeletal muscle was 22.632ug/ml; When blood drug level was 4.7870ug/ml, the concentration in skeletal muscle was 23.148ug/ml; When blood drug level was 2.8479ug/ml, the concentration in skeletal muscle was 7.219ug/ml; When blood drug level was 1.222ug/ml, the concentration in skeletal muscle was 3.6587ug/ml; When blood drug level was 0.4127ug/ml, the concentration in skeletal muscle still had 1.4262ug/ml.Illustrate that the application of sophocarpine in COxsackie myositis (epidemic pleurodynia) is definite.
As Fig. 1, Fig. 2, shown in Figure 3, the inventor establishes normal group, virus control group and sophocarpine treatment group by the animal model that CVB infects the Balb/c mice, take off neck after 10 days lethal, get its chest muscle check pathological section, the chest muscle pathological changes that confirms the sophocarpine treatment obviously alleviates, close to normal group, illustrate that sophocarpine is effective in the striped muscle inflammation that treatment CVB causes.
Embodiment 4 Coxsackie B virus (aseptic) encephalitis and meningitis
Aseptic meningitis is a kind of syndrome very common in enterovirus infection (Coxsackie B virus especially.Be difficult for when popular in summer differentiating mutually with light-duty epidemic encephalitis type B.Characteristics of incidence is heating, and similar cold symptoms headache, pharyngalgia in succession occur or feels sick, vomits, and muscular soreness or weak is arranged sometimes.Leukocytosis in stiffness of nape and back, cerebrospinal fluid (CSF) appears in serious symptom person.Generally in 100～200 left and right, occasionally can reach more than 1000.Shanghai is year follow-up period in nineteen sixty～1970, once is separated to COXA from aseptic meningitis or paralysis disease
9, COXB
1, B
3, B
5With Echo virus
3,4,6,9,13,16Etc. type.Be separated to COXA in 1964
9And COXB
4Virus is many.Many systems are isolated in cerebrospinal fluid (CSF).Rise with neutralizing antibody after being ill.Now use high-tech polymerase chain reaction,PCR (Polymerase Chain Reaction PCR) technology to detect above-mentioned every virus rapidly from CSF or periphery WBC and make a definite diagnosis, can in time adopt sophocarpine injection to treat.Zoopery in recent years proves, Radix Sophorae Flavescentis alkaloid has inhibition, analgesia and cooling effect to nervus centralis.The inventor in Wistar pharmacokinetics in rats test, proves that sophocarpine distribution half-life in the rat body is 32.6583min at sophocarpine, and the removing half-life is 143.4min, and the sophocarpine concentration in cerebral tissue is far away higher than blood concentration and hold time longer.When blood drug level was 7.335ug/ml, the concentration in cerebral tissue was 40.116ug/m; When blood drug level was 4.7870ug/ml, the concentration in cerebral tissue was 21.762ug/ml; When blood drug level was 2.8479ug/ml, the concentration in cerebral tissue was 11.208ug/ml; When blood drug level was 1.222ug/ml, the concentration in cerebral tissue was 5.4788ug/ml.And can measure at 9～15 hours.Sophocarpine of the present invention or its officinal salt can see through the brain barrier, are conducive in encephalitis, meningitis to the CoxsachieB Removing, pathological changes to be recovered.
As Fig. 4, Fig. 5, shown in Figure 6, the inventor establishes normal group, virus control group and sophocarpine treatment group by the animal model that CVB infects the Balb/c mice, take off neck lethal after, get its cerebral tissue chest muscle check pathological section, glial nodule under the nearly ependyma of cerebral tissue of confirmation sophocarpine treatment, have no obvious inflammation, close to normal group, illustrate that sophocarpine is effective in the meningitis inflammation that treatment CVB causes.
Embodiment 5 Coxsackie B virus pancreatitis and diabetes
Pancreatitis: except biliary tract disease, heavy drinking, gluttony and Salmonella or streptosepticemia can cause beyond acute pancreatitis, also can cause acute pancreatitis after the infection such as the infectious disease of acute viral infection such as acute parotitis, infectious monocytosis, Coxsackie B virus, Echo virus.Acute pancreatitis is to cause the chemical inflammation of pancreatic tissue autodigestion after being activated in pancreas by pancreatin.Clinically with acute epigastrium pain, feel sick, characteristics that vomiting, heating, blood and amylase in urine increase.The lighter is take the pancreas edema as main.The state of an illness has self limiting sometimes, can recover fully after a few days.Minority is in a bad way, and pancreatic hemorrhage is downright bad, and companion's peritonitis because digestive enzyme and slough liquid can be transported to whole body by blood circulation and Lymphatic channel, causes many organ injuries, becomes the multiple complications of acute pancreas, and the shock death rate is high.What have is converted into chronic pancreatitis, and the complete and occurrence of diabetes of endocrine is wherein arranged more than 50%.
General concept thinks that the morbidity of diabetes is relevant with heredity, according to domestic report diabetics, positive family members Shi Zhezhan 8.7% is arranged, and is reported to 25%～50% abroad.But not yet bright loud and clear about the mode of heredity, the polygenic inheritance defective may be arranged.Problem be except 25%～50% diabetes be foreign genetic element, the factor of viral infection can not except, COXB virus especially.Certain areas significantly raise at the sickness rate of the popular rear diabetes of viral infection, and the anti-coxsackie B of patients serum
4Virus titer raises, and illustrates that Coxsackie B virus infects directly or indirectly (by hideing from the body immunity) damage islet tissue (particularly young diabetics).In high-tech PCR detection patient blood WBC, CVB-RNA existed and can make a definite diagnosis rapidly as immediately adopting at that time in morbidity.Use anti-coxsackie B active drug sophocarpine treatment.
As Fig. 7, Fig. 8 and shown in Figure 9, the inventor has obvious inflammation by the pancreas of finding viral group in Balb/c animal model due to COXB virus, and pancreatic cell destroys, islets of langerhans destroys, and sophocarpine treatment group pancreatic cell approaches normally, and it is complete that islet cells keeps.
Embodiment 6 Coxsackie B virus hepatitis
Can ill virus hepatitis (coxsackie virus B heptitis) after Coxsackie B virus infects, onset is mostly anxious, and fear of cold, heating, outstanding symptom are arranged is inappetence, feel sick, vomiting, epigastric discomfort, abdominal distention are weak etc.Onset can have obvious myalgia intend like rheumatic fever or headache, upper respiratory tract infection symptoms are arranged, and just like influenza, minority case stomachache is very acute, can be mistaken for acute abdomen.The common liver enlargement of clinical manifestation, tenderness, lab testing have glutamate pyruvate transaminase (SGPT), glutamic oxaloacetic transaminase, GOT (SGOT), alkali phosphatase (AKP), γ-paddy acyl transpeptidase, and (γ-GT) etc. increases.More than the visible swelling of liver cell of hepatic pathology cut sections for microscopic examination (increasing 3 times than normal liver), the endochylema endoparticle disappears mostly, shinny therefore transparent (claiming the balloon sample to change), the hepatocyte spotty necrosis, when serious, hepatocyte dwindles, core loses, becoming the red circular corpusculum that dyes is the eosinophilic body, and the hepatic cords arrangement disorder makes the hepatocyte trabecularism unclear.Can survey with molecular biology state-of-the-art technology polymerase chain reaction,PCR (Polymerase Chain Reaction PCR) and have Coxsackie B virus ribonucleic acid (CVB-RNA) in patient's blood leukocytes (WBC) and make a definite diagnosis rapidly, and increase with neutralizing antibody subsequently, to the COXB viral hepatitis if in time adopt sophocarpine injection treatment ought to cure or anti-sick to delay property and chronic development here.
The inventor in the test of Wistar pharmacokinetics in rats, confirms that the concentration in liver (liver) tissue surpasses blood drug level at sophocarpine.When blood drug level was 4.787ug/ml, the concentration in liver organization was 5.732ug/m; When blood drug level was 2.8479ug/ml, the concentration in liver organization was 5.3758ug/ml; When blood drug level was 1.222ug/ml, the concentration in liver organization was 4.1692ug/ml; When blood drug level was 0.4127ug/ml, the concentration in liver organization was 3.3413ug/ml.Such pharmacokinetics basis is useful to the CVB virus of removing in liver.
As Figure 10, Figure 11, shown in Figure 12, the inventor establishes normal group, virus group and treatment group in the Balb/c model that infects CBV, getting its liver does in the contrast of pathology cut sections for microscopic examination, the hepatic lesions of finding the sophocarpine treatment group obviously alleviates, close to normally, illustrate that viral hepatitis is favourable due to sophocarpine treatment CBV.
Embodiment 7 acute viral pericarditises
Acute pericarditis is the inflammation of pericardium viscerale and parietal layer, and except rheumatic fever, tuberculosis and antibacterial infected, the sickness rate showed increased (Coxsackie B virus is one of Etiological) that infects of poison due to illness in recent years can be divided into cellulose protein and exudative two kinds.Take male, person between twenty and fifty as common, premorbid often has the upper respiratory tract infection history several weeks, and onset is hurried, shows as violent chest pain, heating, and majority can be heard pericardial friction rub, and liquid oozes out rear friction sound and disappears.Coxsackie B virus occasioner is take exudative as many.Pericardial effusion in various degree can be arranged in pericardium, mostly be the liquid of Huang Erqing, some can spontaneous recovery, if but body fluid accumulate in a large number within a short period of time, the tamponade of cardiac trigger.During acute pericarditis, Epicardium has inflammatory variation in various degree, if scope more extensively can be described as perimyocarditis or myopericarditis.Inflammation also can be involved mediastinum and pleura.With PCR method can find CVB-RNA can make a definite diagnosis from peripheral leukocytes or in paracentesis pericardii liquid this moment.Increase rapidly as liquid, pericardium can't stretch to adapt to the variation of its capacity, make the hurried rising of intrapericardial pressure power, get final product the cardiac trigger pressurized, causing ventricular diastole to fill is obstructed, and make peripheral venous pressure rising, blood pressure drops, the pulse pressure obvious tachycardia of companion that diminishes, when serious, heart stroke reduces, and shock can occur.Generally adopt clinically heteropathy and hormone therapy.Hormone therapy is effective to suppressing oozing out of pericardial effusion, but is etiological treatment, and hormone uses the breeding that instead can increase virus in acute stage.
As Figure 13, Figure 14 and shown in Figure 15, under the visceral pericardium of the inventor by the nearly pericardium of heart of the mice of discovery virus group in Balb/c animal model due to COXB virus, the cardiac muscle inflammatory changes, and after using the sophocarpine injection treatment, the Epicardium inflammatory of nearly pericardium changes and obviously alleviates.
The viral endocarditis of embodiment 8 and valvulitis
Past is thought always clinically that infective endocarditis (infectious endocarditis) is generally all thought and betides on the basis of original rheumatic valvular disease, in recent years due to virologist, pathologist and clinical scholar's research and concern, find that Coxsackie virus also can cause endocarditis and valvular heart disease, and often betided on normal heart and valve and cause valvular insufficiency and narrow.
As Figure 16, Figure 17 and shown in Figure 180, in the Balb/c animal model that the inventor's CVB infects, after viral infection, photo shows endocardium and the valve inflammatory cell infiltration of mice, after adopting the treatment of sophocarpine of the present invention or its officinal salt, the endocarditis of mice and Valvulitis pathological changes alleviate, and prove that sophocarpine or its officinal salt are effective to endocarditis and the sick treatment of valvulitis of CVB viral infection.
The heart disease of embodiment 9 coronary artery inflammation
The coronaritis of viral infection (virus infected coronary arterietis), often occur in little coronary artery (micorcoary artery), can cause a series of signs of myocardial ischemia, as coronary vasospasm or obturation, can cause angina pectoris, then pour into arrhythmia.The visible ST section of EKG changes, the R ripple reduces, pathologic Q ripple.If any ST section rise obturation will watch out for coronule shape arterial lumen and the myocardial infarction of initiation, the myocardial infarction that little coronaritis causes is different from the myocardial infarction that large coronary artery causes, because little diameter coronarius is only 50～150um, this section blood vessel can not be detected by coronarography.According to the resistance theory, coronary vasodilator can be divided into the large coronary artery of visceral pericardium and the little coronary artery of endocardium.When normal, the large coronary resistance of visceral pericardium (RL) is very little, accounts for 10% of coronary vascular resistance (R); And the little coronary artery resistance of endocardium (Rs) is very large, accounts for the 90% resistance function of R.Can reach 90～180ml/min blood when under normal circumstances, aortic pressure is 12.0kPa (90mmHg).This ratio (aortic pressure/blood flow) is called vascular resistance (ROCP/Q).Although being preced with greatly tremulous pulse also can be to blood vessel drag overall generation MC, be the little blood vessel of 10～40um but the main position of change in resistance is myocardium internal diameter, therefore think that at present it is to cause myocardium degeneration and myocardiac main cause that small coronary artery disease becomes, when myocardial damage can cause cardiac insufficiency to a certain extent.Observation according to my institute cuts into slices to 63 routine patients' endocardium biopsy reason has 11 examples to have small coronary artery disease to become, and has 7 routine patients that viral myocarditis and myocardiac medical history were arranged in 11 examples, accounts for 63.6%.In tissue slice, visible arterial wall thickens because inflammation produces, but because of the inflammation time longer, have no inflammatory cell infiltration.This 7 routine patient's pathological section is done the detection of molecular biology original position reverse-transcription polymerase chain reaction after taking off cured processing all can find to find the positive signal of COX-B-RNA in coronary arterial wall.Illustrate that it is to cause the consequence of coronary artery inflammation, pathology photo such as Figure 22, Figure 23, Figure 24 and Figure 25 due to the COXB viral infection that small coronary artery disease becomes.
Since 1975, as if existing multidigit scholar report is observed the sickness rate that respiratory tract infection has increased acute myocardial infarction, and majority is youngster, and what have only has 19 years old.Think that the heart disease performance after this viral infection may be relevant with coronaritis.Burch, Silver etc. find that in the experiment of COX viral infection Balb/c mice extensive coronary microvascular is narrow, and the blood vessel profile is unclear, and mononuclear cell and lymphocytic infiltration are arranged around local filling defect, lesion vessels and blood vessel wall.Utilizing in recent years round pcr CVB-RNA to be detected in the lymphocyte of lesion vessels and infiltration exists.
In order to confirm the effect of sophocarpine treatment Coxsackie B virus coronaritis, the inventor infects the Balb/c mice with CVB, establishes viral group, treatment group, normal group and matched group.The result demonstration, the coronary artery pathological changes improvement after the sophocarpine treatment obviously is better than the virus control group, illustrates that the viral coronaritis of sophocarpine treatment is effectively, pathology photo such as Figure 19, Figure 20 and shown in Figure 21.
In more than the 600 routine Patients with Viral Myocarditis that my institute accepts for medical treatment in recent years, there are 264 examples to be viral cardiomyopathy, the rabat postero-anterior position shows that podiod increases, in Echocardiography finding cardiac dilatation, ventricle wall plumpness or attenuation, Heart function fails, periphery WBC or can find the ribonucleic acid (CVB-RNA) of Coxsackie virus B in myocardium biological tissue with high-tech molecular biology polymerase chain reaction,PCR (Polymerase Chain Reaction-PCR).Obvious angina pectoris sample outbreak symptom often appears clinically.according to us, majority of cases is wherein made the result that myocardium live body pathological section observes with micro mirror and all show little coronary artery wall thickening in cardiac muscle, in the normal myocardium of luminal stenosis and traffic death, coronary artery is compared and is thickened very obviously (pathology photo such as Figure 22, Figure 23 and shown in Figure 25), be found to be patient's 182 examples of angina pectoris sample outbreak, before treatment, electrocardiogram shows that precordial lead ST section is obviously forced down and (descends 0.5～1.5mV), what have is inverted with the T ripple, above case is divided into two groups and treats, sophocarpine treatment group 124 examples, glucose, insulin, potassium chloride (GIK) treatment 58 examples, there is complete ecg data to compare before and after getting treatment, observe two groups of cases [NST (number that leads that the ST section descends) and ∑ ST (addition number that each ST-Segment descends)] and estimate the myocardial ischemia that sophocarpine causes coronaritis due to Coxsackie B virus, myocardial oxygen consumption and to the improvement situation of myocardial damage, see following table for details: NST before and after sophocarpine and GIK treatment, comparison mVX ± SD of ∑ ST
Relatively front with the treatment of this group
(1)P＜0.05
(2)P＜0.01
Illustrate that sophocarpine is better than the GIK group in the effect aspect NST and ∑ ST, coronary artery Myocardial Ischemia and oxygen consumption increase that prompting antagonism Coxsackie B virus causes are improved effect.
The impact of embodiment 10 sophocarpine on isolated rat heart coronary artery blood flow (CFml/min)
The preparation of isolated heart and grouping
100 of 270～400gm male SD rats are adopted in this experiment, divide each 50 of normal hemoperfusion group and hypoxia hemoperfusion group, two groups are all adopted the sophocarpine injection of 4 kinds of unified variable concentrations to carry out perfusion, to observe sophocarpine injection to the effect of heart coronary artery blood flow (CFml/min).Various dose is 0.3x10
-6Gm/ml (N=10), 1.5x10
-6Gm/ml (N=10), 7.5x10
-6Gm/ml (N=10), 3.75x10
-5Gm/ml (N=10).
Above SD rat is with 20% urethanes 5ml/kg intraperitoneal anesthesia, the heparin of intraperitoneal injection simultaneously 1u, after anesthesia, be fixed in operating-table, open rapidly breast and core dirtyly, be placed in the cold perfusion buffer that is added with heparin, heart is hung on the Langendorff perfusion system rapidly after separating aorta, perfusate is through the retrograde perfusion heart of aorta, and perfusate (HK) is with 95%O
2And 5%CO
2Saturated, and to adjust PH be 7.4, keeping perfusion pressure is 100cmH
2O, the perfusate temperature is that 37.5 ℃ of room temperatures are 22.25 ℃.Be that the 0.1mm silver wire electrode is placed in respectively right ventricle, right atrium and left atrium, recording ecg with 3 diameters.
Experimental technique:
Perfusion 15min after above end of operation after heartbeat is steady, begins experiment.
(1) normal perfusion is to sophocarpine injection group (4 groups), and in perfusate, sophocarpine injection concentration is respectively
0.3x10
-6gm/ml(N＝10)、1.5x10
-6gm/ml(N＝10)、7.5x10
-6gm/ml(N＝10)、3.75x10
-5gm/ml(N＝10)。Continue recording ecg 60min, every 5min surveys a coronary flow (CFml/min).
(2) hypoxia perfusion matched group with 37.5 ℃ without 95%O
2And 5%CO
2Saturated newly joining after perfusate perfusion 30min again with 95%O
2And 5%CO
2The saturated perfusate perfusion 30min of 37.5 ℃, in hypoxia hemoperfusion group sophocarpine liquid injection (4 groups) perfusate, sophocarpine injection concentration is respectively
0.3x10
-6gm/ml(N＝10)、1.5x10
-6gm/ml(N＝10)、7.5x10
-6gm/ml(N＝10)、3.75x10
-5gm/ml(N＝10)。
(above 95%O
2And 5%CO
2Saturated and without 95%O
2And 5%CO
2Saturated perfusate all adds the sophocarpine injection of the same concentration in both, continues recording ecg 60min, and every 5min surveys a CF)
Result
(1) normal oxygen hemoperfusion group, after rat heart exsomatizes, each hemoperfusion group CF reduces gradually, but 7.5x
10-6In gm/ml group, before 45min all higher than matched group, especially after 50～55min apparently higher than matched group P＜0.05～0.01, the speed that can delay CF decline under normal oxygen perfusion be described, see Table 7.
(2) hypoxia hemoperfusion group: after isolated rat, each hemoperfusion group CF all descends gradually, 7.5x
10-6The CF value of gm/ml group is higher than matched group, and especially when 40min (after recovering normal oxygen perfusion), its CF value is apparently higher than matched group, and P＜0.01 sees Table 8.
Illustrate that sophocarpine injection can increase the coronary flow of SD isolated rat heart and the resisting oxygen lack of increase heart.
Embodiment 11 sophocarpine are on the impact of dog at systemic heart blood flow coronarius
Sophocarpine has allevating angina pectoris, coronary artery dilator to increase the effect of coronary flow (coronary blood flow fraction-CF) in Chinese medicine.in another embodiment of the invention, the coronary artery blood flow at systemic heart to dog is studied, after experimental dog anesthesia, get right lateral position, along the 4th intercostal incision, separating muscle enters the breast chamber, expose heart, the free left circumflex artery of identification coronary artery, make the Doppler ultrasonic probe just Perfect Ring around, be coated with conductive electrode cream around probe, connect the Doppler flow measuring instrument, measure the circumflex branch of coronary artery flow, first measure each parameter after stability of flow, later on every 5～10min surveys once, then be calculated to coronary flow per minute each 10 seconds. add glucose 20ml with sophocarpine 6mg/kg, find after intravenous injection that slowly sophocarpine can increase flow coronarius (CF ml/min).Be 51.00 ± 3.46 before medication, with being 75.5 ± 10.39 after sophocarpine treatment, when waiting capacity glucose to compare as 51.00 ± 3.46.Illustrate that sophocarpine injection can increase coronary flow, compare with reaching glucose before medication, statistical significance (t=4.3852P＜0.001) is arranged.Be conducive to improve small circulation coronarius in coronary artery, heart, be conducive to the recovery of damaged myocardium tissue after viral infection.
Comparison with dog arteries and veins flow (CF) before and after sophocarpine injection
A=is with before sophocarpine, B=with sophocarpine after, C=contrasts with glucose
Conclusion: sophocarpine injection can increase coronary flow, compares with reaching glucose group before medication, and t=4.3852 p＜O.001
Embodiment 12 dilated cardiomyopathies
Dilated cardiomyopathy (Dilated cardiomyopathy DCM) principal character is that a side or the bilateral chambers of the heart enlarge, and myocardium shrinkage function goes down or exhaustion, produces congestive heart failure.Be called congestive cardiomyopathy in the past.The primary disease mortality rate is high.It is generally acknowledged that the mortality rate that symptom occurs in rear 5 years is 40%～50%.The cause of disease of primary disease it be unclear that.Except idiopathic, Familial Occurrence, think that in recent years viral infection is its main cause.The primary disease onset is slow, and is how just medical when clinical symptoms is obvious.As out of breath, even orthopnea, edema, hepatomegaly are with dissimilar arrhythmia.18 examples that the inventor once accepted for medical treatment core Ji hospital are done the survey inspection of PCR in peripheral blood WBC, and wherein 6 examples are positive and account for 33.3%, and the part myocardium cell necrosis is seen in 13 routine heart biopsy pathology sections wherein, and by fibrous tissue is replaced, cardiac cell nucleus increases.And cured rear the detection with In situ reverse transcription polymerase chain reaction taken off in section all can find the positive signal of CVB-RNA, illustrates that this group dilated cardiomyopathy is relevant with the Coxsackie B virus persistent infection.
(1) observe sophocarpine to the impact of dilated cardiomyopathy heart pathological section, do endomyocardial biopsy and contrast before and after being preferably in patient treatment, but because intracardiac flesh biopsy is traumatic inspection, unsuitable rechecking, also be difficult for being accepted by patient, therefore the inventor has developed the animal model COXB virus inoculation 270 days of viral cardiomyopathy and has caused with mankind's clinical findings dilated cardiomyopathy mutually attached, 5 of matched groups, 6 of sophocarpine treatment groups, in the time of the 256th day, sophocarpine treatment group 20mg/kgd
-1Intraperitoneal injection) be total to 14d, matched group is with the intraperitoneal injection of volume normal saline.Result still can show myocardial cell atrophy, arrangement disorder in two groups of cardiomyopathy model cardiac muscle biopsy sections, replaced by fibrous tissue, and treatment group also has no improvement, and possible pathological changes has arrived irremediable stage.But, in the homogenate of matched group cardiac muscle, RNA (pg) is 0.8058pg ± 0.3302pg, in the homogenate of sophocarpine treatment group cardiac muscle, RNA (pg) is 0.0582pg ± 0.025pg, compare with matched group, sophocarpine is 92.77% to the clearance rate of CVB-RNA, illustrate that the Balb/c mice infecting CVB in the time of the 270th day, still can find CVB-RNA in myocardium homogenate.Prompting CVB causes viral myocarditis can be persistent state.Treat the CVB-RNA concentration that still can obviously reduce in the homogenate of animal model cardiac muscle with sophocarpine.
(2) impact of sophocarpine injection on cardiac function
1. can reduce that the phase is pressed (LVEDP) to the diastole of left chamber: LV Diastolic is the pressure of phase (left ventricles end diastolic pressure) not.When experimental SD isolated rat is surveyed the experiment of coronary flow, through left atrium, the one horizontal capsule that fills with normal saline is inserted left ventricle, connect pressure transducer, the pressure of foundation in water pocket is adjusted to the variation of recording left ventricular pressure power after 5mmHg.3 diameter 0.1mm silver wire electrodes are placed in respectively right ventricle, right atrium and left ventricle place, recording ecg, can be not left chamber diastole phase pressure not, and ± impact of dp, dt. before hypoxia perfusion matched group and various dose group and perfusion, rising is arranged more all, but do not reach statistical significance, recover after normal oxygen perfusion 35min and respectively organize LVEDP and all descend, significantly lower than 0x10
-6Gm/ml, P value＜0.05.See Figure 31 and table 9.
2. on the impact of ± dp/dt, all decline after each group ± dp/dt hypoxia perfusion.Minimum during 30min, recover to rise to some extent after normal oxygen, but still lower than before the hypoxia perfusion.7.5x10
-6Mg/ml group ± dp/dt fall is respectively organized little than other, after 35min each time point ± dp/dt apparently higher than other group (p＜0.01 〉.Illustrate that sophocarpine can improve heart contraction and diastolic function.
Result: dilated cardiomyopathy has belonged to late period in this experiment, although sophocarpine fails to change its pathologic structure, to remove CVB-RNA improve LVEDP and ± helpful aspect the cardiac function such as dp/dt.
Embodiment 13 heart disease caused by viruses
heart disease caused by viruses (viral heart disease) comprises viral myocarditis (viral myocarditis), viral dilated cardiomyopathy (viral dilated cardiomyopathy), viral hypertrophic cardiomyopathy (viral hypertrophic cardiomyopathy), viral coronaritis (viral coronary arterietis), the diseases such as viral pericarditis (viral pericarditis) and viral endocarditis (viral endocartitis), take the change of coxsackie b virus occasioner as common.To Coxsackie B virus (coxsacuie B virus-CBV), there is no so far effective antagonism medicine.The inventor is by years of researches, filtered out unique low toxicity simply from numerous Chinese medicine and the Radix Sophorae Flavescentis of antagonism CVB effect is arranged, and wherein contains the effective ingredient that resists CVB-sophocarpine monomer (Sophocarpine is called for short SC).The biosynthesis (comprising the synthetic of protein) of obvious antagonism Coxsackie B virus is arranged and enter about open in the patent of the inventor's application number CN99119850 (sophocarpine alkali is as preparation Coxsackie B virus myocarditis etiological treatment and the purposes) low toxicity of this technology contents and bring into play antivirus action in cell.Prove for dynamic test at the body giving drugs into nose to the wistar rat, sophocarpine distribution half-life in the rat body is 32.6583min, the removing half-life is 143.4min, meet two-compartment model, in the concentration of brain, heart, gastrointestinal tract, lungs and liver and skeletal muscle apparently higher than blood concentration, and it is longer to hold time, be conducive to improve the clinical symptoms that infects each organ pipe disease that causes because of the COXB viral infection, and the removal that is conducive to COXB virus.And the concentration of confirmation in heart (heart) is considerably beyond blood drug level.And the time of keeping is longer.When blood drug level was 7.335ug/ml, the concentration in heart tissue was 15.622ug/ml; When blood drug level was 4.7870ug/ml, the concentration in heart tissue was 12.149ug/ml; When blood drug level was 2.8479ug/ml, the concentration in heart tissue was 6.9815ug/ml; When blood drug level was 1.222ug/ml, the concentration in heart tissue still had 3.9333ug/ml, and still can measure at 9～15 hours.Therefore the treatment to above heart disease caused by viruses is all useful.Viral myocarditis, the most perfect definition refer to a kind of heart disease of the simple idiomuscular inflammation of the heart after viral infection, are mainly caused by Coxsackie B virus.There is the precursor symptom of viral infection in 1～3 week of premorbid, as heating, asthenia, feel sick, vomiting, then visible cardiopalmus, chest pain, dyspnea, various arrhythmia, serious symptom can cause cardiogenic syncope or acute pump failure and die suddenly.The visible heart cell of endocardium biopsy pathology section melts, interstitial edema, inflammatory cell infiltration.The heart section finding of experimental Balb/c mice through treating, viral contrast are compared obviously improvement or approaching normal of virus, and pathological section is as shown in Figure 26 and 27.
Annotate: heart preparation is fixed in 100% buffered formalin liquid, and conventional dehydration, paraffin embedding 5um quick-frozen flat cutting are done HE, Masson, V-G, Path dyeing, and every section is observed in detail.
Embodiment 14 Coxsackie B virus pneumonia
Respiratory tract infection: the respiratory tract disease that virus causes accounts for the more than half of acute respiratory disease, and especially Coxsackie virus (CVB-is also one of main virus) causes pneumonia, bronchiolitis is common.In today of antibiotic medicine extensive use, most of bacterial respiratory tract diseases have obtained timely control, and viral respiratory disease increases relatively, the clinical manifestation of viral respiratory disease can be varied, if it is slight to show as the most of symptoms of common cold and upper respiratory tract infection, bronchiolitis and pneumonia are often serious, can show as cough, spitting of blood persistence dyspnea, cyanosis anoxia etc., there is patchy shadow in the visible pulmonary of rabat, severe one is died because being choked to death, and what have is that respiratory and circulatory failure is dead.Therefore, CVB virus is very important in respiratory tract infection.But there is no effective antagonism medicine in the treatment for the CVB viral pneumonia at present.Can find Coxsackie B virus ribonucleic acid (CVB-RNA) in patient's blood WBC and make a definite diagnosis with high-tech PCR.Sophocarpine injection of the present invention has obvious therapeutical effect to the CVB viral pneumonia.Its mechanism of action is relevant with effective hematodinamics with the pharmacokinetics of sophocarpine uniqueness.The inventor uses sophocarpine in the test of Wistar mice pharmacokinetics, confirmation in lung (Lung) drug level considerably beyond blood concentration, and it is longer to hold time, and when blood drug level was 7.3350ug/ml, the concentration in the lungs tissue was 13.104ug/ml; When blood drug level was 4.787ug/ml, the concentration in lungs was 10.402ug/ml; When blood drug level was 2.8479ug/ml, the concentration in lungs was 5.124ug/ml; When blood drug level was 1.222ug/ml, the concentration in lungs was 2.509ug/m.And concentration can measure in 9～15 hours.Just due to such pharmacokinetics basis, favourable to the COXB virus of removing pulmonary, promote the recovery of pulmonary disease.
Because viral pneumonia due to COXB virus is coughed, panted, the respiratory symptom such as out of breath is obvious, respiratory secretions increases, sophocarpine has antitussive, antiasthmatic effect in Chinese medicine, in one embodiment of the invention, relevant hemodynamics to dog is studied, and finds that sophocarpine can make the lung capillary wedge that has raise press (PCWP) to drop to normally.In an example of the present invention, the 18mmHg of its PCWP before by administration drops to 12mmHg, and tends towards stability.The effect of sophocarpine also helps the resistance (SVR) that reduces the body circulation, by the 4732dyns/cm before medication
5Drop to 1579dyns/cm
5Body circulation resistance index (SVRI) is by the 2841dyns/cm before medication
5Cm
2Drop to 1579dyns/cm
5Cm
2And be conducive to pulmonary vascular resistance decline, by the 229dyns/cm before medication
5Decline is as for 57dyns/cm
5Pulmonary circulation index (PCRI) is by 115dyns/cm
5Cm
2Drop to 82 dyns/cm
5Cm
2
Thus, can improve pulmonary circulation, reduce pulmonary congestion, reduce preload, the afterload of heart, be conducive to the protection of heart and lungs function.The animal model of the Balb/c mice that the inventor infects by CBV, if normal group, virus control group and sophocarpine treatment group, take off neck after 10 days lethal, take out lungs (Lung) and do the pathology cut sections for microscopic examination, confirm that sophocarpine treatment group pulmonary lesion obviously alleviates, approach normally, illustrate that sophocarpine pneumonia due to treatment CVB is effective, pathological section such as Figure 28, Figure 29 and shown in Figure 30.
Claims (4)
1. sophocarpine or its officinal salt application in the medicine of the viral dilated cardiomyopathy that preparation treatment Coxsackie B virus causes, wherein, the structural formula of described sophocarpine is (I):
2. sophocarpine or its officinal salt application in the medicine of the viral dilated cardiomyopathy that preparation treatment Coxsackie B virus causes as described in claim 1, it is characterized in that: the officinal salt of described sophocarpine is bromate or the hydrochlorate of sophocarpine or the fluorate of sophocarpine of sophocarpine.
3. sophocarpine as claimed in claim 1 or its officinal salt application in the medicine of the viral dilated cardiomyopathy that preparation treatment Coxsackie B virus causes, it is characterized in that: the dosage form of described medicine is any dosage form of medically approving.
4. sophocarpine as claimed in claim 3 or its officinal salt application in the medicine of the viral dilated cardiomyopathy that preparation treatment Coxsackie B virus causes, it is characterized in that: the dosage form of described medicine is powder or injection or capsule or tablet or oral liquid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110257860 CN102349898B (en) | 2007-10-26 | 2007-10-26 | Application of sophocarpine to preparation of medicament for treating cardiovascular diseases caused by coxsackie virus B |
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| CN 200710047507 Division CN101416964B (en) | 2007-10-26 | 2007-10-26 | Use of sophocarpine in preparing medicine for treating diseases induced by coxsackie B virus |
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| CN108210495A (en) * | 2018-03-29 | 2018-06-29 | 上海壹志医药科技有限公司 | The new pharmaceutical uses of sophocarpine |
| CN116769733B (en) * | 2023-08-02 | 2023-11-03 | 中国医学科学院医学生物学研究所 | Coxsackie virus B group 4 strain and application thereof |
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| CN1303673A (en) * | 1999-10-26 | 2001-07-18 | 上海第二医科大学附属仁济医院 | Application of sophocarpine in preparation of medicine for curing coxsackievirus B myocarditis and its preparation method |
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