CN103565803A - Application of rupintrivir - Google Patents
Application of rupintrivir Download PDFInfo
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- CN103565803A CN103565803A CN201210269570.1A CN201210269570A CN103565803A CN 103565803 A CN103565803 A CN 103565803A CN 201210269570 A CN201210269570 A CN 201210269570A CN 103565803 A CN103565803 A CN 103565803A
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- coxsackie virus
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Abstract
The invention discloses application of rupintrivir, namely application of rupintrivir in preparation of a medicine used for treating diseased caused by enterovirus-71 type.
Description
Technical field
The present invention relates to field of medicaments, relate in particular to the purposes of a kind of Lu Ping Qu Wei.
Background technology
Enterovirns type 71 (EV71) belongs to Picornaviridae, and it is to cause the popular main pathogen of hand-foot-mouth disease (HFMD) in recent years.The infection of most of EV71 is self limiting, only can cause slight symptom as measles children.Yet in great outburst closely several times, some serious cases with neurological conditions frequently occur, these symptoms can worsen into cardiorespiratory failure fast.
Since California, USA reported first EV71 in 1969 infects, EV71 has occurred breaking out among a small circle and being identified in the U.S., Europe, Australia, Japan, Brazil and Malaysia.Within 1998, at TaiWan, China, there is an EV71 great outburst, caused 129,106 routine hand-foot-mouth disease and the serious case of 405 examples, therefore aroused the vigilant consciousness of the public to hand-foot-mouth disease.It should be noted that the popular death that has caused surpassing 20 people of an EV71 that occurs in Chinese Anhui Province between in March, 2008 to May.
Consider that EV71 infects relevant serious clinical manifestation, thereby prevent that in the urgent need to developing effective antiviral method EV71 from infecting morbidity and the death causing.At present, not can be used for clinically the direct antiviral drugs that serious EV71 infects.The symptoms such as heating, encephalitis and aseptic meningitis are alleviated by immunoglobulin in supportive medicine and vein.Although broad-spectrum antiviral medicament ribavirin is in the news, can reduce the mortality rate after 12-14 in age days ICR mouse infection EV71 of Mus, the dosage using (100mg/kg) approaches the adult mice LD50 value (220mg/kg/d) be in the news and far away higher than the dosage range (infection hepatitis C virus using dosage 10-16mg/kg/d grows up) of clinical recommendation.EV71 due to 95% infects and betides 4 years old following child, therefore uses the ribavirin of high dose may bring serious safety problem.Some other promising compound under study for action comprises that viral capsid binding molecule is as BPROZ series, albumen 2C inhibitor (metrifudil etc.), albumen 3A inhibitor (enviroxime etc.), 3D inhibitor (DTrip-22 and Aurintricarbosylic acid) and some other natural product (Raoul acid and ursolic acid etc.).Yet, thereby these compounds all do not carry out complete detection in animal model and clinical trial, do not set up its usefulness and safety spectrum.
Lu Ping Qu Wei (AG7088) is at first by the exploitation of Agouron Pharmaceuticals company, and it is a kind of irreversible HRV3C inhibitor.Yet whether Lu Ping Qu Wei has effect unknown to EV71 in animal body.In other words, not yet find so far the effective material of disease in vivo EV71 being caused.
Therefore, this area, in animal body can be effective in the urgent need to a kind of disease causing for EV71 is provided, and can really treat the material of this class disease.
Summary of the invention
The present invention aims to provide the new purposes of a kind of Lu Ping Qu Wei.
The invention provides the purposes of a kind of Lu Ping Qu Wei in preparing the medicine for the treatment of the disease being caused by enterovirus-71 type (enterovirus 71) and/or Coxsackie virus.
In another preference, described Coxsackie virus is Coxsackie virus-10 type or Coxsackie virus-16 type; More preferably, described Coxsackie virus is Coxsackie virus-16 type.
In another preference, described disease is hand-foot-mouth disease.
In another preference, described pharmaceutical preparation is solid preparation or liquid preparation; More preferably, described solid formulation comprises tablet, pill, capsule, powder, granule; Described liquid preparation comprises opacifiers, solution, suspending agent, syrup, drop.
In another preference, described preparation comprises oral formulations, intravenous formulations, intramuscular injectable formulations.
In another preference, described medicine contains has inhibiting active component and pharmaceutically acceptable carrier to enterovirus-71 type and/or Coxsackie virus; Preferably, described Coxsackie virus is Coxsackie virus-10 type or Coxsackie virus-16 type; More preferably, described Coxsackie virus is Coxsackie virus-16 type.
Accordingly, the invention provides a kind of disease causing for EV71 and/or Coxsackie virus, in animal body can be effective, can really treat the material of this class disease.
Accompanying drawing explanation
Fig. 1 has shown the bent Wei external activity of Lu Ping; Wherein,
(A)-(B) be that Lu Ping Qu Wei is to EV71(A) and CA16, CA10(B) dose-effect curve of the cytopathic effect that causes, (C) being the immunofluorescence of EV71 antigen, is (D) ability that RTCA analyzes the cytopathic effect of the bent Wei of Lu Ping or ribavirin antagonism EV71 induction.Fig. 2 has shown the antiviral activity of Lu Ping Qu Wei in body; Wherein.
(A) be Kaplan-Meier curve, (B) be clinical score, the obvious quadriplegia symptom (black arrow place) that C exists for the infecting mouse of taking for the 6th day after infecting, D detects β-actin content in EV71 RNA and cell within the 6th day, getting the capable real-time quantitative RT-PCR of intestinal, lung, limb muscle, brain stem and cardiac muscle (N=4-6) tissue of different group mices after infecting, and virus load is expressed as: – (Ct
eV71-Ct
actin)+10.
Fig. 3 has shown histology and the immunohistochemical analysis of each tissue, wherein,
Infect and after six days, get skeletal muscle (A-F), the parallel H & E dyeing of lung (G-L) and intestinal (M-O) specimen (A-C, G-I) and immunohistochemical analysis (D-F, J-O).G-I figure be every group of (N=6) representative picture (200 * amplify, other are 400 *).
The specific embodiment
Inventor, by animal model, has found that Lu Ping Qu Wei is for the therapeutic efficiency that has infected the animal of EV71.On this basis, completed the present invention.
Definition
As used herein, " Lu Ping Qu Wei (Rupintrivir) ", claims again AG-7088, refers to the compound of structure as shown in formula I, and its CAS registration number is 223537-30-2, and molecular formula is C
31h
39fN
4o
7:
As used herein, term " contain " or " comprising " comprised " comprising ", " substantially by ... form " and " by ... form ".
As used herein, term " effective dose " refers to and can produce function or amount active and that can be accepted by people and/or animal to people and/or animal.
As used herein, term " pharmaceutically acceptable carrier " refers to be used for the treatment of the carrier of agent administration, comprises various excipient and diluent.This term refers to some medicament carriers like this: they itself are not necessary active component, and after using, there is no undue toxicity.Suitable carrier is well known to those of ordinary skill in the art.In Remington ' s Pharmaceutical Sciences (Mack Pub.Co., N.J.1991), can find discussing fully about pharmaceutically acceptable excipient.On combination of Chinese medicine is learned, acceptable carrier can contain liquid, as water, saline, glycerol and ethanol.In addition, in these carriers, also may there is complementary material, as filler, disintegrating agent, lubricant, fluidizer, effervescent, wetting agent or emulsifying agent, correctives, pH buffer substance etc.
As used herein, term " medicine " or " pharmaceutical composition " can Alternates, all refer to by being applied to animal body, alleviate, improve or eliminate the material of animal body disease condition, wherein contain and have direct acting active component and pharmaceutically acceptable carrier to alleviating, improve or eliminating animal body disease.
Purposes
The present invention is used for the treatment of enterovirus-71 type by the pharmaceutical composition that contains Lu Ping Qu Wei and infects the disease causing.Active component in described compositions contains Lu Ping Qu Wei, can also contain the auxiliary activity composition that other has antivirus action, also can be by the Lu Ping Qu Wei for the treatment of effective dose with the Drug combination with antivirus action, to reduce the consumption of these medicines and to disappear the drug resistance to these medicines to virus.
In described pharmaceutical composition, can also contain pharmaceutically or acceptable carrier excipient or diluent in bromatology.This class carrier comprises (but being not limited to): saline, buffer, water, glycerol, ethanol and combination thereof.
Peroral administration solid composite medicament of the present invention can adopt the forms such as tablet, pill, capsule, powder, granule, drop.Active substance and at least one inert diluent of in these solid composites, having mixed one or more, for example, lactose, mannitol sugar, glucose, hydroxypropyl cellulose, microcrystalline Cellulose, starch, polyethylene pyrroles Anhui ketone, agar, pectin, aluminosilicate magnesium, magnesium aluminate.Also can make to contain the additive except inert agents in compositions according to common method, for example, the cosolvents such as lactose stabilized agent, glutamic acid or aspartic acid.Tablet or pill in this way, also can be as required, sucrose, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose or gastric solubility, enteric film in its outer wrap.
Peroral administration composition of liquid medicine comprises the opacifiers that allows on medicament, solution, suspending agent, syrup etc., and normally used inert diluent comprises Purified Water, ethanol.In said composition except inert diluent, also comprise wetting agent, suspending agent, etc. auxiliary agent, sweeting agent, correctives, aromatic and antiseptic.
Para-oral injection comprises sterile aqueous or non-aqueous solution agent, suspending agent and opacifiers.In aqueous solution agent and suspending agent, comprise distilled water for injection and normal saline.In non-aqueous solution agent and suspending agent, comprise propylene glycol, Polyethylene Glycol, cocoa butter, olive oil, Oleum Ricini, etc. vegetable oil.In these compositionss, also can comprise isotonic agent, antiseptic, wetting agent, emulsifying agent, dispersant, stabilizing agent, cosolvent.
Using method
In the present invention, the dosage of Lu Ping Qu Wei is not had to special restriction, available any suitable dosage.The type of carrier and quantity can be very not identical yet, and this depends on homoiothermic animal or people's kind, body weight and extent of damage of disease to be treated etc.Generally, suitable content is the 0.01%-99% that Lu Ping Qu Wei accounts for pharmaceutical composition gross weight, preferably 0.1%-99%.During as Drug therapy, the effective dosage ranges of Lu Ping Qu Wei is generally 0.01-800 mg/kg/day or higher, is preferably 0.1-500 mg/kg/day, and better is 0.5-200 mg/kg/day.
Dosage device comprises the bent Wei compound of a kind of Lu Ping, or the bent Wei compound of this Lu Ping and the formed mixture of other antiviral therapy agent.Dosage device also can contain diluent, filler, carrier etc.Dosage device is solid or gel form, as pill, tablet, capsule etc., or liquid form, their applicable oral administrations, topical or the intestines and stomach administration or intravenous administration, subcutaneous administration, muscle administration.
The pharmaceutical composition preparing can carry out administration by conventional route, comprising: intramuscular, intraperitoneal, intravenous, subcutaneous, Intradermal, oral or topical.In addition, the bent Wei of Lu Ping can also be used with together with the therapeutic agent of other treatment viral infection.
The above-mentioned feature that the present invention mentions, or the feature that embodiment mentions can combination in any.All features that this case description discloses can with any composition forms use, each feature disclosing in description, can anyly provide the alternative characteristics of identical, impartial or similar object to replace.Therefore except there being special instruction, the feature disclosing is only the general example of equalization or similar features.
Major advantage of the present invention is:
1, prove first the activity in vivo of the anti-EV71 of the bent Wei of Lu Ping.
2, the use amount scope safety of the anti-EV71 of the bent Wei of Lu Ping.
3, the bent Wei of Lu Ping can efficiently intercept the appearance of drug-resistant virus.
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment are only not used in and limit the scope of the invention for the present invention is described.The experimental technique of unreceipted actual conditions in the following example, conventionally according to normal condition or the condition of advising according to manufacturer.Unless otherwise indicated, otherwise all percent, ratio, ratio or umber by weight.
Unit in percent weight in volume in the present invention is well-known to those skilled in the art, for example, refer to the weight of solute in the solution of 100 milliliters.
Unless otherwise defined, the same meaning that all specialties of using in literary composition and scientific words and one skilled in the art are familiar.In addition, any method similar or impartial to described content and material all can be applicable in the inventive method.The use that better implementation method described in literary composition and material only present a demonstration.
Embodiment 1
The virus of using in embodiment, medicine, cell and mice source:
Human rhabdomyosarcoma (RD) cell culture is in the MEM culture medium containing 10% hyclone (containing glutamine, penicillin, streptomycin (GIBCO BRL)).
Lu Ping Qu Wei is purchased from Santa Cruz(SC-208317).
Enterovirns type 71 strain SHAPHC695F/SH/CHN/10(695F) be from infant oral cavity separation of one-year-old 10 months of Shanghai City public health clinical center, to obtain for 2010.
Coxsackie virus 16(CA16, strain 860F) and 10(CA10, strain 798F) from patient's oral cavity, separation obtains equally.
Specific pathogen free ICR mice (Charles River laboratory) is raised in Shanghai City public health clinical center animal center.Shanghai City public health clinical center animal center relevant regulations is followed in the treatment of animal.
Each test method in embodiment:
The infection of EV71 in RD cell
RD cell in 96 orifice plates infects respectively EV71 with 0.1MOI concentration, CA16 or CA10.Infection cell is containing variable concentrations DMSO(Dimethyl sulfoxide, dimethyl sulfoxide) or the culture medium of Lu Ping Qu Wei (Santa Cruz) in cultivate 3 days.Utilize CCK8 test kit (CCK-8, Dojindo, Japan) detect cell viability and draw dosage dependent reaction curve.
Referring to Fig. 1, wherein (A)-(B) Lu Ping Qu Wei is to EV71(A) and CA16, CA10(B) dose-effect curve of the cytopathic effect that causes.When Lu Ping Qu Wei (8-500nM) exists, with EV71 or CA16(MOI=0.1) infect RD cell and cultivate three days.CCK8 detect to obtain OD value, usings the logarithm of Lu Ping song Wei concentration as abscissa curve plotting.EC50 value detects according to method described in materials and methods.(C) immunofluorescence of EV71 antigen.RD cell infection EV71(MOI=0.01) and with DMSO or Lu Ping Qu Wei (30nM) process 4 days.Row immunofluorescence is also used Zeiss Axiovert fluorescence microscope.(D) RTCA analyzes the ability of the cytopathic effect of the bent Wei of Lu Ping or ribavirin antagonism EV71 induction.Be laid on the RD cell infection 100PFU EV71 virus of E-flat board and process with Lu Ping Qu Wei (30nM) or ribavirin (100 μ M), RTCA detects 8 days.Utilize cell index (CI) and time (hours) to draw.
Immunofluorescence
RD cell infection EV71(100pfu in 96 orifice plates) and with DMSO or the bent Wei of Lu Ping process (30nM).Infect latter 4 days, PBS washes cell, adds 3.7% paraformaldehyde to fix, and 0.1% TritonX-100 penetrates.Utilize after the PBS sealing containing 5% hyclone, the antiserum (1:200 dilution) that utilizes EV71 to infect cured person carries out primary antibodie hatches, and room temperature 2 hours is hatched two anti-(goat anti-human igg-FITC, Sigma 1:500 dilutions) subsequently.After washing cell, with Zeiss Axiovert inverted microscope, observe.In order to ensure the specificity of result, the RD cell not infecting dyes as negative control simultaneously.
Real-time cell analysis (RTCA)
Adopt real-time cell analysis (RTCA) system.This system can be passed through the microelectronic biosensor detecting electrode impedance of Tissue Culture Plate bottom with the growth of monitoring cell.RTCA system can directly be monitored cell growth process and do not needed to participate in labelling.RD cell is by every hole 1.0 * 10
4the 695F virus that is laid in 96 hole E-flat boards and infects 1000pfu.After 24 hours, change culture medium and Lu Ping Qu Wei (30nM) and ribavirin (100 μ M) are added in respective aperture.Cell culture is also observed 8 days.Upgrowth situation is described with growth index and is detected the state of the common showed cell of data with the electronic cell sensor of E-flat board bottom.
In mice, EV71 infects
With 10
6the EV71(695F strain of PFU) inject the Mus young Mus (2.0-2.3g) in 2 days ages, lumbar injection 5-15 μ l80 μ M or 800 μ M are dissolved in the Lu Ping Qu Wei of DMSO subsequently.Press the injection of 0.1mg/kg dosage for 13, press the injection of 1mg/kg dosage for another 13, the DMSO of matched group (N=14) injection same volume.Continuous 10 day every day injectable drug.Observe the morbidity of infecting mouse and dead sign every day.Ill degree to mice mark (0, health; 1, slow in action; 2, hind leg is weak; 3, single acroparalysis; 4, two acroparalysis; 5, death) in order better to show the recovery situation of survival mice, dead mice is only credited to average mark once when finding for the first time.
Referring to Fig. 2, the ICR mouse peritoneal in 2 days ages of Mus inoculation EV71 and with DMSO(N=13) and Lu Ping Qu Wei (0.1mg/kg, N=13,1mg/kg, N=14) processing, result is shown as Kaplan-Meier curve (A) and clinical score (B).Dead mice is only credited to average mark once when finding for the first time.C figure takes for after infecting the 6th day, and Image Display DMSO group infecting mouse exists obvious quadriplegia symptom (black arrow place).D figure gets different groups (A, DMSO group on the 6th day after infecting; B, the bent Wei 0.1mg/kg of Lu Ping group; C, the bent Wei 1mg/kg of Lu Ping group) the capable real-time quantitative RT-PCR of intestinal, lung, limb muscle, brain stem and cardiac muscle (N=4-6) tissue of mice detects β-actin content in EV71 RNA and cell.Virus load is expressed as: – (Ct
eV71-Ct
actin)+10.
Histology and immunohistochemical analysis
Various mouse tissues carry out conventional hematoxylin-eosin staining.3.7% formaldehyde is fixed, paraffin embedding 4um section.For the SABC of EV71 antigen, skeletal muscle, lung and intestinal tissue are embedded in OCT compound freezing in liquid nitrogen.Utilizing Leica CM1800 cryostat to obtain cervical region thin layer cuts into slices on the coated slide of poly-D-lysine and further with 3.7% paraformaldehyde, fixes.With the PBS containing 1.5% horse serum, seal, carry out primary antibodie subsequently hatch (Millipore, MAB979,1:500 dilution) by the monoclonal antibody of anti-EV71 VP1,4 ℃ are spent the night.These slides continue to hatch with biotinylated anti-mouse antibody, supplement Vector M.O.M reagent and ABC reagent in Vectastain ABC-peroxidase test kit (Vector laboratories) in antibody.Slide adds DAB substrate and uses Mayer ' s haematoxylin redyeing.In parallel laboratory test, be provided with strict contrast (not infecting each respective organization section of EV71) to guarantee the specificity of result.
The real-time quantitative reverse transcription PCR of viral RNA
Come from the muscle of different disposal group mice, intestinal, heart, lung and brain stem obtain for the 6th day after infection.Utilize the bead (Biospec Products) of 1mm, containing in the culture medium of 2% hyclone, tissue is being ground to form to microgranule of uniform size.Utilize QIAamp QIAXtractor Virus reagent(Qiagen, 950207) extracting RNA.Use the method amplification VP1 fragment of one-step method reverse transcription PCR test kit (one step Prime Script RT-PCR kit, Takara, Dalian, DRR064A) and real-time quantitative PCR.As internal reference, contrast is also amplified the beta-actin of mice simultaneously.Virus load is represented as :-(Ct
eV71-Ct
actin)+10
Data statistics
Data represent with average ± standard deviation.Utilize Mantel-Cox test to compare survival rate.
In Graphpad Prims software, utilize nonlinear regression (variable slope) analysis to enter EC50 value
Row is estimated.One-Way ANOVA test is for comparing two or more sets viral RNA content differences.
Mann-Whitney U test is for the difference of viral RNA content between comparing two groups.
The result of each test in embodiment
In cell line, the bent Wei of Lu Ping can effectively be removed copying of EV71
First in cell culture model, assessed the antiviral activity of Lu Ping Qu Wei.Can cause the 0.1MOI EV71 virus 695F strain infection cell of a large amount of cytopathic effecies, after inoculation, add Lu Ping Qu Wei (8nM to 0.5uM) and observe CPE in infection in latter 72 hours.Lu Ping Qu Wei can resist the cell death (EC50=14nM 95%CI 12.16-16.33nM, Figure 1A) of EV71 induction completely in relatively low concentration.CA16 and CA10 are two kinds of viruses that can cause HFMD that circulate extensively, and have also assessed the activity of the bent Wei opposing CA16 of Lu Ping and CA10.The bent Wei of Lu Ping can suppress the propagation of CA16 and CA10 equally, and EC50 is respectively 15nM and 17nM(Figure 1B).Detected in addition when Lu Ping Qu Wei (30nM) exists the expression of EV71 VP1 albumen in the RD cell infecting with 0.01MOI EV71.Immunofluorescence result is presented at EV71 and inoculates latter 48 hours viral massive duplications, yet does not substantially observe obvious signal after adding Lu Ping Qu Wei (30nM).The usefulness of Lu Ping Qu Wei that we have also further utilized RTCA system evaluation.As shown in Fig. 1 D, the upgrowth situation of RD cell (blueness) of processing through the bent Wei of 30nM Lu Ping is suitable with the upgrowth situation of non-infected cells (green), yet the ribavirin of 100 μ M (pink) is processed the cytopathic effect (redness) that can only partly save EV71 induction.
In children's mouse model, the bent Wei of Lu Ping can effectively be saved paralysis and the death that EV71 causes
Then in young mouse model, detected the antiviral activity of Lu Ping Qu Wei.After lumbar injection EV71 virus 695F strain (10E6PFU/ Mus), inject Lu Ping Qu Wei (0.1mg/kg or 1mg/kg) continuous 10 day every day, and matched group is injected DMSO with same volume.In this model, if treated with medicaments not can be observed most of young Mus in the 4th day after infection and occur the symptom that hind leg is weak.Several days subsequently, the situation of infecting mouse further worsened as hind leg or front acroparalysis (single limb or two limb, Fig. 2 C, upper figure, arrow place) and within 6-10 days after infection, reaches peak (Fig. 2 B).Yet the bent Wei Jun of Lu Ping of two kinds of dosage can significantly alleviate these symptoms (Fig. 2 C figure below), its clinical marking is significantly lower than DMSO group (Fig. 2 B, p<0.01 Wilcoxon rank test).Mortality rate for infecting mouse; 0.1mg/kg dosage injection Lu Ping Qu Wei is enough to almost completely (90.9% survival; Fig. 2 A) protection mice avoids the death that viral infection causes, yet DMSO group finally only has 38.5% survival (p=0.0063 Mantel-Cox test) what observe.The bent Wei of 1mg/kg dosage injection Lu Ping further do not improve overall survival (83.3% with 38.5%DMSO group, p=0.012).Further analyze the content that infects viral RNA in rear the 6th day different tissues.The 6th day skeletal muscle inner virus carrying capacity the highest (DMSO group skeletal muscle inner virus RNA is 2000-4000 times in its hetero-organization) in selected tissue after infecting.Consistent with the symptom of observing, after the bent Wei administration of Lu Ping, viral RNA content is all decreased significantly in each tissue: intestinal, p<0.0001; Lung, p<0.01; Muscle, p<0.05; Brain stem, p<0.01; Cardiac muscle, p<0.0001(One-Way ANOVA).In addition, the Lu Ping Qu Wei of 1mg/kg dosage has strengthened effect (Fig. 2 D, intestinal, lung and cardiac muscle, the p<0.01 of its inhibition EV71 RNA than the bent Wei Xianzhu of the Lu Ping of 0.1mg/kg dosage; Brain stem, p<0.05, Mann-Whitney U test).Though the minimizing of viral RNA clearly but there is no in its hetero-organization significantly, especially there is no significant difference between DMSO group and 0.1mg/kg dosed administration group in muscle.Yet immunofluorescence result shows that the expression of virus VP 1 albumen is significantly inhibited.
Histological observation
Detected histology's performance of each tissue of infecting mouse.When infecting mouse is fallen ill, heart and spinal cord significantly do not damage and/or inflammation (data do not show).By contrast, there is the necrotizing myositis (Fig. 3 B) with inflammatory infiltration in limb muscle.Although some slight damages still can be observed, the bent Wei of 0.1mg/kg Lu Ping is processed the integrity (Fig. 3 C) that has significantly strengthened limb muscle structure.The immunohistochemical staining of VP1 has confirmed the result of RT-PCR, at DMSO group virus protein the most active (Fig. 3 E400 *, black arrow place) in the bright skeletal muscle of signal list strongly and widely, in matched group without this phenomenon (Fig. 3 D400 *).In muscle, the expression of VP1 is subject to the remarkable inhibition (Fig. 3 F 200 * and Fig. 3 I400 *) of Lu Ping Qu Wei (0.1mg/kg).DMSO processes pulmonary's moderate inflammation infiltration of mice and the symptom of alveolar septum multiviscosisty is eased after processing with Lu Ping Qu Wei (0.1mg/kg).In addition, the dyeing of EV71 VP1 is significantly but more faint in alveolar (Fig. 3 K) and casing slime and basement membrane (Fig. 3 N, black arrow place).To sum up result shows that the bent Wei processing of Lu Ping significantly reduces the expression (Fig. 3 L, O) of virus antigen in each tissue.
While adopting low dosage (0.1mg/kg), the bent Wei of Lu Ping can almost completely be resisted EV71 and infect the dead mouse causing, this result has proved the interior resisting virus effect of Lu Ping Qu Wei completely.Because the child that EV71 infects is more desirable to more responsive bent this characteristic of Wei of Lu Ping that makes of drug toxicity than adult.It is the most active at skeletal muscle that SABC and Real-time PCR Analysis show that EV71 infects, and serious necrotizing myositis can occur for it.The bent Wei medication of Lu Ping can suppress consumingly virus replication and improve muscular tissue and limb activity.Even in 0.1mg/kg group, virus antigen is expressed and is also subject to strong inhibition.
Directly the antiviral drugs of targeting viral enzyme usually produces drug resistance.In fact, Lu Ping Qu Wei external selection of drug resistance in HRV2 and 14 is in the news.Monamino acid sudden change only can cause the moderate decline of sensitivity and significant drug-resistant phenotype often needs the sudden changes of three above key amino acid residues.Inventor attempts estimating the medicament-resistant mutation of EV71, yet 12 continuous gradient screenings that the bent Wei concentration of Lu Ping rises gradually all do not produce activated mutated viruses strain.This result shows that the bent Wei of Lu Ping can efficiently intercept the appearance of drug-resistant virus, and this embodies again its another clinical advantage.
Preparation is containing the bent Wei freeze dried powder of Lu Ping
| Formula | Content (g) |
| Lu Ping Qu Wei | 25 |
| Mannitol | 125 |
| Water for injection | 2500 |
| Make the bent Wei freeze dried powder of Lu Ping | 0.025/ bottle of X1000 bottle |
According to recipe quantity precision, take raw material, recipe quantity mannitol is dropped into, add the water for injection of the total amount 80% of approximately writing out a prescription to be stirred to dissolve complete, obtain settled solution, the needle-use activated carbon that adds 0.1% (g/ml), stirs, standing approximately 10 minutes, through 0.45 μ m filtering with microporous membrane, benefit adds to the full amount of water for injection.Again through 0.22 μ m filtering with microporous membrane, measure pH value and content, qualified after, quantitative filling, lyophilization, fills nitrogen, tamponade, jewelling lid, labeling, packing, it is qualified to inspect by random samples, gets product.
The foregoing is only preferred embodiment of the present invention, not in order to limit essence technology contents scope of the present invention, essence technology contents of the present invention is to be broadly defined in the claim scope of application, any technology entity or method that other people complete, if defined identical with the claim scope of application, also or a kind of change of equivalence, all will be regarded as being covered by among this claim scope.
Claims (10)
1. the purposes in the medicine of the disease that a Lu Ping Qu Wei is caused by enterovirus-71 type (enterovirus 71) and/or Coxsackie virus in preparation treatment.
2. purposes as claimed in claim 1, is characterized in that, described Coxsackie virus is Coxsackie virus-10 type or Coxsackie virus-16 type.
3. purposes as claimed in claim 2, is characterized in that, described Coxsackie virus is Coxsackie virus-16 type.
4. purposes as claimed in claim 1, is characterized in that, described disease comprises hand-foot-mouth disease.
5. purposes as claimed in claim 1, is characterized in that, described pharmaceutical preparation is solid preparation or liquid preparation.
6. purposes as claimed in claim 5, is characterized in that, described solid formulation comprises tablet, pill, capsule, powder, granule; Described liquid preparation comprises opacifiers, solution, suspending agent, syrup, drop.
7. purposes as claimed in claim 5, is characterized in that, described preparation comprises oral formulations, intravenous formulations, intramuscular injectable formulations.
8. purposes as claimed in claim 1, is characterized in that, described medicine contains has inhibiting active component and pharmaceutically acceptable carrier to enterovirus-71 type and/or Coxsackie virus.
9. purposes as claimed in claim 8, is characterized in that, described Coxsackie virus is Coxsackie virus-10 type or Coxsackie virus-16 type.
10. purposes as claimed in claim 9, is characterized in that, described Coxsackie virus is Coxsackie virus-16 type.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210269570.1A CN103565803A (en) | 2012-07-31 | 2012-07-31 | Application of rupintrivir |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210269570.1A CN103565803A (en) | 2012-07-31 | 2012-07-31 | Application of rupintrivir |
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| CN105688216A (en) * | 2016-01-29 | 2016-06-22 | 中国科学院上海巴斯德研究所 | Composition for treating enterovirus infection and drug combination method |
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| WO2011160191A1 (en) * | 2010-06-25 | 2011-12-29 | Biota Scientific Management Pty Ltd | Compound for the treatment of enteroviruses |
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| WO2011160191A1 (en) * | 2010-06-25 | 2011-12-29 | Biota Scientific Management Pty Ltd | Compound for the treatment of enteroviruses |
Non-Patent Citations (2)
| Title |
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| 何静等: "抗肠道病毒71型药物研究进展", 《医学综述》, vol. 17, no. 9, 31 May 2011 (2011-05-31), pages 1281 - 1283 * |
| 赵晓光等: "肠道病毒检测及其抗病毒药物研究进展", 《中国公共卫生》, vol. 23, no. 3, 31 March 2007 (2007-03-31), pages 375 - 377 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105688216A (en) * | 2016-01-29 | 2016-06-22 | 中国科学院上海巴斯德研究所 | Composition for treating enterovirus infection and drug combination method |
| CN105688216B (en) * | 2016-01-29 | 2019-01-25 | 中国科学院上海巴斯德研究所 | Composition and combination drug for treating enterovirus infection |
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