CN104510747A

CN104510747A - New medicinal application of iridoid glycoside

Info

Publication number: CN104510747A
Application number: CN201310462725.8A
Authority: CN
Inventors: 樊向德; 陈鸣珍
Original assignee: Individual
Current assignee: Individual
Priority date: 2013-09-30
Filing date: 2013-09-30
Publication date: 2015-04-15
Anticipated expiration: 2033-09-30
Also published as: CN104510747B

Abstract

The invention belongs to the field of medicines and particularly relates to a new application of an iridoid glycoside compound, genipin-1-[beta]-D-gentiobioside, and a composition composed by compounding the genipin-1-[beta]-Dgentiobioside with other components as effective components for preparing antiviral drugs, antibacterial drugs, fever alleviating drugs, inflammation resistant drugs and anti-oxidizing drugs. The iridoid glycoside compound and the composition can be used for treating acute respiratory infection, flu, pneumonia, viral hepatitis type B, herpes zoster, herpes viral keratitis, viral myocarditis, Epstein-Barr virus infection, retrovirus infectious diseases and bacterial infectious diseases.

Description

A kind of new medicine use of iridoid glycoside

Technical field

The invention belongs to field of medicaments, be specifically related to a kind of iridoid glycoside compound--genipin-1-β-D gentiobioside with cape jasmine and the compositions that formed with other components compatibilities thereof for the preparation of antiviral, antibacterial, bring down a fever, novelty teabag in antiinflammatory, anti-oxidation medicine.

Background technology

Acute respiratory infection is not only common clinical, frequently-occurring disease, especially the appearance of SARS, crowd's bird flu, H1N1 influenza and H7N9 influenza in recent years makes the mankind be among pandemic danger at any time, and existing anti-infectives and vaccine can not successfully manage the source of infection of constantly change.Therefore, the medicine finding effective and safe is the significant problem of 21 century facing mankind.

Hepatitis B is worldwide popular, is a worldwide public health difficult problem, has that infectiousness is strong, route of transmission is complicated, popular wide, sickness rate high.China is that HBV height infects, the high popular country of hepatitis B.WHO divides China into chronic viral hepatitis B hotspot." People's Republic of China's health statistics yearbook in 2012 " discloses: in 28 kinds of infection disease notification morbidities and death toll statistics, viral hepatitis stands above the 5th, and China is used for the treatment of hepatitis B and diseases induced expense thereof every year up to 300-500 hundred million yuans.

The medicine of existing treatment hepatitis B mainly contains: plain interferon α-2b, lamivudine, Peg-IFN alpha-2b α-2a, adefovirdipivoxil, Entecavir (Entecavir) tenofovir disoproxil (TDF) etc.The advantage of interferon is that the course for the treatment of is more fixing, and higher to transaminase, virus levels is not bery high patient's comparitive study is good, and E antigen conversion ratio is also higher.Its shortcoming is also apparent, need injection, and spray interferon and can produce the untoward reaction such as leukopenia, heating, bone marrow depression, parainfluenza sample symptom, although there is the fixing course for the treatment of, subset of patients can change into " small three positive " from " great three positive ", but major part still can not be turned out cloudy.Nucleotide drug is oral drugs, taking convenience, there is no direct toxic and side effects, and comparitive study is quick, and after taking medicine, virus can reduce soon.Its shortcoming is that the course for the treatment of is long, and the course for the treatment of is not determine especially, and somebody needs 2 years, and what have may be longer, needs Long-term taking medicine to treat.A shortcoming is also had to be the problem that can produce drug resistance in the process of Long-term taking medicine.Therefore, the clinical at present therapeutic agent for hepatitis B be still badly in need of safely and effectively, there is Anti-HBV activity effect.

The infection of Epstein-Barr virus in crowd has quick ascendant trend.Child infects rear most non-evident sympton, or causes mild pharyngitis and upper respiratory tract infection.Adolescence generation primary infection, about has 50% to occur infectious monocytosis.Propagate mainly through saliva, also can through post-transfusion.Epstein-Barr virus, in the internal breeding of pars oralis pharyngis epithelial cell, then infects bone-marrow-derived lymphocyte, and these cells enter blood circulation in a large number and cause systemic infection, and can hide for a long time in human lymphoid's tissue.When body's immunity is low, likely cause kinds of tumors.

About the treatment of ebv infection, acycloguanosine (AC) and gancyclovir (DHPG) can press down EBV and copy, and all have certain curative effect.Have two kinds of vaccines to come out at present, one of them is the vaccine vaccine of expressing EBVgp320 and HBsAg while China's gene engineering method builds, and emphasis is used in high risky area of nasopharyngeal carcinoma.Another is purified virus gp320 memebrane protein vaccine, just doing inoculation on a small scale in Britain, whether can reduce the sickness rate of infectious monocytosis to observing this vaccine.Therefore, the medicine for the treatment of ebv infection will have market prospect and clinical value widely.

Iridoid is the cyclic monoterpenes derivant containing cyclopentane structure unit, is distributed widely in nature, and the plants such as Rubiaceae, Oleaceae, Gentianaceae and Scrophulariaceae are many containing this constituents.Iridoid has multiple pharmacologically active, as blood pressure lowering, hepatic cholagogic, antitumor, antiinflammatory, neuroprotective, treatment diabetes and complication etc.

Genipin-1-β-D gentiobioside with cape jasmine is iridoid effective constituents contained in the plants such as Fructus Gardeniae, and this compound also can be obtained by synthetic.Notification number is the method that the Chinese patent of CN1706858 discloses from Fructus Gardeniae separation and purification genipin-1-β-D gentiobioside with cape jasmine; Chinese patent CN102000102A discloses the application of genipin-1-β-D gentiobioside with cape jasmine in preparation treatment heart failure disease medicine; " Pharmacology and Clinics of Chinese Materia Medica " the 2nd phase in 2013, the 39th page to 41 pages report genipin-1-β-D gentiobioside with cape jasmines were by improving the contractile function of heart and reducing its preload, improved the heart failure that pentobarbital sodium causes; " new Chinese medicine and clinical pharmacology " the 1st phase in 2009 the 8th page is reported to the 10th page, and the Fructus Gardeniae total iridoid glycosides containing genipin-1-β-D gentiobioside with cape jasmine can intervene the inflammatory reaction after cerebral hemorrhage, stops Neuron Apoptosis.

Summary of the invention

For this reason, technical problem to be solved by this invention be to provide a kind of genipin-1-β-D gentiobioside with cape jasmine (No. CAS: 29307-60-6) and combination thereof for the preparation of antiviral, antibacterial, bring down a fever, antiinflammatory, antioxidation, and be used for the treatment of the novelty teabag of acute respiratory infection, influenza, pneumonia, tracheitis, bronchitis, cough, hepatitis B, herpes zoster, simplex keratitis keratitis, viral myocarditis, ebv infection, retroviral infection disease and bacterial infection disease medicine.

Above-mentioned technical problem is solved by following scheme:

One have antiviral, antibacterial, bring down a fever, antiinflammatory, antioxidation medicine, described medicine with genipin-1-β-D gentiobioside with cape jasmine for active component.

The invention also discloses a kind of medicine being used for the treatment of acute respiratory infection, influenza, pneumonia, tracheitis, bronchitis, cough, hepatitis B, herpes zoster, simplex keratitis keratitis, viral myocarditis, ebv infection, retroviral infection disease and bacterial infection disease, described medicine with genipin-1-β-D gentiobioside with cape jasmine for active component.

The active component of described medicine consists of the mixture of genipin-1-β-D gentiobioside with cape jasmine or genipin-1-β-D gentiobioside with cape jasmine and geniposide (No. CAS: 24512-63-8) and/or shanzhiside methyl ester (No. CAS: 64421-28-9).

Preferably, the active component of described medicine consists of:

Genipin-1-β-D gentiobioside with cape jasmine 1-8 weight portion, geniposide 1-8 weight portion; Or

Genipin-1-β-D gentiobioside with cape jasmine 0.5-6 weight portion, geniposide 0.5-6 weight portion, shanzhiside methyl ester 0.2-4 weight portion.

Same, the active component of described medicine consists of:

Genipin-1-β-D gentiobioside with cape jasmine 1 weight portion, geniposide 1 weight portion; Or

Genipin-1-β-D gentiobioside with cape jasmine 3 weight portion, geniposide 1 weight portion; Or

Genipin-1-β-D gentiobioside with cape jasmine 6 weight portion, geniposide 1 weight portion; Or

Genipin-1-β-D gentiobioside with cape jasmine 1 weight portion, geniposide 3 weight portion; Or

Genipin-1-β-D gentiobioside with cape jasmine 1 weight portion, geniposide 6 weight portion; Or

Genipin-1-β-D gentiobioside with cape jasmine 1 weight portion, geniposide 1 weight portion and shanzhiside methyl ester 0.5 weight portion.

Described medicine makes clinical or pharmaceutically acceptable tablet, capsule, granule, aqueous injection, injectable powder, lyophilized injectable powder, spray, suppository.

The invention also discloses said medicine prepare antiviral, antibacterial, bring down a fever, application in antiinflammatory, anti-oxidation medicine.

Described antiviral refers to resisiting influenza virus, against parainfluenza virus, anti respiratory syncytial virus, rhinovirus, anti-adenovirus, anti-hepatitis virus, anti-Coxsackie virus, anti-Echovirus, anti-herpesvirus, anti EB virus and antiretroviral.

The invention also discloses the application of said medicine in preparation treatment acute respiratory infection, influenza, pneumonia, tracheitis, bronchitis, cough, hepatitis B, herpes zoster, simplex keratitis keratitis, viral myocarditis, ebv infection, retroviral infection disease and bacterial infection disease medicine.

The route of administration of described medicine comprises acceptable oral administration, drug administration by injection, intravenous drip administration, sublingual administration, spraying suction and rectally clinically.

The invention also discloses a kind of Fructus Gardeniae extract as active component for the preparation of antiviral, antibacterial, bring down a fever, the application of antiinflammatory, anti-oxidation medicine, described Fructus Gardeniae extract is prepared by the following method::

Get Fructus Gardeniae 1 weight portion, chopping, with the alcohol dipping 0.5-1 hour of 5--10 parts by volume 30-60%, reflux, extract, 1-2 hour, again with the 30-60% alcohol reflux 0.5-2 hour of 4-8 parts by volume, filter, merging filtrate,-9Pa, 40-80 DEG C of decompression recycling ethanol, obtain extracting solution, by extracting solution in NKA-2 macroporous resin loading, with 1-4 times of column volume water elution, eluent is in S-8 macroporous resin loading, with 1-4 times of column volume, concentration is the ethanol elution of 20-50%, eluent decompression recycling ethanol, continue concentrating under reduced pressure, concentrated solution dehydrated alcohol process, genipin-1-β-D gentiobioside with cape jasmine must be contained, geniposide two kinds of compositions are main extract, or

Get Fructus Gardeniae 1 weight portion, chopping, with the alcohol dipping 0.5-1 hour of 5--10 parts by volume 30-60%, reflux, extract, 1-2 hour, again with the 30-60% alcohol reflux 0.5-2 hour of 4-8 parts by volume, filter, merging filtrate,-9Pa, 40-80 DEG C of decompression recycling ethanol, obtain extracting solution, by extracting solution in ADS-17 macroporous resin loading, with 1-4 times of column volume water elution, eluent is in LSA308 macroporous resin loading, with 1-4 times of column volume, concentration is the ethanol elution of 20-50%, , eluent decompression recycling ethanol, continue concentrating under reduced pressure, concentrated solution dehydrated alcohol process, genipin-1-β-D gentiobioside with cape jasmine must be contained, geniposide and shanzhiside methyl ester three kinds of compositions are main extract, or

Get Fructus Gardeniae 1 weight portion, chopping, Soakage extraction 1-5 hour is stirred with the room temperature water of 7-11 parts by volume, 1-3 hour is extracted again with 5-10 parts by volume room temperature water, filter, merging filtrate, obtain extracting solution, by extracting solution in ADS-17 macroporous resin loading, with 1-4 times of column volume water elution, eluent is in NKA-2 macroporous resin loading, with 1-4 times of column volume, concentration is the ethanol elution of 20-50%, again in NKA-2 macroporous resin loading after eluent decompression recycling ethanol, continue with 1-4 times of column volume, concentration is the ethanol elution of 30-70%, eluent decompression recycling ethanol, continue concentrating under reduced pressure, concentrated solution dehydrated alcohol process, genipin-1-β-D gentiobioside with cape jasmine must be contained, geniposide and shanzhiside methyl ester three kinds of compositions are main extract, or

Get Fructus Gardeniae 1 weight portion, chopping, with the water room temperature immersion 0.5-1 hour of 7-11 parts by volume, decoct and extract 1-3 hour, use 5-10 parts by volume soak by water 0.5-2 hour again, filter, merging filtrate, obtain extracting solution, by extracting solution in LSA308 macroporous resin loading, with 1-3 times of column volume water elution, eluent is in S-8 macroporous resin loading, with 1-4 times of column volume, concentration is the ethanol elution of 20-50%, again in NKA-2 macroporous resin loading after eluent decompression recycling ethanol, continue with 1-4 times of column volume, concentration is the ethanol elution of 30-70%, eluent decompression recycling ethanol, continue concentrating under reduced pressure, concentrated solution acetone treatment, genipin-1-β-D gentiobioside with cape jasmine must be contained, geniposide and shanzhiside methyl ester three kinds of compositions are main extract,

The pass of described weight portion and parts by volume is the relation of g/ml.

The invention also discloses Fructus Gardeniae extract that said method prepares as active component for the preparation of the application in treatment acute respiratory infection, influenza, pneumonia, tracheitis, bronchitis, cough, hepatitis B, herpes zoster, simplex keratitis keratitis, viral myocarditis, ebv infection, retroviral infection disease and bacterial infection disease medicine.

Medicine of the present invention directly can be prepared by known compound genipin-1-β-D gentiobioside with cape jasmine, geniposide and/or shanzhiside methyl ester; Or utilize the method for disclosed separation and purification genipin-1-β-D gentiobioside with cape jasmine, geniposide and the shanzhiside methyl ester from Fructus Gardeniae of Chinese patent CN1706858A, and the preparation that is directly used.

Technique scheme of the present invention has the following advantages compared to existing technology:

1, the mixture of described genipin-1-β-D gentiobioside with cape jasmine and itself and geniposide and/or shanzhiside methyl ester, to H1N1 Influenza B virus strain infection ICR murine pneumonia model, the Lung Exponent of infection animal can be reduced, reduce mortality of animals, extend the time-to-live, reduce and infect virus load in rear lung tissue;

2, the mixture of described genipin-1-β-D gentiobioside with cape jasmine and itself and geniposide and/or shanzhiside methyl ester, to parainfluenza virus, respiratory syncytial virus infection BALB/C mice pulmonary inflammation model, can reduce the Lung Exponent of infection animal;

3, the mixture of described genipin-1-β-D gentiobioside with cape jasmine and itself and geniposide and/or shanzhiside methyl ester, to the scorching model of herpesvirus infection rabbit cornea, can alleviate keratopathy degree;

4, the mixture of described genipin-1-β-D gentiobioside with cape jasmine and itself and geniposide and/or shanzhiside methyl ester, ICR mouse core myositis virus model is infected to CoXB3 virus N acy strain, mortality of animals can be reduced, alleviate cardiac index, reduce AST, LDH content in cardiac muscular tissue, and alleviate the pathology damage degree of viral infection rear myocardium tissue;

5, the mixture of described genipin-1-β-D gentiobioside with cape jasmine and itself and geniposide and/or shanzhiside methyl ester, to DHB model, can reduce DHBV-DNA and DHBsAg titre in serum;

6, the mixture of described genipin-1-β-D gentiobioside with cape jasmine and itself and geniposide and/or shanzhiside methyl ester, to CNE-2Z cell Model in Nude Mice, can the growth of Tumor suppression;

7, in testing in vitro, the mixture of described genipin-1-β-D gentiobioside with cape jasmine and itself and geniposide and/or shanzhiside methyl ester, all has obvious inhibitory action to the strain of H1N1 Influenza B virus, PR8 strain, PIV-1, HSV-1, HSV-2, RSV, CoxB3, CoxB5, Ad3, Ad7, rhinovirus;

8, the mixture of described genipin-1-β-D gentiobioside with cape jasmine and itself and geniposide and/or shanzhiside methyl ester, to the staphylococcus aureus (type strain 26003 of In vitro culture, 331, 360, 361), Staphylococcus albus (26101), staphylococcus epidermidis (type strain, 104, 108), beta hemolytic streptococcus (10, 12), streptococcus pneumoniae (31001, 160, 162, 163) escherichia coli (type strain 44113, 107, 178), bacillus pyocyaneus (10102, 209, 210, 211), Candida albicans (type strain), pneumobacillus (11, 14), micrococcus catarrhalis (29108) growth in vitro all has obvious inhibitory action,

9, the mixture of described genipin-1-β-D gentiobioside with cape jasmine and itself and geniposide and/or shanzhiside methyl ester, has obvious antipyretic effect to rabbit fever models caused by lipopolysaccharide;

10, the mixture of described genipin-1-β-D gentiobioside with cape jasmine and itself and geniposide and/or shanzhiside methyl ester, significantly can reduce mouse lung tissue MDA, INF-r and TNF-a content after influenza infection, reduce glacial acetic acid and cause mouse peritoneal capillary permeability, show that there is obvious antiinflammatory action.

11, each administration treated animal gives (4 hours, interval) after each medicinal liquid 2 times of Cmax by 0.2ml/10g body weight lumbar injection, and death does not appear in each treated animal, and body weight increases normal, shows that 9 tested materials are without overt toxicity effect.

12, give rat with the dosage gavage of 15 times to 70 times of the clinical plan consumption of people, continuous 4 weeks for reagent 1,2,7,8,9, recover 2 weeks after drug withdrawal, rat oral gavage gives for reagent 1, No. 2 20g/kg/d, 10g/kg/d, 5g/kg/d; For reagent No. 7 15g/kg/d, 7.5g/kg/d, 3.25g/kg/d and for reagent 8, No. 9 25g/kg/d, 12.5g/kg/d, 6.25g/kg/d, continuous 1 month, continue observation after drug withdrawal 2 weeks.Result shows: during administration, the general activity of animal is normal, the mental status is good; Defecation is normal, occurs without vomiting; Hair smoothing, have no obscission; No abnormality seen bleeding tendency.Body weight, food-intake, electrocardio are had no significant effect; The ponderal index of every biochemical indicator, peripheral hemogram and important organ all fluctuates within normal range; Each internal organs macroscopy is showed no obvious pathological change; Pathological study also has no the pathomorphology change caused by medicine; Have no drug-induced toxicity.

13, be that test medicine treatment acute respiratory infection Clinical efficacy and safety are evaluated in contrast with placebo.Dosage regimen is each 2, every day 3 times, within 3 days, is 1 course for the treatment of.Shown by double-blind trial result, in cure rate, total obvious effective rate, total effective rate etc., be all significantly better than placebo group for reagent 1,2,7,8,9, illustrate that tested medicine has obvious curative effects and safety to acute respiratory infection.

Experimental example

Following each experimental example proves technique effect of the present invention.

Involved testing according to following formula respectively for reagent thing 1-9 in following experimental example 1-15:

For reagent 1: genipin-1-β-D gentiobioside with cape jasmine;

For reagent 2: genipin-1-β-D gentiobioside with cape jasmine mixes with weight ratio 1:1 with geniposide;

For reagent 3: genipin-1-β-D gentiobioside with cape jasmine mixes with weight ratio 3:1 with geniposide;

For reagent 4: genipin-1-β-D gentiobioside with cape jasmine mixes with weight ratio 6:1 with geniposide;

For reagent 5: genipin-1-β-D gentiobioside with cape jasmine mixes with weight ratio 1:3 with geniposide;

For reagent 6: genipin-1-β-D gentiobioside with cape jasmine mixes with weight ratio 1:6 with geniposide;

For reagent 7: genipin-1-β-D gentiobioside with cape jasmine, geniposide and shanzhiside methyl ester mix with weight ratio 1:1:0.5;

For reagent 8: extract the Fructus Gardeniae extract containing genipin-1-β-D gentiobioside with cape jasmine and geniposide obtained in embodiment 8;

For reagent 9: extract the Fructus Gardeniae extract containing genipin-1-β-D gentiobioside with cape jasmine and geniposide and shanzhiside methyl ester obtained in embodiment 9.

Experimental example 1: the effect of ICR murine pneumonia model is infected in infected by influenza FM1 strain

Get ICR mice and be divided into 11 groups at random by body weight grade, be respectively Normal group, model control group, 9 confession reagent thing groups, often organize 10.Except Normal group, mice is used ether light anesthesia, infect with 15 LD50 influenza virus liquid (FM1 strain) collunariums, every 30ul.Infect and started administration the same day, press 0.2ml/10g intraperitoneal injection, every day 1 time, continuous 5 days, Normal group and model control group under equal conditions normal saline at every turn.Administration in 5th day is dissected after weighing after 1 hour, claims lung weight, and calculating Lung Exponent and lung index in table 1, and stay specimen for measuring virus load and cytokine in lung tissue.Compare t inspection between result employing group and carry out statistical procedures.

Lung Exponent=lung weight in wet base (g)/body weight (g)

The effect of ICR murine pneumonia model is infected in table 1 infected by influenza FM1 strain

Note: compare ##P<0.01 with normal group; Compare with model control group, * * P<0.01, * P<0.05

Table 1 result shows: after adopting influenza A H1N1 influenza virus FM1 strain virus infecting mouse, mouse lung index obviously increases, and compares have significant difference (P<0.01) with Normal group; Infect after starting to give to treat 5 days for reagent the same day, each group Lung Exponent has obvious reduction, compares have significant difference (P<0.05, P<0.01) with model control group.

Experimental example 2: to the effect of respiratory syncytial virus, parainfluenza virus infection BALB/C mice pulmonary inflammation model

Get Balb/c mice, be divided into 11 groups at random by body weight grade, be respectively Normal group, model control group, 9 confession reagent groups, often organize 10.Except Normal group, mice is used ether light anesthesia, (respiratory syncytial virus is with 10000 LD for the infection of virus liquid collunarium ₅₀, parainfluenza virus is with 1000 LD ₅₀), every 30ul.Infect and started administration the same day, press 0.2ml/10g lumbar injection, every day 1 time, continuous 5 days, Normal group and model control group under equal conditions normal saline at every turn.Dissect after within 5th day, weighing, claim lung weight, calculate Lung Exponent and lung index, in table 2.Compare t inspection between result employing group and carry out statistical procedures.

Lung Exponent=lung weight in wet base (g)/body weight (g);

The effect of table 2 pair viral infection BALB/C mice pulmonary inflammation model

Table 2 result shows: after adopting respiratory syncytial virus and parainfluenza virus infection mice, mouse lung index obviously increases, and compares have significant difference (P<0.01) with Normal group; Infected after starting to give to treat 5 days for reagent the same day, Lung Index of mice infected by Influenza virus all has obvious reducing effect, compares have significant difference (P<0.01, P<0.05) with model control group.

Experimental example 3: the dead protective effect of ICR murine pneumonia model is infected in infected by influenza FM1 strain

Get ICR mice, be divided into 10 groups at random by body weight grade, be respectively model control group, 9 confession reagent groups, often organize 10.Each treated animal ether light anesthesia, with 2 LD ₅₀influenza virus liquid FM1 strain collunarium infects, every 30ul, and then respectively organize mice and press 0.2ml/10g intraperitoneal injection, every day 1 time at every turn, continuous 5 days, model control group is normal saline under equal conditions.Observe the death condition of animal in 2 weeks after infecting, calculate mortality rate, Death prevention rate, Average Survival natural law and increase in life span, the results are shown in Table 3.Compare between result employing group X 2 test and t inspection carry out statistical procedures.

Mortality rate=death toll/animal number × 100%

The dead protective effect of ICR murine pneumonia model is infected in table 3 infected by influenza FM1 strain

Note: compare with model control group, * * P<0.01

Table 3 result shows: adopt in influenza A H1N1 influenza virus FM1 strain virus infecting mouse 2 weeks, model group mortality of animals is 100%, and Average Survival natural law is 5.80 days; Give and supply continuous 5 days of reagent, the death toll of each treated animal obviously reduces, and Average Survival natural law obviously extends, and compares have significant difference (P<0.05) with model control group.

Experimental example 4: the impact of virus load in infected by influenza FM1 strain infecting mouse lung tissue

The animal of experimental animal described in this experimental example and experimental example 1 that the process of animal model is had drawn from.

Design of primers and synthesis: for influenza virus (H1N1) gene order upstream and downstream primer respectively:

5 '-GACCAATCCTGTCACCTCTGAC-3 ' and

-5′-GGGCATTTGGACAAACGTCTACG-3′；

Monitor the use amount of total serum IgE, to eliminate the error that between different sample, application of sample causes for house-keeping gene GAPDH() primer of gene order respectively:

5 '-GGTGAAGGTCGGTGTGAACG-3 ' and

5′-CTCGCTCCTGGAAGATGGTG-3′。

Real-time RT-PCR increases: after the RNA using TRIzol reagent to extract in lung tissue, reverse transcription reaction is carried out respectively with above-mentioned H1N1 and GAPDH primer, reaction volume is 20 μ l, reaction system is containing sample rna 2.5 μ l, reverse transcriptase primer 1 μ l, reverse transcriptase 1 μ l, RNA enzyme inhibitor 1 μ l, reverse transcription reaction liquid 5 μ l.Reaction condition is: 65 DEG C of deactivation 5min, 42 DEG C of reverse transcription 1h.PCR reacts, and reaction system 25 μ l: get cDNA2.5 μ l, each 1 μ l of upstream and downstream primer, PCR reactant liquor 12.5 μ l, remains and supply with the water of DEPC process.95 DEG C of denaturation 5min, 1 circulation; 95 DEG C of degeneration 30sec, 59 DEG C of annealing 30sec, 72 DEG C extend 45sec, totally 40 circulations.Carry out melting curve analysis after reaction terminates, to identify the specificity of PCR primer, the results are shown in Table 4.Sequence Detection System software analysis PCR process is used respectively to detect the Ct(Threshold of cycle of sample) value.

Result computational methods and statistical method: this experiment uses the method for relative quantification, and using GAPDH as internal reference, select a sample (Con) as Calibrator, computational methods are as follows:

Con△Ct＝Con Ct-Con GAPDH Ct；

Sample △ Ct=sample Ct-sample GAPDH Ct;

Sample △ △ Ct=sample △ Ct-Con △ Ct;

2 ^{-Δ Δ Ct}it is the multiple change of the relative Calibrator of each processed group gene expression;

Change=2 of relative amount ^{-Δ Δ Ct}× 100%.

Compare t inspection between result employing group and carry out statistical procedures.

The impact of virus load in table 4 infected by influenza FM1 strain infecting mouse lung tissue

Note: compare with model control group, * P<0.05.

Table 4 result shows: after adopting influenza A H1N1 influenza virus FM1 strain virus infecting mouse, have obvious viral gene expression in lung tissue; Infected after starting to give to treat 5 days for reagent the same day, obviously can reduce virus expression, compare with model control group and have significant difference (P<0.01, P<0.05).

Experimental example 5: to the effect of rabbit herpes simplex keratitis model

Get rabbit 66, be divided into blank group, model group matched group, 9 confession reagent groups at random, often organize 6.After adopting 1% Tetracaine Hydrochloride Solution to carry out topical anesthesia to rabbit right and left eyes, administration group and model control group rabbit aseptic operation sharp knife sheet intersect scuffing right and left eyes corneal epithelium in "+" font, and long is 5.0-6.0mm, and the degree of depth is about 0.5mm.Then draw 40 μ L containing the DMEM drop of HSV-1 virus in right and left eyes anterior corneal surface, passive closed lagophthalmos, massage the eyelid 30 seconds of rabbit gently; The right and left eyes cornea of blank group rabbit scratches the rear 50 μ L of dripping and does not contain viral DMEM.After virus inoculation 24-48h, dyeed by lagophthalmos with 2% fluorescein sodium normal saline solution, observe cornea infection conditions, when point-like, dendroid or map shape focus appear in rabbit corneal, viral infection success is described, animal model makes merit.

Modeling after 48 hours each administration group start eye drop administration, every day 6 times, and within 1,3,5 days, observe pathological changes situation afterwards with administration, the results are shown in Table 5.

Keratopathy (corneal epithelium is painted and substrate is muddy) standards of grading:

0 grade: non-coloring and muddiness;

1 grade: painted and muddy scope is within 25% of cornea area;

2 grades: painted and muddy scope is between the 25%-50% of cornea area;

3 grades: painted and muddy scope is between the 51%-75 of cornea area;

4 grades: painted and muddy scope is more than 75% of cornea area.

The therapeutical effect of table 5 pair rabbit herpes simplex keratitis model

Compare with model control group, * P<0.05, * * P<0.01

Table 5 result shows: in duration of test, all anosis sell of one's property of blank treated animal cornea is raw; After 48 hours, all the other are all positive after respectively organizing fluorescein sodium dyeing in modeling, and cornea lesion degree is balanced; Each for reagent group rabbit cornea pathological changes after administration all can alleviate viral infection on the 3rd day afterwards in various degree, compare with model control group and have significant difference (p<0.05).

Experimental example 6: CoXB3-Nacy strain virus is infected to the impact causing Balb/c mouse core myositis model

220 male Balb/c mices are divided into: Normal group (10), model control group (30), and 9 for reagent group (each group 20).Except Normal group, often organize intraperitoneal inoculation 10 ^-4the CoXB3-Nacy strain virus liquid 0.1ml of dilution.Then respectively organize mice and press 0.2ml/10g intraperitoneal injection at every turn, every day 1 time, continuous 12 days, observe to the 15th day, weigh after record animal dead situation, off-test, get blood centrifugal that myocardial enzymes surveyed by serum, heart is weighed, calculate cardiac index, put in formalin and do heart pathological section, the results are shown in Table 6.

Table 6 infects the impact causing Balb/c mouse core myositis model on CoXB3-Nacy strain virus

Compare with model control group, * P<0.05, * * P<0.01

Table 6 result shows: after CoXB3-Nacy strain virus infects and causes Balb/c mouse core myositis model, and in the increase of model group cardiac index, serum, AST, LDH content obviously increases, and comparing with Normal group has significant difference; Infect after starting to give to treat 12 days for reagent the same day, each treated animal death toll obviously reduces, AST, LDH content obviously reduces in cardiac index, serum, myocardial histopathology degree of injury obviously alleviates, and compares have significant difference (P<0.01, P<0.05) with model control group.

Experimental example 7: impact Epstein-Barr virus transfectional cell being caused to Transplanted Into Nude Mice tumor model

Nude mouse adaptability is raised 1 week, and routine disinfection in sterilizing room, at right axil subcutaneous vaccination nasopharyngeal carcinoma cells CNE-2 Z suspension 0.2ml (about 0.2x10 ⁶individual cell).After inoculated tumour cell, nude mice is sent back to and shrouds, the spirit of routine observation mice, the situation such as diet and defecation.Inoculate about 1 week, visible inoculation position is subcutaneous has tumor nodule to grow, for Transplanted tumor model builds up.After Xenografts in nude mice grows two weeks, get qualified mice (gross tumor volume >100cm ³) be divided into 10 groups at random, be respectively model control group, often organize 6 for 9 for reagent group, each administration group is according to setting dosage intraperitoneal injection, and 0.2ml/kg/d, model control group is intraperitoneal injection of saline under equal conditions.Each group of administration in every 1 day 1 time, continuous 15 times, puts to death nude mouse after within the 16th day, weighing, and takes out tumor tissues, calculates often to organize that average tumor is heavy, inhibition rate of tumor growth, the results are shown in Table 7, compares T inspection and carry out statistical procedures between result employing group.

Table 7 medicine is on the impact of the Nude Mouse Model that CNE-2Z cell causes

Note: compare * P<0.05 with model control group, * * P<0.01.

Table 7 result shows: the Transplanted Into Nude Mice tumor growth that can obviously suppress CNE-2Z cell to cause after 15 days for reagent administration, tumor compares with model control group significant difference.

Experimental example 8: on the impact of duck hepatitis B model

To infect positive duck 60, and be divided into 10 groups by the value equilibrium of DHBV DNA and HBsAg, be respectively model control group, 9 confession reagent groups, often organize 6, age in animal feeding to 2 week starts administration.Administration group respectively oral cavity fills with hello administration, and every day 1 time, matched group is to starch capsule.The experimental drug time is 28 days, and drug withdrawal observes 7 days.Observation index is as follows:

1, serum DHBV DNA changes situation: in medication 7 days, 14 days, 21 days, 28 days, drug withdrawal external jugular vein blood drawing respectively in 7 days, separation of serum is in-20 DEG C of preservations, and each time point serum sample is unified to be checked.Adopt spot hybridization, prepare the unified detection of DHBV DNA probe with the digoxigenin labeled test kit of Roche Holding Ag, carry out diaphragm scanning with Vuego Scan (Brisa-620ST) scanner; Carry out quantitative analysis with the Discovery Series Quantity One software to speckle, speckle value is volume (volume=intensity × mm ²).Statistics adopts spss software paired t-test, the difference of more variant time point administration group and model group;

2, serum DHBsAg changes situation: in medication 7 days, 14 days, 21 days, 28 days, drug withdrawal external jugular vein blood drawing respectively in 7 days, separation of serum is to be checked in-20 DEG C of preservations.Each time point serum is unified to be detected.ELISA express method, enzyme mark reading apparatus (Bio-TEK company) 490nm is adopted to read O.D value.Statistics adopts spss software paired t-test, the difference of more variant time point administration group and model group.

Result shows:

1, after medication, different time points serum DHBV-DNA titre changes situation: during model control group (starch capsule) medication, the prolongation in time of serum DHBV DNA titre presents ascendant trend gradually; After the medication of each confession reagent group, serum DHBV DNA titre keeps a plateau, and from the 3rd week, serum DHBV DNA titre obviously reduced, and compared and had significant difference (P<0.01), the results are shown in Table 8-1 with model group;

2, serum DHBsAgO.D value change situation after the medication of each group: model control group (starch capsule) serum DHBsAgO.D value prolongation in time within 3 weeks presents ascendant trend gradually, and the 4th week starts there is certain falling, but not lower than before administration; DHBsAgO.D value maintenance one plateau after medication is respectively organized for reagent, without significantly rising, and serum DHBV DNA titre decrease to some degree from the 3rd week, compare with model control group and have significant difference (P<0.01), and without rebound significantly within drug withdrawal 1 week, the results are shown in Table 8-2.

Table 8-1 medicine affects unit to DHBV-DNA titre in Duck hepatitis B model serum: volume

Compare with model control group: * * P<0.01*P<0.05

Table 8-2 medicine affects unit to DHBsAgOD value in Duck hepatitis B model serum: OD value

Compare with model control group: * * P<0.01*P<0.05

Experimental example 9: for reagent antiviral effect in vitro

Get the culture plate growing up to cell monolayer, outwell culture fluid, after rinsing cell face 3 times with cell maintenance medium, inoculate different dilution influenza virus liquid respectively, FM1 strain 10 ^-2.5dilution factor, PR8 strain 10 ^-2dilution factor, PIV-1, HSV-1, HSV-2, RSV, CoxB3, CoxB5 are 100TCID50, and Ad3, Ad7 are 10 ^-2dilution factor, rhinovirus are 10 ^-1.5dilution factor, often kind of strain 100 μ L/ hole.Influenza infection mdck cell; PIV-1, HSV-1, HSV-1, HSV-2, RSV, CoxB3, CoxB5 infect Hep-2 cell; Ad3, Ad7, rhinovirus infection Hela cell.Put 37 DEG C of 5%CO ₂after adsorbing 1h in incubator, then add corresponding dilution for reagent medicinal liquid 100 μ L/ hole, establish normal cell controls and virus control simultaneously.Put 37 DEG C of 5%CO ₂cultivate in incubator, observation of cell pathological changes situation under every day inverted microscope, when virus control group cytopathy is ++++time log.

Cytopathy judges by 6 grade standards:

-: Growth of Cells is normal, occurs without pathological changes;

±: cytopathy is less than 10% of whole monolayer;

+: cytopathy accounts for less than 25% of whole cell monolayer;

++: cytopathy accounts for less than 50% of whole cell monolayer;

+++: cytopathy accounts for less than 75% of whole cell monolayer;

++++: cytopathy accounts for more than 75% of whole cell monolayer.

50% inhibition concentration (IC is calculated by Reed-Muench ₅₀) and therapeutic index (TI), TI=TC ₅₀/ IC ₅₀, the results are shown in Table 9.

Table 9 is for reagent antiviral effect in vitro (therapeutic index)

Table 9 result shows: 9 all have obvious inhibitory action to the strain of H1N1 Influenza B virus, PR8 strain, PIV-1, HSV-1, HSV-2, RSV, CoxB3, CoxB5, Ad3, Ad7, rhinovirus in vitro for reagent.

Experimental example 10: extracorporeal bacteria inhibitor test

The preparation of mother solution: the mother solution before experiment, each medicine deionized water being made into 50mg/ml.

After each mother liquid medicine being done 1:2-1:128 times of doubling dilution with broth bouillon during experiment, add in the 96 disposable Tissue Culture Plates in hole, every hole 150ul, then add 10 ⁶the various bacterium liquid of individual bacterium/ml, 15ul/ hole.Establish bacterial controls and PS contrast simultaneously.Put in 37 DEG C of incubators and cultivate 18 hours, observe bacterial growth situation in each hole under inverted microscope, determine the lowest concentration of drug (MIC) without bacterial growth, the results are shown in Table 10.

Table 10 vitro Drug is to the inhibitory action of bacterial growth

One: indicate without bacterial growth+: indicate bacterial growth

Table 10 result shows: after cultivating through 18 hours, bacterial controls Guan Jun has bacterial growth.Each reagent thing that supplies is to the staphylococcus aureus (type strain 26003 of In vitro culture, 331, 360, 361), Staphylococcus albus (26101), staphylococcus epidermidis (type strain, 104, 108), beta hemolytic streptococcus (10, 12), streptococcus pneumoniae (31001, 160, 162, 163) escherichia coli (type strain 44113, 107, 178), bacillus pyocyaneus (10102, 209, 210, 211), Candida albicans (type strain), pneumobacillus (11, 14), micrococcus catarrhalis (29108) growth in vitro all has obvious inhibitory action.

Experimental example 11: antipyretic effect lipopolysaccharide being caused to rabbit fever models

Lipopolysaccharide configures: test front precision balance and take lipopolysaccharide, be configured to 2.5ug/ml/kg dosage with physiological saline solution.

The selection of rabbit: get rabbit and survey basic anus temperature 2 day morning before the test respectively, makes animal adapt to this operation, screens animal simultaneously.

Formal test: morning on the same day was surveyed body temperature in test, selected the animal even group-division of basic anus temperature value between 38.5 ~ 39.5 DEG C, is respectively blank group, model control group 9 for reagent group, often organizes 6.Each administration treated animal presses 2ml/kg intraperitoneal injection 1 time, and blank and model control group give distilled water under square one.Administration after 1 hour except blank group all the other each treated animals press 2ml/kg body weight auricular vein injection lipopolysaccharide modeling, blank group administration physiological saline.Measure the anus temperature of 1h, 2h, 3h, 4h after modeling respectively, the index being Temperature changing with different time anus temperature and basic anus using warming therapy difference, compare t between result employing group and check and carry out statistical procedures.

Table 11 is on the impact (X ± SD, n=6) of rabbit fever models body temperature caused by lipopolysaccharide

Table 10 result shows: duration of test Normal group animal heat remains on fluctuation in normal range; Model group animal heat remains on plateau after modeling; 2nd littlely there is obvious antipyretic effect up to 4 hours to rabbit fever models caused by lipopolysaccharide upon administration for reagent group, compare with model group and have significant difference (P<0.05).

Experimental example 12: antiinflammatory is tested

1, the impact of inflammatory factor in infected by influenza FM1 strain infecting mouse pulmonary inflammation model lung tissue

Measure the lung tissue of Specimen origin in experimental 1 after viral infection.Assay method is undertaken by test kit, the results are shown in Table 12-1.

The impact of peroxide content in table 12-1 infected by influenza FM1 strain infecting mouse lung tissue

Note: Normal group compares #P<0.05; Compare with model control group, * P<0.05, * * P<0.01

The impact of inflammatory factor content in table 12-2 infected by influenza FM1 strain infecting mouse lung tissue

Note: Normal group compares ##P<0.01; Compare with model control group, * P<0.05, * * P<0.01

Table 12-1 and 12-2 result display: adopt influenza A H1N1 influenza virus FM1 strain infecting mouse, infect after starting to give to treat 5 days for reagent the same day, significantly can reduce mouse lung tissue MDA, INF-r and TNF-a content, compare with model control group and have significant difference (P<0.01, P<0.05).

2, glacial acetic acid is caused to the impact of capillary permeability

Get ICR mice, body weight 19 ± 1g, be divided into Normal group, model control group, 9 confession reagent groups at random.Each administration group presses 0.2ml/10g intraperitoneal injection at every turn, every day 1 time, for three days on end, and 1 hour mouse weights after administration in the 3rd day, tail vein injection 0.5% AZO-blue solution 0.1ml/10g.Lumbar injection 0.6% glacial acetic acid solution immediately, 0.2ml/10g.Mice is put to death in dislocation, and intraperitoneal injection of saline, 8ml/ is only.Soft abdomen massage 20 times, extracts peritoneal fluid.The centrifugal 15min of 3000r/mim, Aspirate supernatant, detects absorbance, the results are shown in Table 12-3 under wavelength 630nm.

Table 12-3 causes the impact of mice capillary permeability on glacial acetic acid

Table 12-3 result display: all can reduce glacial acetic acid for reagent group and cause mouse peritoneal capillary permeability, compare with model control group and have significant difference (P<0.01).

Experimental example 13: acute toxicity test in mice

Drug level designs: each test medicine distilled water is mixed with maximum injectable concentration, is respectively:

For examination thing 1:250mg/ml;

For examination thing 2,3,4,5,6:200mg/ml;

For examination thing 7:150mg/ml;

For examination thing 8,9:130mg/ml;

Get mice 160, male and female half and half, raise in the room temperature environment of 20-22 DEG C, 5 cages, freely absorb feedstuff and water.Fasting was divided into 10 groups at random by body weight after 12 hours, and Normal group and 9 confession reagent administration groups, often organize 20, female half and half.Each administration group gives each medicinal liquid of Cmax by 0.2ml/10g body weight lumbar injection, totally 2 times, and 4 hours, interval, matched group gives isopyknic normal saline.To the untoward reaction of observation post administration animal and death condition, continuous 7 days.Calculate total dosage and clinical multiple.

Maximum dosage-feeding (g/kg)=maximum administration concentration × maximum to volume/10g × 100 × administration number of times

Result of the test shows: after each treated animal administration, and appearance activity reduces, slow to irritant reaction, and symptom occurs usually upon administration for 5-15 minute, recovers upon administration within 5 hours; Body weight increases normal.

Body weight change and total dosage data are in table 13.

Table 13 acute toxicity test in mice unit: g

The acute toxicity tests shows, 9 supply reagent all without overt toxicity.

Experimental example 14: rat chronic toxicity test

1 test material

1.1 test medicine: for reagent 1,2,7,8,9.

1.2 experimental animals: healthy Wistar rat, SPF/VAF level, male and female half and half, body weight 130 ± 10g.Tie up experimental animal Technology Co., Ltd. of tonneau China by Beijing to provide.

1.3 test kits: creatinine (CRE), lot number: 20110808; Glutamate pyruvate transaminase (ALT), lot number: 20110829; Glutamate pyruvate transaminase (AST), lot number: 20110824; Potassium (K), lot number: 20110330; Sodium (Na), lot number: 20110729; Beijing Northization safe clinical reagent company limited product.Alkali phosphatase (AKP), lot number: 110461; Total protein (TP), lot number: 110391; Albumin (ALB), lot number: 110461; T-CHOL (CHOL), lot number: 111201; Glucose (GLU), lot number: 110551; Total bilirubin (TBIL), lot number: 110671; Blood urea nitrogen (UREA), lot number: 110801; Gamma-glutamyl based transferase (GGT), lot number: 110591; Triglyceride (TG), lot number: 114731; Creatine kinase (CK), lot number: 110741; Chlorine (Cl), lot number: 110551, Beijing Zhong Shengbei controls biotechnology joint-stock company product.Prothrombin time (PT) measures test kit, lot number: STG20102-48; Activated partial thromboplastin time (APTT) measures test kit, lot number: ST20201-53, Beijing Steellex Scientific Instrument Company's product.

1.4 testing instruments: full-automatic differential hematology analyzer, model: Sysmex XT-2000iv; Platelet aggregation thrombin analyser, model: LG-PABER-1; Body weight electronic balance, Precisa XS4250C Max:420g d=0.01g, Switzerland precisa Products; Semi-automatic biochemical analyzer RT9000 type, Rayto Life and Analytical Sciences Co., Ltd.'s product; Electrocardiograph ECG-6851K type, Japanese NIHON KOHDEN CORPORATION Products.Uroscopy instrument CLINITEK50 type, German Bayer Products.

2 test methods

2.1. the test period: clinical most long-term therapy 7 days, long term toxicity gets its more than 3 times, therefore is decided to be successive administration 1 month, 2 weeks convalescent periods after drug withdrawal.

2.2 dose design

2.2.1 No. 1,2, reagent is supplied: 20g/kg/d, 10g/kg/d, 5g/kg/d;

2.2.2 No. 7, reagent is supplied: 15g/kg/d, 7.5g/kg/d, 3.25g/kg/d

2.2.3 No. 8,9, reagent is supplied: 25g/kg/d, 12.5g/kg/d, 6.25g/kg/d,

2.3 drug solution preparing: test front distilled water and be mixed with 1, No. 2 medicine containing the medicinal liquid of the medicinal liquid of 2.0g/ml, No. 7 medicine 1.5g/ml, 8, No. 9 medicines containing the medicinal liquid of 2.5g/ml, by 1.0ml/100g body weight gastric infusion as heavy dose of group, in, small dose group does 2 times of doubling dilutions, each dosage groups etc. hold the dense gastric infusion such as not, every day 1 time, Normal group gives isometric distilled water.

2.4 test methods: get rat 120, male and female half and half, sub-cage rearing under room temperature environment, freely takes food and take the photograph water, are divided into 16 groups at random by body weight after adaptability raises 3 days, are respectively Normal group, each medicine three dosage groups, often organize 20, male and female half and half.Administration group gastric infusion every day 1 time, continuous 1 month, matched group under equal conditions gave distilled water.Administration dissected animal after 1 month, often organized 10 (female 5, male 5), and all the other animal drug withdrawals are observed, and dissected after 2 weeks.

2.5 observation index

2.5.1 ordinary circumstance: observe the spirit of animal, activity, food-intake, hair, defecation etc. during administration with or without abnormal conditions.

2.5.2 weigh weekly, according to body weights adjustment dosage; Record food-intake.

2.5.3 routine blood test: measure red blood cell count(RBC) (RBC), hemoglobin (HGB), packed cell volume (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC) (MCHC), numeration of leukocyte (WBC), leukocyte differential count (LYM, NEU, MONO, EO, BASO), platelet count (PLT), reticulocyte count (RET) in after difference 1 month upon administration, drug withdrawal 2 weeks.

2.5.4 blood parameters: measure serum alt, AST, CRE, TBIL, TP, ALB, GLU, ALP, CHO, GGT, TRIG, CK, Urea, K, Na, Cl content in after difference 1 month upon administration, drug withdrawal 2 weeks.

2.5.5 respectively at after administration 1 month, within 2 weeks, trace electrocardiogram after drug withdrawal.

2.5.6 important organ index: dissected animal respectively at after administration 1 month, after drug withdrawal 2 weeks and take the heart, liver, spleen, lung, kidney, brain, thymus, adrenal gland, thyroid, stomach, uterus, ovary, testis, epididymis, weight of prostate, calculate organ index.

2.5.7 histopathologic examination: dissected animal respectively at after administration 1 month, after drug withdrawal 2 weeks, win the main organs such as the heart, liver, spleen, lung, kidney, brain, thymus, adrenal gland, Stomach duodenum, uterus and ovary, macroscopy and histopathologic examination.

2.5.8 relevant pathological section, HE dyes, and 10 × 20 amplifications are taken pictures

3 result of the tests

Rat oral gavage gives for reagent 1, No. 2 20g/kg/d, 10g/kg/d, 5g/kg/d; For reagent No. 7 15g/kg/d, 7.5g/kg/d, 3.25g/kg/d and for reagent 8, No. 9 25g/kg/d, 12.5g/kg/d, 6.25g/kg/d, continuous 1 month, continue observation after drug withdrawal 2 weeks, result shows: during administration, the general activity of animal is normal, the mental status is good; Defecation is normal, occurs without vomiting; Hair smoothing, have no obscission; No abnormality seen bleeding tendency.Body weight, food-intake, electrocardio are had no significant effect; The ponderal index of every biochemical indicator, peripheral hemogram and important organ all fluctuates within normal range; Each internal organs macroscopy is showed no obvious pathological change; Pathological study also has no the pathomorphology change caused by medicine; Have no drug-induced toxicity.

Experimental example 15: test medicine treatment acute respiratory infection clinical research

Be that test medicine treatment acute respiratory infection Clinical efficacy and safety are evaluated in contrast with placebo.

1, test drug, dosage regimen

1.1 test drugs:

A group 30 example: for reagent 1, specification: 50mg/ grain (adopting embodiment 12 method to obtain);

B group 30 example: for reagent 2, specification: 100mg/ grain (adopting embodiment 15 method to obtain);

C group 30 example: for reagent 7, specification: 120mg/ grain (adopting embodiment 16 method to obtain);

D group 30 example: for reagent 8, specification: 200mg/ grain (adopting embodiment 24 method to obtain);

E group 30 example: for reagent 9, specification: 250mg/ grain (adopting embodiment 25 method to obtain).

1.2 contrast medicines: F group 30 example: placebo (simulant).

1.3 dosage regimens: each 2, every day 3 times.It within 3 days, is 1 course for the treatment of.

2 observation index

2.1 demographic data and background information

3.1.1 demography index: age, sex, marriage, height, body weight, nationality, occupation

3.1.2 background information: the flu course of disease, merging disease and treatment history thereof etc.

2.2 safeties detect

2.2.1 general health check-up project, comprises body temperature, breathing, heart rate, the rhythm of the heart etc.

2.2.2 routine blood test;

2.2.3 routine urinalysis;

2.2.4 stool routine examination;

2.2.5 liver function (ALT, AST), renal function (Cr, BUN);

2.2.6 electrocardiogram;

Any untoward reaction that 2.2.7 may occur.

2.3 health giving quality Testing index

2.3.1 curative effect index

I main related symptoms, sign (comprising body temperature-oral temperature);

II cooling onset time;

The III antipyretic time.

2.3.2 secondary efficacy index

I minor symptom, sign;

II tongue, pulse condition;

III total white blood cells and related inflammation index.

2.4 diagnosis index

2.4.1 throat swab is cultivated;

2.4.2 chest X-ray.

2.5 observe time point

2.5.1 temperature taking: take medicine within latter 4 hours every 1 hour, measure, record 1 time, 5-24 hour after taking medicine, observes 1 time for every 2 hours.2-3 days every 4 hour records 1 time after taking medicine.Inpatient is measured by nurse, and out-patient is measured voluntarily by experimenter, record.

2.5.2 symptom, sign, picture of the tongue, pulse condition and numeration of leukocyte are in treating fore-and-aft observing, record 1 time.

2.5.3 safety indexes and health giving quality index are in the forward and backward each inspection for the treatment of 1 time.

2.5.4 diagnostic index detects 1 time before only treating.

The regulation of 2.6 time windows: make a house call for after medication 72-96 hour, time window must not more than 1 day.

3 curative effect determinate standards

3.1 curative effect of disease criterion

3.1.1 fully recover: treat within 3 days, temperature recovery is normal, and cold symptoms all disappears.

3.1.2 effective: to treat within 3 days, temperature recovery is normal, most of transference cure of flu.

3.1.3 effective: treat within 3 days, body temperature reduces than before, the cardinal symptom partial disappearance of flu.

3.1.4 invalid: treat body temperature within 3 days and do not fall or raise, the cardinal symptom of flu is without improvement.

3.1.5 total obvious effective rate=(recovery from illness+effective)/total number of cases × 100%

3.1.6 total effective rate=(recovery from illness+effective+effectively)/total number of cases × 100%

3.2 tcm syndrome curative effect determinate standards (judging curative effect by integration method)

3.2.1 therapeutic index (n)=(before treating the rear integration of integration-treatment)/treat front integration × 100%

3.2.2 fully recover: symptom and sign all disappears, n=100%.

3.2.3 effective: clinical symptoms is clearly better, syndrome total mark comparatively treats front minimizing >=70%.

3.2.4 effective: clinical symptom relief, syndrome total mark comparatively treats front minimizing >=30%.

3.2.5 invalid: clinical symptoms is without being clearly better or increasing the weight of, and syndrome total mark reduces < 30% before comparatively treating.

3.2.6 total obvious effective rate=(recovery from illness+effective)/total number of cases × 100%

3.2.7 total effective rate=(recovery from illness+effective+effectively)/total number of cases × 100%

3.3 cooling onset times: to temperature decline 0.5 DEG C (>=0.5) or/and recover normal (<=37.3 DEG C) required time from taking medicine.

The 3.4 antipyretic times: drop to 37.3 DEG C of (<=37.3) required times first to body temperature from taking medicine.

4 statistical results

4.1 flu curative effects

PP analyzes: flu curative effect A, B, C, D, E, F group cure rate be respectively with 48%, 45%, 35.8%, 38%, 40% and 15.0%; Total obvious effective rate is respectively 88.1%, 86%, 76%, 78%, 82% and 32.0%, and total effective rate is respectively 96.2%, 94%, 84%, 88%, 92% and 78.0%.Flu curative effect cure rate, total obvious effective rate, total effective rate A, B, C, D, E group are all better than F group (P<0.05).

ITT analyzes: flu curative effect A, B, C, D, E, F group cure rate are respectively 54.7%, 52.7%, 34.7%, 44.7%, 48.7% and 15.1%; Total obvious effective rate is respectively 88.5%, 84.7%, 64.7%, 74.7%, 78.7% and 42.1%, and total effective rate is respectively 94.9%, 92.7%, 78.7%, 84.7%, 87.7% and 66.5%.Flu curative effect cure rate, total obvious effective rate, total effective rate A, B, C, D, E group are all better than F group (P<0.05).。

4.2 tcm syndrome curative effects

PP analyzes: tcm syndrome curative effect A, B, C, D, E, F group cure rate are respectively 48.0%, 43.0%, 33.0%, 38.0%, 40.0% and 14.0%; Total obvious effective rate is respectively 89.7%, 84.7%, 61.7%, 71.7%, 78.7% and 40.0%, and total effective rate is respectively 96.2%, 94.2%, 76.2%, 84.2%, 86.2% and 76.0%.Tcm syndrome curative effect cure rate, total obvious effective rate, total effective rate A, B, C, D, E group are all better than F group (P<0.05).

ITT analyzes: tcm syndrome curative effect A, B, C, D, E, F group cure rate are respectively 43.1%, 41.1%, 33.1%, 36.1%, 38.1% and 13.4%; Total obvious effective rate is respectively 85.2%, 81.2%, 71.2%, 76.2%, 78.2% and 41.2%, and total effective rate is respectively 97.9%, 94.9%, 84.9%, 88.9%, 90.9% and 74.8%.Tcm syndrome curative effect cure rate, total obvious effective rate, total effective rate A, B, C, D, E group are all better than F group (P<0.05).

4.3 cooling onset times

PP analyzes: treatment rear meta cooling onset time A, B, E group is 3.0h, C, D group is 3.5h, and each group compares with F group (for 5.0h), and cooling onset time statistics has significant difference (P<0.05).

ITT analyzes: after treatment, meta cooling onset time A, B, E group is 4.0h, C, D group be 5.0h, F group is 6.0h, and lower the temperature onset time and F group comparative statistics of each administration group has significant difference (P<0.05).

4.4 antipyretic times

PP analyzes: rear meta antipyretic time A, B, E group for the treatment of is 4.0h, C, D group is 4.5h, and each group compares with F group (for 7.0h), and antipyretic activity statistics has significant difference (P<0.05).

ITT analyzes: rear meta antipyretic time A, B, E group for the treatment of is 4.5h, C, D group is 5.0h, and each group compares with F group (for 8.0h), and antipyretic activity statistics has significant difference (P<0.05).

4.5 safeties: duration of test has no obvious drug-induced untoward reaction.

5 conclusions are shown by double-blind trial result, are all significantly better than placebo group for reagent 1,2,7,8,9 in cure rate, total obvious effective rate, total effective rate etc., illustrate that tested medicine has obvious curative effects and safety to acute respiratory infection.

Detailed description of the invention

Embodiment 1

Get Fructus Gardeniae 500g, chopping, with 4L30%(V/V) alcohol dipping 1 hour, reflux, extract, 2 hours, then with 3L same concentrations alcohol reflux 1 hour, filter, merging filtrate, decompression recycling ethanol (-9Pa, 60 DEG C), obtained extracting solution; By extracting solution in AB-8 macroporous resin loading, with 2 times of column volume water elutions, eluent is in ADS-7 macroporous resin loading, with 2 times of column volumes, concentration for 30%(V/V) ethanol elution, again in ADS-7 macroporous resin loading after eluent decompression recycling ethanol, continue with 3 times of column volumes, concentration as 50%(V/V) ethanol elution, eluent decompression recycling ethanol, continue concentrating under reduced pressure, concentrated solution dehydrated alcohol process, obtains monomer component genipin-1-β-D gentiobioside with cape jasmine.

Embodiment 2

Get Fructus Gardeniae 1000g, chopping, with 8L70%(V/V) alcohol dipping 1 hour, reflux, extract, 2 hours, then with 7L same concentrations ethanol extraction 1 hour, filter, merging filtrate, decompression recycling ethanol (-9Pa, 60 DEG C), obtained extracting solution; By extracting solution in D101 macroporous resin loading, with 3 times of column volume water elutions, eluent is in S-8 macroporous resin loading, with 3 times of column volumes, concentration for 20%(V/V) ethanol elution, again in S-8 macroporous resin loading after eluent decompression recycling ethanol, continue with 2 times of column volumes, concentration as 40%(V/V) ethanol elution, eluent decompression recycling ethanol, continue concentrating under reduced pressure, concentrated solution dehydrated alcohol process, obtains monomer component genipin-1-β-D gentiobioside with cape jasmine.

Embodiment 3

Get Fructus Gardeniae 600g, chopping, with the room temperature water Soakage extraction 6 hours of 5.4L, 4 hours are extracted again with 4.2L water retting, dipping process needs to stir, filter, merging filtrate, obtain extracting solution, by extracting solution in AB-8 macroporous resin loading, with 2 times of column volume water elutions, eluent is in S-8 macroporous resin loading, with 2 times of column volumes, concentration is 20%(V/V) ethanol elution, again in S-8 macroporous resin loading after eluent decompression recycling ethanol, continue with 2 times of column volumes, concentration is 40%(V/V) ethanol elution, eluent decompression recycling ethanol, continue concentrating under reduced pressure, concentrated solution dehydrated alcohol process, obtain monomer component genipin-1-β-D gentiobioside with cape jasmine.

Embodiment 4

Get Fructus Gardeniae 800g, chopping, with 50 DEG C of water rettings of 7.2L, 50 DEG C of insulation water rettings extract 5 hours, again with 5.6L50 DEG C of water extraction 3 hours, dipping process needs to stir, filter, merging filtrate, obtain extracting solution, by extracting solution in AB-8 macroporous resin loading, with 2 times of column volume water elutions, eluent is in AB-8 macroporous resin loading, with 2 times of column volumes, concentration is 30%(V/V) ethanol elution, again in NKA-2 macroporous resin loading after eluent decompression recycling ethanol, continue with 3 times of column volumes, concentration is 50%(V/V) ethanol elution, eluent decompression recycling ethanol, continue concentrating under reduced pressure, concentrated solution acetone treatment, obtain monomer component genipin-1-β-D gentiobioside with cape jasmine.

Embodiment 5

Get Fructus Gardeniae 1000g, chopping, with 9L water room temperature immersion 0.5 hour, decoct (100 DEG C) to extract 1.5 hours, again with 7L water boiling and extraction 1 hour, filter, filtrate merges, obtain extracting solution, by extracting solution in LSA308 macroporous resin loading, with 2 times of column volume water elutions, eluent is in ADS-7 macroporous resin loading, with 2 times of column volumes, concentration is 30%(V/V) ethanol elution, again in ADS-7 macroporous resin loading after eluent decompression recycling ethanol, continue with 3 times of column volumes, concentration is 50%(V/V) ethanol elution, eluent decompression recycling ethanol, continue concentrating under reduced pressure, concentrated solution acetone treatment, obtain monomer component genipin-1-β-D gentiobioside with cape jasmine.

Embodiment 6

Get Fructus Gardeniae 500g, chopping, 50%(V/V with 3.5L) alcohol dipping 1 hour, reflux, extract, 2 hours, again with 3L same concentrations alcohol reflux 1 hour, filter, filtrate merges, decompression recycling ethanol (-9Pa, 60 DEG C), obtain extracting solution, by extracting solution in AB-8 macroporous resin loading, with 3 times of column volume water elutions, eluent is in S-8 macroporous resin loading, with 2 times of column volumes, concentration is 30%(V/V) ethanol elution, again in LSA308 macroporous resin loading after eluent decompression recycling ethanol, continue with 3 times of column volumes, concentration is 60%(V/V) ethanol elution, eluent decompression recycling ethanol, continue concentrating under reduced pressure, concentrated solution methanol process, obtain monomer component geniposide.

Embodiment 7

Get Fructus Gardeniae 800g, chopping, 50%(V/V with 8.8L) alcohol dipping 1 hour, reflux, extract, 2 hours, again with 4.8L same concentrations alcohol reflux 1 hour, filter, filtrate merges, decompression recycling ethanol (-9Pa, 60 DEG C), obtain extracting solution, by extracting solution in ADS-17 macroporous resin loading, with 2 times of column volume water elutions, eluent is in NKA-2 macroporous resin loading, with 2 times of column volumes, concentration is 30%(V/V) ethanol elution, again in NKA-2 macroporous resin loading after eluent decompression recycling ethanol, continue with 3 times of column volumes, concentration is 50%(V/V) ethanol elution, eluent decompression recycling ethanol, continue concentrating under reduced pressure, concentrated solution ethyl acetate process, obtain monomer component shanzhiside methyl ester.Embodiment 8

Get Fructus Gardeniae 600g, chopping, 50%(V/V with 5.4L) alcohol dipping 1 hour, reflux, extract, 2 hours, again with 3.6L same concentrations alcohol reflux 1 hour, filter, filtrate merges, decompression recycling ethanol (-9Pa, 60 DEG C), obtain extracting solution, by extracting solution in NKA-2 macroporous resin loading, with 2 times of column volume water elutions, eluent is in S-8 macroporous resin loading, with 2 times of column volumes, concentration is 30%(V/V) ethanol elution, eluent decompression recycling ethanol, continue concentrating under reduced pressure, concentrated solution dehydrated alcohol process, genipin-1-β-D gentiobioside with cape jasmine must be contained, geniposide two kinds of compositions are main active component.

Embodiment 9

Get Fructus Gardeniae 1000g, chopping, 50%(V/V with 10L) alcohol dipping 1 hour, reflux, extract, 2 hours, again with 6L same concentrations alcohol reflux 1 hour, filter, filtrate merges, decompression recycling ethanol (-9Pa, 60 DEG C), obtain extracting solution, by extracting solution in ADS-17 macroporous resin loading, with 2 times of column volume water elutions, eluent is in LSA308 macroporous resin loading, with 2 times of column volumes, concentration is 30%(V/V) ethanol elution, eluent decompression recycling ethanol, continue concentrating under reduced pressure, concentrated solution dehydrated alcohol process, genipin-1-β-D gentiobioside with cape jasmine must be contained, geniposide and shanzhiside methyl ester three kinds of compositions are main active component.

Embodiment 10

Get Fructus Gardeniae 600g, chopping, with the room temperature water Soakage extraction 6 hours of 4.2L, again with 4.2L room temperature water Soakage extraction 4 hours, dipping process needs to stir, filter, filtrate merges, obtain extracting solution, by extracting solution in ADS-17 macroporous resin loading, with 3 times of column volume water elutions, eluent is in NKA-2 macroporous resin loading, with 2 times of column volumes, concentration is 20%(V/V) ethanol elution, again in NKA-2 macroporous resin loading after eluent decompression recycling ethanol, continue with 3 times of column volumes, concentration is 40%(V/V) ethanol elution, eluent decompression recycling ethanol, continue concentrating under reduced pressure, concentrated solution dehydrated alcohol process, genipin-1-β-D gentiobioside with cape jasmine must be contained, geniposide and shanzhiside methyl ester three kinds of compositions are main active component.

Embodiment 11

Get Fructus Gardeniae 800g, chopping, with 8.8L water room temperature immersion 0.5 hour, decoct (100 DEG C) to extract 1.5 hours, again with 5.6L water boiling and extraction 1 hour, filter, filtrate merges, obtain extracting solution, by extracting solution in LSA308 macroporous resin loading, with 2 times of column volume water elutions, eluent is in S-8 macroporous resin loading, with 3 times of column volumes, concentration is 20%(V/V) ethanol elution, again in NKA-2 macroporous resin loading after eluent decompression recycling ethanol, continue with 3 times of column volumes, concentration is 40%(V/V) ethanol elution, eluent decompression recycling ethanol, continue concentrating under reduced pressure, concentrated solution acetone treatment, genipin-1-β-D gentiobioside with cape jasmine must be contained, geniposide and shanzhiside methyl ester three kinds of compositions are main active component.

Embodiment 12

Get genipin-1-β-D gentiobioside with cape jasmine 50g(and be purchased from Man Site bio tech ltd, Chengdu), pulverize, cross 120 mesh sieves, be about 250g with the starch of drying and mix homogeneously, cross 120 mesh sieves, fully mix, divide and be packed in capsulae vacuus, make 1000 altogether.The application method of described capsule is oral.

Embodiment 13

Get the monomer component that genipin-1-β-D gentiobioside with cape jasmine 25g(adopts embodiment 1 method obtained), lactose 120g, starch 95g, make suitable wet granular with 5% ethyl cellulose ethanol, cross 80 mesh sieves, 60 DEG C of aeration-dryings, with 20 mesh sieve granulate, add Pulvis Talci and are about 6g, magnesium stearate is about 1g, mix homogeneously, tabletting, makes 1000.The application method of described tablet is oral.

Embodiment 14

Get the monomer component that genipin-1-β-D gentiobioside with cape jasmine 50g(adopts embodiment 3 method obtained), add dextrin 950g, with 24 mesh sieve wet granulations, 70 DEG C of aeration-dryings, make 1000g granule.The application method of described granule is warm boiled water.

Embodiment 15

Get genipin-1-β-D gentiobioside with cape jasmine 50g and geniposide 50g(two kinds of monomers are all purchased from Man Site bio tech ltd, Chengdu), mix and pulverize, cross 120 mesh sieves, be about 200g with the starch of drying to mix homogeneously, cross 120 mesh sieves, abundant mixing, divides and is packed in capsulae vacuus, make 1000 altogether.The application method of described capsule is oral.

Embodiment 16

Get genipin-1-β-D gentiobioside with cape jasmine 48g, geniposide 48g and shanzhiside methyl ester 24g(tri-kinds of monomers respectively and be all purchased from Man Site bio tech ltd, Chengdu), pulverize, cross 120 mesh sieves, be about 180g with the starch of drying to mix homogeneously, cross 120 mesh sieves, abundant mixing, divides and is packed in capsulae vacuus, make 1000 altogether.The application method of described capsule is oral.

Embodiment 17

Get genipin-1-β-D gentiobioside with cape jasmine and geniposide combines that (two kinds of monomers are all purchased from Man Site bio tech ltd, Chengdu, weight ratio is 1:3) 5g, mannitol 0.6g, add sterile water for injection in an aseptic environment and be about 900ml, stirring makes abundant dissolving, add sterile water for injection to 1000ml, add 0.2g active carbon, stir about 15 minutes, filter with the G6 sintered filter funnel through sterilizing, be sub-packed in ampoule, after lyophilization, aseptic sealing by fusing, obtains lyophilized injectable powder.The application method of described lyophilized injectable powder is intramuscular injection after 1. dissolving with sterile water for injection; 2. intravenous drip routinely after mixing with 0.9% sodium chloride injection or 5% glucose injection.

Embodiment 18

Get genipin-1-β-D gentiobioside with cape jasmine 8g(and be purchased from Man Site bio tech ltd, Chengdu), mannitol 1g, add sterile water for injection in an aseptic environment and be about 900ml, stirring makes abundant dissolving, add sterile water for injection to 1000ml, filter with the G6 sintered filter funnel through sterilizing, be sub-packed in ampoule, after lyophilization, aseptic sealing by fusing, obtains lyophilized injectable powder.The application method of described lyophilized injectable powder is intramuscular injection after 1. dissolving with sterile water for injection; 2. intravenous drip routinely after mixing with 0.9% sodium chloride injection or 5% glucose injection.

Embodiment 19

Get the monomer component that genipin-1-β-D gentiobioside with cape jasmine 20g(adopts embodiment 4 method obtained), lactose 155g, Icing Sugar 65g, mixing, appropriate with 15% starch slurry, wet granulation (14 mesh sieve), 60 DEG C of aeration-dryings, mix with about 0.6g magnesium stearate, tabletting makes 1000, obtains sublingual tablet.The application method of described sublingual tablet is sublingual administration.

Embodiment 20

Get the monomer component that genipin-1-β-D gentiobioside with cape jasmine 15g(adopts embodiment 5 method obtained), be crushed to micropowder shape, be mixed into pasty state with appropriate oleic acid, add appropriate F11, with blender mixing, fill, sealing-in dose valve device, pressure injection F12, makes 1000 bottles, obtains spray preparation.The application method of described spray preparation is sprayed to oral cavity and throat portion after shaking up.

Embodiment 21

Get 95g to narrow fatty acid ester, heat fused in water-bath, adds 8g genipin-1-β-D gentiobioside with cape jasmine (being purchased from Man Site bio tech ltd, Chengdu), stir, inject the bolt mould scribbling lubricant, open mould after room temperature cooling, make 100 supp anals.The application method of described supp anal is anum administration routinely.

Embodiment 22

Get genipin-1-β-D gentiobioside with cape jasmine, geniposide, (three kinds of monomers adopt embodiment 1,6 and 7 method to obtain respectively in shanzhiside methyl ester three kinds of composition combinations, weight ratio is 0.5:2:0.2) 8g, sodium chloride for injection 7g, add water for injection and be about 800ml, stirring makes abundant dissolving, inject water to 1000ml, add 0.2g active carbon, stir about 15 minutes, filter, pour into neutral ampoule, 100 DEG C of heat sterilizations 30 minutes, obtain aqueous injection.The application method of described aqueous injection is 1. intramuscular injection; 2. intravenous drip routinely after mixing with 0.9% sodium chloride injection or 5% glucose injection.Embodiment 23

Get genipin-1-β-D gentiobioside with cape jasmine, geniposide, (three kinds of monomers are all purchased from Man Site bio tech ltd, Chengdu in shanzhiside methyl ester three kinds of composition combinations, weight ratio is 1:1:0.5) 12g, mannitol 1.2g, add sterile water for injection in an aseptic environment and be about 900ml, stirring makes abundant dissolving, add sterile water for injection to 1000ml, add 0.2g active carbon, stir about 15 minutes, filter with the G6 sintered filter funnel through sterilizing, be sub-packed in ampoule, after lyophilization, aseptic sealing by fusing, obtains lyophilized injectable powder.The application method of described lyophilized injectable powder is intramuscular injection after 1. dissolving with sterile water for injection; 2. intravenous drip routinely after mixing with 0.9% sodium chloride injection or 5% glucose injection.

Embodiment 24

What get that this description embodiment 8 extracts is main active component 200g containing genipin-1-β-D gentiobioside with cape jasmine and geniposide two kinds of compositions, mixs homogeneously with about 100g starch, 70 DEG C of aeration-dryings, pulverize, cross 80 mesh sieves, be filled to capsulae vacuus, make 1000.The application method of described capsule is oral.

Embodiment 25

What get that this description embodiment 9 extracts is main active component 250g containing genipin-1-β-D gentiobioside with cape jasmine, geniposide and shanzhiside methyl ester three kinds of compositions, mixs homogeneously with about 50g starch, 70 DEG C of aeration-dryings, pulverize, cross 80 mesh sieves, be filled to capsulae vacuus, make 1000.The application method of described capsule is oral.

Embodiment 26

What get that this description embodiment 9 extracts is main active component 45g containing genipin-1-β-D gentiobioside with cape jasmine, geniposide and shanzhiside methyl ester three kinds of compositions, lactose 118g, starch 90g, make suitable particulate with 15% ethyl cellulose ethanol, cross 80 mesh sieves, 60 DEG C of aeration-dryings, with 20 mesh sieve granulate, add Pulvis Talci and are about 6g, magnesium stearate is about 0.8g, mix homogeneously, tabletting, makes 1000.The application method of described tablet is oral.

Embodiment 27

What get that this description embodiment 9 extracts is main active component 18g containing genipin-1-β-D gentiobioside with cape jasmine, geniposide and shanzhiside methyl ester three kinds of compositions, mannitol 1.4g, add sterile water for injection in an aseptic environment and be about 900ml, stir and make abundant dissolving, add sterile water for injection to 1000ml, add 0.3g active carbon, stir about 15 minutes, filters with the G6 sintered filter funnel through sterilizing, is sub-packed in ampoule, after lyophilization, aseptic sealing by fusing, obtains lyophilized injectable powder.The application method of described lyophilized injectable powder is intramuscular injection after 1. dissolving with sterile water for injection; 2. intravenous drip routinely after mixing with 0.9% sodium chloride injection or 5% glucose injection.

Obviously, above-described embodiment is only for clearly example being described, and the restriction not to embodiment.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here exhaustive without the need to also giving all embodiments.And thus the apparent change of extending out or variation be still among the protection domain of the invention.

Claims

1. have antiviral, antibacterial, bring down a fever, the medicine of antiinflammatory, antioxidation, it is characterized in that, described medicine with genipin-1-β-D gentiobioside with cape jasmine for active component.

2. medicine according to claim 1, is characterized in that, the active component of described medicine is:

3. medicine according to claim 1 and 2, is characterized in that, the active component of described medicine is:

4., according to the arbitrary described medicine of claim 1-3, it is characterized in that, described medicine makes clinical or pharmaceutically acceptable tablet, capsule, granule, aqueous injection, injectable powder, lyophilized injectable powder, spray, suppository.

5. the arbitrary described medicine of claim 1-4 prepare antiviral, antibacterial, bring down a fever, application in antiinflammatory, anti-oxidation medicine.

6. application according to claim 5, it is characterized in that, described antiviral refers to resisiting influenza virus, against parainfluenza virus, anti respiratory syncytial virus, rhinovirus, anti-adenovirus, anti-hepatitis virus, anti-Coxsackie virus, anti-Echovirus, anti-herpesvirus, anti EB virus and antiretroviral; Describedly antibacterially refer to anti-Staphylococcus aureus, anti-Staphylococcus albus, anti-staphylococcus epidermidis, anti-group B streptococcus, anti-pneumobacillus, anti-streptococcus pneumoniae, anti-micrococcus catarrhalis, Chinese People's Anti-Japanese Military and Political College enterobacteria, anti Bacillus pyocyaneu Flugge, anti-candida albicans; Described antiinflammatory refers to that antiviral property infects and causes pneumonia, myocarditis, keratitis.

7. the application of the arbitrary described medicine of claim 1-4 in preparation treatment acute respiratory infection, influenza, pneumonia, tracheitis, bronchitis, cough, hepatitis B, herpes zoster, simplex keratitis keratitis, viral myocarditis, ebv infection, retroviral infection disease and bacterial infection disease medicine.

8. the application according to claim 5 or 6 or 7, is characterized in that, the route of administration of described medicine comprises acceptable oral administration, drug administration by injection, intravenous drip administration, sublingual administration, spraying suction and rectally clinically.

9. Fructus Gardeniae extract as active component for the preparation of antiviral, antibacterial, bring down a fever, the application of antiinflammatory, anti-oxidation medicine, it is characterized in that, described Fructus Gardeniae extract is prepared by the following method:

Get Fructus Gardeniae 1 weight portion, chopping, with the alcohol dipping 0.5-1 hour of 5-10 parts by volume 30-60%, reflux, extract, 1-2 hour, again with the 30-60% alcohol reflux 0.5-2 hour of 4-8 parts by volume, filter, merging filtrate,-9Pa, 40-80 DEG C of decompression recycling ethanol, obtain extracting solution, by extracting solution in NKA-2 macroporous resin loading, with 1-4 times of column volume water elution, eluent is in S-8 macroporous resin loading, with 1-4 times of column volume, concentration is the ethanol elution of 20-50%, eluent decompression recycling ethanol, continue concentrating under reduced pressure, concentrated solution dehydrated alcohol process, genipin-1-β-D gentiobioside with cape jasmine must be contained, geniposide two kinds of compositions are main extract, or

10. a Fructus Gardeniae extract as active component for the preparation of the application in treatment acute respiratory infection, influenza, pneumonia, tracheitis, bronchitis, cough, hepatitis B, herpes zoster, simplex keratitis keratitis, viral myocarditis, ebv infection, retroviral infection disease and bacterial infection disease medicine, it is characterized in that, described Fructus Gardeniae extract is prepared by the following method: