CN104510747B - A kind of new medicine use of iridoid glycoside - Google Patents

Abstract

The invention belongs to field of medicaments, and in particular to a kind of β D gentiobioside with cape jasmine of iridoid glycoside compound Geniposide 1 and its with the composition that other compositions compatibility is formed as active ingredient for preparing antiviral, antibacterial, bringing down a fever, the new application of anti-inflammatory, anti-oxidation medicine.It can be clinically used for treating ARI, influenza, pneumonia, virus B hepatitis, herpes zoster, simplex keratitis keratitis, vital myocarditis, ebv infection, retroviral infection disease and bacterial infection disease.

Description

A kind of new medicine use of iridoid glycoside

Technical field

The invention belongs to field of medicaments, and in particular to a kind of iridoid glycoside compound -- Geniposide -1- β-D gentiobioses Glycosides and its composition formed with other compositions compatibility are used to prepare antiviral, antibacterial, brought down a fever, in anti-inflammatory, anti-oxidation medicine New application.

Background technology

ARI is not only common clinical, frequently-occurring disease, especially in recent years SARS, crowd's bird flu, H1N1 The appearance of influenza and H7N9 influenzas makes the mankind at any time among pandemic danger, and existing anti-infectives and vaccine can not have Effect is to the continually changing source of infection.Therefore, the medicine for finding effective and safe is the significant problem of 21 century facing mankind.

Virus B hepatitis is in worldwide prevalence, is a worldwide public health problem, have infectiousness it is strong, The features such as route of transmission is complicated, popular wide, the incidence of disease is high.China is the high infection of HBV, the high popular country of hepatitis B.WHO is by State divides chronic viral hepatitis type B hotspot into.《People's Republic of China's health statistics yearbook in 2012》Disclose：28 In the morbidity of kind infection disease notification and death toll statistics, virus hepatitis stands above the 5th, and China is every year for treating B virus Property hepatitis and its trigger the costly of disease reach hundred million yuans of 300-500.

The medicine of existing treatment virus B hepatitis mainly has：Plain interferon α -2b, Lamivudine, polyethylene glycol are done Disturb plain α -2a, adefovirdipivoxil, Entecavir（Entecavir）Tenofovir disoproxil（TDF）Deng.The advantages of interferon is that the course for the treatment of compares Fixed, patient's curative effect higher to transaminase, virus levels are not bery high is relatively good, and E antigen conversion ratios are also higher.It is lacked Point and obvious, it is necessary to inject, and spray interferon and can produce Neuroleptic Leukocytopenia, heating, bone marrow suppression, parainfluenza sample The adverse reactions such as symptom, although there is the fixed course for the treatment of, subset of patients can change into " "small three positive" " from " great three positive ", most of Or it can not turn out cloudy.Nucleotide drug is oral drugs, convenient to take, there is no direct toxic side effect, curative effect ratio Faster, taking medicine will soon be reduced with restrovirus.Its shortcomings that is that the course for the treatment of is long, and the course for the treatment of is not especially to determine, is had People need 2 years, some possibility it is longer, it is necessary to Long-term taking medicine treat.A further disadvantage is that the meeting during Long-term taking medicine The problem of producing resistance.Therefore, the current clinical treating hepatitis B medicine being still badly in need of safely and effectively, with Anti-HBV activity effect Thing.

Epstein-Barr virus in crowd infected with rapid increase trend.Most non-evident symptons after child's infection, or cause light Disease pharyngitis and the infection of the upper respiratory tract.Primary infection occurs for the adolescence, there are about 50% and infectious mononucleosis occurs.Mainly Propagated by saliva, also can be through post-transfusion.Then Epstein-Barr virus infects bone-marrow-derived lymphocyte in pars oralis pharyngis epithelial cell internal breeding, this A little cells largely enter blood circulation and cause systemic infection, and can hide for a long time in human lymphoid organizes.When body is exempted from During epidemic disease hypofunction, it is possible to trigger kinds of tumors.

Treatment about ebv infection, acyclovir (AC) and DHPG (DHPG) can press down EBV duplications, have certain Curative effect.There are two kinds of vaccines to come out at present, one of them be expressed while China is built with gene engineering method EBVgp320 and HBsAg bovine vaccine vaccine, emphasis use is in high risky area of nasopharyngeal carcinoma.Another is purified virus gp320 memebrane protein vaccines, just in Britain Make small-scale inoculation, to observe whether the vaccine can reduce the incidence of disease of infectious mononucleosis.Therefore, EB is treated The medicine of virus infection will have extensive market prospects and clinical value.

Iridoid is the cyclic monoterpenes derivative containing cyclopentane structure unit, is distributed widely in nature, Rubiaceae, The plants such as Oleaceae, Gentianaceae and Scrophulariaceae contain the constituents more.Iridoid has a variety of pharmacological activity, such as Hypotensive, hepatic cholagogic, antitumor, anti-inflammatory, protection nerve, treatment diabetes and complication etc..

Geniposide -1- β-D gentiobioside with cape jasmine is iridoid effective constituents contained in the plant such as cape jasmine, the compound Also can be by artificial synthesized acquisition.Notification number be CN1706858 Chinese patent disclose from cape jasmine isolate and purify Geniposide -1- β - The method of D gentiobioside with cape jasmine；Chinese patent CN102000102A discloses Geniposide -1- β-D gentiobioside with cape jasmine and is preparing treatment Application in heart failure disease medicine；《Pharmacology and Clinics of Chinese Materia Medica》Page 39 to page 41 report Geniposide -1- of 2nd phase in 2013 β-D gentiobioside with cape jasmine can improve mental and physical efforts caused by yellow Jackets by improving the contractile function of heart and reducing its preload Exhaustion；《New Chinese medicine and clinical pharmacology》Page 8 to page 10 report of 1st phase in 2009, it is double containing Geniposide -1- β-D rough gentian The Fructus Gardeniae total iridoid glycosides of glucosides can intervene the inflammatory reaction after cerebral hemorrhage, prevent Neuron Apoptosis.

The content of the invention

Therefore, the technical problems to be solved by the invention are to provide a kind of Geniposide -1- β-D gentiobioside with cape jasmine（CAS Number：29307-60-6）And combinations thereof be used to prepare antiviral, antibacterial, bring down a fever, be anti-inflammatory, anti-oxidant, and for treating acute exhale Inhale road infection, influenza, pneumonia, tracheitis, bronchitis, cough, virus B hepatitis, herpes zoster, simplex keratitis angle Film inflammation, vital myocarditis, ebv infection, the new use of retroviral infection disease and bacterial infection disease medicine On the way.

Above-mentioned technical problem is solved by following scheme：

One kind has antiviral, antibacterial, brought down a fever, anti-inflammatory, antioxidation medicine, and the medicine is with Geniposide -1- β-D dragons Courage disaccharide glycosides is active component.

It is used to treat ARI, influenza, pneumonia, tracheitis, bronchitis, cough the invention also discloses one kind Cough, virus B hepatitis, herpes zoster, simplex keratitis keratitis, vital myocarditis, ebv infection, reverse transcription disease The medicine of malicious infectious diseases and bacterial infection disease, the medicine is using Geniposide -1- β-D gentiobioside with cape jasmine as activearm Point.

The activearm of the medicine is grouped into Geniposide -1- β-D gentiobioside with cape jasmine or Geniposide -1- β-D rough gentian is double Glucosides and geniposide（No. CAS：24512-63-8）And/or mountain Cape jasmine glycosides methyl esters（No. CAS：64421-28-9）Mixture.

Preferably, the activearm of the medicine is grouped into：

Geniposide -1- β-D gentiobioside with cape jasmine 1-8 parts by weight, geniposide 1-8 parts by weight；Or

Geniposide -1- β-D gentiobioside with cape jasmine 0.5-6 parts by weight, geniposide 0.5-6 parts by weight, mountain Cape jasmine glycosides methyl esters 0.2-4 Parts by weight.

Likewise, the activearm of the medicine is grouped into：

The parts by weight of Geniposide -1- β-D gentiobioside with cape jasmine 1, the parts by weight of geniposide 1；Or

The parts by weight of Geniposide -1- β-D gentiobioside with cape jasmine 3, the parts by weight of geniposide 1；Or

The parts by weight of Geniposide -1- β-D gentiobioside with cape jasmine 6, the parts by weight of geniposide 1；Or

The parts by weight of Geniposide -1- β-D gentiobioside with cape jasmine 1, the parts by weight of geniposide 3；Or

The parts by weight of Geniposide -1- β-D gentiobioside with cape jasmine 1, the parts by weight of geniposide 6；Or

The parts by weight of Geniposide -1- β-D gentiobioside with cape jasmine 1, the parts by weight of geniposide 1 and the weight of mountain Cape jasmine glycosides methyl esters 0.5 Part.

Clinical or pharmaceutically acceptable tablet, capsule, granule, liquid drugs injection, powder-injection, lyophilized is made in the medicine Powder-injection, spray, suppository.

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

It is described antiviral to refer to resisiting influenza virus, anti-parainfluenza virus, anti respiratory syncytial virus, rhinovirus, anti-gland Virus, anti-hepatitis virus, anti-Coxsackie virus, anti-ECHO virus, anti-herpesvirus, anti EB virus and ARV.

The invention also discloses said medicine to prepare treatment ARI, influenza, pneumonia, tracheitis, branch gas It is Guan Yan, cough, virus B hepatitis, herpes zoster, simplex keratitis keratitis, vital myocarditis, ebv infection, inverse Application in Retroviral infectious diseases and bacterial infection disease medicine.

The method of administration of the medicine includes clinically acceptable oral administration, drug administration by injection, drip-feed administration, tongue Lower administration, spraying suction and rectally.

It is used to prepare antiviral, antibacterial as active component the invention also discloses a kind of capejasmine extract, brings down a fever, resists The scorching, application of anti-oxidation medicine, the capejasmine extract are prepared by the following method：：

The parts by weight of cape jasmine 1 are taken, are shredded, with 5--10 parts by volume 30-60% alcohol dipping 0.5-1 hours, refluxing extraction 1-2 Hour, then extracted 0.5-2 hours with the 30-60% alcohol refluxs of 4-8 parts by volume, filtration, merging filtrate, -9Pa, 40-80 DEG C subtracts Push back receipts ethanol, obtain extract solution, by extract solution in NKA-2 macroreticular resin loadings, with 1-4 times of column volume water elution, eluent in S-8 macroreticular resin loadings, using 1-4 times of column volume, concentration as 20-50% ethanol elution, ethanol is recovered under reduced pressure in eluent, continues It is concentrated under reduced pressure, concentrate is handled with absolute ethyl alcohol, is obtained and is containing two kinds of Geniposide -1- β-D gentiobioside with cape jasmine, geniposide compositions Main extract；Or

The parts by weight of cape jasmine 1 are taken, are shredded, with 5--10 parts by volume 30-60% alcohol dipping 0.5-1 hours, refluxing extraction 1- 2 hours, then extracted 0.5-2 hours with the 30-60% alcohol refluxs of 4-8 parts by volume, filtration, merging filtrate, -9Pa, 40-80 DEG C subtracts Push back receipts ethanol, obtain extract solution, by extract solution in ADS-17 macroreticular resin loadings, with 1-4 times of column volume water elution, eluent in LSA308 macroreticular resin loadings, using 1-4 times of column volume, concentration as 20-50% ethanol elution, ethanol is recovered under reduced pressure in eluent, Continue to be concentrated under reduced pressure, concentrate is handled with absolute ethyl alcohol, is obtained containing Geniposide -1- β-D gentiobioside with cape jasmine, geniposide and mountain Cape jasmine Extract based on three kinds of compositions of glycosides methyl esters；Or

The parts by weight of cape jasmine 1 are taken, are shredded, are stirred Soakage extraction 1-5 hours with the room temperature water of 7-11 parts by volume, then with 5-10 bodies Product part room temperature water extraction 1-3 hours, filtration, merging filtrate, extract solution is obtained, by extract solution in ADS-17 macroreticular resin loadings, with 1-4 times of column volume water elution, eluent in NKA-2 macroreticular resin loadings, using 1-4 times of column volume, concentration as 20-50% ethanol Elution, eluent are recovered under reduced pressure after ethanol then at NKA-2 macroreticular resin loadings, continued using 1-4 times of column volume, concentration 30-70% Ethanol elution, eluent is recovered under reduced pressure ethanol, continues to be concentrated under reduced pressure, and concentrate is handled with absolute ethyl alcohol, obtain containing Geniposide- Extract based on three kinds of 1- β-D gentiobioside with cape jasmine, geniposide and mountain Cape jasmine glycosides methyl esters compositions；Or

The parts by weight of cape jasmine 1 are taken, are shredded, with the water room temperature immersion 0.5-1 hours of 7-11 parts by volume, decoct extraction 1-3 hours, Boiled 0.5-2 hours with 5-10 parts by volume decoctings again, filter, merging filtrate, extract solution is obtained, by extract solution in LSA308 macroreticular resins Loading, with 1-3 times of column volume water elution, eluent is in S-8 macroreticular resin loadings, using 1-4 times of column volume, concentration as 20-50%'s Ethanol elution, eluent are recovered under reduced pressure after ethanol then at NKA-2 macroreticular resin loadings, continue using 1-4 times of column volume, concentration as Ethanol is recovered under reduced pressure in 30-70% ethanol elution, eluent, continues to be concentrated under reduced pressure, and concentrate acetone treatment, obtains containing capital Buddhist nun Extract based on three kinds of flat -1- β-D gentiobioside with cape jasmine, geniposide and mountain Cape jasmine glycosides methyl esters compositions；

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

It is used to prepare treatment suddenly as active component the invention also discloses the capejasmine extract that the above method is prepared Property respiratory tract infection, influenza, pneumonia, tracheitis, bronchitis, cough, virus B hepatitis, herpes zoster, herpesviral In property keratitis, vital myocarditis, ebv infection, retroviral infection disease and bacterial infection disease medicine Application.

Medicine of the present invention can by known compound Geniposide -1- β-D gentiobioside with cape jasmine, geniposide and/or Mountain Cape jasmine glycosides methyl esters is directly prepared；Or using capital Buddhist nun is isolated and purified disclosed in Chinese patent CN1706858A from cape jasmine The method of flat -1- β-D gentiobioside with cape jasmine, geniposide and mountain Cape jasmine glycosides methyl esters, and the preparation that is directly used.

The above-mentioned technical proposal of the present invention has advantages below compared with prior art：

1st, the Geniposide -1- β-D gentiobioside with cape jasmine and itself and geniposide and/or the mixture of mountain Cape jasmine glycosides methyl esters, To the strain of H1N1 Influenza B virus infect ICR murine pneumonia models, can reduce infection animal Lung Exponent, reduce mortality of animals, Virus load in lung tissue after extension time-to-live, reduction infection；

2nd, the Geniposide -1- β-D gentiobioside with cape jasmine and itself and geniposide and/or the mixture of mountain Cape jasmine glycosides methyl esters, To parainfluenza virus, respiratory syncytial virus infection BALB/C mice pulmonary inflammation model, the Lung Exponent of infection animal can be reduced；

3rd, the Geniposide -1- β-D gentiobioside with cape jasmine and itself and geniposide and/or the mixture of mountain Cape jasmine glycosides methyl esters, To herpesvirus infection rabbit cornea inflammation model, keratopathy degree can be mitigated；

4th, the Geniposide -1- β-D gentiobioside with cape jasmine and itself and geniposide and/or the mixture of mountain Cape jasmine glycosides methyl esters, ICR mouse core myositis virus models are infected to CoXB3 virus N acy strains, mortality of animals can be reduced, mitigate cardiac index, reduced AST, LDH content in cardiac muscular tissue, and mitigate the pathology damage degree of virus infection rear myocardium tissue；

5th, the Geniposide -1- β-D gentiobioside with cape jasmine and itself and geniposide and/or the mixture of mountain Cape jasmine glycosides methyl esters, To duck hepatitis B virus model, DHBV-DNA and DHBsAg titres in serum can be reduced；

6th, the Geniposide -1- β-D gentiobioside with cape jasmine and itself and geniposide and/or the mixture of mountain Cape jasmine glycosides methyl esters, To CNE-2Z cell Model in Nude Mice, the growth of tumour can be suppressed；

7th, in testing in vitro, the Geniposide -1- β-D gentiobioside with cape jasmine and itself and geniposide and/or mountain Cape jasmine glycosides The mixture of methyl esters, to the strain of H1N1 Influenza B virus, PR8 strains, PIV-1, HSV-1, HSV-2, RSV, CoxB3, CoxB5, Ad3, Ad7, rhinovirus have obvious inhibitory action；

8th, the Geniposide -1- β-D gentiobioside with cape jasmine and itself and geniposide and/or the mixture of mountain Cape jasmine glycosides methyl esters, To the staphylococcus aureus of in vitro culture（Type strain 26003,331,360,361）, staphylococcus albus（26101）, epidermis Staphylococcus（Type strain, 104,108）, beta hemolytic streptococcus（10、12）, streptococcus pneumonia（31001、160、162、 163）Escherichia coli（Type strain 44113,107,178）, Pseudomonas aeruginosa（10102、209、210、211）, Candida albicans（Standard Strain）, pneumobacillus（11、14）, micrococcus catarrhalis（29108）Growth in vitro has obvious inhibitory action；

9th, the Geniposide -1- β-D gentiobioside with cape jasmine and itself and geniposide and/or the mixture of mountain Cape jasmine glycosides methyl esters, There is obvious antipyretic effect to rabbit fever models caused by lipopolysaccharides；

10th, the Geniposide -1- β-D gentiobioside with cape jasmine and itself and geniposide and/or the mixture of mountain Cape jasmine glycosides methyl esters, Mouse lung tissue MDA, INF-r and TNF-a content after influenza infection can be significantly reduced, glacial acetic acid is reduced and causes mouse peritoneal hair Thin vasopermeability, show that there is obvious antiinflammatory action.

11st, after each administration group animal gives each decoction 2 times of Cmax by the intraperitoneal injection of 0.2ml/10g body weight（Interval 4 Hour）, each group animal does not occur death, and body weight increase is normal, shows that 9 tested materials act on without overt toxicity.

12nd, rat is given for reagent 1,2,7,8,9 with clinical 15 times to 70 times of the dosage gavage for intending dosage of people, continuous 4 In week, recover 2 weeks after drug withdrawal, rat oral gavage is given for reagent 1, No. 2 20g/kg/d, 10g/kg/d, 5g/kg/d；For reagent 7 15g/kg/d, 7.5g/kg/d, 3.25g/kg/d and confession reagent 8, No. 9 25g/kg/d, 12.5g/kg/d, 6.25g/kg/d, continuously 1 month, continue observation 2 weeks after drug withdrawal.As a result show：The general activity of animal is normal during administration, the state of mind is good；Size Just normal, no vomiting occurs；Hair smoothing, have no obscission；No abnormality seen hemorrhagic tendency.To body weight, food-intake, electrocardio without Significantly affect；The ponderal index of every biochemical indicator, peripheral hemogram and important organ fluctuates in normal range (NR)；Each internal organs meat Eye examination is showed no obvious pathological change；Pathological study also has no the pathomorphology change caused by medicine； Have no drug-induced toxicity.

13rd, ARI Clinical efficacy and security are treated by control evaluation test medicine of placebo.Give Prescription case is 2 tablets each time, 3 times a day, 3 days is 1 course for the treatment of.Shown by double-blind trial result, controlled for reagent 1,2,7,8,9 More rate, total effective rate, total effective rate etc. are significantly better than placebo, illustrate that institute's reagent thing has to ARI Obvious curative effects and safety.

Experimental example

Following each experimental examples prove technique effect of the present invention.

It is involved in following experimental example 1-15 to be tested respectively according to following formula for reagent thing 1-9：

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

For reagent 2：Geniposide -1- β-D gentiobioside with cape jasmine is with geniposide with weight than 1:1 mixing；

For reagent 3：Geniposide -1- β-D gentiobioside with cape jasmine is with geniposide with weight than 3:1 mixing；

For reagent 4：Geniposide -1- β-D gentiobioside with cape jasmine is with geniposide with weight than 6:1 mixing；

For reagent 5：Geniposide -1- β-D gentiobioside with cape jasmine is with geniposide with weight than 1:3 mixing；

For reagent 6：Geniposide -1- β-D gentiobioside with cape jasmine is with geniposide with weight than 1:6 mixing；

For reagent 7：Geniposide -1- β-D gentiobioside with cape jasmine, geniposide and mountain Cape jasmine glycosides methyl esters are with weight than 1:1:0.5 Mixing；

For reagent 8：The obtained cape jasmine containing Geniposide -1- β-D gentiobioside with cape jasmine and geniposide is extracted in embodiment 8 Extract；

For reagent 9：What extraction obtained in embodiment 9 contains Geniposide -1- β-D gentiobioside with cape jasmine and geniposide and mountain Cape jasmine The capejasmine extract of glycosides methyl esters.

Experimental example 1：The effect of ICR murine pneumonia models is infected in infected by influenza FM1 strains

Take ICR mouse to be randomly divided into 11 groups by body weight grade, respectively Normal group, model control group, 9 supply reagent Thing group, every group 10.In addition to Normal group, by mouse ether light anesthesia, with 15 LD50 influenza virus liquid（FM1 strains） Collunarium infects, every 30ul.The infection same day starts to be administered, and presses 0.2ml/10g intraperitoneal injections, 1 time a day, continuous 5 every time My god, Normal group and model control group under equal conditions physiological saline.Administration in 5th day is dissected after being weighed after 1 hour, claims lung Weight, calculates Lung Exponent and lung index is shown in Table 1, and stays sample to be used to determine virus load and cell factor in lung tissue. As a result use group between compare t examine carry out statistical procedures.

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

The effect of ICR murine pneumonia models is infected in the infected by influenza FM1 strains of table 1

Note：The ##P compared with normal group<0.01；Compared with model control group, * * P<0.01, * P<0.05

The result of table 1 is shown：After H1N1virus FM1 strain virus infecting mouses, mouse Lung Exponent substantially increases Height, relatively there is significant difference with Normal group（P<0.01）；The infection same day starts to give treated 5 days for reagent after, each group Lung Exponent has obvious reduction, relatively has significant difference with model control group（P<0.05、P<0.01）.

Experimental example 2：To Respiratory Syncytial Virus(RSV), the effect of parainfluenza virus infection BALB/C mice pulmonary inflammation model

Balb/c mouse are taken, 11 groups, respectively Normal group, model control group, 9 confessions are randomly divided into by body weight grade Reagent group, every group 10.In addition to Normal group, by mouse ether light anesthesia, the infection of virus liquid collunarium（Respiratory syncystial Virus is with 10000 LD₅₀, parainfluenza virus is with 1000 LD₅₀）, every 30ul.The infection same day starts to be administered, and presses every time 0.2ml/10g is injected intraperitoneally, 1 time a day, continuous 5 days, Normal group and model control group under equal conditions physiological saline. Dissected after weighing within 5th day, claim lung weight, calculated Lung Exponent and lung index, be shown in Table 2.As a result use group between compare t examine into Row statistical procedures.

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

Table 2 infects virus the effect of BALB/C mice pulmonary inflammation model

Note：The ##P compared with normal group<0.01；Compared with model control group, * * P<0.01, * P<0.05

The result of table 2 is shown：After Respiratory Syncytial Virus(RSV) and parainfluenza virus infection mouse, mouse Lung Exponent substantially increases Height, relatively there is significant difference with Normal group（P<0.01）；The infection same day starts to give for after reagent treatment 5 days, infecting Mouse Lung Exponent has obvious reduction to act on, and relatively has significant difference with model control group（P<0.01、P<0.05）.

Experimental example 3：The dead protective effect of ICR murine pneumonia models is infected in infected by influenza FM1 strains

Take ICR mouse, 10 groups be randomly divided into by body weight grade, respectively model control group, 9 supply reagent groups, every group 10 Only.Each group animal ether light anesthesia, with 2 LD₅₀Influenza virus liquid FM1 strains collunarium infects, and every 30ul, then each group is small Mouse presses 0.2ml/10g intraperitoneal injections every time, 1 time a day, continuous 5 days, model control group under equal conditions physiological saline. The death condition of animal in 2 weeks after infecting is observed, calculates the death rate, Death prevention rate, average Survival number of days and increase in life span, It the results are shown in Table 3.As a result use to compare Chi-square Test and t between group and examine and carry out statistical procedures.

The death rate=death toll/animal number × 100%

The dead protective effect of ICR murine pneumonia models is infected in the infected by influenza FM1 strains of table 3

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

The result of table 3 is shown：Using in H1N1virus FM1 strain virus infecting mouse 2 weeks, model group animal dead Rate is 100%, and average Survival number of days are 5.80 days；Give for continuous 5 days of reagent, the death toll of each group animal significantly reduces, and puts down Survival number of days is obviously prolonged, and relatively has significant difference with model control group（P<0.05）.

Experimental example 4：The influence of virus load in infected by influenza FM1 strain infecting mouse lung tissues

Experimental animal described in this experimental example and the processing to animal model have drawn from the animal of experimental example 1.

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

5 '-GACCAATCCTGTCACCTCTGAC-3 ' and

-5′-GGGCATTTGGACAAACGTCTACG-3′；

For house-keeping gene GAPDH（The usage amount of total serum IgE is monitored, to eliminate error caused by sample-adding between different samples）Base Because the primer of sequence is respectively：

5 '-GGTGAAGGTCGGTGTGAACG-3 ' and

5′-CTCGCTCCTGGAAGATGGTG-3′。

Real-time RT-PCR are expanded：After the RNA in lung tissue is extracted using TRIzol reagents, respectively with above-mentioned H1N1 Reverse transcription reaction is carried out with GAPDH primers, reaction volume is 20 μ l, reaction system the μ l containing sample rna 2.5, the μ of reverse transcriptase primer 1 L, the μ l of reverse transcriptase 1, the μ l of RNase inhibitor 1, the μ l of reverse transcription reaction liquid 5.Reaction condition is：65 DEG C of inactivation 5min, 42 DEG C of reverses Record 1h.PCR reacts, the μ l of reaction system 25：CDNA2.5 μ l, each μ l of 1 μ l, PCR reaction solution 12.5 of upstream and downstream primer are taken, residue is used The water of DEPC processing is supplied.95 DEG C of pre-degeneration 5min, 1 circulation；95 DEG C of denaturation 30sec, 59 DEG C of annealing 30sec, 72 DEG C extend 45sec, totally 40 circulations.Reaction carries out melting curve analysis after terminating, and to identify the specificity of PCR primer, the results are shown in Table 4. The Ct of sample is respectively detected using Sequence Detection System software analysis PCR processes（Threshold of cycle）Value.

Result computational method and statistical method：The method that this experiment uses relative quantification, using GAPDH as internal reference, choosing Select a sample（Con）It is as follows as Calibrator, computational methods：

Con △ Ct=Con Ct-Con GAPDH Ct；

Sample △ Ct=sample Ct- sample GAPDH Ct；

Sample △ △ Ct=sample △ Ct-Con △ Ct；

2^-ΔΔCtIt is that multiple of each treatment group gene expression with respect to Calibrator changes；

Change=2 of relative amount^-ΔΔCt×100%。

As a result use group between compare t examine carry out statistical procedures.

The influence of virus load in the infected by influenza FM1 strain infecting mouse lung tissues of table 4

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

The result of table 4 is shown：After H1N1virus FM1 strain virus infecting mouses, have in lung tissue obvious Viral gene expression；The infection same day starts to give for after reagent treatment 5 days, virus expression being can obviously reduce, with model comparison Group, which compares, significant difference（P<0.01、P<0.05）.

Experimental example 5：Effect to rabbit herpes simplex keratitis model

Take rabbit 66, be randomly divided into blank control group, model group control group, 9 supply reagent groups, every group 6.Using 1% After Tetracaine Hydrochloride Solution carries out surface anesthesia to rabbit right and left eyes, administration group and model control group rabbit aseptic operation sharp knife It is in "+" font that piece, which intersects scuffing right and left eyes corneal epithelium, and a length of 5.0-6.0mm, depth is about 0.5mm.Then 40 μ L are drawn The DMEM drops of the virus containing HSV-1 passively close lagophthalmos, gently massage the eyelid of rabbit 30 seconds in right and left eyes anterior corneal surface；It is empty The left and right cornea of white control group rabbit drips DMEMs of the 50 μ L without virus after scratching.After virus inoculation 24-48h, by lagophthalmos with 2% fluorescein sodium normal saline solution dyes, and observes cornea infection conditions, when point-like, dendroid or map shape disease occurs in rabbit corneal During stove, illustrate that virus infects successfully, work(is made in animal model.

Each administration group starts eye drop administration after modeling 48 hours, 6 times a day, and observes lesion feelings after 1,3,5 day with administration Condition, it the results are shown in Table 5.

Keratopathy（Corneal epithelium colours and matrix is muddy）Standards of grading：

0 grade：Non-coloring and muddiness；

1 grade：The scope of coloring and muddiness is within the 25% of cornea area；

2 grades：The scope of coloring and muddiness is between the 25%-50% of cornea area；

3 grades：The scope of coloring and muddiness is between the 51%-75 of cornea area；

4 grades：Coloring and muddy scope are more than the 75% of cornea area.

Therapeutic action of the table 5 to rabbit herpes simplex keratitis model

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

The result of table 5 is shown：Interior blank control group animal corneal disease-free become produces during experiment；After modeling 48 hours remaining It is positive after the dyeing of each group fluorescein sodium, cornea lesion degree is balanced；Respectively for reagent group after administration the 3rd day Mitigate rabbit cornea lesion after virus infects in various degree, relatively have significant difference with model control group（p<0.05）.

Experimental example 6：The influence for causing Balb/c mouse core myositis models is infected CoXB3-Nacy strain virus

220 male Balb/c mouse are divided into：Normal group（10）, model control group（30）, 9 supply reagent group （Each group 20）.In addition to Normal group, every group of intraperitoneal inoculation 10^-4The CoXB3-Nacy strain virus liquid 0.1ml of dilution.Then it is each Group mouse presses 0.2ml/10g intraperitoneal injections every time, 1 time a day, continuous 12 days, observes to the 15th day, records animal dead Weighed after situation, off-test, take blood centrifuge serum surveys myocardial enzymes, heart weighs, and calculates cardiac index, puts formalin In do heart pathological section, the results are shown in Table 6.

Table 6 infects CoXB3-Nacy strain virus the influence for causing Balb/c mouse core myositis models

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

The result of table 6 is shown：After the infection of CoXB3- Nacy strain virus causes Balb/c mouse core myositis models, model group heart AST, LDH content substantially increase in index increase, serum, relatively have significant difference with Normal group；The infection same day starts Give after being treated 12 days for reagent, each group animal dead number significantly reduces, AST, LDH content substantially drop in cardiac index, serum Low, myocardial histopathology degree of injury substantially mitigates, and relatively has significant difference with model control group（P<0.01、P<0.05）.

Experimental example 7：The influence of Transplanted Into Nude Mice tumor model is caused to Epstein-Barr virus transfectional cell

Nude mouse adaptability is raised 1 week, the routine disinfection in desinfection chamber, in right armpit subcutaneous vaccination nasopharyngeal carcinoma cells CNE-2 Z Suspension 0.2ml (about 0.2x10⁶Individual cell).After inoculated tumour cell, nude mice is sent back to and shrouded, the spirit of routine observation mouse, Situations such as diet and defecation.Inoculation 1 week or so, it is seen that inoculation position subcutaneously has tumor nodule growth, is built up for Transplanted tumor model. After Xenografts in nude mice grows two weeks, qualified mouse (gross tumor volume is taken>100cm³) it is randomly divided into 10 groups, respectively model Control group, 9 supply reagent group, and every group 6, each administration group is according to setting dosage intraperitoneal injection, 0.2ml/kg/d, model pair According to group under equal conditions intraperitoneal injection of saline.Each group is administered once for every 1 day, continuous 15 times, is put to death after weighing within the 16th day naked Mouse, tumor tissues are taken out, calculate every group of average knurl weight, inhibition rate of tumor growth, the results are shown in Table 7, as a result using comparing T between group Examine and carry out statistical procedures.

Influence of the medicine of table 7 to Nude Mouse Model caused by CNE-2Z cells

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

The result of table 7 is shown：Transplanted Into Nude Mice tumour caused by can substantially suppressing CNE-2Z cells after being administered 15 days for reagent Growth, tumor relatively have significant difference with model control group.

Experimental example 8：Influence to duck virus B hepatitis model

Positive duck 60 will be infected, is divided into 10 groups, respectively model control group, 9 by DHBV DNA and HBsAg value equilibrium Individual to supply reagent group, every group 6, animal feeding to 2 week old starts to be administered.Oral cavity feeds administration to administration group respectively, one time a day, right Starch capsule is given according to group.The experimental drug time is 28 days, withdrawal observation 7 days.Observation index is as follows：

1st, serum DHBV DNA change situation：In medication 7 days, 14 days, 21 days, 28 days, be discontinued 7 days respectively vena jugularis externa take out Blood, separation serum are unified to be checked in -20 DEG C of preservations, each time point serum sample.Using spot hybridization, with the ground of Roche Holding Ag Gaoxin labelling kit prepares the unified detection of DHBV DNA probes, and film is carried out with Vuego Scan (Brisa-620ST) scanner Piece scans；Quantitative analysis is carried out to spot with the Discovery Series Quantity One softwares, spot value is volume(volume=intensity×mm²).Statistics uses spss software paired t-tests, and more variant time point is given The difference of medicine group and model group；

2nd, serum DHBsAg changes situation：In medication 7 days, 14 days, 21 days, 28 days, be discontinued 7 days respectively vena jugularis externa take out Blood, separation serum are to be checked in -20 DEG C of preservations.Each time point serum is unified to be detected.Using ELISA express methods, enzyme mark reading apparatus （Bio-TEK companies）490nm reads O.D values.Statistics uses spss software paired t-tests, more variant time point administration The difference of group and model group.

As a result show：

1st, different time points serum DHBV-DNA titres change situation after medication：Model control group（Starch capsule）The medication phase Between serum DHBV DNA titres with the time extension present be gradually increasing trend；Respectively for serum DHBV DNA drops after reagent group medication Degree keeps a plateau, and serum DHBV DNA titres substantially reduced since the 3rd week, relatively had conspicuousness poor with model group It is different（P<0.01）, the results are shown in Table 8-1；

2nd, serum DHBsAgO.D value change situations after each group medication：Model control group（Starch capsule）The serum within 3 weeks DHBsAgO.D values present with the extension of time and are gradually increasing trend, begin with certain falling within the 4th week, but before being not below administration；For DHBsAgO.D values keep a plateau after reagent each group medication, rise without obvious, and the serum DHBV since the 3rd week DNA titre decreases to some degree, relatively have significant difference with model control group（P<0.01）, and be discontinued 1 week within nothing Rebound significantly, it the results are shown in Table 8-2.

Influence unit of the table 8-1 medicines to DHBV-DNA titres in Duck hepatitis B model serum：volume

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

Influence unit of the table 8-2 medicines to DHBsAgOD values in Duck hepatitis B model serum：OD values

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

Experimental example 9：For reagent antiviral effect in vitro

The culture plate for having grown up to cell monolayer is taken, outwells nutrient solution, after rinsing cell face 3 times with cell maintenance medium, respectively It is inoculated with the influenza virus liquid of different dilution factors, FM1 strains 10^-2.5Dilution factor, PR8 strains 10^-2Dilution factor, PIV-1, HSV-1, HSV-2, RSV, CoxB3, CoxB5 are 100TCID50, Ad3, Ad7 10^-2Dilution factor, rhinovirus 10^-1.5Dilution factor, every kind of strain 100 μ L/ holes.Influenza infection mdck cell；PIV-1, HSV-1, HSV-1, HSV-2, RSV, CoxB3, CoxB5 infect Hep- 2 cells；Ad3, Ad7, rhinovirus infection Hela cells.Put 37 DEG C of 5%CO₂After adsorbing 1h in incubator, corresponding dilution is added Degree supplies the μ L/ holes of reagent decoction 100, while sets normal cell controls and virus control.Put 37 DEG C of 5%CO₂Cultivated in incubator, Cytopathy situation is observed under daily inverted microscope, when virus control group cytopathy is ++++when log.

Cytopathy is judged by 6 grades of standards：

-：Cell growth is normal, and no lesion occurs；

±：Cytopathy is less than the 10% of whole individual layer；

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

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

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

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

50% inhibition concentration is calculated by Reed-Muench（IC₅₀）And therapeutic index（TI）, TI=TC₅₀/IC₅₀, the results are shown in Table 9。

Table 9 supplies reagent antiviral effect in vitro（Therapeutic index）

The result of table 9 is shown：9 for reagent in vitro to the strain of H1N1 Influenza B virus, PR8 strains, PIV-1, HSV-1, HSV- 2nd, RSV, CoxB3, CoxB5, Ad3, Ad7, rhinovirus have obvious inhibitory action.

Experimental example 10：Extracorporeal bacteria inhibitor test

The preparation of mother liquor：Each medicine is made into 50mg/ml mother liquor before experiment with deionized water.

Each mother liquid medicine is done 1 with broth bouillon during experiment:2-1:After 128 times of doubling dilutions, it is disposable to add to 96 holes In Tissue Culture Plate, per hole 150ul, 10 are added⁶Individual bacterium/ml various bacterium solutions, 15ul/ holes.Set bacterial controls and PS simultaneously Control.Put in 37 DEG C of incubators and cultivate 18 hours, bacterial growth situation in each hole is observed under inverted microscope, it is determined that without bacterial growth Lowest concentration of drug (MIC), the results are shown in Table 10.

Inhibitory action of the vitro Drug of table 10 to bacterial growth

One：Indicate no bacterial growth+：Indicate bacterial growth

The result of table 10 is shown：After culture in 18 hours, bacterial controls Guan Jun has bacterial growth.Respectively for reagent thing to external training Foster staphylococcus aureus（Type strain 26003,331,360,361）, staphylococcus albus（26101）, MRSE （Type strain, 104,108）, beta hemolytic streptococcus（10、12）, streptococcus pneumonia（31001、160、162、163）Large intestine bar Bacterium（Type strain 44113,107,178）, Pseudomonas aeruginosa（10102、209、210、211）, Candida albicans（Type strain）, pneumonia bar Bacterium（11、14）, micrococcus catarrhalis（29108）Growth in vitro has obvious inhibitory action.

Experimental example 11：The antipyretic effect of rabbit fever models is caused to lipopolysaccharides

Lipopolysaccharides configures：Lipopolysaccharides is weighed with precision balance before experiment, 2.5ug/ml/kg is configured to sterile saline Dosage.

The selection of rabbit：Taking rabbit, 2 day morning surveyed basic anus temperature before the test respectively, animal is adapted to this operation, sieves simultaneously Select animal.

Formal test：Experiment morning on the same day surveys body temperature, selects animal of the basic anus temperature value between 38.5~39.5 DEG C uniform 9 packet, respectively blank control group, model control group confession reagent groups, every group 6.Each administration group animal presses 2ml/kg abdominal cavities Drug administration by injection 1 time, blank control and model control group give distilled water under square one.Administration removes blank control group after 1 hour Remaining outer each group animal is by the injection lipopolysaccharides modeling of 2ml/kg body weight auricular vein, blank control group administration physiological saline. The anus temperature of 1h, 2h, 3h, 4h after modeling, the finger using the difference of different time anus temperature and basic anus temperature as Temperature changing are determined respectively Mark, as a result using compare between group t examine carry out statistical procedures.

Influence of the table 11 to rabbit fever models body temperature caused by lipopolysaccharides（X ± SD, n=6）

Note：The ##P compared with normal group<0.01；Compared with model control group, * * P<0.01, * P<0.05

The result of table 10 is shown：Normal group animal heat is maintained in normal range (NR) and fluctuated during experiment；Model group is moved Object temperature is maintained at plateau after modeling；For reagent group upon administration the 2nd hour to 4 hours to fever in rabbits caused by lipopolysaccharides Model has obvious antipyretic effect, relatively has significant difference with model group（P<0.05）.

Experimental example 12：Anti-inflammatory is tested

1st, in infected by influenza FM1 strains infecting mouse pulmonary inflammation model lung tissue inflammatory factor influence

Determine Specimen origin metainfective lung tissue of virus in experimental 1.Assay method is carried out by kit, knot Fruit is shown in Table 12-1.

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

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

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

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

Table 12-1 and 12-2 result is shown：Using H1N1virus FM1 strain infecting mouses, the infection same day start to Give after being treated 5 days for reagent, mouse lung tissue MDA, INF-r and TNF-a content can be significantly reduced, relatively had with model control group Significant difference（P<0.01, P<0.05）.

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

ICR mouse are taken, 19 ± 1g of body weight, are randomly divided into Normal group, model control group, 9 confession reagent groups.Each administration Group presses 0.2ml/10g intraperitoneal injections every time, and one time a day, for three days on end, 1 hour mouse is weighed after administration in the 3rd day, and tail is quiet Arteries and veins injects 0.5% evans blue solution 0.1ml/10g.0.6% glacial acetic acid solution, 0.2ml/10g are injected intraperitoneally immediately.Dislocation is put to death Mouse, and intraperitoneal injection of saline, 8ml/ is only.Under soft abdomen massage 20, peritoneal fluid is extracted.3000r/mim is centrifuged 15min, Aspirate supernatant, absorbance is detected under wavelength 630nm, the results are shown in Table 12-3.

Table 12-3 causes the influence of mouse capillary permeability to glacial acetic acid

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

Table 12-3 results are shown：Glacial acetic acid can be reduced for reagent group and causes mouse peritoneal capillary permeability, with model Control group relatively has significant difference（P<0.01）.

Experimental example 13：Acute toxicity test in mice

Drug concentration designs：Each test medicine is configured to maximum injectable concentration with distilled water, is respectively：

For trying thing 1：250mg/ml；

For trying thing 2,3,4,5,6：200mg/ml；

For trying thing 7：150mg/ml；

For trying thing 8,9：130mg/ml；

Take mouse 160, male and female half and half, raise in 20-22 DEG C of room temperature environment, 5 cages, freely absorb feed and Water.10 groups are randomly divided into by body weight after fasting 12 hours, Normal group and 9 supply reagent administration groups, and every group 20, female is each Half.Each administration group is given each decoction of Cmax by the intraperitoneal injection of 0.2ml/10g body weight, totally 2 times, is spaced 4 hours, control group Give isometric physiological saline.Adverse reaction and death condition to observation post administration animal, continuous 7 days.Calculate total dosage With clinical multiple.

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

Result of the test is shown：Each group animal administration after, appearance activity reduce, it is slow to stimulate the reaction, symptom generally to 5-15 minutes occur after medicine, recover upon administration within 5 hours；Body weight increase is normal.

Changes of weight and total dosage data are shown in Table 13.

The acute toxicity test in mice unit of table 13：g

The acute toxicity tests show that 9 for reagent 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 animal：Healthy Wistar rats, SPF/VAF levels, male and female half and half, 130 ± 10g of body weight.It is logical by Beijing dimension Li Hua experimental animals Technology Co., Ltd. provides.

1.3 kit：Creatinine (CRE), lot number：20110808；Glutamic-pyruvic transaminase（ALT）, lot number：20110829；Paddy third Transaminase（AST）, lot number：20110824；Potassium（K）, lot number：20110330；Sodium（Na）, lot number：20110729；Beijing North health Safe clinical reagent Co., Ltd product.Alkaline phosphatase (AKP), lot number：110461；Total protein (TP), lot number：110391；In vain Albumen (ALB), lot number：110461；T-CHOL (CHOL), lot number：111201；Glucose (GLU), lot number：110551；Total courage Red pigment（TBIL）, lot number：110671；Urea nitrogen (UREA), lot number：110801；Gamma-glutamyl based transferase（GGT）, lot number： 110591；Triglycerides（TG）, lot number：114731；Creatine kinase（CK）, lot number：110741；Chlorine（Cl）, lot number：110551, Beijing Zhong Shengbei control biotechnologys joint-stock company product.Prothrombin time（PT）Determine kit, lot number：STG20102-48； Activated partial thromboplastin time（APTT）Determine kit, lot number：ST20201-53, Beijing Steellex Scientific Instrument Company's production Product.

1.4 testing instruments：Full-automatic differential hematology analyzer, model：Sysmex XT-2000iv；Platelet aggregation coagulates Blood factor analyzer, model：LG-PABER-1；Body weight electronic balance, Precisa XS4250C Max:420g d=0.01g, Switzerland's precisa Products；Semi-automatic biochemical analyzer RT9000 types, Rayto Life and Analytical Sciences Co., Ltd.'s production Product；Electrocardiograph ECG-6851K types, 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 takes its more than 3 times, therefore is set to successive administration 1 month, Convalescence 2 weeks after drug withdrawal.

2.2 dose design

2.2.1 reagent 1,2 is supplied：20g/kg/d, 10g/kg/d, 5g/kg/d；

2.2.2 reagent 7 is supplied：15g/kg/d, 7.5g/kg/d, 3.25g/kg/d

2.2.3 reagent 8,9 is supplied：25g/kg/d, 12.5g/kg/d, 6.25g/kg/d,

2.3 drug solution preparing：Before experiment 1, No. 2 decoction of the medicine containing 2.0g/ml, No. 7 medicine 1.5g/ are configured to distilled water Ml decoction, 8, No. 9 decoctions of the medicine containing 2.5g/ml, it is middle or small by 1.0ml/100g body weight gastric infusion as heavy dose of group Dosage group does 2 times of doubling dilutions, each dosage group with etc. the appearance dense gastric infusion such as not, one time a day, Normal group gives isometric(al) Distilled water.

2.4 test method：Take rat 120, male and female half and half, sub-cage rearing under room temperature environment and takes the photograph water at free feeding, fits Answering property by body weight is randomly divided into 16 groups, respectively three Normal group, each medicine dosage groups after raising 3 days, every group 20, female Hero half and half.The daily gastric infusion of administration group 1 time, continuous 1 month, control group under equal conditions gave distilled water.Administration 1 month After dissect animal, every group 10（Female 5, male 5）, remaining animal withdrawal observation, dissected after 2 weeks.

2.5 observation index

2.5.1 ordinary circumstance：The spirit of observation animal, activity, food-intake, hair, stool and urine etc. are no different during administration Reason condition.

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

2.5.3 blood routine：Respectively upon administration 1 month, be discontinued after 2 weeks determine red blood cell count(RBC)（RBC）, hemoglobin （HGB）, packed cell volume（HCT）, MCVU（MCV）, mean corpuscular hemoglobin（MCH）, mean corpuscular Hemoglobin concentration（MCHC）, white blood cell count(WBC)（WBC）, leukocyte differential count（LYM、NEU、MONO、EO、BASO）, blood platelet meter Number（PLT）, reticulocyte count（RET）.

2.5.4 blood parameters：Respectively upon administration 1 month, be discontinued after 2 weeks determine serum alt, AST, CRE, TBIL, TP, ALB, GLU, ALP, CHO, GGT, TRIG, CK, Urea, K, Na, Cl content.

2.5.5 1 month after the administration, be discontinued after trace electrocardiogram within 2 weeks.

2.5.6 important organ index：1 month after the administration, be discontinued after dissect within 2 weeks animal weigh the heart, liver, spleen, lung, Kidney, brain, thymus gland, adrenal gland, thyroid gland, stomach, uterus, ovary, testis, epididymis, weight of prostate, calculate organ index.

2.5.7 histopathologic examination：1 month after the administration, be discontinued after dissect animal within 2 weeks, win the heart, liver, spleen, The main organs such as lung, kidney, brain, thymus gland, adrenal gland, Stomach duodenum, uterus and ovary, visual inspection and Histopathology inspection Look into.

2.5.8 related pathological section, HE dyeing, 10 × 20 amplifications are taken pictures

3 result of the tests

Rat oral gavage is given for reagent 1, No. 2 20g/kg/d, 10g/kg/d, 5g/kg/d；For No. 7 15g/kg/d of reagent, 7.5g/kg/d, 3.25g/kg/d and confession reagent 8, No. 9 25g/kg/d, 12.5g/kg/d, 6.25g/kg/d, continuous 1 month, stop Continue observation 2 weeks after medicine, as a result show：The general activity of animal is normal during administration, the state of mind is good；Stool and urine is normal, Occur without vomiting；Hair smoothing, have no obscission；No abnormality seen hemorrhagic tendency.To body weight, food-intake, electrocardio without obvious shadow Ring；The ponderal index of every biochemical indicator, peripheral hemogram and important organ fluctuates in normal range (NR)；Each internal organs visual inspection It is showed no obvious pathological change；Pathological study also has no the pathomorphology change caused by medicine；Have no by Drug-induced toxicity.

Experimental example 15：Test medicine treats ARI clinical research

ARI Clinical efficacy and security are treated by control evaluation test medicine of placebo.

1st, test drug, dosage regimen

1.1 test drug：

A groups 30：For reagent 1, specification：50mg/ grains（It is made using the method for embodiment 12）；

B groups 30：For reagent 2, specification：100mg/ grains（It is made using the method for embodiment 15）；

C groups 30：For reagent 7, specification：120mg/ grains（It is made using the method for embodiment 16）；

D groups 30：For reagent 8, specification：200mg/ grains（It is made using the method for embodiment 24）；

E groups 30：For reagent 9, specification：250mg/ grains（It is made using the method for embodiment 25）.

1.2 control medicines：F groups 30：Placebo（Simulant）.

1.3 dosage regimen：2 tablets each time, 3 times a day.3 days are 1 course for the treatment of.

2 observation index

2.1 demographic datas 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, merge disease and its treatment history etc.

2.2 safety detection

2.2.1 general physical examination project, including body temperature, breathing, heart rate, rhythm of the heart etc..

2.2.2 blood routine；

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；

2.2.7 any adverse reaction being likely to occur.

2.3 health giving quality Testing index

2.3.1 curative effect index

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

II cools 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 culture；

2.4.2 chest X-ray.

2.5 observation time points

2.5.1 temperature taking：Measurement in every 1 hour, record 1 time in 4 hours after medication, 5-24 hours after medication, every 2 hours Observation 1 time.The 2-3 days every 4 hour records 1 time after medication.Inpatient is measured by nurse, and out-patient is voluntarily surveyed by subject Amount, record.

2.5.2 symptom, sign, tongue picture, pulse condition and white blood cell count(WBC) in treat fore-and-aft observing, record 1 time.

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

2.5.4 detection 1 time before diagnostic index is only treated.

The regulation of 2.6 time windows：72-96 hours carry out interview after medication, and time window must not exceed 1 day.

3 curative effect determinate standards

3.1 curative effect of disease criterion

3.1.1 recovery from illness：Within treatment 3 days, temperature recovery is normal, and cold symptoms all disappear.

It is 3.1.2 effective：Within treatment 3 days, temperature recovery is normal, and most of symptom of flu disappears.

3.1.3 effectively：Within treatment 3 days, body temperature reduces than before, the cardinal symptom partial disappearance of flu.

It is 3.1.4 invalid：Body temperature does not drop or raised within treating 3 days, and the cardinal symptom of flu is without improvement.

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

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

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

3.2.1 therapeutic index（n）=（Integrated before treatment after integration-treatment）/ treat preceding integration × 100%

3.2.2 recovery from illness：Symptom and sign all disappears, n=100%.

It is 3.2.3 effective：Clinical symptoms are clearly better, and syndrome total mark reduces >=70% before relatively treating.

3.2.4 effectively：Clinical symptom relief, syndrome total mark reduce >=30% before relatively treating.

It is 3.2.5 invalid：No significant improvement or aggravated in clinical symptoms, and syndrome total mark reduces ＜ 30% before relatively treating.

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

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

3.3 cooling onset times：To 0.5 DEG C of temperature decline since medication（>=0.5）Or/and recover normal（<=37.3 ℃）Required time.

3.4 antipyretic times：Drop to 37.3 DEG C first to body temperature since medication（<=37.3）Required time.

4 statistical results

4.1 flu curative effects

PP is analyzed：Curative effect A, B, C, D, E, F group cure rate catch a cold respectively and 48%, 45%, 35.8%, 38%, 40% and 15.0%、；Total effective rate is respectively 88.1%, 86%, 76%, 78%, 82% and 32.0%, total effective rate is respectively 96.2%, 94%, 84%th, 88%, 92% and 78.0%.Flu curative effect cure rate, total effective rate, total effective rate A, B, C, D, E group are superior to F groups（P< 0.05）.

ITT is analyzed：Flu curative effect A, B, C, D, E, F group cure rate is respectively 54.7%, 52.7%, 34.7%, 44.7%, 48.7% and 15.1%；Total effective rate is respectively 88.5%, 84.7%, 64.7%, 74.7%, 78.7% and 42.1%, total effective rate point Wei 94.9%, 92.7%, 78.7%, 84.7%, 87.7% and 66.5%.Catch a cold curative effect cure rate, total effective rate, total effective rate A, B, C, D, E group are superior to F groups（P<0.05）..

4.2 tcm syndrome curative effects

PP is analyzed：Tcm syndrome curative effect A, B, C, D, E, F group cure rate is respectively 48.0%, 43.0%, 33.0%, 38.0%, 40.0% and 14.0%；Total effective rate is respectively 89.7%, 84.7%, 61.7%, 71.7%, 78.7% and 40.0%, total effective rate point Wei 96.2%, 94.2%, 76.2%, 84.2%, 86.2% and 76.0%.Tcm syndrome curative effect cure rate, total effective rate, it is total effectively Rate A, B, C, D, E group is superior to F groups（P<0.05）.

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

4.3 cooling onset times

PP is analyzed：Middle position cooling onset time A, B, E group is 3.0h after treatment, and C, D group are 3.5h, each group and F groups (for 5.0h) compare, cooling onset time statistics has significant difference（P<0.05）.

ITT is analyzed：Middle position cooling onset time A, B, E group is 4.0h after treatment, and C, D group are 5.0h, and F groups are 6.0h, respectively There is significant difference administration group cooling onset time with F group comparative statistics（P<0.05）.

4.4 antipyretic times

PP is analyzed：Antipyretic time A, B, E group of middle position is 4.0h after treatment, and C, D group are 4.5h, each group and F groups (for 7.0h) Compare, antipyretic activity statistics has significant difference（P<0.05）.

ITT is analyzed：Antipyretic time A, B, E group of middle position is 4.5h after treatment, and C, D group are 5.0h, each group and F groups (for 8.0h) Compare, antipyretic activity statistics has significant difference（P<0.05）.

4.5 security：Obvious drug-induced adverse reaction is had no during experiment.

5 conclusions are shown by double-blind trial result, for reagent 1,2,7,8,9 in cure rate, total effective rate, total effective rate etc. Aspect is significantly better than placebo, illustrates that institute's reagent thing has obvious curative effects and safety to ARI.

Embodiment

Embodiment 1

Cape jasmine 500g is taken, shreds, uses 4L30%（V/V）Alcohol dipping 1 hour, refluxing extraction 2 hours, then it is identical dense with 3L Spend alcohol reflux to extract 1 hour, filtration, merging filtrate, ethanol is recovered under reduced pressure（- 9Pa, 60 DEG C）, obtain extract solution；By extract solution in AB-8 macroreticular resin loadings, with 2 times of column volume water elutions, eluent is in ADS-7 macroreticular resin loadings, with 2 times of column volumes, concentration For 30%（V/V）Ethanol elution, eluent is recovered under reduced pressure after ethanol then at ADS-7 macroreticular resin loadings, continues with 3 times of cylinders Product, concentration 50%（V/V）Ethanol elution, eluent is recovered under reduced pressure ethanol, continues to be concentrated under reduced pressure, concentrate absolute ethyl alcohol Processing, obtains monomer component Geniposide -1- β-D gentiobioside with cape jasmine.

Embodiment 2

Cape jasmine 1000g is taken, shreds, uses 8L70%（V/V）Alcohol dipping 1 hour, refluxing extraction 2 hours, then it is identical dense with 7L Spend ethanol to extract 1 hour, filtration, merging filtrate, ethanol is recovered under reduced pressure（- 9Pa, 60 DEG C）, obtain extract solution；By extract solution in D101 Macroreticular resin loading, with 3 times of column volume water elutions, eluent is in S-8 macroreticular resin loadings, using 3 times of column volumes, concentration 20% （V/V）Ethanol elution, eluent is recovered under reduced pressure after ethanol then at S-8 macroreticular resin loadings, continues with 2 times of column volumes, concentration For 40%（V/V）Ethanol elution, eluent is recovered under reduced pressure ethanol, continues to be concentrated under reduced pressure, and concentrate is handled with absolute ethyl alcohol, is obtained Monomer component Geniposide -1- β-D gentiobioside with cape jasmine.

Embodiment 3

Cape jasmine 600g is taken, is shredded, is extracted 4 hours with 5.4L room temperature water Soakage extraction 6 hours, then with 4.2L water rettings, Dipping process needs to stir, and filtration, merging filtrate, extract solution is obtained, by extract solution in AB-8 macroreticular resin loadings, with 2 times of column volumes Water elution, eluent is in S-8 macroreticular resin loadings, using 2 times of column volumes, concentration 20%（V/V）Ethanol elution, eluent subtracts Push back receive ethanol after then at S-8 macroreticular resin loadings, continue using 2 times of column volumes, concentration 40%（V/V）Ethanol elution, elution Ethanol is recovered under reduced pressure in liquid, continues to be concentrated under reduced pressure, and concentrate is handled with absolute ethyl alcohol, and it is double to obtain monomer component Geniposide -1- β-D rough gentian Glucosides.

Embodiment 4

Cape jasmine 800g is taken, is shredded, with 7.2L 50 DEG C of water rettings, 50 DEG C of insulation water rettings extract 5 hours, then with 5.6L50 DEG C water extracts 3 hours, and dipping process needs to stir, and filtration, merging filtrate, extract solution is obtained, by extract solution on AB-8 macroreticular resins Sample, with 2 times of column volume water elutions, eluent is in AB-8 macroreticular resin loadings, using 2 times of column volumes, concentration 30%（V/V）Second Alcohol elutes, and eluent is recovered under reduced pressure after ethanol then at NKA-2 macroreticular resin loadings, continued using 3 times of column volumes, concentration 50%（V/ V）Ethanol elution, eluent is recovered under reduced pressure ethanol, continues to be concentrated under reduced pressure, and concentrate acetone treatment, obtains monomer component capital Buddhist nun Flat -1- β-D gentiobioside with cape jasmine.

Embodiment 5

Cape jasmine 1000g is taken, is shredded, with 9L water room temperature immersion 0.5 hour, is decocted（100℃）Extraction 1.5 hours, then with 7L Water boiling and extraction 1 hour, filtration, filtrate merge, and extract solution are obtained, by extract solution in LSA308 macroreticular resin loadings, with 2 times of cylinders Ponding elutes, and eluent is in ADS-7 macroreticular resin loadings, using 2 times of column volumes, concentration 30%（V/V）Ethanol elution, elution Liquid is recovered under reduced pressure after ethanol then at ADS-7 macroreticular resin loadings, is continued using 3 times of column volumes, concentration 50%（V/V）Ethanol wash De-, ethanol is recovered under reduced pressure in eluent, continues to be concentrated under reduced pressure, concentrate acetone treatment, obtains monomer component Geniposide -1- β-D dragons Courage disaccharide glycosides.

Embodiment 6

Cape jasmine 500g is taken, is shredded, with the 50% of 3.5L（V/V）Alcohol dipping 1 hour, refluxing extraction 2 hours, then with 3L phases With concentration ethanol refluxing extraction 1 hour, filtration, filtrate merges, and ethanol is recovered under reduced pressure（- 9Pa, 60 DEG C）, extract solution is obtained, will be extracted Liquid is in AB-8 macroreticular resin loadings, and with 3 times of column volume water elutions, eluent is in S-8 macroreticular resin loadings, with 2 times of column volumes, dense Spend for 30%（V/V）Ethanol elution, eluent is recovered under reduced pressure after ethanol then at LSA308 macroreticular resin loadings, continues with 3 times of posts Volume, concentration 60%（V/V）Ethanol elution, eluent is recovered under reduced pressure ethanol, continues to be concentrated under reduced pressure, and concentrate is with methanol Reason, obtains monomer component geniposide.

Embodiment 7

Cape jasmine 800g is taken, is shredded, with the 50% of 8.8L（V/V）Alcohol dipping 1 hour, refluxing extraction 2 hours, then with 4.8L Same concentrations alcohol reflux extracts 1 hour, and filtration, filtrate merges, and ethanol is recovered under reduced pressure（- 9Pa, 60 DEG C）, extract solution is obtained, will be carried Taking liquid, with 2 times of column volume water elutions, eluent is in NKA-2 macroreticular resin loadings, with 2 times of posts in ADS-17 macroreticular resin loadings Volume, concentration 30%（V/V）Ethanol elution, eluent is recovered under reduced pressure after ethanol then at NKA-2 macroreticular resin loadings, continues Using 3 times of column volumes, concentration 50%（V/V）Ethanol elution, eluent is recovered under reduced pressure ethanol, continues to be concentrated under reduced pressure, and concentrate is used Ethyl acetate processing, obtains monomer component mountain Cape jasmine glycosides methyl esters.Embodiment 8

Cape jasmine 600g is taken, is shredded, with the 50% of 5.4L（V/V）Alcohol dipping 1 hour, refluxing extraction 2 hours, then with 3.6L Same concentrations alcohol reflux extracts 1 hour, and filtration, filtrate merges, and ethanol is recovered under reduced pressure（- 9Pa, 60 DEG C）, extract solution is obtained, will be carried Taking liquid, with 2 times of column volume water elutions, eluent is in S-8 macroreticular resin loadings, with 2 times of cylinders in NKA-2 macroreticular resin loadings Product, concentration 30%（V/V）Ethanol elution, eluent is recovered under reduced pressure ethanol, continues to be concentrated under reduced pressure, concentrate absolute ethyl alcohol Processing, obtain containing the active component based on two kinds of Geniposide -1- β-D gentiobioside with cape jasmine, geniposide compositions.

Embodiment 9

Cape jasmine 1000g is taken, is shredded, with the 50% of 10L（V/V）Alcohol dipping 1 hour, refluxing extraction 2 hours, then with 6L phases With concentration ethanol refluxing extraction 1 hour, filtration, filtrate merges, and ethanol is recovered under reduced pressure（- 9Pa, 60 DEG C）, extract solution is obtained, will be extracted Liquid is in ADS-17 macroreticular resin loadings, and with 2 times of column volume water elutions, eluent is in LSA308 macroreticular resin loadings, with 2 times of cylinders Product, concentration 30%（V/V）Ethanol elution, eluent is recovered under reduced pressure ethanol, continues to be concentrated under reduced pressure, concentrate absolute ethyl alcohol Processing, obtain containing the active component based on three kinds of Geniposide -1- β-D gentiobioside with cape jasmine, geniposide and mountain Cape jasmine glycosides methyl esters compositions.

Embodiment 10

Cape jasmine 600g is taken, is shredded, with 4.2L room temperature water Soakage extraction 6 hours, then it is small with 4.2L room temperature waters Soakage extraction 4 When, dipping process needs to stir, and filtration, filtrate merges, and extract solution is obtained, by extract solution in ADS-17 macroreticular resin loadings, with 3 times of posts Volume water elution, eluent is in NKA-2 macroreticular resin loadings, using 2 times of column volumes, concentration 20%（V/V）Ethanol elution, wash De- liquid is recovered under reduced pressure after ethanol then at NKA-2 macroreticular resin loadings, is continued using 3 times of column volumes, concentration 40%（V/V）Ethanol Elution, eluent are recovered under reduced pressure ethanol, continue to be concentrated under reduced pressure, and concentrate is handled with absolute ethyl alcohol, is obtained containing Geniposide -1- β-D Active component based on three kinds of gentiobioside with cape jasmine, geniposide and mountain Cape jasmine glycosides methyl esters compositions.

Embodiment 11

Cape jasmine 800g is taken, is shredded, with 8.8L water room temperature immersion 0.5 hour, is decocted（100℃）Extraction 1.5 hours, then with 5.6L water boiling and extractions 1 hour, filtration, filtrate merge, and extract solution are obtained, by extract solution in LSA308 macroreticular resin loadings, with 2 times Column volume water elution, eluent is in S-8 macroreticular resin loadings, using 3 times of column volumes, concentration 20%（V/V）Ethanol elution, wash De- liquid is recovered under reduced pressure after ethanol then at NKA-2 macroreticular resin loadings, is continued using 3 times of column volumes, concentration 40%（V/V）Ethanol Elution, eluent are recovered under reduced pressure ethanol, continue to be concentrated under reduced pressure, concentrate acetone treatment, obtain containing Geniposide -1- β-D rough gentian Active component based on three kinds of disaccharide glycosides, geniposide and mountain Cape jasmine glycosides methyl esters compositions.

Embodiment 12

Take Geniposide -1- β-D gentiobioside with cape jasmine 50g（It is purchased from Chengdu Man Site bio tech ltd）, crush, mistake 120 mesh sieves, it is well mixed with dry starch about 250g, crosses 120 mesh sieves, fully mix, packing is inserted in capsulae vacuus, is made altogether 1000.The application method of the capsule is oral.

Embodiment 13

Take Geniposide -1- β-D gentiobioside with cape jasmine 25g（Using monomer component made from the method for embodiment 1）, lactose 120g, Starch 95g, suitable wet granular is made with 5% ethyl cellulose ethanol, crosses 80 mesh sieves, 60 DEG C of aeration-dryings are whole with 20 mesh sieves Grain, talcum powder about 6g, magnesium stearate about 1g are added, be well mixed, tabletting, be made 1000.The application method of the tablet is Orally.

Embodiment 14

Take Geniposide -1- β-D gentiobioside with cape jasmine 50g（Using monomer component made from the method for embodiment 3）, add dextrin 950g, with 24 mesh sieve wet granulations, 70 DEG C of aeration-dryings, 1000g granules are made.The application method of the granule is opened for temperature Mixing in water for oral taking.

Embodiment 15

Take Geniposide -1- β-D gentiobioside with cape jasmine 50g and geniposide 50g（Two kinds of monomers are purchased from Chengdu Man Site biologies Science and Technology Ltd.）, mix and crush, cross 120 mesh sieves, be well mixed with dry starch about 200g, cross 120 mesh sieves, fully Mix, packing is inserted in capsulae vacuus, and 1000 are made altogether.The application method of the capsule is oral.

Embodiment 16

Geniposide -1- β-D gentiobioside with cape jasmine 48g, geniposide 48g and mountain Cape jasmine glycosides methyl esters 24g are taken respectively（Three kinds of monomers It is purchased from Chengdu Man Site bio tech ltd）, crush, cross 120 mesh sieves, be well mixed with dry starch about 180g, 120 mesh sieves are crossed, are fully mixed, packing is inserted in capsulae vacuus, and 1000 are made altogether.The application method of the capsule is oral.

Embodiment 17

Geniposide -1- β-D gentiobioside with cape jasmine is taken to be combined with geniposide（Two kinds of monomers are purchased from Chengdu Man Site biologies section Skill Co., Ltd, weight ratio are 1:3）5g, mannitol 0.6g, in an aseptic environment plus sterile water for injection about 900ml, stirring make Fully dissolving, sterile water for injection is added to add 0.2g activated carbons to 1000ml, stir about 15 minutes, leaked with sterilized G6 incipient fusions Bucket filtering, is sub-packed in ampoule, sterile sealing, produces freeze drying powder injection after freeze-drying.The freeze drying powder injection uses prescription Formula is intramuscular injection after 1. being dissolved with sterile water for injection；2. after being mixed with 0.9% sodium chloride injection or 5% glucose injection Routinely drip-feed.

Embodiment 18

Take Geniposide -1- β-D gentiobioside with cape jasmine 8g（It is purchased from Chengdu Man Site bio tech ltd）, mannitol 1g, In an aseptic environment plus sterile water for injection about 900ml, stirring make fully to dissolve, and add sterile water for injection to 1000ml, with warp The G6 sintered filter funnels filtering of sterilizing, is sub-packed in ampoule, sterile sealing, produces freeze drying powder injection after freeze-drying.It is described lyophilized The application method of powder-injection is intramuscular injection after 1. being dissolved with sterile water for injection；2. with 0.9% sodium chloride injection or 5% grape Routinely drip-feed after sugared parenteral solution mixing.

Embodiment 19

Take Geniposide -1- β-D gentiobioside with cape jasmine 20g（Using monomer component made from the method for embodiment 4）, lactose 155g, Icing Sugar 65g, mixing, wet granulation appropriate with 15% starch slurry（14 mesh sieves）, 60 DEG C of aeration-dryings, mixed with about 0.6g magnesium stearates Close, tabletting is made 1000, produces sublingual tablets.The application method of the sublingual tablets is sublingual administration.

Embodiment 20

Take Geniposide -1- β-D gentiobioside with cape jasmine 15g（Using monomer component made from the method for embodiment 5）, it is crushed to micro mist Shape, pasty state is mixed into appropriate oleic acid, appropriate F11 is added, is mixed with blender, filling, sealing-in dose valve device, pressure injection F12,1000 bottles are made, produces spray preparation.The application method of the spray preparation is to be sprayed to oral cavity and throat after shaking up Portion.

Embodiment 21

The hemizygous fatty acid esters of 95g are taken, fusing is heated in water-bath, add 8g Geniposide -1- β-D gentiobioside with cape jasmine（It is purchased from Chengdu Man Site bio tech ltd）, stir, injection is scribbled in the bolt mould of lubricant, is opened mould after room temperature cooling, is made Into 100 supp anals.The application method of the supp anal is routinely anum administration.

Embodiment 22

Take Geniposide -1- β-D gentiobioside with cape jasmine, geniposide, three kinds of mountain Cape jasmine glycosides methyl esters is into subassembly（Three kinds of monomer difference It is made using embodiment 1,6 and 7 methods, weight ratio is 0.5:2:0.2）8g, sodium chloride for injection 7g, add water for injection about 800ml, stirring make fully to dissolve, and inject water to 1000ml, add 0.2g activated carbons, stir about 15 minutes, filtering, in pouring into Property ampoule, 100 DEG C of heat sterilizations 30 minutes, produces liquid drugs injection.The application method of the liquid drugs injection is 1. intramuscular injection；2. with Routinely drip-feed after 0.9% sodium chloride injection or the mixing of 5% glucose injection.Embodiment 23

Take Geniposide -1- β-D gentiobioside with cape jasmine, geniposide, three kinds of mountain Cape jasmine glycosides methyl esters is into subassembly（Three kinds of monomers are purchased In Chengdu Man Site bio tech ltd, weight ratio is 1:1:0.5）12g, mannitol 1.2g, add nothing in an aseptic environment Bacterium water for injection about 900ml, stirring make fully to dissolve, and add sterile water for injection to add 0.2g activated carbons, stir about 15 to 1000ml Minute, filtered, be sub-packed in ampoule, sterile sealing, produces freeze drying powder injection after freeze-drying with sterilized G6 sintered filter funnels. The application method of the freeze drying powder injection is intramuscular injection after 1. being dissolved with sterile water for injection；2. with 0.9% sodium chloride injection Or 5% glucose injection mixing after routinely drip-feed.

Embodiment 24

Take what this specification embodiment 8 extracted to contain two kinds of compositions of Geniposide -1- β-D gentiobioside with cape jasmine and geniposide Based on active component 200g, be well mixed with about 100g starch, 70 DEG C of aeration-dryings, crush, cross 80 mesh sieves, be filled to hungry area Capsule, it is made 1000.The application method of the capsule is oral.

Embodiment 25

Take what this specification embodiment 9 extracted to contain Geniposide -1- β-D gentiobioside with cape jasmine, geniposide and mountain Cape jasmine glycosides Active component 250g based on three kinds of compositions of methyl esters, is well mixed with about 50g starch, 70 DEG C of aeration-dryings, crushes, and crosses 80 mesh Sieve, is filled to capsulae vacuus, is made 1000.The application method of the capsule is oral.

Embodiment 26

Take what this specification embodiment 9 extracted to contain Geniposide -1- β-D gentiobioside with cape jasmine, geniposide and mountain Cape jasmine glycosides Active component 45g, lactose 118g, starch 90g based on three kinds of compositions of methyl esters, are made suitably with 15% ethyl cellulose ethanol Particle, 80 mesh sieves are crossed, 60 DEG C of aeration-dryings, with 20 mesh sieve whole grains, add talcum powder about 6g, magnesium stearate about 0.8g, mixing is equal It is even, tabletting, it is made 1000.The application method of the tablet is oral.

Embodiment 27

Take what this specification embodiment 9 extracted to contain Geniposide -1- β-D gentiobioside with cape jasmine, geniposide and mountain Cape jasmine glycosides Active component 18g, mannitol 1.4g based on three kinds of compositions of methyl esters, add sterile water for injection about 900ml in an aseptic environment, stir Mixing makes fully to dissolve, and adds sterile water for injection to add 0.3g activated carbons to 1000ml, stir about 15 minutes, hung down with sterilized G6 Molten funnel filtering, is sub-packed in ampoule, sterile sealing, produces freeze drying powder injection after freeze-drying.The use of the freeze drying powder injection Prescription formula is intramuscular injection after 1. being dissolved with sterile water for injection；2. mixed with 0.9% sodium chloride injection or 5% glucose injection Routinely drip-feed after conjunction.

Obviously, above-described embodiment is only intended to clearly illustrate example, and is not the restriction to embodiment.It is right For those of ordinary skill in the art, can also make on the basis of the above description it is other it is various forms of change or Change.There is no necessity and possibility to exhaust all the enbodiments.And the obvious change thus extended out or Among changing still in the protection domain of the invention.

Claims (1)

1. Geniposide -1- β-D gentiobioside with cape jasmine is preparing the pneumonia tool of infected by influenza FM1 strains infection as sole active agent There is the application in the medicine of dead protective effect.