CN104857007A - Novel inhibitor of influenza virus neuraminidase and application thereof - Google Patents
Novel inhibitor of influenza virus neuraminidase and application thereof Download PDFInfo
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- CN104857007A CN104857007A CN201510184295.7A CN201510184295A CN104857007A CN 104857007 A CN104857007 A CN 104857007A CN 201510184295 A CN201510184295 A CN 201510184295A CN 104857007 A CN104857007 A CN 104857007A
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- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
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- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
Abstract
The invention relates to a novel inhibitor of influenza virus neuraminidase, and the inhibitor is specifically icariin and derivatives thereof. The invention also relates to the application of the novel inhibitor to the preparation of medicament, food and health products for the prevention and / or treatment of the human or animal diseases caused by the influenza virus. The diseases comprise but not limited to diseases caused by influenza A virus, influenza B virus, influenza C virus and subtypes, serotypes and gene mutants thereof. By finding the new mechanisms and new functions of icariin and its derivatives, the invention can realize scientific medication, definite object in view and significant effect, and is in favor of human and animal health.
Description
Technical field
The present invention relates to a kind of new inhibitor of influenza neuraminidase, it is specially icariin and derivant thereof.The invention still further relates to this new inhibitor for the preparation of the purposes prevented and/or treated in the medicine of the disease that influenza virus causes, food, health product.Belong to pharmacy and area of pharmacology.
Background technology
Below describe and be beneficial to reader understanding.The information provided or the list of references quoted all can not be considered to prior art of the present invention.
Influenza virus (influenza virus), being called for short influenza virus, is that one can cause the grippal RNA viruses such as people, Canis familiaris L., horse, pig mammal and birds.On taxonomy, influenza virus belongs to orthomyxoviridae family (Orthomyxoviridae), and it can cause acute upper respiratory tract infection, and by air bamboo telegraph, can be very popular by generating period all over the world.This virus was familiar with by British Wilson's Smith (Wilson Smith) in 1933 the earliest, was called H1N1.H represents hemagglutinin; N represents neuraminidase.Digitized representation is dissimilar.Human influenza virus is divided into first (A), second (B), third (C) three type, is the pathogen of influenza (influenza).Wherein influenza A virus antigen easily morphs, and once repeatedly causes worldwide being very popular.
Influenza virus is spherical in shape, and the new strain be separated is then many in thread, and its diameter is between 80 to 120 nanometers, and the length of thread influenza virus can reach 400 nanometers.Influenza structural from outer to inner can be divided into peplos, stromatin and core three part.The core of virus contains the hereditary material of storage Virus Info and copies the necessary enzyme of these information.The hereditary material of influenza virus is sub-thread strand RNA, is abbreviated as ss-RNA, and ss-RNA combines with nucleoprotein (NP), is wound in ribosome (RNP), exists with the form of very high density.Except ribosome, also has the RNA polymerase being responsible for rna transcription.The RNA of influenza A virus is made up of 8 sections.The gene of influenza virus is made up of 8 single stranded RNA fragments, is respectively NA, HA, NP, M, NS, PB1, PB2 and PA gene.They are encoded 10 kinds of albumen: memebrane protein hemagglutinin (Hemagglutinin HA), neuraminic acid (Neuraminidase NA), stromatin (Matrix protein1M1, nucleoprotein (Nucleoprotein NP), 3 kinds of RNA rely on poly-(RNA-dependent RNA polymerase PB1, PB2 and PA) ionophorous protein (Ion channel protein M2) and non-structural protein (Non-structural protein NS1 and NS2).
Stromatin constitutes the outer casing framework of virus, in fact except stromatin (M1), also has memebrane protein (M2) in skeleton.M2 albumen has ion (mainly Na+) passage and regulates the effect of pH value in film, but quantity is little.Stromatin and the outermost peplos of virus are combined closely, and play protection virus core and the effect of maintaining viral space structure.After influenza virus completes its breeding in host cell, stromatin is distributed on host cell membrane inwall, the virus nucleocapsid of molding can position containing stromatin on recognition of host cell membrane, combine with it and form virus structure, and with the form of the sprouting ripe virion of outstanding release (Virion).Peplos is one deck phospholipid bilayer tunic be wrapped in outside stromatin, and this tunic derives from the cell membrane of host, and ripe influenza virus sprouts from host cell, departs from cell, go to infect next target after being wrapped in by the cell membrane of host on one's body.In peplos except phospholipid molecule, also have two kinds of very important glycoproteins: hemagglutinin and neuraminidase.The outstanding virus of this two albuminoid is external, and length is about 10 to 40 nanometers, is referred to as furcella.A general influenza surface can be distributed with 500 hemagglutinin spikes and 100 neuraminidase furcellas.In influenza A virus, the antigen of hemagglutinin and neuraminidase can morph, and this is the foundation distinguishing virus stain hypotype.
Hemagglutinin (HA), in column, can combine with the receptor on the animal erythrocyte surfaces such as people, bird, pig Cavia porcellus and cause blood coagulation, so be referred to as hemagglutinin.Be divided into light chain and heavy chain two parts after hemagglutinin hydrolysis, the latter can combine with the sialic acid receptor on host cell membrane, and the former then can assist peplos and host cell membrane mutually to merge.Hemagglutinin imports in the process of host cell in virus and plays key player.Hemagglutinin has immunogenicity, and antihemagglutinin antibody can neutralized stream Influenza Virus.Neuraminidase (NA) is one is mushroom tetramer glycoprotein, has the sialic activity of hydrolysis.After ripe influenza virus departs from host cell in the mode of sprouting, the hemagglutinin of virus surface can be kept in touch via sialic acid receptor and host cell membrane, need by neuraminidase, sialic acid to be hydrolyzed, cut off the last contact of virus and host cell, virus can be discharged smoothly from host cell, then infect next host cell.Therefore neuraminidase also becomes an action target spot for the treatment of of influenza medicine, and the Oseltamivir (oseltamivir phosphate capsule class) designed for this enzyme is one of foremost Tamiflu.H can be divided into 17 hypotypes (H1 ~ H17), and N has 10 hypotypes (N1 ~ N10).Wherein only H1N1, H2N2, H3N2 main infection mankind, the natural reservoir (of bird flu viruses) of other many hypotype is multiple birds and animal.Wherein maximum to birds harm is H5, H7 and H9 hypotype strain.Generally, bird flu virus can not infect the animal beyond birds and pig.But Hong Kong reported first in 1997 18 routine H5N1 people occurs and infects bird flu case, wherein 6 examples are dead, cause global extensive concern.After 1997, successively there occurs again the event of avian influenza people in the world several times.There is the bird flu viruss such as highly pathogenic H5N1, H7N7, H7N9, H9N2, once morph and there is the transmission capacity of person to person, human avian influenza prevalence can be caused, imply that bird flu virus has had very large potential threat to the mankind.
Because influenza antigen very easily makes a variation, conventional vaccine still can not break out with popular by flu-prevention at present timely and effectively, and therefore anti-influenza virus medicament research is significant in treatment of influenza.The anti-influenza virus medicament applied and studying has diamantane (obsolete) amine drug, influenza virus neuraminidase glycosides enzyme inhibitor, influenza viral receptor blocker and resisiting influenza virus antisense oligonucleotide etc.Amantadine and rimantadine are conventional clinical treatment medicines, but invalid to Type B influenza, easily produce drug resistance and have neurotoxicity; Influenza virus neuraminidase glycosides enzyme inhibitor, if Oseltamivir is current the most effective resisiting influenza virus first-line drug; Influenza viral receptor blocker has obvious antivirus action, the experiment of resisiting influenza virus antisense oligodeoxynucleotidein in vitro proves to suppress virus replication by specificity, likely become influenza specific drug of new generation, but they is still in conceptual phase.
Icariin, Chinese another name is icariine, English Icariin, Icariln, Ieariline by name, English language Chemical name is 3-[(6-Deoxy-alpha-L-mannopyranosyl) oxy]-7-(beta-D-glucopyranosy-loxy)-5-hydroxy-2-(4-methoxyphenyl)-8-(3-methyl-2-buten-1-yl)-4H-1-benzopyran-4-one, and chemical molecular formula is: C
33h
40o
15.Natural icariin is present in Berberidaceae Epimedium (E.pimedium) plant, mainly be distributed in north temperate zone, barrenwort common in the world has 23 kinds, in state-owned 13 kinds, such as: Herba Epimedii (E.brevicornum), Epimedium sagittatum (E.sagitatum), Herba Epimedii (E.icoroanum) etc.Chinese patent application CN1535976A describe in detail the method for extraction purification icariine from barrenwort.Extract from the dry stem and leaf of barrenwort, the icariin after purification is faint yellow acicular crystal powder.Herba Epimedii and its extract icariin, at home for Chinese patent medicine and health product, even the use of more than one thousand years does not find any toxic and side effects for many years, is used for the treatment of acute, chronic hepatitis in Europe, and clinical practice is safe.
Based on the knowledge of inventor, do not report that display icariine influenza neuraminidase activity relationships, particularly icariin and derivant thereof are for the preparation of the purposes preventing and/or treating influenza medicine that influenza virus causes, health product.
Unexpectedly, applicant finds medicine containing icariin and derivant thereof, health product can suppress influenza neuraminidase active effectively, and then reach the value-added effect of drugs of suppression influenza virus, can be used for preventing and/or treating influenza disease, realize object of the present invention.
Summary of the invention
In one aspect, the invention provides a kind of inhibitor of influenza neuraminidase, be specially icariin and derivant thereof.It is for suppressing copying of influenza virus, and then reaches and prevent and/or treat relative influenza disease.
Wherein said icariin and derivant thereof are respectively following formula (I) and the icariin shown in (II) and two derivatives from icariin and investigation thereof, its independent isomer, the raceme of isomer or non-racemic mixture, oxide or its pharmaceutically acceptable salt and hydrate.
Wherein,
Xa, Xb, Xc, Xd, Xe, Xf, Xg, Xh, Xi, X
1, X
2, X
3, X
4, X
5, X
6and X
7independent is oxygen atom (O) or sulphur atom (S);
Alkyl (the C that Ra, Rb, Rc, Rd, Re, Rf, Rg, Rh and Ri are independently hydrogen atom (H), 1-5Ry substitutes
1-C
10) or (C
1-C
10)-O-(C
1-C
10), each alkyl (C
1-C
10) replace with 1-5 R y, each R y is Rq or (C
2-C
10) alkyl, (C
3-C
10) alkyl, (C
3-C
10) cycloalkyl-alkyl, (C
8-C
14) bicyclic alkyl alkyl, (C
8-C
14) tricyclic alkyl alkyl ,-(C
5-C
10) cycloalkyl-alkyl, (C
3-C
10) cycloalkyl, (C
8-C
14) bicyclic alkyl alkyl, (C
8-C
14) tricyclic alkyl alkyl, phenyl, naphthyl, (C
14) aromatic group, they each can replace with one or more Rz, and each Rz is (C
1-C
6) alkyl, (C
2– C
6) thiazolinyl, (C
1– C
6) alkynyl, (C
3– C
8) cycloalkyl, (C
6-C
8) cycloalkenyl group, phenyl, 3-to 5-unit) heterocycle, C (halogen)
3or CH (halogen)
2;
R
1, R
2, R
2, R
4, R
5, R
6, R
7and R
8independent is hydrogen atom (H), or (C
1-C
10) alkyl, (C
2-C
10) thiazolinyl, (C
2-C
10) alkynyl, (C
3-C
10) cycloalkyl, (C
8-C
14) bicyclic alkyl alkyl, (C
8-C
14) tricyclic alkyl alkyl, phenyl, naphthyl, (C
14) aromatic group, they each can be that non-substituted, one or more Rz or Rq replaces, the one or more Rq replacement replaced with more than 1 Rz, and each Rq can be CN, OH, halogen, N
3, NO
2, N (Rz)
2, ═ NR z, CH ═ NRz, NRz OH, ORz, CORz, C (O) ORz, OC (O) Rz, OC (O) ORz, SRz, S (O) Rz or S (O)
2rz; Y
1, Y
2and Y
3independent is CH
3, (C
1-C
12) alkyl, also can with Ry combination replace (C
1-C
12) alkyl, each (C
1-C
12) alkyl can replace with 1-5 R y or an equilibrium ion replaces.
In preferred at one, Xa, Xb, Xb, Xd, Xe, Xf, Xg, Xh, Xi and X
1, X
2, X
3, X
4, X
5, X
6and X
7independent is oxygen atom (O), Ra, Rb, Rc, Rd, Re, Rf, Rg, Rh, Ri, R
1, R
2, R
3, R
4, R
5, R
6, R
7and R
8independent is hydrogen atom (H), Y
1, Y
2and Y
3independent is CH
3.
Another object of the present invention is to provide icariin and derivant thereof for the preparation of the purposes prevented and/or treated in cold medicine that influenza virus causes, health product.Described influenza disease comprises the A type of influenza virus, B-mode and the third type virus and their respective various hypotype thereof and various serotype, various gene mutation body, serotype variant.I.e. H1 ~ H17, the N1 ~ N10 of influenza A virus and various gene mutation bodies, the serotype variant of M1, M2, PA, PB1, PB2, NS1, NS2; B-mode and the third type virus and their respective various hypotype thereof and various serotype, various gene mutation body, serotype variant.
In the world, inventor uses screening from natural Chinese medicinal herb to obtain an influenza virus neuraminidase inhibitor icariin without any side effects and derivant thereof, the disease effectively preventing and/or treat influenza virus to cause first, demonstrates the mechanism of action of this medicine.Accomplish Scientific Usage of Drugs, shoot the arrow at the target.This function of the icariin that has been also Late Cambrian, has filled up Chinese herbal medicine modern domestic, international blank, has been beneficial to Rehabilitation.
Accompanying drawing explanation
Fig. 1, icariin suppress influenza neuraminidase activity analysis.
Fig. 2, icariin suppress influenza virus to copy fluoresent antibody staining analysis at mdck cell.A, be the mdck cell that influenza virus H 5 N 1 infects, do not add icariin, all cells has all infected virus; B, be the mdck cell that influenza virus H 5 N 1 infects, culture medium contains 5 μm of ol icariin, only has a small amount of cell infection virus; C, be the mdck cell that influenza virus H9N2 infects, do not add icariin, all cells has all infected virus; D, be the mdck cell that influenza virus H9N2 infects, culture medium contains 5 μm of ol icariin, only has a small amount of cell infection virus.
Fig. 3, icariin suppress influenza virus to breed flow cytometry analysis at mdck cell.A, be the mdck cell that influenza virus H 5 N 1 infects, do not add icariin, nearly all cell has all infected virus; B, be the mdck cell that influenza virus H 5 N 1 infects, culture medium contains 5 μm of ol icariin, only has a small amount of cell infection virus; C, be the mdck cell that influenza virus H9N2 infects, do not add icariin, nearly all cell has all infected virus; D, be the mdck cell that influenza virus H9N2 infects, culture medium contains 5 μm of ol icariin, only has a small amount of cell infection virus.
Fig. 4, icariin suppress influenza virus to copy real-time n cell function at mdck cell and divide instrument.A, be the mdck cell that influenza virus H 5 N 1 infects, the line with triangle is the result that virus infected cell does not add the cultivation plate hole of icariin; Line with round dot is the control wells of uninfecting virus; Line without round dot and triangle is the plate hole being added with 1 μm of ol of icariin, 5 μm of ol, 10 μm of ol, 20 μm of ol, 50 μm of ol, 100 μm of ol, 200 μm of ol Concentraton gradient.B, be the mdck cell that influenza virus H9N2 infects, the line with triangle is the result that virus infected cell does not add the cultivation plate hole of icariin; Line with round dot is the control wells of uninfecting virus; Line without round dot and triangle is the plate hole being added with 1 μm of ol of icariin, 5 μm of ol, 10 μm of ol, 20 μm of ol, 50 μm of ol, 100 μm of ol, 200 μm of ol Concentraton gradient.
Fig. 5, icariin suppress the zoopery of influenza virus.A, for the average weight increase and decrease situation of each experimental group after mouse inoculation H5N1, the numerical value of initial 10 the mice gross weight ÷ 10 of i.e. 10 mice gross weights-test, solid line represents infecting mouse and uses 30mg/ icariin to prevent ill experimental mice body weight change (experimental group 2); The experimental mice body weight change (experimental group 1) of icariin is not used after represented by dotted arrows mouse infection virus.B is mouse survival rate, and solid line represents infecting mouse and uses 30mg/ icariin to prevent ill experimental mice situation (experimental group 2); The experimental mice Survival (experimental group 1) of icariin is not used after represented by dotted arrows mouse infection virus.
Fig. 6, histopathologic slide analyze.A, normal mouse lung tissue section (experimental group 4).B, normal mouse throws the lung tissue section's (experimental group 3) taking 30mg/ icariin, sees that immunocytes such as having macrophage increases.C, the experimental mice lung tissue section (experimental group 1) of icariin is not used after mouse infection virus, have thrombosis in lungs thin vessels, have a small amount of neutrophil cell degeneration necrosis in broadening alveolar septum, peribronchial has a small amount of lymphocytic infiltration.D, infecting mouse also uses 30mg/ icariin to prevent ill experimental mice lung tissue section (experimental group 2), sees that alveolation is every broadening, with a small amount of macrophages infiltration.
Detailed description of the invention
In order to provide, substance of the present invention be understood, describe some aspect of the present invention, pattern, embodiment, modification and feature with different the level of details hereinafter.
In the practice of the invention, a lot of conventional arts of molecular biology, protein biochemistry, cytobiology, immunology, microbiology and recombinant DNA aspect are employed.These technology are known.Icariin and derivant thereof
In one embodiment, prepare by the icariin defined with following formula (I):
In another embodiment, prepare by the two derivatives from icariin and investigation defined with following formula (II):
Wherein,
Xa, Xb, Xc, Xd, Xe, Xf, Xg, Xh, Xi, X
1, X
2, X
3, X
4, X
5, X
6and X
7independent is oxygen atom (O) or sulphur atom (S);
Alkyl (the C that Ra, Rb, Rc, Rd, Re, Rf, Rg, Rh and Ri are independently hydrogen atom (H), 1-5Ry substitutes
1-C
10) or (C
1-C
10)-O-(C
1-C
10), each alkyl (C
1-C
10) replace with 1-5 R y, each R y is Rq or (C
2-C
10) alkyl, (C
3-C
10) alkyl, (C
3-C
10) cycloalkyl-alkyl, (C
8-C
14) bicyclic alkyl alkyl, (C
8-C
14) tricyclic alkyl alkyl ,-(C
5-C
10) cycloalkyl-alkyl, (C
3-C
10) cycloalkyl, (C
8-C
14) bicyclic alkyl alkyl, (C
8-C
14) tricyclic alkyl alkyl, phenyl, naphthyl, (C
14) aromatic group, they each can replace with one or more Rz, and each Rz is (C
1-C
6) alkyl, (C
2– C
6) thiazolinyl, (C
1– C
6) alkynyl, (C
3– C
8) cycloalkyl, (C
6-C
8) cycloalkenyl group, phenyl, 3-to 5-unit) heterocycle, C (halogen)
3or CH (halogen)
2;
R
1, R
2, R
2, R
4, R
5, R
6, R
7and R
8independent is hydrogen atom (H), or (C
1-C
10) alkyl, (C
2-C
10) thiazolinyl, (C
2-C
10) alkynyl, (C
3-C
10) cycloalkyl, (C
8-C
14) bicyclic alkyl alkyl, (C
8-C
14) tricyclic alkyl alkyl, phenyl, naphthyl, (C
14) aromatic group, they each can be that non-substituted, one or more Rz or Rq replaces, the one or more Rq replacement replaced with more than 1 Rz, and each Rq can be CN, OH, halogen, N
3, NO
2, N (Rz)
2, ═ NR z, CH ═ NRz, NRz OH, ORz, CORz, C (O) ORz, OC (O) Rz, OC (O) ORz, SRz, S (O) Rz or S (O)
2rz; Y
1, Y
2and Y
3independent is CH
3, (C
1-C
12) alkyl, also can with Ry combination replace (C
1-C
12) alkyl, each (C
1-C
12) alkyl can replace with 1-5 R y or an equilibrium ion replaces.
In preferred at one, Xa, Xb, Xb, Xd, Xe, Xf, Xg, Xh, Xi and X
1, X
2, X
3, X
4, X
5, X
6and X
7independent is oxygen atom (O), Ra, Rb, Rc, Rd, Re, Rf, Rg, Rh, Ri, R
1, R
2, R
3, R
4, R
5, R
6, R
7and R
8independent is hydrogen atom (H), Y
1, Y
2and Y
3independent is CH
3.
In a particular implementation, Xa, Xb, Xb, Xd, Xe, Xf, Xg, Xh, Xi and X
1, X
2, X
3, X
4, X
5, X
6and X
7independent is oxygen atom (O), Ra, Rb, Rc, Rd, Re, Rf, Rg, Rh, Ri, R
1, R
2, R
3, R
4, R
5, R
6, R
7and R
8independent is hydrogen atom (H), Y
1, Y
2and Y
3independent is CH
3.
It will be understood by those skilled in the art that described icariin and derivant thereof comprise its independent isomer, the raceme of isomer or non-racemic mixture, oxide or its pharmaceutically acceptable salt and hydrate.
In a specific embodiment of the present invention, by carrying out base group modification to icariin compound, as amination, carboxylated, aldehyde radical, the chemical modification such as Benzylation, activity and the different icariin compound derivatives of physicochemical property can be prepared.
In a detailed description of the invention, the icariin used in the present invention and two derivatives from icariin and investigation thereof can for natural or chemosynthesis with the compound with above-mentioned formula (I) or (II) of modification.Compound can be prepared by known method.Those skilled in the art can easily selective solvent, temperature, pressure and other reaction conditions.Initiation material is commercially available or those skilled in the art can be easy to preparation by known method.
In a preferred embodiment, the natural icariin used in the present invention is Isolation and purification in barrenwort.
Alternatively, natural icariin plant forms from Berberidaceae (Berberidaceae) Epimedium (Epimedium) plant extract purification.Described barrenwort comprises by E. davidi (E.davidii), Baojing Herba Epimedii (E.baojingense), Herba Epimedii Herba Epimedii (E.icoroanum), Hubei Province, river Herba Epimedii (E.fargesii), the western Herba Epimedii in river (E.elongatum), Epimedium acuminatium (E.acuminatum), single leaf Herba Epimedii (E.simplicifolium), short stem Herba Epimedii (E.brachyrrhizum), spend more Herba Epimedii (E.multiflorum), to bestow favour Herba Epimedii (E.enshiense), leather leaf Herba Epimedii (E.reticulatum), light leaf Herba Epimedii (E.sagittatum), wide calyx Herba Epimedii (E.latisepalum), green medicine Herba Epimedii (E.chlorandrum), luxuriant river in Shangdong Province Herba Epimedii (E.platypetalum), Epimedium truncatum (E.truncatum), strong stem Herba Epimedii (E.rhizomatosum), Epimedium (E. sagittatum), spend Herba Epimedii (E.pauciflorum) less, Sky-High Hill Herba Epimedii (E.myrianthum), Epimedium calcaratum (E.ecalcaratum), glandular hair Herba Epimedii (E.glandulosopilosum), lobule Herba Epimedii (E.parvifolium), star flower Herba Epimedii (E.stellulatum), one or more mixture of the group that Zhenping Herba Epimedii (E.ilicifolium) forms.
The definition of some terms used in this description, except as otherwise noted, all technology used herein and scientific words have the meaning equivalent in meaning usually understood with those skilled in the art usually.
The preventive use of icariin and derivant thereof and therapeutic use
In an embodiment of the invention, by using potential candidate compound to carry out enzyme kinetic analysis experiment, it is active that those skilled in the art's screening has suppression influenza neuraminidase, thus suppress influenza virus to be copied.Especially, influenza neuraminidase can be suppressed completely active, for preventing and/or treating flu virus disease by the enzyme kinetic analysis screening experiment determination icariin of candidate compound and derivant thereof.
In the detailed description of the invention of the disease caused various influenza virus, by weight, the proportionate relationship of the interpolation of influenza virus and icariin can be 1:0.00001-10000, is preferably 1:0.0001-1000; Be more preferably 1:0.001-500; Most preferably be 1:0.01-100 and suppress influenza neuraminidase completely to reach, and then suppress copying of influenza virus.
In another detailed description of the invention of the present invention, the pathological symptom that the icariin of preparation and derivatives for treatment thereof or flu-prevention virus cause or disease.Described influenza virus comprises by influenza virus A type, B-mode and the third type and their various hypotypes, serotype, genotype, gene mutation precursor virus; One or more the disease that the influenza syndrome that described pathological symptom or disease are caused by the various hypotype of above-mentioned virus forms.Be preferably cold syndrome, cough, fever, sneeze, nasal obstruction, stream clear water sample nasal mucus, fear of cold, headache, weak, systemic pain, pneumonia, tonsillitis, pharyngitis, laryngitis, shed tears, hoarseness, breathing syndrome, cardiopulmonary disease, cold limbs, nausea and vomiting, dry cough, dry mouth and tougue, dizzy sleepy, diarrhoea, flu miscarriage, myocarditis, hypertension, myocardial infarction, apoplexy, bronchia mucosal hyperemia, edema, there are secretions, pharyngeal mild hyperaemia, submandibular lymph nodes enlargement, pharyngeal congestion edema.
Term used herein " treatment " or refer to treatment treatment measures and prevention or the measure that prevents, wherein, prevent or slow down the pathological symptom suffered from or the imbalance of (weakening) experimenter.If the icariin received according to the therapeutic dose of method as herein described and derivant thereof, then the symptom of experimenter succeed " treatment ", experimenter shows, and one or more signs and the symptom that can observe and/or be measured to symptom reduce and disappear.It is also understood that treatment described herein or prevent the various patterns of medical conditions to be intended to represent " significantly ", it comprises all treatments or prevention and is less than all treatments or prevention, wherein reaches and is correlated with certain biology or result that medical science is correlated with.
In various embodiments, carry out suitable external test or in vivoassay and measure whether be suitable for treatment based on the effect of the medicine of specific icariin and derivant thereof and its administration.In various embodiments, external test can be carried out to the representative cell involved by the disease of experimenter, measure whether the given medicine based on icariin and derivant thereof creates expectation effect to described Virus Type.If the icariin received according to the therapeutic dose of method as herein described and derivant thereof, then the symptom of experimenter is treated with succeeding, i.e. experimenter's display, one or more signs and the symptom that can observe and/or be measured to above-mentioned pathological symptom or disease reduce and disappear.It is also understood that treatment described herein or prevent the various patterns of medical conditions to be intended to represent " significantly ", it comprises all treatments or prevention and is less than all treatments or prevention, wherein reaches and is correlated with certain biology or result that medical science is correlated with.
In a specific embodiment of the present invention, experimenter is used to the medicine containing icariin and derivant thereof preventing and/or treating effective dose.
Before testing human experimenter, can test Candidate compounds used in treatment in suitable animal model system, described animal subjects model system is including, but not limited to non-human primates (such as baboon, orangutan, monkey); The such as pet animals of cat, Canis familiaris L., Serpentis etc.; The farm-animals of pig, horse, cattle, goat etc.; Such as any animal of the laboratory animal of rat, mice, monkey, rabbit etc.In one embodiment, icariin and derivant thereof are administered to the experimenter of (state of this symptom that is easy to get) in the danger suffering from or be in above-mentioned pathological symptom or disease, attempt to improve the one or more factors causing this pathological symptom or disease.
Term used herein " effective dose " refer to be enough to obtain needed for treat and/or prevent effect, such as cause prevention or alleviate the amount with the symptom of pathological symptom or disease.The amount being administered to the compositions of experimenter by the character of the type and seriousness and individuality that depend on disease, such as health condition at ordinary times, age, sex, body weight and the tolerance to medicine.Described amount also depends on extent, seriousness and type.Those skilled in the art can according to these factors and other be because usually determining suitable dosage.Described compositions also can carry out administration in conjunction with one or more other therapeutic compound.In method described herein, compound of the present invention can be administered to has one or more signs of pathological symptom or the experimenter of pathological symptom.Such as, " treatment effective dose " refers to the average level of the physiological action of minimally mitigation symptoms.
Usually, described dosage can prevent or alleviate the seriousness for the treatment of situation or indication or expansion.Correct dosage will depend on environment, and whether such as treated situation, administration time table, described compound are used separately or be combined with another kind of therapeutic agent is used, the plasma half-life of described compound and the holistic health of experimenter.
Can adopt comprise oral, external, suck, per nasal, per rectum, percutaneous or injection method of application to experimenter's medication.
In a specific embodiment of the present invention, prepare and contained for described compound disclosed herein, daily dosage is that about 0.1mg/kg elects 0.1mg/kg extremely about 500mg/kg as to about 10000mg/kg body weight, preferably about 0.1mg/kg to about 1000mg/kg body weight, more, most preferably is 0.5mg/kg extremely about 200mg/kg body weight.Every day can use 1 to 6 time, and preferred every day uses 2 or 3 times.Interval also can be irregular, those skilled in the art generally acknowledge that the optimised quantity of compound or its pharmaceutically acceptable salt and the interval of single-dose are by the nature and extent by situation to be treated, the form of administration, approach and site and the concrete condition of experimenter for the treatment of and determine, and most preferably scheme can be determined by routine techniques.Those skilled in the art be also to be understood that the optimum process for the treatment of, namely in given natural law every day the compound that gives or the administration quantity of its pharmaceutically acceptable salt can utilize conventional treatment method of testing by those skilled in the art and determine.In treatment use, in relatively short interval, relatively high dosage is had to be needs sometimes, until the process of disease slows down or stops, and preferably until subject shows partial or fully improve described disease or pathological symptom.Therefore, patient can to the administering mode of effecting prevention property of patient.Those skilled in the art will recognize that, some factor can affect the dosage and time of effectively treating an experimenter, including, but not limited to disease or the order of severity of imbalance, previous treatment, health condition and/or the age of experimenter and the Other diseases of existence.And, utilize the treatment reference composition for the treatment of effective dose described herein to treat an experimenter and can comprise single therapy or a series for the treatment of.
Compound of the present invention can be used together with other compounds of at least one.Described compound can be used as independent preparation or be combined in unit dosage forms simultaneously, or uses in turn.Under any circumstance, multiple therapeutic agent can with any order or administration even simultaneously.If side by side, described multiple therapeutic agent can be provided as single, unified form or with multiple form (such as, as several different preparation of single tablet or separately independently tablet or capsule, drop pill).A kind of therapeutic agent in described therapeutic agent may provide with multiple dose type, or wherein several can provide as multiple dose type.If the interval not side by side, then between multidose can from be greater than 0 week to be less than about 1 week, be less than about 2 weeks, be less than about 4 weeks, be less than about 2 months, be less than about 4 months or be even less than about half a year scope change.
Term used herein " unit dosage forms " refers to the unit be separated physically of the unit dose be suitable as humans and animals experimenter, each unit comprise independent scheduled volume compound or with other agent combination, its amount is enough to produce desired effects with pharmaceutically acceptable diluent, carrier or media mix as calculated.
In the specific embodiment of the present invention, preparation is containing the pharmaceutical composition of icariin or derivatives thereof, and it is used for the treatment of and/or prevents the flu disease caused by influenza virus to improve relevant pathological symptom or disease.It, by the mode such as oral, injection, inhalation, plays a role to snibject.Therefore pharmaceutical composition of the present invention can be prepared into various dosage form, such as, the compositions for oral medication can be capsule, drop pill, gel, tablet, powder, granule, buccal tablet, effervescent tablet, syrup, emulsion, the prepared product of controlled release, rapidly-soluble prepared product, oral liquid dosage forms etc.; Compositions for external medication can be liniment type medicine, cream, external application, ointment, lotion, the gentle spray of liquid spray etc.Compositions for inhalation administration can be solution, dispersion liquid, dry powder etc.; Also can extract highly purified icariin compound from Chinese herbal medicine Herba Epimedii, the nanometer icariin of preparation liposome, is prepared into ejection preparation and extended drug form.In described pharmaceutical composition, icariin or its derivant proportion, account for the gross weight of whole preparation for 0.0%-99.0%, preferably 0.0%-90% with weight, be more preferably 10%-80%, most preferably be 25-70%.
In pharmaceutical preparation of the present invention, for oral formulations, pharmaceutically acceptable adjuvant used including, but not limited to lubricant and cosolvent, as magnesium stearate, calcium stearate, zinc stearate, stearic acid, Glyceryl Behenate, sodium stearyl fumarate, Talcum, silica gel; Disintegrating agent, as starch, cyclodextrin, carboxymethyl cellulose, cross-linked carboxymethyl cellulose, crospolyvinylpyrrolidone, gelatin; Diluent or compression agent, as lactose, sucrose, mannitol xylitol, erythritol, Sorbitol, microcrystalline Cellulose; Flavouring agent or other composition, as Fructus Fragariae Ananssae, Fructus Citri tangerinae, Fructus Musae, Herba Menthae, Mel; Cosolvent, as drinks (medicated beer, fruit wine, rice wine, Chinese liquor), vegetable oil, animal oil, ethanol, isopropyl alcohol, glycerol, propylene glycol, benzyl alcohol, dimethyl sub-wind, Polyethylene Glycol, arabic gum, lecithin, pyrrolidone, oleic acid, azone.
Carrier granular can be crystal or the spheroid of lactose or sucrose; Or composite sphere or granule, as by making sucrose granulate the spheroid of the sugar formed with starch as binding agent, the calcium carbonate spheroid formed as binding agent by starch or maltodextrin.Carrier granular also can be the granule of any other medical acceptable excipient, as hydroxy propyl cellulose crude granule, guar gum granule, xanthan gum granule.Carrier can have various ways, and all these adjustment of selecting and measuring are obviously in the limit of power of those skilled in the art.
By peroral route, can comprise according to the compound of icariin or derivatives thereof of the present invention or its compositions and be dissolved in food or health product liquid, as being dissolved in liquid or the water-ol class solution of the aqueous of seasoning alternatively.It can be included into the solid excipient that can swallow, and such as, in particle form, pill, tablet, in the form of enteric coatel tablets.It also can be placed in the liquid in food or health product, and itself optional condition is in swallowable capsule.For swallowing, can be multiple Orally administered composition embodiment, particularly, can be the accrete Orally administered composition embodiment of food.Enteric coatel tablets are manufactured, colloid capsule, gel, emulsion, tablet, capsule or solution and preparation by the method for routine.Particularly, other form any of dietary supplement ingredient or condensed food or health product can be included according to activating agent of the present invention, such as food or health product rod, or compression or incompressible powder among.These powder can dilute with water, can at soda, and dilute with water in salt cheese production or soybean derivatives, maybe can bring in food or health product rod.
Recorded by following embodiment, understand the present invention better, but be not limited to this embodiment.
Embodiment
The preparation of embodiment 1. influenza neuraminidase and icariin thereof suppress influenza neuraminidase activity analysis
Get influenza virus H 5 N 1 (A/chicken/Guangdong/174/04 (H5N1)), the H9N2 (A/chicken/Guangdong/SS/94 (H9N2) of 104TCID50, in order to attempt the inhibition of icariin whether to other hypotype neuraminidase, also have selected H9N2 Strain and prepare neuraminidase liquid), add triton x-100 (Sigma) to 1% concentration, prepare influenza neuraminidase liquid, measure for inhibitor activity.The icariin standard substance (ChromaDex company of the U.S.) of purity 98%, the sub-wind (Sigma) of the dimethyl with 30% is dissolved, and concentration is 10mmol/L;
Methylumbelliferyl-a-D-N-acetylneuramimic acid (MUNANA, sigma) is made into 20 micromoles/L concentration; Oseltamivir (Oseltamivir, ChromaDex company of the U.S.) is configured to the concentration of 10mmol/L with water; The buffer (325mmol MES (Sigma), 40mmol CaCl (Sigma), pH6.5) of 10 times.Enzyme assay reaction system is 100 microlitres, carries out in black 96 orifice plate.Influenza neuraminidase liquid adds the icariin of variable concentrations in advance, add 3 μ L reaction buffers again, water 27 μ L, is wherein provided with the icariin Concentraton gradient (end reaction system concentration) of 1 μm of ol, 5 μm of ol, 50 μm of ol, 100 μm of ol, 200 μm of ol.Establish Oseltamivir to compare experiment as stated above, end reaction system concentration gradient is 1 μm of ol, 5 μm of ol, 50 μm of ol, 100 μm of ol, 200 μm of ol simultaneously.In addition, if (other composition adds by above-mentioned concentration not add influenza neuraminidase liquid, influenza neuraminidase liquid water consumption substitution), (other composition adds by above-mentioned concentration not add icariin and Oseltamivir, with water postreaction volume), do not add icariin and Oseltamivir but add 1% DMSO (other composition adds by above-mentioned concentration, the DMSO of 1% is end reaction system concentration) control experiment, adding MUNANA each reaction system front is 40 μ L, and each reaction system refers to table 1.37 DEG C of reactions are after 1 hour, and all reaction systems add 60 microlitre 20 micromoles/L MUNANA, and 37 DEG C are reacted 1 hour, stop responding with the sodium hydroxide of 0.002mol/L.With the exciting light of super microplate reader (Synergy HT, blo-tek, USA) 360 and 460 utilizing emitted light under measure fluorescence intensity.
Table 1, add the one-tenth of each reaction system before MUNANA and be grouped into
Influenza neuraminidase activity analysis (embodiment 1) is suppressed by icariin, disclosing icariin suppresses influenza neuraminidase activity very strong, than high nearly 40 times of Oseltamivir, prompting icariin can, as the candidate lead compound preventing and/or treating flu virus disease, be expected to develop the better medicine of a kind of Oseltamivir.In addition, this experiment also demonstrate that icariin can suppress the neuraminidase activity of different subtype too well, namely can suppress N1 (H5N1) and N2 (H9N2) enzymatic activity, prompting icariin may suppress the neuraminidase activity of various hypotype.
Icariin infected by influenza inhibitory action in embodiment 2. cell in vitro level
Select MDCK (Madin-Darby Canine Kidney) cell (Harbin Veterinary Medicine Inst., China Academy of Agriculture provides) as proliferation of influenza virus cell, DMEM (Gibco company) is as mdck cell basal medium.According to the methods such as Xie (Yi Xie, James A.Schafer.Inhibition of ENaC by intracellular Cl
-in an MDCK clone with high ENaC expression.Am J Physiol Renal Physiol.2004Oct; 287 (4): F722-31.), mdck cell cultivation is carried out: DMEM culture fluid 190mL adds hyclone 20mL, blue or green chain dual anti-2mL, 7.5%NaHCO
3regulate pH to be 7.4, above-mentioned each component mixing is MDCK complete culture solution, get 15 generation cell bottle, add Digestive system appropriate, be advisable just to cover cell, and put down about 10min in 37 DEG C of incubators gently, treat the contracting of cell circle, when not mutually being connected, inhaling gently and abandon Digestive system, add culture fluid, repeatedly gently inhale piping and druming, take off wall to cell, become even, counting cells, 1.0 × 10
5/ mL cell is inoculated in new 25mL culture bottle, puts 37 DEG C, 5%CO
2environment quiescent culture.Influenza virus is inoculated in 9 ~ 11 age in days chick embryo allantoic cavities, hatch 48 ~ 72h for 35 DEG C, then Embryo Gallus domesticus is put 4 DEG C of refrigerator overnight, next day gathers in the crops allantoic fluid, the centrifugal 10min of 3000rpm, discards precipitation, gets supernatant and carries out blood coagulation tests, determine that its titre is greater than 1:64 person and gathers in the crops supernatant ,-80 DEG C of preservations.Influenza infection cell determination virus titer: cultured cell goes down to posterity and counts, and is transferred in 6 well culture plates, adjusting every porocyte density is 1.0 × 10
5/ mL.Get H5N1 hypotype, H9N2 (in order to attempt the inhibition of icariin whether to other hypotype neuraminidase, we have selected H9N2 Strain simultaneously) subtype influenza virus 1mL joins wherein in A and B porocyte respectively, and C hole adds normal saline and makes negative control.Every hole cultivating system final volume is adjusted to 5mL, A and B porocyte hatches 72h totally 3 groups, often organizes and repeats 5 times again, and basis of microscopic observation cytopathy (Cytopathic effect, CPE) situation, determines virus titer.With reference to method (the Shuji Hatakeyama of Hatakeyama etc., Yuko Sakai-Tagawa, Maki Kiso, Hideo Goto, Chiharu Kawakami, Keiko Mitamura, Norio Sugaya, Yasuo Suzuki and Yoshihiro Kawaoka.Enhanced Expression of an α 2, 6-Linked Sialic Acid on MDCK Cells Improves Isolation of Human Influenza Viruses and Evaluation of Their Sensitivity to a Neuraminidase Inhibitor.J Clin Microbiol.Aug 2005, 43 (8): 4139 – 4146.), carry out icariin for influenza virus Inhibition test: icariin dissolves with the sub-wind (sigma) of dimethyl of 30%, concentration is 1mmol/mL, join in DMEM cell culture medium respectively, the experimental result of reference example 1, carry out detection icariin suppression virus replication experiment detection if concentration is 5 μm of ol, substitute with PBS the negative control that icariin makes to suppress virus replication.Use 96 well culture plates, if 5 parallel laboratory tests, 1.0 × 10
5/ mL mdck cell 100 μ L, what add 100 μ L contains 200TCID
50the virus of titre and the icariin of various concentration, put 37 DEG C, 5%CO2 environment quiescent culture 3 days, observe viral increment situation.Use flow cytometer (Flow cytometry respectively, Beckman Coulter Inc. of the U.S.), with reference to method (the Liu H of Liu etc., Zhang GL, Shen L, Zeng Z, Zhou BL, Liu CH, Nie G.Application and evaluation of a pseudotyped virus assay for screening herbs for anti-H5N1avian influenza virus.Zhong Xi Yi Jie He Xue Bao.2010Nov, 8 (11): 1036-40.), fluorescent antibody (Anti-Influenza A H5N1Hemagglutinin Antibody and Anti-Influenza A H9N2Hemagglutinin Antibody, purchased from Sino Biotlogical Inc.), with reference to method (the Ebrahimi SM of Ebrahimi etc., Tebianian M, Aghaiypour K, Nili H, Mirjalili A.Prokaryotic expression and characterization of avian influenza A virus M2gene as a candidate for universal recombinant vaccine against influenza A subtypes, specially H5N1and H9N2.Mol Biol Rep.2010Jul, 37 (6): 2909-14.), utilize staining to analyze virus multiplication situation, fluorescence microscope (German Olympus) detects.Real-time n cell functional analysis instrument (xCELLigence RTCA MP, ACEA Biosciences Inc) analyze: 200TCID50/mL influenza virus H 5 N 1 and H9N2 are mixed according to 1:1 ratio with the inhibitor diluted, be placed in 37 DEG C of incubators to carry out hatching 1 hour, then virus is added to the mixture of inhibitor and has grown in 96 porocyte detection of electrons plates of mdck cell, be placed in 37 DEG C of incubators and cultivate 190-200 hour.During experiment, be provided with 7 icariin Concentraton gradient, namely culture medium contains 1 μm of ol, 5 μm of ol, 10 μm of ol, 20 μm of ol, 50 μm of ol, 100 μm of ol, 200 μm of ol, and is provided with viral negative control and not containing the contrast of virus inoculation of icariin.Use real-time n cell function to divide instrument to carry out cultivating and detecting, every 15min carries out a data acquisition.
Icariin infected by influenza inhibitory action research (embodiment 2) in cell in vitro level; experiment is destroyed from influenza virus by the pathogenicity of mdck cell and icariin protection mdck cell; comprise and use fluorescent antibody to detect the sick infection cell of infection influenza, flow cytometry analysis and n cell functional analysis instrument analysis in real time; demonstrate icariin all well and can suppress proliferation of influenza virus, protect cell from viral subversive.In this experiment, we used the Strain of two kinds of influenza neuraminidases, i.e. H5N1 (N1) and H9N2 (N2), the Strain of icariin to these two kinds different neuraminidases shows almost equal inhibition, show that the Strain of neuraminidase may be resisiting influenza virus hypotype, serotype, the genotypic inhibitor compound of a wide spectrum, icariin can as medicine, food, the health product preventing and/or treating pathological symptom that various types of influenza virus causes or disease.Embodiment 3. icariin suppresses the zoopery of influenza virus
3.1 laboratory animal kind and features
BALB/C mice, Harbin Veterinary Medicine Inst., China Academy of Agriculture's Experimental Animal Center breeding, Quality of Experimental Animals credit number is Heilungkiang-SYXK 2007-167.
9 weeks ~ 10 weeks age, about body weight 20g, male and female half and half.
3.2 rearing conditions
Animal Lab. air timing ventilation, illumination well, keep implementing room room temperature.5 animals raised by every cage, feed with specially for the expanded pellet diet that mice makes, freely drink water.Before experiment starts, observe animal feed, active situation and feces thereof etc. one week, the mouse of a unsoundness is chosen as next step experiment.
The animal safety evaluation of 3.3 icariin
First-selection, uses acute toxicity test to carry out safety evaluatio to icariin, to provide foundation for the application of icariin.This experiment adopts acute toxicity test maximum tolerated dose method, first chooses the healthy BALB/C mice 20 of about body weight 20g, male and female half and half.The route of administration of clinical plan is oral, and capital stock experiment have employed administration by gavage administration.Icariin distilled water is modulated into grume, and 1 time gavage gives mice (about every Mus 200 milligrams of icariin), and before gavage, animal overnight fasting, freely drinks water.Give normal diet after gavage, observe poisoning symptom, death condition, weigh weekly once, the observation period is 2 weeks.Under similarity condition, get non-icariin interpolation normal diet feeding and observe as a control group with batch mouse.Experimental result result shows all mices and is any poisoning symptom of appearance, proves that icariin belongs to innocuous substance.
Icariin is to influenza virus infection zooprophylazis Experiment on therapy: select male BALB/c mouse in 6 week age, often organize mice and inoculate the H5N1 virus of 50 μ L 102LD50 respectively (because H9N2 is for the pathogenic instability of mice, so this experiment does not select H9N2) (experimental group 1 and experimental group 2), negative control (experimental group 4) is made with PBS, and establish the matched group (experimental group 3) giving separately icariin, the mouse infection of virus inoculation is after 4 hours, if 4 laboratory animal groups, often organize 10 mices, every Mouse oral administration 1mg, be equivalent to oral icariin 25 mg/kg body weight every day, every day 2 times sooner or later, totally 14 days, weigh Mouse Weight every day 1 time, observe Mouse Weight change and survival rate situation.Histopathologic examination: select to die of illness, treatment group, lung tissue 10% neutral formalin (Neutral-buffered formalin) that per os gives merely icariin and normal mouse is fixed, then revise and use the ethanol dehydration of variable concentrations, dimethylbenzene (Xylene, Sima) after soaking, with paraffin (Paraffin, Sigma) sealing is inlayed, be cut into the thin slice of 3 micron thickness, be placed on microscope slide, dewax continuously with ethanol, with hematoxylin (Hematoxylin) and Yihong (Eosin, Sigma) dye, observe under optical microscope (Light microscopy).
Table 2, zoopery grouping experiment situation
| Group | Virus inoculation strain | Per os gives icariin | PBS |
| Experimental group 1 | Infect | No | No |
| Experimental group 2 | Infect | Be | No |
| Experimental group 3 | Do not infect | Be | No |
| Experimental group 4 | Do not infect | No | Be |
Icariin suppresses the zoopery (embodiment 3) of influenza virus, and result display icariin can protect the infection of high pathogenic avian influenza virus (H5N1).In addition, tissue pathological slice shows, and icariin can also promote lung tissue immune cell infiltrate, improves the immune protection effect of lung tissue.Icariin may be used for the icariin prepared and derivatives for treatment thereof or prevents medicine, food, the health product of the pathological symptom that caused by influenza infection or disease.
The invention has the beneficial effects as follows: icariin can suppress the activity of influenza neuraminidase, thus suppress the copying of influenza virus, can be used for influenza infection and cause various prevention and treatment of diseases medicines, food, health product exploitation.
The present invention does not limit to the particular implementation described in this application, as the unitary declaration of independent aspect of the present invention.Various amendment and change is carried out when it will be understood by those skilled in the art that the spirit and scope that can not depart from the application.According to above description, except enumerating herein, the functionally equivalent purposes in the scope of the present disclosure is obvious for those skilled in the art of this area.Such change and amendment are intended to fall within the scope of the appended claims.The disclosure only by claims and with such claim the four corner that is equal to of scope limit.Should be appreciated that the disclosure is not limited to specific method, reagent, compositions and biosystem, certainly, described method, reagent, compositions and biosystem can change.It can also be appreciated that term used herein only for describing specific embodiment, it is restrictive for not being used for.
In addition, when describe in the mode of Ma Kushi group feature of the present disclosure or in, this area one of skill in the art will appreciate that the disclosure is also with described by the mode of the arbitrary single member in Ma Kushi group or subgroup member.
All patents that are referred herein or that quote, patent application, earlier application and publication are incorporated to herein by reference and in full, comprise all accompanying drawings and form, they are not contradicted with the clearly instruction of this description.Other embodiment is proposed in right.
Claims (10)
1. icariin and derivant thereof are for the preparation of the purposes prevented and/or treated in the medicine of human or animal's pathological symptom that influenza virus causes or disease, food, health product or cosmetics, it is characterized in that described icariin and derivant thereof be respectively following formula (I) and (II) define, comprise its independent isomer, the raceme of isomer or non-racemic mixture, oxide or its pharmaceutically acceptable salt and hydrate
Wherein,
Xa, Xb, Xc, Xd, Xe, Xf, Xg, Xh, Xi, X
1, X
2, X
3, X
4, X
5, X
6and X
7independent is oxygen atom (O) or sulphur atom (S);
Alkyl (the C that Ra, Rb, Rc, Rd, Re, Rf, Rg, Rh and Ri are independently hydrogen atom (H), 1-5Ry substitutes
1-C
10) or (C
1-C
10)-O-(C
1-C
10), each alkyl (C
1-C
10) replace with 1-5 R y, each R y is Rq or (C
2-C
10) alkyl, (C
3-C
10) alkyl, (C
3-C
10) cycloalkyl-alkyl, (C
8-C
14) bicyclic alkyl alkyl, (C
8-C
14) tricyclic alkyl alkyl ,-(C
5-C
10) cycloalkyl-alkyl, (C
3-C
10) cycloalkyl, (C
8-C
14) bicyclic alkyl alkyl, (C
8-C
14) tricyclic alkyl alkyl, phenyl, naphthyl, (C
14) aromatic group, they each can replace with one or more Rz, and each Rz is (C
1-C
6) alkyl, (C
2– C
6) thiazolinyl, (C
1– C
6) alkynyl, (C
3– C
8) cycloalkyl, (C
6-C
8) cycloalkenyl group, phenyl, 3-to 5-unit) heterocycle, C (halogen)
3or CH (halogen)
2;
R
1, R
2, R
2, R
4, R
5, R
6, R
7and R
8independent is hydrogen atom (H), or (C
1-C
10) alkyl, (C
2-C
10) thiazolinyl, (C
2-C
10) alkynyl, (C
3-C
10) cycloalkyl, (C
8-C
14) bicyclic alkyl alkyl, (C
8-C
14) tricyclic alkyl alkyl, phenyl, naphthyl, (C
14) aromatic group, they each can be that non-substituted, one or more Rz or Rq replaces, the one or more Rq replacement replaced with more than 1 Rz, and each Rq can be CN, OH, halogen, N
3, NO
2, N (Rz)
2, ═ NR z, CH ═ NRz, NRz OH, ORz, CORz, C (O) ORz, OC (O) Rz, OC (O) ORz, SRz, S (O) Rz or S (O)
2rz;
Y
1, Y
2and Y
3independent is CH
3, (C
1-C
12) alkyl, also can with Ry combination replace (C
1-C
12) alkyl, each (C
1-C
12) alkyl can replace with 1-5 R y or an equilibrium ion replaces.
2. purposes as claimed in claim 1, it is characterized in that described pathological symptom or disease comprise by pass cough, fever, sneeze, nasal obstruction, stream clear water sample nasal mucus, fear of cold, headache, weak, systemic pain, pneumonia, tonsillitis, pharyngitis, laryngitis, shed tears, hoarseness, breathing syndrome, cardiopulmonary disease, cold limbs, nausea and vomiting, dry cough, dry mouth and tougue, dizzy sleepy, diarrhoea, flu miscarriage, myocarditis, hypertension, myocardial infarction, apoplexy, bronchia mucosal is congested, edema, there is secretions, pharyngeal mild hyperaemia, submandibular lymph nodes enlargement, pharyngeal congestion edema, one or more disease of the group of common cold syndrome composition.
3. purposes as claimed in claim 1 or 2, to is characterized in that described icariin and derivant thereof by oral, external, suction, per nasal, per rectum, percutaneous or injecting pathway snibject.
4. purposes as claimed in claim 3, is characterized in that described experimenter comprises the animal of people, farm-animals, pet animals, laboratory animal.
5. purposes as claimed in claim 4, is characterized in that described experimenter has suffered from or suffered from or has been in the pathological symptom of influenza infection or the risk of disease.
6. purposes as claimed in claim 5, is characterized in that described purposes also comprises and uses to described experimenter the medicine containing icariin and derivant thereof preventing and/or treating effective dose.
7. purposes as claimed in claim 6, is characterized in that described purposes also comprises the pharmaceutical composition containing the icariin mixed with pharmaceutically accepted diluent or carrier and derivant thereof.
8. purposes as claimed in claim 7, is characterized in that described pharmaceutical composition to be prepared into the dosage form being suitable for oral, external, injection, described experimenter is used to the drug combination preparation of the effective dose prevented and/or treated.
9. purposes as claimed in claim 1 or 2, it is characterized in that described purposes also comprises and the compositions containing icariin and derivant thereof is prepared into the form being suitable for food, health product or cosmetics, cause the experimenter described in the pathological symptom of human or animal or the risk of disease to use the described compositions of the effective dose prevented and/or treated to suffering from or suffering from or being in influenza infection.
10. the purposes as described in claim 1-9, is characterized in that described purposes also comprises and is used for the compositions containing icariin and derivant thereof preventing and/or treating the various pathological symptom or disease that the various hypotype of influenza virus, serotype, genotype and their gene mutation body thereof cause.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1799593A (en) * | 2004-12-31 | 2006-07-12 | 中国医学科学院药用植物研究所 | Medicine for preventing and treating bird flu and its preparation process |
| CN1969870A (en) * | 2005-11-24 | 2007-05-30 | 北京奇源益德药物研究所 | Antivirus compound formulation, preparation process, quality control method and use thereof |
| CN103083670A (en) * | 2012-10-26 | 2013-05-08 | 通化新海生物产业有限公司 | Application of inhibiting agent of cathepsin K |
| CN103265591A (en) * | 2013-05-24 | 2013-08-28 | 四川大学 | Icariin derivative, and preparation method and application thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1732925A (en) * | 2005-08-02 | 2006-02-15 | 天津药物研究院 | Sustained release minipills of lovastatin and its preparation process |
| CN103621567A (en) * | 2013-11-19 | 2014-03-12 | 青岛茂丰有机蔬菜有限公司 | Drug for preventing and treating gummosis |
-
2015
- 2015-04-19 CN CN201510184295.7A patent/CN104857007A/en active Pending
- 2015-12-15 WO PCT/CN2015/097322 patent/WO2016110173A1/en active Application Filing
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1799593A (en) * | 2004-12-31 | 2006-07-12 | 中国医学科学院药用植物研究所 | Medicine for preventing and treating bird flu and its preparation process |
| CN1969870A (en) * | 2005-11-24 | 2007-05-30 | 北京奇源益德药物研究所 | Antivirus compound formulation, preparation process, quality control method and use thereof |
| CN103083670A (en) * | 2012-10-26 | 2013-05-08 | 通化新海生物产业有限公司 | Application of inhibiting agent of cathepsin K |
| CN103265591A (en) * | 2013-05-24 | 2013-08-28 | 四川大学 | Icariin derivative, and preparation method and application thereof |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105727269A (en) * | 2016-03-22 | 2016-07-06 | 中国药科大学 | Application of neuraminidase and inhibitors thereof to myocardial ischemia and myocardial infarction |
| WO2017161594A1 (en) * | 2016-03-22 | 2017-09-28 | 齐炼文 | Applications of neuraminidase and inhibitor thereof in myocardial ischemia and myocardial infarction |
| CN105727269B (en) * | 2016-03-22 | 2018-10-02 | 中国药科大学 | The application of neuraminidase and its inhibitor in myocardial ischemia and myocardial infarction |
| US10639289B2 (en) | 2016-03-22 | 2020-05-05 | China Pharmaceutical University | Application of neuraminidase and inhibitors thereof in myocardial ischemia and myocardial infarction |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2016110173A1 (en) | 2016-07-14 |
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