CN101622265A - Antiviral compound - Google Patents
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- CN101622265A CN101622265A CN200880006407A CN200880006407A CN101622265A CN 101622265 A CN101622265 A CN 101622265A CN 200880006407 A CN200880006407 A CN 200880006407A CN 200880006407 A CN200880006407 A CN 200880006407A CN 101622265 A CN101622265 A CN 101622265A
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Abstract
The invention provides neuraminidase (NA), erythrocyte agglutination element (HA), have structural protein M
2Virus demonstrate inhibiting novel cpd.These compounds also can be used as the inhibitor of hepatitis C virus (HVC) replicator.The present invention has also described and suitable pharmaceutically acceptable carrier blended compound of the present invention alone or in combination.
Description
Technical field
The present invention relates to as neuraminidase (NA), erythrocyte agglutination element (HA), structural protein M
2Merge the novel cpd of the inhibitor of (HEF) glycoprotein with the hemagglutinin esterase; Comprise the pharmaceutical composition that is used for the treatment of, prevents or improve the described compound of virus infection; And use described compound and method for compositions.
Have been found that influenza virus demonstrates diameter when observing be 80 to 120nm spheroid form and/or filament shape (Yoshinori Fujiyoshi et al. " Finestructure if influenza A virus observed by electroncryo-microscopy " under electron microscope, The EMBO Journal 13 (2), p 318-26,1994).The characteristic feature of viromembrane is, has radial ridge (radialprojections) [(Wilson I.A.et al. " Structure of thehaemoagglutinin membrane glycoprotein of influenza virus under the situation corresponding to the A type of erythrocyte agglutination element (HA) and neuraminidase (NA) and Type B virus
Resolution ", Nature, 289, p 366-73,1981; Varghese J.N.et al. " Structure of the influenza virus glycoprotein antigenneuraminidase at
Resolution ", Nature, 303, p 35-40,1983; Colman P.M.et al. " Structure of the catalytic and antigenicsites in influenza virus neuraminidase ", Nature, 303, p41-44,1983)], and under situation, produce three kinds of biologic activity: be attached to acceptor (H) corresponding to the C type influenza virus that is called hemagglutinin esterase fusion (HEF) glycoprotein, acceptor inactivation (E) and fusion (F) (Herrier Georg, et al., " A synthetic sialic acid analogueis recognized by influenza Cvirus as a receptor but is resistantto the receptor-destroying enzyme ", J.Biol.Chem., 2567 (8), p 12501-12505,1992).
First-generation anti-virus product (mainly being adamantane derivative, similar amantadine, Rimantadine etc.) it is believed that the M by blocking-up A type influenza virus
2-protein ion passage and working.Blocking-up H
+Ion passes through M
2The inflow of-protein ion passage can suppress to shell and the release of free ribonucleoprotein in tenuigenin.This only finds in the A C-type virus C, but does not take place in Type B virus.
After this, antiviral strategy relates to for example exploitation of zanamivir and oseltamivir of s-generation antiviral agent, and their suppress erythrocyte agglutination element (HA) or neuraminidase (NA), and it is present in the surface of A type and Type B influenza virus with the mushroom ridge.Sialic acid part and the glycolipid class film surface that be attached to the target cell that will infect of these albumen by the cracking sialoglycoprotein.In addition, at the terminal point of virus replication, it is essential that neuraminidase discharges virus for cracking sialic acid from acceptor.
By contrast, the strategy of current inhibition hepatitis C virus (HVC) is included in and uses ribavirin (as monophosphate) to suppress by level in the reduction cell in synthesizing of single phosphoric acid guanosine-.In addition, ribavirin (for triphosphate) is by reducing virus of A RNm and synthesizing of ARN polysaccharase suppressing ARNm-guanidine radicals transferring enzyme (guanilyltranspherase).
Much more more and more international application has reported the resistance strains of influenza viruses, and lasting sudden change (particularly A C-type virus C) and drug resistant variants or its combination are transmissible and are morbific fully.In this respect, in recent years, scientists is described as the material risk that the world pop disease is broken out with A type bird flu (H5N1) virus.
Similarly, hepatitis C infection is very general in the whole world, therefore the serious pathogenic situation of also having represented a kind of patient's of influence number to increase.
Therefore, to be badly in need of developing the antiviral compound of improvement, preferably there is the Multiple Combination mechanism to the virus replication effect in it.
Background technology
Itzstein, M.von et al.; " Nature ", 363 (6428), the appropriate design based on the influenza virus replication inhibitors of sialidase is disclosed among the p 418-423 (1993).Colman, P.M.et al.; WO 92/06691 (PCT/AU90/00501, open day: on April 30th, 1992), Itzstein, L.M.von et al.; EP 0539204A1 (European application 92309684.6, open day: on April 28th, 1993), and Itzstein, L.M.von et al.; WO 91/16320 (PCT/AU91/00161, open day: disclose compound on October 31st, 1991), and it is believed that and demonstrate the interior resisting virus activity in conjunction with neuraminidase.Bischofberger N.W.et al.; US 5,952,375 (U. S. application 08/606,624, the applyings date: on February 26th, 1996) disclose the new compound as neuraminidase inhibitor.
Babu Y.S., Chad P., Bantia S.et al.: " Discovery of a novel; highly potent; orally active; and selective influenzaneuraminidase inhibitor through structure-based drug design " .J Med Chem, 43 (19): 3482 (2000), International Application No. WO 99/33781 (international application no PCT/US 98/26871, open day: on July 8th, 1999) disclose new substituted compound and the derivative that is used as neuraminidase inhibitor.
In addition, infect in order to treat chronic HVC, for example be described among Martindale 33.rd Ed. (2002) the p 639-43, common medical practice is combined administration purine nucleoside analogs ribavirin or Viramidine (trade(brand)name) and cytokine Interferon Alpha-2b (or glycol interferon alpha-2b) for example for example simultaneously in people experimenter.In fact, it is believed that single phosphoric acid ribavirin and this analog derivative suppress to concentrate in synthesizing of single phosphoric acid bird (purine) nucleosides and the cell, and triphosphate RNA interfering m-guanidine radicals transferring enzyme.
Goal of the invention
An object of the present invention is to provide virus is particularly shown effective inhibiting new compound to influenza virus and hepatitis virus.This compounds is to membranin, erythrocyte agglutination element (HA), the structural protein M in the virus for example
2, and glycolytic enzyme for example neuraminidase (NA) produce its combination and restraining effect optionally, more particularly work by disturbing with viral neuraminidase.These new compounds also produce hepatitis C virus (HVC) and suppress active.The most commonly encountered diseases poison transmittance process inhibitor that another purpose provides improvement and cheaper and causes serious virus infection, and do not have any cross resistance with the antiviral agent of present use.Further purpose provides the method for the reasonable combination of using new compound of the present invention or they and other known antiviral agent again.Another purpose provides the pharmaceutical composition that is used for above purpose.
The whole consideration according to the present invention to those skilled in the art, it is obvious that the purpose of these and other will become.
Summary of the invention
In a first aspect of the present invention, this paper provides general formula (I) compound:
Wherein:
X is-CH
2-,-O-,-CHF-,-CF
2-;
The C of singly-bound or two key shacks
2-C
3
R
1Expression-OH, halogen or-the B/ part, condition is the C that has two key shacks
2-C
3The time R
1Do not exist; And
R
2Expression-OH ,-O-CH (C
2H
5)
2,-NH
2,-NHC (NH) NH
2Or-NH-OH; And
R
3Be-NH
2,-NHCO-CH
3Or-NH-CO-CH
2-OH; And
R
4Expression-CHOH-CHOH-CH
2-OH ,-CHOH-CH
2-B/ part ,-CH
2-B/ part or-the B/ part
Wherein-B/ partly represents:
-B1/ part-B2/ part
Wherein:
R
5Expression connection functional group-NH-,-CH
2-NH-,-CH (CH
3)-NH-,-NH-CH (CH
3)-NH-,-C (CH
3)
2-CH
2-NH-,-NH-CO-CH
2-O-CH
2-CH
2-NH-or
R
6Be-H ,-CH
3Or-C
2H
5And
R
7Be-H ,-CH
3Or-C
2H
5And
R
8Expression connect functional group-NH-,-CO-NH-or-C (NH)-NH-;
And their C1-4 carboxyl list or polyester, additive salt, solvate, the enantiomorph of fractionation and the diastereomer of purifying.
The present invention also comprises pharmaceutical composition, and it contains independent The compounds of this invention or makes up with other promoting agent in being applicable to the Mammals pharmaceutically acceptable carrier that particularly people uses.
In another embodiment of the present invention, neuraminidase and/or albumen M
2The activity method that can be may further comprise the steps suppress: suspect with The compounds of this invention or compositions-treated and contain neuraminidase and/or albumen M
2Sample.
The present invention provides the method that is used for the virus infection that for example caused by influenza virus or hepatitis virus in host treatment or prevention on the other hand, and this method comprises by any suitable route of administration gives the The compounds of this invention disclosed herein of host's administering therapeutic effective dose.
In other embodiment of the present invention, also provide the novel method of synthetic The compounds of this invention.
Embodiment
The present invention relates to structural formula (I) compound of following configuration:
Wherein:
X is-CH
2-,-O-,-CHF-,-CF
2-;
The C of singly-bound or two key shacks
2-C
3And
R
1Expression-OH, halogen or-the B/ part, condition is the C that has two key shacks
2-C
3The time R
1Do not exist; And
R
2Be-OH ,-O-CH (C
2H
5)
2,-NH
2,-NHC (NH) NH
2Or-NH-OH; And
R
3Be-NH
2,-NHCO-CH
3Or-NH-CO-CH
2-OH; And
R
4Expression-CHOH-CHOH-CH
2-OH ,-CHOH-CH
2-B/ part ,-CH
2-B/ part or-the B/ part
Wherein-B/ partly represents:
-B1/ part-B2/ part
Wherein:
R
5Expression connection functional group-NH-,-CH
2-NH-,-CH (CH
3)-NH-,-NH-CH (CH
3)-NH-,-C (CH
3)
2-CH
2-NH-,-NH-CO-CH
2-O-CH
2-CH
2-NH-or
R
6Be-H ,-CH
3Or-C
2H
5And
R
7Be-H ,-CH
3Or-C
2H
5And
R
8Expression connect functional group-NH-,-CO-NH-or-C (NH)-NH-.
In preferred embodiments, X represents that by-O-it is typical sialic acid ring, wherein at C
1On carboxylic acid keep replacing because it is considered to the sorption site of viral neuraminidase.In another preferred embodiment, the C of singly-bound or two key shacks
2-C
3, condition is, when having two key, and R
1Do not exist.In another typical embodiment, R at least
1Or R
4Can by-the B/ part is mono-substituted.Therefore, R
1Preferred expression-OH, halogen or typical-B/ part are worked as R
4Expression-CHOH-CHOH-CH
2-OH or typical-CHOH-CH
2During-B/ part, wherein should-B/ part can be-the B1/ part or-the B2/ part.
Typical when using-during the B1/ part, A and B strains of influenza viruses suppress mechanism to the specificity of neuraminidase preferably to be brought out, and M
2The blocking-up of-protein ion passage (existing only in the A strain) is also allowed.-B2/ part also can be used for monosubstituted derivative, to realize similar restraining effect.In another suitable combination, R
1And R
4Can be identical or different dibasic.In preferred combination, R
1And R
4Can be all partly replaced by-B2-, the gained compound it is believed that more optionally suppressing HVC duplicates.
Work as R
1Quilt-B1/ part or-B2/ partly replaces and R
4Different surface is shown-the B2/ part or-during the B1/ part, it is believed that the gained restraining effect can influence A and B strains of influenza viruses, and can influence HVC.
In further preferably making up, work as R
1Or R
4Be connected to simultaneously main structure (I)-when B1/ partly replaces, R
2Can be-OH and R
3Can be-NHCO-CH
3Another preferred embodiment is to connect the R of functional group
5Can be-NH-or-CH (CH
3)-NH-, and R
6And R
7Can be preferably-H or-CH
3Work as R
1Or R
4Main structure (I)-when B2/ partly represented, the connection R8 of functional group can be preferably-CO-NH-by being connected to.
The present invention also comprises the straight or branched C of The compounds of this invention
1-4The enantiomorph of carboxyl monoesters or polyester, additive salt, solvate, fractionation and the diastereomer of purifying.
In another embodiment of the present invention, neuraminidase and/or M
2The activity of ionic channel can contain neuraminidase and/or M by suspecting with The compounds of this invention or compositions-treated
2Proteic viral sample suppresses or blocks.
Another aspect of the present invention relates in mammalian hosts the particularly method of influenza A and B virus strain or HVC of treatment or prophylaxis of viral infections, and it comprises the The compounds of this invention of giving host's administering therapeutic effective dose by any suitable route of administration.
The novel method of synthetic The compounds of this invention also is provided in another embodiment.
The present invention also comprises pharmaceutical composition, and it contains independent The compounds of this invention for example being applicable to give in the pharmaceutically acceptable carrier of administration, perhaps with for example combination of another compound of the present invention or one or more promoting agents.
The further aspect of the present invention comprises treatment or prevents the method for described virus disease or illness, this method is by mammiferous treatment that is used in combination The compounds of this invention or pharmaceutical composition or its mixture and while or is used alternatingly the treatment that another treats the promoting agent of effective dose that described promoting agent also can suppress this type of virus infection.
The present invention also comprises the method that produces The compounds of this invention.Described compound can be by any suitable technology preparation in the organic synthesis, and they are well known to a person skilled in the art.
For R
1Be directly connected to the C of sialic acid ring
2The generality preparation of preferred compound of the present invention (X=-O-), preferred reductive amidation methodology, as by Tang et al.Biochem.Biophys.Res.Commun., the early stage method of describing of 132:474-80 (1985), and by Stoll et al., Biochem.J., 256:661-4 (1988) and by ScharzmannG.et al., Biochem., the method for 22:5041-9 (1983) modification.In fact, by according to above method, can produce the C of sialic acid ring
2Be connected with the covalency amine between the amino group of-B/ part.
Similarly, for other preferred compound of the present invention (R for example
4Be directly connected to the C of sialic acid ring
6(X=-O-) general preparation method those compounds) can be followed many known technologies of describing in the following publication: Compendium of Organic SyntheticMethods (John Wiley ﹠amp; Sons, New York), Vol.1, Ian T; Harrisonand Shuye, Harrison, 1971; Vol.2, Ian T.Harrison and ShuyenHarrison, 1974; Vol.3, Louis S.Hegedus and Leroy Wade, 1977; Vol.4, Leroy G.Wade, jr., 1980; Vol.5, Leroy G.Wade, Jr.1984; And Vol.6, Michael B.Smith; And March, J., 2AdvancedOrganic Chemistry, the 3rd edition, (John Wiley ﹠amp; Sons, New York, 1985), Comprehensive Organic Synthesis.Selectivity, Strategy ﹠amp; Efficacy in modern Organic Chemistry.In 9 Volumes, Barry M.Trost, Editor-in-Chief (Pergamon Press, New York, printing in 1993).
The exemplary method of many preparation The compounds of this invention is provided below, but they will be not can the restricted application method scope.
Usually, for example temperature, reaction times, solvent, treatment process etc. are common those in this area for the concrete reaction of carrying out to reaction conditions.Comprise the detailed description of above-mentioned condition in document material of being quoted and the material of wherein quoting.As exemplary method, below general step can be used for R of the present invention
4The generality of the compound that replaces is synthetic.
Yet, it will be understood to those of skill in the art that other standard operation can be used for producing identical material.
The preparation of the intermediate 1 of the following structure of step 1-:
Under refluxing, in methyl alcohol, use Dowex 50 (H
+) the stir process sialic acid reaches 24-48 hour time.Leach resin, filtrate is concentrated into drying, (intermediate 1) that the dimethyl that obtains needing replaces.
The preparation of the intermediate 2 of the following structure of step 2-:
The solid that obtains from step 1 restores with aqueous sodium hydroxide solution, and at room temperature stirs, and continues 1-3 hour usually.Then with this mixture Dowex 50 (H
+) resin moderated to pH7.0-7.5.Filter, then make the filtrate lyophilize, obtain corresponding non-methylated ester (intermediate 2).
The intermediate 3 of the following structure of step 3-and 4 preparation:
The intermediate that obtains from step 2 was in the dark reacted 1 hour with different mol ratios with the sodium periodate aqueous solution, produce intermediate 3 (lower ratio) or intermediate 4 (higher ratio).Then barium acetate is added in the throw out, again by removing by filter excessive iodate and periodate.Then with the filtrate lyophilize.Obtain little yellow solid (intermediate 3 or 4).
The intermediate 5 of the following structure of step 4-and 6 preparation:
Intermediate 5
Intermediate 6
Under agitation will be dissolved in respectively from the intermediate 3 or 4 of step 3 gained in the aqueous formic acid, heat 1 hour down at 80 ℃ again.With the lyophilize separately of gained solution.Obtain intermediate 6 and 5 respectively.
Step 5-prepares final R according to following exemplary reaction scheme
4Mono-substituted compound
Option A
The reaction product of intermediate 5 generation-B1/ part
R of the present invention
4Mono-substituted (B1/ part) compound
Option b
The reaction product of intermediate 6 generation-B2/ part
Compound 2
Another R of the present invention
4Mono-substituted (B2/ part) compound
Will from each intermediate that obtains 5 or 6 of step 4 use respectively distilled water and selected generation required-reaction product of B/ part (B1/ part or-B2/ part) restores, and makes reaction mixture keep standing over night down at 4 ℃.Add sodium borohydride then, and make reaction mixture at room temperature keep leaving standstill to reach 1 hour.Make each sample at Dowex 50 (H
+) deionization on the post, again with the elutriant lyophilize, obtain the final compound of different the present invention respectively.
Other typical R of the present invention
4Mono-substituted (having different-B/ part) compound for example is expressed as follows, and describes in a further embodiment:
Compound 3
Compound 4
Compound 5
Compound 6
Compound 7
Compound 8
Compound 9
Compound 10
Another preferably prepares embodiment and comprises by using following reactions steps to synthesize R
1Mono-substituted compound:
Step 1-R
1The preparation of mono-substituted (B/ part) compound
Sialic acid is dissolved in the mixture of first alcohol and water of different ratios (preferred 9: 1), under gentle agitation, adds generations-B/ reaction product partly again.Add after the sodium borohydride, this mixture was kept 2 hours under 60 ℃ and gentle agitation.Make this mixture by Dowex50 (H then
+) post, so that sodium borohydride changes into boric acid.With the elutriant lyophilize,, pass through paper filter again with the methyl alcohol recovery of aliquot.Isolate resistates, make the elutriant drying.Repeat the operation several (usually more than 5 times) of back as required.The gained solid finally is dissolved in the aliquot water, and lyophilize then obtains the R of the present invention that needs
1Mono-substituted (having-the B/ part) compound.
Other typical reaction scheme can be expressed as follows:
Scheme C
The reaction product of sialic acid (intermediate) generation-B1/ part
R of the present invention
1Mono-substituted (B1/ part) compound (compound 11)
Scheme D
The reaction product of sialic acid generation-B2/ part
Another R of the present invention
1Mono-substituted (B2/ part) compound
Another other preferably prepare embodiment and comprise by adopting following general reaction scheme to synthesize R
1And R
4Dibasic compound:
Step 1-R
1And R
4The preparation of dibasic (B/ part) compound
By in conjunction with life side described above scheme (A, B, C and D), those skilled in the art can obtain disubstituted compound of the present invention.
The present invention's typically finally disubstituted (having identical or different-B/ part) compound is expressed as follows:
Typical R
1And R
4Disubstituted (having identical-B1/ part) compound
Another typical R
1And R
4Disubstituted (having identical-B2/ part) compound
It will be obvious to those skilled in the art that by use identical generation-B1/ part or-reaction product of B2/ part, it provides the R as general formula (I) expression separately
6, R
7And R
8Other possible combination, this may obtain similar bisubstituted compound of the present invention.
Typical R of the present invention
1And R
4Disubstituted (have various combination-B/ part) compound
Another typical R of the present invention
1And R
4Disubstituted (have various combination-B/ part) compound
Similarly, by use combination-B1/ part and-the differential responses product of B2/ part, it provides the R that represents as general formula (I) separately
6, R
7And R
8Other possible combination, this may obtain the different bisubstituted compound of similar the present invention.
Be apparent that to those skilled in the art, by using classical Resorcinol-HC l method (Svennerholm L. " Quantitative estimation of sialic acids.II.A colorimetric resorcinol-hydrochloric acid method. " Biochem.Biophys.Acta, 24 (3),: 604-611,1957; Miettinen J.etal. " Use of butyl acetate in determination of sialic acid. ", Acta Chem.Scand., 13,856-858,1959) and TBA (2-thiobarbituricacid) method (Warren L. " The thiobarbituric acid assay of sialic acids. ", J.Biol.Chem., 234,1971-5,1959; Aminoff D. " Methods for thequantitative estimation of N-acetylneuraminic acid and theirapplication to hydrolysates of sialomucoids. ", Biochem.J., 81 (2), 384-392,1961) detect and the quantitative assay sialic acid because of following former thereby different: Resorcinol-HCl method for example all can detect and quantitative assay during sialic acid sugar compounds (sialoglycocompounds) conjugation in sialic acid and the free existence of derivative thereof and with other sugar.By contrast, the TBA method only is being connected to 2 (C
2) (R of hydroxyl-partly of carbon atom
1=-OH) is not substituted Shi Caike and detects and quantitative assay sialic acid and derivative thereof.
The compounds of this invention also is included in optical isomer enrichment or that split at any or all of asymmetric atom place.Racemic mixture and non-enantiomer mixture, and separation or synthetic, essentially no other enantiomorph to or the right single optical isomer of diastereomer, all within the scope of the present invention.Can by known technology with racemic mixture be separated into they single, be essentially optically pure isomer, described technology for example, make with the optical activity auxiliary for example the diastereoisomeric salt that forms of acid or alkali separate, then transform and be returned to optically active substance.In most of the cases, the optical isomer of Xu Yaoing is to begin to react synthetic with stereospecificity by the suitable steric isomer from required initial substance.
The optional acceptable non-toxic salt of salt, especially pharmacy that comprises this paper compound of the present composition, it comprises for example inorganic or preferred organic acid or alkali.When needing the water-soluble salt of compound, salification is preferred methodology.
The present invention relates on the other hand and suppresses the active method of neuraminidase, and this method comprises that handling suspection with The compounds of this invention contains for example step of the sample of viral neuraminidase of neuraminidase.It is believed that The compounds of this invention can be used as the inhibitor of neuraminidase,, perhaps have other purposes described below as the intermediate of this type of inhibitor.In fact, described inhibitor can be attached in the surface or inner chamber of neuraminidase, and these surfaces or inner chamber have uniqueness how much for neuraminidase.Yet, in conjunction with the compound of neuraminidase can be in various degree reversible combination.These basically the compound of irreversible fixation be the ideal candidates person who is used for the inventive method.The organism that contains neuraminidase comprises bacterium (vibrio cholerae, clostridium perfringens, streptococcus pneumoniae and Arthrobacter sialophilus) and virus (especially orthomyxovirus or Paramyxo virus for example influenza virus A and B, parainfluenza virus, rhinovirus, coronavirus, coronavirus mutant and/or improvement coronavirus, mumps virus, Avian pneumo-encephalitis virus, ewcastle disease virus and Sendai virus).From these organic that obtain any or active restraining effect of neuraminidase of finding within the scope of the invention.The compounds of this invention also is used among the animal or human treatment or prevents above-mentioned infection, and described animal is duck, rodent or pig for example.
In further embodiment, The compounds of this invention is to come the inhibition activity of its neuraminidase is screened by the routine techniques of estimating enzymic activity.In the context of the invention, at first screen exemplary compounds at the vitro inhibition neuraminidase.
The further aspect of the present invention relates to blocking-up H
+Ion is discharged into cytoplasmic method by inflow, inhibition shelling and the free ribonucleoprotein of M2-protein ion passage, and this method comprises that handling suspection with The compounds of this invention contains for example step of the sample of influenza virus A of M2-albumen.In fact, think that also The compounds of this invention is the onset by blocking virus M2-protein function.
Another further aspect of the present invention relates to the synthetic method that suppresses single phosphoric acid guanosine-and RNAm-guanidine radicals transferring enzyme, and this method comprises by handling with The compounds of this invention is suspected that sample is synthetic with RNAm and the RNA polymerase of blocking-up HVC.
In another embodiment, it is believed that of the inhibitor onset of different disubstituted compound while of the present invention as neuraminidase and M2-protein ion passage.
The compounds of this invention can be prepared with conventional carrier and vehicle, and these carriers and vehicle will be selected according to conventional practice.Tablet will contain vehicle, glidant, weighting agent, tackiness agent etc.Aqueous solution preparation is made into sterile preparation, and in the time need coming administration by the mode except that oral, they are normally isoosmotic.All preparations will be chosen wantonly and contain vehicle, for example at known publication " Handbook of Pharmaceutical Excipients ", and the 4th edition, those that describe among the RoweR.C.et al., Pharmaceutical Press (2003).Vehicle comprises xitix and other oxidation inhibitor, and sequestrant is for example dextrin, hydroxy alkyl cellulose, hydroxyalkyl methylcellulose gum, stearic acid etc. of EDTA, carbohydrate for example.
One or more compounds of the present invention (the following activeconstituents that also is called) can be used by any approach that is suitable for illness to be treated.Suitable approach comprises mouth, rectum, nose, part (comprising oral cavity and hypogloeeis), vagina and parenteral (comprise subcutaneous, intramuscular, vein, intracutaneous, sheath is interior and epidural) etc.Be appreciated that preferred approach can change with for example experimenter's illness.The advantage of The compounds of this invention is that they are that oral biology is effective, and can be with the oral Pharmaceutical dosage forms administration; Can but must not give them by approach in the lung or in the nose.
Though it is possible that activeconstituents of the present invention is used separately, they preferably exist with pharmaceutical preparation.Preparation of the present invention, it can for animals or human, comprises the activeconstituents of the present invention or the compound of at least a above-mentioned definition, and one or more acceptable carriers and other optional therapeutic component.Described carrier must be " acceptable ", represents that other composition of itself and preparation is compatible and to being nontoxic on its experimenter's physiology.
Preparation comprises those that are applicable to aforementioned route of administration.Preparation can exist with unit dosage form expediently, and can prepare by the known any method of pharmaceutical field.
Preparation technique is found in " Remington ' s Pharmaceutical Sciences " Mack Publishing Co., Easton, Pa., U.S.A. usually.These class methods comprise makes activeconstituents and carrier-bound step, this carrier by a kind of or what time ancillary component constitute.Usually, said preparation can evenly and densely the two combines with the solid carrier of liquid vehicle or dispersion and fining or this by making activeconstituents, and then, if desired, it is desired that product is configured as.
Be applicable to that oral preparation of the present invention can be prepared as solid unit for example capsule, cachet or tablet, it contains the activeconstituents of predetermined amount separately; Powder or particle; Solution in waterborne liquid or non-aqueous liquid or suspension; Oil-in-water liquid emulsion or water-in-oil liquid emulsion.Activeconstituents can also exist with bolus, Electuary or paste.
Tablet is suppressed or molded preparations with one or more ancillary components by optional.Compressed tablets can for example powder or particle prepare optional and tackiness agent, lubricant, inert thinner, sanitas, tensio-active agent or dispersant by activeconstituents being pressed into free-pouring form on suitable machine.Molded tablet can be by will preparing with the mixture pressing mold of the powdered activeconstituents of inert liquid diluent humidifying on suitable machine.Tablet can be chosen wantonly by dressing or indentation, and optional being mixed with provides activeconstituents from wherein slowly discharging or sustained release.
For eye or other outside organization for example mouthful and the infection of skin, said preparation can be preferably used as local with ointment or emulsifiable paste, the amount that described ointment or emulsifiable paste contain activeconstituents is for for example 0.075 to 20%w/w (comprising that scope is 0.1% to 20% activeconstituents, increment is 0.1%w/w, for example 0.6%w/w, 0.7%w/w etc.), be preferably 0.2 to 15%w/w, most preferably be 0.5 to 10%w/w.When being mixed with ointment, activeconstituents can use with paraffin matrix or water dispersible ointment base.Perhaps, this activeconstituents can be mixed with emulsifiable paste with the oil-in-water emulsifiable paste matrix.
If desired, the water of emulsifiable paste matrix can comprise, for example, the polyvalent alcohol of 30%w/w at least, the alcohol that promptly has two or more hydroxyls is propylene glycol, 1,3 butylene glycol, N.F,USP MANNITOL, sorbyl alcohol, glycerine and polyoxyethylene glycol (comprising PEG 400) and composition thereof for example.Topical formulations can comprise desirably that also the promotion activeconstituents is by skin or other compound of being attacked the zone absorption or permeating.The example of this type of dermal osmosis accelerator comprises methyl-sulphoxide and analogue.The oil phase of emulsion of the present invention can be made of in a known way principal component.Though this phase can only comprise emulsifying agent (being referred to as emulgent in addition), its expectation comprise at least a emulsifying agent and fat or oil or with fat and the two mixture of oil.Hydrophilic emulsifier preferably uses with the lipophilic emulsifier as stablizer.Also preferably include oil ﹠ fat the two.Simultaneously, have or the emulsifying agent that do not have a stablizer is prepared into so-called emulsifying wax, and this wax is prepared into so-called emulsification ointment base with oil ﹠ fat, this emulsification ointment base forms the oily disperse phase of cream formulation.
The emulgent and the emulsion stablizer that are applicable to preparation of the present invention comprise
60,
80, cetostearyl alcohol, phenylcarbinol, tetradecyl alcohol, Zerol and Sulfuric acid,monododecyl ester, sodium salt.
Be used for the suitable oil of said preparation or fatty selection according to realizing that the cosmetic properties of expecting carries out.This emulsifiable paste should be preferably product non-oily, non-contamination and that can wash and have suitable denseness with avoid from the pipe or other container leak.Can use straight or branched, list or dialkyl for example two-dissident, two acid esters, stearic acid isocetyl ester, coconut fatty acid propylene glycol diesters, isopropyl myristate, decyl oleate, Wickenol 111, butyl stearate, palmitinic acid 2-(ethyl hexyl) ester or be called the mixture of side chain/ester of Crodamol CAP, last three kinds is preferred ester.They can use separately or use according to required properties of combination.Perhaps, use for example white vaseline of high-melting-point lipid.
Preparation for topical application to eye also comprises eye drops, and wherein dissolved the or appropriate carrier that is suspended in activeconstituents of activeconstituents is especially in the aqueous solvent.Activeconstituents preferably is present in this type of preparation with 0.5 to 20% concentration, advantageously is 0.5 to 10%, particularly about 2.0%w/w.
Preparation for topical application to mouth comprises, lozenge, and it contains the activeconstituents that is in the flavoured base (being generally sucrose, gum arabic or tragacanth gum); Pastille, it contains the activeconstituents that is in the inert matrix (for example gelatin and glycerine, perhaps sucrose and gum arabic); And mouth wash shua, it contains the activeconstituents that is in the appropriate liquid carrier.
The preparation that is used for rectal administration can exist with the suppository with suitable matrix, and this matrix comprises for example cocoa butter or salicylate.
Be used in the lung or granularity that the preparation of intranasal administration has 0.1 to 500 micrometer range (comprises that scope is 0.1 to 500 micron a granularity, the increment micron for example is 0.5,1,30 micron, 35 microns etc.), thus it is used by sucking fast through nasal meatus or sucking the arrival alveolar sac by per os.Suitable preparation comprises the water-based or the oily solution of activeconstituents.Be applicable to that the preparation that aerosol or dry powder doses are used can prepare according to conventional methods, and can be with other therapeutical agent administration, up to the present this other therapeutical agent for example is used for the compound that treatment as described below or flu-prevention A or B are infected.
The preparation that is used for vaginal application can exist with vaginal suppository, tampon agent, ointment, gelifying agent, paste, foaming agent or sprays, and these preparations also contain suitable carrier known in the art except activeconstituents.
Be applicable to that the preparation that parenteral is used can comprise water-based and non-aqueous aseptic injectable solution, it can contain oxidation inhibitor, buffer reagent, fungistat and provide and expect experimenter's the solute of the isoosmotic preparation of blood; And the water-based and the non-aqueous sterile suspension that can comprise suspension agent and thickening material.
Preparation can be present in single dose or the multi-dose container, for example in sealed ampoule bottle or the phial, and can preserve under lyophilize (freeze-drying) condition, only need face with the preceding for example water for injection of sterile liquid carrier (solvent) that adds immediately.Facing injection allocation, to penetrate solution and suspension be sterilized powder, particle or tablet preparation from aforesaid kind.Preferred unit dose formulations be contain activeconstituents as this paper dosage every day recited above or unit sub-doses every day, the perhaps formulation of its suitable umber.
Should be appreciated that preparation of the present invention can comprise other reagent that the preparation type with being discussed of this area routine is relevant except the top composition of mentioning especially, for example be suitable for those Orally administered reagent, comprise seasonings.
The present invention further provides veterinary composition, it comprises at least a activeconstituents as defined above and the animal doctor uses carrier.
The animal doctor is the material that can be used for using the said composition purpose with carrier, can be solid, liquid or gaseous matter, its be inert or veterinary applications acceptable, and compatible with activeconstituents.These veterinary compositions can be oral, parenteral is used, and perhaps uses by the approach of any other expectation.
The compounds of this invention can be used for providing controlled-release pharmaceutical formulation, it contains one or more The compounds of this invention (" controlled release preparation ") as activeconstituents, the release of activeconstituents is controlled and regulates in said preparation, so that administration frequency is lower or improved the pharmacokinetics or the toxicity of given activeconstituents.
The effective dose of activeconstituents depends on sanatory character, toxicity at least, and no matter whether this compound is used for prevention (than low dosage) no matter or suppress active influenza infection, also medication and pharmaceutical preparation, and the effective dose of activeconstituents will be used routine dose to increase progressively ratio research by the clinician to come definite.
Expectation is that every day about 0.0001 is to about 100mg/kg body weight.Typically be every day about 0.01 to about 10mg/kg body weight.More typically be that every day about 0.01 is to about 5mg/kg body weight.More typically be that every day about 0.05 is to about 0.5mg/kg body weight.For example for inhalation, the per day for adults recommended dose of about 70kg body weight will be 1mg to 1000mg, be preferably 5mg to＞500
Mg, and can be the form of single dose or multiple doses.When experimenter's pathologic condition needs, also can use the effective per daily dose of bigger treatment.
Activeconstituents of the present invention (or compound) also can be used in combination with other activeconstituents.The illness of being treated of selecting this type of combination to be based on, the cross reactivity of each composition and the pharmaceutical properties of combination.For example, when the virus infection of treatment respiratory system particularly during influenza infection, composition of the present invention and antiviral agent (for example amantadine, Rimantadine and ribavirin), mucolytic, expectorant, bronchodilator, microbiotic, febrifuge or anodyne make up.Usually, microbiotic, febrifuge and anodyne are used with The compounds of this invention.
The detailed description of the present invention is enough to make that those skilled in the art can prepare and use the theme material of following examples.Obviously, some modification of the method and composition of following examples can be carried out in the scope of the invention and spirit.The present invention will be further described with reference to following exemplary embodiment and appended Fig. 1-4.
Description of drawings
Fig. 1 is FT-I R (Fourier transform infrared spectroscopy) spectrum (A) of compound 1, initial in contrast intermediate amantadine (B),
Fig. 2 is the FT-IR spectrum (A) of compound 4, initial in contrast intermediate Rimantadine (B),
Fig. 3 is the FT-IR spectrum (A) of compound 8, initial in contrast intermediate memantine (B),
Fig. 4 is the FT-IR spectrum (A) of compound 10, initial in contrast intermediate ribavirin (B).
Embodiment
The preparation of intermediate 1
The sialic acid (0.73mmol) that is dissolved in the 225mg in the absolute methanol of 40ml is mixed into the Dowex 50 (H of 0.5g+) in the resin. Continuing to make this mixture reflux 48 under the stirring Hour. By resorcinol HCl and thiobarbituricacidα-(TBA) assay determination as can be known, 24 During with 48 hours, there is respectively 85% and 97% sialic acid to change into intermediate 1. Then will Resin filter and is concentrated into drying with eluent by rotary evaporator to general paper filter, Obtain the little yellow liquid of oily. Then with the ether of this oily liquids with a small amount of volume: methyl alcohol (3: Mixture 1w/w) restores. Under 4 ℃, make this solution keep leaving standstill 24-48 hour, by Filter and collect the crystalline deposit thing, use again P2O
5Dry. Obtain the solid intermediate 1 (M.W. of 145mg 337.4) (yield: 60.0%).
1 pair of resorcinol of gained intermediate-HCl reaction aobvious positive (identical with sialic acid intensity), And aobvious negative to the TBA reaction.
Embodiment 2
The preparation of intermediate 2
Make the intermediate 1 (0.43mmol) of 145mg be dissolved in the 0.06M hydrogen-oxygen of 10.7ml Change in the sodium water solution, at room temperature continue again to stir 2-3 hour. Then this mixture is used Dowex 50 (H+) resin moderated to pH7.0-7.5. Leach resin, eluent is freezing dried Dry, the solid of the look that obtains turning white. Obtain the solid intermediate 2 (M.W.323.3) of 132.6mg (yield: 95.0%).
2 pairs of resorcinols of gained intermediate-HCl reaction aobvious positive (identical with sialic acid intensity), And aobvious negative to the TBA reaction.
Embodiment 3
The preparation of intermediate 3
The solid (intermediate 2) of the freeze-drying of 132.6mg (0.41mmo l) is dissolved in 4.3ml Distilled water in. The 0.038M sodium metaperiodate aqueous solution (NaIO that adds then every part of 10.7ml4) (0.41mmol) (mol ratio=1: 1) at room temperature in the dark holds this solution again The continuous stirring 1 hour. The 0.1M barium acetate aqueous solution of 12.8ml is added in this mixture, So that excessive iodate and periodate precipitation. Use general paper filter that this mixture is filtered. It is saturated that this eluent is advertised into carbon dioxide, so that excessive barium acetate precipitation, then paper using Filter filters. The eluent freeze drying obtains little yellow solid. Collect the solid of 101.9mg Intermediate 3 (M.W.291.3) (yield: 85.0%).
3 pairs of resorcinols of gained intermediate-HCl reaction aobvious positive (identical with sialic acid intensity), And aobvious negative to the TBA reaction.
Embodiment 4
The preparation of intermediate 4
Make the solid (intermediate 2) of the freeze-drying of 132.6mg (0.41mmol) be dissolved in 4.3ml Distilled water in. 0.2M sodium metaperiodate (NaIO with 10.7ml4) aqueous solution (2.14mmol) (mol ratio: 1: 5.24) adds in this solution, makes it at room temperature in the dark continue to stir 1 Hour.
The 0.1M barium acetate aqueous solution of 12.8ml is added in this mixture, so that excessive Iodate and periodate precipitation. Use general paper filter that this mixture is filtered. Make eluent Advertise into carbon dioxide saturatedly, to be settled out excessive barium acetate, filter at paper filter then. The eluent freeze drying obtains little yellow solid. Collect the solid intermediate 4 (M.W. of 91.4mg 261.23) (yield: 85.0%).
4 pairs of resorcinols of gained intermediate-HCl reaction aobvious positive (identical with sialic acid intensity), And aobvious negative to the TBA reaction.
Embodiment 5
The preparation of intermediate 5
Make the powder intermediate 4 of the freeze-drying of 91.43mg (0.35mmol) be dissolved in pH about 4.0 The 2.3mM aqueous formic acid of 2.0ml in. Make solution be heated to 80 ℃ and reach 1 hour. So After make the solution freeze drying.
Obtain intermediate 5 (the M.W.247.20) (yield: 84.1%) of 72.76mg.
5 couples of resorcinol-HCl of gained intermediate and TBA reaction are all aobvious positive.
Embodiment 6
The preparation of intermediate 6
Make the solid intermediate 3 of the freeze-drying of 101.96mg (0.35mmol) be dissolved in pH about 4.0 The 2.3mM aqueous formic acid of 2.0ml in. Make solution be heated to 80 ℃ and reach 1 hour. So After make the solution freeze drying.
Obtain intermediate 6 (the M.W.277.23) (yield: 96.7%) of 93.83mg.
6 couples of resorcinol-HCl of gained intermediate and TBA reaction are all aobvious positive.
Embodiment 7
The preparation of compound 1
Make the solid (intermediate 5) of the freeze-drying of 84.0mg (0.34mmol) be dissolved in 5.0ml Distilled water in. The amantadine that under slowly stirring, adds then 51.442mg (0.34mmol). slowly stir and 4 ℃ under this mixture stirring is spent the night. Add 40.0mg Behind the sodium borohydride, reactant mixture was kept 1 hour. Make then this mixing Thing is by Dowex 50 (H+) resin column, make excessive sodium borohydride change into boric acid. Make wash-out The liquid freeze drying. With aliquot methyl alcohol cryodesiccated solid is restored then, make the eluent drying. This operation is repeated 3 times at least. Dry solid is finally restored with aliquot water, then freezing Drying obtains final compound 1.
Obtain compound 1 (the M.W.382.45) (yield: 85.0%) of 110.53mg.
1 pair of resorcinol of compound of collecting-HCl reaction and TBA reaction are all aobvious positive. Chemical combination The FT-IR of thing 1 (FFIR) spectrum (A) is plotted in Fig. 1, with it (B) of contrast Be initial intermediate amantadine, show following reference value: a) acid amides I bands of a spectrum: 1640cm-1 B) acid amides II bands of a spectrum: 1550cm-1 C) primary amino radical group: 600-800cm-1，1590-650cm
-1，
3330-3380cm
-1 Parahelium group: 700-800cm-1；1615cm
-1；3300cm
-1。
Embodiment 8
The preparation of compound 2
Make the solid (intermediate 6) of the freeze-drying of 94.26mg (0.34mmol) be dissolved in 5.0ml Distilled water in. The Ribavirin (0.34mmol) that under slowly stirring, adds then 82.7mg. This mixture is spent the night 4 ℃ of lower slowly stirrings. After adding the sodium borohydride of 40.0mg, make Reactant mixture kept 1 hour under stirring and room temperature. Make then this mixture pass through Dowex 50 (H+) resin column, so that excessive sodium borohydride changes into boric acid. Make the eluent freeze drying. With aliquot methyl alcohol the freeze drying thing is restored then, make eluent again dry. This operates repetition At least 3 times. Dry solid finally is dissolved in the aliquot water, and then freeze drying obtains Solid (compound 2). The solid chemical compound 2 (M.W.505.4) of collection 94.02mg (yield: 89.0%).
The 2 couples of resorcinol-HCl of compound that collect and TBA reaction are all aobvious positive.
Embodiment 9
The preparation of compound 3
Make the solid (intermediate 6) of the freeze-drying of 94.26mg (0.34mmol) be dissolved in 5.0ml Distilled water in. The amantadine that under slowly stirring, adds then 51.442mg (0.34mmol). This mixture is spent the night 4 ℃ of lower slowly stirrings. Add the boron of 40.0mg Behind the sodium hydride, reactant mixture was at room temperature stirred 1 hour. This mixture is passed through Dowex 50 (H+) post, in order to make excessive sodium borohydride change into boric acid. Make eluent freezing Dry. Then with cryodesiccated dissolution of solid in aliquot methyl alcohol, make the filtrate drying. This behaviour Repeat at least 3 times. Then with the dissolution of solid of drying in aliquot water, then freeze drying, Obtain final compound 3. Collecting the solid chemical compound 3 (M.W.412.5) of 122.02mg (receives Rate: 87.0%).
The 3 couples of resorcinol-HCl of compound that collect and TBA reaction are all aobvious positive.
Embodiment 10
The preparation of compound 4
Make the solid (intermediate 5) of the freeze-drying of 84.0mg (0.34mmol) be dissolved in 5.0ml Distilled water in. The Rimantadine that under slowly stirring, adds then 60.962mg (0.34mmol). This mixture is spent the night 4 ℃ of lower slowly stirrings. Add the boron of 40.0mg Behind the sodium hydride, reactant mixture was at room temperature stirred 1 hour. This mixture is passed through Dowex 50 (H+) post, so that excessive sodium borohydride changes into boric acid. Make eluent freezing dried Dry. With aliquot methyl alcohol cryodesiccated solid is restored then, make the eluent drying. This operation Repeat at least 3 times. With the powder water-soluble solution of aliquot of drying, then freeze drying gets then To final compound 4.
Collect compound 4 (the M.W.410.5) (yield: 90.0%) of 125.61mg.
4 pairs of resorcinols of compound of collecting-HCl reaction and TBA reaction are all aobvious positive. Chemical combination The FT-IR spectrum (A) of thing 4 is plotted in Fig. 2, and (B) of contrast is initial intermediate Buddha's warrior attendant second with it Amine shows following reference value: a) acid amides I bands of a spectrum: 1640cm-1 B) acid amides II bands of a spectrum: 1550cm-1 C) primary amino radical group: 600-800cm-1，1590-650cm
-1，3330-3380cm
-1 Parahelium group: 700-800cm-1；1615cm
-1；3300cm
-1。
Embodiment 11
The preparation of compound 5
Make the solid (intermediate 6) of the freeze-drying of 94.26mg (0.34mmol) be dissolved in 5.0ml Distilled water in. The Rimantadine that under slowly stirring, adds then 60.962mg (0.34mmol). This mixture is spent the night 4 ℃ of lower slowly stirrings. Add the boron of 40.0mg Behind the sodium hydride, reactant mixture was at room temperature stirred 1 hour. This mixture is passed through Dowex 50 (H+) post, in order to make excessive sodium borohydride change into boric acid. Make eluent freezing Dry. The freeze drying thing is dissolved in the aliquot methyl alcohol, makes the eluent drying. This operates repetition At least 3 times. Make then the dry solid water-soluble solution of aliquot, then freeze drying obtains Whole compound 5. Collecting the final solid chemical compound 5 (M.W.440.5) of 137.79mg (receives Rate: 92.0%).
The 5 couples of resorcinol-HCl of compound that collect and TBA reaction are all aobvious positive.
Embodiment 12
The preparation of compound 6
The solid (intermediate 5) of the freeze-drying of 84.0mg (0.34mmol) is dissolved in 5.0ml Distilled water in. The somantadine (0.34mmol) that under slowly stirring, adds then 70.5mg. This mixture is spent the night 4 ℃ of lower slowly stirrings. After adding the sodium borohydride of 40.0mg, make Reactant mixture at room temperature stirred 1 hour. Make then this mixture by Dowex 50 (H+) Post is so that excessive sodium borohydride changes into boric acid. Make the eluent freeze drying. Make freezing doing Dry thing is dissolved in the aliquot methyl alcohol, makes the filtrate drying. This operation repeats 3 times at least. Then will The dry powder water-soluble solution of aliquot, then freeze drying obtains final compound 6. Collect 125.26mg solid chemical compound 6 (M.W.438.6) (yield: 84.0%).
The 6 couples of resorcinol-HCl of compound that collect and TBA reaction are all aobvious positive.
Embodiment 13
The preparation of compound 7
Make the solid (intermediate 6) of the freeze-drying of 94.26mg (0.34mmol) be dissolved in 5.0ml Distilled water in. The somantadine (0.34mmol) that under slowly stirring, adds then 61.0mg. This mixture is spent the night 4 ℃ of lower slowly stirrings. After adding the sodium borohydride of 40.0mg, make Reactant mixture at room temperature stirred 1 hour. Make then this mixture by Dowex 50 (H+) Post is so that excessive sodium borohydride changes into boric acid. Then with the eluent freeze drying. Then The freeze drying thing is dissolved in the aliquot methyl alcohol, makes the filtrate drying. This operation repeats 3 times at least. With the powder water-soluble solution of aliquot of drying, then freeze drying obtains final compound 7 then. Collect solid chemical compound 7 (the M.W.468.6) (yield: 85.0%) of 135.43mg.
The compound of collecting is all aobvious positive to resorcinol-HCl and TBA reaction.
Embodiment 14
The preparation of compound 8
Make the solid (intermediate 5) of the freeze-drying of 84.0mg (0.34mmol) be dissolved in 5.0ml Distilled water in. The Memantine (0.34mmol) that under slowly stirring, adds then 61.0mg. This mixture is spent the night 4 ℃ of lower slowly stirrings. After adding the sodium borohydride of 40.0mg, make Reactant mixture at room temperature stirred 1 hour. Make then this mixture by Dowex 50 (H+) Post is so that excessive sodium borohydride changes into boric acid. Make the eluent freeze drying. Make freezing doing Dry thing is dissolved in the aliquot methyl alcohol, makes the filtrate drying. This operation repeats 3 times at least. Then will The dry powder water-soluble solution of aliquot, then freeze drying obtains final compound 8. Collect 129.80mg solid chemical compound 8 (M.W.410.5) (yield: 93.0%).
The compound of collecting is all aobvious positive to resorcinol-HCl and TBA reaction. Compound 8 FT-IR spectrum (A) is plotted in Fig. 3, and (B) of contrast is initial intermediate Memantine with it, show with Lower reference value: a) acid amides I bands of a spectrum: 1640cm-1 B) acid amides II bands of a spectrum: 1550cm-1 C) Primary amino radical group: 600-800cm-1，1590-650cm
-1，3330-3380cm
-1 The parahelium group: 700-800cm-1；1615cm
-1；3300cm
-1。
Embodiment 15
The preparation of compound 9
Make the solid (intermediate 6) of the freeze-drying of 94.26mg (0.34mmol) be dissolved in 5.0ml Distilled water in. The Memantine (0.34mmol) that under slowly stirring, adds then 61.0mg. This mixture is spent the night 4 ℃ of lower slowly stirrings. After adding the sodium borohydride of 40.0mg, make Reactant mixture at room temperature stirred 1 hour. Make then this mixture by Dowex 50 (H+) Post is so that excessive sodium borohydride changes into boric acid. Then with the eluent freeze drying. Then The freeze drying thing is dissolved in the aliquot methyl alcohol, makes the filtrate drying. This operation repeats 3 times at least. With the powder water-soluble solution of aliquot of drying, then freeze drying obtains final compound 9 then.
Collect compound 9 (the M.W.440.5) (yield: 90.0%) of 134.79mg.
The 9 couples of resorcinol-HCl of compound that collect and TBA reaction are all aobvious positive.
Embodiment 16
The preparation of compound 10
Make the solid (intermediate 5) of the freeze-drying of 84.0mg (0.34mmol) be dissolved in 5.0ml Distilled water in. The Ribavirin (0.34mmol) that under slowly stirring, adds then 82.7mg. This mixture is spent the night 4 ℃ of lower slowly stirrings. After adding the sodium borohydride of 40.0mg, make Reactant mixture at room temperature stirred 1 hour. Make then this mixture by Dowex 50 (H+) Post is so that excessive sodium borohydride changes into boric acid. Make the eluent freeze drying. Do freezing Dry thing is dissolved in the aliquot methyl alcohol, makes the filtrate drying. This operation repeats 3 times at least. Then will The dry powder water-soluble solution of aliquot, then freeze drying obtains final compound 10. Collect 140.62mg compound 10 (M.W.475.4) (yield: 87.0%). The chemical combination of collecting Thing is aobvious positive to resorcinol-HCl and TBA reaction. The FT-IR spectrum (A) of compound 10 is painted In Fig. 4, (B) of contrast is initial intermediate Ribavirin with it, shows following reference value: a) Acid amides I bands of a spectrum: 1640cm-1 B) acid amides II bands of a spectrum: 1550cm-1 C) primary amino radical group: 600-800cm-1，1590-650cm
-1，3330-3380cm
-1 Parahelium group: 700-800cm-1；
1615cm
-1；3300cm
-1。
Embodiment 17
The preparation of compound 11
Make the intermediate sialic acid of the freeze-drying of 102.5mg (0.34mmol) be dissolved in 50.0ml Methyl alcohol: distilled water (9: 1, in mixture v/v). Under slowly stirring, add 51.442 then The amantadine of mg (0.34mmol). After adding the sodium borohydride of 100.0mg, make reaction mixed Compound is 60 ℃ of lower stirrings 1 hour. Make then this mixture by Dowex 50 (H+) post, In order to make excessive sodium borohydride change into boric acid. Make the eluent freeze drying. With freeze drying Thing is dissolved in the aliquot methyl alcohol, makes the filtrate drying. This operation repeats 5 times at least. To do then The dry solid water-soluble solution of aliquot, then freeze drying obtains final compound 11. Collect total Meter 133.90mg solid chemical compound 11 (M.W.442.5) (yield: 89.0%). Collect Compound is aobvious positive to resorcinol-HCl reaction, and aobvious negative to the TBA reaction.
Embodiment 18
The compounds of this invention suppresses the antiviral activity evaluation of influenza virus A and Type B strain.
Purpose and project
Especially, compare with Memantine with amantadine, having estimated two kinds has the disease-resistant of high value The compound of cytotoxic activity is that compound 1 and compound 8 suppress the external of influenza virus A and Type B Antiviral activity. In this research, used two kinds of influenza virus A types that separate and separated The virus Type B.
Materials and methods
The propagation of 1-influenza virus and titration
Make influenza virus A/H 3N2 (A/ Panama/2007/99), A/H1N1 (A/New Caledonia/20/99) and B (B/ Shandong/7/97) bacterial strain propagate in embryo's egg. Letter It makes strains of influenza viruses be inoculated in 11 days the egg of fertilization by the allantois approach. Make then chicken Egg is collected allantoic fluid 37 ℃ of lower hatchings 3 days, then measures titer.
(plaque assay PA) carries out assay determination by plaque-forming assay. In detail, make The serial dilutions (10 of the virus of each separation-1-10
-11) be inoculated into the fusion in 12 orifice plates In the MDCK of individual layer (Madin-Darby canine kidney) cell. After hatching 1 hour under 37 ℃, remove The virus inoculation thing adds and infects the medium (MEM that contains 10 μ g/ml trypsase, 2% agar (minimum essential medium)). At 37 ℃, 5%CO2After 3 days are hatched in the place, cell monolayer is used 5% glutaraldehyde solution is fixed, remove agar after, with the solution-dyed of 5% carbol fuchsin.
To plaque range estimation counting, this compartment analysis is with every ml plaque forming unit (PFU) (PFU/ml) number represents.
The compound 1 that 2-is relevant with amantadine and Memantine and the antiviral activity of compound 8 Evaluation.
2.1. the preparation of the compound of analyzing
After the drying 72 hours, make compound 1 and 8, and the compound amantadine of contrast Suitably redissolve in sterile distilled water with the concentration of 1000 μ M with Memantine. Then to each test Compound carries out serial dilution take 10 as the basis, and scope is 0.01 μ M to 100 μ M.
2.2. reducing, plaque analyzes (PRA)
Make the fusion individual layer mdck cell (1 * 10 that grows in 12 orifice plates5Cell/ml) with about 50 Each isolated viral of PFU/ml (A/ Panama/2007/99-H3N2, A/ New Caledonia / 20/99-H1N1 and B/ Shandong/7/97) infect. At 37 ℃, 5%CO2Under hatch 1 hour with After making viruses adsorption, remove the virus inoculation thing, this cell monolayer is washed 2 with the MEM culture medium Inferior. In each hole, add covering culture medium (10 μ g/ml trypsase in MEM, 2% fine jade Fat), this covering culture medium contains the analysis of compounds (from 0.01 μ M to 100 μ M) of serial dilution. Double is carried out in test, adds simultaneously the reaction tester that does not contain antiviral compound. Make thus Cell culture is at 37 ℃, 5%CO2Under hatched 3 days. Then, make cell monolayer with 5% penta 2 Aldehyde solution is fixed, and at room temperature hatches at least 3 hours again, penetrates in the agar with promotion. After removing agar, make cell monolayer dyeing with 5% carbolfuchsin solution. Range estimation counting plaque, Calculate the degree that plaque suppresses with respect to the tester that does not contain test compounds. Measure thus phase Plaque decreased number 50% necessary compound for the tester that does not contain The compounds of this invention Concentration.
The result
The antiviral activity of compound 1 and compound 8 and control compound amantadine and U.S. The antiviral activity of Buddha's warrior attendant is to reduce analysis (PRA) by plaque to estimate with mdck cell . The results are shown in table 1.
8 pairs of A type influenza viruses (A/H3N2 and A/H1N1) that suppress to analyze demonstrate compound Antiviral activity than the big 10 times of magnitudes of Memantine. In the test concentrations scope, compound 8 Demonstrate the antiviral activity (table 1) that suppresses the Type B influenza virus.
Table 1
Conclusion:
The data that provide show that compound 1 than amantadine, has improved inhibition Type B influenza The antiviral activity of virus. Need to remember that amantadine is to exist only in A type stream by blocking-up Influenza Virus and be not present in the M2 albumen onset of the ion channel in the Type B influenza virus, therefore This may be a kind of new mechanism of action, and this may be the target of future studies.
Compound 8 demonstrates the antiviral work of the inhibition A type influenza virus higher 10 times than Memantine The property.
Claims (23)
1. the compound of structural formula (I), with and the C of straight or branched
1-4The carboxyl list-or many-ester, additive salt, solvate, the enantiomorph of fractionation and the diastereomer of purifying;
Wherein:
X represents-CH
2-,-O-,-CHF-,-CF
2-;
The C of singly-bound or two key shacks
2-C
3
R
1Expression-OH, halogen or-the B/ part, condition is the C that has two key shacks
2-C
3The time R
1Do not exist; And
R
2Be-OH ,-O-CH (C
2H
5)
2,-NH
2,-NHC (NH) NH
2Or-NH-OH; And
R
3Be-NH
2,-NHCO-CH
3Or-NH-CO-CH
2-OH; And
R
4Expression-CHOH-CHOH-CH
2-OH ,-CHOH-CH
2-B/ part ,-CH
2-B/ part or-the B/ part; And
Wherein-B/ partly represents:
-B1/ part-B2/ part
Wherein:
R
5Expression connection functional group-NH-,-CH
2-NH-,-CH (CH
3)-NH-,-NH-CH (CH
3)-NH-,-C (CH
3)
2-CH
2-NH-,-NH-CO-CH
2-O-CH
2-CH
2-NH-or
And
R
6Be-H ,-CH
3Or-C
2H
5And
R
7Be-H ,-CH
3Or-C
2H
5And
R
8Expression connect functional group-NH-,-CO-NH-or-C (NH)-NH-.
2. the compound of claim 1, it is to be selected from following C
7Or C
8The compound of list-B/ part-replacement:
Compound 1
Compound 2
Compound 3
Compound 4
Compound 5
Compound 6
Compound 7
Compound 8
Compound 9
Compound 10.
3. the compound of claim 1, it is to be selected from following C
2The compound of list-B/ part-replacement:
4. the compound of claim 1, it is C
2And C
8The compound of two-B1/ part-replacement:
5. the compound of claim 1, it is C
2And C
8The compound of two-B2/ part-replacement:
6. the compound of claim 1, it is to be selected from following C
2And C
8Have different-B1/ part or-B2/ part-disubstituted compound:
7. pharmaceutical composition, it is included in each described compound of claim 1 to 6 or its mixture in the pharmaceutically acceptable carrier.
8. treatment is by erythrocyte agglutination element (HA) and/or neuraminidase (NA) and/or contain M
2The method of the virus infection that proteic virus causes, this method comprise each the compound of claim 1 to 6 of giving the host's administering therapeutic significant quantity that needs this treatment by any suitable route of administration.
9. the method for claim 8, wherein said infection are that virus or its mutant by influenza virus A and Type B causes.
10. treatment is by the polymerase-mediated disease of ARNm and ARN or the method for illness, and this method comprises each the compound of inhibition ARNm-guanidine radicals transferring enzyme of claim 1 to 6 of giving the host's administering therapeutic significant quantity that needs this treatment by any suitable route of administration.
11. being hepatitis C viruss (HVC), the method for claim 10, wherein said ARNm, ARN polysaccharase and the transferase mediated infection of ARNm-guanidine radicals infect.
12. the method for claim 9, it further provides and makes described compound and the another kind of effectively inhibition influenza virus A of treatment significant quantity and the compound combined administration of Type B or its mutant.
13. the method for claim 12, the compound of wherein said effective inhibition influenza virus A and Type B or its mutant is zanamivir and/or oseltamivir.
14. the method for claim 11, it further provides and makes described compound and the another kind of effectively inhibition hepatitis C virus (HVC) of treatment significant quantity or the compound combined administration of its mutant.
15. the method for claim 14, the compound of wherein said effective inhibition HVC is interferon alpha or polyoxyethylene glycol interferon alpha, makes up separately or with ribavirin.
16. each compound of the claim 1 to 6 that is used for the treatment of.
17. the compound of claim 16, wherein said treatment are HA or NA or M in Mammals
2The treatment of protein mediated virus infection.
18. the compound of claim 16, wherein said treatment are the treatments of the transferase mediated HVC of ARNm, ARN polysaccharase and ARNm-guanidine radicals in Mammals.
19. the compound of claim 17 or 18, wherein said virus infection are to be caused by any of A type influenza virus or Type B influenza virus, perhaps hypotype and different strain and the mutant by them causes.
20. each compound of claim 1 to 6 is used at Mammals treatment HA or NA or M in preparation
2Purposes in the protein mediated medicine for treating viral infections.
21. each compound of claim 1 to 6 is used for treating purposes in the transferase mediated medicine for treating viral infections of ARNm, ARN polysaccharase and ARNm-guanidine radicals Mammals in preparation.
22. the purposes of claim 20 or 21, wherein said virus infection are caused with different strain and mutant by A type influenza virus or Type B influenza virus or their hypotype.
23. the purposes of claim 22, wherein said virus infection are caused by any of A or Type B influenza virus or hepatitis C virus or their hypotype and different strain and mutant.
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| ITMI20070093 ITMI20070093A1 (en) | 2007-01-23 | 2007-01-23 | ANTIVIRAL COMPOUNDS |
| ITMI2007A000093 | 2007-01-23 | ||
| ITMI2007A002212 | 2007-11-22 | ||
| ITMI20072212 ITMI20072212A1 (en) | 2007-11-22 | 2007-11-22 | ANTIVIRAL COMPOUNDS |
| PCT/EP2008/050703 WO2008090151A1 (en) | 2007-01-23 | 2008-01-22 | Antiviral compounds |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101622265A true CN101622265A (en) | 2010-01-06 |
| CN101622265B CN101622265B (en) | 2015-06-10 |
Family
ID=40273623
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200880006407.8A Expired - Fee Related CN101622265B (en) | 2007-01-23 | 2008-01-22 | Antiviral compounds |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN101622265B (en) |
| IT (1) | ITMI20070093A1 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103509066A (en) * | 2012-06-25 | 2014-01-15 | 沈阳药科大学 | Lipid derivative containing sialic acid fragment and application thereof |
| CN104031097A (en) * | 2013-03-04 | 2014-09-10 | 沈阳药科大学 | Lipid derivative containing sialic acid group and application thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992006691A1 (en) * | 1990-10-19 | 1992-04-30 | Biota Scientific Management Pty. Ltd. | Anti-viral compounds that bind the active site of influenza neuramidase and display in vivo activity against orthomyxovirus and paramyxovirus |
| US6555698B1 (en) * | 1998-11-17 | 2003-04-29 | Tropix, Inc. | Chemiluminescent substrates for neuraminidase, assays for detection of neuraminidase and kits therefor |
-
2007
- 2007-01-23 IT ITMI20070093 patent/ITMI20070093A1/en unknown
-
2008
- 2008-01-22 CN CN200880006407.8A patent/CN101622265B/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103509066A (en) * | 2012-06-25 | 2014-01-15 | 沈阳药科大学 | Lipid derivative containing sialic acid fragment and application thereof |
| CN103509066B (en) * | 2012-06-25 | 2016-03-30 | 沈阳药科大学 | A kind of lipid derivate and application thereof containing sialic acid fragment |
| CN104031097A (en) * | 2013-03-04 | 2014-09-10 | 沈阳药科大学 | Lipid derivative containing sialic acid group and application thereof |
| CN104031097B (en) * | 2013-03-04 | 2016-12-28 | 沈阳药科大学 | A kind of lipid derivate containing sialic acids groups and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101622265B (en) | 2015-06-10 |
| ITMI20070093A1 (en) | 2008-07-24 |
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