CN101627019A - Bicyclic pyrimidinones with and use - Google Patents

Bicyclic pyrimidinones with and use Download PDF

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CN101627019A
CN101627019A CN200780051562.7A CN200780051562A CN101627019A CN 101627019 A CN101627019 A CN 101627019A CN 200780051562 A CN200780051562 A CN 200780051562A CN 101627019 A CN101627019 A CN 101627019A
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hydroxyl
alkyl
oxo
pyrimidine
300mhz
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CN101627019B (en
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埃里克·戴尔·琼斯
乔纳森·艾伦·维克多·科茨
戴维·伊恩·罗兹
约翰·约瑟夫·戴德曼
尼古拉斯·安德鲁·范德格拉夫
莉萨·简·温菲尔德
内尔纳·天通
威廉·伊萨
尼尔·乔依
凯瑟琳·麦克法兰
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Tali Digital Ltd
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Avexa Ltd
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Abstract

The invention provides a kind of compound or its medicinal derivative, salt or prodrug of Formula I.The method of a kind of treatment or prevention curee virus infection is provided in addition, has comprised compound or its medicinal derivative, salt or the prodrug of the Formula I that gives described curee's significant quantity.The pharmaceutical composition or the medicine of the compound that comprises Formula I also are provided.

Description

Bicyclic pyrimidinones with and use
Technical field
The present invention relates to be used for the treatment of virus infection, especially the novel bicyclic pyrimidinone compound that infects of HIV with and analogue.
Background technology
The retrovirus that is called " human immunodeficiency virus " or " HIV " is a cause of disease (etiological agent) of progressively destroying immune complex disease.This disease is called acquired immune deficiency syndrome (AIDS) or AIDS.As in December, 2005, estimate that the whole world 40,000,000 people suffer from HIV and the annual death that three million peoples take place to surpass.
The characteristics that retrovirus duplicates comprise the viral genome reverse transcription enter among provirus (provirus) DNA with and be integrated in the host cell gene group.It is needed that these steps are that HIV duplicates, and respectively by encoding viral enzyme, reversed transcriptive enzyme and integrase mediated.
HIV infects and follows following approach: virion is incorporated into cell surface receptor and co-receptor, causes the fusion of virion and cell.The inclusion of virus is released in the tenuigenin, and the genomic reverse transcription of HIV takes place in this tenuigenin.By series of steps, produced double-stranded proviral DNA copy.Proviral DNA be transported to be called integrate before (PIC) nucleus in the complex body of (it comprises intergrase and other virus and possible cell protein) of complex body (pre-integration complex).In case in nucleus, then, proviral DNA is incorporated in the host cell gene group by means of the effect of intergrase.In case be integrated, then can take place virus genomicly to transcribe and translate, cause producing viral protein and new virus rna gene group.These albumen and genome are assemblied on the cell surface, and depend on cell type, may be assembled in the interior membrane compartment of other cells.Make then the particle of assembling begin from cell grow out and during this process or after this process soon, the effect maturation by virus protease is the infected by HIV particle.
The provirus genome needs the effect of intergrase to the integration of host cell gene group, its with at least three steps, may four steps carry out this process.First step relates to viral genome is assembled in the stable nucleoprotein complex body, secondly, to processing the staggered end that has free 3 ' OH residue with generation from two Nucleotide of genomic 3 ' end, and the 3rd, these ends are transferred in the host cell gene group.Final step relates to the gap filling and the reparation of inserting the site in host genome.For whether intergrase carries out this final step or do not carry out this final step by the cytothesis enzyme, still there are some guesses.
At present, can treat HIV with the multiple inhibitor of selling and infect, wherein inhibitor target reversed transcriptive enzyme, proteolytic enzyme, or enter cell.Known, can treat AIDS and similar disease effectively with the combined therapy HIV infection of these medicines or these medicines.The shortcoming of inhibitor comprises the rapid appearance of resistance and many side effects and the incidence of increase at present, therefore needs the proteinic new inhibitor of target such as intergrase.
Summary of the invention
Aspect first, the invention provides a kind of compound or its medicinal derivative, salt or prodrug of Formula I,
Wherein:
A condenses in the monocycle that contains azo-cycle or dicyclo (two rings) aromatics or heteroaromatic moiety;
X is selected from the group of being made up of O and S;
Y is selected from the group of being made up of O and S;
R 1Be 0-3 substituting group, its each be independently selected from by CN, C 1-10Alkyl, C 2-10Alkenyl, C 1-10Alkyl PO 3H 2,-O-C 1-10Alkyl, C 1-10Alkyl NR 3R 4,-O-C 1-10Alkyl NR 3R 4, halogen (halo), NR 3R 4, alkaryl, miscellaneous alkyl aryl, aryl, heteroaryl ,-O-alkaryl, SO 2NR 3R 4The group of forming;
R 3And R 4Be selected from independently of one another by hydrogen, CO 2C 1-4Alkyl, C (O) C 1-4Alkyl, C 1-10Alkyl, C 1-10NR 5R 6,-NH (CO) is NHC (CO) 1-4The group that alkyl is formed; Or R 3And R 4Form 5-7 unit heterocycle with the nitrogen that connects, this heterocycle comprises 0 to 2 other heteroatoms that is selected from N, O or S, and wherein S can be in S, S (O) or S (O) 2Oxidation state, and wherein said heterocycle at carbon or nitrogen-atoms place alternatively by one or more halogen, aryl, C (O) C of being selected from 1-4Alkyl, SO 2C 1-4Alkyl, SO 2H, C 1-4Alkyl, CO 2H, CO 2C 1-4Alkyl, NR 5R 6, C 1-4Alkyl NR 5R 6Substituting group replace;
R 5And R 6Be selected from independently of one another by H and C 1-4The group that alkyl is formed, or R 5And R 6Form 5-7 unit heterocycle with the nitrogen that is connected, this heterocycle comprises 0 to 2 other heteroatoms that is selected from N, O or S, and wherein S can be in (being) S, S (O) or S (O) 2Oxidation state, and wherein said heterocycle at carbon atom or nitrogen-atoms place alternatively by one or more halogen and C of being selected from 1-4The substituting group of alkyl replaces;
Work as R 1Be alkaryl or-during the O-alkaryl, the substituent aromatic yl group of described alkaryl is selected from C alternatively 1-10Alkyl ,-O-C 1-10Alkyl, C 1-10Alkyl NR 3R 4,-O-C 1-10Alkyl NR 3R 4, halogen, NR 3R 4, alkaryl ,-O-alkaryl, SO 2NR 3R 4Substituting group replace;
B do not exist or-C (O)-;
C do not exist or be selected from by-O-,-NH-and-group of NH-NH-C (O)-form;
R 2Be selected from by heteroaryl, heterocyclic radical and R 7The group of forming;
R 7Be selected from H, alkaryl and C 1-10Alkyl;
Condition is if R 2Be R 7, then B and C must exist.
Aspect second, the invention provides the method for virus infection among a kind of treatment or the prevention curee, comprise compound or its medicinal derivative, salt or the prodrug of the chemical formula (I) that gives described curee's significant quantity.
Aspect the 3rd, provide the compound of Formula I or its medicinal derivative, salt or prodrug to be used for the treatment of or to prevent the application in the medicine for treating viral infections among the curee in preparation.
Aspect the 4th, the invention provides pharmaceutical composition, this pharmaceutical composition comprises according to the compound of first aspect and pharmaceutical carrier, thinner or vehicle.
Description of drawings
Fig. 1 shows the activity table of compound (comprising according to compound of the present invention) in the chain transfer that utilizes wild-type and sudden change hiv integrase is measured.Mutant enzyme comprises sudden change Q148K and has resistance for the integrase inhibitor of announcing such as S-1360, Raltegravir (MerckMK-0518) and GS9137 (Gilead GS-9137).
Embodiment
Aspect first, the invention provides a kind of compound or its medicinal derivative, salt or prodrug of Formula I,
Figure G2007800515627D00041
Wherein:
A condenses in the monocycle that contains azo-cycle or bicyclic aromatic or heteroaromatic moiety;
X is selected from the group of being made up of O and S;
Y is selected from the group of being made up of O and S;
R 1Be 0-3 substituting group, it is selected from independently of one another by CN, C 1-10Alkyl, C 2-10Alkenyl, C 1-10Alkyl PO 3H 2,-O-C 1-10Alkyl, C 1-10Alkyl NR 3R 4,-O-C 1-10Alkyl NR 3R 4, halogen, NR 3R 4, alkaryl, miscellaneous alkyl aryl, aryl, heteroaryl ,-O-alkaryl, SO 2NR 3R 4The group of forming;
R 3And R 4Be selected from independently of one another by hydrogen, CO 2C 1-4Alkyl, C (O) C 1-4Alkyl, C 1-10Alkyl, C 1-10NR 5R 6,-NH (CO) is NHC (CO) 1-4The group that alkyl is formed; Or R 3And R 4Form 5-7 unit heterocycle with the nitrogen that is connected, this heterocycle comprises 0 to 2 other heteroatoms that is selected from N, O or S, and wherein S can be in S, S (O) or S (O) 2Oxidation state, and wherein said heterocycle at carbon atom or nitrogen-atoms place alternatively by one or more halogen, aryl, C (O) C of being selected from 1-4Alkyl, SO 2C 1-4Alkyl, SO 2H, C 1-4Alkyl, CO 2H, CO 2C 1-4Alkyl, NR 5R 6, C 1-4Alkyl NR 5R 6Substituting group replace;
R 5And R 6Be selected from independently of one another by H and C 1-4The group that alkyl is formed, or R 5And R 6Form 5-7 unit heterocycle with the nitrogen that is connected, this heterocycle comprises 0 to 2 other heteroatoms that is selected from N, O or S, and wherein S can be in (being) S, S (O) or S (O) 2Oxidation state, and wherein said heterocycle at carbon or nitrogen-atoms place alternatively by one or more halogen and C of being selected from 1-4The substituting group of alkyl replaces;
Work as R 1Be alkaryl or-during the O-alkaryl, the substituent aromatic yl group of described alkaryl is selected from C alternatively 1-10Alkyl ,-O-C 1-10Alkyl, C 1-10Alkyl NR 3R 4,-O-C 1-10Alkyl NR 3R 4, halogen, NR 3R 4, alkaryl ,-O-alkaryl, SO 2NR 3R 4Substituting group replace;
B do not exist or-C (O)-;
C do not exist or be selected from by-O-,-NH-and-group of NH-NH-C (O)-form;
R 2Be selected from by heteroaryl, heterocyclic radical and R 7The group of forming;
R 7Be selected from H, alkaryl and C 1-10Alkyl;
Condition is if R 2Be R 7, then B and C must exist.
In a kind of preferred form, R 2Be heteroaryl or heterocyclic radical.More preferably, R 2Replaced by aryl or alkaryl.
In a kind of preferred form, C 2-10Alkenyl is an allyl group.
In a kind of preferred form, the compound of Formula I is selected from the group of being made up of the compound of Formulae II, III, IV, V and VI:
Figure G2007800515627D00061
In the compound of Formulae II I, IV and V, Z is O, S or NR 8, R wherein 8Be H, C 1-10Alkyl, C 1-10Alkyl NR 3R 4, alkaryl, miscellaneous alkyl aryl, aryl and heteroaryl.
In further preferred form, the compound of Formula I is the compound of chemical formula VII:
Figure G2007800515627D00062
In a kind of preferred form, NR 3R 4It is morpholine.
In a kind of preferred form, R 7Be luorobenzyl, more preferably 4-luorobenzyl.
In the preferred form of another kind, R 7Be dichloro benzyl, more preferably 3, the 4-dichloro benzyl.
Preferably, heteroaryl is selected from the group of being made up of tetrazolium, triazole, pyrazoles, imidazoles, oxazole, oxadiazole, thiazole, thiadiazoles.
Preferably, this compound is selected from:
7-bromo-3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
3-hydroxyl-7-methyl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
3-hydroxyl-7-methyl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 3,4-two chloro-benzyl acid amides
7-chloro-3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 3,4-two chloro-benzyl acid amides
7-chloro-3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
7-(1,1-dioxide-isothiazolidine-2-yl)-3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
7-(1,1-dioxide-[1,2] thiazan-2-yl)-3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
3-hydroxyl-4-oxo-7-piperidines-1-base-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
3-hydroxyl-7-morpholine-4-base-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
3-hydroxyl-7-(4-methyl-piperazine-1-yl)-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
4-[2-(4-fluoro-benzylamino formyl radical)-3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidin-7-yl]-piperazine-1-carboxylic acid tert-butyl ester
7-(2-dimethylamino-ethylamino)-3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
3-hydroxyl-4-oxo-7-(2-tetramethyleneimine-1-base-ethylamino)-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
N-[2-(4-fluoro-benzylamino formyl radical)-3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidin-7-yl]-N ', N '-dimethyl-oxalamide (oxamide)
3-hydroxyl-7-(2-morpholine-4-base-ethylamino)-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
9-dimethylamino-3-hydroxyl-7-morpholine-4-base-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
9-ethyl-3-hydroxyl-7-morpholine-4-base-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
3-hydroxyl-7-morpholine-4-base-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 3-chloro-2-fluoro-benzyl acid amides
7-(3-chloro-2-fluoro-benzyl)-3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid (the 5-methyl-[1,3,4] oxadiazole-2-ylmethyls)-acid amides
7-(4-fluoro-benzyl)-3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid benzyl acid amides
7-(4-fluoro-benzyl)-3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
7-(3-chloro-2-fluoro-benzyl)-3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid (1H-tetrazolium-5-yl)-acid amides
7-(3-chloro-2-fluoro-benzyl)-3-hydroxyl-2-(2H-tetrazolium-5-yl)-pyrido [1,2-α] pyrimidin-4-one
7-(3-chloro-2-fluoro-benzyl)-3-hydroxyl-2-(2H-[1,2,4] triazole-3-carbonyl)-pyrido [1,2-α] pyrimidin-4-one
7-(3-chloro-2-fluoro-benzyl)-3-hydroxyl-2-(1H-tetrazolium-5-carbonyl)-pyrido [1,2-α] pyrimidin-4-one
3-hydroxyl-8-morpholine-4-base-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
7-(3-chloro-2-fluoro-benzyl)-3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid
7-(4-fluoro-benzyl)-3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid
3-hydroxyl-6-morpholine-4-base-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
3-hydroxyl-9-morpholine-4-base-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
7-bromo-3-hydroxyl-4-sulfo--4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
3-sulfydryl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid N '-[2-(4-fluoro-phenyl)-ethanoyl]-hydrazides
2-[5-(4-fluoro-benzyl)-[1,3,4] oxadiazole-2-yl]-3-hydroxyl-pyrido [1,2-a] pyrimidin-4-one
3-hydroxyl-7-(morpholine-4-alkylsulfonyl)-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
3-hydroxyl-4-oxo-7-sulphonamide-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
7-dimethylamino sulphonyl-3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
3-hydroxyl-4-oxo-7-tetramethyleneimine-1-ylmethyl-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
7-dimethylaminomethyl-3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
Aspect second, the invention provides the method for virus infection among a kind of treatment or the prevention curee, this method comprises compound or its medicinal derivative, salt or the prodrug of the chemical formula (I) that gives described curee's significant quantity.
Aspect the 3rd, provide the compound of Formula I or its medicinal derivative, salt or prodrug to be used for the treatment of or to prevent the application in the medicine for treating viral infections among the curee in preparation.
Preferably, the virus infection of second and the 3rd aspect is that HIV or SIV infect.
More preferably, HIV or SIV infect and comprise the virus strain that has resistance for other integrase inhibitors.Even more preferably, this virus strain comprises the hiv integrase with Q148K sudden change.Some compound of the present invention has demonstrated with respect to hiv integrase (Q148K) has surprising more high reactivity, and wherein hiv integrase is for the integrase inhibitor such as S-1360, Raltegravir (Merck MK-0518) and the GS9137 (Gilead GS-9137 has resistance) that announce.Show the compound (referring to Fig. 1) that highly active especially compound is embodiment 6.13 with respect to Q148K.
An example of the reference that the resistance of Q148K distributes can be at 14th CROI (Conference on Retroviral and Opportunistic Infections), Los Angeles, February 27th 2007 from John Wai, Merck Research Labs finds among ' Nextgeneration inhibitors of HIV-I Integrase Strand Transfer:Structuraldiversity and resistance the profiles.
As employed in this article, term " halogen " or " halogen " are meant fluorine (fluorine-based), chlorine (chloro), bromine (bromo) or iodine (iodo).
As employed in this article, separately or at compound term such as NH (alkyl) or N (alkyl) 2The middle term " alkyl " that uses is meant unit price straight or branched hydrocarbyl group, and in appropriate circumstances, it has 1 to 3,1 to 6 or 1 to 10 carbon atom.For example, Shi Yi alkyl group includes but not limited to methyl, ethyl, propyl group, sec.-propyl, normal-butyl, sec-butyl, the tertiary butyl, amyl group, 2-methyl butyl, 3-methyl butyl, n-hexyl, 2-, 3-or 4-methyl amyl, 2-ethyl-butyl, n-hexyl or 2-, 3-, 4-or 5-methyl amyl.
As employed in this article, term " alkenyl " is meant the straight or branched hydrocarbyl group that has one or more pairs of keys between carbon atom.Suitable kiki alkenyl group includes but not limited to vinyl, allyl group, propenyl, pseudoallyl, butenyl, pentenyl and hexenyl.
As employed in this article, term " cycloalkyl " is meant cyclic hydrocarbon group.Suitable group of naphthene base includes but not limited to cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.
As employed in this article, term " aryl " is meant C 6-C 10Aromatic hydrocarbon group, for example phenyl or naphthyl.
Term " alkaryl " comprises, for example, and benzyl.
When separately or when using in the compound term, term " heterocycle " comprises monocycle, encircles more, condenses or the conjugated hydrocarbons residue, preferred C 3-6, wherein one or more carbon atoms (and under suitable situation, it is connected with hydrogen atom) are replaced by heteroatoms, and making provides non-aromatic moieties.Key between the atom can be saturated or unsaturated.Suitable heteroatoms comprises O, N and S.Under the substituted situation of two or more carbon atoms, it can replace by two or more identical heteroatomss or by different heteroatomss.The suitable example of heterocyclic group can comprise that pyrrolidyl, piperidyl, piperazinyl, morpholino, quinolyl, isoquinolyl, thiomorpholine are for, alkyl dioxin, 2,2 '-dimethyl-[1,3]-dioxolanyl, tetrahydrofuran base, THP trtrahydropyranyl, Pyrrolidine base etc.
Term " heteroaryl " comprises 5 yuan or 6 yuan of assorted aromatic nucleus, and it comprises the one or more heteroatomss that are selected from O, N and S.The suitable example of heteroaryl groups comprises furyl, thiophenyl, tetrazyl, 1,2,3-triazoles base, 1,2,4-triazolyl, imidazolyl, pyrazolyl, pyridyl, pyrimidyl, oxazolyl, oxadiazole base, thiazolyl, thiadiazolyl group etc.Assorted aromatic nucleus can be fused to 5 yuan or 6 yuan of aromatic nucleus or assorted aromatic nucleus to form for example cumarone of dicyclo aromatic ring system.
Except as otherwise noted, otherwise each alkyl, cycloalkyl, alkaryl, aryl, heterocyclic radical or heteroaryl groups can be alternatively by C 1-C 3Alkyl, C 3-C 6Cycloalkyl, C 6Aryl, heterocyclic radical, heteroaryl, C 1-C 3Alkyl OH, alkaryl, OH, OC 1-C 3Alkyl, halogen, CN, NO 2, CO 2H, CO 2C 1-C 3Alkyl, CONH 2, CONH (C 1-C 3Alkyl), CON (C 1-C 3Alkyl) 2, trifluoromethyl, NH 2, NH (C 1-C 3Alkyl) or N (C 1-C 3Alkyl) 2In one or more replacements.For example, the aromatic yl group of optional replacement can be 4-aminomethyl phenyl or 4-hydroxyl phenylic group, and the alkyl group of optional replacement can be 2-hydroxyethyl, trifluoromethyl or difluoromethyl.Each optional alkyl, cycloalkyl, alkaryl, aryl, heterocyclic radical or heteroaryl substituting group can also be by optional replacements.
Optional substituent example also comprises suitable nitrogen-protecting group group (referring to " ProtectiveGroups in Organic Synthesis " Theodora Greene and Peter Wuts, thirdedition, Wiley Interscience, 1999).
The salt of the compound of Formula I is preferably medicinal, but should understand, non-pharmaceutical salts also falls within the scope of the invention, because they can be used as intermediate in the preparation of pharmaceutical salts.
Term " medicinal derivative " can comprise any pharmaceutical salts, hydrate or prodrug, or any other compound, and it can provide compound or its anti-microbial activity metabolite or the resistates of (directly or indirectly) Formula I after giving the curee.
Suitable pharmaceutical salts includes but not limited to the salt example hydrochloric acid of medicinal mineral acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, boric acid, thionamic acid, and hydrobromic salt, or pharmaceutically acceptable organic acid such as acetate, propionic acid, butyric acid, tartrate, toxilic acid, hydroxymaleic acid, fumaric acid, oxysuccinic acid, citric acid, lactic acid, glactaric acid, gluconic acid, phenylformic acid, succsinic acid, oxalic acid, toluylic acid, methylsulfonic acid, toluene sulfonic acide, Phenylsulfonic acid, Whitfield's ointment, Sulphanilic Acid, aspartic acid, L-glutamic acid, ethylenediamine tetraacetic acid (EDTA), stearic acid, palmitinic acid, oleic acid, lauric acid, pantothenic acid, tannic acid, the salt of xitix and valeric acid.
Subsalt includes but not limited to that those are by medicinal cation, subsalt as sodium, potassium, lithium, calcium, magnesium, zinc, ammonium, alkylammonium formation, as the salt that forms by triethylamine, the salt that alkoxyl group ammonium such as those are formed by thanomin and by quadrol, choline or amino acid, the salt that forms as arginine, Methionin or Histidine.About the general information of the type of pharmaceutical salts and their formation be it is known to the person skilled in the art that and as textbook, as " Handbook of Pharmaceutical salts " P.H.Stahl, C.G.Wermuth, 1 StEdition, 2002, described in the Wiley-VCH.
The nitrogen-containing group of alkalescence (basic) can be in addition quaternized with such preparation, as elementary alkyl halide, as methyl, ethyl, propyl group and butyl muriate, bromide and iodide; Sulfuric acid dialkyl such as methyl-sulfate and ethyl sulfate; Or the like.
Oh group can be with comprising the low-grade alkyl carboxylic acid, and as acetate and 2, the esterification or with comprising alkylsulphonic acid in addition of the group of 2-neopentanoic acid is as the group sulfonation in addition of methylsulphonic acid (referring to for example, the compound of embodiment 15.10).
The present invention also comprises pharmaceutical composition, and it comprises the prodrug of the compound of Formula I.The present invention also comprises by the prodrug of the compound that gives Formula I and treating in the curee or the method for prophylaxis of viral infections.Compound with Formula I of free amine group, amido, hydroxyl or carboxyl can be converted to prodrug.
Prodrug comprises such compound, and wherein amino-acid residue or two or more (for example, two, three or four) polypeptide chains of amino-acid residue are covalently attached to free amine group, hydroxyl and the hydroxy-acid group of the compound of Formula I.Amino-acid residue comprises 20 kinds of naturally occurring amino acid (representing with three letter characters usually) and comprises 4-oxyproline, hydroxylysine, desmosine (demosine), isodesmosine, 3-Methyl histidine, norvaline, Beta-alanine, γ-An Jidingsuan, citrulline, homocysteine, homoserine (homoserine), ornithine and methionine(Met) sulfone.Prodrug also comprises such compound, and wherein carbonic ether, carbamate, acid amides and alkyl ester are covalently attached to the above-mentioned substituting group of Formula I by carbonyl carbon prodrug side chain.Prodrug also comprises phosphoric acid salt (ester) derivative (as sour, sour salt or ester) of the compound of Formula I, and it is connected in the free hydroxyl group of the compound of Formula I by phosphorus oxygen key.
Should understand that also the compound of Formula I can have asymmetric center, therefore can be to exist more than a kind of stereoisomeric forms in any ratio.Therefore the invention still further relates at one or more asymmetric centers and have the compound of pure isomeric forms basically, for example, greater than about 90%ee, according to appointment 95% or 97%ee or greater than 99%ee, with and composition thereof, comprise racemic mixture.Such isomer can prepare by asymmetric synthesis, for example utilizes chiral intermediate or passes through chiral separation.
In fourth aspect, the invention provides and comprise according to the compound of first aspect and the pharmaceutical composition of pharmaceutical carrier, thinner or vehicle.
Composition of the present invention can comprise other treatment agent as described below, and can be for example according to those technology as knowing in pharmaceutical dosage form (pharmaceutical formulation) field, prepared by the medicated premix (for example, vehicle, tackiness agent, sanitas, stablizer, sweetener etc.) that adopts conventional solid or liquid vehicle or thinner and be suitable for desired mode of administration type.
Can give compound of the present invention by any suitable mode, for example, outside the gi tract, as by subcutaneous, intravenously, intramuscular or intracisternal injection or infusion techniques (for example, as aseptic injection water or non-aqueous solution or suspension).
Pharmaceutical dosage form comprises that those are used for the formulation that oral, rectum, nose, part (comprising cheek and hypogloeeis), vagina or gi tract outer (comprising intramuscular, subcutaneous and intravenously) gave or had the form that the inhalation of being adapted to pass through or insufflation give.The form that compound of the present invention and conventional adjuvant, carrier or thinner can be formulated into pharmaceutical composition thus with and unitary dose, and having such form makes and can be used as solid, as tablet or capsule through filling, or liquid such as solution, suspension, emulsion, elixir or be filled with the capsule of identical component, all be used to orally use, form with suppository is used for rectum and gives; Or have the form of aseptic injectable solution, be used for gi tract outer (comprising subcutaneous) and use.
Except the primate such as the mankind, the method according to this invention can also be treated various other Mammalss.For example, can treat and include but not limited to milk cow, sheep, goat, horse, dog, cat, cavy, the Mammals of rat or other oxen, sheep, equine species, Canis animals, feline, rodent or mouse species.Yet this method can also be used for other species, as fowl species (for example, chicken).
Curee with the aforesaid method treatment is a Mammals, and it includes but not limited to milk cow, sheep, goat, horse, dog, cat, cavy, rat or other oxen, sheep, equine species, Canis animals, feline, rodent or mouse species, and preferred people's (sex).
Term " significant quantity " is meant the amount with the theme composition of induced tissue, system, animal or human's biology or medical response, and this reaction is that scientific research personnel, animal doctor, doctor or other clinicists look for.
As at the treatment virus infection, and especially technician in the HIV field of infecting will understand, term " treatment " the definiteness virus infection that differs is cured fully.Term " treatment " comprises any minimizing of quantity of viruses and/or the inhibition of duplicating in the curee who receives treatment.
As employed in this article, term " composition " is intended to contain a kind of product, and it comprises the appointment composition of specified amount, and any directly or indirectly by the product of the combination results of the appointment composition of specified amount.Term " pharmaceutical " is meant that carrier, thinner or vehicle must and not be harmful to its recipient with other component compatibility of formulation.
It is to point to need the individuality of treatment that compound of the present invention is provided that term " gives " compound.
The pharmaceutical composition that is used to give compound of the present invention can be provided with dosage unit form easily and can be prepared by any well-known method of pharmacy field.All methods may further comprise the steps: active ingredient is combined with carrier, and it constitutes one or more ancillary components.Usually, combine by equal even active ingredient and liquid vehicles or ground solid carrier or both of closely making, then, if necessary, making product shaping is that desired formulation is come pharmaceutical compositions.In pharmaceutical composition, the active target compound that comprises q.s is to produce desired effects to lysis or state.As employed in this article, term " composition " is intended to contain a kind of product, and it comprises the appointment composition of specified amount, and any directly or indirectly by the product of the combination results of the appointment composition of specified amount.
It is moisture or contain the form of oil suspension that pharmaceutical composition can have aseptic injection.Can be according to known technology and utilize those the suitable dispersion agents above mentioned or wetting agent and suspension agent to prepare this suspension.Aseptic injection preparation (preparation) can also be aseptic injectable solution or the suspension in non-toxicity gi tract external application thinner or solvent, for example as the solution in 1,3 butylene glycol.(carrier vehicles) He in the solvent has water, Ringer's solution and isotonic sodium chlorrde solution at the acceptable inert matter that can adopt.In addition, aseptic fixed oil is conventionally used as solvent or suspension medium.For this purpose, can adopt the fixed oil of any gentleness, it comprises synthetic direactive glyceride or triglyceride.In addition, lipid acid such as oleic acid can be used for the preparation of injection.
Pharmaceutical composition of the present invention and method may further include the other treatment active compound, and it is generally used for treating above-mentioned pathological state.According to traditional pharmacy principle, those of ordinary skills can select to be used for the suitable medicament of conjoint therapy.The combination of therapeutical agent can act synergistically to realize treatment or to prevent above-mentioned various obstacle.Utilize this mode, can obtain therapeutic efficiency, thereby reduce the potentiality of harmful side effect with the more low dosage of every kind of medicament (agent).
When being used in combination the other treatment agent with compound of the present invention, their consumption for example can be as that mentioned in Physician Desk Reference (PDR) or as otherwise determined by those of ordinary skills.
Need in the illness of HIV inhibition or hiv integrase inhibition in treatment or prevention, suitable dosage level will be generally about 0.01 to 500mg/kg weight in patients/sky, and it can give with single or multiple dosage.Preferably, dosage level will be about 0.1 to about 250mg/kg/ day; More preferably from about 0.5 to about 100mg/kg/ day.The appropriate dosage level can be about 0.01 to 250mg/kg/ day, about 0.05 to 100mg/kg/ day or about 0.1 to 50mg/kg/ day.In this scope, dosage can be 0.05 to 0.5,0.5 to 5 or 5 to 50mg/kg/ days.For oral administration, preferably the form with tablet provides composition, wherein tablet comprises 1.0 to 1000 milligrams active ingredient, especially 1.0,5.0,10.0,15.0,20.0,25.0,50.0,75.0,100.0,150.0,200.0,250.0,300.0,400.0,500.0,600.0,750.0,800.0,900.0 and 1000.0 milligrams active ingredient is used for patient's to be treated dosage is carried out the symptom adjustment.Can be based on 1 to 4 time/day, preferred once or twice/day scheme give this compound.
Yet, be to be understood that, be used for the concrete dosage level of any particular patient and dose frequency and can change and will depend on various factors, comprise the metabolic stability of the activity of the particular compound that is adopted, described compound and time span, age, body weight, general health state, sex, diet, the mode that gives and time, excretion rate, drug regimen, the seriousness of particular disorder and the host who stands to treat of effect.
In order more to be expressly understood characteristic of the present invention, preferred form of the present invention is described now with reference to following non-limiting example.
Embodiment
Method
The HPLC condition
Carrying out all HPLC with Waters 2690 Alliance System measures.
Method 1
Post:
Waters Exterra C18 post (Part#186000410), under 30 ℃, flow velocity 0.4mL/ minute, in 254nM place measure spectrum
Buffer reagent:
Buffer reagent A:100% water, buffer reagent B:100% acetonitrile, the moisture TFA of buffer reagent C:2%
Gradient: (linear gradient curve 6)
Figure G2007800515627D00171
Method 2
Post:
Merck C18Chromolith post (Part#1.02129.0001), under 30 ℃, flow velocity 4mL/ minute, in 254nM place measure spectrum
Buffer reagent:
Buffer reagent A:100% water, buffer reagent B:100% acetonitrile, the moisture TFA of buffer reagent C:2%
Gradient: (linear gradient curve 6)
Figure G2007800515627D00172
Method 3
Post:
Merck C18 Chromolith post (Part#1.02129.0001), under 30 ℃, flow velocity 4mL/ minute, in 254nM place measure spectrum
Buffer reagent:
Buffer reagent A:100% water, buffer reagent B:100% acetonitrile, the moisture TFA of buffer reagent C:2%
Gradient: (linear gradient curve 6)
Figure G2007800515627D00173
Method 4
Post:
Merck C18Chromolith post (Part#1.02129.0001), under 30 ℃, flow velocity 4mL/ minute, in 254nM place measure spectrum
Buffer reagent:
Buffer reagent A:100% water, buffer reagent B:100% acetonitrile, the moisture TFA of buffer reagent C:2%
Gradient: (linear gradient curve 6)
Figure G2007800515627D00181
Method 5
Post:
Phenomenex Gemini C18 post (Part#344382-3), under 30 ℃, flow velocity 0.4mL/ minute, in 254nM place measure spectrum
Buffer reagent:
Buffer reagent A:100% water, buffer reagent B:100% acetonitrile, the moisture TFA of buffer reagent C:2%
Gradient: (linear gradient curve 6)
Figure G2007800515627D00182
Method 6
Post:
Phenomenex Gemini C18 post (Part#344382-3), under 30 ℃, flow velocity 0.4mL/ minute, in 254nM place measure spectrum
Buffer reagent:
Buffer reagent A:100% water, buffer reagent B:100% acetonitrile, the moisture TFA of buffer reagent C:2%
Cotton degree: (linear gradient curve 6)
Figure G2007800515627D00191
Method 7
Post:
Waters
Figure G2007800515627D00192
C18 post (Part No WAT045905), under 25 ℃, flow velocity 1mL/ minute, in 254nM place measure spectrum
Buffer reagent:
Buffer reagent A:100% acetonitrile, the moisture TFA of buffer reagent B:0.1%
Gradient: (linear gradient curve 6)
Figure G2007800515627D00193
General diagram 1: synthetic
General procedure 1:Organic Preparations and Procedures International, 22 (4), 1990, the reorganization of 532-534
As in diagram 1, aminocompound can react with the suitable analogue of fumaric acid derivatives or fumaric acid, and wherein for example Acetyl Groups can be by other suitable leavings groups such as tosyl group or methylsulfonyl replacement (replacement).Reaction can be carried out in The suitable solvent such as methyl alcohol, DME, DMA, DMSO, chloroform, THF Huo diox.Reaction can be heated or carry out microwave radiation (referring to for example B.R.Roberts﹠amp; C.R.Strauss, Acc.Chem.Res.2005,38,653-661, " Toward Rapid, ' Green ' Predictable Microwave-assisted Synthesis ").Reaction can or under the situation that has the acid of catalytic amount or alkali to exist not carried out.
General diagram 2: interchangeable synthetic 1
Figure G2007800515627D00201
General diagram 3: interchangeable Synthetic 2
Figure G2007800515627D00211
Further reaction: diagram 4
Figure G2007800515627D00212
By the known method of those of ordinary skills as implied above, can further making wherein, substituting group is the compound reaction of halogen, wherein ' and the palladium coupling ' comprise that reaction is as Suzuki, Buchwald, Heck or Sonogashira and nucleophilic aromatic replacement (referring to, for example, at L.S.Hegedus, " Transition Metals in the Synthesis ofComplex Organic Molecules ", University Science books, 1994, first version or M.Smith " Organic synthesis ", 2001, McGraw-Hill Science, the reaction of describing in the second edition), its to use the suitable form of metal catalyst such as palladium and reagent 1 alternatively be R 1The derivative (R that for example comprises halogen or boride 1Derivative).
Diagram 5
Figure G2007800515627D00221
Can protect OH by method known to those skilled in the art; shown in diagram (5); for example wherein ' P ' can be that benzyl is (referring to I.Stansfield et al; ' Active siteinhibitors of HCV NS5B polymerase ' Bio-Org.Med-Chem.Lett; 2004; 14; 5085-5088) or suitable blocking group; as at " Protective Groups inOrganic Synthesis " Theodora Greene and Peter Wuts; third edition; Wiley Interscience, described in 1999.
Diagram 6
Figure G2007800515627D00222
Embodiment 1: the preparation of diacetoxyl dimethyl fumarate
Figure G2007800515627D00223
According at OPPI, 22 (4), 1990, the program (step) described in the 532-534.
(10.7g, the turbid solution a little of stirring 72.5mmol) cools off (ice/water-bath) to the Dihydroxyfumaric acid in anhydrous methanol (50mL) under nitrogen.By means of syringe, through 20 minutes, under the surface of methanol solution, add thionyl chloride (10.5mL, 144mmol).After adding, remove cooling bath and also at room temperature stirred the mixture 3 days.By filtering the resulting throw out of collection and washing with cold methanol (10mL), water (80mL).Obtain the Dihydroxyfumaric acid dimethyl ester, as white solid (11.8g).Merge this material (10.8g) and methylvinyl acetate (36mL) and under nitrogen, when stirring, be heated to and refluxed 8 hours.Reaction is cooled to room temperature and is kept under 0 ℃ spend the night.By filtering the resulting throw out of collection and washing so that the diacetoxyl dimethyl fumarate to be provided, as white solid (6.4g) with cold methanol (5mL).
The preparation of embodiment 2:7-bromo-3-hydroxyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylate methyl ester
Figure G2007800515627D00231
Be incorporated in 2-amino-5-bromo-pyridine in the dried methyl alcohol (10mL) (664mg, 3.85mmol), the diacetoxyl dimethyl fumarate (1.00g, 3.84mmol) and Glacial acetic acid (10) and be heated to backflow.After 48 hours, reaction is cooled to room temperature and under vacuum, concentrates.(2mL) joins in the residue with ethyl acetate, it was by sonication 2 minutes, collect resulting throw out and use cold ethyl acetate (1mL) washing by filtering, then with the pump drying so that 7-bromo-3-hydroxyl-4-oxo-4H-pyrido [1 to be provided, 2-α] pyrimidine-2-carboxylate methyl ester, as yellow solid (24mg, 2%): 1H NMR (300MHz, CDCl 3) δ 4.19 (3H, s, OCH 3), 7.53 (2H, d, J=1.4Hz, H8 and H9), 9.00 (1H, d, J=1.4Hz, H6), 10.53 (1H, br s, OH).
MS(ESI +)m/z?299(M[Br 79]+1)，301(M[Br 81]+1)，
HPLC Method 196.0%/4.30 minute.
The preparation of embodiment 2.1:3-hydroxyl-7-methyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylate methyl ester
1H?NMR(300MHz，CDCl 3)δ2.40(3H，s，CH 3)，4.11(3H，s，OCH 3)，7.39(1H，dd，J＝9.1Hz，1.8Hz，H8)，7.60(1H，d，J＝9.1Hz，H9)，8.68(1H，m，H6)。
MS(ESI +)m/z235(M+1)，(ESI -)m/z233(M-1)
HPLC Method 1100%/3.74 minute.
The preparation of embodiment 2.2:7-chloro-3-hydroxyl-4-oxo-4H-pyrido [l, 2-a] pyrimidine-2-carboxylate methyl ester
Figure G2007800515627D00242
1H?NMR(300MHz，CDCl 3)δ8.74(1H，dd，J＝2.4Hz，09Hz，H7)，7.52(1H，d，J＝9.4Hz，H9)，7.41(1H，dd，J＝9.4Hz，2.4Hz，H8)，3.98(3H，s，CH 3)。
MS(ESI +)m/z255(M+1)，(ESI -)m/z253(M-1)
HPLC Method 196%/4.14 minute.
The preparation of embodiment 2.3:3-hydroxyl-7-morpholine-4-base-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylate methyl ester
Figure G2007800515627D00251
By adopting at J.Med.Chem., 2005,48 (7), the program of describing among the 2388-2406 prepares 5-morpholine-4-base-pyridine-2-base amine.Briefly, under 60-70 ℃, make morpholine among 5-bromo-2-nitro-pyridine and the DMSO and salt of wormwood react 4-(6-nitro-pyridin-3-yl)-morpholine so that 84% productive rate to be provided.Carbon with the palladium load under nitrogen atmosphere reduces with the basic amine of 5-morpholine-4-base-pyridine-2-that 70% productive rate is provided.By adopting the program of describing in embodiment 2 to convert it into 3-hydroxyl-7-morpholine-4-base-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylate methyl ester of 25% productive rate, wherein Glacial acetic acid replaces p-methyl benzenesulfonic acid.
1H?NMR(300MHz，D 6-DMSO)：δ3.22(4H，t，J＝5.0Hz，-NCH 2CH 2O-)，3.89(4H，t，J＝5.0Hz，-NCH 2CH 2O-)，4.10(3H，s，OCH 3)，7.45(1H，dd，J＝9.9，2.7Hz，H8)，7.63(1H，d，J＝9.9Hz，H9)，8.17(1H，d，J＝2.7Hz，H6)，10.32(1H，s，OH)
MS(ESI +)m/z305(M+1)
HPLC Method 797.4%/11.7 minute.
The preparation of embodiment 2.4:3-hydroxyl-8-morpholine-4-base-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylate methyl ester
Figure G2007800515627D00252
2-pyridine carboxylic acid and thionyl chloride and methyl alcohol are reacted 4-chloro-2-pyridine carboxylic acid methyl esters (it is hydrolyzed) and hydrochloride to be provided and to stand Ku Ertisi and reset.Dissociating of Boc blocking group provides 2-amino-4-chloropyridine.The program of describing in W02006040520 is used in the position 4 introducing morpholines, adopts the program of describing in embodiment 2, and ester in the middle of it is cyclized into wherein reacts under 60 ℃.
1H?NMR(300MHz，DMSO-d 6)δ3.43(t，J＝4.7Hz，4H)，3.73(t，J＝4.7Hz，4H)，3.85(s，3H)，6.67(d，J＝2.4Hz，1H)，7.27(dd，J＝8.2Hz，2.5Hz，1H)，8.59(d，J＝8.2Hz，1H)，9.29-9.63(brs，1H)
MS(ESI +)m/z328(M+23)
The preparation of embodiment 2.5:3-hydroxyl-9-morpholine-4-base-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylate methyl ester
Figure G2007800515627D00261
The program that to describe in embodiment 2.3 is used for 3-bromo-2-nitro-pyridine so that 2-amino-3-morpholino pyridine to be provided, and is converted to the ester of expectation by adopt the program of describing in embodiment 2.
1H?NMR(300MHz，CDCl 3)δ3.45-3.56(m，4H)，4.00-4.11(m，7H)，6.97(t，J＝7.3Hz，1H)，7.22(d，J＝7.3Hz，1H)，8.63(dd，J＝7.2，1.1Hz，1H)，10.31(s，1H)
MS(ESI +)m/z328(M+23)
The preparation of embodiment 2.6:3-hydroxyl-4-oxo-7-piperidines-l-base-4H-pyrido [l, 2-α] pyrimidine-2-carboxylate methyl ester
Figure G2007800515627D00262
Except using piperidines, adopted the program of in embodiment 2.3, describing.By adopting the program of in embodiment 2, describing resulting 2-amino-5-piperidines and pyridine to be changed into the ester of expectation.Except adopting trimethyl-acetyl chloride (pivaloyl chloride), (step 1) makes the crude product reaction, so that the product of expectation to be provided by the program of employing in embodiment 17.1.
1H?NMR(300MHz，CDCl 3)δ1.43(s，9H)，1.58-1.82(m，6H)，3.24(t，J＝4.8Hz，4H)，3.96(s，3H)，7.64-7.78(m，2H)，8.30(d，J＝1.8Hz，1H)
Embodiment 3:3-cyano group-6-hydroxyl-7-oxo-l, the preparation of 7-dihydro-pyrazolo [l, 5-α] pyrimidine-5-carboxylic acid's methyl esters
Figure G2007800515627D00271
With 3-amino-4-pyrazoles nitrile (400mg, 3.7mmol) and Dihydroxyfumaric acid (978mg 5.5mmol) is dissolved in the Glacial acetic acid (5mL) and with mixture heating up to 100 ℃.After two days, analyze by HPLC and to finish reaction, and be cooled to room temperature.Add ethyl acetate (10mL) to cause the product precipitation and to collect resulting throw out by filtering.Separated product, productive rate are 62% (534mg).
1H NMR (300MHz, D 6-DMSO): δ 3.90 (3H, s, OCH 3) and 8.39 (1H, s, CHC[CN]).
MS(ESI +)m/z?235(M+1)
HPLC Method 289.3%/1.07 minute.
Embodiment 3.1: prepare 3-cyano group-6-hydroxyl-7-oxo-l with the 4-flunamine, 7-dihydro-pyrazolo [l, 5-α] pyrimidine-5-carboxylic acid 4-fluoro-benzyl amide compound
Figure G2007800515627D00281
With 3-cyano group-6-hydroxyl-7-oxo-1,7-dihydro-pyrazolo [1,5-a] pyrimidine-5-carboxylic acid's methyl esters (100mg, 0.427mmol) be suspended among the MeOH (10mL) and to wherein adding the fluorine-based benzylamine of 4-(122 μ L, 1.07mmol), and under refluxing reacting by heating two days, be cooled to room temperature then and filter.Concentrated filtrate and recrystallization self-heating methyl alcohol product (25mg, 18%) so that yellow solid to be provided.
1H?NMR(300MHz，D6-DMSO)：δ4.01(2H，s，H 3N +CH 2Ph)，4.48(2H，d，J＝6.6Hz，[C＝O]NHCH 2)，7.14-7.49(8H，m，Ar-CH)，8.08(1H，s，CHC[CN])，9.13(1H，t，J＝6.6Hz，[C＝O]NHCH 2)。
MS(ESI +)m/z?328[M+H] +
Embodiment 3.2: with 3,4-two chloro-benzylamines prepare 3-cyano group-6-hydroxyl-7-oxo-l, 7-dihydro-pyrazolo [l, 5-α] pyrimidine-5-carboxylic acid 3,4-two chloro-benzyl amide compounds
By adopting the program in embodiment 3.1 to prepare compound
1H?NMR(300MHz，D6-DMSO)：δ8.45(1H，s，Ar-CH)，7.79-7.57(4H，m，Ar-CH)，7.39-7.32(2H，m，Ar-CH)，4.78(2H，s，H 3N +CH 2)，4.49(2H，d，J＝6.3Hz，NHCH 2)。
MS(ESI +)m/z379[M+H] +
Embodiment 3.3:6-hydroxyl-7-oxo-2-phenyl-l, the preparation of 7-dihydro-pyrazolo [l, 5-a] pyrimidine-5-carboxylic acid's methyl esters
By adopting the program in embodiment 3 to prepare compound.
1H?NMR(300MHz，D6-DMSO)：δ3.94(3H，s，OCH 3)，6.47(1H，s，CHC[Ph])，7.42-7.51(3H，m，Ar-CH)，7.95(2H，d，J＝6.6Hz，Ar-CH)。
MS(ESI +)m/z286[M+1] +。
HPLC Method 295.6%/1.24 minute.
Embodiment 3.4: prepare 6-hydroxyl-7-oxo-2-phenyl-l with 4-fluoro-benzylamine, 7-dihydro-pyrazolo [l, 5-α] pyrimidine-5-carboxylic acid 4-fluoro-benzyl amide compound
Figure G2007800515627D00292
By adopting the program in embodiment 3.1 to prepare compound.
1H?NMR(300MHz，D6-DMSO)：δ4.01(2H，s，NH 3CH 2Ph)，4.47(2H，d，J＝6.0Hz，[C＝O]NHCH 2)，6.03(1H，s，CHC[Ph])，7.11-7.51(11H，m，Ar-CH)，7.91(2H，d，J＝7.2Hz，Ar-CH)。
MS(ESI +)m/z379[M+H] +
Embodiment 3.5:6-hydroxyl-7-oxo-2-phenyl-l, 7-dihydro-pyrazolo [l, 5-α] pyrimidine-5-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00301
By adopting the program in embodiment 3.1 to prepare compound.
1H?NMR(300MHz，D6-DMSO)：δ4.58(2H，d，J＝6.0Hz，NCH 2)，6.49(1H，s，CHC[Ph])，7.29-7.67(6H，m，Ar-CH)，7.92(2H，d，J＝7.2Hz，Ar-CH)，9.20(1H，t，J＝6.0Hz，[C＝O]NHCH 2)。
MS(ESI +)m/z429[M+H] +
Embodiment 3.6:6-hydroxyl-7-oxo-l, the preparation of 7-dihydro-pyrazolo [l, 5-a] pyrimidine-5-carboxylic acid's methyl esters
Figure G2007800515627D00302
By adopting the program in embodiment 3 to prepare compound.
1H?NMR(300MHz，D 6-DMSO)：δ7.96(1H，s，CHCHNH)，6.05(1H，d，J＝1.8Hz，CHCHNH)，3.87(3H，s，OCH 3)。
MS(ESI +)m/z210(M+1)
HPLC Method 397%/1.18 minute
Embodiment 3.7:6-hydroxyl-7-oxo-l, the preparation of 7-dihydro-pyrazolo [l, 5-α] pyrimidine-5-carboxylic acid 4-chloro-benzyl acid amides
Figure G2007800515627D00311
By adopting the program in embodiment 3.1 to prepare compound.
1H?NMR(300MHz，D 6-DMSO)：δ10.0(1H，s，OH)，9.35(1H，t，J＝6.3Hz，NHCH 2)，7.93(1H，d，J＝8.4Hz，Ar-CH)，7.68(1H，d，J＝8.4Hz，Ar-CH)，7.38-7.26(4H，m，Ar-CH)，4.30(2H，d，J＝6.3Hz，NHCH 2)。
MS (ESI +) m/z do not have ionization
Embodiment 3.8:l-(4-fluoro-benzyl)-6-hydroxyl-7-oxo-l, the preparation of 7-dihydro-pyrazolo [l, 5-α] pyrimidine-5-carboxylic acid's methyl esters
Figure G2007800515627D00312
By adopting the program in embodiment 3 to prepare compound.
1H?NMR(300MHz，D 6-DMSO)：δ8.48(1H，d，J＝3.6Hz，Ar-CH)，7.23-7.08(4H，m，Ar-CH)，6.47(1H，d，J＝3.6Hz，Ar-CH)，5.76(2H，s，CH 2Ar)，3.81(3H，s，OCH 3)。
MS(ESI +)m/z318[M+H] +
Embodiment 3.9:l-(4-fluoro-benzyl)-6-hydroxyl-7-oxo-l, the preparation of 7-dihydro-pyrazolo [l, 5-α] pyrimidine-5-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D00321
By adopting the program in embodiment 3.1 to prepare compound.
1H?NMR(300MHz，D 6-DMSO)：δ11.96(1H，s，OH)，9.53(1H，s，NH)，8.48(1H，d，J＝3.3Hz，Ar-CH)，7.36(1H，d，J＝5.7Hz，Ar-CH)，7.33(1H，d，J＝5.4Hz，Ar-CH)，7.23(1H，d，J＝5.4Hz，Ar-CH)，7.20(1H，d，J＝5.4Hz，Ar-CH)，7.18-7.0(4H，m，Ar-CH)，6.40(2H，d，J＝3.6Hz，CH 2NH)，5.73(2H，s，CH 2Ar)。
MS(ESI +)m/z411[M+H] +。
Embodiment 3.10:l-(4-fluoro-benzyl)-6-hydroxyl-7-oxo-l, 7-dihydro-pyrazolo [l, 5-α] pyrimidine-5-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00322
By adopting the program in embodiment 3.1 to prepare compound.
1H NMR (300MHz, D 6-DMSO): δ 11.83 (1H, s, OH), 9.57 (1H, t, J=6.9Hz, NHCH 2), 8.50 (1H, d, J=3.6Hz, Ar-CH), 7.56 (1H, m, Ar-CH), 7.31-7.04 (5H, m, Ar-CH), 6.41 (1H, d, J=3.6Hz, Ar-CH), 5.74 (3H, m, CH 2Ar and Ar-CH), 4.41 (2H, d, J=6.9Hz, CH 2NH).
MS(ESI +)m/z?461[M] +。
Embodiment 3.11:6-hydroxyl-l-methyl-7-oxo-l, the preparation of 7-dihydro-pyrazolo [l, 5-α] pyrimidine-5-carboxylic acid's methyl esters
Figure G2007800515627D00331
By adopting the program in embodiment 3 to prepare compound, and use it for next step and need not to be further purified.
Embodiment 3.12:6-hydroxyl-l-methyl-7-oxo-l, 7-dihydro-pyrazolo [l, 5-α] pyrimidine-5-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00332
By adopting the program in embodiment 3.1 to prepare compound.
1H?NMR(300MHz，D 6-DMSO)：δ9.77(1H，s，NH)，8.12(1H，d，J＝3.3Hz，Ar-CH)，7.60-7.52(2H，m，Ar-CH)，7.34-7.30(1H，m，Ar-CH)，6.32(1H，d，J＝3.6Hz，Ar-CH)，4.46(2H，d，J＝5.7Hz，CH 2NH)，4.02(3H，s，CH 3)。
MS (ESI +) m/z 367 and 369[M+H] +
Embodiment 3.13:6-hydroxyl-l-methyl-7-oxo-l, the preparation of 7-dihydro-pyrazolo [l, 5-α] pyrimidine-5-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D00333
By adopting the program in embodiment 3.1 to prepare compound.
1H?NMR(300MHz，D 6-DMSO)：δ11.8(1H，s，OH)，9.54(1H，s，NH)，8.12(1H，d，J＝3.3Hz，Ar-CH)，7.38(1H，d，J＝9.3Hz，Ar-CH)，7.35(1H，d，J＝8.7Hz，Ar-CH)，7.15(1H，d，J＝9.3Hz，Ar-CH)，7.12(1H，d，J＝8.7Hz，Ar-CH)，6.33(1H，d，J＝3.6Hz，Ar-CH)，4.45(2H，d，J＝6.6Hz，CH 2NH)，4.05(3H，s，CH 3)。
MS(ESI +)m/z317[M+H] +。
Embodiment 3.14:3-bromo-6-hydroxyl-7-oxo-l, the preparation of 7-dihydro-pyrazolo [l, 5-α] pyrimidine-5-carboxylic acid's methyl esters
Figure G2007800515627D00341
By adopting the program in embodiment 3 to prepare compound, and use it for next step and need not to be further purified.
Embodiment 3.15:3-bromo-6-hydroxyl-7-oxo-l, 7-dihydro-pyrazolo [1,5-α] pyrimidine-5-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00342
By adopting the program in embodiment 3.1 to prepare compound.
1H?NMR(300MHz，D 6-DMSO)：δ4.78(2H，s，CH 2NH)，7.35(1H，d，J＝8.1Hz，CHCHC[Cl])，7.60(1H，d，J＝8.1Hz，CHCHC[Cl])，7.61(1H，d，J＝1.8Hz，NHCHC[Br])，7.75(1H，s，[C]CHC[Cl])，8.00(1H，s，CH 2NH)，8.50(1H，s，OH)。
MS(ESI +)m/z473[M+MeCN] +。
Embodiment 3.16:3-bromo-6-hydroxyl-7-oxo-l, the preparation of 7-dihydro-pyrazolo [1,5-α] pyrimidine-5-carboxylic acid 2-fluoro-benzyl acid amides
Figure G2007800515627D00351
By adopting the program in embodiment 3.1 to prepare compound.
1H NMR (300MHz, D 6-DMSO): δ 4.89 (2H, s, CH 2NH), 7.21-7.45 (5H, m, 4 * Ar-CH and 1 * NH), 8.04 (1H, dd, J=8.1,7.4Hz, Ar-CH), 8.74 (1H, s, CH 2NH).
MS (ESI +) m/z do not have ionization.
Embodiment 3.17:6-hydroxyl-7-oxo-3-phenyl-l, the preparation of 7-dihydro-pyrazolo [1,5-α] pyrimidine-5-carboxylic acid's methyl esters
Figure G2007800515627D00352
Be cooled to chamber Gentle ethanol: before Virahol (surpassing) sonication 10 minutes, 100 ℃ down heating phenylpyrazole amine (100mg, 0.63mmol), the diacetoxyl fumarate (180mg, 0.69mmol) and tosic acid (5mg) 20 minutes.Collect resulting throw out and use washing with alcohol.Separated product is as yellow solid (47mg, 26%).
1H?NMR(300MHz，D 6-DMSO)：δ8.53(1H，s，Ar-CH)，7.82(2H，d，J＝6.9Hz，Ar-CH)，7.47(2H，m，Ar-CH)，7.29(1H，m，Ar-CH)，3.89(3H，s，OCH 3)。
MS(ESI +)m/z286[M+H] +。
Embodiment 3.18:6-hydroxyl-7-oxo-3-phenyl-l, 7-dihydro-pyrazolo [1,5-α] pyrimidine-5-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00361
By adopting the program in embodiment 3.1 to prepare compound.
1H?NMR(300MHz，D 6-DMSO)：δ9.29(1H，m，NH)，8.48(1H，bs，CHNH)，7.63(1H，d，J＝1.8Hz，Ar-CH)，7.61(2H，m，Ar-CH)，7.59(1H，s，Ar-CH)，7.47-7.35(4H，m，Ar-CH)，7.24(1H，m，Ar-CH)，4.59(2H，d，J＝6.3Hz，NHCH 2)。
MS(ESI +)m/z?429[M] +。
Embodiment 3.19:3-cyano group-l-(4-fluoro-benzyl)-6-hydroxyl-7-oxo-l, the preparation of 7-dihydro-pyrazolo [1,5-α] pyrimidine-5-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D00362
Under nitrogen and room temperature, with portion with sodium hydride (11.0mg, 0.367mmol) join the 3-cyano group-6-hydroxyl-7-oxo-1 in DMF (2mL), 7-dihydro-pyrazolo [1,5-a] (100mg is in stirred suspension 0.306mmol) for pyrimidine-5-carboxylic acid 4-fluoro-benzyl amide compound and the fluorine-based benzylamine of 4-(embodiment 3.1).(40 μ L 0.366mmol) and before 90 ℃ of following heated mixt two days, stirred the mixture 30 minutes to fluorobenzyl chloride adding.Thereafter, reaction is cooled to room temperature and at ethyl acetate (10mL) and aqueous hydrochloric acid (1M distributes between 10mL).Separate organic layer and use ethyl acetate (3 * 10mL) extraction water layers.Water (3 * 10mL), the organism that merges of salt solution (10mL) washing, concentrate then.By column chromatography (95: 5: 1 methylene dichloride: methyl alcohol: ammoniacal liquor) come the product (50mg, 36%) of purifying residue so that expectation to be provided.
1H?NMR(300MHz，D 6-DMSO)：δ8.11(1H，s，CHC[CN])，7.45(1H，d，J＝6.0Hz，Ar-H)，7.42(1H，d，J＝6.0Hz，Ar-H)，7.36(1H，d，J＝5.4Hz，Ar-H)，7.34(1H，d，J＝5.4Hz，Ar-H)，7.15(1H，d，J＝9.3Hz，Ar-H)，7.09(1H，d，J＝8.7Hz，Ar-H)，7.05(1H，d，9.3Hz，Ar-H)，7.03(1H，d，J＝8.7Hz，Ar-H)，4.96(2H，s，CH 2N)，4.38(2H，d，J＝6.3Hz，NHCH 2)。
MS(ESI +)m/z434[M+H] +。
Embodiment 4:6-hydroxy-2-methyl-7-oxo-7H-isoxazole is the preparation of [2,3-α] pyrimidine-5-carboxylic acid's methyl esters also
Figure G2007800515627D00371
In the bottle of adding a cover, merge 5-methyl-isoxazole-3-base amine (392mg, 3.99mmol), (1.04g 3.99mmol) and tosic acid (10mg), and is heated to 100 ℃ to the diacetoxyl dimethyl fumarate.After 5 hours, reaction is cooled to room temperature and ethanol (2.5mL) and isopropyl ether (2.5mL) are joined in the residue, it is carried out (surpassing) sonication 15 minutes.Collect resulting throw out and, use the pump drying then by filtering with cold ethanol (5mL) washing, so that also [2,3-α] pyrimidine-5-carboxylic acid's methyl esters (331mg, 37%) of 6-hydroxy-2-methyl-7-oxo-7H-isoxazole to be provided: 1H NMR (300MHz, D 6DMSO) δ 2.50 (3H, s, CH 3), 3.84 (3H, s, OCH 3), 6.67 (1H, s, H3), 10.31 (1H, br s, OH).
MS(ESI +)m/z225(M+1)
HPLC Method 499.3%/0.52 minute.
Embodiment 5:3-hydroxyl-4-oxo-4, the preparation of 10-dihydro-benzo [4,5] imidazo [l, 2 α] pyrimidine-2-carboxylate methyl ester
Figure G2007800515627D00372
In sealed tube, 100 ℃ down the amino benzoglyoxalines of heating 2-(200mg, 1.50mmol), dimethoxy diacetoxyl fumarate (430mg, 1.65mmol) and tosic acid (5mg) 2 hours.Also (surpass) sonication 10 minutes with ethanol/isopropyl ether (5mL) development residue.Collect resulting throw out and recrystallization self-heating acetonitrile to provide 3-hydroxyl-4-oxo-4,10-dihydro-benzo [4,5] imidazo [1,2 α] pyrimidine-2-carboxylate methyl ester (153mg, 39%).
1H NMR (300MHz, D 6-DMSO): δ 3.90 (3H, s, OCH 3), 7.10 (1H, m, Ar-CH), 7.25-7.51 (3H, m, 2 * Ar-CH and NH) and 8.43 (1H, d, J=7.8Hz, Ar-CH).
MS(ESI +)m/z?260(M+1)
The preparation of embodiment 6:7-bromo-3-hydroxyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D00381
Under agitation, will (20mg, 0.07mmol) and to fluorine-based benzylamine (19 μ L 0.17mmol) be heated to backflow at 7-bromo-3-hydroxyl-4-oxo-4H-pyrido [1, the 2-α] pyrimidine-2-carboxylate methyl ester in the dried methyl alcohol (4mL).Make progress by the HPLC monitoring reaction.After 6 hours, reaction is cooled to room temperature and under vacuum, concentrates.With ether (2mL) development residue and by filtering the collecting precipitation thing, use ether (10mL) washing then and use the pump drying, to provide as yellowish brown solid 7-bromo-3-hydroxyl-4-oxo-4H-pyrido [1,2-a] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides (24mg, 92%).
1H?NMR(300MHz，D 6DMSO)δ4.51(2H，s，NCH 2)，7.33(6H)，8.57(1H，m，H6)，11.21(1H，br?s，NH)。
MS(ESI +)m/z?392(M[Br 79]+1)，394(M[Br 81]+1)，
HPLC Method 199.6%/6.5 minute.
By adopting the program of in embodiment 6, describing, obtain following compound (6.1-6.13):
The preparation of embodiment 6.1:3-hydroxyl-7-methyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
MS(ESI +)m/z?328(M+1)，
HPLC Method 194.8%/6.20 minute.
Embodiment 6.2:3-hydroxyl-7-methyl-4-oxo-4H-pyrido [l, 2-a] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00392
MS(ESI +)m/z?378(M[Cl 35]+1)。
HPLC Method 1100%/6.74 minute.
The preparation of embodiment 6.3:7-chloro-3-hydroxyl-4-oxo-4H-pyrido [l, 2-a] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D00393
1H?NMR(300MHz，CDCl 3)δ9.95(1H，bs，NH a)，8.931H，app.t，NH b)，8.62(1H，s，H6 a)，7.58(1H，d，J＝.4Hz，H8)，7.45-7.27(2H，m，ArH)，7.26-7.21(2H，m，ArH)，7.07-6.99(2H，m，ArH)，6.99-6.90(2H，m，ArH)，4.52(0.32H，d，＝.4Hz，CH 2a)，4.31(0.68H，d，J?6.6Hz，CH 2b)。
MS(ESI +)m/z348(M+1)，(ESI -)m/z346(M-1)
HPLC Method 193%/6.35 minute.
Embodiment 6.4:7-chloro-3-hydroxyl-4-oxo-4H-pyrido [l, 2-a] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
1H?NMR(300MHz，CDCl 3)δ9.95(1H，bs，NH a)，9.13(1H，app.t，NH b)，8.65(1H，s，H6 a)，7.57(1H，d，J?8.4Hz，H8)，7.40-7.33(2H，m，ArH)，7.26-7.24(1H，m，ArH)，7.153(1H，dd，J＝8.1Hz，1.8Hz，ArH)，7.05(1H，d，J＝7.8Hz，ArH)，4.51(0.41H，d，J＝6.3Hz，CH 2a)，4.30(0.59H，d，J＝6.3Hz，CH 2b)
MS(ESI +)m/z400(M+1)，(ESI -)m/z396(M-1)
HPLC Method 191%/6.89 minute.
Embodiment 6.5:3-hydroxyl-4-oxo-4, the preparation of 10-dihydro-benzo [4,5] imidazo [l, 2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D00402
1H NMR (300MHz, D 6-DMSO): δ 4.51 (2H, d, J=6.3Hz, NHCH 2), 7.14-7.50 (7H, m, Ar-CH), 8.44 (1H, d, J=8.1Hz, Ar-CH) and 9.27 (1H, t, J=6.3Hz, NHCH 2).
MS(ESI +)m/z353(M+1)
HPLC Method 592%/3.10 minute.
Embodiment 6.6:6-hydroxy-2-methyl-7-oxo-7H-isoxazole is the preparation of [2,3-α] pyrimidine-5-carboxylic acid 4-fluoro-benzyl acid amides also
Figure G2007800515627D00411
1H?NMR(300MHz，D 6DMSO)δ2.49(3H，s，CH 3)，4.48(2H，d，J＝5.9Hz，NCH 2)，6.50(s，1H，H3)，6.95(2H，m，ArH)，7.30(2H，m，ArH)，9.14(1H，br?s，NH)。
MS(ESI +)m/z318(M+1)，(ESI -)m/z316(M-1)
HPLC Method 190%/5.50 minute.
Embodiment 6.7:6-hydroxy-2-methyl-7-oxo-7H-isoxazole is [2,3-α] pyrimidine-5-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl acid amides
1H?NMR(300MHz，D 6DMSO)δ2.49(3H，s，CH 3)，4.48(2H，d，J＝5.9Hz，NCH 2)，7.30(1H，dd，J＝8.2，2.3Hz，ArH)，7.52(2H，m，ArH)，10.45(1H，br?s，NH)。
MS(ESI +)m/z368(M[Cl 35，Cl 35]+1)，(ESI -)m/z366(M[Cl 35，Cl 35]-1)
HPLC Method 492%/1.83 minute.
The preparation of embodiment 6.8:3-hydroxyl-7-morpholine-4-base-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D00413
1H NMR (300MHz, D6-DMSO): δ 12.13 (1H, s, OH), 9.67 (1H, t, J=6.9Hz, NHCH 2), 8.00 (1H, s, CHC[morpholine]), 7.85 (1H, d, J=9.6Hz, CHCHC[morpholine]), 7.50 (1H, d, J=9.6Hz, CHCHC[morpholine]), 7.39 (2H, m, Ar-CH), 7.16 (2H, m, Ar-CH), 4.50 (2H, d, J=6.9Hz, NHCH 2), 3.76 (4H, m, CH 2OCH 2) and 3.16 (4H, m, CH 2NCH 2).
MS(ESI +)m/z397(M+1)
HPLC Method 698%/6.40 minute
The preparation of embodiment 6.9:3-hydroxyl-7-morpholine-4-base-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 4-methoxyl group-benzyl acid amides
Figure G2007800515627D00421
1H NMR (300MHz, D6-DMSO): δ 12.25 (1H, s, OH), 9.55 (1H, t, J=6.9Hz, CH 2NH), 7.99 (1H, s, CHC[morpholine]), 7.84 (1H, d, J=9.6Hz, CHCHC[morpholine]), 7.50 (1H, d, J=9.6Hz, CHCHC[morpholine]), 7.29 (2H, d, J=8.4Hz, Ar-CH), 6.91 (2H, d, J=8.4Hz, Ar-CH), 4.44 (2H, d, J=6.9Hz, CH 2NH), 3.79-3.72 (4H, m, CH 2OCH 2), 3.70 (3H, s, OCH 3) and 3.18 (4H, m, CH 2NCH 2).
MS(ESI +)m/z411(M+1)
HPLC Method 699%/6.21 minute
The preparation of embodiment 6.10:3-hydroxyl-7-morpholine-4-base-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid benzyl acid amides
Figure G2007800515627D00422
1H NMR (300MHz, D6-DMSO): δ 12.17 (1H, s, OH), 9.66 (1H, t, J=6.3Hz, CH 2NH), 7.99 (1H, s, CHC[morpholine]), 7.85 (1H, d, J=9.6Hz, CHCHC[morpholine]), 7.50 (1H, d, 9.6Hz, CHCHC[morpholine]), and 7.38-7.25 (5H, m, Ar-CH), 4.52 (2H, d, J=6.3Hz, CH 2NH), 3.77 (4H, m, CH 2OCH 2) and 3.18 (4H, m, CH 2NCH 2).
MS(ESI +)m/z381(M+1)
HPLC Method 697%/6.32 minute
The preparation of embodiment 6.11:3-hydroxyl-7-morpholine-4-base-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 4-chloro-benzyl acid amides
Figure G2007800515627D00431
1H NMR (300MHz, D6-DMSO): δ 12.10 (1H, s, OH), 9.69 (1H, t, J=6.9Hz, CH 2NH), 7.99 (1H, s, CHC[morpholine]), 7.85 (1H, d, J=9.9Hz, CHCHC[morpholine]), 7.50 (1H, d, J=9.9Hz, CHCHC[morpholine]), and 7.52-7.36 (4H, m, Ar-CH), 4.50 (2H, d, J=6.9Hz, CH 2NH), 3.76 (4H, m, CH 2OCH 2) and 3.18 (4H, m, CH 2NCH 2).
MS(ESI +)m/z415(M+1)
HPLC Method 695%/7.22 minute
The preparation of embodiment 6.12:3-hydroxyl-7-morpholine-4-base-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 2-chloro-benzyl acid amides
Figure G2007800515627D00432
1H NMR (300MHz, D6-DMSO): δ 12.00 (1H, s, OH), 9.63 (1H, t, J=6.3Hz, NHCH 2), 8.00 (1H, s, CHC[morpholine]), 7.85 (1H, d, J=9.6Hz, CHCHC[morpholine]), 7.53 (1H, d, J=9.6Hz, CHCHC[morpholine]), 7.48 (1H, m, Ar-CH), 7.37-7.31 (3H, m, Ar-CH), 4.61 (2H, d, J=6.3Hz, NHCH 2), 3.78 (4H, m, CH 2OCH 2) and 3.19 (4H, m, CH 2NCH 2).
MS(ESI +)m/z415(M+1) +
HPLC Method 590%/3.85 minute
Embodiment 6.13:3-hydroxyl-7-morpholine-4-base-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
1H NMR (300MHz, D6-DMSO): δ 12.00 (1H, s, OH), 9.73 (1H, bs, CH 2NH), 7.99 (1H, s, CHC[morpholine]), 7.85 (1H, d, J=9.9Hz, CHCHC[morpholine]), 7.61 (2H, m, CHCHC[morpholine] and CHC[Cl] C[Cl]), 7.50 (2H, d, J=8.1Hz, Ar-CH), 7.35 (2H, d, J=8.1Hz, Ar-CH), 4.51 (2H, d, J=6.6Hz, CH 2NH), 3.77 (4H, m, CH 2OCH 2) and 3.18 (4H, m, CH 2NCH 2).
MS(ESI +)m/z449(M[Cl 35，Cl 35]+1)，(ESI -)m/z447(M[Cl 35，Cl 35]-1)
HPLC Method 594%/4.84 minute
The preparation of embodiment 6.14:3-hydroxyl-8-morpholine-4-base-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
Utilize the product of embodiment 2.4, adopt the program of in embodiment 6, describing (except only using 1.3 normal 4-flunamines) so that desired compounds to be provided.
1H?NMR(300MHz，D6-DMSO)：δ11.60(1H，s，OH)，9.50(1H，t，J＝6.3Hz，NH)，8.55(1H，d，J＝8.4Hz，Ar-CH)，7.41-7.36(2H，m，Ar-CH)，7.22-7.12(3H，m，Ar-CH)，6.51(1H，s，Ar-CH)，4.47(2H，d，J＝6.3Hz，NHCH 2)，3.72(4H，m，CH 2OCH 2)，3.34(4H，m，CH 2NCH 2)。
(ESI -)m/z397(M-1)
HPLC Method 794.4%/9.0 minute
Embodiment 6.15:3-hydroxyl-8-morpholine-4-base-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00451
Utilize the product of embodiment 2.4, adopt the program in embodiment 6, described (normal 3 except only using 1.3, the 4-dichloro-benzylamine) so that desired compounds to be provided.
1H?NMR(300MHz，D6-DMSO)：δ11.50(1H，s，OH)，9.61(1H，t，J＝6.3Hz，NH)，8.57(1H，d，J＝8.4Hz，Ar-CH)，7.63-7.60(2H，m，Ar-CH)，7.34(1H，d，J＝8.4Hz，Ar-CH)，7.22(1H，d，J＝8.4Hz，Ar-CH)，6.53(1H，s，Ar-CH)，4.49(2H，d，J＝6.3Hz，NHCH 2)，3.75(4H，m，CH 2OCH 2)，3.37(4H，m，CH 2NCH 2)。
(ESI -)m/z447(M[Cl 35]-1)
HPLC Method 793.2%/10.2 minute
The preparation of embodiment 6.16:3-hydroxyl-9-morpholine-4-base-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D00452
Utilize the product of embodiment 2.5, adopt the program of in embodiment 6, describing (except only using the fluorine-based benzylamine of 1.3 normal 4-) so that desired compounds to be provided.
1H?NMR(300MHz，CDCl 3)δ3.23(4H，s，-NCH 2CH 2O-)，3.76(4H，s，-NCH 2CH 2O-)，4.61(2H，d，J＝5.7Hz，-(O＝C)NHCH 2-)，6.91(2H，m，ArH)，7.09(2H，t，J＝8.4Hz，ArH)，7.34(2H，bt，ArH)，7.98(1H，s，-(O＝C)NHCH 2-)，8.61(1H，d，J＝7.2Hz，ArH)，11.80(1H，s，OH)。
(ESI +)m/z399(M+1)
HPLC Method 797.0%/11.6 minute
Embodiment 6.17:3-hydroxyl-9-morpholine-4-base-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00461
Utilize the product of embodiment 2.5, adopt the program in embodiment 6, described (normal 3,4-dichloro-benzylamine and in 1: 1 mixture of methyl alcohol/tetrahydrofuran (THF), react) so that desired compounds to be provided except only using 1.3.
1H?NMR(300MHz，CDCl 3)δ3.35(4H，s，-NCH 2CH 2O-)，3.97(4H，s，-NCH 2CH 2O-)，4.64(2H，d，J＝6.0Hz，-(O＝C)NHCH 2-)，6.99(2H，m，ArH)，7.24(1H，m，ArH)，7.48(2H，m，ArH)，8.50(1H，bs，-(O＝C)NHCH 2-)，8.69(1H，d，J＝7.8Hz，ArH)，11.84(1H，s，OH)。
(ESI +)m/z471(M+Na)
HPLC Method 791.0%/13.1 minute
Embodiment 6.18:3-hydroxyl-4-oxo-7-piperidines-l-base-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00462
Utilize the product of embodiment 2.6, adopt the program in embodiment 6, described (normal 3 except only using 1.3, the 4-dichloro-benzylamine) so that desired compounds to be provided.
1H NMR (300MHz, CDCl 3) δ 1.58 (2H, bm, ring-type-N (CH 2) 3CH 2NCH 2-), 1.73 (4H, bs, ring-type-N (CH 2) 3CH 2NCH 2-), 3.20 (4H, bm, ring-type-N (CH 2) 2CH 2NCH 2-), 4.62 (2H, d, J=6.0Hz ,-(O=C) NHCH 2-), 7.50 (4H, m, ArH), 8.24 (1H, dd, J=1.8Hz, ArH), 8.51 (1H, bs ,-(O=C) NHCH 2-), 11.86 (1H, s, OH).
(ESI -)m/z445(M[Cl 35]-1)
HPLC Method 791.0%/14.9 minute
Embodiment 7:3-cyano group-6-hydroxyl-7-oxo-l, the preparation of 7-dihydro-pyrazolo [l, 5-α] pyrimidine-5-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D00471
With 5-(4-fluoro-benzylamino formyl radical)-7-oxo-2-phenyl-1,7-dihydro-pyrazolo [1,5-a] pyrimidine-6-oleic acid ester; 4-fluoro-hexadecyldimethyl benzyl ammonium (25mg) is suspended in the water (1mL), and the interpolation aqueous hydrochloric acid (1.0M, 1mL).(surpassing) sonication mixture 5 minutes, and by the resulting throw out of filtration collection, and water (2mL) washing, use the pump drying then, to provide 3-cyano group-6-hydroxyl-7-oxo-1,7-dihydro-pyrazolo [1,5-α] pyrimidine-5-carboxylic acid 4-fluoro-benzyl acid amides (13mg) as colorless solid.
1H?NMR(300MHz，D6-DMSO)：δ4.53(2H，d，J＝5.9Hz，NHCH 2)，7.15(2H，m，ArH)，7.42(2H，m，ArH)，8.36(1H，s，H2)，7.92(2H，d，J＝7.2Hz，Ar-CH)，9.14(1H，t，J＝5.9Hz，NHCH 2)，11.25(1H，br?s，OH)
MS(ESI -)m/z326(M-1)
HPLC Method 595.4%/4.14 minute
By adopting the program of in embodiment 7, describing, obtain following compound.
Embodiment 7.1:6-hydroxyl-7-oxo-2-phenyl-l, the preparation of 7-dihydro-pyrazolo [l, 5-α] pyrimidine-5-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D00481
1H?NMR(300MHz，D6-DMSO)：δ4.58(2H，d，J＝5.9Hz，NHCH 2)，6.50(1H，s，H3)，7.15(2H，m，ArH)，7.42(5H，m，ArH)，7.93(2H，m，ArH)，9.10(1H，t，J＝5.9Hz，NHCH 2)，10.80(1H，br?s，OH)，11.84(1H，br?s，NH)
MS(ESI+)m/z379(M+1)
The preparation of embodiment 8:3-hydroxyl-7-methyl-2-(tolyl between 5--[l, 3,4] oxadiazole-2-yl)-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D00482
The preparation of embodiment 8.1:3-benzyloxy-7-methyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylate methyl ester
Figure G2007800515627D00483
At N 2Under the atmosphere, will from the product of embodiment 2.1 (1.5g, 6.4mmol) and salt of wormwood (2.7g 19.6mmol) mixes with acetone (30mL).Under 70 ℃, stirred the mixture 25 minutes, add afterwards bromotoluene (2.0g, 11.7mmol) and the mixture 10 hours of refluxing.After being cooled to room temperature, mixture is injected in the water (100mL), use dichloromethane extraction.Wash organic phase with water, dry and concentrated in a vacuum.The purifying that is undertaken by rapid column chromatography method (methylene dichloride) provides desired compounds (1.5g, 70%).
1H?NMR(300MHz，CDCl 3)：δ2.44(d，J＝0.9Hz，3H)，3.92(s，3H)，5.32(s，2H)，7.27-7.41(m，3H)，7.47-7.57(m，3H)，7.65(d，J＝9.1Hz，1H)，8.76-8.85(m，1H)。
MS(ESI +)m/z?325(M+1)，347(M+23)。
The preparation of embodiment 8.2:3-benzyloxy-7-methyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid
Figure G2007800515627D00491
At room temperature, (400mg adds 1N lithium hydroxide aqueous solution (2.46mL) in stirred solution 1.23mmol) to the product in methyl alcohol (20mL) from embodiment 8.1.After 3 hours, add 1N aqueous hydrochloric acid (20mL).Use the ethyl acetate extraction mixture, and with salt water washing organic phase, dry (Na 2SO 4) and under vacuum, concentrate.Product is directly used in embodiment 8.3.
The preparation of embodiment 8.3:3-methyl-phenylformic acid N '-(3-benzyloxy-7-methyl-4-oxygen-4H-pyrido [l, 2-α] pyrimidine-2-carbonyl)-hydrazides
Figure G2007800515627D00492
At room temperature to the product (200mg in tetrahydrofuran (THF) (10mL) from embodiment 8.2,0.644mmol) solution in add 3-toluyl hydrazine (94.8mg successively, 0.632mmol), I-hydroxybenzotriazole (6.98mg, 0.0576mmol) and 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (98mg, 0.632mmol).After 12 hours, water (20mL) quenching reaction soln is also used ethyl acetate extraction.With 2N aqueous hydrochloric acid (20mL), 2N aqueous NaOH (20mL) washing extract, dry (Na 2SO 4) and under vacuum, concentrate, so that desired compounds (53%) to be provided.
1H?NMR(300MHz，DMSO-d 6)：δ2.39(s，3H)，2.44(s，3H)，5.21(s，2H)，7.30-7.45(m，5H)，7.56-7.63(m，2H)，7.69-7.79(m，3H)，7.84(dd，J＝9.4，2.1Hz，1H)，8.78-8.85(m，1H)，10.56(d，J＝11.1Hz，2H)。
MS(ESI +)m/z443(M+1)，465(M+23)。
The preparation of embodiment 8.4:3-benzyloxy-7-methyl-2-(tolyl between 5--[l, 3,4] oxadiazole-2-yl)-pyrido [1,2-α] pyrimidin-4-one
Figure G2007800515627D00501
Will from the product of embodiment 8.3 (202mg, 0.457mmol), tetracol phenixin (0.221mL, 2.28mmol) and triethylamine (0.165mL 1.19mmol) mixes with acetonitrile (10mL).At room temperature in this mixture, add triphenylphosphine (291mg, 1.11mmol).After at room temperature stirring is spent the night, with ethyl acetate (100mL) diluting reaction solution, use moisture saturated sodium bicarbonate (50mL), water (50mL) and salt solution (50mL) washing successively, dry then (Na 2SO 4).Crude product is carried out flash chromatography (hexane-ethyl acetate 1: 1) to provide desired compounds.
1H?NMR(300MHz，DMSO-d 6)：δ2.41(s，3H)，2.46(d，J＝1.1Hz，3H)，5.32(s，2H)，7.28-7.33(m，3H)，7.46-7.52(m，4H)，7.75-7.90(m，4H)，8.82-8.86(m，1H)。
MS(ESI +)m/z?425(M+1)，447(M+23)。
The preparation of embodiment 8.5:3-hydroxyl-7-methyl-2-(tolyl between 5--[l, 3,4] oxadiazole-2-yl)-pyrido [1,2-α] pyrimidin-4-one
At N 2Under room temperature, to from the product in acetonitrile (5mL) of embodiment 8.4 (20mg, drip in stirred solution 0.047mmol) Iodotrimethylsilane (54 μ L, 0.38mmol).After 2 hours, added methyl alcohol (5mL) and stirred solution 10 minutes.Add water (10mL) and react with dichloromethane extraction.Wash organic phase with aqueous solution of sodium bisulfite, dry (Na 2SO 4) and under vacuum, concentrate, so that desired compounds (88.6%) to be provided.
1H?NMR(300MHz，CDCl 3)：δ2.43(s，3H)，2.49(s，3H)，7.38-7.50(m，3H)，7.65(d，J＝8.8Hz，1H)，8.03-8.09(m，2H)，8.78(s，1H)，9.92(brs，1H)。
MS(ESI +)m/z?335(M+1)，357(M+23)。
The preparation of embodiment 8.6:3-hydroxyl-7-methyl-2-(5-phenyl-[1,3,4] oxadiazole-2-yls)-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D00512
The program that employing is described in embodiment 8.1-8.5 prepares 3-hydroxyl-7-methyl-2-(5-phenyl-[1,3,4] oxadiazole-2-yls)-pyrido [1,2-α] pyrimidin-4-one.
1H?NMR(300MHz，CDCl 3)δ2.43(s，3H)，7.45(d，J＝8.5Hz，1H)，7.54-7.68(m，4H)，8.26(d，J＝6.6Hz，2H)，8.77(s，1H)
MS(ESI +)m/z?321(M+1)
HPLC Method 782.8%/14.3 minute
The preparation of embodiment 8.7:2-[5-(2-chloro-phenyl)-[l, 3,4] oxadiazole-2-yl]-3-hydroxyl-7-methyl-pyrido [1,2-α] pyrimidin-4-one
The program that employing is described in embodiment 8.1-8.5 prepares 2-[5-(2-chloro-phenyl)-[1,3,4] oxadiazole-2-yl]-3-hydroxyl-7-methyl-pyrido [1,2-α] pyrimidin-4-one.
1H?NMR(300MHz，CDCl 3)δ2.43(d，J＝1.1Hz，3H)，7.44(dd，J＝9.2，2.2Hz，1H)，7.49(dd，J＝7.7，1.5Hz，1H)，7.55(dt，J＝7.5，1.8Hz，1H)，7.60-7.66(m，2H)，8.13(dd，J＝7.7，1.8Hz，1H)，8.76-8.79(m，1H)，9.71-9.91(brs，1H)
MS(ESI +)m/z377(M+Na +)
HPLC Method 792.2%/15.4 minute
The preparation of embodiment 8.8:2-[5-(4-methoxyl group-phenyl)-[1,3,4] oxadiazole-2-yl]-3-hydroxyl-7-methyl-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D00522
The program that employing is described in embodiment 8.1-8.5 prepares 2-[5-(4-methoxyl group-phenyl)-[1,3,4] oxadiazole-2-yl]-3-hydroxyl-7-methyl-pyrido [1,2-α] pyrimidin-4-one.
1H?NMR(300MHz，CDCl 3)δ2.43(s，3H)，3.92(s，3H)，7.07(d，J＝8.9Hz，2H)，7.44(dd，J＝9.5Hz，1.9Hz，1H)，7.64(d，J＝9.4Hz，1H)，8.21(d，J＝8.9Hz，2H)，8.78(d，J＝1.9Hz，1H)，9.88-10.10(brs，1H)
MS(ESI +)m/z373(M+Na +)
HPLC Method 792.4%/15.3 minute
The preparation of embodiment 8.9:2-[5-(4-fluoro-phenyl)-[l, 3,4] oxadiazole-2-yl]-3-hydroxyl-7-morpholine-4-base-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D00531
Be used to product from embodiment 2.3 as starting raw material, adopt the program of in embodiment 8.1-8.5, describing to prepare 2-[5-(4-fluoro-phenyl)-[1,3,4] oxadiazole-2-yl]-3-hydroxyl-7-morpholine-4-base-pyrido [1,2-α] pyrimidin-4-one.
1H?NMR(300MHz，DMSO-d 6)δ3.23(t，J＝4.8Hz，4H)，3.80(t，J＝4.8Hz，4H)，7.52(t，J＝9.0Hz，2H)，7.67(d，J＝10.0Hz，1H)，7.87(dd，J＝10.0Hz，2.4Hz，1H)，8.03(d，J＝2.3Hz，1H)，8.16(dd，J＝8.8Hz，5.1Hz，2H)，10.46-10.60(brs，1H)
HPLC Method 798.4%/8.5 minute
The preparation of embodiment 9:2-[5-(4-fluoro-benzyl)-[l, 3,4] oxadiazole-2-yl]-3-hydroxyl-7-methyl-pyrido [l, 2-a] pyrimidin-4-one
Figure G2007800515627D00532
The preparation of embodiment 9.1:3-benzyloxy-7-methyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid hydrazides
Figure G2007800515627D00533
At room temperature (800mg adds hydrazine (6.0 equivalent) in stirred solution 2.56mmol) to the product in methyl alcohol (30mL) from embodiment 8.1.45 ℃ of following heated mixt 4 hours, then partly under vacuum, concentrate (not being concentrated into drying) then, be cooled to room temperature then.Filter resulting solid, wash with water and dry under vacuum, so that desired compounds (650mg, productive rate 78%) to be provided.
1H?NMR(300MHz，DMSO-d 6)：δ2.42(s，3H)，5.15(s，2H)，7.28-7.45(m，3H)，7.48-7.53(m，2H)，7.66(d，J＝8.8Hz，1H)，7.80(dd，J＝8.8，2.1Hz，1H)，8.78(s，1H)，8.93(brs，2H)，9.7(brs，1H)。
MS(ESI +)m/z?325(M+1)，347(M+23)。
The preparation of embodiment 9.2:3-benzyloxy-7-methyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid N '-[2-(4-fluoro-phenyl)-ethanoyl]-hydrazides
Figure G2007800515627D00541
Will from the product of embodiment 9.1 (160mg, 0.524mmol) and yellow soda ash (106mg 1mmol) mixes with anhydrous tetrahydro furan (25mL), cools off in ice bath then.The fluorine-based phenyl Acetyl Chloride 98Min. of dropping 4-in this stirred solution (90mg, 0.55mmol).At room temperature stirred the mixture 2 hours, and under vacuum, concentrated then.Between ethyl acetate and water, distribute residue, and wash organic phase with water, dry (Na 2SO 4) and under vacuum, concentrate.The short column chromatography provides desired compounds (210mg, productive rate 86%).
1H?NMR(300MHz，CDCl 3)：δ2.45(s，3H)，3.67(s，2H)，5.40(s，2H)，6.95-7.10(m，2H)，7.30-7.42(m，5H)，7.50-7.60(m，3H)，7.64(d，J＝9.4Hz，1H)，8.70-8.80(m，2H)，10.42(brs，1H)。
MS(ESI +)m/z?461(M+1)，483(M+23)。
The preparation of embodiment 9.3:3-benzyloxy-2-[5-(4-fluoro-benzyl)-[l, 3,4] oxadiazole-2-yl]-7-methyl-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D00551
The program that to describe in embodiment 8.4 is used for the product (70%) of product so that expectation to be provided in embodiment 9.2 acquisitions.
1H?NMR(300MHz，CDCl 3)：δ2.46(d，J＝0.9Hz，3H)，4.25(s，2H)，5.39(s，2H)，6.96-7.05(m，2H)，7.27-7.34(m，5H)，7.38-7.45(m，2H)，7.56(dd，J＝9.1Hz，2.0Hz，1H)，7.70(d，J＝9.1Hz，1H)，8.78-8.83(m，1H)。
MS(ESI +)m/z443(M+1)，465(M+23)。
The preparation of embodiment 9.4:2-[5-(4-fluoro-benzyl)-[l, 3,4] oxadiazole-2-yl]-3-hydroxyl-7-methyl-pyrido [l, 2-a] pyrimidin-4-one
Figure G2007800515627D00552
The program that to describe in embodiment 8.5 is used for the product (52%) of product so that expectation to be provided in embodiment 9.3 acquisitions.
1H?NMR(300MHz，CDCl 3)：δ2.42(s，3H)，4.37(s，2H)，7.02-7.11(m，2H)，7.32-7.48(m，3H)，7.61(d，J＝9.6Hz，1H)，8.76(s，1H)，9.79(brs，1H)。
MS(ESI -)m/z351(M-1)
HPLC Method 797.3%/8.5 minute
Embodiment 9.5:2-[5-(3,4-two chloro-benzyls)-[l, 3,4] oxadiazole-2-yl]-preparation of 3-hydroxyl-7-methyl-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D00561
The program that employing is described in embodiment 9.1-9.4 prepares 2-[5-(3,4-two chloro-benzyls)-[1,3,4] oxadiazole-2-yl]-3-hydroxyl-7-methyl-pyrido [1,2-α] pyrimidin-4-one.
1H NMR (300MHz, CDCl 3) δ 2.43 (s, 3H), 4.35 (s, 2H), 7.26 (1H, overlapping), 7.41-7.55 (m, 3H), 7.64 (d, J=9.2Hz, 1H), 8.76 (s, 1H), 9.55-9.85 (brs, 1H)
MS(ESI -)m/z401(M-1)
HPLC Method 797.6%/18.0 minute
Embodiment 9.6:2-[5-(3,4-two chloro-benzyls)-[l, 3,4] oxadiazole-2-yl]-preparation of 3-hydroxyl-7-morpholine-4-base-pyrido [l, 2-a] pyrimidin-4-one
The starting raw material that utilization prepares in embodiment 2.3 adopts the program of describing in embodiment 9.1-9.4 to prepare 2-[5-(3,4-two chloro-benzyls)-[1,3,4] oxadiazole-2-yl]-3-hydroxyl-7-methyl-pyrido [1,2-α] pyrimidin-4-one.
1H?NMR(300MHz，DMSO-d 6)δ3.18-3.24(m，4H)，3.75-3.83(m，4H)，4.47(s，2H)，7.40(dd，J＝8.3，2.0Hz，1H)，7.61(d，J＝9.8Hz，1H)，7.65(d，J＝8.5Hz，1H)，7.72(d，J＝2.0Hz，1H)，7.85(dd，J＝9.9，2.5Hz，1H)，8.01(d，J＝2.5Hz，1H)，10.41(s，1H)
HPLC Method 794.1%/17.2 minute
The preparation of embodiment 9.7:2-[5-(4-fluoro-benzyl)-[l, 3,4] oxadiazole-2-yl]-3-hydroxyl-7-morpholine-4-base-pyrido [l, 2-α] pyrimidin-4-one
The starting raw material that utilization prepares in embodiment 2.3 adopts the program of describing in embodiment 9.1-9.4 to prepare 2-[5-(4-fluoro-benzyl)-[1,3,4] oxadiazole-2-yl]-3-hydroxyl-7-methyl-pyrido [1,2-α] pyrimidin-4-one.
1H?NMR(300MHz，DMSO-d 6)δ3.20(t，J＝4.8Hz，4H)，3.79(t，J＝4.8Hz，4H)，4.42(s，2H)，7.21(t，J＝9.0Hz，2H)，7.43(dd，J＝8.8，5.5Hz，2H)，7.61(d，J＝9.9Hz，1H)，7.85(dd，J＝9.8，2.5Hz，1H)，8.00(d，J＝2.5Hz，1H)，10.39(s，1H)。
MS(ESI -)m/z?422(M-1)
HPLC Method 794.1%/14.7 minute
Embodiment 10:2-[5-(4-fluoro-benzyl)-[l, 3,4] thiadiazoles-2-yl]-preparation of 3-hydroxyl-7-methyl-pyrido [l, 2-a] pyrimidin-4-one
Figure G2007800515627D00572
Embodiment 10.1:3-benzyloxy-2-[5-(4-fluoro-benzyl)-[l, 3,4] thiadiazoles-2-yl]-preparation of 7-methyl-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D00581
Will from the product of embodiment 9.2 (80mg, 0.173mmol) and La Weisong reagent (Lawensson ' s reagent) (200mg 0.5mmol) mixes with toluene (15mL) and refluxed 10 hours.Concentrated reaction mixture under vacuum, flash chromatography (ethyl acetate/dichloromethane/ether 2: 6: 1) provides desired compounds (60mg, 75.3%) then.
1H?NMR(300MHz，DMSO-d 6)：δ2.42(s，3H)，4.53(s，2H)，5.26(s，2H)，7.17-7.26(m，2H)，7.29-7.35(m，3H)，7.40-7.49(m，4H)，7.67(d，J＝9.5Hz，1H)，7.80(dd，J＝9.4Hz，1.9Hz，1H)，8.77(d，J＝1.1Hz，1H)。
MS(ESI +)m/z459(M+1)，481(M+23)。
Embodiment 10.2:2-[5-(4-fluoro-benzyl)-[l, 3,4] thiadiazoles-2-yl]-preparation of 3-hydroxyl-7-methyl-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D00582
The program of describing in embodiment 8.5 is used for the product (34%) of product so that expectation to be provided in embodiment 10.1 acquisitions
1H?NMR(300MHz，CDCl 3)：δ2.39(d，J＝0.8Hz，3H)，4.48(s，2H)，7.02-7.12(m，2H)，7.30-7.45(m，4H)，8.71-8.77(m，1H)，10.80(brs，1H)。
MS(ESI +)m/z369(M+1)，391(M+23)。
HPLC Method 796.7%/15.8 minute
Embodiment 10.3:2-[5-(3,4-two chloro-benzyls)-[l, 3,4] thiadiazoles-2-yl]-preparation of 3-hydroxyl-7-methyl-pyrido [1,2-α] pyrimidin-4-one
Figure G2007800515627D00591
The program that employing is described in embodiment 10.1-10.2 prepares 2-[5-(3,4-two chloro-benzyls)-[1,3,4] thiadiazoles-2-yl]-3-hydroxyl-7-methyl-pyrido [1,2-α] pyrimidin-4-one
1H?NMR(300MHz，CDCl 3)δ3.16-3.25(m，4H)，3.73-3.83(m，4H)，4.29(s，2H)，7.37(d，J＝8.6Hz，1H)，7.63(d，J＝8.3Hz，2H)，7.69(s，1H)，7.86(d，J＝8.8Hz，1H)，7.98(s，1H)，10.71(s，1H)
MS(ESI -)m/z417(M[Cr 35]-1)
HPLC Method 797.8%/19.8 minute
Embodiment 10.4:2-[5-(3,4-two chloro-benzyls)-[l, 3,4] thiadiazoles-2-yl]-preparation of 3-hydroxyl-7-morpholine-4-base-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D00592
The starting raw material that utilization prepares in embodiment 2.3 adopts the program of describing in embodiment 10.1-10.2 to prepare 2-[5-(3,4-two chloro-benzyls)-[1,3,4] thiadiazoles-2-yl]-3-hydroxyl-7-morpholine-4-base-pyrido [1,2-α] pyrimidin-4-one
1H?NMR(300MHz，DMSO-d 6)δ3.15-3.25(m，4H)，3.70-3.85(m，4H)，4.60(s，2H)，7.42(dd，J＝8.2Hz，2.1Hz，1H)，7.60(d，J＝9.8Hz，1H)，7.65(d，J＝8.2Hz，1H)，7.74(d，J＝2.1Hz，1H)，7.86(dd，J＝9.8Hz，2.5Hz，1H)，8.01(d，J＝2.3Hz，1H)，10.50-11.10(brs，1H)
MS(ESI -)m/z488(M-1)
HPLC Method 797.6%/19.3 minute
Embodiment 10.5:2-[5-(4-fluoro-benzyl)-[l, 3,4] thiadiazoles-2-yl]-preparation of 3-hydroxyl-7-morpholine-4-base-pyrido [1,2-α] pyrimidin-4-one
The starting raw material that utilization prepares in embodiment 2.3 adopts the program of describing among the embodiment 9.1-9.4 to prepare 2-[5-(4-fluoro-benzyl)-[1,3,4] thiadiazoles-2-yl]-3-hydroxyl-7-morpholine-4-base-pyrido [1,2-α] pyrimidin-4-one
1H?NMR(300MHz，DMSO-d 6)δ3.20(t，J＝4.8Hz，4H)，3.78(t，J＝4.8Hz，4H)，4.56(s，2H)，7.21(t，J＝8.8Hz，2H)，7.47(dd，J＝8.8Hz，5.5Hz，2H)，7.59(d，J＝9.8Hz，1H)，7.85(dd，J＝9.9Hz，2.7Hz，1H)，8.01(d，J＝2.6Hz，1H)，10.80(s，1H)
MS(ESI -)m/z438(M-1)
HPLC Method 794.1%/14.2 minute
The preparation of embodiment 11:2-[5-(4-fluoro-benzyl)-[l, 2,4] oxadiazole-3-yl]-3-hydroxyl-7-methyl-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D00602
The preparation of embodiment 11.1:3-benzyloxy-7-methyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-formoxime (carbaldehyde oxime)
Will (3.1g 10mmol) be dissolved in the anhydrous tetrahydro furan (50mL) and be cooled to-78 ℃ from the product of embodiment 8.1.In this stirred solution, drip diisobutylaluminium hydride (13mL, 1N is in tetrahydrofuran (THF)).After 4 hours, TLC shows that starting raw material is consumed, and with aqueous sodium persulfate solution quenching reaction soln.Leach insoluble substance and concentrated filtrate under vacuum.Residue is dissolved in ethyl acetate/dichloromethane (1: in mixed solvent 15mL), and use the salt water washing, dry (Na 2SO 4) and filter.
Oxammonium hydrochloride among the Xiang Zaishui (120mL) (760mg adds above-mentioned aldehyde solution in solution 11mmol), then add sodium bicarbonate (900mg, 10.7mmol).At room temperature stirring the mixture 2 hours, and by filter collecting resulting throw out, washing with water then and dry under vacuum, with the product (2.77g, two step overall yields 90%) that expectation is provided.
The preparation of embodiment 11.2:3-benzyloxy-7-methyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-nitrile
Figure G2007800515627D00611
With three chloro-1,3,5-triazines (576mg 3.15mmol) is dissolved in anhydrous N, dinethylformamide (DMF) (1mL) in and at room temperature stirred 30 minutes.In this solution, drip the product from embodiment 11.1 (927mg, solution 3mmol) in DMF (5mL).Mixture was kept at room temperature 2 hours, add ethyl acetate (50mL) then, separate organic phase then and use the salt water washing, dry (Na 2SO 4) and under vacuum, concentrate.Carry out purifying so that desired compounds (530mg, 60.7%) to be provided by short flash chromatography.
1H?NMR(300MHz，CDCl 3)：δ2.45(d，J＝I.2Hz，3H)，5.54(s，2H)，7.30-7.40(m，3H)，7.48-7.54(m，2H)，7.55-7.58(m，2H)，8.77(dd，J＝2.7，1.2Hz，1H)。
The preparation of embodiment 11.3:3-benzyloxy-N-hydroxyl-7-methyl-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carbonamidine (carboxamidine)
Figure G2007800515627D00621
Will from the product of embodiment 11.2 (530mg, 1.82mmol) and oxammonium hydrochloride (0.549g 7.9mmol) mixes with ethanol (50mL).(663mg is 7.9mmol) and 70 ℃ of following heated mixt 3 hours to add sodium bicarbonate in this stirred solution.Under vacuum, remove and desolvate and residue is dissolved in mixed solvent (methylene dichloride/ethanol 200mL: 10mL), wash dry (Na with water 2SO 4) and under vacuum, concentrate, to provide desired compounds (472mg, 80%).
MS(ESI +)m/z?325(M+1)，347(M+23)，379(M+55)。
The preparation of embodiment 11.4:3-benzyloxy-2-[5-(4-fluoro-benzyl)-[l, 2,4] oxadiazole-3-yl]-7-methyl-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D00622
Under agitation, will (472mg 1.46mmol) be dissolved in methylene dichloride/tetrahydrofuran (THF) (120mL: in mixed solvent 120mL) from the product of embodiment 11.3.Add then triethylamine (155mg, 1.53mmol), then drip 4-fluorophenyl Acetyl Chloride 98Min. (263mg, 1.53mmol).At room temperature stirred the mixture 2 hours, and under vacuum, concentrated then and the residue of gained is dissolved in the ethyl acetate, wash dry (Na then with water 2SO 4) and under vacuum, concentrate.Do not having to use resulting solid under the situation of purifying.
Be suspended in above-mentioned solid (668mg) in the toluene (25mL) and the mixture 24 hours of refluxing.Concentrated solvent is quantitatively to provide desired compounds under vacuum.
1H?NMR(300MHz，CDCl 3)：δ2.46(d，J＝0.7Hz，3H)，4.32(s，2H)，5.38(s，2H)，7.00-7.08(m，2H)，7.20-7.30(m，3H)，7.32-7.37(m，2H)，7.44-7.52(m，2H)，7.55(dd，J＝9.2，1.9Hz，1H)，7.74(d，J＝9.1Hz，1H)，8.81-8.85(m，1H)。
MS(ESI +)m/z443(M+1)，465(M+23)。
The preparation of embodiment 11.5:2-[5-(4-fluoro-benzyl)-[l, 2,4] oxadiazole-3-yl]-3-hydroxyl-7-methyl-pyrido [1,2-α] pyrimidin-4-one
Figure G2007800515627D00631
The program of describing in embodiment 8.5 is used for the product (68%) of product so that expectation to be provided in embodiment 11.4 acquisitions.
1H?NMR(300MHz，CDCl 3)：δ2.41(d，J＝0.7Hz，3H)，4.39(s，2H)，7.01-7.12(m，2H)，7.32-7.44(m，3H)，7.68(d，J＝9.2Hz，1H)，8.70(s，1H)，8.72-8.90(brs，1H)。
MS(ESI +)m/z353(M+1)，375(M+23)。
HPLC Method 794.5%/14.4 minute
The preparation of embodiment 12:3-hydroxyl-7-methyl-2-(5-phenyl-oxazoles-2-yl)-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D00632
The preparation of embodiment 12.1:2-amino-l-phenyl-ethyl ketone (ethanone)
Figure G2007800515627D00641
Blend alpha in ethanol (425mL)-bromo methyl phenyl ketone (7.0g, 0.035mol), vulkacit H (5.4g, 0.0385mol) and sodium iodide (5.8g 0.0385mol), at room temperature stirred 24 hours then.Filter reaction mixture, and with cold washing with alcohol filter cake, the solid with gained is dissolved in the ethanol (100mL) then, and add 6N aqueous hydrochloric acid (20mL).Backflow mixture 5 hours is cooled to room temperature then.Filtering mixt and under vacuum concentrated filtrate.The residue recrystallization of gained is from the product (4.1g, 69%) of diisopropyl ether/concentrated hydrochloric acid (100/1) so that expectation to be provided.
1H?NMR(300MHz，CDCl 3)：δ4.57(s，2H)，7.56-7.691(m，2H)，7.71-7.76(m，1H)，8.00-8.03(m，2H)，8.52(br?s，3H)。
The preparation of embodiment 12.2:3-benzyloxy-7-methyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid (2-oxo-2-phenyl-ethyl)-acid amides
Figure G2007800515627D00642
At room temperature, to the product (324mg in tetrahydrofuran (THF) (15mL) from embodiment 8.2, add product (162mg in stirred solution 1mmol) successively from embodiment 12.1,1.2mmol), I-hydroxybenzotriazole (162mg, 1.2mmol), 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (191mg, 1mmol) and triethylamine (112mg, 1.1mmol).After 3 hours, with saturated sodium bicarbonate aqueous solution (5mL) quenching reaction soln.Use the ethyl acetate extraction mixture, and water, salt water washing organic phase, (Na used then 2SO 4) dry and concentrated under vacuum.The flash chromatography of residue provides the product (215mg, 45%) of expectation.
1H?NMR(300MHz，CDCl 3)：δ2.45(s，3H)，4.93(d，J＝4.5Hz，2H)，5.44(s，2H)，7.26-7.32(m，3H)，7.51-7.72(m，7H)，8.03(d，J＝7.5Hz，2H)，8.65(s，1H，NH)，8.79(s，1H)。
MS(ESI +)m/z?428(M+1)，450(M+Na +)，482(M+MeOH+Na +)。
The preparation of embodiment 12.3:3-benzyloxy-7-methyl-2-(5-phenyl-oxazoles-2-yl)-pyrido [1,2-a] pyrimidin-4-one
Figure G2007800515627D00651
At room temperature, to from the product in acetonitrile (5mL) of embodiment 12.2 (170mg, add successively in stirred solution 0.4mmol) tetracol phenixin (360mg, 2.4mmol), triethylamine (130mg, 1.28mmol) and triphenylphosphine (320mg, 1.2mmol).After 2 hours, add saturated sodium bicarbonate aqueous solution (5mL) and use the ethyl acetate extraction product.Water, salt water washing organic phase, dry then (Na 2SO 4) and under vacuum, concentrate.The flash chromatography of residue provides desired compounds (142mg, 86%).
1H?NMR(300MHz，CDCl 3)δ2.45(s，3H)，5.46(s，2H)，7.29-7.37(m，6H)，7.54-7.61(m，6H)，7.75(d，1H)，8.80(s，1H)。
MS(ESI -)m/z?380(M-1)；MS(ESI +)m/z410(M+1)，432(M+Na +)，464(M+MeOH+Na +)，841(2M+Na +)。
The preparation of embodiment 12.4:3-hydroxyl-7-methyl-2-(5-phenyl-oxazoles-2-yl)-pyrido [1,2-α] pyrimidin-4-one
Figure G2007800515627D00652
Make product from embodiment 12.3 (62mg, 0.5mmol) and sodium iodide (440mg 2.9mmol) mixes with acetonitrile (5mL).In this stirred solution, drip trimethylchlorosilane (316mg, 2.9mmol).Stirred the mixture 1 hour, and by adding methyl alcohol (5mL), then adding in addition quenching of water (20mL), used ethyl acetate extraction then then.With the organic layer that the salt water washing merges, dry (Na 2SO 4) and under vacuum, be concentrated into the volume of 1mL.Drip hexane (15mL) and collect resulting solid by filtering, dry under vacuum then, with the product (38mg, 79%) that provides expectation.
1H?NMR(300MHz，CDCl 3)δ2.40(s，3H)，7.40-7.57(m，6H)，7.84(s，2H)，8.74(s，1H)。
MS(ESI +)m/z?320(M+1)，342(M+Na +)，374(M+MeOH+Na +)，661(2M+Na +)。
HPLC Method 795.0%/15.6 minute
Embodiment 12.5:2-[5-(4-fluoro-phenyl)-oxazoles-2-yl]-preparation of 3-hydroxyl-7-methyl-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D00661
The program that employing is described in embodiment 12.1-12.4 prepares 2-[5-(4-fluoro-phenyl)-oxazoles-2-yl]-3-hydroxyl-7-methyl-pyrido [1,2-α] pyrimidin-4-one.
1H?NMR(300MHz，CDCl 3)δ2.43(s，3H)，7.45(d，J＝8.5Hz，1H)，7.54-7.68(m，4H)，8.26(d，J＝6.6Hz，2H)，8.77(s，1H)
MS(ESI +)m/z?321(M+1)
HPLC Method 782.8%/15.5 minute
Embodiment 12.6:2-[5-(4-methoxyl group-phenyl)-oxazoles-2-yl]-preparation of 3-hydroxyl-7-methyl-pyrido [l, 2-α] pyrimidin-4-one
The program that employing is described in embodiment 12.1-12.4 prepares 2-[5-(4-methoxyl group-phenyl)-oxazoles-2-yl]-3-hydroxyl-7-methyl-pyrido [1,2-α] pyrimidin-4-one.
1H?NMR(300MHz，CDCl 3)δ2.40(s，3H)，3.87(s，3H)，6.99(d，J＝8.3Hz，2H)，7.39(d，J＝9.2Hz，1H)，7.45(s，1H)，7.62(d，J＝9.2Hz，1H)，7.77(d，J＝8.5Hz，2H)，8.75(s，1H)，10.75-11.35(brs，1H)
MS(ESI +)m/z?372(M+Na +)
HPLC Method 794.1%/16.1 minute
The preparation of embodiment 12.7:l-amino-3-(4-fluoro-phenyl)-third-2-keto hydrochloride
Figure G2007800515627D00671
Adopt as at Tetrahedron.1994,50 (21), 6287-6298 and Chem.Pharm.Bull.1984,32 (7), the program described in the 2536-2543 is to provide 1-amino-3-(4-fluoro-phenyl)-third-2-keto hydrochloride.
The preparation of embodiment 12.8:l-amino-3-(3,4-two chloro-phenyl)-third-2-keto hydrochloride
Figure G2007800515627D00672
Adopt as at Tetrahedron.1994,50 (21), 6287-6298 and Chem.Pharm.Bull.1984,32 (7), the program described in the 2536-2543 is to provide 1-amino-3-(3,4-two chloro-phenyl)-third-2-keto hydrochloride.
Embodiment 12.9:2-[5-(4-fluoro-benzyl)-oxazoles-2-yl]-preparation of 3-hydroxyl-7-methyl-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D00681
Be used to from the material of embodiment 12.7 and adopt the program of embodiment 12.2-4,2-[5-(4-fluoro-benzyl)-oxazoles-2-yl are provided]-3-hydroxyl-7-methyl-pyrido [1,2-α] pyrimidin-4-one.
1H?NMR(300MHz，CDCl 3)δ2.41(s，3H)，4.17(s，2H)，6.81-7.18(m，3H)，7.26-7.60(m，4H)，8.77(s，1H)，10.40-11.80(brs，1H)
MS(ESI +)m/z?352(M+1)
HPLC Method 789.6%/15.5 minute
The preparation of embodiment 13:3-hydroxyl-7-methyl-2-(5-phenyl-thiazol-2-yl)-pyrido [1,2 α] pyrimidin-4-one
Figure G2007800515627D00682
The preparation of embodiment 13.1:3-benzyloxy-7-methyl-2-(5-phenyl-thiazol-2-yl)-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D00683
Will from the product of embodiment 12.2 (100mg, 0.23mmol) and La Weisong reagent (120mg 0.3mmol) mixes with toluene (10mL) and refluxed 12 hours.Concentrated reaction mixture and flash chromatography provide desired compounds (27mg, productive rate 27%) under vacuum.
1H?NMR(300MHz，CDCl 3)：δ2.44(s，3H)，5.55(s，2H)，7.30-7.70(m，11H)，7.80(d，J＝9.2Hz，1H)，8.28(s，1H)，8.80(s，1H)。
The preparation of embodiment 13.2:3-hydroxyl-7-methyl-2-(5-phenyl-thiazol-2-yl)-pyrido [1,2 α] pyrimidin-4-one
The program of describing in embodiment 8.5 is used for the product (80%) of product so that expectation to be provided in embodiment 13.1 acquisitions.
1H?NMR(300MHz，CDCl 3)：δ2.39(s，3H)，7.32-7.56(m，5H)，7.62-7.70(m，2H)，8.12(s，1H)，8.75(s，1H)，11.65(brs，1H)。
MS(ESI +)m/z?336(M+1)
HPLC Method 798.7%/17.5 minute
Embodiment 13.3:2-[5-(4-fluoro-benzyl)-thiazol-2-yl]-preparation of 3-hydroxyl-7-methyl-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D00692
Be used to from the material of embodiment 12.7 and adopt the program of embodiment 13.1 to 13.2,2-[5-(4-fluoro-benzyl)-thiazol-2-yl is provided]-3-hydroxyl-7-methyl-pyrido [1,2-α] pyrimidin-4-one.
1H?NMR(300MHz，DMSO-d 6)δ2.35(d，J＝I.2Hz，3H)，4.31(s，2H)，7.18(t，J＝9.9Hz，2H)，7.39(dd，J＝8.9Hz，5.5Hz，2H)，7.45-7.60(m，2H)，7.95(s，1H)，8.58-8.64(m，1H)，11.31(s，1H)
MS(ESI +)m/z390(M+Na +)
HPLC Method 796.7%/18.5 minute
Embodiment 13.4:2-[5-(3,4-two chloro-benzyls)-thiazol-2-yl]-preparation of 3-hydroxyl-7-methyl-pyrido [1,2-α] pyrimidin-4-one
Figure G2007800515627D00701
Be used to from the material of embodiment 12.8 and adopt the program of embodiment 13.1 to 13.2,2-[5-(3,4-two chloro-benzyls)-thiazol-2-yl is provided]-3-hydroxyl-7-methyl-pyrido [1,2-α] pyrimidin-4-one.
1H?NMR(300MHz，DMSO-d 6)δ2.36(d，J＝1.2Hz，3H)，4.34(s，2H)，7.37(dd，J＝8.3Hz，2.0Hz，1H)，7.47-7.60(m，2H)，7.62(d，J＝8.2Hz，1H)，7.67(d，J＝2.1Hz，1H)，7.98(s，1H)，8.60-8.65(m，1H)，11.28(s，1H)
MS(ESI +)m/z418(M+1)
HPLC Method 798.8%/19.8 minute
Embodiment 13.5:2-[5-(4-fluoro-benzyl)-thiazol-2-yl]-preparation of 3-hydroxyl-7-morpholine-4-base-pyrido [1,2-α] pyrimidin-4-one
Figure G2007800515627D00702
Be used to from the material of embodiment 2.3 and embodiment 12.7 and adopt the program of embodiment 13.1 to 13.2,2-[5-(4-fluoro-benzyl)-thiazol-2-yl is provided]-3-hydroxyl-7-morpholine-4-base-pyrido [1,2-α] pyrimidin-4-one.
1H?NMR(300MHz，DMSO-d 6)δ3.14-3.21(m，4H)，3.74-3.81(m，4H)，4.31(s，2H)，7.18(t，J＝8.9Hz，2H)，7.39(dd，J＝8.8Hz，5.5Hz，2H)，7.53(d，J＝9.9Hz，1H)，7.83(dd，J＝9.9Hz，2.6Hz，1H)，7.95(s，1H)，8.04(d，J＝2.5Hz，1H)，11.25(s，1H)
MS(ESI +)m/z461(M+Na +)
HPLC Method 786.3%/19.6 minute
Embodiment 13.6:2-[5-(3,4-two chloro-benzyls)-thiazol-2-yl]-preparation of 3-hydroxyl-7-morpholine-4-base-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D00711
Be used to from the material of embodiment 2.3 and embodiment 12.8 and adopt the program of embodiment 13.1 to 13.2,2-[5-(3,4-two chloro-benzyls)-thiazol-2-yl is provided]-3-hydroxyl-7-morpholine-4-base-pyrido [1,2-α] pyrimidin-4-one.
1H?NMR(300MHz，DMSO-d 6)δ3.14-3.21(m，4H)，3.74-3.82(m，4H)，4.33(s，2H)，7.36(dd，J＝8.2Hz，2.1Hz，1H)，7.53(d，J＝10.0Hz，1H)，7.62(d，J＝8.2Hz，1H)，7.67(d，J＝2.1Hz，1H)，7.74-7.86(m，1H)，7.96(s，1H)，8.01-8.06(m，1H)，11.18-11.28(brs，1H)
MS(ESI -)m/z487(M-1)
HPLC Method 797.1%/19.7 minute
Embodiment 13.7:2-[5-(4-fluoro-benzyl)-thiazol-2-yl]-preparation of 3-hydroxyl-7-morpholine-4-ylmethyl-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D00721
Step 1:
Will from the product of embodiment 2.1 (3.66g, 15.6mmol), TERT-BUTYL DIMETHYL CHLORO SILANE (3.52g) and imidazoles (2.66g) join among methylene dichloride/DMF (30mL/10mL) and at room temperature stirred the mixture 2 hours.With methylene dichloride (30mL) diluted mixture thing and wash organic phase with water, drying is filtered also and is concentrated under vacuum.Residue carries out column chromatography (hexane/ethyl acetate 4: 1) so that desired compounds (5.02g, 92%) to be provided.
1H?NMR(300MHz，CDCl 3)：δ0.32(s，6H)，0.99(s，9H)，2.39(s，3H)，3.97(s，3H)，7.42(dd，J＝9.1，1.8Hz，1H)，7.62(d，J＝9.2Hz，1H)，8.68(bs，1H)。
Step 2:
Under nitrogen atmosphere, (5g adds N-bromosuccinimide (4.1g) and tert-butyl peroxide (0.348g) in stirred solution 14mmol) to the product in tetracol phenixin (80mL) from step 1.Reaction mixture refluxed 5 hours is cooled to room temperature then.With methylene dichloride (200mL) diluting soln, wash with water, drying is filtered and is concentrated under vacuum.Residue is carried out column chromatography (hexane/ethyl acetate 8: 1) so that the desired compounds as yellow solid (3.0g, 48%) to be provided.
1H?NMR(300MHz，DMSO-d 6)：δ0.26(s，6H)，0.94(s，9H)，3.86(s，3H)，4.88(s，2H)，7.66(d，J＝9.2Hz，1H)，7.79(dd，J＝9.3，2.0Hz，1H)，9.03(d，J＝1.8Hz，1H)
Step 3:
Will from the product of step 2 (1.1g, 2.6mmol) and morpholine (672mg, 7.73mmol) be dissolved in methylene chloride (1: 1, in mixed solvent 20mL).At room temperature stirred solution is 4 hours, partly concentrates under vacuum then and with methylene dichloride (40mL) dilution, it is washed with salt solution, and drying is filtered also vapourisation under reduced pressure.The purifying that is undertaken by silica gel column chromatography (hexane/ethyl acetate 1: 1) provides the product (1.03g, 92%) of expectation.
1H?NMR(300MHz，DMSO-d 6)：δ0.26(s，6H)，0.93(s，9H)，2.43(t，J＝4.5Hz，4H)，3.53-3.62(m，6H)，3.86(s，3H)，7.64(dd，J＝9.1，0.6Hz，1H)，7.76(dd，J＝9.2，1.9Hz，1H)，8.74(dd，J＝1.8，0.6Hz，1H)
Step 4:
Will (100mg, (1: 1: 3, mixing in the solvent and at room temperature stirring the mixture 5mL) be spent the night 0.23mmol) to join Glacial acetic acid/water/tetrahydrofuran (THF) from the product of step 4.Add water (10mL), adding solid sodium bicarbonate then is about 7 to regulate pH.With the organic layer that twice in dichloromethane extraction mixture and washing merge, dry and concentrated under vacuum, to provide desired compounds (65mg, 88%).
1H?NMR(300MHz，DMSO-d 6)：δ2.42(t，J＝4.5Hz，4H)，3.53-3.63(m，6H)，3.88(s，3H)，7.58(d，J＝9.2Hz，1H)，7.64(dd，J＝9.4，1.7Hz，1H)，8.62-8.67(m，1H)，10.24(s，1H)
Step 5-9:
The program that employing is described in embodiment 8.1 (except using DMF to react as solvent down at 70 ℃), embodiment 8.2, embodiment 12.2, embodiment 13.1 and embodiment 12.4 prepares 2-[5-(4-fluoro-benzyl)-thiazol-2-yl]-3-hydroxyl-7-morpholine-4-ylmethyl-pyrido [1,2-α] pyrimidin-4-one.
1H NMR (300MHz, DMSO-d 6) δ 2.40 (m, 4H, N-CH 2-CH 2-O), 3.53 (s, 2H, Ar-CH2-N), 3.57 (t, J=4.7Hz, 4H, N-CH 2-CH 2-O), 4.30 (s, 2H, CH 2-thiazole), 7.17 (t, J=8.9Hz, 2H, ArH), 7.39 (dd, J=8.9Hz, 5.4Hz, 2H, ArH), 7.52 (d, J=8.9Hz, 1H, H9), 7.65 (dd, J=8.9,2.4Hz, 1H, H8), (7.95 s, 1H, CH (thiazole)), 8.66 (m, 1H, H6), 11.33 (s, 1H, OH).
MS(ESI +)m/z453(M+1)
Embodiment 14: the 3-hydroxyl-4-oxo-4 of replacement, the preparation of 10-dihydro-benzo [4,5] imidazo [l, 2-α] pyrimidine-2-carboxylic acid benzyl acid amides: general route
Figure G2007800515627D00741
Come the amino benzoglyoxaline of alkylation 2-by adopting the program of in WO2005/058869, describing.The program that employing is described in embodiment 5 prepares methyl esters, and by adopt the program of describing in embodiment 6, it is converted to amide derivatives.Come purifying final product (formate is provided) by recrystallize or preparation HPLC.By adopting said procedure to prepare following examples (14.1-14.17).
Embodiment 14.1:3-hydroxyl-4-oxo-4,10-dihydro-benzo [4,5] imidazo [l, 2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00751
1H?NMR(300MHz，D6DSMO)：δ9.38(1H，m，NHCH 2)，8.43(1H，d，J＝8.1Hz，Ar-CH)，7.72-7.25(6H，m，Ar-CH)，4.52(2H，d，J＝6.3Hz，NHCH 2)。
MS(ESI -)m/z403(M-1)
Embodiment 14.2:3-hydroxyl-4-oxo-4, the preparation of 10-dihydro-benzo [4,5] imidazo [l, 2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
1H?NMR(300MHz，D6?DMSO)：δ9.27(1H，m，NHCH 2)，8.44(1H，d，J＝8.1Hz，Ar-CH)，7.50-7.14(7H，m，Ar-CH)，4.51(2H，d，J＝6.3Hz，NHCH 2)。
MS(ESI +)m/z353(M+1)。
Embodiment 14.3:3-hydroxyl-4-oxo-4,10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two fluoro-benzyl acid amides
Figure G2007800515627D00761
1H?NMR(300MHz，D6-DMSO)：δ9.31(1H，t，J＝6.3Hz，NHCH 2)，8.44(1H，d，J＝8.1Hz，Ar-CH)，7.51-7.18(6H，m，Ar-CH)，4.51(2H，d，J＝6.3Hz，NHCH 2)。
MS(ESI -)m/z370(M-1)。
Embodiment 14.4:3-hydroxyl-10-methyl-4-oxo-4, the preparation of 10-dihydro-benzo [4,5] imidazo [l, 2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D00762
1H?NMR(300MHz，D6?DMSO)：δ11.89(OH)，9.61(1H，t，J＝6.6Hz，NHCH 2)，8.44(1H，d，J＝7.8Hz，Ar-CH)，7.54(2H，m，Ar-CH)，7.41(2H，dd，J＝9.0，5.7Hz，Ar-CH)，7.35-7.30(1H，m，Ar-CH)，7.17(2H，m，Ar-CH)，4.54(2H，d，J＝6.6Hz，NHCH 2)，3.78(3H，s，CH 3)。
MS(ESI +)m/z367(M+1)。
Embodiment 14.5:3-hydroxyl-10-methyl-4-oxo-4, the preparation of 10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D00763
1H?NMR(300MHz，D6DMSO)：δ11.71(OH)，9.61(1H，t，J＝6.6Hz，NHCH 2)，8.44(1H?d，J＝8.1Hz，Ar-CH)，7.63-7.52(4H，m，Ar-CH)，7.38-7.34(2H，m，Ar-CH)，4.54(2H，d，J＝6.6Hz，NHCH 2)，3.79(3H，s，CH 3)。
MS(ESI +)m/z417(M+1)。
Embodiment 14.6:10-(4-fluoro-benzyl)-3-hydroxyl-4-oxo-4,10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00771
1H?NMR(300MHz，D6DMSO)：δ11.8(1H，s，OH)，9.75(1H，bs，NH)，8.47(1H，d，J＝4.8Hz，Ar-CH)，7.64(1H，d，J＝5.1Hz，Ar-CH)，7.63(1H，s，Ar-CH)，7.55(1H，d，J＝5.1Hz，Ar-CH)，7.54(1H，d，J＝4.8Hz，Ar-CH)，7.48(1H，dd，J＝4.8，4.5Hz，Ar-CH)，7.45(1H，dd，J＝4.8，4.2Hz，Ar-CH)，7.38-7.32(2H，m，Ar-CH)，7.17(1H，d，J＝5.4Hz，Ar-CH)，7.15(1H，d，J＝5.1Hz，Ar-CH)，5.61(2H，s，Ar-CH 2)，4.57(2H，d，J＝3.6Hz，CH 2NH)。
MS(ESI +)m/z511(M+1)。
Embodiment 14.7:3-hydroxyl-10-(2-morpholine-4-base-ethyl)-4-oxo-4,10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00772
1H?NMR(300MHz，D6DMSO)：δ11.77(1H，s，OH)，9.60(1H，t，J＝6.4Hz，NH)，8.46(1H，d，J＝8.4Hz，Ar-CH)，7.67-7.51(4H，m，Ar-CH)，7.35(2H，d，J＝8.1Hz，Ar-CH)，4.53(2H，d，J＝6.4Hz，NHCH 2)，4.49(2H，t，J＝6.0Hz，NCH 2CH 2NH)，3.30(4H，m，CH 2OCH 2)，2.69(2H，t，J＝6.0Hz，NCH 2CH 2N)，2.94-2.43(4H，m，CH 2NCH 2)。
MS(ESI +)m/z516(M) +。
Embodiment 14.8:3-hydroxyl-4-oxo-10-(2-tetramethyleneimine-l-base-ethyl)-4,10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00781
1H NMR (300MHz, D6DMSO): δ 8.09 (1H, s, NH), 7.70-7.65 (3H, m, Ar-CH), 7.37-7.29 (4H, m, Ar-CH), 5.30 (0.7H, s, tautomer B NCH 2), 4.62 (1.3H, d, J=6.3Hz, tautomer A NCH 2), 4.31 (2H, t, J=6.9Hz, NCH 2CH 2N), 2.98-2.92 (2H, m, NCH 2CH 2N), 2.65 (2H, m, CH 2NCH 2), 2.59 (2H, m, CH 2NCH 2), 1.82-1.77 (4H, m, NCH 2CH 2CH 2).
MS(ESI +)m/z500(M) +。
Embodiment 14.9:3-hydroxyl-4-oxo-10-(2-piperidines-l-base-ethyl)-4,10-dihydro-benzo [4,5] imidazo [l, 2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00782
1H?NMR(300MHz，D6DMSO)：δ11.58(1H，bs，OH)，8.69(1H，d，J＝7.8Hz，Ar-CH)，8.12(1H，bs，NH)，7.48(2H，d，J＝7.0Hz，Ar-CH)，7.45(1H，d，J＝8.4Hz，Ar-CH)，7.35-7.30(2H，m，Ar-CH)，7.23(1H，dd，J＝8.1，1.8Hz，Ar-CH)，4.62(2H，d，J＝6.0Hz，CH 2NH)，4.31(2H，t，J＝6.9Hz，NCH 2CH 2N)，2.72(2H，t，J＝6.9Hz，NCH 2CH 2N)，2.46(4H，t，J＝5.4Hz，CH 2NCH 2)，1.62-1.41(6H，m，NCH 2CH 2CH 2CH 2)。
MS (ESI +) m/z514 and 516 (M) +
Embodiment 14.10:3-hydroxyl-10-(2-methoxyl group-ethyl)-4-oxo-4,10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00791
1H?NMR(300MHz，D6DMSO)：δ11.78(1H，s，OH)，9.66(1H，bs，NH)，8.45(1H，d，J＝8.1Hz，Ar-CH)，7.65-7.60(3H，m，Ar-CH)，7.53(1H，m，Ar-CH)，7.37-7.30(2H，m，Ar-CH)，4.55(2H，d，J＝6.3Hz，CH 2NH)，4.52(2H，t，J＝5.4Hz，OCH 2CH 2N)，3.72(2H，t，J＝5.4Hz，OCH 2CH 2N)，3.27(3H，s，OCH 3)。
MS (ESI +) m/z461 and 463 (M+1).
Embodiment 14.11:3-hydroxyl-4-oxo-10-(3-piperidines-l-base-propyl group)-4,10-dihydro-benzo [4,5] imidazo [l, 2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00792
1H NMR (300MHz, D6DMSO): δ 11.70 (1H, bs, OH), 9.39 (1H, t, J=6.3Hz, NHCH 2), 8.47 (1H, d, J=7.8Hz, Ar-CH), 7.65-7.51 (4H, m, Ar-CH), 7.37-7.24 (2H, m, Ar-CH), 4.42 (2H, t, J=6.0Hz, CH 2N), 4.31 (2H, d, J=6.3Hz, NHCH 2), 2.25 (2H, t, J=6.0Hz, CH 2N), 1.99-1.92 (6H, m, CH 2NCH 2And NCH 2CH 2CH 2N), 1.17-1.11 (6H, m, NCH 2CH 2CH 2CH 2).
MS (ESI +) m/z528 and 530 (M+1).
Embodiment 14.12:3-hydroxyl-7,8-dimethyl-4-oxo-4,10-dihydro-benzo [4,5] imidazo [l, 2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00801
1H?NMR(300MHz，D6DMSO)：δ9.30(1H，t，J＝6.6Hz，NH)，8.22(1H，s，Ar-CH)，7.60(2H，m，Ar-CH)，7.35(1H，d，J＝8.1Hz，Ar-CH)，7.21(1H，s，Ar-CH)，4.51(2H，d，J＝6.6Hz，CH 2NH)，2.34(6H，s，2×CH 3)。
MS(ESI +)m/z413(M) +。
Embodiment 14.13:3-hydroxyl-7,8-dimethyl-10-(2-morpholine-4-base-ethyl)-4-oxo-4,10-dihydro-benzo [4,5] imidazo [l, 2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00802
1H?NMR(300MHz，D6DMSO)：δ11.70(1H，s，OH)，9.53(1H，bs，NH)，8.25(1H，s，Ar-CH)，7.62(1H，d，J＝8.1Hz，Ar-CH)，7.59(1H，d，J＝2.4Hz，Ar-CH)，7.44(1H，s，Ar-CH)，7.33(1H，dd，J＝8.1，2.4Hz，Ar-CH)，4.53(2H，d，J＝6.3Hz，NHCH 2)，4.44(2H，t，J＝5.7Hz，NCH 2CH 2N)，3.31(4H，m，CH 2OCH 2)，2.68(2H，t，J＝5.7Hz，NCH 2CH 2N)，2.43(4H，m，CH 2NCH 2)，2.38(3H，s，CH 3)，2.35(3H，s，CH 3)。
MS (ESI +) m/z544 and 546 (M+1).
Embodiment 14.14:3-hydroxyl-10-(2-morpholine-4-base-ethyl)-4-oxo-4, the preparation of 10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine-2-carboxylic acid 3-chloro-4-fluoro-benzyl acid amides
Figure G2007800515627D00811
1H?NMR(300MHz，CDCl 3)δ2.52(4H，bdd，J＝4.5Hz，2×-NCH 2CH 2O-)，2.79(2H，t，J＝6.3，7.2Hz，-NCH 2 CH 2NAr-)，3.59(4H，bdd，J＝4.2Hz，2×-NCH 2CH 2O-)，4.31(t，J＝6.6Hz，-NCH 2 CH 2NAr-)，4.61(2H，dd，J＝6.0Hz，-NHCH 2-)，7.15(1H，t，J＝8.4，8.7Hz，ArCH)，7.25(1H，dd，J＝2.4，4.5Hz?ArCH)，7.32(2H，m，ArCH)，7.42(1H，dd，J＝2.4，6.9Hz?ArCH)，7.50(2H，dt，J＝1.5，7.8Hz，ArCH)，8.00(1H，s，NH)，8.72(1H，s，NH)，8.68(1H，d，J＝8.1Hz，ArCH)，11.62(1H，s，OH)。
MS(ESI +)m/z500(M[Cl 35]+1)
Embodiment 14.15:3-hydroxyl-10-(2-morpholine-4-base-ethyl)-4-oxo-4, the preparation of 10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine-2-carboxylic acid 4-chloro-3-trifluoromethyl-benzyl acid amides
Figure G2007800515627D00821
1H?NMR(300MHz，CDCl 3)δ2.51(4H，bdd，J＝4.5Hz，2×-NCH 2CH 2O-)，2.77(2H，t，J＝6.6Hz，-NCH 2 CH 2NAr-)，3.59(4H，bdd，J＝4.5Hz，2×-NCH 2CH 2O-)，4.31(t，J＝6.6Hz，-NCH 2 CH 2NAr-)，4.68(2H，dd，J＝6.0Hz，-NHCH 2-)，7.30(2H，m，ArCH)，7.47(2H，dt，J＝0.9，8.0Hz，ArCH)，7.50(2H，bdd，J＝0.9，ArCH)，7.68(1H，bs，ArCH)，8.07(1H，d，J＝7.8Hz，ArCH)，11.55(1H，s，OH)。
MS(ESI +)m/z550(M[Cl 35]+1)
Embodiment 14.16:3-hydroxyl-10-(2-morpholine-4-base-ethyl)-4-oxo-4, the preparation of 10-dihydro-benzo [4,5] imidazo [l, 2-α] pyrimidine-2-carboxylic acid (3,4-two chloro-benzyls)-methyl-acid amides (formate)
1H?NMR(300MHz，CDCl 3)δ2.52(4H，bss，J＝4.5Hz，2×-NCH 2CH 2O-)，2.79(2H，t，J＝6.3，7.2Hz，-NCH 2 CH 2NAr-)，3.59(4H，bdd，J＝4.2Hz，2×-NCH 2CH 2O-)，4.31(t，J＝6.6Hz，-NCH 2 CH 2NAr-)，4.61(2H，dd，J＝6.0Hz，-NHCH 2-)，7.15(1H，t，J＝8.4，8.7Hz，ArCH)，7.25(1H，dd，J＝2.4，4.5Hz?ArCH)，7.32(2H，m，ArCH)，7.42(1H，dd，J＝2.4，6.9Hz?ArCH)，7.50(2H，dt，J＝1.5，7.8Hz，ArCH)，8.00(1H，s，NH)，8.72(1H，s，NH)，8.68(1H，d，J＝8.1Hz，ArCH)，11.62(1H，s，OH)。
MS (ESI +) m/z530 (M[Cl 35]+1-salt)
Embodiment 14.17:2-[2-(3,4-two chloro-phenyl)-tetramethyleneimine-l-carbonyl]-preparation of 3-hydroxyl-10-(2-morpholine-4-base-ethyl)-10H-benzo [4,5] imidazo [l, 2-α] pyrimidin-4-one
1H?NMR(300MHz，CDCl 3)δ2.52(4H，bdd，J＝4.5Hz，2×-NCH 2CH 2O-)，2.79(2H，t，J＝6.3，7.2Hz，-NCH 2 CH 2NAr-)，3.59(4H，bdd，J＝4.2Hz，2×-NCH 2CH 2O-)，4.31(t，J＝6.6Hz，-NCH 2 CH 2NAr-)，4.61(2H，dd，J＝6.0Hz，-NHCH 2-)，7.15(1H，t，J＝8.4，8.7Hz，ArCH)，7.25(1H，dd，J＝2.4，4.5Hz?ArCH)，7.32(2H，m，ArCH)，7.42(1H，dd，J＝2.4，6.9Hz?ArCH)，7.50(2H，dt，J＝1.5，7.8Hz，ArCH)，8.00(1H，s，NH)，8.72(1H，s，NH)，8.68(1H，d，J＝8.1Hz，ArCH)，11.62(1H，s，OH)
MS(ESI +)m/z556(M[Cl 35]+1)
Embodiment 15:3-hydroxyl-4-oxo-10-(2-piperazine-l-base-ethyl)-4, the preparation of 10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine-2-carboxylic acid benzyl acid amides: general method
Figure G2007800515627D00832
The preparation of embodiment 15.1.1:4-(2-chloro-ethyl)-piperazine-l-carboxylic acid tert-butyl ester
Figure G2007800515627D00841
Prepared 4-(2-chloro-ethyl)-piperazine-1-carboxylic acid tert-butyl ester according to the patented procedure of in WO2002/44141, describing.
1H?NMR(300MHz，CDCl 3)：δ3.45(2H，t，J＝6.9Hz，NCH 2)，3.40(4H，t，J＝4.8Hz，CH 2NCH 2)，2.52(2H，t，J＝6.9Hz，CH 2Cl)，2.42(4H，t，J＝4.8Hz，CH 2NCH 2)，1.49(9H，s，C[CH 3] 3。
MS(ESI +)m/z249(M+1)。
Embodiment 15.1.2:4-[2-(2-amino-benzoglyoxaline-l-yl)-ethyl]-preparation of piperazine-1-carboxylic acid tert-butyl ester
Figure G2007800515627D00842
Prepared 4-[2-(2-amino-benzoglyoxaline-1-yl)-ethyl according to the program of in WO2005/058869, describing]-piperazine-1-carboxylic acid tert-butyl ester.
1H?NMR(300MHz，CDCl 3)：δ7.43(1H，d，J＝7.5Hz，Ar-CH)，7.17-7.06(3H，m，Ar-CH)，5.78(2H，s，NH 2)，3.64(2H，t，J＝5.7Hz，NCH 2CH 2N)，3.46(4H，m，CH 2NCH 2)，2.56(4H，m，CH 2NCH 2)，2.43(2H，t，J＝5.7Hz，NCH 2CH 2N)，1.46(9H，s，C[CH 3] 3)。
MS(ESI +)m/z346(M+1)。
Embodiment 15.2:3-acetoxyl group-10-[2-(4-tert-butoxycarbonyl-piperazine-l-yl)-ethyl]-4-oxo-4, the preparation of 10-dihydro-benzo [4,5] imidazo [l, 2-α] pyrimidine-2-carboxylate methyl ester
Figure G2007800515627D00851
By adopting the program shown in the embodiment 5 to prepare 3-acetoxyl group-10-[2-(4-tert-butoxycarbonyl-piperazine-1-yl)-ethyl]-4-oxo-4,10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine-2-carboxylate methyl ester.
1H?NMR(300MHz，D6DMSO)：δ8.66(1H，d，J＝8.7Hz，Ar-CH)，7.58(1H，m，Ar-CH)，7.42(2H，m，Ar-CH)，4.29(2H，t，J＝5.9Hz，NCH 2CH 2N)，3.96(3H，s，OCH 3)，3.55(4H，t，J＝5.4Hz，CH 2NCH 2)，2.88(2H，t，J＝5.9Hz，NCH 2CH 2N)，2.70(4H，t，J＝5.4Hz，CH 2NCH 2)，2.20(3H，s，O＝CCH 3)，1.47(9H，s，C[CH 3] 3)。
MS(ESI +)m/z514(M+1)。
Embodiment 15.3:3-acetoxyl group-4-oxo-10-(2-piperazine-l-base-ethyl)-4, the preparation of 10-dihydro-benzo [4,5] imidazo [l, 2-α] pyrimidine-2-carboxylate methyl ester
Figure G2007800515627D00852
Handle 3-acetoxyl group-10-[2-(4-tert-butoxycarbonyl-piperazine-1-yl)-ethyl with trifluoroacetic acid (0.20mL)]-4-oxo-4,10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine-2-carboxylate methyl ester (33mg, 0.064mmol), and at room temperature stirred 1 hour.After this, enriched mixture and thick residue are the product (27mg, 100%) of expectation.
1H NMR (300MHz, D6-DMSO): δ 8.76 (1H, bs, NH), 8.47 (1H, d, J=8.1Hz, Ar-CH), 7.85 (1H, d, J=8.1Hz, Ar-CH), 7.65 (1H, dd, J=8.1,7.8Hz, Ar-CH), 7.47 (1H, dd, J=8.1,7.8Hz, Ar-CH), 4.57 (2H, t, J=5.4Hz, NCH 2CH 2N), 3.88 (3H, s, OCH 3), 3.27-3.07 (10H, m, 2 * NCH 2CH 2N and 8 * NHCH 2CH 2NCH 2CH 2), 2.31 (3H, s, [C=O] CH 3).
MS(ESI +)m/z414(M+1)。
Embodiment 15.4:4-{2-[2-(3,4-two chloro-benzylamino formyl radicals)-3-hydroxyl-4-oxo-4H-benzo [4,5] imidazo [l, 2-α] pyrimidine-10-yl]-ethyl }-preparation of piperazine-l-carboxylic acid tert-butyl ester
Figure G2007800515627D00861
According to by embodiment 6 improved programs; to change into 4-{2-[2-(3 from the product of embodiment 15.2; 4-two chloro-benzylamino formyl radicals)-3-hydroxyl-4-oxo-4H-benzo [4,5] imidazo [1,2-α] pyrimidine-10-yl]-ethyl }-piperazine-1-carboxylic acid tert-butyl ester.
1H?NMR(300MHz，D6DMSO)：δ8.68(1H，d，J＝7.5Hz，Ar-CH)，8.02(1H，t，J＝6.3Hz，NHCH 2)，7.49-7.14(6H，m，Ar-CH)，4.63(2H，d，J＝6.3Hz，NHCH 2)，4.31(2H，t，J＝6.9Hz，NCH 2CH 2N)，3.34(4H，m，CH 2NCH 2)，2.79(2H，t，J＝6.9Hz，NCH 2CH 2N)，2.45(4H，m，CH 2NCH 2)，1.47(9H，s，C[CH 3] 3)。
MS (ESI +) m/z615 and 617 (M+1).
Embodiment 15.5:3-hydroxyl-4-oxo-10-(2-piperazine-l-base-ethyl)-4,10-dihydro-benzo [4,5] imidazo [l, 2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00871
According to by embodiment 6 improved programs, will change into 3-hydroxyl-4-oxo-10-(2-piperazine-1-base-ethyl)-4 from the product of embodiment 15.3,10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine-2-carboxylic acid 3,4-two chloro-benzyl acid amides.
1H?NMR(300MHz，D6DMSO)：δ9.65(1H，t，J＝6.6Hz，NHCH 2)，8.46(1H，d，J＝8.1Hz，Ar-CH)，7.68-7.60(3H，m，Ar-CH)，7.58-7.50(1H，m，Ar-CH)，7.38-7.31(2H，m，Ar-CH)，4.54(2H，d，J＝6.6Hz，NHCH 2)，4.47(2H，t，J＝6.0Hz，NCH 2CH 2N)，3.16(4H，m，CH 2NCH 2)，2.66(2H，t，J＝6.0Hz，NCH 2CH 2N)，2.41(4H，m，CH 2NCH 2)。
MS (ESI +) m/z 515 and 517 (M+1).
Embodiment 15.6:3-hydroxyl-10-[2-(4-methylsulfonyl-piperazine-l-yl)-ethyl]-4-oxo-4,10-dihydro-benzo [4,5] imidazo [l, 2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00872
With 3-acetoxyl group-4-oxo-10-(2-piperazine-1-base-ethyl)-4,10-dihydro-benzo [4,5] imidazo [1,2-a] pyrimidine-2-carboxylate methyl ester (embodiment 15.3) (97mg, 0.235mmol) be dissolved in the methylene dichloride (1mL), and to wherein add triethylamine (98 μ L, 0.71mmol), then add methylsulfonyl chloride (21 μ L, 0.26mol).Stirring reaction at room temperature; make solvent evaporation then and residue changed into 3-hydroxyl-10-[2-(4-methylsulfonyl-piperazine-1-yl)-ethyl by the program that adopts description among the embodiment 6]-4-oxo-4; 10-dihydro-benzo [4; 5] imidazo [1; 2-α] pyrimidine-2-carboxylic acid 3,4-two chloro-benzyl acid amides.
1H?NMR(300MHz，D6DMSO)：δ9.60(1H，t，J＝6.3Hz，NHCH 2)，8.47(1H，d，J＝8.1Hz，Ar-CH)，7.68-7.52(4H，m，Ar-CH)，7.36-7.31(2H，m，Ar-CH)，4.55(2H，d，J＝6.3Hz，NHCH 2)，4.51(2H，t，J＝5.7Hz，NCH 2CH 2N)，2.87(4H，m，CH 2NCH 2)，2.79(2H，t，J＝5.7Hz，NCH 2CH 2N)，2.74(3H，s，SCH 3)，2.57(4H，m，CH 2NCH 2)。
MS (ESI +) m/z593 and 595 (M+1).
Embodiment 15.7:10-[2-(4-ethanoyl-piperazine-l-yl)-ethyl]-3-hydroxyl-4-oxygen-4,10-dihydro-benzo [4,5] imidazo [l, 2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00881
With 3-acetoxyl group-4-oxo-10-(2-piperazine-1-base-ethyl)-4,10-dihydro-benzo [4,5] imidazo [1,2-a] pyrimidine-2-carboxylate methyl ester (embodiment 15.3) (100mg, 0.242mmol) be dissolved in the methylene dichloride (1mL), and to wherein add triethylamine (96 μ L, 0.70mmol), then add Acetyl Chloride 98Min. (21 μ L, 0.266mol).At room temperature stirring reaction is 3 hours; make solvent evaporation then; and by adopting the program of describing among the embodiment 6 that residue is changed into 10-[2-(4-ethanoyl-piperazine-1-yl)-ethyl]-3-hydroxyl-4-oxo-4; 10-dihydro-benzo [4; 5] imidazo [1; 2-a] pyrimidine-2-carboxylic acid 3,4-two chloro-benzyl acid amides.
1H?NMR(300MHz，D6DMSO)：δ9.61(1H，t，J＝6.3Hz，NHCH 2)，8.46(1H，d，J＝8.4Hz，Ar-CH)，7.70-7.62(3H，m，Ar-CH)，7.57(1H，m，Ar-CH)，7.37(2H，m，Ar-CH)，4.56(2H，d，J＝6.3Hz，NHCH 2)，4.51(2H，t，J＝5.7Hz，NCH 2CH 2N)，3.20(2H，m，CH 2NCH 2)，3.12(2H，m，CH 2NCH 2)，2.73(2H，t，J＝5.7Hz，NCH 2CH 2N)，2.45-2.38(4H，m，CH 2NCH 2)，1.89(3H，s，CH 3)。
MS(ESI +)m/z?557(M) +。
Embodiment 15.8:3-hydroxyl-10-[2-(4-methyl-piperazine-l-yl)-ethyl]-4-oxygen-4,10-dihydro-benzo [4,5] imidazo [l, 2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00891
With 3-acetoxyl group-4-oxo-10-(2-piperazine-1-base-ethyl)-4,10-dihydro-benzo [4,5] imidazo [1,2-a] pyrimidine-2-carboxylate methyl ester (embodiment 15.3) (97mg 0.235mmol) is dissolved in the methyl alcohol (1mL), and to wherein adding sodium cyanoborohydride (21mg, 0.63mmol) and sodium acetate (30mg, 0.38mmol), then add formaldehyde (38 μ L, 0.47mmol).At room temperature stirring reaction is 2 hours, make solvent evaporation then and residue changed into 3-hydroxyl-10-[2-(4-methyl-piperazine-1-yl)-ethyl by the program that adopts description among the embodiment 6]-4-oxo-4,10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine-2-carboxylic acid 3,4-two chloro-benzyl acid amides.
1H?NMR(300MHz，D6DMSO)：δ9.58(1H，t，J＝6.3Hz，NHCH 2)，8.46(1H，d，J＝7.2Hz，Ar-CH)，7.66-7.53(4H，m，Ar-CH)，7.36-7.31(2H，m，Ar-CH)，4.54(2H，d，J＝6.3Hz，NHCH 2)，4.47(2H，t，J＝6.0Hz，NCH 2CH 2N)，2.67(2H，t，J＝6.0Hz，NCH 2CH 2N)，2.54-2.26(8H，m，CH 2NCH 2CH 2NCH 2)，1.99(3H，s，CH 3)。
MS(ESI +)m/z?527(M) +。
Embodiment 15.9.1:3-acetoxyl group-10-[2-(4-methoxymethyl-piperazine-l-yl)-ethyl]-4-oxo-4, the preparation of 10-dihydro-benzo [4,5] imidazo [l, 2-α] pyrimidine-2-carboxylate methyl ester
With 3-acetoxyl group-4-oxo-10-(2-piperazine-1-base-ethyl)-4,10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine-2-carboxylate methyl ester (embodiment 15.3) (97mg, 0.235mmol) be dissolved in the methylene dichloride (1mL), and to wherein add diisopropylethylamine (95 μ L, 0.52mmol), then add the methoxymethyl muriate (20 μ L, 0.258mmol).At room temperature stirring reaction is 24 hours, make then solvent evaporation and by column chromatography (98: 1.5: 0.5 methylene dichloride: methyl alcohol: purifying residue ammoniacal liquor), with the product (30mg, 28%) that expectation is provided.
1H?NMR(300MHz，D6-DMSO)：δ8.68(1H，d，J＝8.4HzAr-CH)，7.54(1H，dd，J＝8.1，7.8Hz，Ar-CH)，7.41-7.35(2H，m，Ar-CH)，4.42(2H，t，J＝6.3Hz，NCH 2CH 2N)，3.99(3H，s，OCH 3)，3.60(2H，s，CH 2OCH 3)，3.52-3.45(2H，m，CH 2N)，3.31(2H，m，CH 2N)，2.83(2H，t，J＝6.3Hz，NCH 2CH 2N)，2.55-2.46(4H，m，CH 2NCH 2)，2.09(3H，s，OCH 3)，2.06(3H，s，O＝CCH 3)。
MS(ESI +)m/z458(M+1)。
Embodiment 15.9.2:3-hydroxyl-10-[2-(4-methoxymethyl-piperazine-l-yl)-ethyl]-4-oxo-4, l0-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Utilization is by embodiment 6 improved programs, to change into 3-hydroxyl-10-[2-(4-methoxymethyl-piperazine-1-yl)-ethyl from the product of embodiment 15.9.1]-4-oxo-4,10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidine-2-carboxylic acid 3,4-two chloro-benzyl acid amides.
1H?NMR(300MHz，D6?DMSO)：δ8.70(1H，d，J＝8.1Hz，Ar-CH)，7.90(1H，m，NH)，7.52-7.19(6H，m，Ar-CH)，4.63(2H，d，J＝6.0Hz，CH 2NH)，4.32(2H，t，J＝6.6Hz，NCH 2CH 2N)，3.85(2H，s，CH 2OCH 3)，3.65(2H，m，CH 2N)，3.50(2H，m，CH 2N)，2.81(2H，t，J＝6.6Hz，NCH 2CH 2N)，2.51(4H，m，CH 2NCH 2)，2.04(3H，s，OCH 3)。
MS (ESI +) m/z557 and 559 (M+1).
Embodiment 15.10: methylsulfonic acid 2-(3,4-two chloro-benzylamino formyl radicals)-10-[2-(4-methylsulfonyl-piperazine-l-yl)-ethyl]-4-oxo-4, the preparation of 10-dihydro-benzo [4,5] imidazo [1,2-α] pyrimidin-3-yl ester
With 3-hydroxyl-4-oxo-10-(2-piperazine-1-base-ethyl)-4,10-dihydro-benzo [4,5] imidazo [1,2-a] pyrimidine-2-carboxylic acid 3,4-two chloro-benzyl acid amides (embodiment 15.5) (75mg, 0.146mmol) and triethylamine (60 μ L 0.31mmol) are dissolved in the methylene dichloride (1mL), and to wherein add methylsulfonyl chloride (18 μ L, 0.31mmol).At room temperature stirring reaction is 15 minutes, thereafter under vacuum concentrated solvent and by column chromatography (95: 4.5: 0.5 methylene dichloride: methyl alcohol: purifying residue ammoniacal liquor), with the product (65mg, 76%) that expectation is provided.
1H?NMR(300MHz，D6?DMSO)：δ8.48(1H，t，J＝6.6Hz，NHCH 2)，7.82(1H，d，J＝8.1Hz，Ar-CH)，7.62(4H，m，Ar-CH)，7.35(2H，m，Ar-CH)，4.57(2H，m，CH 2N)，4.49(2H，t，J＝6.0Hz，NCH 2CH 2N)，4.16(2H，d，J＝6.6Hz，NHCH 2)，2.88(4H，m，CH 2NCH 2)，2.78(2H，m，NCH 2CH 2N)，2.55(2H，m，CH 2N)，2.06(6H，s，2×S-CH 3)。
MS (ESI +) m/z671 and 673 (M+1).
Embodiment 16: the 6-hydroxyl of replacement-5-oxo-5H-thiazole is the preparation of [3,2-α] pyrimidine-7-carboxylate methyl ester also
Embodiment 16.1:6-(2,2-dimethyl-propionyloxy)-5-oxo-5H-thiazole is the preparation of [3,2-α] pyrimidine-7-carboxylate methyl ester also
Step 1:
The program that to describe in embodiment 5 is used for thiazolamine so that the ester of expectation to be provided.
1H?NMR(300MHz，DMSO-d 6)δ3.86(s，3H)，7.55(d，J＝4.7Hz，1H))7.96(d，J＝4.9Hz，1H)，10.21(s，1H)
Step 2:
With above-mentioned ester (620mg, 2.7mmol) and triethylamine (2.21g 21mmol) is dissolved in the methylene dichloride (30mL).At room temperature in above-mentioned solution, drip trimethyl-acetyl chloride (362mg, 3.0mmol).After finishing adding, stirred the mixture 1 hour, under vacuum, concentrate then.Come the product (610mg, 83%) of purifying residue by the column chromatography that utilizes (hexane/ethyl acetate 1: 1) so that expectation to be provided.
1H?NMR(300MHz，DMSO-d 6)δ1.30(s，9H)，3.84(s，3H)，7.73(d，J＝5.0Hz，1H)，8.08(d，J＝4.8Hz，1H)
MS(ESI +)m/z?333(M+Na +)
Embodiment 16.2:6-hydroxy-2-methyl-5-oxo-5H-thiazole is the preparation of [3,2-α] pyrimidine-7-carboxylate methyl ester also
Figure G2007800515627D00922
The program that to describe in embodiment 5 is used for 2-amino-5-methylthiazol so that the ester of expectation to be provided.
1H?NMR(300MHz，DMSO-d 6)δ2.41(d，J＝1.4Hz，3H)，3.85(s，3H)，7.81(d，J＝1.5Hz，1H)，10.21(s，1H)。
MS(ESI +)m/z?263(M+Na +)
Embodiment 16.3:6-hydroxy-3-methyl-5-oxo-5H-thiazole is the preparation of [3,2-α] pyrimidine-7-carboxylate methyl ester also
By handling with diacetyl oxide 2-amino-4-methylthiazol is changed into the N-acetic ester under 90 ℃, productive rate is 80%.With mCPBA it is carried out oxidation, productive rate is 45%, and hydrolysis is to provide the N-oxide compound of 2-amino-4-methylthiazol then, and it stands the Chem. at J.Heterocyclic, and the condition described in 1979,16 is to provide thick ester.The program that utilization is described in embodiment 16.1 (step 2) is carried out acidylate so that the product of expectation to be provided to this crude ester.
1H?NMR(300MHz，CDCl 3)δ1.41(s，9H)，2.82(s，3H)，3.93(s，3H)，6.55(s，1H)
Embodiment 16.4:6-hydroxyl-2-sec.-propyl-5-oxo-5H-thiazole is the preparation of [3,2-α] pyrimidine-7-carboxylate methyl ester also
Figure G2007800515627D00932
The program that to describe in embodiment 5 is used for 2-amino-5-sec.-propyl thiazole so that the ester of expectation to be provided.
1H?NMR(300MHz，DMSO-d 6)δ1.29(d，J＝7.0Hz，6H)，3.12-3.23(m，1H)，3.85(s，3H)，7.75(d，J＝1.2Hz，1H)，10.23(s，1H)。
MS(ESI -)m/z267(M-1)
Embodiment 17: the 6-hydroxyl of replacement-5-oxo-5H-thiazole is the preparation of [3,2-α] pyrimidine-7-carboxylic acid benzyl acid amides also
Embodiment 17.1:6-hydroxyl-5-oxo-5H-thiazole is the preparation of [3,2-α] pyrimidine-7-carboxylic acid 4-fluoro-benzyl acid amides also
Figure G2007800515627D00941
Begin and adopt the program of embodiment 6 by product, obtained also [3,2-α] pyrimidine-7-carboxylic acid 4-fluoro-benzyl acid amides of 6-hydroxyl-5-oxo-5H-thiazole from embodiment 16.1.
1H?NMR(300MHz，D6-DMSO)：δ12.35(1H，s，OH)，9.74(1H，t，J＝6.3Hz，CH 2NH)，7.97(1H，d，J＝5.1Hz，Ar-CH)，7.52(1H，d，J＝5.1Hz，Ar-CH)，7.38(2H，dd，J＝7.8，8.0Hz，Ar-CH)，7.58(2H，dd，J＝7.8，8.0Hz，Ar-CH)，4.45(2H，d，J＝6.3Hz，CH 2NH)。
MS(ESI -)m/z318(M-H)。
Embodiment 17.2:6-hydroxyl-5-oxo-5H-thiazole is [3,2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl acid amides
Begin and adopt the program of embodiment 6 by product, obtained also [3,2-α] pyrimidine-7-carboxylic acid 3 of 6-hydroxyl-5-oxo-5H-thiazole, 4-two chloro-benzyl acid amides from embodiment 16.1.
Figure G2007800515627D00942
1H?NMR(300MHz，D6-DMSO)：δ12.22(1H，s，OH)，9.79(1H，t，J＝5.7Hz，CH 2NH)，7.97(1H，d，J＝5.4Hz，Ar-CH)，7.60(1H，d，J＝8.1Hz，Ar-CH)，7.60(1H，s，Ar-CH)，7.52(1H，d，J＝5.4Hz，Ar-CH)，7.34(1H，d，J＝8.1Hz，Ar-CH)，4.47(2H，d，J＝5.7Hz，CH 2NH)。
MS(ESI -)m/z368(M[Cl 35]-1)
HPLC Method 799.1%/15.4 minute
Embodiment 17.3:6-hydroxy-2-methyl-5-oxo-5H-thiazole is the preparation of [3,2-α] pyrimidine-7-carboxylic acid 4-fluoro-benzyl acid amides also
Figure G2007800515627D00951
Begin and adopt the program of embodiment 6 by product, obtained also [3,2-α] pyrimidine-7-carboxylic acid 4-fluoro-benzyl acid amides of 6-hydroxy-2-methyl-5-oxo-5H-thiazole from embodiment 16.2.
1H?NMR(300MHz，D6-DMSO)δ2.38(3H，s，-CHC(S)CH 3)，4.42(2H，d，J＝5.7Hz，-NH-CH 2-)，7.13(2H，m，ArH)，7.37(2H，m，ArH)，7.79(1H，s，-CHC(S)CH 3)，9.72(1H，t，J＝6.3Hz，-NHCH 2-)。
MS(ESI -)m/z332(M[Cl 35]-1)。
HPLC Method 798.4%/10.6 minute
Embodiment 17.4:6-hydroxy-2-methyl-5-oxo-5H-thiazole is [3,2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00952
Begin and adopt the program of embodiment 6 by product, obtained also [3,2-α] pyrimidine-7-carboxylic acid 3 of 6-hydroxy-2-methyl-5-oxo-5H-thiazole, 4-two chloro-benzyl acid amides from embodiment 16.2.
1H?NMR(300MHz，D6-DMSO)：δ12.21(1H，s，OH)，9.76(1H，t，J＝6.3Hz，CH 2NH)，7.81(1H，s，Ar-CH)，7.59(2H，d，J＝7.8Hz，Ar-CH)，7.33(1H，d，J＝7.8Hz，Ar-CH)，4.46(2H，d，J＝6.3Hz，CH 2NH)，2.40(3H，s，CH 3)。
MS(ESI -)m/z382(M[Cl 35]-1)。
HPLC Method 799.1%/14.6 minute
Embodiment 17.5:6-hydroxy-3-methyl-5-oxo-5H-thiazole is [3,2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00961
Begin and adopt the program of embodiment 6 by product, obtained also [3,2-α] pyrimidine-7-carboxylic acid 3 of 6-hydroxy-3-methyl-5-oxo-5H-thiazole, 4-two chloro-benzyl acid amides from embodiment 16.3.
1H?NMR(300MHz，CDCl 3)δ2.80(3H，s，-(CH)C(N)CH 3)，4.57(2H，d，J＝6.6Hz，-NH-CH 2-)，6.37(1H，s，-(CH 3)C＝CH-S-)，7.18(1H，m，ArH)，7.43(2H，m，ArH)，7.95(1H，m，-NHCH 2-)。
MS(ESI -)m/z332(M[Cl 35]-1)。
HPLC Method 799.6%/10.6 minute
Embodiment 17.6:6-hydroxyl-2-sec.-propyl-5-oxo-5H-thiazole is [3,2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D00962
Begin and adopt the program of embodiment 6 by product, obtained also [3,2-α] pyrimidine-7-carboxylic acid 3 of 6-hydroxyl-2-sec.-propyl-5-oxo-5H-thiazole, 4-two chloro-benzyl acid amides from embodiment 16.4.
1H?NMR(300MHz，D6-DMSO)δ1.27(6H，d，2×(CH 3) 2CH-)，3.15(1H，m，CH 3) 2CH-)，4.44(2H，d，J＝6.6Hz，-(O＝C)NHCH 2-)，7.31(1H，dd，J＝1.8，8.1Hz，ArH)，7.58(2H，m，ArH)，7.74(1H，d，J＝1.2Hz，ArH)，9.78(1H，t，J＝6.0Hz，-(O＝C)NHCH 2-)，12.22(1H，s，OH)。
MS(ESI +)m.z434(M[Cl 35]+Na)。
HPLC Method 799.0%/15.2 minute
Embodiment 17.7: the 2-amino methyl of replacement-6-hydroxyl-5-oxo-5H-thiazole is the preparation-general method of [3,2-α] pyrimidine-7-carboxylic acid benzyl acid amides also
Figure G2007800515627D00971
Step 1
Figure G2007800515627D00972
Cooling (ice/water-bath) is from the product in methylene dichloride (50mL) (660mg, stirred solution 2.75mmol) of embodiment 16.2.Add N, and the N-dimethyl aminopyridine (500mg, 4.13mmol), and after 10 minutes, dripping acetyl chloride (320mg, 4.13mmol).With mixture heating up to room temperature and stirred 6 hours.TLC shows that initial ester is consumed, and with aqueous hydrochloric acid (4.0M), salt solution purging compound twice, dry (Na 2SO 4), filter and under vacuum, concentrate, to provide desired compounds (70mg, 90%) as yellow solid.
1H?NMR(300MHz，CDCl 3)δ2.38(s，3H)，2.49(s，3H)，3.96(s，3H)，7.70(s，1H)
MS(ESI +)m/z?305(M+23)
Step 2
Figure G2007800515627D00981
(6g 21mmol), the N-bromosuccinimide (3.02g) in tetracol phenixin (400mL) and tert-butyl peroxide (1.03g), and is heated to backflow from the intermediate of step 1 in merging.After 1 hour, add the N-bromosuccinimide (1.14g) of another part.Make reaction mixture refluxed other 4 hours, and be cooled to room temperature then.By solid collected by filtration, be dissolved in the methylene dichloride then, and by water washing solution, drying also is evaporated to drying, so that crude product to be provided.Carry out further purifying by recrystallize from dichloromethane/hexane the desired compounds as white solid (3.0g, 39%) is provided.
1H?NMR(300MHz，DMSO-d 6)δ2.31(s，3H)，3.87(s，3H)，5.02(d，J＝0.9Hz，2H)，8.33(t，J＝0.8Hz，1H)。
MS(ESI +)m/z?383(M[Br 79]+23)，385(M[Br 81]+23)
Step 3:A
At room temperature be stirred in the product (0.33mmol) of the step 2 in the methylene dichloride (6mL) and amine (1mmol) 20 hours.Collect resulting throw out by filtering,, be directly used in the next step reaction then with the cold methanol washing.
Step 3:B
At room temperature, in stirred solution, drip amine (2.49mmol) from the product (0.83mmol) in methylene dichloride (20mL) of step 3:A.Mixture was kept at room temperature 24 hours.(1.0M 20mL) extracts with methylene dichloride (20mL) diluted mixture thing and with aqueous hydrochloric acid.Utilize aqueous NaOH (1.0M) that water is adjusted to pH=10, (2 * 30mL) extract to use methylene dichloride then.Dry (Na 2SO 4) organic layer that merges, filter also and under vacuum, concentrate.Be further purified product by recrystallize or by preparation HPLC.
Step 4:A
The program that employing is described in embodiment 6.Be further purified product by recrystallize or by preparation HPLC.
Step 4:B
In from the stirred solution of the product (0.14mmol) in methyl alcohol (20mL) of step 3:B, add 1.3 normal benzylamines.The mixture of backflow gained 24 hours concentrates under vacuum then.Residue is dissolved in the methylene dichloride (20mL) and with aqueous NaOH (1.0M, 10mL) washing, in organic layer, add then aqueous hydrochloric acid (1.0M, 15mL).By filter collecting the throw out of gained, with cold water, then use hexane wash.Drying solid is to provide the desired compounds as hydrochloride under vacuum.
Embodiment 17.7.1:6-hydroxyl-2-morpholine-4-ylmethyl-5-oxo-5H-thiazole is the preparation of [3,2-α] pyrimidine-7-carboxylic acid 4-fluoro-benzyl acid amides also
Utilize step 3:A and step 4:A to prepare following compound:
Figure G2007800515627D00991
1H?NMR(300MHz，D6-DMSO)δ2.43(4H，bs，N-CH 2-CH 2-O)，3.56(4H，bs，N-CH 2-CH 2-O)，4.43(2H，d，J＝6.3Hz，-NH-CH 2-)，7.14(2H，m，ArH)，7.36(2H，m，ArH)，7.96(1H，s，S-C＝CH-N-)，9.74(1H，bs，O＝C-NH-CH 2)。
MS(ESI -)m/z?419(M[Cl 35]-1)
HPLC Method 796.7%/12.2 minute
Embodiment 17.7.2:6-hydroxyl-2-morpholine-4-ylmethyl-5-oxo-5H-thiazole is [3,2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl acid amides
Utilize step 3:A and step 4:A to prepare following compound:
Figure G2007800515627D01001
1H?NMR(300MHz，D6-DMSO)δ2.49(4H，s，N-CH 2-CH 2-O)，3.61(4H，s，N-CH 2-CH 2-O)，4.49(2H，d，J＝6.3Hz，-NH-CH 2-)，7.36(1H，dd，J＝8.4，2.1Hz，ArH)，7.62(2H，m，ArH)，8.03(1H，s，S-C＝CH-N-)，9.82(1H，bt，O＝C-NH-CH 2)，12.25(1H，s，OH)。
MS(ESI -)m/z467(M[Cl 35]-1)
HPLC Method 799.1%/13.9 minute
Embodiment 17.7.3:6-hydroxyl-5-oxo-2-piperidines-l-ylmethyl-5H-thiazole is [3,2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl acid amides
Utilize step 3:A and step 4:A to prepare following compound:
Figure G2007800515627D01002
1H?NMR(300MHz，D6-DMSO)：δ11.84(1H，s，OH)，8.03(1H，m，NHCH 2)，7.77(1H，s，Ar-CH)，7.45(2H，m，Ar-CH)，7.18(1H，d，J＝8.7Hz，Ar-CH)，4.58(2H，d，J＝6.3Hz，NHCH 2)，3.56(2H，s，NCH 2[C])，2.45(4H，m，CH 2NCH 2)，1.59-1.45(6H，m，NCH 2CH 2CH 2CH 2)。
MS(ESI -)m/z465(M[Cl 35]-1)
HPLC Method 797.7%/10.3 minute
Embodiment 17.7.4: the preparation of dibutyl-[7-(3,4-two chloro-benzylamino formyl radicals)-6-hydroxyl-5-oxo-5H-thiazole is [3,2-α] pyrimidine-2-base methyl also]-ammonium (muriate)
Utilize step 3:B and step 4:B to prepare following compound:
Figure G2007800515627D01011
1H?NMR(300MHz，D6-DMSO)：δ12.25(1H，s，OH)，10.79(1H，bs，N +HCl)，9.80(1H，t，J＝6.0Hz，NHCH 2).8.30(1H，s，Ar-CH)，7.57(1H，d，J＝6.6Hz，Ar-CH)，4.56(2H，bs，NCH 2[C])，4.45(2H，d，J＝6.0Hz，CH 2NH)，3.01(4H，m，2×NCH 2CH 2)，1.66(4H，m，2×NCH 2CH 2CH 2)，1.31(4H，m，2×NCH 2CH 2CH 2CH 3)，0.89(6H，t，J＝7.2Hz，2×NCH 2CH 2CH 2CH 3)。
MS(ESI -)m/z509(M[Cl 35]-1)
HPLC Method 798.5%/11.5 minute
Embodiment 17.7.5:6-hydroxyl-5-oxo-2-piperazine-l-ylmethyl-5H-thiazole is [3,2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl acid amides
Utilize step 3:A and step 4:A to prepare following compound:
Figure G2007800515627D01012
1H?NMR(300MHz，D6-DMSO)：δ10.94(1H，s，OH)，7.85(1H，s，Ar-CH)，7.59(1H，d，J＝8.4Hz，Ar-CH)，7.55(1H，s，Ar-CH)，7.30(1H，d，J＝8.4Hz，Ar-CH)，4.47(2H，d，J＝6.0Hz，CH 2NH)，4.32(2H，s，NCH 2[C])，2.89(4H，m，CH 2NCH 2)，2.50(4H，m，CH 2NCH 2)。
MS(ESI -)m/z466(M[Cl 35]-1)
HPLC Method 796.3%/9.97 minute
Embodiment 17.7.6:6-hydroxyl-2-(4-methyl-piperazine-l-ylmethyl)-5-oxo-5H-thiazole is [3,2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl acid amides
Utilize step 3:A and step 4:A to prepare following compound:
Figure G2007800515627D01021
1H?NMR(300MHz，DMSO)δ2.17(3H，-N-CH 3)，2.33(4H，bs，N-CH 2-CH 2-O)，2.46(4H，bs，N-CH 2-CH 2-O)，4.44(2H，d，J＝6.3Hz，-NH-CH 2-)，7.30(1H，dd，J＝2.1，8.1Hz，ArH)，7.57(2H，m，ArH)，7.94(1H，s，S-C＝CH-N-)，9.87(1H，bt，O＝C-NH-CH 2)。
MS(ESI +)m/z482(M[Cl 35]+1)
HPLC Method 795.9%/10.3 minute
Embodiment 17.7.7:6-hydroxyl-2-(3-methyl-piperazine-l-ylmethyl)-5-oxo-5H-thiazole is [3,2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl acid amides
Utilize step 3:A and step 4:A to prepare following compound:
Figure G2007800515627D01022
1H?NMR(300MHz，D6-DMSO)：δ7.82(1H，s，Ar-CH)，7.57(1H，d，J＝8.1Hz，Ar-CH)，7.54(1H，s，Ar-CH)，7.29(1H，d，J＝8.1Hz，Ar-CH)，4.56(2H，d，J＝5.7Hz，CH 2NH)，3.69(2H，s，CH 2N)，3.67-2.83(5H，m，NCH[CH 3]CH 2NCH 2)，2.14(1H，t，J＝6.3Hz，CH 2N)，1.86(1H，t，J＝6.3Hz，CH 2N)，1.02(3H，d，J＝6.3Hz，CH 3CH)。
MS(ESI -)m/z480(M[Cl 35]-1)
HPLC Method 781.4%/10.1 minute
Embodiment 17.7.8:2-diethylamino methyl-6-hydroxyl-5-oxo-5H-thiazole is [3,2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl amide hydrochlorides
Utilize step 3:B and step 4:B to prepare following compound:
Figure G2007800515627D01031
1H?NMR(300MHz，D6-DMSO)：δ12.25(1H，s，OH)，11.09(1H，bs，NH +Et 2)，9.81(1H，t，J＝6.3Hz，NHCH 2)，8.32(1H，s，Ar-CH)，7.59(1H，d，J＝8.1Hz，Ar-CH)，7.56(1H，s，Ar-CH)，7.32(1H，d，J＝8.1Hz，Ar-CH)，4.54(2H，s，CH 2NH +)，4.46(2H，d，J＝6.3Hz，NHCH 2)，3.06(4H，m，2×CH 2CH 3)，1.26(6H，m，2×CH 2CH 3)。
MS(ESI -)m/z453(M[Cl 35]-1)
HPLC Method 798.5%/10.2 minute
Embodiment 17.7.9:6-hydroxyl-5-oxo-2-tetramethyleneimine-l-ylmethyl-5H-thiazole is [3,2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl amide hydrochlorides
1H?NMR(300MHz，D6-DMSO)：δ12.26(1H，s，OH)，11.01(1H，bs，NH +)，9.80(1H，t，J＝6.3Hz，CH 2NH)，8.26(1H，s，Ar-CH)，7.58(1H，d，J＝8.2Hz，Ar-CH)，7.57(1H，s，Ar-CH)，7.31(1H，d，J＝8.1Hz，Ar-CH)，4.57(2H，bs，CH 2NH +)，4.45(2H，d，J＝6.3Hz，CH 2NH)，3.47(2H，m，CH 2N)，3.10(2H，m，CH 2N)，2.04-1.85(4H，m，NCH 2CH 2CH 2)。
MS(ESI -)m/z451(M[Cl 35]-1)
HPLC Method 799.4%/10.1 minute
Embodiment 17.7.10:2-dimethylaminomethyl-6-hydroxyl-5-oxo-5H-thiazole is [3,2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl amide hydrochlorides
Utilize step 3:B and step 4:B to prepare following compound:
Figure G2007800515627D01041
1H?NMR(300MHz?D6-DMSO)：δ12.26(1H，s，OH)，11.18(1H，bs，NH +)，9.81(1H，t，J＝6.3Hz，CH 2NH)，8.25(1H，s，Ar-CH)，7.60(1H，d，J＝8.1Hz，Ar-CH)，7.58(1H，s，Ar-CH)，7.33(1H，d，J＝8.1Hz，Ar-CH)，4.51(2H，bs，CH 2NH +)，4.48(2H，d，J＝6.3Hz，CH 2NH)，2.75(6H，s，2×CH 3)。
MS(ESI -)m/z427(M[Cl 35]-1)
HPLC Method 798.0%/9.9 minute
Embodiment 17.7.11:4-[7-(3,4-two chloro-benzylamino formyl radicals)-6-hydroxyl-5-oxo-5H-thiazole is [3,2-α] pyrimidine-2-base methyl also]-preparation of piperazine-l-carboxylic acid tert-butyl ester
Utilize step 3:A and step 4:A to prepare following compound:
Figure G2007800515627D01042
1H?NM[R(300MHz，D6-DMSO)：δ12.24(1H，s，OH)，9.80(1H，m，CH 2NH)，8.00(1H，s，Ar-CH)，7.61(1H，d，J＝8.4Hz，Ar-CH)，7.59(1H，s，Ar-CH)，7.33(1H，d，J＝8.4Hz，Ar-CH)，4.46(2H，d，J＝6.0Hz，CH 2NH)，3.71(2H，s，CH 2N)，3.33(4H，m，CH 2NCH 2)，2.41(4H，m，CH 2NCH 2)，1.40(9H，s，C[CH 3] 3)。
MS(ESI -)m/z566(M[Cl 35]-1)
HPLC Method 798.3%/11.4 minute
Embodiment 17.7.12:6-hydroxyl-2-[4-(2-methoxyl group-phenyl)-piperazine-l-ylmethyl]-5-oxo-5H-thiazole [3,2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl acid amides
Utilize step 3:A and step 4:A to prepare following compound:
Figure G2007800515627D01051
1H?NMR(300MHz，D6-DMSO)δ2.61(4H，bs，2×-NCH 2CH 2N-CH 2-)，2.96(4H，bs，2×-NCH 2CH 2N-CH 2-)，3.60(2H，s，-NCH 2CH 2N-CH 2-)，3.75(3H，s，-OCH 3)，4.45(2H，dd，J＝6.6Hz，-(O＝C)NHCH 2-)，6.90(4H，m，ArH)，7.26(1H，dd，J＝1.5，9.1Hz，ArH)，7.53(3H，m，ArH)，12.1(1H，bs，OH)。
MS(ESI -)m/z572(M[Cl 35]-1)
HPLC Method 797.0%/11.2 minute
Embodiment 17.7.13:6-hydroxyl-5-oxo-2-propyl group amino methyl-5H-thiazole is [3,2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl amide hydrochlorides
Utilize step 3:B and step 4:B to prepare following compound:
Figure G2007800515627D01052
1H?NMR(300MHz，D6-DMSO)δ0.91(3H，t，J＝7.2Hz，CH 3CH 2-)，1.65(2H，q，J＝7.2Hz，CH 3CH 2-)，2.87(2H，t，J＝7.2Hz，-CH 2CH 2-)，4.37(2H，s，-CH 2CH 2NH 2 +CH 2-)，4.47(2H，dd，J＝6.6Hz，-(O＝C)NHCH 2-)，7.33(1H，dd，J＝2.1，8.4Hz，ArH)，7.59(2H，m，ArH)，8.23(1H，s，ArH)，9.36(1H，bs，-(O＝C)NHCH 2-)，9.79(1H，t，J＝6.0Hz，ArH)，12.3(1H，bs，OH).
MS(ESI -)m/z439(M[Cl 35]-1)
HPLC Method 796.0%/10.2 minute
Embodiment 17.7.14:6-hydroxyl-5-oxo-2-(4-phenyl-Piperazine-l-ylmethyl)-5H-thiazole is [3,2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl acid amides
Utilize step 3:A and step 4:A to prepare following compound:
Figure G2007800515627D01061
1H?NMR(300MHz，D6-DMSO)δ2.62(4H，bs，PhNCH 2CH 2N-)，3.14(4H，bs，PhNCH 2CH 2O-)，3.74(2H，s，PhNCH 2CH 2NCH 2-)，4.46(2H，d，J＝6.0Hz，-(O＝C)NHCH 2-)，6.77(2H，t，J＝7.5Hz，ArH)，6.93(2H，d，J＝8.1Hz，ArH)，7.20(2H，t，J＝7.5Hz，ArH)，7.32(1H，dd，J＝1.8，8.2Hz，ArH)，7.60(2H，m，ArH)，8.0(1H，s，ArH)9.88(1H，t，J＝6.6Hz，-(O＝C)NHCH 2-)。
MS(ESI -)m/z542(M[Cl 35]-1)
HPLC Method 775.0%/11.4 minute
Embodiment 17.7.15:6-hydroxyl-2-(4-methylsulfonyl-piperazine-l-ylmethyl)-5-oxo-5H-thiazole is [3,2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl acid amides
Utilize step 3:A and step 4:A to prepare following compound:
Figure G2007800515627D01062
1H?NMR(300MHz，D6-DMSO)δ2.54(4H，bs，(SO 2CH 3)NCH 2CH 2N-)，2.88(3H，bs，(SO 2CH 3)NCH 2CH 2N-)，3.14(4H，bs，(SO 2CH 3)NCH 2CH 2N-)，3.65(2H，s，-NCH 2CH 2NCH 2-)，4.43(2H，bs，-(O＝C)NHCH 2-)，7.25(1H，bs，ArH)，7.51(2H，bs，ArH)，7.71(1H，bs，ArH)。
MS(ESI -)m/z544(M[Cl 35]-1)
HPLC Method 792.0%/10.9 minute
Embodiment 18: the 6-hydroxyl-5-oxo-l of replacement, 5-dihydro-imidazol-be the preparation of [l, 2-a] pyrimidine-7-carboxylic acid benzyl acid amides also
Embodiment 18.1 general methods
Figure G2007800515627D01071
Embodiment 18.1.1: route A
Embodiment 18.1.1.1:6-hydroxyl-5-oxo-l, 5-dihydro-imidazol-be the preparation (step 1) of [l, 2-α] pyrimidine-7-carboxylate methyl ester also
Figure G2007800515627D01072
(7.4g 56mmol) is suspended in the anhydrous methanol (20mL) and is cooled to-78 ℃ with 2-aminooimidazole Monosulfate.In this mixture, slowly be added in sodium methylate (3.0g, solution 56mmol) in the anhydrous methanol (20mL).After adding is finished, mixture heating up to room temperature was also kept at room temperature 4 hours.By solid collected by filtration and with the anhydrous methanol washing, merge washing lotion then and under vacuum, be concentrated into drying, so that 2-aminooimidazole (4.1g, productive rate 90%) to be provided.
1H?NMR(300MHz，DMSO-d 6)δ4.95-5.30(brs，2H)，6.40(s，2H)
In the 25mL flask, mix the diacetoxyl dimethyl fumarate (3.2g, 12mmol), the 2-aminooimidazole (1.03g, 12mmol) and tosic acid (395mg, 2.0mmol) and be immersed in the pre-hot oil bath (120 ℃).After 6 hours, reaction mixture is cooled to room temperature and ethyl acetate (10mL) and methyl alcohol (0.6mL) are joined in the residue, the sonication mixture is 2 minutes then.By filtering the throw out of collecting gained, use cold ethyl acetate (1mL) washing and use the pump drying, to provide crude product.To provide 6-hydroxyl-5-oxo-1, the 5-dihydro-imidazol-is [1,2-α] pyrimidine-7-carboxylate methyl ester (1.03g, 41%) also from the methyl alcohol recrystallize.
1H?NMR(300MHz，DMSO-d 6)δ3.85(s，3H)，7.56-7.64(m，2H)，9.00-9.40(brs，1H)，12.30-12.70(brs，1H)
Embodiment 18.1.1.2:6-acetoxyl group-5-oxo-l, 5-dihydro-imidazol-be the preparation (step 2) of [l, 2-a] pyrimidine-7-carboxylate methyl ester also:
Will (300mg, 1.43mmol) and N, (262mg 2.15mmol) be dissolved in the methylene dichloride (20mL) the N-dimethyl aminopyridine, is cooled to 5 ℃ then from the product of embodiment 18.1.1.1.In stirring the mixture, this drips Acetyl Chloride 98Min. (112mg, solution 1.43mmol) in methylene dichloride (5mL).Remaining under this temperature after 30 minutes, with mixture heating up to room temperature and stirred other 12 hours.
Under reduced pressure remove and desolvate, and utilize methylene chloride (20: 1) to make resulting residue stand silica gel column chromatography as elutriant.Acquisition is as the 6-acetoxyl group-5-oxo-1 of yellow solid, and the 5-dihydro-imidazol-is [1,2-α] pyrimidine-7-carboxylate methyl ester (195mg, 54.2%) also.
1H?NMR(300MHz，DMSO-d 6)δ2.26(s，3H)，3.84(s，3H)，7.72(d，J＝2.6Hz，1H)，7.75(d，J＝2.6Hz，1H)，13.20-13.26(brs，1H)
Embodiment 18.1.1.3:6-acetoxyl group-l-(2-morpholine-4-base-ethyl)-5-oxo-l, 5-dihydro-imidazol-be the preparation (step 3) of [l, 2-α] pyrimidine-7-carboxylate methyl ester also
Figure G2007800515627D01091
At room temperature, make product from embodiment 18.1.1.2 (30mg, 0.12mmol) and hexaoxacyclooctadecane-6-6 (3mg 10%w/w) mixes with acetonitrile (5mL).In this stirred solution, add Anhydrous potassium carbonate (83mg, 0.60mmol) and 4-(2-chloroethyl)-morpholine hydrochloride (25mg, 0.132mmol).Heated mixt is 2 hours under refluxing, and is cooled to room temperature then.Concentration response is near dry and add ethyl acetate (15mL), water (2 * 10mL) purging compounds then under vacuum.Separate organic layer, dryly also under vacuum, concentrate, then by utilizing methylene chloride (20: 1) to come the purifying residue as the column chromatography of elutriant.Acquisition is as 6-acetoxyl group-1-(2-morpholine-4-base-ethyl)-5-oxo-1 of white solid, and the 5-dihydro-imidazol-is [1,2-α] pyrimidine-7-carboxylate methyl ester (15mg, 34%) also.
Embodiment 18.1.1.4:6-acetoxyl group-l-allyl group-5-oxo-l, 5-dihydro-imidazol-be the preparation of [l, 2-α] pyrimidine-7-carboxylate methyl ester also
The program that employing is described in embodiment 18.1.1.3, difference be, before adding allyl bromide 98 (1.1 equivalent) and reaction mixture is heated to 70 ℃ 30 minutes, at room temperature sodium hydride (1.1 equivalent) joined in the reaction mixture.
1H?NMR(500MHz，CDCl 3)：δ7.65(1H，d，J＝2.7Hz，Ar-CH)，7.10(1H，d，J＝2.7Hz，Ar-CH)，5.98(1H，ddt，J＝17.1，10.5，5.7Hz，CH 2CH＝CH 2)，5.39(2H，m，CH 2CH＝CH 2)，4.78(2H，dt，J＝5.7，1.8Hz，CH 2CH＝CH 2)，3.95(3H，s，OCH 3)，2.37(3H，s，C[＝O]CH 3)。
Embodiment 18.1.2: route B
Preparation (the step 1) of embodiment 18.1.2.1:l-(2-piperidines-l-base-ethyl)-1H-imidazoles-2-base amine
Figure G2007800515627D01101
With 2-aminooimidazole Monosulfate (500mg, 1.89mmol) and salt of wormwood (580mg 4.16mmol) is suspended among the DMF (2mL), and add chloroethyl piperidines monohydrochloride (766mg, 4.16mmol).100 ℃ of following reacting by heating 2.5 hours, be cooled to room temperature then and filter.Concentrated filtrate and by column chromatography (95: 4.5: 0.5 methylene dichloride: methyl alcohol: purifying in addition ammoniacal liquor), separate product (83mg, 11%) then as brown oil.
1H?NMR(300MHz，D6-DMSO)：δ6.59(1H，s，Ar-CH)，6.45(1H，s，Ar-CH)，5.41(2H，bs，NH 2)，3.80(2H，t，J＝5.0Hz，NCH 2CH 2N)，2.59(2H，t，J＝5.0Hz，NCH 2CH 2N)，2.46(4H，m，CH 2NCH 2)，1.57(4H，m，NCH 2CH 2CH 2CH 2)，1.45(2H，m，NCH 2CH 2CH 2CH 2)。
Embodiment 18.1.2.2:6-acetoxyl group-5-oxo-l-(2-piperidines-l-base-ethyl)-l, the 5-dihydro-imidazol-is the preparation (step 2) of [l, 2-a] pyrimidine-7-carboxylate methyl ester also
Figure G2007800515627D01102
The program that employing is described in embodiment 4 is to provide 6-acetoxyl group-5-oxo-1-(2-piperidines-1-base-ethyl)-1, and the 5-dihydro-imidazol-is [1,2-α] pyrimidine-7-carboxylate methyl ester also.
1H?NMR(300MHz，D6-DMSO)：δ7.60(1H，d，J＝2.4Hz，Ar-CH)，7.42(1H，d，J＝2.4Hz，Ar-CH)，4.33(2H，m，CH 2N)，3.95(3H，s，OCH 3)，2.76(2H，m，CH 2N)，2.54(4H，m，CH 2NCH 2)，2.37(3H，s，O＝CCH 3)，1.63-1.43(6H，m，NCH 2CH 2CH 2CH 2)。
MS(ESI +)m/z?363(M+1)。
Embodiment 18.2:6-hydroxyl-l-methyl-5-oxo-l, 5-dihydro-imidazol-be the preparation of [l, 2-α] pyrimidine-7-carboxylic acid 4-fluoro-benzyl acid amides also
Figure G2007800515627D01111
Begun and adopted the program of describing in embodiment 18.1.2 by methyl iodide, 6-hydroxyl-1-methyl-5-oxo-1 is provided, the 5-dihydro-imidazol-is [1,2-α] pyrimidine-7-carboxylate methyl ester also.By adopting the program of in embodiment 6, describing, 6-hydroxyl-1-methyl-5-oxo-1 is provided, the 5-dihydro-imidazol-is [1,2-α] pyrimidine-7-carboxylic acid 4-fluoro-benzyl acid amides also.
Embodiment 18.3:6-hydroxyl-l-methyl-5-oxo-l, 5-dihydro-imidazol-be [l, 2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D01112
Adopted the program of describing in embodiment 18.2, so that 6-hydroxyl-1-methyl-5-oxo-1 to be provided, the 5-dihydro-imidazol-is [1,2-α] pyrimidine-7-carboxylic acid 3 also, 4-two chloro-benzyl acid amides.
1H?NMR(300MHz，DMSO)δ3.67(3H，s，N-CH 3)，4.50(2H，d，J＝6.3Hz，-NH-CH 2-)，7.32(1H，dd，J＝2.1，8.1Hz，-(CH 3)N-CH-CH-N-)，7.58(4H，m，ArH)，9.56(1H，t，J＝6.3Hz，-NHCH 2-)。
MS(ESI -)m/z?365(M[Cl 35]-1)
Embodiment 18.4:6-hydroxyl-l-(2-morpholine-4-base-ethyl)-5-oxo-l, 5-dihydro-imidazol-be [l, 2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D01121
Utilize the product of embodiment 18.1.1.3 and by adopt the program of describing in embodiment 6, prepared 6-hydroxyl-1-(2-morpholine-4-base-ethyl)-5-oxo-1, the 5-dihydro-imidazol-is [1,2-α] pyrimidine-7-carboxylic acid 3 also, 4-two chloro-benzyl acid amides.
1H?NM[R(300MHz，D6-DMSO)：δ9.57(1H，t，J＝6.6Hz，NHCH 2)，7.67(1H，d，J＝2.7Hz，Ar-CH)，7.60(3H，m，Ar-CH)，7.33(1H，d，J＝8.4Hz，Ar-CH)，4.53(2H，d，J＝6.6Hz，NHCH 2)，7.26(2H，t，J＝5.4Hz，NCH 2CH 2N)，3.43(4H，t，J＝4.8Hz，CH 2OCH 2)，2.68(2H，t，J＝5.4Hz，NCH 2CH 2N)，2.45(4H，m，CH 2NCH 2)。
MS(ESI -)m/z464(M[Cl 35]-1)
Embodiment 18.5:6-hydroxyl-5-oxo-l-(2-piperidines-l-base-ethyl)-l, 5-dihydro-imidazol-be [l, 2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl acid amides
The program of embodiment 6 is used for product from embodiment 18.1.2.2 so that 6-hydroxyl-5-oxo-1-(2-piperidines-1-base-ethyl)-1 to be provided, and the 5-dihydro-imidazol-is [1,2-α] pyrimidine-7-carboxylic acid 3 also, 4-two chloro-benzyl acid amides.
1H?NMR(300MHz，D6-DMSO)：δ11.43(1H，s，OH)，9.55(1H，bs，NH)，7.64-7.59(4H，m，Ar-CH)，7.32(1H，d，J＝8.1Hz，Ar-CH)，4.51(2H，d，J＝6.6Hz，CH 2NH)，4.23(2H，t，J＝6.3Hz，NCH 2CH 2N)，2.62(2H，t，J＝6.3Hz，NCH 2CH 2NH)，2.41-2.38(4H，m，CH 2NCH 2)，1.23(6H，m，NCH 2CH 2CH 2CH 2)。
MS(ESI +)m/z464(M[Cl 35]+1)
Embodiment 18.6:l-allyl group-6-hydroxyl-5-oxo-l, 5-dihydro-imidazol-be [l, 2-α] pyrimidine-7-carboxylic acid 3 also, the preparation of 4-two chloro-benzyl acid amides
The program of describing in embodiment 6 is used for product from embodiment 18.1.1.4 so that the product of expectation to be provided.
1H?NMR(500MHz，D6-DMSO)：δ9.57(1H，m，NHCH 2)，7.66-7.57(4H，m，Ar-CH)，7.33(1H，d，J＝7.8Hz，Ar-CH)，6.10-5.97(1H，m，CH 2CH＝CH 2)，5.27(1H，m，CH 2CH＝CH 2)，5.23(1H，m，CH 2CH＝CH 2)，4.77(2H，d，J＝5.4Hz，CH 2CH＝CH 2)，4.51(2H，d，J＝6.6Hz，NHCH 2)。
MS(ESI +)m/z393(M[Cl 35]+1)。
Embodiment 19: the preparation of the 2-hydroxyl of replacement-l-oxo-1H-9-oxa--4,9 α-diaza-fluorenes-3-carboxylic acid benzyl acid amides
The preparation of embodiment 19.1:2-hydroxyl-l-oxo-1H-9-oxa--4,9 α-diaza-fluorenes-3-carboxylate methyl ester
Figure G2007800515627D01132
For step 1 and step 2, adopt at J.Heterocyclic Chem., the program of describing in 1989,26,1293 is to provide 3-amino-benzoisoxazole.The program that employing is described in embodiment 5 is to provide the ester of expectation.
1H?NMR(300MHz，D6-DMSO)δ3.90(s，3H)，7.52-7.62(m，1H)，7.80-7.90(m，2H)，8.12(d，J＝7.8Hz，1H)，10.92(s，1H)。
MS(ESI +)m/z?283(M+Na)。
The preparation of embodiment 19.2:2-hydroxyl-5-morpholine-4-base-l-oxo-lH-9-oxa--4,9 α-diaza-fluorenes-3-carboxylate methyl ester
By adopting the program of in embodiment 19.1, describing and utilizing 2-fluoro-6-morpholine-4-base-benzonitrile, prepared the ester of expectation as starting raw material.
1H?NMR(300MHz，D6-DMSO)δ3.24-3.45(4H)，3.85(t，J＝4.6Hz，4H)，3.90(s，3H)，6.89(d，J＝8.3Hz，1H)，7.25(d，J＝8.1Hz，1H)，7.68(t，J＝8.4Hz，1H)，10.78(s，1H)
The preparation of embodiment 19.3:2-benzyloxy-l-oxo-lH-9-oxa--4,9 α-diaza-fluorenes-3-carboxylate methyl ester
Figure G2007800515627D01142
The product from embodiment 19.1 of cooling (ice/water-bath) in tetrahydrofuran (THF) (10mL) (50mg, 0.19mmol) and phenylcarbinol (46mg, solution 0.42mmol).In this solution, add triphenylphosphine (111mg, 0.423mmol) and diisopropyl azodiformate (85mg, 0.42mmol).Mixture heating up to room temperature and after 2 hours, is concentrated volatile matter under vacuum.By column chromatography (hexane/ethyl acetate 1: 1) purifying residue so that the desired compounds as white solid (53mg, 79%) to be provided.
1H?NMR(300MHz，DMSO-d 6)δ3.82(s，3H)，5.23(s，2H)，7.31-7.52(m，5H)，7.57-7.67(m，1H)，7.93(d，J＝3.2Hz，2H)，8.18(d，J＝7.7Hz，1H)
The preparation of embodiment 19.4:2-benzyloxy-5-morpholine-4-base-l-oxo-lH-9-oxa--4,9 α-diaza-fluorenes-3-carboxylate methyl ester
Figure G2007800515627D01151
The program that employing is described in embodiment 8.1 wherein utilizes DMF to react as solvent under 70 ℃, so that desired compounds to be provided.
1H?NMR(300MHz，DMSO-d 6)δ3.33-3.40(m，4H)，3.77-3.86(m，7H)，5.21(s，2H)，6.93(d，J＝8.2Hz，1H)，7.29-7.50(m，6H)，7.75(t，J＝8.4Hz，1H)。
The preparation of embodiment 19.5:2-benzyloxy-l-oxo-1H-9-oxa--4,9 α-diaza-fluorenes-3-carboxylic acid
Figure G2007800515627D01152
Be used to obtain desired compounds from the product of embodiment 19.3 and the program of employing description in embodiment 8.2.
1H?NMR(300MHz，DMSO-d 6)δ5.21(s，2H)，7.30-7.45(m，3H)，7.50(d，J＝6.6Hz，2H)，7.56-7.69(m，1H)，7.93(d，J＝3.6Hz，2H)，8.19(d，J＝8.1Hz，1H)，13.78-13.98(brs，1H)。
The preparation of embodiment 19.6:2-benzyloxy-5-morpholine-4-base-l-oxo-1H-9-oxa--4,9 α-diaza-fluorenes-3-carboxylic acid
Figure G2007800515627D01153
Be used to from the product of embodiment 19.4 and adopt program described in the embodiment 8.2, obtained desired compounds.
1H?NMR(300MHz，DMSO-d 6)δ3.34-3.41(m，4H)，3.78-3.86(m，4H)，5.20(s，2H)，6.93(d，J＝8.2Hz，1H)，7.30-7.44(m，4H)，7.46-7.54(m，2H)，7.75(t，J＝8.2Hz，1H)，13.58-13.79(brs，1H)
MS(ESI -)m/z420(M-1)
Embodiment 19.7:2-benzyloxy-5-morpholine-4-base-l-oxo-lH-9-oxa--4,9a-diaza-fluorenes-3-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D01161
With N, N '-dicyclohexylcarbodiimide (110mg, 0.522mmol) (200mg is in stirred solution 0.475mmol), after 30 minutes to join the product from embodiment 19.6 in methylene dichloride (100mL), add N successively, the N-dimethyl aminopyridine (6mg, 0.05mmol), 3,4-dichloro-benzylamine (92mg, 0.52mmol) and I-hydroxybenzotriazole (70mg, 0.52mmol).At room temperature stir the mixture and spend the night, and water the crude product of assembling (aqueous work-up) and providing is provided is further purified as elutriant by column chromatography (hexane/ethyl acetate 4: 1), so that the product as the expectation of yellow solid (120mg, 44%) to be provided.
1H?NMR(300MHz，CDCl 3)δ3.37-3.45(m，4H)，3.85-3.95(m，4H)，4.51(d，J＝5.8Hz，2H)，5.43(s，2H)，6.84(d，J＝8.3Hz，1H)，7.06-7.16(m，2H)，7.28-7.36(m，3H)，7.37-7.49(m，4H)，7.55-7.71(m，2H)。
Embodiment 19.8:2-hydroxyl-5-morpholine-4-base-l-oxo-lH-9-oxa--4,9 α-diaza-fluorenes-3-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D01171
(10mg, (12mg is in stirred solution 0.021mmol) 0.062mmol) to join the product from embodiment 19.7 in methylene dichloride (5mL) with iron(ic) chloride (III).At room temperature stirred the mixture 1.5 hours, and dripped aqueous hydrochloric acid (1.0M) then and become transparent up to solution.With ethyl acetate extraction product and dry organic phase, under vacuum, concentrate then.Residue by mixed solvent (hexane/ethyl acetate 10/1) recrystallize so that the desired compounds as gray solid (8mg, 80%) to be provided.
1H?NMR(300MHz，D6-DMSO)：δ12.28(1H，s，OH)，8.62(1H，m，NHCH 2)，7.28(3H，m，Ar-CH)，7.40(1H，d，J＝8.7Hz，Ar-CH)，7.30(1H，d，J＝8.4Hz，Ar-CH)，6.95(1H，d，J＝8.1Hz，Ar-CH)，4.62(2H，d，J＝6.6Hz，NHCH 2)，3.77(4H，m，CH 2OCH 2)，2.49(4H，m，CH 2NCH 2)。
MS(ESI -)m/z487(M[Cl 35]-1)
Embodiment 19.9:2-hydroxyl-l-oxo-1H-9-oxa--4,9 α-diaza-fluorenes-3-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D01172
Being used to provides desired compounds from the product of embodiment 19.5 and the program of employing description in embodiment 19.7 and embodiment 19.8.
1H?NMR(300MHz，D6-DMSO)δ4.55(2H，d，J＝6.0Hz，-NH-CH 2-)，7.37(1H，dd，J＝8.4，2.1Hz，ArH)，7.61(3H，m，ArH)，7.87(2H，m，ArH)，8.07(1H，d，J＝7.5Hz，ArH)，9.77(1H，t，J＝6.0Hz，O＝C-NH-CH 2)，12.79(1H，s，OH)。
MS(ESI +)m/z404(M[Cl 35]+1)
HPLC Method 796.2%/19.0 minute
The preparation of embodiment 19.10:2-hydroxyl-l-oxo-1H-9-oxa--4,9 α-diaza-fluorenes-3-carboxylic acid 4-fluoro-benzyl acid amides
Being used to provides desired compounds from the product of embodiment 19.5 and the program of employing description in embodiment 19.7 and embodiment 19.8.
1H?NMR(300MHz，D6-DMSO)δ4.54(2H，d，J＝6.9Hz，-NH-CH 2-)，7.17(2H，t，J＝9.0，2.4Hz，ArH)，7.42(2H，m，ArH)，7.60(1H，m，ArH)，7.87(1H，m，ArH)，8.06(1H，d，J＝8.1Hz，ArH)，9.72(1H，t，J＝6.6Hz?O＝C-NH-CH 2)，12.93(1H，s，OH)。
MS(ESI -)m/z352(M-1)
HPLC Method 793.1%/12.5 minute
The preparation of embodiment 19.11:2-hydroxyl-5-morpholine-4-base-l-oxo-1H-9-oxa--4,9 α-diaza-fluorenes-3-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D01182
Being used to provides desired compounds from the product of embodiment 19.6 and the program of employing description in embodiment 19.7 and embodiment 19.8.
1H?NMR(300MHz，D6-DMSO)δ3.29(4H，s，N-CH 2-CH 2-O)，3.71(4H，s，N-CH 2-CH 2-O)，4.60(2H，dd，J＝6.0Hz，-NH-CH 2-)，6.94(1H，dd，J＝7.8Hz，ArH)，7.21(2H，t，J＝8.2Hz，ArH)，7.29(1H，dd，J＝8.4Hz，ArH)，7.45(2H，m，ArH)，7.70(1H，dd，J＝8.4Hz，ArH)，8.43(1H，t，O＝C-NH-CH 2)，12.42(1H，s，OH)。
MS(ESI -)m/z437(M[Cl 35]-1)
HPLC Method 791.0%/15.7 minute
Embodiment 20: the 3-hydroxyl of replacement-4-oxo-4H-9-thia-l, the preparation of 4 α-diaza-fluorenes-2-carboxylic acid benzyl acid amides
By adopting the program of in embodiment 4 and embodiment 6, describing, prepared following compound:
Embodiment 20.1:3-hydroxyl-4-oxo-4H-9-thia-l, 4 α-diaza-fluorenes-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D01191
1H?NMR(300MHz，D6-DMSO)：δ12.35(1H，s，OH)，9.82(1H，t，J＝6.9Hz，NHCH 2)，8.90(1H，m，Ar-CH)，8.01(1H，m，Ar-CH)，7.63-7.56(4H，m，Ar-CH)，7.35(1H，d，J＝7.8Hz，Ar-CH)，4.48(2H，d，J＝6.9Hz，CH 2NH)。
MS(ESI -)m/z418(M[Cl 35]-1)
HPLC Method 791%/18.8 minute
Embodiment 20.2:3-hydroxyl-7-methoxyl group-4-oxo-4H-9-thia-l, 4 α-diaza-fluorenes-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D01192
1H?NMR(300MHz，D6-DMSO)：δ12.30(1H，s，OH)，9.75(1H，t，J＝6.9Hz，NHCH 2)，8.77(1H，d，J＝9.3Hz，Ar-CH)，7.63-7.59(3H，m，Ar-CH)，7.33(1H，d，J＝8.4Hz，Ar-CH)，7.13(1H，d，J＝9.3Hz，Ar-CH)，4.46(2H，d，J＝6.9Hz，CH 2NH)，3.83(3H，s，CH 3)。
HPLC Method 795.3%/19.1 minute
Embodiment 21: the 3-hydroxyl-4-oxo-4 of replacement, the preparation of 6-dihydro-Mi Dingbing [1,2-b] indazole-2-carboxylic acid benzyl acid amides
Embodiment 21.1:3-hydroxyl-4-oxo-4, the preparation of 6-dihydro-Mi Dingbing [l, 2-b] indazole-2-carboxylate methyl ester
Figure G2007800515627D01201
Step 1:
(605mg, 5mmol) (352mg 6mmol) mixes with 1-butanols (3mL) with 85% moisture hydrazine hydrate with 2-fluorine benzonitrile.Heated mixt is 5 hours under backflow and stirring, is cooled to room temperature then.By filter to collect resulting throw out and use washed with dichloromethane, dry cake under vacuum then is to provide 3-amino benzopyrazoles (293mg, 44%).
1H?NMR(300MHz，D6-DMSO)：δ5.26-5.36(brs，2H)，6.84-6.93(m，1H)，7.18-7.24(m，2H)，7.67(dt，J＝8.1，0.9Hz，1H)，11.33(s，1H)。
Step 2:
The program of describing in embodiment 3 is used for product from step 1 so that desired compounds to be provided.
1H?NMR(300MHz，D6-DMSO)：δ3.91(s，3H)，7.33(t，J＝7.7Hz，1H)，7.48(dt，J＝8.0，0.8Hz，1H)，7.70(t，J＝7.7Hz，1H)，8.09(d，J＝8.0Hz，1H)，10.25(s，1H)，13.10-13.80(brs，1H)。
MS(ESI -)m/z258(M-1)
Embodiment 21.2:3-hydroxyl-10-morpholine-4-base-4-oxo-4, the preparation of 6-dihydro-Mi Dingbing [l, 2-b] indazole-2-carboxylate methyl ester
By adopting the program of in embodiment 21.1, describing and utilizing 2-fluoro-6-morpholine-4-base-benzonitrile, prepared the ester of expectation as starting raw material.
1H NMR (300MHz, D6-DMSO): δ 3.30 (m, 4H (hidden)), 3.88 (t by the water peak, J=4.5Hz, 4H), 3.92 (s, 3H), 6.67 (d, J=7.9Hz, 1H), 6.98 (d, J=8.1Hz, 1H), 7.55 (t, J=8.0Hz, 1H), 10.15 (s, 1H), and 13.20-13.55 (brs, 1H)
MS(ESI -)m/z?343(M-1)
Embodiment 21.3:3-hydroxyl-10-morpholine-4-base-4-oxo-4, the preparation of 6-dihydro-Mi Dingbing [l, 2-b] indazole-2-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D01212
Be incorporated in product in the methyl alcohol (15mL) from embodiment 21.2 (172mg, 0.5mmol), sodium methylate (54mg, 1.0mmol) and the 4-flunamine (1.87mg, 1.5mmol), then reflux and stir under heated overnight.Mixture is cooled to room temperature and collects resulting solid by filtering, be dissolved in then in the methylene dichloride (30mL).With aqueous hydrochloric acid (2.0M), water washing solution, dry and concentrated under vacuum, so that desired compounds (84mg, 38.4%) to be provided.
1H?NMR(300MHz，D6-DMSO)：δ13.61(1H，s，NH)，11.97(1H，s，OH)，8.45(1H，t，J＝6.0Hz，NHCH 2)，7.56(1H，t，J＝6.0Hz，Ar-CH)，7.45(2H，dd，J＝9.0，8.0Hz，Ar-CH)，7.20(2H，dd，J＝9.0，9.0Hz，Ar-CH)，7.00(1H，d，J＝8.1Hz，Ar-CH)，6.71(1H，d，J＝7.8Hz，Ar-CH)，4.61(2H，d，J＝6.0Hz，NHCH 2)，3.69(4H，m，CH 2OCH 2)，3.15(4H，m，CH 2NCH 2)。
MS(ESI -)m/z436(M-1)
HPLC Method 798.9%/13.6 minute
Embodiment 21.4:3-hydroxyl-4-oxo-4, the preparation of 6-dihydro-Mi Dingbing [1,2-b] indazole-2-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D01221
By adopting the program of in embodiment 21.3, describing, prepared desired compounds.
1H?NMR(300MHz，DMSO)δ4.55(2H，d，J＝6.6Hz，-NH-CH 2-)，7.18(2H，m，ArH)，7.42(4H，m，ArH)，7.70(1H，t，J＝7.2，7.8Hz，ArH)，8.09(1H，d，J＝7.8Hz，ArH)，9.67(1H，t，J＝6.6Hz，O＝C-NH-CH 2)，12.40(1H，s，OH)。
MS(ESI -)m/z351(M-1)
HPLC Method 796.4%/13.9 minute
Embodiment 21.5:3-hydroxyl-10-morpholine-4-base-4-oxo-4,6-dihydro-Mi Dingbing [l, 2-b] indazole-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
By adopting the program of in embodiment 21.3, describing, prepared desired compounds.
1H?NMR(300MHz，DMSO)δ3.19(4H，s，N-CH 2-CH 2-O)，3.75(4H，s，N-CH 2-CH 2-O)，4.63(2H，d，J＝6.3Hz，-NH-CH 2-)，6.70(1H，dd，J＝7.8Hz，ArH)，7.00(1H，dd，J＝8.4Hz，ArH)，7.41(1H，dd，J＝6.9，2.1Hz，ArH)，7.56(1H，t，J＝8.2Hz，ArH)，7.66(2H，m，ArH)，8.59(1H，t，J＝5.7Hz，O＝C-NH-CH 2)，11.81(1H，s，OH)。
MS(ESI -)m/z486(M[Cl 35]-1)
HPLC Method 794.3%/15.8 minute
Embodiment 21.6:3-hydroxyl-6-methyl-4-oxo-4, the preparation of 6-dihydro-Mi Dingbing [l, 2-b] indazole-2-carboxylate methyl ester
Figure G2007800515627D01231
Step 1:
Mix the amino benzopyrazoles of 3-(266mg, 2mmol) and Tetra hydro Phthalic anhydride (296mg 2mmol) and at 170 ℃ heated 30 minutes down.Mixture is cooled to room temperature, adds methyl alcohol (10mL) thereafter, (surpassed) the sonication mixture then 2 minutes.By solid collected by filtration and with the product (352mg, 67%) of methanol wash so that expectation to be provided.
1H?NMR(300MHz，D6-DMSO)δ7.13-7.20(m，1H)，7.40-7.48(m，1H)，7.60-7.66(m，1H)，7.70(dd，J＝8.2，0.8Hz，1H)，7.94-8.07(m，4H)，13.44(s，1H)。
Step 2:
At room temperature with methyl iodide (1.41g, 10mmol) be added drop-wise to product in DMF (50mL) from step 1 (2.63g, 10mmol) and salt of wormwood (2.76g is in stirred solution 20mmol).After 3 hours, reaction mixture is cooled to room temperature and injects frozen water (300mL), (3 * 100mL) extract to use methylene dichloride then.With the organic layer that the salt water washing merges, dry and vapourisation under reduced pressure is then by the resulting residue of re-crystallizing in ethyl acetate, so that desired compounds (2.27g, 82%) to be provided.
1H?NMR(300MHz，D6-DMSO)δ4.12(s，3H)，7.19(ddd，J＝8.0，7.0，0.8Hz，1H)，7.49(ddd，J＝8.6，6.9，1.1Hz，1H)，7.71(dt，J＝8.0，1.1Hz，1H)，7.73(dt，J＝8.8，0.9Hz，1H)，7.94-8.06(m，4H)。
Step 3:
Will (277mg, (588mg be in mixture 10mmol) 1mmol) to be suspended in methyl alcohol (15mL) and 85% moisture hydrazine hydrate from the product of step 2.Heated mixt is 1 hour under refluxing, and is cooled to room temperature then.Add entry (40mL) and use methylene dichloride (3 * 20mL) extraction mixtures.With the organic layer that the salt water washing merges, dry and concentrated under vacuum.By flash chromatography (methylene chloride 10: 1) purifying residue so that desired compounds (105mg, 72%) to be provided.
1H?NMR(300MHz，DMSO)δ3.71(s，3H)，5.39(s，2H)6.85-6.93(m，1H)，7.21-7.33(m，2H)，7.66(dt，J＝8.0，1.0Hz，1H)。
Step 4:
The program of describing in embodiment 3 is used for product from step 3 so that desired compounds to be provided.
1H?NMR(300MHz，D6-DMSO)δ3.87(s，3H)，3.90(s，3H)，7.35-7.47(m，1H)，7.76(d，J＝3.5Hz，2H)，8.04(d，J＝8.0Hz，1H)，10.35(s，1H)。
MS(ESI +)m/z296(M+23)
Embodiment 21.7:3-hydroxyl-6-methyl isophthalic acid 0-morpholine-4-base-4-oxo-4, the preparation of 6-dihydro-Mi Dingbing [l, 2-b] indazole-2-carboxylate methyl ester
Figure G2007800515627D01241
By utilizing 4-morpholine-4-base-1H-indazole-3-base amine and adopting the program of in embodiment 21.6, describing, prepared desired compounds.
1H NMR (300MHz, D6-DMSO) δ 3.30 (4H, hidden) by the water peak, 3.82 (s, 3H), 3.84-3.93 (m, 7H), 6.79 (d, J=8.0Hz, 1H), 7.23 (d, J=8.1Hz, 1H), 7.61 (t, J=8.2Hz, 1H), 10.24 (s, 1H)
MS(ESI +)m/z381(M+23)
Embodiment 21.8:3-hydroxyl-6-methyl isophthalic acid 0-morpholine-4-base-4-oxo-4, the preparation of 6-dihydro-Mi Dingbing [l, 2-b] indazole-2-carboxylic acid 4-fluoro-benzyl acid amides
By adopting the program of in embodiment 21.3, describing, prepared desired compounds.
1H?NMR(300MHz，D6-DMSO)δ3.15(4H，s，N-CH 2-CH 2-O)，3.66(4H，s，N-CH 2-CH 2-O)，4.62(2H，d，J＝6.0Hz，-NH-CH 2-)，6.70(1H，dd，J＝7.8Hz，ArH)，7.21(3H，m，ArH)，7.45(2H，m，ArH)，7.62(1H，t，J＝8.1Hz，ArH)，8.39(1H，t，J＝6.3Hz，O＝C-NH-CH 2)，12.02(1H，s，OH)。
MS(ESI -)m/z450(M-1)
HPLC Method 799.7%/12.6 minute
Embodiment 21.9:3-hydroxyl-6-methyl isophthalic acid 0-morpholine-4-base-4-oxo-4,6-dihydro-Mi Dingbing [l, 2-b] indazole-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D01252
By adopting the program of in embodiment 21.3, describing, prepared desired compounds.
1H?NMR(300MHz，D6-DMSO)δ3.23(4H，bs，-NCH 2CH 2O-)，3.73(4H，bs，-NCH 2CH 2O-)，3.82(3H，s，-NCH 3)，4.63(2H，d，J＝6.6Hz，-(O＝C)NHCH 2-)，6.84(2H，d，J＝8.1Hz，ArH)，7.27(2H，d，J＝8.1Hz，ArH)，7.39(2H，dd，J＝2.4，8.0Hz，ArH)，7.65(3H，m，ArH)，9.73(1H，t，J＝6.6Hz，-(O＝C)NHCH 2-)，11.87(1H，s，OH)。
MS(ESI -)m/z524(M[Cl 35]+Na)
HPLC Method 796.0%/14.2 minute
Embodiment 21.10:3-hydroxyl-6-methyl-4-oxo-4, the preparation of 6-dihydro-Mi Dingbing [l, 2-b] indazole-2-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D01261
By adopting the program of in embodiment 21.3, describing, prepared desired compounds.
1H?NMR(300MHz，D6-DMSO)δ3.84(3H，s，-NCH 3)，4.55(2H，d，J＝6.0Hz，-(O＝C)NHCH 2-)，7.18(2H，m，ArH)，7.47(3H，m，ArH)，7.76(2H，dd，J＝1.5，9.1Hz，ArH)，8.06(1H，dd，J＝0.9，8.5Hz，ArH)，9.68(1H，bt，-(O＝C)NHCH 2-)，12.47(1H，bs，OH)。
MS(ESI -)m/z365(M-1)
HPLC Method 785.0%/12.8 minute
Embodiment 21.11:3-hydroxyl-6-methyl-4-oxo-4,6-dihydro-Mi Dingbing [l, 2-b] indazole-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D01262
By adopting the program of in embodiment 21.3, describing, prepared desired compounds.
1H?NMR(300MHz，D6-DMSO)δ3.84(3H，s，-NCH 3)，4.56(2H，d，J＝6.6Hz，-(O＝C)NHCH 2-)，7.37(2H，dd，J＝2.1，8.4Hz，ArH)，7.47(1H，m，ArH)，7.62(2H，m，ArH)，7.77(2H，d，J＝3.6Hz，ArH)，8.06(1H，d，J＝8.1Hz，ArH)，9.73(1H，bs，-(O＝C)NHCH 2-)，12.34(1H，bs，OH)。
MS(ESI -)m/z415(M[Cl 35]-1)，417(M[Cl 37]-1)
HPLC Method 788.0%/14.4 minute
Embodiment 22: the preparation of dibasic 3-hydroxyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid benzyl acid amides
Embodiment 22.1:3-hydroxyl-9-iodo-7-morpholine-4-base-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D01271
Step 1:
(1.53g 7.16mmol) joins that (2.0M, 30mL) (2.0g is heated to 100 ℃ in stirred solution 17.9mmol) and with reaction to the 5-fluoro-2-aminopyridine in aqueous sulfuric acid with sodium periodate.Will (2.68g, drips of solution 17.9mmol) be added in the reaction mixture at the sodium iodide in the water (10mL).After finishing adding, backflow mixture 1 hour is cooled to room temperature then.Drip saturated sodium bicarbonate aqueous solution to regulate pH, use methylene dichloride (3x) to extract mixture then to about 7.0.With the organic layer that aqueous solution of sodium bisulfite, salt water washing merge, dry and concentrated under vacuum.Residue is carried out column chromatography (hexane/ethyl acetate 4: 1) so that desired compounds (2.56g, 60%) to be provided.
1H?NMR(300MHz，CDCl 3)δ4.70-5.03(brs，2H)，7.69(dd，J＝7.2，2.1Hz，1H)，7.93(d，J＝2.1Hz，1H)
MS(ESI +)m/z?239(M+1)
Step 2:
Will (400mg 1.68mmol) be dissolved in the vitriol oil (2mL) and be cooled to-10 ℃ from the product of step 1.In this stirred solution, drip 30% aqueous hydrogen peroxide (2.3g, 20.2mmol) and the mixture of the vitriol oil (4.2mL).With this mixture remain on-10 ℃ following 30 minutes, be heated to 8 ℃ and under this temperature, stir and spend the night thereafter.Mixture is injected frozen water (50mL) to be gone up and extracts with methylene dichloride (3x).With the organic layer that aqueous solution of sodium bisulfite, salt water washing merge, dry and concentrated under vacuum.Residue is carried out the product (36mg, 8.0%) of column chromatography (hexane/ethyl acetate 5: 1) so that expectation to be provided.
1H?NMR(300MHz，CDCl 3)δ8.13(dd，J＝6.9，2.3Hz，1H)，8.35(d，J＝2.4Hz，1H)。
Step 3:
By adopting the program of in embodiment 2.3, describing, obtained desired compounds.
1H?NMR(300MHz，CDCl 3)δ3.36(t，J＝4.9Hz，4H)，3.88(t，J＝4.8Hz，4H)，7.69(d，J＝2.6Hz，1H)，8.06(d，J＝2.6Hz，1H)。
MS(ESI +)m/z358(M+23)
Step 4:
At N 2Will (607mg 1.8mmol) be dissolved in the dehydrated alcohol (50mL) from the product of step 3 under the atmosphere.(2.75g 14.5mmol) drips with 2-3 to add anhydrous stannic chloride (IV) successively.The backflow mixture overnight concentrates under vacuum thereafter.Residue is mixed with water, add aqueous sodium hydroxide solution (0.2M) then to regulate pH to about 11.Extract the mixture of gained with methylene dichloride (3x), and the organic layer that merges with the salt water washing, dry also concentrated under vacuum.Residue is carried out the product (489mg, 89%) of column chromatography (hexane/ethyl acetate 1: 2) so that expectation to be provided.
1H?NMR(300MHz，CDCl 3)δ2.94-3.03(m，4H)，3.77-3.87(m，4H)，4.50-4.76(brs，2H)，7.56(d，J＝2.6Hz，1H)，7.78(d，J＝2.6Hz，1H)
MS(ESI +)m/z?306(M+1)。
Step 5:
The program of describing in embodiment 2 is used for product from step 4 so that desired compounds to be provided.
1H?NMR(300MHz，DMSO-d 6)δ3.14-3.22(m，4H)，3.70-3.81(m，4H)，3.89(s，3H)，8.00(d，J＝2.6Hz，1H)，8.47(d，J＝2.6Hz，1H)，10.31(s，1H)
Step 6:
The program of describing in embodiment 6 is used for product from step 5 so that desired compounds to be provided.
1H?NMR(300MHz，DMSO-d 6)δ3.14-3.21(m，4H)，3.72-3.81(m，4H)，4.62(d，J＝6.6Hz，2H)，7.39(dd，J＝8.3，1.8Hz，1H)，7.61-7.67(m，2H)，8.01(d，J＝2.6Hz，1H)，8.50(d，J＝2.6Hz，1H)，8.95(t，J＝6.5Hz，1H)，11.82(s，1H)。
MS(ESI -)m/z573(M[Cl 35]-1)
HPLC Method 792.7%/12.4 minute
Embodiment 22.2:[2-(4-fluoro-benzylamino formyl radical)-3-hydroxyl-7-iodo-4-oxo-4H-pyrido [l, 2-α] pyrimidine-9-yl]-preparation of urethanum
Figure G2007800515627D01291
Step 1:
In ice bath the cooling in pyridine (15mL) 2,3-diamino-5-iodine pyridine (2.35g, 10mmol).In above-mentioned stirred solution, drip Vinyl chloroformate (1.08g, 10mmol).Under 0 ℃, stirred the mixture 15 minutes, at room temperature stirred then 3 hours, water (30mL) and ethyl acetate (30mL) dilution thereafter.Wash organic phase with water, dry and concentrated under vacuum.Residue is carried out column chromatography (methylene dichloride) so that desired compounds (2.52g, 82%) to be provided.
1H?NMR(300MHz，DMSO-d 6)δ1.24(t，J＝7.1Hz，3H)，4.12(q，J＝7.1Hz，2H)，6.05(s，2H)，7.87(d，J＝2.1Hz，1H)，7.92(s，1H)，8.78(s，1H)。
MS(ESI +)m/z308(M+1)
Step 2:
The program of describing in embodiment 2 is used to provide the product of expectation.
1H?NMR(300MHz，DMSO-d 6)δ1.28(t，J＝7.1Hz，3H)，3.90(s，3H)，4.23(q，J＝7.0Hz，2H)，8.19(d，J＝1.7Hz，1H)，8.56(d，J＝1.8Hz，1H)，8.70(s，1H)10.66(s，1H)
MS(ESI +)m/z?456(M+23)
Step 3:
The program of describing in embodiment 6 is used to provide the product of expectation.
1H?NMR(300MHz，DMSO-d 6)δ1.30(t，J＝7.1Hz，3H)，4.26(q，J＝7.1Hz，2H)，4.62(d，J＝6.1Hz，2H)，7.20(t，J＝9.0Hz，2H)，7.38(dd，J＝8.8，5.4Hz，2H)，8.39(d，J＝I.7Hz，1H)，8.56(d，J＝1.8Hz，1H)，9.99(s，1H)，10.47(t，J＝6.3Hz，1H)，12.66(s，1H)
MS(ESI -)m/z?525(M-1)
HPLC Method 790.4%
Embodiment 23: the preparation of the 7-benzyl of replacement-3-hydroxyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid and acid amides
The preparation of embodiment 23.1:3-(tertiary butyl-dimethyl-silicon alkoxyl group (silanyloxy))-7-iodo-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylate methyl ester
Figure G2007800515627D01311
Step 1:
Begin and adopt in embodiment 5 program of describing by 2-amino-5-iodine pyridine, obtained the ester of expectation.
1H?NMR(300MHz，DMSO-d 6)δ3.85(s，3H)，7.37(d，J＝9.1Hz，1H，H9)，7.79(dd，J＝9.3，2.1Hz，1H，H8)，8.86(d，J＝2.1Hz，1H，H6)，8.50(d，J＝2.6Hz，1H)，10.46(s，1H，OH)。
MS(ESI +)m/z347(M+1)
Step 2:
(program of describing in the step 1) has obtained the silyl compound of expecting at embodiment 13.7 by adopting.
1H?NMR(300MHz，D6-DMSO)：δ0.25(s，6H)，0.93(s，9H)，3.85(s，3H)，7.44(dd，J＝9.2，0.8Hz，1H)，7.94(dd，J＝9.3，1.9Hz，1H)，8.97(dd，J＝1.9，0.8Hz，1H)
The preparation of embodiment 23.2:3-(tertiary butyl-dimethyl-silicon alkoxyl group)-7-(3-chloro-2-fluoro-benzyl)-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylate methyl ester
Figure G2007800515627D01312
Except replacing with tri-o-tolyl phosphorous acid three furans-2-base-phosphorous acid, prepare desired compounds by adopting the program of in WO2004046115, describing.
1H?NMR(300MHz，CDCl 3)：δ0.33(s，6H)，1.00(s，9H)，3.97(s，3H)，4.03(s，2H)，7.01-7.14(m，2H)，7.29-7.43(m，2H)，7.61(d，J＝9.1Hz，1H)，8.75(d，J＝1.3Hz，1H)。
The preparation of embodiment 23.3:7-(3-chloro-2-fluoro-benzyl)-3-hydroxyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid
(0.5M, (22mg is in stirred solution 0.046mmol) 1.1mL) to join the product from embodiment 23.2 in methyl alcohol (5mL) with aqueous NaOH.Under 50 ℃, stirred the mixture 24 hours.Drip then aqueous hydrochloric acid (1.0M) with pH regulator to 3-4.Vapourisation under reduced pressure methyl alcohol also passes through to filter and collects resulting solid, and is dry under vacuum then, so that the desired compounds as brown solid (13mg, 81%) to be provided.
1H?NMR(300MHz，D6-DMSO)：δ8.73(1H，s，Ar-CH)，7.78(1H，d，J＝9.3Hz，Ar-CH)，7.72(1H，d，J＝9.3Hz，Ar-CH)，7.49(1H，dd，J＝7.5，6.6Hz，Ar-CH)，7.35(1H，dd，J＝7.5，5.7Hz，Ar-CH)，7.20(1H，dd，J＝7.8，7.2Hz，Ar-CH)，4.18(2H，s，CH 2Ar)。
MS(ESI -)m/z347(M[Cl 35]-1)
HPLC Method 796.1%/13.2 minute
The preparation of embodiment 23.4:7-(3-chloro-2-fluoro-benzyl)-3-hydroxyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D01322
Be used to obtain desired compounds from the product 4-flunamine of embodiment 23.3 and by adopting the program of in embodiment 6, describing.
1H?NMR(300MHz，DMSO)δ4.13(2H，s，Cl，F-Ph-CH 2-)，4.48(2H，d，J＝6.3Hz，-(O＝C)NHCH 2-)，7.17(4H，m，ArH)，7.39(3H，m，ArH)，7.50(3H，m，ArH)，9.64(1H，s，ArH)，9.68(1H，t，J＝6.0Hz，-(O＝C)NHCH 2-)，12.21(1H，s，OH)
MS(ESI -)m/z454(M[Cl 35]-1)
HPLC Method 794.0%/18.1 minute
Embodiment 23.5:7-(3-chloro-2-fluoro-benzyl)-3-hydroxyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D01331
Be used to the product 3 from embodiment 23.3,4-dichloro-benzylamine and the program of describing in embodiment 6 by employing have obtained desired compounds.
1H?NMR(300MHz，D6-DMSO)δ4.13(2H，s，Cl，F-Ph-CH 2-)，4.49(2H，d，J＝6.0Hz，-(O＝C)NHCH 2-)，7.17(4H，t，J＝6.9Hz，ArH)，7.32(2H，m，ArH)，7.59(7H，m，ArH)，8.65(1H，s，ArH)，9.74(1H，t，J＝6.6Hz，-(O＝C)NHCH 2-)，12.08(1H，s，OH)。
MS(ESI +)m/z506(M[Cl 35]+1)
HPLC Method 799.0%/18.0 minute
The preparation of embodiment 24.1:2-[3-(4-fluoro-benzyl)-[l, 2,4] oxadiazole-5-yl]-3-hydroxyl-7-morpholine-4-base-pyrido [1,2-α] pyrimidin-4-one
Step 1:
Used at J.Med.Chem.1999,42 (20), the program of describing among the 4088-4098.
Step 2:
Be used to from the product of embodiment 2.3 and adopt embodiment 8.1 and the program of embodiment 8.2, prepared 3-benzyloxy-7-morpholine-4-base-4-oxo-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid.Under nitrogen atmosphere, (300mg 0.79mmol), triphenyl phosphorous acid (619mg) and triethylamine (0.3mL) merge, at room temperature stirred 10 minutes then with this compound (159mg) and the product from step 1 in acetonitrile (30mL).Dropping tetracol phenixin (0.4mL) also stirred the mixture 11 hours.Under vacuum,, use the salt water washing, dry and evaporation except that desolvating and residue being dissolved in the methylene dichloride.Residue is carried out column chromatography (hexane/ethyl acetate 1: 4) and obtains desired compounds (280mg, 67%) as yellow solid.
1H?NMR(300MHz，D6-DMSO)：δ3.18-3.29(m，4H)，3.40(s，2H)，3.74-3.86(m，4H)，5.17(s，2H)，6.29-6.72(m，2H)，7.11(t，J＝8.8Hz，2H)，7.24-7.46(m，7H)，7.71(d，J＝9.7Hz，1H)，8.05(dd，J＝9.8，2.4Hz，1H)，8.22(d，J＝2.3Hz，1H)
Step 3:
Product from step 2 (260mg, 0.49mmol) the solution 12 hour of heating in toluene (30mL) under refluxing.Reaction mixture is cooled to room temperature and under reduced pressure concentrated.The purifying that is undertaken by column chromatography (hexane/ethyl acetate 1: 4) provides the desired compounds as yellow solid (183mg, 73%).
1H?NMR(300MHz，D6-DMSO)：δ3.27(t，J＝4.7Hz，4H)，3.80(t，J＝4.7Hz，4H)，4.22(s，2H)，5.21(s，2H)，7.17(t，J＝8.9Hz，2H)，7.25-7.44(m，7H)，7.76(d，J＝9.6Hz，1H)，8.07(dd，J＝9.8，2.6Hz，1H)，8.20(d，J＝2.4Hz，1H)
Step 4:
The program of describing in embodiment 19.8 is used to provide desired compounds.
1H?NMR(300MHz，D6-DMSO)：δ10.67(1H，s，OH)，7.97(1H，s，Ar-CH)，7.84(1H，d，J＝7.8Hz，Ar-CH)，7.62(1H，d，J＝9.0Hz，Ar-CH)，7.42(1H，d，J＝8.4Hz，Ar-CH)，7.39(1H，d，J＝8.7Hz，Ar-CH)，7.19(1H，d，J＝8.4Hz，Ar-CH)，7.16(1H，d，J＝9.0Hz，Ar-CH)，4.23(2H，s，ArCH 2)，3.78(4H，m，CH 2OCH 2)，3.20(4H，m，CH 2NCH 2)。
MS(ESI +)m/z424(M+1)
HPLC Method 791.7%/12.0 minute
Embodiment 24.2:2-[3-(3,4-two chloro-benzyls)-[l, 2,4] oxadiazole-5-yl]-preparation of 3-hydroxyl-7-morpholine-4-base-pyrido [l, 2-α] pyrimidin-4-one
Figure G2007800515627D01351
By 3,4-dichloro benzyl nitrile begins and adopt the program of describing in embodiment 24.1, has obtained desired compounds.
1H?NMR(300MHz，D6-DMSO)：δ10.71(1H，s，OH)，7.98(1H，s，Ar-CH)，7.85(1H，d，J＝9.6Hz，Ar-CH)，7.66(3H，m，Ar-CH)，7.36(1H，d，J＝8.1Hz，Ar-CH)，4.24(2H，s，ArCH 2)，3.78(4H，m，CH 2OCH 2)，3.22(4H，m，CH 2NCH 2)。
MS(ESI +)m/z474(M[Cl 35]+1)
Embodiment 25.1:2-[4-(4-fluoro-benzyl)-4,5-dihydro-oxazoles-2-yl]-preparation of 3-hydroxyl-7-morpholine-4-base-pyrido [l, 2-α] pyrimidin-4-one (sodium salt)
Step 1:
Will the product in the ethanol (15mL) from embodiment 2.3 (305mg, 1mmol) and 2-amino-3-(4-fluoro-phenyl)-third-1-alcohol (169mg, stirring the mixture 1mmol) is heated to refluxed 2 days.The vaporising under vacuum solvent is to provide crude product, and it is directly used in next step.
Step 2:
With methylsulfonyl chloride (228mg, 2.0mmol) and triethylamine (0.5mL, 3.59mmol) join in methylene dichloride (50mL) from the stirring the mixture of the product of step 2.After 2 hours, to use ethyl acetate (50mL) diluted reaction mixture, and wash organic phase with saturated aqueous sodium bicarbonate (50mL), water (50mL) and salt solution (50mL), drying is filtered also concentrated under vacuum.By flash chromatography (hexane/ethyl acetate 1: 5) purifying residue so that desired compounds (250mg, 50%, two step) to be provided.
1H?NMR(300MHz，DMSO-d 6)δ2.82(dd，J＝13.8，6.7Hz，1H)，2.97(dd，J＝14.0Hz，6.8Hz，1H)，3.25-3.31(m，4H)，3.50(s，3H)，3.76-3.85(m，4H)，4.01-4.13(m，1H)，4.40-4.48(m，1H)，4.50-4.63(m，1H)，7.12(t，J＝9.0Hz，2H)，7.37(dd，J＝8.6，5.7Hz，2H)，7.82(d，J＝9.5Hz，1H)，8.18(dd，J＝9.7，2.6Hz，1H)，8.23(d，J＝2.4Hz，1H)
MS(ESI +)m/z?503(M+1)
Step 3:
In methyl alcohol (25mL), mix from the product (228mg) of step 3 and solid sodium hydroxide (40mg, 1mmol).At room temperature stir resulting mixture 1 hour, and added frozen water (100mL) then.By filter to collect resulting throw out and with cold water washing so that desired compounds to be provided, as sodium salt (170mg, 76%).
1H NMR (300MHz, D6-DMSO) δ 3.07 (2H, m ,-CH 2-Ph-F), 3.24 (4H, m ,-OCH 2CH 2N-), 3.85 (4H, m ,-OCH 2CH 2N-), 4.54 (1H, m, ring-type-NCHCH 2O-), 7.00 (2H, t, J=9.0Hz, ArH), 7.30 (2H, m, ArH), 7.64 (1H, d, J=10.0Hz, ArH), 7.80 (1H, dd, J=2.7,9.9Hz, ArH), 8.13 (1H, d, J=2.4Hz, ArH).
MS(ESI -)m/z423(M-Na-1)
HPLC Method 787.0%/17.7 minute
Embodiment 25.2:2-[4-(4-fluoro-benzyl)-4,5-dihydro-oxazoles-2-yl]-preparation of 3-hydroxyl-7-morpholine-4-ylmethyl-pyrido [l, 2-α] pyrimidin-4-one (sodium salt)
Figure G2007800515627D01371
Be used to that (product of step 4) also adopts the program of describing in embodiment 25.1, obtained desired compounds from embodiment 13.7.
MS(ESI +)m/z461(M-Na+1)
HPLC Method 785.4%/11.4 minute
1H?NMR(300MHz，D6-DMSO)：δ8.55(1H，s，Ar-CH)，7.42(3H，m，Ar-CH)，6.89(2H，m，Ar-CH)，6.75(1H，m，Ar-CH)，4.27(2H，m，OCH 2CH[N])，3.72(4H，m，CH 2OCH 2)，3.59(2H，s，Ar-CH 2)，3.25(1H，m，OCH 2CH[N])，2.51(4H，m，CH 2NCH 2)
The preparation of embodiment 26.1:7-(l, l-dioxo-isothiazolidine-2-yl)-3-hydroxyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
The preparation of embodiment 26.1.1:3-hydroxyl-7-iodo-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
The program of describing in embodiment 6 is used for that (product of step 1) is to provide desired compounds from embodiment 23.1.
1H?NMR(300MHz，D6-DMSO)：δ4.59(2H，d，J＝6.9Hz，NHCH 2)，7.15(2H，m，ArH)，7.29(1H，d，J＝9.4Hz，H9)，7.38(2H，dd，J＝8.3，5.9Hz，ArH)，7.81(1H，dd，J＝9.4，1.7Hz，H8)，9.71(1H，t，J＝6.9Hz，NHCH 2)，12.33(1H，s，OH)，
MS(ESI +)m/z440(M+1)。
HPLC Method 797.5%/15.5 minute
The preparation of embodiment 26.1.2:3-benzyloxy-7-iodo-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D01382
The program of describing in embodiment 18.1 is used to provide the product of expectation.
1H NMR (300MHz, D6-DMSO) 4.41 (2H, d, J=6.2Hz, NHCH 2), 5.12 (2H, s, ArCH 2O), 7.04 (2H, t, J=9.1Hz, ArH), 7.32-7.39 (7H, m, ArH), 7.50 (1H, dd, J=0.6,9.3Hz, H9), 8.06 (1H, dd, J=2.1,9.3Hz, H8), 9.01-9.13 (2H, m, H6 and NHCH 2)
MS(ESI +)m/z530(M+1)
The preparation of embodiment 26.1.3:3-benzyloxy-7-(l, l-dioxo-isothiazolidine-2-yl)-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
In DMF (4.0mL), mix product (100mg from embodiment 26.1.2,0.189mmol), isothiazolidine 1,1-dioxide (46mg, 0.378mmol), cupric iodide (I) (4mg, 0.019mmol), N, the N-dimethyl-ethylenediamine (3mg, 0.039mmol) and salt of wormwood (55mg, 0.378mmol) and be heated to 80C.After 2 hours, TLC shows that reaction finishes.Reaction mixture is cooled to room temperature and inject aqueous hydrochloric acid (1.0M, 40mL).Collect resulting solid by filtering, and wash with water, drying is carried out column chromatography (methylene chloride 50: 1) then, with the product (93mg, 95%) that expectation is provided.
1H NMR (300MHz, D6-DMSO) δ 3.64 (2H, t, J=7.3Hz, ring-type-(SO 2)-CH 2CH 2CH 2N), 3.89 (2H, t, J=6.5Hz, ring-types-(SO 2)-CH 2CH 2CH 2N-), 4.43 (2H, d, J=5.9Hz, NHCH 2), 5.14 (2H, s, CH 2O), 7.06 (2H, t, J=9.0Hz, ArH), 7.32-7.48 (7H, m, ArH), 7.84 (1H, d, J=9.9Hz, ArH), 8.00 (1H, dd, J=2.8,9.7Hz, ArH), 8.61 (1H, d, J=2.6Hz, ArH), 9.07 (1H, t, J=6.2Hz, NHCH 2).
The preparation of embodiment 26.1.4:7-(l, l-dioxo-isothiazolidine-2-yl)-3-hydroxyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D01392
The program of describing in embodiment 19.8 is used to provide desired compounds.
1H NMR (300MHz, D6-DMSO) δ 3.62 (2H, t, J=7.2Hz, ring-type-(SO 2)-CH 2CH 2CH 2N-), 3.84 (2H, t, J=6.6Hz, ring-types-(SO 2)-CH 2CH 2CH 2N-), 4.50 (2H, d, J=6.3Hz ,-(O=C) NHCH 2-), 7.16 (2H, t, J=SJ Hz, ArH), 7.41 (2H, m, ArH), 7.64 (1H, d, J=9.6Hz, ArH), 7.83 (1H, dd, J=2.7,9.9Hz, ArH), 8.35 (1H, d, J=1.8Hz, ArH), 9.74 (1H, bt ,-(O=C) NHCH 2-), 12.28 (1H, s, OH).
MS(API +)m/z455(M+Na)
Embodiment 26.2:7-(l, l-dioxo-isothiazolidine-2-yl)-3-hydroxyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D01401
The program of describing in embodiment 26.1 is used to provide desired compounds.
1H NMR (300MHz, D6-DMSO) δ 3.62 (2H, t, J=7.8Hz, ring-type-(SO 2)-CH 2CH 2CH 2N-), 3.84 (2H, t, J=63Hz, ring-types-(SO 2)-CH 2CH 2CH 2N-), 4.52 (2H, d, J=6.3Hz ,-(O=C) NHCH 2-), 7.36 (1H, dd, J=2.1,8.1Hz, ArH), 7.62 (3H, m, ArH), 7.84 (1H, dd, J=2.1,9.9Hz, ArH), 8.36 (1H, d, J=2.4Hz, ArH), 9.78 (1H, bt ,-(O=C) NHCH 2-), 12.14 (1H, s, OH).
MS(ESI -)m/z481(M[Cl 35]-1)
The preparation of embodiment 26.3:7-(l, l-dioxo-[l, 2] thiazan-2-yl)-3-hydroxyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D01402
The program of describing in embodiment 26.1 is used to provide desired compounds.
1H?NMR(300MHz，D6-DMSO)：δ12.32(1H，s，OH)，9.72(1H，t，J＝6.3Hz，NH)，8.60(1H，s，Ar-CH)，7.71(1H，d，J＝9.6Hz，Ar-CH)，7.56(1H，d，J＝9.6Hz，Ar-CH)，7.43(2H，m，Ar-CH)，7.18(1H，d，J＝8.7Hz，Ar-CH)，7.15(1H，d，J＝8.7Hz，Ar-CH)，4.51(2H，d，J＝6.3Hz，NHCH 2)，3.77(2H，m，CH 2N)，3.42(2H，m，CH 2S)，2.18(2H，m，CH 2CH 2CH 2S)，1.86(2H，m，CH 2CH 2CH 2S)。
MS(ESI +)m/z447(M+1)
HPLC Method 796.1%/12.0 minute
Embodiment 26.4:7-(l, l-dioxo-[l, 2] thiazan-2-yl)-3-hydroxyl-4-oxo-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D01411
The program of describing in embodiment 26.1 is used to provide desired compounds.
1H NMR (300MHz, D6-DMSO) δ 1.84 (2H, bm, ring-type-(SO 2)-CH 2CH 2CH 2CH 2N-), 2.18 (2H, bm, ring-types-(SO 2)-CH 2CH 2CH 2CH 2N-), 3.40 (2H, bm, ring-types-(S O 2)-CH 2CH 2CH 2CH 2N-), 3.74 (2H, bm, ring-types-(SO 2)-CH 2CH 2CH 2CH 2N-), 4.50 (2H, d, J=6.6Hz ,-(O=C) NHCH 2-), 7.50 (1H, dd, J=1.8,8.2Hz, ArH), 7.58 (3H, m, ArH), 7.69 (1H, dd, J=2.4,9.9Hz, ArH), 8.58 (1H, d, J=I.8Hz, ArH), 9.76 (1H, t, J=6.9Hz ,-(O=C) NHCH 2-), 12.16 (1H, s, OH).
MS(API +)m/z497(M[Cl 35]+1)
HPLC Method 792.0%/13.2 minute
The preparation of embodiment 26.5:3-hydroxyl-4-oxo-7-(2,2,2-three fluoro-kharophens)-4H-pyrido [1,2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
The preparation of embodiment 26.5.1:3-hydroxyl-4-oxo-7-(2,2,2-three fluoro-kharophens)-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
Make under the condition of in embodiment 8.1, describing from the product of embodiment 6 and react, so that the product of expectation to be provided.
1H?NMR(300MHz，D6-DMSO)4.43(2H，d，J＝6.0Hz，NHCH 2)，5.15(2H，s，CH 2O)，7.06(2H，t，J＝8.8Hz，ArH)，7.28-7.51(7H，m，ArH)，7.69(1H，d，J＝9.5Hz，H9)，8.02(1H，dd，J＝1.7，9.6Hz，H8)，9.02(1H，d，J＝1.5Hz，H6)，9.09(1H，t，J＝5.9Hz，NHCH 2)。
MS(ESI +)m/z482(M[Br 79]+1)，484(M[Br 81]+1)
The preparation of embodiment 26.5.2:3-hydroxyl-4-oxo-7-(2,2,2-three fluoro-kharophens)-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 4-fluoro-benzyl acid amides
Figure G2007800515627D01421
Under the condition of in embodiment 26.1.3, describing and utilize trifluoroacetamide, make product reaction, so that desired compounds (22mg, 31%) to be provided from embodiment 26.5.1.
1H?NMR(300MHz，D6-DMSO)δ4.49(2H，d，J＝6.0Hz，-NH-CH 2-)，7.15(2H，m，ArH)，7.42(2H，m，ArH)，7.57(1H，d，J＝9.6Hz，ArH)，7.83(1H，d，J＝9.6Hz，ArH)，9.36(1H，dd，J＝1.8Hz，ArH)，9.72(1H，bt，O＝C-NH-CH 2)，11.96(1H，s，OH)
MS(ESI +)m/z423(M-1)
HPLC Method 796.7%/12.4 minute
Embodiment 26.6:3-hydroxyl-4-oxo-7-(2,2,2-three fluoro-kharophens)-4H-pyrido [l, 2-α] pyrimidine-2-carboxylic acid 3, the preparation of 4-two chloro-benzyl acid amides
Figure G2007800515627D01422
The program of describing in embodiment 26.5 is used to provide desired compounds.
1H?NMR((300MHz，D6-DMSO)δ4.49(2H，bd，J＝6.6Hz，-(C＝O)NHCH 2-)，7.34(1H，m，ArH)，7.60(3H，m，ArH)，7.89(1H，dd，J＝2.4，9.9Hz，ArH)，9.37(1H，d，J＝2.1Hz，ArH)，9.75(1H，bt，-(O＝C)NHCH 2-)，12.14(1H，s，OH).MS(ESI +)m/z?423(M-1)
MS(ESI -)m/z?473(M[Cl 35]-1)
HPLC Method 782.0%/13.7 minute
Embodiment 27.1:[2-(4-fluoro-benzylamino formyl radical)-3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidin-7-yl methyl]-the phosphonic preparation
Step 1:(is used to the product from embodiment 13.7 (step 2))
To the product in toluene (10ml) from embodiment 13.7 (step 2) (347mg, add in stirred solution 0.81mmol) triethyl-phosphite (268mg, 1.62mmol).Heated mixt is 4 hours under refluxing, and is concentrated into drying thereafter under vacuum.Residue is carried out column chromatography (methylene chloride 30: 1) so that desired compounds (373mg, 95%) to be provided.
1H?NMR(300MHz，CDCl3)δ0.33(s，6H)，0.99(s，9H)，1.30(t，J＝7.0Hz，6H)，3.15(d，J＝21.4Hz，2H)，3.98(s，3H)，4.04-4.16(m，4H)，7.55-7.68(m，2H)，8.75(d，J＝3.0Hz，1H)
MS(ESI +)m/z507(M+23)
Step 2:
At room temperature be stirred in product in the methyl alcohol (5mL) from step 1 (115mg, 0.24mmol) and tosic acid (5mg, mixture overnight 0.024mmol).Concentrated solution is quantitatively to provide crude product under vacuum, and it is directly used in next step.
Step 3:
To be dissolved in from the crude product of step 2 in the acetonitrile (4mL), and cooling (ice/water-bath) stirred solution.(191mg 0.97mmol) and after 2 hours, is heated to room temperature with solution, stirs then and spends the night to drip Iodotrimethylsilane.Under vacuum, concentrate then with methyl alcohol quenching reaction mixture.In resulting residue, add acetonitrile (4mL) and also (surpass) sonication mixture 5 minutes.Collect resulting solid by filtering, with acetonitrile washing and dry under vacuum, with the product (62mg, 87%) that expectation is provided.
1H NMR (300MHz, CD 3OD) δ 3.36 (d, overlapping, 2H), 4.08 (s, 3H), 7.94 (d, J=9.4Hz, 1H), 8.13 (d, J=9.2Hz, 1H), 8.95 (s, 1H)
MS(ESI -)m/z313(M-1)
Step 4:
The program of describing in embodiment 6 is used to provide target compound.
1H NMR (300MHz, D6-DMSO) δ 3.07 (2H, d, J=20.7Hz ,-PCH 2Ph-), 4.49 (2H, d, J=5.7Hz ,-(O=C) NHCH 2-), 7.15 (2H, m, ArH), 7.50 (4H, m, H7, H8 and 2 * ArH), 8.58 (1H, s, H6), 9.74 (1H, bs ,-(O=C) NHCH 2-), 12.15 (1H, bs, OH)
MS(ESI -)m/z406(M-1)
Embodiment 27.2:[2-(3,4-two chloro-benzylamino formyl radicals)-3-hydroxyl-4-oxo-4H-pyrido [1,2-α] pyrimidin-7-yl methyl]-the phosphonic preparation
Figure G2007800515627D01441
The program of describing in embodiment 27.1 is used to provide desired compounds.
1H NMR (300MHz, D6-DMSO) δ 2.97 (2H, d, J=21.3Hz, PCH 2), 4.49 (1H, d, J=5.4Hz, CH 2NH), 7.49 (5H, m, H8, H9 and 2 * ArH), 8.58 (1H, bs, H6), 9.75 (1H, bs, CH 2NH), 11.8 (1H, bs, OH)
MS(ESI -)m/z456(M[Cl 35]-1)
Embodiment 28 biological assays
Utilize following determination techniques, can test the biological activity of compound of the present invention: embodiment 28.13 ' processing/chain transfer simultaneous determination:
(the Ovenden et al.Phytochemistry.2004 Dec that can use and deliver; 65 (24): 3255-9.) similarly unite 3 '-processing/chain transfer measures program.This mensuration goes for the dull and stereotyped form (form) in 96 holes.Briefly, 400ng compound to be tested is with 30nM substrate DNA incubation in addition, and it comprises annealing U5LTR sequence oligonucleotides, and this oligonucleotide is marked with digoxigenin (DIG; 5 '-ACTGCTAGAGATTTTCCACACTGACTAAAAGGGTC-DIG-3 ') or vitamin H (5 '-vitamin H-GACCCTTTTAGTCAGTGTGGAAAATCTCTAGCAGT-3 '), make every kind of substrate on opposite strand, have DIG or biomarker.Under 37 ℃, reacted 2 hours, will be incorporated into the plain flat board of strepto-affinity by the product that 3 ' processing and chain transfer activity produce and utilize anti-DIG-alkaline phosphatase conjugation thing and p-nitrophenyl phosphate substrate to be detected.
Embodiment 28.2 chain transfer specific assay:
The chain transfer specific assay has and the similar form of 3 ' processing/chain transfer simultaneous determination (form), difference is that it has used the biotinylation substrate, the LTR end of its expression preprocessing (5 '-vitamin H-GACCCTTTTAGTCAGTGTGGAAAATCTCTAGCA-3 ').
Annealing oligonucleotide 5 in 10mM Tris-Cl pH8.0,100mM NaCl, 2mM EDTA (final concentration is 30 μ M) '-vitamin H-GACCCTTTTAGTCAGTGTGGAAAATCTCTAGCA-3 ' and 5 '-ACTGCTAGAGATTTTCCAC ACTGACTAAAAGGGTC-Dig-3 '
Each reaction (40 μ l) is included in 30nM substrate DNA and the 400ng intergrase in the reaction buffer, and wherein reaction buffer comprises 20mM Tris-Cl pH7.5,25mM NaCl, 5mM MnCl 2, 5mM MgCl 2, 5mM B-ME, 50 μ g/mL BSA, 0.05% (v/v) tween 20.
With 1/10 end reaction volume compound is joined among the DMSO.
37 ℃ of following incubation reaction 2 hours, then add 60 μ l and regulate damping fluid, it comprises 33mM Tris-Cl pH 7.5,664mM NaCl, 16.6mM EDTA, the 0.166mg/mL salmon sperm dna through (surpassing) sonication.
Then with sample transfer to strepto-affinity element coating flat board, and the product that at room temperature makes enzyme reaction was in conjunction with 1 hour.
With dull and stereotyped 3 * 5 minutes of 30mM NaOH, 200mM NaCl, 1mM EDTA washing, use washes (wash) 2:10mM Tris-Cl pH8.0,6mM EDTA, the free BSA washing of 0.1mg/mL nuclease 3 * 5 minutes then.
(Roche 0.1U/mL) joins in each hole and dull and stereotyped 1 hour of 37 ℃ of following incubations will to dilute anti-digoxigenin-Phosphoric acid esterase Fab of 1/2000 then in washes 2 damping fluids.
With dull and stereotyped 3 times of TBS-tween 20 (0.1%) washing, use the TBS washed twice then, then add 100 μ l substrates (the 1mg/mL p-nitrophenyl phosphate in 0.1M Tris, pH 9.8) and incubation flat board up to manifesting enough colors.
The inhibition that embodiment 28.3HIV duplicates
Seed cells in the 96 hole microtiter plates with 50,000 cells/50 μ l/ holes, and in the RF-10 that comprises 2 μ g/mL polybrene salt (RF-10/2).Compound is become 4x final concentration in RF-10/2, and 30 μ L are joined in the cell.Join in each hole virus (40 μ L in comprising the RF-10/2 of 1600pfu) or 40 μ LRF-10/2, be used for negative control and be used to measure the compound cytotoxicity.After 24 hours, 90 other μ L media or the medium that comprises the 1x compound are joined in each hole.After infection the 4th day, remove the medium of 100 μ L from each hole, and replace with the fresh medium of the 100 μ l that are with or without compound.After 48 hours, the level of results supernatant liquor and definite extracellular p24.Dilute supernatant liquor and utilize Vironostika p24 to measure kit measurement p24 level with 1/10,000.Calculate EC 50Produce to the 50% needed concentration that does not have the medicine contrast as suppressing HIV p24.
Figure G2007800515627D01471
Figure G2007800515627D01472
N/A is inapplicable
NT is test not.
++ the value between+expression 0.001 μ M to the 1 μ M
++ represent the value between 1 μ M to the 10 μ M
+ expression is greater than the value of 1 μ M
Embodiment 28.4 is with respect to the active comparison of wild-type and sudden change (Q148K) hiv integrase
Embodiment 28.4.1 chain transfer is measured:
Used and be similar to (the Ovenden et al.Phytochemistry.2004 Dec that delivers; 65 (24): program is measured in chain transfer 3255-9.).Briefly, with 400ng enzyme (wild-type or medicament-resistant mutation type) and compound to be tested and with 30nM substrate DNA incubation in addition.Substrate DNA is used for simulating HIV DNA end, and it has experienced 3 ' terminal processing, and comprises annealed U5LTR sequence oligonucleotides, is marked with digoxigenin (DIG; 5 '-ACTGCTAGAGATTTTCCACACTGACTAAAAGGGTC-DIG-3 ') or vitamin H (5 '-vitamin H-GACCCTTTTAGTCAGTGTGGAAAATCTCTAGCA-3 '), make every kind of substrate on opposite strand, have DIG or biomarker.Under 37 ℃, reacted 1 hour.The product that is produced by chain transfer activity is incorporated into the plain flat board of strepto-affinity and utilizes anti-DIG-alkaline phosphatase enzyme conjugate (or binding substances) and p-nitrophenyl phosphate substrate to be detected.
Fig. 1 shows selected compounds with respect to the chain transfer measurement result of wild-type intergrase with the intergrase that comprises the Q148K sudden change as an example.
Embodiment 28.4.2 mutant enzyme (mutated enzyme):
Utilize the site-directed mutagenesis hiv integrase that suddenlys change in comprising the shuttle vectors (pGEM) of most of HIV-1 gag and pol sequence, to produce the intergrase sequence, it has been published as the integrase inhibitor of resistance being given announcement.These sudden changes include but not limited to the sudden change such as Q148K.Then PCR is carried out in the intergrase coding region and be cloned in the bacterial expression vector.The concrete introducing of expectation sudden change confirms by sequential analysis.Protein is expressed, and purifying also is used for chain transfer mensuration.
In whole specification sheets, individual character " comprises " or is meant such as the distortion of " comprising " and comprises key element, integer or the step of being stated, or the group of a plurality of key elements, a plurality of integer or a plurality of steps, but do not get rid of any other key element, integer or step, or the group of a plurality of key element, a plurality of integer or a plurality of steps.
All publications of mentioning in this manual are incorporated into this paper with way of reference.Its any discussion that has comprised in this manual document, decree, material, device, goods etc. only is for background of the present invention is provided.It should not be considered as admitting that any or all these items form the part on prior art bases or the common general knowledge in the field relevant with the present invention, as it the application's each right of priority the date before Australia or the elsewhere existence.
It should be understood by those skilled in the art that under situation about not departing from, can carry out many variations and/or improvement, as shown in the embodiment the present invention as the spirit or scope of the present invention of general description.Therefore, embodiments of the present invention should be counted as illustrative and not restrictive in all respects.
Reference
Tisler，M.and?Zupet，R.，Organic?Preparations?and?Procedures?International，22(4)，1990，532-534.
Vompe?A.F.&?Turitsyna?N.F.，J.Gen.Chem.of?the?USSR，1957，27，3318-3325.
Martinez-Barrasa?V.，Delgado?F.，Burgos?C.，Garcia-Navio?J.L.，Izquierdo?M.L.&Alvarez-Builla?J.，Tetrahedron，2000，56，2481-2490.
Carceller?R.，Garcia-Navio?J.L.，Izquierdo?M.L.，Alvarez-Builla?J.，Fajardo?M.，Gomez-Sal?P.&Gago?F.，Tetrahedron，1994，50(17)，4995-5012.
Burgos?C.，Delgado?F.，Garcia-Navio?J.L.，Izaquierdo?M.L.&Alvarez-Builla?J.，Tetrahedron，1995，51(31)，8649-8654.
de?la?Rosa?R.，Martinez-Barrasa?V.，Burgos?C.&Alvarez-Builla?J.，Tet.Let.，2000，41，5837-5840.
Behrman?E.J.，Kiser?R.L.，Garas?W.F.，Behrman?E.C.&Pitt?B.M.，J.Chem.Res.(M)，1995，1051-1063.

Claims (14)

1. the compound of Formula I or its medicinal derivative, salt or prodrug,
Wherein:
A condenses in the monocycle that contains azo-cycle or bicyclic aromatic or heteroaromatic moiety;
X is selected from the group of being made up of O and S;
Y is selected from the group of being made up of O and S;
R 1Be 0-3 substituting group, each in the described substituting group is independently selected from by CN, C 1-10Alkyl, C 2-10Alkenyl, C 1-10Alkyl PO 3H 2,-O-C 1-10Alkyl, C 1-10Alkyl NR 3R 4,-O-C 1-10Alkyl NR 3R 4, halogen, NR 3R 4, alkaryl, miscellaneous alkyl aryl, aryl, heteroaryl ,-O-alkaryl, SO 2NR 3R 4The group of forming;
R 3And R 4Be selected from independently of one another by hydrogen, CO 2C 1-4Alkyl, C (O) C 1-4Alkyl, C 1-10Alkyl, C 1-10NR 5R 6,-NH (CO) is NHC (CO) 1-4The group that alkyl is formed; Or R 3And R 4Form 5-7 unit heterocycle with the nitrogen that is connected, described heterocycle comprises 0 to 2 other heteroatoms that is selected from N, O or S, and wherein S can be in S, S (O) or S (O) 2Oxidation state, and wherein said heterocycle at carbon atom or nitrogen-atoms place alternatively by one or more halogen, aryl, C (O) C of being selected from 1-4Alkyl, SO 2C 1-4Alkyl, SO 2H, C 1-4Alkyl, CO 2H, CO 2C 1-4Alkyl, NR 5R 6, C 1-4Alkyl NR 5R 6Substituting group replace;
R 5And R 6Be selected from independently of one another by H and C 1-4The group that alkyl is formed, or R 5And R 6Form 5-7 unit heterocycle with the nitrogen that is connected, described heterocycle comprises 0 to 2 other heteroatoms that is selected from N, O or S, and wherein S can be in S, S (O) or S (O) 2Oxidation state, and wherein said heterocycle at carbon atom or nitrogen-atoms place alternatively by one or more halogen and C of being selected from 1-4The substituting group of alkyl replaces;
Work as R 1Be alkaryl or-during the O-alkaryl, the substituent aromatic yl group of described alkaryl is selected from C alternatively 1-10Alkyl ,-O-C 1-10Alkyl, C 1-10Alkyl NR 3R 4,-O-C 1-10Alkyl NR 3R 4, halogen, NR 3R 4, alkaryl ,-O-alkaryl, SO 2NR 3R 4Substituting group replace,
B do not exist or-C (O)-;
C do not exist or be selected from by-O-,-NH-and-group of NH-NH-C (O)-form;
R 2Be selected from by heteroaryl, heterocyclic radical and R 7The group of forming;
R 7Be selected from H, alkaryl and C 1-10Alkyl;
Condition is if R 2Be R 7, then B and C must exist.
2. compound according to claim 1, wherein, R 2It is heteroaryl.
3. compound according to claim 2, wherein, R 2Replaced by aryl or alkaryl.
4. according to each described compound in the claim 1 to 3, wherein, the compound of described Formula I is selected from the group of being made up of the compound of Formulae II, III, IV, V and VI:
Figure A2007800515620003C1
Figure A2007800515620004C1
Wherein, Z is O, S or NR 8, R wherein 8Be H, C 1-10Alkyl, C 1-10Alkyl NR 3R 4, alkaryl, miscellaneous alkyl aryl, aryl and heteroaryl.
5. according to each described compound in the claim 1 to 3, wherein, the compound of described Formula I is the compound of chemical formula VII:
Figure A2007800515620004C2
6. according to each described compound in the claim 1 to 5, wherein, NR 3R 4It is morpholine.
7. according to each described compound in the claim 1 to 6, wherein, R 7Be luorobenzyl, more preferably 4-luorobenzyl.
8. according to each described compound in the claim 1 to 7, wherein, R 7Be dichloro benzyl, more preferably 3, the 4-dichloro benzyl.
9. according to each described compound in the claim 1 to 8, wherein, heteroaryl is selected from the group of being made up of tetrazolium, triazole, pyrazoles, imidazoles, oxazole, oxadiazole, thiazole, thiadiazoles.
10. compound according to claim 1 has following chemical formula:
Figure A2007800515620005C1
11. a treatment or the method for preventing virus infection among the curee comprise each described compound or its medicinal derivative, salt or prodrug in the claim 1 to 10 that gives described curee's significant quantity.
12. be used for the treatment of or prevent the application in the medicine for treating viral infections among the curee in preparation according to each described compound or its medicinal derivative, salt or prodrug in the claim 1 to 10.
13. method according to claim 10 or application according to claim 12, wherein, described virus infection is that HIV or SIV infect.
14. a pharmaceutical composition comprises according to each described compound and pharmaceutical carrier, thinner or vehicle in the claim 1 to 10.
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