CN101233129A

CN101233129A - Substituted biarylheterocycle derivatives as protein kinase inhibitors for the treatment of cancer and other diseases

Info

Publication number: CN101233129A
Application number: CNA2006800281850A
Authority: CN
Inventors: 扬·扬; 普兰尼·扬; 马丁·米兰达; 麦克拉·普法尔; 布鲁斯·卡特; 穆巴拉克·鲁沙利夫; 马格努斯·普法尔
Original assignee: Pfahl Family Corp (199679)
Current assignee: Pfahl Family Corp (199679)
Priority date: 2005-05-31
Filing date: 2006-05-30
Publication date: 2008-07-30
Also published as: JP2008542382A; EP1896462A2; CA2650999A1; AU2006252629A1; WO2006130613A3; WO2006130613A2

Abstract

The present invention is directed to certain patentable substituted benzoxazole derivatives that exhibit protein kinase (PK) inhibition activity or modulating ability. Pharmaceutical compositions comprising these compounds, and methods for preparing and using them, are also described. For example, these heterocyclic compounds are useful in treating disorders related to abnormal PK activity, including diseases and disorders involving aberrant cell proliferation, for example, AML, CML, gastrointestinal stromal cancers, thyroid cancer, other cancers and leukemias, as well as other diseases such as inflammation and atherosclerosis.

Description

The biarylheterocycle derivatives that replaces is as kinases inhibitor treatment cancer and other diseases

Background of invention

The c-Abl tyrosine kinase inhibitor, the clinical success that ST1571 (Gleevec or Imanitib, Novartis Pharma) is used for the treatment of chronic myelocytic leukemia (CML) has promoted with kinases inhibitor (PKIs) as the development for the targeted therapies of other types leukemia and noumenal tumour.CML is a kind of rare, virulent myeloproliferative disorder, is characterized in having taken place in the hemopoietic stem cell (9: 22) (q34; Ql 1) karyomit(e) (Ph karyomit(e)) transposition, and activation (the Heisterkamp N on the protein tyrosine kinase Bcr-Abl composition, Stam K, Groffen J, de Klein A and Grosveld G 1985 Structuralorganization of the bcr gene and its role in the Ph ' translocation.Nature 315:758-61; Barila D and Superti-FurgaG 1998. this chromosome deficiencies frequency of occurrences in all chronic myelocytic leukemia patients is 95%, and the frequency of occurrences is 30%-50% in adult acute lymphoblastic leukemia patient.Anintramolecular?SH3-domain?interaction?regulatesc-Abl?activity.Nature?Genetics?18：280-2)。CML accounts for 0.4% of all malignant tumours, and epidemiology is that per 10,000 philtrums have 3-5 example (Baker DE 2002 Imatinib Mesylate.NewDrug Review.Reviews in Gastroenterological disorders.2:75-86).

Though suppressing the Abl Tyrosylprotein kinase with Gleevec is effectively for the treatment chronic myelocytic leukemia, is not used in the targeted therapies of the more general acute myeloblastic leukemia of treatment (AML).In addition, though Gleevec is effectively for treatment deterioration and blast crisis, the resistance that occurs is a problem.As a kind of more general disease, AML accounts for all significant proportions that become the human leukemia case, in the U.S. about 7200 victims there is every year, and the number bigger (Seppa N 2002 Smart drugs:Leukemia treatments nearing prime time.Science 161:2-4) in the worldwide.

In the U.S., about 10,000 newly-increased AML cases are arranged every year, then there are 200,000 examples in the whole world.40 percent AML patient has a kind of Flt-3 enzyme of sudden change, this enzyme has the activity of constitutive character like this, the activation and the uncontrollable bone marrow stem cell hyperplasia (Abu-Duhier FM, Goodeve AC, Wilson GA et al.2000) that cause Tyrosylprotein kinase.Flt-3 internal series-connection in adult AML patient repeats sudden change and has formed a kind of high risk population's group (Br.J Haematol 111:190-195).This fatal non-constant of hematologic cancers prognosis, patient's five-year survival rate has only 14%.Therefore, need be to the equal beneficial agents that can treat of early stage and late stage of AML.The Flt-3 kinases is that disease progression is necessary, has therefore also just become a desirable target of development new therapy.

Flt-3 (FMS sample Tyrosylprotein kinase 3) is a kind of receptor tyrosine kinase, the phosphorylation of the oh group of its catalytic proteins tyrosine residues.Receptor tyrosine kinase comprises the transmembrane receptor with different biological function of a big nation.At present, have at least the receptor tyrosine kinase of 19 kinds of different subtribes to be identified.Flt-3 is one of member of platelet-derived somatomedin (PDGFR) receptor tyrosine kinase subtribe, and has identical constitutional features with KIT, FMS and PDGFR.Structurally, Flt-3 has five immunoglobulin like domain at extracellular region, and a single is striden the film sequence, and the nearly membranin of an intracellular weak point, its following and then intracellular kinase structural domain (Levis M, Allebach J, a Tse K-F, ZhengR, Baldwin BR, Douglas Smith B, Jones-Bolin S, Ruggeri B, DionneC and Small D (2002)).

The tyrosine kinase inhibitor of Flt-3 target all has cytotoxicity (Blood 99 (11): 3885-3891) with external in vivo the leukemia cell.Its active signal is transduceed by the protein phosphorylation in the biochemical signals transduction path in autophosphorylation and the born of the same parents, and this has promoted uncontrollable cell proliferation and has suppressed apoptosis.When with after the Flt-3 part combines, wild-type receptor generation dimerization, the phosphorylation of passing through to be produced activates its tyrosine kinase domain.RAS-GAP, PLC, PI3-kinases, STAT5 and ERK1/2 are the signal of interest albumen that is associated with the activation of Flt-3.

The subfamily member of other receptor tyrosine kinase families comprises the EGF receptor tyrosine kinase, insulin receptor tyrosine kinase and FGF receptor tyrosine.For example, the EGF receptor tyrosine kinase all was expression in many cancers in as lung cancer and mammary cancer.The Tarceva (for nonsmall-cell lung cancer) that Herceptin that the Iressa that Astrazeneca company produces, Genentech company produce (to the positive mammary cancer that shifts of HER2-) and OSI-Genentech company produce etc. is the selectively targeted medicine at the EGFR acceptor.Severally in addition just comprise: SU5416 at the small molecules receptor tyrosine kinase inhibitors of clinical development, SU6668 and SUl 1248 (Sugen-Pfizer company), ZD-6474 (Astrazeneca company) and PTK787/ZK222584 (Novartis company), and can suppress multiple kinases such as VEGFR, FGFR, PDGFR and c-kit.In addition, also carried out some and identified small molecules other trials as kinases inhibitor.For example, two monocycles, dicyclo and heterocyclic aromatic compound (PCT WO92/20642), vinyl 7-azaindole derivatives (PCT WO 94/14808) and 1-cyclopropyl-4-arsenic pyridine yl-quinoline ketone (U.S. Patent number No.5,330,992 and compound of styryl I (U.S. Patent number No.5,217,999), arsenic acridine compound (the U.S. Patent number No.5 that styryl replaces, 302,606), quinazoline derivant (Spain patent No. No.0566 266 A1), selenizing indoles and selenide (PCT WO 94/03427), three ring poly aquation oxymetazoline compounds (PCT WO 92/21660), phenmethyl phosphinic acid compounds (PCT WO 91/15495) has all been reported as protein tyrosine kinase inhibitor and has been used for treatment for cancer potentially.In addition, benzene  azoles, benzoxazol and benzimidizole derivatives (PCT WO 02/072543 A2) have also been reported and have been helped cancer and other treatment of diseases, still but are not suitable for as specific kinase whose target inhibitor, and this also is the situation that many other heterogeneous ring compounds illustrate in patent.

The invention relates to the kinase inhibitor that a class can obtain having of patent of specific substituted benzene  oxazole derivatives, Flt-3 is had restraining effect efficiently, can carry the leukemia cell that Flt-3 suddenlys change with external killing and wounding specifically in vivo.In addition, these compounds also can suppress c-Kit (mutant that comprises the Gleevec tolerance) effectively in experiment in vitro, and/or RET, and/or PDGFR β (platelet derived growth factor receptor β), because these kinases are being brought into play important effect to its propagation in certain cancers,, this type of inhibitor is used for these treatment for cancer so can expecting.These compounds structurally are different from the  of the benzene widely oxazole derivatives of PCT WO 02/072543 A2 indication.Those compounds are in the news and have antitumour activity widely because of having significant cytotoxicity.Therefore said compound has good selectivity antitumour activity, and they can kill and wound the leukemia cell system that carries the Flt-3 mutant and not kill and wound other clone in experiment.Therefore, said compound structurally and because have the anticancer activity of selectivity, is different from the compound that WO 02/072543 A2 is reported.Have only target anticancer active rather than widely antitumour activity be an one major advantage because this compound is hopeful to have the side effect and the toxicity of minimum.What specify is that the compound of this vitro inhibition Flt-3 then is to carry the human AML tumor growth of Flt-3 mutant and the inhibitor of deterioration in vivo.In addition, some in these heterogeneous ring compounds also demonstrate the effect with other types kinase inhibitor, and these kinases comprise c-Abl, PDGFR, c-Kit and/or RET, they all are knownly to have participated in matter tumour between CML, stomach and intestine, and/or the generation of thyroid carcinoma.Other that find have AURKA, FGFR3, JAK2, PDGFR β, RET and VEGFR at the external kinases that is suppressed by high-affinity.Like this, the every details of kinase inhibitor that general structure and special example have been contained a kind of novel type among the application, can be as " target anticancer " medicament, and work by the mechanism that limits, suppress specific kinases, and therefore be different from the existing medicine that is used for the treatment of granulocyte leukemia and other cancers significantly.Therefore, said heterogeneous ring compound helps to treat some because of the disease that uncontrollable propagation causes, comprises matter cancer between AML, CML, stomach and intestine, thyroid carcinoma and other cancers and other are as diseases such as inflammation, atherosclerosiss.In addition, because at this compound of analyzing is the optionally kinase inhibitor with good oral bioavailability rate, therefore can expect that these compounds have limited side effect, be applicable to the multiple cancer of treatment, separately or with other medication combined other treatment for cancer that is used for.Therefore, the present invention also relates to the usage that this compound is used for the treatment of solid tumor and other diseases.The pharmaceutical composition that comprises these compounds has also been described, the method for treatment disease and their preparation method.

Summary of the invention

The present invention be directed to the benzoxazol derivative of the replacement of specific obtained patent, these derivatives have protein kinase (PK) and suppress activity or regulating power, therefore help to treat the active unusual obstacle that causes of PK.

An aspect of of the present present invention relates to compound or its isomers, meta-bolites, many types of variant, prodrug or their salt of following chemical formula:

Wherein:

"----" exists or do not exist;

W is (a) or (b)

A is-CR ₂₁R ₂₂-,-NR ₂₃-,-O-or-S-;

B is-OR ₂₄,-SR ₂₅,-NR ₂₈R ₂₉

D and E are-CR jointly or independently ₃₀-or-N-;

Ar is (c) or (d)

And

R ₁, R ₂, R ₃, R ₁₁, R ₁₂, R ₂₁, R ₂₂, R ₂₃, R ₂₄, R ₂₅, R ₂₈, R ₂₉And R ₃₀Be independently or jointly: hydrogen; alkyl; the alkyl that replaces; haloalkyl; thiazolinyl; the thiazolinyl that replaces; alkynyl; the alkynyl that replaces; halogen; cyano group; nitro; hydroxyl; acyl group; the acyl group that replaces; acyloxy; amino; single substituted-amino; disubstituted amido; alkyl sulfonamide; aryl sulfonic acid amides; alkyl urea; the aryl urea; alkyl carbamate; aryl carbamate; heteroaryl; alkoxyl group; the alkoxyl group that replaces; halogenated alkoxy; alkylthio; the sulfo-haloalkyl; carboxyl; carbalkoxy; the alkyl carboxylic acid amides; the alkyl carboxylic acid amides that replaces; the dialkyl group carboxylic acid amides; the perhaps dialkyl group carboxylic acid amides of Qu Daiing.

Another aspect of the present invention provides the synthetic method of these compounds.

On the other hand, the present invention relates to suppress uncontrollable cell proliferation with said compound, for example various types of cancers and leukemia also comprise inflammatory diseases or atherosclerosis therapy.The invention still further relates to the certain methods that suppresses uncontrollable cell proliferation, for example, a kind of cancer or leukemia need this type of inhibitor after diagnosing, and to a Mammals, a preferred people takes medicine.The invention allows for the certain methods of the uncontrollable cell proliferation of treatment, for example cancer and leukemia, comprise and treating making a definite diagnosis a kind of effective dose that a Mammals that suffers from this type of disease takes a kind of compound of the present invention, also comprise the certain methods for the treatment of inflammatory diseases, and use a kind of effective dose of a kind of compound of the present invention to treat making a definite diagnosis a kind of Mammals that suffers from certain inflammatory diseases.

On the other hand, the present invention relates to pharmaceutical composition, comprise the mixture that constitutes with the carrier that allows in one or more pharmacy, auxiliary material etc. and a kind of compound described herein.A related aspect relates to medicine box, this comprises, for example, and an a kind of unit dose packaging (for example vial) in a kind of proper container about pharmaceutical composition of the present invention, this packing may be chosen wantonly and be packaged into box or other similar packings, shipping or storage before being used to use.In a preferred embodiment of the invention, these medicine boxs also comprise a package insert usually, and this specific compound, its usage and the details of purposes are provided.

Brief Description Of Drawings

Fig. 1 illustrates the kinase inhibition characteristic profile of compound 1.

Figure-2 illustrates 2,3,4 and 5 couples of MV4 of compound; Killing and wounding of 11 people AML clones.

Fig. 3 illustrates selecting cell and kills and wounds.Compound 1 kills and wounds AML cell (MV4 specifically; 11) rather than prostate gland (PC-3) and pancreatic cell (Bx-PC-3).

Fig. 4 illustrates the kinase inhibition characteristic profile of

compound

2,5 and 8.

Fig. 5 illustrates compound 2 oral administration biaavailability and pharmacokinetics in the rat body.

Fig. 6 illustrates compound 2 and suppress AML tumour progression situation in the nude mouse model of extracing thymus gland.

Fig. 7 illustrate that compound 2 suppresses to have set up in nude mice model huge AML tumour situation.

The selecting cell that Fig. 8 illustrates compound 5 kills and wounds.

Fig. 9 illustrates compound 5 in intravital oral bioavailability of rat and pharmacokinetics.

Figure 10 illustrates the study on the efficiency of compound 5 in nude mice model.

Figure 11 illustrates compound 2 and compound 5 reduce tumor growth in nude mice model comparison.

Figure 12 illustrates compound 5 restraining effect to the huge tumor set up in nude mice model.

Figure 13 illustrates a kind of representative solution of the route of synthesis of some compounds with Chemical formula 1 (wherein m is 0, and n is 1) disclosed here.

Embodiment

The invention provides a kind of compound that helps to prevent, alleviate or treat in other respects cancer, especially to human and other mammiferous acute myeloblastic leukemias, gastric and intestinal cancer (comprising mutant), specific thyroid carcinoma to the Gleevec tolerance.In addition, proved that these compounds among the present invention have oral administration biaavailability, after taking medicine separately or with a kind of vehicle is oral, in blood plasma, all had very high level because no matter be.Oral administration biaavailability make its can oral administration chronic disease because oral have can the oneself take medicine and reduce the advantage of the expense that other mode administrations produce.

Definition

In specification sheets described here and chemical formula, following term is defined as follows.Except these terms, if necessary, other terms will have been defined at the elsewhere of specification sheets.Unless at this special instruction is arranged in addition, the industry slang that occurs in this specification sheets still has the meaning in the industry that they are generally acknowledged.

" medicament " is meant a kind of a kind of active ingredient that is used for reaching the expection therapeutic purpose and gives.

Term " alkyl " is meant a kind of straight or branched group that comprises 1 to 12 carbon, for example methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, sec-butyl, the tertiary butyl, amyl group, tert-pentyl, n-pentyl etc.Term " thiazolinyl " expression a kind of straight or branched hydrocarbon group that comprises 1 to 12 carbon and carbon-to-carbon double bond, for example vinyl, allyl group, crotyl, 3-butenyl, pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 5-heptenyl, 6-heptenyl etc.Term " thiazolinyl " comprises the dialkylene and the trialkenyl of straight chain and side chain.Term " alkynyl " expression a kind of straight or branched carbohydrate group that comprises 1 to 12 carbon and carbon-to-carbon three key, for example ethynyl, 1-proyl, 2-propynyl, ethyl acetylene base, 2-butyne base, 3-butynyl, 1-pentynyl, valerylene base, 3-pentynyl, 4-pentynyl, 1-hexin base, 2-hexin base, 3-hexin base, 4-hexin base, 5-hexin base etc.Term " alkynyl " comprises diine or three alkynes.

The group that comprises 1 to 12 carbon of a kind of above-mentioned definition of term " alkyl of replacement " expression is by one or more groups; but preferred one; two or three are selected from following group and replace, and comprising: hydroxyl; halogen; cycloalkyl; amino; single substituted-amino; disubstituted amido; acyloxy; nitro; cyano group; carboxyl; the epoxy carbonyl; the alkyl carboxylic acid amides; the alkyl carboxylic acid amides that replaces; the dialkyl group carboxylic acid amides; the dialkyl group carboxylic acid amides that replaces; alkyl sulphonyl; alkyl sulphinyl; alkylthio; the sulfo-alkylhalide group; alkoxyl group; the alkoxyl group that replaces; perhaps halogen alkoxyl group.They can be identical or different when occurring more than a group.

The group that comprises 1 to 12 carbon of a kind of above-mentioned definition of term " thiazolinyl of replacement " expression is by one or more; but preferred one; two or three are selected from following group and replace, and comprising: halogen; hydroxyl; cycloalkyl; amino; single substituted-amino; disubstituted amido; acyloxy; nitro; cyano group; carboxyl; carbalkoxy; the alkyl carboxylic acid amides; the alkyl carboxylic acid amides that replaces; the dialkyl group carboxylic acid amides; the dialkyl group carboxylic acid amides that replaces; alkyl sulphonyl; alkyl sulphinyl; alkylthio; the sulfo-alkylhalide group; alkoxyl group; the alkoxyl group or the halogenated alkoxy that replace.When appearance was substituted more than a group, they can be identical or different.

A kind of group that comprises 1 to 8 above-mentioned definition of term " alkynyl of replacement " expression is replaced but preferred one or two are selected from following group: the alkoxyl group or the halogenated alkoxy of the dialkyl group carboxylic acid amides of the alkyl carboxylic acid amides of halogen, hydroxyl, cycloalkyl, amino, single substituted-amino, disubstituted amido, acyloxy, nitro, cyano group, carboxyl, carbalkoxy, alkyl carboxylic acid amides, replacement, dialkyl group carboxylic acid amides, replacement, alkyl sulfonyl, alkyl sulfenyl, alkylthio, sulfo-alkylhalide group, alkoxyl group, replacement by one or more.

Part (moiety) that contains the cyclic alkyl of 3 to 8 carbon of term " cycloalkyl " expression is above-mentioned definition at this alkyl, comprises groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclopenyl, cyclohexyl, suberyl.The cycloalkyl of a kind of above-mentioned definition of term " cycloalkyl of replacement " expression is further replaced by one or more following radicals, is selected from: the dialkyl group carboxylic acid amides of the alkyl carboxylic acid amides of the alkoxyl group of halogen, alkyl, hydroxyl, alkoxyl group, replacement, carboxyl, carbalkoxy, alkyl carboxylic acid amides, replacement, dialkyl group carboxylic acid amides, replacement, amino, single substituted-amino or disubstituted amido.When cycloalkyl was replaced more than a group, they can be identical or different.

Term " combination therapy " is meant that at least two kinds of different methods of treatment of use reach a kind of treatment plan of set result of treatment.For example, a kind of combined treatment can comprise the medicine of taking two kinds or above different chemical character, as using a kind of compound and the another kind of chemotherapeutics among the present invention.As an alternative, a kind of combined treatment can comprise takes one or more medicines of the present invention, and independent or associating other drug and methods of treatment are as operation, radiation etc.Taking under the situation of two kinds or above different chemical medicine, its activeconstituents can be used as with part a kind of or not compound of the same race and takes as everyone knows.When taking as different compounds, the different activities composition that comprises in the compound can be simultaneously or not simultaneously, through identical or different approach, use identical or different dosage regimens to take, these all depend on different situations and are responsible for doctor's decision.Equally, when implementing together when one or more medicines with such as psychoanalysis etc., this (these) medicine can be before during curee's treatment, among and take afterwards.On the contrary, " monotherapy " is meant a kind of based on taking a kind of treatment plan that the compound of definite curative effect is arranged, and no matter is to take as single dose or long-time multidose.

Term " cycloalkenyl group " expression a group that comprises 3 to 8 carbon, for example cyclopropenyl radical, 1-cyclobutene base, 2-cyclobutene base, 1-cyclopentenyl, 2-cyclopentenyl, 3-cyclopentenyl, 1-cyclohexenyl, 2-cyclohexenyl, 3-cyclohexenyl etc.The cycloalkenyl group of an above-mentioned definition of term " cycloalkenyl group of replacement " expression further is selected from following group and replaces by one or more: the dioxane carboxylic acid amides of the alkyl carboxylic acid amides of the alkoxyl group of halogen, alkyl, hydroxyl, alkoxyl group, replacement, halogenated alkoxy, carboxyl, carbalkoxy, alkyl carboxylic acid amides, replacement, dioxane carboxylic acid amides, replacement, amino, single substituted-amino or disubstituted amido.When cycloalkenyl group was replaced more than a group, they can be identical or different.

An alkyl group in the above-mentioned definition of this used term " alkoxyl group " expression directly combines with an oxygen, as methoxyl group, oxyethyl group, positive propoxy, isopropoxy, n-butoxy, sec-butoxy, isobutoxy etc.The alkoxy grp of an above-mentioned definition of term " alkoxyl group of replacement " expression is by one or more groups; preferred one or two following groups replaces, and is selected from: the alkoxyl group or the halogenated alkoxy of the dialkyl group carboxylic acid amides of the alkyl carboxylic acid amides of hydroxyl, cycloalkyl, amino, single substituted-amino, disubstituted amido, acyloxy, nitro, cyano group, carboxyl, carbalkoxy, alkyl carboxylic acid amides, replacement, dialkyl group carboxylic acid amides, replacement, alkyl sulphonyl, alkyl sulphinyl, alkylthio, sulfo-alkylhalide group, alkoxyl group, replacement.When appearance was substituted more than a group, they can be identical or different.Amino of term " single substituted-amino " expression is selected from following group and replaces: the alkyl of alkyl, replacement or aralkyl, these terms wherein are identical with the explicans in the whole text.

Amino of term " disubstituted amido " expression is replaced by two identical or different following groups, is selected from: the alkyl of the aromatic base of aromatic base, replacement, alkyl, replacement or aralkyl, the term of these terms wherein and definition in the whole text is equivalent in meaning.Some examples comprise dimethylamino, methylethyl amino, diethylin etc.

The alkyl group of an above-mentioned definition of term " alkylhalide group " expression is replaced by the preferred fluorine of one or more halogen, for example trifluoromethyl, pentafluoroethyl group etc.

Term " halogenated alkoxy " represents that the alkoxyl group of an above-mentioned definition directly combines the trifluoromethoxy of formation, five fluorine oxygenation bases etc. with an oxygen.Group that comprises 1 to 8 carbon of term " acyl group " expression is as formyl radical, ethanoyl, propionyl, butyryl radicals, isobutyryl, pentanoyl, caproyl, oenanthyl, benzoyl etc.Direct and oxygen combination of carboxyl groups that comprises 1 to 8 carbon of the above-mentioned definition of term " acyloxy " expression is as acetoxyl group, propionyloxy, butyryl acyloxy, isobutyl acyloxy, benzoyloxy.Aromatic group that comprises 6 to 10 carbon of term " aromatic base " expression comprises phenyl and naphthyl.The aromatic group of an above-mentioned definition of term " aromatic base of replacement " expression is replaced by one or more group, be selected from: hydroxyl, cycloalkyl, aromatic base, the aromatic base that replaces, heteroaryl, heterocycle, the heterocycle that replaces, amino, the single substituted-amino of 1-, disubstituted amido, acyloxy, nitro, cyano group, carboxyl, carbalkoxy, the alkyl carboxylic acid amides, the alkyl carboxylic acid amides that replaces, the dialkyl group carboxylic acid amides, the dialkyl group carboxylic acid amides that replaces, alkyl sulfonyl, alkyl sulfenyl, alkylthio, alkoxyl group, the alkoxyl group or the halogenated alkoxy that replace, wherein these terms here define.Term " halogenation " or " halogen " are meant fluorine, chlorine, bromine or iodine group.

Sulfo-thing group that comprises 1 to 8 carbon, straight or branched of term " alkylthio " expression.Example comprises sulphomethyl, ethyl-sulfide, sulfo-sec.-propyl etc.Alkylthio group of term " sulfo-alkylhalide group " expression is replaced by one or more halogens.Example comprises trifluoromethylthio, 1,1-difluoro thio-ethyl, 2,2,2-trifluorothio ethyl etc.

Term " carbalkoxy " is meant the alkyl ester of a carboxylic acid, and wherein alkyl is identical with definition above.Example comprises methoxycarbonyl, ethoxycarbonyl, the different third oxygen carbonyl etc.Independent alkyl group of term " alkylamino acyl " expression directly combines with the amine of acid amides, and wherein alkyl is identical with definition above.Example comprises N-methyl carboxylic acid amides, N-ethyl carboxylic acid amides, N-sec.-propyl carboxylic acid amides etc.Independent " alkyl of replacement " group of term " the alkyl carboxylic acid amides of replacement " expression as hereinbefore defined, is connected on the amine of acid amides.

Two identical or different alkyl of term " dialkyl group carboxylic acid amides " expression or aralkyl group are connected on the amine of acid amides, and alkyl wherein is identical with definition above.Example comprises N, N-dimethyl carboxylic acid amides, N-methyl-N-ethyl carboxylic acid amides etc.

Two alkyl groups of " dialkyl group carboxylic acid amides " expression that term replaces are connected on the amine of acid amides, and one or two groups here all are " alkyl of replacement " of above definition.As everyone knows, these groups can be identical or different.Example has N, N-dibenzyl carboxylic acid amides, N-benzyl-N-methyl carboxylic acid amides etc.

Carboxyl groups of term " alkylamide " expression is connected on amine or the monoalkylamine, and wherein acyl group is identical with definition above.The example of " alkylamine " comprises monoamide base, propionamido-etc.

Term " aralkyl " is meant a thiazolinyl, as-CH ₂-, one that is above defined has the aromatic yl group that replaces or do not have replacement to replace, and the example of " aralkyl " comprises benzyl and vinylbenzene etc.

A residue of a kind of chemical substance of in the claim of specification sheets and ending place, using, be meant in specific chemical reaction or follow-up synthetic in or the part (moiety) of the synthetic product in the chemical product, no matter whether real from chemical substance, obtaining of this part.Like this, a glycol residue in polyester is represented the one or more-OCH2CH2O-repeating unit in the polyester, and no matter whether ethylene glycol has participated in the synthetic of polyester.Equally, in a kind of chemical compound-2, it is one or more-2 that 4-thiazolidinediones residue refers to, the 4-thiazolidinedione is (moiety) partly, no matter whether this residue is from-2,4-thiazolidinediones reaction generates and obtains in this compound.

According to a kind of " can obtain patent " of the present invention composition, method, machine or make object and be meant that in analyze, this theme satisfies all statutory requirement for patentability.For example, at novelty, unobviousness, etc., if investigation is afterwards found the one or more claims that comprise one or more embodiments and is not supported novelty, unobviousness etc., should (these) claim be subjected to the restriction of " can obtain patent " embodiment definition, get rid of clearly can not obtain patent should (those) embodiment.And, should be interpreted as the wideest zone of reasonableness can be provided in these these appended claims, guarantee its validity again.In addition, if there is the required statutory requirement of one or more patentabilities to be modified, in perhaps being queried to the validity of these appended claims from day of the application's submission or the granted patent, assess the standard whether a concrete statutory requirement satisfy patentability and change, should (these) claim must make explanations with the following methods so: (1) keeps its validity and (2) can provide the wideest zone of reasonableness in the case.

" experimenter " or " patient " is meant that one needs treatment, an animal that can be subjected to the molecules influence among the present invention really.Can comprise vertebrates according to the animal that the present invention treats, such as ox, dog, horse, cat, sheep, pig, and primate (comprising human and inhuman primate) is a more preferred example.

Here must point out that " " of the singulative of using, " one " and " certain " also comprise its plural indicator, unless context has clear and definite explanation in addition in specification sheets and claims.Therefore, as when mentioning " a kind of aromatics ", comprising the mixture of multiple aromatics.

Composition

Some disclosed embodiments of the present invention relate to equation (1):

Wherein

W is

A is-CR ₂₁R ₂₂-,-NR ₂₃-,-O-or-S-;

B is-OR ₂₄,-SR ₂₅,-NR ₂₈R ₂₉

D and E are-CR jointly or independently ₃₀-, or-N-;

L is-CH ₂-;

Ar is a chemical formula (c) or (d)

(c)

M is 0 to 1;

N is 1 to 2;

R ₁, R ₂, R ₃, R ₁₁, R ₁₂, R ₂₁, R ₂₂, R ₂₃, R ₂₄, R ₂₅, R ₂₈, R ₂₉And R ₃₀Be hydrogen independently or jointly; alkyl; the alkyl that replaces; alkylhalide group; thiazolinyl; the thiazolinyl that replaces; alkynyl; the alkynyl that replaces; halogen; cyano group; nitro; hydroxyl; acyl group; the acyl group that replaces; acyloxy; amino; single substituted-amino; disubstituted amido; alkyl sulfonamide; aryl sulfonic acid amides; alkyl urea; the aryl urea; alkyl carbamate; aryl-carbamate; heteroaryl; alkoxyl group; the alkoxyl group that replaces; halogenated alkoxy; thio alkoxy; sulfo-halogen alkoxyl group; carboxyl; carbalkoxy; the alkyl carboxylic acid amides; the alkyl carboxylic acid amides that replaces; the dialkyl group carboxylic acid amides of dialkyl group carboxylic acid amides or replacement; a perhaps one isomers; meta-bolites; many types of variant; prodrug, perhaps salt.The present invention also comprises other forms of compound in the invention, comprises the prodrug form.At this, " prodrug " is meant the compound that comprises one or more functional groups, forms a kind of molecule with therapeutic action in vivo thereby can be removed in vivo or be modified." many types of variant " is meant a kind ofly have compound and compare with another kind of compound, has identical chemical constitution (that is to say, belong to a kind of chemical species) and just has different crystalline structure.

In some embodiments, when W is 2,4-thiazolidinedione and "----" are when existing, and compound is a kind of benzylidene (benzilidine) compound with following structural molecule formula:

(E)

In some embodiments, when W is 2,4-thiazolidinedione and "----" be not when existing, and compound is a kind of benzyl compounds with following structural molecule formula:

(F)

In another embodiment of the invention, when W was (b), compound was a kind of alkyl acetal with following structural molecule formula:

R wherein ₁₁And R ₁₂Be hydrogen independently or jointly, alkyl; the alkyl that replaces; alkylhalide group; thiazolinyl; the thiazolinyl that replaces; alkynyl; the alkynyl that replaces; halogen; cyano group; nitro; hydroxyl; acyl group; the acyl group that replaces; acyloxy; amino; single substituted-amino; disubstituted amido; alkyl sulfonamide; aryl sulfonic acid amides; alkyl urea; the aryl urea; alkyl carbamate; aryl-carbamate; heteroaryl; alkoxyl group; the alkoxyl group that replaces; halogenated alkoxy; thio alkoxy; sulfo-halogen alkoxyl group; carboxyl; carbalkoxy; the alkyl carboxylic acid amides; the alkyl carboxylic acid amides that replaces; the dialkyl group carboxylic acid amides of dialkyl group carboxylic acid amides or replacement.

In another embodiment, A is-CR ₂₁R ₂₂-,-NR ₂₃-,-O-, or-S-; B is-OR ₂₄,-SR ₂₅,-NR ₂₈R ₂₉, D and E are-CR independently or jointly ₃₀-, or-N-.Preferred assorted two ring residues can be selected from:

In embodiments more of the present invention, may there be the tautomer of (1) and (2) in the compound with chemical formula (1) disclosed here, and this also belongs within the scope of the invention.

In some embodiments of the present invention, disclosed herely comprise 2, may there be (XI) in the compound of 4-thiazolidinedione, and ((XII) and tautomer (XIII), this also belongs within the scope of the invention.

When "----" when existing, shown here comprise 2, the Z of 4-thiazolidinedione compound and E configuration all belong within the scope of the invention.Z and E configuration chemical formula all may have following array structure respectively:

May comprise the salt of these compounds, for example positive ion salt at compound disclosed here.May combine the positively charged ion that forms pharmaceutical salts with The compounds of this invention and comprise basic metal, for example sodium and potassium; Alkaline-earth metal class, for example calcium; Trivalent metal class, for example aluminium.For selecting cationic unique restricted condition is to increase toxicity.Because compound has above-mentioned tautomerism, unit price, divalence or trivalent salt may depend on corresponding alkali metal.And one or more compounds shown here may be included in this compound the salt that generates with acid-responss such as nitrogen and hydrochloric acid, carboxylic acids, for example aniline of amine, aniline, replacement, pyridyl etc.Therefore, all possible salt relevant and all belong within the scope of the invention with a kind of salt that nitrogen and acid-respons generate with tautomer.

The invention provides, but be not limited to, in each example and below these specific compounds of mentioning, and they are at a kind of salt that pharmaceutically can allow:

Here

"-----" is not essential, and m is 0, and n is 1

5-[6-(2-ethylamino--7-isopropoxy-benzoxazol-5-yl)-pyrimidin-3-yl methylene radical]-thiophene  azoles-2, the 4-diketone

5-[6-(2-ethylamino--7-isopropoxy-benzoxazol-5-yl)-pyrimidin-3-yl methylene radical]-Sai azoles alkane-2, the 4-diketone

5-[6-(2-amino-7-isopropoxy-benzoxazol-5-yl)-pyrimidin-3-yl methylene radical]-thiazolidine-2, the 4-diketone

[5-(5-Diethoxymethyl-pyridin-2-yl)-7-isopropoxy-benzoxazol-2-yl]-ethyl-amine

5-[6-(2-diethylin-7-isopropoxy-benzoxazol-5-yl)-pyrimidin-3-yl methylene radical]-thiazolidine-2, the 4-diketone

5-[6-(7-isopropoxy-2-Propylamino-benzoxazol-5-yl)-pyrimidin-3-yl methylene radical]-thiazolidine-2, the 4-diketone

5-[6-(7-isopropoxy-2-Propylamino-benzoxazol-5-yl)-pyrimidin-3-yl methylene radical]-thiophene  azoles-2, the 4-diketone

5-[6-(2-ethylamino--7-isopropoxy-1 hydrogen-benzoxazol-5-yl)-pyrimidin-3-yl methylene radical]-thiophene  azoles-2, the 4-diketone

5-[6-(7-isopropoxy-2-isopropylamine base-benzoxazol-5-yl)-pyrimidin-3-yl methylene radical]-thiophene  azoles-2, the 4-diketone

5-[6-(7-isopropoxy-2-isopropylamine base-1 hydrogen-benzoxazol-5-yl)-pyrimidin-3-yl methylene radical]-thiophene  azoles-2, the 4-diketone

Make composition

Figure 13 illustrates a kind of representative solution of the synthesis path of producing these compounds disclosed here.In synthetic, a kind of chemical formula is the boric acid of (XX), and R15=H can produce biaryl (XXIV) with the aryl halide coupling that comprises a carbonyl.May induce with boron ester class such as the coupling reaction that above-mentioned biaryl (XXIV) generates, for example the R15 position combines with boron and forms a kind of 2,3-diformazan-2,3-decides the glycol boron ester (structure of boron ester: Ishiyama T.et al.J.Org.Chem.1995,60,7508-7510, Ishiyama T.et al.Tetrahedron Letters1997,38,3447-3450; Coupling pinacol esters:Firooznia, F.etal.Tetrahedron Letters 1999,40,213-216, Manickam, G.et al.Synthesis 2000,442-446; Be combined in this at these four pieces of quoted passages by quoting as proof).Biaryl (XXI) can be again through the condensing formation benzyl of 2,4 thiazolidinedioneses river (XXII).

The preparation process of a kind of chemical formula for the compound of (XXII) is provided in embodiments more of the present invention:

Wherein

A is-CR ₂₁R ₂₂-,-NR ₂₃-,-O-, or-S-;

B is-OR ₂₄,-SR ₂₅,-NR ₂₈R ₂₉

D and E distinguish or are-CR together ₃₀-, or-N-;

Ar is a chemical formula (c) or (d)

R ₁, R ₂, R ₃, R ₂₁, R ₂₃, R ₂₄, R ₂₅, R ₂₈, R ₂₉And R ₃₀Be hydrogen independently or jointly; alkyl; the alkyl that replaces; alkylhalide group; thiazolinyl; the thiazolinyl that replaces; alkynyl; the alkynyl that replaces; halogen; cyano group; nitro; hydroxyl; acyl group; the acyl group that replaces; acyloxy; amino; single substituted-amino; disubstituted amido; alkyl sulfonamide; aryl sulfonic acid amides; alkyl urea; the aryl urea; alkyl carbamate; aryl-carbamate; heteroaryl; alkoxyl group; the alkoxyl group that replaces; halogenated alkoxy; thio alkoxy; sulfo-halogen alkoxyl group; carboxyl; carbalkoxy; the alkyl carboxylic acid amides; the alkyl carboxylic acid amides that replaces; the dialkyl group carboxylic acid amides of dialkyl group carboxylic acid amides or replacement.

Comprise the following steps:

1) is connected compound that comprises the biaryl carbonyl of generation with a primary aromatic yl residue with a deputy aromatic yl residue; Wherein first aromatic yl residue comprises the residue of array structure under the having of replacement or non-replacement:

And wherein second aromatic yl residue has a carbonyl group with following array structure:

And the compound that wherein comprises the biaryl carbonyl has following (XXIV) structure:

2) based on the nitrification of (XXIV), the structural formula of condensation product is (XXV):

3) under the provide protection of ethylene glycol to carbonyl group, protected product structure formula is (XXVI):

4) under the provide protection that the palladium on the gac exists, nitryl group is reduced into amino by ammonium formiate, generates (XXVII) structure:

5) under the cyclic action of cyanogen bromide, product has the structural formula of (XXVIII):

R wherein ₁And R ₂Be hydrogen.If R ₁Be an alkyl group, above-mentioned product (XXVIII) can be carried out alkanisation by haloalkane when NaH exists.

5) after the hydrolytic action, product has the chemical formula of (XXI):

6) 2, the biaryl of condensation inclusion compound (XXI) when the 4-thiazolidinedione exists, product has the structural formula of (XXII):

The Ben Yajiaji that provides the further step reduction to have chemical formula (XXII) in another embodiment of the present invention generates the have chemical formula benzyl compounds of (XXIII):

Some are applicable to that it all is to it will be clear to those of skill in the art that these methods can be applied in the method for the present invention that reduction Ben Yajiaji compound generates the method for benzyl compounds (comprise hydrogenization, with metal hydride reactant or dissolution of metals effect).

Also may be different from above-mentioned method with other in the conversion of these used different organic groups realizes.Relevant other are used for the synthesis step of compound synthetic shown here, its reference can be at article March, J.Advanced Organic Chemistry, 4thEdition, Weiley-Interscience (1992) or Larock, R.C, Comprehensive Organic Transformations, A Guide to FunctionalGroup Preparations, VCH Publishers, Inc. find in (1989), the two all is combined in this by reference.

One embodiment of the invention relate to the process of making the compound with Formula I, generate a biaryl comprising two aromatic nucleus of coupling, have the part (moiety) of a carbonyl on one of them aryl, a preferred acetaldehyde.The biaryl product that is generated can be then at activity methene compound, and as 2, effect such as 4-thiazolidinedione generates the Ben Yajiaji compound of the chemical formula (1) of a kind of existence the "----" down by condensation.In an optional step, the Ben Yajiaji compound may be reduced and generate a kind of benzyl compounds that does not have the chemical formula (1) of "----".

Can induce two aryl rings of coupling under the effect with a kind of aryl boric acid or ester and halogenated aryl (for example iodo, bromo or chloro), trifluoromethyl yellow acid or NITRODIAZONIUM FLUOROBORATE; These are respectively at Suzuki, Pure ﹠amp; Applied Chem.66:213-222 (1994), Miyaura and Suzuki, Chem.Rev.95:2457-2483 (1995), Watanabe, Miyaura and Suzuki, Syn-lett.207-210 (1992), Littke and Fu.Angew.Chem.Int.Ed.37:3387-3388 (1998), Indolese, Tetrahedron Letters, 38:3513-3516 (1997), Firooznia, et.al.TetrahedronLetters 40:213-216 (1999), and Dorses, et.al.Butt.Soc.Chim.F7:133:1095-1102 (1996) has detailed description, all is combined in this by reference.

According to this coupling reaction, can use as (X) and the precursor (XX):

Wherein

(X) or be a trifluoromethyl sulfonic acid, a halogenide (as iodo, bromo, chloro), perhaps be a NITRODIAZONIUM FLUOROBORATE, perhaps be hydrogen, and R15 be alkyl or hydrogen.As an alternative, should be understood that these coupling groups can exchange.Above-mentioned these precursors of mentioning also can be prepared with other method by those of ordinary skills at an easy rate.For example, the boron ester can transform into corresponding lithium aryl with a kind of halogenated aryl, handles with the trialkyl borate then to prepare.Preferably, the boron ester is hydrolyzed to boric acid.This coupling reaction can also be induced with a halogenated aryl zinc and halogenated aryl or trifluoromethane sulfonic acid.Perhaps, this coupling reaction also can be realized with trialkylated tin derivative and halogenated aryl or trifluoromethane sulfonic acid.These coupling process are seen Stanforth, and Tetrahedron 6554:263-303 (1998) one literary composition summaries are combined in this by reference.Generally speaking, selecting to use which kind of special coupling reaction, generally is to consider according to existing precursor, chemo-selective, regioselectivity and three-dimensional conformation.

Use suitable activity methene compound, for example 2, the biaryl carbonyl that the 4-thiazolidinedione carries out comprise its derivative (as, Figure 13, compound (XXI)) condensation reaction, also can be finished with other method by those of ordinary skills.For example, the biaryl carbonyl product of coupling reaction can be under a kind of activity methene compound effect condensation produce a kind of chemical formula and be the benzene ylidene compounds of (I) (just when "----" when existing), as Tietze and Beifuss, Compre-hensive Organic Synthesis (Pergamon Press), 2:341-394, (1991) one literary compositions are described, are combined in this by reference.Those of ordinary skills know that the intermediate product with those scopes of oh group can produce, and is as follows in the condensation reaction of the biaryl carbonyl that comprises derivative and a kind of activity methene compound.

The oh group that these intermediate products comprise often is removed (as water) in condensation course, generate the Ben Yajiaji compound of expection.Yet, thereby the condition of reaction can be used to separate or further handles the hydroxyl that comprises intermediate product through revising, and the embodiment of this class also belongs within the scope of the invention.The generation for the intermediate product condensation reaction of compound (XXI) and the reaction of a kind of activity methene compound has been described in reaction shown in above-mentioned.The required effective catalyzer of condensation reaction can be selected from ammonium, and no matter primary amine, secondary amine and tertiary amine are free alkali or the amine salt that comprises machine acid, as acetate.The example of catalyzer comprises the pyridine of tetrahydro-arsenic, six pyridine of hydrogen arsenic, arsenic pyridine, diethylamine and relevant acetates.Organic catalyst also can be used for this condensation reaction.Inorganic catalyzer comprises, still is not limited only to, and titanium tetrachloride and a kind of tertiary base are as the arsenic pyridine; Magnesium oxide in inert solvent and zinc oxide.Such condensation reaction has very strong solvent dependency, need carry out routine test with specific catalyst as everyone knows and identify the suitableeest solvent, and solvent comprises ethanol, tetrahydrofuran (THF), two  azoles or toluene preferably, or its relevant mixture.What use among the present invention is 2, the 4-thiazolidinedione.Institute's synthetic Ben Yajiaji (as, Figure 13, compound (XXII)), as hope, reducible is a kind of compound that does not contain "----" chemical formula for (I).

The utilization of compound

In many biological test processes, comprise in vivo test and in vitro tests, have been found that compound of the present invention is effective in human relative disease or representational human diseases experimental study process.For example, the kinase whose compound of Flt-3 can be suppressed and leukaemia cancer cell can be killed and wounded.Comprise in prominent Flt3 and the KIT modification kinases at detected about 180 kinds of different kinases, find that compound 1 can specific inhibition kinase activity.According to Ambit kinases shaker test (M.A.Fabian et al.Nature Biotechnology, 23 pp.329-336, March 2005) detect and find only have six kinds of kinases to be suppressed in all kinases, it suppresses constant (or binding constant Kd) in inferior micro-molar range (see figure 1).These test-results show that said chemical structure can suppress a series of protein kinases, and have selectivity simultaneously.By detection compound to MV4; The biological effect that its selectivity suppresses Flt3 is observed in the influence of 11 cell proliferation efficient.These human AML cells (expressing the Flt-3 mutant) are divided equally in several 96 orifice plates, and each has the compound 1,2,3 or 4 of different concns.Use 3-4,5-dimethylthiazole-2,5-hexichol tetrazolium (MTT) measuring cell propagation effect is observed compound to human leukemia cancerous cell line MV4; The retarding effect of 11 cell proliferations and cell killing effect.Compound 1,2,3 and 4 can significantly suppress MV4 in dose-dependent mode; The propagation of 11 cells and kill and wound MV4; 11 cell (see figure 2)s.The specific cell Mortaility results that Fig. 3 shows shows the leukaemia cancer cell that kills and wounds that compound 1 can be special, and does not have selective killing effect for prostate cancer cell (PC-3) and pancreatic cancer cell (Bx-PC-3).Comprehensive these data are considered, the inhibition Flt-3 that compound 1 can be special is described, thereby produce the MV4 to the Flt-3 sudden change; The cell killing effect of 11 cells.Compound 2 is in the external activity that can selectivity suppresses tumour cell, and compares with compound 1 and to have more special kinase inhibition effect.Result shown in Figure 4 shows, when having detected about 180 kinds of different kinases according to Ambit kinases shaker test, only can suppress three kinds of different kinases and their some mutant in concentration for the micromole or near micromolar compound 2.Be the bioavailability of detection compound, give the compound 2 of the 10mg/kg of three rat single port clothes dosage, the time point extracting blood sample different detects its Plasma Concentration.After single port clothes dosage gave compound 2, Plasma Concentration surpassed 2 μ M in two hours, and can reach the peak concentration value (C of 3 μ M in ten hours _Max), and the peak can continue to surpass 24 hours (see figure 5)s.Because high blood level can be kept several hrs and good oral administration biaavailability, suggestion compound 2 is oral administration once a day.

Utilize subcutaneous xenotransplantation knurl model evaluation compound 2 effect in vivo.The mouse bare subcutaneous injection of extracing thymus gland is expressed the MV4 of the Flt-3 of sustained activation; 11 cells, be the leukemia model (O ' Farrell A-M that has been determined, Abrams TJ, Yuen HA, Ngail TJ, Louie SG, Yee KWH, Wong LM, Hong W, Lee LB, Town A, Smolich BD, Manning WC, Murray LJ, Heinrich MC and Cherrington JM (2003) SUl 1248 is a novel Flt-3 tyrosine kinase inhibitor with potentactivity in vitro and in vivo.Blood 101:3597-3605).Mouse bare subcutaneous injection MV4; In several weeks, be full of leukaemia cancer cell in the spleen of these mouse enlargements behind 11 cells, mouse is sick or dead usually.Gather in the crops MV4 in exponential phase of growth; 11 cells are promptly a kind ofly expressed the human leukaemia cell system that the Flt-3 internal series-connection repeats mutant nucleotide sequence, use then matrigel (BD Biosciences, Bedford, MA) resuspended.Give the nearly hind leg flank of the nude mice place injection 5 * 10 of extracing thymus gland in the time of 0 day ⁶Individual MV4; 11 cells.Come the effect of assessing compound oral administration 2 every day treatment by two kinds of different tests.

Prevent the development of the acute grain of nude mice leukemia tumour

Mouse bare subcutaneous injection MV4; 11 cells.When having grown tumour in tumour initial two all backs and animal body, one group of mouse orally give 30mg/kg compound 2 once a day, continues 8 and 14 days, and second group gives placebo treatment.In the process of treatment, use the vernier caliper measurement tumor size weekly for twice, and volume calculation is that (Tomayko MM5Reynolds CP (1989) Determination of subcutaneous tumor size inathymic nude mice.Cancer Chemother Pharmacol 24 (3): 148-54), ellipsoidal volume is by the size decision of heteroplastic Subcutaneous tumor for ellipsoidal volume.Histogram is represented every group of five animals.Test finds that oral administration 30mg/kg compound 2 can significantly suppress human leukemia cancer cells MV4 once a day; The growth (see figure 6) of 11 transplantation tumors.After handling a week again, the difference of drug treating group and control group is more and more obvious.Fig. 6 shows the drafting pattern as a result with drug treating 8 and 14 days.In a word, compound 2 can significantly suppress growth of tumor.And still have same effect during to off-test in the 24th day.

Treatment to the huge tumor set up

In the research of another series, about 5,000,000 MV4 of mouse bare subcutaneous injection; 11 cells.Be about 100mm in the tumour size after three weeks ³After, mouse is divided into two groups (6 every group).One group of orally give 50mg/kg compound 2, another group gives placebo, once a day, continues for two weeks.Then tumour can be divided into according to the size of tumour detection less than/disappear several groups of static or lasting increases.The tumour of one group of mouse of placebo treatment was grown up to 1000mm within two weeks ³, and have 66% its tumour size to change with the mouse that compound 2 (50mg/kg) was handled.Having 50% in these mouse, to demonstrate its tumour be resting state, and remaining 50% mouse tumor is eliminated (see figure 7) fully.

Compound 5 can selective killing MV4; 11 leukaemia cancer cells, and prostate cancer and pancreatic cancer cell are not had selective killing effect.For prostate cancer cell and pancreatic cancer cell, their propagation does not need the Flt-3 (see figure 8).In addition, in the kinase inhibition shaker test, compound 5 can optionally suppress Flt-3, and can effectively suppress the Flt-3 mutant of all detections.Compound 5 also can suppress the RET kinases on inferior micro-molar concentration level.As everyone knows, RET can promote the propagation (see figure 4) of thyroid carcinoma cell.In the rat body, compound 5 has excellent drug dynamic metabolism characteristic: single oral dose gives that Plasma Concentration surpasses 14 μ M in the 12mg/Kg compound 5, two hours, and the peak can continue to reach 15 hours (see figure 9)s.In nude mice model, one day twice oral 15mg/Kg compound 5 can suppress the growth (see figure 10) of human leukemia transplantation tumor.In Figure 11, by measuring the therapeutic efficiency that nude mice model individual tumors volume comes comparative compound 2 and compound 5 and placebo.Every group of 5-8 animal gives placebo or compound 2 (30mg/kg once a day) or compound 5 (one day twice of 15mg/kg) and handles, and continues medication 10 days.Two kinds of compounds and placebo are compared and can both be weakened growth of tumor.In addition, compound 5 and compound 2 mutually specific energy obviously slow down growth of tumor (seeing Figure 11).Nude mice to existing large-scale leukemia tumour gives compound 5,50mg/Kg, and processing can significantly suppress growth of tumor (seeing Figure 12).

Another aspect of the present invention relates to the using method of said compound.These compounds both can use separately also and can severally use simultaneously, and pharmaceutical composition can be used to treat mammalian diseases, especially in anthropogenetic disease.The mixture that can use said compound and comprise them by number of ways, for example, oral, enterally administering, intestines external administration, topical, snuffing are gone into, intravaginal administration, hypogloeeis or inhalation, are used for the treatment of leukemia and other cancers and comprise AML, chronic myelocytic leukemia and solid tumour.Comprehensive route of administration known in the art and dosage can be at, medicinal chemistry for example, volume S, and Hanscb, C.Pergamon Press finds in 1990, by reference it is combined in this.These compounds described herein also can be used as the disease conditioning agent of uncontrollability propagation.Be that some are representative below, but be not limited to this type of tumour: lymphoma, Hodgkin's disease, granulocyte leukemia, bladder cancer, cerebral tumor, incidence cancer, renal cancer, lung cancer, small cell lung cancer and nonsmall-cell lung cancer, myelomatosis, neuroglia blastoma, ovarian cancer, carcinoma of the pancreas, prostate cancer, skin carcinoma, liver cancer, melanoma, large bowel cancer, cervical cancer, breast cancer, and epithelial cancer.

Though said compound can be used as the administration of pure chemistry product, preferred active ingredient administration with medicinal mixture.Therefore, another aspect of the present invention should relate to one or more compounds of the present invention and combine with one or more pharmaceutically admissible associated salts, and no longer combine with the related vector that pharmaceutically allows more, and randomly with the use of other treatment and/or preventative composition.Should must there be too big harm with other constituents compatibilities and to relevant user in a sense by (these) carrier.

Medicinal mixture comprises that those are fit to that oral, enterally administering, intestines external administration (comprising intramuscular injection, subcutaneous and intravenous injection), topical, snuffing are gone into, the constituent of intravaginal administration, hypogloeeis or inhalation.This pharmaceutical composition may be with more convenient separate dosage units form or with additive method preparation general in the pharmaceutical industry under suitable situation.These methods comprise activated compound and liquid vehicle, solid substrate, semi-solid carrier, micro-solid carrier or combination are combined, and then as need, product can be moulded the form for being complementary with medication delivery system.The pharmaceutical composition that is fit to oral administration route can be made independently dose unit, as hard soft capsule, and cachet or tablet, every comprises quantitative in advance activeconstituents; Also can do powdered or particle; Perhaps make a kind of solution, suspension or emulsion.Activeconstituents also can be made pill, electuary or paste.Oral tablet and capsule have conventional vehicle, as wedding agent, weighting agent, lubricant, disintegrating agent or humectant.Tablet can add coat according to the method for bibliographical information, for example, and the enteron aisle dressing.

Oral liquid formulations can be made as water-based or oily suspensions, solution, emulsifying agent, syrup or elixir or a kind of exsiccant product, and the time spent takes with water or other suitable carriers.This liquid preparation can comprise conventional additive, as suspension agent, emulsifying agent, on-liquid carrier (comprising edible oil), or one or more sanitass.

This compound also can be mixed with and (for example be used for the intestines external administration, injection be interrupted to be injected or persistent instillation) formulation, also can be prepared into unit dosage form, place in ampoule, pre-filled syringe, low dose of infusion vessel or the multi-dose container, and add sanitas.The form that composition can adopt is that suspension, solution or emulsion as oiliness or aqueous carrier also can comprise forming agent such as suspension agent, stablizer and/or dispersion agent.Perhaps, activeconstituents also can exist by powder type, and powder obtains by aseptic separation or the solution distillation drying to sterile solid, and combines with suitable carriers, and for example powder was dissolved in aseptic apyrogenic water earlier before usefulness.If compound uses through contact skin, compound should be made ointment, emulsifiable paste or lotion, or make the patch that comprises activeconstituents.The transdermal drug delivery system that is fit to has had bibliographical information, as people such as Fisher (U.S.Pat.No.4,788,603, be combined at this by reference) or Bawas et al. (U.S.Pat.Nos.4,931,279,4,668,504 and 4,713,224, be combined at this by reference).Ointment and emulsifiable paste for example can increase or suitably thicken and/or the jelling agent moulding with water-based or oleaginous base.Lotion can also can comprise one or more emulsifying agent, stablizer, dispersion agent, suspension agent, thickening material or tinting material moulding with water-based or oleaginous base.Activeconstituents also can pass through the electrophoresis administration, and document is just like U.S.Pat.Nos.4, and 140,122, or 4,383,529,4,051,842, be combined at this by reference.The pharmaceutical dosage form that is fit to oral topical therapeutic comprises that unit dosage form as the lozenge that comprises activeconstituents usually and the shaping that combines of sucrose and dulcet matrix such as gum arabic or Tragacanth; The lozenge that comprises activeconstituents generally is carrier with the inert base, as gelatin and glycerine or sucrose and gum arabic; The mucous membrane that comprises activeconstituents sticks together glue and mouthwashes can be with suitable liquid vehicle.If necessary, mixture described above can be prepared as the form of used activeconstituents being kept release, for example, by with the combination of possess hydrophilic property polymeric matrix, and for example, with natural gel, synthetic polymer gel or mixture combination.Pharmaceutical composition of the present invention also may comprise other adjuvants such as seasonings, pigment, biocide or sanitas.

In this and otherwise preferred embodiment of invention, vehicle is the part of its composition, at least this compound that comprises a kind of carrier and some amount is enough to reach desired therapeutic effect, wherein compound to have the people of a subgroup to try out at least and reach desired therapeutic effect (as, " effective dose " of certain drug or other active agents depends on the circumstances).At this, " treatment effective dose " refers to that object of this pharmacological agent of needs takes in the certain amount in the time of can reaching effective result of treatment.In cancer treatment procedure, " dose therapeutically effective " refers to that the dosage of absorption for evaluated people can produce considerable effect.Identify a kind of medicine effective dose that contains mixture of the present invention, can finish easily by those of ordinary skills.

The dosage of the dosage of the compound of required use or active salt or derivatives thereof in the therapeutic process, not only relevant with the specific salt of selecting, and relevant, and at last by supervision doctor or clinician's decision with medicine absorption approach, treatment condition, age and patient's state.Generally speaking, those of ordinary skills can go out another kind of Mammals such as people's possible data according to the in vivo test inferred from input data that obtains on model animals such as nude mice.These deductions are not only according to what the body weight of two kinds of biologies was inferred metabolism difference, drug absorption and the route of administration of also considering them and are differently made deduction.Consider many this type of factors, proper dosage is normally between about 0.5--100mg/kg/ days, and the about 75mg/kg/ of 1--days, 3--50mg/Kg/ days or 6--90mg/kg/ days.Compound for example, has 0.5--5000mg with unit dosage form administration easily, and 5--750mg is per unit dosage form activeconstituents 10--500mg more easily.

One of skill in the art will recognize that these typical ranges outer dosage and formulation can be detected, in due course, also can be used for method of the present invention.In some specific embodiments, the Plasma Concentration peak that the active constituents of medicine of absorption reaches is about the M from 0.5--75 μ, from 1--50 μ M, or from 2--30 μ M.Can also can instil the perhaps oral pill that contains the about 0.5--500mg of activeconstituents by the active ingredient solution of intravenous injection 0.05--5% with salt solution.Can reach the blood level of expection by the activeconstituents of continuous injection infused drug 0.01--5.0mg/kg/ hour or the about 0.4--15mg/kg of discontinuity input.Projected dose can reach by dual mode, and first kind is the single dose single administration, and another kind is to be divided into several dosage with the proper spacing administration, as one day administered twice, and three times, four times or more times.Divided dose itself might further be divided, for example by different administrations, as sucking several times from sucker or liquid medicine being splashed into eyes.

Term " treatment " or " execute and control " are meant any methods of treatment at a kind of disease or obstacle, comprise prevention or disease preventing and treating or obstacle (that is: clinical symptom no longer being developed); Suppress disease or obstacle (for example suppress the development of clinical symptom and/or palliate a disease or illness, alleviate clinical symptom).Be appreciated that because final inducement or this type of factor the unknown or latent, so be not always well to distinguish " prevention " and " inhibition " a kind of disease or obstacle.Correspondingly, term " protection " should be understood that " treatment " of forming a kind of form realizes " prevention " and " inhibition ".Therefore " protection " speech has also comprised " prevention ".

Example

Following goods and example are used to make those of ordinary skills more clearly to be familiar with and put into practice the present invention.They should not be regarded as having limited scope of the present invention, and are relevant explanation and the representative as it.

Example one: representative compounds and theirs synthetic

Compound 1

Compound 2

Compound 3

Compound 4

Compound 5

Compound 6

Compound 7

Compound 8

Synthesizing of compound 2

To 6-(2-ethylamino-7-isopropoxy-benzene  azoles-5-yl)-pyrimidine-3-formaldehyde (0.385g, 1.183mmol) add piperidines (23 μ l in the solution in 2ml toluene, 0.237mmol) and acetate (23 μ l, 0.237mmol, add thiazolidine-2,4-diketone (0.139g, 1.183mmol) solution in 2ml toluene.Reaction mixture is heated to reach and refluxed four hours.The product of separating out after filtration, dry back output 0.395g 5-[6-(2-ethylamino-7-isopropoxy-benzene  azoles-5-yl)-pyrimidine-3-methylene radical]-thiazolidine-2,4-diketone yellow solid (78.7% output capacity); HPLC analyzing and testing purity is 99.59% (355nm); 99.40% (225nm); 280 ℃-281 ℃ of mp. ¹H?NMR(500MHz，DMSO-d ₆)：20t，J＝7.26Hz，3H)；1.34(d，J＝6.04Hz，6H)；3.35(m，2H)；4.87(m，1H)；7.54(d，J＝1.24Hz，1H)；7.64(d，J＝1.17Hz，1H)；7.86(s，1H)；7.97(dd，J1＝2.39Hz，J2＝8.52Hz，1H)；8.03(t，J＝5.63Hz，1H)；8.14(d，J＝8.50Hz，1H)；8.88(d，J＝2.30Hz，1H)；12.7(br?s，1H)。

(2-ethylamino-7-isopropoxy-benzene  azoles-5-yl)-pyrimidine-3-prepared formaldehyde is as follows for this intermediate product 6-:

6-(2-amino-7-isopropoxy-benzene  azoles-5-yl)-pyrimidine-3-formaldehyde (0.860g, 2.893mmol) and salt of wormwood (0.960g, 6.942mmol) mixture in 20ml DMF at room temperature stirred one and a half hours.Add then iodoethane (278 μ l, 3.472mmol).Reaction was at room temperature stirred three days.Dilute this solution with the 20ml vinyl acetic monomer, successively water (2 * 30ml) and salt solution (30ml) washing, use anhydrous magnesium sulfate drying, filter then and evaporate.Residue carries out stratographic analysis by the silicon gel and handles (elutriant: output 0.385g 6-2-ethylamino-7-isopropoxy-benzene  azoles-5-base-pyrimidine-3-formaldehyde yellow solid (41% output capacity) 100% vinyl acetic monomer). ¹H?NMR(500MHz；DMSO-d ₆)：1.98(t，J＝7.31Hz，3H)；1.35(d，J＝6.09Hz，6H)；3.36(m，2H)；4.88(m，1H)；7.57(d，J＝1.35Hz，1H)；7.69(d，J＝1.4Hz，1H)；8.01(t，J＝5.6Hz，1H)；8.21(d，J＝8.35Hz，1H)；8.26(dd，J1＝2.2Hz，J2＝8.35Hz，1H)；9.12(d，J＝1.65Hz，1H)；10.12(s，1H)。

(2-amino-7-isopropoxy-benzene  azoles-5-yl)-pyrimidine-3-prepared formaldehyde is as follows for this intermediate product 6-:

(1.00g 2.69mmol) stirs and is warmed up to 80 ℃ with the mixture of 0.5N hydrochloric acid and kept 30 minutes 5-(5-diethoxy methyl-pyrimidine-2-base)-7-isopropoxy-benzene  azoles-2-azanol.The product of preparation water filtration and washing.Filtrate is regulated pH to 7 with saturated sodium bicarbonate, and (3 * 15ml) extract with vinyl acetic monomer.Organic phase is water (50ml) and salt solution (50ml) washing successively, uses anhydrous magnesium sulfate drying, filters evaporation.Residue and cross filtered product and combine dry back output 0.860g 6-2-amino-7-isopropoxy-benzene  azoles-5-base-pyrimidine-3-formaldehyde (quantitatively output). ¹H?NMR(500MHz；DMSO-d ₆)：1.35(d，J＝6.00Hz，6H)；4.90(m，1H)；7.59(d，J＝1.44Hz，1H)；7.66(d，J＝1.5Hz，1H)；7.78(br?s，2H)；8.25(m，2H)；9.12(m，1H)；10.12(s，1H)。

Being prepared as follows of intermediate product 5-(5-diethoxy methyl-pyrimidine-2-base)-7-isopropoxy-benzene  azoles-2-azanol:

2-amino-4-(5-[1,3] dioxolane-2-base-pyrimidine-2-base)-6-isopropoxy-phenol (1.273g, 4.02mmol) dropwise add in the mixture in dehydrated alcohol (60ml) the 5M cyanogen bromide acetonitrile solution (2.41ml, 12.07mmol).The mixture that stirring generates also refluxed 1.5 hours in argon gas.The reaction mixture of finishing under reduced pressure concentrates, and dilutes with vinyl acetic monomer (50ml) and water (20ml).Water can be used vinyl acetic monomer, and (2 * 20ml) separate and extraction.The pH value of regulating water with saturated sodium bicarbonate is about 7, and extracts with vinyl acetic monomer (15ml).Organic phase is water (50ml) and salt solution (50ml) washing successively, uses anhydrous magnesium sulfate drying, filters evaporation.Residue obtains 1.118g 5-(5-diethoxy methyl-pyrimidine-2-base)-7-isopropoxy-benzene  azoles-2-azanol (81% output capacity) by silicon gel chromatography analyzing and processing (elutriant: vinyl acetic monomer/hexane, 4: 1). ¹H?NMR(500MHz；DMSO-d ₆)：15(t，J＝7.01Hz，6H)；1.31(d，J＝6.03，Hz，6H)；3.50(m，2H)；3.57(m，2H)；4.82(m，1H)；5.58(s，1H)；7.41(m，1H)；7.43(s，2H)；7.76(dd，J1＝2.20Hz，J2＝8.30Hz，1H)；7.93(m，1H)；8.59(d，J＝2.18Hz，1H)。

Being prepared as follows of intermediate product 2-amino-4-(5-[1,3] dioxolane-2-base-pyrimidine-2-base)-6-isopropoxy-phenol:

To 4-(5-[1,3] dioxolane-2-base-pyrimidine-2-base)-2-isopropoxy-6-nitro-phenol (1.596g, 4.96mmol) (1.45g 23.0mmol), is the palladium that is added in 10% weight on the gac (0.23g) subsequently to add ammonium formiate in the mixture in dehydrated alcohol (80ml).The mixture that stirring is generated also made it to reflux 1.25 hours under argon gas.To react the back miscellany and be cooled to 23 ℃ and filter the silicon grain.Permeate concentrates and uses successively vinyl acetic monomer (150ml) and water (50ml) washing under depressurization.Water can be used vinyl acetic monomer, and (3 * 15ml) separate and extraction.Organic phase is water (50ml) and salt solution (50ml) washing successively, uses anhydrous magnesium sulfate drying, filters evaporation.Residue obtains 1.273g 2-amino-4-(5-[1,3] dioxolane-2-base-pyrimidine-2-base)-6-isopropoxy-phenol (87% output capacity) by silicon gel chromatography analyzing and processing (elutriant: vinyl acetic monomer/hexane, 3: 2). ¹H?NMR(500MHz；DMSO-d ₆)：1.47(d，J＝6.12Hz，6H)；4.80(m，1H)；7.91(d，J＝8.3Hz，1H)；8.09(d，J＝2.0Hz，1H)；8.25(dd，J1＝2.05Hz，J2＝8.3Hz，1H)；8.40(d，J＝2.1Hz，1H)；9.11(d，J＝1.90Hz，1H)；10.08(s，1H)；10.86(s，1H)。

Being prepared as follows of intermediate product 4-(5-[1,3] dioxolane-2-base-pyrimidine-2-base)-2-isopropoxy-6-nitro-phenol:

To 6-(4-hydroxyl-3-isopropoxy-5-nitro-benzene)-pyrimidine-3-formaldehyde (1.507g, 4.99mmol) mixture in dry toluene (30ml) add successively ethylene glycol (5.6ml, 99.7mmol) and the toluenesulphonic acids monohydrate (57mg, 0.3mmol).Stir the mixture that is generated then and under argon gas, make it to reflux 5 hours with Dean-Stark trap.The reaction mixture of finishing is cooled to 23 ℃, and also regulating its pH value with 10% solution of potassium carbonate is about 7, uses the dilution of vinyl acetic monomer (30ml) and water (20ml) then successively.Water can be used vinyl acetic monomer, and (3 * 15ml) separate and extraction.Organic phase is water (30ml) and salt solution (30ml) washing successively, uses anhydrous magnesium sulfate drying, filters evaporation.Residue obtains 1.596g 4-(5-[1,3] dioxolane-2-base-pyrimidine-2-base)-2-isopropoxy-6-nitro-phenol (92% output capacity) by silicon gel chromatography analyzing and processing (elutriant: vinyl acetic monomer/hexane, 3: 7). ¹H?NMR(500MHz；DMSO-d ₆)：1.41(d，J＝6.04Hz，6H)；4.79(m，1H)；6.15(br?s，1H)；7.02(d，J＝8.35Hz，1H)；7.55(dd，J1＝2.0Hz，J2＝8.35Hz，1H)；7.79(m，2H)；8.16(dd，J1＝2.10Hz，J2＝8.35Hz，1H)；9.04(d，J＝2.15Hz，1H)；10.08(s，1H)。

Being prepared as follows of intermediate product 6-(4-hydroxyl-3-isopropoxy-5-nitro-phenyl)-pyrimidine-3-formaldehyde:

To 6-(4-hydroxyl-3-isopropoxy-phenyl)-pyrimidine-3-formaldehyde (2.864g, 11.13mmol) in the mixture in trifluoroacetic acid (30ml) 0 ℃ add down saltpetre (1.18g, 11.69mmol).Under 0 ℃ of ar gas environment, the mixture that is generated was stirred 45 minutes then.The reaction mixture of finishing was poured on ice and stirs three hours.Extract solution with vinyl acetic monomer solvent (30ml).Water is neutralized to the pH value with solid sodium bicarbonate is about 7, and use ethyl acetate extraction once more.Organic phase is water (30ml) and salt solution (30ml) washing successively, uses anhydrous magnesium sulfate drying, filters and evaporation.Residue obtains 2.621g 6-4-hydroxyl-3-isopropoxy-5-nitro-phenol-pyrimidine-3-formaldehyde (78% output capacity) by silicon gel chromatography analyzing and processing (elutriant: vinyl acetic monomer/hexane, 2: 3). ¹H?NMR(500MHz；DMSO-d ₆)：1.47(d，J＝6.12Hz，6H)；4.80(m，1H)；7.91(d，J＝8.3Hz，1H)；8.09(d，J＝2.0Hz，1H)；8.25(dd，J1＝2.05Hz，J2＝8.3Hz，1H)；8.40(d，J＝2.1Hz，1H)；9.11(d，J＝1.90Hz，1H)；10.08(s，1H)；10.86(s，1H)。

Being prepared as follows of intermediate product 6-4-hydroxyl-3-isopropoxy-phenyl-pyrimidine-3-formaldehyde:

To 6-[4-(tertiary butyl-dimethyl-siloxy-)-3-isopropoxy-phenyl]-pyrimidine-3-formaldehyde (3.0g, 8.07mmol) be dissolved in mixture in the anhydrous tetrahydro furan (60ml) and dropwise add tetrabutylammonium fluoride (1.0M is dissolved in anhydrous tetrahydro furan, 9.69ml, 9.69mmol).Under 23 ℃ of ar gas environments, the mixture that is generated was stirred 1.5 hours then, use vinyl acetic monomer (30ml) and water (30ml) to dilute this solution then successively.Water can be used vinyl acetic monomer, and (2 * 20ml) separate and extraction.The organic phase that merges is water (15ml) successively, 0.1N hydrochloric acid (22ml) and salt solution (15ml) washing.The organic phase anhydrous magnesium sulfate drying filters, evaporation.Residue obtains 1.90g 6-4-hydroxyl-3-isopropoxy-phenyl-pyrimidine-3-formaldehyde (91% output capacity) by silicon gel chromatography analyzing and processing (elutriant: vinyl acetic monomer/hexane, 1: 4). ¹H?NMR(500MHz；DMSO-d ₆)：1.41(d，J＝6.04Hz，6H)；4.79(m，1H)；6.15(br?s，1H)；7.02(d，J＝8.35Hz，1H)；7.55(dd，J1＝2.0Hz，J2＝8.35Hz，1H)；7.79(m，2H)；8.16(dd，J1＝2.10Hz，J2＝8.35Hz，1H)；9.04(d，J＝2.15Hz，1H)；10.08(s，1H)。

Intermediate product 6-[4-(tertiary butyl-dimethyl-siloxy-)-3-isopropoxy-phenyl]-being prepared as follows of pyrimidine-3-formaldehyde:

With 6-bromo-pyrimidine-3-formaldehyde (4.70g, 25.2mmol), [4-(tertiary butyl-dimethyl-siloxy-)-3-isopropoxy-phenyl]-boric acid (11.72g, 37.80mmol) and salt of wormwood (6.97g, 50.4mmol) mixture in toluene (100ml), ethanol (17ml) and water (15ml) at room temperature outgased 20 minutes with argon gas.(1.46g 1.26mmol), makes mixture be issued at argon gas then and refluxes and stirred 16 hours at argon gas to add tetrakis triphenylphosphine palladium (0).With the mixture solution cool to room temperature, with vinyl acetic monomer (75ml) diluting soln, anhydrous magnesium sulfate drying is used in water (40ml) and salt solution (40ml) washing successively then, filters and evaporation.Residue obtains 6-[4-(tertiary butyl-dimethyl-siloxy-)-3-isopropoxy-phenyl of 11.403g by silicon gel chromatography analyzing and processing (elutriant: vinyl acetic monomer/hexane, 1: 4)]-pyrimidine-3-formaldehyde (95% output capacity).

Being prepared as follows of intermediate product [4-(tertiary butyl-dimethyl-siloxy-)-3-isopropoxy-phenyl]-boric acid:

To put into 2 liter of three neck round-bottomed bottle that air driven agitator is arranged at the top at the n-Butyl Lithium of the 2.5M of the 80ml in the hexane (0.33mol), and place dry ice acetone bath to be cooled to-78 ℃ solution.Under argon gas, dropwise add in this solution of-78 ℃ and comprise (4-bromo-2-isopropoxy-phenoxy group)-tertiary butyl-dimethyl-silicomethane (76g, tetrahydrofuran solution 0.22mol) (60ml).Under 0 ℃, reaction mixture solution was stirred 1 hour, dropwise add then triisopropyl borate ester (152ml, 0.660mol).0 ℃ of following stirred reaction mixture solution 2 hours.Slowly add saturated aqueous solution ammonium chloride (80ml) in the reaction mixture.Extract the mixture solution that quenches with vinyl acetic monomer solvent (200ml).Organic layer is water (150ml) and salt solution (150ml) washing successively, then through anhydrous magnesium sulfate drying.After filtering out sal epsom, evaporated filtrate.Residue obtains [4-(tertiary butyl-dimethyl-siloxy-)-3-isopropoxy-phenyl]-boric acid (56% output capacity) of 38.5g by silicon gel chromatography analyzing and processing (elutriant: vinyl acetic monomer/hexane, 5: 95).

Being prepared as follows of intermediate product (4-bromo-2-isopropoxy-phenoxy group)-tertiary butyl-dimethyl-silicomethane:

With the tertiary butyl-dimethyl chlorination silicomethane (21.4g, 0.142mol), 4-nitrogen-dimethylamine pyrimidine (0.461g, 0.004mol) and triethylamine (19.8ml 0.142mol) joins and comprises 4-bromo-2-isopropoxy-phenol (23.5g is in 160ml dimethyl formamide solution 0.102mol).The mixture of being produced at room temperature stirred 17 hours.Use vinyl acetic monomer (200ml) diluted reaction mixture and water (150ml) and salt solution (150ml) washing successively then, through anhydrous magnesium sulfate drying.After filtering out sal epsom, evaporated filtrate.Residue by silicon gel chromatography analyzing and processing (elutriant: hexane) obtain (4-bromo-2-isopropoxy-phenoxy group)-tertiary butyl-dimethyl-silicomethane of 32.3g.

Being prepared as follows of intermediate product 4-bromo-2-isopropoxy-phenol:

The tribromide pyridine (116g, 0.362mol) in the suspension of 250ml methylene dichloride, be added in 2-isopropoxy phenol in the 150ml methylene dichloride (50g, 0.329mol).Under the room temperature mixture was stirred 6 hours.(1N 200ml) makes its quenching to add aqueous hydrochloric acid in the reaction mixture.After the separation, organic phase is washed through saturated sodium thiosulfate (150ml) and salt solution (150ml) successively, and uses anhydrous magnesium sulfate drying.After filtering out sal epsom, evaporated filtrate is with 4-bromo-2-isopropoxy-phenol (94% productive rate) of output 71g.

Example two: the analysis of kinase inhibition characteristic profile

As preliminary screening, the compound of 10 μ M is tested at one group of 180 kinds of kinases.Modal Flt-3 and cKit mutant have been comprised in these kinases.(method of describing in A small molecule-kinase interaction map for clinical kinaseinhibitors.Nature Biotechnology 23:329-36 (2005) article does the kinase assay test and binding constant detects according to people such as Fabian.In brief, human kinase is with T7 phage fusion protein form expression, and a little cover fixed probe part combines with free test compound.If the combination of free test compound is also sealed the Triphosaden site, can be incorporated into the part that is fixed on the solid support with regard to few of protein molecular so.The result can read by the fusion rotein amount of calculations incorporated on solid support.For each kinases, compound is done hit or " not hitting " by initial note in the preliminary screening process.Being labeled as the compound that hits has a quantitative score, and is called as " control per-cent ".The result that Fig. 1 concludes shows that compound 1 can suppress several kinases, and

compound

2,5 and 8 has significant specificity, can only be specifically in conjunction with the kinases target of only a few.

To compound 2,3,5 and 8 have carried out second trial, and (the described method of Asmall molecule-kinase interaction map for clinical kinaseinhibitors.Nature Biotechnology 23:329-36 (2005) determines that compound is to specific kinase whose binding constant (Kd) according to people such as Fabian.The result shows that compound 1 is a kind of strong inhibitor of Flt-3 (Kd 100-200nM), and have wide kinase inhibition characteristic profile, as suppressing AbI and PDGFRD, FGFR3, CSF1R, RET, Kit, JAK2 and Aurka kinases (Fig. 1) to 714nM at 80nM.Therefore, compound 1 might be as the introducing structure of the kinase inhibitor of the more highly selective that is used for a bigger series.Some specific residues can generate the very compound 2,5 and 8 of high specific on change compound 1 structure, and these compounds are the kinase inhibitor with high degree of specificity, only can combine with the kinases target specificity of only a few.Showed among Fig. 4 that these compounds suppress the comparison of the ability of Flt-3, KIT, PDGFRD and RET.Inhibition Flt-3 that compound 5 is not only special and important Flt-3 mutant (Kd 333-975nM), and suppress RET (binding constant 770nM), and compound 8 special inhibition Flt-3, KIT and PDGFR β (Kd 95 and 740nM between).It is highly important that at this and should be noted that compound 8 also can suppress the two mutant V559D/T670I of KIT effectively.This mutant tolerates for used clinically cKit inhibitor Gleevec, so compound 8 can be used for treating the chemical sproof gi tract mesenchymal neoplasm of Gleevec patient.

Example three: cell inhibitory effect test

At human AML clone MV4; Detected the biologically of Flt-3 inhibition cell proliferation in 11.This clone is carried the most frequent human Flt-3 sudden change.By comparative compound to MV4; The influencing effect and prostate cancer cell line (PC-3) and pancreatic cancer cell system (Bx-PC3) breeding influenced effect, the selectivity of coming the cell killing of detection compound of 11 clones.Use 3-4,5-dimethylthiazole-2, the propagation of cell is determined in 5-hexichol tetrazolium (MTT) test.In brief, cell is divided equally in 96 orifice plates.With RPMI culture medium culturing MV4; 11 cells comprise 4500mg/L glucose in the substratum; The 4mM L-glutaminate; 10U/ml penicillin; 10mcg/ml Streptomycin sulphate and 20% foetal calf serum (FBS).PC-3 and Bx-PC3 cultivate with RPMI substratum 1640, comprise the 2mM L-glutaminate in the substratum; 10U/ml penicillin; 10mg/ml Streptomycin sulphate and 10% foetal calf serum (FBS).Cell inoculation is in 96 hole tissue culturing plates, and density is about 300-500 cells/well, places 6%CO ₂In 37 ℃ of environment.Cell was handled three days with kinase inhibitor.Detect the survival rate of cell then with colorimetric analysis.This detection method can be cracked into this characteristic of purple De Jia Za crystalline based on yellow tetrazolium salts MTT under the desaturase effect in activated plastosome.Therefore, this material change occur over just have complete/have in the mitochondrial viable cell of function.Form De Jia Za dissolution of crystals and produce colored solutions, this colored solutions can carry out quantitatively at the 595nm place with the porous scanning spectrophotometer.In brief, add the 5mg/ml MTT dyestuff of 10 μ l in every hole, hatched 4 hours, add 100 μ l/ hole solubilising solution termination reactions then, solubilising solution comprises 10% sodium laurylsulfonate (SDS) and 10mM hydrochloric acid.

Compound 1 to 5 can strong inhibition cell proliferation, and kill and wound MV4 in dose-dependent mode; 11 cells, and compound 1 can not suppress the propagation (Fig. 2 and 3) of PC-3 and Bx-PC3 cancerous cell line.

Example four: pharmacokinetics is measured

Be the detection of biological availability, give the 10mg/Kg compound 2 of three single oral dosages of rat.Get the analysis of blood levels of drugs at different time point (1 hour, 2 hours, 4 hours, 7 hours, 10 hours, 12 hours and 24 hours).Compound 2 demonstrates good bioavailability.The Plasma Concentration that the administration of the compound 2 of single dose 10mg/Kg produced in two hours surpasses 2 μ M, and reaches 3 μ M peak concentration value (C in ten hours _Max) and the peak can continue above 24 hours.Can keep the high blood level of several hrs and well oral administration biaavailability prompting compound 2 can one day oral once (Fig. 5).

The compound 5 that single dose gives 12mg/Kg produced the Plasma Concentration that surpasses 10 μ M and reached 14 μ M peak concentration value (C in three hour in one hour _Max) and the peak can continue above 10 hours.In a word, aspect bioavailability, compound 5 shows than compound 2 high about 3 to 4 times (Fig. 9).

Example five: the oral administration for the treatment of compound 2 and compound 5 in the AML tumour in a kind of animal model

Utilize subcutaneous xenotransplantation knurl model to assess the vivo effect of compound 2 and compound 5.Extract the mouse bare subcutaneous injection MV4 of thymus gland; 11 cells, express the Flt-3 of sustained activation, as leukemic model (O ' Farrell et al.SUl 1248 is a novel Flt-3 tyrosinekinase inhibitor with potent activity in vitro and in vivo.Blood 101:3597-3605 (2003)).Gather in the crops MV4 in exponential phase of growth; 11 cells (a kind of human leukaemia cell system, it expresses modal Flt-3-ITD mutant), use then matrigel (BD Biosciences, Bedford, MA) resuspended.The nude mice of extracing thymus gland in the time of 0 day subcutaneous nearly hind leg flank inject 5,000,000 MV4; 11 cells.For the result of treatment of compound 2 and compound oral administration 5 every day, can acute myeloblastic leukemia and treatment take place by the nude mice that thymus gland are extractd in 5 preventions of compound 2 and compound and extract the effect of the huge tumor that the nude mice of thymus gland is pre-existing in and estimate.Generally, mouse bare subcutaneous injection 5,000,000 MV4; 11 cells.Tumor cell injection is after two weeks, and the compound 2 of one group of mouse orally give every day 30mg/kg continues 8 and 14 days (Fig. 6).In the process of treatment, use the vernier caliper measurement tumor size twice weekly, and volume calculation is ellipsoidal volume (Tomayko MM andReynolds CP 1989 Determination of subcutaneous tumor size inathymic nude mice.Cancer Chemother Pharmacol 124:148-54).Post is represented 5/group.Compound 2 can significantly suppress the growth of the heterograft of Flt-3-ITD sudden change during for 30mg/kg in concentration.

In another research series, mouse bare subcutaneous injection 5,000,000 MV4; 11 cells.Reach in tumour and to be about 100mm ³Size after, mouse is divided into two groups (6 every groups).One group gives 50mg/kg compound 2, and another group gives placebo, once a day, continues for two weeks.Tumour can be divided into according to the tumour size in the placebo treatment group detection less than/disappear several groups of static or lasting increases (Fig. 7).The tumour of placebo treatment mouse was grown up to 1000mm within two weeks ³, and have 66% on the tumour size, to demonstrate effect in the mouse of handling with MC-2002 (50mg/kg).In these mouse that respond, have 50% to show that its tumour is static, and remaining 50% mouse tumor is eliminated (data do not provide) fully.

Injection MV4; The drug compound 5 of nude mice twice orally give 15mg/kg every day of 11 cells.Tumour generates beginning back the 8th day, is divided into two groups (every group of 7-9 only) according to the big young pathbreaker mouse of tumour, gives the compound 5 of vehicle or 15mg/kg respectively.In the beginning of treatment, measure the size of tumour after five days and ten days.The mouse that gives the compound 5 of 15mg/kg shows the growth (Figure 10) of inhibition heterograft acute myeloblastic leukemia clearly.

Carry out another and tested the effect of comparative compound 2 and compound 5.Tumour generates beginning back the 18 day, is divided into three groups according to the big young pathbreaker mouse of tumour.Use vehicle respectively, the compound 5 (twice of every day) of the compound 2 (once a day) of 30mg/kg or 15mg/kg is handled, oral administration 10 days.Measure tumor size, to every group single gross tumor volume drawing (only every group of n=5-8).Give group tumour that vehicle handles long very fast.Compound 2 and compound 5 can both significantly suppress tumor growth.Reducing aspect the tumour size, compound 5 obviously is better than compound 2 (Figure 11).

In another test, the injection tumour cell is after three weeks, and the mouse that forms huge AML tumour is divided into two groups according to the tumour size.Then respectively with compound 5 (twice of the every day) oral administration 3 of vehicle or 50mg/kg and 6 days.Each single tumor size is drawn.Compound 5 shows significant inhibition (Figure 12) to big tumor growth.

All patents, patent application and the publication of mentioning in this manual all is the level of those of ordinary skill in the expression industry related to the present invention.By reference it is combined in this in full hereby at this each patent of quoting, right application and publication, no matter in specific application, whether points out clearly to quote and its combination.

Although the present invention has got in touch its specific embodiment and has been illustrated, be to be understood that it can further revise, and the application is intended to comprise of the present invention any variant, the purposes of following principle of the present invention generally or adjusts, and be included in departing from the known or habitual way scope in the field that the present invention is correlated with for this explanation, these are applicable to the above essential feature that provides, and meet the scope of claims.

Claims

1. the compound of following chemical formula or its isomers, meta-bolites, many types of variant, prodrug or their salt:

Wherein:

"----" exists or do not exist;

W is (a) or (b)

(a)

A is-CR ₂₁R ₂₂-,-NR ₂₃-,-O-or-S-;

B is-OR ₂₄,-SR ₂₅,-NR ₂₈R ₂₉

D and E are-CR jointly or independently ₃₀-or-N-;

Ar is (c) or (d)

And

2. the compound of claim 1, wherein R ₁And R ₂Be independently or jointly: the dialkyl group carboxylic acid amides or the halogenated alkoxy of the alkyl carboxylic acid amides of the alkoxyl group of the alkynyl of the thiazolinyl of the alkyl of hydrogen, alkyl, replacement, haloalkyl, thiazolinyl, replacement, alkynyl, replacement, alkoxyl group, replacement, acyl group, single substituted-amino, disubstituted amido, carboxyl, carbalkoxy, alkyl carboxylic acid amides, replacement, dialkyl group carboxylic acid amides, replacement.

3. the compound of claim 2, wherein R ₁And R ₂Be independently: the aralkyl of the alkyl of hydrogen, alkyl, replacement, aralkyl, replacement, disubstituted amido group or contain the branched alkyl group of 2 to 10 carbon atoms.

4. the compound of claim 1, wherein W is:

5. the compound of claim 4, wherein R ₃Be hydrogen, and there is key in "----" expression

6. the compound of claim 1, wherein W is

7. the compound of claim 6, wherein R ₁₁And R ₁₂Be selected from the group that following group constitutes independently or jointly: methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, sec-butyl, the tertiary butyl, amyl group, tert-pentyl and n-pentyl.

8. the compound of claim 1, wherein R ₁And R ₂Be hydrogen or alkyl independently or jointly.

9. the compound of claim 1, wherein Ar is a chemical formula (c) or (d):

10. the compound of claim 2, wherein R ₂Be hydrogen.

11. the compound of claim 2, wherein there is key in "----" expression.

12. the compound of claim 2, wherein R ₃Be hydrogen, methyl or ethyl.

13. the compound of chemical formula (II) or its isomers, meta-bolites, many types of variant, prodrug or their salt:

Wherein:

R ₁Be hydrogen; And

R ₂And R ₂₄Be independently of one another or jointly: alkyl; the alkyl that replaces; haloalkyl; thiazolinyl; the thiazolinyl that replaces; alkynyl; the alkynyl that replaces; acyl group; the acyl group that replaces; acyloxy; amino; single substituted-amino; disubstituted amido; alkyl sulfonamide; aryl sulfonic acid amides; alkyl urea; the aryl urea; alkyl carbamate; aryl carbamate; heteroaryl; alkoxyl group; the alkoxyl group that replaces; halogenated alkoxy; alkylthio; the sulfo-haloalkyl; carboxyl; carbalkoxy; the alkyl carboxylic acid amides; the alkyl carboxylic acid amides that replaces; the dialkyl group carboxylic acid amides; the perhaps dialkyl group carboxylic acid amides of Qu Daiing.

14. compound, it is selected from compound 2,5 and 8 groups that constitute.

15. the compound of chemical formula (III) or its isomers, meta-bolites, many types of variant, prodrug or their salt.

Wherein:

R ₁Be hydrogen;

R ₂And R ₂₄Be independently of one another or jointly: alkyl, the alkyl that replaces, haloalkyl, thiazolinyl, the thiazolinyl that replaces, alkynyl, the alkynyl that replaces, acyl group, the acyl group that replaces, acyloxy, amino, single substituted-amino, disubstituted amido, alkyl sulfonamide, aryl sulfonic acid amides, alkyl urea, the aryl urea, alkyl carbamate, aryl carbamate, heteroaryl, alkoxyl group, the alkoxyl group that replaces, halogenated alkoxy, alkylthio, the sulfo-haloalkyl, carboxyl, carbalkoxy, the alkyl carboxylic acid amides, the alkyl carboxylic acid amides that replaces, the dialkyl group carboxylic acid amides, the perhaps dialkyl group carboxylic acid amides of Qu Daiing; And

R ₁₁And R ₁₂Be alkyl independently of one another or jointly, be selected from the group that constitutes by following group: methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, sec-butyl, the tertiary butyl, amyl group, tert-pentyl and n-pentyl.

16. medicinal compositions comprises in the claim 1 to 15 any one compound and pharmaceutically acceptable carrier or vehicle.

17. be used to regulate or the method for the kinase whose catalytic activity of arrestin, comprise that any one compound or the salt that makes in described protein kinase and the claim 1 to 15 contacts.

18. the method for claim 17, wherein said protein kinase are selected from the group that following material constitutes: receptor tyrosine kinase, nonreceptor tyrosine kinase and serine-threonine kinase.

19. the method for claim 17, the obstacle that wherein said protein kinase is relevant are to be selected from the relevant obstacle of the group that following obstacle constitutes: EGFR, the obstacle that PDGFR is relevant, relevant obstacle and the relevant obstacle of FLT-3 of obstacle, RET that cKit is relevant.

20. the method for claim 17, the obstacle that wherein said protein kinase is relevant are to be selected from the group that following cancer constitutes: squamous cell carcinoma, astrocytoma, Kaposi sarcoma, glioblastoma, lung cancer, bladder cancer, incidence cancer, melanoma, ovarian cancer, prostate cancer, mammary cancer, thyroid carcinoma, kidney, small cell lung cancer, leukemia, neurospongioma, colorectal carcinoma, apparatus urogenitalis cancer and gastrointestinal cancer.

21. the method for claim 17, the obstacle that wherein said protein kinase is relevant are to be selected from the group that following obstacle constitutes: diabetes, autoimmune disorder, hyper-proliferative obstacle, restenosis, fibrosis, psoriatic, von Heppel-Lindau disease, osteoarthritis, rheumatoid arthritis, blood vessel generation, inflammatory disorder, dysimmunity and cardiovascular disorder.

22. the method for claim 17, wherein said adjusting or inhibition occur in Mammals, and optional is the people.

23. the method for claim 18, wherein said kinases are in the intravital FLT-3 mutant of the patient who suffers from AML and/or cKit (KIT), or suffer from the mutant of the intravital cKit of patient of gastric and intestinal cancer or other cancers.

24. the method for claim 23, described adjusting or inhibition occur in suffer from cancer, optional be AML or gastric and intestinal cancer, mammalian body in, choose wantonly in people's body.