CN1784229A

CN1784229A - Heterocyclic kinase inhibitors: methods of use and synthesis

Info

Publication number: CN1784229A
Application number: CN 200480012042
Authority: CN
Inventors: M·A·西迪奎; D·贝兰格尔; C·戴; L·赵
Original assignee: Schering Corp
Current assignee: Merck Sharp and Dohme Corp
Priority date: 2003-03-10
Filing date: 2004-03-10
Publication date: 2006-06-07

Abstract

Inhibitors of kinases, compositions including the inhibitors, and methods of using the inhibitors and inhibitor compositions are described. The inhibitor s and compositions including them are useful for treating disease or disease symptoms. The invention also provides for methods of making kinase inhibitor compounds, methods of inhibiting kinase activity, and methods for treating disease or disease symptom.

Description

Heterocyclic kinase inhibitors: using method is with synthetic

The cross reference of related application

The application requires the U.S. Provisional Application series number 60/453 of application on March 10th, 2003,457, the U.S. Provisional Application series number 60/460 of application on April 7th, 2003,910, the U.S. Provisional Application series number 60/463 of application on April 15th, 2003, the U.S. Provisional Application series number 60/502 of 025 and 2003 on JIUYUE application in 12,, 710 priority above-mentionedly applies for that respectively integral body is attached to herein by reference.

Background of invention

Kinases is the enzyme with the transferance of catalysis phosphoryl.Protein kinase participates in the signal transduction activity of cell activation, growth and differentiation that control responds to the outer amboceptor of born of the same parents and environmental change.In general, kinases can be divided into several classes; The kinases of preferential phosphorylation serine and/or threonine residues is commonly referred to serine/threonine kinase, and the kinases of preferential phosphorylated tyrosine residue is commonly referred to tyrosine kinase [S.K.Hanks and T.Hunter, FASEB J., 1995,9,576-596].Tyrosine kinase comprises transmembrane growth factor receptor such as EGFR (S.Iwashita and M.Kobayashi, Cellular Signaling, 1992,4,123-132) and the non-receptor kinase of kytoplasm such as Lck, ZAP-70 and Syk kinases (C.Chan etc., Ann.Rev.Immun., 1994,12,555-592).

Many owing to the disorderly disease that causes of cell function all relates to unsuitable high protein kinase activity.Its direct or indirect occurrence cause for example may be because kinase whose correct control mechanism lost efficacy (for example relevant with sudden change, overexpression or the inappropriate activation of enzyme); Or because same cytokine or excessive generation of somatomedin or the generation deficiency that participates in the signal transduction in kinases upstream or downstream.In all these situations, kinase whose selectivity is suppressed to produce useful effect.

The T cell plays important effect in the initiation of transplant rejection, autoimmune disease and inflammatory reaction, be the main target that these symptom is carried out pharmaceutical intervention therefore.The activation of T cell is the complex process that causes cell growth and differentiation.The joint of TXi Baoshouti on sophisticated periphery T cell causes multiple intracellular signal, and the latter causes cell proliferation and obtains complicated function.Studied with the link coupled Biochemical Mechanism of incident in receptors bind and these born of the same parents (J.E.M., VanLeeuwen and L.E.Samelson, Current Opin.Immun.1999,11,242-248).The Syk family of tyrosine kinase (comprising Syk and ZAP-70) certain effect of performance in the initiation of receptor signal transduction with in amplifying (D.H.Chu etc., Immunol.Rev.1998,165,167-180).ZAP-70 only expresses in T cell and NK cell.Syk all has discovery in B cell, mastocyte, neutrophil cell, macrophage and platelet, it participates in B-cell receptor and the transduction of Fc receptor signal.Therefore, ZAP-70 and Syk inhibitors of kinases to treatment owing to the disease that T cell, NK cell, B cell, mastocyte, neutrophil cell, macrophage and hematoblastic activation and differentiation cause has potential curative effect.

Summary of the invention

The present invention relates to new compound and comprise these compound compositions, and the method for using and prepare chemical compound.Described chemical compound is a heterocyclic compound, can be used for therapeutic use, comprises disease or the disease symptoms regulated among the patient (for example mammal, people, Canis familiaris L., cat, horse).Described chemical compound can prepare separately, or prepares to obtain diversified library of compounds on structure and spatial chemistry in the mode of combination.Described chemical compound can be used as the inhibitor of these receptors by itself and the combining of ZAP-70 and Syk.

On the one hand, the present invention relates to formula (I) chemical compound

Formula (I)

Wherein,

A forms phenyl ring, pyridine ring, pyrimidine ring, thiphene ring, pyrrole ring, imidazole ring, pyrazoles ring, thiazole ring Huo oxazole ring;

X is O, S, NR ³, N (R ³) N (R ³), C (O), N (R ⁵) C (O), C (O) NR ⁵Or alkyl;

R ^XBe H, cycloalkyl, alkyl, heterocyclic radical, aryl, heteroaryl, aralkyl, heteroarylalkyl, alkynyl, aryl alkenyl, heteroaryl thiazolinyl, aromatic yl polysulfide yl, heteroaryl alkynyl, R ⁵O-alkyl, (R ⁵) ₃Si, acyl group, wherein R ^XOptional by 1-4 R ⁴Replace;

Each R ²Independently be H, NR ⁵ ₂, alkyl, cycloalkyl, thiazolinyl, alkynyl, heteroaryl, heterocyclic radical, aryl, halogen, acetyl group, aralkyl, heteroarylalkyl, aryl alkenyl, heteroaryl thiazolinyl, aromatic yl polysulfide yl, heteroaryl alkynyl, hydroxyl, alkoxyl, aryloxy group or heteroaryloxy, they are optional separately by 1-4 R ⁷Replace; Wherein work as X-R ^XDuring NH aryl that to be Me and Q replaced by heterocyclic radical, R ²Not Me; And wherein work as X-R ^XWhen being aryl alkenyl, R ²It or not acetyl group;

N is 0-3;

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁴Independently be halogen, OH, CF ₃, C (O) R ⁵, NR ³ ₂, N (R ³) C (O) R ⁵, CN, OCF ₃, SO ₂R ⁵Or SiR ⁵ ₃Perhaps alkoxyl, aryloxy group, alkyl, heterocyclic radical, R ⁵O-alkyl, cycloalkyl, aryl, alkylthio group, halogenated alkoxy, heteroaryl or aralkyl, they are optional separately by 1-4 R ⁶Replace;

Each R ⁵Independently be H or alkyl;

Each R ⁶Independently be halogen, OH, CF ₃, alkyl, alkoxyl, N (R ⁵)-alkyl, heteroaryl, heteroarylalkyl or heterocyclic radical;

Each R ⁷Independently be halogen, CN, OR ⁵, CF ₃, N (R ⁵) C (O) R ⁵, C (O) R ⁵, OCF ₃, SCF ₃, NR ⁵ ₂, C (O) NR ⁵ ₂, OH, R ⁵O-alkyl, alkyl, alkyl sulphonyl, heterocyclic radical or heteroaryl, they are optional separately by 1-4 R ⁸Replace;

Each R ⁸Independently be OR ⁵Or alkyl;

Q is H, halogen, C (O) R ⁵, C (O) R ⁹, C (S) R ⁵, C (S) R ⁹, C (O) NR ⁵ ₂, C (O) NR ⁵R ⁹, S (O) R ⁵, S (O) R ⁹, S (O) NR ⁵ ₂, S (O) NR ⁵R ⁹, SO ₂R ⁵, SO ₂R ⁹, SO ₂NR ⁵ ₂, SO ₂NR ⁵R ⁹, NR ⁵ ₂, NR ⁵R ⁹, R ⁹S-alkyl, alkyl or heterocyclic radical, wherein each alkyl or heterocyclic radical are optional by 1-4 R ¹⁰Replace;

Each R ⁹Independently be aryl, heterocyclic radical, heteroaryl, aralkyl or heteroarylalkyl, they are optional separately by 1-4 R ¹⁰Replace;

Each R ¹⁰Independently be alkyl, CF ₃, C (NH) NR ⁵R ¹¹, C (NH) R ¹¹, CN, R ⁵ ₂N-alkyl, NR ⁵R ¹¹-alkyl, R ⁵O-alkyl, R ¹¹, heteroaryl, heterocyclic radical or heterocyclic radical alkyl, they are optional separately by alkyl or OR ⁵Replace;

Each R ¹¹Independently be aralkyl, heteroarylalkyl, cycloalkyl or heterocyclic radical.

In some cases, A forms phenyl ring.

On the other hand, the present invention relates to formula (II) chemical compound

Formula (II)

Wherein,

Each R ²Independently be H, NR ⁵ ₂, alkyl, cycloalkyl, thiazolinyl, alkynyl, heteroaryl, heterocyclic radical, aryl, halogen, acetyl group, aralkyl, heteroarylalkyl, aryl alkenyl, heteroaryl thiazolinyl, aromatic yl polysulfide yl, heteroaryl alkynyl, hydroxyl, alkoxyl, aryloxy group or heteroaryloxy, they are optional separately by 1-4 R ⁷Replace;

Each R ^{2 '}Independently be H, halogen, NH ₂, alkyl, OH, C (O) Me, aryl, heteroaryl;

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁸Independently be OR ⁵Or alkyl;

Het is optional by 1-4 R ¹⁰The heterocyclic radical that replaces;

In some cases, Het connects by theheterocyclic nitrogen atom.

In some cases, R ^{2 '}Be H.

In some cases, Het connects by theheterocyclic nitrogen atom; X is NR ³Or alkyl; R ^XBe cycloalkyl, aryl, heteroaryl, aralkyl, heteroarylalkyl or heterocyclic radical, wherein R ^XOptional by 1-4 R ⁴Replace; R ^{2 '}Be H.

In some cases, Het is

Or

In other cases, Het is

On the other hand, the present invention relates to formula (III) chemical compound

Formula (III)

Wherein,

Each R ²Independently be H, NR ⁵ ₂, alkyl, cycloalkyl, thiazolinyl, alkynyl, heteroaryl, heterocyclic radical, aryl, halogen, acetyl group, aralkyl, heteroarylalkyl, aryl alkenyl, heteroaryl thiazolinyl, aromatic yl polysulfide yl, heteroaryl alkynyl, hydroxyl, alkoxyl, aryloxy group or heteroaryloxy, they are optional separately by 1-4 R ⁷Replace; Wherein work as X-R ^XBe Me and R ¹²During the aryl that replaced by heterocyclic radical, R ²Not Me;

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁸Independently be OR ⁵Or alkyl;

Each R ¹²Independently be optional by 1-4 R ¹³The aryl or the heteroaryl that replace;

Each R ¹³Independently be optional by alkyl or OR ⁵The heterocyclic radical that replaces.

In some cases, R ^{2 '}Be H.

In some cases, R ¹²Be

Or

On the other hand, the present invention relates to formula (IV) chemical compound

Formula (IV)

Wherein,

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁸Independently be OR ⁵Or alkyl;

Each R ¹¹Independently be aralkyl, heteroarylalkyl, cycloalkyl or heterocyclic radical;

R ¹⁴Be halogen, NH ₂, alkyl, OH, C (O) Me, aryl, heteroaryl or C (O) NHR ⁵

Another aspect the present invention relates to the formula V chemical compound

Formula V

Wherein,

R ^YBe H, halogen, aryl, heteroaryl, cycloalkyl, heterocyclic radical, alkynyl, aryl alkenyl, heteroaryl thiazolinyl, aromatic yl polysulfide yl or heteroaryl alkynyl, wherein R ^YOptional by 1-4 R ⁴Replace;

Each R ²Independently be H, NR ⁵ ₂, alkyl, cycloalkyl, thiazolinyl, alkynyl, heteroaryl, heterocyclic radical, aryl, halogen, acetyl group, aralkyl, heteroarylalkyl, aryl alkenyl, heteroaryl thiazolinyl, aromatic yl polysulfide yl, heteroaryl alkynyl, hydroxyl, alkoxyl, aryloxy group or heteroaryloxy, they are optional separately by 1-4 R ⁷Replace; Wherein work as R ^YWhen the indole that is halogen, is replaced by halogen, phenyl or the phenyl that replaced by halogen, R ²Not Me; Wherein work as R ^YDuring the phenyl that replaced by halogen, R ²It or not isopropyl; Wherein work as R ^YDuring the indole that replaced by halogen, R ²The alkynyl that is replaced by heterocyclic radical not; Wherein work as R ^YWhen being H, R ²Not phenyl, wherein work as R ^YWhen indole that is replaced by halogen or the phenyl that replaced by halogen, R ²It or not halogen; And wherein work as R ^YWhen being phenyl or substituted-phenyl, R ²It or not acetyl group;

N is 0-3;

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁸Independently be OR ⁵Or alkyl;

In some cases, A forms phenyl ring.

On the one hand, the present invention relates to formula (VI) chemical compound

Formula (VI)

Wherein,

Each R ²Independently be H, NR ⁵ ₂, alkyl, cycloalkyl, thiazolinyl, alkynyl, heteroaryl, heterocyclic radical, aryl, halogen, acetyl group, aralkyl, heteroarylalkyl, aryl alkenyl, heteroaryl thiazolinyl, aromatic yl polysulfide yl, heteroaryl alkynyl, hydroxyl, alkoxyl, aryloxy group or heteroaryloxy, they are optional separately by 1-4 R ⁷Replace; Wherein work as R ^YWhen the indole that is halogen, is replaced by halogen, phenyl or the phenyl that replaced by halogen, R ²Not Me; Wherein work as R ^YDuring the phenyl that replaced by halogen, R ²It or not isopropyl;

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁸Independently be OR ⁵Or alkyl;

Het is optional by 1-4 R ¹⁰The heterocyclic radical that replaces;

In some cases, Het connects by theheterocyclic nitrogen atom; R ²Not H; R ^{2 '}Be H.

In some cases, Het is

Or

In some cases, Het is

On the other hand, the present invention relates to formula (VII) chemical compound

Formula (VII)

Wherein,

Each R ²Independently be H, NR ⁵ ₂, alkyl, cycloalkyl, thiazolinyl, alkynyl, heteroaryl, heterocyclic radical, aryl, halogen, acetyl group, aralkyl, heteroarylalkyl, aryl alkenyl, heteroaryl thiazolinyl, aromatic yl polysulfide yl, heteroaryl alkynyl, hydroxyl, alkoxyl, aryloxy group or heteroaryloxy, they are optional separately by 1-4 R ⁷Replace; Wherein work as R ^YWhen being H, R ²It or not phenyl;

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁸Independently be OR ⁵Or alkyl;

In some cases, R ²Not H; R ^{2 '}Be H.

In some cases, R ¹²Be

Or

On the one hand, the present invention relates to formula (VIII) chemical compound

Formula (VIII)

Wherein,

Each R ²Independently be H, NR ⁵ ₂, alkyl, cycloalkyl, thiazolinyl, alkynyl, heteroaryl, heterocyclic radical, aryl, halogen, acetyl group, aralkyl, heteroarylalkyl, aryl alkenyl, heteroaryl thiazolinyl, aromatic yl polysulfide yl, heteroaryl alkynyl, hydroxyl, alkoxyl, aryloxy group or heteroaryloxy, they are optional separately by 1-4 R ⁷Replace; Wherein work as R ^YWhen indole that is halogen, is replaced by halogen or the optional phenyl that is replaced by halogen, R ²Not Me; Wherein work as R ^YDuring the phenyl that replaced by halogen, R ²It or not isopropyl; Wherein work as R ^YDuring the indole that replaced by halogen, R ²The alkynyl that is replaced by heterocyclic radical not; Wherein work as R ^YWhen being H, R ²It or not phenyl; And wherein work as R ^YWhen being phenyl or substituted-phenyl, R ²It or not acetyl group;

Each n is 0-3;

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁸Independently be OR ⁵Or alkyl;

In some cases, n is 1, R ²Not H.

On the other hand, the present invention relates to formula (XIV) chemical compound

Formula (XIV)

Wherein,

Each R ²Independently be H, NR ⁵ ₂, alkyl, cycloalkyl, thiazolinyl, alkynyl, heteroaryl, heterocyclic radical, aryl, halogen, aralkyl, heteroarylalkyl, aryl alkenyl, heteroaryl thiazolinyl, aromatic yl polysulfide yl, heteroaryl alkynyl, aryloxy group or heteroaryloxy, they are optional separately by 1-4 R ⁷Replace;

N is 0-3;

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁸Independently be OR ⁵Or alkyl;

Another aspect the present invention relates to the method for preparation formula (II) chemical compound

Formula (II)

Wherein X, R ^X, R ², R ^{2 '}Be defined as follows with Het.Described method comprises:

Handle formula (IX) chemical compound with malonic acid, obtain formula (X) ring expansion chemical compound;

Formula (IX) formula (X)

Make formula (X) chemical compound and Pd catalyst and the coupling of formula (XI) chemical compound, obtain formula (XII) chemical compound;

Formula (X) formula (XI) formula (XII)

Use POCl ₃Processing formula (XII) chemical compound obtains formula (XIII) chloride, makes the amine coupling of formula (XII) carboxylic acid and formula Het-H simultaneously, obtains formula (XIII) chemical compound;

Formula (XII) formula (XIII)

Make formula (XIII) chemical compound and one or more coupling agent couplings, obtain formula (II) chemical compound,

Formula (XIII) formula (II)

Wherein for formula II and formula IX-XIII,

X is O, S, NR ³, N (R ³) N (R ³), C (O), N (R ⁵) C (O) R ⁵, C (O) NR ⁵Or alkyl;

Each R ²Independently be NR ⁵ ₂, alkyl, cycloalkyl, thiazolinyl, alkynyl, heteroaryl, heterocyclic radical, aryl, halogen, aralkyl, heteroarylalkyl, aryl alkenyl, heteroaryl thiazolinyl, aromatic yl polysulfide yl, heteroaryl alkynyl, aryloxy group or heteroaryloxy, they are optional separately by 1-4 R ⁷Replace;

R ^{2 '}Be H, halogen, NH ₂, alkyl, OH, C (O) Me, aryl, heteroaryl;

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁸Independently be OR ⁵Or alkyl;

Het is optional by 1-4 R ¹⁰The heterocyclic radical that replaces;

In some cases, the H of Het-H is connected with nitrogen.

In some cases, coupling agent is H ₂NR ^X

In some cases, coupling agent is MgCl-R ^XAnd Pd.

In some cases, coupling agent is HSR ^X

On the other hand, the present invention relates to treat the method for autoimmune disease among the patient, described method comprises and gives patient's any chemical compound as herein described or compositions.

In some cases, described method comprises and gives other medicine.

In some cases, autoimmune disease is a lupus.

On the other hand, the present invention relates to treat the method for organ-graft refection among the patient, described method comprises and gives patient's any chemical compound as herein described or compositions.

On the other hand, the present invention relates to treat the method for inflammatory diseases among the patient, described method comprises and gives patient's any chemical compound as herein described or compositions.

In some cases, described method comprises and gives other medicine.Described other medicine can be analgesic or steroid.

In some cases, inflammatory diseases is an arthritis.Arthritis can be for example rheumatoid arthritis, rheumatoid spondylitis, gouty arthritis, traumatic arthritis, rubella arthritis, arthritic psoriasis or osteoarthritis.

In some cases, inflammatory diseases is inflammatory bowel or Crohn disease.

On the other hand, the present invention relates to comprise any compound compositions as herein described.

In some cases, described compositions can comprise drug acceptable carrier.

In some cases, described compositions can comprise other medicine.

On the other hand, the present invention relates to the library of any formula (I)-(VIII) chemical compound.

Another aspect the present invention relates to suppress the method that IL-2 generates among the patient, and described method comprises and gives patient's any chemical compound as herein described or compositions.

Also on the one hand, the present invention relates to regulate the method for ZAP-70 among the patient or Syk, described method comprises and gives patient's any chemical compound as herein described or compositions.

In other respects, chemical compound described herein, compositions and method are any chemical compound of this paper table 1.

The detailed content of one or more embodiments of the invention is illustrated in the following drawings and explanation.According to explanation and accompanying drawing and claims, other characteristics of the present invention, target and advantage are conspicuous.

Detailed Description Of The Invention

Term " halogen " refers to the group of any fluorine, chlorine, bromine or iodine.Term " alkyl " refers to contain the straight or branched hydrocarbon chain of indicated number purpose carbon atom.For example, C ₁-C ₁₀Can contain 1-10 (comprising 10) carbon atom in the expression group.Term " low alkyl group " refers to C ₁-C ₈Alkyl chain.Do not occurring under any figure notation situation, " alkyl " is the chain (straight or branched) that wherein contains 1-10 (comprising 10) carbon atom.Term " alkoxyl " refers to-the O-alkyl group.Term " alkylidene " refers to that divalent alkyl (promptly-R-).Term " alkylidene dioxygen generation " refers to bivalence-O-R-O-structure, and wherein R represents alkylidene.Term " aminoalkyl " refers to the alkyl that replaced by amino.Term " sulfydryl " refers to-the SH group.Term " thio alkoxy " refers to-the S-alkyl group.

Term " thiazolinyl " refers to have the straight or branched hydrocarbon chain of one or more carbon-to-carbon double bonds.Alkenyl part contains indicated number purpose carbon atom.For example, C ₂-C ₁₀Can contain 2-10 (comprising 10) carbon atom in the expression group.Term " low-grade alkenyl " refers to C ₂-C ₈Alkenylene chain.Do not occurring under any figure notation situation, " thiazolinyl " is the chain (straight or branched) that wherein contains 2-10 (comprising 10) carbon atom.

Term " alkynyl " refers to have the straight or branched hydrocarbon chain of one or more carbon-to-carbon triple bonds.Alkynyl partly contains indicated number purpose carbon atom.For example, C ₂-C ₁₀Can contain 2-10 (comprising 10) carbon atom in the expression group.Term " low-grade alkynyl " refers to C ₂-C ₈The alkynyl chain.Do not occurring under any figure notation situation, " alkynyl " is the chain (straight or branched) that wherein contains 2-10 (comprising 10) carbon atom.

Term " aryl " refers to 6-carbon monocycle or 10-carbon dicyclo aromatic ring system, and wherein 0,1,2,3 or 4 of each ring atom can be substituted the base replacement.The example of aryl comprises phenyl, naphthyl etc.Term " aralkyl " refers to the alkyl that replaced by aryl.Term " aryl alkenyl " refers to the thiazolinyl that replaced by aryl.Term " aromatic yl polysulfide yl " refers to the alkynyl that replaced by aryl.Term " alkoxy aryl " refers to the alkoxyl that replaced by aryl.

Term used herein " cycloalkyl " comprise have 3-12 carbon, preferred 3-8 carbon, the more preferably saturated and part unsaturated cyclic alkyl of 3-6 carbon, wherein cycloalkyl can be chosen wantonly and be substituted.Preferred cycloalkyl includes but not limited to cyclopropyl, cyclobutyl, cyclopenta, cyclopentenyl, cyclohexyl, cyclohexenyl group, suberyl and ring octyl group.

Term " heteroaryl " refers to have 1-3 hetero atom (for single loop system), the aromatics 5-8 of a 1-6 hetero atom (for bicyclic system) or 1-9 hetero atom (for three-loop system) unit monocycle, 8-12 unit's dicyclo or 11-14 unit three ring ring systems, described hetero atom (for example is selected from O, N or S, for single loop system, bicyclic system or three-loop system, contain carbon atom and 1-3, a 1-6 or 1-9 N, O or S hetero atom respectively), wherein 0,1,2,3 or 4 of each ring atom can be substituted the base replacement.The example of heteroaryl comprises pyridine radicals, furyl, imidazole radicals, benzimidazolyl, pyrimidine radicals, thienyl, quinolyl, indyl, thiazolyl etc.Term " heteroarylalkyl " refers to the alkyl that replaced by heteroaryl.Term " heteroaryl thiazolinyl " refers to the thiazolinyl that replaced by heteroaryl.Term " heteroaryl alkynyl " refers to the alkynyl that replaced by heteroaryl.Term " heteroaryl alkoxyl " refers to the alkoxyl that replaced by heteroaryl.

Term " heterocyclic radical " refers to have 1-3 hetero atom (for single loop system), non-aromatics 5-8 unit monocycle, 8-12 unit's dicyclo or the 11-14 unit three ring ring systems of a 1-6 hetero atom (for bicyclic system) or 1-9 hetero atom (for three-loop system), described hetero atom (for example is selected from O, N or S, for single loop system, bicyclic system or three-loop system, contain carbon atom and 1-3, a 1-6 or 1-9 N, O or S hetero atom respectively), wherein 0,1,2 or 3 of each ring atom can be substituted the base replacement.The example of heterocyclic radical comprises piperazinyl, pyrrolidinyl, alkyl dioxin, morpholinyl, tetrahydrofuran base etc.Term " heterocyclic radical alkyl " refers to the alkyl that replaced by heterocyclic radical.

Term " oxo " refers to oxygen atom, forms carbonyl when it is connected with carbon, forms the N-oxide when being connected with nitrogen, forms sulfoxide or sulfone when being connected with sulfur.

Term " acyl group " refers to alkyl-carbonyl, naphthene base carbonyl, aryl carbonyl, heterocyclic radical carbonyl or heteroaryl carbonyl substituted base, and their any bases that all can further be substituted replace.

Term " sulfonyl " refers to the sulfur that is connected with two oxygen atoms by two keys." alkyl sulphonyl " refers to the alkyl that replaced by sulfonyl.

Term " substituent group " refers to the group that carries out " replacement " on any atom of alkyl, cycloalkyl, aryl, heterocyclic radical or heteroaryl.Suitable substituents includes but not limited to halogen, hydroxyl, sulfydryl, oxo, nitro, haloalkyl, alkyl, aryl, aralkyl, alkoxyl, thio alkoxy, aryloxy group, amino, alkoxy carbonyl, amide groups, carboxyl, alkyl sulphonyl, alkyl-carbonyl and cyano group.

Term " Me " fingernail base.

Term " Ac " refers to acetyl group.

Term " dppf " refers to 1, two (diphenylphosphine) ferrocene of 1-.

Term " Mops " refers to 3-(N-morpholino) propane sulfonic acid.

Term " BSA " refers to bovine serum albumin.

Term " LAT " refers to the palmitoylation p36/38kDa plasma membrane associated protein expressed in all T lymphocytes, it is the physiological substrate of activation ZAP-70.

Term " TMB " refers to 3,3 ', 5,5 '-tetramethyl benzidine.

The disease of signal conduction, mediation, adjusting or the control of term " kinase mediated disease " finger protein kinases involved in diseases process or symptom.The example of kinase mediated disease is by following a few class disease examples: cancer, autoimmune disease, metabolic disease, inflammatory diseases, infectious disease (infection such as antibacterial, virus, yeast, fungus), central nervous system disease, degenerative neurological disorder, allergy/asthma, dermatosis, angiogenesis disease, neovascularization disease, blood vessel generation disease, cardiovascular disease etc.

Table 1: representative compounds of the present invention ^*

The sequence number molecular structure

^*In the above structure, do not mark the hydrogen atom that is connected with secondary nitrogen-atoms.

Chemical compound as herein described, compositions and method can be used for suppressing ZAP-70 and Syk.Therefore, described chemical compound, compositions and method can be used for treating ZAP-70 and different disease or the disease symptoms of Syk Jie in the mammal, the particularly mankind.ZAP-70 and Syk are situated between different disease by following example: lupus, organ-graft refection and inflammatory diseases.The example of inflammatory diseases comprises arthritis (for example rheumatoid arthritis, rheumatoid spondylitis, gouty arthritis, traumatic arthritis, rubella arthritis, arthritic psoriasis or osteoarthritis), inflammatory bowel and Crohn disease.

Chemical compound of the present invention, compositions and method can be used for treating for example lupus, organ-graft refection's (for example kidney, liver, heart, lungs, pancreas (islet cells), bone marrow, cornea, small intestinal, skin alloplast or xenograft) and inflammatory diseases.The example of inflammatory diseases includes but not limited to arthritis (for example rheumatoid arthritis, rheumatoid spondylitis, gouty arthritis, traumatic arthritis, rubella arthritis, arthritic psoriasis or osteoarthritis), asthma, allergy, eczema, inflammatory bowel and Crohn disease.

Others of the present invention relate to the compositions that comprises any chemical compound described herein and drug acceptable carrier; Or comprise any chemical compound described herein, other medicine (for example anti-inflammatory drug, nonsteroidal anti-inflammatory drug (NSAID), steroid etc.) and the compositions of drug acceptable carrier; Or this paper comprises any chemical compound described herein, other medicine and the compositions of drug acceptable carrier, and wherein other medicine is kinases bonding agent (for example polypeptide, antibody or an organic molecule).

Another aspect of the invention relates to the method for the treatment of the patient (for example mammal) who suffers from kinase mediated disease or disease symptoms (including but not limited to pain and inflammation).Described method comprises and gives patient's (comprising the patient who is accredited as this treatment of needs) chemical compound described herein of effective dose, maybe can produce the compositions described herein of this effect.Can judge identify needing the patient of this treatment, institute does to judge can be subjective (for example subjective suggestion) or objectively (for example can by testing or diagnostic method mensuration) by patient or health care professional.

The invention further relates to the product of making by said method (being the chemical compound of any formula of this paper).

The substituent group of the present invention's prospect and the combination of variable only are the combinations that causes forming stable compound.Term used herein " stable " refers to that the stability that this paper chemical compound has is enough to allow to produce, and can keep the sufficiently long time of chemical compound integrity to be used for the purpose (for example the patient being carried out therapeutic or preventive administration) that this paper describes in detail.

Synthesizing of heterocyclic kinase inhibitors

Formula (II) (variable of wherein various (for example formula (II)) as defined herein) chemical compound is prepared as follows: handle formula (IX) chemical compound with malonic acid, obtain the ring expansion chemical compound, the latter follows and contains R ²The chemical compound coupling, obtain formula (XII).

Formula (IX) formula (XII)

Formula (XII) chemical compound chlorinating agent such as POCl ₃Handle, gained carboxylic acid and heterocycle (for example heteroaryl or heterocyclic radical) chemical compound (" Het ") coupling obtains formula (XIII) chemical compound.

Formula (XII) formula (XIII)

Gained chemical compound and one or more coupling agent couplings then obtains formula (II) chemical compound.

Formula (XIII) formula (II)

Though the foregoing description describes with quinoline nuclei, also can use other heterocyclic nucleus.For example, above-mentioned synthetic can be by carrying out with the above-mentioned formula of following exemplary heterocyclic substituted (IX) raw material:

These chemical compounds only are used for describing, and do not have a mind to limit the scope of synthetic schemes.

Term " coupling agent " refers to the chemical reagent that uses in reaction, it makes between certain chemical part and another part from coupling agent and forms key.Some example of coupling agent comprises transition metal such as Pd, Cu and Mg and transition-metal catalyst, also comprises boron-containing compound such as borine.Coupling agent also comprises nucleopilic reagent such as amine, alkoxide, sulfide or corresponding protonated form.

Those skilled in the art can recognize that the more multi-method of the synthetic various chemical compound of this paper is conspicuous to those of ordinary skills.In addition, different synthesis steps can other order or the order carry out, to obtain required chemical compound.The synthetic chemistry conversion method and the protecting group methodology (protection and deprotection) that are used for synthetic chemical compound described herein are well known in the art, comprise for example at R.Larock, Comprehensive Organic Transformations, VCHPublishers (1989); T.W.Greene and P.G.M.Wuts, Protective Groups inOrganic Synthesis, the 2nd edition, John Wiley and Sons (1991); L.Fieser and M.Fieser, Fieser and Fieser ' s Reagents for Organic Synthests, John Wileyand Sons (1994); And L.Paquette (editor), Encyclopedia of Reagents forOrganic Synthesis, the method for describing in John Wiley and Sons (1995) and follow-up each edition thereof.

The compounds of this invention can contain one or more asymmetric centers, therefore can racemate and racemic mixture, simple enantiomer, independent diastereomer and the form of non-enantiomer mixture occur.All these isomeric forms of these chemical compounds obviously comprise in the present invention.The compounds of this invention also can show as multiple tautomeric forms, in this case, the present invention obviously comprises all tautomeric forms (for example the alkylation of loop systems causes in a plurality of positions alkylation taking place, and the present invention obviously comprises all this product) of chemical compound described herein.This class isomeric form of all of this chemical compound obviously comprises in the present invention.The all crystals form of chemical compound described herein obviously comprises in the present invention.

The compounds of this invention used herein comprises various chemical compound described herein, is defined as to comprise that its medicine can accept derivant or prodrug." medicine can be accepted derivant or prodrug " refers to any drug acceptable salt, ester, ester salt or other derivant of this paper chemical compound, and it can (directly or indirectly) provide The compounds of this invention when giving the receiver.Particularly preferred derivant and prodrug are the derivant and the prodrugs of the bioavailability (for example more easily being absorbed into blood by the chemical compound that makes orally give) that can increase The compounds of this invention when giving mammal with these chemical compounds, or can promote derivant and the prodrug that parent compound is sent to biological compartment (for example brain or lymphsystem) with respect to parent compound.Preferred prodrug comprises can strengthen water solublity or the group by the goldbeater's skin Active transport appends to various structural derivant described herein.

The compounds of this invention can be modified by additional appropriate functional degree, to strengthen the selectivity organism characteristic.These modifications are well known in the art, comprise enhancing to specific biological compartment (for example blood, lymphsystem, central nervous system's) biological penetrance, increase oral availability, increase dissolubility and make and can be used for drug administration by injection, change metabolism and change the modification of discharge rate.

The drug acceptable salt of The compounds of this invention comprises the salt that is derived from the acceptable inorganic and organic acids and base of medicine.The example of suitable hydrochlorate comprises acetate, adipate, benzoate, benzene sulfonate, butyrate, citrate, digluconate, dodecyl sulphate, formates, fumarate, glycollate, Hemisulphate, enanthate, caproate, hydrochlorate, hydrobromate, hydriodate, lactate, maleate, malonate, mesylate, the 2-naphthalene sulfonate, nicotinate, nitrate, palmoate, phosphate, picrate, pivalate, propionate, Salicylate, succinate, sulfate, tartrate, toluene fulfonate and hendecane hydrochlorate.The salt that is derived from appropriate base comprises alkali metal (for example sodium) salt, alkaline-earth metal (for example magnesium) salt, ammonium salt and N-(alkyl) ₄ ⁺Salt.The present invention also looks forward to come into the open any alkaline nitrogen-containing group quaternized of compound to this paper.Water solublity or oil-soluble or dispersibility product can be by this quaternized acquisitions.

Various chemical compound described herein can for example pass through under intravenous, intra-arterial, the corium, intraperitoneal, intramuscular or subcutaneous injection administration; Or oral, suck, per nasal, through mucous membrane, as ophthalmic preparation topical or inhalation, dosage is about 0.001 to about 100mg/ kg body weight, preferred dose is every 4-120 hour 10mg-1000mg/ clothes, or decides according to the requirement of concrete medicine.The expection of the method for this paper gives the chemical compound of effective dose or compound composition to obtain effect required or regulation.Usually, about 1 time to about 6 times of pharmaceutical composition of the present invention administration every day is perhaps as infusion solution administration continuously.This administration can be used for chronic or acute treatment.The amount with the active component that produces the single dose form of can combining with carrier material depends on the patient that receives treatment and concrete administering mode and changes.Representational preparation contains about 5% to about 95% reactive compound (weight).Perhaps, this preparation contains about 20% to about 80% reactive compound.

Might need the dosage lower or higher than above-mentioned dosage.The concrete dosage and the therapeutic scheme that are used for any concrete patient depend on multiple factor, comprise that the order of severity of activity, age, body weight, general health situation, sex, diet, administration time, excretion rate, composite reagent situation, disease or symptom of used particular compound and process, patient are to the disposal of disease or symptom and treatment doctor's judgement suggestion.

Status of patient can give chemical compound of the present invention, compositions or their combination of maintenance dose after improving in case of necessity.Subsequently, decide on the symptom situation, both can be reduced to the level of keeping the improvement state dosage or frequency or they.But, one has the recurrence of disease symptoms, and needs of patients carries out intermittent therapy for a long time.

Pharmaceutical composition of the present invention comprises various chemical compound described herein or the acceptable salt of its medicine; Other medicine comprises for example steroid or analgesic; And any drug acceptable carrier, adjuvant or excipient.Perhaps the present composition comprises various chemical compound described herein or the acceptable salt of its medicine; Medicine acceptable carrier, adjuvant or excipient.Compositions as herein described comprises various chemical compound described herein, and other medicine, described medicine if present, its quantity can effectively realize the adjusting to disease or disease symptoms, comprises the adjusting of kinase mediated disease or its symptom.The preparation method for compositions comprises one or more chemical compounds as herein described and one or more carriers, and optional and the bonded step of one or more other medicines as herein described.

Term " drug acceptable carrier or adjuvant " refers to and can give the patient with The compounds of this invention, does not influence the pharmacological activity of described chemical compound, and when being enough to transmit the dosage of therapeutic dose chemical compound nontoxic carrier or adjuvant.

Pharmaceutical composition can be the form of sterile injectable agent, and for example sterile injectable contains aqueous suspension agent or contains oil-suspending agent.This suspending agent can use the preparation of suitable dispersant or wetting agent (for example Tween 80) and suspending agent by technology well known in the art.The sterile injectable agent also can be that nontoxic parenteral can be accepted sterile injectable solution agent or the suspending agent in diluent or the solvent, for example solution of 1,3 butylene glycol.Operablely accept excipient or solvent has mannitol, water, Ringer's solution and isotonic sodium chlorrde solution.In addition, aseptic, not volatile oil preparation is usually as solvent or suspension media.For this reason, any nonirritant, not volatile oil be can use, synthetic monoglyceride or diglyceride comprised.Fatty acid for example oleic acid and glyceride ester derivatives thereof can be used for the preparation of injectable agent, also can use natural medicine can accept oils for example olive oil or Oleum Ricini, particularly their polyoxyethylene form.These oily solution or suspending agent also can contain long-chain alcohols diluent or dispersant, or carboxymethyl cellulose or be generally used for the compounding pharmaceutical acceptable forms such as the similar dispersant of emulsion agent and/or suspending agent.Other conventional surfactant for example tween or department class and/or be usually used in prepares medicine and can accept other similar emulsifying agent of solid, liquid or other dosage form or bioavailability reinforcing agent and also can be used for the purpose prepared.

Pharmaceutical composition of the present invention can any oral acceptable forms oral administration, includes but not limited to capsule, tablet, Emulsion and aqueous suspension agent, dispersant and solution.If oral tablet, carrier commonly used comprises lactose and corn starch.Usually also add lubricant such as magnesium stearate.For oral capsule, useful diluent comprises lactose and dried corn starch.When aqueous suspension agent and/or Emulsion were used for oral administration, active component can be suspended in or is dissolved in the oil phase with emulsifying agent or suspending agent.If desired, can add some sweeting agent and/or correctives and/or coloring agent.

Pharmaceutical composition of the present invention also can the suppository form rectally.These compositionss can prepare by The compounds of this invention is mixed with suitable nonirritant excipient, and described excipient is solid when room temperature, is liquid when rectal temperature, therefore can melt release of active ingredients at rectum.These excipient materials include but not limited to cupu oil, Cera Flava, Polyethylene Glycol.

The drug acceptable carrier that can be used for pharmaceutical composition of the present invention, adjuvant and excipient include but not limited to ion-exchanger, aluminium oxide, aluminium stearate, lecithin, self-emulsifying drug delivery system (SEDDS) is d-alpha-tocopherol cetomacrogol 1000 succinate for example, the surfactant such as tween or other the similar polymer release matrix that are used for pharmaceutical dosage form, serum albumin such as human serum albumin, buffer substance such as phosphate, glycine, sorbic acid, potassium sorbate, the partial glycerol ester admixture of saturated vegetable fatty acid, water, salt or electrolyte such as protamine sulfate, sodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, silica gel, magnesium trisilicate, polyvinylpyrrolidone, based on cellulosic material, Polyethylene Glycol, sodium carboxymethyl cellulose, polyacrylate, wax, polyethylene-polyoxypropylene block polymer, Polyethylene Glycol and lanoline.Cyclodextrin such as α-, β-and gamma-cyclodextrin also can advantageously be used for strengthening the release of various chemical compound described herein.

In some cases, the pH of prescription can use the acceptable acid of medicine, alkali or buffer substance adjustment, to improve the stability of compound formula or its releasing pattern.

That term used herein " parenteral " comprises is subcutaneous, in the Intradermal, intravenous, intramuscular, intraarticular, intra-arterial, synovial fluid, in the breastbone, in the sheath, intralesional, intracranial injection or infusion techn.

When required treatment related to topical easy to reach position or organ, pharmaceutical composition of the present invention can be applied by topical.When the part was used for skin, pharmaceutical composition should wherein contain the active component that is suspended in or is dissolved in carrier with suitable ointment preparation.The carrier that is used for the The compounds of this invention topical includes but not limited to mineral oil, liquid paraffin, white vaseline, propylene glycol, polyoxyethylene polyoxypropylene complex, emulsifing wax and water.In addition, pharmaceutical composition can wherein contain the reactive compound that is suspended in or is dissolved in carrier with suitable emulsifying agent with suitable lotion or Emulsion preparation.Suitable carriers includes but not limited to mineral oil, sorbitan monostearate, polysorbate60, cetyl esters wax, cetearyl alcohol, 2-octyl dodecanol, benzyl alcohol and water.Pharmaceutical composition of the present invention also can rectal suppository or suitable enema forms at the lower intestinal tract topical.Local percutaneous patch is also included within the present invention.

Pharmaceutical composition of the present invention can pass through nose aerosol or inhalant administration.This compositions can prepare with the technology that field of pharmaceutical preparations is known, can adopt absorption enhancer, fluorocarbon and/or other solubilizing agents well known in the art or the dispersant of benzylalcohol or other suitable antiseptic, raising bioavailability, be prepared into the saline solution agent.

When the present composition comprises the combination of various chemical compound described herein and one or more other therapeutic or preventive medicine, the dosage level of chemical compound and other medicine should be about 1-100% of single therapy scheme bio-occlusion pharmaceutical quantities, more preferably from about 5-95%.Other medicine can be used as the part of multiple dosage, separates separately giving with The compounds of this invention.Perhaps, these medicines can be used as the part of single dosage form, are mixed in the single compositions with The compounds of this invention.

Embodiment:

Synthesizing of embodiment 1:2-quinolin-2-ylamine

Make 5-iodine isatin (10g, 36.3mmol) and malonic acid (7.5g, 72mmol) 200mL glacial acetic acid solution refluxes and spends the night.Filter collecting precipitation, with AcOH and washing with acetone.Then with EtOH backflow gained solid 1h.Filter, with EtOH and Et ₂The O washing obtains product 6-iodo-2-oxo-1,2-dihydro-quinoline-4-formic acid 2, yield 8.8g (76%). ¹H-NMR(400MHz，DMSO-d ₆)：δ14.0(br?s，1H)，12.13(s，1H)，8.56(d，1H，J＝8.1Hz)，7.83(dd，1H，J＝8.7，1.8Hz)，7.17(d，1H，J＝8.4Hz)，6.93(s，1H)。

Make 6-iodo-2-oxo-1, and 2-dihydro-quinoline-4-formic acid 2 (3.15g, 10mmol), 3,4-(methylene-dioxy) phenylboric acid (2.49g, 15mmol), K ₃PO ₄(8.49g, 40mmol) and Pd (OAc) ₂(112mg is 0.5mmol) at 60mL degassing H ₂Mixture among the O heats 2h under 60 ℃ and argon atmospher.After being chilled to room temperature, solid collected by filtration is used H ₂O and washing with acetone.Handle with 20mL 1M HCl then, refilter gained green-yellow solid, use H ₂The O washing.Use P ₂O ₅Vacuum drying obtains 2.53g (82%) product 3, is the green-yellow solid. ¹H-NMR(400MHz，DMSO-d ₆)：δ14.0(brs，1H)，12.11(s，1H)，834(s，1H)，7.79(d，IH，J＝7.6Hz)，7.40(d，1H，J＝8.6Hz)，7.18(s，1H)，7.08(d，1H7?J＝7.1Hz)，7.01(d，1H，J＝8.2Hz)，6.91(s，1H)，6.06(s，2H)。

Make 6-benzo [1,3] dioxole-5-base-2-oxo-1, (1.0g is chilled to room temperature behind 10mL phosphorus chloride oxygen backflow 4h 3.2mol) to 2-dihydro-quinoline-4-formic acid 3.Solution concentration to doing, is obtained the sundown solid.Then solid is dissolved in the 20mL dichloromethane.Under 0 ℃ to solution slowly add diisopropylethylamine (1.50g, 11.5mmol) and 2-(S)-pyrrolidinylmethylpyamidesine (0.59g, 3.84mmol).At room temperature stir gained mixture 12h.After rotary evaporation removed and desolvates, residue was dissolved in the ethyl acetate, uses saturated NaHCO ₃Aqueous solution and salt water washing.The organic facies dried over sodium sulfate concentrates.Residue is by silica gel column chromatography (Et ₃N-AcOEt 5: 95) purification obtains (6-benzo [1,3] dioxole-5-base-2-chloro-quinolyl-4)-(2-pyrrolidine-1-ylmethyl-pyrrolidine-1-yl)-ketone 4 (1.20g, 81%).MSm/z?464.2(M ⁺+1)； ¹H-NMR(400MHz，CDCl ₃)：δ8.05(m，1H)，7.93(m，1H)，7.85(brs，1H)，7.41(s，1H)，7.13(m，1H)，7.11(s，1H)，6.92(m，1H)，6.03(s，1H)，4.58(m，1H)，3.92(m，0.5H)，3.75(m，0.5H)，3.38-3.13(m，2H)，2.94(m，3H)，2.35-2.15(m，2H)，2.04(m，3H)，2.01-1.85(m，4H)，1.83(m，1H)。

Under 120 ℃, with 2-chloroquinoline 4 (80mg, benzylamine 0.17mg) (0.3mL) solution heating 12h.LC-MS the analysis showed that and reacts completely.Then reactant mixture is dissolved in 3mLDMSO/CH ₃Among the CN (3: 1),, obtain product 5 by preparation type LC purification.MS?m/z?535.3(M ⁺+1)。

Synthesizing of embodiment 2:2-alkyl quinoline

Under argon atmospher, with chlorination 4-benzyl chloride base magnesium solution (0.43mL, 0.25M Et ₂O solution) join 2-chloroquinoline 4 (25mg, 0.054mg) and PdCl ₂(dppf) (2.2mg, 0.0027mmol) mixture in the 0.5mL diox.Under 100 ℃, reaction mixture is stirred 12h.After being chilled to room temperature, add NH ₄The Cl aqueous solution.The gained mixture extracts with EtOAc, uses the salt water washing.Organic layer Na ₂SO ₄Drying concentrates.Residue obtains product 6 by preparation type LC purification.MS?m/z?554.3(M ⁺+1)。

Synthesizing of embodiment 3:6-substd quinolines

Method A:

To be equipped with hypoboric acid two pinacol esters (279mg, 1.1mmol), KOAc (294,3.0mmol) and PdCl ₂(dppf) (24.5mg adds 6-iodine quinoline 7 (607mg, DMSO 1.0mmol) (6mL) solution in 25ml round-bottomed flask 0.03mmol).By flask alternately is connected with argon with vacuum, mixture is thoroughly outgased.The gained mixture is 80 ℃ of following heated overnight then, and with EtOAc (40mL) dilution, CELITE filters.Products therefrom 8 concentrates the back and uses in following steps without just being further purified.Molecular weight is 608.3, and LC-MS shows 609.2 (M ⁺+ 1).

Under argon atmospher, (15mg, 0.025mmol) De diox (2.0mL) solution joins Pd (dppf) Cl is housed with 6-borate 8 ₂(2mg), Cs ₂CO ₃(17mg, 0.055mmol) with 3, (15mg is in flask 0.057mmol) for 4-ethylenedioxy iodobenzene.By flask alternately is connected with argon with vacuum, mixture is thoroughly outgased.Then gained solution is heated to 70 ℃, stirring is spent the night.After solution is chilled to room temperature, dilute with EtOAc.Solid removes by filter by CELITE, with a little EtOAc washing.Concentrate to remove and desolvate, the gained residue obtains product 9 by the LC purification.Exact mass is 616, and LC-MS shows 617 (M ⁺+ 1).

Method B:

6-iodine quinoline 7 (25.0mg, 0.0411mmol, 1.00 equivalents), Pd (dppf) Cl pack in reaction vessel ₂(1.5mg, 0.0021,0.050 equivalent), K ₃PO ₄(35.0mg, 0.164mmol, 4.00 equivalents) and 3,5-Dichlorobenzene base boric acid (15.7mg, 0.0811mmol, 2.00 equivalents).Behind argon purge reaction vessel, under argon atmospher, introduce diox (2.0mL), the gained suspension stirs down at 80 ℃ and spends the night.Allow crude product mixture be chilled to room temperature, at the auxiliary CELITE that filters down of EtOAc, concentrated.Thick residue obtains product 10 by the LC purification.LC-MSm/z?627.2(M ⁺+1)。

Use identical method, from the synthetic 6-aryl quinoline of corresponding 6-bromoquinoline.

Method C:

To be equipped with 6-iodine quinoline 7 (61mg, 0.1mmol), phenylacetylene (1 molar equivalent), Pd (PPh ₃) ₂Cl ₂(4.0mg) and in the round-bottomed flask of CuI (1.0mg) add Et ₃N (2mL).By flask alternately is connected with argon with vacuum, mixture is thoroughly outgased, then with mixture heated to 45-50 ℃, stirring is spent the night.After mixture is chilled to room temperature,, filter CELITE with EtOAc (20ml) dilution.After concentrating, gained crude product 11 uses in next step without just being further purified.By a spot of crude product of LC purification.Exact mass is 582, and LC-MS shows 583 (M ⁺+ 1).

At room temperature and H ₂Under (1 atmospheric pressure), and 5%Pd/C (5mg) exists down in the methanol, uses H ₂Reduction crude product acetylide 11 (15mg) spend the night.Mixture filters CELITE, removes catalyst, and concentrating under reduced pressure obtains crude product.Chemical compound 12 is further purified by LC.Exact mass is 586, and LC-MS shows 587 (M ⁺+ 1).

At room temperature and H ₂Under (1 atmospheric pressure), and in methanol, in the presence of the 5%Pd/ barium sulfate (5mg), crude product acetylide 11 (15mg) reduction is spent the night.Mixture filters CELITE, removes catalyst, and concentrating under reduced pressure obtains crude product.Obtain pure products 13 by preparation type LC, be for experiment.Exact mass is 584, and LC-MS shows 585 (M ⁺+ 1).

Synthesizing of embodiment 4:2-sulfo-quinoline

Under argon atmospher, (15mg, 0.032mmol) (8.6mg adds CS in dry DMF 0.048mmol) (1.0mL) solution with 4-chloro benzyl sulfur alcohol to 2-chloroquinoline 4 ₂CO ₃(16mg, 0.048mmol).Gained mixture heated to 80 ℃, and under this temperature, stirred two hours.After being chilled to room temperature, add EtOAc (20ml) diluted mixture thing, organic facies water and salt water washing, Na ₂SO ₄Dry.After concentrating, residue obtains product 14 by the LC purification.Exact mass is 586, and LC-MS shows 587 (M ⁺+ 1).

Embodiment 5:DELFIA measures

Before starting kinase reaction, chemical compound and ZAP-70 precincubation.The precincubation reaction comprises 62.5mM Mops pH 7.0,12.5mM MgCl ₂, (glu, Tyr), 0.1mg/ml BSA, 6.25%DMSO and 0-100mM chemical compound, cumulative volume is 40ml for 12.5% glycerol, 3.1nM ZAP-70,62.5nM biotinylation poly.At room temperature incubation is after 10 minutes, and the 5mM ATP that adds 10ml starts reaction.Reacted at room temperature incubation 30 minutes, and added 5ml 500mM EDTA then and stop.With the disassociation enhancement mode lanthanide series fluorescence immunoassay instrument (DELFLA) of Perkin Elmer company, transfer to biotinylation poly (glu, phosphatic amount tyr) by the explanation measurement of manufacturer.Briefly, (glu Tyr) is captured in the Succ-PEG-DSPE bag by on the plate to biotinylation poly, uses the anti-phosphotyrosine antibody incubation of europium labelling after the washed twice.Wash and remove free antibody six times, europium is come out from the antibody disassociation, measure europium fluorescence with the excitation wavelength of 340nM and the emission wavelength of 615nM.

Table 2: the external activity of representative compounds ^*

The sequence number activity

1 B

2 B

3 A

4 C

5 B

6 A

7 A

8 A

9 A

10 A

11 B

12 C

13 C

14 A

15 C

16 C

17 C

18 C

19 C

20 A

21 A

22 C

23 C

24 C

25 C

26 A

27 C

28 A

29 B

30 C

31 C

32 A

33 B

34 B

35 C

36 C

37 C

38 B

39 B

40 C

41 C

42 B

43 C

44 C

45 C

46 C

47 B

48 B

49 B

50 C

51 C

52 C

53 C

54 C

55 C

56 C

57 C

58 B

59 B

60 C

61 B

62 B

63 B

64 B

65 C

66 B

67 A

68 C

69 A

70 C

71 A

72 B

73 B

74 B

75 B

76 C

77 C

78 B

79 B

80 A

81 B

82 B

83 B

84 A

85 B

86 B

87 A

88 A

89 B

90 B

91 C

92 C

93 A

94 B

95 B

96 B

97 B

98 C

99 C

100 A

101 B

102 B

103 B

104 B

105 C

106 C

107 C

108 B

109 B

110 A

111 A

112 A

113 A

114 A

115 A

116 B

117 C

118 B

119 A

120 B

121 B

122 B

123 B

124 B

125 C

126 C

127 C

128 A

129 B

130 B

131 B

132 B

133 B

134 B

135 B

136 B

137 B

138 A

139 B

140 C

141 B

142 B

143 B

144 B

145 B

146 B

147 B

148 B

149 B

150 A

151 A

152 A

153 A

154 A

155 A

156 A

157 A

158 A

159 A

160 A

161 A

162 A

163 A

164 A

165 A

166 A

167 A

168 A

169 A

170 A

171 A

172 A

173 A

174 A

175 A

176 A

177 A

178 A

179 A

180 A

181 A

182 A

183 A

184 A

185 A

186 A

187 A

188 A

189 A

190 A

191 A

192 A

193 A

194 A

195 A

196 A

197 A

198 A

199 A

200 A

201 A

202 A

203 A

204 A

205 A

206 A

207 A

208 A

209 A

210 A

211 A

212 A

213 A

214 A

215 A

216 A

217 A

218 A

219 A

220 A

221 A

222 A

223 A

224 A

225 A

226 A

227 A

228 A

229 A

230 A

231 A

232 A

233 A

234 A

235 A

236 A

237 A

238 A

239 A

240 A

241 A

242 A

243 A

244 A

245 A

246 A

247 A

248 A

249 A

250 A

251 A

252 A

253 A

254 A

255 A

256 A

257 A

258 A

259 A

260 A

261 A

262 A

263 A

264 A

265 A

266 A

267 A

268 A

269 A

270 A

271 A

272 A

273 A

274 A

275 A

276 A

277 A

278 A

279 A

280 C

281 C

282 B

283 C

284 C

285 B

286 C

287 B

288 B

289 B

290 B

291 A

292 C

293 B

294 C

295 B

296 B

297 A

298 B

299 C

300 C

301 1.785μM

302 C

303 C

304 B

305 C

306 C

307 B

308 B

309 B

310 C

311 B

312 B

313 B

314 B

315 C

316 B

317 C

318 B

319 C

320 B

321 B

322 -

323 A

324 C

325 C

326 C

327 A

328 B

329 A

330 C

331 C

332 A

333 A

334 A

335 A

336 A

337 A

338 A

339 A

340 A

341 A

342 A

343 A

344 C

345 A

346 A

347 A

348 C

349 C

350 A

351 C

352 C

353 A

354 A

355 A

356 A

357 B

358 C

359 C

360 B

361 B

362 A

363 B

364 B

365 B

366 B

367 A

368 A

369 A

370 A

371 A

372 B

373 B

374 B

375 B

376 B

377 A

378 A

379 A

380 A

381 A

382 A

383 A

384 C

385 B

386 B

387 B

388 B

389 C

390 A

391 A

392 C

393 C

394 A

395 C

396 C

397 C

398 C

399 A

400 C

401 C

402 C

403 C

404 C

405 C

406 C

407 C

408 A

409 A

410 C

411 C

412 A

413 B

414 A

415 A

416 A

417 A

418 A

419 A

420 A

421 A

422 A

423 A

424 A

425 A

426 A

427 A

428 C

429 C

430 C

431 B

432 C

433 C

434 C

435 C

436 C

437 C

438 C

439 C

440 C

441 C

442 C

443 C

444 C

445 C

446 C

447 C

448 C

449 C

^*" A " refers to the external IC of chemical compound ₅₀＞10 μ M, " B " refer to the external IC of chemical compound ₅₀Between 1-10 μ M, " C " refers to the external IC of chemical compound ₅₀＜1 μ M.

The mensuration based on cell of embodiment 6:ZAP-70 mediation LAT phosphorylation

ZAP-70 tyrosine kinase activity in the activation Jurkat T cell is measured by the phosphorylation state of assessment LAT.LAT is the palmitoylation p36/38kDa plasma membrane associated protein that is expressed in all T lymphocytes, is the physiological substrate of activation ZAP-70.When the Jutkat cell stimulates with the CD 3-resisting monoclonal antibody of crosslinked surface receptor antigen-CD3 complex, ZAP-70 be activated and phosphorylation LAT on two tyrosine residue Tyr-191 and Tyr-226.So the LAT Tyr-191 in the anti-CD3 stimulation Jurkat cell and the assessment of Tyr-226 phosphorylation are to the active concrete standard of ZAP-70 in the born of the same parents.Therefore, the compound exhibits of inhibition LAT Tyr-191 and Tyr-226 phosphorylation goes out the activatory successful antagonism of ZAP-70 tyrosine kinase activity or ZAP-70 approach upstream.

With 15-20 * 10 ⁶The Jurkat cell (ATCC) that the concentration of individual cell/mL is cultivated and inhibitor compound or carrier (DMSO) are CD 3-resisting monoclonal antibody (UCHT-1 or the OKT-3 of 1mg/mL by adding final concentration 37 ℃ of following preincubates 15 minutes then; E-bioscience) stimulated 5 minutes.Then, by buffer cell lysis, use commercially available phosphotyrosine monoclonal antibody specific (4G10 based on detergent; Upstate Biotechnology Inc.) and A albumen conjugation agarose beads, the protein to all the phosphoric acid tyrosine in the extract carries out immunoprecipitation.The washing immunocomplex with the protein of removing non-specific binding after, by boiling the phosphotyrosine protein of immunoprecipitation is discharged from agarose beads with degeneration, separate with SDS-PAGE, and transfer on the nitrocellulose filter.Then, western blotting by standard, use can with the phosphoric acid Tyr-191 on the LAT or commercially available phosphoric acid specific antibody (Upstate Biotechnology Inc.), enzyme conjugation second antibody (horseradish peroxidase conjugation goat anti-rabbit igg antiserum) and the Storm imaging system (Amersham-Pharmacia) of phosphoric acid Tyr-226 specific reaction, detect on Tyr-191 and Tyr-226 by the LAT of phosphorylation.

Observed inhibition with representative compounds described herein.

The mensuration that embodiment 7:IL-2 suppresses based on cell

After the Jurkat cell stimulated altogether with anti-CD3 and anti-CD28 antibody, the IL-2 that measures wherein generated situation.Cell is with 1 * 10 ⁵Individual/hole (culture 200 μ l/ holes) are inoculated in pre-bag by 96 orifice plates of anti-cd 3 antibodies (BD Biocoat T-cell activation plate, anti-people CD3 plate=catalogue No 354725, BD BioSciences).To resist CD28 antibody (eBioScience, catalogue No 16-0289-85; Functional level stimulates altogether, clone 28.2) join in the plate to final concentration be the 20ng/ hole.After cell was hatched 48 hours, the five equilibrium cell conditioned medium liquid of drawing 50 μ l carried out the IL-2 titration.The IL-2 titration is undertaken by manufacturer's explanation with Endogen enzyme-linked immunosorbent assay (ELISA) test kit (Endogen ofPierce, catalogue No EH2-IL2-5).Briefly, this measures by using pre-bag to be formed by 96 hole microtitration plates of anti-IL-2 antibody, in order to catch the human IL-2.IL-2 for detection is captured joins the biotinylated second anti-IL-2 antibody in all holes.This causes sandwichization (catch IL-2 antibody C IL-2C and detect the IL-2-biotinylated antibody) of any IL-2.After removing unconjugated antibody by a series of washing, add biology is have high-affinity horseradish peroxidase (HRP) conjugate.Remove unconjugated Succ-PEG-DSPE-HRP, bonded enzymic-labelled antibody just can be measured by adding tmb substrate generation chromogenic reaction.The chromogenic reaction H that causes ₂SO ₄(stop buffer) stops, then the optical density in each hole reading under suitable wavelength.The level of conversion of substrate is carried out colorimetric determination by measuring absorbance, and it is directly proportional with the amount of IL-2.

Representative compounds described herein shows that IL-2 suppresses.

All lists of references that this paper quotes, no matter occur with printed article, electronic medium, computer readable storage medium or other forms, all obviously by reference integral body be attached to herein, described list of references includes but not limited to summary, article, periodical, publication, textbook, paper, internet website, data base, patent and patent publications.

A plurality of embodiment of the present invention has been described.But should be understood that and not break away from the spirit and scope of the present invention, make different improvement projects.Therefore, other embodiments are also included in the scope of following claims.

Claims

1. a formula (I) chemical compound

Formula (I)

Wherein,

N is 0-3;

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁸Independently be OR ⁵Or alkyl;

2. the chemical compound of claim 1, described chemical compound has formula (I), wherein

A forms phenyl ring.

3. the chemical compound of claim 1, described chemical compound has formula (II)

Formula (II)

Wherein,

Each R ²Independently be H, halogen, NH ₂, alkyl, OH, C (O) Me, aryl, heteroaryl;

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁸Independently be OR ⁵Or alkyl;

Het is optional by 1-4 R ¹⁰The heterocyclic radical that replaces;

4. the chemical compound of claim 3, described chemical compound has formula (II), wherein

Het connects by theheterocyclic nitrogen atom.

5. the chemical compound of claim 3, described chemical compound has formula (II), wherein

R ²' be H.

6. the chemical compound of claim 3, described chemical compound has formula (II), wherein;

Het connects by theheterocyclic nitrogen atom;

X is NR ³Or alkyl;

R ^XBe cycloalkyl, aryl, heteroaryl, aralkyl, heteroarylalkyl or heterocyclic radical, wherein R ^XOptional by 1-4 R ⁴Replace;

R ^{2 '}Be H.

7. the chemical compound of claim 3, described chemical compound has formula (II), wherein

Het is

Or

8. the chemical compound of claim 3, described chemical compound has formula (II), and wherein Het is

9. the chemical compound of claim 1, described chemical compound has formula (III),

Formula (III)

Wherein,

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁸Independently be OR ⁵Or alkyl;

10. the chemical compound of claim 9, described chemical compound has formula (III), wherein R ^{2 '}Be H.

11. the chemical compound of claim 9, described chemical compound have formula (III), wherein

R ¹²Be

Or

A 12. formula (IV) chemical compound

Formula (IV)

Wherein,

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁷Independently be halogen, CN, OR ⁵, CF ₃, N (R ⁵) C (O) R ⁵, C (O) R ⁵, OCF ₃, SCF ₃, NR ⁵ ₂, C (O) NR ⁵ ₂, OH, R ₅O-alkyl, alkyl, alkyl sulphonyl, heterocyclic radical or heteroaryl, they are optional separately by 1-4 R ⁸Replace;

Each R ⁸Independently be OR ⁵Or alkyl;

13. formula V chemical compound

Formula V

Wherein,

Each R ²Independently be H, NR ⁵ ₂, alkyl, cycloalkyl, thiazolinyl, alkynyl, heteroaryl, heterocyclic radical, aryl, halogen, acetyl group, aralkyl, heteroarylalkyl, aryl alkenyl, heteroaryl thiazolinyl, aromatic yl polysulfide yl, heteroaryl alkynyl, hydroxyl, alkoxyl, aryloxy group or heteroaryloxy, they are optional separately by 1-4 R ⁷Replace; Wherein work as R ^YWhen the indole that is halogen, is replaced by halogen, phenyl or the phenyl that replaced by halogen, R ²Not Me; Wherein work as R ^YDuring the phenyl that replaced by halogen, R ²It or not isopropyl; Wherein work as R ^YDuring the indole that replaced by halogen, R ²The alkynyl that is replaced by heterocyclic radical not; Wherein work as R ^YWhen being H, R ²It or not phenyl; Wherein work as R ^YWhen indole that is replaced by halogen or the phenyl that replaced by halogen, R ²It or not halogen; And wherein work as R ^YWhen being phenyl or substituted-phenyl, R ²It or not acetyl group;

N is 0-3;

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁸Independently be OR ⁵Or alkyl;

14. the chemical compound of claim 13, described chemical compound has formula V, wherein

A forms phenyl ring.

15. the chemical compound of claim 13, described chemical compound have formula (VI), wherein

Formula (VI)

Wherein,

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁸Independently be OR ⁵Or alkyl;

Het is optional by 1-4 R ¹⁰The heterocyclic radical that replaces;

16. the chemical compound of claim 15, described chemical compound have formula (VI), wherein

Het connects by theheterocyclic nitrogen atom;

R ²Not H;

R ^{2 '}Be H.

17. the chemical compound of claim 15, described chemical compound have formula (VI), wherein

Het is

Or

18. the chemical compound of claim 15, described chemical compound have formula (VI), wherein Het is

19. the chemical compound of claim 13, described chemical compound have formula (VII)

Formula (VII)

Wherein,

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁸Independently be OR ⁵Or alkyl;

20. the chemical compound of claim 19, described chemical compound have formula (VII), wherein

R ²Be not ^H

R ^{2 '}Be H.

21. the chemical compound of claim 19, described chemical compound have formula (VII), wherein

R ¹²Be

Or

22. a formula (VIII) chemical compound,

Formula (VIII)

Wherein,

Each n is 0-3;

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁸Independently be OR ⁵Or alkyl;

23. the chemical compound of claim 22, described chemical compound have formula (VIII), wherein n is 1, R ²Not H.

24. a formula (XIV) chemical compound,

Formula (XIV)

Wherein,

N is 0-3;

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁸Independently be OR ⁵Or alkyl;

25. the method for a preparation formula (II) chemical compound

Formula (II)

Wherein X, R ^X, R ², R ²' being defined as follows with Het, described method comprises;

Formula (IX) formula (X)

Formula (X) formula (XI) formula (XII)

Use POCl ₃Processing formula (XII) chemical compound obtains formula (XIII) chloride, and makes the amine coupling of formula (XII) carboxylic acid and formula Het-H, obtains formula (XIII) chemical compound;

Formula (XII) formula (XIII)

Formula (XIII) formula (II)

Wherein for formula II and formula IX-XIII,

R ^{2 '}Be H, halogen, NH ₂, alkyl, OH, C (O) Me, aryl, heteroaryl;

Each R ³Independently be H, alkyl, R ⁵O-alkyl or aralkyl;

Each R ⁵Independently be H or alkyl;

Each R ⁸Independently be OR ⁵Or alkyl;

Het is optional by 1-4 R ¹⁰The heterocyclic radical that replaces;

26. the method for claim 25, wherein the H of Het-H is connected with nitrogen.

27. the method for claim 25, wherein coupling agent is H ₂NR ^X

28. the method for claim 25, wherein coupling agent is MgCl-R ^XAnd Pd.

29. the method for claim 25, wherein coupling agent is HSR ^X

30. comprising, a method for the treatment of autoimmune disease among the patient, described method give patient's claim 1-24 each chemical compound.

31. further comprising, the method for claim 30, described method give other medicine.

32. the method for claim 30, wherein autoimmune disease is a lupus.

33. comprising, a method for the treatment of organ-graft refection among the patient, described method give patient's claim 1-24 each chemical compound.

34. further comprising, the method for claim 33, described method give other medicine.

35. comprising, a method for the treatment of inflammatory diseases among the patient, described method give patient's claim 1-24 each chemical compound.

36. further comprising, the method for claim 35, described method give other medicine.

37. the method for claim 36, wherein said other medicine is analgesic or steroid.

38. the method for claim 35, wherein said inflammatory diseases is an arthritis.

39. the method for claim 38, wherein said arthritis are rheumatoid arthritis, rheumatoid spondylitis, gouty arthritis, traumatic arthritis, rubella arthritis, psoriatic arthritis or osteoarthritis.

40. the method for claim 35, wherein said inflammatory diseases are inflammatory bowel or Crohn disease.

41. a compositions, described compositions comprise each chemical compound of claim 1-24.

42. the compositions of claim 41, described compositions further comprises drug acceptable carrier.

43. the compositions of claim 41, described compositions further comprises other medicine.

44. a compound library, described chemical compound are the chemical compound of any formula (I)-(VIII).

45. one kind is suppressed the method that IL-2 among the patient generates, described method comprises and gives patient's claim 1-24 each chemical compound.

46. comprising, a method of regulating ZAP-70 among the patient, described method give patient's claim 1-24 each chemical compound.