CN101663297B - Oxadiazole substituted indazole derivatives for use as sphingosine 1-phosphate (s1p) agonists - Google Patents

Oxadiazole substituted indazole derivatives for use as sphingosine 1-phosphate (s1p) agonists Download PDF

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CN101663297B
CN101663297B CN200880012733.XA CN200880012733A CN101663297B CN 101663297 B CN101663297 B CN 101663297B CN 200880012733 A CN200880012733 A CN 200880012733A CN 101663297 B CN101663297 B CN 101663297B
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indazole
phenyl
diazole
oxygen base
compound
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CN101663297A (en
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马穆德·马梅德
杰勒德·M·P·吉布林
詹姆斯·米亚特
戴维·诺顿
迪安·A·里弗斯
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Glaxo Group Ltd
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Abstract

The present invention provides compounds of formula (I) or salts thereof, having pharmacological activity, processes for their preparation, pharmaceutical compositions containing them and their use in the treatment of various disorders mediated by S1P1 receptors.

Description

Be used as the indazole derivatives of the * diazole replacement of S1P (S1P) agonist
The present invention relates to have the Xin De oxadiazole derivative of pharmacological activity, their preparation method contains their pharmaceutical composition and they are in the purposes for the treatment of in various diseases.
S1P (S1P) is a kind of biological activity lipoid amboceptor that is formed by Sphingosine kinase phosphorylation sphingosine, and finds that it is higher at blood middle concentration.Many cell types are all produced and are secreted S1P, comprise cell (Okamoto etc., 1998 JBiol Chem 273 (42): 27104 as those hematopoietic origins such as thrombocyte and mastocyte; Sanchez and Hla 2004, J Cell Biochem 92:913).Its biological action is wider, comprises the adjusting of on cell proliferation, differentiation, mobility, vascularization and to inflammatory cell and hematoblastic activation (Pyne and Pyne 2000, Biochem J.349:385).Record at present the S1P response acceptor of five kinds of hypotypes, S1P1 (Edg-1), S1P2 (Edg-5), S1P3 (Edg-3), S1P4 (Edg-6) and S1P5 (Edg-8), they have formed a part (Chun of endothelial differentiation gene family of the G-protein coupling of acceptor, Deng, 2002 Pharmacological Reviews 54:265, Sanchez and Hla2004 J Cellular Biochemistry, 92:913).These 5 kinds of acceptors show different mrna expressions, and wherein S1P1-3 is by wide expression, and S1P4 is expressed on Lymphoid tissue and hemopoietic tissue, S1P5 mainly express in brain and than low degree ground expression in spleen.They send signal by the different subclass of G protein, to facilitate various biological responses (Kluk and Hla, 2002 Biochem et Biophysica Acta1582:72, Sanchez and Hla 2004, J Cellular Biochem 92:913).
The effect of the S1P1 acceptor that proposes comprises the transportation lymphocyte, induces/suppress cytokine, and the effect of Human Umbilical Vein Endothelial Cells (Rosen and Goetzl, 2005 Nat Rev Immunol.5:560).The agonist of S1P1 acceptor has been used in the animal model of many autoimmunizatioies and transplanting, comprises tentative autoimmunization encephalomyelitis (EAE) model of MS, induces seriousness (the 2003 JBC 277:21453 such as Brinkman of disease with minimizing; The 2003 J Pharmacol Exp Ther 305:70 such as Fujino; The 2004 J Neuroimmunol 153:108 such as Webb; The 2004 J Magn Reson Imaging20:16 such as Rausch).Report thinks that the effect through lymphoid circulation mediates this activity to lymphocyte by the S1P1 agonist.Treat with the S1P1 agonist and produced the second lymphoid organ such as the lymphocytic isolation in lymphoglandula inside (sequestration), comprise that reversible periphery lymphocyte in animal model generates that (Chiba etc. 1998, J Immunology 160:5037, the 2004 J Pharmacol ExpTher 309:758 such as Forrest; The 2004 JBC 279:13839 such as Sanna).About agonist, published data show that compounds for treating induced the S1P1 acceptor from loss (Graler and the Goetzl2004FASEB J 18:551 of cell surface because of internalization; The 2004 Nature 427:355 such as Matloubian; The 2005 ChemBiol 12:703 such as Jo), and the minimizing of this S1P1 acceptor on immunocyte help to reduce the T cell and get back to blood flow from lymphoglandula motion.
S1P1 genetically deficient has caused the embryonic death phenomenon.The experiment that checks the effect of S1P1 acceptor in lymphocyte migration and transportation has comprised the adoptive transfer of the wild-type mice of T cell after the radiation of the S1P1 disappearance of mark.These cells show the outflow minimizing (the 2004 Nature 427:355 such as Matloubian) from the second lymphoid organ.
S1P1 also is considered to have the effect that endotheliocyte connect to regulate people 2003102:3665 such as (, the people 2005 FASEB J 19:1646 such as Blood Singelton) Allende.About this endothelium effect, reported the S1P1 agonist isolated lymph is had effect, and solitary lymph follicle may there be regulating effect to immune disorders.The S1P1 agonist has caused closing of matter " door " between the endothelium of lymphatic sinus, and emptying (drain) lymphoglandula of matter " door " and prevent that lymphocyte from flowing out (Wei wtal 2005, Nat.Immunology 6:1228) between the endothelium of described lymphatic sinus.
immunosuppressive compounds FTY720 (JP11080026-A) has shown the circulating lymphocyte that reduces in animal and human's body, having disease in the animal model of immune disorders regulates active, and alleviation speed (the 2002 JBC 277:21453 such as Brinkman in the multiple sclerosis of alleviating recurrence (relapsing remitting) have been reduced, the 2002 Science 296:346 such as Mandala, the 2003 JPharmacology and Experimental Therapeutics 305:45658 such as Fujino, the 2004American J Transplantation 4:1019 such as Brinkman, the 2004 J Neuroimmunology 153:108 such as Webb, the 2005 EurJ Immunol 35:3570 such as Morris, Chiba 2005 Pharmacology andTherapeutics 108:308, Kahan etc. 2003, Transplantation 76:1079, the 2006New Eng J Medicine 335:1124 such as Kappos).This compound is a kind of prodrug, and it is obtained S1P1, S1P3, S1P4 and S1P5 acceptor are had the molecule of agonist activity in vivo by the Sphingosine kinase phosphorylation.Clinical study shows with FTY720 treatment cause bradyrhythmia (the 2006 New Eng J Medicine 335:1124 such as Kappos) in 24 hour for the treatment of.According to many experiments based on cell and experimentation on animals, think that this bradyrhythmia is because of the agonism to S1P3.These experiments comprise uses the S1P3 knock-out animal that is different from wild-type mice, and it does not demonstrate bradyrhythmia at administration FTY720 with after using the S1P1 alternative cpd.(the 2004 Bioorganic ﹠amp such as Hale; Medicinal Chemistry Letters14:3501, the 2004JBC 279:13839 such as Sanna, the 2005 American JTransplantation 5:529 such as Koyrakh).
Following patent application is described oxadiazole derivative as S1P1 agonist: WO03/105771, WO05/058848, WO06/047195, WO06/100633, WO06/115188 and WO06/131336.
Following patent application has been described indoles-oxadiazole derivatives as anti-pico+ribonucleic acid+virus medicine (antipicornaviral agents): WO96/009822.Following patent application has been described the indole-2-formate derivative respectively as leukotrienes receptor antagonist, sterilant and agricultural bactericide: WO06/090817, EP 0 439 785 and DE 39 39 238.
International Patent Application WO 06/001463 discloses multiple compound as the S1P1 receptor stimulant.
International Patent Application PCT/EP2007/064185 discloses indoles-oxadiazole compounds as the S1P1 receptor stimulant.
Have now found that the compound of a class new texture, it provides the agonist of S1P1 acceptor.
Therefore, the invention provides formula (I) compound or its salt:
Figure G200880012733XD00031
Wherein
R 5And R 6In one be hydrogen or R 2, another is (a)
R 3And R 4One of be (b)
Figure G200880012733XD00033
A is phenyl or 5 or 6-unit hetero-aromatic ring;
R 1Be hydrogen or 3 substituting groups at the most, described substituting group is independently selected from: halogen, C (1-4)Alkyl, C (1-4)Alkoxyl group, C (5-7)Cycloalkyl, C (5-7)Cycloalkyloxy, trifluoromethoxy, difluoro-methoxy, trifluoromethyl, cyano group, phenyl, 5 or 6 yuan of hetero-aromatic rings, piperidyl and pyrrolidyls;
R 2Be hydrogen or 3 substituting groups at the most, described substituting group is independently selected from: halogen, C (1-4)Alkyl, C (1-4)Alkoxyl group, trifluoromethoxy, difluoro-methoxy, trifluoromethyl and cyano group;
R 7Be hydrogen or halogen;
Z is C (1-4)Alkyl, it can choose N of insertion (interrupt) or O wantonly, and it can be chosen wantonly on carbon by 4 substituting groups at the most and replace, and described substituting group is independently selected from: halogen, methyl and hydroxyl, condition are not have carbon atom to be replaced by two hydroxyls.
Therefore, the invention provides formula (IA) compound or its salt:
Figure G200880012733XD00041
Wherein A, R 1, R 2, R 5, R 6, R 7Define suc as formula (I) with Z.
Therefore, the invention provides formula (IB) compound or its salt:
Figure G200880012733XD00042
Wherein A, R 1, R 2, R 5, R 6, R 7Define suc as formula (I) with Z.
Therefore, the invention provides formula (II) compound or its pharmaceutically acceptable derivates:
Figure G200880012733XD00043
Wherein
R 5And R 6In one be hydrogen or R 2, another is (a)
Figure G200880012733XD00044
R 3And R 4One of be hydrogen, and another is (b)
Figure G200880012733XD00051
A is phenyl or 5 or 6-unit hetero-aromatic ring;
R 1Be hydrogen or 3 substituting groups at the most, described substituting group is independently selected from: halogen, C (1-4)Alkyl, C (1-4)Alkoxyl group, trifluoromethoxy, difluoro-methoxy, trifluoromethyl, cyano group, and optional substituted phenyl or optional substituted 5 or 6 yuan of hetero-aromatic rings;
R 2Be hydrogen or 3 substituting groups at the most, described substituting group is independently selected from: halogen, C (1-4)Alkyl, C (1-4)Alkoxyl group, trifluoromethoxy, difluoro-methoxy, trifluoromethyl and cyano group;
R 7Be hydrogen or halogen;
Z is C (1-4)Alkyl, its optional N or O of inserting, and optional by halogen or methyl substituted.
Work as R 1During for phenyl or 5 or 6 yuan of hetero-aromatic rings, they can be by 3 substituting groups replacements at the most, and described substituting group is selected from: halogen, C (1-4)Alkyl, C (1-4)Alkoxyl group, trifluoromethoxy, difluoro-methoxy, trifluoromethyl and cyano group.
Should be understood that for formula (II) compound, work as R 4During for (b), R 3Do not exist, and arrange described in the chemical bond in aromatic nucleus such as above-mentioned formula (IB).
Term " alkyl " refers to the straight or branched alkyl of all isomeric form as the part of a group or group (as alkoxyl group or hydroxyalkyl).Term " C (1-4)Alkyl " refer to alkyl as defined above, it contains at least 1 and 4 carbon atoms at the most.The example of such alkyl comprises methyl, ethyl, propyl group, sec.-propyl, normal-butyl, isobutyl-, sec-butyl, the tertiary butyl, 1-methyl-propyl or 2-methyl-propyl.The example of such alkoxyl group comprises methoxyl group, oxyethyl group, propoxy-, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert.-butoxy or 1-methyl propoxy-.
The C that is fit to (5-7)Cycloalkyl comprises cyclopentyl, cyclohexyl and suberyl.
The C that is fit to (5-7)Cycloalkyloxy comprises cyclopentyloxy, cyclohexyloxy and ring oxygen in heptan base.
Term used herein " halogen " refers to fluorine (F), chlorine (Cl), bromine (Br) or iodine (I), and term " halo " refers to halogen: fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).
Term " heteroaryl " expression comprises one or more heteroatomic unsaturated rings.When this term heteroaryl represented 5-unit group, it comprised the heteroatoms that is selected from O, N or S, and can choose wantonly and contain 1-3 nitrogen-atoms again.When heteroaryl represented the first group of 6-, it comprised 1-3 nitrogen-atoms.The example of 5 or 6 yuan of hetero-aromatic rings like this comprises pyrryl, triazolyl, thiadiazolyl group, tetrazyl, imidazolyl, pyrazolyl, isothiazolyl, thiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, furazan base, furyl, thienyl, pyridyl, pyrimidyl, pyrazinyl, pyridazinyl and triazinyl.
One embodiment of the invention are formula (I) compound, wherein
A is thiophene, pyridyl or phenyl; And
R 1Be two substituting groups, described substituting group is independently selected from: chlorine, bromine, isopropoxy, propoxy-, methoxyl group, 1-methyl propoxy-, cyano group, trifluoromethyl, trifluoromethoxy, cyclohexyl, piperidines, pyrrolidyl, ethyl, 2-methyl-propyl, phenyl and cyclopentyloxy; And
R 2Be hydrogen; And
R 7Be hydrogen; And
Z is ethylidene or propylidene, is optionally separately replaced together with-dimethyl.
One embodiment of the invention are formula (I) compound, wherein
R 6Be (a), and R 5Be hydrogen; And
A is thiophene, pyridyl or phenyl; And
R 1Be two substituting groups, described substituting group is independently selected from: chlorine, bromine, isopropoxy, propoxy-, methoxyl group, 1-methyl propoxy-, cyano group, trifluoromethyl, trifluoromethoxy, cyclohexyl, piperidines, pyrrolidyl, ethyl, 2-methyl-propyl, phenyl and cyclopentyloxy; And
R 2Be hydrogen; And
R 7Be hydrogen; And
Z is ethylidene or propylidene, is optionally separately replaced together with-dimethyl.
One embodiment of the invention are formula (IA) compound, wherein
R 6Be (a), and R 5Be hydrogen; And
A is phenyl; And
R 1Be two substituting groups, described substituting group is independently selected from: chlorine, isopropoxy, cyano group, trifluoromethyl, trifluoromethoxy, piperidines, ethyl and phenyl; And
R 2Be hydrogen; And
R 7Be hydrogen; And
Z is ethylidene or propylidene.
One embodiment of the invention are formula (IA) compound, wherein
R 6Be (a), and R 5Be hydrogen; And
A is phenyl; And
R 1Be chlorine and isopropoxy; And
R 2Be hydrogen; And
R 7Be hydrogen; And
Z is ethylidene or propylidene.
One embodiment of the invention are formula (IB) compound, wherein
R 6Be (a), and R 5Be hydrogen; And
A is phenyl; And
R 1Be two substituting groups, described substituting group is independently selected from: chlorine, isopropoxy and cyano group; And
R 2Be hydrogen; And
R 7Be hydrogen; And
Z is ethylidene or propylidene.
One embodiment of the invention are formula (I) compound, wherein
R 5Be (a), and R 6Be hydrogen; And
A is thiophene, pyridyl or phenyl; And
R 1Be two substituting groups, described substituting group is independently selected from: chlorine, bromine, isopropoxy, propoxy-, methoxyl group, 1-methyl propoxy-, cyano group, trifluoromethyl, trifluoromethoxy, cyclohexyl, piperidines, pyrrolidyl, ethyl, 2-methyl-propyl, phenyl and cyclopentyloxy; And
R 2Be hydrogen; And
R 7Be hydrogen; And
Z is ethylidene or propylidene, is optionally separately replaced together with-dimethyl.
One embodiment of the invention are formula (IA) compound, wherein
R 5Be (a), and R 6Be hydrogen; And
A is phenyl; And
R 1Be chlorine and isopropoxy; And
R 2Be hydrogen; And
R 7Be hydrogen; And
Z is ethylidene.
One embodiment of the invention are formula (IB) compound, wherein
R 5Be (a), and R 6Be hydrogen; And/or
A is phenyl; And
R 1Be two substituting groups, described substituting group is independently selected from: chlorine, isopropoxy, phenyl and trifluoromethyl; And
R 2Be hydrogen; And
R 7Be hydrogen; And
Z is propylidene.
One embodiment of the invention are formula (II) compound, wherein
R 3Be (b); And/or
R 5Be (a), and R 6Be hydrogen; And/or
A is optional substituted thiophene or phenyl; And/or
R 1Be hydrogen, halogen, C 1-4Alkoxyl group or trifluoromethyl; And/or
R 2Be hydrogen; And/or
Z is ethylidene.
Another embodiment of the invention is formula (II) compound, wherein
R 3Be (b); And/or
R 5Be (a), and R 6Be hydrogen; And/or
A is by the thiophene of phenyl substituted; And/or
R 1Be hydrogen, halogen, C 1-4Alkoxyl group or trifluoromethyl; And/or
R 2Be hydrogen; And/or
Z is ethylidene.
The present invention includes all optically active isomers, for example the steric isomer of formula (I) compound comprises enantiomer, diastereomer and composition thereof, as racemic modification.Different steric isomers can separate with another one by ordinary method or split, or any given isomer can be by conventional three-dimensional the selection or the asymmetric synthesis acquisition.
Suitable compound of the present invention is:
3-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] propionic acid
3-(5-{5-[3-chloro-4-(oxyethyl group) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) propionic acid
3-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] propionic acid
3-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] propionic acid
4-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] butyric acid
3-(5-{5-[4-[(1-methylethyl) oxygen base]-3-(trifluoromethyl) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) propionic acid
4-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] butyric acid
4-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] butyric acid
3-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] propionic acid
3-(5-{5-[3-chloro-4-(propoxy-) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) propionic acid
3-[5-(5-{3-chloro-4-[(trifluoromethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] propionic acid
3-(5-{5-[4-cyclohexyl-3-(trifluoromethyl) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) propionic acid
3-(5-{5-[4-(methoxyl group)-3-(trifluoromethyl) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) propionic acid
[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] acetic acid
3-(5-{5-[3-chloro-4-(methoxyl group) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) propionic acid
3-(5-{5-[3-ethyl-4-(piperidino) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) propionic acid
3-{5-[5-(4-cyclohexyl-3-ethylphenyl)-1,2,4-oxadiazole-3-yl]-1H-indazole-1-yl } propionic acid
3-(5-{5-[3-cyano group-4-(2-methyl-propyl) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) propionic acid
3-[5-(5-{3-bromo-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] propionic acid
3-(5-{5-[3-chloro-4-(2-methyl-propyl) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) propionic acid
4-(5-{5-[4-[(1-methylethyl) oxygen base]-3-(trifluoromethyl) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) butyric acid
3-{5-[5-(2-cyano group-4-xenyl)-1,2,4-oxadiazole-3-yl]-1H-indazole-1-yl } propionic acid
3-(5-{5-[3-chloro-4-(1-pyrrolidyl) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) propionic acid
4-[5-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] the butyrates hydrochlorate
4-[5-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-oxadiazole-3-yl)-2H-indazole-2-yl] the butyrates hydrochlorate
4-(5-{5-[2-(trifluoromethyl)-4-xenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) butyric acid
4-(5-{5-[2-(trifluoromethyl)-4-xenyl]-1,2,4-oxadiazole-3-yl }-2H-indazole-2-yl) butyric acid
3-[4-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] propionic acid
3-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-2H-indazole-2-yl] propionic acid
3-[4-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-2H-indazole-2-yl] propionic acid
4-[4-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] butyric acid
4-(4-{5-[3-ethyl-4-(piperidino) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) butyric acid
4-(4-{5-[2-(trifluoromethyl)-4-xenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) butyric acid
4-[4-(5-{3-chloro-4-[(trifluoromethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] butyric acid
3-(4-{5-[4-[(1-methylethyl) oxygen base]-3-(trifluoromethyl) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) propionic acid
4-[4-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] butyric acid
4-(4-{5-[4-phenyl-5-(trifluoromethyl)-2-thienyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) butyric acid
4-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-2H-indazole-2-yl] butyric acid
3-{5-[5-(3-cyano group-4-{[(1S)-1-methyl-propyl] the oxygen base } phenyl)-1,2,4-oxadiazole-3-yl]-1H-indazole-1-yl } propionic acid sodium salt
3-{5-[5-(3-cyano group-4-{[(1R)-1-methyl-propyl] the oxygen base } phenyl)-1,2,4-oxadiazole-3-yl]-1H-indazole-1-yl } propionic acid sodium salt
3-(5-{5-[3-cyano group-4-(cyclopentyloxy) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) propionic acid
4-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] butyric acid
4-(5-{5-[4-phenyl-5-(trifluoromethyl)-2-thienyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) butyric acid
4-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-2H-indazole-2-yl] butyric acid
3-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-PA
3-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-2H-indazole-2-yl]-PA
3-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-PA
3-[4-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-PA
3-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-PA
4-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-2, the 2-acid dimethyl
3-[4-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-PA
3-[5-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-PA
4-[4-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-2, the 2-acid dimethyl
4-[4-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-2, the 2-acid dimethyl
Or its salt.
The pharmaceutically acceptable derivates of formula (I) compound comprises the salt of any pharmacy acceptable salt, ester or this ester of formula (I) compound, when can (directly or indirectly) provide formula (I) compound or its active metabolite or resistates after recipient's administration.
Some esters of formula (I) compound are described as the intermediate of more synthetic described embodiment in this article.Such ester also can show the activity as the S1P1 agonist, and consists of thus a part of the present invention.
Formula (I) compound can form salt.The salt that should understand application formula (I) compound in medicine should be pharmaceutically acceptable.The pharmacy acceptable salt that is fit to will be obvious for those skilled in the art, and comprise and be described in J.Pharm.Sci., 1977,66, those in 1-19, the acid salt that for example forms with mineral acid (example hydrochloric acid, Hydrogen bromide, sulfuric acid, nitric acid or phosphoric acid); Acid salt with organic acid (as succsinic acid, toxilic acid, acetic acid, fumaric acid, citric acid, tartrate, phenylformic acid, tosic acid, methylsulfonic acid or naphthene sulfonic acid) formation.Some formulas (I) compound can form acid salt with the acid of monovalent or more equivalents.The present invention comprises all possible stoichiometry and non-stoichiometric forms in its scope.Salt also can be from the acceptable alkali preparation of pharmacy, and described alkali comprises mineral alkali and organic bases.The salt that is derived from mineral alkali comprises aluminium salt, ammonium salt, calcium salt, mantoquita, molysite, ferrous salt, lithium salts, magnesium salts, manganic salt, manganous salt, sylvite, sodium salt, zinc salt etc.The salt that is derived from the acceptable organic bases of pharmacy comprises primary amine, secondary amine and tertiary ammonium salt; The amine salt that replaces comprises naturally occurring replacement amine salt; With cyclammonium salt.specific pharmaceutically acceptable organic bases comprises arginine, trimethyl-glycine, caffeine, choline, N, N '-dibenzyl-ethylenediamin, diethylamide, the 2-DEAE diethylaminoethanol, DMAE, thanomin, quadrol, N-ethyl-morpholine, N-ethylpiperidine, glucosamine, glucosamine, Histidine, breathe out amine (hydrabamine), isopropylamine, Methionin, methyl glucoside amine, morpholine, piperazine, piperidines, PROCAINE HCL, PHARMA GRADE, purine, Theobromine, triethylamine, Trimethylamine 99, tripropyl amine, three (hydroxymethyl) aminomethane (TRIS, tromethane) etc.Salt also can be by deacidite, and for example versamid 900 forms.When compound of the present invention was alkalescence, salt can be from pharmaceutically acceptable acid preparation, and described acid comprises mineral acid and organic acid.Such acid comprises acetic acid, Phenylsulfonic acid, phenylformic acid, camphorsulfonic acid, citric acid, ethyl sulfonic acid, ethionic acid, fumaric acid, glyconic acid, L-glutamic acid, Hydrogen bromide, hydrochloric acid, isethionic acid, lactic acid, toxilic acid, oxysuccinic acid, amygdalic acid, methylsulfonic acid, glactaric acid, pamoic acid, pantothenic acid, phosphoric acid, propionic acid, succsinic acid, sulfuric acid, tartrate, tosic acid etc.
Formula (I) compound can crystal or the amorphous form preparation, and if may optionally be hydrate or solvate for crystal.The present invention comprises hydrate or the solvate of chemical dose in its scope, and contains the water of different amounts and/or the compound of solvent.
In yet another aspect, the invention provides the method for preparation formula (I) compound.It will be appreciated by those skilled in the art that scheme 1-8 is by using suitable intermediate also to be applicable to the preparation of formula (I) compound, wherein R 6Be (a) (rather than described R 5Be (a)).The compound of formula (IV) is known in the literature, and R wherein 6Be (a) or R 5For the isomer of (a) is purchased.
Can be for the preparation of the square case 1 of a route of formula (IA) compound.
Figure G200880012733XD00131
Scheme 1
Formula (III) compound can be converted into formula (IV) compound by processing with hydrazine hydrate, and perhaps formula (IV) compound is commercially available maybe can prepare by described in document.
Formula (IV) compound can by under high temperature (as 50 ℃), be processed with hydroxylamine hydrochloride and suitable alkali (as sodium bicarbonate) in solvent (as methyl alcohol or ethanol) and be converted into formula (V) compound.
Formula (V) compound can under EDAC and HOBt existence, be processed with the carboxylic acid of formula (VI) in the solvent (as DMF) that is fit to and be converted into formula (VII) compound.This reaction is carried out under high temperature (as 50-80 ℃) usually.Usually, acid (VI), EDAC and HOBt in stirring at room for some time, are then added formula (V) compound.The acid of formula (VI) is maybe can preparing by following many methods of being purchased.
Formula (VII) compound can by under exist at alkali (as cesium carbonate), be processed with alkylating agent (VIII) in solvent (as DMF or DMPU) and be converted into formula (IX) compound.Described reaction can be at high temperature, as 80 ℃ or carry out in microwave reactor as at the temperature of 140 ℃.Alkylating agent (VIII) normally is purchased, but or Application standard method preparation.
Formula (IX) compound can be converted into formula (IA) compound by processing with alkali (as aqueous sodium hydroxide solution) in alcoholic solvent (as ethanol or methyl alcohol).Can add cosolvent, dissolve with help as THF.This hydrolysis reaction can carry out under room temperature or high temperature (as 50-80 ℃).Perhaps, this conversion can use solid sodium hydroxide in ethanol in microwave reactor, carry out under as 100 ℃ in temperature.Perhaps, described reaction can be in microwave reactor, uses dimethyl amine in ethanol and adds entry, carries out under as 160 ℃ in temperature.
(VII) and (VIII) reaction that generates (IX) also can produce formula (X) compound (scheme 2) of isomery.Formula (VII) compound is processed production (IX) and (X) mixture of compound with formula (VIII) alkylating agent and alkali (as cesium carbonate) in solvent (as DMF) as mentioned above, and they separate by chromatography usually.
Figure G200880012733XD00141
Scheme 2
Formula (X) compound can be converted into formula (IB) compound by the hydrolysing step described in scheme 3.
Figure G200880012733XD00142
Scheme 3
Perhaps, formula (IA) and (IB) compound can pass through the route preparation described in scheme 4,5 and 6.Formula (XI) and (XII) compound can be from formula (IV) compound by preparing with formula (VIII) alkylation reactions solvent (as DMF) under alkali (as cesium carbonate) exists.Usually, described reaction is carried out under high temperature (as 80 ℃).In some cases, compound (XI) separates by chromatography with (XII), and both mixture is used for producing subsequently the reaction of (IX) and mixture (X) in other cases, (IX) usually can separate by chromatography with the mixture of (X).
Figure G200880012733XD00151
Scheme 4
Formula (XI) compound can be converted into formula (IX) compound as described in scheme 5.
Figure G200880012733XD00152
Scheme 5
Formula (XI) compound can by in solvent (as ethanol), be converted into formula (XIII) compound lower the processing with hydroxylamine hydrochloride and alkali (as sodium bicarbonate) of high temperature (as 50 ℃).
Formula (XIII) compound can by under EDAC and HOBt existence, be processed with formula (VI) carboxylic acid in the solvent (as DMF) that is fit to and be converted into formula (IX) compound.Described reaction is carried out under high temperature (as 50-120 ℃) usually.Sometimes, acid (VI), EDAC and HOBt in stirring at room for some time, are then added formula (XII) compound, perhaps all reagent can be mixed together and heat.Formula (IX) compound can be converted into formula (IA) compound as mentioned above.
Formula (XII) compound can be converted into formula (X) compound described in scheme 6.
Scheme 6
Formula (XII) compound can by in solvent (as ethanol), be converted into formula (XIV) compound lower the processing with hydroxylamine hydrochloride and alkali (as sodium bicarbonate) of high temperature (as 50 ℃).
Formula (XIV) compound can under the existence of EDAC and HOBt, be processed with formula (VI) carboxylic acid in the solvent (as DMF) that is fit to and be converted into formula (X) compound.Described reaction is carried out under high temperature (as 80-120 ℃) usually.Acid (VI), EDAC and HOBt can at room temperature stir for some time, then add formula (IX) compound, and perhaps all reagent can be mixed together and heat.Formula (X) compound can be converted into formula (IB) compound as mentioned above.
Formula (XI) compound, wherein Z is-CH 2-CH 2-(shown in (XVIII) compound) also can be by the preparation of the route described in scheme 7.
Figure DEST_PATH_GSB00000196974300011
Scheme 7
Formula (XV) compound can be by in solvent (as chloroform), process with diacetyl oxide at the temperature of 5 ℃-15 ℃, then be converted into formula (XVI) compound with alkali (as potassium acetate) and Isopentyl nitrite processing at the temperature of room temperature to 80 ℃.
Formula (XVI) compound can by under exist at alkali (as DBU) and in solvent (as acetonitrile), be processed with the 2-ethyl propenoate under reflux temperature and be converted into formula (XVII) compound, and avoid forming isomer (XIX):
Formula (XVII) compound can pass through at catalyzer (as Pd 2(dba) 3) and under part (as DPPF) exists, in solvent (as Isosorbide-5-Nitrae-dioxs), be converted into formula (XVIII) compound with the processing of dicyano zinc under reflux temperature.
Perhaps, formula (XVI) compound can pass through at catalyzer (as Pd (PPh 3) 4) under existence, in solvent (as NMP), lower to dicyano zinc [Zn (CN) in temperature (as 80 ℃) 2] process and to be converted into formula (IV) compound.Then formula (IV) compound can use with the similarity method that formula (XV) compound is converted into the use of formula (XVII) compound and be converted into formula (XVIII) compound.
Formula (XIII) compound can be converted into formula (IX) compound by route described according to scheme 8:
Figure G200880012733XD00181
Scheme 8
Formula (VI) compound can by under existing at DMF, in solvent (as methylene dichloride), at room temperature be processed with oxalyl chloride and be converted into formula (XIX) compound.
Formula (XIII) compound can by under existing at alkali (as triethylamine), in solvent (as methylene dichloride), in 0 ℃, be converted into formula (XXI) compound with formula (XX) compound treatment.
Formula (XXI) compound can be formula (IX) compound at 100 ℃ of thermal conversions under existing at solvent (as acetonitrile).
" or formula (XIII) compound can be by in solvent (as acetonitrile), is converted into formula (IX) compound with formula (XX) compound and triethylamine processing at the temperature of room temperature to 110 ℃ ".
Therefore, formula (I) compound and their pharmacy acceptable salts are used for the treatment of by the receptor-mediated illness of S1P1 or disease.Especially, formula (I) compound is used for the treatment of multiple sclerosis, autoimmune disorder, chronic inflammatory illness, asthma, inflammatory neuropathy, sacroiliitis, transplanting, Crohn disease, ulcerative colitis, lupus erythematosus, psoriasis, ischemia reperfusion injury, noumenal tumour and metastases, with blood vessel, relevant disease, vascular disease, antalgesic, acute disease viral disease, inflammatory bowel, Regular Insulin and non-insulin-dependent diabetes mellitus (NIDDM) (hereinafter referred to " disease of the present invention ") occurs with their pharmacy acceptable salts.
Be to be understood that used herein " treatment " comprise prevention and the alleviating of definite symptom.
Therefore, the present invention also provides formula (I) compound or its pharmacy acceptable salt, and it is as particularly treating the illness that mediated by the S1P1 acceptor or the therapeutant of disease.Particularly, the invention provides formula (I) compound or its pharmacy acceptable salt, it is as the therapeutant for the treatment of multiple sclerosis, autoimmune disorder, chronic inflammatory disease, asthma, inflammatory neuropathy, sacroiliitis, transplanting, Crohn disease, ulcerative colitis, lupus erythematosus, psoriasis, ischemia reperfusion injury, noumenal tumour and metastases, disease, vascular disease, antalgesic, acute disease viral disease, inflammatory bowel, Regular Insulin and the non-insulin-dependent diabetes mellitus (NIDDM) relevant with the blood vessel generation.The illness that the present invention also provides the treatment Mammals to comprise to be mediated by the S1P1 acceptor in the people or the method for disease, described method comprises formula (I) compound or its pharmacy acceptable salt that delivers medicine to safe and effective amount on patient treatment.
On the other hand, the invention provides formula (I) compound or its pharmacy acceptable salt purposes in the medicine of the illness that is mediated by the S1P1 acceptor for the preparation for the treatment of or disease.
For use formula (I) compound in treatment, the pharmacy practice according to standard is mixed with pharmaceutical composition with them usually.The present invention also provides pharmaceutical composition, and it comprises formula (I) compound or its pharmacy acceptable salt, and pharmaceutically acceptable carrier or vehicle.
On the other hand, the invention provides the method for pharmaceutical compositions, described method comprises formula (I) compound or its pharmacy acceptable salt and pharmaceutically acceptable carrier or mixed with excipients.
Pharmaceutical composition of the present invention can be suitably by being mixed with at ambient temperature and under barometric point, be generally suitable for oral, parenteral or rectal administration, and can be with tablet, capsule, oral liquid, pulvis, granula, lozenge, restructural pulvis (reconstitutable powders) but, injectable or the solution of infusion or the form of suspension or suppository exist.The composition of common preferred oral administration.
The compounds of this invention can be by the GTP γ S test determination of carrying out on people clone's acceptor as described herein for usefulness and the effect of S1P1 acceptor, or by yeast in conjunction with test determination, use function test as herein described to prove that also formula (I) compound of description has agonist activity for the S1P1 acceptor.
Tablet and Capsula for oral administration can be unit dosage, and can contain conventional excipients, as tackiness agent (as pregelatinized W-Gum, polyvinylpyrrolidone or HYDROXY PROPYL METHYLCELLULOSE); Weighting agent (as lactose, Microcrystalline Cellulose or secondary calcium phosphate); Compressing tablet lubricant (as Magnesium Stearate, talcum or silicon-dioxide); Disintegrating agent (as yam starch or primojel); With acceptable wetting agent (as Sodium Lauryl Sulphate BP/USP).Tablet can carry out dressing by the method for knowing in common medicinal practice.
Oral liquid can be for example moisture or contain the form of oil suspension, solution, emulsion, syrup or elixir, perhaps can exist with the form of dryed product, this dryed product is water or other suitable carrier (vehicle) reconstruct (reconstitution) before use.described liquid preparation can contain conventional additives for example suspending agent (as sorbitol syrups, derivatived cellulose or hydrogenation edible-fat), emulsifying agent (as Yelkin TTS or Sudan Gum-arabic), (it can comprise edible oil such as Prunus amygdalus oil to nonaqueous carrier, grease, ethanol or fractionated vegetable oil (fractionated vegetable oils)), sanitas (as methyl p-hydroxybenzoate or propyl ester or Sorbic Acid), and if necessary, said preparation can also comprise conventional seasonings (flavouring) or tinting material, resiliency salt and sweeting agent.Oral Preparation can suitably be mixed with the controlled release release dosage form of active compound.
For administered parenterally, use compound of the present invention or its pharmacy acceptable salt and sterile carrier to be mixed with the liquid unit doses form.Injection preparation can use compound of the present invention or its pharmacy acceptable salt and sterile carrier, randomly adds the sanitas preparation, for example is contained in ampoule or exists with multiple doses with unit dosage.Composition can be taked the form of suspension, solution or emulsion in oiliness or aqueous carrier, and can contain reagent preparation, as suspending agent, stablizer and/or dispersion agent.On the other hand, activeconstituents can be powder type, uses before use suitable carrier, builds (costitution) as aseptic apirogen water.According to carrier used and concentration, described compound can be suspended or is dissolved in described carrier.In the process of obtain solution, described compound dissolution can be used for injection and filtration sterilization, be filled into subsequently in suitable bottle (vial) or ampoule, then sealing.Advantageously, for example local anesthetic, sanitas and buffer reagent are dissolved in carrier with auxiliary agent.In order to improve stability, after can being filled into composition in bottle, with it freezing and under vacuum except anhydrating.Except being suspended in described compound in carrier rather than being dissolved in carrier, and can not be by outside filtration sterilization, with substantially the same method preparation parenteral suspension.Before in compound is suspended in sterile carrier, can sterilize to described compound by being exposed in oxyethane.Advantageously, tensio-active agent or wetting agent are joined in described composition, in order to promote the even distribution of compound.
Lotion can be prepared with water-based or oleaginous base, and lotion generally also comprises one or more emulsifying agents, stablizer, dispersion agent, suspending agent, thickening material or tinting material.Drops can use or non-aqueous substrate preparation, also comprises one or more dispersion agents, stablizer, solubilizing agent or suspending agent.They also can contain sanitas.
Compound of the present invention can also be made the form of rectal compositions, as suppository or enema,retention, for example contains conventional suppository bases such as theobroma oil or other glyceryl ester.
Compound of the present invention also can be formulated into prolonged action preparation (depot preparation).These prolonged action preparations can be by implanting (implantation) mode (for example subcutaneous or intramuscular) or passing through the muscle injection mode administration.Therefore, for example compound of the present invention can with suitable polymkeric substance or hydrophobic material (for example with the emulsion in can accepting oil) or ion exchange resin, or with the microsolubility derivative for example slightly soluble salt make together preparation.
For intranasal administration, compound of the present invention or its pharmacy acceptable salt can be formulated into the solution by suitable metering or the administration of single dose device, perhaps are mixed with powdered mixture with the carrier that is fit to, and utilize the drug delivery systems administration that is fit to.Thereby that formula (I) compound can be formulated into is oral, suck, parenteral, part (comprising eye and nose), slowly-releasing (depot) or rectal administration formulation or be formulated into the formulation that is suitable for by sucking or be blown into (by mouth or nose) administration.
Formula (I) compound or its pharmacy acceptable salt can be formulated into ointment, creme, gelifying agent, lotion, vaginal suppository, aerosol or the drops (for example eye, ear or nose drops) for topical.Ointment and creme for example can use or oleaginous base prepared, wherein add suitable thickening material and/or jelling agent.Dosing eyes with ointment can use through the sterilization component, prepared by sterile manner.
According to medication, described composition can contain 0.1%～99 % by weight, preferably contains the active substance of 10～60 % by weight.The dosage that is used for the treatment of the compound of aforementioned diseases can change with the severity of disease, patient's body weight and other similar factors in a conventional way.But as general guidance, suitable unitary dose can be 0.05～1000mg, 1.0～500mg or 1.0～200mg; And described unitary dose can every day multiple dosing, for example every day two or three administrations.
Formula (I) compound or its pharmacy acceptable salt can combination preparation (combinationpreparations) form use.For example, the compounds of this invention can use with the compound combination of Ciclosporin A or other treatment activity.
The present invention also comprises isotope-labeled compound, its with formula (I) and below described in those Compound Phases with, but in fact one or more atom is had and is different from occurring in nature usually the nucleidic mass of existence or the nucleidic mass of total mass number or the atom of total mass number are replaced.Can be incorporated into the isotope that isotopic example in compound of the present invention comprises hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, iodine and chlorine, as 3H, 11C, 14C, 18F, 123I and 125I.
Contain the pharmacy acceptable salt of other isotopic formula (I) compound of above-mentioned isotropic substance and/or other atom and described compound within the scope of the invention.The isotope-labeled compound of the present invention (for example mix radio isotope as 3H、 14The compound of C) can be used for medicine and/or substrate tissue distribution assays.Tritium generation, namely 3H, and carbon-14, that is, 14C, isotropic substance is particularly preferred, because they are easy to preparation and detect. 11C and 18The F isotope especially can be used for PET (positron emission tomography), and 125The I isotope is used in particular for SPECT (single photon emission computerized tomography), and they are all for the brain imaging.In addition, with heavier isotropic substance (as deuterium, namely 2H) replace some treatment advantage can be provided, obtain better metabolic stability, the Half-life in vivo that for example increases or the dosage demand of reduction, and therefore in some cases heavier isotropic substance be preferred.Isotope-labeled formula of the present invention (I) compound and following compound generally can replace nonisotopically labelled reagent to be prepared by being implemented in following proposal and/or disclosed method in an embodiment, utilizing the isotope-labeled reagent that is easy to get.
All publications of quoting in this manual including, but not limited to patent and patent application, are incorporated herein by reference them at this, just as each independent publication is fully quoted as a reference at this particularly and individually with setting forth.
The following describes with embodiment and for example understand the preparation of the compounds of this invention.
The following explanation that provides the hardware and software of analytical procedure is only for illustrative purposes.
For condition, the hardware and software of analyzing the LCMS system
Hardware
Agilent 1100 gradient pumps
Agilent 1100 automatic samplers
Agilent 1100DAD detector
Agilent 1100 degasifiers
Agilent 1100 thermostat containers
Agilent 1100 controllers
Waters Acquity Binary solvent controller
Waters Acquity sample controller
Waters Acquity PDA
Waters ZQ mass spectrograph
Sedere Sedex 55, Sedere Sedex 85, Sedere Sedex 75 or Polymer LabsPL-ELS-2100
Software
Waters MassLynx edition 4 .0 SP2 or 4.1
For 5 minutes clock methods
Pillar
Pillar used is Waters Atlantis, and its size is 4.6mmx50mm.The stationary phase particle diameter is 3 μ m.
Solvent
A: water-containing solvent=water+0.05% formic acid
B: organic solvent=acetonitrile+0.05% formic acid
Method
Universal method used has the working time of 5 minutes.
Time/minute %B
0 3
0.1 3
4 97
4.8 97
4.9 3
5.0 3
Flow velocity
The flow velocity of aforesaid method is 3ml/ minute.
For 2 minutes clock methods
Software
Waters MassLynx edition 4 .1
Pillar
Pillar used is Waters Acquity BEH UPLC C18, and its size is 2.1mmx50mm.The stationary phase particle diameter is 1.7 μ m.
Solvent
A: water-containing solvent=water+0.05% formic acid
B: organic solvent=acetonitrile+0.05% formic acid
Weak wash-out (Weak Wash)=1: 1 methyl alcohol: water
Strong wash-out (Strong Wash)=water
Method
All general method has the working time of 2 minutes.
Time/minute ％B
0 3
0.1 3
1.5 97
1.9 97
2.0 3
The flow velocity of aforesaid method is 1ml/ minute
The volume injected of universal method is 0.5 μ l
Column temperature is 40 ℃
The UV sensing range is 220-330nm
Open Access Mass Directed Auto Prep System (the automatic preparative liquid chromatography system that Open Access mass spectrum is controlled) (MDAP)
Hardware
Open Access Mass Directed Prep instrument comprises following equipment:
1 Waters 600 gradient pumps
1 Waters 2767 injection/collectors
1 Waters reagent controller
1 MicroMass ZQ mass spectrograph
1 Gilson Aspec-liquid waste collector
The 1 rear fraction UV detector of Gilson 115
1 computer system.
Software
MicroMass MassLynx v4.0
Pillar
Pillar used is generally Supelco LCABZ++ pillar, and its size is internal diameter 20mm, long 100mm.The stationary phase particle diameter is 5 μ m.
Solvent
A: water-containing solvent=water+0.1% formic acid
B:. organic solvent=MeCN: water 95: 5+0.05% formic acid
Replenish (Make up) solvent=MeOH: water 80: 20+50mMol ammonium acetate
Pin rinse solvent=MeOH: water: DMSO 80: 10: 10
Method
Can use a kind of in 5 kinds of methods according to the analysis retention time of concern compound.
Be 15-minute methodical working time, comprises 10-minute gradient elution, and then 5-minute pillar rinses and the reequilibrate step.
MDP 1.5-2.2＝0-30％B
MDP 2.0-2.8＝5-30％B
MDP 2.5-3.0＝15-55％B
MDP 2.8-4.0＝30-80％B
MDP 3.8-5.5＝50-90％B
Flow velocity
The flow velocity of all aforesaid methods is 20ml/ minute.
Another kind of equipment:
Hardware:
Waters 2,525 two gradient modules (Binary Gradient Module)
Waters 515 make-up pumps (makeup pump)
Waters pump control module (Pump Control Module)
(Inject Collect) collected in Waters 2767 injections
Waters pillar Flow Control controller (Column Fluidics Manager)
Waters 2996 photodiode array detectors (Photodiode Array Detector)
Waters ZQ mass spectrograph
Gilson 202 fraction collectors
Gilson Aspec liquid waste collector
Software
Waters MassLynx edition 4 SP2
Pillar
Pillar used is Waters Atlantis, and its size is 19mmx100mm (in a small amount) and 30mmx100mm (in a large number).The stationary phase particle diameter is 5mm.
Solvent
A: water-containing solvent=water+0.1% formic acid
B: organic solvent=acetonitrile+0.1% formic acid
Supplementing solvent=methyl alcohol: water 80: 20
Pin rinse solvent=methyl alcohol
Method
There are 5 kinds of methods to use according to the analysis retention time of concern compound.They have the working time of 13.5-minute, comprise 10-minute gradient elution, and then 3.5 minutes pillars rinse and the reequilibrate step.
In a large number/a small amount of 1.0-1.5=5-30%B
In a large number/a small amount of 1.5-2.2=15-55%B
In a large number/a small amount of 2.2-2.9=30-85%B
In a large number/a small amount of 2.9-3.6=50-99%B
In a large number/a small amount of 3.6-5.0=80-99%B (6 minutes, then rinsed and reequilibrate in 7.5 minutes)
Flow velocity
The flow velocity of all above-mentioned steps is 20mls/ minute (in a small amount) or 40mls/ minute (in a large number).
Shallow gradient (Shallow gradients)
A large amount of 1.5-2.3 minute=13-29%B
A large amount of 1.9-2.3 minute=25-41%B
A large amount of 2.3-2.6 minute=37-53%B
A large amount of 2.6-3.1 minute=49-65%B
A large amount of 3.1-3.6 minute=61-77%B
For NMR condition used
Hardware
Bruker 400MHz Ultrashield
Bruker B-ACS60 automatic sampler
Bruker Advance 400 supervisory control desks
Bruker DPX250
Bruker AVANCE 500
Bruker DRX600
Software
User interface-NMR Kiosk
Controlling software-XWin NMR version 3 .0
Chromatography
Except as otherwise noted, all chromatographys use silicagel column to carry out.
Abbreviation:
The g-gram
The mg-milligram
The ml-milliliter
μ l-microlitre
CHCl 3-chloroform
The MeCN-acetonitrile
MeOH-methyl alcohol
EtOH-ethanol
The EtOAc-ethyl acetate
The DBA-dibenzalacetone
The DCM-methylene dichloride
DMF-N, dinethylformamide
DMPU-1,3-dimethyl-3,4,5,6-tetrahydrochysene-2 (1H)-pyrimidone
The DMSO-dimethyl sulfoxide (DMSO)
DPPF-1,1 '-two (diphenylphosphino) ferrocene
EDC-N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride
EDAC-N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride
EDCl.HCl-N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride
HATU-2-(1H-7-azepine benzo triazol-1-yl)-1,1,3,3-tetramethyl-urea hexafluorophosphate first ammonium
The IPA-Virahol
The NMP-N-N-methyl-2-2-pyrrolidone N-
PyBOP-benzotriazole-1-base-oxygen base three (pyrrolidyl) Phosphonium hexafluorophosphate
The THF-tetrahydrofuran (THF)
The dba-dibenzalacetone
The RT-room temperature
℃-degree centigrade
The M-volumetric molar concentration
The H-proton
S-is unimodal
The d-doublet
The t-triplet
The q-quartet
The LCMS-Liquid Chromatography-Mass Spectrometry
The LC/MS-Liquid Chromatography-Mass Spectrometry
The NMR-nucleus magnetic resonance
The MS-mass spectrum
The ES-electron spray(ES)
MH +-mass ion (mass ion)+H +
The automatic preparative liquid chromatography (Mass Directed AutomatedPreparative liquid chromatography) that the MDAP-mass spectrum is controlled
The SCX-strong cation exchange
General chemistry
The intermediate of Preparation Example is not necessarily with described particular batch preparation.
The description of D1
1H-indazole-5-formonitrile HCN (D1)
Figure G200880012733XD00291
2-fluoro-5-cyanobenzaldehyde (1.1g) is dissolved in hydrazine hydrate (19ml), and in stirring at room 18 hours.Then retort solution, and resistates is by silica gel flash column chromatography (using the DCM wash-out) purifying.Obtain title compound (300mg).
The description of D1 (another kind of method)
1H-indazole-5-formonitrile HCN (D1)
Figure G200880012733XD00292
5-cyano group-3-fluorobenzaldehyde (4g, 26.8mmol) is dissolved in hydrazine hydrate, and in stirring at room 18 hours.Solution is poured on ice, and the separation of gained solid by filtration, then obtain title compound (1.27g) in 60 ℃ of dryings under vacuum, be pink solid.Separate other material from water layer.MS (ES): C 8H 5N 3Calculated value 143; Measured value 144 (MH +).
1H-indazole-5-formonitrile HCN also can be according to former description preparation (Halley and Sava 1997, Synth.Commun.27 (7): 1199-1207).
The description of D2
N-hydroxyl-1H-indazole-5-carbonamidine (D2)
Figure G200880012733XD00293
With 5-cyano group indazole (D1) (500mg), azanol .HCl (485mg) and NaHCO 3(1.47g) dissolve/be suspended in MeOH (18ml), be heated to 50 ℃, and stirred 19 hours, then spend the night.Then filter reaction mixture, with MeOH washing, and evaporation obtains title compound (584mg), is white solid.MS (ES): C 8H 8N 4O calculated value 176; Measured value 177 (MH +).
The description of D3
3-chloro-4-[(1-methylethyl) oxygen base] phenylformic acid (D3)
4-hydroxyl-3-chloro benzoic ether (13.4g) is dissolved in DMF (150ml), uses K 2CO 3(19.9g) process, then use isopropyl bromide (13.5ml) to process, then with gained mixture heating up to 70 ℃, and stir and spend the night.Then reaction mixture is cooled to room temperature, is evaporated to driedly, then be dissolved in EtOH, filter and again evaporation obtain intermediate ester (22.24g), be white solid.This compound is 3: 1 mixtures of ethyl ester and methyl ester, and uses crude product in next step reaction.
Thick intermediate (22.24g) is dissolved in MeOH (75ml), processes with the NaOH aqueous solution (75ml) of 2M, be heated to 60 ℃, and stirred 2 hours.Then reaction mixture is cooled to room temperature, evaporation MeOH, and with the HCl aqueous solution (30ml) acidifying of the remaining aqueous solution with 5M.Leach precipitation and drying and obtain title compound (15.1g), be white solid.δ H (CDCl 3, 400MHz): 1.42 (6H, d), 4.70 (1H, septets), 6.97 (1H, d), 7.97 (1H, d), 8.12 (1H, s) .MS (ES): C 10H 11ClO 3Calculated value 214; Measured value 213 (M-H +).
3-chloro-4-[(1-methylethyl) oxygen base] phenylformic acid can prepare according to described in WO2006047195.
The description of D4
5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole (D4)
Figure G200880012733XD00302
With 3-chloro-4-[(1-methylethyl) the oxygen base] phenylformic acid (D3) (712mg), EDCI.HCl (699mg) and HOBT (493mg) are dissolved in DMF (16.5ml), and in stirring at room 10 minutes.Add N-hydroxyl-1H-indazole-5-carbonamidine (D2) (584mg), and with mixture heating up to 80 ℃, kept 12 hours.Then reaction mixture is cooled to room temperature, and standing weekend excessively, be evaporated to dry doubling and be dissolved in again H 2In O.This solution is with EtOAc (x3) extraction, and the extraction liquid salt water washing that merges, and evaporation is also ground 2 times with DCM, and then supersound process 4 minutes filters, drying and repeat 1 time and obtain title compound (407mg), is pink solid.δ H (d 6-DMSO, 400MHz): 1.37 (6H, d), 4.89 (1H, septet), 7.46 (1H, d), (7.73 1H, d), 8.04 (1H, d), 8.13 (1H, d), (8.20 1H, s), 8.28 (1H, s), 8.58 (1H, s) .MS (ES): C 18H 15ClN 4O 2Calculated value 354; Measured value 355 (M+H).
The description of D5
3-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] ethyl propionate (D5)
Figure G200880012733XD00311
With 5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole (D4) (200mg) is dissolved in DMPU (2.8ml), and uses Cs 2CO 3(367mg) process, then use 3-ethyl bromide (145 μ l) to process.The gained mixture is heated to 140 ℃ in microwave reactor, kept 1.5 hours.Then reaction mixture is cooled to room temperature, is evaporated to driedly, and be dissolved in H 2In O.This solution is with EtOAc (x3) extraction, and the extraction liquid salt water washing that merges, and evaporates and obtain title compound (240mg), is brown oil.MS (ES): C 23H 23 35ClN 4O 4Calculated value 454; Measured value 455 (MH +).
The description of D5 (another kind of method)
3-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] ethyl propionate (D5)
Figure G200880012733XD00312
With the 3-{5-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl } ethyl propionate (D52) (2.04g, 7.38mmol), 3-chloro-4-[(1-methylethyl) oxygen base] phenylformic acid (D3) (1.743g, 8.12mmol), EDC (2.123g, 11.08mmol) and HOBT (1.357g, 8.86mmol) at N, the solution stirring of dinethylformamide (DMF) in (25ml) spent the night, then in 4 hours in 120 ℃ of heating.Mixture is dissolved in EtOAc, then water and NaHCO 3Washing, and by purified by flash chromatography is with 10-90% 10-30%EtOAc/ isohexane wash-out then.Evaporating solvent obtains the white solid of (1.95g).MS (ES): C 23H 23 35ClN 4O 4Calculated value 454; Measured value (MH +) 455.
The description of D6
3-chloro-4-(oxyethyl group) methyl benzoate (D6)
Figure G200880012733XD00321
4-hydroxyl-3-chloro benzoic ether (10g, 53.6mmol) and salt of wormwood (14.8g, 107.2mmol) are suspended in DMF (110ml), then add iodoethane (8.56ml, 107.2mmol).Reaction mixture is heated to 70 ℃ to spend the night.Filter reaction mixture, and resistates washs with ether.Evaporate in a vacuum organic solution, then be dissolved in EtOAc, and with the NaOH aqueous solution and water washing, dry and be evaporated in a vacuum the dried title compound (11.24g) that obtains, be yellow oil.MS (ES): C 10H 11 35ClO 3Calculated value 214; Measured value (MH +) 215.
The description of D7
3-chloro-4-(oxyethyl group) phenylformic acid (D7)
Figure G200880012733XD00322
Add the NaOH aqueous solution (40ml) of 2M in the suspension of 3-chloro-4-(oxyethyl group) methyl benzoate (D6) (11.24g, 52.4mmol) in methyl alcohol (40ml) that stirs, and the gained mixture was heated 2 hours in 60 ℃.Reaction mixture is allocated in EtOAc and dilute hydrochloric acid.Separate organic layer, dry also the evaporation in a vacuum obtains crude product.This material grinds with ether and obtains title compound, is white solid (8.21g).MS (ES): C 9H 9 35ClO 3Calculated value 200; Measured value (MH +) 201.
The description of D8
3-ethyl-4-(piperidino) benzonitrile (D8)
With 4-amino-3-ethyl benzonitrile (3.0g, 20.5mmol), 1, pentamethylene bromide (11.1mL, 82.1mmol), salt of wormwood (5.67g, 41.0mmol) and water (39.6mL) all deciles in 10 microwave tubes, and with each 160 ℃ of heating 1 hour.Merge all reaction mixtures, and extract 2 times with ethyl acetate (40mL), and with the organic moiety dry (phase splitter) that merges, and concentrated in a vacuum.Add methylene dichloride, filtering mixt, then filtrate is by the silica gel chromatography purifying, and the hexane solution wash-out with the 2-5% ethyl acetate obtains title compound, is colorless oil (823mg, 3.85mmol).The analysis showed that this compound contains a small amount of dibromo pentane impurity.MS (ES): C 14H 18N 2Calculated value 214; Measured value 215 (MH +).
The description of D9
3-ethyl-4-(piperidino) phenylformic acid (D9)
Figure G200880012733XD00332
With 3-ethyl-4-(piperidino) benzonitrile (the D8) (817mg in ethanol (35mL) and water (8mL), 3.82mmol) and potassium hydroxide (2.14g, 38.2mmol) be heated to 90 ℃ (device temperature (blocktemperature)), kept 9 hours.Add other potassium hydroxide (2.14g, 38.2mmol) and water (8mL), and reaction reheats 18 hours.Make reaction cooling, and neutralize with the HCl aqueous solution.White solid passes through filtering separation, and attempts by the SCX column purification, but failure.The product of combining solid and SCX adds methyl alcohol, then mixture acetic acid acidifying.Filtering mixt obtains filtrate, then filtrate is added in the SCX post, uses methanol wash, and with the methanol solution wash-out of 2M ammonia.Obtain title compound for experimental scale, be white solid (96mg, 0.41mmol), and obtain colorless oil (563mg, 2.41mmol) for residual substance.MS (ES): C 14H 19NO 2Calculated value 233; Measured value 234 (MH +).
The description of D10
3-ethyl-4-iodine benzonitrile (D10)
Figure G200880012733XD00341
In 0 ℃, drip concentrated hydrochloric acid (7.80mL, 257mmol) in the 4-amino that the stirs-solution of 3-ethyl benzonitrile (2.50g, 17.1mmol) in water (14mL), then drip the solution of Sodium Nitrite (1.24g, 18.0mmol) in water (3.43mL).The gained mixture was stirred 15 minutes, then join in the solution of potassiumiodide (2.98g, 18.0mmol) in water (6.0mL) through 15 minutes in 0 ℃.Mixture was in stirring at room 2 hours.Mixture extracts with ethyl acetate (3x100mL), and the organic moiety that merges salt solution (100mL) washing, and dry (phase splitter) is also concentrated in a vacuum obtains title compound, is brown solid (4.21g, 16.4mmol).δ H (methyl alcohol-d 4, 400MHz): 8.02 (1H, d), 7.61 (1H, d), 7.24 (1H, dd), 2.80 (2H, q), 1.21 (3H, t).
The description of D11
4-(1-tetrahydrobenzene-1-yl)-3-ethyl benzonitrile (D11)
Figure G200880012733XD00342
With 3-ethyl-4-iodine benzonitrile (D10) (1.23g, 4.80mmol), 1-tetrahydrobenzene-1-ylboronic acid (907mg, 7.20mmol), sodium methylate (778mg, 14.4mmol) and chlorination two (triphenylphosphines) close the mixture of palladium (II) (337mg, 0.48mmol) in anhydrous methanol (12mL) in 80 ℃ of heating 10 minutes in microwave.Reaction mixture is allocated in ethyl acetate (40mL) and water (40mL), the then further water of organic layer (40mL) washing, dry (phase splitter) is also concentrated in a vacuum.Thick material through 30 minutes hexane solution wash-outs with 0-5%EtOAc, obtains title compound by the silica gel chromatography purifying, is yellow oil (824mg, 3.91mmol).δ H (methyl alcohol-d 4, 400MHz) 7.56 (1H, d), 7.46 (1H, dd), 7.19 (1H, d), 5.61-5.56 (1H, m), 2.68 (2H, quart), 2.23-2.16 (4H, m), 1.85-1.68 (4H, m), 1.20 (3H, t).
The description of D12
4-(1-tetrahydrobenzene-1-yl)-3-ethyl benzoate (D12)
Figure G200880012733XD00351
With 4-(1-tetrahydrobenzene-1-yl)-3-ethyl benzonitrile (D11) (824mg, 3.91mmol) and potassium hydroxide (2.19g, 39.1mmol) in ethanol (36mL) and water (8mL) in 90 ℃ (device temperature) heating 20 hours.Reaction mixture is concentrated in a vacuum, and resistates is allocated in ethyl acetate (120mL) and aqueous hydrochloric acid (2M, 50mL), then organic phase is with other hydrochloric acid (2M, 50mL) washing, dry (phase splitter), and vacuum concentration obtains title compound, be yellow oil (808mg, 3.51mmol).LCMS (ES): C 15H 18O 2Calculated value 230; Measured value 229 (M-H +).
The description of D13
4-cyclohexyl-3-ethyl benzoate (D13)
Figure G200880012733XD00361
4-(1-tetrahydrobenzene-1-yl)-3-ethyl benzoate (D12) (803mg, 3.49mmol) is dissolved in methyl alcohol (70mL), and uses palladium/carbon post hydrogenation on H-Cube.Product solution is concentrated in a vacuum obtain title compound, be white solid (792mg, 3.41mmol).δ H (methyl alcohol-d 4, 400MHz): 7.82-7.68 (2H, m), 7.33 (1H, d), 2.83 (1H, m), 2.73 (2H, q), 1.87 (2H, m), 1.85-1.70 (3H, m), 1.58-1.30 (5H, m), 1.22 (3H, t).
The description of D14
3-bromo-4-[(1-methylethyl) oxygen base] phenylformic acid 1-methyl ethyl ester (D14)
Figure G200880012733XD00362
With 3-bromo-4-HBA (2.00g, 9.22mmol), 2-iodopropane (1.85mL, 18.4mmol) and salt of wormwood (2.55g, the 18.4mmol) mixture heating up in DMF (175mL) refluxed 5 hours.To react cooling and filter.Filtrate is concentrated in a vacuum, and resistates is allocated in ethyl acetate (150mL) and water (150mL), alkalizes with the NaOH of 2M.With organic phase dry (phase splitter), and vacuum concentration obtains title compound, is yellow oil (2.36g, 7.84mmol).δ H (methyl alcohol-d 4, 400MHz): 8.05 (1H, d), 7.90 (1H, dd), 7.25 (1H, d), 5.10 (1H, septets), 4.81 (1H, septets), 1.32 (6H, d), 1.31 (6H, d).
The description of D15
3-bromo-4-[(1-methylethyl) oxygen base] phenylformic acid (D15)
Figure G200880012733XD00371
With 3-bromo-4-[(1-methylethyl) the oxygen base] vlil of phenylformic acid 1-methyl ethyl ester (D14) (2.36g, 7.84mmol) in ethanol (100mL) and aqueous sodium hydroxide solution (2M, 39mL) 5 hours.Reaction mixture is concentrated in a vacuum, and is allocated in ethyl acetate (125mL) and water (125mL), water layer HCl (40mL) acidifying of 2M.Water layer is used ethyl acetate (70mL) extraction again, and with the organic extract liquid dry (phase splitter) that merges, and vacuum concentration obtains title compound, is pale solid (1.83g, 7.06mmol).MS (ES): C 10H 11BrO 3Calculated value 258,260; Measured value 257,259 (M-H +).
The description of D16
4-bromo-3-chloro-benzoic acid ethyl ester (D16)
Figure G200880012733XD00372
Add sulfuric acid (5mL) in the suspension of 4-bromo-3-chloro-benzoic acid (5.00g, 21.2mmol) in ethanol (50mL), and the gained mixture heating up was refluxed 60 hours.Reaction is allocated in ethyl acetate (50mL) and water (50mL).Water layer is used ethyl acetate extraction again, and with the organic moiety dry (phase splitter) that merges, and vacuum concentration obtains title compound, is brown oil/solid (5.09g, 19.3mmol).δH(d 6-DMSO，400MHz)：8.06(1H，d)，7.96(1H，d)，7.80(1H，dd)，4.33(2H，q)，1.33(3H，t)。
The description of D17
3-chloro-4-(2-methyl-propyl) ethyl benzoate (D17)
Figure G200880012733XD00381
Under argon gas, with the solution (0.5M of bromination isobutyl-zinc in THF, 30mL, 15.0mmol) join 4-bromo-3-chloro-benzoic acid ethyl ester (D16) (2.00g, 7.60mmol) in, then add dichloride 1,1 '-two (diphenylphosphino) ferrocene-palladium (II) methylene dichloride mixtures (930mg, 1.14mmol).To react reflux 4.5 hours.Mixture is concentrated in a vacuum, and resistates is allocated in ethyl acetate (125mL) and water (125mL).Form solid, leach and discard.Organic layer water (100mL) washing, dry (phase splitter), and vacuum concentration.Crude product through 30 minutes hexane solution wash-outs with 0-5%EtOAc, obtains title compound by the silica gel chromatography purifying, is colorless oil (1.76g, 7.33mmol).δH(d 6-DMSO，400MHz)：7.91(1H，d)，7.80(1H，dd)，7.46(1H，d)，4.30(2H，q)，2.66(2H，d)，1.88-2.01(1H，m)，1.32(3H，t)，0.89(6H，d)。
The description of D18
3-chloro-4-(2-methyl-propyl) phenylformic acid (D18)
Figure G200880012733XD00382
Solution in ethanol (30mL) was in 40 ℃ of heating 3 hours with 3-chloro-4-(2-methyl-propyl) ethyl benzoate (D17) (1.76g, 7.33mmol) and aqueous sodium hydroxide solution (2M, 3.70mL, 7.4mmol).Reaction mixture is concentrated in a vacuum, and resistates is allocated in ethyl acetate (100mL) and water (100mL), HCl (4mL) acidifying of water layer use 2M.Water layer is with ethyl acetate (100mL) extraction, and with the organic extract liquid dry (phase splitter) that merges, and vacuum concentration obtains title compound, is white solid (1.35g, 6.36mmol) .MS (ES): C 11H 13 35ClO 2Calculated value 212; Measured value 211 (M-H +).
The description of D19
3-cyano group-4-{[(trifluoromethyl) alkylsulfonyl] the oxygen base } methyl benzoate (D19)
Figure G200880012733XD00391
In 0 ℃ under argon gas stream, to 3-cyano group-4-HBA methyl esters (3g, 16.93mmol) and the solution of triethylamine (3.54ml, 25.4mmol) in anhydrous methylene chloride (60ml) in slowly drip Trifluoromethanesulfonic anhydride (3.15ml, 18.63mmol).Reaction is warmed to room temperature, and stirred 1 hour.Reaction mixture washs with 10% wet chemical (2x50mL), then use the HCl aqueous solution (2M, 2x50mL) washing, then with organic phase dry (phase splitter), and remove in a vacuum solvent, and obtain title compound, be Vandyke brown oily matter, (5.165g, 16.70mmol).δH(CDCl 3，400MHz)：8.44(1H，d)，8.38(1H，dd)，7.60(1H，d)，3.99(3H，s)。
The description of D20
2-cyano group-4-diphenic acid methyl esters (D20)
Figure G200880012733XD00392
Following reaction is carried out in two batches with the amount of half: with 3-cyano group-4-{[(trifluoromethyl) alkylsulfonyl] the oxygen base } methyl benzoate (D19) (1.5g; 4.85mmol); phenyl-boron dihydroxide (1.183g; 9.70mmol); salt of wormwood (2.011g; 14.55mmol) and four (triphenylphosphines) close palladium (0) (0.561g, 0.485mmol) and be dissolved in DMF (24ml), and mixture is in 150 ℃ of heating 30 minutes in microwave.Two reaction mixtures are merged, and dilute with ethyl acetate (50mL), and mixture by diatomite filtration to remove the palladium resistates.
Filtrate is concentrated to reduce the amount of DMF in a vacuum, then resistates is allocated in saturated sodium bicarbonate aqueous solution (50mL) and ethyl acetate (50mL).Organic phase is used sodium bicarbonate (50mL) washing again, then water (50mL) washing, then dry (MgSO 4), filter, and remove in a vacuum solvent.Brown solid through 35 minutes isohexane eluant solutions with 0-25%EtOAc, obtains title compound by the silica gel chromatography purifying, is white solid (935mg, 3.94mmol).δH(d 6-DMSO，400MHz)：8.42(1H，d)，8.29(1H，dd)，7.81(1H，d)，7.65(2H，m)，7.60-7.50(3H，m)，3.92(3H，s)。
The description of D21
2-cyano group-4-diphenic acid (D21)
Figure G200880012733XD00401
Add ethanol (18ml) in 2-cyano group-4-diphenic acid methyl esters (D20) (935mg, 3.94mmol), but therefore not dissolving adds methylene dichloride (10ml).Then add 2M aqueous sodium hydroxide solution (2ml, 4.00mmol), stirring reaction 2 hours.The HCl aqueous solution (10mL) that adds methylene dichloride (20mL) and 2M in the mixture.Separate each layer, and water layer extracts with other methylene dichloride (20mL).With the organic phase dry (phase splitter) that merges, and remove in a vacuum solvent and obtain white solid, it is dissolved in methyl alcohol (30mL), and adds aqueous sodium hydroxide solution (2M, 3mL).React on stirring at room 1 hour, and then added entry (20mL).Stirring reaction is 1 hour again.Add methylene dichloride (60mL), and the jolting mixture, each layer then separated.Water is with other methylene dichloride (50mL) extraction, then with the organic phase dry (phase splitter) that merges, and removes in a vacuum solvent and obtains title compound, is white solid (849mg, 3.80mmol).MS (ES): C 14H 9NO 2Calculated value 223; Measured value 222 (M-H +).
The description of D22
4-chloro-3-(trifluoromethyl) ethyl benzoate (D22)
4-chloro-3-(trifluoromethyl) phenylformic acid (1g, 4.45mmol) is dissolved in ethanol (3ml), and adds the vitriol oil (0.15ml).Mixture in 100 ℃ of heating 5 minutes, then heated 15 minutes in 120 ℃ in microwave.Remove in a vacuum solvent, and resistates is allocated in saturated sodium bicarbonate aqueous solution (50ml) and ethyl acetate (50ml).Water layer is with other EtOAc (50ml) extraction, and merges organic phase, and is dry with phase splitter, and vacuum concentration obtains title compound (1.026g), is colorless oil.δ H (methyl alcohol-d 4, 400MHz) 1.40 (3H, t), 4.41 (2H, q), 7.76 (1H, d), 8.21 (1H, dd), 8.33 (1H, d).
The description of D22 (another kind of method)
4-chloro-3-(trifluoromethyl) ethyl benzoate (D22)
Figure G200880012733XD00412
The solution of 4-chloro-3-trifluoromethylbenzoic acid (10g, 44.5mmol) in ethanol (10ml) is distributed in two microwave tubes.Add the vitriol oil (0.75ml) (amounting to 1.5ml) in each pipe.Reaction in microwave in 120 ℃ of heating 30 minutes altogether.Merge reaction mixture and vacuum concentration.Resistates is allocated in EtOAc (100ml) and sodium bicarbonate aqueous solution (100ml), separate organic phase, with sodium bicarbonate aqueous solution (100ml) and water (2x100ml) washing, then dry (phase splitter), and remove in a vacuum solvent and obtain title compound (4.126g), be colorless oil.δ H (400MHz, methyl alcohol-d 4), consistent with the embodiment of front.
The description of D23
2-(trifluoromethyl)-4-diphenic acid (D23)
To react one and be divided into 4, each uses 1/4 reagent: to 4-bromo-3-(trifluoromethyl) benzonitrile (4g, 16.00mmol), phenyl-boron dihydroxide (3.90g, 32.0mmol) and salt of wormwood (6.63g, 48.0mmol) add four (triphenylphosphines) to close palladium (0) (1.849g, 1.600mmol) in mixture at DMF (DMF) in (64ml).Each reaction was heated 30 minutes in 150 ℃ in microwave.The reaction mixture that merges filters by Sai Lite diatomite (celite), with the ethyl acetate washing, and removes in a vacuum solvent.Resistates is allocated in ethyl acetate (100mL) and water (100mL), and organic phase is washed with sodium hydrogen carbonate solution (100mL).With the dry (MgSO of organic phase 4), filter and remove in a vacuum solvent.Brown oil is ground with methylene dichloride, and filters and to obtain faint yellow solid, 2-(trifluoromethyl)-4-dibenzoyl amine (2.47g), and it just is not further purified and uses.Add potassium hydroxide (4.23g, 75mmol) and water in the solution of 2-(trifluoromethyl)-4-dibenzoyl amine (2g, 7.54mmol) in ethanol (80ml), and with mixture heating up to 90 ℃, kept 18 hours.Reaction mixture is concentrated in a vacuum, and resistates is allocated in the HCl (100mL) of methylene dichloride (100mL) and 2M.Separate organic phase, and dry (phase splitter), and remove in a vacuum solvent and obtain crude product.Use Biotage Horizon, the reversed-phase column purifying with the aqueous solution wash-out of 5-100%MeCN, obtains the title compound (960mg) of pale solid.MS (ES): C 14H 9F 3O 2Calculated value 266; Measured value 265 (M-H +).
The description of D23 (another kind of method)
2-(trifluoromethyl)-4-diphenic acid (D23)
Batch A: with 4-chloro-3-(trifluoromethyl) ethyl benzoate (D22) (1.0g, 3.96mmol), phenyl-boron dihydroxide (724mg, 5.94mmol), acid chloride (44mg), (dicyclohexyl phosphino-) biphenyl (140.2mg) and Potassium monofluoride (689mg, the 11.9mmol) mixture in THF (8ml) in microwave in 120 ℃ of heating 40 minutes altogether.
Batch B:4-chloro-3-(trifluoromethyl) ethyl benzoate (D22) (500mg, 1.98mmol), phenyl-boron dihydroxide (290mg, 2.38mmol), acid chloride (2.2mg), (dicyclohexyl phosphino-) biphenyl (7mg) and Potassium monofluoride (344mg, the 5.8mmol) mixture in THF (4ml) heated 20 minutes in 120 ℃ in microwave.
The reaction mixture that merges batch A and B filters and filtrate concentrates in a vacuum.Resistates is by flash chromatography (SiO 2, the hexane solution of 0-5%EtOAc) and purifying, obtain the mixture of initial substance and coupled product.The NaOH (aqueous solution) that this mixture is dissolved in ethanol (10ml) and 2M (5ml) in, then reflux is 3 hours.Remove in a vacuum solvent, and resistates is allocated in the HCl aqueous solution of DCM and 2M.Water layer extracts with other DCM.Merge organic phase and vacuum concentration.Thick material is by the reverse-phase chromatography purifying on Horizon, and the aqueous solution wash-out with 5-100%MeCN obtains title compound, is white solid (367mg) (DN108121-170A2, GSK1869780A) .MS (ES): C 14H 9F 3O 2Calculated value 266; Measured value 265 (M-H +).
The description of D24
3-cyano group-4-(2-methyl-propyl) methyl benzoate (D24)
Figure G200880012733XD00441
Under argon gas; to 3-cyano group-4-{[(trifluoromethyl) alkylsulfonyl] the oxygen base } methyl benzoate (D19) (1.5g; 4.85mmol) in be added in bromination (2-methyl-propyl) zinc (48.5ml, 24.25mmol) in tetrahydrofuran (THF) (50ml).Then add in this solution dichloride 1,1 '-two (diphenylphosphino) ferrocene-palladium (II) methylene dichloride mixtures (0.355g, 0.485mmol), and will react reflux 6 hours.Then mixture water (2mL) cancellation by the Sai Lite diatomite filtration, washs with ethyl acetate.Remove in a vacuum solvent.Resistates is allocated in ethyl acetate (50mL) and water (50mL), and with organic phase dry (phase splitter), and remove in a vacuum solvent.Resistates is by the silica gel chromatography purifying, through 40 minutes isohexane eluant solutions with 0-15%EtOAc.Collect 2 batches of products, wherein a collection of is title compound, is colorless oil (233mg, 1.072mmol).δH(CDCl 3，400MHz)：8.28(1H，d)，8.15(1H，dd)，7.38(1H，d)，3.94，3H，s)，2.78(2H，d)，2.02(1H，m)，0.96(6H，d)。
The description of D25
3-cyano group-4-(2-methyl-propyl) phenylformic acid (D25)
Figure G200880012733XD00442
3-cyano group-4-(2-methyl-propyl) methyl benzoate (D24) (233mg, 1.072mmol) is dissolved in ethanol (4ml), and adds 2M aqueous sodium hydroxide solution (1ml, 2mmol).Stirring reaction 1 hour.The HCl aqueous solution (10mL) that adds 2M, and mixture extracts with methylene dichloride (20mL+10mL).Separate organic phase, and dry by phase splitter, merge, then remove in a vacuum solvent, obtain title compound, be white solid (203mg, 0.999mmol).MS (ES): C 12H 13NO 2Calculated value 203; Measured value 202 (M-H +).
The description of D26
5-formyl radical-2-{[(1S)-1-methyl-propyl] the oxygen base } benzonitrile (D26)
Figure G200880012733XD00451
(2S)-2-butanols (0.99g, 0.013mol) is dissolved in DMF (50ml), and solution is cooled to 0 ℃.Add wherein sodium hydride (60% dispersion in mineral oil, 1.54g, 0.036mol) in batches, add complete after mixture stirred 10 minutes in 0 ℃.Then add 2-fluoro-5-formyl radical benzonitrile (2.0g, 0.013mol), and reaction mixture is warmed to room temperature (slowly carrying out) in ice bath, then reaction mixture is in stirred overnight at room temperature.Then reaction mixture is cooled to 0 ℃, uses the salt solution cancellation, and with EtOAc (～25ml) dilution.Distribute mixture, and the organic moiety water (～30ml) extracting, then the organic layer of merging is evaporated to the dried crude product that obtains under the decompression by the post drying that is separated.Thick resistates purifying on the 40+MBiotage post, the hexanes mixtures wash-out of the EtOAc of use 20-50% obtains title compound (220mg), is white solid.MS (ES): C 12H 13NO 2Calculated value 203; Measured value 204 (MH +).
The description of D27
3-cyano group-4-{[(1S)-1-methyl-propyl] the oxygen base } phenylformic acid (D27)
To 5-formyl radical-2-{[(1S)-1-methyl-propyl] the oxygen base } benzonitrile (D26) (220mg; 1.08mmol) add sodium perborate tetrahydrate (334mg in solution in acetic acid (20ml); 2.17mmol), reaction mixture heated weekend in 50 ℃.Reaction mixture is concentrated in a vacuum.Add entry (～50ml), add EtOAc (～30ml) and separate each layer, water layer with EtOAc (～30ml) extraction 2 times, and with the organic phase vapourisation under reduced pressure that merges to doing, obtain title compound (245mg), be pale solid.MS (ES): C 12H 13NO 2Calculated value 219; Measured value 220 (MH +).
The description of D28
5-formyl radical-2-{[(1R)-1-methyl-propyl] the oxygen base } benzonitrile (D28).
Figure G200880012733XD00461
(2R)-2-butanols (0.99g, 0.013mol) is dissolved in DMF (50ml), and solution is cooled to 0 ℃.Add wherein sodium hydride in batches, 60% dispersion (1.54g, 0.036mol) in mineral oil, add complete after mixture stirred 10 minutes in 0 ℃.Then add 2-fluoro-5-formyl radical benzonitrile (2.0g, 0.013mol), and reaction mixture is warmed to room temperature (slowly carrying out) in ice bath, and reaction mixture is in stirred overnight at room temperature.Then reaction mixture is cooled to 0 ℃, uses the salt solution cancellation, and with EtOAc (～25ml) dilution.Separating mixture, and the organic moiety water (～30ml) extracting, the organic phase of merging is by the post drying that is separated, and vapourisation under reduced pressure is to the dried crude product that obtains.Thick resistates purifying on the 40+MBiotage post, the hexanes mixtures wash-out of the EtOAc of use 20-50%.Obtain title compound (310mg), be yellow oil.δH(d 6-DMSO，400MHz)：9.88(1H，s)，8.30(1H，s)，8.15(1H，d)，7.49(1H，d)，4.73-4.81(1H，m)，1.63-1.79(2H，m)，1.33(3H，d)，0.95(3H，t)。
The description of D29
3-cyano group-4-{[(1R)-1-methyl-propyl] the oxygen base } phenylformic acid (D29)
Figure G200880012733XD00462
To 5-formyl radical-2-{[(1R)-1-methyl-propyl] the oxygen base } benzonitrile (D37) (310mg; 1.53mmol) add sodium perborate tetrahydrate (471mg in solution in acetic acid (30ml); 3.05mmol), reaction mixture heated weekend in 50 ℃.Reaction mixture is concentrated in a vacuum, and add entry (～50ml), add EtOAc (～30ml), and separate each layer, water layer use again EtOAc (～30ml) extraction 2 times, and the organic phase vapourisation under reduced pressure that merges is pale solid to the dried title compound (315mg) that obtains.MS (ES): C 12H 13NO 2Calculated value 219; Measured value 220 (MH +).
The description of D30
5-chloro-6-[(1-methylethyl) oxygen base]-acidum nicotinicum 1-methyl ethyl ester (D30)
With 5-chloro-6-hydroxyl-acidum nicotinicum (3.00g, 17.28mmol) be suspended in toluene (75ml), and with silver carbonate (12.4g, 24.8mmol) and 2-iodopropane (6.9ml, 69.12mmol) process, and stirred in the dark 18 hours in room temperature.Filtering mixt, and evaporation filtrate obtains impure title compound (1.32g).δ H (d 6-DMSO) 1.31 (6H, d), 1.35 (6H, d), 5.13 (1H, septets), 5.40 (1H, septets), 8.22 (1H, d), 8.65 (1H, d).
The description of D30 (another kind of method)
5-chloro-6-[(1-methylethyl) oxygen base]-acidum nicotinicum 1-methyl ethyl ester (D30)
Figure G200880012733XD00472
With 5-chloro-6-hydroxyl-acidum nicotinicum (1g, 5.76mmol) be suspended in toluene (200ml), and with silver carbonate (3.97g, 14.40mmol) and 2-iodopropane (3.46ml, 34.6mmol) process, and stirred in the dark 3 days in room temperature.Add another part 2-iodopropane (3ml), and stirred 24 hours.Add EtOAc (200ml), and water (200ml) and saturated NaHCO 3(50ml) washing, then water (200ml) washing.
Through MgSO 4Drying, and evaporating solvent obtains the title compound of 1.0g, is clear, colorless oily matter, further just do not characterize and uses.
The description of D30 (another kind of method)
5-chloro-6-[(1-methylethyl) oxygen base]-acidum nicotinicum 1-methyl ethyl ester (D30)
Figure G200880012733XD00481
2-iodopropane (12.42ml, 124mmol) is joined 5-chloro-6-hydroxyl-acidum nicotinicum (being purchased from Fluka, 5.4g, 31.1mmol) and Ag 2CO 3(21.45g, 78mmol) is at CHCl 3In suspension (200ml), and reacted on stirring at room 3 days.Leach precipitation, and remove in a vacuum solvent and obtain title compound (4g, 47%), be yellow oil, it is not further purified and just is used for next step.MS (ES) C 12H 16ClNO 3Calculated value 257, measured value 258[M+H] +
The description of D31
5-chloro-6-[(1-methylethyl) oxygen base]-acidum nicotinicum (D31)
Figure G200880012733XD00482
5-chloro-6-[(1-methylethyl in Virahol (70ml) and water (35.0ml)) oxygen base]-acidum nicotinicum 1-methyl ethyl ester (D30) (1.6g, 6.21mmol) use 2N sodium hydroxide (6.21ml, 12.42mmol) process, and stir and obtained single product in 3 hours.Evaporation IPA also uses EtOAc (100ml) extraction product with the Glacial acetic acid acidifying.Through MgSO 4Dry and evaporating solvent obtains the title compound of 1.30g, is white solid.MS (ES) C 9H 10 35ClNO 3Calculated value 215; Measured value 214 (M-H +).
The description of D31 (another kind of method)
5-chloro-6-[(1-methylethyl) oxygen base]-acidum nicotinicum (D31)
Figure G200880012733XD00491
With sodium hydroxide (29.1ml, 58.2mmol) join 5-chloro-6-[(1-methylethyl) the oxygen base]-acidum nicotinicum 1-methyl ethyl ester (D30) (3g, 11.64mmol) in solution in methyl alcohol (25ml), and in stirred overnight at room temperature.Evaporating solvent, then the HCl with 2M neutralizes, and extracts with ether.Then through dried over mgso, and evaporation obtains impure white solid.Grind with DCM and obtain title compound, be the pure white solid.MS (ES) C 9H 10ClNO 3Calculated value 215, measured value 216[M+H] +
The description of D32
4-(1-tetrahydrobenzene-1-yl)-3-(trifluoromethyl) benzamide (D32)
Figure G200880012733XD00492
With 4-bromo-3-(trifluoromethyl) benzonitrile (being purchased) (1.2g, 4.80mmol), 1-tetrahydrobenzene-1-ylboronic acid (0.907g, 7.20mmol), sodium methylate (0.778g, 14.40mmol) and chlorination two (triphenylphosphines) close palladium (II) (0.337g, 0.480mmol) and join in anhydrous methanol (12mL), and mixture in microwave in 80 ℃ of heating 10 minutes.Reaction mixture is assigned in ethyl acetate (40mL) and water (40mL) then organic phase water (40mL) washing again.With the dry (MgSO of organic phase 4), filter and remove in a vacuum solvent.Crude product is by fast silica gel chromatogram method purifying, and the hexane solution wash-out with the 0-75% ethyl acetate obtains title compound, is white solid (1.02g).MS (ES): C 14H 14F 3NO calculated value 269; Measured value 270 (MH +).
The description of D33
4-cyclohexyl-3-(trifluoromethyl) benzamide (D33)
Figure G200880012733XD00501
4-(1-tetrahydrobenzene-1-yl)-3-(trifluoromethyl) benzamide (D32) (850mg, 3.16mmol) is dissolved in methyl alcohol (63ml), and uses H-Cube, use palladium/carbon in 40 ℃ of hydrogenations, flow velocity is 2mL/ minute.Remove in a vacuum solvent and obtain title compound, be white solid (822mg).MS (ES): C 14H 16F 3NO calculated value 271; Measured value 272 (MH +).
The description of D34
4-cyclohexyl-3-(trifluoromethyl) phenylformic acid (D34)
Figure G200880012733XD00502
To 4-cyclohexyl-3-(trifluoromethyl) benzamide (D33) (822mg, 3.03mmol) add potassium hydroxide (1.700g in solution in ethanol (40ml), 30.3mmol) and water (5ml), and reaction is heated to 90 ℃ of device temperatures, kept 3 hours, and in stirring at room 16 hours.Add another part potassium hydroxide (1.700g, 30.3mmol), and will react reflux 27 hours.Add again 5mL water, and reacting by heating 66 hours (spending weekend).Reaction mixture is concentrated in a vacuum, and resistates is allocated in ethyl acetate (25mL) and aqueous hydrochloric acid (2M, 25mL).Water layer is used ethyl acetate (25mL) extraction again, and with the dry (MgSO of the organic phase that merges 4), also removing in a vacuum solvent obtains title compound in filtration, is white solid (737mg).MS (ES): C 14H 15F 3O 2Calculated value 272; Measured value 271 (M-H +).
The description of D35
The 4-[(1-methylethyl) oxygen base]-3-(trifluoromethyl) phenylformic acid 1-methyl ethyl ester (D35)
Figure G200880012733XD00511
With 4-hydroxyl-3-(trifluoromethyl) phenylformic acid (being purchased) (450mg, 2.18mmol), 2-iodopropane (435 μ L, 4.36mmol) and salt of wormwood (603mg, 4.36mmol) at N, the mixture in N '-dimethyl formamide (40mL) and then adds 2-iodopropane (218 μ L in 70 ℃ of heating 4 hours, 2.18mmol), and continue heating 18 hours.The filtering inoganic solids, and wash with ethyl acetate.Filtrate is concentrated in a vacuum, and is allocated in ethyl acetate (150mL) and contains in the water (150mL) of some aqueous sodium hydroxide solutions.With organic layer dry (phase splitter), and vacuum concentration obtains thick title compound (704mg), is yellow oil.MS (ES): C 14H 17F 3O 3Calculated value 290; Measured value 291 (MH +).
The description of D36
The 4-[(1-methylethyl) oxygen base]-3-(trifluoromethyl) phenylformic acid (D36)
Figure G200880012733XD00512
To the 4-[(1-methylethyl) the oxygen base]-3-(trifluoromethyl) phenylformic acid 1-methyl ethyl ester (D35) (704mg, 2.43mmol) add aqueous sodium hydroxide solution (2M in mixture in ethanol (110mL), 12.2mL, 24.3mmol), and reacted reflux 1 hour.Mixture is concentrated in a vacuum, and resistates is allocated in ethyl acetate (100mL) and water (100mL), and with aqueous hydrochloric acid (2M, 13mL) acidifying.Water layer is used ethyl acetate (100mL) extraction again, and the dry also vacuum concentration of the organic layer that merges is obtained title compound, is yellow solid (563mg).MS (ES): C 11H 11F 3O 3Calculated value 248; Measured value 247 (M-H +).
The description of D37
3-chloro-4-(propoxy-) methyl benzoate (D37)
Figure G200880012733XD00521
4-hydroxyl-3-chloro benzoic ether (10g, 53.6mmol) is dissolved in DMF (110ml), then adds salt of wormwood (14.8g, 107.2mmol), just then adding-propyl iodide (10.4ml, 107.2mmol).Reaction is heated to 70 ℃ spends the night, filter, then with filtrate distribution in EtOAc and water.Separate organic layer, dry and evaporation obtains title compound, is yellow oil (12.37g).MS (ES) C 11H 13 35ClO 3Calculated value 228; Measured value 229 (MH +).
The description of D38
3-chloro-4-(propoxy-) phenylformic acid (D38)
Figure G200880012733XD00522
With 3-chloro-4-(propoxy-) methyl benzoate (D37) (RD108825-58-B1) (12.22g, 0.053mol) solution in ethanol (40ml) and the 2M NaOH aqueous solution (40ml) in 60 ℃ the heating 3 hours.Make reaction cooling, then spend weekend in room temperature.Reaction mixture is poured in the mixture of rare HCl aqueous solution and EtOAc.Separate organic layer, dry and evaporation obtains solid, it is ground with ether obtain title compound, is white solid (7.7g).MS (ES) C 10H 11 35ClO 3Calculated value 214; Measured value 213 (M-H +).
The description of D39
3-chloro-4-(1-pyrrolidyl) ethyl benzoate (D39)
Under argon gas with 4-bromo-3-chloro-benzoic acid ethyl ester (D16) (2.53g, 9.62mmol), tetramethyleneimine (1.03mL, 12.5mmol), sodium tert-butoxide (1.29g, 13.5mmol), BINAP (196mg, 0.29mmol) and three (dibenzalacetones) close the mixture heating up of two palladiums (0) (58mg, 0.10mmol) in toluene (120mL) and refluxed 3 hours.To react cooling and filter.Solid toluene wash, then concentrated filtrate.Resistates is allocated in ethyl acetate (150mL) and contains in the water (100mL) of sodium bicarbonate aqueous solution (50mL).With organic phase dry (phase splitter), and the concentrated orange that obtains.By the reverse-phase chromatography purifying, the aqueous solution wash-out with the 5-100% acetonitrile obtains title compound, is orange (600mg) with this oily matter.It contains some impurity.MS (ES): C 13H 16 35ClNO 2Calculated value 253; Measured value 254 (MH +).
Other title compounds (85mg) prepare by similarity method.
The description of D40
3-chloro-4-(1-pyrrolidyl) phenylformic acid (D40)
Figure G200880012733XD00541
Mixture in ethanol (15mL) was in 40 ℃ of heating 17 hours with 3-chloro-4-(1-pyrrolidyl) ethyl benzoate (D39) (685mg, 2.71mmol) and aqueous sodium hydroxide solution (2M, 1.36mL, 2.72mmol).Add again sodium hydroxide (1.36mL, 2.72mmol), and reheat reaction 8 hours.Enriched mixture, and resistates is dissolved in water (100mL).Then water is acidified to pH 6 with ethyl acetate (2x 100mL) washing.Form white solid and with its filtering, wash with water, and drying obtain title compound in vacuum drying oven, being pale solid (362mg).MS (ES): C 11H 12 35ClNO 2Calculated value 225; Measured value 226 (MH +).
The description of D41
2-(cyclopentyloxy)-5-formyl radical benzonitrile (D41)
Figure G200880012733XD00542
The solution of cyclopentanol (1.15g, 0.013mol) in DMF (50ml) is cooled to 0 ℃.Sodium hydride (1.54g, 0.016mol) is joined in this solution in batches, and add complete after mixture stirred 10 minutes in 0 ℃.Then add 3-cyano group-4-fluorobenzaldehyde (being purchased) (2.0g, 0.013mol), and mixture is warmed to room temperature and stirs and spend the night.Reaction mixture is cooled to 0 ℃, and adds salt solution, the mixture ethyl acetate (～25ml) dilute and distribute, the organic layer water (～30ml) washing, and by the post drying that is separated.Solution is concentrated in a vacuum, and resistates is by Biotage SP4, the 40M purifying, with the 20-60% ethyl acetate/hexane as elutriant.The fraction that contains product concentrates in a vacuum and obtains title compound (1.02g, 4.2mmol), is yellow liquid.MS (ES): calculated value C 13H 13NO 2215; Measured value 216 (MH +).
The description of D42
3-cyano group-4-(cyclopentyloxy) phenylformic acid (D42)
Figure G200880012733XD00551
In 50 ℃, add sodium perborate tetrahydrate (0.186g, 1.208mmol) in (D41) (0.13g, 0.604mmol) solution in acetic acid (20ml) in batches.Reaction mixture is spent the night 50 ℃ of stirrings.Continue to stir 48 hours in 50 ℃, sodium perborate tetrahydrate (0.093g, 0.604mmol) is joined in reaction mixture, and continue to stir 2 days, then with reaction mixture in room temperature standing 2 days.
Remove in a vacuum solvent, and with mixture be allocated in ethyl acetate (～20ml) and water (～20ml) in, (2x～15ml) the extraction of water layer ethyl acetate, and the organic layer that merges is the concentrated title compound (106mg that obtains in a vacuum, 0.435mmol), be yellow solid.MS (ES): calculated value C 13H 13NO 3231; Measured value 230 (M-H +).
The description of D43
5-[3-(1H-indazole-5-yl)-1,2,4-oxadiazole-5-yl]-the 2-[(1-methylethyl) the oxygen base] benzonitrile (D43)
With 3-cyano group-4-[(1-methylethyl) the oxygen base] (1.52g), EDAC (2.14g) and HOBt (1.51g) are dissolved in DMF (30ml) to phenylformic acid (can as preparation as described in WO2005/58848), then stir 5 minutes.Add N-hydroxyl-1H-indazole-5-carbonamidine (D2) (1.3g), and reaction is heated to 80 ℃, kept 6 hours, and cool overnight.Remove in a vacuum solvent, and crude product is allocated in EtOAc and sodium bicarbonate aqueous solution.Separate organic layer, drying also is evaporated to dried in a vacuum.The gained solid grinds with ether and ethyl acetate, with methyl alcohol and ether washing, obtains title compound (816mg), is pink solid.MS (ES): C 19H 15N 5O 2Calculated value 345; Measured value (MH +) 346
The description of D43 (another kind of method)
5-[3-(1H-indazole-5-yl)-1,2,4-oxadiazole-5-yl]-the 2-[(1-methylethyl) the oxygen base] benzonitrile (D43)
Figure G200880012733XD00561
With 3-cyano group-4-[(1-methylethyl) the oxygen base] Benzoyl chloride (D106) (545mg, 2.437mmol) and N-hydroxyl-1H-indazole-5-carbonamidine (D2) (685mg, 2.437mmol) be dissolved in acetonitrile (30ml), then add triethylamine (0.374ml, 2.68mmol).The gained mixture is in stirring at room 3 hours, then stirs 48 hours in 80 ℃, and is then cooling in room temperature.Leach the precipitation of formation and be dissolved in EtOAc, then washing with water, evaporate and use chromatogram purification [cyclohexane solution of 0-15%EtOAc] to obtain title compound (170mg, 17%), being pink solid.MS (ES): C 19H 15N 5O 2Calculated value 345; Measured value (MH +) 346
The description of D44 and D45
4-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-2H-indazole-2-yl] ethyl butyrate (D44) and 4-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-2H-indazole-2-yl] ethyl butyrate (D45)
Figure G200880012733XD00571
With 5-[3-(1H-indazole-5-yl)-1,2,4-oxadiazole-5-yl]-the 2-[(1-methylethyl) the oxygen base] benzonitrile (D43) (200mg, 0.58mmol) is dissolved in DMF (5ml), and adds Cs 2CO 3(567mg, 1.74mmol) and 4-bromo-butyric acid ethyl ester (0.166ml, 1.16mmol).Reaction is heated to 80 ℃, kept 12 hours, then cooling.Reaction mixture is allocated in water and EtOAc.Separate organic layer, dry also the evaporation in a vacuum obtains crude product, it is by flash chromatography (EtOAc/ sherwood oil 1: 6 to 1: 1) purifying, obtain 4-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] ethyl butyrate (D44) (151mg), be white solid.δ H (d 6-DMSO) 1.14 (3H, t), 1.39 (6H, d), 2.04-2.18 (2H, m), 2.31 (2H, t), 4.01 (2H, q), (4.51 2H, t), 4.99 (1H, septets), 7.57 (1H, d), 7.87 (1H, d), 8.08 (1H, dd), (8.29 1H, d), 8.43 (1H, dd), (8.52 1H, d), 8.57-8.58 (1H, m) .MS (ES): C 25H 25N 5O 4Calculated value 459; Measured value (MH +) 460.And RD108825-187A24-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl-1,2,4-oxadiazole-3-yl)-2H-indazole-2-yl] ethyl butyrate (D45) (66mg), be white solid.δ H (d 6-DMSO) 1.16 (3H, t), 1.37 (6H, d), 2.14-2.26 (2H, m), 2.32 (2H, t), 4.03 (2H, q), (4.51 2H, t), 4.99 (1H, septets), 7.57 (1H, d), 7.78-7.80 (1H, m), 7.89 (1H, dd), (8.42 1H, dd), 8.52 (1H, d), (8.57-8.58 1H, m), 8.62 (1H, s) .MS (ES) C 25H 25N 5O 4Calculated value 459; Measured value (MH +) 460.
The description of D46
[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] ethyl acetate (D46)
Figure G200880012733XD00581
With the method similar to (D44) from 5-[3-(1H-indazole-5-yl)-1,2,4-oxadiazole-5-yl]-the 2-[(1-methylethyl) the oxygen base] preparation of benzonitrile (D43) and ethyl bromoacetate.Thick material obtains title compound by grinding purifying with ether, is yellow solid.MS (ES) C 23H 21N 5O 4Calculated value 431; Measured value (MH +) 432.
The description of D47
3-(5-cyano group-1H-indazole-1-yl) ethyl propionate (D47)
Figure G200880012733XD00582
Add cesium carbonate (12.86g, 39.4mmol) in the 1H-indazole-solution of 5-formonitrile HCN (D1) (2.8g, 19.72mmol) in DMF (60ml), then add 3-ethyl bromide (3.8ml, 29.6mmol).Reaction mixture was in 80 ℃ of stirrings 3 hours, and is then cooling.Remove in a vacuum solvent, and with thick dispensed materials in EtOAc (x2) and water, merge organic phase, dry and evaporation.Crude product and methylbenzene azeotropic then by flash chromatography (EtOAc/ hexane 1: 3) purifying, obtain the pink solid of 3.75g, and it obtains title compound (3.2g) after grinding with hexane, is pink solid.MS (ES): C 13H 13N 3O 2Calculated value 243 measured value (MH +) 244.
The description of D47 (another kind of method)
3-(5-cyano group-1H-indazole-1-yl) ethyl propionate (D47)
Figure G200880012733XD00591
Add Cs in the 1H-indazole-solution of 5-formonitrile HCN (D1) (3.88g, 27.3mmol) in DMF (85ml) 2CO 3(17.8g, 54.6mmol) then adds 3-ethyl bromide (3.5ml, 27mmol) and 3-chloropropionate (3.3g, 24.2mmol).Reaction mixture was in 80 ℃ of stirrings 4 hours, and is then cooling.Remove in a vacuum solvent, and slightly dispensed materials in EtOAc and water, is separated organic layer, uses the salt water washing, dry and evaporation.Thick material and methylbenzene azeotropic then by flash chromatography (EtOAc/ hexane 1: 9 to 1: 1) purifying, obtain the yellow jelly of 3.84g.The MS data are as above-described embodiment.
The description of D47 (another kind of method)
3-(5-cyano group-1H-indazole-1-yl) ethyl propionate (D47)
Figure G200880012733XD00592
1H-indazole-5-formonitrile HCN (D1) (3.5g, 24.45mmol) is dissolved in DMF (60ml).Add cesium carbonate (15.93g, 48.9mmol) and 3-ethyl bromide (4.68ml, 36.7mmol).Mixture was in 80 ℃ of heating 4 hours.With the EtOAc dilution, and water (3x40ml) washing, through MgSO 4Drying, and evaporation.Mixture is by purified by flash chromatography, with 10-30% 10-20%EtOAc/ isohexane wash-out then, obtains (3.682g) the first isomer (yellow oil) and (1.509g) the second isomer (pink solid).Analyze the first isomer by 1H NMR and show that it is the 1-isomer that contains 2-isomer (approximately 20%).δ H (d 6-DMSO, 400MHz) 1.05 (3H, t), 2.95 (2H, t), 3.97 (2H, q), 4.69 (2H, t), 7.74 (1H, dd), 7.92 (1H, dt), 8.28 (1H, d), 8.38 (1H, dd) .MS (ES +): C 13H 13N 3O 2Calculated value 243; Measured value (MH +) 244.
The description of D48
The mixture (D48) of 4-(4-cyano group-2H-indazole-2-yl) ethyl butyrate and 4-(4-cyano group-1H-indazole-1-yl) ethyl butyrate
Figure G200880012733XD00601
With 4-bromo-butyric acid ethyl ester (0.390g, 1.998mmol), 1H-indazole-4-formonitrile HCN (0.143g, 0.999mmol) and cesium carbonate (0.976g, 3.00mmol) mixture in DMF (4ml) stirred and heat 90 minutes in 80 ℃.Cooling, dilute with ethyl acetate/water (every kind of 30ml), and organic phase 3x15ml water washing, dry (sal epsom), evaporate and pass through purified by flash chromatography, with 1: 2 ethyl acetate/hexane wash-out, obtaining 2 kinds of isomer that part is separated, merge after it is characterized, obtain the mixture (249mg) of title isomer.δ H (CDCl 3, 400MHz) for the 1-isomer: 1.24 (3H, t), 2.29 (4H, m), 4.12, (2H, q), 4.53 (2H, t), 7.45 (1H, dd), (7.54 1H, dd), 7.71 (1H, dd), 8.19 (1H, d). δ H (CDCl 3, 400MHz) for the 2-isomer: 1.25 (3H, t), 2.35 (4H, m), 4.14, (2H, q), 4.58 (2H, t), 7.33 (1H, dd), 7.54 (1H, dd), 7.96 (1H, dd), 8.15 (1H, d).
The description of D48 (another kind of method)
The mixture (D48) of 4-(4-cyano group-2H-indazole-2-yl) ethyl butyrate and 4-(4-cyano group-1H-indazole-1-yl) ethyl butyrate
Figure G200880012733XD00611
With 4-bromo-butyric acid ethyl ester (5.36ml, 37.4mmol), 1H-indazole-4-formonitrile HCN (2.68g, 18.72mmol) and cesium carbonate (18.30g, 56.2mmol) mixture in DMF (50ml) is in 80 ℃ of heating 90 minutes.Cooling, with the dilution of EtOAc/ water, and organic phase 3x30ml water washing, dry (sal epsom) and evaporation.By purified by flash chromatography, with 1: 3 wash-out of EtOAc/ hexane, obtain the mixture (4.7g) of title isomer, be yellow oil.MS (ES+): C 14H 15N 3O 2Calculated value 257; Measured value (MH +) 258.
The description of D49
The mixture (D49) of 3-(4-cyano group-2H-indazole-2-yl) ethyl propionate and 3-(4-cyano group-1H-indazole-1-yl) ethyl propionate
Figure G200880012733XD00612
With 3-ethyl bromide (1.808g, 9.99mmol), 1H-indazole-4-formonitrile HCN (0.715g, 4.99mmol) and cesium carbonate (4.88g, 14.98mmol) mixture at DMF (DMF) in (10ml) stirred and heat 2 hours in 80 ℃.Cooling, with ethyl acetate (40ml) and water (40ml) dilution, and organic phase 3x15ml water washing, dry (sal epsom), evaporate and pass through purified by flash chromatography, with 3: 7 ethyl acetate/hexane wash-outs, obtain the 1.19g colorless oil, its slow crystallization obtains white solid.NMR and LC/MS show that it is～3: 1 isomer mixture.MS (ES): C 13H 13N 3O 2Calculated value 243 measured value (MH +) 244.
The description of D50 and D51
4-(5-cyano group-1H-indazole-1-yl) ethyl butyrate (D50) and 4-(5-cyano group-2H-indazole-2-yl) ethyl butyrate (D51)
Figure G200880012733XD00621
Add cesium carbonate (9.1g, 27.9mmol) in the 1H-indazole-solution of 5-formonitrile HCN (D1) (1.93g, 13.59mmol) in DMF (45ml), then add 4-bromo-butyric acid ethyl ester (2.92ml, 20.4mmol).React on 80 ℃ of heating 6 hours.Most of DMF removes in a vacuum, and the gained material is allocated in ethyl acetate and water, organic phase salt water washing, and dry and evaporation obtains the 4g jelly.Thick material is by flash chromatography (using the hexane solution of 1: 4 EtOAc) purifying.Separate the top spot and obtain RD107973-39B1, it is accredited as 4-(5-cyano group-1H-indazole-1-yl) ethyl butyrate (D50).δ H (d 6-DMSO) 1.13 (3H, t), 2.04-2.13 (2H, m), 2.29 (2H, t), (3.99 2H, q), 4.51 (2H, t), 7.74 (1H, dd), (7.88 1H, d), 8.28 (1H, d), 8.41-8.44 (1H, m) .MS (ES): C 14H 15N 3O 2Calculated value 257 measured value (MH +) 258.
Separate the bottom spot and obtain title compound.δ H (d 6-DMSO) 1.15 (3H, t), 2.15-2.22 (2H, m), 2.31 (2H, t), (4.01 2H, q), 4.49-4.55 (2H, m), 7.48 (1H, dd), (7.78 1H, dd), 8.43-8.44 (1H, m), 8.67 (1H, d) .C 14H 15N 3O 2Calculated value 257 measured value (MH +) 258.
The description of D52
The 3-{5-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl } ethyl propionate (D52)
Figure G200880012733XD00631
3-(5-cyano group-1H-indazole-1-yl) ethyl propionate (D47) (1g, 4.11mmol) is dissolved in ethanol (30ml), and adds sodium bicarbonate (1.73g, 20.55mmol) and hydroxylamine hydrochloride (571mg, 8.22mmol).React on 50 ℃ of heated overnight, then cooling.Filter reaction mixture, and the resistates washing with alcohol.Filtrate is evaporated in a vacuum dried, and the thick title compound of gained (1.06g) is used for next step reaction.
The description of D52 (another kind of method)
The 3-{5-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl } ethyl propionate (D52)
3-(5-cyano group-1H-indazole-1-yl) ethyl propionate (D47) (1g, 4.11mmol) is dissolved in ethanol (30ml), and adds sodium bicarbonate (1.73g, 20.88mmol) and hydroxylamine hydrochloride (571mg, 8.22mmol).React on the room temperature placement and spend the night, then be heated to 50 ℃, kept 4 hours, then cool overnight.Filter reaction mixture, and the resistates washing with alcohol.The filtrate that contains ethanol is evaporated to dried in a vacuum, then grinds with ether and obtains title compound (764mg), is white solid, purity approximately 90%.MS (ES): C 13H 16N 4O 3Calculated value 276; Measured value (MH +) 277.
The description of D52 (another kind of method)
The 3-{5-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl } ethyl propionate (D52)
Figure G200880012733XD00641
With 3-(5-cyano group-1H-indazole-1-yl) ethyl propionate (D47) (3.84g, 15.8mmol) be dissolved in ethanol (100ml), and add sodium bicarbonate (6.6g, 79mmol), then add hydroxylamine hydrochloride (3.3g, 47.4mmol).Reaction is heated to 50 ℃, kept 11 hours altogether, then cool overnight.Filter reaction and resistates washing with alcohol.Filtrate is evaporated to dried in a vacuum, and the gained solid grinds with ether and obtain title compound (3.79g), is white solid.MS (ES): C 13H 16N 4O 3Calculated value 276; Measured value (MH +) 277.
The description of D52 (another kind of method)
The 3-{5-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl } ethyl propionate (D52)
3-(5-cyano group-1H-indazole-1-yl) ethyl propionate D47 (3.6g, 14.80mmol) is dissolved in ethanol (40ml), adds hydroxylamine hydrochloride (2.057g, 29.6mmol) and sodium bicarbonate (6.22g, 74.0mmol).Mixture is in 60 ℃ of heated overnight.Cooling, filter, and evaporation filtrate, being dissolved in EtOAc, water (3x20ml) washing is through MgSO 4Drying, and evaporation obtains (1.75g) product again.Reclaim again the white crystal of 0.290g from rotatory evaporator.MS (ES +): C 13H 16N 4O 3Calculated value 276; Measured value (MH +) 277.
The description of D53
The 4-{5-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl } ethyl butyrate (D53)
Figure G200880012733XD00651
Title compound is by preparing to the described similar method of D50 RD107973-039B1.MS (ES): C 14H 18N 4O 3Calculated value 290; Measured value (MH +) 291.
The description of D54
The 4-{5-[(hydroxyl amino) (imino-) methyl]-2H-indazole-2-yl } ethyl butyrate (D54)
Figure G200880012733XD00652
Title compound is by preparing to the described similar method of 4-(5-cyano group-2H-indazole-2-yl) ethyl butyrate (D51).MS (ES): C 14H 18N 4O 3Calculated value 290; Measured value (MH +) 291.
The description of D55
The 3-{4-[(hydroxyl amino) (imino-) methyl]-2H-indazole-2-yl } ethyl propionate and 3-{4-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl } mixture (D55) of ethyl propionate
Figure G200880012733XD00661
With 3-(4-cyano group-2H-indazole-2-yl) ethyl propionate and 3-(4-cyano group-1H-indazole-1-yl) ethyl propionate (D49) (1.2g, 4.938mmol) (sample is the mixture of～3: 11 and 2-isomer), hydroxylamine hydrochloride (0.686g, 9.87mmol) and the mixture of sodium bicarbonate (2.072g, 24.66mmol) in ethanol (15ml) in 50 ℃ the heating 3 hours.Cross filter solid, and filtrate is evaporated to dried obtaining (1.4g) colorless oil.LCMS and NMR conclusive evidence is the mixture of～3: 1 isomer.MS (ES): C 13H 16N 4O 3Calculated value 276; Measured value (MH +) 277.
The description of D56
The 4-{4-[(hydroxyl amino) (imino-) methyl]-2H-indazole-2-yl } ethyl butyrate and 4-{4-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl } mixture (D56) of ethyl butyrate
Figure G200880012733XD00662
with 4-(4-cyano group-2H-indazole-2-yl) ethyl butyrate and 4-(4-cyano group-1H-indazole-1-yl) ethyl butyrate (D48) (0.249g, 0.968mmol) (sample is the mixture of～3: 21 and 2-isomer), hydroxylamine hydrochloride (0.135g, 1.936mmol) and sodium bicarbonate (0.407g, 4.84mmol) mixture in ethanol (5ml) stirred and heat 90 minutes in 80 ℃, then use ethyl acetate (30ml)/water (50ml) dilution, organic phase dry (sal epsom) and evaporation obtain title mixture (270mg), be colourless jelly.LC/MS is indicated as the mixture of the isomer of 3: 2 with a small amount of impurity.MS (ES): C 14H 18N 4O 3Calculated value 290; Measured value (MH +) 291.
The description of D56 (another kind of method)
The 4-{4-[(hydroxyl amino) (imino-) methyl]-2H-indazole-2-yl } ethyl butyrate and 4-{4-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl } mixture (D56) of ethyl butyrate
Figure G200880012733XD00671
With 4-(4-cyano group-2H-indazole-2-yl) ethyl butyrate and 4-(4-cyano group-1H-indazole-1-yl) ethyl butyrate (D48) (4.7g, 9.13mmol) (sample be 1 and the 2-isomer～2: 1 mixtures), hydroxylamine hydrochloride (2.54g, 36.5mmol) and the suspension of sodium bicarbonate (7.67g 91mmol) in ethanol (50ml) in 50 ℃ the heating 3 hours, cooling and the evaporation, then be allocated in EtOAc/ water.With organic layer dry (sal epsom), evaporation also obtains title mixture (4g) with the ether grinding, is white solid.NMR and LCMS are indicated as the isomer mixture of 2: 1.MS (ES): C 14H 18N 4O 3Calculated value 290; Measured value (MH+) 291.
The description of D57
3-(5-{5-[3-chloro-4-(oxyethyl group) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) ethyl propionate (D57).
Figure G200880012733XD00672
With the 3-{5-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl } ethyl propionate (D52) (265 mg, 0.96mmol) and 3-chloro-4-(oxyethyl group) phenylformic acid (D7) (192mg, 0.96mmol) stir together in 80 ℃ in DMF (5ml), and add EDAC (203mg) and HOBt (142mg).When the judgement reaction is completed (passing through lcms analysis), it is cooling, and reaction mixture is allocated in EtOAc and sodium bicarbonate aqueous solution.Separate organic layer, and wash with water, then dry and evaporation is ground with ethanol and is obtained crude product.By flash chromatography (EtOAc/ hexane 1: 3 to 1: 1) purifying, obtain title compound (87mg), be white solid.MS (ES): C 22H 21 35ClN 4O 4Calculated value 440 measured value (MH +) 441.
The description of D58
3-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] ethyl propionate (D58)
Figure G200880012733XD00681
With the 3-{5-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl } ethyl propionate (D52) (265mg, 0.96mmol), 3-cyano group-4-[(1-methylethyl) oxygen base] phenylformic acid (can according to the described preparation of WO2005/58848) (197mg, 0.96mmol), EDAC (203mg) and HOBt (142mg) stirred 2 hours in 80 ℃ in DMF (5ml).Spend weekend in stirring at room, and then in 80 ℃ of heating 4 hours.Reaction mixture is cooling, then be allocated in EtOAc and sodium bicarbonate.Separate organic layer, use the salt water washing, drying also is evaporated to dried in a vacuum.Thick material obtains title compound (214mg) by purified by flash chromatography, is white solid.MS (ES): C 24H 23N 5O 4Calculated value 445; Measured value (MH +) 446.
The description of D58 (another kind of method)
3-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] ethyl propionate (D58)
Figure G200880012733XD00691
With 3-cyano group-4-[(1-methylethyl) the oxygen base] phenylformic acid (can prepare according to described in WO2005/58848) (371mg, 1.81mmol), EDAC (547mg, 2.85mmol) and the mixture of HOBt (437mg, 2.85mmol) in DMF (5ml) placed 10 minutes in room temperature.Add the 3-{5-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl } ethyl propionate (D52) (500mg, 1.81mmol), then place in room temperature and spend the night.Add EDAC (200mg, 1.04mmol), and placed 24 hours.Mixture is placed in room temperature every night in 80 ℃ of heating 2 days.Mixture is again in 80 ℃ of heating 4 hours.Add EtOAc (50ml), and water (60ml), saturated NaHCO 3(60ml) and water (60ml) washing.With the dry (MgSO of EtOAc layer 4), and evaporating solvent.Grind with ethanol and obtain title compound (523mg), be beige solid.MS (ES): C 24H 23N 5O 4Calculated value 445; Measured value (MH +) 446.
The description of D59
3-(5-{5-[4-[(1-methylethyl) oxygen base]-3-(trifluoromethyl) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) ethyl propionate (D59)
With the 4-[(1-methylethyl) the oxygen base]-3-(trifluoromethyl) phenylformic acid (D36) (100mg, 0.4mmol) be dissolved in DMF (5ml), then add EDAC (115mg, 0.6mmol) and HOBt (81mg, 0.6mmol).Stirred the mixture 30 minutes, and then added the 3-{5-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl } ethyl propionate (D52) (111mg, 0.40mmol), and react on 80 ℃ of heated overnight.Add again EDAC (50mg), and reaction reheats 4 hours.Add again EDAC (20mg), and will react heated overnight.Reaction mixture is joined in the mixture of ethyl acetate and sodium bicarbonate, separate organic layer, and water layer is used ethyl acetate extraction again.Then the organic layer water that merges uses the salt water washing, and dry and evaporation obtains crude product.This material is by flash chromatography (SiO 2) purifying, obtain title compound (82mg).MS (ES): C 24H 23F 3N 4O 4Calculated value 488; Measured value (MH +) 489.
The description of D60
4-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] ethyl butyrate (D60)
Figure G200880012733XD00702
with the 4-{4-[(hydroxyl amino) (imino-) methyl]-2H-indazole-2-yl } ethyl butyrate and 4-{4-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl } mixture (the D56) (0.27g of ethyl butyrate, 0.930mmol) (mixture of 1 and the 2-isomer of 3: 2), 3-chloro-4-[(1-methylethyl) oxygen base] phenylformic acid (D3) (0.200g, 0.930mmol), EDC (0.196g, 1.023mmol) and HOBT (0.157g, 1.023mmol) at N, stirred 16 hours in 80 ℃ in dinethylformamide (5ml), then stirred 2 hours in 120 ℃.To react cooling, with ethyl acetate (40ml) and water (40ml) dilution, and organic phase 3x15ml water washing, dry (sal epsom) and evaporation.Thick material with 3: 7 ethyl acetate/hexane wash-outs, obtains two kinds of products by purified by flash chromatography, and obtains the top spot of 66mg and the bottom spot of 36mgs with ether/hexane grinding in 1: 1.The top spot is the indazole that required 1-replaces.δH(CDCl 3，400MHz)：1.24(3H，t)，1.46(6H，d)，2.31(4H，m)，4.12(2H，q)，4.55(2H，t)，4.73(1H，m)，7.08(1H，d)，7.52(1H，dd)，7.62(1H，d)，8.07(1H，dd)，8.11(1H，dd)，8.29(1H，d)，8.70(1H，d)。
The description of D60 (another kind of method)
4-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] ethyl butyrate (D60)
N2665-84-A1
Figure G200880012733XD00711
with the 4-{4-[(hydroxyl amino) (imino-) methyl]-2H-indazole-2-yl } ethyl butyrate and 4-{4-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl } mixture (the D56) (4g of ethyl butyrate, 13.78mmol), 3-chloro-4-[(1-methylethyl) oxygen base] phenylformic acid (D3) (2.96g, 13.78mmol), EDC (2.91g, 15.16mmol) and HOBT (2.321g, 15.16mmol) solution in DMF (40ml) formed intermediate in 3 hours in stirring at room, then heat 2 hours with Cheng Huan in 120 ℃.Solution is cooling, with EtOAc dilution, water 3x washing, dry (sal epsom) and evaporation.By purified by flash chromatography, with the EtOAc/ isohexane wash-out of 1: 3.The fraction evaporation that merges is obtained title compound (1.5g), be white solid.MS (ES): C 24H 25 35ClN 4O 4Calculated value 468; Measured value (MH +) 469. δ H (CDCl 3, 400MHz): 1.24 (3H, t), 1.46 (6H, d), (2.31 4H, m), 4.12 (2H, q), 4.55 (2H, t), 4.73 (1H, m), 7.08 (1H, d), (7.52 1H, dd), 7.62 (1H, d), (8.06 1H, dd), 8.10 (1H, dd), (8.29 1H, d), 8.70 (1H, d).
The description of D61
3-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] ethyl propionate (D61)
Figure G200880012733XD00721
with the 3-{4-[(hydroxyl amino) (imino-) methyl]-2H-indazole-2-yl } ethyl propionate and 3-{4-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl } mixture (the D55) (700mg of ethyl propionate, 1.27mmol) (mixture of 1 and the 2-isomer of 3: 1), 3-chloro-4-[(1-methylethyl) oxygen base] phenylformic acid (D3) (0.544g, 2.53mmol), EDC (0.534g, 2.79mmol) and HOBT (0.427g, 2.79mmol) at N, heated 2 hours in 120 ℃ in dinethylformamide (10ml), cooling, dilute with ethyl acetate (60ml), and water (3x20ml) washing.With the dry (MgSO of organic layer 4), evaporate and pass through purified by flash chromatography, the EtOAc/ hexane wash-out with 1: 4 obtains title compound (380mg), is white solid.δ H (CDCl 3, 400MHz): 1.17 (3H, t), 1.46 (6H, d), 3.02 (2H, t), 4.10 (2H, q), 4.70-4.77 (3H, m), 7.08 (1H, d), 7.53 (1H, dd), 7.69 (1H, d), 8.05-8.12 (2H, m), 8.29 (1H, d), 8.71 (1H, d) .MS (ES): C 23H 23 35ClN 4O 4Calculated value 454; Measured value (MH +) 455.
Following compound is by chemical process preparation similar to the aforementioned embodiment.In some cases, carboxylic acid, EDAC and HOBt stir together, then add amidoxim.In some cases, then reaction mixture heats in stirring at room.Except as otherwise noted, reaction is carried out in DMF at the temperature of 80-120 ℃.In some cases, then add EDAC and/or HOBt.In the situation of D91, then add carboxylic acid.Product is by following method purifying: flash chromatography, grinding, MDAP, reverse-phase chromatography, use ethanol precipitate product from reaction mixture, or the combination of these methods.
Numbering Structure Title Precursor MS
[0731]
Figure G200880012733XD00731
Figure G200880012733XD00751
Figure G200880012733XD00761
Figure G200880012733XD00771
Figure G200880012733XD00791
Figure G200880012733XD00801
The description of D96
3-(4-cyano group-1H-indazole-1-yl) ethyl propionate (D96)
Figure G200880012733XD00811
With 1H-indazole-4-formonitrile HCN (0.716g, 5mmol), 3-ethyl bromide (1.276ml, 10.00mmol) and cesium carbonate (4.90g, 15.00mmol) join N, dinethylformamide (DMF) (20ml) in, and with mixture heating up to 90 ℃, kept 80 minutes.Mixture is cooling, then use the mixture extraction of ethyl acetate and water.The organic moiety water washing of 3x30ml, dry (MgSO 4), filter and evaporate.Crude mixture is joined on the Biotage post, and with the ethyl acetate/petroleum ether wash-out of 1: 3, obtain the title compound of 0.95g.δH(CDCl 3，400MHz)1.17(3H，t)，3.01(2H，t)，4.08(2H，q)，4.71(2H，t)，7.46(1H，dd)，7.54(1H，dd)，7.81(1H，d)，8.21(1H，d)。
The description of D97
The 3-{4-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl } ethyl propionate (D97)
With 3-(4-cyano group-1H-indazole-1-yl) ethyl propionate (D96) (0.95g, 3.91mmol), hydroxylamine hydrochloride (1.086g, 15.62mmol), and sodium bicarbonate (3.28g, 39.1mmol) heated 3 hours in 50 ℃ in ethanol (20ml).Filtering mixt and evaporation obtain title compound (760mg), are white powder.MS (ES): C 13H 16N 4O 3Calculated value 276; Measured value (MH +) 277.
The description of D98
3-bromo-PA ethyl ester (D98)
Figure G200880012733XD00821
3-bromo-PA (being purchased from Fluka) (1g, 5.52mmol) is dissolved in thionyl chloride (5ml, 68.5mmol), and the gained mixture refluxed 2 hours, then be cooled to room temperature and vacuum concentration.Resistates is dissolved in ethanol (10ml), and in stirring at room 30 minutes.Remove in a vacuum most of solvent, resistates is dissolved in EtOAc, and with the NaOH aqueous solution of 1M, salt water washing 2 times, the dry and concentrated title compound (0.939g, 81%) that obtains, be light yellow oil, it is not further purified and just is used for next step.
The description of D99
4-bromo-2,2-dimethyl butyrate acetoacetic ester (D99)
Figure G200880012733XD00822
In room temperature, hydrogen bromide is blown in 3,3-dimethyl dihydro-2 (3H)-furanone (1g, 8.76mmol) approximately 5 hours, and the gained mixture is placed in room temperature and is spent the night.Then nitrogen is blown in mixture.Thick acid is dissolved in DCM (10ml), and processes with oxalyl chloride (2.3ml, 26.3mmol) and DMF (1), and the gained mixture spends weekend in stirring at room, then concentrated in a vacuum.Resistates is processed with ethanol (10ml), and then the gained mixture is poured into water in stirring at room 1 hour.Water extracted with diethyl ether 2 times, and ethereal solution salt water washing, dry and vacuum concentration obtains title compound (949mg, 49%), is very shallow yellow oil, and it is not further purified and just is used for next step.
The description of D100 and D101
3-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-2,2-ethyl dimethyl (D100), N6128-6-A4 and 3-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-2H-indazole-2-yl]-PA ethyl ester (D101)
Figure G200880012733XD00831
5-in DMF (2ml) (5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole (D4) (122mg, 0.344mmol) use 3-bromo-2,2-ethyl dimethyl (D98) (79mg, 0.378mmol) and Cs 2CO 3(134mg, 0.413mmol) processes, and the gained mixture spends the night in 80 ℃ of stirrings.Add again 3-bromo-PA ethyl ester (D98) (29mg, 0.4mmol) and Cs 2CO 3(56mg, 0.5mmol), and then the gained mixture is cooled to room temperature and vacuum concentration in 80 ℃ of stirrings 6 hours.Resistates is dissolved in EtOAc, and then the organic phase water uses the salt water washing, dry and vacuum concentration.Resistates is by fast silica gel chromatogram method [gradient: ethyl acetate/hexanaphthene] purifying, obtain 3-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-2,2-ethyl dimethyl (D100) (92mg, 55%) and 3-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl-1,2,4-oxadiazole-3-yl)-2H-indazole-2-yl]-2,2-ethyl dimethyl (D101) (49mg, 30%).MS (ES) C 25H 27ClN 4O 4Calculated value 482, measured value 483[M+H] +
The description of D102
3-(4-cyano group-1H-indazole-1-yl)-PA ethyl ester (D102)
Figure G200880012733XD00841
With the 1H-indazole in dry DMF (3ml)-4-formonitrile HCN (being purchased the Ltd. from Insight ChemicalSolutions) (500mg, 3.49mmol), 3-bromo-PA ethyl ester (D98) (803mg, 3.84mmol) and Cs 2CO 3(1.36g, 4.19mmol) is heated to 80 ℃ and spends the night, and then heats 7 hours.Then reaction mixture is allocated in EtOAc and H 2In O.Separates two, and organic phase is used H again 2O, salt water washing, the dry and concentrated yellow oil that obtains.Resistates obtains title compound (356mg, 37%) by fast silica gel chromatogram method [gradient: ethyl acetate/hexanaphthene] purifying, is crystalline material.MS (ES) C 15H 17N 3O 2Calculated value 271, measured value 272[M+H] +
The description of D102 (another kind of method)
3-(4-cyano group-1H-indazole-1-yl)-2,2--ethyl dimethyl (D102)
Figure G200880012733XD00842
Under nitrogen, with 1H-indazole-4-formonitrile HCN (being purchased the Ltd. from Insight Chemical Solutions, 298mg, 1.957mmol) at anhydrous N, dinethylformamide (DMF) is (1ml) middle to be processed with NaH (94mg, 2.348mmol).Then the solution of 3-bromo-PA ethyl ester (D98) (450mg, 2.153mmol) in DMF (2ml) is joined in this auburn mixture.Reaction mixture is heated to 80 ℃ spends the night, then concentrated in a vacuum.Resistates is dissolved in EtOAc, and then the organic phase water uses the salt water washing, the dry and concentrated brown jelly that obtains.Resistates obtains title compound (217mg, 86%) by fast silica gel chromatogram method [gradient: ethyl acetate/hexanaphthene] purifying, is yellow oil.MS (ES) C 15H 15N 3O 2Calculated value 271, measured value 272[M+H] +
The description of D103
The 3-{4-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl }-PA ethyl ester (D103)
Figure G200880012733XD00851
With 3-(4-cyano group-1H-indazole-1-yl)-PA ethyl ester (D102) (348mg, 1.28mmol), hydroxylamine hydrochloride (1.087g, 10.26mmol), NaHCO 3(1.078g, 12.83mmol) in 65 ℃ of heating 24 hours, then spends weekend in room temperature in MeOH (20ml).Mixture filters, and is concentrated, with water treatment and vigorous stirring 10 minutes, filters and drying obtains colorless solid, obtains title compound (373mg, 90%) in 45 ℃ of dried overnight in a vacuum.MS (ES) C 15H 20N 4O 3Calculated value 304, measured value 305[M+H] +
The description of D104
3-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-PA ethyl ester (D104)
With 3-chloro-4-[(1-methylethyl) the oxygen base] phenylformic acid (D3) (245mg, 1.143mmol), the 3-{4-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl }-2,2-ethyl dimethyl (D103) (348mg, 1.143mmol) mixture in DMF (5ml) in room temperature with N-ethyl-N-(1-methylethyl)-2-propylamine (0.240ml, 1.372mmol) process, then use HATU (522mg, 1.372mmol) to process.Then the gained mixture spends the night in 80 ℃ of stirrings, and then stirred 4 hours in stirring at room 1 hour, then is cooled to room temperature and vacuum concentration.Resistates is dissolved in EtOAc, and organic phase H 2Then O uses the salt water washing 2 times, dry and vacuum concentration.Resistates obtains title compound (233mg, 39%) by fast silica gel chromatogram method [gradient: ethyl acetate/hexanaphthene] purifying, is the paste solid.MS (ES) C 25H 27ClN 4O 4Calculated value 482, measured value 483[M+H] +
The description of D105
3-[4-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-PA ethyl ester (D105)
Figure G200880012733XD00861
With 5-chloro-6-[(1-methylethyl) the oxygen base]-acidum nicotinicum (D31) (70.9mg, 0.329mmol), triethylamine (0.092ml, 0.657mmol), HOBT (60.4mg, 0.394mmol) and EDC (76mg, 0.394mmol) 3-{4-[(hydroxyl amino then) (imino-) methyl]-1H-indazole-1-yl }-2,2-ethyl dimethyl (D103) (100mg, 0.329mmol) be dissolved in DMF (DMF) (5ml) in.The gained mixture spends the night in 50 ℃ of stirrings, then is cooled to room temperature, and dilutes with EtOAc.Organic phase H 2The O washing, dry and vacuum concentration.Resistates obtains title compound (30mg, 17%) by fast silica gel chromatogram method [15-40% EtOAc/ hexanaphthene] purifying, is light yellow solid.MS (ES) C 24H 26ClN 5O 4Calculated value 483, measured value 484[M+H] +
The description of D106
3-cyano group-4-[(1-methylethyl) oxygen base] Benzoyl chloride (D106)
Figure G200880012733XD00871
With oxalyl chloride (0.224ml, 2.56mmol) join the 3-cyano group of stirring-4-[(1-methylethyl) the oxygen base] (it can prepare described in WO2005/58848 phenylformic acid, 500mg, 2.437mmol) in solution in DCM (10ml), then add DMF (20 μ l), and the gained mixture was in stirring at room 2 hours.The solvent evaporation is obtained title compound (545mg, 100%), be the olive-green solid, it is not further purified and just is used for next step.
The description of D107
3-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl]-PA ethyl ester (D107)
Figure G200880012733XD00872
To 5-[3-(1H-indazole-5-yl)-1,2,4-oxadiazole-5-yl]-the 2-[(1-methylethyl) the oxygen base] benzonitrile (D43) (85mg, 0.246mmol) at N, add 3-bromo-2 in the solution of dinethylformamide (DMF) in (5ml), 2-ethyl dimethyl (D98) (103mg, 0.492mmol) and Cs 2CO 3(120mg, 0.369mmol).Then the gained mixture is cooled to room temperature, and dilutes with EtOAc in 80 ℃ of stirrings 48 hours.Organic phase washes with water, through MgSO 4Dry also vacuum concentration.Resistates obtains title compound (20mg, 16%) by fast silica gel chromatogram method [30-50% EtOAc/ tetrahydrobenzene] purifying, is yellow solid.MS (ES) C 26H 27N 5O 4Calculated value 473, measured value 474[M+H] +
The description of D108
4-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-2,2-dimethyl butyrate acetoacetic ester (D108)
Figure G200880012733XD00881
title compound uses and synthesizes 3-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-2, the similar method of 2-ethyl dimethyl (D107), from 5-[3-(1H-indazole-5-yl)-1,2,4-oxadiazole-5-yl]-the 2-[(1-methylethyl) the oxygen base] benzonitrile (D43), use 4-bromo-2,2-dimethyl butyrate acetoacetic ester (D99) replaces 3-bromo-2 as alkylating agent, 2-ethyl dimethyl (D98) and obtaining, productive rate is 31% (50mg).MS (ES) C 25H 25N 5O 4Calculated value 459, measured value 460[M+H] +
The description of D109
3-[4-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-PA ethyl ester (D109)
Figure G200880012733XD00882
With 3-cyano group-4-[(1-methylethyl) the oxygen base] Benzoyl chloride (D106) (0.398mmol) in acetonitrile (3.00ml) in room temperature 3-{4-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl-2,2-ethyl dimethyl (D103) (101mg, 0.332mmol) process, then use triethylamine (0.069ml, 0.498mmol) to process.The gained yellow solution was in stirring at room 1 hour, and backflow is spent the night, then 24 hours and again spend the night and then be cooled to room temperature and vacuum concentration.Resistates obtains title compound (107mg, 68%) by fast silica gel chromatogram method [gradient: ethyl acetate/hexanaphthene] purifying, is colorless solid.MS (ES) C 26H 27N 5O 4Calculated value 473, measured value 474[M+H] +
The description of D110
5-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-oxadiazole-3-yl)-1H-indazole (D110), N6043-42
With 5-chloro-6-[(1-methylethyl) the oxygen base]-acidum nicotinicum (D31) (350mg, 1.623mmol), triethylamine (328mg, 452 μ l, 3.25mmol), hydroxy benzotriazole hydrate (298mg, 1.95mmol), EDC (373mg, 1.95mmol) and the N-hydroxyl-1H-indazole-mixture of 5-carbonamidine (D2) (430mg, 2.43mmol) in DMF (5ml) stirred 4 days in 50 ℃.Reaction mixture is cooled to room temperature, and dilutes with EtOAc (25ml).The saturated NaHCO of organic layer 3The aqueous solution, water, then salt water washing.The dry also vacuum concentration of organic phase is obtained title compound (334mg, 58%), be the light red solid, it is not further purified and just is used for next step.MS (ES) C 17H 14ClN 5O 2Calculated value 355, measured value 356[M+H] +
The description of D111
3-[5-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-PA ethyl ester (D111)
With 5-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-oxadiazole-3-yl)-1H-indazole (D110) (335mg, 0.94mmol), 3-bromo-PA ethyl ester (D98) (197mg, 0.94mmol) and K 2CO 3(130mg, 0.94mmol) mixture in DMF (5ml) stirred 48 hours in 80 ℃.Then reaction mixture is cooled to room temperature, and is allocated in EtOAc and water (30ml 1: 1).Separate organic phase, through MgSO 4Dry also vacuum concentration.Resistates obtains title compound (100mg, 22%) by fast silica gel chromatogram method [20% ethyl acetate/hexanaphthene] purifying, is colorless solid.MS (ES) C 24H 26ClN 5O 4Calculated value 483, measured value 484[M+H] +
The description of D112
4-(4-bromo-1H-indazole-1-yl)-2,2-dimethyl butyrate acetoacetic ester (D112)
Figure G200880012733XD00902
In room temperature, add Cs in the solution of 4-bromo-1H-indazole (being purchased the Ltd. from Insight Chemical Solutions, 826mg, 4.19mmol) in DMF (10ml) 2CO 3(1.37g, 4.19mmol) and 4-bromo-2,2-dimethyl butyrate acetoacetic ester (D99) (1.40g, 6.29mmol), and then the gained mixture is cooled to room temperature, and dilutes with AcOEt in 80 ℃ of stirrings 24 hours.Organic phase washes with water, through MgSO 4Drying, and vacuum concentration.Resistates obtains title compound (600mg, 40%) by fast silica gel chromatogram method [15-25% EtOAc/ hexanaphthene] purifying, is orange.MS (ES) C 15H 19 81BrN 2O 2Calculated value 340, measured value 341[M+H] +
The description of D113
4-(4-cyano group-1H-indazole-1-yl)-2,2-dimethyl butyrate acetoacetic ester (D113), N6375-40
In room temperature, to 4-(4-bromo-1H-indazole-1-yl)-2,2-dimethyl butyrate acetoacetic ester (D112) (600mg, 1.77mmol) and Zn (CN) 2Add Pd (PPh in (207mg, 1.77mmol) solution in DMF (5ml) 3) 4(204mg, 0.18mmol), and the gained mixture stirred weekend in 100 ℃, reheated 5 hours, then was cooled to room temperature, and diluted with AcOEt.Organic phase is by the Sai Lite diatomite filtration, and then water uses the salt water washing, dry and vacuum concentration.Resistates obtains title compound (200mg, 36%) by fast silica gel chromatogram method [10-20%EtOAc/ hexanaphthene] purifying, is colorless oil.MS (ES) C 16H 19N 3O 2Calculated value 285, measured value 286[M+H] +
The description of D114
The 4-{4-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl }-2,2-dimethyl butyrate acetoacetic ester (D114)
Figure G200880012733XD00912
The 4-{4-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl }-2,2-dimethyl butyrate acetoacetic ester (D114) with synthetic 3-{4-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl-2, the similar method of 2-ethyl dimethyl (D103) is from 4-(4-cyano group-1H-indazole-1-yl)-2,2-dimethyl butyrate acetoacetic ester (D113) obtains, obtain 200mg, productive rate 85%.MS (ES) C 16H 22N 4O 3Calculated value 318, measured value 319[M+H] +
The description of D115
4-[4-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-2,2-dimethyl butyrate acetoacetic ester (D115)
Figure G200880012733XD00921
4-[4-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-2,2-dimethyl butyrate acetoacetic ester (D115) with synthetic 5-[3-(1H-indazole-5-yl)-1,2,4-oxadiazole-5-yl]-the 2-[(1-methylethyl) the oxygen base] the similar method of the another kind of method of benzonitrile (D43) is from 3-cyano group-4-[(1-methylethyl) the oxygen base] Benzoyl chloride (D106) and 4-{4-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl }-2,2-dimethyl butyrate acetoacetic ester (D114) obtains.120mg, productive rate 74%.MS (ES) C 27H 29N 5O 4Calculated value 487, measured value 488[M+H] +
The description of D116
4-[4-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-2,2-dimethyl butyrate acetoacetic ester (D116)
In room temperature, with 5-chloro-6-[(1-methylethyl) the oxygen base]-acidum nicotinicum (D31) (67.7mg, 0.314mmol), then triethylamine (88 μ l, 0.628mmol), then HOBT (57.7mg, 0.377mmol), EDC (72.3mg, 0.377) and the 4-{4-[(hydroxyl amino) (imino-) methyl]-1H-indazole-1-yl }-2,2-dimethyl butyrate acetoacetic ester (D114) (100mg, 0.314mmol) is dissolved in DMF (4ml), then stirs 5 days in 50 ℃.Mixture is cooled to room temperature, and dilutes with EtOAc.Organic phase washes with water, dry and vacuum concentration.Resistates obtains title compound (110mg, 67%) by fast silica gel chromatogram method [15-40%EtOAc/ hexanaphthene] purifying, is light yellow solid.MS (ES) C 25H 28ClN 5O 4Calculated value 497, measured value 498[M+H] +
Embodiment 1
3-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] propionic acid (E1)
Figure G200880012733XD00932
With 3-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] ethyl propionate (D5) (240mg) is dissolved in MeOH (5ml), the NaOH aqueous solution (5ml) with 2M is processed, and in stirred overnight at room temperature, then vacuum removes MeOH.Then reaction mixture with the NaOH aqueous solution dilution of 1M, with EtOAc (x2) washing, and is acidified to pH=3.This solution extracts with EtOAc (x3), and the extraction liquid salt water washing that merges, and then evaporation obtains crude product.It uses MDAP equipment purifying, obtains title compound (13mg), is white solid.δ H (CDCl 3, 400MHz): 1.44 (6H, d), 3.03 (2H, t), 4.68-4.75 (3H, m), 7.05 (1H, d), 7.60 (1H, d), 8.06 (1H, d), 8.11 (1H, s), (8.15 1H, d), 8.24 (1H, s), 8.56 (1H, s) .MS (ES): C 21H 19 35ClN 4O 4Calculated value 426; Measured value 427 (MH +).
Embodiment 1 (another kind of method)
3-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] propionic acid (E1)
Figure G200880012733XD00941
With 3-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] ethyl propionate (D5) (1.95g, 4.29mmol) is dissolved in ethanol.The NaOH (2.143ml, 4.29mmol) that adds 2M, and solution was placed 3 hours in room temperature.Ethanol evaporation, and mixture is dissolved in EtOAc and water, and use the acetic acid acidifying.With the dry (MgSO of organic layer 4) and evaporate the white solid that obtains (1.34g).δ H (d 6-DMSO, 400MHz): 1.37 (6H, d), 2.89 (2H, t), 4.65 (2H, t), 4.89 (1H, m), 7.45 (1H, d), 7.89 (1H, d), 8.06 (1H, d), 8.12 (1H, d), 8.20 (1H, s), 8.27 (1H, s), 8.55 (1H, s) .MS (ES): C 21H 19 35ClN 4O 4Calculated value 426; Measured value 427 (MH +).
Embodiment 2
3-(5-{5-[3-chloro-4-(oxyethyl group) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) Sodium Propionate (E2)
With 3-(5-{5-[3-chloro-4-(oxyethyl group) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) ethyl propionate (D57) (77mg, 0.17mmol) is suspended in the NaOH aqueous solution (2ml) and ethanol (2ml) of 2M.Mixture is in 50 ℃ of heating until dissolving is then cooling.Remove ethanol in vacuum, and filter out the gained solid, use ethanol, water and ether washing, then drying obtains title compound (63.6mg) under high vacuum, is pink solid.δ H (methyl alcohol-d 4, 400MHz) 1.49 (3H, t), 2.77 (2H, t), 4.25 (2H, q), 4.71 (2H, t), 7.27 (1H, d), 7.82 (1H, d), 8.09-8.25 (4H, m), 8.55 (1H, s) .MS (ES): C 20H 17 35ClN 4O 4Calculated value; 412 measured value (MH +) 413.
Embodiment 3
3-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] Sodium Propionate (E3)
Figure G200880012733XD00951
With 3-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] ethyl propionate (D58) (99mg, 0.22mmol) be dissolved in ethanol (2ml), and add the NaOH aqueous solution (0.55ml) of 2M.React on 80 ℃ of heating 30 minutes, and described agent dissolves.Reaction mixture is cooled to room temperature, then places and spend the night.Filter out the gained precipitation, wash with water, then with the ether washing, obtain title compound (29.1mg) by vacuum and heating drying.δ H (methyl alcohol-d 4, 400MHz) 1.46 (6H, d), 2.78, (2H, t), 4.71 (2H, t), 4.88-5.00 (the estimation 1H[weak peak that overlaps], m) .7.43 (1H, d), (7.83 1H, d), 8.10-8.17 (2H, m), 8.37-8.49 (2H, m), (8.56 1H, s), MS (ES): C 22H 19N 5O 4Calculated value 417; Measured value (MH +) 418.
Embodiment 4
3-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] propionic acid (E4)
Figure G200880012733XD00961
By in 50 ℃ of heating, with 3-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] ethyl propionate (D58) (523mg, 1.17mmol) is dissolved in the mixture of ethanol (20ml) and THF (10ml).Be cooled to room temperature and add the NaOH (1ml, 2mmol) of 2N, then placed 30 minutes in room temperature.Steaming desolventizes, and adds entry (50ml), and washs with EtOAc (50ml).The acidifying water layer is also used EtOAc (50ml) extraction product.With the dry (MgSO of EtOAc layer 4), and evaporating solvent.Grind with ether and obtain title compound (139mg), be beige solid.δ H (d 6-DMSO, 400MHz): 1.39 (6H, d), 2.90 (2H, t), 4.66 (2H, t), 4.94-5.03 (1H, m), 7.57 (1H, d), 7.90 (1H, d), 8.07 (1H, dd), 8.28 (1H, d), 8.42 (1H, dd), 8.54 (2H, dd), 12.35 (1H, broad s) .MS (ES): C 22H 19N 5O 4Calculated value 417; Measured value (MH +) 418.
Embodiment 5
4-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] butyric acid (E5)
Figure G200880012733XD00971
With 4-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-2H-indazole-2-yl] ethyl butyrate (D44) (73mg, 0.16mmol), 33% dimethyl amine heated 1 hour in 160 ℃ in microwave in ethanol (4ml).Add 2 to drip, then heated 3 hours in 160 ℃ in microwave.Make the reaction cool overnight, then be evaporated in a vacuum dried.Carry out (attempt) MDAP purifying, obtain thick material.
To 4-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-2H-indazole-2-yl] add the NaOH aqueous solution (1ml) of 2M in the solution of ethyl butyrate (D44) (65mg, 0.14mmol) in ethanol (10ml).In 80 ℃ of heating 1 hour, then place in room temperature and spend the night.Evaporating solvent, and thick material is by the MDAP purifying.The gained material grinds with ether and water and obtains thick material.
Merge the thick material of each batch and pass through the MDAP purifying, the fraction freeze-drying that will contain product obtains title compound (11.1mg).δ H (d 6-DMSO, 400MHz) 1.39 (6H, d), 2.01-2.11 (2H, m), (2.23 2H, t), 4.50 (2H, t), (4.98 1H, septet), 7.56 (1H, d), (7.86 1H, d), 8.07 (1H, dd), (8.28 1H, d), 8.42 (1H, dd), (8.52 1H, d), 8.57 (1H, dd) .MS (ES): C 23H 21N 5O 4Calculated value 431; Measured value (MH +) 432.
Embodiment 6
3-(5-{5-[4-[(1-methylethyl) oxygen base]-3-(trifluoromethyl) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) propionic acid (E6)
Figure G200880012733XD00972
With 3-(5-{5-[4-[(1-methylethyl) oxygen base]-3-(trifluoromethyl) phenyl]-1,2,4-oxadiazole-3-yl }-1H-indazole-1-yl) ethyl propionate (D59) (82mg, 0.17mmol) be dissolved in ethanol (4ml), then add the NaOH aqueous solution (0.5ml) of 2M.React on stirred overnight at room temperature, then ethanol evaporation in a vacuum.Add ethyl acetate, but compound exists all in water layer and organic layer, and mixture is evaporated.By MDAP (with 2 crowdes) purifying, then freeze-drying obtains title compound (28.7mg).δ H (400MHz, d 6-DMSO) 1.36 (6H, d), 2.88 (2H, t), (4.65 2H, t), 4.98 (1H, septets .), (7.59 1H, d), 7.89 (1H, d), (8.07 1H, dd), 8.27 (1H, d), (8.32 1H, d), 8.41 (1H, dd), 8.57 (1H, dd) .MS (ES): C 22H 19F 3N 4O 4Calculated value 460; Measured value (MH +) 461.
Embodiment 7
4-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] butyric acid (E7)
Figure G200880012733XD00981
With 2M sodium hydroxide (1ml, 2mmol) join 4-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] (N2666-13-A1) (0.062g of ethyl butyrate (D60), 0.132mmol) in solution in ethanol (5ml), and reaction is heated to 80 ℃ with dissolving, then cooling 2 hours.Evaporation is dissolved in ethyl acetate/water (each 25ml), and uses the acetic acid acidifying.With organic layer dry (sal epsom), evaporation with the 15ml methylbenzene azeotropic, and is ground with ether.Filtration obtains title compound (42mg), is white solid.δ H (d 6-DMSO, 400MHz): 1.37 (6H, d), 2.09 (2H, m), (2.24 2H, t), 4.54 (2H, t), (4.90 1H, m), 7.47 (1H, d), (7.62 1H, dd), 7.97 (1H, d), (8.01 1H, d), 8.17 (1H, dd), (8.26 1H, d), 8.56 (1H, s) .MS (ES): C 22H 21 35ClN 4O 4Calculated value 440; Measured value (MH +) 441.
Embodiment 7 (another kind of method)
4-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] butyric acid (E7)
Figure G200880012733XD00991
With 4-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] ethyl butyrate (D60) (1.5g, 3.20mmol) solution in hot ethanol (15ml) is with sodium hydroxide (7.5ml, 15.00mmol) process, and placed 1 hour.Evaporating solvent, and resistates is allocated in ethyl acetate/water.Add acetic acid to regulate pH to 5, separate each layer.Organic layer is passed through dried over mgso, and evaporation.Thick material grinds with ether and obtains title compound (1.1g), is white solid.δ H (d 6-DMSO, 400MHz): 1.37 (6H, d), 2.08 (2H, m), (2.23 2H, t), 4.54 (2H, t), (4.90 1H, m), 7.47 (1H, d), (7.62 1H, dd), 7.97 (1H, d), (8.01 1H, d), 8.17 (1H, dd), (8.26 1H, d), 8.56 (1H, s) .MS (ES): C 22H 21 35ClN 4O 4Calculated value 440; Measured value (MH +) 441.
Embodiment 8
3-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] propionic acid (E8)
Figure G200880012733XD00992
With 3-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] ethyl propionate (D61) (380mg, 0.835mmol) be dissolved in hot ethanol (4ml), add 2M sodium hydroxide (2ml, 4.00mmol), and stirred solution 30 minutes.Form precipitation.Ethanol evaporation, and resistates is allocated in EtOAc/ water.Water layer neutralizes with AcOH, and separates each layer.Organic layer is through dried over mgso, and evaporation.Thick material grinds with ether and obtains (240mg) white solid.δ H (d 6-DMSO) 1.36 (6H, d), 2.90 (2H, t), (4.68 2H, t), 4.89 (1H, m), (7.46 1H, d), 7.59 (1H, t), (7.99 2H, d), 8.16 (1H, dd), (8.25 1H, d), 8.55 (1H, s), 11.90 (1H, b r.s) .MS (ES): C 21H 19 35ClN 4O 4Calculated value 426; Measured value (MH +) 427.
Following embodiment is by similar hydrolysis reaction preparation.Except as otherwise noted, reaction is carried out between room temperature to 80 ℃.Reaction solvent is the mixture of the NaOH (aqueous solution) of ethanol and 2M.Sometimes add other solvent, as DCM, to help dissolving.In some cases, use other solvent in post-processing step, replace EtOAc as DCM.In some cases, reaction mixture acidifying (use acetic acid or hydrochloric acid) obtaining carboxylic acid, and is separated product in other situations with sodium salt.Evaporate in some cases some reaction solvents and cause the product precipitation, and this material passes through filtering separation.Perhaps, sometimes separating carboxylic acids and/or carboxylic acid esters then by being converted into sodium salt with sodium-hydroxide treatment.At E23, in the situation of E24, use hydrochloric acid acidizing reaction to obtain hydrochloride in aftertreatment.This compound is by MDAP or grind purifying.Sometimes use freeze-drying to obtain solid matter.
Figure G200880012733XD01011
Figure G200880012733XD01021
Figure G200880012733XD01031
Figure G200880012733XD01041
Figure G200880012733XD01051
Figure G200880012733XD01061
Figure G200880012733XD01081
Embodiment 42
4-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] butyric acid (E42)
Figure G200880012733XD01091
With 4-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl] ethyl butyrate (D63) (127mg, 0.27mmol) is dissolved in ethanol (5ml), then adds sodium hydroxide (11mg).Reaction mixture in microwave reactor in 100 ℃ the heating 1 hour, then cooling.The gained solid by filtration is separated, and then uses washing with acetone.By the MDAP purifying, then freeze-drying gained material obtains title compound (38.1mg).MS (ES): C 22H 21 35ClN 4O 4Calculated value 440; Measured value (MH +) 441.
The following example uses and above-mentioned similar method preparation.
Figure G200880012733XD01092
Embodiment 44
4-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-2H-indazole-2-yl] butyric acid (E44)
Figure G200880012733XD01101
With 4-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-2H-indazole-2-yl] ethyl butyrate (D45) (60mg, 0.13mmol) join in the solution of dimethyl amine in ethanol (4ml), then add several dripping.Mixture is heated to 160 ℃ in microwave reactor, kept 4 hours, and is then cooling.Remove in a vacuum solvent, and slightly material by the MDAP purifying, obtains title compound (28.3mg), is white solid.MS (ES): C 23H 21N 5O 4Calculated value 431; Measured value (MH +) 432.
Embodiment 45
3-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-PA (E45)
Figure G200880012733XD01102
In room temperature, to 3-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-2, add the NaOH (H of 2N in the solution of 2-ethyl dimethyl (D100) (85mg, 176mmol) in THF (2ml) 2O solution, 440 μ l, 880mmol), and the gained mixture spends the night in 50 ℃ of stirrings.Add the NaOH (H of 2N 2O solution, 440 μ l, 880mmol) and THF (2ml), and the gained mixture stirred for 2 nights in 80 ℃, then was cooled to room temperature.Remove most of THF in vacuum, and with the gained aqueous phase as acidified to pH 3.Leach the precipitation of formation, and use H 2The O washing, then drying obtains title compound (73mg, 91%) under 60 ℃ of vacuum, is white solid.MS (ES): C 23H 23ClN 4O 4Calculated value 454; Measured value (MH +) 455.
Following embodiment uses and the described similar method of embodiment 45 (E45), uses EtOH rather than THF to prepare as organic solvent:
Figure G200880012733XD01111
Embodiment 48
3-[4-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-PA (E48)
Figure G200880012733XD01112
With 3-[4-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-PA ethyl ester (D105) (50mg, 0.103mmol) is dissolved in ethanol (10ml).Add the sodium hydroxide (H of 2N 2O solution, 0.258ml, 0.517mmol), and the gained mixture spends the night in 60 ℃ of stirrings, then is cooled to room temperature.Evaporating solvent, then residue diluted with water.Then water extracts with EtOAc with the HCl aqueous solution neutralization of 2N.Organic phase H 2The O washing is through MgSO 4Drying, and vacuum concentration obtains title compound (22mg, 42%), is pale solid.MS (ES): C 22H 22ClN 5O 4Calculated value 455; Measured value (MH +) 456.
Embodiment 49
3-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-PA (E49)
Figure G200880012733XD01121
With 3-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-PA ethyl ester (D107) (50mg, 0.106mmol) is dissolved in ethanol (10ml).Add the sodium hydroxide (H of 2N 2O solution, 0.264ml, 0.528mmol), and the gained mixture was in stirring at room 2 hours.Remove in a vacuum most of solvent, and residue diluted with water.Then water extracts with EtOAc with the HCl aqueous solution neutralization of 2N.Organic phase H 2The O washing is through MgSO 4Dry and vacuum concentration obtains title compound (15mg, 30%), is light yellow solid.MS (ES): C 23H 24N 5O 4Calculated value 445; Measured value (MH +) 446.
Embodiment 50
4-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-2,2-acid dimethyl (E50)
Figure G200880012733XD01131
Use the method similar to Preparation Example 49 (E49), from 4-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-2,2-dimethyl butyrate acetoacetic ester (D108) obtains title compound (E50) (16mg, 32%).MS (ES): C 25H 25N 5O 4Calculated value 459; Measured value (MH +) 460.
Embodiment 51
3-[4-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-PA (E51)
Figure G200880012733XD01132
With 3-[4-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-2,2-ethyl dimethyl (D109) (100mg, 0.21mmol) solution in ethanol (2ml) is with 2M aqueous sodium hydroxide solution (1ml, excessive in a large number) process, then in 80 ℃ of heating 2 hours.Reaction mixture is cooling, and remove in a vacuum most of ethanol.Resistates water (3ml) dilution, and with dense HCl acidified aqueous solution, and extract with EtOAc (3x10ml).Dry and the vacuum concentration with the extraction liquid that merges.Resistates obtains required product (5mg, 5%) by the MDAP purifying, is colorless solid.MS (ES) C 24H 23N 5O 4Calculated value 445; Measured value (MH +) 446.
Embodiment 52
3-[5-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-PA (E52)
3-[5-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-oxadiazole-3-yl)-1H-indazole-1-yl]-2,2-ethyl dimethyl (D111) (100mg, 0.21mmol) solution in ethanol (2ml) processes with 2M aqueous sodium hydroxide solution (1ml), then under agitation heated 4 hours in 80 ℃.Reaction mixture is cooled to room temperature, and water (5ml) dilution.Aqueous solution Glacial acetic acid acidifying, and extract with EtOAc (2x10ml).The organic phase that merges is through MgSO 4Drying, and vacuum concentration.Resistates obtains required product (3mg, 3%) by the MDAP purifying, is colorless solid.MS (ES): C 22H 22ClN 5O 4Calculated value 455; Measured value (MH +) 456.
Following embodiment uses the described similar method preparation to embodiment 48 (E48).
Figure G200880012733XD01142
Figure G200880012733XD01151
S1P1 GTP γ S is in conjunction with test
The rat basophilic leukemia cell (RBL) of stably express S1P1 acceptor grows to 80% and merges, then collects in 10ml phosphate-buffered saline (PBS), and in 1200rpm centrifugal 5 minutes.After removing supernatant liquor, precipitation is hanged also homogenate again at test damping fluid (20mM HEPES pH 7.4,100mM NaCl, the 10mM MgCl of 20 volumes 2.6H 2O, 10 μ M GDP Saponin 10 μ g/ml) in.Centrifugal film suspension 20 minutes again under 20,000rpm, then homogenizing and centrifugal (spun) again.After centrifugal for the second time, throw out be resuspended in suitable volumes (every bottle of cell 1ml), and measure protein concn.
The concentrated reserve of S1P is carried out supersound process, then with initial concentration 10 -5M prepares a series of diluents.With the S1P of different concns and 0.3nM in 96 deep-well plates 35S-GTP γ S (NEN; Specific activity 1250Ci/mmol) film after incubation dilution (10 μ g/ hole).Carry out in conjunction with 45 minutes at 30 ℃, and stop combination by with Packard Universal collector, film being collected on the GF/B screen plate.Dry this plate is after 45 minutes, the Microscint 0 of 50 μ l added to respectively in each hole and measure in conjunction with situation Topcount NXT (Perkin Elmer) is upper.Use Graphpad Prism 4 analytical data and be expressed as stimulation percentage ratio (percentage stimulation above basal) over benchmark.The EC50 value is defined as the concentration of the 50% required agonist that produces maximal stimulation.
This GTP γ S in conjunction with the test in, the pEC50 of embodiments of the invention 1＞6.
S1P1 GTP γ S is in conjunction with test (another kind of method)
The step of all film preparations is all carried out under 4 ℃.The rat basophilic leukemia cell (RBL) of the rat hepatoma cell of stably express people S1P1 acceptor or stably express people S1P3 acceptor grows to 80% and merges, then be collected in the phosphate-buffered saline (PBS) of 10ml, and under 1200rpm centrifugal 5 minutes.After removing supernatant liquor, with the throw out resuspension, and in the glass Waring blender at damping fluid (the 50mM HEPES of 200ml, the 1mM leupeptin, 25 μ g/ml bacitracins, 1mM EDTA, 1mM PMS F, 2 μ M pepstatin A) in by two 15 seconds break (burst) with cell homogenization.After breaking for the first time, blendor was immersed in ice 5 minutes, and immerse ice after final breaking in 10～40 minutes, so that foam dissipates.Then the gained material was rotated 20 minutes under 500g, supernatant liquor was rotated 36 minutes under 48000g.But throw out is resuspended in above-mentioned the same do not contain in the damping fluid of PMSF and pepstatin A.Then force the gained material to pass the syringe needle of 0.6mm, obtain volume required (be generally initial cell throw out volume 4 times), aliquots containig and refrigerated storage are in-80 ℃.
People S1P1 rat liver cancer film (1.5 μ g/ hole) is adhered to measure damping fluid (HEPES 20mM, MgCl 210mM, NaCl 100mM, using KOH 5M to regulate pH is 7.4, also adds GDP 10 μ M FAC (finally measuring concentration) and saponin(e 90 μ g/ml FAC) in the approaching mensuration of flicker (SPA) pearl of wheat germ agglutinin (WGA) coating on (0.125mg/ hole).
In pre-coupling on ice after 30 minutes, pearl and film suspension are dispersed in white Greiner polypropylene LV384 orifice plate (5 μ l/ hole), wherein contain the compound of 0.1 μ 1.Then will measure the 5 μ g/ holes of making in buffered soln [ 35S]-GTP γ S (the final radioligand concentration of 0.5nM) adds on the agonist plate.Then centrifugal final mensuration mixture (cocktail) (10.1 μ l) 5 minutes under 1000rpm, read on the Viewlux reader immediately.
The concentration of all test compounds with 10mM is dissolved in DMSO, and uses 1 to prepare in 100%DMSO in 4 (1in4) dilution step, obtain the dose response curve of 11.Diluent is transferred on assay plate, guarantees that it is constant that DMSO concentration is determined on whole plate for all.
All data all are standardized as on each plate the mean value of 16 high and 16 low control wells.Then use four parameter curves.
In this test, the pEC50 of embodiment of the present invention 1-52＞5.
S1P3 GTP γ S is in conjunction with test
To adhere to and measure damping fluid (HEPES 20mM, MgCl from the S1P3 film of rat basophilic leukemia cell (RBL-2H3) (1.5 μ g/ hole) 23mM, NaCl 100mM, using KOH 5M to regulate pH is 7.4, also adds GDP 10 μ M FAC and saponin(e 90 μ g/ml FAC) in the SPA pearl that applies of WGA on (0.125mg/ hole).
In pre-coupling on ice after 30 minutes, pearl and film suspension are dispersed in white Greiner polypropylene LV384 orifice plate (5 μ l/ hole), wherein contain the compound of 0.1 μ l.Then will measure the 5 μ g/ holes of making in buffered soln [ 35S]-GTP γ S (the final radioligand concentration of 0.5nM) adds on the agonist plate.Then centrifugal final mensuration mixture (cocktail) (10.1 μ l) 5 minutes under 1000rpm, read on the Viewlux reader immediately.
The concentration of all test compounds with 10mM is dissolved in DMSO, and uses 1 to prepare with 100%DMSO in 4 dilution step, obtain the dose response curve of 11.Diluent is transferred on assay plate, guarantees that it is constant that DMSO concentration is determined on whole plate for all.
All data all are standardized as on each plate the mean value of 16 high and 16 low control wells.Then use four parameter curves.
In this test, embodiment 1-46 and 52 pEC50＜6.5, the pEC50 of many detected embodiment＜5.
Yeast is in conjunction with test
Produce yeast (yeast saccharomyces cerevisiae (Saccharomyces the cerevisiae)) cell of expressing people S1P1 acceptor by the ura3 chromogene seat that expression cassette (cassette) is integrated into yeast strain MMY23.This box comprises the DNA sequence dna of encoding human S1P1 acceptor, S1P1 5 ' end flank be yeast GPD promotor, and S1P1 3 ' end flank be yeast Transcription Termination subsequence.MMY23 expresses yeast/Mammals mosaic G-protein alpha subunit, and wherein 5 amino acid of the C-end of Gpa1 are by 5 amino acid replacements of the C-end of people G μ i1/2 (as (2000) such as Brown, described in Yeast 16:11-22).At 30 ℃, cell is growth (Guthrie and Fink (1991) in there is no fully synthetic (SC) (liquid Synthetic Complete (SC)) the yeast culture base of uridylic, tryptophane, VITAMIN B4 and leucic liquid, Methods in Enzymology, Vol.194) to logarithmic phase later stage (about 6OD 600/ ml).
Agonist is prepared into the DMSO solution of 10mM.Use 4 times of diluents (BiomekFX, Beckman) mensuration EC in DMSO 50Value (producing 50% desired concn of peak response).The DMSO solution (1% final mensuration volume) of agonist is transferred in black titer plate (Greiner, 384 holes).With cell with density 0.2OD 600/ ml is suspended in does not have Histidine, uridylic, tryptophane, VITAMIN B4 and leucine and replenish in the SC substratum of 0.1mM 3-aminotriazole, 0.1M sodium phosphate (pH 7.0) and 10 μ M fluorescein two-β-D-glucopyranosides (FDGlu).This mixture (50 μ l/ hole) is made an addition to assay plate, and (Multidrop 384, in the agonist in Labsystems).After 24 hours, use fluorescence titer plate reader (Tecan Spectrofluor or LJL Analyst, excitation wavelength: 485nm 30 ℃ of cultivations; Emission wavelength: 535nm) mensuration is degraded to because exoglucanase makes FDGlu the fluorescence that fluorescein generates, and described exoglucanase is the endogenous yeast enzyme that produces during the Growth of Cells that agonist stimulates.Fluorescence is mapped to compound concentration, and come matched curve repeatedly with four parameter fittings, to obtain the mass action value.According to following Equation for Calculating effect (E max):
E max=Max [compounds X]-Min [compounds X]/ Max [S1P]-Min [S1P]X100%
Wherein, Max [compounds X]And Min [compounds X]Be respectively match maximum value and minimum value that the mass action curve by compounds X obtains, Max [S1P]And Min [S1P]Be match maximum value and the minimum value that is obtained by S1P (can be obtained by Sigma) mass action curve respectively.Mol ratio (EMR) value according to following Equation for Calculating equivalence:
EMR=EC 50[compounds X]/ EC 50[S1P]
EC wherein 50[compounds X]The EC of compounds X 50, EC 50[S1P]The EC of S1P 50
In this test, the pEC50 of the exemplary compounds of the present invention of test＞5.

Claims (20)

1. formula (I) compound or its salt:
Figure FSB00001003763200011
Wherein
R 5And R 6In one be hydrogen, another is (a)
Figure FSB00001003763200012
R 3And R 4One of be (b)
Figure FSB00001003763200013
A is thienyl, pyridyl or phenyl; And
R 1Be two substituting groups, described substituting group is independently selected from: chlorine, bromine, isopropoxy, propoxy-, methoxyl group, 1-methyl propoxy-, cyano group, trifluoromethyl, trifluoromethoxy, cyclohexyl, piperidyl, pyrrolidyl, ethyl, 2-methyl-propyl, phenyl and cyclopentyloxy; And
R 2Be hydrogen; And
R 7Be hydrogen; And
Z is ethylidene or propylidene, is optionally separately replaced together with-dimethyl.
2. according to claim 1 formula (I) compound or its salt, wherein
R 6Be (a), and R 5Be hydrogen; And
A is thienyl, pyridyl or phenyl; And
R 1Be two substituting groups, described substituting group is independently selected from: chlorine, bromine, isopropoxy, propoxy-, methoxyl group, 1-methyl propoxy-, cyano group, trifluoromethyl, trifluoromethoxy, cyclohexyl, piperidyl, pyrrolidyl, ethyl, 2-methyl-propyl, phenyl and cyclopentyloxy; And
R 2Be hydrogen; And
R 7Be hydrogen; And
Z is ethylidene or propylidene, is optionally separately replaced together with-dimethyl.
3. according to claim 1 formula (I) compound or its salt, wherein
R 5Be (a), and R 6Be hydrogen; And
A is thienyl, pyridyl or phenyl; And
R 1Be two substituting groups, described substituting group is independently selected from: chlorine, bromine, isopropoxy, propoxy-, methoxyl group, 1-methyl propoxy-, cyano group, trifluoromethyl, trifluoromethoxy, cyclohexyl, piperidyl, pyrrolidyl, ethyl, 2-methyl-propyl, phenyl and cyclopentyloxy; And
R 2Be hydrogen; And
R 7Be hydrogen; And
Z is ethylidene or propylidene, is optionally separately replaced together with-dimethyl.
4. formula (IA) compound or its salt:
Figure FSB00001003763200021
R wherein 2, R 5, R 6, R 7Define according to claim 1 with Z.
5. according to claim 4 formula (IA) compound or its salt, wherein
R 6Be (a), and R 5Be hydrogen; And
A is phenyl; And
R 1Be two substituting groups, described substituting group is independently selected from: chlorine, isopropoxy, cyano group, trifluoromethyl, trifluoromethoxy, piperidyl, ethyl and phenyl; And
R 2Be hydrogen; And
R 7Be hydrogen; And
Z is ethylidene or propylidene.
6. according to claim 4 formula (IA) compound or its salt, wherein
R 6Be (a), and R 5Be hydrogen; And
A is phenyl; And
R 1Be chlorine and isopropoxy; And
R 2Be hydrogen; And
R 7Be hydrogen; And
Z is ethylidene or propylidene.
7. according to claim 4 formula (IA) compound or its salt, wherein
R 5Be (a), and R 6Be hydrogen; And
A is phenyl; And
R 1Be chlorine and isopropoxy; And
R 2Be hydrogen; And
R 7Be hydrogen; And
Z is ethylidene.
8. formula (IB) compound or its salt:
Figure FSB00001003763200031
R wherein 2, R 5, R 6, R 7Define according to claim 1 with Z.
9. according to claim 8 formula (IB) compound or its salt, wherein
R 6Be (a), and R 5Be hydrogen; And
A is phenyl; And
R 1Be two substituting groups, described substituting group is independently selected from: chlorine, isopropoxy and cyano group; And
R 2Be hydrogen; And
R 7Be hydrogen; And
Z is ethylidene or propylidene.
10. according to claim 8 formula (IB) compound or its salt, wherein
R 5Be (a), and R 6Be hydrogen; With
A is phenyl; And
R 1Be two substituting groups, described substituting group is independently selected from: chlorine, isopropoxy, phenyl and trifluoromethyl; And
R 2Be hydrogen; And
R 7Be hydrogen; And
Z is propylidene.
11. compound is selected from:
3-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-
Figure FSB00001003763200041
Diazole-3-yl)-1H-indazole-1-yl] propionic acid
3-(5-{5-[3-chloro-4-(oxyethyl group) phenyl]-1,2,4-
Figure FSB00001003763200042
Diazole-3-yl }-1H-indazole-1-yl) propionic acid
3-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4- Diazole-3-yl)-1H-indazole-1-yl] propionic acid
4-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-
Figure FSB00001003763200044
Diazole-3-yl)-1H-indazole-1-yl] butyric acid
3-(5-{5-[4-[(1-methylethyl) oxygen base]-3-(trifluoromethyl) phenyl]-1,2,4-
Figure FSB00001003763200045
Diazole-3-yl }-1H-indazole-1-yl) propionic acid
4-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-
Figure FSB00001003763200046
Diazole-3-yl)-1H-indazole-1-yl] butyric acid
3-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-
Figure FSB00001003763200047
Diazole-3-yl)-1H-indazole-1-yl] propionic acid
3-(5-{5-[3-chloro-4-(propoxy-) phenyl]-1,2,4-
Figure FSB00001003763200048
Diazole-3-yl }-1H-indazole-1-yl) propionic acid
3-[5-(5-{3-chloro-4-[(trifluoromethyl) oxygen base] phenyl }-1,2,4-
Figure FSB00001003763200049
Diazole-3-yl)-1H-indazole-1-yl] propionic acid
3-(5-{5-[4-cyclohexyl-3-(trifluoromethyl) phenyl]-1,2,4-
Figure FSB000010037632000410
Diazole-3-yl }-1H-indazole-1-yl) propionic acid
3-(5-{5-[4-(methoxyl group)-3-(trifluoromethyl) phenyl]-1,2,4-
Figure FSB000010037632000411
Diazole-3-yl }-1H-indazole-1-yl) propionic acid
[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4- Diazole-3-yl)-1H-indazole-1-yl] acetic acid
3-(5-{5-[3-chloro-4-(methoxyl group) phenyl]-1,2,4-
Figure FSB000010037632000413
Diazole-3-yl }-1H-indazole-1-yl) propionic acid
3-(5-{5-[3-ethyl-4-(piperidino) phenyl]-1,2,4- Diazole-3-yl }-1H-indazole-1-yl) propionic acid
3-{5-[5-(4-cyclohexyl-3-ethylphenyl)-1,2,4-
Figure FSB000010037632000415
Diazole-3-yl]-1H-indazole-1-yl } propionic acid
3-(5-{5-[3-cyano group-4-(2-methyl-propyl) phenyl]-1,2,4-
Figure FSB000010037632000416
Diazole-3-yl }-1H-indazole-1-yl) propionic acid
3-[5-(5-{3-bromo-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-
Figure FSB000010037632000417
Diazole-3-yl)-1H-indazole-1-yl] propionic acid
3-(5-{5-[3-chloro-4-(2-methyl-propyl) phenyl]-1,2,4-
Figure FSB00001003763200051
Diazole-3-yl }-1H-indazole-1-yl) propionic acid
4-(5-{5-[4-[(1-methylethyl) oxygen base]-3-(trifluoromethyl) phenyl]-1,2,4-
Figure FSB00001003763200052
Diazole-3-yl }-1H-indazole-1-yl) butyric acid
3-{5-[5-(2-cyano group-4-xenyl)-1,2,4- Diazole-3-yl]-1H-indazole-1-yl } propionic acid
3-(5-{5-[3-chloro-4-(1-pyrrolidyl) phenyl]-1,2,4- Diazole-3-yl }-1H-indazole-1-yl) propionic acid
4-[5-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-
Figure FSB00001003763200055
Diazole-3-yl)-1H-indazole-1-yl] the butyrates hydrochlorate
4-[5-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4- Diazole-3-yl)-2H-indazole-2-yl] the butyrates hydrochlorate
4-(5-{5-[2-(trifluoromethyl)-4-xenyl]-1,2,4- Diazole-3-yl }-1H-indazole-1-yl) butyric acid
4-(5-{5-[2-(trifluoromethyl)-4-xenyl]-1,2,4-
Figure FSB00001003763200058
Diazole-3-yl }-2H-indazole-2-yl) butyric acid
3-[4-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-
Figure FSB00001003763200059
Diazole-3-yl)-1H-indazole-1-yl] propionic acid
3-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4- Diazole-3-yl)-2H-indazole-2-yl] propionic acid
3-[4-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-
Figure FSB000010037632000511
Diazole-3-yl)-2H-indazole-2-yl] propionic acid
4-[4-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-
Figure FSB000010037632000512
Diazole-3-yl)-1H-indazole-1-yl] butyric acid
4-(4-{5-[3-ethyl-4-(piperidino) phenyl]-1,2,4-
Figure FSB000010037632000513
Diazole-3-yl }-1H-indazole-1-yl) butyric acid
4-(4-{5-[2-(trifluoromethyl)-4-xenyl]-1,2,4-
Figure FSB000010037632000514
Diazole-3-yl }-1H-indazole-1-yl) butyric acid
4-[4-(5-{3-chloro-4-[(trifluoromethyl) oxygen base] phenyl }-1,2,4-
Figure FSB000010037632000515
Diazole-3-yl)-1H-indazole-1-yl] butyric acid
3-(4-{5-[4-[(1-methylethyl) oxygen base]-3-(trifluoromethyl) phenyl]-1,2,4- Diazole-3-yl }-1H-indazole-1-yl) propionic acid
4-[4-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-
Figure FSB000010037632000517
Diazole-3-yl)-1H-indazole-1-yl] butyric acid
4-(4-{5-[4-phenyl-5-(trifluoromethyl)-2-thienyl]-1,2,4-
Figure FSB000010037632000518
Diazole-3-yl }-1H-indazole-1-yl) butyric acid
4-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-
Figure FSB00001003763200061
Diazole-3-yl)-2H-indazole-2-yl] butyric acid
3-{5-[5-(3-cyano group-4-{[(1S)-1-methyl-propyl] the oxygen base } phenyl)-1,2,4-
Figure FSB00001003763200062
Diazole-3-yl]-1H-indazole-1-yl } propionic acid sodium salt
3-{5-[5-(3-cyano group-4-{[(1R)-1-methyl-propyl] the oxygen base } phenyl)-1,2,4-
Figure FSB00001003763200063
Diazole-3-yl]-1H-indazole-1-yl } propionic acid sodium salt
3-(5-{5-[3-cyano group-4-(cyclopentyloxy) phenyl]-1,2,4-
Figure FSB00001003763200064
Diazole-3-yl }-1H-indazole-1-yl) propionic acid
4-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4- Diazole-3-yl)-1H-indazole-1-yl] butyric acid
4-(5-{5-[4-phenyl-5-(trifluoromethyl)-2-thienyl]-1,2,4-
Figure FSB00001003763200066
Diazole-3-yl }-1H-indazole-1-yl) butyric acid
4-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-
Figure FSB00001003763200067
Diazole-3-yl)-2H-indazole-2-yl] butyric acid
3-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-
Figure FSB00001003763200068
Diazole-3-yl)-1H-indazole-1-yl]-PA
3-[5-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4- Diazole-3-yl)-2H-indazole-2-yl]-PA
3-[4-(5-{3-chloro-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4- Diazole-3-yl)-1H-indazole-1-yl]-PA
3-[4-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-
Figure FSB000010037632000611
Diazole-3-yl)-1H-indazole-1-yl]-PA
3-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-
Figure FSB000010037632000612
Diazole-3-yl)-1H-indazole-1-yl]-PA
4-[5-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-
Figure FSB000010037632000613
Diazole-3-yl)-1H-indazole-1-yl]-2, the 2-acid dimethyl
3-[4-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-
Figure FSB000010037632000614
Diazole-3-yl)-1H-indazole-1-yl]-PA
3-[5-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4-
Figure FSB000010037632000615
Diazole-3-yl)-1H-indazole-1-yl]-PA
4-[4-(5-{3-cyano group-4-[(1-methylethyl) oxygen base] phenyl }-1,2,4-
Figure FSB00001003763200071
Diazole-3-yl)-1H-indazole-1-yl]-2, the 2-acid dimethyl
4-[4-(5-{5-chloro-6-[(1-methylethyl) oxygen base]-the 3-pyridyl }-1,2,4- Diazole-3-yl)-1H-indazole-1-yl]-2, the 2-acid dimethyl
Or its salt.
12. according to claim 1-11, the compound of any one is for the preparation of the purposes in the medicine for the treatment of receptor-mediated illness or disease by S1P1.
13. purposes according to claim 12, wherein said illness or disease are multiple sclerosis, autoimmune disorder, chronic inflammatory illness, asthma, inflammatory neuropathy, sacroiliitis, transplanting, Crohn disease, lupus erythematosus, psoriasis, ischemia reperfusion injury, noumenal tumour and metastases, vascular disease, antalgesic, acute disease viral disease, inflammatory bowel, Regular Insulin and non-insulin-dependent diabetes mellitus (NIDDM).
14. purposes according to claim 12, wherein said illness or disease are ulcerative colitis.
15. according to claim 12 or 13 purposes, wherein said illness or disease are multiple sclerosis.
16. pharmaceutical composition comprises the compound of any one in claim 1-11.
17. the method for the pharmaceutical composition of preparation claim 16, described method comprises formula (I) compound of any one in claim 1-11 or its pharmacy acceptable salt and pharmaceutically acceptable carrier or mixed with excipients.
18. the method for preparation formula (IA) compound, as shown in following scheme:
Figure FSB00001003763200073
Scheme 1
Wherein, R 1, R 2, definition in A and Z such as claim 1,
Formula (III) compound is converted into formula (IV) compound by processing with hydrazine hydrate, and perhaps formula (IV) compound is commercially available;
Formula (IV) compound is processed with hydroxylamine hydrochloride and sodium bicarbonate in methyl alcohol and is converted into formula (V) compound by under the high temperature of 50 ℃;
Formula (V) compound is processed with the carboxylic acid of formula (VI) in DMF and is converted into formula (VII) compound by under EDClHCl and HOBt existence;
Formula (VII) compound is processed with alkylating agent (VIII) in DMF and is converted into formula (IX) compound by under existing at cesium carbonate;
Formula (IX) compound is converted into formula (IA) compound by processing with aqueous sodium hydroxide solution in methyl alcohol.
19. the method for preparation formula (IB) compound, wherein, R 1, R 2, definition in A and Z such as claim 1:
Formula (VII) and the reaction that (VIII) generates (IX) also produce formula (X) compound of isomery, as shown in following scheme; With formula (VIII) alkylating agent and alkaline purification production (IX) and (X) mixture of compound, they separate by chromatography formula (VII) compound in solvent;
Figure FSB00001003763200081
Formula (X) compound is converted into formula (IB) compound by the hydrolysing step described in following scheme
Figure FSB00001003763200082
20. method according to claim 19, wherein solvent is DMF, and alkali is cesium carbonate.
CN200880012733.XA 2007-04-19 2008-04-17 Oxadiazole substituted indazole derivatives for use as sphingosine 1-phosphate (s1p) agonists Expired - Fee Related CN101663297B (en)

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