US008158657B2

(12) United States Patent (10) Patent No.: US 8,158,657 B2 Tanaka et al. (45) Date of Patent: Apr. 17, 2012

(54) HETEROCYCLES SUBSTITUTED PYRIDINE 6,080,767 A 6/2000 Klein et al. DERVATIVES AND ANTIFUNGALAGENT 6,140,131 A 10/2000 Sunakawa et al. 6,174.905 B1 1/2001 Suzuki et al. CONTAINING THEREOF 6,200,975 B1 3/2001 Carling et al. 6,235,728 B1 5, 2001 Golik et al. (75) Inventors: Keigo Tanaka, Tsukuba (JP); Satoshi 6.255,318 B1 7/2001 Bedard et al. 6,310,203 B1 10/2001 Carling et al. Inoue, Tsukuba (JP); Norio Murai, 6,313,127 B1 1 1/2001 Waterson et al. Tsukuba (JP); Masayuki Matsukura, 6,319,944 B1 1 1/2001 Claiborne et al. Tsukuba (JP); Kazutaka Nakamoto, 6,340,690 B1 1/2002 Bachand et al. Tsukuba (JP); Shuji Shirotori, Tsukuba 6,380,218 B1 4/2002 Marfat et al. (JP); Shinya Abe, Tsukuba (JP) 6,407,116 B1 6/2002 Kajino et al. (Continued) (73) Assignee: Eisai R&D Management Co., Ltd., Tokyo (JP) FOREIGN PATENT DOCUMENTS DE 1972.71 17 A1 1, 1999 (*) Notice: Subject to any disclaimer, the term of this (Continued) patent is extended or adjusted under 35 U.S.C. 154(b) by 0 days. OTHER PUBLICATIONS Okawa et al., Synthesis, No. 10. pp. 1467-1475 (1998). (21) Appl. No.: 12/704,637 AKos Screening Library, Feb. 7, 2006. CAS Registry No. 434304 24-2. (22) Filed: Feb. 12, 2010 Aurora Screening Library, Jan. 1, 2007, CAS Registry No. (RN): 431922-54-2. (65) Prior Publication Data US 2010/O168173 A1 Jul. 1, 2010 (Continued) Primary Examiner — Patricia Morris Related U.S. Application Data (74) Attorney, Agent, or Firm — Birch, Stewart, Kolasch & (62) Division of application No. 1 1/589,128, filed on Oct. Birch, LLP 30, 2006, now Pat. No. 7,691,882. (57) ABSTRACT (60) Provisional application No. 60/731,267, filed on Oct. 31, 2005, provisional application No. 60/753,391, An object of the present invention is to provide an antifungal filed on Dec. 27, 2005. agent which has excellent antifungal effects and is Superior in terms of its physical properties, safety and metabolic stability. (30) Foreign Application Priority Data According to the present invention, there is disclosed a com pound represented by the following formula (I), or a salt Oct. 31, 2005 (JP) ...... 2005-31768O thereof: Dec. 27, 2005 (JP) ...... 2005-374.395 (51) Int. Cl. (I) A6 IK3I/443 (2006.01) N CO7D 413/04 (2006.01) (52) U.S. Cl...... 514/340; 546/272.1 is-N-C-O--N 71 (58) Field of Classification Search ...... 546/272.1; 2 514/340 RI N R2 See application file for complete search history. wherein R' represents a hydrogen atom, a halogen atom, an (56) References Cited amino group, a C- alkyl group, a Coalkoxy group or a C alkoxy C, alkyl group; R represents a hydrogen atom, a U.S. PATENT DOCUMENTS C- alkyl group, an amino group or a di C-alkylamino 4,576.956 A 3, 1986 Makisumi et al. 4,720.493 A 1, 1988 Kawakita et al. group; one of X and Y is a nitrogen atom while the other is a 4,785,010 A 11, 1988 Zoller et al. nitrogen atom or an oxygen atom; ring A represents a 5- or 5,034,393 A 7, 1991 Hackler et al. 6-member heteroaryl ring or a benzene ring which may have 5,070,082 A 12/1991 Murdocket al. a halogen atom, or 1 or 2 C- alkyl groups; Z represents a 5,208,247 A 5, 1993 Trova et al. single bond, a methylene group, an ethylene group, an oxygen 5,296.484 A 3/1994 Coghlan et al. 5,328,921 A 7, 1994 Trova et al. atom, a sulfur atom, —CH2O— —OCH , —NH-. 5,350,749 A 9, 1994 Hackler et al. CH-NH , -NHCH , —CHS , or -SCH. : R 5,371,086 A 12/1994 Takemoto et al. represents a hydrogen atom, a halogen atom, a C- alkyl 5,691,136 A 1 1/1997 Lupski et al. group, a C-8 cycloalkyl group, a Co-o aryl group, a 5- or 5,691,336 A 11/1997 Dornet al. 5,710, 171 A 1/1998 Dinsmore et al. 6-member heteroaryl group, or 5- or 6-member non-aromatic 5,747,518 A 5, 1998 Yoshikawa et al. heterocyclic group which may have 1 or 2 substituents; and 5,852,042 A 12/1998 Jakobi et al. R" represents a hydrogen atom or a halogen atom. 5,945,431 A 8, 1999 Jin et al. 6,022,884 A 2/2000 Mantlo et al. 14 Claims, No Drawings US 8,158,657 B2 Page 2

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No. Second Office Action issued Jan. 12, 2012, in Chinese Patent Appli 12/704,643. cation No. 200680040781.0, with English translation. US 8,158,657 B2 1. 2 HETEROCYCLES SUBSTITUTED PYRONE However, groups of the compounds disclosed in Patent DERVATIVES AND ANTIFUNGALAGENT Document 1 have 2-benzyl pyridine moieties as the common CONTAINING THEREOF structure, clearly differing structurally from compounds according to the present invention. In addition, the groups of CROSS-REFERENCE TO RELATED the compounds disclosed in Patent Document 1 bear the APPLICATIONS problem that, although these compounds demonstrate activi ties in vitro, they are easily metabolized inside the body. The This application is a 37 CFRS1.53(b) divisional applica group of compounds disclosed in Patent Document 2 exhibits tion which claims priority to U.S. application Ser. No. 1 1/589, excellent antifungal activity, but the group of representative 128 filed Oct. 30, 2006, now U.S. Pat. No. 7,691,882, which 10 compounds has the structure represented by the following claims priority to U.S. Provisional Application No. 60/731, formula: 267 filed Oct. 31, 2005, Japanese Patent Application No. 2005-317680 filed Oct. 31, 2005, U.S. Provisional Applica tion No. 60/753,391 filed Dec. 27, 2005, and Japanese Patent 15 Application No. 2005-374395 filed in Dec. 27, 2005, the (D-x-CH-O disclosures of which are hereby incorporated by reference in their entireties. A'=optionally substituted 3-pyridyl or quinolyl, etc. FIELD OF THE INVENTION X = C(=O) NH, NH CO-O) , etc. The present invention relates to heterocyclic substituted E=furyl, thienyl, pyrrolyl, phenyl, pyridyl, tetrazolyl, thia pyridine derivatives and to antifungal agents comprising the Zolyl, or pyrazolyl SaC. Looking only at those having pyridine ring skeletons, this 25 group differs structurally from the compounds according to DESCRIPTION OF THE RELATED ART the present invention in that the commonstructure has a single ring bound via an amidemethylene linker at the pyridine ring In recent years, managements of opportunistic infections 3-position. have become more and more significant more than ever Patent Documents 3 to 5 also provide examples of related because of an increase in the number of elderly people and 30 art with structures similar to the compounds according to the immunocompromised patients as a result of advanced che present invention. Patent Documents 3 and 4 describe pyri motherapies or the like. As demonstrated by the fact that dine derivatives substituted by a pyrazole ring, which are used opportunistic infections are occurring one after another by as glycine transporter inhibitors or 5-HT receptor ligands, different avirulent pathogen, it is shown that the problem of while Patent Document 5 describes 5-member heterocyclic infectious disease will not ends as long as there are underlying 35 substituted pyridine derivatives which are used as an AGE diseases that diminish the immune functions of patients. Con disruptor and inhibitor. sequently, new strategies for infectious diseases control, However, Patent Documents 3 to 5 do not disclose the including the problem of drug-resistant pathogen, will be one compounds according to the present invention, and the anti of the important issues in the Soon-to-come aged Society. fungal effects of the compounds disclosed in Patent Docu In the field of antifungal agents, heretofore, for instance, 40 ments 3 to 5 against Candida, Aspergillus, Cryptococcus and amphotericine B which is based on a polyene skeleton, flu the like which are common fungi in human fungal disease are conazole, itraconazole and Voriconazole which are based on not disclosed. an azole skeleton, or the like, have been developed for the treatment of deep seated mycoses. Most of pre-existing drugs Patent Document 1 International Publication WO 02/04626 already available commercially have similar mechanism of 45 pamphlet action, and currently, the appearance of azole-resistant fungi Patent Document 2 International Publication WO or the like has been problems. 05/033079 pamphlet In recent years, as a 1.3-3-glucan synthetase inhibitor with Patent Document 3 International Publication WO a novel mechanism, naturally occurring compound-derived 03/031435 pamphlet cyclic hexapeptides caspofungin and micafungin or the like, 50 have been developed; however, from the fact that these agents Patent Document 4 International Publication WO only exist in injectable form, they are not yet sufficient prac 04/089931 pamphlet tically as antifungal agents. Patent Document 5 International Publication WO Since there have been the situations that the pre-existing 02/085897 pamphlet antifungal agents are insufficient for treatment of the deep 55 seated mycoses, there is a demand and need for development BRIEF SUMMARY OF THE INVENTION of agents which are based on a novel mechanism and are of high safety. As the related art relevant to antifungal agents based on It is an object of the present invention to provide an anti Such a novel mechanism, Patent Documents 1 and 2 describe 60 fungal agent which has excellent antifungal action not found pyridine derivatives which demonstrates effects against the in the antifungal agents in the prior art, and which is also onset, progress, and persistence of infections by inhibiting the excellent in terms of property, safety and metabolic stability. expression of cell wall proteins, inhibiting the cell wall As a result of exhaustive research conducted in view of the assembly and also adhesion onto cells, and preventing patho above circumstances, the present inventors have Succeeded in gens from showing pathogenicity, with the process which 65 synthesizing novel pyridine derivatives (hereinafter, the com transports GPI (Glycosylphosphatidylinositol)-anchored pounds of the present invention) represented by the following proteins to the cell wall being inhibited. formula (I): US 8,158,657 B2 3 4 2 a compound represented by the following formula (I"), or a salt thereof: X (I) N. R N (O-ON 71 R3 (I) 2 R1 N R2 and having a chemical structure in which a pyridine ring and 10 a 5- or 6-member heteroaryl ring or benzene ring are joined with a 5-member heteroaryl methyl group as a linker, and wherein R' represents a hydrogen atom, a halogen atom, an have perfected the present invention upon discovering that amino group, a C- alkyl group, a Coalkoxy group or a C these compounds have excellent antifungal action. 15 alkoxy Calkyl group; That is, the present invention provides: R represents a hydrogen atom or an amino group; 1 a compound represented by the following formula (I), or one of X and Y is a nitrogen atom while the other is a a salt thereof: nitrogen atom or an oxygen atom; ring A represents a 5- or 6-member heteroaryl ring or a benzene ring; (I) Z represents a methylene group, an oxygen atom, N CHO , OCH , NH NHCH, O S-a-C-O--N 71 - CH-NH ; 25 and R represents a C- alkyl group, a Css cycloalkyl 2 group, a Co aryl group or a 5- or 6-member heteroaryl R1 N R2 group which may have 1 or 2 substituents selected from Substituent group C.: Substituent Group C. wherein R' represents a hydrogen atom, a halogen atom, an 30 Substituent group C. represents the group consisting of a halo amino group, R'—NH- (wherein R' represents a C, gen atom, a C-6 alkyl group, a C1-6 alkoxy group, a Cas alkyl group, a hydroxy Co alkyl group, a C- alkoxy C. cycloalkyl group, a C2-alkenyl group and a C2-alkynyl alkyl group, or a C- alkoxycarbonyl C. alkyl group), group; R’’ (CO) NH (wherein R' represents a C alkyl 35 3 the compound according to item 1 or 2, or the salt group or a C- alkoxy C1-alkyl group), a C- alkyl group, a thereof, wherein a partial structure represented by formula hydroxy Co alkyl group, a cyano C- alkyl group, a C (II): alkoxy group or a C- alkoxy Coalkyl group: R represents a hydrogen atom, a C- alkyl group, an amino group or a diCo alkylamino group: 40 (II) one of X and Y is a nitrogen atom while the other is a (Sy nitrogen atom or an oxygen atom; WN-1 ring A represents a 5- or 6-member heteroaryl ring or a benzene ring which may have 1 or 2 halogenatoms, or 1 or 2 45 C. alkyl groups: in the compound represented by the formula (I) or the formula Z represents a single bond, a methylene group, an ethylene group, an oxygen atom, a Sulfur atom, —CH2O—, OCH , NH CH-NH , NHCH , 50 —CHS , or—SCH -: X (I) R represents ahydrogenatom, ahalogenatom, a Calkyl N group, a C-8 cycloalkyl group, a Co-o aryl group, a 5- or R N (O-ON 71 R3 6-member heteroaryl group, or 5- or 6-member non-aromatic 55 2 heterocyclic group which may have 1 or 2 substituents RI N R2 selected from Substituent group C.: and Substituent Group C. Substituent group C. represents the group consisting of a halo gen atom, a cyano group, a C- alkyl group, a C- alkoxy 60 (I) group, a C- alkoxycarbonyl group, a Css cycloalkyl group. & a C- alkenyl group and a C- alkynyl group (O-ON R3 R" represents a hydrogen atom or a halogen atom; r 71 2 excluding compounds where all of R', R, and R represent 65 R1 N R2 the hydrogen atom at the same time when Z represents the single bond or R represents the hydrogen atom; US 8,158,657 B2 5 6 is a partial structure selected from the group consisting of is a partial structure represented by the following formula (III):

(III) (III)

10 (IV) or a partial structure represented by the following formula (IV): 15

(IV)

(V)

25 6 the compound according to item 1 or 2, or the salt (VI) thereof, wherein X and Y are both nitrogen atoms: 7 the compound according to item 6 or the salt thereof, wherein a partial structure represented by the formula (II): 30 (II)

4 the compound according to item 1 or 2, or the salt 35 thereof, wherein one of X and Y is a nitrogen atom and the other is an oxygen atom; in the compound represented by the formula (I) or the formula 5 the compound according to item 4 or the salt thereof, wherein a partial structure represented by the formula (II): 40 (I) (II) N (Sy Y 3 45 N O-O-s-s WN-1A 4. R2 (I) X in the compound represented by the formula (I) or the formula (I): 50 N SO3-O- - 3

(I) 4. R2 N 55 is a partial structure represented by the following formula S-N-C-O--r 71 (V): 2 R1 N R2 (I) (V) N 60 N SO-Os-R71 2 RI N R2 65 US 8,158,657 B2 7 8 or a partial structure represented by the following formula include all possible isomers, such as structurally possible (VI): geometric isomers, optical isomers generated due to the pres ence of asymmetric carbons, Stereoisomers, tautomers, and mixtures of isomers, and are not limited to formulae being (VI) 5 used for the convenience of description, and may be either one of two isomers or a mixture of both isomers. Thus, the N compounds according to the present invention may be either A. N optically active compounds having an asymmetric carbon atom in their molecules or their racemates, and are not 10 restricted to either of them but include both. Furthermore, the compounds according to the present invention may exhibit crystalline polymorphism, but likewise are not restricted to 8 the compound according to any one of items 1 to 7 or any one of these, but may be in any one of these crystal forms the Salt thereof, wherein R represents an amino group; 15 or exist as a mixture of two or more crystal forms. The 9 the compound according to item 8 or the salt thereof, compounds according to the present invention also include wherein R' represents a hydrogen atom, an amino group or a both anhydrous and Solvates such as hydrated forms. Calkoxy C. alkyl group: The term "C. alkyl group' used in the present specifica 10 the compound according to any one of items 1 to 7 or tion refers to a straight-chain or branched-chain alkyl group the salt thereof, wherein R' represents an amino group andR 20 with 1 to 6 carbon atoms which is a monovalent group represents a hydrogen atom; induced by removal of any one hydrogen atom from an ali 11 the compound according to any one of items 1 to 10 phatic hydrocarbon with 1 to 6 carbon atoms. Specifically, or the salt thereof, wherein the ring A represents a pyridine examples of "Ce alkyl group' includes a methyl group, an ring, a benzene ring, afuran ring, a thiophene ring or a pyrrole ethyl group, a n-propyl group, an isopropyl group, a n-butyl ring: 25 group, an isobutyl group, a sec-butyl group, a tert-butyl 12 the compound according to item 11 or a salt thereof, group, a n-pentyl group, an isopentyl group, a sec-pentyl wherein ring A represents a pyridine ring or a benzene ring; group, a neopentyl group, a 1-methylbutyl group, a 2-meth 13 the compound according to any one of items 1 to 12 ylbutyl group, a 1,1-dimethylpropyl group, a 1,2-dimethyl or the salt thereof, wherein Z represents an oxygen atom, propyl group, a n-hexyl group, an isohexyl group, a 1-meth —CHO— or —OCH : 30 ylpentyl group, a 2-methylpentyl group, a 3-methylpentyl 14 a pharmaceutical composition comprising the com group, a 1,1-dimethylbutyl group, a 1,2-dimethylbutyl group, pound according to any one of items 1 to 13 or the salt a 2.2-dimethylbutyl group, a 1,3-dimethylbutyl group, a 2.3- thereof. dimethylbutyl group, a 3.3-dimethylbutyl group, a 1-ethylbu 15 a medicament comprising the compound according to tyl group, a 2-ethylbutyl group, a 1.1.2.-trimethylpropyl any one of items 1 to 13 or the salt thereof; 35 group, a 1.2.2-trimethylpropyl group, a 1-ethyl-1-methylpro 16 an antifungal agent comprising the compound according pyl group, a 1-ethyl-2-methylpropyl group or the like, pref to any one of items 1 to 13 or the salt thereof, as an active ingredient; erably a methyl group, an ethyl group, a n-propyl group, an 17 a method for preventing and/or treating a fungal infec isopropyl group, a n-butyl group, an isobutyl group, a sec tion comprising administering a pharmacologically effective 0 butyl group or a tert-butyl group or the like. dose of the compound according to any one of items 1 to The term "Ce alkenyl group' used in the present specifi 13 or the salt thereof; cation refers to a straight-chain or branched-chain alkenyl 18 a use of the compound according to any one of items 1 group with 2 to 6 carbon atoms which may contain 1 or 2 to 13 or the salt thereof for manufacturing an antifungal double bonds. Specifically, examples of "Coalkenyl group' agent. 45 include an ethenyl group, a 1-propenyl group, a 2-propenyl group, a 1-butenyl group, a 2-butenyl group, a 3-butenyl ADVANTAGEOUSEFFECTS OF THE group, a 2-methyl-1-propenyl group, a pentenyl group, a INVENTION 3-methyl-2-butenyl group, a hexenyl group, a hexanedienyl group or the like, preferably an ethenyl group, a 1-propenyl The compound (I) of the present invention or a salt thereof 50 group, a 2-propenyl group, a 1-butenyl group, a 2-butenyl 1) acts against the onset, development and persistence of group, a 3-butenyl group, a 2-methyl-1-propenyl group, a infections by inhibiting fungal GPI biosynthesis, thereby 3-methyl-2-butenyl group or the like. inhibiting expression of cell wall proteins and blocking cell The term "Ce alkynyl group' used in the present specifi wall assembly while preventing the fungus from attaching to cation refers to a straight-chain or branched-chain alkynyl cells So that the pathogen cannot become pathogenic, and 2) 55 chain with 2 to 6 carbonatoms which may contain 1 or 2 triple is Superior in terms of physical properties, safety and meta bonds. Specifically, examples of “Calkynyl group” include bolic stability, and is extremely useful as a preventive or an ethynyl group, a 1-propynyl group, a 2-propynyl group, a therapeutic agent for fungal infections. 1-butynyl group, a 2-butynyl group, a 3-butynyl group, a pentynyl group, a hexynyl group, a hexanediynyl group or the DETAILED DESCRIPTION OF THE INVENTION 60 like, preferably an ethynyl group, a 1-propynyl group, a 2-propynyl group, a 1-butynyl group, a 2-butynyl group, a The present invention is explained below in more detail by 3-butynyl group or the like. reference to the symbols and the terms used herein being The term “Cs cycloalkyl group' used in the present speci defined and the following examples. fication refers to a cyclic aliphatic hydrocarbon group with 3 Herein, a structural formula of a compound sometimes 65 to 8 carbonatoms. Specifically, examples of "Cs cycloalkyl represents a certain isomer for convenience of description. group' include a cyclopropyl group, a cyclobutyl group, a However, compounds according to the present invention may cyclopentyl group, a cyclohexyl group, a cycloheptyl group, US 8,158,657 B2 10 a cyclooctyl group or the like, preferably a cyclopropyl group, The term “halogen atom' used in the present specification a cyclobutyl group, a cyclopentyl group, a cyclohexyl group refers a fluorine atom, a chlorine atom, a bromine atom or an or the like. iodine atom. The term "Ce alkoxy group' used in the present specifi The term "hetero atom' used in the present specification cation refers to a group in which an oxygenatom is bonded to refers to a nitrogen atom, a Sulfur atom or an oxygen atom. terminus of the “C. alkyl group' defined above. Specifi The term “5- or 6-member heteroaryl ring used in the cally, examples of “C. alkoxy group' include a methoxy present specification refers to an aromatic ring in which the group, an ethoxy group, a n-propoxy group, an isopropoxy number of atoms making up the ring is 5 or 6, and 1 or more group, a n-butoxy group, an isobutoxy group, a sec-butoxy hetero atoms are included in the atoms making up the ring. group, a tert-butoxy group, a n-pentyloxy group, an isopen 10 tyloxy group, a sec-pentyloxy group, a neopentyloxy group, a Specifically, examples of “5- or 6-member heteroaryl ring 1-methylbutoxy group, a 2-methylbutoxy group, a 1,1-dim include a furan ring, a thiophene ring, a pyrrole ring, a pyri ethylpropoxy group, a 1,2-dimethylpropoxy group, a n-hexy dine ring, a pyrazine ring, a pyridazine ring, a pyrimidine loxy group, an isohexyloxy group, a 1-methylpentyloxy ring, a triazole ring (a 1,2,3-triazolering, a 1,2,4-triazole ring, group, a 2-methylpentyloxy group, a 3-methylpentyloxy 15 etc.), a tetrazole ring (a 1H-tetrazole ring, a 2H-tetrazole ring, group, a 1,1-dimethylbutoxy group, a 1,2-dimethylbutoxy etc.), a thiazole ring, a pyrazole ring, an oxazole ring, an group, a 2,2-dimethylbutoxy group, a 1,3-dimethylbutoxy isoxazole ring, an isothiazole ring, an oxadiazole ring, a thia group, a 2,3-dimethylbutoxy group, a 3.3-dimethylbutoxy diazole ring or the like. group, a 1-ethylbutoxy group, a 2-ethylbutoxy group, a 1.1, The term “5- or 6-member heteroaryl group' used in the 2-trimethylpropoxy group, a 1.2.2-trimethylpropoxy group, a present specification refers to a monovalent group induced by 1-ethyl-1-methylpropoxy group, a 1-ethyl-2-methylpropoxy removing 1 hydrogen atom from any position in an aromatic group or the like, preferably a methoxy group, an ethoxy ring in which the number of atoms making up the ring is 5 or group, a n-propoxy group, an isopropoxy group, a n-butoxy 6 and 1 or more hetero atoms are included in the atoms group, an isobutoxy group, a sec-butoxy group, a tert-butoxy making up the ring. Specifically, examples of '5- or 6-mem group or the like. 25 ber heteroaryl group' include a furyl group (a 2-furyl group or The term “hydroxyl C alkyl group' used in the present a 3-furyl group, etc.), a thienyl group (a 2-thienyl group or a specification refers to a group in which any of the hydrogen 3-thienyl group, etc.), a pyrrolyl group (a 1-pyrrolyl group, a atoms in a "Ce alkyl group’ as defined above has been 2-pyrrolyl group or a 3-pyrrolyl group, etc.), a pyridyl group replaced by a hydroxyl group. Specifically, examples of (a 2-pyridyl group, a 3-pyridyl group, a 4-pyridyl group, etc.), “hydroxyl C alkyl group” include a hydroxymethyl group, 30 a pyrazinyl group, a pyridazinyl group (a 3-pyridazinyl group a 1-hydroxyethyl group, a 2-hydroxyethyl group, a 1-hy or a 4-pyridazinyl group, etc.), a pyrimidinyl group (a 2-py droxy-n-propyl group, a 2-hydroxy-n-propyl group, a 3-hy rimidinyl group, a 4-pyrimidinyl group or a 5-pyrimidinyl droxy-n-propyl group, a 1-hydroxy-isopropyl group, a 2-hy group, etc.), a triazolyl group (a 1,2,3-triazolyl group or a droxy-isopropyl group, a 3-hydroxy-isopropyl group, a 1,2,4-triazolyl group, etc.), a tetrazolyl group (a 1H-tetrazolyl 1-hydroxy-tert-butyl group or the like, preferably a 35 group or a 2H-tetrazolyl group, etc.), a thiazolyl group (a hydroxymethyl group, a 1-hydroxyethyl group, a 2-hydroxy 2-thiazolyl group, a 4-thiazolyl group or a 5-thiazolyl group, ethyl group or the like. etc.), a pyrazolyl group (a 3-pyrazolyl group or a 4-pyrazolyl The term "Ce alkoxycarbonyl group' used in the present group, etc.), an oxazolyl group (a 2-oxazolyl group, a 4-OX specification refers to a group in which a carbonyl group is azolyl group or a 5-oxazolyl group, etc.), an isoxazolyl group bonded to terminus of the "Calkoxy group' defined above. 40 (a 3-isoxazolyl group, a 4-isoxazolyl group or a 5-isoxazolyl Specifically, examples of "Ce alkoxycarbonyl group' group, etc.), an isothiazolyl group (a 3-isothiazolyl group, a include a methoxycarbonyl group, an ethoxycarbonyl group, 4-isothiazolyl group or a 5-isothiazolyl group, etc.), an oxa a n-propoxycarbonyl group, an isopropoxycarbonyl group or diazolyl group, a thiadiazolyl group or the like. the like. The term “5- or 6-member non-aromatic heterocyclic The term "Ce alkoxycarbonyl C. alkyl group' used in 45 group' used in the present specification refers to a monova the present specification refers to a group in which the “C. lent group induced by removing 1 hydrogen atom from any alkyl group' defined above is bonded to terminus of the “C. position in a non-aromatic ring in which the number of atoms alkoxycarbonyl group' defined above. Specifically, examples making up the ring is 5 or 6 and 1 or more hetero atoms are of the "Calkoxycarbonyl Calkyl group” include a meth included in the atoms making up the ring. Specifically, oxycarbonyl methyl group, a methoxycarbonyl ethyl group, 50 examples of “5- or 6-member non-aromatic heterocyclic an ethoxycarbonyl methyl group, an ethoxycarbonyl ethyl group' include a pyrrolidinyl group, a piperadinyl group, a group or the like. piperidinyl group, a morpholinyl group, a tetrahydrofuryl The term "Coo aryl group' used in the present specifica group, a tetrahydropyranyl group or the like. tion refers to an aromatic hydrocarbon cyclic group with 6 to The term "di C-alkylamino group' used in the present 10 carbonatoms. Specifically, examples of "Coo aryl group' 55 specification refers to a group in which 2 hydrogen atoms of include a phenyl group, a 1-naphthyl group, a 2-naphthyl the amino group are replayed with the “C. alkyl groups” group, an indenyl group, an azulenyl group, a heptalenyl defined above being the same as or different from each other. group or the like, preferably a phenyl group, a 1-naphthyl Specifically, examples of the term “di C-alkylamino group' group, 2-naphthyl group or the like. include a N,N-dimethylamino group, a N,N-diethylamino The term "Calkoxy Calkyl group' used in the present 60 group, a N,N-di-n-propylamino group, a N,N-di-isopropy specification refers to a group in which any of the hydrogen lamino group, a N,N-di-n-butylamino group, a N,N-isobuty atoms in a "Ce alkyl group’ as defined above has been lamino group, a N,N-di-sec-butylamino group, a N,N-di-tert replaced by a "Calkoxy group’ as defined above. Specifi butylamino group, a N-ethyl-N-methylamino group, a N-n- cally, examples of "Ce alkoxy C. alkyl group' include a propylamino-N-methylamino group, a N-isopropyl-N- methoxymethyl group, an ethoxymethyl group, a n-pro 65 methylamino group, a N-n-butyl-N-methylamino group, a poxymethyl group, a methoxyethyl group, an ethoxyethyl N-isobutyl-N-methylamino group, a N-sec-butyl-N-methy group or the like. lamino group, a N-tert-butyl-N-methylamino group or the US 8,158,657 B2 11 like, preferably a N,N-dimethylamino group, a N,N-diethy lamino group, N-ethyl-N-methylamino group or the like. The term “may have 1 or 2 substituents’ used in the speci fication means that there may be 1 or 2 substituents in any combination in sites capable of Substituting. R" preferably represents a hydrogenatom, a halogenatom, an amino group, a C- alkyl group, a C- alkoxy group, a C. alkylamino group, a hydroxyl C alkylamino group, or a C- alkoxy C. alkyl group, and more preferably a hydro It is preferable that one of X and Y be a nitrogen atom and gen atom, an amino group or a C- alkoxy Co alkyl group. 10 the other be an oxygenatom, or that both X and Y be nitrogen with a methoxymethyl group being preferred as the C atoms, and when one of X and Y is a nitrogen atom and the alkoxy C. alkyl group. other is an oxygenatom, the partial structure which contains R represents a hydrogen atom, an amino group or di C. X and Y, and which is represented by the following formula alkylamino group, with a hydrogen atom or an amino group (II): being preferred. 15 One of X and Y is a nitrogen atom while the other is a nitrogen atom or an oxygen atom. (II) The partial structure which contains X and Y and which is X represented by formula (II) below: (SyN WN-1 (II)

25 has a structure such as that shown by formulae (III) or (IV) below, preferably with the left end bound to the 3-position of a pyridine ring via a single bond and the right end linked to an has a structure such as those shown below, preferably with the left side bound to the 3-position of a pyridine ring via a single 30 A ring via a methylene group: bond, and the right side bound to an A ring via a methylene group: (III)

(III) 35 ON

N \ | /ox O 40

(IV) (IV)

N 45 /cy (V) 50 while if X and Y are both nitrogenatoms, the partial structure which contains X and Y, and which is represented by the following formula (II):

55 O (II) (VI) X N (Syf WN-1/ 60

has a structure such as that shown by formula (V) or (VI) In the case of the partial structure of formula (III), for 65 below, preferably with the left end bound to the 3-position of example, the structure of the compound of the present inven a pyridine ring via a single bond and the right end bound to an tion is shown by the following formula: A ring via a methylene group:

US 8,158,657 B2 15 16 3-(3-(6-phenoxymethyl-pyridin-3-ylmethyl)-isoxazol-5-yl)- able salt thereof with a diluent, and if required, a binder, a pyridin-2,6-diamine; disintegrating agent, a lubricant, a colorant, a flavorant or the 3-(3-(4-(5-fluoro-pyridin-2-ylmethoxy)-benzyl)-isoxazol-5- like, and formulating the mixture into powders, fine granules, yl)-pyridin-2,6-diamine; granules, tablets, coated tablets, capsules or the like accord 3-(3-(4-(6-fluoro-pyridin-2-ylmethoxy)-benzyl)-isoxazol-5- ing to the conventional methods. yl)-pyridin-2,6-diamine; Examples of the materials include animal and vegetable 3-(3-(1-benzyl-1H-pyrrol-3-ylmethyl)-isoxazol-5-yl)-pyri oils such as Soybean oil, beef tallow, and synthetic glyceride; din-2,6-diamine; hydrocarbons such as liquid paraffin, squalane, and solid 3-(3-(6-(4-fluoro-benzyloxy)-pyridin-3-ylmethyl)-isoxazol paraffin; ester oils such as octyldodecyl myristate and iso 5-yl)-pyridin-2,6-diamine; 10 propyl myristate; higher alcohols such as cetostearyl alcohol 3-(3-(4-(5-chloro-furan-2-ylmethyl)-benzyl)-isoxazol-5-yl)- and behenyl alcohol; silicone resins; silicone oils; Surfactants pyridin-2,6-diamine; Such as polyoxyethylene fatty acids ester, Sorbitan fatty acid 3-(3-(6-(3-fluoro-phenoxy)pyridin-3-ylmethyl)-isoxazol-5- ester, polyoxyethylene Sorbitan fatty acid ester, polyoxyeth yl)-pyridin-2,6-diamine; ylene hydrogenated castor oil, and polyoxyethylene polyox 3-(3-(4-phenylaminomethyl-benzyl)-isoxazol-5-yl)-pyridin 15 ypropylene block co-polymer, water-soluble polymers such 2,6-diamine; as hydroxyethyl cellulose, polyacrylic acid, carboxyvinyl 3-(3-(6-(4-fluoro-phenoxy)-pyridin-3-ylmethyl)-isoxazol-5- polymer, polyethylene glycol, polyvinylpyrrolidone, and yl)-pyridin-2,6-diamine; methyl cellulose; lower alcohols such as ethanol and isopro 3-(3-(4-(thiazol-2-ylmethoxy)-benzyl)-isoxazol-5-yl)-pyri panol; polyhydric alcohols such as glycerol, propylene gly din-2,6-diamine; col, dipropylene glycol, and Sorbitol; Sugars Such as glucose 3-(3-(5-(4-fluoro-phenoxy-thiophen-2-ylmethyl)-isoxazol and Sucrose; inorganic powder Such as anhydrous silicic acid, 5-yl)-pyridin-2,6-diamine; magnesium aluminum silicate, and aluminum silicate; and 6-methoxymethyl-3-(3-(4-(pyridin-2-ylmethoxy)-benzyl)- pure water. Examples of the diluents include lactose, corn isoxazol-5-yl)-pyridin-2-ylamine; starch, white Sugar, glucose, mannitol, Sorbitol, crystalline 6-methyl-3-(3-(4-pyridin-2-yloxymethyl)-benzyl)-isoxazol 25 cellulose, silicon dioxide or the like. Examples of the binders 5-yl)-pyridin-2-ylamine; include polyvinyl alcohol, polyvinyl ether, methylcellulose, 5-(3-(4-pyridin-2-yloxymethyl)-benzyl)-isoxazol-5-yl)-py ethylcellulose, gum Arabic, tragacanth, gelatin, shellac, ridin-2-ylamine; hydroxypropyl methylcellulose, hydroxypropyl cellulose, 3-(1-(4-(pyridin-2-ylmethoxy)-benzyl)-1H-pyrazol-4-yl)- polyvinylpyrrolidone, polypropylene glycol-polyoxyethyl pyridin-2-ylamine; and 30 ene block co-polymer, and meglumine or the like. Examples 3-(3-(4-(pyridin-2-yloxymethyl)-benzyl)-isoxazol-5-yl)-py of disintegrating agents include starch, agar, gelatin powder, ridine. crystalline cellulose, calcium carbonate, sodium hydrogen Examples of the term “salt used in the present specifica carbonate, calcium citrate, dextrin, pectin, calcium car tion include a salt with an inorganic acid, a salt with an boxymethyl cellulose or the like. Examples of lubricants organic acid, a salt with an inorganic base, a salt with an 35 include magnesium Stearate, talc, polyethylene glycol, silica, organic base, a salt with an acidic or basic amino acid or the hydrogenated vegetable oil or the like. Examples of colorants like. Among these salts, it is preferable that a salt used herein include those pharmaceutically acceptable. Examples of fla be a pharmaceutically acceptable. Vorants include cocoa powder, peppermint camphor, aro Preferable examples of the salt with the inorganic acid matic powder peppermint oil, Borneo camphor, cinnamon include salts with hydrochloric acid, hydrobromic acid, sul 40 powder or the like. Tablets and granules may be coated with furic acid, nitric acid, phosphoric acid or the like. Preferable Sugar, or if required, other appropriate coatings can be made. examples of the salt with the organic acid include salts with Solutions, such as syrups or injectable preparations, to be acetic acid, Succinic acid, fumaric acid, maleic acid, tartaric administered can be formulated by combining a compound acid, citric acid, lactic acid, Stearic acid, benzoic acid, meth according to the present invention or a pharmaceutically anesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid 45 acceptable salt thereof with a pH modulator, a solubilizing or the like. agent, an isotonizing agent or the like, and ifrequired, with an Preferable examples of the salt with the acidic amino acid auxiliary Solubilizing agent, a stabilizer or the like, according include salts with aspartic acid, glutamic acid or the like. to the conventional methods. Methods for manufacturing Preferable examples of the salt with the basic amino acid external preparations are not limited and Such preparations include salts with arginine, lysine, ornithine or the like. 50 can be manufactured by the conventional methods. Specifi The term “antifungal agent used in the present specifica cally, various materials typically used for manufacturing tion refers to a preventive agent or a therapeutic agent for pharmaceuticals, quasi drugs, cosmetics or the like can be fungal infection. used as base materials for the external formulation. More The compounds according to the present invention, or salts specifically, examples of base materials to be used include or hydrates thereof, can be formulated into tablets, powders, 55 animal and vegetable oils, minerals oils, ester oils, wax, fine granules, granules, coated tablets, capsulates, syrups, higher alcohols, fatty acids, silicone oil, Surfactants, phos troches, inhalants, Suppositories, injections, ointments, eye pholipids, alcohols, polyhydric alcohols, water-soluble poly ointments, tapes, eye drops, nose drops, ear drops, cata mers, clay minerals, pure water or the like. Furthermore, plasms, lotions or the like, by the conventional methods. external preparations of the present invention can contain, as Such formulation can be achieved by using typical dilu 60 required, pH modulators, antioxidants, chelating agents, anti ents, binders, lubricants, colorants, flavorants, and, as neces bacterial/antifungal agents, colorants, odoriferous Substances sary, stabilizers, emulsifiers, absorbefacients, Surfactants, pH or the like. But this does not limit the type of base materials modulators, preservatives, antioxidants or the like, and mate that are to be used in the external preparations of the present rials commonly used as ingredients of pharmaceutical prepa invention. If required, the preparation may contain differen rations according to the conventional methods. For example, 65 tiation inducers, blood flow improving agents, antimicrobial an oral preparation can be produced by combining a com agents, antiphologistics, cell activators, vitamins, amino pound of the present invention or a pharmaceutically accept acids, humectants, keratolytic agents or the like. The amount US 8,158,657 B2 17 18 of the base materials listed above is adjusted within a concen Manufacturing Method 1-1 Method for Manufacturing tration range used for producing typical external prepara Compound (1a): tions. When administering the compound of the present inven tion or a salt thereof, the forms of the compounds are not 2 limited in particular, and the compound can be given orally or N parenterally by the conventional method. For instance, the -- compound can be administered as a dosage form such as R1 % R2 tablets, powders, granules, capsules, syrups, troches, inhal ants, suppositories, injections, ointments, eye ointments, 10 (1b) tapes, eye drops, nasal drops, ear drops, cataplasms and C ZN lotions. Dose of a medicament according to the present invention can be selected appropriately according to symptom severity, 15 HO1 age, sex, body weight, forms of administration, type of salts, (1c) specific type of disease or the like. The does varies remarkably depending on the patients disease, symptom severity, age and sex, drug Susceptibility or the like. An oral preparation according to the present inven tion can be generally administered once or several time at a dose of from 1 to 10000 mg/adult/day, preferably from 10 to 2000 mg/adult/day. An injection according to the present invention can be generally administered at a dose of from 0.1 to 10000 mg/adult/day, preferably from 1 to 2000 mg/adult/ 25 day. (1a) General Synthesis Methods The method for manufacturing the compounds represented (wherein the ring A. R. R. R. and Z have the same mean by formula (I) according to the present invention (hereinafter 30 ings as defined above.) referred to as compounds (I)) is discussed here. The com pounds according to the present invention can be synthesized Compound (1b) which is a commercially available product by ordinary organic synthesis methods, but for example, can be used as is or compound (1b) can also be manufactured among the compounds (I), the compounds represented by from a commercially available product by the well known formula (1a), formula (2a), formula (3a), formula (4a), for 35 methods. In addition, compound (1b) can be manufactured by mula (5a), formula (6a-1), formula (6a-3), formula (7), for the methods described in the Manufacturing Examples in the mula (8a), formula (9a) and formula (10a) (hereinafter Examples or according to Manufacturing Method 1-2-1 or referred to as compound (1a), compound (2a), compound the like. (3.a), compound (4a), compound (5a), compound (6a-1), Compound (1c) can be manufactured by the well known compound (6a-3), compound (7a), compound (8a), com 40 methods from a commercial available product. Compound pound (9a) and compound (10a), respectively) can be synthe (1c) can also be manufactured by the methods described in the sized by the methods given Manufacturing Method 1 to Manufacturing Examples in the Examples or according to Manufacturing Method 10 below and the like. Manufacturing Method 1-3-1 and the like. Manufacturing Method 1 Typical Method for Manufactur ing Compound (1a): 45 Step 1 This step is a step wherein compound (1a) is obtained by reacting compound (1b) and compound (1c) in the presence of a base. There are no particular limitations on the solvent used in this reaction as long as it dissolves the starting mate 50 rials to a certain extent without impeding the reaction. (I) Examples of the solvents used in this reaction include ether Solvents such as tetrahydrofuran and diethyl ether, aromatic hydrocarbon solvents such as benzene and toluene; amide solvents such as N,N-dimethylformamide and N-methylpyr 55 rolidinone; alcohol Solvents such as methanol and ethanol: and water, methylene chloride, chloroform, ethyl acetate, dimethyl sulfoxide, mixed solvents of the foregoing and the like. Examples of the base used in this reaction include tri 60 ethylamine, N,N-diisopropylethylamine, Sodium hydrogen carbonate, potassium carbonate and the like. Compound (1c) can be used in the amount of 1 to 3 equivalents, preferably 1 to 2 equivalents, based on compound (1b). The base can be used in the amount of 1 to 3 equivalents based on compound 65 (1c). The reaction temperature is from room temperature to wherein ring9. A. R. R. R. and R and Z are defined as reflux temperature, and the reaction time is from 10 minutes above.) to 24 hours. US 8,158,657 B2 19 20 Manufacturing Method 1-2-1 Method 1 for Manufacturing The phosphine ligand is used in the amount of 0.01 to 1.2 Compound (1b): equivalents based on compound (1b-1). The copper catalyst is used in the amount of 0.001 to 0.3 equivalents based on compound (1b-1). The reaction temperature is from room N Hal temperature to reflux temperature, and the reaction time is from 30 minutes to 24 hours. 2 step 1-1 Step 1-2 RI N R2 This step is a step wherein compound (1b) is obtained by reacting compound (1b-2) with a base. There are no particular (1b-1) 10 R5 limitations on the solvent used in this reaction as long as it dissolves the starting materials to a certain extent without Si-R impeding the reaction. Examples of the solvents in this step 2^ Y. include ether solvents such as tetrahydrofuran and diethyl N step 1-2 ether; alcohol solvents such as methanol and ethanol; amide 15 solvents such as N,N-dimethylformamide and N-methylpyr 2 R1 N R2 rolidinone; and acetonitrile, dimethylsulfoxide, water, mixed solvents of the foregoing and the like. Examples of the base include potassium carbonate, sodium hydroxide, tetrabuty lammonium fluoride, potassium fluoride, cesium fluoride and the like. The base is used in the amount of 0.05 to 10 equiva lents based on compound (1b-2). The reaction temperature is from 0°C. to reflux temperature, and the reaction time is from 5 minutes to 24 hours. (1b) 25 Manufacturing Method 1-2-2 Method 2 for Manufacturing Compound (1b): (wherein R' and R are defined as above, Hal represents a halogen atom, and R and R represent each independently C. alkyl groups.) O Compound (1b-1) which is a commercially available prod 30 uct can be used as is, or compound (1b-1) can also be manu N OH Hestep1-3 factured from commercially available products by the well 2 known methods. R1 N R2 Step 1-1 35 (1b-3) This step is a step wherein compound (1b-2) is obtained by O reacting compound (1b-1) with an ethynyl silane derivative. R7 Compound (1b-2) can be obtained by reacting compound N o1 Her (1b-1) with an ethynyl silane derivative in the presence of a step 1-4 palladium catalyst, a base and a copper catalyst. A phosphine 40 2 ligand may also be added to obtain good results. There are no R1 N R2 particular limitations on the solvent used in this reaction as (1b-4) long as it can dissolve the starting materials to a certain extent N OH without impeding the reaction. Examples of the solvents used in this reaction include ether solvents such as tetrahydrofuran 45 step1-5 and 1,4-dioxane; amide solvents such as N,N-dimethylfor RI 4. R2 mamide and N-methylpyrrolidinone; and acetonitrile, dim (1b-5) ethyl sulfoxide, mixed solvents of the foregoing and the like. O Examples of the ethynyl silane derivative include trimethyl silylacetylene, triethylsilylacetylene, triisopropylsilylacety 50 2 lene, t-butyldimethylsilylacetylene and the like. Examples of r --step 1-6 the palladium catalysts include palladium(II) acetate, tetrakis 2 2 (triphenylphosphine)palladium (O), dichlorobis(triph RI N R2 RI N R2 enylphosphine)palladium (II), dichlorobis(tri-o-tolylphos (1b-6) (1b) phine)palladium (II), bis(tri-t-butylphosphine)palladium (O), 55 or tris(dibenzylideneacetone)dipalladium (O) and the like. Examples of the base include triethylamine, N,N-diisopropy (wherein R' and Rare defined as above, and R7 represents a lethylamine, pyridine and the like. Examples of the phosphine C- alkyl group.) ligand include triphenylphosphine, tri-o-tolylphosphine, tri Compound (1b-3) which is a commercially available prod t-butylphosphine and the like. A copper catalyst can be added 60 uct can be used as is, or compound (1b-3) can also be manu in this reaction. Examples of the copper catalyst include factured from commercially available products by the well copper, copper (I) iodide, copper (I) bromide, copper (I) known methods. chloride and the like. The ethynyl silane derivative is used in Step 1-3 the amount of 1 to 5 equivalents based on compound (1b-1). This step is a step wherein compound (1b-4) is obtained by The palladium catalyst is used in the amount of 0.01 to 0.3 65 esterifying compound (1b-3) in the presence of an acid. The equivalents based on compound (1b-1). The base is used in solvent used in this reaction is preferably an alcohol solvent the amount of 2 to 5 equivalents based on compound (1b-1). Such as methanol, ethanol and the like. Examples of the acids US 8,158,657 B2 21 22 include sulfuric acid, hydrochloric acid, hydrobromic acid temperature, with a reaction time being from 10 minutes to 48 and the like. The acid can be used in the amount from a hours. The reaction temperature for reacting the alcohol is catalytic amount to a solvent amount based on compound from 0°C. to reflux temperature, with a reaction time being (1b-3). The reaction temperature is from room temperature to from 10 minutes to 48 hours. The alcohol can be used as the reflux temperature, and the reaction time is from 1 hour to 72 Solvent in this reaction. In this case, compound (1b-4) can be hours. obtained by adding the halogenating agent to a mixture of the Compound (1b-4) can also be obtained from compound solvent and compound (1b-3). The reaction temperature is (1b-4) by the methods described as Alternative Methods (1), from 0°C. to room temperature, and the reaction time is from (2) and (3) below. 10 minutes to 24 hours. Alternative Method (1): Compound (1b-4) can be con 10 Step 1-4 Verted into a methyl ester derivative using diazomethane or This step is a step wherein compound (1b-5) is obtained by trimethylsilyl diazomethane. There are no particular limita reduction of compound (1b-4). There are no particular limi tions on the solvent used in this reaction as long as it dissolves tations on the solvent used in this reaction as long as it dis the starting materials to a certain extent without impeding the Solves the starting materials to a certain extent without imped reaction. Examples of the solvent used in this reaction include 15 ing the reaction, buttetrahydrofuran is preferred. Examples of ether solvents such as tetrahydrofuran and diethyl ether; aro the reducing agent in this reaction include lithium aluminum matic hydrocarbon Solvents such as benzene and toluene; hydride, lithium aluminum hydride-aluminum chloride (alu alcohol solvents such as methanol and ethanol; and methyl minum chloride in the amount of 1 to 1.5 equivalents based on ene chloride, hexane, mixed solvents of the foregoing and the lithium aluminum hydride), lithium borohydride and the like. like. The diazomethane or trimethylsilyl diazomethane is The reducing agent is used in the amount of 0.5 to 4 equiva used in the amount of 1 to 2 equivalents based on compound lents based on compound (1b-4). The reaction temperature is (1b-3). The reaction temperature is from 0°C. to room tem from 0°C. to reflux temperature, and the reaction time is from perature, and the reaction time is from 10 minutes to 24 hours. 10 minutes to 48 hours. Alternative Method (2): Compound (1b-3) can be con Step 1-5 Verted into compound (1b-4) using an alkylating agent in the 25 This step is a step wherein compound (1b-6) is obtained by presence of a base. There are no particular limitations on the oxidation of compound (1b-5). There are no particular limi Solvent used in this reaction as long as it dissolves the starting tations on the solvent used in this reaction as long as it dis materials to a certain extent without impeding the reaction. Solves the starting materials to a certain extent without imped Examples of the solvents used in this reaction include ether ing the reaction. Examples of the solvents used in this Solvents such as tetrahydrofuran and diethyl ether, aromatic 30 reaction include ether solvents such as tetrahydrofuran and hydrocarbon solvents such as benzene and toluene; amide diethyl ether; aromatic hydrocarbon solvents such as benzene solvents such as N,N-dimethylformamide and N-methylpyr and toluene: alcohol solvents such as methanol and ethanol: rolidinone; alcohol Solvents such as methanol and ethanol: and methylene chloride, acetone, hexane, mixed solvents of and water, acetone, acetonitrile, dimethyl Sulfoxide, mixed the foregoing and the like. Examples of oxidizing agent used Solvents of the foregoing and the like. A phase-transfer cata 35 in this reaction include manganese dioxide, pyridinium chlo lyst Such as tetrabutylammonium bromide can also be added rochromate, pyridinium dichromate, dimethyl Sulfoxide-ac to this reaction. Examples of the base used in this reaction tivator, tetrapropylammonium perruthenate, dichlorotris include potassium hydroxide, Sodium hydroxide, lithium (triphenylphosphine)ruthenium (II), 1,1,1-tris(acetyloxy)-1, hydroxide, potassium carbonate, cesium carbonate, cesium 1-dihydro-1,2-benziodoxol-3-(1H)-on (Dess-Martin fluoride and the like. Examples of the alkylating agents 40 Periodinane) and the like. The oxidizing agent is used in the include iodomethane, iodoethane, dimethylsulfate and the amount of from the catalytic amount to 20 equivalents based like. The base is used in the amount of 1 to 1.5 equivalents on compound (1b-5). When oxidizing with dimethyl sulfox based on compound (1b-3). The alkylating agent is used in the ide-activator, examples of the activator include acid anhy amount of 1 to 2 equivalents based on compound (1b-3). The drides such as acetic anhydride and trifluoroacetic anhydride; reaction temperature is from 0°C. to reflux temperature, and 45 acid chlorides such as oxalyl chloride and thionyl chloride: the reaction time is from 1 hour to 72 hours. and chlorine, N-chlorosuccinimide and the like. The dimethyl Alternative Method (3): Compound (1b-3) can be con sulfoxide is used in the amount of 1 to 20 equivalents based on Verted into an acid chloride using a halogenating agent, and the activator. When using tetrapropyl ammonium perruthen then converted into compound (1b-4) by addition of alcohol. ate or dichlorotris(triphenylphosphine)ruthenium (II) in a There are no particular limitations on the solvent used in this 50 catalytic amount, an oxidizing agent such as N-methylmor reaction as long as it dissolves the starting materials to a pholine-N-oxide orbis(trimethylsilyl)peroxide can be used at certain extent without impeding the reaction. Examples of the the same time. The reaction temperature is from -78°C. to Solvents used in this reaction include aromatic hydrocarbon reflux temperature, and the reaction time is from 10 minutes Solvents such as benzene and toluene; amide solvents such as to 72 hours. N,N-dimethylformamide and N-methylpyrrolidinone; and 55 Step 1-6 acetonitrile, methylene chloride, 1,2-dichloroethane, mixed This step is a step wherein compound (1b) is obtained from Solvents of the foregoing and the like. The halogenating agent compound (1b-6) in the presence of a base using a diazo can also be used as the solvent. A catalytic amount of pyridine compound. Examples of the diazo compound used in this or a phase-transfer catalyst Such as benzyltriethylammonium reaction include trimethylsilyl diazomethane, (1-diazo-2- chloride can also be added to this reaction. Examples of the 60 Oxopropyl)-phosphoric acid dimethyl ester, diazomethyl halogenating agents include thionyl chloride, phosphorus phosphoric acid dimethyl ester and the like. There are no pentachloride and the like. Examples of the alcohols include particular limitations on the solvent used in this reaction as methanol, ethanol and the like. The halogenating agent is long as it dissolves the starting materials to a certain extent used in the amount of 1 to 20 equivalents based on compound without impeding the reaction. Examples of the solvents used (1b-3). The alcohol is used in the amount of 1 to 20 equiva 65 in this reaction include ether solvents such as tetrahydrofuran lents based on compound (1b-3). The reaction temperature and diethyl ether, aromatic hydrocarbon Solvents such as during conversion to an acid chloride is from 0°C. to reflux benzene and toluene; alcohol solvents such as methanol and US 8,158,657 B2 23 24 ethanol; and methylene chloride, hexane, mixed solvents of Manufacturing Method 1-2-3 Method for Manufacturing the foregoing and the like. When using trimethylsilyl diaz Compound (1b-3): omethane as the diazo compound, n-butyl lithium and lithium diisopropylamide can be used as the base. When using a phosphoric acid ester derivative Such as (1-diazo-2-oxopro O pyl)-phosphoric acid dimethyl ester and diazomethyl phos phoric acid dimethyl ester as the diazo compound, potassium N OH ---step 1-7 carbonate, potassium t-butoxide and the like can be used as 2 the base. The diazo compound is used in the amount of 1 to 1.5 Hal N R2 equivalents based on compound (1b-6). The base is used in 10 the amount of 1 to 2 equivalents based on compound (1b-6). (1b-7) The reaction temperature is from -78°C. to room tempera O ture, and the reaction time is from 10 minutes to 24 hours. N OH Compound (1b) can also be obtained from compound (1b 15 6) by the methods given below as Alternative Methods (1). R 2 Alternative Method (1): Compound (1b-6) can be con O N R2 Verted into a dihaloalkene in the presence of a base, and then (1b-8) reacted with a base to obtain compound (1b). Dihaloalkene synthesis: There are no particular limitations on the solvent used in this reaction as long as it dissolves the (wherein R and Hal are defined as above, and R represents starting materials to a certain extent without impeding the a C- alkyl.) reaction. Examples of the solvent used in this synthesis Compound (1b-7) which is a commercially available prod include ether solvents such as tetrahydrofuran and diethyl uct can be used as is, or compound (1b-7) can also be manu ether; aromatic hydrocarbon Solvents such as benzene and 25 factured by from a commercially available product with the toluene; and hexane, mixed solvents of the foregoing and the well known methods, for example, WO 2005/033079 A1, pp like. Examples of the reagent for converting compound (1b-6) 85-86, etc. into dihaloalkene include (dichloromethyl)-phosphoric acid Step 1-7 dimethyl ester, dibromomethyl triphenyl phosphonium bro This step is a step wherein compound (1b-8) is obtained by mide (Tetrahedron Letters, Vol. 40, No. 49,8575-8578) and 30 reacting compound (1b-7) with an alcohol in the presence of the like. Examples of the base in this reaction include lithium a base. This step is carried out according to the procedures of diisopropylamide, potassium t-butoxide and the like. The Step 1-39 given below or the method disclosed in Journal of reagent for converting into dihaloalkene is used in the amount Medicinal Chemistry, Vol. 46, No. 5, pp 702-705 or the like. of 1 to 1.5 equivalents based on compound (1b-6). The base is There are no particular limitations on the solvent used in this used in the amount of 1 to 2 equivalents based on compound 35 (1b-6). The reaction temperature is from -78° C. to room reaction as long as it dissolves the starting materials to a temperature, and the reaction time is from 10 minutes to 24 certain extent without impeding the reaction. Examples of the hours. Solvents in this step include ether solvents such as tetrahy As another synthetic method of dihaloalkene, following drofuran and diethyl ether; aromatic hydrocarbon solvents alternative method using carbon tetrabromide can be applied. 40 Such as benzene and toluene; amide solvents such as N.N- Compound (1b-6) is converted into dihaloalkene by reacting dimethylformamide and N-methylpyrrolidinone; alcohol sol carbon tetrabromide and triphenylphosphine. Zinc can also vents such as methanol and ethanol; and dimethyl Sulfoxide, be added in this reaction. There are no particular limitations mixed solvents of the foregoing and the like. Examples of the on the solvent used in this reaction as long as it dissolves the base in this step include Sodium hydride, potassium t-butox starting materials to a certain extent without impeding the 45 ide, potassium hexamethyldisilaZide and the like. A copper reaction. Preferable examples of the solvent used in this syn catalyst can be added in this reaction. Examples of the copper thesis include tetrahydrofuran and methylene chloride. The catalyst include copper, copper(I) iodide, copper(I) bromide, carbon tetrabromide is used in the amount of 1 to 2 equiva copper (I) chloride and the like. The base can be used in the lents based on compound (1b-6). The triphenylphosphine is amount of 1 to 20 equivalents based on compound (1b-7). The used in the amount of 2 to 4 equivalents based on compound 50 alcohol can be used in the amount of 1 to 20 equivalents based (1b-6). The zinc is used in the amount of 1 equivalent based on on compound (1b-7). The copper catalyst can be used in the the carbon tetrabromide. The reaction temperature is from 0° amount of 0.01 to 0.3 equivalents based on compound (1b-7). C. to room temperature, and the reaction time is from 10 The reaction temperature is from 0°C. to reflux temperature, minutes to 12 hours. and the reaction time is from 30 minutes to 48 hours. Synthesis of compound (1b) from dihaloalkene: There are 55 Manufacturing Method 1-2-4 Method 1 for Manufacturing no particular limitations on the Solvent used in this reaction as Compound (1b-4): long as it dissolves the starting materials to a certain extent without impeding the reaction. Examples of the solvent used in this synthesis include ether solvents such as tetrahydrofu R9 ran and diethyl ether, aromatic hydrocarbon solvents such as 60 O No1 Nsna-Bu), benzene and toluene; and hexane, mixed solvents of the fore R7 going and the like. Examples of the base used in this reaction N O1 (1b-9-1) include n-butyl lithium, t-butyl lithium, potassium t-butoxide step1-8) 2 and the like. The base is used in the amount of 2 to 3 equiva Hal N R2 lents based on the dihaloalkene. The reaction temperature is 65 from -78°C. to room temperature, and the reaction time is from 10 minutes to 24 hours. US 8,158,657 B2 25 26 -continued -continued O R7 N O1

O 2 R9 1. N R2 (1b-10) (1b-11) 10 (wherein R, R and Hal are defined as above and R repre (wherein Hal, R and R7 are defined as above, R'' and R' sents a C- alkyl group.) each independently represents C alkyl groups.) Compound (1b-9) which is a commercially available prod Compound (1b-9) and compound (1b-9-2) which are com uct can be used as is, or compound (1b-9) can also be manu 15 mercially available products can be used as is or may be factured from commercially available products by the well obtained from commercially available products by the known known methods. Compound (1b-9-1) which is a commer methods cially available product can be used as is, or compound (1b Step 1-9 9-1) can also be manufactured from commercially available This step is a step wherein compound (1b-11) is obtained products by the well known methods (for example, WO 2005/ by alkylating compound (1b-9) through a reaction with com 033079 A1, pp 85-86, etc.). pound (1b-9-2) in the presence of a palladium catalyst. Com Step 1-8 pound (1b-11) can be manufactured according to the method This step is a step wherein compound (1b-10) is obtained similar to those of Step 1-8. by reacting compound (1b-9) with compound (1b-9-1) in the 25 Manufacturing Mehod 1-2-6. Method for Manufacturing presence of a palladium catalyst. A phosphine ligand may Compound (1b-5) also be added to obtain good results. There are no particular limitations on the solvent used in this reaction as long as it dissolves the starting materials to a certain extent without O impeding the reaction. Examples of the solvents in this step 30 include ether solvents such as 1,4-dioxane and tetrahydrofu N O1 H ran; aromatic hydrocarbon solvents such as toluene and step 1-10 xylene; amide solvents such as N,N-dimethylformamide and 2 N-methylpyrrolidinone; and dimethyl sulfoxide, mixed sol R1 N R2 vents of the foregoing and the like. Examples of the palladium 35 catalyst include palladium (II) acetate, tris(dibenzylideneac (1b-3) etone)dipalladium (O), dichlorobis(triphenylphosphine)pal ladium (II), dichlorobis(tri-o-tolylphosphine)palladium (II), N o1 H bis(tri-t-butylphosphine)palladium (O), tetrakis(triph 40 enylphosphine)palladium (O), 1,1'-bis(diphenylphosphinof 2 errocene)dichloropalladium(II) and the like. Examples of the R1 N R2 phosphine ligand include triphenylphosphine, tri-o- (1b-5) tolylphosphine, tri-t-butylphosphine, diphenylphosphinofer rocene and the like. Compound (1b-9-1) is used in the amount 45 of 1 to 3 equivalents based on compound (1b-9). The palla (wherein R' and Rare defined as above.) dium catalyst is used in the amount of 0.01 to 0.3 equivalents Compound (1b-3) which is a commercially available prod based on compound (1b-9). The phosphine ligand is used in uct can be used as is or may be obtained from commercially the amount of 0.01 to 1.2 equivalents based on compound available products by the known methods. (1b-9). The reaction temperature is from room temperature to 50 Step 1-10 reflux temperature, and the reaction time is from 10 minutes to 24 hours. This step is a step wherein compound (1b-5) is obtained by reducing compound (1b-3). Compound (1b-5) can be manu Manufacturing Method 1-2-5 Method 2 for Manufacturing factured according to methods similar to those of Step 1-4. Compound (1b-4) 55 Manufacturing Method 1-2-7 Method for Manufacturing Halogen-Modified Product of Pyridine Ring R10 I O -SirN R10 60

R12 He N - (1892)step 1-9 step 1-11 2 Hal N 65 (1b-12) US 8,158,657 B2 27 28 -continued -continued N OH Hal N O 2 step 1-13 1. N R2 2 (1b-15) O (1b-13) (wherein R, R and Hal is defined as above; R' represents a 10 N H step 1-14 O 2 hydrogen atom, a hydroxy group, or OR (R is defined as 1. N R2 above).) Compound (1b-12) which is a commercially available (1b-16) O product can used as is or may be obtained from commercially 15 available products by the known methods. Step 1-11 N H This step is a step wherein compound (1b-13) is obtained HO 2 by Substituting a halogen atom for a hydrogen atom on the N R2 pyridine ring of compound (1b-12). This step can be carried out according to, for instance, European Journal of Medicinal (1b-17) Chemistry, Vol. 12, No. 6, 531-536, or, Journal of Organic Chemistry, Vol. 49, No. 26, 5237-5243, or the like. There are (wherein R and R is defined as above.) no particular limitations on the Solvent used in this reaction as Compound (1b-14) can be manufactured according to the 25 methods described in Manufacturing Method 1-2-4 given long as it dissolves the starting materials to a certain extent above. without impeding the reaction. Examples of the Solvent Step 1-12 include halogen solvents such as chloroform and dichlo This step is a step wherein compound (1b-15) is obtained romethane; ether solvents such as tetrahydrofuran and diethyl by reducing compound (1b-14). Compound (1b-15) can be ether; amide solvents such as N,N-dimethylformamide and 30 manufactured according to the methods similar to those of N-methylpyrrolidinone; acid solvents such as acetic acid and Step 1-4. hydrochloric acid aqueous solution; dimethyl sulfoxide; Step 1-13 acetonitrile; mixed solvents of the foregoing, or the like. This step is a step wherein compound (1b-16) is obtained Examples of halogenation reagent include N-chlorosuccin by oxidizing compound (1b-15). Compound (1b-16) can be imide, N-bromosuccinimide, chlorine and bromine. The 35 manufactured according to the methods similar to those of Step 1-5). halogenation reagent is used in the amount of 1.0 to 1.5 Step 1-14 equivalents based on compound (1b-12). The reaction tem This step is a step wherein compound (1b-17) is obtained perature is from room temperature to 50° C., and the reaction by reacting compound (1b-16) with borontribromide. There time is from 5 minutes to 24 hours. are no particular limitations on the solvent used in this reac Manufacturing Method 1-2-8 Method for Manufacturing 40 tion as long as it dissolves the starting materials to a certain Compound (1b-6) extent without impeding the reaction. Examples of the Sol vent include halogenated hydrocarbon Solvents such as meth ylene chloride; aromatic hydrocarbon solvents such as ben Zene and toluene; mixed solvents of the foregoing, or the like. 45 Borontribromide can be used in the amount of 1 to 5 equiva lents based on compound (1b-16), preferably 3 equivalents. step1-12 The reaction temperature is from -20°C. to room tempera ture, and preferably 0°C. The reaction time is 10 minutes to 24 hours. (1b-14) Manufacturing Method 1-3-1 Method 1 for Manufacturing Compound (1 c):

Z Hal Z No NR3 Or NR3

(1c-2) (1c-1) step 1-1 GN splis US 8,158,657 B2 29 30 -continued H 9. Q Horor Z R3 He-step 1-17 or Gr Z YR3 --step-23 o?- N.YG Z YR3 step 1-24 o? N Y-Gr Z NR3 (1c-3) (1c-6) (1c-8) (1c-9) split split (step)-2IN slal step 1-2 GN |cols ? OH N Sa Z n Z N Z HN R - NR3 O? YR3 step1-19 "ror's R 14 HO1 N (1c-5) (1c-4) (1c-7) (1c)

(wherein ring A. R. Zand Hal are defined as above, R' and based on the organometallic compound. The temperature for R" represent Calkyl groups or crosslinked -(CH2), , in reacting the organometallic compound and formylation is 2 or 3, and R'' represents a hydrogenatom, a sodium atom, reagent is from -78°C. to room temperature in the case of the a potassium atom and a lithium atom.) organolithium compounds, with a reaction time being from 5 Each compound in the above reaction scheme which is minutes to 6 hours, while in the case of the Grignard reagents commercially available products can be used as is, or each the reaction temperature is from -78°C. to reflux temperature compound can also be manufactured from commercially 25 of the solvent, with a reaction time being from 5 minutes to 24 available products by the well known methods. In addition, hours. each compound can be manufactured by the methods Step 1-16 described in the manufacturing examples in the examples and This step is a step wherein compound (1c-6) is obtained by by the methods described in Manufacturing Method 1-3-1 30 to Manufacturing Method 1-3-23. reacting an acid to the acetal of compound (1c-2), so as to Step 1-15 deprotect the acetal. There are no particular limitations on the This step is a step wherein compound (1c-6) is obtained by Solvent used in this reaction as long as it dissolves the starting reacting a formylation reagent with an organometallic com materials to a certain extent without impeding the reaction. pound obtained by Substituting a metal atom for the halogen 35 Examples of the solvent used in this reaction include ether atom in compound (1c-1). There are no particular limitations Solvents such as tetrahydrofuran and diethyl ether, aromatic on the solvent used in this reaction as long as it dissolves the hydrocarbon solvents such as benzene and toluene; amide starting materials to a certain extent without impeding the solvents such as N,N-dimethylformamide and N-methylpyr reaction. Preferable examples of the solvents used in this rolidinone; alcohol Solvents such as methanol and ethanol: reaction include ether solvents such as tetrahydrofuran and 40 dimethyl sulfoxide and water, mixed solvents of the forego diethyl ether. Examples of the organometallic compound ing and the like. Examples of the acid in this reaction include include organolithium compounds obtained using a base Such inorganic acids Such as hydrochloric acid, Sulfuric acid, and as n-butyl lithium, s-butyl lithium, t-butyl lithium and lithium hydrobromic acid; organic acids such as citric acid, trifluo diisopropylamide, or Grignard reagents obtained using a base roacetic acid, p-toluenesulfonic acid and the like. The acid Such as metal magnesium, ethyl magnesium bromide and 45 can be used in the amount of from a catalytic amount to an isopropyl magnesium chloride. A catalytic amount of iodine, excess amount based on compound (1c-2). The reaction tem dibromoethane and the like can be added when preparing the perature is from 0°C. to the reflux temperature of the solvent, Grignard reagents using metal magnesium. The temperature and the reaction time is from 5 minutes to 24 hours. for preparing the organolithium compound is from -78°C. to Step 1-17 room temperature, preferably from -78°C. to -40°C., the 50 This step is a step wherein compound (1c-6) is obtained by base is used in the amount of 1 to 1.5 equivalents based on oxidation of compound (1c-3). Compound (1c-6) can be compound (1c-1), and the reaction time is from 30 minutes to 24 hours. The temperature for preparing the Grignard manufactured according to the methods similar to those of reagents using metal magnesium is from room temperature to Step 1-5). reflux temperature of the solvent, the metal magnesium is 55 Step 1-18 used in the amount of 1 to 2 equivalents based on compound This step is a step wherein compound (1c-6) is obtained by (1c-1), and the reaction time is from 30 minutes to 24 hours. reduction of compound (1c-4). Compound (1c-6) can be The temperature for preparing the Grignard reagents using obtained by means of the reduction reaction using a reducing ethyl magnesium bromide or isopropyl magnesium chloride agent Such as diisobutylaluminum hydride, sodium triethoxy is from -60° C. to reflux temperature, the ethyl magnesium 60 aluminum hydride, lithium triethoxyaluminum hydride and bromide or isopropyl magnesium bromide is used in the the like. There are no particular limitations on the solvent amount of 1 to 1.6 equivalents based on compound (1c-1), and used, but in the case of a reducing reaction using a reducing the reaction time is from 5 minutes to 12 hours. Examples of agent, hydrocarbons such as toluene and ethers such as tet the formylation agents include dimethylformamide, rahydrofuran can be used. The reducing agent is used in the N-formylpiperidine, N-formylmorpholine, N-methylforma 65 amount of 1 to 2 equivalents based on compound (1c-4). The nilide and the like. The formylation reagent can be used in the reaction temperature is from 78°C. to room temperature, and amount of 1 to 20 equivalents, preferably 1 to 2 equivalents, the reaction time is from 10 minutes to 24 hours. US 8,158,657 B2 31 32 Step 1-19 rials to a certain extent without impeding the reaction. This step is a step wherein compound (1c-5) is obtained by Examples of the solvents used in this reaction include alcohol reduction of compound (1c-4). Compound (1c-5) can be Solvents such as methanol and ethanol; ether solvents such as obtained either by the reduction reaction using a reducing tetrahydrofuran and diethyl ether; and the like. Examples of agent Such as lithium aluminum hydride or diisobutylalumi the base in this reaction include sodium methoxide, Sodium num hydride, or by catalytic hydrogenation using a Raney ethoxide, n-butyl lithium, lithium diisopropylamide, sodium nickel, palladium-carbon or other catalyst in a hydrogen hydroxide, potassium hydroxide, potassium carbonate, atmosphere. There are no particular limitations on the solvent potassium t-butoxide or the like. The nitromethane can be used, but in the case of a reducing reaction using a reducing used in the amount of 1 to 20 equivalents based on compound agent, ethers such as tetrahydrofuran and diethyl ether, or 10 (1c-6). The base is used in the amount of 1 to 2 equivalents hydrocarbons such as toluene can be used preferably, while in based on compound (1c-6). The reaction temperature is from the case of catalytic hydrogenation, alcohols such as metha -78°C. to reflux temperature, and the reaction time is from 5 nol, ethanol, propanol and the like can be used preferably. The minutes to 48 hours. reducing agent is used in the amount of 1 to 10 equivalents Step 1-22 based on compound (1c-4). There are no particular limita 15 This step is a step wherein compound (1c-8) is obtained by tions on the reaction temperature, but in the case of the reduc esterifying the hydroxyl groups of compound (1c-7) in the ing reaction using a reducing agent, the reaction temperature presence of a base, followed by elimination in situ. There are is from -78°C. to a reflux temperature of the solvent used, no particular limitations on the Solvent used in this reaction as while in the case of the catalytic hydrogenation, the reaction long as it dissolves the starting materials to a certain extent temperature is from room temperature to a reflux temperature without impeding the reaction. Examples of the solvents used of the solventused. The reaction time is from 10 minutes to 24 in this reaction include ether solvents such as tetrahydrofuran hours. The atmospheric pressure in the case of catalytic and diethyl ether; amide solvents such as N,N-dimethylfor hydrogenation is from 1 to 4 atoms. An amount of catalyst mamide and N-methylpyrrolidinone; and methylene chlo from a catalytic amount to excess may be used in catalytic ride, dimethylsulfoxide, mixed solvents of the foregoing and hydrogenation. 25 the like. Examples of the base in this reaction include triethy Step 1-20 lamine, N,N-diisopropylethylamine and the like. Examples This step is a step wherein compound (1c-3) is obtained by of the esterification agents include acetic anhydride, meth converting the amino groups of compound (1c-5) into anesulfonyl chloride, p-toluenesulfonyl chloride and the like. acetoxy groups by reacting with sodium nitrite and acetic The base is used in the amount of 1.0 to 4.0 equivalents based acid, followed by hydrolysis using a base. 30 on compound (1c-7). The esterification agent is used in the Acetoxylation reaction: Preferable example of the solvent amount of 1.0 to 2.0 equivalents based on compound (1c-7). used in this reaction includes a mixed solvent of acetic acid The reaction temperature is from room temperature to reflux and water. More preferably, the ratio of acetic acid to water is temperature, and the reaction time is from 30 minutes to 24 from 1:5 to 5:1. Sodium nitrite is used in the amount of 1 to 20 hours. equivalents based on compound (1c-5). The reaction tem 35 Compound (1c-8) can also be obtained from compound perature is from 0°C. to room temperature, and the reaction (1c-7) by the method described below as Alternative Method time is from 1 hour to 12 hours. (1). Hydrolysis reaction: There are no particular limitations on Alternative Method (1): Compound (1c-8) can be obtained by the solvent used in this reaction as long as it dissolves the dehydrating compound (1c-7) in an acetic acid solvent in the starting materials to a certain extent without impeding the 40 presence of an acetic acid salt. Acetic acid is used as the reaction. Examples of the solvent in this reaction include Solvent in this reaction, but a mixed solvent of acetic acid and alcohol solvents such as methanol and ethanol; ether solvents methanol, tetrahydrofuran and the like can also be used. such as tetrahydrofuran; amide solvents such as N,N-dimeth Examples of acetic acid salt include ammonium acetate, eth ylformamide and N-methylpyrrolidinone; and water, dim ylene diamine diacetic acid salt and the like. The acetic acid ethyl sulfoxide, mixed solvents of the foregoing and the like. 45 salt is used in the amount of 1 to 20 equivalents based on Examples of the base include Sodium hydroxide, potassium compound (1c-7). The reaction temperature is from room hydroxide, potassium carbonate and the like. The reaction temperature to reflux temperature, and the reaction time is temperature is from 0°C. to 60° C., preferably from 20° C. to from 30 minutes to 72 hours. 40°C., and the reaction time is from 30 minutes to 12 hours. Step 1-23 Compound (1c-3) can also be obtained from compound 50 This step is a step wherein compound (1c-8) is obtained by (1c-5) by the method described as Alternative Method (1) reacting compound (1c-6) with nitromethane in the presence below. of a base, and then dehydrating by addition of an acid to the Alternative Method (1): This step is a step whereincompound reaction system. There are no particular limitations on the (1c-3) is obtained by heating compound (1c-5) in the pres Solvent used in this reaction as long as it dissolves the starting ence of a strong base. Preferable example of the solvent in this 55 materials to a certain extent without impeding the reaction. step includes diethylene glycol, and preferable example of the Examples of the solvent used in this reaction include water; base includes potassium hydroxide. The potassium hydrox alcohol solvents such as methanol and ethanol; ether solvents ide is used in the amount of 5 to 30 equivalents based on such as tetrahydrofuran and diethyl ether; mixed solvents of compound (1c-5), the reaction temperature is from 150° C. to the foregoing and the like. Examples of the base used in this 23.0°C., and the reaction time is from 1 hour to 12 hours. Note 60 reaction include sodium methoxide, sodium ethoxide, n-butyl that during the reaction, an inactive gas is preferably Substi lithium, lithium diisopropylamide, Sodium hydroxide, potas tuted inside the reaction container. sium hydroxide, potassium carbonate, potassium t-butoxide Step 1-21 and the like. Examples of the acid used in this reaction include This step is a step wherein compound (1c-7) is obtained by hydrochloric acid, sulfuric acid, acetic acid and the like. The reacting compound (1c-6) with nitromethane in the presence 65 nitromethane is used in the amount of 1 to 20 equivalents of a base. There are no particular limitations on the solvent based on compound (1c-6). The base is used in the amount of used in this reaction as long as it dissolves the starting mate 1 to 2 equivalents based on compound (1c-6). The acid can be US 8,158,657 B2 33 34 added in an excess amount. The reaction temperature for the Step 1-25 reaction with nitromethane is from -78°C. to reflux tempera This step is a step wherein compound (1c) is obtained by an ture, with a reaction time being from 5 minutes to 48 hours. ionization of the nitroethyl moiety in compound (1c-9) using The reaction temperature for the dehydration reaction is from a base, followed by adding titanium (IV) chloride. An ionization reaction of compound (1c-9): There are no room temperature to reflux temperature, with a reaction time particular limitations on the solvent used in this reaction as being from 5 minutes to 48 hours. long as it dissolves the starting materials to a certain extent Compound (1c-8) can also be obtained from compound without impeding the reaction. Examples of the solvents used (1c-6) by the method given below as Alternative Method (1). in this reaction include alcohol Solvents such as methanol and ethanol; ether solvents such as tetrahydrofuran; and the like. Alternative Method (1): Compound (1c-8) can be obtained by 10 Examples of the base used in this reaction include lithium reacting compound (1c-6) with nitromethane in the presence methoxide, Sodium methoxide, potassium t-butoxide, n-butyl of an acetic acid salt. Acetic acid is used as the solvent in this lithium and the like. The base is used in the amount of 1 to 2 reaction, but a mixed solvent of acetic acid and methanol, equivalents based on compound (1c-9). The reaction tem tetrahydrofuran and the like can also be used. Examples of the perature is from -78°C. to room temperature, and the reac acetic acid salt used in this reaction include ammonium 15 tion time is from 5 minutes to 1 hour. acetate, ethylenediamine diacetic acid salt and the like. Reaction with titanium(IV) chloride: There are no particu Nitromethane is used in the amount of 1 to 10 equivalents lar limitations on the solvents used in this reaction as long as it dissolves the starting materials to a certain extent without based on compound (1c-6). The acetic acid salt is used in the impeding the reaction. Examples of the solvents used in this amount of 1 to 20 equivalents based on compound (1c-6). The reaction include ether solvents such as tetrahydrofuran; and reaction temperature is from room temperature to reflux tem methylene chloride, 1,2-dichloroethane, mixed solvents of perature, and the reaction time is from 30 minutes to 72 hours. the foregoing and the like. The titanium (IV) chloride is used Step 1-24 in the amount of 1 to 3 equivalents based on compound (1c-9). This step is a step wherein compound (1c-9) is obtained by The reaction temperature is from -10°C. to room tempera 25 ture, and the reaction time is from 10 minutes to 12 hours. reduction of compound (1c-8). In order to obtain good results, Step 1-26 an acid such as acetic acid or hydrochloric acid can be added. This step is a step wherein compound (1c) is obtained by There are no particular limitations on the solvent used in this reacting compound (1c-8) with titanium (IV) chloride in the reaction as long as it dissolves the starting materials to a presence of triethylsilane. There are no particular limitations certain extent without impeding the reaction. Examples of the on the solvent used in this reaction as long as it dissolves the Solvents used in this reaction include alcohol solvents such as 30 starting materials to a certain extent without impeding the methanol and ethanol; ether solvents such as tetrahydrofuran: reaction. Examples of the solvents used in this reaction and dimethylsulfoxide and the like. Examples of the reducing include ether solvents such as tetrahydrofuran; and methyl ene chloride, 1,2-dichloroethane, mixed solvents of the fore agent used in this reaction include Sodium borohydride, going and the like. The triethylsilane is used in the amount of lithium borohydride and the like. The reducing agent is used 35 1 to 3 equivalents based on compound (1c-8). The titanium in the amount of 0.5 to 3 equivalents based on compound (IV) chloride is used in the amount of 1 to 3 equivalents based (1c-8). The reaction temperature is from -20°C. to 80°C., on compound (1c-8). The reaction temperature is from -20° and the reaction time is from 10 minutes to 12 hours. In the C. to room temperature, and the reaction time is from 10 case of adding the acid, the acid can be added in the amount minutes to 12 hours. of 1 equivalent to the solvent amount based on the reducing Manufacturing Method 1-3-2 Method 2 for Manufacturing agent. Compound (1c)

HO 1n R3 (1c-10-1) O N / step 1-27 N N N n He 1N Hestep 1-29 step 1-30 Ol4\o L R 4N-1- Ol21 1N3 N (1c-10-2) / (1c-10) step 1-28 (1c-11) (1c-12) O ls N N step 1-31 step1-32) R 15 Cl21 1. R3 rC2 1n R3 (1c-13) (1c-14) N N

step 1-33) step 1-34 2 o1N3 4N1\s (1c-15) (1c-16) US 8,158,657 B2 35 36

-continued

(wherein R is defined as above; in the formula, R' repre 10 Step 1-29 sents a C- alkyl group which may be substituted with a This step is a step wherein compound (1c-12) is obtained halogen or the like; L represents a leaving group Such as a by reacting compound (1c-11) with peroxide. Examples of halogen atom, a p-toluenesulfonyl group and a trifluo the peroxide used in this reaction include m-chloroperben romethanesulfonyl group.) Zoic acid, hydrogen peroxide, dimethyldioxirane, benzoyl 15 peroxide, peracetic acid or the like. There are no particular Compound (1c-10), compound (1c-10-1) and compound limitations on the solvent used in this reaction as long as it (1c-10-2) which are commercially available products can be dissolves the starting materials to a certain extent without used as is or they can also be manufactured from commer impeding the reaction. Examples of the solvent include halo cially available products by the known methods. gen solvents such as chloroform and methylene chloride; Step 1-27 alcohol solvents such as methanol and ethanol; amide Sol This step is a step wherein compound (1c-11) is obtained vents such as N,N-dimethylformamide and N-methylpyrro by reacting compound (1c-10) with an organophosphorous lidinone; aromatic hydrocarbon Solvents such as benzene and compound, an azo reagent and compound (1c-10-1). There toluene, diethyl ether, acetone; acetonitrile; acetic acid; water or the like. Peroxide is used in the amount of 1 to 5 equivalents are no particular limitations on the solvent used in this reac based on compound (1c-11). The reaction temperature is tion as long as it dissolves the starting materials to a certain 25 from -40°C. to reflux temperature, and the reaction time is extent without impeding the reaction. Examples of the Sol from 1 minute to 48 hours. vents used in this reaction include ether solvents such as Step 1-30 tetrahydrofuran and diethyl ether; aromatic hydrocarbonsol This step is a step wherein compound (1c-13) is obtained vents such as benzene and toluene; amide solvents such as by reacting compound (1c-12) with an acid anhydride. N,N-dimethylformamide and N-methylpyrrolidinone; ethyl 30 Examples of the acid anhydride used in this reaction include acetate; acetonitrile; methylene chloride; mixed solvents of acetic anhydride, trifluoroacetic acid anhydride, or the like. the foregoing and the like. Examples of the organophospho There are no particular limitations on the solvent used in this rous compound include triphenyl phosphine, tri-n-butyl reaction as long as it dissolves the starting materials to a phosphine and the like. Examples of the azo compound certain extent without impeding the reaction. Examples of the include ester derivatives such as diethylazodicarboxylate and 35 Solvent include halogen solvents such as chloroform and diisopropyl azodicarboxylate, and amide derivatives such as methylene chloride; aromatic hydrocarbon Solvents such as 1,1'-(aZodicarbonyl)dipiperidine. Compound (1c-10-1) is benzene and toluene; acetic acid, trifluoroacetic acid or the used in the amount of 1 to 1.5 equivalents based on compound like. Acid anhydride can also be used as the solvent. Acid (1c-10). The organophosphorous compound is used in the anhydride is used in the amount of 1 equivalent to excess 40 based on compound (1c-12). The reaction temperature is amount of 1 to 3 equivalents based on compound (1c-10). The from 0°C. to reflux temperature, and the reaction time is from aZo reagent is used in the amount of 1 to 3 equivalents based 10 minutes to 24 hours. on compound (1c-10). The reaction temperature is from 0°C. Step 1-31 to reflux temperature, and the reaction time is from 5 minutes This step is a step wherein compound (1c-14) is obtained to 24 hours. 45 by hydrolyzing compound (1c-13). Compound (1c-14) can Step 1-28 be obtained by hydrolyzing compound (1c-13), for instance, This step is a step wherein compound (1c-11) is obtained in the presence of an acid such as Sulfuric acid, or, for by reacting compound (1c-10) with compound (1c-10-2), in instance, in the presence of an alkali Such as sodium hydrox the presence of a base. There are no particular limitations on ide, potassium hydroxide, Sodium methoxide, potassium car the solvent used in this reaction as long as it dissolves the 50 bonate or sodium carbonate. There are no particular limita starting materials to a certain extent without impeding the tions on the solvent used in this reaction as long as it dissolves reaction. Examples of the solvents used in this reaction the starting materials to a certain extent without impeding the include ether solvents such as tetrahydrofuran, diethyl ether reaction. Examples of the solvent include ether solvents such or the like; aromatic hydrocarbon solvents such as benzene, as 1,4-dioxane and tetrahydrofuran; alcohol solvents such as toluene or the like; amide solvents such as N,N-dimethylfor 55 methanol and ethanol; halogen solvents such as methylene mamide, N-methylpyrrolidinone or the like: alcohol solvents chloride and chloroform; aromatic hydrocarbon solvents such as methanol, ethanol or the like; dimethyl sulfoxide: Such as benzene and toluene; amide solvents such as N.N- mixed solvents of the foregoing and the like. Examples of the dimethylformamide and N-methylpyrrolidinone; dimethyl base include Sodium hydride, potassium t-butoxide, sodium Sulfoxide, acetonitrile, water, mixed solvents of the foregoing ethoxide, sodium methoxide, N,N-diisopropylethylamine, 60 or the like. The acid or the base is used in the amount of 1 triethylamine, potassium hydroxide, Sodium hydroxide, equivalent to excess based on compound (1c-13). The reac potassium carbonate, sodium carbonate and the like. The base tion temperature is from 0°C. to reflux temperature, and the is used in the amount of 1 to 5 equivalents based on compound reaction time is from 10 minutes to 24 hours. (1c-10-2). Compound (1c-10-2) is used in the amount of 1 to Step 1-32) 20 equivalents based on compound (1c-10). The reaction 65 This step is a step wherein compound (1c-15) is obtained temperature is from 0° C. to reflux temperature, and the by converting the hydroxyl group of compound (1c-14) to a reaction time is from 5 minutes to 6 hours. leaving group. US 8,158,657 B2 37 38 When L is a methanesulfonyloxy group, p-toluenesulfony compound (1c-14). The reaction temperature is from 0°C. to loxy group or other Sulfuric acid ester, compound (1c-15) can reflux temperature, and the reaction time is from 10 minutes be obtained by reacting compound (1c-14) with sulfonyl to 12 hours. chloride under basic conditions. There are no particular limi Alternative Method (3): Compound (1c-15) can be obtained tations on the solvent used in this reaction as long as it dis by reacting compound (1c-14) with phosphorus halide. There Solves the starting materials to a certain extent without imped are no particular limitations on the solvent used in this reac ing the reaction. Examples of the solvent include ether tion as long as it dissolves the starting materials to a certain Solvents such as 1,4-dioxane and tetrahydrofuran; aromatic extent without impeding the reaction. Examples of the Sol hydrocarbon solvents such as benzene and toluene; amide vent include ether solvents such as diethyl ether; N,N-dim 10 ethylformamide, acetonitrile, chloroform and the like. solvents such as N,N-dimethylformamide and N-methylpyr Examples of the phosphorus halide include phosphorus oxy rolidinone; and dimethyl sulfoxide, methylene chloride, chloride, phosphorus trichloride, phosphorus tribromide and mixed solvents of the foregoing and the like. Examples of the the like. The phosphorus halide is used in the amount of 0.33 base include triethylamine, N,N-diisopropylethylamine and to 3 equivalents based on compound (1c-14). The reaction the like. Examples of the sulfonyl chloride include methane 15 temperature is from 0° C. to reflux temperature, and the sulfonyl chloride, p-toluenesulfonyl chloride and the like. reaction time is from 10 minutes to 12 hours. The base is used in the amount of 1 to 3 equivalents based on Step 1-33 compound (1c-14). The sulfonyl chloride is used in the This step is a step wherein compound (1c-16) is obtained amount of 1 to 2 equivalents based on compound (1c-14). The by converting the leaving group of compound (1c-15) to a reaction temperature is from 0°C. to room temperature, and cyano group. To obtain good results, an inorganic salt such as the reaction time is from 10 minutes to 24 hours. sodium iodide or the like may also be added in the amount of When L is a chlorine atom or a bromine atom; compound 1 to 2 equivalents based on compound (1c-15). Examples of (1c-15) can be obtained by halogenating compound (1c-14) the cyanization agent used in this reaction include sodium with tetrachloromethane or tetrabromomethane in the pres cyanide, potassium cyanide, lithium cyanide or the like. ence of triphenylphosphine. There are no particular limita 25 There are no particular limitations on the solvent used in this tions on the solvent used in this reaction as long as it dissolves reaction as long as it dissolves the starting materials to a the starting materials to a certain extent without impeding the certain extent without impeding the reaction. Examples of the reaction. Examples of the solvent include ether solvents such Solvent include alcohol solvents such as methanol and etha as tetrahydrofuran; amide solvents such as N,N-dimethylfor nol; ether solvents such as 1,4-dioxane and tetrahydrofuran: mamide and N-methylpyrrolidinone; and methylene chlo 30 amide solvents such as N,N-dimethylformamide and N-me thylpyrrolidinone; dimethyl sulfoxide; acetonitrile; acetone; ride, mixed solvents of the foregoing and the like. The tetra water; mixed solvents of the foregoing or the like. The cya chloromethane or tetrabromomethane can also be used as the nidation agent is used in the amount of 1 to 5 equivalents solvent. The triphenylphosphine is used in the amount of 1 to based on compound (1c-15). The reaction temperature is 2 equivalents based on compound (1c-14). The tetrachlo 35 from room temperature to reflux temperature, and the reac romethane or tetrabromomethane is used in the amount of 1 tion time is from 30 minutes to 48 hours. equivalent to the solvent amount based on compound (1c-14). Step 1-34 The reaction temperature is from 0°C. to reflux temperature, This step is a step wherein compound (1c-17) is obtained and the reaction time is from 10 minutes to 12 hours. by reacting compound (1c-16) with hydroxylammonium Compound (1c-15) can also be obtained from compound 40 chloride. Examples of the base used in this reaction include (1c-14) according to the methods described below as Alter pyridine, Sodium acetate, potassium acetate, sodium bicar native Methods (1), (2) and (3). bonate, sodium carbonate, Sodium hydroxide, potassium Alternative Method (1): Compound (1c-14) can be converted hydroxide or the like. There are no particular limitations on into compound (1c-15) under acidic conditions. There are no the solvent used in this reaction as long as it dissolves the particular limitations on the solvent used in this reaction as 45 starting materials to a certain extent without impeding the long as it dissolves the starting materials to a certain extent reaction. Examples of the solvent include halogenated hydro without impeding the reaction. Examples of the Solvent carbons such as dichloromethane and chloroform; Sulfoxides include ether solvents such as diethyl ether; water, ethyl such as dimethyl sulfoxide; ethers such as tetrahydrofuran acetate, mixed solvents of the foregoing and the like. In this and 1,4-dioxane, alcohols such as methanol and ethanol: reaction, a phase-transfer agent such as tetrabutylammonium 50 amides such as N-methylpyrrolidinone, N,N-dimethylforma bromide can be used in the amount of 0.01 to 2 equivalents mide and N,N-dimethylacetamide: pyridine; water, mixed based on compound (1c-14). Examples of the acid include Solvents of the foregoing, or the like. Hydroxylammonium hydrochloric acid, hydrobromic acid and the like. Sulfuric chloride is used in the amount of 1 to 5 equivalents based on acid can also be added to obtain good results. The reaction compound (1c-16). The base is used in the amount of 1 temperature is from 0°C. to room temperature, and the reac 55 equivalent to excess based on compound (1c-16). The reac tion time is from 10 minutes to 12 hours. tion temperature is from 0°C. to reflux temperature, and the Alternative Method (2): Compound (1c-15) can be obtained reaction time is from 10 minutes to 48 hours. by reacting compound (1c-14) with thionyl chloride. There Step 1-35 are no particular limitations on the solvent used in this reac This step is a step wherein compound (1c-18) is obtained tion as long as it dissolves the starting materials to a certain 60 by reacting compound (1c-17) with sodium nitrite and a chlo extent without impeding the reaction. Examples of the Sol rine source. Examples of the chlorine source used in this vent include aromatic hydrocarbon solvents such as benzene reaction include hydrochloric acid, copper chloride or the and toluene; and acetonitrile, chloroform, methylene chloride like. There are no particular limitations on the solvent used in and the like, and the thionyl chloride can also be used as the this reaction as long as it dissolves the starting materials to a solvent. Pyridine can also be added to the reaction in a cata 65 certain extent without impeding the reaction. Examples of the lytic amount to improve the yield. The thionyl chloride is used Solvent include ether solvents such as 1,4-dioxane and tet in the amount of 1 equivalent to the solvent amount based on rahydrofuran; amide solvents such as N,N-dimethylforma US 8,158,657 B2 39 40 mide and N-methylpyrrolidinone; dimethyl sulfoxide; aceto carbonate, sodium carbonate and the like. Examples of the nitrile; acetone; hydrochloric acid aqueous solution; water, copper catalysts include copper, copper(I) iodide, copper(I) mixed solvents of the foregoing, or the like. Sodium nitrite bromide, copper(I) chloride and the like. Compound (1c-20 can be used in the amount of 1 to 10 equivalents based on 1) is used in the amount of 1 to 5 equivalents based on compound (1c-17). The chlorine source can be used in the compound (1c-20). The base is used in the amount of 1 to 5 amount of 1 equivalent to excess based on compound (1c-17). equivalents based on compound (1c-20). The copper catalyst The reaction temperature is from -40°C. to reflux tempera can be used in the amount of 0.01 to 0.3 equivalents based on ture, and the reaction time is from 1 minute to 24 hours. compound (1b-20). The reaction temperature is from 0°C. to Manufacturing Method 1-3-3 Method 1 for Manufacturing reflux temperature, and the reaction time is from 5 minutes to Compound (1c-1): 10 48 hours. Manufacturing Method 1-3-4 Method 2 for Manufacturing Compound (1c-1):

HO-R3 15 B (1c-19-1) r S sess 1N Br HO R3 21 N (1c-10-1) (1c-19) HO-R Br N O-R 2 step 1-39 (1c-19-1) / N Br B step1-37 21 (1c-22) r N pH 1c-21 L-R3 (1c-21) 25 21 N (1c-20-1) / (1c-20) step 1-38 30 o% 1n R3 (1c-23) (wherein R and L are defined as above.) Compound (1c-19), compound (1c-20), compound (1c-19 1) and compound (1c-20-1) which are commercially avail 35 able products can be used as is, or they can be obtained from commercially available products by the well known methods. (wherein R is defined as above.) Step 1-36 Compound (1c-22) and Compound (1c-10-1) may be com This step is a step wherein compound (1c-21) is obtained mercially available products or can also be manufactured by reacting compound (1c-19) with compound (1c-19-1) in 40 from the commercially available products by the well known the presence of a base. Compound (1c-21) can be manufac methods. tured according to the methods similar to those of Step 1-28. Step 1-37 Step 1-39 This step is a step wherein compound (1c-21) is obtained This step is a step wherein compound (1c-23) is obtained by reacting compound (1c-20) with an organophosphorous 45 by reacting compound (1c-22) with compound (1c-10-1) in compound, an azo reagent and compound (1c-19-1). Com the presence of a base. A copper catalyst can also be added in pound (1c-21) can be manufactured according to the methods this reaction. There are no particular limitations on the solvent similar to those of Step 1-27. used in this reaction as long as it dissolves the starting mate Step 1-38 rials to a certain extent without impeding the reaction. This step is a step wherein compound (1c-21) is obtained 50 Examples of the solvents used in this reaction include ether by reacting compound (1c-20) with compound (1c-20-1) in Solvents such as tetrahydrofuran and diethyl ether, aromatic the presence of a base. A catalytic amount of sodium iodide, hydrocarbon solvents such as benzene and toluene; amide potassium iodide or tetrabutylammonium iodide can be solvents such as N,N-dimethylformamide and N-methylpyr added to obtain good results, and a copper catalyst can also be rolidinone; alcohol Solvents such as methanol and ethanol: added in order to obtain good results. There are no particular 55 and dimethyl Sulfoxide, mixed solvents of the foregoing and limitations on the solvent used in this reaction as long as it the like. Examples of the base include sodium hydride, potas dissolves the starting materials to a certain extent without sium t-butoxide, sodium ethoxide, sodium methoxide, potas impeding the reaction. Examples of the solvents used in this sium hydroxide, sodium hydroxide and the like. Examples of reaction include ether solvents such as tetrahydrofuran and the copper catalyst include copper, copper (I) iodide, copper diethyl ether; aromatic hydrocarbon solvents such as benzene 60 (I) bromide, copper(I) chloride and the like. The base is used and toluene; amide solvents such as N,N-dimethylformamide in the amount of 1 to 5 equivalents based on compound and N-methylpyrrolidinone; alcohol solvents such as metha (1c-10-1). Compound (1c-10-1) is used in the amount of 1.0 nol and ethanol; and dimethyl sulfoxide, mixed solvents of to 3.0 equivalents based on compound (1c-22). The copper the foregoing and the like. Examples of the base include catalyst can be used in the amount of 0.01 to 1 equivalents Sodium hydride, potassium t-butoxide, Sodium ethoxide, 65 based on compound (1c-10-1). The reaction temperature is sodium methoxide, N,N-diisopropylethylamine, triethy from room temperature to reflux temperature, and the reac lamine, potassium hydroxide, Sodium hydroxide, potassium tion time is from 10 minutes to 48 hours. US 8,158,657 B2 41 42 Manufacturing Method 1-3-5 Method 3 for Manufacturing desirable. Examples of the organometallic compounds Compound (1c-1), (1c-2) and (1c-6) include organolithium compounds obtained by applying a

Br

(1c-24-1) step1-45

13 13 R a. R a. O O O 13 13 Br N H N R NO N R'sO N - " | ------a step 1-40 a step 1-41 a step 1-42 a H N Hal N Hal N Hal N O (1c-24) (1c-25) (1c-26) (1c-27) HO-R HO-R R 16 step 1-43 step 1-46)1-46 step 1-48 Hal's MI s ^ (1c-19-1) (1c-19-1) step1-49tep1-49 21--R17

(1c-26-1) R13 R13 O NO NO Br R. R 16 R's N H N O N 17. O n % -R step 1-47 % -R % N % HO (1c-31) (1c-32) (1c-33) (1c-28) LR- P3 step 1-44) (1c-20-1) R13No R13 No N

2 O N NR3 (1c-29)

(wherein Hal, L, R, R' and R' is defined as above; in 50 base such as n-butyl lithium, s-butyl lithium, t-butyl lithium addition, R'' and R7 represent a halogen group, a C- alkyl or the like. The temperature at which the organolithium com group and a C- alkoxy group; Hal' represents a chlorine pound is to be prepared is from -100° C. to room temperature, atom and a bromine atom; M' represents a magnesium atom and preferably from -78°C. to -40°C. The base can be used and a Zinc atom.) in the amount of 1 to 1.2 equivalents based on compound Compound (1c-24), compound (1c-25), compound (1c-19- 55 (1c-24), and the reaction time is from 10 minutes to 24 hours. 1), compound (1c-20-1), compound (1c-24-1) and compound Examples of the formylating reagent include N,N-dimethyl (1c-26-1) which are commercially available products can be formamide, N-formylpiperidine, N-formylmorpholine, used as is, or they can be manufactured from commercially N-methylformanilide or the like. The formylating reagent can available products by the known methods. be used in the amount of 1 to 20 equivalents based on com Step 1-40 60 pound (1c-24), and preferably 1 to 2 equivalents. The tem This is a step wherein compound (1c-25) is obtained by perature for reacting the organometallic compound and the converting compound (1c-24) into an organometallic com- formylating reagentis from -78°C. to room temperature, and pound by Substituting a metal atom for a halogen atom of the reaction time is from 5 minutes to 24 hours. compound (1c-24), and then applying a formylating reagent. Step 1-41 There are no particular limitations on the solvent used in this 65 This step is a step wherein compound (1c-26) is obtained step as long as it dissolves the starting materials to a certain by protecting the formyl group of compound (1c-25) with an extent without impeding the reaction, but diethyl ether is acetal, in the presence of an alcohol and an acid catalyst. US 8,158,657 B2 43 44 Preferable examples of alcohol used in this reaction include Step 1-45 methanol, ethanol, ethylene glycol, propylene glycol or the This step is a step wherein compound (1c-30) is obtained like. Examples of the acid catalyst include hydrochloric acid, by reacting compound (1c-24) and compound (1c-24-1), in Sulfuric acid, p-toluenesulfonic acid, acetic acid, ammonium the presence of a base. Compound (1c-30) can be manufac tured according to the methods similar to those of Step 1-39. chloride or the like. There are no particular limitations on the Step 1-46 Solvent used in this step as long as it dissolves the starting This step is a step wherein compound (1c-31) is obtained materials to a certain extent without impeding the reaction. by reacting compound (1c-24) and compound (1c-19-1), in Examples of the solvent include alcohol solvents such as the presence of a base. Compound (1c-31) can be manufac methanol, ethanol and ethylene glycol; aromatic hydrocarbon tured according to the methods similar to those of Step 1-39. Solvents such as benzene and toluene; halogenated hydrocar 10 Step 1-47 bon solvents such as dichloromethane and chloroform, or the This step is a step wherein compound (1c-32) is obtained like. The alcohol is used in the amount of 1 equivalent to the by converting compound (1c-31) into an organometallic com Solvent amount based on compound (1c-25). The acid cata pound by Substituting a metal atom for a halogen atom of compound (1c-31), and then reacting with a formylating lyst is used in the amount of 0.05 equivalents to excess based 15 agent. Compound (1c-32) can be manufactured according to on compound (1c-25). The reaction temperature is from room the methods similar to those of Step 1-15. temperature to reflux temperature, and the reaction time is Step 1-48 from 10 minutes to 24 hours. This step is a step wherein compound (1c-32) is obtained Step 1-42 by reacting compound (1c-25) and compound (1c-19-1), in This step is a step wherein compound (1c-27) is obtained the presence of a base. Compound (1c-32) can be manufac by converting compound (1c-26) into an organometallic com tured according to the methods similar to those of Step 1-39. pound by Substituting a metal atom for a halogen atom of Step 1-49 compound (1c-26), and then applying a formylation reagent. This step is a step wherein compound (1c-33) is obtained Compound (1c-27) can be manufactured according to the by reacting compound (1c-26) and compound (1c-26-1), in methods similar to those of Step 1-15. 25 the presence of a nickel catalyst. There are no particular Step 1-43 limitations on the solvent used in this reaction as long as it This step is a step wherein compound (1c-28) is obtained dissolves the starting materials to a certain extent without by reducing compound (1c-27). Examples of the reducing impeding the reaction. Preferable examples of the solvent agent used in this reaction include Sodium borohydride, include ether solvents such as tetrahydrofuran, dioxane and lithium borohydride, lithium aluminum hydride, or the like. 30 diethyl ether. Examples of the nickel catalyst include 1,3-bis There are no particular limitations on the solvent used in this (diphenylphosphino)propane nickel (II) chloride, bis(triph reaction as long as it dissolves the starting materials to a enylphosphine) nickel (II) chloride, 1.2-bis(diphenylphos certain extent without impeding the reaction. Examples of the phino)ethane nickel (II) chloride, 1,1'-bis Solvent include alcohol solvents such as methanol and etha (diphenylphosphino) ferrocene nickel(II) chloride or the like. nol; ether solvents such as diethyl ether and tetrahydrofuran: 35 Compound (1c-26-1) is used in the amount of 1 to 2 equiva aromatic hydrocarbon Solvents such as benzene and toluene; lents based on compound (1c-26), and the nickel catalyst is halogenated hydrocarbon Solvents such as dichloromethane used in the amount of 0.02 to 0.2 equivalents based on com and chloroform; water, mixed solvents of the foregoing, or the pound (1c-26). The reaction temperature is from -10°C. to like. Preferably alcohol solvents are desirable when using a 80°C., and the reaction time is from 30 minutes to 12 hours. reducing agent Such as Sodium borohydride; ether solvents 40 In addition, when M' is a zinc atom, compound (1c-26-1) are desirable when using a reducing agent such as lithium can be prepared in situ and used in the reaction, as describe aluminumhydride. The reducing agent is used in the amount below. The reaction in Step 1-49 can be carried out by of 0.25 to 4 equivalents based on compound (1c-27). The synthesizing compound (1c-26-1) in situ using benzyl halide reaction temperature is from 0°C. to reflux temperature, and and activated zinc. In this case, the activated Zinc is used in the the reaction time is from 5 minutes to 24 hours. 45 amount of 1 to 1.3 equivalents based on benzyl halide. The Step 1-44 reaction temperature for obtaining compound (1c-26-1) is This step is a step wherein compound (1c-29) is obtained from -10°C. to room temperature, preferably from -5°C. to by reacting compound (1c-28) and compound (1c-20-1), in 10°C., and the reaction time is from 1 hour to 10 hours. the presence of a base. Compound (1c-29) can be obtained Manufacturing Method 1-3-6. Method 1 for Manufacturing according to the methods similar to those of Step 1-38. Compound (1c-2)

US 8,158,657 B2 47 48 (wherein R. R. R. and R' represent a halogen, a Cs Manufacturing Method 1-3-7 Method 2 for Manufacturing alkyl group and a C- alkoxy group.) Compound (1c-2) Compound (1c-34), compound (1c-34-1), compound (1c 34-2), compound (1c-34-3) and compound (1c-34-4) which Hal R22 are commercially available products can be used as is, or they N can also be manufactured from commercially available prod | X,- R23 ucts by the known methods. le Step 1-50 This step is a step wherein compound (1c-35) is obtained (1c-37-1) by converting compound (1c-34) into an organometallic com step1-54) pound by Substituting a metal atom for a halogen atom of compound (1c-34), and then reacting with compound (1c-34 (1C-37)

1). There are no particular limitations on the solvent used in 15 this reaction as long as it dissolves the starting materials to a certain extent without impeding the reaction. Preferable examples of the solvent include ether solvents such as tet rahydrofuran and diethyl ether. Examples of the organome tallic compound include organolithium compounds obtained by applying a base Such as n-butyl lithium, S-butyl lithium, (1C-38) t-butyl lithium, lithium diisopropyl amide, or the like. The temperature at which the organolithium compound is to be (wherein Hal is defined as above; R’ and R represent a prepared is from -78° C. to room temperature, preferably halogen atom, a C- alkyl group and a C- alkoxy group.) 25 Compound (1c-37) and compound (1c-37-1) which are from -78°C. to -40°C. The base is used in the amount of 1 commercially available products can be used as is, or they can to 1.5 equivalents based on compound (1c-34). The reaction be manufactured from commercially available products by time is from 30 minutes to 24 hours. Compound (1c-34-1) is the known methods. used in the amount of 1 to 2 equivalents based on compound Step 1-54) (1c-34). The temperature for reacting the organometallic 30 This step is a step wherein compound (1c-38) is obtained by converting compound (1c-37) into an organometallic com compound and compound (1c-34-1) is from -78°C. to room pound by Substituting a metal atom for the hydrogen atom at temperature, and the reaction time is from 5 minutes to 12 position 5 of compound (1c-37), and then reacting with com hours. pound (1c-37-1). There are no particular limitations on the Step 1-51 35 Solvent used in this reaction as long as it dissolves the starting This step is a step wherein compound (1c-35) is obtained materials to a certain extent without impeding the reaction. by reacting compound (1c-34) and compound (1c-34-2), in Preferable examples of the solvent include ether solvents such as tetrahydrofuran and diethyl ether. Examples of the the presence of a base. There are no particular limitations on organometallic compound include organolithium compounds the solvent used in this reaction as long as it dissolves the 40 obtained by applying a base such as n-butyl lithium, S-butyl starting materials to a certain extent without impeding the lithium, t-butyl lithium, lithium diisopropylamide or the like. reaction. Examples of the solvent include ether solvents such The temperature at which the organolithium compound is to as tetrahydrofuran and diethyl ether; aromatic hydrocarbon be prepared is from -78°C. to room temperature, preferably Solvents such as benzene and toluene; amide solvents such as from -78°C. to -40°C. The base is used in the amount of 1 N,N-dimethylformamide and N-methylpyrrolidinone; alco 45 to 1.5 equivalents based on compound (1c-37). The reaction hol solvents such as methanol and ethanol; dimethyl sulfox time is from 30 minutes to 24 hours. Compound (1c-37-1) is ide, mixed solvents of the forgoing, or the like. Examples of used in the amount of 1 to 2 equivalents based on compound the base include sodium hydride, potassium t-butoxide, (1c-37). The temperature for reacting the organometallic Sodium ethoxide, sodium methoxide, potassium hydroxide, compound and compound (1c-37-1) is from -78°C. to room sodium hydroxide or the like. The base is used in the amount 50 temperature. The reaction time is from 5 minutes to 12 hours. of 1 to 5 equivalents based on compound (1c-34). Compound Manufacturing Method 1-3-8 Method 1 for Manufacturing (1c-34-2) is used in the amount of 1 to 2 equivalents based on Compound (1c-3): compound (1c-34). The reaction temperature is from room temperature to reflux temperature, and the reaction time is L-R3 from 5 minutes to 24 hours. 55 Step 1-52 This step is a step wherein compound (1c-36) is obtained rs2 /" (1c-20-1)step 1-55 by converting compound (1c-34) into an organometallic com pound by Substituting a metal atom for a halogen atom of (1c-39) compound (1c-34), and then reacting compound (1c-34-3). 60 Compound (1c-36) can be manufactured according to the HO Ny methods similar to those of Step 1-50. Step 1-53 21 This step is a step wherein compound (1c-36) is obtained (1c-40) by reacting compound (1c-34) and compound (1c-34-4) in 65 the presence of a base. Compound (1c-36) can be manufac tured according to the methods similar to those of Step 1-51. (wherein Rand L are defined as above.) US 8,158,657 B2 49 50 Compound (1c-39) and Compound (1c-20-1) may be com compound is from -78° C. to reflux temperature, with a mercially available products or can also be manufactured reaction time being from 10 minutes to 12 hours. The reaction from the commercially available products by the well known temperature in the reaction of adding compound (1c-41-1) is methods. from -78°C. to room temperature, with a reaction time being Step 1-55 from 10 minutes to 6 hours. This step is a step wherein compound (1c-40) is obtained Step 1-57 by reacting compound (1c-39) with compound (1c-20-1). This step is a step wherein compound (1c-43) is obtained Compound (1c-20-1) is used in the amount of 0.2 to 1.0 by deprotecting the acetal of compound (1c-42). Compound equivalents based on compound (1c-39). Compound (1c-40) (1c-43) is manufactured according to the methods similar to can be manufactured according to the methods similar to 10 those of Step 1-16). those of Step 1-38. Step 1-58 Manufacturing Method 1-3-9 Method 2 for Manufacturing This step is a step wherein compound (1c-43) is obtained Compound (1c-3): by reacting compound (1c-41) with compound (1c-41-1). In

R13 R13 No OHC-R3 No R13 (1c-41-1) R13 3 No N step 1-56 No S R --Hall 3-( 2 21 OH (1c-41) (1c-42) NOHC-R3 |cols,

O

H N R3 HO N R3 3-( step 1-59 / 2 OH 21 (1c-43) (1c-44)

(wherein R. R. R'' and Hal are defined as above.) this step, compound (1c-42) is obtained according to the Compound (1c-41) and compound (1c-41-1) may be com methods similar to those of Step 1-56, after which an acid is mercially available products or can also be manufactured added in the reaction system or at the work-up stage to obtain from the commercially available products by the well known 45 compound (1c-43). Examples of the acids used in this reac methods. tion include inorganic acids such as hydrochloric acid, Sulfu Step 1-56 ric acid and hydrobromic acid; organic acids such as citric This step is a step wherein the halogenatom in compound acid, trifluoroacetic acid and p-toluenesulfonic acid; acidic (1e-41) is replaced with a metal atom to obtain an organome silica gels and the like. The acid can be used in the amounts of tallic compound, which is then reacted with compound (1c 50 from the catalytic amount to the solvent amount based on 41-1) to obtain compound (1c-42). There are no particular Compound (1c-41). The reaction temperature is from 0°C. to limitations on the solvent used in this reaction as long as it the reflux temperature of the solvent, and the reaction time is dissolves the starting materials to a certain extent without from 5 minutes to 24 hours. impeding the reaction. Examples of the solvents used in this Step 1-59 reaction include ether solvents such as tetrahydrofuran and 55 diethyl ether; aromatic hydrocarbon solvents such as benzene This step is a step wherein compound (1c-44) is obtained and toluene; mixed solvents of the foregoing and the like. by reducing compound (1c-43). There are no particular limi Examples of the reagent for converting compound (1c-41) tations on the solvent used in this reaction as long as it dis into an organometallic compound include n-butyl lithium, Solves the starting materials to a certain extent without imped S-butyl lithium, ethyl magnesium bromide, ethyl magnesium 60 ing the reaction. Examples of the solvents used in this chloride, isopropyl magnesium chloride, magnesium, Zinc reaction include ether solvents such as tetrahydrofuran and and the like. The reagent for converting compound (1c-41) diethyl ether; aromatic hydrocarbon solvents such as benzene into an organometallic compound is used in the amount of 1 and toluene; and the like. Examples of the reducing agent to 3 equivalents based on compound (1c-41). Compound include lithium aluminum hydride-aluminum chloride and (1c-41-1) is used in the amount of 1 to 2 equivalents based on 65 the like. The lithium aluminum hydride is used in the amount compound (1c-41). The reaction temperature in the reaction of 2 to 6 equivalents based on compound (1c-43). The alumi for converting compound (1c-41) into an organometallic num chloride is used in the amount of 2 to 9 equivalents based US 8,158,657 B2 51 52 on compound (1c-43). The reaction temperature is from 0°C. Manufacturing Method 1-3-11 Method 4 for Manufacturing to reflux temperature, and the reaction time is from 10 min Compound (1c-3) utes to 48 hours. Manufacturing Method 1-3-10 ethod 3 for Manufacturing Compound (1c-3) O R. No Her Hal step 1-62 Hal I N N 10 R24 o R24 (1c-48) O 2 21 O RQ7 OHC-R No N OH (1c-45-1) O (1c-41-1) step1-60 15 -- P step 1-63 2 N O (1c-45) R251 R25

2O (1c-49) O R O No -- R24 step1-64 O No N Her 25 21 R3 step 1-61 2 (1c-50) N O (1c-46) R 30 No Her step1-65

R3 R24 O (1c-51) HO N 35 HO 2 N R3 (1c-47) (1c-52) 40 (wherein Hal is defined as above; R represents a hydrogen (wherein R. R7 and Hal are defined as above; in addition, R atom, a halogen, a C- alkyl group and a C- alkoxy group. represents a C- alkyl group.) or the like.) Compound (1c-48) and compound (1c-41-1) which are Compound (1c-45) and compound (1c-45-1) which are 45 commercially available products can be used as is, or they can commercially available products can be used as is or they can be manufactured from commercially products by the known also be manufactured from commercially available products methods. by the known methods. Step 1-62 Step 1-60 This step is a step wherein compound (1c-49) is obtained This step is a step wherein compound (1c-46) is obtained 50 by Substituting a phosphorus atom for a halogen atom of by reacting compound (1c-45) and compound (1c-45-1). compound (1c-48). This reaction is carried out by mixing Compound (1c-46) can be obtained by reacting compound compound (1c-48) and trialkylphosphite in a solvent or in the (1c-45) and compound (1c-45-1), for instance, in a solvent absence of solvent, and heating. There are no particular limi such as tetrahydrofuran, N,N-dimethylformamide or dim tations on the solvent used in this reaction as long as it dis ethylsulfoxide, for instance, in the presence of a base such as 55 Solves the starting materials to a certain extent without imped potassium t-butoxide. Compound (1c-45-1) is used in the ing the reaction. Examples of the solvent include aromatic amount of 1 to 1.5 equivalents based on compound (1c-45). hydrocarbon solvents such as toluene and Xylene or mixed The base is used in the amount of 1 to 1.5 equivalents based on solvents of the foregoing. The trialkylphosphite is added in compound (1c-45). The reaction temperature is from room the amount of 1 to 1.2 equivalents based on compound (1c temperature to reflux temperature. The reaction time is from 60 48). The reaction temperature is from 100° C. to 150°C., and 30 minutes to 24 hours. the reaction time is from 30 minutes to 2 hours. Step 1-63 Step 1-61 This step is a step wherein compound (1c-50) is obtained This step is a step wherein compound (1c-47) is obtained by adding a base to compound (1c-49) and then reacting with by reducing the compound (1c-46). Compound (1c-47) can 65 compound (1c-41-1). There are no particular limitations on be manufactured according to the methods similar to those of the solvent used in this reaction as long as it dissolves the Step 1-4. starting materials to a certain extent without impeding the US 8,158,657 B2 53 54 reaction. Examples of the solvent include ether solvents such -continued as tetrahydrofuran and 1,4-dioxane, amide solvents such as O N,N-dimethylformamide and N-methylpyrrolidinone; or mixed solvents of the foregoing. Examples of the base include HO metal hydrides Such as sodium hydride and potassium R26 Her hydride; metal alcoholates such as Sodium methoxide and S aSA step1-67 potassium t-butoxide. Compound (1c-41-1) is added in the so R27 amounts of 1 to 2 equivalents based on compound (1c-49). 2 The reaction temperature is from room temperature to 80°C., and the reaction time is from 30 minutes to 12 hours. 10 (1C-54) Step 1-64 HO 26 This step is a step wherein a double bond of compound S sas R (1c-50) is hydrogenated, leading to compound (1c-51). This s step is a reaction whereby hydrogen addition is carried with 15 o R27 compound (1c-50) in a solvent, under a hydrogen atmo 21 sphere, and using a metal catalyst. There are no particular limitations on the solvent used in this reaction as long as it (1C-55) dissolves the starting materials to a certain extent without impeding the reaction. Examples of the Solvent include ether (wherein Rand R7 represent a halogen atom, a C- alkyl Solvents such as tetrahydrofuran and 1,4-dioxane, alcohol group and a Calkoxy group.) Solvents such as methanol and ethanol; ester Solvents such as Compound (1c-53) and compound (1c-53-1) which are ethyl acetate or mixed solvents of the foregoing. Examples of commercially available products can be used as is, or they the metal catalyst include palladium (II) oxide, palladium 25 may be manufactured from commercially available products hydroxide, platinum (IV) oxide, Raney nickel or the like. The by the known methods. metal catalyst is used in a catalytic amount to excess based on Step 1-66 compound (1c-50). The reaction temperature is from room This step is a step wherein compound (1c-54) is obtained temperature to 80°C., the reaction time is from 5 minutes to by reacting compound (1c-53) and compound (1c-53-1). 24 hours. The reaction pressure is from 1 atmosphere to 4 30 There are no particular limitations on the solvent used in this atmospheres. reaction as long as it dissolves the starting materials to a Step 1-65 certain extent without impeding the reaction. Examples of the Solvent include alcohol solvents such as ethanol and metha This step is a step wherein alcohol product (1c-52) is nol, or the like. Compound (1c-53-1) is used in the amount of obtained by reducing the ester group of compound (1c-51). 35 1 to 2 equivalents based on compound (1c-53). The reaction There are no particular limitations on the solvent used in this temperature is the reflux temperature, and the reaction time is reaction as long as it dissolves the starting materials to a from 30 minutes to 12 hours. certain extent without impeding the reaction. Examples of the Step 1-67 solvent include ether solvents such as ether and tetrahydro This step is a step wherein compound (1c-55) is obtained furan; aromatic hydrocarbon solvents such as toluene and 40 Xylene; or mixed solvents of the foregoing. Examples of the by reducing compound (1c-54). Compound (1c-55) can be reducing agent include sodium borohydride, lithium alumi manufactured according to the methods similar to those of num hydride, diisobutylaluminum hydride, or the like. The Step 1-4. reducing agent is added in the amount of 0.5 to 2 equivalents Manufacturing Method 1-3-13 Method 1 for Manufacturing of compound (1c-51). The reaction temperature is from -20° 45 Compound (1c-4): C. to reflux temperature of the solvent, and the reaction time is from 10 minutes to 24 hours. N S HO-R3 N S. Manufacturing Method 1-3-12 Method 5 for Manufacturing n N (1c-19-1) n N , Compound (1c-3) 50 - Hal step1-68 -HO 21 2 (1c-56) (1c-57) N HO-R N nS. nS. R26 55 N OH (1e-19-1) S. On Na" S. Sé S / step1-69) S / SR --R27 21 21 O 21 (1c-58) (1c-59) 60 (1C-53-1) N HO-R3 HO step1-66 & Br S. Hal (1c-19-1) / - / step 1-70 21 (1C-53) 65 US 8,158,657 B2 55 56 -continued Step 1-70 N HN-R N This step is a step wherein compound (1c-59) is obtained Šs Šs R3 by reacting compound (1c-60) with compound (1c-19-1). N (1c-56-1) S / Compound (1c-59) can be obtained according to the methods - HHal step 1-71 N similar to those of Step 1-36. 2 2 Step 1-71 (1c-56) (1c-61) This step is a step wherein compound (1c-61) is obtained HN-R3 by reacting compound (1c-56) with compound (1c-56-1) in s 2 the presence of a palladium catalyst. A phosphine ligand can S. 9 (1c-56-1) 10 also be added to the reaction system to obtain good results. } { step 1-72 There are no particular limitations on the solvent used in this 21 H reaction as long as it dissolves the starting materials to a (1c-62) certain extent without impeding the reaction. Examples of the Solvent in this reaction include ether solvents such as 1,4- N 15 N N 3 dioxane and tetrahydrofuran; alcohol solvents such as metha / R nol and ethanol; aromatic hydrocarbons solvent Such as tolu ene and xylene; amide solvents such as N.N- 2 dimethylformamide and N-methylpyrrolidinone; and (1c-63) dimethyl sulfoxide, mixed solvents of the foregoing and the s OHC-R- P3 like. Examples of palladium catalyst include palladium (II) acetate, tris(dibenzylidenacetone)dipalladium (O), dichloro N (1c-41-1) bis(triphenylphosphine)palladium (II), dichlorobis(tri-o- --NH step 1-73 tolylphosphine)palladium (II), bis(tri-t-butylphosphine)pal 2 25 ladium (O), tetrakis(triphenylphosphine)palladium (O), palladium (O) pentadienone and the like. Examples of the (1c-64) phosphine ligand include triphenylphosphine, tri-o- NŠs tolylphosphine, tri-t-butylphosphine, diphenylphosphinofer rocene, 2-dicyclohexylphosphinobiphenyl, 2-di-t-butylphos S^e phinobiphenyl, 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl 2 (BINAP) and the like. Examples of the base include sodium t-butoxide, cesium carbonate, potassium carbonate, potas (1c-65) sium phosphate and the like. Compound (1c-56-1) is used in the amount of 1 equivalent to excess based on compound (wherein R and Hal are defined as above.) 35 (1c-56). The palladium catalyst is used in the amount of 0.01 Compound (1c-56), compound (1c-58), compound (1c to 0.3 equivalents based on compound (1c-56). The phos 60), compound (1c-62), compound (1c-64), compound (1c phine ligand is used in the amount of 0.01 to 1.2 equivalents 19-1), compound (1c-41-1) and compound (1c-56-1) which based on compound (1c-56). The base is used in the amount are commercially available products can be used as is, or they of 1 to 4 equivalents based on compound (1c-56). The reac can also be manufactured from the commercially available 40 tion temperature is from room temperature to reflux tempera products by the well known methods. ture, and the reaction time is from 30 minutes to 72 hours. Step 1-68 Step 1-72 This step is a step wherein compound (1c-57) is obtained This step is a step wherein compound (1c-63) is obtained by reacting compound (1c-56) with compound (1c-19-1) in by a reductive amination in which compound (1c-62) is the presence of a base. There are no particular limitations on 45 reacted with compound (1c-56-1). Acetic acid can also be the solvent used in this reaction as long as it dissolves the added to promote the reaction. There are no particular limi starting materials to a certain extent without impeding the tations on the solvent used in this reaction as long as it dis reaction. Examples of the solvent used in this reaction include Solves the starting materials to a certain extent without imped ether solvents such as tetrahydrofuran; amide solvents such as ing the reaction. Examples of the solvent in this reaction N,N-dimethylformamide and N-methylpyrrolidinone; alco 50 include ether solvents such as 1,4-dioxane and tetrahydrofu hol Solvents such as methanol and ethanol; and dimethyl ran; alcohol solvents such as methanol and ethanol; and meth Sulfoxide, mixed solvents of the foregoing and the like. ylene chloride, mixed solvents of the foregoing an the like. Examples of the base include Sodium hydride, potassium Examples of the reducing agent include lithium aluminum t-butoxide, sodium ethoxide, sodium methoxide, N,N-diiso hydride, sodium borohydride, sodium cyanoborohydride, propylethylamine, triethylamine, potassium hydroxide, 55 sodium triacetoxyborohydride, 2-picoline-borane and the Sodium hydroxide, potassium carbonate, Sodium carbonate like. Compound (1c-56-1) is used in the amount of 1 to 2 and the like. The base is used in the amount of 1 to 5 equiva equivalents based on compound (1c-62). The reducing agent lents based on compound (1c-19-1). Compound (1c-19-1) is is used in the amount of 0.5 to 2 equivalents based on com used in the amount of 1 equivalent to the solvent amount pound (1c-62). The reaction temperature is from room tem based on compound (1c-56). The reaction temperature is 60 perature to reflux temperature, and the reaction time is from from room temperature to reflux temperature, and the reac 10 minutes to 24 hours. tion time is from 30 minutes to 48 hours. Step 1-73 Step 1-69 This step is a step wherein compound (1c-65) is obtained This step is a step wherein compound (1c-59) is obtained by a reductive amination in which compound (1c-64) is by reacting compound (1c-58) with compound (1c-19-1). 65 reacted with compound (1c-41-1). Compound (1c-65) can be Compound (1c-59) can be manufactured according to the manufactured according to the methods similar to those of methods similar to those of Step 1-37. Step 1-72). US 8,158,657 B2 57 58 Manufacturing Method 1-3-14 Method 2 for Manufacturing -continued Compound (1c-4): N OH Šs Q \ f R step 1-77) N& OHC-R N (1c-41-1) (1c-69) -- Hal step 1-74 2 10 (1c-56) \ | * N S (1c-70) n N R3

3-( step 1-75 15 (wherein R is defined as above, and Q represents a sulfur 21 OH atom and oxygen atom.) Compound (1c-68) and compound (1c-41-1) which are (1c-66) commercially available products can be used as is, or they can N S also be manufactured from the commercially available prod n N R3 ucts by the well known methods. Step 1-76 21 This step is a step wherein one of the bromide atoms in compound (1c-68) is an ionized using an organometallic (1c-67) reagent, which is reacted with compound (1c-41-1), then the 25 other bromide atom in compound (1c-68) is an ionized by adding the further organometallic reagent in the same con (wherein R and Hal are defined as above.) tainer, which is then reacted with a cyanization reagent to Compound (1c-56) and compound (1c-41-1) which are obtain compound (1c-69). There are no particular limitations commercially available products can be used as is, or com on the solvent used in this reaction as long as it dissolves the pound (1c-56) and compound (1c-41-1) can also be manufac 30 starting materials to a certain extent without impeding the tured from commercially available products by the well reaction. Examples of the solvent include ether solvents such known methods. as tetrahydrofuran and diethyl ether; aromatic hydrocarbon Solvents such as benzene and toluene; mixed solvents of the Step 1-74 foregoing and the like. Examples of the organometallic This step is a step wherein compound (1c-66) is obtained 35 reagent include n-butyl lithium, s-butyl lithium and the like. by reacting compound (1c-56) with compound (1c-41-1). Preferable examples of the cyanization reagent include Compound (1c-66) can be manufactured according to the p-toluenesulfonyl cyanide. The organometallic reagent is methods similar to those of Step 1-56. used in the total amount of 2 to 3 equivalents based on com Step 1-75 pound (1c-68). Compound (1c-41-1) is used in the amount of 40 1 to 1.5 equivalents based on compound (1c-68). The cya This step is a step wherein compound (1c-67) is obtained nization reagent is used in the amount of 1 to 1.5 equivalents by reducing compound (1c-66) with iodotrimethylsilane. based on compound (1c-68). The reaction temperature is There are no particular limitations on the solvent used in this from -78°C. to room temperature, and the reaction time is reaction as long as it dissolves the starting materials to a from 10 minutes to 24 hours. certain extent without impeding the reaction. Examples of the 45 Step 1-77 solvent include ether solvents such as tetrahydrofuran; and This step is a step wherein compound (1c-70) is obtained acetonitrile, methylene chloride and the like, preferably by reducing compound (1c-69). Compound (1c-70) can be methylene chloride and acetonitrile. The iodotrimethylsilane manufactured according to the methods similar to those of is used in the amount of 2 to 10 equivalents based on com Step 1-75. pound (1c-66), the reaction temperature is from 0°C. to 60° 50 Manufacturing Method 1-3-16 Method 1 for Manufacturing C., and the reaction time is from 5 minutes to 6 hours. The Compound (1c-5): iodotrimethylsilane used in the reaction may be a commer cially available product, or may be prepared at the time of use by reacting sodium iodide and chlorotrimethylsilane inaceto N nitrile at room temperature. 55 N-a Manufacturing Method 1-3-15 Method 3 for Manufacturing 3-( step 1-78) Compound (1c-4): 21 OH (1c-66) 60 OHC-R HN Br Q (1c-41-1) \ f step 1-76 65 (1c-68) (wherein R is defined as above.) US 8,158,657 B2 59 60 Compound (1c-66) can be manufactured from a commer Compound (1c-42) can be manufactured from commer cially available product by the well known methods, or com cially available products by the well known methods, or com pound (1c-66) can be manufactured according to the methods pound (1c-42) can be manufactured according to the methods similar to those of Step 1-74. similar to those of Step 1-56. Step 1-78 Step 1-80 This step is a step wherein compound (1c-71) is obtained This step is a step wherein compound (1c-73) is obtained by reducing compound (1c-66). There are no particular limi by simultaneous reduction and acetal deprotection of com tations on the solvent used in this reaction as long as it dis pound (1c-42) using iodotrimethylsilane. There are no par Solves the starting materials to a certain extent without imped ticular limitations on the solvent used in this reaction as long ing the reaction. Examples of the solvent include ether 10 as it dissolves the starting materials to a certain extent without Solvents such as tetrahydrofuran and diethyl ether, aromatic impeding the reaction. Examples of the Solvent include ether hydrocarbon Solvents such as benzene and toluene and the Solvents such as tetrahydrofuran; acetonitrile, methylene like. Examples of the reducing agent include lithium alumi chloride and the like, and preferably methylene chloride or num hydride-aluminum chloride. The lithium aluminum 15 acetonitrile. The iodotrimethylsilane is used in the amount of hydride is used in the amount of 3 to 8 equivalents based on 2 to 10 equivalents based on compound (1c-42). The reaction compound (1c-66). The aluminum chloride is used in the temperature is from 0°C. to 60°C., and the reaction time is amount of 3 to 10 equivalents based on compound (1c-66). from 5 minutes to 6 hours. The iodotrimethylsilane used in the The reaction temperature is from 0°C. to reflux temperature, reaction may be a commercial product, or may be prepared at and the reaction time is from 10 minutes to 48 hours. the time of use by reacting sodium iodide and chlorotrimeth Manufacturing Method 1-3-17 Method 2 of Manufacturing ylsilane in acetonitrile at room temperature. Compound (1c-5): Manufacturing Method 1-3-19 Method 2 for Manufacturing Compound (1c-6)

N OH 25 Šs Q 3 \ f R step 1-79 rs--R28 O -N42 (1c-69) 30 (1c-74-1) Q 3 H step 1-81)

orrs OH (1c-72) (1c-74) 35 O (wherein R and Q are defined as above.) Compound (1c-69) can be manufactured from commer H 21 cially available products by the well known methods, or com Sn --Rs pound (1c-69) can be manufactured according to the methods 40 O - similar to those of Step 1-76). Step 1-79 (1c-75) This step is a step wherein compound (1c-72) is obtained by reducing compound (1c-69). Compound (1c-72) can be (wherein R represent a halogen or a C- alkyl group.) manufactured according to the methods similar to those of 45 Compound (1c-74) and compound (1c-74-1) which are Step 1-78). commercially available products can be used as is, or they Manufacturing Method 1-3-18 Method 1 for Manufacturing may be manufactured from commercially available products Compound (1c-6): by the known methods. Step 1-81) 50 This step is a step wherein compound (1c-75) is obtained R13 by reacting compound (1c-74) and compound (1c-74-1), in No the presence of a base. There are no particular limitations on R13 the solvent used in this reaction as long as it dissolves the No N step 1-80 starting materials to a certain extent without impeding the 55 reaction. Examples of the solvent include ether solvents such 2 OH as tetrahydrofuran and 1,4-dioxane; aromatic hydrocarbon Solvents such as benzene and toluene; amide solvents such as (1c-42) N,N-dimethylformamide and N-methylpyrrolidinone, dim O ethyl sulfoxide, mixed solvents of the foregoing, or the like. 60 Examples of the base include Sodium hydride, potassium H N / t-butoxide, Sodium hydroxide, potassium hydroxide, Sodium bicarbonate, potassium carbonate or the like. Compound (1c 21 74) is used in the amount of 0.5 to 2 equivalents based on (1c-73) compound (1c-74-1). The base is used in the amount of 0.5 to 65 5 equivalents based on compound (1c-74-1). The reaction temperature is from 100° C. to 170° C., and the reaction time (wherein R, R and R'' are defined as above.) is from 30 minutes to 12 hours. US 8,158,657 B2 61 62 Manufacturing Method 1-3-20 Method 3 for Manufacturing Compound (1c-6) A. NN O

R29 HO Sé--R30 step1-84 21 (1c-76-1) R29 HO s

O C.21 R30 O 29 N& S O Y S O R (1c-76-1) H SA Hestep1-82 step 1-83 \ f -R 21 21 (1c-77) (1c-78)

(wherein RandR' represent a halogen, a C- alkyl group Manufacturing Method 1-3-21Method 4 for Manufacturing and a C- alkoxy group.) 30 Compound (1c-6) Compound (1c-76), compound (1c-79) and compound (1c 76-1) which are commercially available products can be used as is, or they may be manufactured from commercially avail HN able products by the known methods. 35 Sé--R32 Step 1-82 O 2 This step is a step wherein compound (1c-77) is obtained O by reacting compound (1c-76) and compound (1c-76-1), in H N (1c-80-1) step 1-85 the presence of a bas. There are no particular limitations on O the solvent used in this reaction as long as it dissolves the 40 starting materials to a certain extent without impeding the - O reaction. Examples of the solvent include ether solvents such (1c-80) as tetrahydrofuran and 1,4-dioxane; aromatic hydrocarbon Solvents such as benzene and toluene; amide solvents such as O N,N-dimethylformamide and N-methylpyrrolidinone; dim 45 ethyl sulfoxide, mixed solvents of the foregoing, or the like. Examples of the base include Sodium hydride, potassium Y N R31 t-butoxide, Sodium hydroxide, potassium hydroxide, sodium -/ DYS- R32 bicarbonate, potassium carbonate, or the like. Compound 50 le (1c-76-1) is used in the amount of 1 to 2 equivalents based on compound (1c-76). The base is used in the amount of 2 to 3 (1c-81) equivalents based on compound (1c-76). The reaction tem perature is from room temperature to 80°C., and the reaction (wherein R and R represent a halogen, a C- alkyl group time is from 30 minutes to 72 hours. 55 and a C- alkoxy group.) Compound (1c-80) and compound (1c-80-1) which are Step 1-83 commercially available products can be used as is, or they This step is a step wherein compound (1c-78) is obtained may be manufactured from commercially available products by reducing the cyano group of compound (1c-77). Com by the known methods. pound (1c-78) can be manufactured according to the methods 60 Step 1-85 similar to those of Step 1-18. This step is a step wherein compound (1c-81) is obtained Step 1-84 T by reacting compound (1c-80) and compound (1c-80-1). Examples of the solvent include acetic acid, or the like. Com his step is a step wherein compound (1c-78) is obtained by pound (1c-80-1) is used in the amount of 1 equivalent based reacting compound (1c-79) and compound (1c-76-1), in the 65 on compound (1c-80). The reaction temperature is from 50° presence of a base. Compound (1c-78) can be obtained C. to 110° C., and the reaction time is from 5 minutes to 1 according to the methods similar to those of Step 1-82. hours. US 8,158,657 B2 63 64 Manufacturing Method 1-3-22 Method 5 for Manufacturing (wherein ring A and Hal are defined as above; R represents Compound (1c-6) a C-alkyl group, a Cs cycloalkyl group, a Co aryl group. or a 5- or 6-membered ring heteroaryl group, which may have 1 or 2 substituents selected from the substituent group C.", respectively; M' represents a potassium cation and a sodium cation. ( HN-R Substituent Group C.' O (1c-56-1) a cyano group, a C- alkyl group, a C- alkoxy group, a ---step 1-86) Coalkoxycarbonyl group and a C-8 cycloalkyl group) Compound (1c-85), compound (1c-85-1), compound (1c 85-2) and compound (1c-85-3) which are commercially O available products can be used as is, or they may also be manufactured from commercially available products by the (1c-82) known methods. Step 1-88 ( 15 This step is a step wherein compound (1c-86) is obtained O --- by reacting compound (1c-85) and compound (1c-85-1) or step 1-87 compound (1c-85-2), in the presence of a palladium catalyst and a base. An inorganic salt such as lithium chloride; an NR3 ammonium salt Such as tetrabutylammonium chloride; or a phosphine ligand can also be added to obtain good results. (1c-83) This reaction can be carried out under an inert gas atmo O sphere. Such as nitrogen gas and argon gas. There are no particular limitations on the solvent used in this reaction as H long as it dissolves the starting materials to a certain extent 25 without impeding the reaction. Examples of the Solvent NR3 include ether solvents such as 1,4-dioxane and tetrahydrofu ran; aromatic hydrocarbon solvents such as toluene and (1c-84) xylene; amide solvents such as N,N-dimethylformamide and N-methylpyrrolidinone; dimethyl sulfoxide, water, mixed Solvents of the foregoing, and the like. Examples of the pal (wherein R is defined as above.) 30 ladium catalyst include palladium (II) acetate, tetrakis(triph Compound (1c-82) and compound (1c-56-1) which are enylphosphine)palladium (O), dichlorobis(triphenylphos commercially available products can be used as is, or they phine)palladium(II), tris(dilbenzylidene acetone)dipalladium may be manufactured from commercially available products (O), palladium carbon, bis(tri-t-butyl phosphine)palladium by the known methods. (O), 1,1'-bis(diphenyl phosphinoferrocene)dichloro palla Step 1-86 35 dium (II), or the like. Examples of the phosphine ligand This step is a step wherein compound (1c-83) is obtained include triphenylphosphine, tri-o-tolylphosphine, tri-t-bu by carrying out a reductive amination of compound (1c-82) tylphosphine, tricyclohexyl phosphine, diphenylphosphinof and compound (1c-56-1). Compound (1c-83) can be manu errocene, 2-dicyclohexylphosphino-2,6'-dimethoxybiphe factured according to the methods similar to those of Step nyl, 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl, 1-72). 40 2-di-t-butylphosphino-2',4',6'-triisopropylbiphenyl, 2-di-t- Step 1-87 butylphosphinobiphenyl, 2-dicyclohexylphosphinobiphenyl, This step is a step wherein compound (1c-84) is obtained 2-dicyclohexylphosphino-2'-(N,N-dimethylamino)biphenyl, by deprotecting of acetal in compound (1c-83) by adding an 2-di-t-butylphosphino-2'-(N,N-dimethylamino)biphenyl, acid. Compound (1c-84) can be manufactured according to 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl, 1.2-bis(diphe the methods similar to those of Step 1-16. 45 nylphosphino)ethane, 1,3-bis(diphenylphosphino)propane, Manufacturing Method 1-3-23 Method 6 for Manufacturing 1,4-bis(diphenylphosphino)butane, or the like. Examples of Compound (1c-6) the base include potassium carbonate, Sodium carbonate, cesium carbonate, potassium fluoride, cesium fluoride, potas sium phosphate, sodium hydroxide, barium hydroxide, potas sium hydroxide, or the like. Compound (1c-85-1) or com OH F 50 pound (1c-85-2) is used in the amount of 1 to 3 equivalents R33-B or R33-B-F based on compound (1c-85). The palladium catalyst is used in M+ OH F the amount of 0.01 to 0.25 equivalents based on compound (1c-85). The phosphine ligand is used in the amount of 0.01 to (1c-85-1) (1c-85-2) 1 equivalent based on compound (1c-85). Inorganic salts such step 1-88 55 as lithium chloride, or ammonium salts such as tetrabutylam H monium chloride, are used in the amount of 0.5 to 2 equiva Hal R33-Sn(n-Bu) lents. The reaction temperature is from room temperature to O (1c-85-3) reflux temperature, and the reaction time is from 10 minutes (1c-85) step 1-89 to 72 hours. 60 Step 1-89 H This step is a step wherein compound (1c-86) is obtained R33 by reacting compound (1c-85) with compound (1c-85-3). under a palladium catalyst. An inorganic salt Such as lithium O chloride, an ammonium salt such as tetrabutylammonium (1c-86) 65 chloride, a phosphine ligand, or a copper reagent can also be added to obtain good results. This reaction can be carried out under an inert gas atmosphere. Such as nitrogen gas and argon US 8,158,657 B2 65 66 gas. There are no particular limitations on the solvent used in the addition of an acid, and finally by reacting with a diol Such this reaction as long as it dissolves the starting materials to a as pinacol. This reaction can also be carried out by adding an certain extent without impeding the reaction. Examples of the organometallic reagent to a mixture of compound (1c-87) and Solvent include ether solvents such as 1,4-dioxane and tet boric acid ester, in which the generation of an ion from com rahydrofuran; aromatic hydrocarbon Solvents such as toluene 5 pound (1c-87) and reacting with boric acid ester are occurred and xylene; amide solvents such as N,N-dimethyl formamide simultaneously. This reaction can also be carried out under an and N-methyl pyrrolidinone; dimethyl sulfoxide, mixed sol inert gas stream or atmosphere, such as nitrogen or argon. vents of the foregoing, or the like. Examples of the palladium Examples of compound (1c-87) include chloroiodomethane, catalyst include palladium (II) acetate, tris(dibenzylideneac dibromo methane, bromoiodomethane, or the like, and pref etone)dipalladium (O), dichlorobis(triphenylphosphine)pal- 10 erably, chloroiodomethane and dibromo methane. There are ladium (II), dichlorobis(tri-o-tolylphosphine)palladium (II), no particular limitations on the Solvent used in this reaction as bis(tri-t-butylphosphine)palladium (O), tetrakis(triph- long as it dissolves the starting materials to a certain extent enylphosphine)palladium (O), 1,1'-bis(diphenylphosphinof- without impeding the reaction. Examples of the Solvent errocene)dichloro palladium (II), or the like. Examples of the include ether solvents such as tetrahydrofuran, 1.2- phosphine ligand include triphenylphosphine, tri-o- 15 dimethoxyethane, methyl-t-butyl ether, cyclopentyl methyl tolylphosphine, tri-t-butylphosphine, diphenylphosphinofer- ether, diethyl ether, diisopropyl ether, dibutyl ether and dicy rocene, or the like. Examples of the copper reagent include clopentyl ether, aromatic hydrocarbon Solvents such as ben copper (I) iodide, copper (I) bromide, copper (I) chloride, or Zene and toluene; aliphatic hydrocarbon solvents such as the like. Compound (1c-85-3) is used in the amount of 1 to 3 heptane and hexane, mixed solvents of the foregoing, or the equivalents based on compound (1c-85). The palladium cata- 20 like, and preferably tetrahydrofuran. Examples of the boric lyst is used in the amount of 0.01 to 0.25 equivalents based on acid ester include trimethylborate, triisopropylborate or the compound (1c-85). The phosphine ligand is used in the like, and preferably triisopropylborate. Examples of the orga amount of 0.01 to 1 equivalent based on compound (1c-85). nometallic reagent include n-butyl lithium, s-butyl lithium The copper reagent is used in the amount of 0.1 to 3 equiva- and the like, and preferably n-butyl lithium. Examples of the lents based on compound (1c-85). Inorganic salt Such as 25 acid include methanesulfonic acid, p-toluenesulfonic acid, lithium chloride, or ammonium salt Such as tetrabutylammo- hydrochloric acid-ethyl acetate solution, hydrochloric acid nium chloride, are used in the amount of 0.5 to 2 equivalents. methanol solution, or the like, and preferably methane The reaction temperature is from room temperature to reflux Sulfonic acid and hydrochloric acid-ethyl acetate solution. temperature, and the reaction time is from 10 minutes to 72 Boric acid ester can be used in the amount of 0.8 to hours. 30 1.2 equivalents based on compound (1c-87), and preferably Manufacturing Method 1-3-24 Method for Manufacturing 0.9 to 1 equivalents. The organometallic reagent can be used Compound (1c-85-2) in the amount of 0.8 to 1.2 equivalents based on compound

Hal R34-O \-Hi \-sa-Bu (1c-87) (1c-91) ision solo R3'-OH Hal \-5/ -- R3-O\-i?-F / - R3-OV M step1-91 V M+ step 1-92 CH3 O F (1c-88) (1c-89) (1c-90)

(wherein Hal and Hal" each independently represents a halo- 50 (1c-87), and preferably 0.8 to 1 equivalents. A mixture of the genatom; R represents a Calkyl group, a CalkoxyC. an ionized compound prepared at -78° C. from compound alkyl group, a Css cycloalkyl C- alkyl group or a Cas (1c-87) and boric acid ester is stirred for 1 to 3 hours at the cycloalkyl group; M' represents a potassium cation and a temperature mentioned below. This mixture is neutralized at Sodium cation.) the temperature mentioned below, then pinacol is added and Compound (1c-87), compound (1c-88-1), compound (1e- 55 stirred for 10 to 60 minutes at the temperature mentioned 90) and compound (1c-91) which are commercially available below. products can be used as is, or they may be manufactured from commercially available products by the known methods. |Reaction Temperature During the Reaction of Anionized Compound (1c-91) which a commercially available product Compound and Boric Acid Ester can be used as is, or may be manufactured from commercially 60 The mixture of an ionized compound and boric acid ester is available products by the known methods (for instance, stirred at 0°C. to room temperature, and more preferably at WO2005/033079 A1, pages 82-84, and the like). room temperature. Step 1-90 This step is a step wherein compound (1c-88) is manufac Reaction Temperature During Neutralization Reaction and tured by reacting an organometallic reagent and compound 65 Reaction with Diol (1c-87) to generate an ionized compound, which is reacted The temperature during the neutralization reaction and the with boric acid ester, then neutralizing the reaction mixture by addition of the diol is from -20°C. to room temperature, and US 8,158,657 B2 67 68 more preferably 0°C. The temperature after the addition of Solvent or using a large amount of compound (1c-90) as the the diol is from 0°C. to room temperature, and more prefer Solvent. In addition, this step can be carried out in the pres ably from room temperature ence of a base. This step can be carried out according to the Step 1-91 general methods, for instance, 5" Edition of Jikkenka This step is a step whereinanionized compound, generated 5 gakukoza 18 (pages 20 to 23), Tetrahedron Letters, Vol. 24. by reacting compound (1c-88-1) with a base, is reacted with No. 31, pp. 3165-31 68, and the like. This reaction can also be compound (1c-88), followed by reacting with a hydrogen carried out under an inert gas stream or atmosphere. Such as fluoride salt (potassium hydrogen fluoride or sodium hydro nitrogen or argon. There are no particular limitations on the gen fluoride) to obtain compound (1c-89). This step can also Solvent used in this reaction as long as it dissolves the starting be carried out by adding a catalytic amount of iodine com 10 materials to a certain extent without impeding the reaction. pound Such as potassium iodide and tetrabutylammonium Examples of the solvent include aliphatic hydrocarbon sol iodide. This reaction can also be carried out under an inert gas vents such as heptane and hexane, or the like. Preferably, stream or atmosphere. Such as nitrogen or argon. There are no compound (1c-90) is used in large amount as a solvent. particular limitations on the solvent used in this reaction as Examples of the organometallic reagent include t-butyl long as it dissolves the starting materials to a certain extent 15 lithium, sec-butyl lithium, or the like, preferably sec-butyl without impeding the reaction. Examples of the Solvent lithium. Examples of the base include potassium t-butoxide, include ether solvents such as tetrahydrofuran, 1.2- potassium sec-butoxide, potassium methoxide, or the like, dimethoxyethane, methyl-t-butyl ether, cyclopentyl methyl preferably potassium t-butoxide. An organometallic reagent ether, diethyl ether, diisopropyl ether, dibutyl ether and dicy is added to a mixture of compound (1c-90) and solvent at -75 clopentyl ether; aromatic hydrocarbon Solvents such as ben to -60° C. (preferably -75 to -70° C.), then the mixture is Zene and toluene; amide solvents such as N,N-dimethylfor stirred for 5 to 30 minutes (preferably 5 to 10 minutes) at -20 mamide and N-methylpyrrolidinone; dimethyl sulfoxide, to 0°C. (preferably -10 to -5°C.). Then, boric acid ester is mixed solvents of the foregoing, or the like, and preferably added to the mixture at -75 to -70° C., the mixture is then tetrahydrofuran or N,N-dimethylformamide. Examples of stirred for 10 to 60 minutes (preferably 10 to 30 minutes) at the base include sodium hydride, potassium bis(trimethylsi 25 10° C. to room temperature (preferably room temperature). lyl)amide, potassium hydride, and preferably sodium hydride Hydrogen fluoride salt is added to the mixture at 0 to 5°C., and potassium bis(trimethylsilyl)amide. Compound (1c-88 water is then added at the same temperature, the reaction 1) can be used in the amount of 1 to 5 equivalents based on mixture is warmed to room temperature, to obtain compound compound (1c-88), and preferably 2 to 3 equivalents. The (1c-89). Preferably, compound (1c-90) is used in solvent above-mentioned base can be used in the amount of 1 to 5 30 amount based on the organometallic reagent. The base can be equivalents based on compound (1c-88), and preferably 2 to used in the amount of preferably 0.6 to 1 equivalents based on 3 equivalents. The above-mentioned hydrogen fluoride salt the organometallic reagent. The boric acid ester can be used in can be used in the amount of 2 to 8 equivalents based on the amount of 1 to 2 equivalents based on the organometallic compound (1c-88), and preferably 3 to 5 equivalents. reagent, and preferably 1 to 1.8 equivalents. The above-men The reaction time for aniozation reaction of compound 35 tioned hydrogen fluoride salt can be used in the amount of 3 (1c-88-1) is, preferably from 30 to 60 minutes for stirring for to 10 equivalents based on the above-mentioned boric acid at the temperature mentioned below, and after adding com ester compound, and preferably 3 to 5 equivalents. pound (1c-88) to the mixture, the reaction time is from 1 to 12 Step 1-93 hours for stirring at the temperature mentioned below. After This step is a step wherein anionized compound, generated adding hydrogen fluoride salt to the reaction mixture, the 40 by reactinga organometallic reagent with compound (1c-91), reaction time is from 10 to 120 minutes for stirring at the reacted with a boric acid ester (triisopropylborate, trimethyl temperature mentioned below. borate, 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaboro Reaction Temperature During an ionization Reaction lane, or the like), followed by reacting with a hydrogen fluo The temperature during addition of the base is from 0°C. to ride salt (potassium hydrogen fluoride or Sodium hydrogen room temperature, more preferably 0°C. The temperature 45 fluoride, or the like) to obtain compound (1c-89). This reac after addition of the base is from 0° C. to 70° C., more tion can also be carried out under an inert gas stream or preferably from room temperature to 50° C. atmosphere. Such as nitrogen or argon. There are no particular Reaction Temperature During Reaction of Anionized Com limitations on the solvent used in this reaction as long as it pound and Compound (1c-88) dissolves the starting materials to a certain extent without The temperature during addition of compound (1c-88) is 50 impeding the reaction. Examples of the Solvent include ether from 0° C. to room temperature, and more preferably 0°C. Solvents such as tetrahydrofuran, 1.2-dimethoxyethane, The temperature after addition of compound (1c-88) is from methyl t-butyl ether, cyclopentyl methyl ether, diethyl ether, room temperature to 100° C., more preferably from room diisopropyl ether, dibutyl ether and dicyclopentyl ether; aro temperature to 70° C. matic hydrocarbon Solvents such as benzene and toluene; Reaction Temperature During Addition of Hydrogen Fluo 55 aliphatic hydrocarbon Solvents such as heptane and hexane, ride Salt mixed solvents of the foregoing, and the like, and preferably The temperature during addition of the reagent is from 0° tetrahydrofuran. Examples of the above-mentioned organo C. to room temperature, and more preferably 0°C. The tem metallic reagent include n-butyl lithium, Sec-butyl lithium, perature after addition of the reagent is from 0°C. to room methyl lithium or the like, preferably n-butyl lithium. Com temperature, and more preferably room temperature. 60 pound (1c-89) can be obtained by the two methods described Step 1-92 below. If it is difficult to carry out the reaction (i), such as the This step is a step wherein an ionized compound generated anion generated by reacting organometallic reagent and com by reacting an organometallic reagent and compound (1c-90) pound (1c-91) is unstable, reaction (ii) is preferred. is reacted with boric acid ester, followed by reacting with a (i) In a solvent, an organometallic reagent and compound hydrogen fluoride Salt (potassium hydrogen fluoride or 65 (1c-91) are stirred for 30 to 120 minutes (preferably 30 to 60 Sodium hydrogen fluoride, or the like) to obtain compound minutes) at -75 to -60° C. (preferably -75 to -70° C.). Then, (1c-89). In this step, the reaction can be carried out in a boric acid ester is added to the mixture at -75 to -70° C., US 8,158,657 B2 69 70 whereafter the mixture is stirred for 10 to 120 minutes (pref manufactured according to the methods described in the erably 20 to 80 minutes) at 0°C. to room temperature (pref Manufacturing Examples in the Examples, Manufacturing erably 0 to 5° C.). A hydrogen fluoride salt is added to the Method 2-2-1 given below or the like. mixture at 0 to 5° C., water is then added at the same tem Compound (2c) which is a commercially available product perature, and the reaction mixture is warmed to room tem- 5 can be used as is, or compound (2c) can also be manufactured perature, to obtain compound (1c-89). from commercially available products by the well known (ii) In a solvent, an organometallic reagent is added to a methods. Compound (2c) can further be manufactured mixture of aboric acid ester and compound (1c-89) at -75 to according to the methods described in the Manufacturing -60° C. (preferably -75 to -70° C.), and stirred for 10 to 120 Examples in the Examples, Manufacturing Method 2-3 minutes (preferably 20 to 60 minutes) at -75 to 5°C. (pref 10 given below or the like. erably 0 to 5° C.). Hydrogen fluoride salt is added to the Step 2 mixture at 0 to 5° C., whereafter water is added at the same This step is a step wherein compound (2a) is manufactured temperature, and the reaction mixture is warmed to room by reacting compound (2b) and compound (2c) in the pres temperature, to obtain compound (1c-89). ence of a base. There are no particular limitations on the The organometallic reagent can be used in the amount of 15 Solvent used in this reaction as long as it dissolves the starting 0.8 to 1.2 equivalents based on compound (1c-91), and pref materials to a certain extent without impeding the reaction. erably 1 equivalent. The boric acid ester can be used in the amount of 1 to 2 equivalents based on compound (1c-91), and Examples of the solvent include ether solvents such as tet preferably 1 to 1.2 equivalents. The hydrogen fluoride salt can rahydrofuran and diethyl ether; aromatic hydrocarbon sol be used in the amount of 3 to 10 equivalents based on com vents such as benzene and toluene; amide solvents such as pound (1c-91), and preferably 3 to 5 equivalents. N,N-dimethylformamide and N-methylpyrrolidinone; alco Manufacturing Method 2 Typical Method for Manufactur hol Solvents such as methanol and ethanol; and dimethyl ing Compound (2a): Sulfoxide, mixed solvents of the foregoing and like. Examples of the base include sodium hydride, potassium t-butoxide, 25 Sodium ethoxide, triethylamine, sodium hydroxide, potas (2a) sium hydroxide and the like. Compound (2c) is used in the Z amount of 1 to 5 equivalents based on compound (2b). The ( )- NR3 base is used in the amount of 1 to 5 equivalents based on N compound (2b). The reaction temperature is from 0° C. to Y 30 reflux temperature, and the reaction time is from 10 minutes R M to 24 hours. Manufacturing Method 2-1-2 Method 2 for Manufacturing Compound (2a): r 2 R1 N R2 35 (wherein ring A. R. R. R. R. and Z are defined as above.) Manufacturing Method 2-1-1 Method 1 for Manufacturing Compound (2a): 40

H N R MN + N 45 N 2 (2d) RI N R2 (2b) C ^ns Z N L YR3 - - 50 step2 N N M (2c) ^ns 2 / ( ) 55 RI N R2 N (2a-1) R MN (wherein ring A. R. R. R. Hal and Z are defined as above.) r2 60 Compound (1b-1) which is a commercially available prod RI N R2 uct can be used as is, or compound (1b-1) can also be manu factured from commercially available products by the well (2a) known methods. Compound (2d) can be manufactured from commercially available products by the well known methods, (wherein ring A. R', R. R. Rand Z are defined as above.) 65 or can be manufactured according to the methods described in Compound (2b) can be manufactured from commercially the Manufacturing Examples in the Examples, Manufactur available products by the well known methods, or can also be ing Method 2-4 given below or the like. US 8,158,657 B2 71 72 Step 2-1 ladium (O), tetrakis(triphenylphosphine)palladium (O) and This step is a step wherein compound (2a-1) is obtained by the like. Examples of the phosphine ligand include triph reacting compound (1b-1) with compound (2d) in the pres enylphosphine, tri-o-tolylphosphine, tri-t-butylphosphine ence of a palladium catalyst. Inorganic salts such as lithium and the like. Examples of the copper reagent include copper (I) iodide, copper(I) bromide, copper(I) chloride and the like. chloride, ammonium salts such as tetrabutylammonium chlo Compound (2d) is used in the amount of 1 to 3 equivalents ride, phosphine ligands, or copper reagents can be added to based on compound (1b-1). The palladium catalyst is used in obtain good results. There are no particular limitations on the the amount of 0.01 to 0.3 equivalents based on compound Solvent used in this reaction as long as it dissolves the starting (1b-1). The phosphine ligand is used in the amount of 0.01 to materials to a certain extent without impeding the reaction. 10 1.2 equivalents based on compound (1b-1). The copper Examples of the solvent include ether solvents such as 1,4- reagent is used in the amount of 0.1 to 3 equivalents based on dioxane and tetrahydrofuran; aromatic hydrocarbon solvents compound (1b-1). The inorganic salts such as lithium chlo Such as toluene and Xylene, amide solvents such as N.N- ride or ammonium salts such as tetrabutylammonium chlo dimethylformamide and N-methylpyrrolidinone; dimethyl ride are used in the amount of 1 to 3 equivalents based on Sulfoxide, mixed solvents of the foregoing and the like. 15 compound (1b-1). The reaction temperature is from room Examples of the palladium catalyst include palladium (II) temperature to reflux temperature, and the reaction time is acetate, tris(dibenzylidenacetone)dipalladium (O), dichloro from 10 minutes to 48 hours. bis(triphenylphosphine)palladium (II), dichlorobis(tri-o- Manufacturing Method 2-1-3 Method 3 for Manufacturing tolylphosphine)palladium (II), bis(tri-t-butylphosphine)pal Compound (2a)

OH F R33-B O R-i?-f Hal OH Mt. 7-O- (1c-85-1) (1c-85-2) N N (2c-1) \ step2-3 A N step2-2 M R38-Sn(n-Bu) (1c-85-3) r 2 r2 R2 step2-4 RI N R2 RI N (2b-1) (2a-2)

45 (whereinring A, Hal, L, R', RandR are defined as above.) a base. Compound (2a-3) can be manufactured according to Compound (2b-1) can be manufactured from commer the methods similar to those of Step 1-88. cially available products by the known methods, and can also Step 2-4 be manufactured according to the methods described in the Manufacturing Example in Examples or Manufacturing 50 This step is a step wherein compound (2a-3) is obtained by Method 2-2-1, and the like. Compound (2c-1) which is a reacting compound (1c-85-3) and compound (2a-2), in the commercially available product can be used as is, or may also presence of a palladium catalyst. Compound (2a-3) can be be manufactured from commercially available products by manufactured according to the methods similar to those of the known methods. Compounds (1c-85-1), (1c-85-2) and Step 1-89. (1c-85-3) which are commercially available products can be 55 Manufacturing Method 2-2-1 Method 1 for Manufacturing used as is, or may also be manufactured from commercially Compound (2b): available products by the known methods. Step 2-2 This step is a step wherein compound (2a-2) is obtained by 60 reacting compound (2b-1) and compound (2c-1), in the pres H N ence of a base. Compound (2a-2) can be manufactured M according to the methods similar to those of Step 2. M N -e-step2-5 Step 2-3 Br This step is a step wherein compound (2a-3) is obtained by 65 reacting compound (1c-85-1) or compound (1c-85-2) and (2b-2) compound (2a-2), in the presence of a palladium catalyst and US 8,158,657 B2 73 74 -continued hydroxide and the like. The base is used in the amount of 1 to 4 equivalents based on compound (2b-2). The chlorotriph enylmethane is used in the amount of 1 to 4 equivalents based on compound (2b-2). The reaction temperature is from room O temperature to reflux temperature, and the reaction time is from 1 hour to 24 hours. step2-6 Step 2-6 N O This step is a step wherein compound (2b-4) is obtained by MN reacting compound (2b-3) with a boronic acid derivative in Br 10 the presence of a palladium catalyst and a base. A phosphine ligand may also be added to obtain good results. There are no (2b–3) particular limitations on the solvent used in this reaction as long as it dissolves the starting materials to a certain extent without impeding the reaction. Examples of the Solvent O r" 15 include ether solvents such as 1,4-dioxane and tetrahydrofu 2 ran; aromatic hydrocarbon solvents such as benzene and tolu ene; amide solvents such as N,N-dimethylformamide and N O R1 (1b-1)N R2 N-methylpyrrolidinone; and dimethyl sulfoxide, mixed sol vents of the foregoing and the like. Examples of the palladium O/ step2-7 catalyst include palladium (II) acetate, tris(dibenzylidenac ONB etone)dipalladium (O), dichlorobis(triphenylphosphine)pal ladium (II), bis(tri-t-butylphosphine)palladium (O), tetrakis O (triphenylphosphine)palladium (0), 1,1'-bis (diphenylphosphinoferrocene)dichloropalladium (II) and the 25 like, preferably 1,1'-bis(diphenylphosphinoferrocene)dichlo (2b-4) ropalladium (II). Examples of the base include potassium acetate, triethylamine, N,N-diisopropylethylamine, potas sium phenoxide, potassium carbonate and the like, preferably potassium acetate. Examples of the boronic acid derivative 30 include bis(pinacolate)diboron, 4.4.5.5,-tetramethyl-1,3,2- dioxaborolane and the like. Examples of the phosphine ligand include triphenylphosphine, tri-t-butylphosphine, tricyclo N step2-8 hexylphosphine, diphenylphosphinoferrocene, 2-dicyclo N / hexylphosphinobiphenyl and the like. The palladium catalyst 35 is used in the amount of 0.01 to 0.3 equivalents based on compound (2b-3). The base is used in the amount of 1 to 10 r 2 equivalents based on compound (2b-3). The boronic acid RI N R2 derivative is used in the amount of 1 to 3.0 equivalents based (2b-5) on compound (2b-3). The phosphine ligand is used in the H 40 amount of 0.01 to 1.2 equivalents based on compound (2b-3). N The reaction temperature is from room temperature to reflux MN temperature, and the reaction time is from 10 minutes to 24 hours. r Compound (2b-4) can also be obtained from compound 2 45 (2b-3) according to the method given below as Alternative RI N R2 Method (1). (2b-1) Alternative Method (1): Compound (2b-4) can be obtained by first an ionizing the bromine atom of compound (2b-3) using an organometallic reagent, and then reacting with a boronic (wherein R, R and Hal are defined as above.) 50 acid ester. There are no particular limitations on the solvent A commercially available product can be used as is for used in this reaction as long as it dissolves the starting mate compound (2b-2). Compound (1b-1) which is a commer rials to a certain extent without impeding the reaction. cially available product can be used as is or compound (1b-1) Examples of the solvent include ether solvents such as tet can also be manufactured from commercially available prod rahydrofuran and diethyl ether; aromatic hydrocarbon sol ucts by the well known methods. 55 vents such as benzene and toluene; and hexane, mixed sol Step 2-5 vents of the foregoing and the like. Examples of the This step is a step wherein compound (2b-3) is obtained by organometallic reagent include n-butyl lithium, S-butyl reacting compound (2b-2) with chlorotriphenylmethane in lithium, t-butyl lithium and the like. Examples of the boronic the presence of a base. There are no particular limitations on acid ester include 2-methoxy-4,4,5,5-tetramethyl-1,3,2-di the solvent used in this reaction as long as it dissolves the 60 oxaborolane, trimethyl borate, triisopropyl borate and the starting materials to a certain extent without impeding the like. (1-Triphenylmethyl)-pyrazol-4-ylboronic acid, which is reaction. Examples of the solvent include ether solvents such produced in the case of using trimethylborate or triisopropyl as tetrahydrofuran and diethyl ether; amide solvents such as borate as the boronic acid ester, can be converted into a N,N-dimethylformamide and N-methylpyrrolidinone; and boronic acid pinacol ester in accordance with the literature dimethylsulfoxide, mixed solvents of the foregoing and the 65 (Journal of Heterocyclic Chemistry, Vol. 41, No. 6, 931 to like. Examples of the base include triethylamine, sodium 939, so as to obtain compound (2b-4). The organometallic hydride, potassium t-butoxide, potassium carbonate, sodium reagent is used in the amount of 1 to 1.5 equivalents based on US 8,158,657 B2 75 76 compound (2b-3). The boronic acid ester is used in the Manufacturing Method 2-2-2 Method 2 for Manufacturing amount of 1 to 1.5 equivalents based on compound (2b-3). Compound (2b) The reaction temperature for the an ionization reaction is from -90° C. to -60° C., and the reaction time is from 10 minutes to 24 hours. The temperature for the reaction with the boronic acid ester is from -78°C. to 0°C., with a reaction time being from 10 minutes to 12 hours. rs2 step2-9 Note that (1-triphenylmethyl)-pyrazol-4-yl boronic acid, R1 N R2 which is produced in the case of using trimethyl borate or (2b-6) triisopropylborate as the boronic acid ester in this reaction, 10 can be used in place of compound (2b-4) as the Substrate in Step 2-7. Step 2-7 This step is a step wherein compound (2b-5) is obtained by reacting compound (2b-4) with compound (1b-1) in the pres 15 ence of a palladium catalyst and a base. A phosphine ligand YO can be added to obtain good results. A quaternary ammonium M salt such as tetrabutylammonium bromide, tetrabutylammo ONB u nium chloride and the like can also be added in the amount of O 0.1 to 2 equivalents based on compound (2b-4). There are no particular limitations on the solvent used in this reaction as long as it dissolves the starting materials to a certain extent Hall I without impeding the reaction. Examples of the Solvent N (2b-4) 2 step2-10 include ether solvents such as 1,4-dioxane and tetrahydrofu 25 ran; aromatic hydrocarbon solvents such as benzene and tolu RI N R2 ene; amide solvents such as N,N-dimethylformamide and (2b-7) N-methylpyrrolidinone; alcohol solvents such as methanol and ethanol; and dimethylsulfoxide, water, mixed solvents of the foregoing and the like. Examples of the palladium catalyst 30 include palladium (II) acetate, tris(dibenzylidenacetone)di palladium (O), dichlorobis(triphenylphosphine)palladium (II), bis(tri-t-butylphosphine)palladium (O), tetrakis(triph enylphosphine)palladium (O), 1,1'-bis(diphenylphosphinof N O step2-11 errocene)dichloropalladium(II) and the like. Examples of the 35 N base include Sodium carbonate, potassium carbonate, cesium Hal N / carbonate, cesium fluoride, potassium phosphate, sodium hydroxide, potassium hydroxide and the like. Examples of 2 the phosphine ligand include triphenylphosphine, tri-t-bu RI N R2 tylphosphine, tricyclohexylphosphine, diphenylphosphinof 40 (2b-8) errocene, 2-dicyclohexylphosphinobiphenyl and the like. H The palladium catalyst is used in the amount of 0.01 to 0.3 N equivalents based on compound (2b-4). The base is used in N the amount of 1.5 to 10 equivalents based on compound Hal N M (2b-4). Compound (1b-1) is used in the amount of 1.0 to 3.0 45 equivalents based on compound (2b-4). The phosphine ligand 2 is used in the amount of 0.01 to 1.2 equivalents based on R1 N R2 compound (2b-4). The reaction temperature is from room (2b-9) temperature to reflux temperature, and the reaction time is from 10 minutes to 24 hours. 50 Step 2-8 (wherein R, R and Hal are defined as above.) This step is a step wherein compound (2b-1) is obtained by Compound (2b-6) which is a commercially available prod deprotecting the triphenylmethyl group of compound (2b-5) uct can be used as is, or may be manufactured from commer under acidic conditions. There are no particular limitations on cially available products by the known methods. Compound the solvent used in this reaction as long as it dissolves the 55 (2b-4) can be manufactured according to the methods starting materials to a certain extent without impeding the described in Manufacturing Method 2-2-1. reaction. Example of the solvent include ether solvents such Step 2-9 as 1,4-dioxane and tetrahydrofuran; aromatic hydrocarbon This step is a step wherein compound (2b-7) is obtained by Solvents such as benzene and toluene, alcohol solvents such Substituting a halogen atom for a hydrogenatom on the pyri as methanol and ethanol; methylene chloride, water, mixed 60 dine ring of compound (2b-6). Compound (2b-7) can be Solvents of the foregoing and the like. Examples of the acid manufactured according to the methods similar to those of include hydrochloric acid, Sulfuric acid, hydrobromic acid, Step 1-11. trifluoroacetic acid, formic acid and the like. The acid is used Step 2-10 in the amounts of from 2 equivalents to the solvent amount This step is a step wherein compound (2b-8) is obtained by based on compound (2b-5). The reaction temperature is from 65 reacting compound (2b-7) with compound (2b-4), in the pres 0° C. to reflux temperature, and the reaction time is from 10 ence of a palladium catalyst and a base. Compound (2b-8) can minutes to 24 hours. be manufactured according to the methods similar to those of US 8,158,657 B2 77 78 Step 2-7. With the proviso that compound (2b-4) is used in Step 2-13 the amount of 1 to 1.2 equivalents based on compound (2b-7). This step is a step wherein compound (2d-1) is obtained by Step 2-11 reacting compound (2c) with compound (2b-2). Compound (2d-1) can be manufactured according to the methods similar This step is a step wherein compound (2b-9) is obtained by to those of Step 2. deprotecting the triphenyl methyl group of compound (2b-8) Step 2-14 under acidic conditions. Compound (2b-9) can be manufac This step is a step wherein compound (2d) is obtained by tured according to the methods similar to those of Step 2-8. reacting compound (2d-1) with hexa(n-butyl)ditin in the Manufacturing Method 2-3 Method for Manufacturing presence of a palladium catalyst. A phosphine ligand can be Compound (2c): added into this reaction in order to obtain good results. There 10 are no particular limitations on the solvent used in this reac tion as long as it dissolves the starting materials to a certain extent without impeding the reaction. Examples of the Sol step2-12 vent include ether solvents such as 1,4-dioxane and tetrahy drofuran; aromatic hydrocarbon Solvents such as toluene and 15 xylene; amide solvents such as N,N-dimethylformamide and Z N-methylpyrrolidinone; dimethyl sulfoxide, mixed solvents of the foregoing and the like. Examples of the palladium 1O- NR3 catalyst include palladium (II) acetate, tris(dibenzylidenac etone)dipalladium (O), dichlorobis(triphenylphosphine)pal (2c) ladium (II), dichlorobis(tri-o-tolylphosphine)palladium (II), tetrakis(triphenylphosphine)palladium (O) and the like. (wherein ring A. L. R. and Z are defined as above.) Examples of the phosphine ligand include triphenylphos Compound (1c-3) which is a commercially available prod phine, tri-o-tolylphosphine and the like. The hexa(n-butyl) uct can be used as is, or compound (1c-3) can also be manu ditin is used in the amount of 1 to 10 equivalents, preferably factured from commercially available products by the well 25 3 to 5 equivalents, based on compound (2d-1). The palladium catalyst is used in the amount of 0.01 to 0.3 equivalents based known methods. Compound (1c-3) can further be manufac on compound (2d-1). The phosphine ligand is used in the tured according to the methods described in the Manufactur amount of 0.01 to 1.2 equivalents based on compound (2d-1). ing Examples in the Examples, Manufacturing Method 1-3- The reaction temperature is from room temperature to reflux 1 given above or the like. temperature, and the reaction time is from 10 minutes to 48 Step 2-12 30 hours. This step is a step wherein compound (2c) is obtained by Compound (2d) can also be obtained from compound (2d converting the hydroxyl group of compound (1c-3) into a 1) according to the method given below as Alternative leaving group. Compound (2c) can be manufactured accord Method (1). ing to the methods similar to those of Step 1-32. Alternative Method (1): Compound (2d) can be obtained by 35 first an ionizing the bromine atom of compound (2d-1) using Manufacturing Method 2-4 Method for Manufacturing an organometallic reagent, and then reacting with tri(n-butyl) Compound (2d): tin chloride. There are no particular limitations on the solvent used in this reaction as long as it dissolves the starting mate rials to a certain extent without impeding the reaction. NH 40 Examples of the solvent include ether solvents such as tet Br rahydrofuran and diethyl ether; aromatic hydrocarbon sol eN vents such as benzene and toluene; hexane, mixed solvents of Z (2b-2) the foregoing and the like. Examples of the organometallic I1O- NR3 step2-13 reagent include n-butyl lithium, s-butyl lithium, t-butyl 45 lithium and the like. The organometallic reagent is used in the (2c) amount of 1 to 1.5 equivalents based on compound (2d-1). Z The tri(n-butyl)tin chloride is used in the amount of 1 to 1.5 equivalents based on compound (2d-1). The reaction tem Br /S1G)-- R step2-14 - perature for the an ionization reaction is from -90° C. to -60° 50 C., with a reaction time being from 10 minutes to 24 hours. (2d-1) The temperature for the reaction with the tri(n-butyl)tin chlo n-Bu n-Bu ZN ride is from -78°C. to 0°C., with a reaction time being from \ 7 N rO R3 10 minutes to 12 hours. / leN Manufacturing Method 3 Typical Method for Manufactur n-Bu 55 ing Compound (3a): (2d) Z (3a) YR3 (wherein ring A. L. R. and Z are defined as above.) Compound (2c) which is a commercially available product 60 e can be used as is, or compound (2c) can also be manufactured from commercially available products by the well known N N NN/ methods. Compound (2c) can further be manufactured according to the methods described in the Manufacturing Examples in the Examples, Manufacturing Method 2-3 65 R1 N R2 given above or the like. A commercial product may be used as is for compound (2b-2). (wherein ring A, R', R, R and Z are defined as above.) US 8,158,657 B2 79 80 Manufacturing Method 3-1 Method for Manufacturing -continued Compound (3a): Br

e Br 5 M e A r’s2 N NN N R1 N R2 10 (3.b) 2 RI N R2 (3.b) (wherein RandR are defined as above, and R“represents a n-Bu hydrogen atom and NHR'. R” represents a protective Sn N3 - - 15 group Such as t-butoxycarbonyl, t-butylcarbonyl and the n-B^N (NOY-B R step3tep3 like.) Compound (3b-1) which is a commercially available prod (3.c) uct can be used as is, or compound (3b-1) can also be manu Z YR3 factured from commercially available products by the well 2O known methods. A commercial product can be used as is for e compound (2b-1). M Step 3-1 N NN N This step is a step wherein compound (3b-2) is obtained by reacting compound (3b-1) with compound (2b-1) in the pres 2 25 ence of a base and a copper catalyst. A copper ligand can also RI N R2 be added to improve the yield. There are no particular limi (3a) tations on the solvent used in this reaction as long as it dis Solves the starting materials to a certain extent without imped ing the reaction. Examples of the solvent include ether (wherein ring A. R', R, R and Z are defined as above.) 30 Solvents such as 1,4-dioxane and tetrahydrofuran, aromatic Compound (3.b) can be manufactured from commercially hydrocarbon solvents such as benzene, toluene and Xylene; available products by the well known methods, or compound amide solvents such as N,N-dimethylformamide and N-me (3.b) can also be manufactured according to the methods given thylpyrrolidinone; and dimethylsulfoxide, mixed solvents of in the Manufacturing Examples in the Examples, Manufac 35 the foregoing and the like. Examples of the base used in this turing Method 3-2 given below or the like. reaction include potassium carbonate, cesium carbonate, Compound (3.c) can be manufactured from a commercially potassium phosphate, potassium t-butoxide, Sodium t-butox available product by the well known methods, or compound ide and the like. Examples of the copper catalyst include (3.c) can also be manufactured according to the methods given copper(I) iodide, copper(I) bromide, copper(I) chloride and in the Manufacturing Examples in the Examples, Manufac 40 the like. Examples of the copperligand include 1.2-cyclohex turing Method 3-3 given below or the like. anediamine, N,N-dimethyl-cyclohexane-1,2-diamine, 1.10 Step 3 phenanthroline and the like. Compound (2b-1) is used in the This step is a step wherein compound (3a) is obtained by amounts of from 1 to 5 equivalents based on compound (3b reacting compound (3.b) with compound (3c). Compound 1). The base is used in the amount of 1 to 5 equivalents based (3.a) can be manufactured according to the methods similar to 45 on compound (3b-1). The copper catalyst is used in the those of Step 1-8. amount of 0.01 to 0.3 equivalents based on compound (3b-1). Manufacturing Method 3-2 Method for Manufacturing The copper ligand is used in the amount of 1 to 3 equivalents Compound (3b): based on the copper catalyst. The reaction temperature is from 50° C. to reflux temperature, and the reaction time is 50 from 30 minutes to 48 hours. I Br Step 3-2 N This step is a step wherein compound (3.b) is obtained by a step3-1step3 reacting acid with compound (3b-2). So as to deprotect an 2 amine moiety. There are no particular limitations on the Sol RI N R2a HNN/ 55 vent used in this reaction as long as it dissolves the starting (3b-1) (2b-1) materials to a certain extent without impeding the reaction. Br Examples of the solvent include alcohol solvents such as methanol and ethanol; and water, mixed solvents of the fore e 60 going and the like. Examples of the acid include inorganic acids Such as hydrochloric acid, Sulfuric acid, and hydrobro N NN / step3-2 mic acid; organic acids such as trifluoroacetic acid, p-tolu enesulfonic acid; and the like. The acid is used in the amounts RI 4. R2a of from 2 equivalents to the solvent amount based on com 65 pound (3b-2). The reaction temperature is from room tem (3b-2) perature to reflux temperature, and the reaction time is from 30 minutes to 72 hours. US 8,158,657 B2 81 82 Manufacturing Method 3-3 Method for Manufacturing Manufacturing Method 4-1 Method for Manufacturing Compound (3.c): Compound (4a):

n R3

e O He 10 C N N/ step4

2 N NH2 (4a-1) 15 Z n R3

e

S. /9 N N

2 N NH2 (wherein ring A. L. R and Z are defined as above.) (4a) Compound (2c) can be manufactured from commercially 25 available products by the well known methods, or compound (2c) can also be manufactured according to the methods (wherein ring A, Rand Z are defined as above.) described in the Manufacturing Examples in the Examples, Compound (4a-1) can be manufactured from commer Manufacturing Method 2-3 given above or the like. cially available products by the well known method, or com Step 3-3 30 pound (4a-1) can be manufactured according to the methods This step is a step wherein compound (3.c) is obtained by described in the Manufacturing Examples in the Examples, reacting compound (2c) with tributyltin-anions. There are no Manufacturing Method 4-2 given below or the like. particular limitations on the solvent used in this reaction as Step 4 long as it dissolves the starting materials to a certain extent 35 This step is a step wherein compound (4a) is obtained by without impeding the reaction. Examples of the Solvent Substituting a hydrogen atom for a chlorine atom of com include ether solvents such as tetrahydrofuran and diethyl pound (4a-1). Compound (4a) can be obtained by reacting ester, aromatic hydrocarbon solvents such as benzene, tolu compound (4a-1) in the presence of a palladium catalyst, a ene and Xylene; mixed solvents of the foregoing and the like. base and a hydrogen Source. A phosphine ligand can also be The tributyltin-anions used in the reaction can be synthesized 40 added to obtain good results. There are no particular limita by reacting an organometallic reagent with tributyltin tions on the solvent used in this reaction as long as it dissolves hydride. Examples of the organometallic reagent include the starting materials to a certain extent without impeding the lithium diisopropylamide, isopropyl magnesium chloride, reaction. Examples of the solvent include ether solvents such methyl magnesium iodide and the like. The tributyltin 45 as 1,4-dioxane and tetrahydrofuran; aromatic hydrocarbon hydride is used in the amount of 1 to 2 equivalents based on Solvents such as toluene and Xylene; amide solvents such as compound (2c). The organometallic reagent is used in the N,N-dimethylformamide and N-methylpyrrolidinone; and amount of 1 to 1.5 equivalents based on tributyltin hydride. dimethyl sulfoxide, mixed solvents of the foregoing and the The reaction temperature is from -78°C. to room tempera like. Examples of the palladium catalyst include bis(tri-t- ture, and the reaction time is from 10 minutes to 12 hours. 50 butylphosphine)palladium(0), palladium(II) acetate, tetrakis Manufacturing Method 4 Typical Method for Manufactur (triphenylphosphine)palladium (O), dichlorobis(triph ing Compound (4a): enylphosphine)palladium (II), tris(dibenzylidenacetone) dipalladium (O) and the like. Examples of the base include triethylamine, N,N-diisopropylethylamine and the like. (4a) 55 Examples of the hydrogen Source include formic acid, potas sium formate, sodium formate, lithium formate, ammonium ^ns formate and the like. Examples of the phosphine ligand e include triphenylphosphine, tri-o-tolylphosphine, tri-t-bu tylphosphine and the like. The palladium catalyst is used in N s o 60 the amount of 0.01 to 0.3 equivalents based on compound (4a-1). The base is used in the amount of 2 to 5 equivalents 2 based on compound (4a-1). The hydrogen source is used in N R2 the amount of 1 to 5 equivalents based on compound (4a-1). The phosphine ligand is used in the amount of 0.01 to 1.2 65 equivalents based on compound (4a-1). The reaction tem perature is from room temperature to reflux temperature, and (wherein ring A. R. Rand Z are defined as above.) the reaction time is from 30 minutes to 24 hours. US 8,158,657 B2 83 84 Manufacturing Method 4-2 Method for Manufacturing Step 4-2 Compound (4a-1): This step is a step wherein compound (4b-3) is obtained by reacting compound (4b-2) with a chlorinating agent. There are no particular limitations on the solvent used in this reac O tion as long as it dissolves the starting materials to a certain extent without impeding the reaction. Examples of the Sol N He vent include ether solvents such as 1,4-dioxane and tetrahy step4-1 drofuran; alcohol solvents such as methanol and ethanol, 2 amide solvents such as N,N-dimethylformamide and N-me N NH2 thylpyrrolidinone; and dimethyl sulfoxide, methylene chlo 10 rine, water, mixed solvents of the foregoing and the like. (4b-1) Examples of the chlorinating agent include N-chlorosuccin OH imide, sodium hypochlorite, chlorine and the like. The chlo rinating agent is used in the amount of 2 to 5 equivalents based on compound (4b-2). The reaction temperature is from 0°C. N H step4-2 15 to room temperature, and the reaction time is from 10 minutes to 24 hours. 2 Step 4-3 N NH2 This step is a step wherein compound (4a-1) is obtained by (4b-2) reacting compound (4b-3) with compound (4c). Compound OH ^n, (4a-1) can be manufactured according to the methods similar R to those of Step 1. Manufacturing Method 4-3 Method for Manufacturing C --Or(4c) Compound (4c): N C step4-3 25 2 Z N NH2 n (4b-3) 11-O- R3 step4-4 Z n R3 (2c) 30 Z n er R e1O- R3 /9 Pi step4-5 C N s R6 2 (4c-1) N NH2 Z (4a-1) e1G)1 NR3

40 (4c) (wherein ring A. R. and Z are as defined above.) Compound (4b-1) which is a commercially available prod (wherein ring A. R. R. R. L. and Z are defined as above.) uct can be used as is. Compound (4c) can be manufactured Compound (2c) can be manufactured from commercially from commercial available products by the well known meth available products by the well known methods, or compound ods, or compound (4c) can also be manufactured according to 45 (2c) can also be manufactured according to the methods the methods described in the Manufacturing Examples in the described in the Manufacturing Examples in the Examples, Examples, Manufacturing Method 4-3 given below or the Manufacturing Method 2-3 given below or the like. like. Step 4-4 This step is a step wherein compound (4c-1) is obtained by Step 4-1 50 reacting compound (2c) with an ethynylsilane derivative. This step is a step wherein compound (4b-2) is obtained by Compound (4c-1) can be obtained by reacting compound (2c) reacting compound (4b-1) with hydroxylamine or hydroxy with an ethynyl Grignard reagent obtained by reacting an lamine hydrochloride in the presence of a base. There are no ethynyl silane derivative with a Grignard reagent. A copper particular limitations on the solvent used in this reaction as reagent such as copper(I) bromide, copper(I) iodide and the long as it dissolves the starting materials to a certain extent 55 like can also be added to obtain good results. Examples of the without impeding the reaction. Examples of the Solvent ethynylsilane derivative include trimethylsilyl acetylene, tri include alcohol solvents such as methanol and ethanol; and ethylsilyl acetylene, triisopropylsilyl acetylene, t-butyldim methylene chloride, water and the like. The base can also be ethylsilyl acetylene and the like. An alkyl magnesium halide used as the solvent. Examples of the base include pyridine, Such as ethyl magnesium bromide and isopropyl magnesium Sodium hydroxide, potassium hydroxide, Sodium acetate, 60 chloride can be used as the Grignard reagent. The ethynylsi Sodium carbonate, Sodium hydrogen carbonate and the like. lane derivative can be used in the amount of 1 to 3 equivalents Hydroxylamine or hydroxylamine hydrochloride is used in based on compound (2c). The Grignard reagent can be used in the amounts of from 1 to 10 equivalents based on compound the amount of 1 to 3 equivalents based on compound (2c). The (4b-1). The base is used in the amount of 1 equivalent to the copper reagent can be used in the amount of 0.1 to 3 equiva Solvent amount based on compound (4b-1). The reaction 65 lents based on compound (2c). The reaction temperature is temperature is from 0° C. to reflux temperature, and the from room temperature to reflux temperature, and the reac reaction time is from 10 minutes to 24 hours. tion time is from 1 hour to 72 hours. US 8,158,657 B2 85 86 Step 4-5 compound (5a-1). The reaction temperature is from room This step is a step wherein compound (4c) is obtained by temperature to reflux temperature, and the reaction time is deprotecting the trimethylsilyl group of compound (4c-1). from 10 minutes to 48 hours. Compound (4c) can be manufactured according to the meth Compound (5a) can also be obtained from compound (5a ods similar to those of Step 1-2. 5 1) according to the method described below as Alternative Manufacturing Method 5 Typical method for manufactur Method (1). ing compound (5a): Alternative Method (1): Compound (5a) can be obtained by reacting compound (5a-1) with compound (1c-10-2) in the presence of a base. There are no particular limitations on the (5a) 10 Solvent used in this reaction as long as it dissolves the starting materials to a certain extent without impeding the reaction. 21 Examples of the solvent include ether solvents such as tet 4 cyA -o rahydrofuran and diethyl ether, aromatic hydrocarbon sol R ? N \-R vents such as benzene and toluene, amide solvents such as 15 N,N-dimethylformamide and N-methylpyrrolidinone; and 2 dimethyl sulfoxide, mixed solvents of the foregoing and the RI N R2 like. Examples of the base include Sodium hydride, potassium carbonate, Sodium carbonate, cesium carbonate, potassium (wherein R. R. R. R. X and Y are defined as above.) hydroxide, Sodium hydroxide and the like. A catalytic amount Manufacturing Method 5-1 Method for Manufacturing of sodium iodide or potassium iodide or tetrabutylammonium Compound (5a): iodide can also be added to obtain good results. The base is LN-R (1c-10-2) X X 21 n 21 -H OH / step5-1) Y. 4 Ky\ Jo R N KW li N HO R3 R N1 N \-R O2 N (1c-10-1)N1 / 2 RI N R2 step5-2 R1 N R2 (5a-1) (5a)

35 (wherein R. R. R. R. X and Y are defined as above.) added in the amount of 1 to 1.5 equivalents based on com Compound (5a-1) can be manufactured according to the pound (5a-1). The reaction temperature is from room tem method described in the Manufacturing Examples in the perature to reflux temperature, and the reaction time is from Examples, Manufacturing Method 5-2 given below or the 10 minutes to 48 hours. like. Compound (1c-10-1) and compound (1c-10-2) which 40 Step 5-2 are commercially available products can be used as is, or they This step is a step wherein compound (5a) is obtained by can also be manufactured from commercially available prod reacting compound (5a-1) with compound (1c-10-1). Com ucts by the well known methods. pound (5a) can be manufactured according to the methods Step 5-1 similar to those of Step 1-37. This step is a step wherein compound (5a) is obtained by 45 Manufacturing Method 5-2 Method for Manufacturing adding 1 equivalent of base to compound (5a-1) to obtain Compound (5a-1): phenoxide ions, followed by reacting with compound (1c-10 2). Phenoxide ion production: Phenoxide ions can be obtained X 4N by adding 1 equivalent of a base to compound (5a-1) in a 50 solvent such as tetrahydrofuran or methanol. Examples of the base include potassium hydroxide, Sodium hydroxide, potas sium carbonate, sodium carbonate, potassium t-butoxide and 2 the like, preferably sodium hydroxide. The solvent is prefer R1 N R2 ably concentrated for use in the following reaction. The reac 55 (5a-2) tion temperature is room temperature, and the reaction time is from 5 minutes to 1 hour. XN 21 Reaction of phenoxide ions with compound (1c-10-2): The 4 K i --OH phenoxide ions and compound (1c-10-2) are reacted in a R N1 I N Solvent to obtain compound (5a). There are no particular 60 2 limitations on the solvent used in this reaction as long as it RI N R2 dissolves the starting materials to a certain extent without impeding the reaction. Example of the solvent include amide (5a-1) solvents such as N,N-dimethylformamide, N-methylpyrroli dinone and hexamethylphosphoramide; and dimethylsulfox 65 (wherein R. R. R. X and Y are defined as above.) ide, mixed solvents of the foregoing and the like. Compound Compound (5a-2) can be manufactured according to the (1c-10-2) is used in the amount of 1 to 3 equivalents based on methods described in the Manufacturing Examples in the US 8,158,657 B2 87 88 Examples, Manufacturing Method 1). Manufacturing Step 6-1 Method 2. Manufacturing Method 3 and Manufacturing This step is a step wherein compound (6a-1) is obtained by Method 4 which are given above or the like. Substituting a halogen atom for a hydrogenatom on the pyri Step 5-3 dine ring of compound (6a-2). Compound (6a-1) can be 5 manufactured according to the methods similar to those of This step is a step wherein compound (5a-1) is obtained by Step 1-11. reacting acid with compound (5b-2). An additive Such as Manufacturing Method 6-2 Method 2 for Manufacturing thioanisole may be added in the reaction system to obtain Halogen-Modified Product of Compound (1a) better results. There are no particular limitations on the sol vent used in this reaction as long as it dissolves the starting materials to a certain extent without impeding the reaction. Examples of the solvent include ether solvents such as diethyl ether and tetrahydrofuran; and methylene chloride, trifluoro acetic acid and the like. Examples of the acid include organic acids Such as trifluoroacetic acid and methanesulfonic acid; inorganic acids Such as Sulfuric acid; Lewis acids such as boron trifluoride diethyl etherate; and the like. Examples of the additives include thioanisole, ethanethiol, d1-methionine and the like. The acid is used in the amount of 1 equivalent to the solvent amount based on compound (5a-2). The additive is used in the amount of 1 to 5 equivalents based on compound (5a-2). The reaction temperature is from 0°C. to reflux tem perature, and the reaction time is from 10 minutes to 72 hours. Compound (5a-1) can also be obtained from compound (5a-2) according to the method described below as Alterna 25 tive Method (1). Alternative Method (1): Compound (5a-1) can be obtained by reacting compound (5a-2) with boron tribromide or boron trichloride. There are no particular limitations on the solvent used in this reaction as long as it dissolves the starting mate 30 (6a-3) rials to a certain extent without impeding the reaction, but methylene chloride is preferably used. The borontribromide (wherein ring A. Z. Hal, R and Rare defined as above; R' or borontrichloride is used in the amount of 1 to 5 equivalents represents a hydrogen atom or a C- alkyl group.) based on compound (5a-2). The reaction temperature is from 35 Compound (6a-4) can be manufactured according to the -78°C. to room temperature, and the reaction time is from 30 methods described in Manufacturing Method 1. minutes to 24 hours. Step 6-2 Manufacturing Method 6-1 Method 1 for Manufacturing This step is a step wherein compound (6a-3) is obtained by Halogen-Modified Product of Compound (1a) Substituting a halogen atom for a hydrogenatom on the pyri dine ring of compound (6a-4). Compound (6a-3) can be 40 manufactured according to the methods similar to those of Step 1-11. Manufacturing Method 7 Method 3 for Manufacturing Halogen-Modified Product of Compound (1a) 45

50 (6a-2)

55 Hal

60 (6a-1) (wherein ring A. Z. Hal, R' and Rare defined as above; R. represents a hydrogen atom or a C- alkyl group.) 65 Compound (6a-2) can be manufactured according to the methods described in Manufacturing Method 1. US 8,158,657 B2 89 90 -continued Manufacturing Method 8) Method 1 for Manufacturing Amino Group-Modified Product of Compound (1a)

Z N R3

C \ O N 2 H6 R-( F 2 10 N (1c-1) R (S f 3 H step7-4) N1,N-1 - (8a-1-1) 2 step8 N NH2 2 HN N R2 (7a-4) 15 (8a-1)

R (S f R3 25 N1,N1i 71

351NN N2 R2 H (8a) 30

(wherein ring A. R. R. R. X, Y and Z are defined as above; R represents a hydrogenatom, a Cls alkyl group, a hydroxy 35 Cs alkyl group, a C alkoxy carbonyl group or a C (wherein ring A, Hal, R. R. R. and Z is defined as above.) alkoxy Cs alkyl group.) Compound (8a-1-1) which is a commercially available Compound (7a-1) which is a commercially available can product can be used as is, or may also be manufactured from be used as is. Compound (1c-1) can be manufactured from commercially available products by the known methods. commercially available products by the known methods, and Compound (8a-1) can be manufactured according to the can be manufactured according to the methods described in 40 methods described in Manufacturing Method 1), or the like. Manufacturing Example of Example or Manufacturing Step 8 Method 1-3-1, and the like. This step is a step wherein compound (8a) is obtained by reacting compound (8a-1) and compound (8a-1-1) in the Step 7-1 presence of a reducing agent. This step can be carried out by This step is a step wherein compound (7a-2) is obtained by 45 adding an acid such as acetic acid or hydrochloric acid in Substituting a halogen atom for a hydrogenatom on the pyri catalytic amount to solvent amount. There are no particular dine ring of compound (7a-1). Compound (7a-2) can be limitations on the solvent used in this reaction as long as it manufactured according to the methods similar to those of dissolves the starting materials to a certain extent without Step 1-11. impeding the reaction. Examples of the Solvent include ether 50 Solvents such as tetrahydrofuran and diethyl ether, aromatic Step 7-2 hydrocarbon solvents such as benzene and toluene; amide This step is a step wherein compound (7a-3) is obtained by solvents such as N,N-dimethylformamide and N-methylpyr reacting compound (7a-2) and an ethinylsilane derivative. rolidinone; alcohol Solvents such as methanol and ethanol: Compound (7a-3) can be manufactured according to the halogenated hydrocarbon solvents such as methylene chlo methods similar to those of Step 1-1. 55 ride and chloroform, 1,2-dichloroethane; water, acetic acid, mixed solvents of the foregoing, or the like, preferably a Step 7-3 mixed solvent of N,N-dimethylformamide and acetic acid. This step is a step wherein compound (7a-4) is obtained by Examples of the reducing agent used in this reaction include reacting compound (7a-3) with a base. Compound (7a-4) can a-picoline borane, pyridine-borane, Sodium cyanoborohy be manufactured according to the methods similar to those of 60 dride, sodium triacetoxyborohydride and the like, preferably Step 1-2. C-picoline borane. Compound (8a-1-1) can be used in the amount of 1 to 5 equivalents based on compound (8a-1), Step 7-4 preferably 1 to 1.5 equivalents. The reducing agent can be This step is a step wherein compound (7a) is obtained by used in the amount of 0.5 to 5 equivalents based on compound reacting compound (7a-4) and compound (1c-1), in the pres 65 (8a-1), preferably 1 to 1.5 equivalents. The reaction tempera ence of a base. Compound (7a) can be manufactured accord ture is from 0°C. to reflux temperature, and the reaction time ing to the methods similar to those of Step 1. is from 10 minutes to 48 hours. US 8,158,657 B2 91 92 Manufacturing Method 9 Method 2 for Manufacturing Amino Group-Modified Product of Compound (1a)

O O O .. O ---. (8a-1-2) (8a-1-3) (Sy step9-1 R N1,N-1 71 R3 lsO R36 OH 2 2 HN N R (8a-1-4) ---step9-2 (8a-1)

R (6 S.f R3 O N1,N1 71

R36 ls N N 2 R2

(9a)

30 (wherein ring A. R. R. R. X,Y and Zare defined as above; impeding the reaction. Examples of the solvent include halo R represents a C-alkyl group or a Calkoxyalkyl group.) genated hydrocarbon solvents such as methylene chloride Compound (8a-1-2), compound (8a-1-3) and compound and chloroform; ether solvents such as tetrahydrofuran and (8a-1-4) which are commercially available products can be 1,4-dioxane: amide solvents such as N,N-dimethylforma used as is, or they may also be manufactured from commer 35 mide and N-methylpyrrolidinone; sulfoxide solvents such as cially available products by the known methods. Compound dimethyl sulfoxide; ester solvents such as ethyl acetate, (8a-1) can be manufactured according to the methods mixed solvents of the foregoing, or the like. Examples of the described in Manufacturing Method 1), or the like. condensing reagent include Bop (1H-1,2,3-benzotriazole-1- Step 9-1 yloxy(tri(dimethylamino))phosphonium hexafluorophos This step is a step wherein compound (9a) is obtained by 40 phate), WSC (1-ethyl-3-(3-dimethylaminopropyl)carbodi reacting compound (8a-1-2) or compound (8a-1-3) with com imide hydrochloride), DCC (N,N- pound (8a-1), in the presence of a base. There are no particu dicyclohexylcarbodiimide), or the like. A catalytic amount of lar limitations on the solvent used in this reaction as long as it 4-dimethylaminopyridine can also be added to accelerate the dissolves the starting materials to a certain extent without reaction. In addition, this step can also be carried out by impeding the reaction. Examples of the Solvent include ether 45 adding a base such as triethylamine in the amount of 1 to 5 Solvents such as tetrahydrofuran and diethyl ether, aromatic equivalents. Compound (8a-1-4) can be used in the amount of hydrocarbon solvents such as benzene and toluene; amide 1 to 3 equivalents based on compound (8a-1), preferably 1 to solvents such as N,N-dimethylformamide and N-methylpyr 1.5 equivalents. The condensing reagent can be used in the rolidinone; halogenated hydrocarbon Solvents such as meth amount of 1 to 3 equivalents based on compound (8a-1), ylene chloride and chloroform, mixed solvents of the forego 50 preferably 1 to 1.5 equivalents. The reaction temperature is ing, or the like. Examples of the base include triethylamine, from 0°C. to reflux temperature, and the reaction time is from pyridine, potassium carbonate, or the like. A catalytic amount 10 minutes to 48 hours. of 4-dimethylaminopyridine can also be added in order to Manufacturing Method 10 Method 3 for Manufacturing accelerate the reaction. Compound (8a-1-2) or compound Amino Group-Modified Product of Compound (1a) (8a-1-3) can be used in the amount of 1 to 5 equivalents based 55 on compound (8a-1), preferably 1 to 1.5 equivalents. The base can be used in the amount of 0.5 equivalents to solvent amount based on compound (8a-1), preferably 1 to 1.5 O equivalents. The reaction temperature is from 0°C. to reflux temperature, and the reaction time is from 10 minutes to 48 & H hours. R (6 Y R3 Step 9-2 f 71 (10a-1-1) This step is a step wherein compound (9a) is obtained by step 10 2 reacting compound (8a-1) and compound (8a-1-4), in the R37 N NH2 presence of a condensing reagent. There are no particular 65 limitations on the solvent used in this reaction as long as it dissolves the starting materials to a certain extent without US 8,158,657 B2 93 94 -continued saturated aqueous Sodium chloride, and the Solvent was X N. evaporated under a reduced pressure. The residue was puri R (6. Y R3 fied by NH silica gel column chromatography (ethyl acetate: N1,N1 71 heptane=1:3) to obtain the title compound (240 mg, 15%). 'H-NMR Spectrum (CDC1) & (ppm): 4.00 (2H, s), 5.05 R37 21N1 as (2H, s), 5.41 (2H, s), 6.24 (1H, s), 6.71 (1H, dd, J=4.9, 7.6 Hz), 6.93-6.97 (2H, m), 7.18-7.22 (2H, m), 7.31-744 (5H, m), 7.70 (1H, dd, J=1.7, 7.6Hz), 8.13 (1H, dd, J=1.8, 4.9 Hz). s The starting material, 4-benzyloxy-phenyl-acetohydroxi 10 (10a) moyl chloride, was synthesized as follows. (whereinring A. R. R. X, Y and Zare defined as above; R. Manufacturing Example 1-1-1 represents a hydrogen atom, a halogen atom, R' (CO)— 1-Benzyloxy-4-((E)-2-nitro-vinyl)-benzene NH-(R' is a C- alkyl group or a C- alkoxy C, alkyl 15 group), a C- alkyl group, a hydroxy Co alkyl group, a cyano Ce alkyl group, a Calkoxy group or a C- alkoxy C- alkyl group; R represents a hydrogen atom and a Cls alkyl group.) Compound (10a-1-1) which is be a commercially available product can be used as is, or may also be manufactured from the commercially available products by the known methods. Compound (10a-1) can be manufactured according to the methods described in Manufacturing Method 1), or the like. Step 10 25 This step is a step wherein compound (10a) is obtained by reacting compound (10a-1) and compound (10a-1-1) in the To a mixture of 4-benzyloxybenzaldehyde (1.0 g, 4.7 presence of a reducing agent. Compound (10a) can be manu mmol) and sodium methoxide (28% methanol solution, 150 factured according to the methods similar to those of Step 8. uL, 0.74 mmol) and methanol (10 mL) were added 30 nitromethane (330 uL, 6.1 mmol) and sodium methoxide EXAMPLES (28% methanol solution, 1.0 mL, 4.9 mmol) at 0°C., which was stirred for 10 minutes at room temperature. The reaction The compounds according to the present invention can be mixture was cooled to 0°C., and 5 Naqueous hydrochloric manufactured, for example, according to the methods acid solution (20 mL) was added thereto at the same tempera described in the following manufacturing examples and 35 ture. The reaction mixture was then stirred for 15 minutes at examples. These are only examples, however, and the com room temperature. The precipitated solids were filtered to pounds according to the present invention are in no way obtain the title compound (1.2g, 100%). limited to the following specific examples. H-NMR Spectrum (DMSO-d) & (ppm): 5.20 (2H, s), Example 1 40 7.10-7.14 (2H, m), 7.32-7.48 (5H, m), 7.82-7.85 (2H, m), 8.12 (2H, dd, J=13.5, 18.2 Hz). 3-(3-(4-Benzyloxy-benzyl)-isoxazol-5-yl)-pyridin-2- ylamine Manufacturing Example 1-1-2

45 1-Benzyloxy-4-(2-nitro-ethyl)-benzene

50

55 To a mixture of 1-benzyloxy-4-((E)-2-nitro-vinyl)-ben Zene (1.0 g, 3.9 mmol) described in Manufacturing Example To a mixture of 4-benzyloxy-phenyl-acetohydroximoyl 1-1-1, acetic acid (1 mL) and dimethylsulfoxide (17 mL) was chloride (1.2 g, 4.4 mmol) described in Manufacturing 60 added sodium borohydride (250 mg. 6.3 mmol) at room tem Example 1-1-3 and tetrahydrofuran (34 mL) were added perature while cooling appropriately, and the reaction mix 3-Ethynyl-pyridin-2-ylamine (260 mg, 2.2 mmol) described ture was stirred for 40 minutes at room temperature. Water in Manufacturing Example 1-2-3 and triethylamine (3.0 mL. was added to the reaction mixture. The reaction mixture was 22 mmol) at 0°C., which was stirred for 1 hour at room partitioned into ethylacetate and water. The organic layer was temperature. To the reaction mixture was added water at room 65 washed with water and Saturated aqueous sodium chloride, temperature, which was then extracted with ethyl acetate and dried over anhydrous magnesium Sulfate, and the solvent tetrahydrofuran (2:1). The organic layer was washed with was evaporated under a reduced pressure. The residue was US 8,158,657 B2 95 96 purified by NH silica gel column chromatography (ethyl sodium sulfate. The sodium sulfate was removed by filtration, acetate:heptane=1:3) to obtain the title compound (710 mg. and the solvent was concentrated under a reduced pressure to 70%). obtain the title compound (41.2g, 85.9%). H-NMR Spectrum (CDC1) & (ppm): 3.26 (2H, t, J–7.2 H-NMR Spectrum (DMSO-d) & (ppm): 6.00 (2H, brs), Hz), 4.56 (2H, t, J–7.2 Hz), 5.04 (2H, s), 6.92 (2H, d, J–8.4 6.32 (1H, dd, J=4.8 Hz, 7.2 Hz), 7.87 (1H, d, J–7.2 Hz), 7.92 Hz), 7.11 (2H, d, J=8.8 Hz), 7.30-7.42 (5H, m). (1H, d, J–4.8 Hz). Manufacturing Example 1-1-3 Manufacturing Example 1-2-2 4-Benzyloxy-phenyl-acetohydroximoyl chloride 10 3-Trimethylsilanylethynyl-pyridin-2-ylamine

15

N NH2

To a mixture of 1-benzyloxy-4-(2-nitro-ethyl)-benzene To a mixture of 3-iodopyridin-2-ylamine (40.2 g, 183 (340 mg, 1.3 mmol) described in Manufacturing Example mmol) described in Manufacturing Example 1-2-1, trimeth 1-1-2 and methanol (5 mL) was added lithium methoxide 25 ylsilylacetylene (51.7 mL,366 mmol), copper(I) iodide (3.49 (100 mg, 2.6 mmol) at room temperature, which was stirred g, 18.3 mmol), N,N-diisopropylethylamine (63.7 mL, 366 for 15 minutes at room temperature. The reaction mixture was mmol) and N-methylpyrrolidinone (200 mL) was added tet concentrated under a reduced pressure. Methylene chloride rakis(triphenylphosphine)palladium (O) (10.6 g., 9.15 mmol) (4 mL) and tetrahydrofuran (2 mL) were added to the residue. under nitrogen atmosphere, which was stirred for 3 hours and Titanium (IV) chloride was added at 78° C. to the reaction 30 10 minutes at room temperature. Water was added to the mixture, which was then stirred for 50 minutes at 0°C. The reaction solution, which was then extracted with ethyl acetate reaction mixture was cooled to -78° C., and after adding 4 times. The solvent was concentrated under a reduced pres water (5 mL), the reaction mixture was gradually warmed to sure. The residue was purified by NH silica gel chromatog room temperature. The reaction mixture was partitioned into raphy (heptane:ethylacetate 4:1). The resulting solution was ethyl acetate and water. The organic layer was washed with 35 concentrated under a reduced pressure, and the residue was saturated aqueous Sodium chloride, and the Solvent was purified by silica gel chromatography (heptane:ethyl evaporated under a reduced pressure. The residue was puri acetate-2:1 then 1:1) to obtain the title compound (28.1 g, fied by neutral silica gel column chromatography (ethyl 80.7%). acetate:heptane=1:3) to obtain the title compound (310 mg. 40 H-NMR Spectrum (DMSO-d) & (ppm): 0.25 (9H, s), 84%). 6.09 (2H, brs), 6.51-6.57 (1H, m), 7.50-7.55 (1H, m), 7.95 H-NMR Spectrum (CDC1) & (ppm): 3.83 (2H, s), 5.07 7.99 (1H, m). (2H, s), 6.94-6.98 (2H, m), 7.17-7.21 (2H, m), 7.32-7.44 (5H, m). Manufacturing Example 1-2-3 The starting material, 3-ethynyl-pyridin-2-ylamine, was synthesized as follows. 45 3-Ethynyl-pyridin-2-ylamine Manufacturing Example 1-2-1 3-Iodopyridin-2-ylamine 50 s-12 2 N NH2 55 2 To a solution of 3-trimethylsilanylethynyl-pyridin-2- N NH2 ylamine (28.1 g, 148 mmol.) described in Manufacturing Example 1-2-2 in tetrahydrofuran (300 mL) was added tet A mixture of N-(3-Iodopyridin-2-yl)-2,2-dimethyl-propi rabutylammonium fluoride (1 Mtetrahydrofuran solution, 20 onamide (66.2 g, 218 mmol) described in Manufacturing 60 mL, 20 mmol), which was stirred for 15 minutes at room Example 39-1-2, 5 N aqueous sodium hydroxide solution temperature. Water was added to the reaction solution, which (200 mL) and methanol (200 mL) was stirred under reflux for was then extracted with ethyl acetate 4 times. The organic 1 hour and 20 minutes. The reaction solution was allowed to layer was dried over anhydrous sodium Sulfate, and the Sol room temperature and partitioned into water and ethyl vent was evaporated under a reduced pressure. The residue acetate. The aqueous layer was extracted with ethyl acetate 65 was purified by Silica gel chromatography (heptane:ethyl three times. The organic layers were combined, washed with acetate=1:1 then 1:2) to obtain the title compound (16.4g, saturated aqueous sodium chloride, and dried over anhydrous 93.7%). US 8,158,657 B2 97 98 H-NMR Spectrum (DMSO-d) & (ppm): 4.43 (1H, s), drous magnesium sulfate, and the solvent was evaporated 6.14 (2H, brs), 6.53 (1H, dd, J–4.8 Hz, 7.2 Hz), 7.53 (1H, d, under a reduced pressure. The residue was purified by NH J=7.2 Hz), 7.96 (1H, d, J=4.8 Hz). silica gel column chromatography (heptane:ethyl acetate-2: 1) to obtain the title compound (120 mg, 26%). Manufacturing Example 1-3-1 H-NMR Spectrum (CDC1) & (ppm): 4.08 (2H, s), 5.37 3-Trimethylsilanylethynyl-pyridin-2-ylamine (2H, s), 6.33 (1H, s), 6.45 (2H, brs), 6.79-6.82 (2H, m), 6.88-6.91 (1H, m), 7.30 (2H, d, J=8.1 Hz), 7.45 (2H, d, J=8.1 (Alternative Method) Hz), 7.57-7.61 (1H, m), 7.85 (1H, d, =7.3 Hz), 8.03 (1H, d, 10 J=5.5 Hz), 8.17 (1H, m). The starting material; (4-(pyridin-2-yloxymethyl)-phe nyl)-acetohydroximoyl chloride, was synthesized as follows. Si N 2 N 15 Manufacturing Example 2-1-1

r 2 (4-(Pyridin-2-yloxymethyl)-phenyl)methanol N NH2

To a solution of 2-amino-3-bromopyridine (5.72 g, 33.1 mmol) in N-methylpyrrolidinone (120 mL) were added trim Ne ethylsilyl acetylene (9.36 mL, 66.2 mmol), tetrakis(triph enylphosphine)palladium (0) (1.91 g, 1.66 mmol), copper(I) o-au iodide (630 mg, 3.31 mmol) and N,N-diisopropylethylamine 25 (11.5 mL, 66.2 mmol) at room temperature, which was stirred HO under nitrogen atmosphere for 6 hours at 70° C. Water was added to the reaction solution, which was then extracted with To a mixture of 1,4-benzenedimethanol (5.5 g, 40 mmol), ethyl acetate. The organic layer was washed with water and 2-fluoropyridine (1.3 g, 13 mmol) and N,N-dimethylforma saturated aqueous sodium chloride, and dried over anhydrous 30 mide (15 mL) was added sodium hydride (1.4g, 40 mmol. magnesium Sulfate, and the solvent was evaporated under a 66% in oil) at 0, which was stirred for 20 minutes at room reduced pressure. The residue was purified by silica gel col temperature and for 1 hour at 70° C. Water was added to the umn chromatography (heptane:ethyl acetate 2:1) to obtain reaction mixture, which was then extracted with ethyl acetate. the title compound (5.94 g., 94%). The organic layer was washed with Saturated aqueous sodium H-NMR Spectrum (DMSO-d) & (ppm): 0.23 (9H, s), 35 chloride, and the solvent was evaporated under a reduced 6.07 (2H, brs), 6.51 (1H, dd, J=49, 7.5 Hz), 7.49 (1H, dd, pressure. The residue was purified by NH silica gel column J=1.8, 7.5 Hz), 7.94 (1H, dd, J=1.8, 4.9 Hz). chromatography (ethyl acetate:heptane-1:1) to obtain the Example 2 title compound (1.9 g, 66%). 40 'H-NMR Spectrum (CDC1) & (ppm): 4.71 (2H, s), 5.38 3-(3-(4-(Pyridin-2-yloxymethyl)-benzyl)-isoxazol-5- (2H, s), 6.81 (1H, td, J=0.9, 8.4 Hz), 6.89 (1H, ddd, J=0.9, 5.1, yl)-pyridin-2-ylamine 7.1 Hz), 7.37-7.47 (4H, m), 7.59 (1H, ddd, J=2.0, 7.1,8.3 Hz), 8.17 (1H, ddd, J=0.7, 2.0, 5.1 Hz). 45 Manufacturing Example 2-1-2

4-(Pyridin-2-yloxymethyl)-benzaldehyde

50

Na o-au 55

To a solution of (4-(pyridin-2-yloxymethyl)-phenyl)-ac etohydroximoyl chloride (510 mg, 1.84 mmol) described in To a mixture of (4-(pyridin-2-yloxymethyl)-phenyl) Manufacturing Example 2-1-5 and 3-ethynyl-pyridin-2- 60 methanol (1.9 g, 8.6 mmol) described in Manufacturing ylamine (150 mg, 1.27 mmol) described in Manufacturing Example 2-1-1 and methylene chloride (30 mL) was added Example 1-2-3 in tetrahydrofuran (5 mL) was added triethy manganese dioxide (15 g, 17 mmol) at room temperature, lamine (708 ul, 5.08 mmol) at room temperature, which was which was stirred overnight at that temperature. The reaction stirred for 95 minutes at room temperature. Water was added mixture was filtered through a Celite pad, and the solvent was at room temperature to the reaction Solution, which was then 65 evaporated under a reduced pressure. The residue was puri extracted with ethyl acetate. The organic layer was washed fied by silica gel column chromatography (ethyl acetate:hep with Saturated aqueous sodium chloride, and dried over anhy tane=1:4) to obtain the title compound (770 mg, 42%). US 8,158,657 B2 99 100 H-NMR Spectrum (CDC1) & (ppm): 5.48 (2H, s), 6.85 Manufacturing Example 2-1-5 (1H, d, J=8.2 Hz), 6.90-6.93 (1H, m), 7.60-7.64 (3H, m), 7.89 (2H, d, J–8.1 Hz), 8.16 (1H, dd, J=1.3, 4.9 Hz), 10.0 (1H, s). 4-(Pyridin-2-yloxymethyl)-phenyl-acetohydroximoyl chloride Manufacturing Example 2-1-3

2-(4-((E)-2-Nitro-vinyl)-benzyloxy)-pyridine

10

y --Or-O 15 Lithium wire (323 mg, 46.6 mmol) was added to and dis solved in methanol (75 mL). To the mixture solution was added 2-(4-(2-nitro-ethyl)-benzyloxy)-pyridine (6.0 g, 23.3 A mixture of 4-(pyridin-2-yloxymethyl)-benzaldehyde mmol) described in Manufacturing Example 2-1-4. The reac (23.4 g 110 mmol) described in Manufacturing Example tion Solution was concentrated under a reduced pressure. 2-1-2, nitromethane (33.6 g. 550 mmol), ammonium acetate Toluene was added to the residue, and the solvent was con (17.0g, 220 mmol) and acetic acid (200 mL) was stirred for centrated under a reduced pressure. A solution of the resulting 1 hour and 45 minutes at 100° C. The reaction solution was 25 residue in methylene chloride (90 mL) and tetrahydrofuran stirred on an ice bath while adding a small amount of water, (45 mL) was cooled to -78°C., and titanium (IV) chloride and the precipitated solids were filtered to obtain the title (8.15 mL, 74.4 mmol) was added while stirring. Immediately compound (21.0 g, 74.5%). after addition of the titanium (IV) chloride, the reaction mix ture was stirred for 10 minutes at 0°C. and 30 minutes at room H-NMR Spectrum (DMSO-d) & (ppm): 5.41 (2H, s), 30 temperature. The reaction solution was poured into an ice 6.91 (1H, dd, J=0.8, 8.4 Hz), 6.99-7.10 (1H, m), 7.53 (2H, d, water and extracted with ethyl acetate. The organic layer was J=8.0 Hz), 7.72-7.79 (1H, m), 7.86 (2H, d, J=8.0 Hz), 8.13 dried over anhydrous magnesium sulfate, and the magnesium (1H, d, J=10 Hz), 8.15-8.20 (1H, m), 8.23 (1H, d, J=10 Hz). sulfate was removed by filtration. The filtrate was passed through a glass filter covered with neutral silica gel, washed 35 Manufacturing Example 2-1-4 with ethyl acetate. The resulting eluate was concentrated under a reduced pressure. A small amount of ethyl acetate was added to the residue, and the precipitated solids were filtered 2-(4-(2-Nitro-ethyl)-benzyloxy)-pyridine out to obtain the title compound (1.86 g. 28.8%). 40 H-NMR Spectrum (DMSO-d) & (ppm): 3.82 (2H, s), 5.33 (2H, s), 6.84-6.89 (1H, m), 6.97-7.01 (1H, m), 7.25 (2H, d, J=8.4 Hz), 7.41 (2H, d. J=8.4 Hz), 7.70-7.76 (1H, m), O N2 8.15-8.18 (1H, m), 11.7 (1H, s).

y o-au 45 Example 3 O 3-(3-(4-(6-Methyl-pyridin-2-yloxymethyl)-benzyl)- To a solution of 2-(4-((E)-2-nitro-vinyl)-benzyloxy)-pyri isoxazol-5-yl)-pyridin-2-ylamine dine (21.0 g, 81.9 mmol) described in Manufacturing 50 Example 2-1-3, acetic acid (21 mL) in dimethyl sulfoxide (200 mL) was added sodium borohydride (4.96g, 131 mmol) at room temperature while cooling appropriately. After addi tion of sodium borohydride, the cooling bath was removed, followed by stirring for 15 minutes at room temperature. The 55 reaction solution was partitioned into water and ethyl acetate. The ethyl acetate layer was washed with water twice and with saturated aqueous sodium chloride once, and dried over anhy drous magnesium sulfate, and the solvent was evaporated under a reduced pressure. The residue was purified by NH 60 silica gel column chromatography (ethyl acetate: heptane-1: 3) to obtain the title compound (16.3 g, 77.1%). H-NMR Spectrum (DMSO-d) & (ppm): 3.23 (2H, t, J=6.8 Hz), 4.85 (2H, t, J=6.8 Hz), 5.32 (2H, s), 6.82-6.88 (1H, 65 To a solution of 3-ethynyl-pyridin-2-ylamine (30 mg, 0.25 m), 6.96-7.01 (1H, m), 7.28 (2H, d, J=8.0 Hz), 7.38 (2H, d, mmol) described in Manufacturing Example 1-2-3 in anhy J=8.0 Hz), 7.69-7.74 (1H, m), 8.15-8.19 (1H, m). drous tetrahydrofuran (5 mL) was added (4-(6-methyl-pyri US 8,158,657 B2 101 102 din-2-yloxymethyl)-phenyl)-acetohydroximoyl chloride To a solution of 2-(4-bromo-benzyloxy)-6-methyl-pyri (222 mg, 0.76 mmol) described in Manufacturing Example dine (7.30 g, 26.2 mmol) described in Manufacturing 3-1-5 at room temperature. Triethylamine (142 uL. 1.0 mmol) Example 3-1-1 in anhydrous tetrahydrofuran (200 mL) was was added dropwise to the reaction solution, and stirred over night at room temperature. The reaction mixture was parti added dropwise n-butyl lithium (2.67 M n-hexane solution, tioned into water and ethyl acetate. The organic layer was 11.8 mL, 31.4 mmol) on a dry ice-ethanol bath (-78° C.), washed with water and Saturated aqueous sodium chloride, which was stirred for 30 minutes at -78° C. N.N-dimethyl and dried over anhydrous magnesium Sulfate, and the solvent formamide (4.04 mL, 52.4 mmol) was added to this mixture was evaporated under a reduced pressure. The residue was purified by NH silica gel column chromatography (ethyl at -78°C., and stirred for 5 minutes. Water and ethyl acetate acetate:heptane=1:3 then 1:1) to obtain the title compound 10 were added to the reaction mixture, which was stirred for 10 (10.5 mg, 11%). minutes at room temperature, and the organic layer was then H-NMR Spectrum (DMSO-d) & (ppm): 2.39 (3H, s), separated. This organic layer was washed with water and 4.04 (2H, s), 5.29 (2H, s), 6.26 (2H, brs), 6.61-6.64 (1H, m), saturated aqueous Sodium chloride, and dried over anhydrous 6.68-6.71 (1H, m), 6.81 (1H, s), 6.83 (1H, d. J=7.2 Hz), 7.33 (2H, d, J=8.0 Hz), 7.42 (2H, d, J=8.0 Hz), 7.57-7.61 (1H, dd, 15 magnesium sulfate, and the solvent was evaporated under a J=7.2, 8.4 Hz), 7.87 (1H, dd, J=2.0, 7.6 Hz), 8.08 (1H, dd, reduced pressure. The residue was purified by NH silica gel J=2.4, 5.0 Hz). column chromatography (ethyl acetate:heptane-1:3) to The starting material, (4-(6-methyl-pyridin-2-yloxym obtain the title compound (4.19 g, 70%). ethyl)-benzene)-acetohydroximoyl chloride, was synthe sized as follows. 'H-NMR Spectrum (CDC1) & (ppm): 2.44 (3H, s), 5.46 (2H, s), 6.12-6.64 (1H, m), 6.74-6.75 (1H, m), 7.44-7.50 (1H, Manufacturing Example 3-1-1 m), 7.62 (2H, d, J=8.0 Hz), 7.88 (2H, d, J=8.0 Hz), 10.0 (1H, S). 2-(4-Bromo-benzyloxy)-6-methyl-pyridine 25 Manufacturing Example 3-1-3

Br 2-Methyl-6-(4-((E)-2-nitro-vinyl)-benzyloxy)-pyri dine 30

To a solution of (4-bromo-phenyl)-methanol (4.54 g. 24.3 35 mmol) in N,N-dimethylformamide (50 mL) was added sodium hydride (999 mg, 25 mmol. 60% in oil) under nitro gen atmosphere on an ice bath (0°C.), which was stirred for 30 minutes at room temperature. 2-Fluoro-6-methylpyridine N (1.8 g. 16.2 mmol) was then added to the reaction mixture on 40 an ice bath (0°C.), and stirred for 5 hours at room tempera ture. The reaction mixture was partitioned into water and ethyl acetate on the ice bath (0° C.). The organic layer was washed with water and Saturated aqueous sodium chloride, and dried over anhydrous magnesium Sulfate, and the solvent 45 was evaporated under a reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate: heptane=1:15) to obtain the title compound (3.65 g, 81%). H-NMR Spectrum (CDC1) & (ppm): 2.44 (3H, s), 5.32 (2H, s), 6.57-6.59 (1H, m), 6.71-6.74 (1H, m), 7.26-7.35 (2H, m), 7.44-7.49 (3H m). 50 Manufacturing Example 3-1-2 To a solution of 4-(6-methyl-pyridin-2-yloxymethyl)-ben Zaldehyde (4.19 g, 18.5 mmol) described in Manufacturing 4-(6-Methyl-pyridin-2-yloxymethyl)-benzaldehyde Example 3-1-2 in acetic acid (30 mL) were added 55 nitromethane (5.65 g, 92.6 mmol) and ammonium acetate (2.85g. 37.0 mmol) under nitrogen atmosphere, which was stirred for 3 hours at 110°C. The reaction mixture was par titioned into water and ethyl acetate. The organic layer was 60 washed with water and Saturated aqueous sodium chloride, and dried over anhydrous magnesium Sulfate, and the solvent was evaporated under a reduced pressure to obtain the title compound (5.50 g) as a crude product. H-NMR Spectrum (CDC1) & (ppm): 2.45 (3H, s), 5.43 65 (2H, s), 6.05-6.28 (1H, m), 6.74-6.76 (1H, m), 7.47-7.51 (1H, m), 7.55 (4H, s), 7.59 (1H, d, J=13.6 Hz), 8.01 (1H, d, J=13.6 Hz). US 8,158,657 B2 103 104 Manufacturing Example 3-1-4 mixture was then stirred for further 60 minutes at room tem perature. Water, ethyl acetate and tetrahydrofuran were added to the reaction mixture on an ice bath (0°C.), and the organic 2-Methyl-6-(4-(2-nitro-ethyl)-benzyloxy)pyridine layer was separated. This organic layer was washed with water and Saturated aqueous sodium chloride, and dried over anhydrous magnesium Sulfate, and the solvent was evapo rated under a reduced pressure to obtain the title compound (484 mg.91%) as a crude product. O H-NMR Spectrum (DMSO-d) & (ppm): 2.42 (3H, s), 10 3.82 (2H, s), 5.33 (2H, s), 6.76 (1H, d, J–7.6 Hz), 6.92 (1H, d, O-1 J=7.6 Hz), 7.27 (2H, d, J=8.0 Hz), 7.44 (2H, d, J=8.0 Hz), 7.70 (1H, t, J=7.6 Hz), 11.8 (1H, brs). O N n Example 4 15 21 3-(3-(4-Butoxymethyl-benzyl)-isoxazol-5-yl)-pyri din-2-ylamine

To a solution of 2-methyl-6-(4-((E)-2-nitro-vinyl)-benzy loxy)-pyridine (5.00 g, 18.5 mmol) described in Manufactur ing Example 3-1-3 and acetic acid (5 mL) in dimethyl sul foxide (50 mL) was added sodium borohydride (1.2g, 29.6 mmol) under nitrogen atmosphere at room temperature while cooling appropriately, which was stirred for 10 minutes at 25 room temperature. Water was then added dropwise. The mix ture was partitioned into water and ethyl acetate. The organic layer was washed with water and Saturated aqueous sodium chloride, and dried over anhydrous magnesium sulfate, and the solvent was evaporated under a reduced pressure. The 30 residue was purified by NH silica gel column chromatogra phy (ethyl acetate: heptane=1:5 then 1:2) to obtain the title To a solution of 4-butoxymethyl-phenyl-acetohydroxi compound (2.8 g. 56%). moyl chloride (28 mg, 0.11 mmol) described in Manufactur 'H-NMR Spectrum (DMSO-d) & (ppm): 2.39 (3H, s), 35 ing Example 4-1-4 and 3-ethynyl-pyridin-2-ylamine (13 mg, 3.22 (2H, t, J=6.8 Hz), 4.85 (2H, t, J=6.8 Hz), 5.28 (2H, s), 0.11 mmol) described in Manufacturing Example 1-2-3 in 6.64 (1H, d, J=8.0 Hz), 7.84 (1H, d. J=8.0 Hz), 7.28 (2H, d, tetrahydrofuran (1 mL) was added triethylamine (31 uL, 0.22 mmol) at room temperature, which was stirred for 70 minutes J=7.6 Hz), 7.39 (2H, d, J=7.6 Hz), 7.59 (1H, t, J=8.0 Hz). at room temperature. The reaction solution was partitioned 40 into water and ethyl acetate at room temperature. The organic Manufacturing Example 3-1-5 layer was washed with Saturated aqueous Sodium chloride and dried over anhydrous magnesium Sulfate, and the solvent was evaporated under a reduced pressure. The residue was 4-(6-Methyl-pyridin-2-yloxymethyl)-phenyl)aceto purified by NH Silica gel column chromatography (heptane: hydroximoyl chloride 45 ethyl acetate=2:1) and then further purified by reverse-phase high performance liquid chromatography (using an acetoni trile-water mobile phase containing 0.1% trifluoroacetic acid) to obtain the title compound (2.3 mg, 5%) as a trifluo C roacetic acid salt. 50 MS m/e (ESI) (MH) 338.14 (MH) N O N The starting material, 4-butoxymethyl-phenyl-acetohy H-O1 n droximoyl chloride, was synthesized as follows. 2 Manufacturing Example 4-1-1 55 1-Bromo-4-butoxymethyl-benzene To a solution of 2-methyl-6-(4-(2-nitro-ethyl)-benzyloxy) pyridine (500 mg, 1.84 mmol) described in Manufacturing Example 3-1-4 in methanol (10 mL) was added lithium meth oxide (140 mg, 3.68 mmol) under nitrogen atmosphere at 60 Br room temperature, which was stirred for 30 minutes at room temperature. The reaction mixture was concentrated under a reduced pressure. Anhydrous methylene chloride (10 mL) N-1N1 and anhydrous tetrahydrofuran (5 mL) were added to the residue. Titanium (IV) chloride (667 uL, 6.07 mmol) was 65 To a solution of 4-bromobenzyl alcohol (10.0 g, 53.5 added dropwise to the reaction mixture on a dry ice-ethanol mmol) in N,N-dimethylformamide (200 mL) was added bath (-78°C.), and stirred for 45 minutes at 0°C. The reaction sodium hydride (3.08 g. 64.2 mmol. 50% in oil) at 0°C. This US 8,158,657 B2 105 106 mixture was stirred for 5 minutes at 0°C., and 1-bromobutane thereto and stirred for further 25 minutes. This reaction solu (7.47 mL, 69.3 mmol) was added thereto at 0°C. This mixture tion was partitioned into saturated aqueous sodium chloride was stirred for 40 minutes at room temperature and then and ethyl acetate at 0°C. The organic layer was washed with stirred for 25 minutes at 70° C. The reaction solution was saturated aqueous Sodium chloride and dried over anhydrous partitioned into water and ethyl acetate at 0°C. The organic magnesium sulfate, and the solvent was evaporated under a layer was washed with water and Saturated aqueous sodium reduced pressure. To a solution of the resulting residue in chloride, and dried over anhydrous magnesium sulfate, and dimethyl sulfoxide (100 mL) and acetic acid (6 mL) was the solvent was evaporated under a reduced pressure. The added sodium borohydride (1.84g, 46.1 mmol) at room tem residue was purified by silica gel column chromatography perature while cooling appropriately. This solution was then (heptane:ethyl acetate=20:1) to obtain the title compound 10 stirred for 80 minutes at room temperature. The reaction (11.5g, 89%). solution was partitioned into water and ethyl acetate. The H-NMR Spectrum (CDC1) & (ppm): 0.919 (3H, t, J–7.3 organic layer was washed with Saturated aqueous sodium Hz), 1.35-144 (2H, m), 1.56-1.63 (2H, m), 3.46 (2H, t, J–6.6 chloride and dried over anhydrous magnesium Sulfate, and Hz), 4.45 (2H, s), 7.21 (2H, d, J–8.1 Hz), 7.45-748 (2H, m). the solvent was evaporated under a reduced pressure. The 15 residue was purified by silica gel column chromatography Manufacturing Example 4-1-2 (heptane:ethyl acetate=4:1) to obtain the title compound 4-Butoxymethyl-benzaldehyde (2.68 g, 29%). 'H-NMR Spectrum (CDC1) & (ppm): 0.918 (3H, t, J–7.3 Hz), 1.37-1.42 (2H, m), 1.56-1.63 (2H, m), 3.31 (2H, t, J–7.3 HZ), 3.47 (2H, t, J=6.6 ha), 4.47 (2H, s), 4.60 (2H, t, J–7.3 Hz), 7.18 (2H, d, J=8.2 Hz), 7.30 (2H, d, J=8.2 Hz).

25 Manufacturing Example 4-1-4

N-1S-1 4-Butoxymethyl-phenyl-acetohydroximoyl chloride To a solution of 1-bromo-4-butoxymethyl-benzene (11.5 g, 47.3 mmol) described in Manufacturing Example 4-1-1 in 30 tetrahydrofuran (200 mL) was added n-butyl lithium (32.5 mL, 1.6 M hexane solution, 52.0 mmol) at -78° C. This mixture was stirred for 55 minutes at -78° C., and N,N- dimethylformamide (4.4 mL, 56.8 mmol) was added thereto at -78°C. This mixture was warmed to room temperature, 35 and stirred for 20 minutes. The reaction solution was parti tioned into water and ethyl acetate at 0°C. The organic layer was washed with Saturated aqueous Sodium chloride and dried over anhydrous magnesium Sulfate, and the solvent was evaporated under a reduced pressure. The residue was puri 40 fied by silica gel column chromatography (heptane:ethyl acetate=10:1) to obtain the title compound (7.39 g, 81%). 'H-NMR Spectrum (CDC1) & (ppm): 0.936 (3H, t, J–7.3 Hz), 1.37-1.47 (2H, m), 1.60- 1.67 (2H, m), 3.52 (2H, t, J–6.6 Hz), 4.58 (2H, s), 7.51 (2H, d, J=7.9 Hz), 7.86 (2H, m), 10.0 45 (1H, s). To a solution of 1-butoxymethyl-4-(2-nitroethyl)-benzene Manufacturing Example 4-1-3 (55 mg, 0.23 mmol) described in Manufacturing Example 4-1-3 in methanol (2 mL) was added sodium methoxide (1.49 1-Butoxymethyl-4-(2-nitroethyl)-benzene 50 M methanol solution, 47.3 uL, 0.23 mmol) at 0°C. The reaction Solution was stirred for 35 minutes at room tempera ture, and concentrated under a reduced pressure. To a solution of the residue in methylene chloride (2 mL) was added tita nium(IV) chloride (28 uL, 0.23 mmol) under nitrogen atmo 55 sphere at -78°C., which was then stirred for 30 minutes at 0° C. The reaction solution was partitioned into water and ethyl acetate at 0°C. The organic layer was washed with saturated 9N-1N1 aqueous Sodium chloride and dried over anhydrous magne sium Sulfate, and the magnesium Sulfate was removed by 60 To a solution of 4-butoxymethyl-benzaldehyde (7.39 g, filtration. The organic layer was filtered with silica gel, and 3.84 mmol) described in Manufacturing Example 4-1-2 in the filtrate was evaporated under a reduced pressure to obtain methanol (140 mL) was added nitromethane (2.70 mL, 49.9 the title compound (59 mg, 99%) as a crude product. mmol) followed by sodium methoxide (1.49 M methanol H-NMR Spectrum (CDC1) & (ppm): 0.90-0.94 (3H, m), solution, 9.41 mL, 46.1 mmol) at 0°C. The reaction solution 65 1.36-144 (2H, m), 1.56-1.64 (2H, m), 3.46-3.49 (2H, m), was stirred for 30 minutes at room temperature, and 5 N 3.79 (2H, s), 4.50 (2H, s), 7.23-7.26 (2H, m), 7.30-7.34 (2H, aqueous hydrochloric acid solution (120 mL) was added m), 8.29 (1H, s). US 8,158,657 B2 107 108 Example 5 H-NMR Spectrum (DMSO-d) & (ppm): 3.90 (2H, s), 6.25 (2H, brs), 6.68-6.72 (3H, m), 6.76 (1H, s), 7.11 (2H, d, 3-(3-(4-(2-Fluoro-benzyloxy)-benzyl)-isoxazol-5- J=8.6 Hz), 7.87 (1H, dd, J=1.5, 7.7 Hz), 8.10 (1H, brs), 9.29 yl)-pyridin-2-ylamine (1H, s). 5 Example 6

3-(3-(4-(3-Fluoro-benzyloxy)-benzyl)-isoxazol-5- 10 yl)-pyridin-2-ylamine

15

To a mixture of 4-(5-(2-amino-pyridin-3-yl)isoxazol-3-yl methyl)-phenol (4.2 mg, 0.016 mmol) described in Manufac turing Example 5-1-1 and methanol (0.4 mL) was added 1 N aqueous Sodium hydroxide Solution (16 LL, 0.016 mmol). This mixture was concentrated under a reduced pressure. To a mixture of the residue and N,N-dimethylformamide (0.5 25 mL) was added 2-fluorobenzyl bromide (2.3 uL, 0.019 mmol), which was stirred for 1 hour at room temperature. The To a mixture of 4-(5-(2-amino-pyridin-3-yl)isoxazol-3-yl reaction mixture was then purified directly by reverse-phase methyl)-phenol (4.2 mg, 0.016 mmol) described in Manufac turing Example 5-1-1 and methanol (0.4 mL) was added 1 N high performance liquid chromatography (using an acetoni 30 trile-water mobile phase containing 0.1% trifluoroacetic aqueous Sodium hydroxide Solution (16 LL, 0.016 mmol), acid) to obtain the title compound (3.3 mg, 43%) as a trifluo which was then concentrated under a reduced pressure. To a roacetic acid salt. mixture of the residue and N,N-dimethylformamide (0.5 mL) MS m/e (ESI) 376.14 (MH) was added 3-fluorobenzyl bromide (2.3 uL, 0.019 mmol), The starting material, 4-(5-(2-amino-pyridin-3-yl)isoX which was stirred for 1 hour at room temperature. The reac azol-3-ylmethyl)-phenol, was synthesized as follows. 35 tion mixture was purified directly by reverse-phase high per formance liquid chromatography (using an acetonitrile-water Manufacturing Example 5-1-1 mobile phase containing 0.1% trifluoroacetic acid) to obtain the title compound (4.3 mg, 55%) as a trifluoroacetic acid salt. 4-(5-(2-Amino-pyridin-3-yl)isoxazol-3-ylmethyl) phenol 40 MS m/e (ESI) 376.12 (MH) Example 7

45 3-(3-(4-(4-Fluoro-benzyloxy)-benzyl)-isoxazol-5- yl)-pyridin-2-ylamine

50

55 To a mixture of 3-(3-(4-benzyloxy-benzyl)-isoxazol-5-yl)- pyridin-2-ylamine (32 mg, 0.090 mmol) described in Example 1 and trifluoroacetic acid (1 mL) was added thio anisole (45 mg, 0.36 mmol) at room temperature, which was stirred for 2 hours at the same temperature. To a mixture of 60 saturated aqueous sodium hydrogencarbonate Solution and ethyl acetate was added the reaction mixture. The organic layer was separated and washed with Saturated aqueous To a mixture of 4-(5-(2-amino-pyridin-3-yl)isoxazol-3-yl Sodium chloride, and the solvent was evaporated under a methyl)-phenol (4.2 mg, 0.016 mmol) described in Manufac reduced pressure. The residue was purified by silica gel col 65 turing Example 5-1-1 and methanol (0.4 mL) was added 1 umn chromatography (ethyl acetate:heptane 4:1) to obtain Laqueous Sodium hydroxide solution (16 uL, 0.016 mmol), the title compound (24 mg, 100%). which was then concentrated under a reduced pressure. To a US 8,158,657 B2 109 110 mixture of the residue and N,N-dimethylformamide (0.5 mL) turing Example 5-1-1 and methanol (0.4 mL) was added 1 N was added 4-fluorobenzyl bromide (2.3 uL, 0.019 mmol), aqueous Sodium hydroxide Solution (16 LL, 0.016 mmol), which was stirred for 1 hour at room temperature. The reac which was then concentrated under a reduced pressure. To a tion mixture was purified as is by reverse-phase high perfor mixture of the residue and N,N-dimethylformamide (0.5 mL) mance liquid chromatography (using an acetonitrile-water was added 2-picolyl chloride (3.1 mg 0.019 mmol), which mobile phase containing 0.1% trifluoroacetic acid) to obtain was stirred for 2 hours at room temperature. The reaction the title compound (3.1 mg,39%) as a trifluoroacetic acid salt. mixture was purified as is by reverse-phase high performance MS m/e (ESI) 376.12 (MH) liquid chromatography (using an acetonitrile-water mobile phase containing 0.1% trifluoroacetic acid) to obtain the title Example 8 10 compound (3.6 mg, 39%) as a ditrifuoracetic acid salt. MS m/e (ESI) 359.16 (MH) 3-(3-(4-Cyclopropylmethoxy-benzyl)-isoxazol-5-yl)- pyridin-2-ylamine Example 10 15 3-(3-(4-(6-Methyl-pyridin-2-ylmethoxy)-benzyl)- isoxazol-5-yl)-pyridin-2-ylamine

25

To a mixture of 4-(5-(2-amino-pyridin-3-yl)isoxazol-3-yl methyl)-phenol (4.2 mg, 0.016 mmol) described in Manufac 30 turing Example 5-1-1 and methanol (0.4 mL) was added 1 N aqueous sodium hydroxide solution (16 ul, 0.016 Methanol (3 mL) and 1 Naqueous sodium hydroxide solu mmol), which was then concentrated under a reduced pres tion (0.18 mL) were added to 4-(5-(2-amino-pyridin-3-yl) sure. To a mixture of the residue and N,N-dimethylforma isoxazol-3-ylmethyl)-phenol (50 mg 0.19 mmol) described mide (0.5 mL) were added cyclopropylmethyl bromide (2.3 35 in Manufacturing Example 5-1-1, which was then dissolved uL, 0.019 mmol) and sodium iodide (1 mg, 7 mol) at room by irradiating ultrasonic wave. This solution was concen temperature, which was stirred for 2 hours at 60° C. The trated under a reduced pressure. To the resulting residue were reaction mixture was cooled to room temperature and then added 2-chloromethyl-6-methyl-pyridine (31.8 mg, 0.22 purified as is by reverse-phase high performance liquid chro mmol) described in Manufacturing Example 10-1-1 and N,N- matography (using an acetonitrile-water mobile phase con 40 dimethylformamide (2 mL), which was stirred for 20 minutes taining 0.1% trifluoroacetic acid). The eluate was neutralized at 60°C. The reaction solution was partitioned into water and by triethylamine while being concentrated. The solvent was ethyl acetate. The organic layer was separated, and the solvent evaporated under a reduced pressure. The residue was washed was evaporated under a reduced pressure. The residue was with water to obtain the title compound (1.6 mg, 30%). purified by NH Silica gel column chromatography (heptane: MS m/e (ESI) 322.19 (MH) 45 ethyl acetate=1:1) to obtain the title compound (36 mg, 51.7%). Example 9 H-NMR Spectrum (DMSO-d) & (ppm): 2.48 (3H, s), 3.96 (2H, s), 5.10 (2H, s), 6.25 (2H, brs), 6.69 (1H, dd, J=4.8, 3-(3-(4-(Pyridin-2-ylmethoxy)-benzyl)-isoxazol-5- 8.0 Hz), 6.79 (1H, s), 6.97 (2H, d, J=8.0 Hz), 7.18 (1H, d, yl)-pyridin-2-ylamine 50 J=7.6 Hz), 7.25 (2H, d, J=8.0 Hz), 7.27 (1H, d, J=7.6 Hz), 7.70 (1H, dd, J=7.6, 7.6Hz), 7.86 (1H, d, J=8.0Hz),8.08 (1H, d, J=4.8 Hz). The starting material, 2-chloromethyl-6-methyl-pyridine, was obtained as follows. 55 Manufacturing Example 10-1-1 2-Chloromethyl-6-methyl-pyridine

60

N

C 2 65 To a mixture of 4-(5-(2-amino-pyridin-3-yl)isoxazol-3-yl methyl)-phenol (4.2 mg, 0.016 mmol) described in Manufac US 8,158,657 B2 111 112 A Solution of (6-methyl-pyridin-2-yl)-methanol (1.44 g, Manufacturing Example 11-1-1 11.7 mmol), thionyl chloride (1.45 mL, 19.9 mmol) and methylene chloride (20 mL) was stirred under reflux for 40 minutes. The reaction solution was cooled to room tempera 2,4-Dimethyl-pyridine 1-oxide ture and then concentrated under a reduced pressure. The residue was partitioned into sodium bicarbonate solution and diethyl ether. The organic layer was concentrated under a reduced pressure, and the residue was purified by Silica gel chromatography (ethyl acetate) to obtain the title compound (1.42g, 85.8%). 10 'H-NMR Spectrum (DMSO-d) & (ppm): 2.47 (3H, s), 4.72 (2H, s), 7.22 (1H, d, J=7.6 Hz), 7.33 (1H, d, J=7.6 Hz), N 7.72 (1H, dd, J=7.6, 7.6 Hz). 2 Example 11 15 O 3-(3-(4-(4-Methyl-pyridin-2-ylmethoxy)-benzyl)- isoxazol-5-yl)-pyriin-2-ylamine

To a solution of 2.4-lutidine (2.0 g. 18.7 mmol) in methyl ene chloride (100 mL) was added 3-chloroperoxybenzoic acid (5.07 g. 29.4 mmol), which was stirred for 20 minutes at 25 room temperature. A small amount of Saturated aqueous Sodium hydrogen Sulfite solution was added to the reaction Solution, and the organic layer was separated after vigorous stirring. This organic layer was washed with 5 N aqueous

sodium hydroxide solution (5.9 mL), and dried over anhy 30 drous magnesium Sulfate. The solvent was evaporated under a reduced pressure to obtain the title compound (1.54 g. 66.9%). The title compound was used in the following reac tion without being purified.

35 Manufacturing Example 11-1-2

Acetic acid 4-methyl-pyridin-2-ylmethyl ester 40

45 Methanol (3 mL) and 1 Naqueous sodium hydroxide solu tion (0.18 mL) were added to 4-(5-(2-amino-pyridine-3-yl) isoxazole-3-ylmethyl)-phenol (50 mg, 0.19 mmol) described in Manufacturing Example 5-1-1, which was then dissolved by irradiating ultrasonic wave. This solution was concen 50 trated under a reduced pressure. To the resulting residue were added 2-chloromethyl-4-methyl-pyridine (31.8 mg, 0.22 mmol) described in Manufacturing Example 11-1-4 and N,N- dimethylformamide (2 mL), which was stirred for 10 minutes at 60°C. The reaction solution was partitioned into water and 55 ethyl acetate. The organic layer was separated, and the solvent Acetic anhydride (30 mL) was added to 2,4-dimethyl-py was evaporated under a reduced pressure. The residue was ridine 1-oxide (1.93 g, 15.7 mmol) described in Manufactur purified by NH silica gel column chromatography (heptane: ing Example 11-1-1, and the mixture was stirred for 10 min ethyl acetate=1:1) to obtain the title compound (21 mg, utes at 110° C. The reaction solution was allowed to room 30.2%). 60 temperature and concentrated under a reduced pressure. The 'H-NMR Spectrum (DMSO-d) & (ppm): 2.33 (3H, s), resulting residue was purified by silica gel column chroma 3.96 (2H, s), 5.11 (2H, s), 6.25 (2H, brs), 6.69 (1H, dd, J=4.8, 8.0 Hz), 6.79 (1H, s), 6.98 (2H, d, J=8.4 Hz), 7.17 (1H, d, tography (heptane:ethyl acetate-1:2, then ethyl acetate) to J=7.6 Hz), 7.25 (2H, d, J=8.4 Hz), 7.34 (1H, s), 7.87 (1H, d, obtain the title compound (774 mg, 29.8%). J=7.6 Hz), 8.09 (1H, d. J–4.8 Hz), 8.41 (1H, d, J–4.8 Hz). 65 'H-NMR Spectrum (DMSO-d) & (ppm): 2.11 (3H, s), The starting material, 2-chloromethyl-4-methyl-pyridine, 2.32 (3H, s), 5.09 (2H, s), 7.16 (1H, d, J=5.2 Hz), 7.23 (1H, s), was synthesized as follows. 8.39 (1H, d, J=5.2 Hz). US 8,158,657 B2 113 114 Manufacturing Example 11-1-3 Example 12 3-(3-(6-Benzyloxy-pyridin-3-ylmethyl)-isoxazol-5- (4-Methyl-pyridin-2-yl)-methanol yl)-pyridin-2-ylamine

10 N

HO 2 N 15

5 N Aqueous sodium hydroxide solution (2 mL) and To a solution of 3-ethynyl-pyridin-2-ylamine (400 mg. methanol (4 mL) were added to acetic acid 4-methyl-pyridin 3.39 mmol) described in Manufacturing Example 1-2-3 in 2-ylmethyl ester (774 mg. 4.69 mmol) described in Manufac anhydrous tetrahydrofuran (20 mL) was added (2-benzyloxy turing Example 11-1-2, and this mixture was stirred for 10 pyridin-5-yl)-acetohydroximoyl chloride (2.50 g, 9.03 minutes at 60°C. The reaction solution was partitioned into mmol) under nitrogen atmosphere at room temperature. Tri water and ethyl acetate. The separated aqueous layer was 25 ethylamine (1.89 mL, 13.6 mmol) was then added dropwise further extracted with ethyl acetate twice. The ethyl acetate thereto, and stirred for 1.5 hours at room temperature. The layers were combined and dried over anhydrous magnesium reaction mixture was partitioned into water and ethyl acetate Sulfate, and the solvent was evaporated under a reduced pres at room temperature. The organic layer was washed with sure to obtain the title compound (410 mg, 71.0%). water and Saturated aqueous sodium chloride, and dried over 'H-NMR Spectrum (DMSO-d) & (ppm): 2.32 (3H, s), 30 anhydrous magnesium Sulfate, and the solvent was evapo 4.52 (2H, brs), 5.35 (1H, brs), 7.06 (1H, d, J=5.2 Hz), 7.29 rated under a reduced pressure. The residue was purified by (1H, s), 8.32 (1H, d, J=5.2 Hz). NH silica gel column chromatography (ethyl acetate:hep tane=1:3 then 1:2) to obtain the title compound (315 mg. 26%). Manufacturing Example 11-1-4 35 'H-NMR Spectrum (DMSO-d) & (ppm): 4.00 (2H, s), 5.34 (2H, s), 6.27 (2H, brs), 6.70 (1H, dd, J–4.8, 7.6Hz), 6.84 2-Chloromethyl-4-methyl-pyridine (1H, s), 6.86 (1H, d, J=8.8 Hz), 7.31-744 (5H, m), 7.69 (1H, dd, J–2.4, 8.4 Hz), 7.87 (1H, dd, J=2.0, 7.4 Hz), 8.09 (1H, dd, J=2.4, 4.8 Hz), 8.17 (1H, d, J=2.4 Hz). 40 The starting material. (2-benzyloxy-pyridin-5-yl)-aceto hydroximoyl chloride, was synthesized as follows. Manufacturing Example 12-1-1 45 2-Benzyloxy-5-bromopyridine N

C 2 N Br 50

A mixture solution of (4-methyl-pyridine-2-yl)-methanol (410 mg, 3.33 mmol) described in Manufacturing Example 55 11-1-3, thionyl chloride (0.49 mL, 6.66 mmol) and methylene To a solution of phenyl-methanol (20.5 g., 190 mmol) in chloride (10 mL) was stirred under reflux for 5 minutes. The N,N-dimethylformamide (200 mL) was added sodium reaction Solution was allowed to room temperature and con hydride (7.6 g., 190 mmol) under nitrogen atmosphere on an centrated under a reduced pressure. The resulting residue was 60 ice bath (0° C.), which was stirred for 30 minutes at room partitioned into diethyl ether and saturated sodium bicarbon temperature. 2,5-Dibromopyridine was then added thereto on ate Solution. The organic layer was purified by silica gel the ice bath (0°C.), and stirred for 60 minutes at room tem column chromatography (ethyl acetate) to obtain the title perature. The reaction mixture was partitioned into water and compound (340 mg, 72.1%). ethyl acetate on the ice bath (0° C.). The organic layer was H-NMR Spectrum (DMSO-d) & (ppm): 2.37 (3H, s), 65 washed with water and Saturated aqueous sodium chloride, 4.72 (2H, s), 7.20 (1H, d. J–5.2 Hz), 7.38 (1H, s), 8.40 (1H, d, and dried over anhydrous magnesium Sulfate, and the solvent J=5.2 Hz). was evaporated under a reduced pressure. The residue was US 8,158,657 B2 115 116 purified by silica gel column chromatography (ethyl acetate: was evaporated under a reduced pressure to obtain the title heptane=1:20 then 1:10) to obtain the title compound (15.1 g, compound (5.60 g, 96%) as a crude product. 90%). 'H-NMR Spectrum (DMSO-d) & (ppm): 5.43 (2H, s), H-NMR Spectrum (CDC1) & (ppm): 5.34 (2H, s), 6.71 7.01 (1H, d, J=8.8 Hz), 7.34-7.47 (5H, m), 8.16 (1H, d, J=13.6 6.73 (1H, m), 7.32-7.45 (5H, m), 7.64-7.67 (1H, m), 8.20- 5 Hz), 8.24 (1H, d, J=13.6 Hz), 8.27 (1H, dd, J=2.4, 8.8 Hz), 8.21 (1H, m). 8.64 (1H, d. J=2.4 Hz). Manufacturing Example 12-1-2 Manufacturing Example 12-1-4 6-Benzyloxy-pyridin-3-carbaldehyde 10 2-Benzyloxy-5-(2-nitro-ethyl)pyridine

O 15 H N

r2 2 N O

To a solution of 2-benzyloxy-5-bromopyridine (15.1 g, To a solution of 2-benzyloxy-5-((E)-2-nitro-vinyl)-pyri 57.0 mmol) described in Manufacturing Example 12-1-1 in dine (5.80 g, 22.8 mmol) described in Manufacturing anhydrous tetrahydrofuran (250 mL) were added dropwise 25 Example 12-1-3 and acetic acid (5.80 mL) in dimethyl sul n-butyl lithium (2.67 M n-hexane solution, 25.6 mL, 68.4 foxide (70 mL) was added sodium borohydride (1.44g, 36.2 mmol) under nitrogen atmosphere on a dry ice-ethanol bath mmol) under nitrogen atmosphere at room temperature while (-78°C.), which was stirred for 30 minutes at -78° C. N.N- cooling appropriately, which was stirred for 10 minutes at Dimethylformamide (6.60 mL. 85.5 mmol) was then added 30 room temperature. The reaction mixture was partitioned into thereto at -78°C., and stirred for 30 minutes. Water and ethyl water and ethyl acetate. The organic layer was washed with acetate were added to the reaction mixture, and the organic water and saturated aqueous sodium chloride, and dried over layer was separated after stirring for 10 minutes at room anhydrous magnesium Sulfate, and the solvent was evapo temperature. The organic layer was washed with water and rated under a reduced pressure. The residue was purified by saturated aqueous sodium chloride, and dried over anhydrous 35 NH silica gel column chromatography (ethyl acetate:hep magnesium Sulfate, and the solvent was evaporated under a tane=1:4) to obtain the title compound (2.50 g, 43%). reduced pressure. The residue was purified by NH silica gel H-NMR Spectrum (DMSO-d) & (ppm): 3.17 (2H, t, column chromatography (ethyl acetate:heptane=1:7 then 1:5) J=6.8 Hz), 4.84 (2H, d, J=6.8 Hz), 5.31 (2H, s), 6.84 (1H, d, to obtain the title compound (4.87 g, 40%). J=8.4 Hz), 7.31-7.42 (5H, m), 7.68 (1H, dd, J=2.4, 8.4 Hz), H-NMR Spectrum (CDC1) & (ppm): 5.49 (2H, s), 6.89- 40 8.06 (1H, d. J=2.4 Hz). 6.92 (1H, m), 7.34-7.48 (5H, m), 8.07-8.10 (1H, m), 8.64 Manufacturing Example 12-1-5 8.65 (1H, m), 9.97 (1H, s). Manufacturing Example 12-1-3 (2-Benzyloxy-pyridin-5-yl)-acetohydroximoyl chloride 2-Benzyloxy-5-((E)-2-nitro-vinyl)-pyridine 45

50 o-1 N N21 N 2 N O 55 To a solution of 2-benzyloxy-5-(2-nitro-ethyl)pyridine (3.97 g. 15.4 mmol) described in Manufacturing Example 12-1-4 in methanol (25 mL) was added lithium methoxide To a solution of 6-benzyloxy-pyridin-3-carbaldehyde (1.17 g., 30.8 mmol) under nitrogen atmosphere at room tem (4.87 g. 22.8 mmol) described in Manufacturing Example 60 perature, which was stirred at room temperature for 30 min 12-1-2 in acetic acid (30 mL) were added nitromethane (6.96 utes. The reaction mixture was concentrated under a reduced g, 114 mmol) and ammonium acetate (3.51 g, 45.6 mmol) pressure. Anhydrous methylene chloride (30 mL) and anhy under nitrogen atmosphere at room temperature, which was drous tetrahydrofuran (20 mL) were added to the residue. stirred for 2.5 hours at 110° C. The reaction mixture was Titanium (IV) chloride (5.42 mL, 49.3 mmol) was added partitioned into water and ethyl acetate. The organic layer was 65 dropwise into the reaction mixture on a dry ice-ethanol bath washed with water and Saturated aqueous sodium chloride, (-78° C.), and stirred for 45 minutes at 0° C. Water, ethyl and dried over anhydrous magnesium Sulfate, and the solvent acetate and tetrahydrofuran were added to the reaction mix US 8,158,657 B2 117 118 ture on an ice bath (0° C.), and the organic layer was sepa 445 mmol) was added in one portion while stirring at room rated. The organic layer was washed with water and Saturated temperature. The reaction solution was stirring for 10 minutes aqueous Sodium chloride, and dried over anhydrous magne at room temperature. Water (3.5 L) was added to the reaction sium Sulfate, and the solvent was evaporated under a reduced solution, and the precipitated solids were filtered out. The pressure to obtain the title compound (3.4g, 80%) as a crude resulting aqueous layer was extracted with ethyl acetate (1.3 product. L) 3 times. The ethyl acetate layers were combined and the H-NMR Spectrum (DMSO-d) & (ppm): 3.79 (2H, s), Solvent was evaporated under a reduced pressure. The residue 5.34 (2H, s), 6.87 (1H, d, J=8.4 Hz), 7.30-7.62 (5H, m), 7.61 was purified by Silica gel chromatography (heptane:ethyl (1H, dd, J=2.4, 8.4 Hz), 7.08 (1H, d, J=2.4 Hz), 11.8 (1H, s). acetate=2:3) to obtain the title compound (23.8 g. 22.8%). 10 'H-NMR Spectrum (DMSO-d) & (ppm): 5.41 (2H, brs), Example 13 5.57 (1H, d, J=8.0Hz), 5.64 (2H, brs), 7.37 (1H, d, J=8.0 Hz). 3-(3-(4-Benzyloxy-benzyl)-isoxazol-5-yl)-pyridin-2, Manufacturing Example 13-1-2 6-diamine 15 3-Trimethylsilanylethynyl-pyridin-2,6-diamine

25 HN N

To a mixture of 3-iodo-pyridin-2,6-diamine (20.0 g. 85.2 mmol) described in Manufacturing Example 13-1-1, trimeth 30 ylsilyl acetylene (24.2 mL, 170 mmol), copper (I) iodide To a mixture of 4-benzyloxy-phenyl-acetohydroximoyl (3.25 g, 17.0 mmol) N,N-diisopropylethylamine (19.1 g, 148 chloride (140 mg, 0.51 mmol) described in Manufacturing mmol) and N-methylpyrrolidinone (286 mL) was added tet Example 1-1-3 and tetrahydrofuran (10 mL) were added rakis(triphenylphosphine)palladium (O) (9.81 g, 8.52 mmol) 3-ethynyl-pyridin-2,6-diamine (102 mg, 0.76 mmol) under argon atmosphere, which was stirred for 30 minutes at described in Manufacturing Example 13-1-3 and triethy 35 room temperature. The reaction solution was partitioned into lamine (0.71 mL, 5.1 mmol), which was stirred overnight at water and ethyl acetate. The ethyl acetate layer was washed room temperature. The reaction mixture was then stirred for with water 4 times and dried over sodium sulfate, and the further 1.5 hours at 55°C. The reaction solution was cooled to Solvent was evaporated under a reduced pressure. The residue room temperature and concentrated under a reduced pressure. was purified by NH silica gel chromatography (heptane:ethyl The residue was filtered by NH silica gel column chromatog 40 acetate-4:1 then 1:1). The solids obtained by concentrating raphy (ethyl acetate) to obtain a crude product. The crude the eluate under a reduced pressure were washed with heptane product was purified by reverse-phase high performance liq containing a small amount of ethyl acetate to obtain the title uid chromatography (using an acetonitrile-water mobile compound (10.5 g. 60.0%). phase containing 0.1% trifluoroacetic acid). The solvent was H-NMR Spectrum (DMSO-d) & (ppm): 0.20 (9H, s), evaporated under a reduced pressure, and the residue was 45 5.53 (2H, brs), 5.66 (1H, d, J=8.0 Hz), 5.95 (2H, brs), 7.11 filtered with NH silica gel to obtain the title compound (51 (1H, d, J=8.0 Hz). mg, 27%). H-NMR Spectrum (DMSO-d) & (ppm): 3.87 (2H, s), Manufacturing Example 13-1-3 5.07 (2H, s), 5.79 (2H, brs), 5.82 (1H, d, J=8.6 Hz), 6.10 (2H, brs), 6.34 (1H, s), 6.94-6.98 (2H, m), 7.20-7.24 (2H, m), 3-Ethynyl-pyridin-2,6-diamine 7.30-7.45 (5H, m), 7.51 (1H, d, J=8.4 Hz). 50 The starting material, 3-ethynyl-pyridin-2,6-diamine, was synthesized as follows. Manufacturing Example 13-1-1 2 55 su1 3-Iodo-pyridin-2,6-diamine 2 HN N NH2

60 To a solution of 3-trimethylsilanylethynyl-pyridin-2,6-di amine (7.0 g, 34.1 mmol.) described in Manufacturing Example 13-1-2 in tetrahydrofuran (100 mL) was added tet 2 rabutylammonium fluoride (1Mtetrahydrofuran solution, 17 HN N NH2 mL, 17 mmol) on an ice bath, which was stirred for 10 65 minutes at room temperature. Water was added to the reaction 2,6-Diaminopyridine (100 g, 916 mmol) was dissolved in solution, which was then extracted with ethyl acetate 3 times. dimethylsulfoxide (400 mL), and N-iodosuccinimide (100 g, The extract was dried over sodium sulfate, and the solvent US 8,158,657 B2 119 120 was evaporated under a reduced pressure. The residue was To a solution of 3-ethynyl-pyridin-2,6-diamine (300 mg. purified by silica gel chromatography (ethylacetate) to obtain 2.25 mmol) described in Manufacturing Example 13-1-3 in the title compound (3.35 g, 73.8%). anhydrous tetrahydrofuran (30 mL) was added (4-(6-methyl H-NMR Spectrum (DMSO-d) & (ppm): 4.08 (1H, s), pyridin-2-yloxymethyl)-phenyl)-acetohydroximoyl chloride 5.57 (2H, brs), 5.68 (1H, d, J=8.0 Hz), 5.89 (2H, brs), 7.14 5 (1.50 g, 5.16 mmol) described in Manufacturing Example (1H, d, J=8.0 Hz). 3-1-5 under nitrogen atmosphere at room temperature. Tri ethylamine (1.25 mL, 9.00 mmol) was then added dropwise at Example 14 room temperature, and stirred for 1.5 hours at room tempera ture. Water and ethyl acetate were added to the reaction mix 3-(3-(4-Pyridin-2-yloxymethyl)-benzyl)-isoxazol-5- 10 ture at room temperature, and the organic layer was separated. yl)-pyridine-2,6-diamine The organic layer was washed with water and Saturated aque ous sodium chloride, and dried over anhydrous magnesium Sulfate, and the solvent was evaporated under a reduced pres sure. The residue was purified by NH silica gel column chro 15 matography (ethyl acetate:heptane=2:1) to obtain the title compound (637 mg, 73%). 'H-NMR Spectrum (DMSO-d) & (ppm): 2.39 (3H, s), 3.96 (2H, s), 5.29 (2H, s), 5.80 (2H, brs), 5.83 (1H, d, J=8.8 Hz), 6.11 (2H, brs), 6.37 (1H, s), 6.63 (1H, dd, J=0.4, 8.2 Hz), 6.83 (1H, dd, J=0.4, 7.4 Hz), 7.31 (2H, d, J=8.0Hz), 7.41 (2H, d, J=8.4 Hz), 7.51 (1H, d, J=8.4 Hz), 7.58 (1H, t, J=8.0 Hz). Example 16 25

To a solution of 3-ethynyl-pyridin-2,6-diamine (120 mg. 3-(3-(4-Butoxymethyl-benzyl)-isoxazol-5-yl)-pyri 0.90 mmol) described in Manufacturing Example 13-1-3 and din-2,6-diamine 4-(pyridine-2-yloxymethyl)-phenyl-acetohydroximoyl chlo ride (390 mg, 1.41 mmol) described in Manufacturing 30 Example 2-1-5 in tetrahydrofuran (5.0 mL) was added tri ethylamine (502 uL, 3.6 mmol) at 0°C. The reaction mixture was stirred for 1 hour and 30 minutes at room temperature.

The mixture was partitioned into ethyl acetate and water. The 35 organic layer was separated, washed with water and Saturated aqueous Sodium chloride, and dried over anhydrous magne sium Sulfate, and the solvent was evaporated under a reduced pressure. The residue was purified by silica gel column chro matography (heptane:ethyl acetate-1:1, then ethyl acetate) to 40 obtain the title compound (290 mg, 86.2%). 'H-NMR Spectrum (DMSO-d) & (ppm): 3.95 (2H, s), 5.31 (2H, s), 5.79 (2H, brs), 5.82 (1H, d, J=8.4 Hz), 6.11 (2H, brs), 6.37 (1H, s), 6.84-6.86 (1H, m), 6.97-7.00 (1H, m), 7.31 (2H, d, J=8.2 Hz), 7.39 (2H, d, J=8.2 Hz), 7.51 (1H, d, J=8.4 45 Hz), 7.69-7.73 (1H, m), 8.16-8.18 (1H, m). Example 15

3-(3-(4-(6-Methyl-pyridin-2-yloxymethyl)-benzyl)- 50 To a solution of 3-ethynyl-pyridin-2,6-diamine (14.6 mg, isoxazol-5-yl)-pyridin-2,6-diamine 0.11 mmol) described in Manufacturing Example 13-1-3 and 4-butoxymethyl-phenyl-acetohydroximoyl chloride (28 mg, 0.11 mmol) described in Manufacturing Example 4-1-4 in

55 tetrahydrofuran was added triethylamine (31 uL, 0.22 mmol), which was stirred for 4 hours at room temperature. The reac tion solution was partitioned into water and ethyl acetate at room temperature. The organic layer was washed with Satu rated aqueous sodium chloride and dried over anhydrous 60 magnesium sulfate, and the solvent was evaporated under a reduced pressure. The residue was purified by NH silica gel column chromatography (heptane:ethyl acetate-2:1) and then purified again by reverse-phase high performance liquid chromatography (using an acetonitrile-water mobile phase 65 containing 0.1% trifluoroacetic acid) to obtain the title com pound (6.7 mg, 13%) as a trifluoroacetic acid salt. MS m/e (ESI) 353.34 (MH) US 8,158,657 B2 121 122 Example 17 and the solids were dried azeotropically with toluene to obtain the title compound (1.17 g, 55%). 3-(3-(4-Phenoxy-benzyl)-isoxazol-5-yl)-pyridin-2,6- H-NMR Spectrum (DMSO-d) & (ppm): 5.38 (1H, m), diamine 5.73 (1H, d, J=5.2 Hz), 6.58 (1H, d, J=44 Hz), 6.91-7.00 (4H, m), 7.09-7.13 (1H, m), 7.34-7.39 (4H, m).

Manufacturing Example 17-1-2

10 1-((E)-2-Nitro-vinyl)-4-phenoxy-benzene

15

N

To a solution of 3-ethynyl-pyridin-2,6-diamine (170 mg. 28 mmol) described in Manufacturing Example 13-1-3 in anhydrous tetrahydrofuran (10 mL) was added (4-phenoxy benzene)-acetohydroximoyl chloride (652 mg, 2.49 mmol) described in Manufacturing Example 17-1-4 under nitrogen 25 A solution of Sodium 2-nitro-1-(4-phenoxy-phenyl)-etha atmosphere at room temperature. Triethylamine (714 uL. nolate (1.17 g. 4.16 mmol) described in Manufacturing 5.12 mmol) was then added dropwise, and stirred for 1 hour at Example 17-1-1, acetic anhydride (510 mg, 4.99 mmol) and room temperature. The reaction mixture was partitioned into triethylamine (696 uL, 4.99 mmol) in anhydrous tetrahydro water and ethyl acetate at room temperature. The organic furan (20 mL) was stirred overnight under nitrogen atmo layer was washed with water and Saturated aqueous sodium 30 chloride, and dried over anhydrous magnesium sulfate, and sphere at room temperature. The reaction mixture was parti the solvent was evaporated under a reduced pressure. The tioned into water and ethyl acetate at room temperature. The residue was purified by NH silica gel column chromatogra organic layer was washed with water and Saturated aqueous phy (ethyl acetate: heptane=1:2 then 2:1) to obtain the title Sodium chloride, and dried over anhydrous magnesium Sul compound (314 mg. 68%). 35 fate, and the solvent was evaporated under a reduced pressure to obtain the title compound (1.4g, 70%, purity: ca. 50%) as 'H-NMR Spectrum (DMSO-d) & (ppm): 4.00 (2H, s), a crude product. 4.74 (2H, brs), 5.50 (2H, brs), 5.94 (1H, d, J=8.8 Hz), 6.03 (1H, s), 6.96-7.02 (2H, m), 7.08-7.12(1H, m), 7.22-7.26 (5H, m), 7.30-7.35 (1H, m), 7.52 (1H, d, J=8.8 Hz). Manufacturing Example 17-1-3 The starting material, (4-phenoxy-benzene)-acetohydroxi 40 moyl chloride, was synthesized as follows. 1-(2-Nitro-ethyl)-4-phenoxy-benzene Manufacturing Example 17-1-1 45 Sodium 2-nitro-1-(4-phenoxy-phenyl)-ethanolate N

50 O OH

To a solution of 1-((E)-2-nitro-vinyl)-4-phenoxy-benzene (1.40g, 2.90 mmol, purity: 50%) described in Manufacturing 55 Example 17-1-2 in methanol (15 mL) was added sodium borohydride (274 mg, 7.25 mmol) at room temperature while To a solution of 4-phenoxybenzaldehyde (1.5 g, 7.56 cooling appropriately under nitrogen atmosphere, which was mmol) in methanol (12 mL) was added dropwise sodium stirred for 10 minutes at room temperature. Water was then methoxide (1.49 M methanol solution, 0.19 mL, 0.91 mmol) 60 added dropwise at room temperature while cooling appropri under nitrogen atmosphere at room temperature. ately. The reaction mixture was extracted with ethyl acetate. Nitromethane (530 uL. 9.84 mmol) was added dropwise to The organic layer was washed with water and Saturated aque the reaction solution on an ice bath (0°C.). Sodium methox ous sodium chloride, and dried over anhydrous magnesium ide (1.49 M methanol solution, 1.66 mL, 8.16 mmol) was Sulfate, and the solvent was evaporated under a reduced pres added dropwise thereto at room temperature, and the Solution 65 Sure. The residue was purified by Silica gel column chroma was stirred for 30 minutes at room temperature. The precipi tography (ethyl acetate:heptane=1:5) to obtain the title com tated solids were filtered and dried under reduced pressure, pound (199 mg, 28%). US 8,158,657 B2 123 124 H-NMR Spectrum (DMSO-d) & (ppm): 3.21 (2H, t, Manufacturing Example 18-1-1 in tetrahydrofuran (3 mL) J=6.8 Hz), 4.84 (2H, t, J=6.8 Hz), 6.94-7.00 (4H, m), 7.11 was added 5 Naqueous sodium hydroxide solution (51.2 LL. 7.15 (1H, m), 7.28-7.30 (2H, m), 7.36-7.40 (2H, m). 0.26 mmol), which was irradiated by ultrasonic wave for 5 minutes. Next, the reaction solution was concentrated under a Manufacturing Example 17-1-4 reduced pressure to obtain solids (77.9 mg). To a solution of the resulting solids (14.5 mg 0.05 mmol) in N,N-dimethyl (4-Phenoxy-benzene)-acetohydroximoyl chloride formamide (1 mL) was added 2-fluorobenzyl bromide (11.5 uL, 0.10 mmol), which was stirred for 2 hours at room tem perature. The reaction Solution was partitioned into water and 10 ethyl acetate. The organic layer was washed with water and saturated aqueous Sodium chloride, and dried over anhydrous magnesium sulfate, and the solvent was evaporated under a C reduced pressure. The residue was purified by silica gel col umn chromatography (ethyl acetate) and then further purified 15 by reverse-phase high performance liquid chromatography (using an acetonitrile-water mobile phase containing 0.1% trifluoroacetic acid) to obtain the title compound (6.7 mg, 36%) as a trifluoroacetic acid salt. H-NMR Spectrum (CDC1) & (ppm): 3.96 (2H, s), 4.57 To a solution of 1-(2-nitro-ethyl)-4-phenoxy-benzene (100 (2H, brs), 5.12 (2H, s), 5.90 (2H, brs), 5.91 (1H, d, J=8.4 Hz), mg, 0.41 mmol) described in Manufacturing Example 17-1-3 5.98 (1H, s), 6.95 (2H, d, J=8.4 Hz), 7.05-7.11 (1H, m), in methanol (3 mL) was added sodium methoxide (1.49 M 7.14-7.24 (1H, m), 7.20 (2H, d, J=8.4 Hz), 7.28-7.33 (1H, m), methanol Solution, 83.9L, 0.41 mmol) under nitrogen atmo 25 7.48 (1H, d, J=8.4 Hz), 7.45-7.51 (1H, ). sphere, which was stirred for 30 minutes at room temperature. MS m/e (ESI) 391.01 (MH) The reaction mixture was concentrated under a reduced pres sure. Anhydrous methylene chloride (3 mL) was added to the residue. Titanium (IV) chloride (54.2 LL, 0.49 mmol) was Manufacturing Example 18-1-1 added dropwise to the reaction mixture on an ice bath (0°C.), and stirred for 30 minutes at room temperature. Water, ethyl 30 acetate and tetrahydrofuran were added on the ice bath (0°C.) 4-(5-(2,6-Diamino-pyridine-3-yl)-isoxazol-3-ylm to partition the reaction mixture. The organic layer was ethyl)phenol washed with water and Saturated aqueous sodium chloride, and dried over anhydrous magnesium Sulfate, and the solvent was evaporated under a reduced pressure. The residue was 35

purified by silica gel column chromatography (ethyl acetate: heptane=1:5) to obtain the title compound (51 mg, 47%). H-NMR Spectrum (DMSO-d) & (ppm): 3.80 (2H, s), 6.96-7.03 (4H, m), 7.12-7.16 (1H, m), 7.26-7.28 (2H, m), 7.36-7.41 (2H, m), 11.7 (1H, s). 40 Example 18 3-(3-(4-(2-Fluoro-benzyloxy)-benzyl)-isoxazol-5- yl)-pyridin-2,6-diamine 45

50 To a solution of 3-(3-(4-benzyloxy-benzyl)-isoxazol-5- yl)-pyridin-2,6-diamine (100 mg 0.27 mmol) described in Example 13 in trifluoroacetic acid (3 mL) was added thioani sole (126 uL) at room temperature, which was stirred for 2 hours at room temperature. Saturated aqueous Sodium hydro 55 gen carbonate solution was added to the reaction Solution at 0° C., which was then extracted with ethyl acetate. The organic layer was washed with water and Saturated aqueous Sodium chloride, and dried over anhydrous magnesium Sul 60 fate, and the solvent was evaporated under a reduced pressure. The residue was purified by silica gel column chromatogra phy (ethyl acetate) to obtain the title compound (72.4 mg. 95%). H-NMR Spectrum (DMSO-d) & (ppm): 3.82 (2H, s), 65 5.79 (2H, brs), 5.83 (1H, d, J=8.4 Hz), 6.10 (2H, brs), 6.32 To a solution of 4-(5-(2,6-diamino-pyridin-3-yl)-isoxazol (1H, s), 6.70 (2H, d, J=8.4 Hz), 7.09 (2H, d, J=8.4 Hz), 7.51 3-ylmethyl)-phenol (72.4 mg., 0.26 mmol) described in (1H, d, J=8.4 Hz), 9.27 (1H, s). US 8,158,657 B2 125 126 Example 19 the resulting solids (13.7 mg 0.05 mmol) in N,N-dimethyl formamide (1 mL) was added 4-fluorobenzyl bromide (11.2 3-(3-(4-(3-Fluoro-benzyloxy)-benzyl)-isoxazol-5- uL, 0.09 mmol), which was stirred for 2.5 hours at room yl)-pyridin-2,6-diamine temperature. The reaction Solution was partitioned into water and ethyl acetate. The organic layer was washed with water and Saturated aqueous sodium chloride, and dried over anhy drous magnesium sulfate, and the solvent was evaporated under a reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate), the mixture was 10 purified by reverse-phase high performance liquid chroma tography (using an acetonitrile-water mobile phase contain ing 0.1% trifluoroacetic acid) and then purified by preparative thin-layer chromatography (ethyl acetate:hexane-1:1) to 15 obtain the title compound (4.0 mg, 18%). 'H-NMR Spectrum (CDC1-d) & (ppm): 3.96 (2H, s), 4.53 (2H, brs), 5.00 (2H, s), 5.30 (2H, brs), 5.91 (1H, d, J=8.0 Hz), 5.98 (1H, s), 6.92 (2H, dd, J=2.0, 6.8 Hz), 7.05-7.15 (2H, m), 7.20 (2H, d, J=8.4 Hz), 7.26-7.46 (2H, m), 7.48 (1H, d, J-8.0 To a solution of 4-(5-(2,6-diamino-pyridin-3-yl)-isoxazol Hz). 3-ylmethyl)-phenol (72.4 mg., 0.26 mmol) described in MS m/e (ESI) 391.04 (MH) Manufacturing Example 18-1-1 in tetrahydrofuran (3 mL) was added 5 Naqueous sodium hydroxide solution (51.2 LL. Example 21 0.26 mmol), which was irradiated by ultrasonic wave for 5 minutes. Next, the reaction solution was concentrated under a 25 reduced pressure to obtain solids (77.9 mg). To a solution of 3-(3-(4-Cyclopropylmethoxy-benzyl)-isoxazol-5-yl)- the resulting solids (11.3 mg, 0.04 mmol) in N,N-dimethyl pyridin-2,6-diamine formamide (1 mL) was added 3-fluorobenzyl bromide (9.1 uL, 0.07 mmol), which was stirred for 2 hours at room tem perature. The reaction Solution was partitioned into water and 30

ethyl acetate. The organic layer was washed with water and saturated aqueous sodium chloride, and dried over anhydrous magnesium Sulfate, and the solvent was evaporated under a reduced pressure. The residue was purified by silica gel col umn chromatography (ethyl acetate), and further purified by 35 reverse-phase high performance liquid chromatography (us ing an acetonitrile-water mobile phase containing 0.1% trif luoroacetic acid) to obtain the title compound (6.7 mg, 36%) as a trifluoroacetic acid salt. MS m/e (ESI) 391.34 (MH) 40 Example 20 3-(3-(4-(4-Fluoro-benzyloxy)-benzyl)-isoxazol-5- yl)-pyridin-2,6-diamine 45 To a solution of 4-(5-(2,6-diamino-pyridin-3-yl)-isoxazol 3-ylmethyl)-phenol (72.4 mg. 0.26 mmol) described in Manufacturing Example 18-1-1 in tetrahydrofuran (3 mL) was added 5 Naqueous sodium hydroxide solution (51.2 LL. 0.26 mmol), which was irradiated by ultrasonic wave for 5 50 minutes. Next, the reaction solution was concentrated under a reduced pressure to obtain solids (77.9 mg). To a solution of the resulting solids (8.3 mg, 0.03 mmol) in N,N-dimethylfor mamide (1 mL) was added cyclopropylmethyl bromide (5.3 uL, 0.06 mmol), which was stirred for 5 hours at room tem 55 perature. The mixture was purified by reverse-phase high performance liquid chromatography (using an acetonitrile water mobile phase containing 0.1% trifluoroacetic acid), and then further purified by preparative thin-layer chromatogra phy (ethyl acetate) to obtain the title compound (1.1 mg, 60 To a solution of 4-(5-(2,6-diamino-pyridin-3-yl)-isoxazol 12%). 3-ylmethyl)-phenol (72.4 mg., 0.26 mmol) described in 'H-NMR Spectrum (CDC1-d) & (ppm): 0.33-0.36 (2H, Manufacturing Example 18-1-1 in tetrahydrofuran (3 mL) m), 0.63-0.66 (2H, m), 1.24-1.29 (1H, s), 3.79 (2H, d, J–4.8 was added 5 Naqueous sodium hydroxide solution (51.2 LL. HZ), 3.96 (2H, s), 4.57 (2H, brs), 5.34 (2H, brs), 5.92 (1H, d, 0.26 mmol), which was irradiated by ultrasonic wave for 5 65 J=8.4 Hz), 5.99 (1H, s), 6.87 (2H, dd, J-2.0, 6.8 Hz), 7.19 minutes. Next, the reaction solution was concentrated under a (2H, dd, J=2.0, 6.8 Hz), 7.49 (1H, d, J=8.4 Hz). reduced pressure to obtain solids (77.9 mg). To a solution of MS m/e (ESI) 337.11 (MH) US 8,158,657 B2 127 128 Example 22 To 4-(5-(2,6-diamino-pyridin-3-yl)-isoxazol-3-ylmethyl)- phenol (150 mg, 0.53 mmol) described in Manufacturing 3-(3-(4-(Pyridin-2-ylmethoxy)-benzyl)-isoxazol-5- Example 18-1-1 were added methanol (3 mL) and 1 Naque yl)-pyridin-2,6-diamine ous sodium hydroxide solution (0.53 mL), which was then dissolved by irradiating ultrasonic wave. This solution was concentrated under a reduced pressure. To the resulting resi due was added 2-chloromethyl-6-methyl-pyridine (90.2 mg, 0.64 mol.) described in manufacturing Example 10-1-1 and N,N-dimethylformamide (2 mL), which was stirred for 2 10 hours and 50 minutes at 60° C. The reaction solution was partitioned into water and ethylacetate. The organic layer was evaporated under a reduced pressure. The residue was puri fied by NH silica gel column chromatography (heptane:ethyl 15 acetate=1:2, then ethyl acetate) to obtain the title compound (106 mg, 51.5%). H-NMR Spectrum (DMSO-d) & (ppm): 2.48 (3H, s), 3.88 (2H, s), 5.10 (2H, s), 5.78 (2H, brs), 5.82 (1H, d, J–8.4 To a solution of 4-(5-(2,6-diamino-pyridin-3-yl)-isoxazol Hz), 6.10 (2H, brs), 6.34 (1H, s), 6.96 (2H, d, J=8.0 Hz), 7.18 3-ylmethyl)-phenol (49.7 mg, 0.18 mmol) described in (1H, d, J=8.0 Hz), 7.22 (2H, d, J=8.0 Hz), 7.27 (1H, d, J=8.0 Manufacturing Example 18-1-1 in tetrahydrofuran (3 mL) hz), 7.50 (1H, d, J=8.4 Hz), 7.70 (1H, dd, J=8.0, 8.0 Hz). was added 5 Naqueous sodium hydroxide solution (35.2 LL. 0.18 mmol), which was irradiated by ultrasonic wave for 5 Example 24 minutes. Next, the reaction solution was concentrated under a reduced pressure to obtain solids (90.6 mg). The resulting 25 solids were made into an N,N-dimethylformamide (3 mL) 3-(3-(4-(4-Methyl-pyridin-2-ylmethoxy)-benzyl)- solution. Tetrahydrofuran (390 LL) and 1 Naqueous sodium isoxazol-5-yl)-pyridin-2,6-diamine hydroxide solution (390 uL, 0.39 mol) were added to 2-pico lyl chloride hydrochloride (50 mg. 0.39 mmol), and then the organic layer was separated to obtain tetrahydrofuran solu 30 tion of 2-picolyl chloride. A part of the solution (0.30 mL) was added to the aforementioned N,N-dimethylformamide solution, and stirred for 15 hours at room temperature. The

reaction solution was partitioned into water and ethyl acetate. The organic layer was washed with water and Saturated aque 35 ous sodium chloride, and dried over anhydrous magnesium Sulfate, and the solvent was evaporated under a reduced pres Sure. The residue was purified by Silica gel column chroma tography (ethyl acetate) to obtain the title compound (42.5 mg, 38%). 40 'H-NMR Spectrum (DMSO-d) & (ppm): 3.88 (2H, s), 5.15 (2H, s), 5.79 (2H, brs), 5.83 (1H, dd, J=1.2, 8.4 Hz), 6.11 (2H, brs), 6.35 (1H, s), 6.97 (2H, d, J=8.0 Hz), 7.22 (2H, d, J=8.4 Hz), 7.33 (1H, dd, J=5.2, 8.0 Hz), 7.49 (1H, d, J=8.0 Hz), 7.51 (1H, d, J=8.0), 7.82 (1H, dd, J=8.0, 8.0 Hz), 8.57 45 (1H, dd, J=0.8, 4.8 Hz). MS m/e (ESI) 374.28 (MH"). To 4-(5-(2,6-diamino-pyridin-3-yl)-isoxazol-3-ylmethyl)- phenol (80 mg 0.28 mmol) described in Manufacturing Example 23 Example 18-1-1 were added methanol (4 mL) and 1 Naque 50 ous sodium hydroxide solution (0.29 mL), which was then 3-(3-(4-(6-Methyl-pyridin-2-ylmethoxy)-benzyl)- dissolved by irradiating ultrasonic wave. This solution was isoxazol-5-yl)-pyridin-2,6-diamine concentrated under a reduced pressure. To the resulting resi due was added 2-chloromethyl-4-methyl-pyridine (50.9 mg, 0.36 mol.) described in Manufacturing Example 11-1-4 and 55

N,N-dimethylformamide (3 mL), which was stirred for 10 minutes at 60°C. The reaction solution was partitioned into water and ethyl acetate. The organic layer was evaporated under a reduced pressure. The residue was purified by NH silica gel column chromatography (heptane:ethyl acetate=1: 60 2, then ethyl acetate) to obtain the title compound (40 mg. 36.5%). 'H-NMR Spectrum (DMSO-d) & (ppm): 2.32 (3H, s), 3.88 (2H, s), 5.10 (2H, s), 5.79 (2H, brs), 5.82 (1H, d, J=8.4 65 Hz), 6.10 (2H, brs), 6.35 (1H, s), 6.97 (2H, d, J=8.0 Hz), 7.15 (1H, d, J=5.2 Hz), 7.22 (2H, d, J=8.0 Hz), 7.34 (1H, s), 7.50 (1H, d, J=8.4 Hz), 8.41 (1H, d, J=5.2 Hz). US 8,158,657 B2 129 130 Example 25 was then extracted with ethyl acetate-tetrahydrofuran (3:2). The organic layer was washed with Saturated aqueous sodium 3-(3-(6-Benzyloxy-pyridin-3-ylmethyl)-isoxazol-5- chloride, and the solvent was evaporated under a reduced yl)-pyridin-2,6-diamine pressure. The residue was purified by NH silica gel column chromatography (ethyl acetate: heptane=2:3) to obtain the title compound (8.8 mg, 41%). 'H-NMR Spectrum (CDC1) & (ppm): 3.47 (3H, s), 3.99 (2H, s), 4.42 (2H, s), 5.05 (2H, s), 5.50 (2H, brs), 6.23 (1H, s), 10 6.82(1H, d, J–7.9 Hz), 6.93-6.97 (2H, m), 7.18-7.22 (2H, m), 7.31-744 (5H, m), 7.72 (1H, d, J=7.7 Hz). The starting material, 3-ethynyl-6-methoxymethyl-pyri din-2-ylamine, was synthesized as follows.

15 Manufacturing Example 26-1-1

2-Amino-6-chloro-nicotinic acid To a solution of 3-ethynyl-pyridin-2,6-diamine (230 mg. 1.73 mmol) described in Manufacturing Example 13-1-3 in anhydrous tetrahydrofuran (20 mL) was added (2-benzyloxy pyridin-5-yl)-acetohydroximoyl chloride (1.00 g, 3.61 mmol) described in Manufacturing Example 12-1-5 under 25 nitrogen atmosphere at room temperature. Triethylamine N (965 uL, 6.92 mmol) was added dropwise to the mixture and OH stirred for 1.5 hours at room temperature. The reaction mix 2 ture was partitioned into water and ethyl acetate at room C N NH2 temperature. The organic layer was washed with water and 30 saturated aqueous sodium chloride, and dried over anhydrous magnesium sulfate, and the solvent was evaporated under a reduced pressure. The residue was purified by NH silica gel A mixture of 2,6-dichloro-nicotinic acid (31 g, 0.14 mol) column chromatography (ethyl acetate:heptane 2:1) to and 28% aqueous ammonia solution (200 mL) was stirred in obtain the title compound (470 mg, 73%). a sealed tube for 10 hours at 135° C. This reaction solution 'H-NMR Spectrum (DMSO-d) & (ppm): 3.92 (2H, s), 35 was cooled to room temperature, and the excess ammonia gas 5.33 (2H, s), 5.81 (2H, brs), 5.83 (1H, d, J=8.4 Hz), 6.11 (2H, was removed under a reduced pressure. Water was added to brs), 6.40 (1H, s), 6.85 (1H, d. J=8.8 Hz), 7.31-7.39 (3H, m), the residue to a total of 1000 mL, the mixture was cooled to 0° 742-744 (2H, m), 7.52 (1H, d. J=8.4 Hz), 7.66 (1H, dd, C., and citric acid was added to a pH being about 6. The J=2.4, 8.4 Hz), 8.14 (1H, d. J=2.4 Hz). 40 precipitated solids were filtered out to obtain the title com pound (12 g, 49%). Example 26 H-NMR Spectrum (DMSO-d) & (ppm): 6.63 (1H, d, 3-(3-(4-Benzyloxy-benzyl)-isoxazol-5-yl)-6-meth J=8.1 Hz), 7.55 (2H, brs), 8.02 (1H, d, J=8.1 Hz). oxymethyl-pyridin-2-ylamine 45 Manufacturing Example 26-1-2

2-Amino-6-chloro-nicotinic acid methyl ester 50

55

C N

To a mixture of (4-benzyloxy-phenyl)-acetohydroximoyl 60 Concentrated sulfuric acid (25 mL) and 2-amino-6-chloro chloride (19 mg, 0.069 mmol) described in Manufacturing nicotinic acid (4.3 g, 25 mmol) described in Manufacturing Example 1-1-3 and tetrahydrofuran (1 mL) were added Example 26-1-1 were added to methanol (50 mL) on an ice 3-ethynyl-6-methoxymethyl-pyridin-2-ylamine (8.6 mg, bath, and stirred at 70° C. for 5 hours. The reaction mixture 0.053 mmol) described in Manufacturing Example 26-1-7 was cooled and then neutralized by addition of aqueous and triethylamine (15 uL, 0.11 mmol) at room temperature, 65 Sodium hydrogen carbonate (90 g) solution. The precipitated which was stirred for 5.5 hours at room temperature. Water solids were filtered to obtain the title compound (3.2g, 17 was added at room temperature to the reaction mixture, which mmol. 68%). US 8,158,657 B2 131 132 H-NMR Spectrum (CDC1) & (ppm): 3.88 (3H, s), 6.62 H-NMR Spectrum (CDC1) & (ppm): 3.47 (3H, s), 3.88 (1H, d, J=8.2 Hz), 8.05 (1H, d, J=8.1 Hz). (3H, s), 4.41 (2H, s), 6.74 (1H, d, J–7.9 Hz), 8.14 (1H, d, J=7.9 Hz). Manufacturing Example 26-1-3 Manufacturing Example 26-1-5 Tributyl-methoxymethyl-stannane (2-Amino-6-methoxymethyl-pyridin-3-yl)methanol

10

r 15 N OH To a mixture of diisopropylamine (9.4 mL, 67 mmol) and tetrahydrofuran (150 mL) was added dropwise n-butyl lithium (2.4 Mn-hexane solution, 25 mL, 61 mmol) at -78D, which was stirred for 30 minutes at the same temperature. Tributyltin hydride (16 mL, 61 mmol) was added dropwise to the reaction mixture at the same temperature, and stirred for To a mixture of lithium aluminum hydride (80%, 220 mg. 30 minutes at 0°C. The reaction mixture was cooled to -78° 4.6 mmol) and tetrahydrofuran (5 mL) was added 2-amino C., and chloromethyl methyl ether (4.6 mL, 61 mmol) was 25 6-methoxymethyl-nicotinic acid methyl ester (300 mg, 1.5 added dropwise thereto. The reaction mixture was gradually mmol) described in Manufacturing Example 26-1-4 at 0°C., warmed to room temperature. Water was added to the reaction which was stirred for 20 minutes at the same temperature. An mixture, which was then extracted with diethyl ether. The aqueous 28% ammonia Solution was added dropwise to the organic layer was washed with Saturated aqueous sodium reaction mixture at 0°C. The mixture was warmed to room chloride, and the solvent was evaporated under a reduced 30 temperature and filtered. The filtrate was concentrated under pressure. The residue was purified by neutral silica gel col a reduced pressure to obtain the title compound (260 mg. umn chromatography (ethyl acetate:heptane=1:30) to obtain 100%). the title compound (18 g, 86%). 'H-NMR Spectrum (CDC1) & (ppm): 3.45 (3H, s), 4.39 H-NMR Spectrum (CDC1) & (ppm): 0.88-0.93 (15H, m), 35 (2H, s), 4.62 (2H, s), 5.03 (2H, brs), 6.70 (1H, d. J–7.3 Hz), 1.26-1.35 (6H, m), 1.47-1.55 (6H, m), 3.30 (3H, s), 3.71 (2H, 7.31 (1H, d, J=7.5 Hz). t, J=6.8 Hz). Manufacturing Example 26-1-4 Manufacturing Example 26-1-6 40 2-Amino-6-methoxymethyl-nicotinic acid methyl 2-Amino-6-methoxymethyl-pyridine-3-carbaldehyde ester

45 O N 1 N 50 O 2 1. N NH2

A mixture of 2-amino-6-chloro-nicotinic acid methyl ester (1.4g, 7.6 mmol) described in Manufacturing Example 26-1- 55 To a mixture of (2-amino-6-methoxymethyl-pyridin-3-yl) 2, tributyl-methoxymethyl-Stannane (3.1 g, 9.1 mmol) methanol (260 mg, 1.5 mmol) described in Manufacturing described in Manufacturing Example 26-1-3, tetrakis(triph Example 26-1-5 and methylene chloride (15 mL) was added enylphosphine)palladium (440 mg, 0.38 mmol) and N-meth manganese dioxide (1.3g, 15 mmol), which was stirred over ylpyrrolidinone (20 mL) was stirred for 3.5 hours at 130° C. night at room temperature. The reaction mixture was filtered The reaction mixture was cooled to room temperature, and 60 aqueous potassium fluoride Solution and ethyl acetate were through a Celite pad, and the filtrate was concentrated under added to the reaction mixture, which was then filtered through a reduced pressure. The residue was purified by silica gel a Celite pad. The organic layer was separated and washed column chromatography (ethyl acetate:heptane 3:2) to with Saturated aqueous sodium chloride, and the solvent was obtain the title compound (210 mg, 81%). evaporated under a reduced pressure. The residue was puri 65 'H-NMR Spectrum (CDC1) & (ppm): 3.48 (3H, s), 4.44 fied by silica gel column chromatography (ethyl acetate:hep (2H, s), 6.87 (1H, d, J=7.9 Hz), 7.82 (1H, d, J=7.7 Hz), 9.84 tane=1:2) to obtain the title compound (0.93 g. 63%). (1H, s). US 8,158,657 B2 133 134 Manufacturing Example 26-1-7 column chromatography (ethyl acetate:heptane 2:3) to obtain the title compound (10 mg, 48%). 3-Ethynyl-6-methoxymethyl-pyridin-2-ylamine 'H-NMR Spectrum (CDC1) & (ppm): 3.47 (3H, s), 4.07 (2H, s), 4.44 (2H, s), 5.37 (2H, s), 5.56 (2H, brs), 6.25 (1H, s), 6.79-6.84 (2H, m), 6.87-6.91 (1H, m), 7.30 (2H, d, J–7.9 Hz), 7.44 (2H, d, J=7.9 Hz), 7.57-7.61 (1H, m), 7.73 (1H, d, J=7.9 2 Hz), 8.18 (1H, d, J–4.2 Hz). N Example 28 10 -O 4. NH2 5-(3-(4-Benzyloxy-benzyl)-isoxazol-5-yl)-pyridin-2- ylamine To a mixture of diisopropylamine (0.15 mL, 1.1 mmol) and tetrahydrofuran (2 mL) was added dropwise n-butyl lithium 15

(1.6 M n-hexane solution, 0.68 mL, 1.1 mmol) at -78D, which was stirred for 30 minutes at that temperature. Trim ethylsilyl diazomethane (2 M n-hexane solution, 0.50 mL. 0.99 mmol) was added to the reaction mixture at -78°C., and stirred for 30 minutes at that temperature. A mixture of 2-amino-6-methoxymethyl-pyridine-3-carbaldehyde (150 mg, 0.90 mmol) described in Manufacturing Example 26-1-6 and tetrahydrofuran (1.5 mL) was added dropwise to the reaction mixture at -78°C., and stirred for 30 minutes at 0°C. The reaction mixture was cooled to -78°C., and a mixture of 25 acetic acid (0.10 mL) and tetrahydrofuran (1 mL) was added dropwise to the reaction mixture. This reaction mixture was gradually warmed to 0°C., and partitioned into water and To a solution of 5-ethynyl-pyridin-2-ylamine (10 mg. 85 ethyl acetate. The organic layer was washed with Saturated umol) described in Manufacturing Example 28-1-3 and aqueous Sodium chloride, and the solvent was evaporated 30 4-benzyloxy-phenyl-acetohydroximoyl chloride (70 mg. under a reduced pressure. The residue was purified by silica 0.25 mmol) described in Manufacturing Example 1-1-3 in gel column chromatography (ethyl acetate:heptane 2:3) to tetrahydrofuran (2 mL) was added triethylamine (35ul, 0.25 mmol) at room temperature, which was stirred for 3 hours and obtain the title compound (73 mg, 50%). 40 minutes at room temperature. The reaction solution was H-NMR Spectrum (CDC1) & (ppm): 3.40 (1H, s), 3.45 35 partitioned into water and ethyl acetate at 0°C. The organic (3H, s), 4.39 (2H, s), 5.07 (2H, brs), 6.72 (1H, d, J=7.7 Hz), layer was washed with Saturated aqueous Sodium chloride 7.58 (1H, d, J=7.5 Hz). and dried over anhydrous magnesium Sulfate, and the solvent was evaporated under a reduced pressure. The residue was Example 27 purified by reverse-phase high performance liquid chroma 6-Methoxymethyl-3-(3-(4-pyridin-2-yloxymethyl)- 40 tography (using an acetonitrile-water mobile phase contain benzyl)-isoxazol-5-yl)-pyridin-2-ylamine ing 0.1% trifluoroacetic acid) to obtain the title compound (1 mg, 3%) as a trifluoroacetic acid salt. MS m/e (ESI) 358.00 (MH) The starting material, 5-ethynyl-pyridin-2-ylamine, was 45 synthesized as follows. Manufacturing Example 28-1-1 2-Nitro-5-trimethylsilanylethynyl-pyridine 50

55 To a mixture of 4-(pyridin-2-yloxymethyl)-phenyl-aceto hydroximoyl chloride (18 mg 0.064 mmol) described in o, Manufacturing Example 2-1-5 and tetrahydrofuran (1 mL) N1 NN were added 3-ethynyl-6-methoxymethyl-pyridin-2-ylamine 60 (8.6 mg, 0.053 mmol) described in Manufacturing Example O 26-1-7 and triethylamine (15 uL, 0.11 mmol), which was stirred for 2 hours at room temperature. The reaction mixture To a solution of 5-bromo-2-nitropyridine (1.00 g, 4.93 was partitioned into water and ethyl acetate at room tempera mmol) in N-methylpyrrolidinone (20 mL) were added trim ture. The organic layer was washed with Saturated aqueous 65 ethylsilyl acetylene (1.39 mL, 9.85 mmol), tetrakis(triph Sodium chloride, and the solvent was evaporated under a enylphosphine)palladium (0) (114 mg. 985 umol), copper(I) reduced pressure. The residue was purified by NH silica gel iodide (37.5 mg, 197 umol) and N,N-diisopropylethylamine US 8,158,657 B2 135 136 (1.72 mL, 9.85 mmol) at room temperature, which was stirred H-NMR Spectrum (CDC1) & (ppm): 3.07 (1H, s), 4.73 under nitrogen atmosphere for 4 hours at 65°C. The reaction (2H, brs), 6.46 (1H, d, J=8.6Hz), 7.53 (1H, dd, J=2.2, 8.6Hz), solution was partitioned into water and ethyl acetate at 0°C. 8.21 (1H, s). The organic layer was washed with water and Saturated aque ous sodium chloride, and dried over anhydrous magnesium Example 29 Sulfate, and the solvent was concentrated under a reduced pressure. The residue was purified by silica gel column chro 3-(5-(4-Benzyloxy-benzyl)-isoxazol-3-yl)-pyridin-2- matography (heptane:ethyl acetate=6:1) to obtain the title ylamine compound (490 mg. 45%). H-NMR Spectrum (CDC1) & (ppm): 0.298 (9H, s), 8.03 10 8.05 (1H, m), 8.22 (1H, J=8.4 Hz), 8.66 (1H, d, J-2.0 Hz). Manufacturing Example 28-1-2 5-Trimethylsilanylethynyl-pyridin-2-ylamine 15 e N s /

2 N NH2

To a solution of 3-(5-(4-benzyloxy-benzyl)-isoxazol-3- 25 yl)-5-chloro-pyridin-2-ylamine (50 mg, 0.13 mmol) HN N described in Manufacturing Example 29-2-3 in N-methylpyr rolidinone (2 mL) were added formic acid (7.3 uL, 0.19 To a solution of 2-nitro-5-trimethylsilanylethynyl-pyridine mmol), N,N-diisopropylethylamine (67 uL, 0.38 mmol) and (405 mg, 1.84 mmol) described in Manufacturing Example tetrakis(triphenylphosphine)palladium (O) (15 mg, 13 mmol) 28-1-1 in tetrahydrofuran (10 mL) and water (5 mL) were 30 under nitrogen atmosphere at room temperature, which was added iron powder (514 mg., 9.21 mmol) and ammonium stirred for 2 hours and 20 minutes at 100° C. Water and ethyl chloride (197 mg, 3.69 mmol) at room temperature, which acetate were added to the reaction solution at room tempera was stirred for 75 minutes at 70° C. The reaction solution was ture, which was then filtered through a Celite pad. The filtrate cooled to room temperature and filtered through a Celite pad, was partitioned into water and ethylacetate. The organic layer 35 was washed with Saturated aqueous Sodium chloride and and the filtrate was concentrated under a reduced pressure. dried over anhydrous magnesium sulfate, and the solvent was The residue was purified by silica gel column chromatogra evaporated under a reduced pressure. The residue was puri phy (heptane:ethyl acetate=1:1) to obtain the title compound fied by reverse-phase high performance liquid chromatogra (319 mg, 91%). phy (using an acetonitrile-water mobile phase containing H-NMR Spectrum (CDC1) & (ppm): 0.237 (9H, s), 4.73 40 0.1% trifluoroacetic acid) and then purified again by NH (2H, brs), 6.44 (1H, d, J=8.6Hz), 7.51 (1H, dd, J=2.2, 8.4 Hz), silica gel column chromatography (heptane:ethyl acetate=1: 8.19 (1H, d. J=2.2 Hz). 1) to obtain the title compound (5 mg, 11%). Manufacturing Example 28-1-3 'H-NMR Spectrum (CDC1) & (ppm): 4.07 (2H, s), 5.07 (2H, s), 6.24 (1H, s), 6.34 (2H, brs), 6.67 (1H, dd, J–4.9, 7.5 5-Ethynyl-pyridin-2-ylamine 45 Hz), 6.95-6.98 (2H, m), 7.20-7.23 (2H, m), 7.31-7.45 (5H, m), 7.66 (1H, dd, J=1.7, 7.5 Hz), 8.11 (1H, dd, J=1.7, 4.9 Hz). MS m/e (ESI) 358.20 (MH) The starting material, 3-(5-(4-benzyloxy-benzyl)-isox aZol-3-yl)-5-chloro-pyridin-2-ylamine, was synthesized as 2 st-1 50 follows. Manufacturing Example 29-1-1 2 HN N 2-Amino-pyridine-3-carbaldehyde oxime 55 To a solution of 5-trimethylsilanylethynyl-pyridin-2- ylamine (26 mg, 137 mmol) described in Manufacturing Example 28-1-2 in tetrahydrofuran (1 mL) and methanol (1 OH mL) was added potassium carbonate (37.9 mg, 274 mmol) at room temperature, which was stirred for 1 hour at room 60 temperature. The reaction Solution was partitioned into water N H and ethyl acetate at 0°C. The organic layer was washed with saturated aqueous sodium chloride, and dried over anhydrous 2 magnesium sulfate, and the solvent was concentrated under a N NH2 reduced pressure. The residue was purified by NH silica gel 65 column chromatography (heptane:ethyl acetate-1: 1) to To a solution of 2-amino-3-formylpyridine (1.00 g, 8.19 obtain the title compound (16 mg, 99%). mmol) in pyridine (20 mL) was added hydroxylamine hydro US 8,158,657 B2 137 138 chloride (854 mg, 12.3 mmol) at room temperature, which ammonium chloride Solution was added to the reaction solu was stirred for 1 hour and 40 minutes at room temperature. tion at room temperature, which was then extracted with ethyl The reaction solution was partitioned into water and ethyl acetate. The organic layer was washed with Saturated aqueous acetate. The organic layer was washed with water and Satu Sodium chloride and dried over anhydrous magnesium Sul rated aqueous sodium chloride, and dried over anhydrous fate, and the solvent was evaporated under a reduced pressure. magnesium Sulfate, and the solvent was evaporated under a The residue was purified by NH silica gel column chroma reduced pressure. The residue was purified by NH silica gel tography (heptane:ethyl acetate=30:1) to obtain the title com column chromatography (ethyl acetate:methanol-10:1) to pound (91.1 mg, 72%). obtain the title compound (951 mg. 85%). H-NMR Spectrum (CDC1) & (ppm): 0.18 (9H, s), 3.59 H-NMR Spectrum (DMSO-d) & (ppm): 6.60 (1H, dd, 10 (2H, s), 5.06 (2H, s), 6.92-6.95 (2H, m), 7.23-7.26 (2H, m), J=48, 7.3 Hz), 6.94 (2H, s), 7.55 (1H, m), 7.96 (1H, dd, J=1.7, 7.30-7.34 (1H, m), 7.36-7.40 (2H, m), 7.42-7.44 (2H, m). 4.8 Hz), 8.22 (1H, s), 11.2 (1H, s). Manufacturing Example 29-2-2 Manufacturing Example 29-1-2 15 1-Benzyloxy-4-prop-2-ynyl-benzene 2-Amino-5-chloro-N-hydroxypyridin-3-carboxyimidoyl chloride 2^ OH

C N C 25 2 N NH2 To a solution of (3-(4-benzyloxy-phenyl)-prop-1-ynyl)-tri methyl-silane (91.1 mg, 3.09 mmol) described in Manufac turing Example 29-2-1 in methanol (20 mL) was added potas To a solution of 2-amino-pyridine-3-carbaldehyde oxime 30 sium carbonate (854 mg., 6.18 mmol) at room temperature, (95.1 mg, 6.93 mmol) described in Manufacturing Example which was stirred for 4 hours and 10 minutes at room tem 29-1-1 in N,N-dimethylformamide (20 mL) was added perature. The reaction solution was partitioned into water and N-chlorosuccinimide (2.22 g, 16.6 mmol) at room tempera ethyl acetate at room temperature. The organic layer was ture, which was stirred at room temperature for 5 hours and 30 washed with Saturated aqueous sodium chloride and dried minutes. The reaction solution was partitioned into water and 35 over anhydrous magnesium Sulfate, and the solvent was ethyl acetate at room temperature. The organic layer was evaporated under a reduced pressure. The residue was puri washed with water and Saturated aqueous sodium chloride, fied by silica gel column chromatography (heptane:ethyl and dried over anhydrous magnesium Sulfate, and the solvent acetate=20:1) to obtain the title compound (618 mg, 90%). was concentrated under a reduced pressure. The residue was H-NMR Spectrum (CDC1) (ppm): 8 2.16 (1H, t, J–2.4 purified by silica gel column chromatography (heptane:ethyl 40 acetate=1:1) to obtain the title compound (249 mg, 17%). HZ), 3.54 (2H, d, J=2.4 Hz), 5.05 (2H, s), 6.91-6.94 (2H, m), H-NMR Spectrum (DMSO-d) & (ppm): 7.24 (2H, brs), 7.24-7.26 (2H, m), 7.29-7.43 (5H, m). 7.91-792 (1H, m), 8.06-8.07 (1H, m), 12.6 (1H, s). Manufacturing Example 29-2-3 Manufacturing Example 29-2-1 45 3-(5-(4-Benzyloxy-benzyl)-isoxazol-3-yl)-5-chloro (3-(4-Benzyloxy-phenyl)-prop-1-ynyl)-trimethyl pyridin-2-ylamine silane

50

e 55 C S o N N

2 To a solution of trimethylsilyl acetylene (851 uL, 6.02 N NH2 mmol) in tetrahydrofuran (20 mL) was added ethyl magne 60 sium bromide (3 M diethyl ether solution, 1.86 mL, 5.59 mmol) under nitrogen atmosphere at room temperature, To a solution of 2-amino-5-chloro-N-hydroxypyridin-3- which was stirred for 40 minutes at 65° C. The reaction carboxyimidoyl chloride (100 mg. 485 umol) described in Solution was cooled to room temperature, and copper (I) Manufacturing Example 29-1-2 in diethyl ether (2 mL) and bromide (308 mg, 2.16 mmol) and 4-benzyloxybenzyl chlo 65 tetrahydrofuran (1 mL) were added 1-benzyloxy-4-prop-2- ride (1.00 g, 4.30 mmol) were added to the reaction solution ynyl-benzene (113 mg, 509 umol) described in Manufactur and stirred for 8 hours and 45 minutes at 65° C. Saturated ing Example 29-2-2 and triethylamine (81 uL, 582 umol), US 8,158,657 B2 139 140 which was stirred for 4 hours and 5 minutes at room tempera Manufacturing Example 30-1-1 ture. The reaction solution was concentrated under a reduced pressure. The residue was purified by NH silica gel column 2-(4-Chloromethyl-benzyloxy)-pyridine chromatography (heptane:ethyl acetate=5:1) to obtain the title compound (59 mg, 31%). H-NMR Spectrum (DMSO-d) & (ppm): 4.11 (2H, s), 5.07 (2H, s), 6.97-6.99 (3H, m), 7.05 (2H, s), 7.24 (2H, d, C J=8.6 Hz), 7.29-7.32 (1H, m), 7.37 (2H, m), 7.42 (2H, m), 8.07 (1H, d. J=2.6 Hz), 8.11 (1H, d, J=2.6 Hz). 10 Example 30

3-(5-(4-(Pyridin-2-yloxymethyl)-benzyl-isoxazol-3- 15 A mixture of (4-(pyridin-2-yloxymethyl)-phenyl)metha yl)-pyridin-2-ylamine nol (540 mg, 2.51 mmol) described in Manufacturing Example 2-1-1, triphenylphosphine (856 mg, 3.27 mmol) and carbon tetrachloride (10.8 g. 10.2 mmol) was stirred under reflux for 2 hours and 10 minutes. The reaction solution was cooled to room temperature, and concentrated under a reduced pressure. The residue was purified by silica gel col umn chromatography (heptane:acetic acid-8:1) to obtain the title compound (300 mg, 51.1%). H-NMR Spectrum (DMSO-d) & (ppm): 4.76 (2H, s), 25 5.35 (2H, s), 6.86-6.90 (1H, m), 6.97-7.20 (1H, m), 7.44 (4H, s), 7.70-7.76 (1H, m), 8.15-8.18 (1H m). e Manufacturing Example 30-1-2 N s / 30 2-(4-Prop-2-ynyl-benzyloxy)-pyridine 2 N NH

35 2^ To a solution of 5-chloro-3-(5-(4-(pyridin-2-yloxym ethyl)-benzyl)-isoxazol-3-yl)-pyridin-2-ylamine (37 mg, 94 umol) described in Manufacturing Example 30-1-3 in N-me thyl-2-pyrrolidinone (2 mL) were added formic acid (5.3 uL. 40 0.14 mmol), N,N-diisopropylethylamine (49 uL, 0.28 mmol) To a solution of trimethylsilyl acetylene (496 uL, 3.51 and bis(tri-tert-butylphosphine)palladium (O) (9.6 mg, 19 mmol) in tetrahydrofuran (15 mL) was added ethyl magne sium bromide (3 M diethyl ether solution, 1.09 mL, 3.28 umol) at room temperature, which was stirred under nitrogen mmol) under nitrogen atmosphere at room temperature, atmosphere for 1 hour and 25 minutes at 100° C. Water and 45 which was stirred for 30 minutes at 65° C. The reaction ethyl acetate were added to the reaction solution at room Solution was cooled to room temperature, and copper (I) temperature, which was then filtered through a Celite pad. bromide (168 mg, 1.17 mmol) and 2-(4-chloromethyl-benzy The filtrate was partitioned into water and ethyl acetate. The loxy)-pyridine (548 mg, 2.34 mmol) manufactured in Manu organic layer was washed with water and Saturated aqueous facturing Example 30-1-1 were added thereto and stirred for Sodium chloride, and dried over anhydrous magnesium Sul 50 15 hours and 10 minutes at 65° C. The reaction solution was fate, and the solvent was evaporated under a reduced pressure. partitioned into Saturated aqueous ammonium chloride solu The residue was purified by reverse-phase high performance tion and ethyl acetate at room temperature. The organic layer liquid chromatography (using an acetonitrile-water mobile was washed with Saturated aqueous sodium chloride, and phase containing 0.1% trifluoroacetic acid), and then further dried over anhydrous magnesium sulfate, and the solvent was purified by silica gel column chromatography (heptane:ethyl 55 evaporated under a reduced pressure. To a solution of the acetate=1:1) to obtain the title compound (0.66 mg, 2.0%). resulting residue in methanol (5 mL) and tetrahydrofuran (10 H-NMR Spectrum (CDC1) & (ppm): 4.14 (2H, s), 5.39 mL) was added potassium carbonate (647 mg, 4.68 mmol), (2H, s), 6.27 (1H, s), 6.49 (2H, brs), 6.69 (1H, dd, J–4.9, 7.5 which was stirred for 3 hours and 25 minutes at room tem Hz), 6.81 (1H, d, J=8.4 Hz), 6.88-6.91 (1H, m), 7.31 (2H, d, perature. The reaction Solution was partitioned into water and 60 ethyl acetate at room temperature. The organic layer was J=8.0 Hz), 7.47 (2H, d, J=8.0 Hz), 7.57-7.62 (1H, m), 7.68 washed with Saturated aqueous sodium chloride and dried (1H, dd, J=1.8, 7.5 Hz), 8.09 (1H, dd, J=1.8, 4.9 Hz), 8.17 over anhydrous magnesium Sulfate, and the solvent was 8.19 (1H, m). evaporated under a reduced pressure. The residue was puri MS m/e (ESI) 359.11 (MH) fied by silica gel column chromatography (heptane:ethyl The starting material, 5-chloro-3-(5-(4-(pyridin-2- 65 acetate=20:1) to obtain a mixture of the title compound and yloxymethyl)-benzyl)-isoxazol-3-yl)-pyridin-2-ylamine, 2-(4-chloromethyl-benzyloxy)-pyridine (448 mg. target was synthesized as follows. purity 20%, 17%). US 8,158,657 B2 141 142 H-NMR Spectrum (DMSO-d) & (ppm): 3.04 (1H, m), nitrogen atmosphere, which was stirred for 4 hours at 70° C. 3.61 (2H, d, J–2.6 Hz), 5.30 (2H, s), 6.83-6.87 (1H, m), Water and ethyl acetate were added to the reaction mixture at 6.95-6.99 (1H, m), 7.30-7.32 (2H, s), 7.36-7.40 (2H, m), room temperature, which was then filtered through a Celite 7.68-7.73 (1H, m), 8.14-8.16 (1H, m). pad, and the filtrate was partitioned into water and ethyl acetate. The organic layer was washed with water and Satu Manufacturing Example 30-1-3 rated aqueous Sodium chloride, and dried over anhydrous magnesium sulfate, and the solvent was evaporated under a 5-Chloro-3-(5-(4-(pyridin-2-yloxymethyl)-benzyl) reduced pressure. The residue was purified by NH silica gel isoxazol-3-yl)-pyridin-2-ylamine column chromatography (ethyl acetate:heptane 2:1, then 10 ethyl acetate) to obtain the title compound (1.8 mg, 2%). 'H-NMR Spectrum (DMSO-d) & (ppm): 5.09 (2H, s), 5.26 (2H, s), 5.87 (2H, brs), 6.61 (1H, dd, J–4.8, 7.2 Hz), 6.98 Na (2H, d, J=8.8 Hz), 7.27 (2H, d, J=8.8 Hz), 7.32-7.44 (5H, m), 15 747-749 (1H, m), 7.74 (1H, s), 7.86 (1H, dd, J=1.6, 5.0 Hz), o-N h 8.13 (1H, s). The starting material, 1-(4-benzyloxy-benzyl)-4-tributyl e Stannanyl-1H-pyrazole, was synthesized as follows. Manufacturing Method 31-1-1 1-(4-Benzyloxy-benzyl)-4- C N1SN bromo-1H-pyrazole

2 N NH2

To a solution of 2-amino-5-chloro-N-hydroxypyridine-3- 25 carboxyimidoyl chloride (50 mg, 242 umol) described in Manufacturing Example 29-1-2 in tetrahydrofuran (5 mL) N were added triethylamine (41 uL. 292 umol) and 2-(4-prop 2-ynyl-benzyloxy)-pyridine (271 mg, 243 Limol, purity: 20%) M described in Manufacturing Example 30-1-2 at room tem 30 Br O perature, which was stirred for 30 minutes at room tempera ture, and further stirred under reflux for 2 hours and 25 min To a solution of 2-bromopyrazole (500 mg, 3.40 mmol) in utes. The reaction solution was cooled to room temperature, N,N-dimethylformamide (10 mL) was added sodium hydride and concentrated under a reduced pressure. The residue was (196 mg, 4.08 mmol. 60% in oil) on an ice bath (0°C.) under purified by NH silica gel column chromatography (heptane: 35 nitrogen atmosphere, which was stirred for 30 minutes at ethyl acetate=5:1) to obtain the title compound (37 mg,39%). room temperature. 4-Benzyloxybenzyl chloride (791 mg, H-NMR Spectrum (DMSO-d) & (ppm): 4.22 (2H, s), 3.40 mmol) was then added and stirred for 60 minutes at room 5.34 (2H, s), 6.86 (1H, d. J=8.2 Hz), 6.97-7.01 (1H, m), 7.04 temperature. The reaction mixture was partitioned into water (1H, s), 7.07 (2H, brs), 7.34 (2H, d, J=8.0 Hz), 7.44 (2H, d, 40 and ethyl acetate at room temperature. The organic layer was J=8.0 Hz), 7.70-7.74 (1H, m), 8.09 (1H, d, J=2.6 Hz), 8.14 washed with water and Saturated aqueous sodium chloride, (1H, d, J=2.6 Hz), 8.16-8.18 (1H, m). and dried over anhydrous magnesium Sulfate, and the solvent Example 31 was evaporated under a reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate: 3-(1-(4-Benzyloxy-benzyl)-1H-pyrazol-4-yl)-pyri 45 heptane=1:5) to obtain the title compound (1.1 g, 94%). din-2-ylamine 'H-NMR Spectrum (DMSO-d) & (ppm): 5.04 (2H, s), 5.17 (2H, s), 6.94 (2H, d, J=8.8 Hz), 7.17 (2H, d, J=8.8 Hz), 7.31 (1H, s), 7.33-741 (5H, m), 7.47 (1H, m). Manufacturing Example 31-1-2 1-(4-Benzyloxy-benzyl)-4-tributylstannanyl-1H pyrazole

55

60 To a solution of 2-amino-3-bromopyridine (44.1 mg 0.26 mmol) in anhydrous tetrahydrofuran (7 mL) were added 1-(4- benzyloxy-benzyl)-4-tributylstannanyl-1H-pyrazole (141 mg, 0.26 mmol) described in Manufacturing Example 31-1- 65 2, copper (I) iodide (19.4 mg., 0.10 mmol) and bis(triph enylphosphine)palladium (II) (35.8 mg, 0.05 mmol) under US 8,158,657 B2 143 144 To a solution of 1-(4-benzyloxy-benzyl)-4-bromo-1H The starting material, 3-(1H-pyrazol-4-yl)-pyridin-2- pyrazole (1.10 g, 3.20 mmol) described in Manufacturing ylamine, was synthesized as follows. Example 31-1-1 in Xylene (20 mL) were added tetrakis(triph enylphosphine)palladium (O) (370 mg, 0.32 mmol) and hexa Manufacturing Example 32-1-1 n-butyl stannane (5.57 g., 9.60 mmol) under nitrogen atmo sphere, which was stirred for 2 hours at 140° C. Water and 4-Bromo-1-trityl-1H-pyrazole ethyl acetate were added to the reaction mixture at room temperature, which was then filtered through a Celitepad, and the filtrate was partitioned into water and ethyl acetate. The organic layer was washed with water and Saturated aqueous 10 Sodium chloride, and dried over anhydrous magnesium Sul fate, and the solvent was evaporated under a reduced pressure. The residue was purified by silica gel column chromatogra phy (ethyl acetate:heptane=1:5) to obtain the title compound 15 (141 mg, 8%). N H-NMR Spectrum (CDC1) & (ppm): 0.87 (9H, t, J–7.2 Hz), 0.92-1.00 (6H, m), 1.26-1.35 (6H, m), 1.4.6-1.54 (6H, m), 5.05 (2H, s), 5.27 (2H, s), 6.93-6.95 (2H, m), 7.14-7.17 (2H, m), 7.23 (1H, s), 7.31-743 (5H, m), 7.46 (1H, s). B To a solution of 4-bromopyrazole (10.0 g, 68.0 mmol) in Example 32 N,N-dimethylformamide (100 mL) was added dropwise tri ethylamine (23.7 mL, 170 mmol) under nitrogen atmosphere 3-(1-(4-(Pyridin-2-yloxymethyl)-benzyl)-1H-pyra 25 at room temperature. Trityl chloride (37.9 g, 136 mmol) was Zol-4-yl)-pyridin-2-ylamine added to the reaction solution on an ice bath (0° C.), and stirred for 3 hours at 70° C. Water (400 mL) was added to the reaction Solution to precipitate the Solids. The precipitated solids were filtered and dried under a reduced pressure. The 30 solids were then azeotropically dried with toluene to obtain e the title compound (22.9 g, 87%). o-Q H-NMR Spectrum (DMSO-d) & (ppm): 7.04-7.07 (6H, N m), 7.35-7.38 (9H, m), 7.52 (1H, d, J=0.4 Hz), 7.76 (1H, d, N J=0.8 Hz). 35 X Manufacturing Example 32-1-2 N 4-(4.4.5,5-Tetramethyl-(1,3,2)dioxaborolan-2-yl)-1- trity1-1H-pyrazole N NH2 40

To a solution of 3-(1H-pyrazol-4-yl)-pyridin-2-ylamine (150 mg. 0.94 mmol) described in Manufacturing Example 45 32-1-4 in N,N-dimethylformamide (10 mL) was added sodium hydride (48.7 mg, 1.22 mmol. 60% in oil) on an ice bath (0°C.) under nitrogen atmosphere. Following 40 min utes of stirring at room temperature, 2-(4-chloromethyl-ben Zyloxy)-pyridine (228 mg, 0.98 mmol) described in Manu 50 facturing Example 30-1-1 was added and stirred for 30 minutes at room temperature. The reaction mixture was par titioned into water and ethyl acetate at room temperature. The organic layer was washed with water and Saturated aqueous Sodium chloride, and dried over anhydrous magnesium Sul 55 fate, and the solvent was evaporated under a reduced pressure. The residue was purified by NH silica gel column chroma tography (ethyl acetate: heptane-2:1, then ethyl acetate) to A mixture of 4-bromo-4-trity1-1H-pyrazole (4.8 g. 12.3 obtain the title compound (307 mg, 92%). 60 mmol) described in Manufacturing Example 32-1-1, bis(pi 'H-NMR Spectrum (DMSO-d) & (ppm): 5.33 (2H, s), nacolate)diboran (5.0 g, 19.7 mmol), potassium acetate (3.62 5.35 (2H, s), 5.60 (2H, brs), 6.61 (1H, dd, J–4.8, 7.4 Hz), g,36.9 mmol.), 1,1'bis(diphenylphosphino) ferrocene dichlo 6.84-6.87 (1H, m), 6.96-7.00 (1H, m), 7.30-7.43 (1H, m), ropalladium(II) (450 mg, 0.62 mmol) and dimethylsulfoxide 7.31 (2H, d. J=8.4 Hz), 7.42 (1H, t, J=8.4 Hz), 7.48 (1H, dd, (50 mL) was stirred under argon atmosphere for 17 hours and J=2.0, 7.2 Hz), 7.69-7.73 (1H, m), 7.76 (1H, d, J=1.2 Hz), 65 10 minutes at 80° C. The reaction solution was allowed to 7.87 (1H, dd, J-2.0, 5.0 Hz), 8.15-8.17 (1H, m), 8.18 (1H, d, room temperature, and partitioned into water and ethyl J=0.8 Hz). acetate. The organic layer was concentrated under a reduced US 8,158,657 B2 145 146 pressure. The residue was purified by silica gel chromatog under a reduced pressure. The residue was purified by silica raphy (heptane:ethyl acetate-4:1). Heptane was added to the gel chromatography (ethyl acetate, then ethyl acetate:metha Solids obtained by concentrating the eluate under a reduced nol=10:1) to obtain the title compound (625 mg, 68.3%). pressure, which were then irradiated by ultrasonic wave and H-NMR Spectrum (DMSO-d) & (ppm): 5.59 (2H, brs), filtered to obtain the title compound (1.51 g, 28.0%). 5 6.62(1H, dd, J=4.8, 7.6Hz), 7.49 (1H, d, J=7.2Hz), 7.88 (1H, H-NMR Spectrum (CDC1) & (ppm): 1.30 (12H, s), 7.10 d, J=4.8 Hz), 7.72-8.15 (2H, brs), 12.9 (1H, brs). 7.16 (6H, m), 7.26-7.31 (9H, m), 7.75 (1H, s), 7.94 (1H, s). Example 33 Manufacturing Example 32-1-3 10 3-(1-(4-Butoxymethyl-benzyl)-1H-pyrazol-4-yl)- 3-(1-Trity1-1H-pyrazol-4-yl)-pyridin-2-ylamine pyridin-2-ylamine

15 N

25 To a mixture of 3-(1H-pyrazol-4-yl)-pyridin-2-ylamine 4-(4.4.5.5-Tetramethyl-(1,3,2)dioxaborolan-2-yl)-1-tri (20 mg, 0.13 mmol) described in Manufacturing Example tyl-1H-pyrazole (3.2g, 7.33 mmol) described in Manufactur 32-1-4 and N,N-dimethylformamide (1 mL) was added ing Example 32-1-2,3-bromo-pyridine-2-ylamine (1.14 g. 30 sodium hydride (6.8 mg, 0.19 mmol. 66% in oil), which was 6.60 mmol), tetrakis(triphenylphosphine)palladium (O) (424 stirred for 30 minutes at room temperature. 1-Butoxymethyl mg, 0.37 mmol), toluene (40 mL), 2 M aqueous sodium 4-chloromethyl-benzene (29 mg 0.14 mmol) described in carbonate solution (10 mL) and ethanol (20 mL) were stirred Manufacturing Example 33-1-4 was added to the reaction for 1 hour at 95°C. The reaction solution was allowed to room mixture at room temperature, and stirred for 1.5 hours at 40° temperature, and partitioned into water and ethyl acetate. The 35 C. The reaction mixture was cooled and partitioned into water ethyl acetate layer was washed with water once, and the and ethyl acetate. The organic layer was washed with Satu Solvent was evaporated under a reduced pressure. The residue rated aqueous Sodium chloride, and the solvent was evapo was purified by Silica gel chromatography (heptane:ethyl rated under a reduced pressure. The residue was purified by acetate=1:2) to obtain the title compound (2.3 g 78.0%). NH silica gel column chromatography (ethyl acetate:hep H-NMR Spectrum (DMSO-ds) & (ppm): 5.52 (2H, brs), 40 tane=2:1) to obtain the title compound (33 mg, 78%). 6.57 (1H, dd, J=7.2, 4.8 Hz), 7.10-7.16 (6H, m), 7.28-7.38 H-NMR Spectrum (CDC1) & (ppm): 0.92 (3H, t, J–7.4 (9H, m), 7.42 (1H, d, J=7.2 Hz), 7.66 (1H, s), 7.84 (1H, d, Hz), 1.35-1.44 (2H, m), 1.56-1.62 (2H, m), 3.48 (2H, t, J–6.6 J–4.8 Hz), 7.92 (1H, s). Hz), 4.49 (2H, s), 4.61 (2H, brs), 5.34 (2H, s), 6.70 (1H, dd, J=5.0, 7.4 Hz), 7.27 (2H, d, J=8.1 Hz), 7.35 (2H, d, J=8.1 Hz), Manufacturing Example 32-1-4 45 7.39 (1H, dd, J=1.8, 7.3 Hz), 7.58 (1H, s), 7.73 (1H, d, J=0.7 Hz), 8.00 (1H, dd, J=1.8, 5.1 Hz). 3-(1H-Pyrazol-4-yl)-pyridin-2-ylamine The starting material, 1-butoxymethyl-4-chloromethyl benzene, was synthesized as follows.

50 Manufacturing Example 33-1-1 X 4-Butoxymethyl-benzonitrile 55 s2 N NH2

3-(1-Trityl-1H-pyrazol-4-yl)-pyridin-2-ylamine (2.3 g, 5.71 mmol) described in Manufacturing Example 32-1-3, 2N 60 N-1S-1 hydrochloric acid (15 mL), methanol (15 mL) and tetrahy drofuran (10 mL) were stirred for 30 minutes at 70° C. The To a mixture of sodium hydride (270 mg, 11 mmol. 66% in reaction Solution was allowed to room temperature, and par oil) and tetrahydrofuran (20 mL) was added n-butanol (1.1 titioned into water and ethyl acetate. Saturated sodium bicar mL, 12 mmol) at 0°C., which was stirred for 45 minutes at bonate Solution was added to the separated aqueous layer, 65 room temperature. The reaction mixture was cooled to 0°C., which was then extracted with ethyl acetate 6 times. The ethyl and a mixture of 4-cyanobenzyl bromide (1.5 g, 7.4 mmol) acetate layers were combined and the solvent was evaporated and tetrahydrofuran (10 mL) was added dropwise at that US 8,158,657 B2 147 148 temperature. The reaction mixture was stirred for 3 hours at H-NMR Spectrum (CDC1) & (ppm): 0.92 (3H, t, J–7.3 room temperature, and N,N-dimethylformamide (10 mL) Hz), 1.35-1.44 (2H, m), 1.57-1.64 (2H, m), 3.47 (2H, t, J–6.6 was added to the reaction mixture and stirred for further 4.5 Hz), 4.50 (2H, s), 4.69 (2H, s), 7.34 (4H, s). hours at the same temperature. The reaction mixture was partitioned into water and diethyl ether. The organic layer was Manufacturing Example 33-1-4 washed with Saturated aqueous sodium chloride, and the Sol vent was evaporated under a reduced pressure. The residue 1-Butoxymethyl-4-chloromethyl-benzene was purified by NH silica gel column chromatography (ethyl acetate:heptane-1:6) to obtain the title compound (1.2 g, 84%). 10 'H-NMR Spectrum (CDC1) & (ppm): 0.93 (3H, t, J–7.3 C Hz), 1.37-1.46 (2H, m), 1.59-1.66 (2H, m), 3.50 (2H, t, J–6.6 Hz), 4.55 (2H, s), 743-746 (2H, m), 7.62-7.65 (2H, m). N-1 nu-1 Manufacturing Example 33-1-2 15 A mixture of (4-butoxymethyl-phenyl)-methanol (190 mg. 4-Butoxymethyl-benzylamine 0.98 mmol) described in Manufacturing Example 33-1-3, triphenylphosphine (310 mg, 1.2 mmol) and carbon tetra chloride (3 mL) was stirred under reflux for 7 hours. The reaction mixture was cooled to room temperature, and con centrated under a reduced pressure. The residue was purified HN by neutral silica gel column chromatography (ethyl acetate: heptane=1:15) to obtain the title compound (180 mg, 86%). O n1N1 25 H-NMR Spectrum (CDC1) & (ppm): 0.92 (3H, t, J–7.3 Hz), 1.35-1.45 (2H, m), 1.57-1.64 (2H, m), 3.47 (2H, t, J–6.6 To a mixture of lithium aluminum hydride (600 mg, 13 Hz), 4.50 (2H, s), 4.59 (2H, s), 7.32-7.38 (4H, m). mmol, purity: 80%) and tetrahydrofuran (10 mL) was added a mixture of 4-butoxymethyl-benzonitrile (600 mg, 3.2 Example 34 mmol) described in Manufacturing Example 33-1-1 and tet 30 rahydrofuran (10 mL) at 0°C., which was stirred for 4 hours 3-(1-(4-Phenoxy-benzyl)-1H-pyrazol-4-yl)-pyridin at room temperature. 28% Aqueous ammonia solution was 2-ylamine added dropwise to the reaction mixture at OD. The reaction mixture was warmed to room temperature and filtered. The filtrate was concentrated under a reduced pressure to obtain 35 the title compound (620 mg, 10.1%) as a crude product. O H-NMR Spectrum (CDC1) & (ppm): 0.92 (3H, t, J–7.3 Hz), 1.37-1.44 (2H, m), 1.56-1.63 (2H, m), 3.47 (2H, t, J–6.6 HZ), 3.86 (2H, s), 4.49 (2H, s), 7.27-7.32 (4H, m). N 40 Manufacturing Example 33-1-3 MN

(4-Butoxymethyl-phenyl)-methanol r2 N NH2 45 To a solution of 3-(1H-pyrazol-4-yl)-pyridin-2-ylamine HO (20 mg, 0.13 mmol) described in Manufacturing Example 32-1-4 in N,N-dimethylformamide (10 mL) was added N-1-1 50 sodium hydride (7.5 mg 0.19 mmol. 60% in oil) under nitro gen atmosphere on an ice bath (0°C.), which was stirred for 30 minutes at room temperature. 1-Chloromethyl-4-phe To a mixture of 4-butoxymethyl-benzylamine (250 mg, 1.3 noxy-benzene (32.8 mg, 0.15 mmol) described in Manufac mmol) described in Manufacturing Example 33-1-2, acetic turing Example 34-1-1 was then added to the mixture and acid (2 mL) and water (2 mL) was added sodium nitrite (1.1 55 stirred for 30 minutes at room temperature. The reaction g, 16 mmol) at 0°C., which was stirred for 40 minutes at room mixture was partitioned into water and ethyl acetate at room temperature. The reaction mixture was partitioned into ethyl temperature. The organic layer was washed with water and acetate and water. The organic layer was washed with Satu saturated aqueous Sodium chloride, and dried over anhydrous rated aqueous Sodium hydrogen carbonate Solution and Satu magnesium sulfate, and the solvent was evaporated under a rated aqueous Sodium chloride, and the solvent was evapo 60 reduced pressure. The residue was purified by NH silica gel rated under a reduced pressure. Methanol (2 mL) and column chromatography (ethyl acetate:heptane 2:1, then potassium carbonate (360 mg, 2.6 mmol) were added to the ethyl acetate only) to obtain the title compound (41 mg, 86%, residue, and the reaction mixture was stirred for 1.5 hours at purity: 90%). room temperature. The reaction mixture was concentrated H-NMR Spectrum (DMSO-d) & (ppm): 5.33 (2H, s), under a reduced pressure. The residue was purified by neutral 65 5.60 (2H, brs), 6.61 (1H, dd, J=4.8, 7.4 Hz), 6.98-7.01 (2H, silica gel column chromatography (ethyl acetate: heptane-1: m), 7.12-7.16 (1H, m), 7.34-7.40 (2H, m), 7.48-7.65 (5H, m), 1) to obtain the title compound (200 mg, 78%). 7.77 (1H, s), 7.87 (1H, dd, J=1.2, 5.0 Hz), 8.18 (1H, s). US 8,158,657 B2 149 150 The starting material, 1-chloromethyl-4-phenoxy-ben The starting material, 1-chloromethyl-3-phenoxy-ben Zene, was synthesized as follows. Zene, was synthesized as follows. Manufacturing Example 34-1-1 Manufacturing Example 35-1-1 1-Chloromethyl-4-phenoxy-benzene 1-Chloromethyl-3-phenoxy-benzene

10 C C

O

To a solution of (4-phenoxy-phenyl)-methanol (408 mg, 15 To a solution of (3-phenoxy-phenyl)-methanol (2.00 g, 2.04 mmol) in carbon tetrachloride (8.2 mL) was added triph 10.0 mmol) in carbon tetrachloride (40 mL) was added triph enylphosphine (642 mg, 2.45 mmol) under nitrogen atmo enylphosphine (3.15g, 12.0 mmol) at room temperature. The sphere at room temperature, and the reaction solution was reaction Solution was stirred under nitrogen atmosphere for 5 stirred under reflux for 7 hours and 40 minutes. The reaction hours and 40 minutes under reflux. The reaction mixture was mixture was cooled to room temperature and concentrated cooled to room temperature and concentrated under a reduced under a reduced pressure. The residue was purified by silica pressure. The residue was purified by silica gel column chro gel column chromatography (heptane:ethyl acetate 10:1) to matography (heptane:ethyl acetate=10:1) to obtain the title obtain the title compound (409 mg, 92%). compound (2.05 g, 94%). 'H-NMR Spectrum (DMSO-d) & (ppm): 4.76 (2H, s), 25 'H-NMR Spectrum (DMSO-d) & (ppm): 4.37 (2H, s), 6.98-7.05 (4H, m), 7.15-7.19 (1H, m), 7.39-7.46 (4H, m). 6.94-6.97 (1H, m), 7.00-7.03 (2H, m), 705-7.06 (1H, m), 7.13-7.20 (3H, m), 7.37-7.41 (2H, m). Example 35 Example 36 3-(1-(3-Phenoxy-benzyl)-1H-pyrazol-4-yl)-pyridin 30 2-ylamine 3-(1-(4-Benzyloxy-benzyl)-1H-pyrazol-4-yl)-pyri din-2,6-diamine

35

N O MN 40

r2 N NH2 45 HN N To a solution of 3-(1H-pyrazol-4-yl)-pyridin-2-ylamine (20 mg, 0.13 mmol) described in Manufacturing Example To a solution of 3-(1H-pyrazol-4-yl)-pyridin-2,6-diamine 32-1-4 in N,N-dimethylformamide (10 mL) was added (25 mg, 0.14 mmol) described in Manufacturing Example sodium hydride (7.5 mg, 0.19 mmol. 60% in oil) under nitro 50 36-1-2 in N,N-dimethylformamide (10 mL) was added gen atmosphere on an ice bath (0°C.), which was stirred for sodium hydride (8.6 mg, 0.22 mmol. 60% in oil) under nitro 40 minutes at room temperature. 1-Chloromethyl-3-phe gen atmosphere on an ice bath (0°C.), which was stirred for noxybenzene (32.8 mg, 0.15 mmol) described in Manufac 30 minutes at room temperature. 4-Benzyloxybenzyl chlo turing Example 35-1-1 was then added and stirred for 30 ride (49.9 mg, 0.22 mmol) was then added and stirred for 30 minutes at room temperature. The reaction mixture was par 55 minutes at room temperature. The reaction mixture was par titioned into water and ethyl acetate at room temperature. The titioned into water and ethyl acetate at room temperature. The organic layer was washed with water and Saturated aqueous organic layer was washed with water and Saturated aqueous Sodium chloride, and dried over anhydrous magnesium Sul Sodium chloride, and dried over anhydrous magnesium Sul fate, and the solvent was evaporated under a reduced pressure. fate, and the solvent was evaporated under a reduced pressure. The residue was purified by NH silica gel column chroma 60 The residue was purified by NH silica gel column chroma tography (ethyl acetate:heptane 2:1, then ethyl acetate only) tography (ethyl acetate:heptane 2:1, then ethyl acetate only) to obtain the title compound (20 mg, 47%). to obtain the title compound (24.0 mg. 45%). 'H-NMR Spectrum (DMSO-d) & (ppm): 5.35 (2H, s), 'H-NMR Spectrum (DMSO-d) & (ppm): 5.06 (2H, brs), 5.59 (2H, brs), 6.62 (1H, dd, J=1.2, 7.4 Hz), 6.90-6.95 (2H, 5.09 (2H, s), 5.21 (2H, s), 5.43 (2H, brs), 5.77 (1H, d, J=8.0 m), 6.99-7.06(3H, m), 7.13-7.17 (1H, m), 7.34-7.41 (3H, m), 65 Hz), 6.97-7.00 (2H, m), 7.15 (1H, d, J=8.0 Hz), 7.23-7.26 7.48 (1H, dd, J=2.0, 7.4 Hz), 7.70 (1H, d, J=0.8 Hz), 7.87 (1H, (2H, m), 7.30-7.34 (1H, m), 7.36-7.44 (4H, m), 7.56 (1H, d, dd, J=2.0, 5.0 Hz), 8.18 (1H, d, J=0.8 Hz). J=1.2 Hz), 7.90 (1H, d, J=1.2 Hz). US 8,158,657 B2 151 152 The starting material, 3-(1H-pyrazol-4-yl)-pyridin-2,6-di due was purified by NH silica gel column chromatography amine, was synthesized as follows. (ethyl acetate, then ethyl acetate:methanol=10:1) to obtain the title compound (600 mg, 60%). Manufacturing Example 36-1-1 'H-NMR Spectrum (DMSO-d) & (ppm): 5.04 (2H, brs), 5.41 (2H, brs), 5.78 (1H, d, J=8.4 3-(1-Trity1-1H-pyrazol-4-yl)-pyridin-2,6-diamine Hz), 7.16 (1H, d, J=8.0 Hz), 7.62 (1H, brs), 7.78 (1H, brs), 12.8 (1H, brs). Example 37 10 3-(1-(4-(Pyridin-2-yloxymethyl)-benzyl)-1H-pyra Zol-4-yl)-pyridin-2,6-diamine

15 e O-N f N NX HN r 2 HN N NH2 To a solution of 3-iodo-pyridin-2,6-diamine (3.3 g, 7.74 25 mmol, purity: 70%) described in Manufacturing Example To a solution of 3-(1H-pyrazol-4-yl)-pyridin-2,6-diamine 13-1-1 in toluene (50 mL) were added ethanol (25 mL), 2 N aqueous sodium carbonate Solution (12.5 mL), 4-(4.4.5.5- (25 mg, 0.14 mmol) described in Manufacturing Example tetramethyl-(1,3,2)dioxaborolan-2-yl)-1-trity1-1H-pyrazole 36-1-2 in N,N-dimethylformamide (3 mL) was added sodium 30 hydride (8.6 mg, 0.22 mmol. 60% in oil) under nitrogen (3.3 g, 7.56 mmol) described in Manufacturing Example atmosphere on an ice bath (0°C.). Following 30 minutes of 32-1-2 and tetrakis(triphenylphosphine)palladium (0) (1.02 stirring at room temperature, 2-(4-chloromethyl-benzyloxy)- g, 0.88 mmol) under nitrogen atmosphere, which was stirred pyridine (43.4 mg., 0.19 mmol) described in Manufacturing for 2.5 hours at 95°C. Water and ethyl acetate were added to Example 30-1-1 was added and stirred for 30 minutes at 60° the reaction mixture at room temperature, which was then 35 C. The reaction mixture was partitioned into water and ethyl filtered through a Celite pad, and the filtrate was partitioned acetate at room temperature. The organic layer was washed into water and ethyl acetate. The organic layer was washed with water and saturated aqueous sodium chloride, and dried with water and saturated aqueous sodium chloride, and dried over anhydrous magnesium Sulfate, and the solvent was over anhydrous magnesium Sulfate, and the solvent was evaporated under a reduced pressure. The residue was puri evaporated under a reduced pressure. The residue was puri 40 fied by NH silica gel column chromatography (ethyl acetate: fied by NH silica gel column chromatography (ethyl acetate: heptane=2:1, then ethyl acetate only) to obtain the title com heptane=1:2 then 2:1 then 5:1) to obtain the title compound pound (22.8 mg, 43%). (2.4g, 73%). 'H-NMR Spectrum (DMSO-d) & (ppm): 5.07 (2H, brs), 'H-NMR Spectrum (CDC1) & (ppm): 4.63 (2H, brs), 4.79 5.30 (2H, s), 5.32 (2H, s), 5.43 (2H, brs), 5.78 (1H, d, J=8.0 (2H, brs), 5.90 (1H, d, J=8.0 Hz), 7.16-7.20 (6H, m), 7.29 45 Hz), 6.84-6.86 (1H, m), 6.96-7.00 (1H, m), 7.16 (1H, d, J-8.0 7.32 (10H, m), 7.45 (1H, s), 7.77 (1H, s). Hz), 7.28 (2H, d, J=8.0 Hz), 7.41 (2H, d, J=7.6 Hz), 7.58 (1H, Manufacturing Example 36-1-2 s), 7.69-7.73 (1H, m), 7.94 (1H, s), 8.15-8.17 (1H, m). Example 38 3-(1H-pyrazol-4-yl)-pyridin-2,6-diamine 50 3-(1-(4-Butoxymethyl-benzyl)-1H-pyrazol-4-yl)- pyridin-2,6-diamine

55 N HN N 60 N To a solution of 3-(1-trity1-1H-pyrazol-4-yl)-pyridin-2,6- MN diamine (10.0 g, 25.7 mmol) described in Manufacturing Example 36-1-1 in methylene chloride (14 mL) was added r 2 trifluoroacetic acid (7 mL) under nitrogen atmosphere, which 65 HN N NH2 was stirred for 1 hour at room temperature. The reaction mixture was concentrated under a reduced pressure. The resi US 8,158,657 B2 153 154 To a solution of 3-(1H-pyrazol-4-yl)-pyridin-2,6-diamine The starting material, 3-(4-bromo-pyrazol-1-yl)-pyridin (20 mg, 0.11 mmol) described in Manufacturing Example 2-ylamine, was synthesized as follows. 36-1-2 in N,N-dimethylformamide (4 mL) was added sodium hydride (5.9 mg, 0.15 mmol. 60% in oil) under nitrogen Manufacturing Example 39-1-1 atmosphere on an ice bath (0°C.) Following 30 minutes of 5 stirring at room temperature, 1-butoxymethyl-4-chlorom 2,2-Dimethyl-N-pyridin-2-yl-propionamide ethyl-benzene (26.7 mg, 0.13 mmol) described in Manufac turing Example 33-1-4 was added and stirred for 30 minutes at room temperature. The reaction mixture was partitioned into water and ethyl acetate at room temperature. The organic 10 layer was washed with water and Saturated aqueous sodium chloride, and dried over anhydrous magnesium sulfate, and the solvent was evaporated under a reduced pressure. The residue was purified by NH silica gel column chromatogra phy (ethyl acetate:heptane=2:1, then ethyl acetate) to obtain 15 the title compound (29.0 mg, 72%). H-NMR Spectrum (DMSO-d) & (ppm): 0.864 (3H, d, J=7.6Hz), 1.30-1.35 (2H, m), 1.47-1.54 (2H, m), 3.40 (2H, d, To a solution of 2-aminopyridine (50.0 g, 531 mmol) in J=6.4 Hz), 4.42 (2H, s), 5.07 (2H, brs), 5.29 (2H, s), 5.43 (2H, methylene chloride (500 mL) were added triethylamine (81.4 brs), 5.78 (1H, d, J=8.4 Hz), 7.16 (1H, t, J=8.0Hz), 7.24-7.29 mL, 584 mmol) and pivaloyl chloride (71.9 mL, 584 mmol) at 0° C., which was stirred for 4 hours and 30 minutes at room (4H, m), 7.58 (1H, s), 7.93 (1H, s). temperature. The reaction Solution was partitioned into water and methylene chloride. The organic layer was washed with Example 39 25 water and Saturated aqueous sodium chloride, and dried over anhydrous magnesium Sulfate, and the solvent was evapo rated under a reduced pressure. To a solution of the resulting 3-(4-(4-Benzyloxy-benzyl)-pyrazol-1-yl)-pyridin-2- residue in methanol (300 mL) was added potassium carbon ylamine ate (73.4 g. 531 mmol) at 0° C., which was stirred for 90 30 minutes at room temperature. The reaction Solution was par titioned into water and ethyl acetate at room temperature. The organic layer was washed with saturated aqueous sodium chloride and dried over anhydrous magnesium Sulfate, and the solvent was evaporated under a reduced pressure. Heptane 35 (300 mL) was added to the residue, and the precipitated solids were filtered to obtain the title compound (80.2g, 85%). The filtrate was then concentrated under a reduced pressure, and the residue was purified by Silica gel column chromatography (heptane:ethyl acetate=2:1) to obtain the title compound 40 (12.2g, 13%). H-NMR Spectrum (DMSO-d) & (ppm): 1.22 (9H, s), 7.06-7.09 (1H, m), 7.72-7.77 (1H, m), 8.01-8.03 (1H, m), 8.29-8.31 (1H, m), 9.71 (1H, s).

45 Manufacturing Example 39-1-2 N-(3-Iodo-pyridin-2-yl)-2,2-dimethyl-propionamide

To a mixture of 3-(4-bromo-pyrazol-1-yl)-pyridin-2- ylamine (34 mg., 0.14 mmol) described in Manufacturing 50 Example 39-1-4 (4-benzyloxy-benzyl)-tributyl-stannane (84 mg., 0.17 mmol) described in Manufacturing Example 39-2-1 and N-methylpyrrolidinone (1.5 mL) were added tri o-tolylphosphine (17 mg, 0.057 mmol) and palladium (II) 55 acetate (3.2 mg, 0.014 mmol) at room temperature, which was stirred for 5 hours at 120°C. The reaction mixture was cooled to room temperature, and filtered after addition of an aqueous potassium fluoride solution and ethyl acetate. The organic layer was separated, washed with Saturated aqueous sodium 60 To a mixture of 2,2-dimethyl-N-pyridin-2-yl-propiona chloride, and the solvent was evaporated under a reduced mide (3.0 g, 17 mmol) described in Manufacturing Example pressure. The residue was purified by reverse-phase high 39-1-1, N.N.N',N'-tetramethylethylenediamine (6.3 mL, 42 performance liquid chromatography (using an acetonitrile mmol) and tetrahydrofuran (60 mL) was added dropwise water mobile phase containing 0.1% trifluoroacetic acid) to n-butyl lithium (1.6 Mn-hexane solution, 30 mL, 47 mmol) at obtain the title compound (2.6 mg, 4%) as a trifluoroacetic 65 -78°C., which was stirred overnight at 0°C. Iodine (6.8 g. 27 acid salt. mmol) was added to the reaction mixture at -78°C., and MS m/e (ESI) 357.18 (MH) stirred for 1.5 hours at 0° C. Water and saturated aqueous US 8,158,657 B2 155 156 sodium thiosulfate solution were added to the reaction mix hydroxide solution (1 mL) was added. The resulting solids ture, which was then extracted with ethyl acetate. The organic were filtered to obtain the title compound (100 mg, 72%). layer was washed with Saturated aqueous Sodium chloride, 'H-NMR Spectrum (DMSO-d) & (ppm): 6.34 (2H, brs), and the solvent was evaporated under a reduced pressure. The 6.69 (1H, dd, J=4.8, 7.7 Hz), 7.62 (1H, dd, J=1.7, 7.7 Hz), residue was purified by silica gel column chromatography 5 7.90 (1H, s), 8.02 (1H, dd, J=1.7, 4.8 Hz), 8.45 (1H, s). (ethyl acetate:heptane=2:1) to obtain the title compound (2.9 The starting material, (4-benzyloxy-benzyl)-tributyl-stan g, 57%). nane, was synthesized as follows. 'H-NMR Spectrum (CDC1) & (ppm): 1.38 (9H, s), 6.85 Manufacturing Example 39-2-1 (1H, dd, J=4.8, 7.9 Hz), 7.94 (1H, brs), 8.11 (1H, dd, J=1.7, 10 7.9 Hz), 8.46 (1H, dd, J=1.7, 4.6 Hz). (4-Benzyloxy-benzyl)-tributyl-stannane Manufacturing Example 39-1-3 N-(3-(4-Bromo-pyrazol-1-yl)-pyridin-2-yl)-2,2-dim ethyl-propionamide 15

Br O e To a mixture of diisopropylamine (1.1 mL, 7.7 mmol) and N-NN ^ tetrahydrofuran (20 mL) was added dropwise n-butyl lithium 25 (1.6 M n-hexane solution, 4.5 mL, 7.1 mmol) at -78°C., 21 N. which was stirred for 30 minutes at that temperature. Tribu tyltin hydride (1.7 mL, 6.5 mmol) was added dropwise to the reaction mixture at the same temperature and then stirred for O 30 minutes at 0°C. The reaction mixture was cooled to -78° 30 C., and a mixture of 4-benzyloxybenzyl chloride (1.5 g. 6.5 mmol) and tetrahydrofuran (10 mL) was added dropwise at To a mixture of N-(3-iodo-pyridin-2-yl)-2,2-dimethyl-pro that temperature. The reaction mixture was gradually warmed pionamide (380 mg, 1.2 mmol) described in Manufacturing to room temperature. The reaction mixture was partitioned Example 39-1-2 and toluene (10 mL) were added 4-bromopy into water and n-heptane. The organic layer was washed with razole (160 mg, 1.1 mmol), copper (I) iodide (11 mg, 0.056 35 saturated aqueous Sodium chloride, and the Solvent was mmol), trans-1,2-cyclohexanediamine (26 mg, 0.22 mmol) evaporated under a reduced pressure. The residue was puri and potassium carbonate (340 mg, 2.5 mmol) at room tem fied by neutral silica gel column chromatography (ethyl perature, which was stirred overnight at 110°C. The reaction acetate:heptane=1:30) to obtain the title compound (2.6 g. mixture was concentrated under a reduced pressure. The resi 83%). due was purified by NH Silica gel column chromatography 40 H-NMR Spectrum (CDC1) & (ppm): 0.77-0.81 (6H, m), (ethyl acetate:heptane=2:1) to obtain the title compound (190 0.86 (9H, t, J–7.3 Hz), 1.21-1.30 (6H, m), 1.38-146 (6H, m), mg, 52%). 2.24 (2H, s), 5.01 (2H, s), 6.80-6.83 (2H, m), 6.88-6.91 (2H, H-NMR Spectrum (DMSO-d) & (ppm): 1.10 (9H, s), m), 7.29-7.44 (5H, m). 7.45 (1H, dd, J–4.8, 8.1 Hz), 7.84 (1H, s), 8.00 (1H, dd, J=1.7, Example 40 7.9 Hz), 8.23 (1H, s), 8.47 (1H, dd, J=1.7, 4.8 Hz), 9.83 (1H, 45 brs). 3-(3-(6-Phenoxy-pyridin-3-ylmethyl)-isoxazol-5-yl)- Manufacturing Example 39-1-4 pyridin-2,6-diamine

3-(4-Bromo-pyrazol-1-yl)-pyridin-2-ylamine 50

Br 55 e S-NN / 4. NH2 60

A mixture of N-(3-(4-bromo-pyrazol-1-yl)-pyridin-2-yl)- To a solution of (2-phenoxy-pyridin-5-yl)-acetohydroxi 2,2-dimethyl-propionamide (380 mg, 1.2 mmol) described in moyl chloride (59.1 mg, 225umol) described in Manufactur Manufacturing Example 39-1-3 and aqueous 2.5 N hydro 65 ing Example 40-1-4 and 3-ethynyl-pyridin-2,6-diamine (20.0 chloric acid solution (2 mL) was stirred overnight at 105°C. mg, 150 umol) described in Manufacturing Example 13-1-3 The reaction mixture was cooled to 0°C., and 5 N sodium in tetrahydrofuran (1.3 mL) was added triethylamine (41.8 US 8,158,657 B2 157 158 uL, 300 umol) at room temperature, which was stirred for 65 added to this reaction solution at -78°C., which was stirred minutes at 50° C. The reaction solution was allowed to room for further 10 minutes at room temperature. After allowing to temperature and partitioned into water and ethyl acetate. The room temperature, the reaction solution was partitioned into organic layer was separated, washed with Saturated aqueous water and ethyl acetate. The organic layer was separated, Sodium chloride, dried over anhydrous magnesium Sulfate washed with Saturated aqueous sodium chloride, dried over and filtered. The filtrate was concentrated under a reduced anhydrous magnesium sulfate and filtered. The filtrate was pressure. The residue was purified by NH silica gel column concentrated under a reduced pressure. The residue was puri chromatography (ethyl acetate:methanol=10:1) to obtain the fied by NH silica gel column chromatography (heptane:ethyl title compound (52 mg, 97%). acetate=5:1) to obtain the title compound (1.12g, 37%). H-NMR Spectrum (DMSO-d) & (ppm): 3.93 (2H, s), 10 H-NMR Spectrum (CDC1) & (ppm): 7.04 (1H, d. J=8.6 5.79 (2H, s), 5.81 (1H, d, J=8.4 Hz), 6.10 (2H, s), 6.40 (1H, s), Hz), 7.17 (2H, d, J=7.5 Hz), 7.26-7.31 (1H, m), 7.44-7.48 6.97 (1H, d, J=8.4 Hz), 7.08-7.10 (2H, m), 7.16-7.20(1H, m), (2H, m), 8.19 (1H, dd, J=2.2, 8.6 Hz), 8.63 (1H, d, J=2.2 Hz), 737-7.41 (2H, m), 7.50 (1H, d, J=8.4 Hz), 7.76 (1H, dd, 9.99 (1H, s). J=2.2, 8.4 Hz), 8.11 (1H, d. J=2.4 Hz). The starting material. (2-phenoxy-pyridin-5-yl)-acetohy 15 Manufacturing Example 40-1-3 droximoyl chloride, was synthesized as follows. 5-(2-Nitro-ethyl)-2-phenoxy-pyridine Manufacturing Example 40-1-1 5-Bromo-2-phenoxy-pyridine

N 25 2 N O 2 CuON O To a solution of 6-phenoxy-pyridine-3-carbaldehyde (1.12 To a solution of phenol (1.97 g. 20.9 mmol) in N,N-dim 30 g, 5.62 mmol) described in Manufacturing Example 40-1-2 in ethylformamide (100 mL) was added sodium hydride (1.00 g, acetic acid (10 mL) were added nitromethane (1.52 mL, 28.1 20.9 mmol) at 0°C., which was stirred for 5 minutes at 0°C. mmol) and ammonium acetate (86.6 mg, 11.2 mmol) under 2,5-Dibromopyridine (4.50 g. 19.0 mmol) was then added to nitrogen atmosphere, which was stirred for 3 hours at 100° C. this reaction solution at 0°C., and stirred for 40 minutes at After being cooled to room temperature, the reaction Solution room temperature. The reaction solution was then stirred for 35 was partitioned into water and ethylacetate. The organic layer further 3 hours at 120°C. After allowing to room temperature, was separated, washed with Saturated aqueous Sodium chlo the reaction Solution was partitioned into water and ethyl ride, dried over anhydrous magnesium sulfate and filtered. acetate. The organic layer was separated, washed with water The filtrate was concentrated under a reduced pressure. The and Saturated aqueous Sodium chloride, dried over anhydrous resulting residue was dissolved in dimethylsulfoxide (17 mL) magnesium sulfate and filtered. The filtrate was concentrated 40 and acetic acid (3 mL). Sodium borohydride (336 mg, 8.43 under a reduced pressure. The residue was purified by silica mmol) was added to this solution at room temperature while gel column chromatography (heptane:ethyl acetate 6: 1) to cooling appropriately, and stirred for 30 minutes at room obtain the title compound (3.85 g, 81%). temperature. The reaction solution was partitioned by addi H-NMR Spectrum (DMSO-d) & (ppm): 7.02 (1H, dd, tion of Sodium hydrogencarbonate, water and ethyl acetate. J=0.55, 8.8 Hz), 7.11-7.14 (2H, m), 7.19-7.23 (1H, m), 7.38 45 The organic layer was separated, washed with water and 7.43 (2H, m), 8.04 (1H, dd, J=2.6, 8.8 Hz), 8.25 (1H, dd, saturated aqueous sodium chloride, dried over anhydrous J=0.55, 2.6 Hz). magnesium sulfate and filtered. The filtrate was concentrated under a reduced pressure. The residue was purified by NH Manufacturing Example 40-1-2 silica gel column chromatography (heptane:ethyl acetate-3: 50 1) to obtain the title compound (753 mg, 55%). 6-Phenoxy-pyridine-3-carbaldehyde H-NMR Spectrum (CDC1) & (ppm): 3.28 (2H, t, J=7.1 Hz), 4.60 (2H, t, J=7.1 Hz), 6.88 (1H, d, J=8.8 Hz), 7.11-7.14 (2H, m), 7.20-7.24 (1H, m), 7.39-7.43 (2H, m), 7.55 (1H, ddd, J=0.37, 2.6, 8.4 Hz), 8.07 (1H, d, J=2.4 Hz). 55 Manufacturing Example 40-1-4 (2-Phenoxy-pyridin-5-yl)-acetohydroximoyl chloride 2 N O 60 To a solution of 5-bromo-2-phenoxy-pyridine (3.85g. 15.4 mmol) described in Manufacturing Example 40-1-1 in tet HO1 N rahydrofuran (60 mL) was added n-butyl lithium (10.6 mL, C 1.60 M hexane solution, 16.9 mmol) under nitrogen atmo 65 sphere at -78°C., which was stirred for 35 minutes at -78°C. N,N-Dimethylformamide (1.55 mL, 20.0 mmol) was then US 8,158,657 B2 159 160 To a solution of 5-(2-nitro-ethyl)-2-phenoxy-pyridine (753 7.86 (1H, dd, J=2.0, 7.6 Hz), 8.08 (1H, dd, J=2.0, 4.8 Hz), mg, 3.08 mmol) described in Manufacturing Example 40-1-3 8.57 (1H, d, J=3.2 Hz). in methanol (10 mL) was added lithium methoxide (234 mg. 2-Chloromethyl-5-fluoro-pyridine was synthesized as fol 6.16 mmol), which was stirred for 90 minutes at room tem lows. perature. The reaction solution was concentrated under a 5 reduced pressure. The resulting residue was suspended in a Manufacturing Example 41-1-1 mixture solution of tetrahydrofuran (10 mL) and methylene chloride (10 mL). Titanium (IV) chloride (745 uL, 6.87 mmol) was added to the Suspension under nitrogen atmo (5-Fluoro-pyridin-2-yl)-methanol sphere at -78°C., and stirred for 140 minutes at 0°C. The 10 reaction Solution was partitioned into water and ethyl acetate at 0°C. The organic layer was separated, washed with satu N rated aqueous sodium chloride, dried over anhydrous magne HO n sium sulfate and filtered. The filtrate was concentrated under 15 a reduced pressure to obtain the title compound (785 mg. 21 F 97%) as a crude product. H-NMR Spectrum (DMSO-d) & (ppm): 3.81 (2H, s), 6.99 (1H, dd, J=0.73, 8.4 Hz), 7.09-7.12 (2H, m), 7.17-7.21 To a solution of 2-bromo-5-fluoropyridine (3.67 g. 20.8 (1H, m), 7.38-7.42 (2H, m), 7.72 (1H, dd, J=2.6, 8.4 Hz), 8.03 mmol) in toluene (100 mL) was added dropwise n-butyl lithium (15.6 mL, 1.6M hexane solution, 25.0 mmol) under (1H, dd, J=0.55, 2.6 Hz), 11.8 (1H, s). nitrogen atmosphere at -78° C., which was stirred for 30 minutes. N,N-Dimethylformamide (8.05 mL, 104.0 mmol) Example 41 was added dropwise to this solution at -78°C., and stirred for 25 20 minutes at 0°C. This reaction solution was vigorously stirred after addition of water and tetrahydrofuran. The 3-(3-(4-(5-Fluoro-pyridin-2-ylmethoxy)-benzyl)- organic layer was separated, washed with water and Saturated isoxazol-5-yl)-pyridin-2-ylamine aqueous sodium chloride, dried over anhydrous magnesium sulfate and filtered. Sodium borohydride (1.58g, 41.8 mmol) 30 was added to the filtrate at 0°C., and stirred for 1 hour at room temperature. This reaction Solution was partitioned by addi tion of water and tetrahydrofuran. The organic layer was

separated, washed with Saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate and filtered. The 35 filtrate was concentrated under a reduced pressure. The resulting residue was purified by NH silica gel column chro matography (hexane:diethyl ether 1:2) to obtain the title compound (945mg, 36%). H-NMR Spectrum (CDC1) & (ppm): 4.75 (2H, s), 7.29 40 (1H, dd, J=4.4, 8.8 Hz), 7.43 (1H, ddd, J=2.8, 8.4, 8.4 Hz), 8.42 (1H, d. J=2.8 Hz). Manufacturing Example 41-1-2 45 2-Chloromethyl-5-fluoro-pyridine Tetrahydrofuran (10 mL) and 5 Naqueous sodium hydrox ide solution (448 uL., 2.24 mmol) were added to 4-(5-(2- amino-pyridin-3-yl)-isoxazol-3-ylmethyl)-phenol (600 mg. 2.24 mmol) described in Manufacturing Example 5-1-1, 50 N which was irradiated by ultrasonic wave for 1 minute. The C n reaction Solution was then concentrated under a reduced pres sure to obtain a white solid. 2-Chloromethyl-5-fluoro-pyri 2 F dine (359 mg, 2.46 mol) described in Manufacturing Example 41-2 and N,N-dimethylformamide (10 mL) were 55 added to the resulting white solid and stirred for 1 hour at 60° To a solution of (5-fluoro-pyridin-2-yl)-methanol (945mg, C. After being cooled to room temperature, the reaction solu 7.43 mmol) described in Manufacturing Example 41-1-1 in tion was partitioned into water and ethyl acetate. The organic methylene chloride (70 mL) was added dropwise thionyl layer was separated and concentrated under a reduced pres chloride (813 uL, 11.1 mmol) at room temperature, which 60 was stirred for 30 minutes. This reaction solution was parti sure. The residue was purified by NH silica gel column chro tioned by addition of water, Sodium hydrogencarbonate and matography (heptane:ethyl acetate=1:1) to obtain the title methylene chloride. The organic layer was separated, washed compound (650 mg, 77%). with Saturated aqueous sodium chloride, dried over anhy H-NMR Spectrum (DMSO-d) & (ppm): 3.96 (2H, s), drous magnesium sulfate and filtered. The filtrate was con 5.15 (2H, s), 6.25 (2H, brs), 6.69 (1H, dd, J–4.8, 8.0 Hz), 6.79 65 centrated under a reduced pressure. The residue was purified (1H, s), 6.99 (2H, d, J=8.4 Hz), 7.25 (2H, d, J=8.8 Hz), 7.59 by NH Silica gel column chromatography (hexane:diethyl (1H, dd, J=4.8, 8.8 Hz), 7.76 (1H, ddd, J=2.8, 8.8, 8.8 Hz), ether=1:1) to obtain the title compound (761.1 mg, 70%). US 8,158,657 B2 161 162 H-NMR Spectrum (CDC1) & (ppm): 4.67 (2H, s), 7.26 A mixed solution of (5-methyl-pyridine-2-yl)-methanol 7.51 (2H, m), 8.43 (1H, d, J=2.8 Hz). (500 mg. 4.1 mmol) described in Manufacturing Example 11-1-1, thionyl chloride (0.59 mL, 8.1 mmol) and methylene Example 42 chloride (10 mL) was stirred for 5 minutes under reflux. The reaction solution was cooled to room temperature and con 3-(3-(4-(5-Methyl-pyridin-2-ylmethoxy)-benzyl)- centrated under a reduced pressure. The resulting residue was partitioned into diethyl ether and saturated sodium bicarbon isoxazol-5-yl)-pyridin-2-ylamine ate Solution. The organic layer was separated and passed through a glass filter lined with silica gel (eluted with ethyl acetate). The eluate was concentrated to obtain the title com 10 pound (440 mg, 76%) as a crude product. The resulting com pound was used in the following reaction without further purification. Example 43 15 3-(3-(4-(4-Methyl-pyridin-2-yloxymethyl)-benzyl)- isoxazol-5-yl)-pyridin-2-ylamine

4-(5-(2-Amino-pyridin-3-yl)isoxazol-3-ylmethyl)-phenol (50 mg, 0.19 mmol) described in Manufacturing Example 5-1-1 and the 2-chloromethyl-5-methyl-pyridine (32 mg, 0.23 mmol) described in Manufacturing Example 42-1-2 25 were used to obtain the title compound (23 mg, 33%) accord ing to the methods similar to those of Example 10. 'H-NMR Spectrum (DMSO-d) & (ppm): 2.29 (3H, s), 3.95 (2H, s), 5.11 (2H, s), 6.25 (2H, brs), 6.69 (1H, dd, J=4.8, 30 8.0 Hz), 6.79 (1H, s), 6.97 (2H, d. J=8.4 Hz), 7.24 (2H, d, To a tetrahydrofuran (7.00 mL) solution of (4-(4-methyl J=8.4 Hz), 7.38 (1H, d, J=8.0 Hz), 7.62 (1H, d, J=8.0 Hz), pyridin-2-yloxymethyl)-phenyl)-acetohydroximoyl chloride 7.86 (1H, dd, J=1.6, 8.0 Hz), 8.08 (1H, dd, J=1.6, 4.8 Hz), (270 mg. 0.930 mmol) described in Manufacturing Example 8.40 (1H, s). 43-1-5 and 3-ethynyl-pyridin-2-ylamine (40.0 mg, 0.339 The starting material, 2-chloromethyl-5-methyl-pyridine mmol) described in Manufacturing Example 1-2-3 was added was synthesized as follows. 35 triethylamine (189 uL. 1.36 mmol) at room temperature, which was stirred at room temperature for 4 hours. Water was Manufacturing Example 42-1-1 added to the reaction solution at room temperature, which was then extracted with ethyl acetate. The organic layer was (5-Methyl-pyridin-2-yl)-methanol separated, washed with Saturated aqueous sodium chloride, 40 dried over anhydrous magnesium sulfate, and filtered. The filtrate was evaporated under a reduced pressure, and the residue was purified by NH Silica gel column chromatogra phy (ethyl acetate:heptane=1:3-> 1:2) to obtain the title com e pound (28.9 mg, 20.6%). HO N h 45 'H-NMR Spectrum (DMSO-d) & (ppm): 2.26 (3H, s), N 4.03 (2H, s), 5.30 (2H, s), 6.25 (2H, brs), 6.68-6.70 (2H, m), 6.80 (1H, s), 6.81-6.82 (1H, m), 7.32 (2H, d, J=8.0 Hz), 7.39 (2H, d, J=8.0 Hz), 7.86-7.88 (1H, m), 8.00-8.02 (1H, m), The title compound (1.1 g) was obtained according to the 8.08-8.09 (1H, m). method described in Manufacturing Example 11-1-1 through The starting material, (4-(4-methyl-pyridin-2-yloxym Manufacturing Example 11-1-3. 50 ethyl)-phenyl)-acetohydroximoyl chloride, was synthesized 'H-NMR Spectrum (DMSO-d) & (ppm): 2.27 (3H, s), as follows. 4.45 (2H, d, J=5.6 Hz), 5.31 (1H, t, J=5.6 Hz), 7.34 (1H, d, Manufacturing Example 43-1-1 J=8.0 Hz), 7.59 (1H, dd, J=1.6, 8.0 Hz), 8.31 (1H, d, J=1.6 Hz). 55 2-(4-Bromo-benzyloxy)-4-methyl-pyridine Manufacturing Example 42-1-2

2-Chloromethyl-5-methyl-pyridine Br 60

C N h 65 US 8,158,657 B2 163 164 To a mixture of 4-bromobenzyl alcohol (4.54 g. 24.3 To an acetic acid (20.0 mL) solution of 4-(4-methyl-pyri mmol) and N,N-dimethylformamide (50.0 mL) were added din-2-yloxymethyl)-benzaldehyde (2.60 g, 11.5 mmol) sodium hydride (1.00g, 25.0 mmol. 60% in oil) was added at described in Manufacturing Example 43-1-2 were added 0° C. under nitrogen atmosphere, which was stirred for 50 nitromethane (3.50 g, 57.3 mmol) and ammonium acetate minutes at room temperature. 2-Fluoro-4-methylpyridine (1.76 g. 22.9 mmol) at room temperature under nitrogen (1.80 g, 16.2 mmol) was then added thereto at 0°C., and atmosphere, which was stirred for 4 hours at 100° C. Water stirred for 2 hours and 30 minutes at room temperature. Water and ethyl acetate were added to the reaction mixture, and the was added to the reaction Solution at room temperature, organic layer was extracted with ethyl acetate. The organic which was then extracted with ethyl acetate. The organic layer was separated, washed with water and Saturated aque layer was separated, washed with Saturated aqueous sodium ous sodium chloride, dried over anhydrous magnesium Sul chloride, and filtered. The filtrate was evaporated under a 10 fate, and filtered. The filtrate was concentrated under a reduced pressure, and the residue was purified by Silica gel reduced pressure to obtain the title compound (3.40 g) as a column chromatography (ethyl acetate:heptane-1:15) to crude product. obtain the title compound (2.65 g, 58.8%). 'H-NMR Spectrum (DMSO-d) & (ppm): 2.28 (3H, s), H-NMR Spectrum (CDC1) & (ppm): 2.28 (3H, s), 5.31 5.39 (2H, s), 6.75 (1H, m), 6.84-6.85 (1H, m), 7.50-7.53 (2H, (2H, s), 6.60-6.61 (1H, m), 6.69-6.71 (1H, m), 7.29-7.32 (2H, 15 m), 7.85-7.87 (2H, m), 8.00-8.02(1H, m), 8.13 (1H, d, J–13.6 m), 7.46-7.48 (2H, m), 8.00-8.01 (1H, m). Hz), 8.23 (1H, d, J–13.6 Hz). Manufacturing Example 43-1-2 Manufacturing Example 43-1-4 4-(4-Methyl-pyridin-2-yloxymethyl)-benzaldehyde 4-Methyl-2-(4-(2-nitro-ethyl)-benzyloxy)pyridine O f 2 N 25 O N n 2

30

To a tetrahydrofuran (150 mL) solution of 2-(4-bromo To a dimethyl sulfoxide (50 mL) solution of 4-methyl-2- benzyloxy)-4-methyl-pyridine (5.70g, 20.5 mmol) described (4-((E)-2-nitro-vinyl)-benzyloxy)-pyridine (3.10 g, 11.5 in Manufacturing Example 43-1-1 was added dropwise n-bu mmol) described in Manufacturing Example 43-1-3 and ace tyl lithium (2.67 M n-hexane solution, 9.21 mL. 24.6 mmol) 35 tic acid (3.10 mL) was added sodium borohydride (73.3 mg, on a dry ice-ethanol bath (-78° C.) under nitrogen atmo 18.4 mmol) at room temperature while cooling appropriately sphere, which was stirred for 20 minutes at -78° C. N.N- under nitrogen atmosphere, which was stirred for 10 minutes. dimethylformamide (3.16 mL, 41.0 mmol) was then added Water was then added dropwise into the reaction solution at dropwise thereto and stirred for 10 minutes at -78°C. The room temperature while cooling appropriately, and the reac reaction solution was allowed to room temperature, water was 40 tion mixture was extracted with ethyl acetate. The organic added, and the solution was extracted with ethyl acetate. The layer was separated, washed with water and Saturated aque organic layer was separated and washed with Saturated aque ous sodium chloride, dried over anhydrous magnesium Sul ous sodium chloride, and the solvent was evaporated under a fate, and filtered. The filtrate was concentrated under a reduced pressure. The residue was purified by silica gel col reduced pressure, and the residue was purified by NH silica umn chromatography (ethyl acetate:heptane-1:3) to obtain 45 gel column chromatography (ethyl acetate: heptane-1:5-> 1: the title compound (2.58 g, 55.4%). 2) to obtain the title compound (1.10g, 35.1%). H-NMR Spectrum (CDC1) & (ppm): 2.31 (3H, s), 5.45 'H-NMR Spectrum (DMSO-d) & (ppm): 2.27 (3H, s), (2H, s), 6.66-6.67 (1H, m), 6.72-6.74 (1H, m), 7.58-7.60 (2H, 3.22 (2H, t, J=6.8 Hz), 4.84 (2H, t, J=6.8 Hz), 5.29 (2H, s), m), 7.85-7.88 (2H, m), 8.00-8.01 (1H, m), 10.0 (1H, s). 6.69 (1H, s), 6.82 (1H, d, J=5.2 Hz), 7.27 (2H, d. J=8.0 Hz), Manufacturing Example 43-1-3 50 7.37 (2H, d, J=8.0 Hz), 8.02 (1H, d, J=5.2 Hz). 4-Methyl-2-(4-((E)-2-nitro-vinyl)-benzyloxy)-pyri Manufacturing Example 43-1-5 dine (4-(4-Methyl-pyridin-2-yloxymethyl)-phenyl)-aceto hydroximoyl chloride 55 f 2n-1N C 60 O N N O N n HO1 n 21 2 65 US 8,158,657 B2 165 166 To a methanol (10.0 mL) solution of 4-methyl-2-(4-(2- The starting material, (4-(5-methyl-pyridin-2-yloxym nitro-ethyl)benzyloxy)pyridine (500 mg, 1.84 mmol) ethyl)-phenyl)-acetohydroximoyl chloride, was synthesized described in Manufacturing Example 43-1-4 was added as follows. lithium methoxide (140 mg, 3.68 mmol) under nitrogen atmosphere at room temperature, which was stirred for 30 5 Manufacturing Example 44-1-1 minutes at room temperature. The solvent was evaporated from the reaction mixture under a reduced pressure, and 2-(4-Bromo-benzyloxy)-5-methyl-pyridine anhydrous dichloromethane (10.0 mL) and anhydrous tet rahydrofuran (5.00 mL) were added to the residue. Titanium (IV) chloride (667 u , 6.07 mmol) was added dropwise into 10 the reaction mixture on a dry ice-ethanol bath (-78°C.), and Br stirred for 45 minutes at 0°C. and then for 60 minutes at room temperature. Water, ethyl acetate and tetrahydrofuran were added to the reaction mixture on an ice bath (0°C.), and the organic layer was separated. The organic layer was washed 15 with water and Saturated aqueous sodium chloride, dried over anhydrous magnesium Sulfate, and filtered. The filtrate was concentrated under a reduced pressure to obtain the title com pound (409 mg, 76.5%) as a crude product. To an N,N-dimethylformamide (50.0 mL) solution of 4-bromobenzyl alcohol (4.54 g. 24.3 mmol) was added H-NMR Spectrum (DMSO-d) & (ppm): 2.27 (3H, s), sodium hydride (1.00g, 25.0 mmol. 60% in oil) under nitro 3.82 (2H, s), 5.31 (2H, s), 6.70 (1H, s), 6.82-6.84 (1H, m), gen atmosphere at 0°C., which was stirred for 30 minutes at 7.24-728 (2H, m), 7.39-7.41 (2H, m), 8.01-8.03 (1H, m), room temperature. 2-Fluoro-5-methylpyridine (1.80 g, 16.2 11.73 (1H, s). mmol) was then added at 0°C., and stirred for 5 hours at room 25 temperature. Water was added to the reaction solution at room temperature, which was then extracted with ethylacetate. The Example 44 organic layer was separated and washed with Saturated aque ous sodium chloride, and the solvent was evaporated under a 3-(3-(4-(5-Methyl-pyridin-2-yloxymethyl)-benzyl)- reduced pressure. The residue was purified by silica gel col isoxazol-5-yl)-pyridin-2-ylamine 30 umn chromatography (ethyl acetate: heptane=1:15) to obtain the title compound (2.67g, 59.3%). H-NMR Spectrum (CDC1) & (ppm): 2.24 (3H, s), 5.30

(2H, s), 6.70-6.72(1H, m), 7.31-7.33 (2H, m), 7.38-7.41 (1H, 35 m), 7.46-7.49 (2H, m), 7.95-7.96 (1H, m). Manufacturing Example 44-1-2

40 4-(5-Methyl-pyridin-2-yloxymethyl)-benzaldehyde

45 To a tetrahydrofuran (7.00 mL) solution of (4-(5-methyl pyridin-2-yloxymethyl)-phenyl)-acetohydroximoyl chloride (246 mg, 0.846 mmol) described in Manufacturing Example 44-1-5 and 3-ethynyl-pyridin-2-ylamine (40.0 mg, 0.339 50 mmol) described in Manufacturing Example 1-2-3 was added triethylamine (189 uL. 1.36 mmol) at room temperature, To a tetrahydrofuran (150 mL) solution of 2-(4-bromo which was stirred for 4 hours at room temperature. Water was benzyloxy)-5-methyl-pyridine (5.40g, 19.4 mmol) described added to the reaction Solution at room temperature, which 55 in Manufacturing Example 44-1-1 was added dropwise n-bu was then extracted with ethyl acetate. The organic layer was tyl lithium (2.67 M n-hexane solution, 8.73 mL, 23.3 mmol) separated, washed with Saturated aqueous Sodium chloride, on a dry ice-ethanol bath (-78° C.) under nitrogen atmo dried over anhydrous magnesium sulfate, and filtered. The sphere, which was stirred for 30 minutes at -78° C. N.N- filtrate was concentrated under a reduced pressure, and the dimethylformamide (2.99 mL, 38.8 mmol) was then added residue was purified by NH silica gel column chromatogra 60 dropwise thereto, which was stirred for 10 minutes at -78°C. phy (ethyl acetate:heptane=1:3-> 1:2) to obtain the title com Water was added to the reaction solution at room temperature, pound (21.3 mg, 16.9%). which was then extracted with ethyl acetate. The organic 'H-NMR Spectrum (DMSO-d) & (ppm): 2.20 (3H, s), layer was separated and washed with Saturated aqueous 4.03 (2H, s), 5.28 (2H, s), 6.25 (2H, brs), 6.68-6.71 (1H, m), Sodium chloride, and the solvent was evaporated under a 6.75-6.77 (1H, m), 6.81 (1H, s), 7.32 (2H, d, J=8.0 Hz), 7.39 65 reduced pressure. The residue was purified by silica gel col (2H, d, J=8.0 Hz), 7.52-7.55 (1H, m), 7.85-7.88 (1H, m), umn chromatography (ethyl acetate:heptane=1:6-> 1:4) to 7.96-7.97 (1H, m), 8.08-8.09 (1H, m). obtain the title compound (2.93 g. 66.5%). US 8,158,657 B2 167 168 H-NMR Spectrum (CDC1) & (ppm): 2.21 (3H, s), 5.41 H-NMR Spectrum (DMSO-d) & (ppm): 2.20 (3H, s), (2H, s), 6.72-6.74 (1H, m), 7.38-7.41 (1H, m), 7.56-7.58 (2H, 3.22 (2H, t, J=6.8 Hz), 4.84 (2H, t, J=6.8 Hz), 5.27 (2H, s), m), 7.83-7.85 (2H, m), 7.92-7.93 (1H, m), 9.97 (1H, s). 6.76-6.78 (1H, m), 7.27 (2H, d, J=8.0 Hz), 7.36 (2H, d, J=8.0 Hz), 7.52-7.55 (1H, m), 7.97-7.98 (1H, m). Manufacturing Example 44-1-3 5 5-Methyl-2-(4-(E)-2-nitro-vinyl)-benzyloxy)-pyri Manufacturing Example 44-1-5 dine (4-(5-Methyl-pyridin-2-yloxymethyl)-phenyl)-aceto hydroximoyl chloride 10

C

15 N O N HO1 n 21

To an acetic acid (20.0 mL) solution of 4-(5-methyl-pyri To a methanol (30 mL) solution of 5-methyl-2-(4-(2-nitro din-2-yloxymethyl)-benzaldehyde (2.93 g, 12.9 mmol) ethyl)-benzyloxy)pyridine (700 mg, 2.57 mmol) described in described in Manufacturing Example 44-1-2 were added Manufacturing Example 44-1-4 was added lithium methox nitromethane (3.94 g. 64.5 mmol) and ammonium acetate ide (195 mg, 5.14 mmol) under nitrogen atmosphere at room (1.99 g, 25.8 mmol) under nitrogen atmosphere at room tem 25 temperature, which was stirred for 30 minutes at room tem perature, which was stirred for 2.5 hours at 100° C. Water and perature. The reaction mixture was concentrated under a ethyl acetate were added to the reaction mixture, and the reduced pressure, and anhydrous dichloromethane (15.0 mL) organic layer was extracted with ethyl acetate. This organic and anhydrous tetrahydrofuran (10.0 mL) were added to the layer was washed with water and Saturated aqueous sodium residue. Titanium (IV) chloride (904 uL, 8.22 mmol) was chloride, dried over anhydrous magnesium sulfate, and fil 30 added dropwise into the reaction mixture on a dry ice-ethanol tered. The filtrate was concentrated under a reduced pressure bath (-78°C.), and then stirred for 45 minutes at room tem to obtain the title compound (3.50 g) as a crude product. perature. Water, ethyl acetate and tetrahydrofuran were added H-NMR Spectrum (DMSO-d) & (ppm): 2.21 (3H, s), to the reaction mixture, and the organic layer was extracted 5.38 (2H, s), 6.82-6.84 (1H, m), 7.52 (2H, d, J=8.4 Hz), with ethyl acetate. This organic layer was washed with water 7.55-7.58 (1H, m), 7.85 (2H, d, J=8.4 Hz), 7.96-7.97 (1H, m), 35 and Saturated aqueous Sodium chloride, dried over anhydrous 8.12 (1H, d. J=13.6 Hz), 8.22 (1H, d, J=13.6 Hz). magnesium Sulfate, and filtered. The filtrate was concentrated under a reduced pressure to obtain the title compound (569 Manufacturing Example 44-1-4 mg, 76.1%) as a raw product. 'H-NMR Spectrum (DMSO-d) & (ppm): 2.02 (3H, s), 5-Methyl-2-(4-(2-nitro-ethyl)-benzyloxy)pyridine 40 3.81 (2H, s), 5.29 (2H, s), 6.77-6.79 (1H, m), 7.25 (2H, d, J=8.0 Hz), 7.40 (2H, d, J=8.0 Hz), 7.53-7.55 (1H, m), 7.97 7.98 (1H, m), 11.74 (1H, s). Example 45 45 3-(3-(4-(6-Fluoro-pyridin-2-ylmethoxy)-benzyl)- isoxazol-5-yl)-pyridin-2-ylamine

50

To a dimethylsulfoxide (40.0 mL) solution of 5-methyl-2- (4-((E)-2-nitro-vinyl)-benzyloxy)-pyridine (3.50 g, 12.9 mmol) described in Manufacturing Example 44-1-3 and ace 55 tic acid (3.50 mL) was added sodium borohydride (822 mg, 20.6 mmol) at room temperature while cooling appropriately under nitrogen atmosphere, which was stirred for 10 minutes. Water was then added dropwise at room temperature while cooling appropriately. The reaction mixture was extracted 60 with ethyl acetate, and the organic layer was washed with water and Saturated aqueous Sodium chloride, dried over Tetrahydrofuran (3 mL) and a 5 Naqueous sodium hydrox anhydrous magnesium Sulfate, and filtered. The filtrate was ide solution (36.0 uL, 0.18 mmol) were added to 4-(5-(2- concentrated under a reduced pressure, and the residue was amino-pyridin-3-yl)-isoxazol-3-ylmethyl)-phenol (48.2 mg, purified by NH silica gel column chromatography (ethyl 65 0.18 mmol) described in Manufacturing Example 5-1-1, acetate:heptane=1:4) to obtain the title compound (1.91 g, which was dissolved by irradiating ultrasonic wave for 1 54.3%). minute. The reaction mixture was then concentrated under a US 8,158,657 B2 169 170 reduced pressure to obtain a white solid. This solid and Manufacturing Example 46-1-6 and tetrahydrofuran (1 mL) 2-chloromethyl-6-fluoro-pyridine (63.2 mg, 0.43 mmol) were added 3-ethynyl-pyridin-2-ylamine (4.0 mg, 0.034 described in Manufacturing Example 45-1-1 were added to mmol) described in Manufacturing Example 1-2-3 and tri N,N-dimethylformamide (3 mL), which was stirred for 3 ethylamine (9.4 uL, 0.068 mmol) at room temperature, which hours at room temperature. This mixture was partitioned into was stirred for 3 hours at 45° C. The reaction mixture was water and ethyl acetate. This organic layer was separated, cooled to room temperature, water was added at the same washed with water and Saturated aqueous sodium chloride, temperature, and the mixture was extracted with ethylacetate. dried over anhydrous magnesium sulfate, and filtered. The The organic layer was washed with Saturated aqueous sodium filtrate was concentrated under a reduced pressure, and the chloride, and was concentrated under a reduced pressure. The 10 residue was purified by NH Silica gel column chromatogra residue was purified by NH silica gel column chromatogra phy (ethyl acetate:heptane=2:3) to obtain the title compound phy (heptane:ethyl acetate=1:1) to obtain the title compound (5.1 mg, 41%). (47.9 mg, 59%). H-NMR Spectrum (CDC1) & (ppm): 2.24 (3H, s), 3.90 H-NMR Spectrum (CDC1) & (ppm): 4.00 (2H, s), 5.12 (2H, s), 4.03 (2H, s), 5.53 (2H, brs), 5.85 (1H, d. J=2.9 Hz), (2H, s), 5.40 (2H, brs), 6.24 (1H, s), 6.71 (1H, dd, J–4.8, 7.6 15 5.87 (1H, d, J=2.9 Hz), 6.26 (1H, s), 6.72 (1H, dd, J=5.0, 7.6 Hz), 6.87 (1H, dd, J=2.8, 8.4 Hz), 6.94 (2H, d, J=8.8 Hz), 7.21 Hz), 7.21 (4H, s), 7.72 (1H, d. J–7.7 Hz), 8.12(1H, dd, J=1.8, (2H, d. J=8.8 Hz), 7.40-7.42(1H, m), 7.70 (1H, dd, J=1.6, 7.6 4.9 Hz). Hz), 7.81 (1H, q, J=8.0 Hz), 8.13 (1H, dd, J=1.6, 4.8 Hz) The starting material, (4-(5-methyl-furan-2-ylmethyl)- The starting material, 2-chloromethyl-6-fluoro-pyridine, phenyl)-acetohydroximoyl chloride, was synthesized as fol was synthesized as follows. lows. Manufacturing Example 45-1-1 Manufacturing Example 46-1-1 2-Chloromethyl-6-fluoro-pyridine 4-(Hydroxy-(5-methyl-furan-2-yl)-methyl)benzalde 25 hyde

C

30

A mixture of 2-fluoro-6-methylpyridine (420 mg, 3.78 mmol), N-chlorosuccimide (757 mg, 5.67 mmol), 75% ben Zoyl peroxide (24.4 mg., 0.08 mmol), acetic acid (13 uL, 0.23 35 mmol) and acetonitrile (7 mL) was stirred for 3 hours and 30 To a mixture of 4-bromobenzaldehyde dimethyl acetal (2.0 minutes at 85°C. The reaction mixture was cooled, water was mL, 12 mmol) and diethyl ether (30 mL) was added dropwise added, and the mixture was extracted with ethyl acetate. The n-butyl lithium (1.6 Mn-hexane solution, 9.0 mL, 14 mmol) organic layer was separated, washed with water and Saturated at -78° C., which was stirred for 20 minutes at the same aqueous sodium chloride, dried over anhydrous magnesium 40 temperature. 5-Methylfurfural (1.3 mL, 13 mmol) was added sulfate, and filtered. The filtrate was concentrated under a dropwise into the reaction mixture at that temperature, and reduced pressure, and the residue was purified by Silica gel stirred for 50 minutes at 0° C. Water and ethyl acetate were column chromatography (heptane:ethyl acetate-1: 1) to added to extract the reaction mixture. The organic layer was obtain the title compound (370.7 mg, 67%). washed with Saturated aqueous sodium chloride, and the Sol H-NMR Spectrum (DMSO-d) & (ppm): 4.75 (2H, s), 45 vent was evaporated under a reduced pressure. The residue 7.17-7.19 (1H, m), 7.50-7.52 (1H, m), 8.02-8.08 (1H, m). was purified by silica gel column chromatography (ethyl Example 46 acetate:heptane=1:2) to obtain the title compound (320 mg. 12%). 3-(3-(4-(5-Methyl-furan-2-ylmethyl)-benzyl)-isox H-NMR Spectrum (CDC1) & (ppm): 2.28 (3H, s), 5.86 azol-5-yl)-pyridin-2-ylamine 50 (1H, s), 5.90-5.91 (1H, m), 5.98 (1H, d, J=3.1 Hz), 7.63 (2H, d, J=8.4 Hz), 7.89 (2H, d, J=7.9 Hz), 10.03 (1H, s). Manufacturing Example 46-1-2

55 (4-(5-Methyl-furan-2-ylmethyl)-phenyl)-methanol

60 / O HO 21

65 To a mixture of lithium aluminumhydride (230 mg, 4.9 To a mixture of (4-(5-methyl-furan-2-ylmethyl)-phenyl)- mmol) and tetrahydrofuran (15 mL) was added aluminum acetohydroximoyl chloride (11 mg 0.043 mmol) described in chloride (830 mg, 6.2 mmol) at 0°C., which was stirred for 30 US 8,158,657 B2 171 172 minutes at room temperature. A mixture of 4-(hydroxy-(5- obtain the title compound (120 mg) as a crude product. This methyl-furan-2-yl)-methyl)-benzaldehyde (320 mg, 1.5 compound was used in the following reaction without further mmol) described in Manufacturing Example 46-1-1 and tet purification. rahydrofuran (5 mL) was added dropwise into the reaction mixture at 0°C., and stirred for 2 hours at that temperature. A Manufacturing Example 46-1-5 28% aqueous ammonia Solution was added dropwise into the reaction mixture at the same temperature to quench the excess 2-Methyl-5-(4-(2-nitro-ethyl)-benzyl)-furan reagent. The reaction mixture was cooled to room tempera ture, and filtered by being passed through a Celite bed. The filtrate was concentrated under a reduced pressure to obtain 10 the title compound (330 mg) as a crude product. This com pound was used in the following reaction without further Nt O purification. I 15 O - Manufacturing Example 46-1-3 To a mixture of 2-methyl-5-(4-((E)-2-nitro-vinyl)-benzyl)- 4-(5-Methyl-furan-2-ylmethyl)-benzaldehyde furan (120 mg) described in Manufacturing Example 46-1-4, acetic acid (0.2 mL) and dimethyl sulfoxide (3.4 mL) was added sodium borohydride (29 mg, 0.77 mmol) at room tem perature while cooling appropriately, which was stirred for 20 minutes at room temperature. Water was added to the reaction mixture, which was then extracted with ethyl acetate. The 25 organic layer was washed with Saturated aqueous sodium chloride, and was concentrated under a reduced pressure. The residue was purified by neutral silica gel column chromatog raphy (ethyl acetate:heptane=1:5) to obtain the title com pound (90 mg, 77%). To a mixture of (4-(5-methyl-furan-2-ylmethyl)-phenyl)- 30 methanol (350 mg, 1.7 mmol) obtained in Manufacturing H-NMR Spectrum (CDC1) & (ppm): 2.24 (3H, s), 3.30 Example 46-1-2 and dichloromethane (10 mL) was added (2H, t, J=7.4 Hz), 3.89 (2H, s), 4.59 (2H, t, J=7.4 Hz), 5.85 manganese dioxide (3.5 g, 4.7 mmol) at room temperature, 5.87 (2H, m), 7.14 (2H, d, J=8.2 Hz), 7.20 (2H, d, J=8.2 Hz). which was stirred overnight at room temperature. The reac tion mixture was filtered through a Celite pad, and the filtrate 35 Manufacturing Example 46-1-6 was concentrated under a reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate: (4-(5-Methyl-furan-2-ylmethyl)-phenyl)acetohy heptane=1:6) to obtain the title compound (100 mg, 29%). droximoyl chloride 'H-NMR Spectrum (CDC1) & (ppm): 2.25 (3H, s), 3.99 (2H, s), 5.876-5.883 (1H, m), 5.92 (1H, d, J=3.1 Hz), 7.39 40 7.41 (2H, m), 7.81-7.83 (2H, m), 9.99 (1H, s). C HO Manufacturing Example 46-1-4 O 45 W 21 2-Methyl-5-(4-((E)-2-nitro-vinyl)-benzyl)-furan To a mixture of 2-methyl-5-(4-(2-nitro-ethyl)-benzyl)-fu 50 ran (87 mg, 0.36 mmol) described in Manufacturing Example 46-1-5 and methanol (2 mL) was added lithium methoxide (27 mg, 0.71 mmol) at room temperature, which was stirred for 15 minutes at room temperature. The solvent was evapo rated from the reaction mixture under a reduced pressure. 55 Titanium(IV) chloride (86 uL, 0.78 mmol) was added at -78° C. to a mixture of the resulting residue, methylene chloride (2 mL) and tetrahydrofuran (1 mL), which was stirred for 1 hour To a mixture of 4-(5-methyl-furan-2-ylmethyl)-benzalde at 0°C. The reaction mixture was cooled to -78°C., water (5 hyde (96 mg, 0.48 mmol) described in Manufacturing mL) was added, and the temperature was gradually raised to Example 46-1-3 and acetic acid (1 mL) were added 60 room temperature. The reaction mixture was extracted by nitromethane (190 uL, 3.6 mmol) and ammonium acetate addition of ethyl acetate and water. The organic layer was (110 mg, 1.4 mmol) at room temperature, which was stirred washed with water until the pH became about 5. The organic for 3 hours at 100° C. The reaction mixture was cooled to layer was washed with Saturated aqueous Sodium chloride room temperature, and extracted by addition of water and and dried over anhydrous magnesium Sulfate. Thereafter, the ethyl acetate. The organic layer was washed with Saturated 65 organic layer was filtered and the filtrate was concentrated aqueous sodium chloride and dried over magnesium Sulfate. under a reduced pressure to obtain the title compound (79 mg, The filtrate was concentrated under a reduced pressure to 84%). US 8,158,657 B2 173 174 H-NMR Spectrum (CDC1) & (ppm): 2.24 (3H, s), 3.78 H-NMR Spectrum (DMSO-d) & (ppm): 2.45 (3H, s), (2H, s), 3.90 (2H, s), 5.85-5.87 (2H, m), 7.20 (4H, s). 4.50 (2H, d, J=5.2 Hz), 5.37 (1H, t, J=5.2 Hz), 7.11 (1H, d, J=5.2 Hz), 7.18 (1H, s), 8.36 (1H, d, J=5.2 Hz). Example 47 Example 48 3-(3-(4-(2-Methyl-pyridin-4-ylmethoxy)-benzyl)- isoxazol-5-yl)-pyridin-2-ylamine 3-(3-(5-p-Tolyloxy-thiophen-2-ylmethyl)-isoxazol-5- yl)-pyridin-2-ylamine

10

15

To a tetrahydrofuran (7.00 mL) solution of (5-p-tolyloxy (2-Methyl-pyridin-4-yl)-methanol (40 mg, 0.33 mmol) thiophen-2-yl)-acetohydroximoyl chloride (191 mg 0.678 described in Manufacturing Example 47-1-1, thionyl chlo 25 mmol) described in Manufacturing Example 48-1-5 and ride (0.047 ml, 0.65 mmol) and methylene chloride (4.0 ml) 3-ethynyl-pyridin-2-ylamine (40.0 mg, 0.339 mmol) were stirred for 5 minutes at 60° C. Sodium bicarbonate described in Manufacturing Example 1-2-3 was added tri Solution and ethyl acetate were added to separate the reaction ethylamine (189 uL. 1.36 mmol) at room temperature, which Solution, and the ethyl acetate layer was dried over sodium sulfate. The solvent was evaporated under a reduced pressure was stirred for 4 hours at room temperature. Water was added to obtain 4-chloromethyl-2-methyl-pyridine as a crude prod 30 to the reaction solution at room temperature, which was then uct. extracted with ethyl acetate. The organic layer was washed 2N Sodium hydroxide (0.16 ml, 0.32 mmol) and methanol with saturated aqueous sodium chloride and dried over anhy (1.0 ml) were added to dissolve 4-(5-(2-amino-pyridin-3-yl) drous magnesium sulfate, and the solvent was evaporated isoxazol-3-ylmethyl)-phenol (87 mg, 0.33 mmol) described under a reduced pressure. The residue was purified by NH in Manufacturing Example 5-1-1, and methanol was evapo 35 silica gel column chromatography (ethyl acetate: heptane=1: rated under a reduced pressure. A solution of the aforemen 3) to obtain the title compound (2.03 mg, 1.65%). tioned 4-chloromethyl-2-methyl-pyridine dissolved in dim 'H-NMR Spectrum (DMSO-d) & (ppm): 2.32 (3H, s), ethylformamide (1 ml) was added to the residue and stirred 4.14 (2H, s), 5.54 (2H, brs), 6.34-6.36 (1H, m), 6.40 (1H, s), for 10 minutes at 60° C. Water and ethyl acetate were added to 6.62-6.63 (1H, m), 6.73-6.77 (1H, m), 6.98-7.00 (2H, m), separate the reaction solution, the resulting ethyl acetate layer 40 7.11-7.13 (2H, m), 7.76-7.78 (1H, m), 8.14-8.15 (1H, m). was concentrated under a reduced pressure, and the residue The starting material. (5-p-tolyloxy-thiophen-2-yl)-aceto was purified by silica gel column chromatography (heptane: hydroximoyl chloride, was synthesized as follows. ethyl acetate=1:3) to obtain the title compound (47 mg,39%). 'H-NMR Spectrum (DMSO-d) & (ppm): 2.47 (3H, s), Manufacturing Example 48-1-1 3.96 (2H, s), 5.11 (2H, s), 6.25 (2H, brs), 6.68 (1H, dd, J=4.8, 45 8.0 Hz), 6.79 (1H, s), 6.97 (2H, d, J=8.8 Hz), 7.20 (1H, d, 5-p-Tolyloxy-thiophene-2-carbonitrile J=5.2 Hz), 7.25 (2H, d, J=8.8 Hz), 7.29 (1H, s), 7.86 (1H, dd, J=2.0, 8.0 Hz), 8.08 (1H, dd, J=2.0, 4.8 Hz), 8.42 (1H, d, J=5.2 Hz). The starting material. (2-methyl-pyridin-4-yl)-methanol, 50 was synthesized as follows. Manufacturing Example 47-1-1 (2-Methyl-pyridin-4-yl)-methanol 55 To a dimethyl sulfoxide (100 mL) solution of 5-nitro-2- thiophenecarbonitrile (6.30 g, 40.9 mmol) were added p-cresol (8.85g. 81.8 mmol) and potassium carbonate (11.3 e HO N 60 g, 81.8 mmol) under nitrogen atmosphere, which was stirred for 5 hours at 60°C. The reaction solution was cooled to room Null temperature, and extracted with ethyl acetate after addition of water. The organic layer was washed with Saturated aqueous Sodium chloride, and the solvent was evaporated under a The title compound (200 mg) was obtained according to 65 reduced pressure. The residue was purified by NH silica gel the methods similar to those of Manufacturing Example column chromatography (ethyl acetate: heptane=1:3->2:1) to 11-1-1 through Manufacturing Example 11-1-3. obtain the title compound (6.95 g, 78.9%). US 8,158,657 B2 175 176 H-NMR Spectrum (CDC1) & (ppm): 2.36 (3H, s), 6.38 H-NMR Spectrum (DMSO-d) & (ppm): 2.32 (3H, s), 6.39 (1H, m), 7.03-7.05 (2H, m), 7.18-7.20 (2H, m), 7.33 6.70 (1H, d, J=4.0 Hz), 7.18 (2H, d, J=8.0 Hz), 7.28 (2H, d, 7.35 (1H, m). J=8.0 Hz), 7.65 (1H, d, J=4.0 Hz), 7.78 (1H, d, J=12.8 Hz), 8.26 (1H, d. J=12.8 Hz). 5 Manufacturing Example 48-1-2 Manufacturing Example 48-1-4 5-p-Tolyloxy-thiophene-2-carbaldehyde 2-(2-Nitro-ethyl)-5-p-tolyloxy-thiophene

10

15

To a dimethyl sulfoxide (30.0 mL) solution of 2-((E)-2- To a tetrahydrofuran (70.0 mL) solution of 5-p-tolyloxy nitro-vinyl)-5-p-tolyloxy-thiophene (2.50 g, 9.57 mmol) thiophene-2-carbonitrile (2.00 g, 9.29. mmol) described in described in Manufacturing Example 48-1-3 and acetic acid (2.50 mL) was added sodium borohydride (610 mg, 20.6 Manufacturing Example 48-1-1 was added dropwise diisobu mmol) at room temperature while cooling appropriately tyl aluminum hydride (0.97 M n-hexane solution, 23.9 mL, 25 under nitrogen atmosphere, which was stirred for 30 minutes 23.2 mmol) on a dry ice-ethanol bath (-78°C.) under nitrogen at room temperature. Water was then added dropwise at room atmosphere, which was stirred for 3 hours at room tempera temperature while cooling appropriately. The reaction mix ture. The reaction mixture was cooled to room temperature ture was extracted with ethyl acetate, and the organic layer and extracted with ethyl acetate after addition of water. The was washed with water and Saturated aqueous sodium chlo organic layer was washed with Saturated aqueous sodium 30 ride, dried over anhydrous magnesium Sulfate, and filtered. chloride, and the solvent was evaporated under a reduced The filtrate was concentrated under a reduced pressure and pressure. The residue was purified by silica gel column chro the residue was purified by NH silica gel column chromatog matography (ethyl acetate:heptane=1:5) to obtain the title raphy (ethyl acetate:heptane=1:4) to obtain the title com compound (958 mg, 47.2%). pound (1.20 g, 47.6%). H-NMR Spectrum (CDC1) & (ppm): 2.36 (3H, s), 6.47 35 'H-NMR Spectrum (DMSO-d) & (ppm): 2.28 (3H, s), (1H, d, J=4.0 Hz), 7.08 (2H, d, J=8.0 Hz), 7.20 (2H, d, J=8.0 3.33 (2H, t, J=6.4 Hz), 4.81 (2H, t, J–6.4 Hz), 6.45-6.46 (1H, Hz), 7.51 (1H, d, J=4.0 Hz), 9.69 (1H, s). m), 6.67-6.69 (1H, m), 6.98-7.00 (2H, m), 7.17-7.20 (2H, m). Manufacturing Example 48-1-5 Manufacturing Example 48-1-3 40 (5-p-Tolyloxy-thiophen-2-yl)-acetohydroximoyl 2-(E)-2-Nitro-vinyl)-5-p-tolyloxy-thiophene chloride

45 C ry S 1 O HO1 N 50 To a methanol (10.0 mL) solution of 2-(2-nitro-ethyl)-5-p- tolyloxy-thiophene (500 mg, 1.90 mmol) described in Manu To an acetic acid (20.0 mL) solution of 5-p-tolyloxy facturing Example 48-1-4 was added lithium methoxide (144 55 mg, 3.80 mmol) under nitrogen atmosphere at room tempera thiophene-2-carbaldehyde (2.30 g, 10.5 mmol) described in ture, which was stirred for 30 minutes at room temperature. Manufacturing Example 48-1-2 were added nitromethane The solvent was evaporated from the reaction mixture under (3.20 g, 52.5 mmol) and ammonium acetate (1.62 g, 21.0 a reduced pressure, and anhydrous dichloromethane (15.0 mmol) under nitrogen atmosphere at room temperature, ml) and anhydrous tetrahydrofuran (10.0 ml) were added to which was stirred for 2.5 hours at 100° C. Water and ethyl 60 the residue. Titanium (IV) chloride (668 uL, 6.08 mmol) was acetate were added to the reaction mixture, and the organic added dropwise into the reaction mixture on a dry ice-ethanol layer was extracted with ethyl acetate. The organic layer was bath (-78° C.), which was stirred for 45 minutes at room washed with water and Saturated aqueous sodium chloride, temperature. Water, ethyl acetate and tetrahydrofuran were dried over anhydrous magnesium sulfate, and filtered. The added to the reaction mixture on an ice bath (0°C.), and the solvent was evaporated from the filtrate under a reduced 65 organic layer was extracted with ethyl acetate. This organic pressure to obtain the title compound (2.50 g) as a crude layer was washed with water and Saturated aqueous sodium product. chloride, dried over anhydrous magnesium sulfate, and fil US 8,158,657 B2 177 178 tered. The solvent was evaporated from the filtrate under a Example 50 reduced pressure to obtain the title compound (530 mg. 99.0%) as a crude product. 3-(3-(4-(Pyridin-3-ylmethoxy)-benzyl)-isoxazol-5- 'H-NMR Spectrum (DMSO-d) & (ppm): 2.28 (3H, s), yl)-pyridin-2-ylamine 3.94 (2H, s), 6.48 (1H, d. J=3.6 Hz), 6.74 (1H, d, J=3.6 Hz), 5 7.00-7.01 (2H, m), 7.18-7.20 (2H, m), 11.81 (1H, s).

Example 49 10 3-(3-(4-(Pyridin-4-ylmethoxy)-benzyl)-isoxazol-5- yl)-pyridin-2-ylamine

15

Tetrahydrofuran (3 mL) and a 5 Naqueous sodium hydrox ide solution (22.4 uL, 0.11 mmol) were added to 4-(5-(2- amino-pyridin-3-yl)-isoxazol-3-ylmethyl)-phenol (30 mg. 0.11 mmol) described in Manufacturing Example 5-1-1, which was dissolved by irradiating ultrasonic wave for 1 25 minute. Next, the reaction Solution was concentrated under a reduced pressure to obtain a white solid. This solid was sus pended in N,N-dimethylformamide (1 mL). Meanwhile, THF (390 ul) and a 1 Naqueous sodium hydroxide solution (390 mL, 0.39 mmol) were added to 3-(chloromethyl)pyridine 30 hydrochloride (50 mg, 0.39 mmol), and then the organic layer was separated to obtain a tetrahydrofuran solution of 3-(chlo romethyl)pyridine. A part of this tetrahydrofuran solution (224 uL) was added to the N,N-dimethylformamide suspen sion prepared above, and stirred for 45 minutes at 60°C. This 35 mixture was cooled to room temperature, and partitioned into water and ethyl acetate. The organic layer was separated, washed with water and Saturated aqueous sodium chloride, Tetrahydrofuran (3 mL) and a 5 Naqueous sodium hydrox dried over anhydrous magnesium sulfate, and filtered. The ide solution (22.4 uL, 0.11 mmol) were added to 4-(5-(2- filtrate was concentrated under a reduced pressure and the amino-pyridine-3-yl)-isoxazole-3-ylmethyl)-phenol (30 mg. 40 residue was purified by NH Silica gel column chromatogra 0.11 mmol) described in Manufacturing Example 5-1-1, phy (ethyl acetate) to obtain the title compound (40.0 mg. which was dissolved by irradiating ultrasonic wave for 1 100%). minute. Next, the reaction solution was concentrated under a 'H-NMR Spectrum (DMSO-d) & (ppm): 3.97 (2H, s), reduced pressure to obtain a white solid. The resulting solid 5.13 (2H, s), 6.25 (2H, brs), 6.67-6.74 (1H, m), 6.78 (1H, s), was suspended in N,N-dimethylformamide (1 mL). Mean 45 7.00 (2H, d, J=8.0Hz), 7.26 (2H, d, J=7.6Hz), 7.40-7.46 (1H, while, THF (390 LL) and 1 Naqueous sodium hydroxide m), 7.85-7.89 (2H, m), 8.09 (1H, d, J=4.8 Hz), 8.54 (1H, d, solution (390 uL, 0.39 mmol) were added to 4-(chloromethyl) J–4.8 Hz), 8.65-8.68 (1H, m). pyridine hydrochloride (50 mg, 0.39 mmol) and then the Example 51 organic layer was separated to obtain a tetrahydrofuran Solu 50 tion of 4-(chloromethyl)pyridine. A part of this tetrahydrofu 3-(3-(4-(4-Chloro-pyridin-2-ylmethoxy)-benzyl)- ran solution (224 pt) was added to the N,N-dimethylforma isoxazol-5-yl)-pyridin-2-ylamine mide Suspension prepared above, and stirred for 45 minutes at 60° C. This mixture was cooled to room temperature, and partitioned into water and ethyl acetate. The organic layer was 55 separated, washed with water and Saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate, and fil tered. The filtrate was concentrated under a reduced pressure and the residue was purified by NH silica gel column chro matography (ethyl acetate) to obtain the title compound (36 60 mg, 88%). 'H-NMR Spectrum (DMSO-d) & (ppm): 3.97 (2H, s), 5.17 (2H, s), 6.26 (2H, brs), 6.68-6.72 (1H, m), 6.79 (1H, s), 6.99 (2H, d, J=8.4 Hz), 7.26 (2H, d, J=8.8 Hz), 7.43 (2H, d, 65 J=6.0 Hz), 7.87 (1H, dd, J=2.0, 7.6 Hz), 8.09 (1H, dd, J=1.6, 4.8 Hz), 8.57 (2H, dd, J=1.6, 4.4 Hz). US 8,158,657 B2 179 180 Tetrahydrofuran (3 mL) and 5 Naqueous sodium hydrox H-NMR Spectrum (CDC1) & (ppm): 4.76 (2H, s), 7.23 ide solution (22.4 uL, 0.11 mmol) were added to 4-(5-(2- 7.25 (1H, m), 7.32-7.33 (1H, m), 8.46 (1H, d, J=5.6 Hz). amino-pyridine-3-yl)-isoxazole-3-ylmethyl)-phenol (30 mg. 0.11 mmol) described in Manufacturing Example 5-1-1, Manufacturing Example 51-1-2 which was dissolved by irradiating ultrasonic wave for 1 minute. Next, the reaction solution was concentrated under a 4-Chloro-2-chloromethyl-pyridine reduced pressure to obtain a white solid. An N,N-dimethyl formamide (1 mL) solution of 4-chloro-2-chloromethyl-py ridine (36.3 mg 0.22 mmol) described in Manufacturing 10 N Example 51-1-2 was added to a suspension of this solid and n C N,N-dimethylformamide (1 mL), which was stirred for 1 hour at 60°C. This mixture was cooled to room temperature, 2 and partitioned into water and ethyl acetate. The organic layer was separated, washed with water and Saturated aqueous 15 Sodium chloride, dried over anhydrous magnesium Sulfate, and filtered. The filtrate was concentrated under a reduced To a mixture of (4-chloro-pyridine-2-yl)-methanol (146.8 pressure and the residue was purified by NH silica gel column mg, 1.0 mmol) described in Manufacturing Example 51-1-1 chromatography (heptane:ethyl acetate 1:1) to obtain the and toluene (3 mL) was added thionyl chloride (112 u.L. 1.53 title compound (36.6 mg, 83%). mmol) on an ice bath, which was stirred for 1 hour 15 minutes H-NMR Spectrum (DMSO-d) & (ppm): 3.97 (2H, s), at room temperature. Saturated aqueous Sodium hydrogen 5.17 (2H, s), 6.26 (2H, brs), 6.69 (1H, dd, J–4.8, 8.0 Hz), 6.79 carbonate solution was added to the reaction mixture, which (1H, s), 7.01 (2H, d, J=8.4 Hz), 7.26 (2H, d, J=8.4 Hz), 7.51 was then extracted with ethyl acetate. The organic layer was (1H, dd, J=2.0, 5.2 Hz), 7.61 (1H, d, J=2.0 Hz), 7.87 (1H, dd, separated, washed with water and Saturated aqueous sodium J=2.0, 8.0 Hz), 8.08 (1H, dd, J=2.0, 4.8 Hz), 8.55 (1H, d, 25 chloride, dried over anhydrous magnesium sulfate, and fil J=5.2 Hz). tered. The filtrate was concentrated under a reduced pressure The starting material, 4-chloro-2-chloromethyl-pyridine, and the residue was purified by NH silica gel column chro was synthesized as follows. matography (heptane:ethyl acetate=4:1) to obtain the title 30 compound (97 mg, 59%). Manufacturing Example 51-1-1 H-NMR Spectrum (CDC1) & (ppm): 4.65 (2H, s), 7.26 7.28 (1H, m), 7.52-7.53 (1H, m), 8.48 (1H, d, J=5.6 Hz). (4-Chloro-pyridin-2-yl)-methanol Example 52 35 3-(3-(4-(6-Chloro-pyridin-2-ylmethoxy)-benzyl)- isoxazol-5-yl)-pyridin-2-ylamine

n OH

2 40

To a mixture of 4-chloro-2-picoline (1.0g, 7.84 mmol) and 45 dichloromethane (20 mL), was added m-chloroperbenzoic acid (3.5g, 13.2 mmol) on an ice bath, which was stirred for C 1.5 hours at room temperature. Water and Sodium hydrogen carbonate were added to the reaction, followed by extraction with dichloromethane. The organic layer was separated, 50 washed with water and Saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate, and filtered. Acetic Tetrahydrofuran (3 mL) and a 5 Naqueous sodium hydrox anhydride (20 mL) was added to the residue obtained by ide solution (22.4 uL, 0.11 mmol) were added to 4-(5-(2- concentrating the filtrate under a reduced pressure, and this amino-pyridin-3-yl)isoxazol-3-ylmethyl)-phenol (30 mg. was stirred for 1 hour at 100° C. The reaction mixture was 55 0.11 mmol) described in Manufacturing Example 5-1-1, cooled to room temperature and concentrated under a reduced which was dissolved by irradiating ultrasonic wave for 1 pressure. A 5 N aqueous sodium hydroxide solution (1.57 minute. The reaction Solution was then concentrated under a mL, 7.87 mmol) was added to a mixture of the resulting reduced pressure to obtain a white solid. An N,N-dimethyl residue and methanol (20 mL) on an ice bath, which was formamide (1 mL) solution of 2-chloro-6-chloromethyl-py stirred for 1.5 hours at room temperature. Water was added to 60 ridine (36.3 mg, 0.22 mmol) described in Manufacturing the mixture, which was then extracted with ethyl acetate. The Example 52-1-2 was added to a suspension of this solid and organic layer was separated, washed with water and Saturated N,N-dimethylformamide (1 mL), which was stirred for 1 aqueous sodium chloride, dried over anhydrous magnesium hour at 60°C. This mixture was cooled to room temperature sulfate, and filtered. The filtrate was concentrated under a and partitioned into water and ethyl acetate. The organic layer reduced pressure and the residue was purified by NH silica gel 65 was separated, washed with water and Saturated aqueous column chromatography (heptane:ethyl acetate-6:1) to Sodium chloride, dried over anhydrous magnesium sulfate, obtain the title compound (200 mg, 18%). and filtered. The filtrate was concentrated under a reduced US 8,158,657 B2 181 182 pressure, and the residue was purified by NH silica gel col washed with water and Saturated aqueous sodium chloride, umn chromatography (heptane:ethyl acetate-1: 1) to obtain dried over anhydrous magnesium sulfate, and filtered. The the title compound (39.5 mg, 90%). filtrate was concentrated under a reduced pressure, and the H-NMR Spectrum (DMSO-d) & (ppm): 3.97 (2H, s), residue was purified by NH Silica gel column chromatogra 5.15 (2H, s), 6.26 (2H, brs), 6.69 (1H, dd, J–4.8, 8.0 Hz), 6.79 phy (heptane:ethyl acetate-4:1) to obtain the title compound (1H, s), 6.99 (2H, d. J=8.4 Hz), 7.26 (2H, d, J=8.4 Hz), (163.2 mg, 73%). 7.46-7.52 (2H, m), 7.85-7.92 (2H, m), 8.08 (1H, dd, J=2.0, H-NMR Spectrum (CDC1) & (ppm): 4.64 (2H, s), 7.29 4.8 Hz). (1H, d, J=8.0Hz), 7.44 (1H, d, J=7.6Hz), 7.70 (1H, dd, J=7.6, The starting material, 2-chloro-6-chloromethyl-pyridine, 8.0 Hz). was synthesized as follows. 10 Example 53 Manufacturing Example 52-1-1 3-(3-(6-Phenoxy-pyridin-3-ylmethyl)-isoxazol-5-yl)- (6-Chloro-pyridin-2-yl)-methanol pyridin-2-ylamine 15

C N n OH 2

To a mixture of 2-chloro-6-methylpyridine (1.0 g, 7.84 mmol) and dichloromethane (20 mL) was added m-chlorop 25 erbenzoic acid (3.5g, 13.2 mmol) on an ice bath, which was stirred for 1.5 hours at 40° C. Water and sodium hydrogen carbonate were added to the reaction mixture, which was then extracted with dichloromethane. The organic layer was sepa To a tetrahydrofuran (2 mL) solution of (2-phenoxy-pyri rated, washed with water and Saturated aqueous sodium chlo 30 ride, dried over anhydrous magnesium Sulfate, and filtered. din-5-yl)-acetohydroximoyl chloride (100 mg 0.381 mmol) The filtrate was concentrated under a reduced pressure. Ace described in Manufacturing Example 40-1-4 and 3-ethynyl tic anhydride (20 mL) was added to the resulting residue and pyridin-2-ylamine (30 mg, 0.25 mmol) described in Manu stirred for 1 hour at 100° C. The reaction mixture was cooled facturing Example 1-2-3 was added triethylamine (714, 0.51 to room temperature and concentrated under a reduced pres 35 mmol) under nitrogen atmosphere, which was stirred for 3 sure. A 5 Naqueous sodium hydroxide solution (4 mL. 20.1 hours at 50° C. Water was added to the reaction mixture at mmol) was added to a mixture of the resulting residue and room temperature, which was then extracted with ethyl methanol (20 mL) on an ice bath, which was stirred for 30 acetate. The organic layer was separated, washed with water minutes. Water was added to this mixture, which was then and Saturated aqueous Sodium chloride, dried over anhydrous extracted with ethyl acetate. The organic layer was separated, 40 magnesium Sulfate, and filtered. The filtrate was concentrated washed with water and Saturated aqueous sodium chloride, under a reduced pressure, and the residue was purified by dried over anhydrous magnesium sulfate, and filtered. The silica gel column chromatography (ethyl acetate:metha filtrate was concentrated under a reduced pressure, and the nol=10:1) to obtain the title compound (27 mg, 31%). residue was purified by NH silica gel column chromatogra H-NMR Spectrum (DMSO-d) & (ppm): 4.02 (2H, s), phy (heptane:ethyl acetate=3:1) to obtain the title compound 45 6.26 (2H, s), 6.68 (1H, dd, J=4.8, 7.7 Hz), 6.83 (1H, s), 6.98 (200.0 mg, 18%). (1H, d, J=8.6 Hz), 7.09 (2H, d, J=7.5 Hz), 7.18 (1H, t, J=7.3 'H-NMR Spectrum (CDC1) & (ppm): 3.08 (1H, brs), 4.75 Hz), 7.39 (2H, t, J–7.5 Hz), 7.79 (1H, dd, J=2.4, 8.6 Hz), 7.85 (1H, dd, J=1.8, 7.7 Hz), 8.07 (1H, dd, J=1.8, 4.8 Hz), 8.13 (2H, d, J=5.2 Hz), 7.23-7.27 (2H, m), 7.64-7.69 (1H, m). (1H, d, J=2.2 Hz). Manufacturing Example 52-1-2 50 Example 54 2-Chloro-6-chloromethyl-pyridine 3-(3-(6-Phenoxymethyl-pyridin-3-ylmethyl)-isox azol-5-yl)-pyridin-2-ylamine

55 C N n C

2

60 To a mixture of (6-chloro-pyridine-2-yl)-methanol (200 mg, 1.39 mmol) described in Manufacturing Example 52-1-1 and toluene (3 mL) was added thionyl chloride (152 uL 2.09 mmol) on an ice bath, which was stirred for 2 hours at room temperature. Saturated aqueous sodium hydrogencarbonate 65 Solution was added to the reaction mixture, which was then extracted with ethyl acetate. The organic layer was separated, US 8,158,657 B2 183 184 To a tetrahydrofuran (3.00 mL) solution of (6-phenoxym Example 54-1-1 was added dropwise thionyl chloride (3.65 ethyl-pyridin-3-yl)acetohydroximoyl chloride (80.0 mg. mL, 50.1 mmol) on an ice bath (0°C.) under nitrogen atmo 0.289 mmol) described in Manufacturing Example 54-1-6 sphere, which was stirred for 5 minutes at room temperature. and 3-ethynyl-pyridin-2-ylamine (20.0 mg, 0.169 mmol) The solvent was evaporated under a reduced pressure to described in Manufacturing Example 1-2-3 was added tri obtain the title compound (4.2 g. 69.2%) as a hydrochloride. ethylamine (70.7 uL, 0.507 mmol) at room temperature, H-NMR Spectrum (DMSO-d) & (ppm): 4.78 (2H, s), which was stirred for 4.5 hours at 60° C. Water was added to 7.55-7.57 (1H, m), 8.11-8.14 (1H, m), 8.70-8.72 (1H, m). the reaction solution at room temperature, which was then extracted with ethyl acetate. The organic layer was washed Manufacturing Example 54-1-3 with Saturated aqueous Sodium chloride and dried over anhy 10 drous magnesium sulfate, and the solvent was evaporated 5-Bromo-2-phenoxymethyl-pyridine under a reduced pressure. The residue was purified by NH silica gel column chromatography (ethyl acetate: heptane 2: 1->3:1) to obtain the title compound (4.00 mg. 6.60%). H-NMR Spectrum (DMSO-d) & (ppm): 4.10 (2H, s), 15 Br 5.16 (2H, s), 6.27 (2H, brs), 6.69-6.72 (1H, m), 6.87 (1H, s), N 6.92-7.02 (3H, m), 7.27-7.31 (2H, m), 748-7.50 (1H, m), 7.78-7.79 (1H, m), 7.86-7.88 (1H, m), 8.09-8.10 (1H, m), 2 8.58-8.59 (1H, m). N The starting material, (6-phenoxymethyl-pyridin-3-yl)-ac etohydroximoyl chloride, was synthesized as follows. Manufacturing Example 54-1-1 To an N,N-dimethylformamide (40.0 mL) solution of phe nol (1.92g, 20.4 mmol) was added sodium hydride (815 mg. (5-Bromo-pyridin-2-yl)-methanol 25 20.4 mmol. 60% in oil) on an ice bath (0°C.) under nitrogen atmosphere, which was stirred for 20 minutes at room tem perature. To the reaction solution was then added a mixture of 5-bromo-2-chloromethyl-pyridine hydrochloride (4.2g, 20.4 Br N mmol) described in Manufacturing Example 54-1-2 and tri 30 ethylamine (28.0 mL. 20.4 mmol), which was stirred first for 2 OH 30 minutes at room temperature and then for 45 minutes at N 70° C. Water and ethyl acetate were added to the reaction mixture, and the organic layer was extracted with ethyl To a toluene (300 mL) solution of 2,5-dibromopyridine acetate. This organic layer was washed with water and Satu (10.0 g, 42.2 mmol) was added dropwise n-butyl lithium 35 rated aqueous sodium chloride, dried over anhydrous magne (2.55 M n-hexane solution, 18.2 mL, 46.4 mmol) on a dry sium sulfate, and filtered. The solvent was evaporated from ice-ethanol bath (-78°C.) under nitrogen atmosphere, which the filtrate under a reduced pressure, and the residue was was stirred for 2 hours at -78° C. N.N-dimethylformamide purified by silica gel column chromatography (ethyl acetate: (3.7 g, 50.6 mmol) was then added dropwise thereto and heptane=1:10) to obtain the title compound (4.40 g, 81.7%). stirred for 10 minutes at -78°C. Sodium borohydride (3.20g, 40 H-NMR Spectrum (CDC1) & (ppm): 5.15 (2H, s), 6.95 84.4 mmol) and methanol (20.0 mL) were then added and 6.99 (3H, m), 7.25-7.31 (2H, m), 7.42-7.45 (1H, m), 7.81 stirred for 30 minutes at room temperature. Water was added 7.83 (1H, m), 8.64-8.65 (1H, m). to the reaction solution, which was then extracted with ethyl Manufacturing Example 54-1-4 acetate. The organic layer was washed with Saturated aqueous Sodium chloride, and the solvent was evaporated under a 45 6-Phenoxymethyl-pyridin-3-carbaldehyde reduced pressure. The residue was purified by silica gel col umn chromatography (ethyl acetate:heptane=1:1->2:1) to obtain the title compound (4.70 g, 59.2%). H-NMR Spectrum (DMSO-d) & (ppm): 4.54 (2H, d, J=5.6 Hz), 5.28 (1H, t, J–5.6 Hz), 7.44-7.47 (1H, m), 8.03 50 8.05 (1H, m), 8.59-8.60 (1H, m).

Manufacturing Example 54-1-2 2 O 5-Bromo-2-chloromethyl-pyridine hydrochloride 55

Br To a diethyl ether (250 mL) solution of 5-bromo-2-phe N 60 noxymethyl-pyridine (4.40g, 16.6 mmol) described in Manu facturing Example 54-1-3 was added n-butyl lithium (2.55 M 2 C n-hexane solution, 8.46 mL. 21.6 mmol) on a dry ice-ethanol N bath (-78°C.) under nitrogen atmosphere, which was stirred H-Cl for 40 minutes at -78°C. N.N-dimethylformamide (1.93 mL, 65 25.0 mmol) was then added dropwise thereto and stirred for To a toluene (20.0 mL) solution of (5-bromo-pyridin-2-yl)- 20 minutes at -78°C. The reaction solution was allowed to methanol (4.70 g, 25.0 mmol) described in Manufacturing room temperature, water was added, and the Solution was US 8,158,657 B2 185 186 extracted with ethyl acetate. The organic layer was washed Manufacturing Example 54-1-6 with Saturated aqueous sodium chloride, and the solvent was evaporated under a reduced pressure. The residue was puri fied by silica gel column chromatography (ethyl acetate:hep (6-Phenoxymethyl-pyridin-3-yl)-acetohydroximoyl tane=1:3) to obtain the title compound (1.00g, 28.3%). chloride H-NMR Spectrum (CDC1) & (ppm): 5.29 (2H, s), 6.97 7.01 (3H, m), 7.29-7.33 (2H, m), 7.73-7.75 (1H, m), 8.19 8.21 (1H, m), 9.05-9.06 (1 m), 10.12 (1H, s). 10 Manufacturing Example 54-1-5

5-(2-Nitro-ethyl)-2-phenoxymethyl-pyridine 15 To a methanol (7.00 mL) solution of 5-(2-nitro-ethyl)-2- phenoxymethyl-pyridine (170 mg. 0.658 mmol) described in Manufacturing Example 54-1-5 was added lithium methox ide (50.0 mg, 1.32 mmol) under nitrogen atmosphere at room temperature, which was stirred for 30 minutes at room tem perature. The solvent was evaporated from the reaction mix ture under a reduced pressure, and anhydrous dichlo N romethane (10.0 ml) and anhydrous tetrahydrofuran (5.00 25 ml) were added to the residue. Titanium (IV) chloride (231 2 LL, 2.11 mmol) was added dropwise into the reaction mixture on a dry ice-ethanol bath (-78°C.), which was stirred for 30 minutes at room temperature. Water, ethyl acetate and tet rahydrofuran were added to the reaction mixture on an ice 30 bath (0°C.), and the organic layer was extracted with ethyl acetate. This organic layer was washed with water and Satu rated aqueous sodium chloride, dried over anhydrous magne sium sulfate, and filtered. The solvent was evaporated from the filtrate under a reduced pressure to obtain the title com To a methanol (20.0 mL) solution of 6-phenoxymethyl 35 pyridine-3-carbaldehyde (1.00 g, 4.69 mmol) described in pound (169 mg) as a crude product. Manufacturing Example 54-1-4 was added lithium methox H-NMR Spectrum (DMSO-d) & (ppm): 3.90 (2H, s), ide (21.4 mg., 0.56 mmol) under nitrogen atmosphere at room 5.17 (2H, s), 6.93-6.97 (1H, m), 701-703 (2H, m), 7.27-7.30 temperature. This was cooled to 0°C., and nitromethane (372 (2H, m), 7.49-7.51 (1H, m), 7.72-7.74 (1H, m), 8.49-8.50 mg, 6.10 mmol) and lithium methoxide (193 mg, 5.07 mmol) 40 (1H, m), 11.83 (1H, s). were added and stirred for 10 minutes at room temperature. The reaction solution was then concentrated under a reduced Example 55 pressure. Tetrahydrofuran (20.0 mL) was added to the resi due, and then acetic anhydride (6.24 g. 61.1 mmol) and tri 3-(3-(4-(6-Fluoro-pyridin-2-yloxymethyl)-benzyl)- ethylamine (1.42 mL, 10.2 mmol) were added and stirred for 45 isoxazol-5-yl)-pyridin-2-ylamine 1 hour at 70° C. Water and ethyl acetate were added to the reaction mixture, and the organic layer was extracted with ethyl acetate. This organic layer was washed with water and saturated aqueous sodium chloride, dried over anhydrous magnesium Sulfate, and filtered. The solvent was evaporated 50 from the filtrate under a reduced pressure. Methanol (20.0 mL) was added to the residue, and sodium borohydride (263 mg, 6.96 mmol) was then added on an ice bath (0° C.). Following 5 minutes of stirring at 0°C., water was added 55 dropwise at 0°C. The reaction mixture was extracted with ethyl acetate, and the organic layer was washed with water and Saturated aqueous Sodium chloride, dried over anhydrous magnesium Sulfate, and filtered. The solvent was evaporated from the filtrate under a reduced pressure, and the residue was purified by NH silica gel column chromatography (ethyl 60 acetate:heptane=1:1) to obtain the title compound (170 mg. To a tetrahydrofuran (3 mL) solution of (4-(6-fluoro-pyri 14.2%). din-2-yloxymethyl)-phenyl)acetohydroximoyl chloride (200 H-NMR Spectrum (DMSO-d) & (ppm): 3.25 (2H, t, mg, 0.679 mmol) described in Manufacturing Example J=6.8 Hz), 4.91 (2H, t, J=6.8 Hz), 5.14 (2H, s), 6.93-6.97 (1H, 65 55-1-5 and 3-ethynyl-pyridin-2-ylamine (50 mg, 0.423 m), 7.00-7.02 (2H, m), 7.27-7.31 (2H, m), 7.46-7.48 (1H, m), mmol) described in Manufacturing Example 1-2-3 was added 7.75-7.78 (1H, m), 8.49-8.50 (1H, m). triethylamine (237 uL. 1.7 mmol) at room temperature, which US 8,158,657 B2 187 188 was stirred for 2 hours at 50° C. Water was added to the chloride, dried over anhydrous magnesium sulfate, and fil reaction solution at room temperature, which was then tered. The filtrate was concentrated under a reduced pressure, extracted with ethyl acetate. The organic layer was washed and the residue was purified by silica gel column chromatog with Saturated aqueous Sodium chloride and dried over anhy raphy (heptane:ethyl acetate=10: 1-4:1) to obtain the title drous magnesium sulfate, and the solvent was evaporated compound (4.57 g, 75%). under a reduced pressure. The residue was purified by NH H-NMR Spectrum (CDC1) & (ppm): 5.43 (2H, s), 6.50 silica gel column chromatography (heptane:ethyl acetate-4: 6.53 (1H, m), 6.70-6.72 (1H, m), 7.60-7.62 (2H, m), 7.66 1->2:1) to obtain the title compound (59 mg, 23%). H-NMR Spectrum (CDC1) & (ppm): 4.07 (2H, s), 5.32 7.72 (1H, m), 7.88-7.91 (2H, m), 10.0 (1H, s). (2H, s), 5.64 (2H, brs), 6.27 (1H, s), 6.47-6.50 (1H, m), 10 6.64-6.67 (1H, m), 6.71-6.74 (1H, m), 7.30 (2H, d. J–8.4 Hz), Manufacturing Example 55-1-3 7.43 (2H, d, J=8.4 Hz), 7.63-7.69 (1H, m), 7.72-7.75 (1H, m), 8.11-8.12 (1H, m). 2-Fluoro-6-(4-(E)-2-nitro-vinyl)-benzyloxy)-pyri The starting material, (4-(6-fluoro-pyridin-2-yloxym dine ethyl)-phenyl)acetohydroximoyl chloride, was synthesized 15 as follows. Manufacturing Example 55-1-1 O e N f 4-(6-Fluoro-pyridin-2-yloxymethyl)-benzonitrile INt S. O N O F

25 A mixture of 4-(6-fluoro-pyridin-2-yloxymethyl)-benzal dehyde (4.57 g. 19.8 mmol) described in Manufacturing Example 55-1-2, nitromethane (2.13 mL, 39.6 mmol), ammonium acetate (2.29 g, 29.7 mmol) and acetic acid (45.7 mL) was stirred for 19 hours at 100° C. This mixture was To an N,N-dimethylformamide (50 mL) solution of 2.6- 30 cooled to room temperature and concentrated under a reduced difluoropyridine (5 g, 43.4 mmol) and 4-(hydroxymethyl) pressure. The residue was dissolved in ethyl acetate, washed benzonitrile (8.67 g. 65.1 mmol) was added sodium hydride with water and Saturated aqueous sodium chloride, dried over (2.56 g. 65.1 mmol. 60% in oil) at room temperature. This anhydrous magnesium Sulfate, and filtered. The filtrate was mixture was stirred for 4 hours at 70° C. This mixture was concentrated under a reduced pressure, and the residue was partitioned into ethylacetate and water. The organic layer was 35 purified by silica gel column chromatography (heptane:ethyl separated, washed with water, dried over anhydrous magne acetate-4:1) to obtain the title compound (3.44 g. 63%). sium sulfate, and filtered. The filtrate was concentrated under H-NMR Spectrum (CDC1) & (ppm): 5.39 (2H, s), 6.50 a reduced pressure, and the residue was purified by NH-silica 6.53 (1H, m), 6.68-6.71 (1H, m), 7.52-7.61 (5H, m), 7.66 gel column chromatography (heptane:ethyl acetate 10:1-4: 40 7.72 (1H, m), 8.03-8.99 (1H, m). 1) to obtain the title compound (5.99 g, 61%). H-NMR Spectrum (CDC1) & (ppm): 5.41 (2H, s), 6.74 6.77 (1H, m), 6.87-6.89 (1H, m), 7.63-7.66 (2H, m), 7.85 Manufacturing Example 55-1-4 7.88 (2H, m), 7.90-7.96 (1H, m). 2-Fluoro-6-(4-(2-nitro-ethyl)-benzyloxy)-pyridine Manufacturing Example 55-1-2 45 4-(6-Fluoro-pyridin-2-yloxymethyl)-benzaldehyde

O e 50 N f INt O N O F

55 To a dimethylsulfoxide (58.5 mL) solution of 2-fluoro-6- H (4-(E)-2-nitro-vinyl)-benzyloxy)-pyridine (3.44 g, 12.5 mmol) described in Manufacturing Example 55-1-3 and ace To a toluene (41 mL) solution of 4-(6-fluoro-pyridin-2- tic acid (3.44 mL) was added sodium borohydride (757 mg, yloxymethyl)-benzonitrile (5.99 g, 26.2 mmol) described in 20 mmol) at room temperature while cooling appropriately. Manufacturing Example 55-1-1 was added diisobutyl alumi 60 This mixture was stirred for 4 hours at room temperature. This num hydride (1.01 M toluene solution, 39.3 mmol) under mixture was partitioned into ethyl acetate and water. The nitrogen atmosphere at -70° C. to -78°C. This mixture was organic layer was separated, washed with Saturated aqueous stirred for 2 hours at room temperature. This mixture was Sodium chloride, dried over anhydrous magnesium sulfate, partitioned into ethyl acetate and 20% aqueous Rochelle salt and filtered. The filtrate was concentrated under a reduced solution. After removal of insoluble matter by filtrating 65 pressure, and the residue was purified by silica gel column through a Celite bed, the filtrate was partitioned. The organic chromatography (heptane:ethyl acetate=10: 1-4:1) to obtain layer was separated, washed with Saturated aqueous sodium the title compound (1.6 g., 46%). US 8,158,657 B2 189 190 H-NMR Spectrum (CDC1) & (ppm): 3.31-3.35 (2H, m), was stirred for 4 hours at 50° C. Water was added to the 4.60–4.63 (2H, m), 5.31 (2H, s), 6.48-6.50 (1H, m), 6.64-6.67 reaction Solution at room temperature, which was then (1H, m), 7.22-7.24 (2H, m), 7.41-7.43 (2H, m), 7.63-7.69 extracted with ethyl acetate. The organic layer was washed (1H, m). with Saturated aqueous Sodium chloride and dried over anhy drous magnesium sulfate, and the solvent was evaporated Manufacturing Example 55-1-5 under a reduced pressure. The residue was purified by NH silica gel column chromatography (heptane:ethyl acetate-4: (4-(6-Fluoro-pyridin-2-yloxymethyl)-phenyl)aceto 1-2: 1) to obtain the title compound (214 mg, 21%). hydroximoyl chloride H-NMR Spectrum (CDC1) & (ppm): 4.08 (2H, s), 5.08 10 (2H, s), 5.54 (2H, brs), 6.27 (1H, s), 6.71-6.74 (1H, m), 7.13-7.16 (1H, m), 7.31-7.39 (5H, m), 7.71-7.73 (1H, m), 8.11-8.14 (2H, m). e The starting material. (4-(5-fluoro-pyridin-2-yloxym O-N f ethyl)-phenyl)acetohydroximoyl chloride, was synthesized N as follows. F CIN Manufacturing Example 56-1-1 I HO 4-(5-Fluoro-pyridin-2-yloxymethyl)-benzonitrile

To a methanol (20 mL) solution of 2-fluoro-6-(4-(2-nitro ethyl)-benzyloxy)-pyridine (1.6 g., 5.79 mmol) described in Manufacturing Example 55-1-4 was added lithium methox ide (449 mg, 11.6 mmol). This mixture was stirred for 1 hour 25 at room temperature. The mixture was concentrated under a N2 N reduced pressure, water in the residue was azeotropically distilled with toluene, and this residue was diluted with meth To an N,N-dimethylformamide (50 mL) solution of ylene chloride (24 mL) and tetrahydrofuran (12 mL). This 2-bromo-5-fluoropyridine (5 g, 28.4 mmol) and 4-(hy was cooled to -78°C., and titanium (IV) tetrachloride (2.04 30 mL, 18.5 mmol) was added dropwise into the Suspension. droxymethyl)-benzonitrile (5.67 g. 42.4 mmol) was added This mixture was stirred for 2 hours at room temperature. This sodium hydride (1.7g, 42.4 mmol. 60% in oil) at room tem mixture was cooled to -78° C., and partitioned into ethyl perature. The mixture was stirred for 3 hours at 70° C. The acetate and ice water. The organic layer was separated, mixture was partitioned into ethyl acetate and water. The washed with Saturated aqueous Sodium chloride, dried over 35 organic layer was separated, washed with water, dried over anhydrous magnesium Sulfate, and filtered. The filtrate was anhydrous magnesium Sulfate, and filtered. The filtrate was concentrated under a reduced pressure, and the residue was concentrated under a reduced pressure, and the residue was triturated in ethyl acetate. The solid was collected and dried purified by NH-silica gel column chromatography (heptane: under reduced pressure to obtain the title compound (1.36 g). ethyl acetate-4:1-2: 1-1:1-ethyl acetate) to obtain the title This compound was used in the following reaction without 40 compound (5.5 g. 85%). further purification. 'H-NMR Spectrum (CDC1) & (ppm): 5.15 (2H, s), 7.14 7.17 (1H, m), 7.39-7.41 (1H, m), 7.53-7.55 (2H, m), 7.70 Example 56 7.72 (2H, m), 8.12-8.13 (1H, m). Manufacturing Example 56-1-2 3-(3-4-(5-Fluoro-pyridin-2-yloxymethyl)-benzyl)- 45 isoxazol-5-yl)-pyridin-2-ylamine 4-(5-Fluoro-pyridin-2-yloxymethyl)benzaldehyde

50

55 H To a toluene solution (37 mL) of 4-(5-fluoro-pyridin-2- yloxymethyl)-benzonitrile (5.5 g. 24.1 mmol) described in Manufacturing Example 56-1-1 was added diisobutyl alumi 60 num hydride (35.8 mL, 1.01 M toluene solution, 36.2 mmol) under nitrogen atmosphere at -70° C. to -78°C. This mixture To a tetrahydrofuran (5 mL) solution of ((4-(5-fluoro-py was stirred for 3 hours at room temperature. This mixture was ridin-2-yloxymethyl)-phenyl)acetohydroximoyl chloride partitioned into ethyl acetate and 20% aqueous Rochelle salt (800 mg, 2.72 mmol) described in Manufacturing example solution. After removal of insoluble matter by filtrating 56-1-5 and 3-ethynyl-pyridin-2-ylamine (200 mg, 1.69 65 through a Celite pad, the filtrate was partitioned. The organic mmol) described in Manufacturing Example 1-2-3 was added layer was separated, washed with Saturated aqueous sodium triethylamine (948 uL, 6.8 mmol) at room temperature, which chloride, dried over anhydrous magnesium sulfate, and fil