TWI415606B - Inhibitors of histone deacetylase - Google Patents
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本發明係關於組蛋白去乙醯基酶之抑制作用。更特定言之，本發明係關於抑制組蛋白去乙醯基酶酶活性的化合物及方法。The present invention relates to the inhibition of histone deacetylase. More specifically, the present invention relates to compounds and methods for inhibiting histone deacetylase enzymatic activity.
在真核細胞中，細胞核DNA與組蛋白結合形成稱作染色質之緊密複合物。組蛋白構成在真核物種中一般高度守恆的基礎蛋白質家族。稱為H2A、H2B、H3及H4的核心組蛋白結合形成蛋白核。DNA纏繞在該蛋白核上，同時組蛋白之鹼性胺基酸與帶負電荷的DNA磷酸基相互反應。將近146個DNA鹼基對環繞一組蛋白核組成一核小體粒子，即染色質之重複結構基元。In eukaryotic cells, nuclear DNA binds to histones to form a tight complex called chromatin. Histones constitute a family of basal proteins that are generally highly conserved in eukaryotic species. Core histones called H2A, H2B, H3 and H4 bind to form a protein core. DNA entangles on the nucleus, while the basic amino acid of the histone interacts with the negatively charged DNA phosphate. Nearly 146 DNA base pairs surround a group of protein cores to form a nucleosome particle, a repeating structural motif of chromatin.
Csordas,Biochem.J.,286：23－38(1990)教示組蛋白易受N－末端離胺酸殘基之ε－胺基轉譯後乙醯化作用，即一種由組蛋白乙醯基轉移酶催化之反應(HAT1)。乙醯化作用中和離胺酸側鏈之正電荷且被認為對染色質結構有影響。實際上，Taunton等人，Science,272：408－411(1996)教示藉由組蛋白超乙醯化作用放大轉錄因子至染色質模板的通路。Taunton等人還教示在基因組之轉錄沉默區中已發現富集未經乙醯化之組蛋白H4。Csordas, Biochem. J., 286: 23-38 (1990) teaches that histones are susceptible to ε-amino translation of N-terminal amino acid residues, followed by acetylation, a histone acetyltransferase Catalytic reaction (HAT1). The acetamylation neutralizes the positive charge of the amine acid side chain and is believed to have an effect on the chromatin structure. In fact, Taunton et al., Science, 272: 408-411 (1996) teach the amplification of transcription factors to chromatin template pathways by histone hyperacetylation. Taunton et al. also teach that enriched histone H4 has not been found in the transcriptional silence region of the genome.
組蛋白乙醯化作用係可逆修飾，且由稱為組蛋白去乙醯基酶(HDAC)之酶家族催化去乙醯作用。編碼具有HDAC活性之蛋白之基因序列的分子選殖已確立一組離散HDAC酶重整異構體之存在。Grozinger等人，Proc.Natl.Acad.Sci.USA,96：4868－4873(1999)教示HDAC分為兩類，第一類藉由酵母Rpd3－類蛋白質表示，且第二類藉由酵母Hdal－類蛋白質表示。Grozinger等人亦教示人類HDAC1、HDAC2及HDAC3蛋白質係第一類HDAC成員，且揭示命名為HDAC4、HDAC5及HDAC6之新蛋白質係第二類HDAC成員。Kao等人，Genes & Dev.,14：55－66(2000)揭示第二類HDAC之一新成員HDAC7。Van den Wyngaert,FEBS,478：77－83(2000)揭示第一類HDAC之一新成員HDAC8。Zhou.X.等人，Proc.Natl.Acad.Sci.U.S.A.98(19),10572－10577(2001)揭示HDAC9之選殖及表徵。Kao.H.Y.等人，J.Biol.chem.277(1),187－193(2002)揭示哺乳動物HDAC10之分離及表徵。Gao L.等人，JBio1 chem.277(28)：25748－55(2002)揭示HDAC11之選殖及功能表徵。Histone acetylation is reversibly modified and catalyzed by a family of enzymes called histone deacetylase (HDAC). Molecular colonization of a gene sequence encoding a protein having HDAC activity has established the existence of a discrete set of HDAC enzyme reformers. Grozinger et al., Proc. Natl. Acad. Sci. USA, 96: 4868-4873 (1999) teach that HDACs fall into two categories, the first being represented by the yeast Rpd3-like protein and the second being by the yeast Hdal- Protein-like representation. Grozinger et al. also teach the first class of HDAC members of the human HDAC1, HDAC2, and HDAC3 protein families, and reveal new protein-based HDAC members named HDAC4, HDAC5, and HDAC6. Kao et al, Genes & Dev., 14: 55-66 (2000) reveal a new member of the second class of HDAC, HDAC7. Van den Wyngaert, FEBS, 478: 77-83 (2000) reveals a new member of the first class of HDAC, HDAC8. Zhou.X. et al., Proc. Natl. Acad. Sci. U.S.A. 98(19), 10572-10577 (2001) discloses the selection and characterization of HDAC9. Kao. H. Y. et al., J. Biol. Chem. 277(1), 187-193 (2002) discloses the isolation and characterization of mammalian HDAC10. Gao L. et al, J Biol chem. 277 (28): 25748-55 (2002) reveal the selection and functional characterization of HDAC11.
Richon等人，Proc.Natl.Acad.Sci.USA,95：3003－3007(1998)揭示HDAC活性受到自吸水鏈黴菌(Streptomyces hygroscopicus)分離之天然產物曲古柳菌素A(trichostatin A)(TSA)抑制，及藉由合成化合物辛二醯苯胺異羥肟酸(suberoylanilide hydroxamic acid)(SAHA)抑制。Yoshida及Beppu,Exper.cell Res.,177：122－131(1988)教示TSA引起大鼠纖維母細胞在細胞週期在G1 及G2 階段停滯，說明HDAC涉及細胞循環調節中。實際上，Finnin等人，Nature,401：188－193(1999)教示TSA及SAHA抑制細胞生長，誘發末端分化且預防小鼠腫瘤形成。Suzuki等人，美國專利第6,174,905號、EP 0847992、JP 258863/96及日本專利申請案第10138957號揭示引起細胞分化並抑制HDAC之苯甲醯胺衍生物。Delorme等人之WO01/38322及PCT IB01/00683揭示充當HDAC抑制劑的其它化合物。Richon et al., Proc. Natl. Acad. Sci. USA, 95: 3003-3007 (1998) reveals that the HDAC activity is trichostatin A (TSA), a natural product isolated from Streptomyces hygroscopicus. Inhibition, and inhibition by the synthetic compound suberoylanilide hydroxamic acid (SAHA). Yoshida and Beppu, Exper. Cell Res., 177: 122-131 (1988) teach that TSA causes rat fibroblasts to arrest at the G 1 and G 2 phases of the cell cycle, indicating that HDAC is involved in cell cycle regulation. In fact, Finnin et al, Nature, 401: 188-193 (1999) teach that TSA and SAHA inhibit cell growth, induce terminal differentiation and prevent tumor formation in mice. Suzuki et al., U.S. Patent No. 6,174,905, EP 0 847 992, JP 258 863/96, and Japanese Patent Application No. 10 138 957 disclose benzamine derivatives which cause cell differentiation and inhibit HDAC. Other compounds that act as HDAC inhibitors are disclosed in WO 01/38322 to Delorme et al. and PCT IB01/00683.
該等發現表明抑制HDAC活性代表一種干預細胞週期調節之新穎途徑，且該等HDAC抑制劑具有治療細胞增殖性疾病或病症之巨大治療潛力。至今，此項技術中仍幾乎沒有組蛋白去乙醯基酶抑制劑。因此需要識別其它HDAC抑制劑及識別有效HDAC抑制活性所需要之結構特徵。These findings indicate that inhibition of HDAC activity represents a novel approach to intervention in cell cycle regulation, and that such HDAC inhibitors have enormous therapeutic potential for treating cell proliferative diseases or conditions. To date, there are still few histone deacetylase inhibitors in this technology. It is therefore necessary to identify other HDAC inhibitors and the structural features required to recognize potent HDAC inhibitory activity.
本發明提供用於治療細胞增殖性疾病之化合物及方法。The invention provides compounds and methods for treating cell proliferative disorders.
本發明提供組蛋白去乙醯基酶酶活性之新抑制劑。The present invention provides novel inhibitors of histone deacetylase enzymatic activity.
本發明之第一態樣提供可用作組蛋白去乙醯基酶抑制劑之化合物。A first aspect of the invention provides a compound useful as a histone deacetylase inhibitor.
本發明之第二態樣提供一種組合物，其包含本發明之組蛋白去乙醯基酶抑制劑及醫藥學上可接受之載劑、賦形劑或稀釋劑。A second aspect of the invention provides a composition comprising a histone deacetylase inhibitor of the invention and a pharmaceutically acceptable carrier, excipient or diluent.
本發明之第三態樣提供一種抑制一細胞中組蛋白去乙醯基酶之方法，其包含使一希望抑制組蛋白去乙醯基酶之細胞與本發明之組蛋白去乙醯基酶抑制劑接觸。A third aspect of the present invention provides a method for inhibiting histone deacetylase in a cell, comprising: inhibiting a histone deacetylase-depressing cell and a histone deacetylase inhibitor of the present invention Agent contact.
本發明之第四態樣提供一種用於治療細胞增殖性疾病之方法。A fourth aspect of the invention provides a method for treating a cell proliferative disorder.
前述者僅概述本發明之特定態樣而實質上不欲進行限制。該等態樣及其它態樣及實施例更充分描述於下文。The foregoing is merely illustrative of specific aspects of the invention. These and other aspects and embodiments are more fully described below.
本發明提供抑制組蛋白去乙醯基酶酶活性之化合物及方法。本發明亦提供用於治療細胞增殖性疾病及病症之組合物及方法。本文引用之專利及科學文獻建立熟習此項技術者可獲得之知識。本文列舉之經頒予專利、申請案及參考文獻以引用之方式併入本文中，該引用程度如同已特定及個別地將每一者以引用之方式併入一般。在發生矛盾之狀況中將採用本發明之揭示內容。The present invention provides compounds and methods for inhibiting histone deacetylase enzymatic activity. The invention also provides compositions and methods for treating cell proliferative diseases and disorders. The patents and scientific literature cited herein establish knowledge available to those skilled in the art. The patents, applications, and references cited herein are hereby incorporated by reference in their entirety in their entirety in the extent the the the the the The disclosure of the present invention will be employed in the event of a conflict.
為本發明之目的將採用以下定義(除非另有明確說明)：本文所用術語"組蛋白去乙醯基酶"及"HDAC"欲指自組蛋白N－末端之離胺酸殘基之ε－胺基去除乙醯基的酶家族中任何一員。除非上下文另有說明，術語"組蛋白"欲指包括來自任何種類之H1、H2A、H2B、H3、H4及H5之任何組蛋白蛋白質。較佳之組蛋白去乙醯基酶包括I類及II類酶。較佳組蛋白去乙醯基酶為人類HDAC，其包括但不限於：HDAC－1、HDAC－2、HDAC－3、HDAC－4、HDAC－5、HDAC－6、HDAC－7、HDAC－8、HDAC－9、HDAC－10及HDAC－11。一些其它較佳實施例中，該組蛋白去乙醯基酶源自原生動物或真菌源。The following definitions will be used for the purposes of the present invention (unless otherwise explicitly stated): The terms "histone deacetylase" and "HDAC" as used herein are intended to mean the epsilon of the amino acid residue from the N-terminus of the histone. Any one of the family of enzymes that removes the thiol group from the amine group. Unless the context indicates otherwise, the term "histone" is intended to include any histone protein from any of the classes H1, H2A, H2B, H3, H4 and H5. Preferred histone deacetylases include class I and class II enzymes. Preferred histone deacetylases are human HDACs including, but not limited to, HDAC-1, HDAC-2, HDAC-3, HDAC-4, HDAC-5, HDAC-6, HDAC-7, HDAC-8 , HDAC-9, HDAC-10 and HDAC-11. In some other preferred embodiments, the histone deacetylase is derived from a protozoan or fungal source.
術語"組蛋白去乙醯基酶抑制劑"用於識別具有本文定義之結構的化合物，其能夠與組蛋白去乙醯基酶相互作用且抑制其酶活性。"抑制組蛋白去乙醯基酶酶活性"意謂降低組蛋白去乙醯基酶自組蛋白移除乙醯基的能力。一些較佳實施例中，組蛋白去乙醯基酶活性降低率為至少約50%，更佳至少約75%，且仍更佳至少約90%。在其它較佳實施例中，組蛋白去乙醯基酶活性減少至少95%且更佳減少至少99%。The term "histone deacetylase inhibitor" is used to identify a compound having a structure as defined herein which is capable of interacting with histone deacetylase and inhibiting its enzymatic activity. "Inhibition of histone deacetylase enzymatic activity" means reducing the ability of histone deacetylase to remove acetamyl groups from histones. In some preferred embodiments, the rate of histone deacetylase activity reduction is at least about 50%, more preferably at least about 75%, and still more preferably at least about 90%. In other preferred embodiments, the histone deacetylase activity is reduced by at least 95% and more preferably by at least 99%.
該抑制較佳為特異性的，即組蛋白去乙醯基酶抑制劑降低組蛋白去乙醯基酶自組蛋白移除乙醯基之能力，該抑制劑濃度低於產生其它無關生物效應所需之抑制劑濃度。組蛋白去乙醯基酶抑制活性所需之抑制劑濃度較佳比產生無關生物效應所需之濃度至少低2倍，更佳至少低5倍，甚至更佳至少低10倍，且最佳至少低20倍。Preferably, the inhibition is specific, that is, the histone deacetylase inhibitor reduces the ability of the histone deacetylase to remove the acetyl group from the histone, which is lower than other unrelated biological effects. The concentration of inhibitor required. The concentration of the inhibitor required for the histone deacetylase inhibitory activity is preferably at least 2 times lower than the concentration required to produce an unrelated biological effect, more preferably at least 5 times lower, even more preferably at least 10 times lower, and optimally at least 20 times lower.
為簡單起見，全文主要將化學部分定義及稱作單價化學部分(例如烷基、芳基等)。儘管如此，該等術語亦用於表達在熟習此項技術者所清楚的適當結構情況下之相應多價部分。舉例而言，雖然"烷基"部分一般係指單價基(例如CH3 －CH2 －)，但在特定情況下二價連接部分可為"烷基"，該狀況中熟習此項技術者應瞭解該烷基為等效於術語"伸烷基"之二價基(例如－CH2 －CH2 －)。(同樣地，在需要二價部分且將其稱作"芳基"的情況中，熟習此項技術者應瞭解術語"芳基"係指相應二價部分，即伸芳基。)應瞭解所有原子具有成鍵標準價數(即，碳為4、N為3、O為2、硫視其氧化態而定為2、4或6)。有時部分可經定義，例如(A)a －B－，其中a為0或1。在該等情況下，當a為0時該部分為B－且當a為1時該部分為A－B－。For the sake of simplicity, the chemical part is mainly defined and referred to as a monovalent chemical moiety (eg, alkyl, aryl, etc.). Nonetheless, the terms are used to denote corresponding multivalent portions of the appropriate structure as would be apparent to those skilled in the art. For example, although the "alkyl" moiety generally refers to a monovalent group (e.g., CH 3 -CH 2 -), but the divalent linking moiety may under certain circumstances as "alkyl", the condition skilled in the art should It is understood that the alkyl group is equivalent to the divalent group of the term "alkylene" (for example -CH 2 -CH 2 -). (Again, in the case where a divalent moiety is required and referred to as "aryl", those skilled in the art should understand that the term "aryl" refers to the corresponding divalent moiety, ie, an extended aryl group.) The atom has a bond valence (ie, carbon is 4, N is 3, O is 2, and sulfur is 2, 4, or 6 depending on its oxidation state). Sometimes parts can be defined, such as (A) a - B-, where a is 0 or 1. In such cases, when a is 0, the portion is B- and when a is 1, the portion is A-B-.
術語"烴基"係指各如本文所定義之直鏈、支鏈或環狀烷基、烯基或炔基。"C0 "烴基係指共價鍵。因此"C0 －C3 －烴基"包括共價鍵、甲基、乙基、乙烯基、乙炔基、丙基、丙烯基、丙炔基及環丙基。The term "hydrocarbyl" refers to a straight, branched or cyclic alkyl, alkenyl or alkynyl group, each as defined herein. "C 0 "hydrocarbyl refers to a covalent bond. Thus "C 0 -C 3 -hydrocarbyl" includes covalent bonds, methyl, ethyl, vinyl, ethynyl, propyl, propenyl, propynyl and cyclopropyl.
本文採用之術語"烷基"係指具有自1至12個碳原子、較佳1－8個碳原子、更佳1－6個碳原子之直鏈及支鏈脂族基，該等脂族基視情況經一、二或三個取代基取代。較佳烷基包括(不限於)甲基、乙基、丙基、異丙基、丁基、異丁基、第二丁基、第三丁基、戊基及己基。"C0 "烷基(如"C0 －C3 －烷基")為共價鍵(類似於"C0 "烴基)。The term "alkyl" as used herein means a straight-chain or branched aliphatic radical having from 1 to 12 carbon atoms, preferably from 1 to 8 carbon atoms, more preferably from 1 to 6 carbon atoms, such aliphatic groups. The base-view condition is replaced by one, two or three substituents. Preferred alkyl groups include, without limitation, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, second butyl, tert-butyl, pentyl and hexyl. A "C 0 "alkyl group (such as "C 0 -C 3 -alkyl") is a covalent bond (similar to a "C 0 "hydrocarbyl group).
本文所用術語"烯基"意謂具有一或多個碳－碳雙鍵之不飽和直鏈或支鏈脂族基，該等脂族基具有2至12個原子、較佳2－8個碳原子、更佳2－6個碳原子且視情況經一、二或三個取代基取代。較佳烯基包括(不限於)乙烯基、丙烯基、丁烯基、戊烯基及己烯基。The term "alkenyl" as used herein, means an unsaturated straight or branched aliphatic radical having one or more carbon-carbon double bonds having from 2 to 12 atoms, preferably from 2 to 8 carbons. The atom, more preferably 2 to 6 carbon atoms, is optionally substituted with one, two or three substituents. Preferred alkenyl groups include, without limitation, ethenyl, propenyl, butenyl, pentenyl, and hexenyl.
本文所用術語"炔基"意謂具有一或多個碳－碳三鍵之不飽和直鏈或支鏈脂族基，該等脂族基具有2至12個碳原子、較佳2至8個碳原子、更佳2－6個碳原子且視情況經一、二或三個取代基取代。較佳之炔基包括(不限於)乙炔基、丙炔基、丁炔基、戊炔基及己炔基。The term "alkynyl" as used herein, means an unsaturated straight or branched aliphatic radical having one or more carbon-carbon triple bonds, the aliphatic groups having from 2 to 12 carbon atoms, preferably from 2 to 8 The carbon atom, more preferably 2 to 6 carbon atoms, is optionally substituted with one, two or three substituents. Preferred alkynyl groups include, without limitation, ethynyl, propynyl, butynyl, pentynyl and hexynyl.
"伸烷基"、"伸烯基"或"伸炔基"為如上文定義之烷基、烯基或炔基，其係位於兩個其它化學基團之間且用以連接它們。較佳之伸烷基包括(不限於)：亞甲基、伸乙基、伸丙基及伸丁基。較佳之伸烯基包括(不限於)：伸乙烯基、伸丙烯基及伸丁烯基。較佳之伸炔基包括(不限於)：伸乙炔基、伸丙炔基及伸丁炔基。"Alkyl", "alkenyl" or "alkenyl" are alkyl, alkenyl or alkynyl groups as defined above which are positioned between two other chemical groups and are used to attach them. Preferred alkylene groups include, without limitation, methylene, ethyl, propyl and butyl. Preferred alkenyl groups include, without limitation, vinyl, propylene, and butenyl groups. Preferred alkynyl groups include, without limitation, ethynyl, extended propynyl, and butynyl groups.
本文所採用之術語"環烷基"包括具有3至12個碳原子、較佳3至8個碳原子、更佳3至6個碳原子之飽和及部分不飽和之環烴基，其中該環烷基另外視情況經取代。較佳之環烷基包括(不限於)：環丙基、環丁基、環戊基、環戊烯基、環己基、環己烯基、環庚基及環辛基。The term "cycloalkyl" as used herein, includes saturated and partially unsaturated cyclic hydrocarbon groups having from 3 to 12 carbon atoms, preferably from 3 to 8 carbon atoms, more preferably from 3 to 6 carbon atoms, wherein the cycloalkane The base is replaced by another situation. Preferred cycloalkyl groups include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl and cyclooctyl.
術語"雜烷基"係指如上文定義之烷基，其中鏈中一或多個碳原子經選自由O、S及N所組成之群的雜原子取代。The term "heteroalkyl" refers to an alkyl group as defined above wherein one or more carbon atoms in the chain are substituted with a heteroatom selected from the group consisting of O, S and N.
"芳基"係為包含一至三個芳族環之C6 －C1 4 芳族部分，該芳族部分視情況經取代。該芳基較佳為C6 －C1 0 芳基。較佳之芳基包括(不限於)苯基、萘基、蒽基及茀基。"芳烷基(aralkyl)"或"芳基烷基(arylalkyl)"包含共價連接至烷基之芳基，其任一者可獨立視情況經取代或未經取代。該芳烷基較佳為(C1 －C6 )烷(C6 －C1 0 )芳基，包括(不限於)苄基、苯乙基及萘甲基。"Aryl" system comprising one to three aromatic rings C 6 -C 1 4 aromatic moiety, the aromatic moiety optionally substituted. The aryl group is preferably a C 6 -C 1 0 aryl group. Preferred aryl groups include, without limitation, phenyl, naphthyl, anthryl and fluorenyl. "Aralkyl" or "arylalkyl" includes an aryl group covalently bonded to an alkyl group, either of which may be substituted or unsubstituted, as appropriate. The aralkyl group is preferably a (C 1 -C 6 )alkane (C 6 -C 1 0 )aryl group including, without limitation, a benzyl group, a phenethyl group, and a naphthylmethyl group.
"雜環基"("heterocyclic group"或"heterocyclyl")視情況經取代自約3至約14個原子之非芳族單、二或三環結構，其中一或多個原子係選自由N、O及S所組成之群。雙環雜環中之一個環或三環雜環中之兩個環可為芳族，如茚滿及9,10－二氫蒽。該雜環基視情況經酮基或上述取代基中之一者在碳上取代。該雜環基亦可獨立經烷基、芳基、芳烷基、烷羰基、烷基磺醯基、芳羰基、芳基磺醯基、烷氧羰基、芳烷氧羰基在氮上取代，或經酮基或低碳烷基在硫上取代。較佳之雜環基包括(不限於)：環氧基、氮丙啶基、四氫呋喃基、吡咯啶基、哌啶基、六氫吡嗪基、噻唑烷基、噁唑啶基、噁唑啶酮基及嗎啉基。在特定較佳實施例中，將雜環基稠合至芳基、雜芳基或環烷基。該等稠合雜環之實例包括(不限於)四氫喹啉及二氫苯幷呋喃。自該術語之範疇特定排除其中環O或S原子相鄰於另一O或S原子之化合物。"Heterocyclic group" ("heterocyclic group" or "heterocyclyl") is optionally substituted with a non-aromatic mono-, bi- or tricyclic structure of from about 3 to about 14 atoms, wherein one or more of the atoms are selected from N, A group of O and S. Two of the bicyclic or tricyclic heterocyclic rings may be aromatic, such as indane and 9,10-dihydroanthracene. The heterocyclic group is optionally substituted on the carbon via a ketone group or one of the above substituents. The heterocyclic group may also be independently substituted on the nitrogen via an alkyl group, an aryl group, an arylalkyl group, an alkylcarbonyl group, an alkylsulfonyl group, an arylcarbonyl group, an arylsulfonyl group, an alkoxycarbonyl group, an aralkoxycarbonyl group, or Substitution with a keto group or a lower alkyl group on sulfur. Preferred heterocyclic groups include, without limitation: epoxy, aziridine, tetrahydrofuranyl, pyrrolidinyl, piperidinyl, hexahydropyrazinyl, thiazolidinyl, oxazolidinyl, oxazolidinone And morpholinyl. In a particularly preferred embodiment, the heterocyclic group is fused to an aryl, heteroaryl or cycloalkyl group. Examples of such fused heterocycles include, without limitation, tetrahydroquinoline and dihydrobenzoquinone. Compounds in which the ring O or S atom is adjacent to another O or S atom are specifically excluded from the scope of the term.
本文所用術語"雜芳基"係指視情況經取代之基團，其具有5至14個環原子、較佳5、6、9或10個環原子；具有6、10或14個在一環狀排列共享之π電子；且具有除碳原子之外選自由N、O及S所組成之群的一或多個雜原子。舉例而言，雜芳基可為嘧啶基、吡啶基、苯幷咪唑基、噻吩基、苯幷噻唑基、苯幷呋喃基及吲哚啉基。較佳之雜芳基包括(不限於)：噻吩基、苯幷噻吩基、呋喃基、苯幷呋喃基、二苯幷呋喃基、吡咯基、咪唑基、吡唑基、吡啶基、吡嗪基、嘧啶基、吲哚基、喹啉基、異喹啉基、喹噁啉基、四唑基、噁唑基、噻唑基、***基及異噁唑基。The term "heteroaryl" as used herein, refers to a optionally substituted group having from 5 to 14 ring atoms, preferably 5, 6, 9 or 10 ring atoms; having 6, 10 or 14 rings in one ring. The shared π electrons are arranged in an array; and have one or more hetero atoms selected from the group consisting of N, O and S in addition to carbon atoms. For example, a heteroaryl group can be a pyrimidinyl group, a pyridyl group, a benzimidazolyl group, a thienyl group, a benzothiazolyl group, a benzofuranyl group, and a porphyrin group. Preferred heteroaryl groups include, but are not limited to, thienyl, benzothiophenyl, furyl, benzofuranyl, diphenylfuranyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, Pyrimidinyl, fluorenyl, quinolyl, isoquinolyl, quinoxalinyl, tetrazolyl, oxazolyl, thiazolyl, triazolyl and isoxazolyl.
"雜芳烷基"("heteroaralkyl"或"heteroarylalkyl")包含共價連接至烷基之雜芳基，其獨立視情況經取代或未經取代。較佳之雜烷基包含C1 －C6 烷基及具有5、6、9或10個環原子之雜芳基。自該術語之範疇特定排除具有相鄰環O及/或S原子之化合物。較佳之雜芳烷基實例包括：吡啶基甲基、吡啶基乙基、吡咯基甲基、吡咯基乙基、咪唑基甲基、咪唑基乙基、噻唑基甲基及噻唑基乙基。"Heteroaralkyl" or "heteroarylalkyl" includes a heteroaryl group covalently bonded to an alkyl group, which is optionally substituted or unsubstituted, as appropriate. Preferred heteroalkyl groups include C 1 -C 6 alkyl groups and heteroaryl groups having 5, 6, 9 or 10 ring atoms. Compounds having adjacent ring O and/or S atoms are specifically excluded from the scope of the term. Examples of preferred heteroaralkyl groups include pyridylmethyl, pyridylethyl, pyrrolylmethyl, pyrrolylethyl, imidazolylmethyl, imidazolylethyl, thiazolylmethyl and thiazolylethyl.
"伸芳基"、"伸雜芳基"或"伸雜環基"為如上文定義之芳基、雜芳基或雜環基，其係位於中間且用於連接兩個其它化學基團。"Exoaryl", "heteroaryl" or "heterocyclyl" is an aryl, heteroaryl or heterocyclyl radical as defined above which is intermediate and is used to link two other chemical groups.
較佳之雜環基及雜芳基包括(但不限於)：吖啶基、氮基、苯幷咪唑基、苯幷呋喃基、苯幷硫代呋喃基、苯幷噻吩基、苯幷噁唑基、苯幷噻唑基、苯幷***基、苯幷四唑基、苯幷異噁唑基、苯幷異噻唑基、苯幷咪唑啉基、咔唑基、4aH－咔唑基、咔啉基、基、烯基、噌啉基、十氫喹啉基、2H,6H－1,5,2－二噻嗪基、二氫呋喃幷[2,3－b]四氫呋喃、呋喃基、呋吖基、咪唑啶基、咪唑啉基、咪唑基、1H－吲唑基、伸吲哚基(indolenyl)、吲哚啉基,吲嗪基、吲哚基、3H－吲哚基、異苯幷呋喃基、異基、異吲唑基、異吲哚啉基、異吲哚基、異喹啉基、異噻唑基、異噁唑基、亞甲基二氧苯基、嗎啉基、啶基、八氫異喹啉基、噁二唑基、1,2,3－噁二唑基、1,2,4－噁二唑基、1,2,5－噁二唑基、1,3,4－噁二唑基、噁唑啶基、噁唑基、噁唑啶基、嘧啶基、啡啶基、啡啉基、啡嗪基、啡噻嗪基、氧硫雜蒽基、啡噁嗪基、呔嗪基、六氫吡嗪基、哌啶基、哌啶酮基、4－哌啶酮基、向日葵基、喋啶基、嘌呤基、哌喃基、吡嗪基、吡唑啶基、吡唑啉基、吡唑基、嗒嗪基、吡啶幷噁唑、吡啶幷咪唑、吡啶幷噻唑、吡啶基(pyridinyl)、吡啶基(pyridyl)、嘧啶基、吡咯啶基、吡咯啉基、2H－吡咯基、吡咯基、喹唑啉基、喹啉基、4H－喹嗪基、喹噁啉基、啶基、四氫呋喃基、四氫異喹啉基、四氫喹啉基、四唑基、6H－1,2,5－噻二嗪基、1,2,3－噻二唑基、1,2,4－噻二唑基、1,2,5－噻二唑基、1,3,4－噻二唑基、噻嗯基、噻唑基、噻吩基、噻吩幷噻唑基、噻吩幷噁唑基、噻吩幷咪唑基、噻吩基、三嗪基、1,2,3－***基、1,2,4－***基、1,2,5－***基、1,3,4－***基及基。Preferred heterocyclic and heteroaryl groups include, but are not limited to, acridinyl, nitrogen Base, benzoimidazolyl, benzofuranyl, phenylsulfonylfuranyl, benzoquinone, benzoxazolyl, benzothiazolyl, benzotriazolyl, benzodiazolyl, benzoquinone Oxazolyl, benzoquinoneisothiazolyl, phenylimidazolidinyl, carbazolyl, 4aH-carbazolyl, porphyrinyl, base, Alkenyl, porphyrin, decahydroquinolyl, 2H, 6H-1,5,2-dithiazinyl, dihydrofuranium [2,3-b]tetrahydrofuran, furyl, furazyl, imidazolidinium , imidazolinyl, imidazolyl, 1H-carbazolyl, indolenyl, porphyrinyl, pyridazinyl, fluorenyl, 3H-fluorenyl, isophthalide, iso Base, isoxazolyl, isoindolyl, isodecyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylene dioxyphenyl, morpholinyl, Pyridyl, octahydroisoquinolyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1, 3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, pyrimidinyl, morphinyl, morpholinyl, cyanozinyl, phenothiazine, oxazinyl, morphine Oxazinyl, pyridazinyl, hexahydropyrazinyl, piperidinyl, piperidinone, 4-piperidinone, sunflower, acridinyl, fluorenyl, piperidyl, pyrazinyl, pyrazole Pyridyl, pyrazolinyl, pyrazolyl, pyrazinyl, pyridoxazole, pyridinium imidazole, pyridinium thiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrroline Base, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolyl, 4H-quinazinyl, quinoxalinyl, Pyridyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolyl, tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2 , 4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thienyl, thiazolyl, thienyl, thiophenethiazolyl, thiophenoxazolyl , thiophene imidazolyl, thienyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-tri Azolyl and base.
如本文所採用，當將一個部分(例如環烷基、烴基、芳基、雜芳基、雜環者、尿素等)描述為"視情況經取代"時其意謂該基團視情況具有一至四個、較佳一至三個、更佳一或兩個非氫取代基。合適之取代基包括(不限於)：鹵基、羥基、酮基(例如經酮基取代之環－CH－為－C(O)－)、硝基、鹵代烴基、烴基、芳基、芳烷基、烷氧基、鹵代烷氧基、芳氧基、雜芳氧基、胺基、醯胺基、烷基胺甲醯基、芳基胺甲醯基、胺基烷基、醯基、羧基、羥烷基、烷磺醯基、芳烴磺醯基、磺醯胺基、烷磺醯胺基、芳烴磺醯胺基、芳烷基磺醯胺基、烷羰基、醯氧基、氰基、烷硫基、脲基及脲烷基。其本身未經進一步取代(除非另有明確說明)之較佳取代基為：(a)鹵基、氰基、酮基、烷基、烷氧基、烷硫基、鹵代烷氧基、胺基烷基、胺基烷氧基、羧基、甲醯基、硝基、胺基、甲脒基、胺甲醯基、胍基、C3 －C7 雜環基、雜環基烷基、雜環羰基、羥烷基、烷氧基烷基，(b)C1 －C5 烷基或烯基或芳烷基亞胺基、胺甲醯基、胺基甲酸酯、疊氮基、甲醯胺基、巰基、羥基、羥烷基、烷芳基、芳烷基、C1 －C8 烷基、C1 －C8 烯基、C1 －C8 烷氧基、C1 －C8 烷氧羰基、芳氧羰基、C2 －C8 醯基、C2 －C8 醯胺基、C1 －C8 烷硫基、芳基烷硫基、芳硫基、雜芳硫基、C1 －C8 烷基亞磺醯基、芳烷基亞磺醯基、芳基亞磺醯基、C1 －C8 烷基磺醯基、芳烷基磺醯基、芳基磺醯基、C0 －C6 N－烷基胺甲醯基、C2 －C1 5 N,N－二烷基胺甲醯基、C3 －C7 環烷基、芳醯基、芳氧基、雜芳氧基、芳烷基醚、C3 －C7 雜環基烷基醚、芳基、稠合至環烷基或雜環或另一芳基環之芳基、C3 －C7 雜環、雜芳基、芳基胺甲醯基，或稠合或螺－稠合至環烷基、雜環基或芳基之該等環任一者，其中前述任何可再經取代者視情況又經一或多個列於上述(a)之部分取代；及(c)－(CH2 )s －NR3 0 R3 1 ，其中s係0(該狀況下氮直接結合經取代之部分)至6，且R3 0 及R3 1 係各自獨立為：氫、氰基、酮基、甲醯胺基、甲脒基、C1 －C8 羥烷基、C1 －C3 烷基芳基、芳基－C1 －C3 烷基、C1 －C8 烷基、C1 －C8 烯基、C1 －C8 烷氧基、C1 －C8 烷氧羰基、芳氧羰基、芳基－C1 －C3 烷氧羰基、C2 －C8 醯基、C1 －C8 烷基磺醯基、芳烷基磺醯基、芳基磺醯基、芳醯基、芳基、環烷基、雜環基或雜芳基，其中各前述者視情況又經一或多個列於上述(a)之部分取代；或R3 0 及R3 1 連同其所連接之氮形成各視情況經1至3個列於前述(a)之取代基所取代之雜環基或雜芳基。As used herein, when a moiety (eg, cycloalkyl, hydrocarbyl, aryl, heteroaryl, heterocycle, urea, etc.) is described as "optionally substituted", it means that the group has one to Four, preferably one to three, more preferably one or two non-hydrogen substituents. Suitable substituents include, without limitation: halo, hydroxy, keto (eg, keto substituted ring -CH- is -C(O)-), nitro, halohydrocarbyl, hydrocarbyl, aryl, aryl Alkyl, alkoxy, haloalkoxy, aryloxy, heteroaryloxy, amine, amidino, alkylamine, arylaminomethyl, arylalkyl, aryl, carboxy , hydroxyalkyl, alkanesulfonyl, arylsulfonyl, sulfonylamino, alkanesulfonylamino, arylsulfonylamino, aralkylsulfonylamino, alkylcarbonyl, decyloxy, cyano, Alkylthio, ureido and ureido. Preferred substituents which are not themselves further substituted (unless otherwise explicitly stated) are: (a) halo, cyano, keto, alkyl, alkoxy, alkylthio, haloalkoxy, aminyl Base, aminoalkoxy group, carboxyl group, formyl group, nitro group, amine group, formyl group, amine formyl group, fluorenyl group, C 3 -C 7 heterocyclic group, heterocyclic group alkyl group, heterocyclic carbonyl group , hydroxyalkyl, alkoxyalkyl, (b) C 1 -C 5 alkyl or alkenyl or aralkylimine, amine carbaryl, urethane, azide, formamide Base, mercapto, hydroxy, hydroxyalkyl, alkaryl, aralkyl, C 1 -C 8 alkyl, C 1 -C 8 alkenyl, C 1 -C 8 alkoxy, C 1 -C 8 alkoxy Carbonyl, aryloxycarbonyl, C 2 -C 8 fluorenyl, C 2 -C 8 decylamino, C 1 -C 8 alkylthio, arylalkylthio, arylthio, heteroarylthio, C 1 - C 8 alkylsulfinyl, arylalkylsulfinyl, arylsulfinyl, C 1 -C 8 alkylsulfonyl, aralkylsulfonyl, arylsulfonyl, C 0 -C 6 N- alkyl amine acyl, C 2 -C 1 5 N, N- dialkyl carbamoyl acyl, C 3 -C 7 cycloalkyl, aryl, acyl, aryloxy, heteroaryl Group, an aryl alkyl ether, C 3 -C 7 heterocyclyl alkyl ether, an aryl group, fused to a cycloalkyl or heterocyclic or another aryl ring, the aryl group, C 3 -C 7 heterocycle, a heteroaryl group, an arylamine carbenyl group, or a ring which is fused or spiro-fused to a cycloalkyl group, a heterocyclic group or an aryl group, wherein any of the foregoing re-substitutables may be One or more of the substitutions listed in (a) above; and (c)-(CH 2 ) s -NR 3 0 R 3 1 , wherein s is 0 (in this case, the nitrogen directly binds to the substituted moiety) to 6 And R 3 0 and R 3 1 are each independently: hydrogen, cyano, keto, formamidine, formamyl, C 1 -C 8 hydroxyalkyl, C 1 -C 3 alkylaryl, Aryl-C 1 -C 3 alkyl, C 1 -C 8 alkyl, C 1 -C 8 alkenyl, C 1 -C 8 alkoxy, C 1 -C 8 alkoxycarbonyl, aryloxycarbonyl, aryl -C 1 -C 3 alkoxycarbonyl, C 2 -C 8 fluorenyl, C 1 -C 8 alkylsulfonyl, arylalkylsulfonyl, arylsulfonyl, arylsulfonyl, aryl, cycloalkyl, aryl, heterocyclyl or heteroaryl, wherein each of the foregoing are optionally substituted with one or more further substituents listed in the above section (a); or R 3 0 and R 3 1 with its Forming each of the connecting nitrogen optionally substituted with 1 to 3 substituents listed in the preceding (a) substituents of the heterocyclyl or heteroaryl group.
另外，環部分上之取代基(即環烷基、雜環基、芳基、雜芳基)包括稠合至母環部分以形成雙環或三環稠合環系統之5－6員單環及9－14員雙環部分。舉例而言，視情況經取代之苯基包括但不限於如下基團：
環部分上之較佳取代基(即環烷基、雜環基、芳基、雜芳基)亦包括式－K1 －N(H)(R1 0 )之基團，其中K1 為化學鍵或C1 －C4 伸烷基；R1 0 係選自由Z'及－Ak2 －Z'所組成之群，其中Ak2 為C1 －C4 伸烷基；及Z'為環烷基、芳基、雜芳基或雜環基，其各視情況經取代且各視情況稠合至一或多個芳基或雜芳基環，或稠合至一或多個飽和或部分不飽和環烷基或雜環基環。Preferred substituents on the ring moiety (ie, cycloalkyl, heterocyclyl, aryl, heteroaryl) also include groups of the formula -K 1 -N(H)(R 1 0 ) wherein K 1 is a chemical bond Or a C 1 -C 4 alkylene group; R 10 0 is selected from the group consisting of Z′ and —Ak 2 —Z′, wherein Ak 2 is a C 1 -C 4 alkylene group; and Z′ is a cycloalkyl group. , aryl, heteroaryl or heterocyclyl, each optionally substituted and optionally fused to one or more aryl or heteroaryl rings, or fused to one or more saturated or partially unsaturated A cycloalkyl or heterocyclyl ring.
環狀部分(諸如芳基、雜芳基、環烷基、雜環基或稠合至一或多個芳基或雜芳基環或稠合至一或多個飽和或部分不飽和之環烷基或雜環基環的此等基團任一者)上尤其較佳之取代基包括1、2或3個獨立選自以下基團者：a)烷氧基、氰基、胺基、酮基、鹵代烷基、鹵基、烷基、R5 0 －C(O)－N(R3 2 )－、R5 0 －O－C(O)－N(R3 2 )－、R5 0 －NH－C(O)－N(R3 2 )－、R5 0 －NH－C(O)－O－、(R3 2 )(R3 3 )N－烷基－、(R3 2 )(R3 3 )N－烷基－O－、(R3 2 )(R3 3 )N－伸烯基－N(R3 2 )－、N(R3 2 )－芳基－N(R3 2 )－C(O)－芳基－烷基－N(R3 2 )－；其中R5 0 為環烷基、雜環基－C1 －C6 烷基－、R3 2 R3 3 N－烷基－或烷基；b)芳基－C0 －C6 烷基－、雜芳基－C0 －C6 烷基－、環烷基－C0 －C6 烷基－、雜環基－C0 －C6 烷基－、芳基－C0 －C6 烷基－N(R3 2 )－、芳基－C(O)－、雜芳基－C0 －C6 烷基－N(R3 2 )－、雜環基－C0 －C6 烷基－N(R3 2 )－、芳基－O－、雜芳基－O－、芳基－S－、雜芳基－S－、芳基－SO2 －、雜芳基－SO2 －、芳基－C(O)N(R3 2 )－、雜芳基－C(O)N(R3 2 )－、雜芳基－C(H)(SO2 －雜芳基)－N(R3 2 )－；其中R3 2 及R3 3 係獨立為H或C1 －C6 烷基，或R3 2 及R3 3 連同其所連接之N形成一雜環基或雜芳基；且其中[0045]－[0048]段描述之任一環進一步視情況經1、2或3個獨立選自以下基團者取代：烷基、烷氧基、硫代烷氧基、烷基－SO2 －、胺基、鹵基、氰基、鹵代烷基、羥烷基、烷氧烷氧烷基、COOH、烷醯基、鏈烷酸酯、NO2 、羥基、鹵代烷氧基、(R3 2 )(R3 3 )N－C0 －C6 烷基－、(R3 2 )(R3 3 )N－C0 －C6 烷基－O－、(R3 2 )(R3 3 )N－C(O)－、雜芳基、烷基－C(O)N(R3 2 )－、芳基－O－、(R3 2 )(R3 3 )N－SO2 －、芳基及(R3 2 )(R3 3 )N－烷基－C(O)N(R3 2 )－。a cyclic moiety (such as an aryl, heteroaryl, cycloalkyl, heterocyclyl or fused to one or more aryl or heteroaryl rings or fused to one or more saturated or partially unsaturated naphthenes) Particularly preferred substituents on any of these groups of a heterocyclic ring or a heterocyclyl ring include 1, 2 or 3 independently selected from the group consisting of: a) alkoxy, cyano, amine, keto , haloalkyl, halo, alkyl, R 5 0 -C(O)-N(R 3 2 )-, R 5 0 -O-C(O)-N(R 3 2 )-, R 5 0 - NH-C(O)-N(R 3 2 )-, R 5 0 -NH-C(O)-O-, (R 3 2 )(R 3 3 )N-alkyl-, (R 3 2 ) (R 3 3 )N-alkyl-O-, (R 3 2 )(R 3 3 )N-alkenyl-N(R 3 2 )-, N(R 3 2 )-aryl-N(R 3 2 )-C(O)-aryl-alkyl-N(R 3 2 )-; wherein R 5 0 is cycloalkyl, heterocyclyl-C 1 -C 6 alkyl-, R 3 2 R 3 3 N-alkyl- or alkyl; b) aryl-C 0 -C 6 alkyl-, heteroaryl-C 0 -C 6 alkyl-, cycloalkyl-C 0 -C 6 alkyl-, Heterocyclyl-C 0 -C 6 alkyl-, aryl-C 0 -C 6 alkyl-N(R 3 2 )-, aryl-C(O)-, heteroaryl-C 0 -C 6 alkyl-N ( R 3 2 )-,heterocyclyl-C 0 -C 6 alkyl-N(R 3 2 )-, aryl-O-, heteroaryl-O-, aryl-S-, heteroaryl-S -, aryl-SO 2 -, heteroaryl-SO 2 -, aryl-C(O)N(R 3 2 )-, heteroaryl-C(O)N(R 3 2 )-, heteroaryl -C(H)(SO 2 -heteroaryl)-N(R 3 2 )-; wherein R 3 2 and R 3 3 are independently H or C 1 -C 6 alkyl, or R 3 2 and R 3 3 forms a heterocyclic or heteroaryl group together with the N to which it is attached; and wherein any of the rings described in paragraphs [0045] to [0048] is further substituted, as appropriate, by 1, 2 or 3 independently selected from the group consisting of : alkyl, alkoxy, thioalkoxy, alkyl-SO 2 -, amine, halo, cyano, haloalkyl, hydroxyalkyl, alkoxyalkyl, COOH, alkyl fluorenyl, Alkanoate, NO 2 , hydroxy, haloalkoxy, (R 3 2 )(R 3 3 )N-C 0 -C 6 alkyl-, (R 3 2 )(R 3 3 )N-C 0 - C 6 alkyl-O-, (R 3 2 )(R 3 3 )N-C(O)-, heteroaryl, alkyl-C(O)N(R 3 2 )-, aryl-O- (R 3 2 )(R 3 3 )N-SO 2 -, aryl and (R 3 2 )(R 3 3 )N-alkyl-C(O)N(R 3 2 )-.
"鹵代烴基"為其中一個至所有氫經一或多個鹵基置換之烴基部分。"Halohydrocarbyl" is a hydrocarbyl moiety in which one to all hydrogens are replaced by one or more halo groups.
本文所採用之術語"鹵素"或"鹵基"係指氯、溴、氟或碘。如本文所採用術語"醯基"係指烷羰基或芳羰基取代基。術語"醯胺基"係指於氮原子處被連接之醯胺基(即R－CO－NH－)。術語"胺甲醯基"係指於羰基碳原子處被連接之醯胺基(例如NH2 －CO－)。醯胺基或胺甲醯基取代基之氮原子另外視情況經取代。術語"磺醯胺基"係指由硫或氮原子被連接之磺醯胺取代基。術語"胺基"欲包括NH2 、烷胺基、芳胺基及環胺基。本文所採用術語"脲基"係指經取代或未經取代之尿素部分。The term "halogen" or "halo" as used herein means chloro, bromo, fluoro or iodo. The term "mercapto" as used herein refers to an alkylcarbonyl or arylcarbonyl substituent. The term "ammonium" refers to a guanamine group (ie, R-CO-NH-) that is attached at the nitrogen atom. The term "carbamoyl acyl" refers to a carbonyl carbon atom of the acyl group to be connected (e.g., NH 2 -CO-). The nitrogen atom of the amidino or aminemethanyl substituent is additionally optionally substituted. The term "sulfonamido" refers to a sulfonamide substituent which is attached by a sulfur or nitrogen atom. The term "amino" is intended to include NH 2, alkylamino, arylamino, and cyclic amino. The term "ureido" as used herein refers to a substituted or unsubstituted urea moiety.
本文所用術語"基"意謂包含一或多個未成對電子之化學部分。The term "base" as used herein, means a chemical moiety comprising one or more unpaired electrons.
一經取代之部分係其中一或多個氫已獨立經另一化學取代基置換者。一非限制性實例為，經取代之苯基包括2－氟苯基、3,4－二氯苯基、3－氯－4－氟－苯基、2－氟－3－丙基苯基。另一非限制性實例為，經取代之N－辛基包括2,4－二甲基－5－乙基－辛基及3－環戊基－辛基。包括於該定義者係經氧取代形成羰基(－CO－)之亞甲基(－CH2 －)。A substituted moiety is one in which one or more hydrogens have been independently replaced by another chemical substituent. A non-limiting example is that the substituted phenyl group includes 2-fluorophenyl, 3,4-dichlorophenyl, 3-chloro-4-fluoro-phenyl, 2-fluoro-3-propylphenyl. Another non-limiting example is that the substituted N-octyl group includes 2,4-dimethyl-5-ethyl-octyl and 3-cyclopentyl-octyl. The methylene group (-CH 2 -) which is substituted by the oxygen to form a carbonyl group (-CO-) is included in the definition.
如上文定義之"未經取代"部分(例如未經取代之環烷基、未經取代之雜芳基等)意謂如上文定義之部分，其不具有任何該部分之定義(上文)另外提供之可選性取代基。因此舉例而言，雖然"芳基"包括苯基及經鹵基取代之苯基，但"未經取代之芳基"不包括經鹵基取代之苯基。An "unsubstituted" moiety as defined above (eg, unsubstituted cycloalkyl, unsubstituted heteroaryl, etc.) means a moiety as defined above, which does not have any definition of that moiety (above) Optional substituents are provided. Thus, for example, although "aryl" includes phenyl and halo substituted phenyl, "unsubstituted aryl" does not include halo substituted phenyl.
貫穿本說明書，識別一或多個化學取代基之較佳實施例。較佳實施例之組合亦為較佳。舉例而言，[0080]段描述式(1a)之化合物中Cy之較佳實施例且[0079]段描述式(1a)之化合物之W之較佳實施例。因此其中Cy如[0080]段所描述且W如[0079]段所描述的式(1)之化合物亦涵蓋於本發明之範疇內。Throughout the specification, preferred embodiments of one or more chemical substituents are identified. Combinations of the preferred embodiments are also preferred. For example, paragraph [0080] describes a preferred embodiment of Cy in the compound of formula (1a) and paragraph [0079] describes a preferred embodiment of the compound of formula (1a). Compounds of formula (1) wherein Cy is as described in paragraph [0080] and W is as described in paragraph [0079] are also encompassed within the scope of the invention.
本發明之一些化合物可具有對掌性中心及/或幾何異構中心(E－及Z－異構體)，且應瞭解本發明容納所有該等光學異構體、非對應異構體及幾何異構體。本發明亦包含本文揭示之所有互變異構形式之化合物。當本發明之化合物包括對掌性中心時，本發明容納該等化合物之鏡像異構性純異構體、該等化合物之鏡像異構性富集混合物及該等化合物之外消旋混合物。Some of the compounds of the invention may have a palm center and/or a geometric isomer center (E- and Z-isomers), and it is understood that the invention encompasses all such optical isomers, diastereomers, and geometries. isomer. The invention also encompasses all tautomeric forms of the compounds disclosed herein. When a compound of the invention includes a palmitic center, the invention encompasses the mirror image isomerically pure isomers of such compounds, the mirror image isomerically enriched mixtures of such compounds, and racemic mixtures of such compounds.
本發明之化合物可以活體內可水解酯或活體內可水解醯胺之形式投藥。含有羰基或羥基之本發明化合物的活體內可水解酯例如為在人類或動物體內水解以產生母酸或母醇之醫藥學上可接受之酯。對於羰基合適之醫藥學上可接受之酯包括：C1 － 6 －烷氧甲基酯(例如甲氧甲基)、C1 － 6 －烷醯基氧甲基酯(例如特戊醯氧甲基)、酞基酯、C3 － 8 －環烷氧基羰氧基C1 － 6 －烷基酯(例如1－環己羰基氧乙基)；1,3－二氧雜環戊烯－2－酮基甲基酯(例如5－甲基－1,3－二氧雜環戊烯－2－酮基甲基)；及C1 － 6 －烷氧羰基氧乙基酯(例如1－甲氧羰基氧乙基)且可形成於本發明化合物中之任何羰基處。The compounds of the invention may be administered in the form of hydrolysable esters in vivo or hydrolysable guanamines in vivo. The in vivo hydrolysable ester of the compound of the present invention containing a carbonyl group or a hydroxyl group is, for example, a pharmaceutically acceptable ester which is hydrolyzed in a human or animal body to produce a parent acid or a parent alcohol. Suitable pharmaceutically acceptable esters for carbonyl include: C 1 - 6 - alkoxymethyl esters (eg methoxymethyl), C 1 - 6 -alkyl decyl oxymethyl esters (eg, pentylene oxymethyl) , decyl ester, C 3 - 8 -cycloalkoxycarbonyloxy C 1 - 6 -alkyl ester (eg 1-cyclohexylcarbonyloxyethyl); 1,3-dioxolene - a 2-ketomethyl ester (for example, 5-methyl-1,3-dioxol-2-onemethyl); and a C 1 - 6 - alkoxycarbonyl oxyethyl ester (for example, 1- Methoxycarbonyloxyethyl) can be formed at any carbonyl group in the compounds of the invention.
含有羥基之本發明化合物之活體內可水解酯包括無機酯，諸如磷酸酯及a－醯氧基烷基醚，及作為酯活體內水解產物***產生母羥基的相關化合物。α－醯氧基烷基醚之實例包括乙醯氧基甲氧基及2,2－二甲基丙醯氧基－甲氧基。形成羥基之活體內可水解酯之選擇包括：烷醯基、苯甲醯基、苯乙醯基及經取代之苯甲醯基及苯乙醯基、烷氧羰基(以生成碳酸烷基酯)、二烷基胺甲醯基及N－(N,N－二烷基胺乙基)－N－烷基胺甲醯基(以生成胺基甲酸酯)、N,N－二烷基胺基乙醯基及羧基乙醯基。苯甲醯基上取代基之實例包括自一環氮原子經由一亞甲基連接至苯甲醯基環3－或4－位之嗎啉基及六氫吡嗪基。含有羧基之本發明化合物的活體內可水解醯胺之合適取值(value)例如為：N－C1 － 6 －烷基或N,N－二－C1 － 6 －烷基醯胺，諸如N－甲基、N－乙基、N－丙基、N,N－二甲基、N－乙基－N－甲基或N,N－二乙基醯胺。In vivo hydrolysable esters of the compounds of the invention containing a hydroxy group include inorganic esters such as phosphates and a-methoxyalkyl ethers, and related compounds which are cleaved as in vivo hydrolysates of the ester to form the parent hydroxy group. Examples of the α-methoxyalkyl ether include an ethoxymethoxymethoxy group and a 2,2-dimethylpropoxycarbonyl-methoxy group. The selection of the in vivo hydrolyzable ester forming a hydroxyl group includes: an alkyl fluorenyl group, a benzamidine group, a phenethyl fluorenyl group, a substituted benzyl fluorenyl group, a phenethyl fluorenyl group, and an alkoxycarbonyl group (to form an alkyl carbonate). , dialkylamine methyl sulfhydryl and N-(N,N-dialkylaminoethyl)-N-alkylaminecarbamyl (to form urethane), N,N-dialkylamine Ethyl thiol and carboxyethyl fluorenyl. Examples of the substituent on the benzinyl group include a morpholino group and a hexahydropyrazinyl group which are bonded to the 3- or 4-position of the benzamidine ring via a methylene group from a ring nitrogen atom. Suitable values for the in vivo hydrolysable guanamine of the compound of the invention containing a carboxy group are, for example, N-C 1 - 6 -alkyl or N,N-di-C 1 - 6 -alkyl decylamine, such as N-methyl, N-ethyl, N-propyl, N,N-dimethyl, N-ethyl-N-methyl or N,N-diethylguanamine.
本發明之第一態樣提供組蛋白去乙醯基酶之新穎抑制劑。在第一實施例中，該組蛋白去乙醯基酶之新穎抑制劑由式(1)表示：
及其醫藥學上可接受之鹽，其中X係選自由化學鍵、L、W－L、L－W、L－W－L及L－W'－L－W'所組成之基團，其中Cy為芳基、雜芳基、環烷基或雜環基，其各視情況經取代且各視情況稠合至一或多個芳基或雜芳基環，或稠合至一或多個飽和或部分不飽和之環烷基或雜環基環，該等環各視情況經取代；W在各情況下為S、O、C＝O、－NH－C(＝O)－NH－、－NHSO2 －或N(R9 )，其中R9 係選自由氫、烷基、羥烷基及第三丁氧基羰基所組成之群；W'在各情況下係獨立為化學鍵、S、O或NH；且L在各情況下係獨立為化學鍵或C1 －C4 伸烷基；或Ar為伸芳基或伸雜芳基，其各視情況經取代；q為0或1；且T為NH2 或OH；限制條件為當Cy為萘基，X為－CH2 －，Ar為苯基且q為0或1時T不為OH。And a pharmaceutically acceptable salt thereof, wherein the X group is selected from the group consisting of a chemical bond, L, W-L, L-W, L-W-L, and L-W'-L-W', wherein Cy An aryl, heteroaryl, cycloalkyl or heterocyclyl group, each optionally substituted and optionally fused to one or more aryl or heteroaryl rings, or fused to one or more Or a partially unsaturated cycloalkyl or heterocyclyl ring, each of which is optionally substituted; W in each case S, O, C=O, -NH-C(=O)-NH-, - NHSO 2 - or N(R 9 ), wherein R 9 is selected from the group consisting of hydrogen, alkyl, hydroxyalkyl and tert-butoxycarbonyl; W' is independently a chemical bond, S, O in each case Or NH; and L is independently a chemical bond or a C 1 -C 4 alkyl group in each case; or Ar is an aryl or heteroaryl group, each of which is optionally substituted; q is 0 or 1; Is NH 2 or OH; the restriction is that when Cy is naphthyl, X is -CH 2 -, Ar is phenyl and q is 0 or 1, T is not OH.
如[0059]段之化合物之一些較佳實施例中，q為1且T為NH2 。In some preferred embodiments of the compound of paragraph [0059], q is 1 and T is NH 2 .
[0070]段之實施例之較佳化合物包括彼等Ar為伸苯基且Cy－X－為如下基團者。Preferred compounds of the examples of the paragraph include those wherein Ar is a phenyl group and Cy-X- is a group.
如[0059]段之化合物之一些較佳實施例中，q為0。In some preferred embodiments of the compound of paragraph [0059], q is zero.
一較佳實施例中，本發明之HDAC抑制劑包含具有式(1a)之[0072]段之化合物：
及其醫藥學上可接受之鹽，其中Cy為芳基、雜芳基、環烷基或雜環基，其各視情況經取代，且其各視情況稠合至一或多個芳基或雜芳基環，或稠合至一或多個飽和或部分不飽和環烷基或雜環基環，其各環視情況經取代；W為S、O或N(R9 )，其中R9 為氫或C1 －C6 －烷基；R8 為H或C1 －C4 烷基；且T為NH2 或OH。And a pharmaceutically acceptable salt thereof, wherein Cy is aryl, heteroaryl, cycloalkyl or heterocyclyl, each optionally substituted, and optionally fused to one or more aryl groups or a heteroaryl ring, or fused to one or more saturated or partially unsaturated cycloalkyl or heterocyclyl rings, each optionally substituted; W is S, O or N(R 9 ), wherein R 9 is Hydrogen or C 1 -C 6 -alkyl; R 8 is H or C 1 -C 4 alkyl; and T is NH 2 or OH.
如[0073]段之化合物之一些較佳實施例中，W為NH或S。In some preferred embodiments of the compound of paragraph [0073], W is NH or S.
本發明之較佳化合物及尤其為段[0079]包括彼等Cy係選自由苯基、吡啶基、嘧啶基、吡嗪基、噠嗪基、噻唑基、苯幷噻唑基、苯幷咪唑基及苯幷***基所組成之群者，其各視情況經取代。Preferred compounds of the invention and especially the segment [0079] comprising these Cy groups are selected from the group consisting of phenyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, thiazolyl, benzothiazolyl, benzoimidazolyl and The group consisting of benzotriazole groups is replaced as appropriate.
如[0080]段之較佳化合物包括彼等具有結構1a－1者：
及其醫藥學上可接受之鹽，其中W為NH或S；P、Q、M、G及U係獨立為CH或N，限制條件為P、Q、M、G及U中不多於兩個為N且在含有P、Q、M、G及U之環中，則環S或O不與另一環S或O相鄰；R8 為H或C1 －C4 烷基；且A及B定義如下。And a pharmaceutically acceptable salt thereof, wherein W is NH or S; P, Q, M, G and U are independently CH or N, and the restriction condition is not more than two of P, Q, M, G and U N is in a ring containing P, Q, M, G and U, then ring S or O is not adjacent to another ring S or O; R 8 is H or C 1 -C 4 alkyl; B is defined as follows.
如[0080]段之較佳化合物包括彼等具有結構1a－2者：
及其醫藥學上可接受之鹽，其中W為S或NH；R8 為H或C1 －C4 烷基；且A及B定義如下。And a pharmaceutically acceptable salt thereof, wherein W is S or NH; R 8 is H or C 1 -C 4 alkyl; and A and B are as defined below.
如[0080]段之較佳化合物包括彼等具有結構1a－3者：
及其醫藥學上可接受之鹽，其中W為S或NH，且A及B定義如下。And a pharmaceutically acceptable salt thereof, wherein W is S or NH, and A and B are as defined below.
如[0085]段之較佳化合物包括彼等W為NH者。Preferred compounds such as those in paragraph [0085] include those in which W is NH.
如[0080]段之較佳化合物包括彼等具有結構1a－4者：
及其醫藥學上可接受之鹽，其中W為S或NH；D為N－R1 0 或S，E為N或C－A；R8 及R1 0 係獨立為H或C1 －C4 烷基；且A及B定義如下。And a pharmaceutically acceptable salt thereof, wherein W is S or NH; D is N-R 1 0 or S, E is N or C-A; and R 8 and R 1 0 are independently H or C 1 -C 4 alkyl; and A and B are as defined below.
如[0088]段之較佳化合物包括彼等W為NH者。Preferred compounds as in paragraph [0088] include those in which W is NH.
如[0081]段之較佳化合物包括彼等具有式1a－5者：
及其醫藥學上可接受之鹽。And pharmaceutically acceptable salts thereof.
如[0091]之較佳化合物包括彼等W為NH者。Preferred compounds such as [0091] include those in which W is NH.
如[0073]段及[0079]段之較佳化合物亦包括彼等其中W為NH且Cy為喹噁啉基、鄰苯二甲醯亞胺基或苯幷間二氧雜環戊烯基者，其中各取代基視情況經A及/或B取代，其中A及B定義如下。Preferred compounds such as paragraphs [0073] and [0079] also include those in which W is NH and Cy is quinoxalinyl, phthalimido or benzoquinone dioxolyl Wherein each substituent is substituted with A and/or B as appropriate, wherein A and B are as defined below.
另一較佳實施例中，本發明之HDAC抑制劑包括具有式(1b)之[0059]段之化合物：
及其醫藥學上可接受之鹽，其中Cy為芳基、雜芳基、環烷基或雜環基，其各視情況經取代且其各視情況稠合至一或多個芳基或雜芳基環或稠合至一或多個飽和或部分不飽和環烷基或雜環基環，其中各環視情況經取代；X為L、W－L或L－W，其中W在各情況下為S、O或NH；且L為－CH2 －；Y為N或CH；Z為O、S、NR1 2 或CH2 ；R1 2 為H或C1 －C4 烷基；且T為NH2 或OH。And a pharmaceutically acceptable salt thereof, wherein Cy is aryl, heteroaryl, cycloalkyl or heterocyclyl, each optionally substituted and each optionally fused to one or more aryl or hetero An aryl ring or fused to one or more saturated or partially unsaturated cycloalkyl or heterocyclyl rings, wherein each ring is optionally substituted; X is L, W-L or L-W, wherein W is in each case Is S, O or NH; and L is -CH 2 -; Y is N or CH; Z is O, S, NR 1 2 or CH 2 ; R 1 2 is H or C 1 -C 4 alkyl; Is NH 2 or OH.
如[0095]段之化合物之一些較佳實施例中，T為NH2 。Some preferred embodiments of the compound of [0095] the segment, T is NH 2.
[0095]及[0105]各段之較佳化合物包括彼等其中X為－S－CH2 －、－NH－CH2 －或－CH2 －NH－者。[0095] and [0105] each segment of their preferred compounds include wherein X is -S-CH 2 -, - NH -CH 2 - or by -CH 2 -NH-.
[0095]、[0105]段及0之較佳化合物包括彼等其中Cy為各視情況經取代之芳基或雜芳基者。Preferred compounds of paragraph [0105] and 0 include those in which Cy is optionally substituted aryl or heteroaryl.
[0095]－0各段之較佳化合物包括彼等其中Cy為各視情況經取代之苯基、吡啶基、嘧啶基或苯幷噻唑基者。[0095] Preferred compounds of the various paragraphs include those wherein Cy is optionally substituted phenyl, pyridyl, pyrimidinyl or benzothiazolyl.
[0095]－[0106]各段之較佳化合物包括彼等其中Cy經A及/或B取代者，其中A及B如[0185]段所定義。[00106] Preferred compounds of the various paragraphs include those in which Cy is substituted by A and/or B, wherein A and B are as defined in paragraph [0185].
如[0095]－[0106]各段之較佳化合物包括彼等其中Cy視情況經獨立選自視情況經烷氧基取代之烷氧基、醯基、嗎啉基或苯基之一、二或三個基團取代者。Preferred compounds of the paragraphs [0095] to [0106] include those in which Cy is optionally independently selected from alkoxy, decyl, morpholinyl or phenyl substituted by alkoxy as appropriate. Or three group replacers.
另一較佳實施例中，本發明之HDAC抑制劑包含具有式(1c)之[0059]段之化合物：
及其醫藥學上可接受之鹽，其中Cy為芳基、雜芳基、環烷基或雜環基，其各視情況經取代且其各視情況稠合至一或多個芳基或雜芳基環，或稠合至一或多個飽和或部分不飽和環烷基或雜環基環，其中各環視情況經取代；X為L、W－L或L－W，其中W在各情況下為S、O或NH；且L為－CH2 －；Y為N或CH；Z為O、S、NR1 2 或CH2 ；R1 2 為H或C1 －C4 烷基；且T為NH2 或OH。And a pharmaceutically acceptable salt thereof, wherein Cy is aryl, heteroaryl, cycloalkyl or heterocyclyl, each optionally substituted and each optionally fused to one or more aryl or hetero An aryl ring, or fused to one or more saturated or partially unsaturated cycloalkyl or heterocyclyl rings, wherein each ring is optionally substituted; X is L, W-L or L-W, wherein W is in each case Lower is S, O or NH; and L is -CH 2 -; Y is N or CH; Z is O, S, NR 1 2 or CH 2 ; R 1 2 is H or C 1 -C 4 alkyl; T is NH 2 or OH.
如[0109]段之化合物之一些較佳實施例中，T為NH2 。In some preferred embodiments of the compound of paragraph [0109], T is NH 2 .
如[0109]－[0119]各段之較佳化合物包括彼等其中X為－S－CH2 －、－NH－CH2 －或－CH2 －NH－者。Preferred compounds of the paragraphs [0109] - [0119] include those wherein X is -S-CH 2 -, -NH-CH 2 - or -CH 2 -NH-.
如[0109]－[0120]各段之較佳化合物包括彼等其中Cy為各視情況經取代之芳基或雜芳基者。Preferred compounds of the paragraphs [0109] - [0120] include those in which Cy is optionally substituted aryl or heteroaryl.
如[0109]－[0121]各段之較佳化合物包括彼等其中Cy為各視情況經取代之苯基、吡啶基、嘧啶基或苯幷噻唑基者。Preferred compounds of the paragraphs [0109] to [0121] include those in which Cy is optionally substituted phenyl, pyridyl, pyrimidinyl or benzothiazolyl.
如[0109]－[0122]各段之較佳化合物包括彼等其中Cy為視情況經A及/或B取代者，其中A及B如[0185]段所定義。Preferred compounds of the paragraphs [0109]-[0122] include those in which Cy is optionally substituted by A and/or B, wherein A and B are as defined in paragraph [0185].
如[0109]－[0122]各段之較佳化合物包括彼等Cy視情況經獨立選自視情況經烷氧基取代之烷氧基、鹵代烷氧基、醯基、嗎啉基或苯基之一、二或三個基團取代者。Preferred compounds of the various groups, such as [0109]-[0122], include the alkoxy, haloalkoxy, decyl, morpholinyl or phenyl groups which are optionally independently selected from alkoxy groups as appropriate. One, two or three groups are substituted.
另一較佳實施例中，本發明之HDAC抑制劑包含具有(1d)之[0059]段化合物：
及其醫藥學上可接受之鹽，其中Cy為芳基或雜芳基，其各視情況經取代且其各視情況稠合至一或多個芳基或雜芳基環，或稠合至一或多個飽和或部分不飽和環烷基或雜環基環，其中各環視情況經取代；且T為NH2 或OH。And a pharmaceutically acceptable salt thereof, wherein Cy is aryl or heteroaryl, each optionally substituted and each optionally fused to one or more aryl or heteroaryl rings, or fused to One or more saturated or partially unsaturated cycloalkyl or heterocyclyl rings wherein each ring is optionally substituted; and T is NH 2 or OH.
如[0125]段之化合物之一些較佳實施例中，T為NH2 。In some preferred embodiments of the compound of paragraph [0125], T is NH 2 .
如[0125]－[0129]各段之較佳化合物包括彼等其中Cy為視情況經取代之雜芳基或視情況經取代之雜環基者，該雜芳基或雜環基各視情況稠合至一或多個芳基或雜芳基環，或稠合至一或多個飽和或部分不飽和環烷基或雜環基環，其中各環視情況經取代。Preferred compounds of the paragraphs of [0125]-[0129] include those wherein hetero is optionally substituted heteroaryl or optionally substituted heterocyclyl, optionally, heteroaryl or heterocyclyl. Condensed to one or more aryl or heteroaryl rings, or fused to one or more saturated or partially unsaturated cycloalkyl or heterocyclyl rings, wherein each ring is optionally substituted.
如[0125]－[0130]各段之較佳化合物包括彼等其中Cy為如下基團者：
另一數佳實施例中，本發明之HDAC抑制劑包含具有式(1e)之[0059]段之化合物
及其醫藥學上可接受之鹽，其中Cy為芳基、雜芳基、環烷基或雜環基，其各視情況經取代且各視情況稠合至一或多個芳基或雜芳基，或稠合至一或多個飽和或部分不飽和環烷基或雜環基環，其中各環視情況經取代；且T為NH2 或OH。And a pharmaceutically acceptable salt thereof, wherein Cy is aryl, heteroaryl, cycloalkyl or heterocyclyl, each optionally substituted and optionally fused to one or more aryl or heteroaryl Or, fused to one or more saturated or partially unsaturated cycloalkyl or heterocyclyl rings, wherein each ring is optionally substituted; and T is NH 2 or OH.
如[0132]段之化合物之一些較佳實施例中，T為NH2 。In some preferred embodiments of the compound of paragraph [0132], T is NH 2 .
如[0132]－[0136]各段之較佳化合物包括彼等其中Cy為雜環基或雜芳基者，該雜環基或雜芳基各視情況經取代且各含有至少一個氮原子作為該環部分。Preferred compounds of the paragraphs [0132] to [0136] include those wherein Cy is a heterocyclic group or a heteroaryl group, each of which is optionally substituted and each contains at least one nitrogen atom as The ring part.
如[0132]－[0137]各段之較佳化合物包括彼等其中Cy經一氮原子連接苯基者。Preferred compounds of the various groups such as [0132]-[0137] include those in which Cy is bonded to the phenyl group via a nitrogen atom.
如[0132]－[0137]各段之較佳化合物包括彼等其中Cy為視情況經取代之雜環基者。Preferred compounds of the paragraphs [0132] to [0137] include those in which Cy is optionally substituted heterocyclic.
如[0132]－[0137]及[0139]各段之較佳化合物包括彼等其中Cy為各視情況經取代之哌啶基或六氫吡嗪基者。Preferred compounds of the paragraphs [0132] - [0137] and [0139] include those in which Cy is optionally substituted with piperidinyl or hexahydropyrazinyl.
如[0132]－[0140]各段之較佳化合物包括彼等其中Cy視情況經獨立選自A及B之一或兩個取代基取代者，其中A及B如[0185]段所定義。Preferred compounds of paragraphs such as [0132]-[0140] include those wherein Cy is optionally substituted with one or two substituents independently selected from A and B, wherein A and B are as defined in paragraph [0185].
如[0132]－[0140]各段之較佳化合物包括彼等其中Cy視情況經獨立選自下列基團之一或兩個取代基取代者：
如[0072]段之較佳化合物包括彼等化合物，其中Ar為各視情況經取代之伸苯基、吲哚基或吲哚啉基，且X為不存在、CH2 、－O－CH2 －、－S－CH2 －、－S－C(CH3 )(H)－或－N(R9 )－CH2 －。Preferred compounds as described in paragraph [0072] include such compounds wherein Ar is optionally substituted phenyl, fluorenyl or porphyrin group, and X is absent, CH 2 , -O-CH 2 -, -S-CH 2 -, -S-C(CH 3 )(H)- or -N(R 9 )-CH 2 -.
如[0143]段之較佳化合物包括彼等化合物，其中Ar為吲哚基或吲哚啉基，X為CH2
或－N(R9
)－CH2
－，且Cy為如下基團：
如[0143]段之較佳化合物包括彼等化合物，其中Ar為伸苯基，X為－S－CH2
－或－S－C(CH3
)(H)－，且Cy為：
如[0145]段之較佳化合物包括彼等化合物，其中Y'為H，Y"為：
[0143]段之較佳化合物包括彼等具有式(1f)者：
如[0148]段之較佳化合物包括彼等化合物，其中Cy為雜環基或雜芳基，其各視情況經取代，且其各含有至少一個作為環部分之氮原子。Preferred compounds of paragraph [0148] include those compounds wherein Cy is a heterocyclic or heteroaryl group, each optionally substituted, and each containing at least one nitrogen atom as a ring moiety.
如[0149]段之較佳化合物，包括其他Cy係經由氮原子連結於苯基者。Preferred compounds of paragraph [0149] include those in which other Cy is attached to the phenyl group via a nitrogen atom.
如[0148]段之較佳化合物包括彼等化合物，其中Cy為以下基團：
如[0148]段之較佳化合物包括彼等具有式(1f－1)者：
及其醫藥學上可接受之鹽，其中T為OH或NH2 且A定義如下。And a pharmaceutically acceptable salt thereof, wherein T is OH or NH 2 and A is as defined below.
如[0152]段之較佳化合物包括彼等T為NH2 者。Preferred compounds such as those in paragraph [0152] include those in which T is NH 2 .
如[0143]段之較佳化合物包括彼等化合物，其中Ar為伸苯基，X為－O－CH2
－，且Cy為：
另一較佳實施例中，本發明之HDAC抑制劑包含具有式(1g)之[0059]段之化合物：
及其醫藥學上可接受之鹽，其中Cy為芳基或雜芳基、環烷基或雜環基，其各視情況經取代且其各視情況稠合至一或多個芳基或雜芳基環，或稠合至一或多個飽和或部分不飽和環烷基或雜環基環，其中各環視情況經取代；X為L、W－L或L－W，其中W在各情況下為S、O或NH；且L為－CH2 －；T為NH2 或OH。And a pharmaceutically acceptable salt thereof, wherein Cy is aryl or heteroaryl, cycloalkyl or heterocyclyl, each optionally substituted and each optionally fused to one or more aryl or hetero An aryl ring, or fused to one or more saturated or partially unsaturated cycloalkyl or heterocyclyl rings, wherein each ring is optionally substituted; X is L, W-L or L-W, wherein W is in each case The following is S, O or NH; and L is -CH 2 -; T is NH 2 or OH.
如[0156]段之較佳化合物包括彼等化合物，其中Cy為視情況經取代之雜芳基。更佳Cy為視情況經取代之嘧啶基。亦較佳Cy為經吡啶基取代之嘧啶基。Preferred compounds of paragraph [0156] include those compounds wherein Cy is optionally substituted heteroaryl. More preferably, Cy is a pyrimidinyl group which is optionally substituted. Also preferably, Cy is a pyridyl group substituted with a pyridyl group.
如[0156]段之較佳化合物亦包括彼等其中X為－NH－CH2 －者。Preferred compounds of paragraph [0156] also include those wherein X is -NH-CH 2 -.
如[0156]段之較佳化合物亦包括其中T為NH2 之化合物。Preferred compounds of [0156] segment also include those wherein the compound of 2 T is NH.
如[0059]段之較佳化合物包括彼等具有式(1h)者：
及其醫藥學上可接受之鹽，其中W為S、O或NH且A及B定義如下。And a pharmaceutically acceptable salt thereof, wherein W is S, O or NH and A and B are as defined below.
如[0166]段之較佳化合物包括彼等其中W為NH者。Preferred compounds as in paragraph [0166] include those in which W is NH.
如[0166]段之較佳化合物包括彼等化合物，其中A為視情況經取代之吡啶基或視情況經取代之苯基。Preferred compounds of paragraph [0166] include those compounds wherein A is optionally substituted pyridyl or optionally substituted phenyl.
如[0166]段之較佳化合物包括彼等其中B為H或鹵基者。較佳鹵基為氯。Preferred compounds as in paragraph [0166] include those in which B is H or a halo group. Preferred halo groups are chlorine.
如[0059]段之較佳化合物包括彼等具有式(1i)者：
及其醫藥學上可接受之鹽，其中W為S、O或NH且R4 0 為H或C1 －C6 烷基。And a pharmaceutically acceptable salt thereof, wherein W is S, O or NH and R 4 0 is H or C 1 -C 6 alkyl.
如[0171]段之較佳化合物包括彼等其中W為NH者。Preferred compounds as in paragraph [0171] include those in which W is NH.
如[0171]段之較佳化合物包括彼等其中R4 0 為H者。Preferred compounds such as those in paragraph [0171] include those in which R 4 0 is H.
如[0171]段之較佳化合物包括彼等其中R4 0 為甲基者。Preferred compounds as in paragraph [0171] include those in which R 4 0 is methyl.
另一較佳實施例中，本發明之新穎組蛋白去乙醯基酶抑制劑為式(2)之化合物
及其醫藥學上可接受之鹽，其中Cy2 為各視情況經取代之芳基或雜芳基，且其中芳基及雜芳基各視情況稠合至一或多個芳基或雜芳基環，或稠合至一或多個各視情況經取代之飽和或部分不飽和環烷基或雜環基環；X係選自由下列基團所組成之群：共價鍵、C0 －C4 －烴基、C0 －C4 －烴基－(CO)－C0 －C4 －烴基、C0 －C4 －烴基－(NR7 )－C0 －C4 －烴基、C0 －C4 －烴基－(S)－C0 －C4 －烴基、C0 －C4 －烴基－(O)－C0 －C4 －烴基、C0 －C4 －烴基－(SO)－C0 －C4 －烴基、C0 －C4 －烴基－(SO2 )－C0 －C4 －烴基、C0 －C4 －烴基－(NH)－(CO)－C0 －C4 －烴基、C0 －C4 －烴基－(CO)－(NH)－C0 －C4 －烴基、－NH－CO－NH－、－NH－CS－NH－、－O－CO－O－、－O－CS－O－、－NH－C(NH)－NH－、－S(O)2 －N(R7 )－、－N(R7 )－S(O)2 －、－NH－C(O)－O－及－O－C(O)－NH－，其中R7 係選自由下列基團所組成之群：氫、C1 －C5 －烷基、芳基、芳烷基、醯基、雜環基、雜芳基、SO2 －烷基、SO2 －芳基、CO－烷基、CO－芳基、CO－NH－烷基、CO－NH－芳基、CO－O－烷基及CO－O－芳基，其各視情況經取代，n為0至4，Y為N或CH，T為NH2 或OH。And a pharmaceutically acceptable salt thereof, wherein Cy 2 is an optionally substituted aryl or heteroaryl group, and wherein the aryl group and the heteroaryl group are each fused to one or more aryl or heteroaryl groups, as the case may be. a base ring, or fused to one or more optionally substituted saturated or partially unsaturated cycloalkyl or heterocyclyl rings; X is selected from the group consisting of: covalent bonds, C 0 - C 4 -hydrocarbyl, C 0 -C 4 -hydrocarbyl-(CO)-C 0 -C 4 -hydrocarbyl, C 0 -C 4 -hydrocarbyl-(NR 7 )-C 0 -C 4 -hydrocarbyl, C 0 -C 4 -hydrocarbyl-(S)-C 0 -C 4 -hydrocarbyl, C 0 -C 4 -hydrocarbyl-(O)-C 0 -C 4 -hydrocarbyl, C 0 -C 4 -hydrocarbyl-(SO)-C 0 -C 4 -hydrocarbyl, C 0 -C 4 -hydrocarbyl-(SO 2 )-C 0 -C 4 -hydrocarbyl, C 0 -C 4 -hydrocarbyl-(NH)-(CO)-C 0 -C 4 -hydrocarbyl , C 0 -C 4 -hydrocarbyl-(CO)-(NH)-C 0 -C 4 -hydrocarbyl, -NH-CO-NH-, -NH-CS-NH-, -O-CO-O-, - O-CS-O-, -NH-C(NH)-NH-, -S(O) 2 -N(R 7 )-, -N(R 7 )-S(O) 2 -, -NH-C (O)-O- and -O-C(O)-NH-, which The R 7 group is selected from the group consisting of hydrogen, C 1 -C 5 -alkyl, aryl, aralkyl, fluorenyl, heterocyclic, heteroaryl, SO 2 -alkyl, SO 2 -aryl, CO-alkyl, CO-aryl, CO-NH-alkyl, CO-NH-aryl, CO-O-alkyl and CO-O-aryl, each optionally substituted, n is 0 to 4, Y is N or CH, and T is NH 2 or OH.
式(2)之化合物含有對掌性中心(由星號(＊ )標出)。本發明既容納式(2)之化合物之外消旋混合物又容納其鏡像異構富集混合物，以及式(2)之化合物之鏡像異構純異構體。鏡像異構富集混合物中一種鏡像異構體之含量較佳大於或等於80%，更佳大於90%、95%或98%。The compound of formula (2) contains a palm center (marked by an asterisk ( * )). The present invention contains both the racemic mixture of the compound of formula (2) and its mirror image isomerically enriched mixture, as well as the mirror image isomerized pure isomer of the compound of formula (2). The content of a mirror image isomer in the mirror image-enriched mixture is preferably greater than or equal to 80%, more preferably greater than 90%, 95% or 98%.
基團A及B係相同或不同且係獨立選自H，鹵素，C1
－C4
烷基，包括胺基烷氧基、鹵代烷氧基及雜芳基烷氧基之視情況經取代之烷氧基，烷氧烷基，鹵代烷基，胺基，硝基，烷硫基，醯基胺基，胺甲醯基或以下基團：
較佳可根據流程A說明之程序製備式(1)之化合物，其中Ar為伸吡啶基且X包含－N(R9 )－。以胺RNH2 處理二溴吡啶XIII或XIV分別產生胺基溴吡啶XV或XVI。在一氧化碳下以二乙醯氧基鈀、二苯基膦基二茂鐵、DMF、二異丙基乙胺及苯二胺處理XV或XVI分別產生苯胺醯胺XVII或XVIII。The preferred compound can be described the flow of procedure A of formula (1), the projecting wherein Ar is pyridyl and X comprises -N (R 9) -. Treatment of the dibromopyridine XIII or XIV with the amine RNH 2 yields the amino bromide XV or XVI, respectively. Treatment of XV or XVI with diethoxyphosphoryl palladium, diphenylphosphinoferrocene, DMF, diisopropylethylamine, and phenylenediamine under carbon monoxide produces aniline amines XVII or XVIII, respectively.
在氮氣下DMF中用丙烯酸第三丁酯、二異丙基乙胺、二苄基丙酮鈀及三鄰甲苯基膦(POT)處理XV或XVI分別產生化合物XIX及XX。藉由與二氯甲烷中之三氟乙酸反應，XIX或XX中之酯部分經水解分別產生XXI或XXII中之相應酸部分。以苯二胺、BOP及三乙胺處理酸XXI或XXII分別產生苯胺醯胺XXIII或XXIV。Treatment of XV or XVI with D-butyl acrylate, diisopropylethylamine, palladium dibenzylacetonate and tri-o-tolylphosphine (POT) in DMF under nitrogen afforded compounds XIX and XX, respectively. The ester moiety of XIX or XX is hydrolyzed by reaction with trifluoroacetic acid in dichloromethane to yield the corresponding acid moiety in XXI or XXII, respectively. Treatment of the acid XXI or XXII with phenylenediamine, BOP and triethylamine yields the aniline amine XXIII or XXIV, respectively.
較佳可根據流程B所述之合成途徑製備其中X包含－O－C(O)－NH－之化合物。因此將原醇XXV加入至DMF中偕同羰基二咪唑(CDI)、三乙胺及1,8－二氮雜雙環[5.4.0]十一－7－烯(DBU)之溴苄胺XXVI以產生化合物XXVII。苯胺醯胺XXVIII之製備過程中剩餘合成步驟如上文流程A所述。Preferably, a compound wherein X comprises -O-C(O)-NH- can be prepared according to the synthetic route described in Scheme B. Therefore, the raw alcohol XXV is added to DM in the DMF with carbonyl diimidazole (CDI), triethylamine and 1,8-diazabicyclo [5.4.0] undec-7-ene (DBU) bromobenzylamine XXVI to produce Compound XXVII. The remaining synthetic steps in the preparation of aniline XXVIII are as described in Scheme A above.
較佳可如流程C所述製備化合物，其中X包含－N(R9 )－。使胺XXIX與溴化對溴苄基在DMF中之碳酸鉀存在下反應產生溴苄基胺XXX。在DMF中用硝基丙烯醯基苯胺、二苄基丙酮鈀、POT及二異丙基乙胺處理XXX產生硝基苯胺XXXI。在甲醇及水中用氯化亞錫處理將硝基苯胺XXXI轉化成相應的苯胺醯胺XXXII。Preferred compounds may be prepared as described in Scheme C, where X comprises -N (R 9) -. Reaction of the amine XXIX with bromide bromide in the presence of potassium carbonate in DMF yields the bromobenzylamine XXX. Treatment of XXX with nitropropenyl aniline, dibenzylacetone palladium, POT and diisopropylethylamine in DMF yields nitroaniline XXXI. The nitroaniline XXXI is converted to the corresponding aniline XXXII by treatment with stannous chloride in methanol and water.
在甲酸中用多聚甲醛處理XXXI提供甲胺XXXIII。接著在甲醇及水中用氯化亞錫處理將XXXIII中之硝基甲胺部分轉化成XXXIV中的相應苯胺醯胺部分。Treatment of XXXI with paraformaldehyde in formic acid affords methylamine XXXIII. The nitromethylamine moiety of XXXIII is then partially converted to the corresponding aniline amide moiety of XXXIV by treatment with stannous chloride in methanol and water.
或者，可根據流程D所述之合成途徑製備化合物，其中X包含－N(R9 )－。用氫氯酸處理甲醇中之羧酸XXXV產生酯XXXVI。藉由在60℃下在DMF中用催化劑處理實現將XXXVI中之初級胺部分轉化成XXXVI中之二級胺部分，該催化劑諸如三乙胺、氯化甲氧基苄基、碘化鈉及碳酸鉀。如上文流程A所描述，在DMF中以氫氧化鈉、THF及甲醇繼之以BOP、三乙胺及苯二胺處理將酯XXXVI轉化成苯胺醯胺XXXVII。Alternatively, the compound can be prepared according to the synthetic route of Scheme D, wherein X comprises -N (R 9) -. Treatment of the carboxylic acid XXXV in methanol with hydrochloric acid yields the ester XXXVI. Conversion of the primary amine moiety of XXXVI to the secondary amine moiety of XXXVI by treatment with a catalyst in DMF at 60 ° C, such as triethylamine, methoxybenzyl chloride, sodium iodide and carbonic acid Potassium. The ester XXXVI is converted to the aniline XXXVII by treatment with BOP, triethylamine and phenylenediamine in DMF with sodium hydroxide, THF and methanol followed by treatment as described above.
較佳可根據流程E說明之程序製備化合物，其中X包含或－C(O)－NH－。將胺68加入至鹵代芳基化合物XXXVIII或XXXIX及DMF中之碳酸鉀中分別提供芳基胺XL或XLI。接著用類似於彼等上文流程A－D所述之程序製備苯胺醯胺XLII或XLIII。Preferably, the compound can be prepared according to the procedure described in Scheme E, wherein X comprises Or -C(O)-NH-. Amine 68 is added to the halogenated aryl compound XXXVIII or XXXIX and potassium carbonate in DMF to provide an arylamine XL or XLI, respectively. The aniline amine XLII or XLIII is then prepared using procedures similar to those described above for Schemes A-D.
較佳如流程F所述製備諸如XLVII及XLIX之化合物。將二溴吡啶與二胺基乙烷組合產生胺XLIV。在甲醇及水中用靛紅酸酐LV處理胺XLIV，隨後在甲酸中回流產生化合物XLVI。用苄胺基二乙酸與乙酸酐之反應產物處理胺XLIV提供化合物XLVIII。接著由類似於彼等上文流程A－E所述之程序將溴代吡啶胺XLVI及XLVIII分別轉化成相應的二烯苯胺醯胺XLVII及XLIX。Compounds such as XLVII and XLIX are preferably prepared as described in Scheme F. Dibromopyridine is combined with diaminoethane to produce the amine XLIV. Treatment of the amine XLIV with isatoic anhydride LV in methanol and water followed by reflux in formic acid afforded compound XLVI. Treatment of the amine XLIV with the reaction product of benzylaminodiacetic acid and acetic anhydride provides compound XLVIII. The bromopyridinium XLVI and XLVIII are then converted to the corresponding dienylamine amines XLVII and XLIX, respectively, by procedures analogous to those described above in Schemes A-E.
較佳可根據流程G所述之合成途徑製備諸如LIV之化合物。用胺基部滿及二異丙基乙胺處理三氯三嗪產生二氯胺基三嗪L。用溴苄基胺及二異丙基乙胺處理產生二胺基氯三嗪LI。添加氨氣及二噁烷提供三胺基三嗪LII。用經保護之丙烯醯基苯胺、三乙胺、POT及二苄基丙酮鈀處理，接著產生二烯苯胺醯胺LIII，其經三氟乙酸去保護提供最終產物LIV。Preferably, a compound such as LIV can be prepared according to the synthetic route described in Scheme G. Treatment of trichlorotriazine with an amine base and diisopropylethylamine produces dichloroaminotriazine L. Treatment with bromobenzylamine and diisopropylethylamine yields the diaminochlorotriazine LI. The addition of ammonia gas and dioxane provides the triamine triazine LII. Treatment with protected acryloylaniline, triethylamine, POT and dibenzylacetone palladium, followed by the production of the dienylbenzamide LIII, which is deprotected with trifluoroacetic acid to provide the final product LIV.
較佳可根據流程H說明之程序製備式(1)之化合物，其中Ar為伸喹啉基(quinolylene)且X包含－N(R9 )－。用三氟甲磺酸酐處理吡啶中之二羥基喹啉LV與二甲基胺基吡啶(DMAP)提供雙(三氟甲磺醯氧基)－喹啉LVI。用對甲氧苄胺處理LVI產生胺基喹啉LVII。接著用類似於彼等上文描述之程序製備苯胺醯胺LVIII及LIX。Scheme H may be preferred compound described procedure for preparation of formula (1) of, wherein Ar is quinolinyl extension (quinolylene) and X comprises -N (R 9) -. Treatment of dihydroxyquinoline LV and dimethylaminopyridine (DMAP) in pyridine with trifluoromethanesulfonic anhydride provides bis(trifluoromethanesulfonyloxy)-quinoline LVI. Treatment of LVI with p-methoxybenzylamine yields the aminoquinoline LVII. The aniline amines LVIII and LIX are then prepared using procedures similar to those described above.
較佳如流程I所述製備化合物，其中X包含一硫原子。用類似於彼等上文流程D所述之程序將溴化物LX轉化成二芳基酯LXI。藉由用諸如氫氧化鋰之氫氧化物鹼處理將酯LXI轉化成相應酸LXIV。或者，可用DMF中之氯乙基嗎啉(morphonline)、碘化鈉、碳酸鉀、三乙胺及碘化四丁基銨(TBAI)處理酯LXI產生酯LXIII，接著將LXIII轉化成酸LXIV。藉由在BOP試劑、三乙胺及二甲基甲醯胺(DMF)存在下藉由用1,2－苯二胺處理該酸實現酸LXIV至苯胺醯胺LXV的轉化。Preferably, a compound is prepared as described in Scheme I wherein X comprises a sulfur atom. The bromide LX is converted to the diaryl ester LXI by a procedure similar to that described in Scheme D above. The ester LXI is converted to the corresponding acid LXIV by treatment with a hydroxide base such as lithium hydroxide. Alternatively, ester LXI can be treated with morphonline, sodium iodide, potassium carbonate, triethylamine, and tetrabutylammonium iodide (TBAI) in DMF to yield ester LXIII, followed by conversion of LXIII to acid LXIV. Conversion of the acid LXIV to the aniline amine LXV is achieved by treating the acid with 1,2-phenylenediamine in the presence of BOP reagent, triethylamine and dimethylformamide (DMF).
或者，可根據流程J說明之程序製備化合物，其中X包含硫原子。用類似於彼等上文所述之程序製備硫基苯胺醯胺LXVIII。Alternatively, the compound can be prepared according to the procedure described in Scheme J, wherein X contains a sulfur atom. Thioanilinamide LXVIII was prepared using procedures similar to those described above.
較佳可根據流程K所述之合成途徑製備化合物，其中X包含－N(R9 )－。根據類似於彼等上文描述之合成步驟製備胺基苯胺醯胺LXXI。Preferred compounds can be prepared according to the synthetic route of Scheme K, wherein X comprises -N (R 9) -. The aminoanilinamine LXXI was prepared according to a synthetic procedure similar to that described above.
可如流程L所述製備化合物，其中X包含硫原子。使苯二胺與二第三丁基二碳酸酯反應，繼之以與碘苯甲酸、二甲基胺丙基乙基碳化二醯亞胺、羥基苯幷***及三乙胺反應以提供經保護之苯胺醯胺LXXII。用類似於彼等上文所述之程序將LXXII之碘化物部分轉化成LXXIII之甲酯部分。用諸如氫化二異丁基鋁(DIBAL－H)之還原劑將LXXIII之甲酯部分轉化成LXXIV之羥基部分。添加雜環基氫硫基化合物Het－SH與三苯膦及偶氮二羧酸二乙酯將LXXIV之羥基部分轉化成LXXV之硫基部分。經三氟乙酸去保護LXXV以產生硫基苯胺醯胺LXXVI。Compounds can be prepared as described in Scheme L wherein X contains a sulfur atom. The phenylenediamine is reacted with di-tert-butyl dicarbonate, followed by reaction with iodobenzoic acid, dimethylaminopropyl carbodiimide, hydroxybenzotriazole and triethylamine to provide Protected aniline amine LXXII. The iodide moiety of LXXII is converted to the methyl ester portion of LXXIII using procedures similar to those described above. The methyl ester moiety of LXXIII is converted to the hydroxyl moiety of LXXIV with a reducing agent such as diisobutylaluminum hydride (DIBAL-H). The heterocyclic thiol compound Het-SH is added with triphenylphosphine and diethyl azodicarboxylate to convert the hydroxyl moiety of LXXIV to the thio moiety of LXXV. The LXXV was deprotected with trifluoroacetic acid to yield the thioanilinamide LXXVI.
可根據流程M所述之合成途徑製備化合物，其中X為化學鍵。因此，用芳基酸、苯、乙醇、碳酸鈉水溶液及三苯基膦鈀處理氯代芳基苯胺醯胺LXXVII以產生二芳基苯胺醯胺LXXVIII。Compounds can be prepared according to the synthetic route described in Scheme M, wherein X is a chemical bond. Therefore, using aryl The chloroarylanilide amide LXXVII is treated with an acid, benzene, ethanol, aqueous sodium carbonate solution and triphenylphosphine palladium to yield the diarylanilinamide LXXVIII.
較佳可根據流程N說明之程序製備諸如LXXXI之化合物。因此，用對胺甲基苯甲酸處理乙酸中之苯－1,2－甲醛LXXIX產生苯甲酸LXXX。藉由用羥基苯幷***、二氯乙烯及苯二胺進行處理將酸LXXX轉化成相應的苯胺醯胺LXXXI。Preferably, a compound such as LXXXI can be prepared according to the procedure described in Scheme N. Thus, treatment of phenyl-1,2-formaldehyde LXXIX in acetic acid with p-aminomethylbenzoic acid produces benzoic acid LXXX. The acid LXXX is converted to the corresponding aniline amine LXXXI by treatment with hydroxybenzotriazole, dichloroethylene and phenylenediamine.
較佳可根據流程O說明之程序製備諸如LXXXVI及LXXXIX之化合物。使鄰苯二甲酸酐LXXXV及對胺甲基苯甲酸在乙酸中組合產生中間體羧酸，用類似於彼等上文所述之程序將該中間體羧酸轉化成苯胺醯胺LXXXVI。Preferably, compounds such as LXXXVI and LXXXIX can be prepared according to the procedures described in Scheme O. The phthalic anhydride LXXXV and p-aminomethylbenzoic acid are combined in acetic acid to yield the intermediate carboxylic acid which is converted to the aniline amine LXXXVI using procedures similar to those described above.
將4－(2－胺乙基)苯酚添加至乙酸中之鄰苯二甲酸酐LXXXV產生羥基化合物LXXXVII。藉由用氫化鈉、THF、DMF及雙三氟甲磺酸苯胺酯進行處理將LXXXVII之羥基轉化成LXXXVIII之三氟甲磺酸酯基。根據類似於彼等流程3之描述處理LXXXVIII產生苯胺醯胺LXXXIX。Addition of 4-(2-aminoethyl)phenol to phthalic anhydride LXXXV in acetic acid yields hydroxy compound LXXXVII. The hydroxyl group of LXXXVII is converted to the triflate group of LXXXVIII by treatment with sodium hydride, THF, DMF and bis(trifluoromethanesulfonate). Treatment of LXXXVIII according to a description similar to that of Scheme 3 yields the aniline amine LXXXIX.
較佳可根據流程P所述之合成途徑製備諸如XCI－XCVI 之化合物。以水及三乙胺中之對胺甲基苯甲酸繼之以甲酸處理靛紅酸酐XC 產生中間體羧酸，其用類似於彼等上文描述之程序轉化成苯胺醯胺XCI 。Preferably, a compound such as XCI-XCVI can be prepared according to the synthetic route described in Scheme P. Treatment of isatoic anhydride XC with water and triethylamine in the amine methyl benzoic acid followed by formic acid to yield the intermediate carboxylic acid which was converted to the aniline amine XCI using procedures similar to those described above.
或者，以水及三乙胺中之對胺甲基苯甲酸繼之以氫氯酸及亞硝酸鈉處理衣托酸(isatoic acid)XC產生中間體羧酸，其用類似於彼等上文描述之程序轉化成苯胺醯胺XCII。Alternatively, the treatment of the isatoic acid XC with an amine methyl benzoic acid in water and triethylamine followed by hydrochloric acid and sodium nitrite produces an intermediate carboxylic acid similar to that described above. The procedure is converted to the aniline amine XCII.
或者以水及三乙胺中之對胺甲基苯甲酸處理衣托酸XC產生苯甲酸XCIII。用氫氧化鈉、二噁烷、氯甲酸甲酯及甲醇處理XCIII產生中間體喹唑啉二酮羧酸，接著將其酸部分用類似於彼等上文描述之程序轉化成XCIV之苯胺醯胺部分。或者，用碳酸鉀及甲基碘處理DMF中之該中間體喹唑啉二酮羧酸產生中間體苯甲酸甲酯，其藉由用氫氧化鈉、甲醇及水進行處理轉化成中間體苯甲酸。接著用類似於彼等上文描述之程序將該苯甲酸轉化成相應苯胺醯胺XCV。Alternatively, the benzoic acid XCIII is produced by treating the enamel acid XC with water and triethylamine in the amine methylbenzoic acid. Treatment of XCIII with sodium hydroxide, dioxane, methyl chloroformate and methanol yields the intermediate quinazolinedione carboxylic acid, which is then converted to the aniline amine of XCIV using procedures similar to those described above. section. Alternatively, the intermediate quinazolinedione carboxylic acid in DMF is treated with potassium carbonate and methyl iodide to produce an intermediate methyl benzoate which is converted to the intermediate benzoic acid by treatment with sodium hydroxide, methanol and water. . This benzoic acid is then converted to the corresponding aniline oxime XCV using procedures similar to those described above.
或者，用乙酸酐繼之以乙酸處理XCIII產生中間體羧酸，其用類似於彼等上文描述之程序將中間體羧酸轉化成苯胺醯胺XCVI。Alternatively, treatment of XCIII with acetic anhydride followed by acetic acid yields the intermediate carboxylic acid which is converted to the aniline amine XCVI using procedures similar to those described above.
較佳可如流程Q所述製備諸如C之化合物。以二氯甲烷中之硫羰基二咪唑、繼之以氫氧化銨處理烷基胺XCVII產生硫脲XCVIII。在二噁烷及N－溴代丁二醯亞胺中用甲氧基丙烯酸甲酯處理硫脲XCVIII產生噻唑酯IC。用類似於彼等上文所述之程序將酯IC轉化成相應苯胺醯胺C。Preferably, a compound such as C can be prepared as described in Scheme Q. Treatment of the alkylamine XCVII with thiocarbonyldiimidazole in dichloromethane followed by ammonium hydroxide produces thiourea XCVIII. Treatment of thiourea XCVIII with methyl methacrylate in dioxane and N-bromosuccinimide yields the thiazol ester IC. The ester IC is converted to the corresponding aniline amine C using procedures similar to those described above.
可根據R中所述之合成途徑製備化合物，其中X為化學鍵且Cy具有一胺基取代基。因此，根據類似於彼等上文描述之程序處理經保護之碘芳基苯胺醯胺CI以產生二芳基苯胺醯胺CII。藉由以初級胺及三乙醯氧基硼氫化鈉繼之以冰乙酸進行處理使CII中之醛部分轉化成相應二級胺部分。用類似於彼等上文所述之程序將所得化合物去保護產生CIII。Compounds can be prepared according to the synthetic routes described in R wherein X is a chemical bond and Cy has an amine substituent. Thus, the protected iodoarylanilide amine CI is treated according to procedures similar to those described above to produce the diarylanilinamide CII. The aldehyde moiety in CII is converted to the corresponding secondary amine moiety by treatment with a primary amine and sodium triethoxy borohydride followed by glacial acetic acid. The resulting compound is deprotected to produce CIII using procedures similar to those described above.
可如流程S所述製備化合物，其中X包含伸烷基部分。用氯化三苯基膦鈀、碘化亞銅及1－乙炔基環己胺處理經保護之碘芳基苯胺醯胺CI產生炔芳基苯胺醯胺CIV。將CIV中初級胺部分轉化成相應二級胺且用類似於彼等上文描述之程序將胺部分去保護以產生CV。Compounds can be prepared as described in Scheme S wherein X comprises an alkylene moiety. Treatment of the protected iodoarylanilide amine CI with triphenylphosphine palladium chloride, cuprous iodide and 1-ethynylcyclohexylamine produces an alkyne aryl anilide CIV. The primary amine moiety in CIV is converted to the corresponding secondary amine and the amine moiety is deprotected to produce CV using procedures similar to those described above.
較佳可根據流程T所述之合成途徑製備諸如CVIII之化合物。在二異丙基乙胺存在下用甲基－4－胺基苯甲酸酯處理二氯胺基三嗪CVI產生二胺基三嗪CVII。加入氨氣及二噁烷，隨後進行皂化及縮胺酸偶合(peptide coupling)產生CVIII。Preferably, a compound such as CVIII can be prepared according to the synthetic route described in Scheme T. Treatment of the dichloroaminotriazine CVI with methyl-4-aminobenzoic acid ester in the presence of diisopropylethylamine produces the diaminotriazine CVII. Ammonia gas and dioxane are added, followed by saponification and peptide coupling to produce CVIII.
較佳可根據流程U所述之合成途徑製備諸如CX之化合物。三氯胺基三嗪與各種烷基溴化鎂之格林納(Grignard)反應，隨後在二異丙基乙胺存在下用甲基－4－胺基苯甲酸酯處理產生烷胺基三嗪CIX。接著用類似於彼等上文描述之合成方法將酯CIX轉化成相應苯胺醯胺CX。Preferably, a compound such as CX can be prepared according to the synthetic route described in Scheme U. Trichloroaminotriazine is reacted with various alkylmagnesium bromide in Grignard, followed by treatment with methyl-4-aminobenzoate in the presence of diisopropylethylamine to give the alkylaminotriazine CIX. The ester CIX is then converted to the corresponding anilineamine CX using a synthetic procedure similar to that described above.
如流程V所示，胺化二氯三嗪產生CXI。用乙烯基錫烷進行Stille偶合提供CXII。以經保護之碘醯苯胺、三乙胺、POT及二苄基丙酮鈀處理接著產生苯胺醯胺，其經三氟乙酸去保護提供烯烴CXIII。烯烴氫化產生最終化合物CXIV。As shown in Scheme V, the aminated dichlorotriazine produces CXI. Stille coupling with vinylstannane provides CXII. Treatment with protected iodonium anilide, triethylamine, POT and dibenzylacetone palladium then yields the aniline amine which is deprotected with trifluoroacetic acid to provide the olefin CXIII. Hydrogenation of the olefin produces the final compound CXIV.
較佳可根據流程W所述之合成途徑製備諸如CXVIII之化合物。用三溴化硼處理甲氧胺基苯幷噻唑產生相應酸CXV。在偶氮二羧酸二乙酯及三苯膦存在下使用羥乙基嗎啉進行遠藤(Mitsunobu)反應產生胺CXVI。用苯基矽烷及錫催化劑還原胺化甲基－4－甲醯基苯甲酸酯產生酯CXVII。類似於彼等上文描述進行皂化隨後常見縮胺酸偶合提供所要之醯苯胺CXVIII。Preferably, a compound such as CXVIII can be prepared according to the synthetic route described in Scheme W. Treatment of methoxyaminobenzothiazole with boron tribromide produces the corresponding acid CXV. The Mitsunobu reaction was carried out using hydroxyethylmorpholine in the presence of diethyl azodicarboxylate and triphenylphosphine to give the amine CXVI. Reductive amination of methyl-4-carbamidobenzoate with phenyl decane and a tin catalyst yields ester CXVII. Saponification is carried out analogously to their above description followed by the usual amino acid coupling to provide the desired indole amide CXVIII.
用硫化氫處理4－甲基氰基苯甲酸產生CXIX，其在1,3－二氯丙酮存在下經環化作用產生CXX。用標準條件用嗎啉處理隨後縮胺酸偶合產生CXXI。Treatment of 4-methylcyanobenzoic acid with hydrogen sulfide produces CXIX which is cyclized to give CXX in the presence of 1,3-dichloroacetone. Treatment with morpholine under standard conditions followed by amino acid coupling yielded CXXI.
較佳可根據合成流程Y製備諸如CXXIII及CXXVII之化合物。用苯肼存在下之溴甲基苯甲酸甲酯及NaOMe連續處理乙醯基丙酮隨後進行皂化產生中間體酸CXXII。該材料與1,2－二胺基苯以標準形式偶合產生CXXIII。Compounds such as CXXIII and CXXVII are preferably prepared according to the synthetic scheme Y. The intermediate acid CXXII is produced by continuous treatment of methyl bromomethylbenzoate with NaOMe in the presence of phenylhydrazine and subsequent saponification. This material is coupled with 1,2-diaminobenzene in standard form to give CXXIII.
以NaOMe存在下之溴甲基苯甲酸甲酯及AcOH－HCl之1：1(濃度)混合物連續處理乙醯基丙酮產生中間體酸CXXIV。在鹼存在下該酮－酸與硫及丙二腈反應產生噻吩CXXV，用標準程序將其轉化成所要CXXVII。The acetonitrile was continuously treated with a 1:1 (concentration) mixture of methyl bromomethylbenzoate and AcOH-HCl in the presence of NaOMe to give the intermediate acid CXXIV. The keto-acid is reacted with sulfur and malononitrile in the presence of a base to yield the thiophene CXXV which is converted to the desired CXXVII using standard procedures.
較佳可根據合成流程Z製備諸如CXXX之化合物。用羥基胺處理4－氰甲基苯甲酸產生醯胺肟CXXVIII，其經乙酸酐處理轉化成噁二唑CXXIX。後者以標準形式與1,2－二胺基苯偶合產生CXXX。Preferably, a compound such as CXXX can be prepared according to the synthetic scheme Z. Treatment of 4-cyanomethylbenzoic acid with hydroxylamine yields amidoxime CXXVIII which is converted to the oxadiazole CXXIX by treatment with acetic anhydride. The latter is coupled to 1,2-diaminobenzene in a standard form to give CXXX.
較佳可根據流程AA所述之合成途徑製備諸如CXXXIII之化合物。以亞硫醯氯處理4－甲醯基苯甲酸產生醯基氯，其與經保護之苯胺偶合產生CXXXI。用苯基矽烷及錫催化劑還原胺化二甲氧基苯胺產生經保護之苯胺CXXXII。用異氰酸酯處理隨後經三氟乙酸去保護提供脲基醯苯胺CXXXIII。Preferably, a compound such as CXXXIII can be prepared according to the synthetic route described in Scheme AA. Treatment of 4-mercaptobenzoic acid with sulfinium chloride produces mercapto chloride which is coupled to the protected aniline to give CXXXI. Reductive amination of dimethoxyaniline with phenyl decane and a tin catalyst produces protected aniline CXXXII. Treatment with isocyanate followed by deprotection with trifluoroacetic acid affords the ureidoaniline CXXXIII.
第二態樣中，本發明提供醫藥組合物，其包含本發明之組蛋白去乙醯基酶抑制劑及醫藥學上可接受之載劑、賦形劑或稀釋劑。本發明之化合物可藉由此項技術中熟知之任何方法加以調配且可製備為針對任何投藥途徑，包括(不限於)非經腸、口服、舌下、經皮、局部、鼻內、氣管內或直腸內。在特定較佳實施例中，本發明之化合物在醫院環境中經靜脈內投藥。在特定其它較佳實施例中較佳經口服途徑投藥。In a second aspect, the invention provides a pharmaceutical composition comprising a histone deacetylase inhibitor of the invention and a pharmaceutically acceptable carrier, excipient or diluent. The compounds of the invention may be formulated by any of the methods well known in the art and may be prepared for any route of administration including, without limitation, parenteral, oral, sublingual, transdermal, topical, intranasal, intratracheal. Or in the rectum. In a particularly preferred embodiment, the compounds of the invention are administered intravenously in a hospital setting. In certain other preferred embodiments, it is preferred to administer the drug by the oral route.
載劑特徵應視投藥途徑而定。本文所用術語"醫藥學上可接受的"意謂與諸如細胞、細胞培養物、組織或有機體之生物系統相容的無毒材料，且該材料不妨礙活性成份生物活性之效力。因此、本發明之組合物除抑制劑之外還可含有稀釋劑、填充劑、鹽、緩衝劑、穩定劑、增溶劑及其它此項技術中熟知之材料。製備醫藥學上可接受之調配物例如描述於Remington's Pharmaceutical Sciences，第18版，A.Gennaro編，Mack Publishing Co.,Easton,PA,1990。The characteristics of the carrier should depend on the route of administration. The term "pharmaceutically acceptable" as used herein, means a non-toxic material that is compatible with a biological system such as a cell, cell culture, tissue or organism, and which does not interfere with the effectiveness of the biological activity of the active ingredient. Thus, the compositions of the present invention may contain, in addition to the inhibitor, diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art. Preparation of pharmaceutically acceptable formulations is described, for example, in Remington's Pharmaceutical Sciences, 18th Ed., edited by A. Gennaro, Mack Publishing Co., Easton, PA, 1990.
本文所用術語醫藥學上可接受之鹽係指保留上文確定之化合物之所要生物活性且呈現最小或無所要毒理作用的鹽。該等鹽之實例包括(但不限於)：與無機酸(例如氫氯酸、氫溴酸、硫酸、磷酸、硝酸及其類似物)形成之酸加成鹽，與諸如乙酸、草酸、酒石酸、丁二酸、蘋果酸、抗壞血酸、苯甲酸、鞣酸、雙羥萘酸、海藻酸、聚麩胺酸、萘磺酸、萘二磺酸及聚半乳糖醛酸之有機酸形成之鹽。該等化合物亦可作為熟悉此項技術者已知之醫藥學上可接受之四級鹽投藥，該等四級鹽特別包括式－NR＋Z－之四級銨鹽，其中R為氫、烷基或苄基，且Z為平衡離子，包括氯離子、溴離子、碘離子、－O－烷基、甲苯磺酸根、甲基磺酸根、磺酸根、磷酸根或羧酸根(諸如苯甲酸根、丁二酸根、乙酸根、羥乙酸根、順丁烯二酸根、蘋果酸根、檸檬酸根、酒石酸根、抗壞血酸根、苯甲酸根、肉桂酸根、扁桃酸根、苄酸根(benzyloate)及二苯基乙酸根)。The term pharmaceutically acceptable salt as used herein refers to a salt which retains the desired biological activity of the compound identified above and which exhibits minimal or no toxicological effects. Examples of such salts include, but are not limited to, acid addition salts with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like, and such as acetic acid, oxalic acid, tartaric acid, A salt formed from an organic acid of succinic acid, malic acid, ascorbic acid, benzoic acid, citric acid, pamoic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, naphthalenedisulfonic acid, and polygalacturonic acid. The compounds may also be administered as pharmaceutically acceptable quaternary salts known to those skilled in the art, and the quaternary salts include, in particular, a quaternary ammonium salt of the formula -NR+Z- wherein R is hydrogen, alkyl or benzyl. And Z is a counter ion, including chloride, bromide, iodide, -O-alkyl, tosylate, methanesulfonate, sulfonate, phosphate or carboxylate (such as benzoate, succinate) , acetate, glycolate, maleate, malate, citrate, tartrate, ascorbate, benzoate, cinnamate, mandelate, benzyloate and diphenylacetate).
醫藥學上可接受之載劑或稀釋劑中包括之活性化合物具有足夠向患者輸送治療有效劑量而不對所治療之患者引起嚴重毒性作用之量。用於上文提及之所有條件的活性化合物之較佳劑量係在每天約0.01至300 mg/kg、較佳每天0.1至100 mg/Kg之範圍內，更一般每天每公斤受者體重0.5至約25 mg。一典型局部劑量應為在合適載劑中0.01至3% wt/wt範圍內。醫藥學上可接受之衍生物的有效劑量範圍可基於欲輸送之母化合物重量計算。若該衍生物自身呈現活性，可使用衍生物之重量如上估算，或藉由熟習此項技術者已知之其它方法估算有效劑量。The active compound included in a pharmaceutically acceptable carrier or diluent has an amount sufficient to deliver a therapeutically effective amount to a patient without causing a serious toxic effect on the patient being treated. Preferred dosages of the active compounds for all of the conditions mentioned above are in the range of from about 0.01 to 300 mg/kg per day, preferably from 0.1 to 100 mg/kg per day, more typically from 0.5 to 0.5 per kg of body weight per day. About 25 mg. A typical topical dose should be in the range of 0.01 to 3% wt/wt in a suitable carrier. The effective dosage range for a pharmaceutically acceptable derivative can be calculated based on the weight of the parent compound to be delivered. If the derivative itself is active, the weight of the derivative can be estimated as above, or the effective dose can be estimated by other methods known to those skilled in the art.
在第三態樣中，本發明提供一種抑制細胞中組蛋白去乙醯基酶之方法，該方法包含使需要抑制組蛋白去乙醯基酶之細胞與本發明之組蛋白去乙醯基酶抑制劑接觸。由於本發明之化合物抑制組蛋白去乙醯基酶，因此其係活體外研究組蛋白去乙醯基酶在生物過程中之影響的可用研究工具。此外，本發明之化合物選擇性抑制HDAC之特定異構體。In a third aspect, the present invention provides a method for inhibiting histone deacetylase in a cell, the method comprising: a cell which requires inhibition of histone deacetylase and a histone deacetylase of the present invention Inhibitor contact. Since the compounds of the present invention inhibit histone deacetylase, they are useful research tools for studying the effects of histone deacetylase in biological processes in vitro. Furthermore, the compounds of the invention selectively inhibit specific isomers of HDAC.
可使用已知方法實現組蛋白去乙醯基酶酶活性之量測。舉例而言，Yoshida等人，J.Biol.Chem.,265：17174－17179(1990)描述藉由偵測經曲古柳菌素A處理之細胞中經乙醯化之組蛋白來評估組蛋白去乙醯基酶酶活性。Taunton等人，Science,272：408－411(1996)類似地描述使用內因性及重組性HDAC－1量測組蛋白去乙醯基酶酶活性之方法。The measurement of histone deacetylase enzymatic activity can be achieved using known methods. For example, Yoshida et al, J. Biol. Chem., 265: 17174-17179 (1990) describe the assessment of histones by detecting acetylated histones in cells treated with trichostatin A. Deacetylase enzymatic activity. Taunton et al, Science, 272: 408-411 (1996) similarly describe a method for measuring histone deacetylase enzymatic activity using endogenous and recombinant HDAC-1.
一些較佳實施例中，組蛋白去乙醯基酶抑制劑與細胞中所有組蛋白去乙醯基酶相互作用並降低其活性。本發明此態樣之一些其它較佳實施例中，組蛋白去乙醯基酶抑制劑與細胞中少於全部之組蛋白去乙醯基酶相互作用並降低其活性。特定較佳實施例中，該抑制劑與一種組蛋白去乙醯基酶(例如HDAC－1)或組蛋白去乙醯基酶之亞群(例如HDAC－1、HDAC－2及HDAC－3)相互作用並將其活性降低至比其它組蛋白去乙醯基酶更大之程度。當該抑制劑優先降低組蛋白去乙醯基酶亞群之活性時，該亞群中各成員活性降低量可相同或不同。如下論述，特定尤其較佳之組蛋白去乙醯基酶抑制劑為彼等與涉及腫瘤發生之組蛋白去乙醯基酶相互作用並減少其酶活性者。特定其它較佳組蛋白去乙醯基酶抑制劑與真菌組蛋白去乙醯基酶相互作用並減少其酶活性。In some preferred embodiments, the histone deacetylase inhibitor interacts with and reduces the activity of all histone deacetylases in the cell. In some other preferred embodiments of this aspect of the invention, the histone deacetylase inhibitor interacts with less than all of the histone deacetylase in the cell and reduces its activity. In a particularly preferred embodiment, the inhibitor is associated with a histone deacetylase (eg, HDAC-1) or a subgroup of histone deacetylase (eg, HDAC-1, HDAC-2, and HDAC-3) Interact and reduce its activity to a greater extent than other histone deacetylases. When the inhibitor preferentially reduces the activity of the histone deacetylase subgroup, the amount of activity reduction of each member in the subgroup may be the same or different. As discussed below, certain particularly preferred histone deacetylase inhibitors are those which interact with histone deacetylase involved in tumorigenesis and reduce its enzymatic activity. Certain other preferred histone deacetylase inhibitors interact with the fungal histone deacetylase and reduce their enzymatic activity.
本發明第三態樣之方法較佳引起對所接觸細胞之細胞增殖的抑制作用。短語"抑制細胞增殖"用於表示組蛋白去乙醯基酶抑制劑延遲與抑制劑接觸之細胞相較於未接觸之細胞而言生長的能力。藉由使用Coulter細胞計數器(Coulter,Miami,FL)或血細胞計數器計數經接觸及未經接觸細胞來進行細胞增殖的評估。當細胞處於固體生長(例如固體腫瘤或器官)中時，可藉由用測徑器量測生長且將經接觸細胞與未經接觸細胞之生長尺寸相比較來進行該細胞增殖評估。The method of the third aspect of the invention preferably results in an inhibition of cell proliferation of the contacted cells. The phrase "inhibiting cell proliferation" is used to mean that a histone deacetylase inhibitor delays the growth of cells contacted with the inhibitor compared to untouched cells. Evaluation of cell proliferation was performed by counting contact and non-contact cells using a Coulter cell counter (Coulter, Miami, FL) or a hemocytometer. When the cells are in solid growth (eg, solid tumors or organs), the cell proliferation assessment can be performed by measuring growth with a caliper and comparing the contact cells to the growth size of the untouched cells.
與抑制劑接觸之細胞生長較佳與未接觸之細胞生長相比被延遲至少50%。更佳100%抑制細胞增殖(即經接觸之細胞數目不增加)。最佳短語"抑制細胞增殖"包括經接觸之細胞的數目或尺寸相較於未經接觸之細胞有所減少。因此，抑制所接觸之細胞中細胞增殖之本發明組蛋白去乙醯基酶抑制劑可誘發該經接觸之細胞遭受生長延遲、生長停滯、漸進式細胞死亡(即細胞凋零)或壞死性細胞死亡。The growth of cells in contact with the inhibitor is preferably delayed by at least 50% compared to the growth of untouched cells. More preferably, 100% inhibits cell proliferation (i.e., the number of cells that are contacted does not increase). The best phrase "inhibiting cell proliferation" involves a decrease in the number or size of cells contacted compared to untouched cells. Thus, the histone deacetylase inhibitor of the present invention which inhibits cell proliferation in the cells in contact can induce the contacted cells to suffer growth retardation, growth arrest, progressive cell death (i.e., cell dying) or necrotic cell death. .
本發明之組蛋白去乙醯基酶抑制劑之細胞增殖抑制能力使非同步生長細胞群體同步。舉例而言，本發明之組蛋白去乙醯基酶抑制劑可用於使活體外生長之非贅生性細胞群體停滯在細胞週期之G1或G2期。例如該同步允許識別在細胞週期G1或G2期過程中表現之基因及/或基因產物。培養細胞之該同步化亦可用於測試新轉染設計(protocol)之功效，其中細胞效力相異且視待轉染細胞之特殊細胞週期階段而定。使用本發明之組蛋白去乙醯基酶抑制劑使得細胞群體同步，藉此幫助偵測增強之轉染功效。The cell proliferation inhibiting ability of the histone deacetylase inhibitor of the present invention synchronizes the population of non-synchronized growth cells. For example, a histone deacetylase inhibitor of the invention can be used to arrest a population of non-neoplastic cells grown in vitro at the G1 or G2 phase of the cell cycle. For example, the synchronization allows for the identification of genes and/or gene products that are expressed during the G1 or G2 phase of the cell cycle. This synchronization of the cultured cells can also be used to test the efficacy of a new transfection protocol in which the cellular potency is different and depends on the particular cell cycle stage of the transfected cells. The use of the histone deacetylase inhibitors of the invention synchronizes the cell population, thereby helping to detect enhanced transfection efficiency.
一些較佳實施例中，該經接觸之細胞為一贅生性細胞。術語"贅生性細胞"用於表示一顯示異常細胞生長之細胞。較佳贅生性細胞之異常細胞生長為增加之細胞增長。一贅生性細胞可為增生性細胞，即顯示缺乏活體外生長接觸抑制性之細胞，一不能活體內轉移之良性腫瘤細胞，或能夠活體內轉移且試圖移除後會重現之癌細胞。術語"腫瘤發生"用於表示導致贅生性生長發展之細胞增殖誘發。一些實施例中，該組蛋白去乙醯基酶抑制劑誘發經接觸細胞中之細胞分化。因此，當與組蛋白去乙醯基酶抑制劑接觸時可誘發贅生性細胞分化，導致產生比經接觸之細胞系統發育上更先進的非贅生性子細胞。In some preferred embodiments, the contacted cells are a neoplastic cell. The term "neoplastic cells" is used to mean a cell that exhibits abnormal cell growth. Preferably, abnormal cell growth of the neoplastic cells is an increase in cell growth. A neoplastic cell can be a proliferating cell, that is, a cell that exhibits inhibition of in vitro growth contact inhibition, a benign tumor cell that cannot be transferred in vivo, or a cancer cell that can be transferred in vivo and that is reappeared after attempting to be removed. The term "tumorigenesis" is used to mean the induction of cell proliferation leading to the development of neoplastic growth. In some embodiments, the histone deacetylase inhibitor induces cell differentiation in the contacted cells. Thus, neoplastic cell differentiation can be induced when contacted with a histone deacetylase inhibitor, resulting in a more advanced non-neoplastic daughter cell than the contacted cell system.
在一些較佳實施例中該經接觸之細胞在動物體內。因此本發明提供一種用於治療動物體內細胞增殖性疾病或病症之方法，該方法包含向需要該治療之動物投與治療有效劑量之本發明組蛋白去乙醯基酶抑制劑。該動物較佳為一哺乳動物，更佳為馴養動物。該動物最佳為人類。In some preferred embodiments the contacted cells are in an animal. The invention therefore provides a method for treating a cell proliferative disease or disorder in an animal comprising administering to the animal in need of such treatment a therapeutically effective amount of a histone deacetylase inhibitor of the invention. Preferably, the animal is a mammal, more preferably a domesticated animal. The animal is best for humans.
術語"細胞增殖性疾病或病症"係指特徵為異常細胞生長、較佳為反常增加之細胞增殖的任何病症。該等細胞增殖性疾病或病症之實例包括(但不限於)癌症、再狹窄及牛皮癬。在尤其較佳實施例中，本發明提供一種用於抑制動物體內贅生性細胞增殖之方法，該方法包含向其體內具有至少一個贅生性細胞之動物投與治療有效劑量之本發明之組蛋白去乙醯基酶抑制劑。The term "cell proliferative disease or disorder" refers to any condition characterized by abnormal cell growth, preferably abnormally increased cell proliferation. Examples of such cell proliferative diseases or conditions include, but are not limited to, cancer, restenosis, and psoriasis. In a particularly preferred embodiment, the invention provides a method for inhibiting neoplastic cell proliferation in an animal, the method comprising administering to the animal having at least one neoplastic cell therein a therapeutically effective amount of a histone of the invention Inhibitor of acetylase.
預期本發明之一些化合物具有對來自原生動物源的組蛋白去乙醯基酶之抑制活性。因此，本發明亦提供一種用於治療或預防原生動物疾病或感染之方法，該方法包含向需要該治療之動物投與治療有效劑量之本發明之組蛋白去乙醯基酶抑制劑。該動物較佳為一哺乳動物，更佳為人類。較佳根據本發明之該實施例使用之組蛋白去乙醯基酶抑制劑抑制原生動物組蛋白去乙醯基酶之程度大於其抑制哺乳動物組蛋白去乙醯基酶(尤其為人類組蛋白去乙醯基酶)之程度。Some of the compounds of the invention are expected to have inhibitory activity against histone deacetylase from protozoa sources. Accordingly, the invention also provides a method for treating or preventing a protozoal disease or infection, the method comprising administering to the animal in need of such treatment a therapeutically effective amount of a histone deacetylase inhibitor of the invention. The animal is preferably a mammal, more preferably a human. Preferably, the histone deacetylase inhibitor used in this embodiment of the invention inhibits protozoal histone deacetylase more than its inhibitory mammalian histone deacetylase (especially human histone) To the extent of deacetylase).
本發明又提供一種用於治療真菌性疾病或感染之方法，該方法包含向需要該治療之動物投與治療有效劑量之本發明之組蛋白去乙醯基酶抑制劑。該動物較佳為哺乳動物，更佳為人類。較佳根據本發明此實施例使用之組蛋白去乙醯基酶抑制劑抑制真菌組蛋白去乙醯基酶之程度大於其抑制哺乳動物組蛋白去乙醯基酶(尤其為人類組蛋白去乙醯基酶)之程度。The invention further provides a method for treating a fungal disease or infection, the method comprising administering to the animal in need of such treatment a therapeutically effective amount of a histone deacetylase inhibitor of the invention. The animal is preferably a mammal, more preferably a human. Preferably, the histone deacetylase inhibitor used in this embodiment of the invention inhibits the fungal histone deacetylase to a greater extent than its inhibitory mammalian histone deacetylase (especially human histone to B) The degree of thiol enzyme).
術語"治療有效劑量"用以表示足以引起對受驗者細胞內組蛋白去乙醯基酶活性進行抑制作用的劑量，或足以抑制受驗者體內細胞增殖或誘發細胞分化之劑量。投藥可藉由任何途徑，包括(不限於)非經腸、口服、舌下、經皮、局部、鼻內、氣管內或直腸內。特定尤其較佳實施例中，在醫院環境中經靜脈內投與本發明之化合物。特定其它較佳實施例中較佳經口服途徑投藥。The term "therapeutically effective dose" is used to mean a dose sufficient to cause inhibition of histone deacetylase activity in a subject's cells, or a dose sufficient to inhibit cell proliferation or induce cell differentiation in a subject. Administration can be by any route including, without limitation, parenteral, oral, sublingual, transdermal, topical, intranasal, intratracheal or rectal. In a particularly particularly preferred embodiment, the compounds of the invention are administered intravenously in a hospital setting. In certain other preferred embodiments, it is preferred to administer the drug by the oral route.
全身投藥時，較佳投與組蛋白去乙醯基酶抑制劑之劑量足以達到約0.01μM至約100μM、更佳約0.05μM至約50μM、仍更佳約0.1μM至約25μM、又仍更佳約0.5μM至約25μM之抑制劑血液含量。對局部投藥而言比此濃度更低之濃度可為有效的，且可容忍更高之濃度。熟習此項技術者應瞭解產生治療效果所需之組蛋白去乙醯基酶抑制劑劑量可在相當程度上視組織、器官或待處理之特殊動物或患者而定變化。Preferably, when administered systemically, the dose of the histone deacetylase inhibitor is preferably from about 0.01 μM to about 100 μM, more preferably from about 0.05 μM to about 50 μM, still more preferably from about 0.1 μM to about 25 μM, still more Preferably, the blood content of the inhibitor is from about 0.5 μM to about 25 μM. Concentrations lower than this concentration for topical administration can be effective and can tolerate higher concentrations. Those skilled in the art will appreciate that the amount of histone deacetylase inhibitor required to produce a therapeutic effect will vary to a considerable extent depending on the tissue, organ or particular animal or patient being treated.
本發明第三態樣之特定較佳實施例中，該方法又包含使細胞與抑制組蛋白去乙醯基酶之表現的反義寡核苷酸接觸。核酸水平之抑制劑(例如反義寡核苷酸)與蛋白質水平之抑制劑(即組蛋白去乙醯基酶酶活性抑制劑)之組合導致改良之抑制效果，從而使為獲得給定抑制效果所需抑制劑量相較於其中任一者獨立使用時所需量而言有所減少。根據本發明此態樣之反義寡核苷酸互補於編碼HDAC－1、HDAC－2、HDAC－3、HDAC4、HDAC－5、HDAC－6、HDAC7及/或HDAC－8(例如參看關於HDAC－1之GenBank登錄號U50079，關於HDAC－2之GenBank登錄號U31814及關於HDAC－3之GenBank登錄號U75697)之RNA或雙股DNA區。In a particularly preferred embodiment of the third aspect of the invention, the method further comprises contacting the cells with an antisense oligonucleotide which inhibits the expression of histone deacetylase. The combination of an inhibitor of a nucleic acid level (eg, an antisense oligonucleotide) and a protein level inhibitor (ie, a histone deacetylase enzymatic activity inhibitor) results in an improved inhibitory effect, thereby achieving a given inhibitory effect The amount of inhibitor required is reduced compared to the amount required when either of them is used independently. An antisense oligonucleotide according to this aspect of the invention is complementary to encoding HDAC-1, HDAC-2, HDAC-3, HDAC4, HDAC-5, HDAC-6, HDAC7 and/or HDAC-8 (see, for example, HDAC) -1 of GenBank Accession No. U50079, GenBank Accession No. U31814 for HDAC-2 and GenBank Accession No. U75697 for HDAC-3, RNA or double strand DNA region.
為本發明之目的，術語"寡核苷酸"包括兩個或兩個以上脫氧核糖核苷、核糖核苷或2'－經取代之核糖核苷殘基或其任意組合之聚合物。該等寡核苷酸較佳具有約6至約100個核苷殘基，更佳約8至約50個核苷殘基，且最佳約12至約30個核苷殘基。該等核苷殘基可藉由眾多已知核苷間鍵之任一者彼此偶聯。該等核苷間鍵合包括(不限於)：硫代磷酸酯、二硫代磷酸酯、膦酸烷基酯、硫代膦酸烷基酯、磷酸三酯、胺基磷酸酯、矽氧烷、碳酸酯、羧甲酯、胺基乙酸酯(acetamidate)、胺基甲酸酯、硫醚、橋接胺基磷酸酯、橋接膦酸亞甲酯、橋接硫代磷酸酯及碸核苷間鍵。特定較佳實施例中，該等核苷間鍵可為磷酸二酯、磷酸三酯、硫代磷酸酯或胺基磷酸酯鍵或其組合。術語寡核苷酸亦容納該等具有經化學改質之鹼或糖及/或具有額外取代基之聚合物，包括(不限於)親脂性基、嵌入劑、二胺及金剛烷。For the purposes of the present invention, the term "oligonucleotide" includes polymers of two or more deoxyribonucleosides, ribonucleosides or 2'-substituted ribonucleoside residues or any combination thereof. Preferably, the oligonucleotides have from about 6 to about 100 nucleoside residues, more preferably from about 8 to about 50 nucleoside residues, and most preferably from about 12 to about 30 nucleoside residues. The nucleoside residues can be coupled to each other by any of a number of known internucleoside linkages. Such internucleoside linkages include, without limitation: phosphorothioates, dithiophosphates, alkyl phosphonates, alkyl thiophosphonates, phosphotriesters, amine phosphates, oxiranes , carbonate, carboxymethyl ester, acetamidate, urethane, thioether, bridged amino phosphate, bridged phosphonimide, bridged phosphorothioate and purine internucleoside linkage . In a particularly preferred embodiment, the internucleoside linkages can be phosphodiesters, phosphotriesters, phosphorothioates or amine phosphate linkages or combinations thereof. The term oligonucleotide also encompasses such polymers having chemically modified bases or sugars and/or having additional substituents including, without limitation, lipophilic groups, intercalators, diamines, and adamantane.
為本發明之目的，術語"2'－經取代之核糖核苷"包括其中戊糖部分2'位之羥基經取代產生2'－O－經取代之核糖核苷的核糖核苷。較佳該取代係以含有1－6個飽和或不飽和碳原子之低碳數烷基、或具有2－6個碳原子之芳基或烯丙基進行，其中該烷基、芳基或烯丙基可未經取代或經例如下述基團取代：鹵基、羥基、三氟甲基、氰基、硝基、醯基、醯氧基、烷氧基、羧基、烷氧羰基或胺基。術語"2'－經取代之核糖核苷"亦包括2'－羥基經胺基或較佳為氟之鹵基置換的核糖核苷。For the purposes of the present invention, the term "2'-substituted ribonucleoside" includes ribonucleosides in which the hydroxyl group at the 2' position of the pentose moiety is substituted to produce a 2'-O-substituted ribonucleoside. Preferably, the substitution is carried out with a lower alkyl group having 1 to 6 saturated or unsaturated carbon atoms or an aryl or allyl group having 2 to 6 carbon atoms, wherein the alkyl group, the aryl group or the alkene group The propyl group may be unsubstituted or substituted with, for example, a halogen group, a hydroxyl group, a trifluoromethyl group, a cyano group, a nitro group, a decyl group, a decyloxy group, an alkoxy group, a carboxyl group, an alkoxycarbonyl group or an amine group. . The term "2'-substituted ribonucleoside" also includes ribonucleosides in which the 2'-hydroxy group is substituted with an amine group or preferably a halo group of fluorine.
本發明此態樣利用之尤其較佳之反義寡核苷酸包括嵌合寡核苷酸及雜合寡核苷酸。Particularly preferred antisense oligonucleotides for use in this aspect of the invention include chimeric oligonucleotides and hybrid oligonucleotides.
為本發明之目的，"嵌合寡核苷酸"係指具有多於一種核苷間鍵之寡核苷酸。該嵌合寡核苷酸之一較佳實例為包含硫代磷酸酯、磷酸二酯或二硫代磷酸酯區之嵌合寡核苷酸，其較佳包含約2至約12個核苷酸，及膦酸烷基酯或硫代磷酸烷基酯區(例如參見Pederson等人美國專利第5,635,377號及第5,366,878號)。該等嵌合寡核苷酸較佳含有至少三個選自磷酸二酯及硫代磷酸酯鍵或其組合之連續核苷間鍵。For the purposes of the present invention, "chimeric oligonucleotide" refers to an oligonucleotide having more than one internucleoside linkage. A preferred example of one of the chimeric oligonucleotides is a chimeric oligonucleotide comprising a phosphorothioate, phosphodiester or phosphorodithioate region, preferably comprising from about 2 to about 12 nucleotides. And alkyl phosphonate or alkyl thiophosphate (see, for example, U.S. Patent Nos. 5,635,377 and 5,366,878 to Pederson et al.). Preferably, the chimeric oligonucleotides comprise at least three consecutive internucleoside linkages selected from the group consisting of phosphodiester and phosphorothioate linkages or combinations thereof.
為本發明之目的，"雜合寡核苷酸"係指具有多於一種核苷之寡核苷酸。該雜合寡核苷酸之一較佳實例包含核糖核苷酸或2'－經取代之核糖核苷酸區，其較佳包含約2至約12個2'－經取代之核苷酸，及脫氧核糖核苷酸區。較佳該雜合寡核苷酸含有至少三個連續脫氧核糖核苷且亦含有核糖核苷、2'－經取代之核糖核苷，較佳為2'－O－經取代之核糖核苷或其組合(例如參見Metelev及Agrawal，美國專利第5,652,355號)。For the purposes of the present invention, "hybrid oligonucleotide" refers to an oligonucleotide having more than one nucleoside. A preferred example of one of the hybrid oligonucleotides comprises a ribonucleotide or a 2'-substituted ribonucleotide region, which preferably comprises from about 2 to about 12 2'-substituted nucleotides, And a deoxyribonucleotide region. Preferably, the hybrid oligonucleotide comprises at least three consecutive deoxyribonucleosides and also contains a ribonucleoside, a 2'-substituted ribonucleoside, preferably a 2'-O-substituted ribonucleoside or Combinations thereof (see, for example, Metelev and Agrawal, U.S. Patent No. 5,652,355).
只要寡核苷酸保留其抑制所討論基因之表現的能力，本發明中利用之反義寡核苷酸之精確核苷酸序列及化學結構可不同。藉由測試特殊反義寡核苷酸是否具有活性很容易確定此點。針對此目的之可用檢定包括定量化編碼基因產物之mRNA、基因產物之西方墨點分析檢定、酶活性基因產物之活性檢定，或軟瓊脂生長檢定，或報導基因結構檢定，或活體內腫瘤生長檢定，上述所有者詳細描述於本說明書或Ramchandani等人(1997)Proc.Natl.Acad.Sci.USA 94：684－689中。The precise nucleotide sequence and chemical structure of the antisense oligonucleotides utilized in the present invention may vary as long as the oligonucleotide retains its ability to inhibit the expression of the gene in question. This is easily determined by testing whether a particular antisense oligonucleotide is active. Available assays for this purpose include quantification of mRNA encoding the gene product, western blot analysis of the gene product, activity assay of the enzyme active gene product, or soft agar growth assay, or reporting of gene structure assays, or in vivo tumor growth assays. The above-mentioned owner is described in detail in this specification or Ramchandani et al. (1997) Proc. Natl. Acad. Sci. USA 94:684-689.
利用熟知之化學方法可在適當固體載體上方便地合成出本發明中所用之反義寡核苷酸，該等方法包括H－膦酸酯化學，亞磷醯胺化學，或H－膦酸酯化學與亞磷醯胺化學的組合(換言之，某些循環週期使用H－膦酸酯化學，而其它循環週期則用亞磷醯胺化學)。適當固體載體包括任何用於固相寡核苷酸合成之標準固體載體，諸如可控孔徑玻璃(controlled－poreglass；CPG)(例如參見Pon,R.T.(1993)Methods in Molec.Bio1.20：465－496)。The antisense oligonucleotides used in the present invention can be conveniently synthesized on a suitable solid support by well-known chemical methods, including H-phosphonate chemistry, phosphorous amide chemistry, or H-phosphonate. The combination of chemistry and phosphorous amide chemistry (in other words, some cycles use H-phosphonate chemistry, while other cycles use phosphite chemistry). Suitable solid carriers include any standard solid support for solid phase oligonucleotide synthesis, such as controlled-poreglass (CPG) (see, for example, Pon, RT (1993) Methods in Molec. Bio 1.20:465- 496).
尤佳之寡核苷酸具有包括表A中所示核苷酸序列之約13至約35個核苷酸之核苷酸序列。另外仍尤佳之寡核苷酸具有約15至約26個表A所示核苷酸序列之核苷酸的核苷酸序列。A particularly preferred oligonucleotide has a nucleotide sequence comprising from about 13 to about 35 nucleotides of the nucleotide sequence shown in Table A. Still further preferred oligonucleotides have a nucleotide sequence of from about 15 to about 26 nucleotides of the nucleotide sequence shown in Table A.
以下實例旨在進一步說明本發明之某些較佳實施例，且不欲限制本發明之範疇。The following examples are intended to further illustrate certain preferred embodiments of the invention and are not intended to limit the scope of the invention.
N－(2－胺基－苯基)－3－[4－({(2－羥基－乙基)－[2－(1H－吲哚－3－基)－乙基]－胺基}－甲基)－苯基]－丙烯醯胺(6)步驟1：3－(4－甲醯基－苯基)－丙烯酸甲酯(1) 室溫下向1,2－二氯乙烷(100 mL)中之4－甲醯基肉桂酸(15.39 g，87.36 mmol)經攪拌懸浮液中分別加入濃硫酸(8 mL)及無水MeOH(15 mL)。將該反應混合物回流18小時，冷卻至室溫且濃縮之。以AcOEt稀釋該殘餘物且將其以H2 O、飽和NaHCO3 水溶液、H2 O及鹽水洗滌，經MgSO4 乾燥，過濾且再次濃縮。(溶離劑AcOEt/己烷：20/80→30/70)由急驟層析法經矽膠純化該粗產物得到呈淺黃色粉末之標題化合物2(9.75 g，51.26 mmol，產率59%)。1 H NMR(300 MHz,CDCl3 )δ(ppm)：10.04(s,1H),7.91(d,J＝7.9 Hz,2H),7.80－7.60(m,3H),6.56(d,J＝15.8 Hz,1H),3.84(s,3H)。 N-(2-Amino-phenyl)-3-[4-({(2-hydroxy-ethyl)-[2-(1H-indol-3-yl)-ethyl]-amino}- Methyl)-phenyl]-acrylamide (6) Step 1: 3-(4-Methyl-phenyl-phenyl)-methyl acrylate (1) to 1,2-dichloroethane at room temperature (100) 4-Methyl cinnamic acid (15.39 g, 87.36 mmol) in mL) was added with concentrated sulphuric acid (8 mL) and anhydrous MeOH (15 mL). The reaction mixture was refluxed for 18 h, cooled to rt and concentrated. The residue was diluted with AcOEt and dried which was washed with H 2 O, saturated aqueous NaHCO 3, H 2 O and brine and over MgSO 4, filtered and concentrated again. The lysate (AcOEt/hexane: 20/80, 30/70). 1 H NMR (300 MHz, CDCl 3 ) δ (ppm): 10.04 (s, 1H), 7.91 (d, J = 7.9 Hz, 2H), 7.80-7.60 (m, 3H), 6.56 (d, J = 15.8) Hz, 1H), 3.84 (s, 3H).
步驟2：3－(4－{[2－(1H－吲哚－3－基)－乙胺]－甲基}－苯基)－丙烯酸甲酯(2)在室溫氮氣下向溶於無水1,2－二氯乙烷(200 mL)中之1(3.00 g，15.77 mmol)及色胺(2.78 g，17.35 mmol)在經攪拌溶液中加入NaBH(OAc)3 (3.87 g，17.35 mmol)。室溫下攪拌反應混合物39小時，將其傾入至10% K2 CO3 溶液且以CH2 Cl2 萃取之。濃縮有機層形成殘餘物，其(MeOH/CH2 Cl2 ：10/90)由急驟層析法經矽膠純化且在AcOEt/己烷之混合物中共沉澱產生呈黃色固體之標題化合物2(4.39 g，13.13 mmol，含量83%)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：10.78(s,1H),7.70－7.62(m,3H),7.49(d,J＝8.0 Hz,1H),7.39(d,J＝8.2 Hz,2H),7.33(dt,J＝8.0,0.9 Hz,1H),7.13(d,J＝2.2 Hz,1H),7.06(ddd,J＝7.0,7.0,1.2 Hz,1H),6.96(ddd,J＝6.9,6.9,1.1 Hz,1H),6.62(d,J＝16.0 Hz,1H),3.79(s,2H),3.75(s,3H),2.91－2.78(m,4H),2.18(bs,1H)。Step 2: 3-(4-{[2-(1H-indol-3-yl)-ethylamine]-methyl}-phenyl)-methyl acrylate (2) is dissolved in anhydrous at room temperature under nitrogen 1 (3.00 g, 15.77 mmol) of 1,2-dichloroethane (200 mL) and tryptamine (2.78 g, 17.35 mmol) were added NaBH(OAc) 3 (3.87 g, 17.35 mmol) to a stirred solution. . The reaction mixture was stirred at room temperature for 39 hours, it was poured into 10% K 2 CO 3 solution and extracted with CH 2 Cl 2 to it. The organic layer was concentrated and the residue was formed, which (MeOH / CH 2 Cl 2: 10/90) was purified by flash chromatography over silica gel and precipitate the title compound as a yellow solid of 2 (4.39 g in AcOEt / hexane mixtures of CCP 13.13 mmol, content 83%). 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 10.78 (s, 1H), 7.70-7.62 (m, 3H), 7.49 (d, J = 8.0 Hz, 1H), 7.39 (d, J) = 8.2 Hz, 2H), 7.33 (dt, J = 8.0, 0.9 Hz, 1H), 7.13 (d, J = 2.2 Hz, 1H), 7.06 (ddd, J = 7.0, 7.0, 1.2 Hz, 1H), 6.96 (ddd, J=6.9, 6.9, 1.1 Hz, 1H), 6.62 (d, J = 16.0 Hz, 1H), 3.79 (s, 2H), 3.75 (s, 3H), 2.91-2.78 (m, 4H), 2.18 (bs, 1H).
步驟3：3－[4－({[2－(第三丁基－二甲基－矽烷氧基)－乙基]－[2－(1H－吲哚－3－基)－乙基]－胺基}－甲基)－苯基]－丙烯酸甲酯(3) 在室溫氮氣下向溶於DMSO(22 mL)中之2(2.82 g，8.44 mmol)及二異丙基乙胺(2.21 mL，12.66 mmol)經攪拌溶液中加入(2－溴－乙氧基)－第三丁基－二甲基矽烷(2.17 mL，10.12 mmol)。在50－55℃下加熱該反應混合物24小時，將其傾入水中且以CH2 Cl2 萃取。將該有機層經MgSO4 乾燥，過濾及濃縮。由急驟層析法經矽膠(AcOEt/CH2 Cl2 ，15/85，加上少數滴NH4 OH)上純化該粗產物產生深橙色油狀標題化合物3(4.06 g，8.24 mmol，產率97%)。1 H NMR(300 MHz,CDCl3 )δ(ppm)：7.95(bs,1H)，7.70(d,J＝15.8 Hz,1H),7.58－7.30(m,6H),7.18(t,J＝7.5 Hz,1H),7.07(t,J＝7.5 Hz,1H),7.00(bs,1H),6.43(d,J＝16.2 Hz,1H),3.88－3.68(m,7H),3.04－2.66(m,6H),0.88(bs,9H),0.04(bs,6H)。 Step 3: 3-[4-({[2-(T-butyl-dimethyl-decyloxy)-ethyl]-[2-(1H-indol-3-yl)-ethyl]- Amino}-methyl)-phenyl]-methyl acrylate (3) 2 (2.82 g, 8.44 mmol) and diisopropylethylamine (2.21) dissolved in DMSO (22 mL) at room temperature under nitrogen. (2-bromo-ethoxy)-tert-butyl-dimethyldecane (2.17 mL, 10.12 mmol) was added to a stirred solution. The reaction was heated at 50-55 deg.] C the mixture for 24 hours, poured into water and extracted in CH 2 Cl 2. The organic layer was dried over MgSO 4, filtered and concentrated. By flash chromatography over silica gel (AcOEt / CH 2 Cl 2, 15/85, plus a few drops of NH 4 OH) of the crude product was purified on a dark orange oil of the title compound 3 (4.06 g, 8.24 mmol, yield 97 %). 1 H NMR (300 MHz, CDCl 3 ) δ (ppm): 7.95 (bs, 1H), 7.70 (d, J = 15.8 Hz, 1H), 7.58-7.30 (m, 6H), 7.18 (t, J = 7.5) Hz, 1H), 7.07 (t, J = 7.5 Hz, 1H), 7.00 (bs, 1H), 6.43 (d, J = 16.2 Hz, 1H), 3.88-3.68 (m, 7H), 3.04-2.66 (m) , 6H), 0.88 (bs, 9H), 0.04 (bs, 6H).
步驟4：3－[4－({[2－(第三丁基－二甲基－矽烷氧基)－乙基]－[2－(1H－吲哚－3－基)－乙基]－胺基}－甲基)－苯基]－丙烯酸(4) 在室溫下向THF(40 mL)中之化合物3(3.18 g，6.45 mmol)經攪拌溶液中加入LiOH.H2 O(677 mg，16.14 mmol)之水(20 mL)溶液。24小時後將該反應混合物濃縮，以水稀釋且用1N HCl酸化至pH為5－6。將形成之沉澱物經過濾分離，以水沖洗及乾燥產生呈乳白色固體之標題化合物4(2.43 g，5.08 mmol，產率79%)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：12.34(bs,1H),10.75(s,1H),7.63(d,J＝8.2 Hz,2H),7.59(d,J＝15.8 Hz,1H),7.44－7.35(m,3H),7.32(d,J＝8.0 Hz,1H),7.11(d,J＝2.3 Hz,1H),7.05(td,J＝7.5,1.0 Hz,1H),6.92(td,J＝7.4,0.9 Hz,1H),6.51(d,J＝15.8 Hz,1H),3.79(s,2H),3.69(t,J＝6.4 Hz,2H),2.93－2.74(m,4H),2.69(t,J＝6.2 Hz,2H),0.88(s,9H),0.05(s,6H)。 Step 4: 3-[4-({[2-(T-butyl-dimethyl-decyloxy)-ethyl]-[2-(1H-indol-3-yl)-ethyl]- Amino}-methyl)-phenyl]-acrylic acid (4) To a solution of compound 3 (3.18 g, 6.45 mmol) in THF (40 mL ) at room temperature was added LiOH.H 2 O (677 mg) , 16.14 mmol) of water (20 mL) solution. After 24 hours the reaction mixture was concentrated, diluted with water and EtOAc EtOAc EtOAc. The resulting precipitate was isolated by EtOAc (EtOAc m. 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 12.34 (bs, 1H), 10.75 (s, 1H), 7.63 (d, J = 8.2 Hz, 2H), 7.59 (d, J = 15.8) Hz,1H),7.44-7.35(m,3H),7.32(d,J=8.0 Hz,1H),7.11(d,J=2.3 Hz,1H),7.05(td,J=7.5,1.0 Hz,1H ), 6.92 (td, J = 7.4, 0.9 Hz, 1H), 6.51 (d, J = 15.8 Hz, 1H), 3.79 (s, 2H), 3.69 (t, J = 6.4 Hz, 2H), 2.93 - 2.74 (m, 4H), 2.69 (t, J = 6.2 Hz, 2H), 0.88 (s, 9H), 0.05 (s, 6H).
步驟5：N－(2－胺基－苯基)－3－[4－({[2－(第三丁基－二甲基－矽烷氧基)乙基]－[2－(1H－吲哚－3－基)－乙基]－胺基}－甲基)－苯基]－丙烯醯胺(5)在室溫氮氣下向溶於無水DMF(20 mL)中之4(1.30 g，2.72 mmol)的經攪拌溶液中分別加入Et3 N(330μl，3.26 mmol)及BOP試劑(1.32 g，2.99 mmol)。30分鐘後，逐滴添加1,2－苯二胺(352 mg，3.26 mmol)、Et3 N(1.14 mL，8.15 mmol)之無水DMF(3 mL)溶液。3小時後將反應混合物傾入至NH4 Cl飽和水溶液中，且以AcOEt萃取之。將該萃取物以飽和NH4 Cl、水及鹽水洗滌，經MgSO4 乾燥，將其過濾、濃縮且由急驟層析法經矽膠(MeOH/CH2 Cl2 ，5/95加上若干滴NH4 OH)純化產生黃色黏稠發泡體狀標題化合物5(1.49 g，2.62 mmol，產率96%)。1 H NMR(300 MHz,DMSO－d6 )δ(ppm)：10.78(s,1H),9.40(s,1H),7.59(d,J＝8.0 Hz,2H),7.58(d,J＝15.8 Hz,1H),7.45(d,J＝7.9 Hz,2H),7.40(t,J＝7.7 Hz,2H),7.35(d,J＝8.4 Hz,1H),7.14(s,1H),7.07(t,J＝7.5 Hz,1H),7.05－6.85(m,3H),6.79(d,J＝7.9 Hz,1H),6.62(t,J＝7.5 Hz,1H),4.98(bs,2H),3.80(s,2H),3.71(t,J＝6.2 Hz,2H),2.95－2.75(m,4H),2.71(t,J＝6.2 Hz,2H),0.89(s,9H),0.05(s,6H)。Step 5: N-(2-Amino-phenyl)-3-[4-({[2-(t-butyl-dimethyl-decyloxy)ethyl]-[2-(1H-吲)哚-3-yl)-ethyl]-amino}-methyl)-phenyl]-propenylamine (5) 4 (1.30 g, dissolved in anhydrous DMF (20 mL) at room temperature under nitrogen. 2.72 mmol) a stirred solution were added Et 3 N (330μl, 3.26 mmol ) and BOP reagent (1.32 g, 2.99 mmol). After 30 minutes, was added dropwise 1,2-phenylenediamine (352 mg, 3.26 mmol), Et 3 N (1.14 mL, 8.15 mmol) of anhydrous DMF (3 mL) was added. After 3 hours, the reaction mixture was poured into a saturated aqueous solution of NH 4 Cl and extracted with AcOEt. Drying the extract was washed with saturated NH 4 CI, water and brine over MgSO 4, filtered, and concentrated over silica gel (MeOH / CH 2 Cl 2, 5/95 plus several drops by flash chromatography NH 4 The title compound 5 (1.49 g, 2.62 mmol, yield 96%) 1 H NMR (300 MHz, DMSO-d 6 ) δ (ppm): 10.78 (s, 1H), 9.40 (s, 1H), 7.59 (d, J = 8.0 Hz, 2H), 7.58 (d, J = 15.8) Hz, 1H), 7.45 (d, J = 7.9 Hz, 2H), 7.40 (t, J = 7.7 Hz, 2H), 7.35 (d, J = 8.4 Hz, 1H), 7.14 (s, 1H), 7.07 ( t, J = 7.5 Hz, 1H), 7.05-6.85 (m, 3H), 6.79 (d, J = 7.9 Hz, 1H), 6.62 (t, J = 7.5 Hz, 1H), 4.98 (bs, 2H), 3.80(s, 2H), 3.71 (t, J = 6.2 Hz, 2H), 2.95-2.75 (m, 4H), 2.71 (t, J = 6.2 Hz, 2H), 0.89 (s, 9H), 0.05 (s) , 6H).
步驟6：N－(2－胺基－苯基)－3－[4－({(2－羥基－乙基)－[2－(1H－吲哚－3－基)－乙基]－胺基}－甲基)－苯基]－丙烯醯胺(6) 在－20℃下於氮氣下向5(1.49 g，2.62 mmol)之經攪拌無水THF(30 mL)溶液中緩緩加入TBAF(2.88 mL，2.88 mmol，THF中1.0 M)。將該反應混合物經1小時溫至室溫且再攪拌22小時。添加MeOH且濃縮反應混合物，將其以AcOEt稀釋，且相繼以NaHCO3 飽和水溶液、H2 O、NH4 Cl飽和水溶液及鹽水洗滌，經MgSO4 乾燥之，過濾且加以濃縮。由急驟層析法經矽膠(MeOH/CH2 Cl2 ，5/95→10/90加上若干滴NH4 OH)純化殘餘物且以AcOEt/CH2 Cl2 /己烷之混合物濕磨產生呈淺黃色固體之標題化合物6(956 mg，2.10 mmol，產率80%)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：10.76(s,1H),9.39(s,1H),AB系統(δA ＝7.58,δB ＝7.44,JA B ＝8.0 Hz,4H),7.56(d,J＝15.7 Hz,1H),7.42－7.34(m,2H),7.33(d,J＝8.0 Hz,1H),7.12(d,J＝2.3 Hz,1H),7.05(td,J＝7.2,1.2 Hz,1H),6.98－6.90(m,2H),6.90(d,J＝15.8 Hz,1H),6.77(dd,J＝8.0,1.4 Hz,1H),6.60(ddd,J＝7.5,7.5,1.4 Hz,1H),4.98(bs,2H),4.43(t,J＝5.4 Hz,1H),3.78(s,2H),3.56(td,J＝6.3,5.6 Hz,2H),2.94－2.84(m,2H),2.82－2.74(m,2H),2.68(t,J＝6.5 Hz,2H)。 Step 6: N-(2-Amino-phenyl)-3-[4-({(2-hydroxy-ethyl)-[2-(1H-indol-3-yl)-ethyl]-amine } - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ( ) - ( - - - - - - 2.88 mL, 2.88 mmol, 1.0 M in THF. The reaction mixture was warmed to room temperature over 1 h and stirred for additional 22 h. MeOH was added and the reaction mixture was concentrated, it was diluted with AcOEt, H 2 O and washed sequentially with saturated aqueous NaHCO 3,, NH 4 Cl saturated solution and brine, dried over MgSO 4, filtered and concentrated. Was purified by flash chromatography over silica gel (MeOH / CH 2 Cl 2, 5/95 → 10/90 plus a few drops of NH 4 OH) and the residue was purified in AcOEt / CH mixture 2 Cl 2 / hexane to produce the triturated The title compound 6 (956 mg, 2.10 mmol, yield 80%) 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 10.76 (s, 1H), 9.39 (s, 1H), AB system (δ A = 7.58, δ B = 7.44, J A B = 8.0 Hz , 4H), 7.56 (d, J = 15.7 Hz, 1H), 7.42 - 7.34 (m, 2H), 7.33 (d, J = 8.0 Hz, 1H), 7.12 (d, J = 2.3 Hz, 1H), 7.05 (td, J = 7.2, 1.2 Hz, 1H), 6.98-6.90 (m, 2H), 6.90 (d, J = 15.8 Hz, 1H), 6.77 (dd, J = 8.0, 1.4 Hz, 1H), 6.60 ( Ddd, J=7.5, 7.5, 1.4 Hz, 1H), 4.98 (bs, 2H), 4.43 (t, J = 5.4 Hz, 1H), 3.78 (s, 2H), 3.56 (td, J = 6.3, 5.6 Hz , 2H), 2.94-2.84 (m, 2H), 2.82-2.74 (m, 2H), 2.68 (t, J = 6.5 Hz, 2H).
N－(2－胺基－苯基)－4－({4－[2－(3,4－二甲氧基－苯基)－乙胺基]－嘧啶－2－基胺基}－甲基)－苯甲醯胺(10a)步驟1：(2－氯－嘧啶－4－基)－[2－(3,4－二甲氧基－苯基)－乙基]－胺(7) 在室溫下氮氣下向溶於無水DMF(10 mL)中之2,4－二氯嘧啶(500 mg，3.36 mmol)之經攪拌溶液中分別緩緩加入i－Pr2 NEt(1.06 mL，6.10 mmol)及3,4－二甲氧基苯乙胺(531μl，3.05 mmol)。24小時後以AcOEt稀釋反應混合物且將其相繼以NH4 Cl飽和水溶液及鹽水洗滌，經MgSO4 乾燥之，過濾且加以濃縮。由急驟層析法經矽膠(MeOH/CH2 Cl2 ：2/98→5/95)純化殘餘物產生呈淺黃色油狀物之標題化合物7a(744 mg，2.53 mmol，產率83%)。 N-(2-Amino-phenyl)-4-({4-[2-(3,4-dimethoxy-phenyl)-ethylamino]-pyrimidin-2-ylamino}-- Benzo-benzamide (10a) Step 1: (2-Chloro-pyrimidin-4-yl)-[2-(3,4-dimethoxy-phenyl)-ethyl]-amine (7) i-Pr 2 NEt (1.06 mL, 6.10) was slowly added to the stirred solution of 2,4-dichloropyrimidine (500 mg, 3.36 mmol) dissolved in anhydrous DMF (10 mL) at room temperature under nitrogen. Methyl) and 3,4-dimethoxyphenethylamine (531 μl, 3.05 mmol). After 24 hours the reaction mixture was diluted with AcOEt and washed sequentially with which saturated NH 4 Cl solution and brine, dried over MgSO 4, filtered and concentrated. By flash chromatography over silica gel (MeOH / CH 2 Cl 2: 2/98 → 5/95) as a pale yellow residue was purified oil of the title compound 7a (744 mg, 2.53 mmol, 83% yield).
步驟2：4－({4－[2－(3,4－二甲氧基－苯基)－乙胺基]－嘧啶－2－基胺基}－甲基)－苯甲酸甲酯(8) 在一密閉燒瓶中，將異丙醇(50 mL)中之7(744 mg，2.53 mmol)、4－(胺甲基)苯甲酸甲酯(628 mg，3.80 mmol)及i－Pr2 NEt(882μl，5.07 mmol)混合物加熱至120－125℃歷時7天(在此期間向反應混合物中加入過量4－(胺甲基)苯甲酸甲酯)。使反應混合物冷卻至室溫，濃縮且由急驟層析法經矽膠(MeOH/CH2 Cl2 ，5/95→10/90加上若干滴NH4 OH)純化以產生呈橙色黏稠固體之標題化合物8(671 mg，1.59 mmol，產率63%)。 Step 2: 4-({4-[2-(3,4-Dimethoxy-phenyl)-ethylamino]-pyrimidin-2-ylamino}-methyl)-benzoic acid methyl ester (8 7 (744 mg, 2.53 mmol), 4-(aminomethyl)benzoic acid methyl ester (628 mg, 3.80 mmol) and i-Pr 2 NEt in isopropanol (50 mL ) in a closed flask (882 μl, 5.07 mmol) The mixture was heated to 120-125 ° C for 7 days (in the meantime, an excess of methyl 4-(aminomethyl)benzoate) was added to the reaction mixture. The reaction mixture was cooled to rt, concentrated and purified by flash chromatography over silica gel (MeOH / CH 2 Cl 2, 5/95 → 10/90 plus a few drops of NH 4 OH) to yield purified sticky orange solid of the title compound 8 (671 mg, 1.59 mmol, yield 63%).
步驟3：4－((4－[2－(3,4－二甲氧基－苯基)－乙胺基]－嘧啶－2－基胺基)－甲基)－苯甲酸(9) 在室溫下向THF(15 mL)中之化合物8(670 mg，1.59 mmol)的經攪拌溶液中加入LiOH.H2 O(166 mg，3.97 mmol)之水(5 mL)溶液。24小時後濃縮反應混合物、將其以水稀釋且以2 N HCl(pH為5－6)酸化之。形成沉澱物，其經過濾分離，以水沖洗且經乾燥產生呈乳白色固體之標題化合物9(600 mg，1.47 mmol，產率93%)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：AB系統(δA ＝7.87,δB ＝7.41,J＝8.2 Hz,4H),7.68－7.58(m,1H),7.12－6.56(m,5H),5.75(d,J＝5.5 Hz,1H),4.53(d,J＝6.3 Hz,2H),3.74及3.72(2s,6H),3.48－3.30(m,2H),2.80－2.60(m,2H)。 Step 3: 4-((4-[2-(3,4-Dimethoxy-phenyl)-ethylamino]-pyrimidin-2-ylamino)-methyl)-benzoic acid (9) A solution of LiOH.H 2 O (166 mg, 3.97 mmol) in water (5 mL) was added to a stirred solution of compound 8 (670 mg, 1.59 mmol) in THF (15 mL). After 24 hours, the reaction mixture was concentrated, diluted with water and acidified with 2 N HCl (pH 5-6). Precipitate was formed which was isolated by filtration, washed with water and dried to give the title compound 9 (600 mg, 1.47 mmol, yield 93%). 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): AB system (δ A = 7.87, δ B = 7.41, J = 8.2 Hz, 4H), 7.68-7.58 (m, 1H), 7.12-6.56 (m, 5H), 5.75 (d, J = 5.5 Hz, 1H), 4.53 (d, J = 6.3 Hz, 2H), 3.74 and 3.72 (2s, 6H), 3.48-3.30 (m, 2H), 2.80- 2.60 (m, 2H).
步驟4：N－(2－胺基－苯基)4－({4－[2－(3,4－二甲氧基－苯基)－乙胺基]－嘧啶－2－基胺基}－甲基)－苯甲醯胺(10a) 在室溫下氮氣下向溶於無水DMF(10 mL)中之9a(300 mg，0.73 mmol)的經攪拌溶液中分別加入Et3 N(123μl，0.88 mmol)及BOP試劑(358 mg，0.81 mmol)。30分鐘後，逐滴添加1,2－苯二胺(95 mg，0.88 mmol)，Et3 N(307μl，2.20 mmol)之無水DMF(2 mL)溶液。攪拌隔夜後，將反應混合物傾入至NH4 Cl飽和水溶液中且以AcOEt萃取之。相繼以飽和NH4 Cl、水及鹽水洗滌有機層，將其經MgSO4 乾燥，過濾且濃縮之。由急驟層析法經矽膠(MeOH/CH2 Cl2 ：5/95→10/90加上少數滴NH4 OH)純化該殘餘物且在AcOEt/MeOH/己烷之混合物中共沉澱產生呈乳白色固體之標題化合物10a(280 mg，0.56 mmol，產率76%)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.60(s,1H),AB系統(δA ＝7.91,δB ＝7.43,J＝8.0 Hz,4H),7.71－7.58(m,1H),7.17(d,J＝7.4 Hz,1H),7.20－7.00(m,2H),6.98(t,J＝7.5 Hz,1H),6.86(d,J＝8.0 Hz,1H),6.84－6.64(m,3H),6.61(t,J＝7.4 Hz,1H),5.76(d,J＝5.3 Hz,1H),4.90(bs,2H),4.54(d,J＝6.1 Hz,2H),3.74(s,6H),3.50－3.35(m,2H),2.80－2.62(m,2H)。 Step 4: N-(2-Amino-phenyl)4-({4-[2-(3,4-dimethoxy-phenyl)-ethylamino]-pyrimidin-2-ylamino} - methyl) - benzoyl amine (10a) of anhydrous 9a (10 mL), dissolved in DMF at the room temperature under nitrogen (300 mg, 0.73 mmol) was added via respectively Et 3 N (123μl, 0.88 mmol) and BOP reagent (358 mg, 0.81 mmol). After 30 minutes, was added dropwise 1,2-phenylenediamine (95 mg, 0.88 mmol), Et 3 N (307μl, 2.20 mmol) of anhydrous DMF (2 mL) was added. After stirring overnight, the reaction mixture was poured into a saturated aqueous solution of NH 4 Cl and extracted with AcOEt. Successively, dried with saturated NH 4 Cl, water, the organic layer was washed with brine and over MgSO 4, filtered and concentrated. By flash chromatography over silica gel: The residue was purified (MeOH / CH 2 Cl 2 5 /95 → 10/90 plus a few drops of NH 4 OH) and the solid precipitate was milky white mixture AcOEt / MeOH / hexanes of CCP The title compound 10a (280 mg, 0.56 mmol, yield 76%). 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.60 (s, 1H), AB system (δ A = 7.91, δ B = 7.43, J = 8.0 Hz, 4H), 7.71-7.58 (m) , 1H), 7.17 (d, J = 7.4 Hz, 1H), 7.20-7.00 (m, 2H), 6.98 (t, J = 7.5 Hz, 1H), 6.86 (d, J = 8.0 Hz, 1H), 6.84 -6.64 (m, 3H), 6.61 (t, J = 7.4 Hz, 1H), 5.76 (d, J = 5.3 Hz, 1H), 4.90 (bs, 2H), 4.54 (d, J = 6.1 Hz, 2H) , 3.74 (s, 6H), 3.50-3.35 (m, 2H), 2.80-2.62 (m, 2H).
用與上文所述製備化合物10a之實例2(流程2)相同之程序製備實例3－11(化合物10b－10j)。Example 3-11 (Compounds 10b-10j) was prepared by the same procedure as Example 2 (Scheme 2) for the preparation of compound 10a described above.
N－(2－胺基－苯基)－4－([2,3']聯吡啶基－6－基胺甲基)－苯甲醯胺(13a)步驟1：[2,3']聯吡啶基－6－基胺(11) 在室溫下向2－胺基－6－嗅吡啶(5.38 g，31.09 mmol)、2,4,6－(3－吡啶基)－環三硼氧烷(3.80 g，12.07 mmol)及Na2 CO3 水溶液(100 mL，0.4 M)混合物的經攪拌脫氣乙腈(100 mL)懸浮液中加入Pd(PPh3 )4 (1.70 g，1.47 mmol)。在氮氣下在95℃下加熱該反應混合物1至2天，將其冷卻至室溫且過濾。由急驟層析法經矽膠(MeOH/CH2 Cl2 ：5/95→10/90加入幾滴NH4 OH)濃縮純化濾液且與AcOEt/CH2 Cl2 /己烷之混合物共沉澱產生呈淺黃色固體之標題化合物11(4.091 g，23.90 mmol，產率77%)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.16(dd,J＝2.2,0.8 Hz,1H),8.57(dd,J＝4.7,1.6 Hz 1H),8.33－8.28(m,1H),7.54－7.44(m,2H),7.14(dd,J＝7.3,0.5 Hz,1H),6.49(dd,J＝8.2,0.4 Hz 1H),6.12(bs,2H)。 N-(2-Amino-phenyl)-4-([2,3']bipyridyl-6-ylaminomethyl)-benzamide (13a) Step 1: [2,3'] Pyridyl-6-ylamine (11) to 2-amino-6-oleragylpyridine (5.38 g, 31.09 mmol), 2,4,6-(3-pyridyl)-cyclotriboroxane at room temperature (3.80 g, 12.07 mmol) and Na 2 CO 3 solution (100 mL, 0.4 M) stirred degassed mixture of acetonitrile (100 mL) was added Pd (PPh 3) 4 (1.70 g, 1.47 mmol). The reaction mixture was heated at 95 ° C for 1 to 2 days under nitrogen, cooled to room temperature and filtered. The filtrate was concentrated by flash chromatography (MeOH/CH 2 Cl 2 : 5/95→10/90, a few drops of NH 4 OH) and coprecipitated with a mixture of AcOEt/CH 2 Cl 2 /hexane to give a shallow The title compound 11 (4.091 g, 23.90 mmol, yield 77%) 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.16 (dd, J = 2.2, 0.8 Hz, 1H), 8.57 (dd, J = 4.7, 1.6 Hz 1H), 8.33-8.28 (m, 1H), 7.54-7.44 (m, 2H), 7.14 (dd, J = 7.3, 0.5 Hz, 1H), 6.49 (dd, J = 8.2, 0.4 Hz 1H), 6.12 (bs, 2H).
步驟2：4－([2,3']聯吡啶基－6－基胺甲基)－苯甲酸甲酯(12) 在室溫下向11(3.00 g，17.52 mmol)、4－甲醯基苯甲酸甲酯(4.62 g，28.11 mmo1，1.5－2.0等價)及氣化二丁基錫160 mg，0.53 mmol)之混合物的經攪拌無水THF(15 mL)懸浮液中經兩天w三份添加苯基矽烷(2.34 mL，19.28 mmol)。攪拌2至7天後過濾反應混合物，將濾液濃縮且由急驟層析法經矽膠(MeOH/CH2 Cl2 ，2/98→10/90)上純化產生呈淺黃色固體之標題化合物12(5.50 g，17.22 mmol，產率98%)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.11(dd,J＝2.3,0.7 Hz,1H),8.55(dd,J＝4.7,1.8 Hz 1H),8.29－8.24(m,1H),7.93(d,J＝8.4 Hz,2H),7.57－7.40(m,5H),7.18(d,J＝7.2 Hz,1H),6.59(d,J＝8.2 Hz 1H),4.69(d,J＝6.l Hz,2H),3.85(s,3H)。 Step 2: 4-([2,3']bipyridyl-6-ylaminomethyl)-benzoic acid methyl ester (12) at room temperature to 11 (3.00 g, 17.52 mmol), 4-carbazinyl a mixture of methyl benzoate (4.62 g, 28.11 mmo1, 1.5-2.0 equivalent) and dibutyltin vapor 160 mg, 0.53 mmol) in a stirred anhydrous THF (15 mL) suspension over two days w benzene Base decane (2.34 mL, 19.28 mmol). The title was stirred 2-7 days the reaction mixture was filtered and the filtrate was concentrated and purified by flash chromatography on a pale yellow solid by silica gel (MeOH / CH 2 Cl 2, 2/98 → 10/90) The compound 12 (5.50 g, 17.22 mmol, yield 98%). 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.11 (dd, J = 2.3, 0.7 Hz, 1H), 8.55 (dd, J = 4.7, 1.8 Hz 1H), 8.29-8.24 (m, 1H), 7.93 (d, J = 8.4 Hz, 2H), 7.57-7.40 (m, 5H), 7.18 (d, J = 7.2 Hz, 1H), 6.59 (d, J = 8.2 Hz 1H), 4.69 (d , J = 6.l Hz, 2H), 3.85 (s, 3H).
步驟3：N－(2－胺基－苯基)4－([2,3']聯吡啶基－6－基胺甲基)－苯甲醯胺(13a) 依照與實例2中相同之程序(流程2)自12以步驟3及4兩步獲得呈乳白色固體之標題化合物13a(實例2)。1 HNMR(400MHz,DMSO－d6 )δ(ppm)：9.60(s,1H),9.16(dd,J＝2.2,0.9 Hz,1H),8.56(dd,J＝4.8,1.7 Hz 1H),8.31(ddd,J＝7.8,2.3,1.7 Hz,1H),7.95(d,J＝8.2 Hz,2H),7.57－7.48(m,3H),7.46(ddd,J＝8.0,4.7,0.8 Hz,1H),7.42(t,J＝6.1 Hz,1H),7.19(dd,J＝7.2,0.6 Hz,1H),7.17(dd,J＝7.3,1.0 Hz,1H),6.98(td,J＝7.5,1.4,1H),6.79(dd,J＝7.8,1.4 Hz 1H),6.65－6.57(m,2H),4.90(bs,2H),4.69(d,J＝6.1 Hz,2H)。 Step 3: N-(2-Amino-phenyl)4-([2,3']bipyridinyl-6-ylaminomethyl)-benzimidamide (13a) according to the same procedure as in Example 2. (Scheme 2) The title compound 13a (Example 2) was obtained as a milky white solid from step 12 and step. 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.60 (s, 1H), 9.16 (dd, J = 2.2, 0.9 Hz, 1H), 8.56 (dd, J = 4.8, 1.7 Hz 1H), 8.31 (ddd, J = 7.8, 2.3, 1.7 Hz, 1H), 7.95 (d, J = 8.2 Hz, 2H), 7.57-7.48 (m, 3H), 7.46 (ddd, J = 8.0, 4.7, 0.8 Hz, 1H) ), 7.42 (t, J = 6.1 Hz, 1H), 7.19 (dd, J = 7.2, 0.6 Hz, 1H), 7.17 (dd, J = 7.3, 1.0 Hz, 1H), 6.98 (td, J = 7.5, 1.4, 1H), 6.79 (dd, J = 7.8, 1.4 Hz 1H), 6.65-6.57 (m, 2H), 4.90 (bs, 2H), 4.69 (d, J = 6.1 Hz, 2H).
用與上文所述製備化合物13a之實例12(流程3)相同之程序製備實例13－16(化合物13b－13e)。Example 13-16 (Compound 13b-13e) was prepared by the same procedure as Example 12 (Scheme 3) for the preparation of compound 13a.
N－(2－胺基－苯基)－4－[(4－噻吩－3－基－嘧啶－2－基胺基)－甲基]－苯甲醯胺(16a)步驟1：2－氯－4－噻吩－3－基－嘧啶(14) 向3－噻吩酸(500 mg，3.91 mmol)及2,4－二氯嘧啶(1.16 g，7.81 mmol)之乙腈(20 mL)溶液中加入0.4 M Na2 CO3 (20 mL)溶液隨後加入Pd(PPh3 )4 (450 mg，0.39 mmol)。將該懸浮液脫氣且在氮氣下於90℃下加熱16小時，冷卻，濃縮且以EtOAc萃取之。相繼以NH4 Cl飽和溶液、鹽水洗滌有機層，將其經無水Na2 SO4 乾燥，過濾且加以濃縮。由急驟層析法經矽膠(EtOAc/CH2 Cl2 ：2/98)純化殘餘物產生標題化合物14(680 mg，3.46 mmol，產率88%)。1 H NMR：(400 MHz,CDCl3 )δ(ppm)：8.56(d,J＝5.2 Hz,1H),8.19(dd,J＝3.2,1.2 Hz,1H),7.66(dd,J＝5.2,1.2 Hz,1H),7.46(d,J＝5.2 Hz,1H),7.43(dd,J＝5.2,2.8 Hz,1H)。 N-(2-Amino-phenyl)-4-[(4-thien-3-yl-pyrimidin-2-ylamino)-methyl]-benzamide (16a) Step 1: 2-Chlorine 4-thiophen-3-yl-pyrimidine (14) to 3-thiophene Add 0.4 M Na 2 CO 3 (20 mL) solution to a solution of acid (500 mg, 3.91 mmol) and 2,4-dichloropyrimidine (1.16 g, 7.81 mmol) in acetonitrile (20 mL) followed by Pd (PPh 3 ) 4 (450 mg, 0.39 mmol). The suspension was degassed and heated at 90 <0>C for 16 h under EtOAc. Sequentially with saturated NH 4 Cl solution, the organic layer was washed with brine, which was dried over anhydrous Na 2 SO 4, filtered and concentrated. By flash chromatography over silica gel (EtOAc / CH 2 Cl 2: 2/98) to give the title compound the residue was purified 14 (680 mg, 3.46 mmol, 88% yield). 1 H NMR: (400 MHz, CDCl 3 ) δ (ppm): 8.56 (d, J = 5.2 Hz, 1H), 8.19 (dd, J = 3.2, 1.2 Hz, 1H), 7.66 (dd, J = 5.2, 1.2 Hz, 1H), 7.46 (d, J = 5.2 Hz, 1H), 7.43 (dd, J = 5.2, 2.8 Hz, 1H).
步驟2：4－[(4－噻吩－3－基－嘧啶－2－基胺基)－甲基]－苯甲酸甲酯(15) 向14(680 mg，3.46 mmol)及4－(胺甲基)苯甲酸甲酯(686 mg，4.51 mmol)之無水1,4－二噁烷(10 mL)溶液中加入DIPEA(1.50 mL，8.65 mmol)，且在130℃下於一密閉試管中加熱該混合物48小時。真空下移除溶劑且以EtOAc/Et2 O之混合物濕磨該殘餘物形成固體，其經過濾收集且加以乾燥。由急驟層析法經矽膠(EtOAc/CH2 Cl2 ：30/70)純化該材料產生標題化合物15(540 mg，1.66 mmol，產率48%)。1 H NMR：(400 MHz,DMSO－d6 )δ(ppm)：8.29－8.23(m,2H),7.88(d,J＝8.4 Hz,2H),7.84－7.77(m,1H),7.69－7.59(m,2H),7.52－7.43(m,2H),7.03(d,J＝5.2 Hz,1H),4.60(d,J＝6.4 Hz,2H),3.81(s,3H)。 Step 2: 4-[(4-Thien-3-yl-pyrimidin-2-ylamino)-methyl]-benzoic acid methyl ester (15) to 14 (680 mg, 3.46 mmol) and 4- (amine A DIPEA (1.50 mL, 8.65 mmol) was added to a solution of methyl benzoate (686 mg, 4.51 mmol) in anhydrous 1,4-dioxane (10 mL) and heated at 130 ° C in a closed tube The mixture was 48 hours. And the solvent removed in vacuo to EtOAc / Et 2 O mixture of the residue was triturated solid was formed which was collected by filtration and dried. By flash chromatography over silica gel (EtOAc / CH 2 Cl 2: 30/70) to give the title compound 15 the purified material (540 mg, 1.66 mmol, 48% yield). 1 H NMR: (400 MHz, DMSO-d 6 ) δ (ppm): 8.29-8.23 (m, 2H), 7.78 (d, J = 8.4 Hz, 2H), 7.84-7.77 (m, 1H), 7.69- 7.59 (m, 2H), 7.52-7.43 (m, 2H), 7.03 (d, J = 5.2 Hz, 1H), 4.60 (d, J = 6.4 Hz, 2H), 3.81 (s, 3H).
步驟3：N－(2－胺基－苯基)－4－[(4－噻吩－3－基－嘧啶－2－基胺基)－甲基]－苯甲醯胺(16a) 依照與實例2中相同之程序自15以步驟3及4(流程2)兩步獲得呈乳白色固體之標題化合物16a(實例17)。1 H NMR：(400 MHz,DMSO－d6)δ(ppm)：9.59(s,1H),8.32－8.27(m,2H),7.91(d,J＝8.0 Hz,2H),7.83(t,J＝6.4 Hz,1H),7.71(d,J＝4.8 Hz,1H),7.68－7.63(m,1H),7.54－7.44(m,2H),7.15(d,J＝8.0 Hz,1H),7.06(d,J＝5.2 Hz,1H),6.96(td,J＝7.6,1.6 Hz,1H),6.77(dd,J＝8.0,1.2 Hz,1H),6.59(td,J＝7.6,1.2 Hz,1H),4.89(s,2H),4.62(d,J＝6.4 Hz,2H)。 Step 3: N-(2-Amino-phenyl)-4-[(4-thiophen-3-yl-pyrimidin-2-ylamino)-methyl]-benzamide (16a) according to and examples The same procedure as in 2 gave the title compound 16a (Example 17) as a milky white solid from 15 steps in steps 3 and 4 (Scheme 2). 1 H NMR: (400 MHz, DMSO-d6) δ (ppm): 9.59 (s, 1H), 8.32 - 8.27 (m, 2H), 7.91 (d, J = 8.0 Hz, 2H), 7.83 (t, J = 6.4 Hz, 1H), 7.71 (d, J = 4.8 Hz, 1H), 7.68-7.63 (m, 1H), 7.54-7.44 (m, 2H), 7.15 (d, J = 8.0 Hz, 1H), 7.06 (d, J = 5.2 Hz, 1H), 6.96 (td, J = 7.6, 1.6 Hz, 1H), 6.77 (dd, J = 8.0, 1.2 Hz, 1H), 6.59 (td, J = 7.6, 1.2 Hz, 1H), 4.89 (s, 2H), 4.62 (d, J = 6.4 Hz, 2H).
用與上文所述製備化合物16a之實例17(流程4)相同之程序製備實例18－24(化合物16b－16h)。Examples 18-24 (Compounds 16b-16h) were prepared using the same procedure as Example 17 (Scheme 4) for the preparation of compound 16a.
N－(2－胺基－苯基)－4－({4－[4－(2－嗎啉－4－基－乙氧基)－苯基]－嘧啶－2－基胺基}－甲基)－苯甲醯胺(22a)步驟1：2－氯－4－(4－甲氧基－苯基)－嘧啶(17) 向4－甲氧基苯基酸(3.0 g，19.7 mmol)及2,4－二氯嘧啶(5.9 g，39.0 mmol)之無水乙腈(120 mL)溶液中加入0.4 MNa2 CO3 溶液(120 mL)隨後加入Pd(PPh3 )4 (400 mg，0.35 mmol)。將該懸浮液脫氣且於90℃下加熱16小時，使其冷卻且濃縮產生沉澱物，該沉澱物經過濾收集，以水洗滌，乾燥且由急驟層析法經矽膠(EtOAc/CH2 Cl2 ：5/95)純化產生標題化合物17(4.25 g，19.3 mmol，產率97%)。 N-(2-Amino-phenyl)-4-({4-[4-(2-morpholin-4-yl-ethoxy)-phenyl]-pyrimidin-2-ylamino}-- -benzamide (22a) Step 1: 2-Chloro-4-(4-methoxy-phenyl)-pyrimidine (17) to 4-methoxyphenyl To a solution of acid (3.0 g, 19.7 mmol) and 2,4-dichloropyrimidine (5.9 g, 39.0 mmol) in dry acetonitrile (120 mL) was added 0.4 M Na 2 CO 3 (120 mL) and then Pd (PPh 3 ) 4 (400 mg, 0.35 mmol). The suspension was degassed and heated at 90 ° C for 16 hours, allowed to cool and concentrated to give a precipitate which was collected by filtration, washed with water, dried and purified by flash chromatography (EtOAc/CH 2 Cl 2 :5/95) purification gave the title compound 17 (4.25 g, 19.3 mmol, yield 97%).
步驟2：4－(2－氯－嘧啶－4－基)－苯酚(18) 在0℃下向17(3.7 g，16.8 mmol)之無水二氯甲烷(42 mL)溶液中加入三溴化硼(3.17 mL，33.5 mmol)。在室溫下劇烈攪拌該混合物16小時，將其冷卻至0℃。向其中傾入冰水且繼續攪拌30分鐘。濃縮該反應混合物產生沉澱物，過濾收集沉澱物，將其以水洗滌，乾燥且由急驟層析法經矽膠(MeOH/CH2 Cl2 ：2/98)產生標題化合物18(3.28 g，15.9 mmol，產率94%)。1H NMR：(400 MHz,DMSO－d6 )δ(ppm)：10.26(s,1H),8.66(d,J＝5.6 Hz,1H),8.05(td,J＝8.4,1.6 Hz,2H),7.96(d,J＝5.6 Hz,1H),6.90(td,J＝8.4,1.6 Hz,2H)。 Step 2: 4-(2-Chloro-pyrimidin-4-yl)-phenol (18) Add boron tribromide to a solution of 17 (3.7 g, 16.8 mmol) anhydrous dichloromethane (42 mL) at 0 °C (3.17 mL, 33.5 mmol). The mixture was stirred vigorously at room temperature for 16 hours and cooled to 0 °C. Pour ice water into it and continue to stir for 30 minutes. The reaction mixture was concentrated to produce a precipitate, the precipitate was collected by filtration, washed with water, dried and purified by flash chromatography on silica gel (MeOH / CH 2 Cl 2: 2/98) to produce the title compound by 18 (3.28 g, 15.9 mmol , yield 94%). 1H NMR: (400 MHz, DMSO-d 6 ) δ (ppm): 10.26 (s, 1H), 8.66 (d, J = 5.6 Hz, 1H), 8.05 (td, J = 8.4, 1.6 Hz, 2H), 7.96 (d, J = 5.6 Hz, 1H), 6.90 (td, J = 8.4, 1.6 Hz, 2H).
步驟3：4－{2－[4－(2－氯－嘧啶－4－基)－苯氧基]－乙基}－嗎啉(19) 向18(1.8 g，8.71 mmol)之丙酮(80 mL)溶液中分別加入4－(2－氯乙基)嗎啉氫氯酸鹽(1.95 g，10.5 mmol)、碘化鉀(360 mg，2.2 mmol)及碳酸鉀(6.0 g，44.0 mmol)。將反應混合物回流16小時且加以濃縮。以水稀釋殘餘物且以EtOAc萃取含水相兩次。將合併之有機萃取物經無水Na2 SO4 乾燥，過濾及濃縮形成殘餘物，由急驟層析法經矽膠(EtOAc/CH2 Cl2 ，50/50至MeOH/CH2 Cl2 ：2/98)純化該殘餘物產生標題化合物19(2.7 g，8.4 mmol，產率96%)。 Step 3: 4-{2-[4-(2-Chloro-pyrimidin-4-yl)-phenoxy]-ethyl}-morpholine (19) to 18 (1.8 g, 8.71 mmol) of acetone (80) To the solution was added 4-(2-chloroethyl)morpholine hydrochloride (1.95 g, 10.5 mmol), potassium iodide (360 mg, 2.2 mmol) and potassium carbonate (6.0 g, 44.0 mmol). The reaction mixture was refluxed for 16 h and concentrated. The residue was diluted with water and the aqueous phase was extracted twice with EtOAc. The combined organic extracts were dried over anhydrous Na 2 SO 4, filtered and concentrated to form the residue by flash chromatography over silica gel (EtOAc / CH 2 Cl 2, 50/50 to MeOH / CH 2 Cl 2: 2/98 The residue was purified to give the title compound 19 (2.7 g, 8.4
步驟4：4－({4－[4－(2－嗎啉－4－基－乙氧基)－苯基]－嘧啶－2－基胺基}－甲基)－苯甲酸甲酯(20) 向19(2.7 g，8.4 mmol)及4－(胺甲基)苯甲酸甲酯氫氯酸鹽(2.7 g，13.5 mmol)之無水甲苯(33 mL)溶液中加入碳酸銫(8.2 g，25.3 mmol)隨後加入Pd2 (dba)3 (464 mg，0.51 mmol)及外消旋－BINAP(473 mg，0.76 mmol)。將溶液脫氣且在100℃下加熱16小時。將反應混合物在水與EtOAc之間分溶且分離該等相。將有機層依次以鹽水洗滌，經無水Na2 SO4 乾燥，過濾及濃縮形成殘餘物，由急驟層析法經矽膠(MeOH/CH2 Cl2 ：2/98)純化該殘餘物產生標題化合物20(1.9 g，4.2 mmol，產率50%)。 Step 4: 4-({4-[4-(2-morpholin-4-yl-ethoxy)-phenyl]-pyrimidin-2-ylamino}-methyl)-benzoic acid methyl ester (20 To a solution of 19 (2.7 g, 8.4 mmol) and methyl 4-(aminomethyl)benzoate hydrochloride (2.7 g, 13.5 mmol) in dry toluene (33 mL) Methyl) followed by the addition of Pd 2 (dba) 3 (464 mg, 0.51 mmol) and racemic-BINAP (473 mg, 0.76 mmol). The solution was degassed and heated at 100 ° C for 16 hours. The reaction mixture was partitioned between water and EtOAc and the phases were separated. The organic layer was successively washed with brine, dried over anhydrous Na 2 SO 4, filtered and concentrated to form a residue, (MeOH / CH 2 Cl 2 : 2/98) over silica gel by flash chromatography of the residue was purified to give the title compound 20 (1.9 g, 4.2 mmol, yield 50%).
步驟5：4－({4－[4－(2－嗎啉－4－基－乙氧基)－苯基]－嘧啶－2－基胺基}－甲基)－苯甲酸(21) 向20(1.9 g，4.2 mmol)溶於THF(8 mL)、MeOH(8 mL)及水(4 mL)之混合物中的溶液加入NaOH(373 mg，9.3 mmol)。將混合物在40℃下加熱16小時，接著藉由加入1 N HCl將其酸化至pH 6，濃縮，且在高度真空下乾燥產生不經進一步純化即使用之標題化合物21。 Step 5: 4-({4-[4-(2-morpholin-4-yl-ethoxy)-phenyl]-pyrimidin-2-ylamino}-methyl)-benzoic acid (21) 20 (1.9 g, 4.2 mmol) of a solution of THF (8 mL), MeOH (8 mL) and water (4 mL). The mixture was heated at 40 <0>C for 16 h then EtOAc (EtOAc) EtOAc.
步驟6：N－(2－胺基－苯基)－4－({4－[4－(2－嗎啉－4－基－乙氧基)－苯基]－嘧啶－2－基胺基}－甲基)－苯甲醯胺(22a) 向21(來自先前步驟之粗產物)之無水乙腈(50 mL)溶液中加入1,2－苯二胺(1.83 g，16.9 mmol)，隨後添加Et3 N(2.65 mL，19.0 mmol)、HOBt.H2 O(1.03 g，7.6 mmol)及EDCl.HCl(1.62 g，8.5 mmol)。室溫下攪拌該混合物72小時，將其過濾移除鹽且濃縮濾液形成殘餘物，其由急驟層析法經矽膠(MeOH/CH2 Cl2 ：2/98至5/95)純化。以EtOAc/CH2 Cl2 之混合物濕磨該材料產生呈白色固體之標題化合物22a(696 mg，1.3 mmol，經2個步驟之產率31%)。1 H NMR：(400 MHz,DMSO－d6 )δ(ppm)：9.55(s,1H),8.25(d,J＝5.2 Hz,1H),8.00(d,J＝8.4 Hz,2H),7.88(d,J＝8.4 Hz,2H),7.77(t,J＝6.4 Hz,1H),7.44(d,J＝7.2 Hz,2H),7.11(d,J＝7.6 Hz,1H),7.07(d,J＝5.2 Hz,1H),7.01(d,J＝8.4 Hz,2H),6.92(td,J＝7.6,1.6 Hz,1H),6.73(dd,J＝8.0,1.6 Hz,1H),6.55(td,J＝7.4,1.2 Hz,1H),4.85(s,2H),4.61(d,J＝5.6 Hz,2H),4.13(t,J＝5.6 Hz,2H),3.55(t,J＝4.4 Hz,4H),2.68(t,J＝5.6 Hz,2H),2.45(t,J＝4.4 Hz,4H)。 Step 6: N-(2-Amino-phenyl)-4-({4-[4-(2-morpholin-4-yl-ethoxy)-phenyl]-pyrimidin-2-ylamino) }-Methyl)-benzamide (22a) To a solution of 21 (crude from the previous step) in anhydrous acetonitrile (50 mL) was added 1,2-phenylenediamine (1.83 g, 16.9 mmol). Et 3 N (2.65 mL, 19.0 mmol), HOBt.H 2 O (1.03 g, 7.6 mmol) and EDCl.HCl (1.62 g, 8.5 mmol). The mixture was stirred at room temperature for 72 hours, it was filtered to remove salts and the filtrate was concentrated to form a residue which silica gel (MeOH / CH 2 Cl 2: 2/98 to 5/95) was purified by flash chromatography. In EtOAc / CH 2 Cl 2 mixture of the wet-milled to produce the title material as a white solid of compound 22a (696 mg, 1.3 mmol, yield over the two steps 31%). 1 H NMR: (400 MHz, DMSO-d 6 ) δ (ppm): 9.55 (s, 1H), 8.25 (d, J = 5.2 Hz, 1H), 8.00 (d, J = 8.4 Hz, 2H), 7.88 (d, J = 8.4 Hz, 2H), 7.77 (t, J = 6.4 Hz, 1H), 7.44 (d, J = 7.2 Hz, 2H), 7.11 (d, J = 7.6 Hz, 1H), 7.07 (d , J=5.2 Hz, 1H), 7.01 (d, J=8.4 Hz, 2H), 6.92 (td, J=7.6, 1.6 Hz, 1H), 6.73 (dd, J=8.0, 1.6 Hz, 1H), 6.55 (td, J = 7.4, 1.2 Hz, 1H), 4.85 (s, 2H), 4.61 (d, J = 5.6 Hz, 2H), 4.13 (t, J = 5.6 Hz, 2H), 3.55 (t, J = 4.4 Hz, 4H), 2.68 (t, J = 5.6 Hz, 2H), 2.45 (t, J = 4.4 Hz, 4H).
N－(2－胺基－苯基)－4－({4－[3－(2－二甲基胺基－乙氧基)－苯基]－嘧啶－2－基胺基}－甲基)－苯甲醯胺氫氯酸鹽(22b) 依照與實例25中步驟1－6(流程5)相同之程序自3－甲氧苯基酸開始且在步驟3中使用2－(二甲胺)乙基氯氫氯酸鹽作為烷基化試劑以6步獲得標題化合物22b。藉由將其溶解於MeOH與EtOAc之混合物中且添加1 N HCl之Et2 O溶液獲得該化合物之氫氯酸鹽。濾出白色沉澱物，將其以EtOAc洗滌且在高真空下乾燥。1H NMR(400 MHz,CDCl3 )δ(ppm)：8.31(d,J＝5.2 Hz,1H),7.91(d,J＝7.6 Hz,2H),7.50(d,J＝7.6 Hz,1H),7.45(d,J＝8.4 Hz,2H),7.32(t,J＝8.0 Hz,1H),7.34－7.28(m,1H),7.05(td,J＝7.6,1.6 Hz,1H),7.00－6.96(m,1H),6.97(d,J＝4.8 Hz,1H),6.82－6.77(m,2H),6.04(bs,1H),4.74(d,J＝6.0 Hz,2H),4.11(t,J＝5.2 Hz,2H),2.94(t,J＝5.2 Hz,2H),2.45(s,6H)。 N-(2-Amino-phenyl)-4-({4-[3-(2-dimethylamino-ethoxy)-phenyl]-pyrimidin-2-ylamino}-methyl - Benzylamine Hydrochloride (22b) from 3-methoxyphenyl according to the same procedure as Step 1-6 (Scheme 5) in Example 25. The acid was started and the title compound 22b was obtained in 6 steps using 2-(dimethylamine)ethyl chloride hydrochloride as the alkylating reagent. The hydrochloride salt of the compound was obtained by dissolving it in a mixture of MeOH and EtOAc and adding 1 N HCl in Et 2 O. The white precipitate was filtered, washed with EtOAc and dried EtOAc. 1H NMR (400 MHz, CDCl 3 ) δ (ppm): 8.31 (d, J = 5.2 Hz, 1H), 7.91 (d, J = 7.6 Hz, 2H), 7.50 (d, J = 7.6 Hz, 1H), 7.45 (d, J = 8.4 Hz, 2H), 7.32 (t, J = 8.0 Hz, 1H), 7.34 - 7.28 (m, 1H), 7.05 (td, J = 7.6, 1.6 Hz, 1H), 7.00-6.96 (m, 1H), 6.97 (d, J = 4.8 Hz, 1H), 6.82-6.77 (m, 2H), 6.04 (bs, 1H), 4.74 (d, J = 6.0 Hz, 2H), 4.11 (t, J = 5.2 Hz, 2H), 2.94 (t, J = 5.2 Hz, 2H), 2.45 (s, 6H).
N－(2－胺基－苯基)－4－({4－[3－(2－嗎啉－4－基－乙氧基)－苯基]－嘧啶－2－基胺基}－甲基)－苯甲醯胺(22c) 根據與實例25中(步驟1－6，流程5)相同之程序自3－甲氧苯基酸開始且在步驟3中使用4－(2－氯乙基)嗎啉氫氯酸鹽作為烷基化試劑以6步獲得標題化合物22c。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.54(s,1H),8.31(d,J＝5.2 Hz,1H),7.92－7.83(m,3H),7.65－7.52(m,2H),7.45(d,J＝6.4 Hz,2H),7.35(t,J＝8.4 Hz,1H),7.15(d,J＝5.2 Hz,1H),7.11(d,J＝6.8 Hz,1H),7.03(d,J＝6.8 Hz,1H),6.92(td,J＝7.6,1.2 Hz,1H),6.73(dd,J＝7.6,1.2 Hz,1H),6.55(t,J＝6.8 Hz,1H),4.85(s,2H),4.61(d,J＝6.0 Hz,2H),4.1－44.06(m,2H),3.55(t,J＝4.8 Hz,4H),2.70(t,J＝5.6 Hz,2H),2.50－2.44(m,4H)。 N-(2-Amino-phenyl)-4-({4-[3-(2-morpholin-4-yl-ethoxy)-phenyl]-pyrimidin-2-ylamino}-- Benzo-benzamide (22c) according to the same procedure as in Example 25 (Step 1-6, Scheme 5) from 3-methoxyphenyl The acid was started and the title compound 22c was obtained in 6 steps using 4-(2-chloroethyl)morpholine hydrochloride as the alkylating reagent. 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.54 (s, 1H), 8.31 (d, J = 5.2 Hz, 1H), 7.92-7.83 (m, 3H), 7.65-7.52 (m) , 2H), 7.45 (d, J = 6.4 Hz, 2H), 7.35 (t, J = 8.4 Hz, 1H), 7.15 (d, J = 5.2 Hz, 1H), 7.11 (d, J = 6.8 Hz, 1H) ), 7.03 (d, J = 6.8 Hz, 1H), 6.92 (td, J = 7.6, 1.2 Hz, 1H), 6.73 (dd, J = 7.6, 1.2 Hz, 1H), 6.55 (t, J = 6.8 Hz) , 1H), 4.85 (s, 2H), 4.61 (d, J = 6.0 Hz, 2H), 4.1-44.06 (m, 2H), 3.55 (t, J = 4.8 Hz, 4H), 2.70 (t, J = 5.6 Hz, 2H), 2.50-2.44 (m, 4H).
N－(2－胺基－苯基)－4－({4－[4－(2－二甲基胺基－乙氧基)－苯基]－嘧啶－2－基胺基}－甲基)－苯甲醯胺(22d) 根據與實例25中(步驟1－6，流程5)相同之程序自4－甲氧苯基酸開始且在步驟3中使用2－(二甲基胺)乙基氯氫氯酸鹽作為烷基化試劑以6步獲得標題化合物22d。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：10.52－10.35(bs,1H),9.97(s,1H),8.29(d,J＝5.2 Hz,1H),8.06(d,J＝8.8 Hz,2H),7.96(d,J＝8.0 Hz,2H),8.10－7.85(m,1H),7.52－7.40(m,2H),7.31(t,J＝7.6 Hz,1H),7.20－7.00(m,5H),6.96－6.88(m,1H),4.70－4.58(m,2H),4.45－4.38(t,J＝4.8 Hz,2H),3.54－3.46(m,2H),2.83(s,6H)。 N-(2-Amino-phenyl)-4-({4-[4-(2-dimethylamino-ethoxy)-phenyl]-pyrimidin-2-ylamino}-methyl -benzimidamide (22d) according to the same procedure as in Example 25 (Step 1-6, Scheme 5) from 4-methoxyphenyl Starting with the acid and using 2-(dimethylamine)ethyl chloride hydrochloride as the alkylating reagent in step 3, the title compound 22d was obtained in 6 steps. 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 10.52-10.35 (bs, 1H), 9.97 (s, 1H), 8.29 (d, J = 5.2 Hz, 1H), 8.06 (d, J) = 8.8 Hz, 2H), 7.96 (d, J = 8.0 Hz, 2H), 8.10-7.85 (m, 1H), 7.52-7.40 (m, 2H), 7.31 (t, J = 7.6 Hz, 1H), 7.20 -7.00 (m, 5H), 6.96-6.88 (m, 1H), 4.70-4.58 (m, 2H), 4.45-4.38 (t, J = 4.8 Hz, 2H), 3.54-3.46 (m, 2H), 2.83 (s, 6H).
N－(2－胺基－苯基)－4－[(4－吡嗪－2－基－嘧啶－2－基胺基)－甲基]－苯甲醯胺(26a)步驟1：3－二甲胺基－1－吡嗪－2－基－丙烯酮(23a) 在110℃下加熱N,N－二甲基甲醯胺二甲縮醛(10.9 mL，81.8 mmol)及Et3 N(5.7 mL)中之乙醯吡嗪(5 g，40.9 mmol)溶液16小時。停止加熱且使其冷卻至室溫時形成沉澱物。以第三丁基甲基醚稀釋該懸浮液；過濾分離且以第三丁基甲基醚洗滌該固體。以相同溶劑濕磨該材料，將其濾出且乾燥以產生呈黃色固體之標題化合物23a(5.9 g，33.3 mmol，產率81%)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.09(s,1H),8.73(d,J＝2.4 Hz,1H),8.66(dd,J＝2.4,1.6 Hz,1H),7.84(d,J＝12.4 Hz,1H),6.30－6.20(m,1H),3.19(s,3H),2.93(s,3H)。 N-(2-Amino-phenyl)-4-[(4-pyrazin-2-yl-pyrimidin-2-ylamino)-methyl]-benzamide (26a) Step 1: 3- Dimethylamino-1-pyrazin-2-yl-propenone (23a) was heated at 110 ° C with N,N-dimethylformamide dimethyl acetal (10.9 mL, 81.8 mmol) and Et 3 N ( A solution of acetaminopyrazine (5 g, 40.9 mmol) in 5.7 mL) for 16 hours. The heating was stopped and allowed to cool to room temperature to form a precipitate. The suspension was diluted with tert-butyl methyl ether; it was isolated by filtration and washed with tri-butyl methyl ether. The material was triturated with EtOAc (EtOAc)EtOAc. 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.09 (s, 1H), 8.73 (d, J = 2.4 Hz, 1H), 8.66 (dd, J = 2.4, 1.6 Hz, 1H), 7.84 (d, J = 12.4 Hz, 1H), 6.30-6.20 (m, 1H), 3.19 (s, 3H), 2.93 (s, 3H).
步驟2：二水合4－胍甲基－苯甲酸甲基酯(24) 向4－(胺甲基)苯甲酸甲酯氫氯酸鹽(5 g)之無水乙醇(25 mL)溶液中加入1H－吡唑－1－羧脒氫氯酸鹽(4.4 g)，隨後添加DIPEA(13.0 mL)，且將混合物回流3小時。真空下移除乙醇。在劇烈攪拌下向剩餘黏性油緩緩加入NaHCO3 飽和溶液(50 mL)，隨後添加300 mL水(所得pH為9)。形成白色固體且繼續攪拌1小時。濾出該材料，將其以水(200 mL)及第三丁基甲基醚(50 mL)洗滌，且經乾燥產生白色粉末狀標題化合物24(4.7 g，91%)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：7.91(d,J＝8.0 Hz,2H),7.39(d,J＝8.0 Hz,2H),4.39(s,2H),3.83(s,3H),3.80－2.80[m,4H＋2H2 O(由元素分析測定)]。 Step 2: 4-Hydroxymethyl-benzoic acid methyl ester (24) was added to a solution of 4-(aminomethyl)benzoic acid methyl ester hydrochloride (5 g) in absolute ethanol (25 mL). Pyrazole-1-carboxyindole hydrochloride (4.4 g) followed by DIPEA (13.0 mL) and the mixture was refluxed for 3 h. The ethanol was removed under vacuum. A saturated NaHCO 3 solution (50 mL) was slowly added to the remaining viscous oil with vigorous stirring, followed by the addition of 300 mL of water (the resulting pH was 9). A white solid formed and stirring was continued for 1 hour. The material was filtered, washed with EtOAc EtOAc m. 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 7.91 (d, J = 8.0 Hz, 2H), 7.39 (d, J = 8.0 Hz, 2H), 4.39 (s, 2H), 3.83 ( s, 3H), 3.80-2.80 [m, 4H + 2H 2 O (determined by elemental analysis)].
步驟3：方法A：4－[(4－吡嗪－2－基－嘧啶－2－基胺基)－甲基]－苯甲酸(25a) 攪拌24(6.2 g，25.4 mmol)及23a(3.0 g，16.9 mmol)之無水甲醇(40 mL)懸浮液且將其加熱至回流歷時10分鐘，接著緩緩添加甲氧基鈉95%(3.65 g，67.6 mmol)之甲醇(40 mL)溶液。回流24小時後，添加20 mL溶於水中之2.5 N NaOH且再保持回流24小時。使混合物冷卻至室溫，減壓移除甲醇，添加100 mL水且以AcOEt萃取所得混合物。分離含水相且以2 N HCl酸化至pH 5－6以形成沉澱物，該沉澱物經過濾收集、以水沖洗且經乾燥產生呈白色固體之所要羧酸25a (4.55 g，14.8 mmol，87%)。 Step 3: Method A: 4-[(4-Pyrazin-2-yl-pyrimidin-2-ylamino)-methyl]-benzoic acid (25a) stirred 24 (6.2 g, 25.4 mmol) and 23a (3.0 A suspension of g, 16.9 mmol) in dry methanol (40 mL) was then warmed to reflux for 10 min then a solution of sodium methoxide 95% (3.65 g, 67.6 mmol) in methanol (40 mL). After refluxing for 24 hours, 20 mL of 2.5 N NaOH dissolved in water was added and maintained at reflux for a further 24 hours. The mixture was allowed to cool to room temperature, methanol was removed under reduced pressure, 100 mL water was added and the mixture was extracted with AcOEt. The aqueous phase was separated and acidified to pH 5-6 with 2N EtOAc to afford a precipitate which was collected by filtration, washed with water and dried to give the desired carboxylic acid 25a (4.55 g, 14.8 mmol, 87% ).
步驟4：N－(2－胺基－苯基)－4－[(4－吡嗪－2－基－嘧啶－2－基胺基)－甲基]－苯甲醯胺(26a) 向25a(1 g，3.3 mmol)之無水乙腈(35 mL)溶液中加入1,2－苯二胺(0.88 g，8.1 mmol)，隨後加入Et3 N(2.7 mL，19.1 mmol)、HOBt.H2 O(803 mg,5.9 mmol)及EDCl.HCl(1.89 g，9.9 mmol)。於室溫下攪拌該混合物16小時以形成懸浮液，其經過濾收集，相繼以MeCN、水及再次以MeCN洗滌，以MeOH濕磨，將其過濾及乾燥以產生標題化合物26a(730 mg，1.84 mmol，產率55%)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.57(s,1H),9.43(s,1H),8.75(d,J＝4.8 Hz,2H),8.49(d,J＝5.2 Hz,1H),8.11(t,J＝5.2 Hz,1H),7.91(d,J＝8.4 Hz,2H),7.59－7.37(m,2H),7.44(d,J＝5.2 Hz,1H),7.12(d,J＝7.6 Hz,1H),6.94(td,J＝7.6,1.6 Hz,1H),6.75(dd,J＝8.0,1.2 Hz,1H),6.57(t,J＝7.2 Hz,1H),4.86(s,2H),4.66(d,J＝5.2 Hz,2H)。 Step 4: N-(2-Amino-phenyl)-4-[(4-pyrazin-2-yl-pyrimidin-2-ylamino)-methyl]-benzamide (26a) to 25a (1 g, 3.3 mmol) of anhydrous acetonitrile (35 mL) was charged with 1,2-phenylenediamine (0.88 g, 8.1 mmol), followed by Et 3 N (2.7 mL, 19.1 mmol), HOBt.H 2 O (803 mg, 5.9 mmol) and EDCl.HCl (1.89 g, 9.9 mmol). The mixture was stirred at room temperature for 16 hours to give a suspension which was filtered, washed with EtOAc EtOAc EtOAc EtOAc EtOAc EtOAc M, yield 55%). 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.57 (s, 1H), 9.43 (s, 1H), 8.75 (d, J = 4.8 Hz, 2H), 8.49 (d, J = 5.2 Hz, 1H), 8.11 (t, J = 5.2 Hz, 1H), 7.91 (d, J = 8.4 Hz, 2H), 7.59-7.37 (m, 2H), 7.44 (d, J = 5.2 Hz, 1H), 7.12 (d, J = 7.6 Hz, 1H), 6.94 (td, J = 7.6, 1.6 Hz, 1H), 6.75 (dd, J = 8.0, 1.2 Hz, 1H), 6.57 (t, J = 7.2 Hz, 1H) ), 4.86 (s, 2H), 4.66 (d, J = 5.2 Hz, 2H).
N－(2－胺基－苯基)－4－[(4－吡啶－2－基－嘧啶－2－基胺基)－甲基]－苯甲醯胺(26b) 依照與實例29步驟1、2及4(流程6)相同之程序製備化合物26b。步驟3中使用方法B：步驟3，方法B：4－[(4－吡啶－2－基－嘧啶－2－基胺基)－甲基]－苯甲酸(25b) 向24(0.85 g,4.83 mmol)及3－二甲胺－1－吡啶－2－基－丙烯酮23b(1.0 g，4.83 mmol)之異丙二醇(20 mL)溶液中加入分子篩(0.2 g，4，粉末，>5μm)。將反應混合物回流16小時接著經矽藻土墊過濾該渾濁溶液。將母液濃縮至其一半體積形成固體，過濾收集該固體且加以乾燥產生淺黃色結晶材料(0.62 g，1.94 mmol，產率40%)。將該化合物(0.456 g，1.43 mmol)溶解於THF(3 mL)、MeOH(3 mL)及水(1.5 mL)之混合物中，接著添加NaOH(0.125 g，3.14 mmol)且在40℃下攪拌該反應混合物16小時，將其冷卻至室溫，藉由加入1NHCl(3.2 mL)將其酸化至pH 5－6，且濃縮移除有機溶劑。形成沉澱物，過濾收集該沉澱物，將其水洗滌及乾燥產生標題化合物25b(0.542 g，1.37 mmol，產率96%)。 N-(2-Amino-phenyl)-4-[(4-pyridin-2-yl-pyrimidin-2-ylamino)-methyl]-benzamide (26b) according to Step 29 of Example 29. Compounds 26b were prepared in the same procedure as 2, 4 (Scheme 6). Step B is used in Step 3 : Step 3, Method B: 4-[(4-Pyridin-2-yl-pyrimidin-2-ylamino)-methyl]-benzoic acid (25b) to 24 (0.85 g, 4.83 Molecular sieve (0.2 g, 4) in a solution of mmol) and 3-dimethylamine-1-pyridin-2-yl-propenone 23b (1.0 g, 4.83 mmol) in isopropyl diol (20 mL) , powder, >5 μm). The reaction mixture was refluxed for 16 hours then the turbid solution was filtered thru a pad. The mother liquor was concentrated to half its volume to form a solid which was collected by filtration and dried to give a pale-yellow crystalline material (0.62 g, 1.94 mmol, yield 40%). This compound (0.456 g, 1.43 mmol) was dissolved in THF (3 mL), MeOH (3 mL) The reaction mixture was allowed to cool to room temperature and then acidified to pH 5-6 by adding 1N HCl (3.2 mL) and concentrated to remove organic solvent. A precipitate formed which was collected by filtration, washed with water and dried to give the title compound 25b (0.542 g, 1.37 mmol, yield 96%).
使用與化合物26a所述相同之程序(實例29，流程6)製備實例31－33(化合物26b－26d)。Examples 31-33 (compounds 26b-26d) were prepared using the same procedure as described for compound 26a (Example 29, Scheme 6).
5－{2－[4－(2－胺基－苯胺甲醯基)－苄胺基]－嘧啶－4－基}－吡啶－2－甲酸二甲醯胺(29a)步驟1：6－二甲基胺甲醯基－煙酸(27) 在室溫下於氮氣下經10分鐘向吡啶－2,5－二甲酸二甲酯(10.1 g，51.6 mmol)及MgBr2 (4.75 g，25.8 mmol)之THF(200 mL)懸浮液中逐滴加入二甲胺(51.6 mL，103.2 mmol，在THF中2 N)溶液。將反應混合物攪拌隔夜且以1 N HCl(52 mL)及H2 O(50 mL)中止，以EtOAc(200 mL x 3)萃取之。以鹽水洗滌有機相，將其經Na2 SO4 乾燥，過濾及濃縮。將粗殘餘物溶解於DMF(10 mL)、AcOEt(50 mL)、MeOH(20 mL)及DCM(50 mL)之混合物中，部分蒸發形成之溶液以產生結晶材料，其經過濾移除。收集母液且蒸發形成固體，該固體溶於THF(67 mL)及MeOH(67 mL)之混合物中。向該溶液加入溶於H2 O(33.5 mL)中之NaOH(2.95 g，73.7 mmol)。將反應物在40℃加熱幾小時，酸化(pH 3)形成固體沉澱物，該沉澱經過濾收集及乾燥產生標題化合物27(4.937 g，經兩個步驟之產率50%)。 5-{2-[4-(2-Amino-anilinemethanyl)-benzylamino]-pyrimidin-4-yl}-pyridine-2-carboxylic acid dimethyl hydrazine (29a) Step 1: 6- Methylamine-mercapto-nicotinic acid (27) to pyridine-2,5-dicarboxylic acid dimethyl ester (10.1 g, 51.6 mmol) and MgBr 2 (4.75 g, 25.8 mmol ) at room temperature under nitrogen for 10 min. A solution of dimethylamine (51.6 mL, 103.2 mmol, 2 N in THF) was added dropwise to a THF (200 mL) suspension. The reaction mixture was stirred overnight and with 1 N HCl (52 mL) and H 2 O (50 mL) quenched with EtOAc (200 mL x 3) The extracted. The organic phase was washed with brine, the dried over Na 2 SO 4, filtered and concentrated. The crude residue was dissolved in a mixture of DMF (10 mL), EtOAc (50 mL), MeOH (20 mL) and DCM (50 mL) and partially evaporated to give a crystalline material which was removed by filtration. The mother liquor was collected and evaporated to a solid which was dissolved in a mixture of THF (67 mL) and MeOH (67 mL). To this solution was added dissolved in H 2 O (33.5 mL) in of NaOH (2.95 g, 73.7 mmol) . The reaction was heated at 40 <0>C for several hours and acidified (pH 3) to give a solid precipitate which was collected by filtration and dried to give the title compound 27 (4.937 g, 50% yield from two steps).
步驟2：5－乙醯基－吡啶－2－甲酸二甲醯胺(28) 將27(4.937 g，25.41 mmol)、SOCl2 (2.41 mL，33 mmol)及DMF(0.9 mL，5.1 mmol)之CH2 Cl2 (51 mL)懸浮液回流3小時。將反應混合物冷卻至室溫且蒸乾。令殘餘物懸浮於甲苯中且添加Et3 N(4.25 mL，30.5 mmol)。將所得懸浮液經2小時以套管***至經攪拌且預形成之丙二酸二甲酯(3.49 mL，30.5 mmol)、MgCl2 (1.742 g，18.3 mmol)、Et3 N(10.2 mL，73.2 mmol)之甲苯(25 mL)懸浮液。將所得反應混合物攪拌隔夜且以1 N HCl(50 mL)及水(50 mL)中止並以EtOAc(200 mL x 3)萃取之。將合併之有機層以鹽水(100 mL)洗滌，經Na2 SO4 乾燥及過濾。蒸乾濾液產生褐色油狀物(5.21 g)，將該油狀物溶解於DMSO(15.4 mL)及H2 O(0.62 mL)且接著在130℃下加熱2小時，冷卻至室溫，將其以H2 O(100 mL)稀釋，以EtOAc萃取。以鹽水(100 mL)洗滌該有機層，使其經Na2 SO4 乾燥，過濾且加以濃縮。由急驟層析法經矽膠(AcOEt/己烷，70/30至100/0)純化該殘餘物產生呈褐色結晶固體之標題化合物28(430 mg，產率9%)。 Step 2: 5-Ethyl-pyridine-2-carboxylic acid dimethylamine (28) 27 (4.937 g, 25.41 mmol), SOCl 2 (2.41 mL, 33 mmol) and DMF (0.9 mL, 5.1 mmol) The CH 2 Cl 2 (51 mL) suspension was refluxed for 3 h. The reaction mixture was cooled to room temperature and evaporated to dryness. So the residue was suspended in toluene and add Et 3 N (4.25 mL, 30.5 mmol). The resulting suspension was cannulated into the stirred and preformed dimethyl malonate (3.49 mL, 30.5 mmol), MgCl 2 (1.742 g, 18.3 mmol), Et 3 N (10.2 mL, 73.2) over 2 hours. A suspension of toluene (25 mL) in mmol). The reaction mixture was stirred with EtOAc EtOAc (EtOAc) The combined organic layers were (100 mL) and washed with brine, dried over Na 2 SO 4 and filtered. The filtrate was evaporated to dryness to give a brown oil (5.21 g), which was dissolved in DMSO (15.4 mL) and H 2 O (0.62 mL) and then heated at 130 ° C for 2 hours, cooled to room temperature, diluted with H 2 O (100 mL), and extracted with EtOAc. The organic layer was washed with brine (100 mL) was washed, dried over Na 2 SO 4, filtered and concentrated. The residue was purified by flash chromatography eluting elut elut elut elut elut elut
步驟3：5－{2－[4－(2－胺基－苯胺甲醯基)－苄胺基]－嘧啶－4－基}－吡啶－2－甲酸二甲醯胺(29a) 依照與實例29中步驟1、3(方法B)及4(流程6)相同之程序自28以4個步驟獲得呈乳白色固體之標題化合物29a(實例34)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.57(s,1H),9.20(s,1H),8.50(m,1H),8.41(d,J＝4.9 Hz,1H),8.04(m,1H),7.89(d,J＝7.8 Hz,2H),7.65(m,1H),7.46(m,2H),7.29(d,J＝5.1 Hz,1H),7.11(d,J＝7.4 Hz,1H),6.93(m,1H),6.74(m,1H),6.56(m,1H),4.86(s,2H),4.65(m,2H),3.02(s,3H),2.94(s,3H)。 Step 3: 5-{2-[4-(2-Amino-anilinemethanyl)-benzylamino]-pyrimidin-4-yl}-pyridine-2-carboxylic acid dimethylamine (29a) according to and examples The same procedure as Steps 1 and 3 (Method B) and 4 (Scheme 6) in 29 gave the title compound 29a (Example 34) as a milky white solid from 28. 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.57 (s, 1H), 9.20 (s, 1H), 8.50 (m, 1H), 8.41 (d, J = 4.9 Hz, 1H), 8.04 (m, 1H), 7.89 (d, J = 7.8 Hz, 2H), 7.65 (m, 1H), 7.46 (m, 2H), 7.29 (d, J = 5.1 Hz, 1H), 7.11 (d, J) = 7.4 Hz, 1H), 6.93 (m, 1H), 6.74 (m, 1H), 6.56 (m, 1H), 4.86 (s, 2H), 4.65 (m, 2H), 3.02 (s, 3H), 2.94 (s, 3H).
N－(2－胺基－苯基)－4－({4－[6－(嗎啉－4－羰基)－吡啶－3－基]－嘧啶－2－基胺基}－甲基)－苯甲醯胺(29b) 使用與對化合物29a所述實例34(流程7)相同之程序製備標題化合物。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.55(s,1H),9.21(s,1H),8.52(m,1H),8.42(d,J＝4.9 Hz,1H),8.03(t,J＝6.2 Hz,1H),7.90(d,J＝8.0 Hz,2H),7.71(d,J＝8.0 Hz,1H),7.47(m,2H),7.29(d,J＝5.1 Hz,1H),7.12(d,J＝7.6 Hz,1H),6.93(m,1H),6.74(d,J＝7.8 Hz,1H),6.56(m,1H),4.85(s,2H),4.64(d,J＝4.9 Hz,2H),3.66(s,4H),3.55(m,2H),3.44(m,2H)。 N-(2-Amino-phenyl)-4-({4-[6-(morpholin-4-carbonyl)-pyridin-3-yl]-pyrimidin-2-ylamino}-methyl)- The title compound was prepared using the same procedure as described in Example 34 (Scheme 7) for compound 29a. 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.55 (s, 1H), 9.21. (s, 1H), 8.52 (m, 1H), 8.42 (d, J = 4.9 Hz, 1H), 8.03 (t, J = 6.2 Hz, 1H), 7.90 (d, J = 8.0 Hz, 2H), 7.71 (d, J = 8.0 Hz, 1H), 7.47 (m, 2H), 7.29 (d, J = 5.1) Hz, 1H), 7.12 (d, J = 7.6 Hz, 1H), 6.93 (m, 1H), 6.74 (d, J = 7.8 Hz, 1H), 6.56 (m, 1H), 4.85 (s, 2H), 4.64 (d, J = 4.9 Hz, 2H), 3.66 (s, 4H), 3.55 (m, 2H), 3.44 (m, 2H).
N－(2－胺基－苯基)－4－{[4－(2－甲基－吡啶－3－基)－嘧啶－2－基胺基]－甲基}－苯甲醯胺(31a)步驟1：1－(2－甲基－吡啶－3－基)－乙酮(30a) 依照與實例34(步驟2流程7)相同之程序自2－甲基煙酸以92%產率獲得呈褐色結晶固體之標題化合物30a。 N-(2-Amino-phenyl)-4-{[4-(2-methyl-pyridin-3-yl)-pyrimidin-2-ylamino]-methyl}-benzamide (31a The step 1: 1-(2-methyl-pyridin-3-yl)-ethanone (30a) was obtained from 2-methylnicotinic acid in 92% yield according to the same procedure as in Example 34 (Step 2 of Step 2). The title compound 30a was obtained as a brown crystalline solid.
步驟2：N－(2－胺基－苯基)4－{[4－(2－甲基－吡啶－3－基)－嘧啶－2－基胺基]－甲基}－苯甲醯胺(31a) 依照與實例29步驟1、3(方法B)及4(流程6)相同之程序自30a以4步獲得呈乳白色固體之標題化合物31a(實例36)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.57(s,1H),8.46(m,1H),8.40(m,1H),7.54(t,J＝6.4 Hz,1H),7.88(d,J＝8.2 Hz,2H),7.75(d,J＝6.3 Hz,1H),7.40(d,J＝7.8 Hz,2H),7.29(m,1H),7.11(d,J＝7.2 Hz,1H),6.93(m,1H),6.79(d,J＝4.9 Hz,1H),6.73(dd,J＝8.0,1.4 Hz,1H),6.55(m,1H),4.86(s,2H),4.48(d,J＝6.3 Hz,2H),2.66－2.82(m,3H)。 Step 2: N-(2-Amino-phenyl)4-{[4-(2-methyl-pyridin-3-yl)-pyrimidin-2-ylamino]-methyl}-benzamide (31a) The title compound 31a (yield 36) was obtained as a creamy white solid from <RTI ID=0.0>> 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.57 (s, 1H), 8.46 (m, 1H), 8.40 (m, 1H), 7.54 (t, J = 6.4 Hz, 1H), 7.88 (d, J = 8.2 Hz, 2H), 7.75 (d, J = 6.3 Hz, 1H), 7.40 (d, J = 7.8 Hz, 2H), 7.29 (m, 1H), 7.11 (d, J = 7.2) Hz, 1H), 6.93 (m, 1H), 6.79 (d, J = 4.9 Hz, 1H), 6.73 (dd, J = 8.0, 1.4 Hz, 1H), 6.55 (m, 1H), 4.86 (s, 2H) ), 4.48 (d, J = 6.3 Hz, 2H), 2.66-2.82 (m, 3H).
N－(2－胺基－苯基)－4－[(4－吡啶－4－基－嘧啶－2－基胺基)－甲基]－苯甲醯胺(31b) 使用與用於化合物31a之描述(實例36，流程8)相同之程序製備標題化合物31b。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.56(s,1H),8.70(d,J＝5.3 Hz,2H),8.44(d,J＝5.1 Hz,1H),8.05(t,J＝6.3 Hz,1H),7.98(d,J＝6.1 Hz,2H),7.90(d,J＝8.2 Hz,2H),7.47(bs,2H),7.27(d,J＝5.1 Hz,1H),7.12(d,J＝7.6 Hz,1H),6.93(m,1H),6.74(dd,J＝8.0,1.4 Hz,1H),6.56(m,1H),4.86(s,2H),4.64(d,J＝5.9 Hz,2H)。 N-(2-Amino-phenyl)-4-[(4-pyridin-4-yl-pyrimidin-2-ylamino)-methyl]-benzamide (31b) was used with compound 31a Description (Example 36, Scheme 8) The same procedure was used to prepare the title compound 31b. 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.56 (s, 1H), 8.70 (d, J = 5.3 Hz, 2H), 8.44 (d, J = 5.1 Hz, 1H), 8.05 ( t, J = 6.3 Hz, 1H), 7.98 (d, J = 6.1 Hz, 2H), 7.90 (d, J = 8.2 Hz, 2H), 7.47 (bs, 2H), 7.27 (d, J = 5.1 Hz, 1H), 7.12 (d, J = 7.6 Hz, 1H), 6.93 (m, 1H), 6.74 (dd, J = 8.0, 1.4 Hz, 1H), 6.56 (m, 1H), 4.86 (s, 2H), 4.64 (d, J = 5.9 Hz, 2H).
N－(2－胺基－苯基)－4－{[4－(6－嗎啉－4－基－吡啶－3－基)－嘧啶－2－基胺基]－甲基}－苯甲醯胺(34a)步驟1：1－(6－氯－吡啶－3－基)－乙酮(32) 在室溫下於氮氣下攪拌丙二酸二甲酯(7.8 mL，68.3 mmol)、MgCl2 (3.872 g，40.7 mmol)、Et3 N(19.1 mL，136.9 mmol)之甲苯(15 mL)懸浮液2小時。經由套管向該混合物中加入氯化6－氯－煙鹼醯基(3.872 g，40.7 mmol)及Et3 N(4.25 mL，30.5 mmol)之甲苯(46 mL)懸浮液。將所得反應混合物攪拌隔夜，以1 N HCl(100 mL)及水(100 mL)中止且以EtOAc萃取之。以鹽水(100 mL)洗滌有機層，使其經Na2 SO4 乾燥，過濾及濃縮產生白色結晶材料(6.5 g)，將該材料溶解於DMSO(9 mL)與H2 O(0.37 mL)之混合物中，在130℃下加熱5小時，冷卻至室溫且以水(10 mL)處理之。形成沉澱物，其經過濾收集、以水沖洗及乾燥產生呈淺黃色結晶固體之標題化合物32(1.8 g，產率28%)。 N-(2-Amino-phenyl)-4-{[4-(6-morpholin-4-yl-pyridin-3-yl)-pyrimidin-2-ylamino]-methyl}-benzene Indoleamine (34a) Step 1: 1 - (6-Chloro-pyridin-3-yl)-ethanone (32) Stir dimethyl malonate (7.8 mL, 68.3 mmol), MgCl at room temperature under nitrogen. 2 (3.872 g, 40.7 mmol) , Et 3 N (19.1 mL, 136.9 mmol) of toluene (15 mL) suspension for 2 hours. To this mixture was added via cannula chloride 6- chloro - nicotine acyl (3.872 g, 40.7 mmol) and Et 3 N (4.25 mL, 30.5 mmol) of toluene (46 mL) suspension. The resulting reaction mixture was stirred with EtOAc EtOAc m. The organic layer was washed with brine (100 mL), it was dried over Na 2 SO 4, filtered and concentrated to give a white crystalline material (6.5 g), the material was dissolved in DMSO (9 mL) and H 2 O (0.37 mL) of The mixture was heated at 130 ° C for 5 hours, cooled to room temperature and treated with water (10 mL). Precipitate was formed which was collected by filtration, washed with water and dried to afford title compound 32 (1.8 g, yield 28%).
步驟2：1－(6－嗎啉－4－基－吡啶－3－基)－乙酮(33a) 使32(1.25 g，7.9 mmol)及嗎啉(2.20 mL，25.2 mmol)之EtOH(22 mL)溶液回流12小時且接著蒸乾。將該殘餘物溶解於EtOAc(200 mL)中，以飽和NaHCO3 (50 mL x 2)及鹽水(50 mL)洗滌、經Na2 SO4 乾燥、過濾及蒸發產生呈淺黃色固體之標題化合物33a(1.66 g，定量產率)。 Step 2: 1-(6-morpholin-4-yl-pyridin-3-yl)-ethanone (33a) gave 32 (1.25 g, 7.9 mmol) and morpholine (2.20 mL, 25.2 mmol) of EtOH (22 The mL) solution was refluxed for 12 hours and then evaporated to dryness. The residue was dissolved in EtOAc (200 mL), washed with saturated NaHCO 3 (50 mL x 2) and brine (50 mL), dried over Na 2 SO 4, filtered, and evaporated to give the title compound 33a as a pale yellow solid of (1.66 g, quantitative yield).
步驟3：N－(2－胺基－苯基)－4－{[4－(6－嗎啉－4－基－吡啶－3－基)－嘧啶－2－基胺基]－甲基}－苯甲醯胺(34a) 依照與實例29步驟1、3(方法B)及4(流程6)相同之程序自33a以4步獲得呈乳白色固體之標題化合物34a(實例38)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.57(s,1H),8.84(s,1H),8.24(d,J＝5.3 Hz,1H),8.18(m,1H),7.90(d,J＝7.3 Hz,2H),7.78(m,1H),7.46(d,J＝7.8 Hz,2H),7.13(m,1H),7.08(d,J＝5.3 Hz,1H),6.95(d,J＝7.6 Hz,1H),6.91(m,1H),6.75(d,J＝7.8 Hz,1H),5.57(dd,J＝7.2,7.6 Hz,1H),4.87(s,2H),4.61(d,J＝6.1 Hz,2H),3.70(m,4H),3.56(m,4H)。 Step 3: N-(2-Amino-phenyl)-4-{[4-(6-morpholin-4-yl-pyridin-3-yl)-pyrimidin-2-ylamino]-methyl} - Benzalamine (34a) The title compound 34a (Example 38) was obtained as a creamy solid from EtOAc (m. 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.57 (s, 1H), 8.84 (s, 1H), 8.24 (d, J = 5.3 Hz, 1H), 8.18 (m, 1H), 7.90 (d, J = 7.3 Hz, 2H), 7.78 (m, 1H), 7.46 (d, J = 7.8 Hz, 2H), 7.13 (m, 1H), 7.08 (d, J = 5.3 Hz, 1H), 6.95 (d, J = 7.6 Hz, 1H), 6.91 (m, 1H), 6.75 (d, J = 7.8 Hz, 1H), 5.57 (dd, J = 7.2, 7.6 Hz, 1H), 4.87 (s, 2H) ), 4.61 (d, J = 6.1 Hz, 2H), 3.70 (m, 4H), 3.56 (m, 4H).
N－(2－胺基－苯基)－4－({4－[6－(4－甲基－六氫吡嗪－1－基)－吡啶－3－基]－嘧啶－2－基胺基}－甲基)－苯甲醯胺(34b) 使用與對化合物34a所述(實例38，流程9)相同之程序製備標題化合物34b。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.55(s,1H),8.81(d,J＝2.2 Hz,1H),8.22(d,J＝5.3 Hz,1H),8.14(t,J＝8.6 Hz,1H),7.89(d,J＝8.2 Hz,2H),7.75(t,J＝6.2 Hz,1H),7.45(d,J＝8.0 Hz,2H),7.12(d,J＝7.4 Hz,1H),7.05(d,J＝5.3 Hz,1H),6.93(m,1H),6.88(d,J＝8.2 Hz,1H),6.74(dd,J＝8.0,1.4 Hz,1H),6.56(m,1H),4.87(s,2H),4.62(d,J＝6.5 Hz,2H),3.59(m,4H),2.38(m,4H),2.21(s,3H)。 N-(2-Amino-phenyl)-4-({4-[6-(4-methyl-hexahydropyrazin-1-yl)-pyridin-3-yl]-pyrimidin-2-ylamine The title compound 34b was prepared using the same procedure as described for compound 34a (Example 38, Scheme 9). 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.55 (s, 1H), 8.81 (d, J = 2.2 Hz, 1H), 8.22 (d, J = 5.3 Hz, 1H), 8.14 ( t, J = 8.6 Hz, 1H), 7.89 (d, J = 8.2 Hz, 2H), 7.75 (t, J = 6.2 Hz, 1H), 7.45 (d, J = 8.0 Hz, 2H), 7.12 (d, J = 7.4 Hz, 1H), 7.05 (d, J = 5.3 Hz, 1H), 6.93 (m, 1H), 6.88 (d, J = 8.2 Hz, 1H), 6.74 (dd, J = 8.0, 1.4 Hz, 1H), 6.56 (m, 1H), 4.87 (s, 2H), 4.62 (d, J = 6.5 Hz, 2H), 3.59 (m, 4H), 2.38 (m, 4H), 2.21. (s, 3H).
N－(2－胺基－苯基)－4－{[4－(1－氧基－吡啶－3－基)－嘧啶－2－基胺基]－甲基}－苯甲醯胺(36)步驟1：1－(1－氧基－吡啶－3－基)－乙酮(35) 在室溫下向1－吡啶－3－基－乙酮(1.00 g,8.3 mmol)之CH2 Cl2 (8.3 mL)溶液中加入MTO(甲基三氧化錸，113 mg，0.45 mmol)且將反應混合物冷卻至15℃。經20分鐘逐滴添加H2 O2 (30%，1.13 mL，9.96 mmol)水溶液且在15℃下攪拌反應混合物5小時並將其冷卻至0℃。添加Na2 S2 O3 (20%，20 mL)水溶液，攪拌該混合物10分鐘接著以EtOAc萃取之。收集含水層且將其冷凍－乾燥形成固體材料，由急驟層析法經矽膠(MeOH/CH2 Cl2 ，10/90)純化該固體材料產生標題化合物35(1.3 g，定量產量)。 N-(2-Amino-phenyl)-4-{[4-(1-oxy-pyridin-3-yl)-pyrimidin-2-ylamino]-methyl}-benzamide (36) Step 1: 1-(1-oxy-pyridin-3-yl)-ethanone (35) to 1-pyridin-3-yl-ethanone (1.00 g, 8.3 mmol) of CH 2 Cl at rt M (Methyl Antimony Trioxide, 113 mg, 0.45 mmol) was added to a solution of 2 (8.3 mL) and the mixture was cooled to 15 °C. An aqueous solution of H 2 O 2 (30%, 1.13 mL, 9.96 mmol) was added dropwise over 20 min and the mixture was stirred at 15 ° C for 5 hr and cooled to 0 ° C. Aqueous Na 2 S 2 O 3 (20%, 20 mL) was added and the mixture was stirred for 10 min then extracted with EtOAc. The aqueous layer was collected and the freeze - dried to form a solid material produced by flash silica gel chromatography (MeOH / CH 2 Cl 2, 10/90) the solid material was purified title compound 35 (1.3 g, quantitative yield).
步驟2：N－(2－胺基－苯基)4－([4－(1－氧基－吡啶－3－基)－嘧啶－2－基胺基]－甲基}－苯甲醯胺(36) 根據與實例29步驟1、3(方法B)及4(流程6)相同之程序自35以4步獲得呈淺黃色固體之標題化合物36(實例40)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.61(s,1H),8.81(bs,1H),8.45(m,1H),8.33(d,J＝5.1 Hz,1H),8.12(bs,1H),7.99(d,J＝7.8 Hz,1H),7.93(d,J＝8.0 Hz,2H),7.55(t,J＝7.0 Hz,1H),7.50(m,2H),7.29(d,J＝5.1 Hz,1H),7.16(d,J＝7.4 Hz,1H),6.97(m,1H),6.78(dd,J＝8.0,1.2 Hz,1H),6.60(m,1H),4.91(s,2H),4.67(d,J＝6.3 Hz,2H)。 Step 2: N-(2-Amino-phenyl)4-([4-(1-oxy-pyridin-3-yl)-pyrimidin-2-ylamino]-methyl}-benzamide (36) The title compound 36 (Example 40) was obtained as a pale yellow solid from <RTI ID=0.0>#</RTI></RTI> DMSO-d 6 ) δ (ppm): 9.61 (s, 1H), 8.81 (bs, 1H), 8.45 (m, 1H), 8.33 (d, J = 5.1 Hz, 1H), 8.12 (bs, 1H), 7.99 (d, J = 7.8 Hz, 1H), 7.93 (d, J = 8.0 Hz, 2H), 7.55 (t, J = 7.0 Hz, 1H), 7.50 (m, 2H), 7.29 (d, J = 5.1) Hz, 1H), 7.16 (d, J = 7.4 Hz, 1H), 6.97 (m, 1H), 6.78 (dd, J = 8.0, 1.2 Hz, 1H), 6.60 (m, 1H), 4.91 (s, 2H) ), 4.67 (d, J = 6.3 Hz, 2H).
N－(2－胺基－苯基)－4－({4－[4－(2－二甲胺基－乙氧基)－3－氟－苯基]－嘧啶－2－基胺基}－甲基)－苯甲醯胺(41)步驟1：4－(4－苄氧基－3－氟－苯基)－2－氯－嘧啶(37) 依照與實例25步驟1(流程5)相同之程序自4－苄氧基－3－氟苯酸開始獲得標題化合物37。1 H NMR(400 MHz,CDCl3 )δ(ppm)：8.60(d,J＝5.2 Hz,1H),7.90(dd,J＝12.0,2.4 Hz,1H),7.84－7.80(m,1H),7.54(d,J＝5.2 Hz,1H),7.48－7.32(m,5H),7.10(t,J＝8.4 Hz,1H),5.24(s,2H)。 N-(2-Amino-phenyl)-4-({4-[4-(2-dimethylamino-ethoxy)-3-fluoro-phenyl]-pyrimidin-2-ylamino} -Methyl)-benzamide (41) Step 1: 4-(4-Benzyloxy-3-fluoro-phenyl)-2-chloro-pyrimidine (37) according to Example 25, Step 1 (Scheme 5) The same procedure from 4-benzyloxy-3-fluorobenzene The acid began to obtain the title compound 37. 1 H NMR (400 MHz, CDCl 3 ) δ (ppm): 8.60 (d, J = 5.2 Hz, 1H), 7.90 (dd, J = 12.0, 2.4 Hz, 1H), 7.84-7.80 (m, 1H), 7.54 (d, J = 5.2 Hz, 1H), 7.48-7.32 (m, 5H), 7.10 (t, J = 8.4 Hz, 1H), 5.24 (s, 2H).
步驟2：4－{[4－(4－苄氧基－3－氟－苯基)－嘧啶－2－基胺基]－甲基}－苯甲酸甲酯(38) 依照與實例17(步驟2，流程4)相同之程序自37獲得標題化合物38。1 H NMR(400 MHz,CDCl3 )δ(ppm)：8.23(d,J＝4.4 Hz,1H),7.98(d,J＝8.4 Hz,2H),7.78(dd,J＝12.4,2.0 Hz,1H),7.67(ddd,J＝8.0,2.0,1.2 Hz,1H),7.47－7.29(m,7H),7.01(t,J＝8.4 Hz,1H),6.88(d,J＝5.6 Hz,1H),6.06－5.95(m,1H),5.18(s,2H),4.75(d,J＝6.4 Hz,2H),3.90(s,3H)。 Step 2: 4-{[4-(4-Benzyloxy-3-fluoro-phenyl)-pyrimidin-2-ylamino]-methyl}-benzoic acid methyl ester (38) according to Example 17 (step 2. Scheme 4) The same procedure was obtained from 37 to give the title compound 38. 1 H NMR (400 MHz, CDCl 3 ) δ (ppm): 8.23 (d, J = 4.4 Hz, 1H), 7.78 (d, J = 8.4 Hz, 2H), 7.78 (dd, J = 12.4, 2.0 Hz, 1H), 7.67 (ddd, J=8.0, 2.0, 1.2 Hz, 1H), 7.47-7.29 (m, 7H), 7.01 (t, J = 8.4 Hz, 1H), 6.88 (d, J = 5.6 Hz, 1H) ), 6.06-5.95 (m, 1H), 5.18 (s, 2H), 4.75 (d, J = 6.4 Hz, 2H), 3.90 (s, 3H).
步驟3：4－{[4－(3－氟－4－羥基－苯基)－嘧啶－2－基胺基]－甲基}－苯甲酸甲酯(39) 在室溫下於氮氣下向38(950 mg，2.14 mmol)之EtOAc(100 mL)脫氣溶液中加入10% Pd/C(220 mg，0.21 mmol)。氫化該混合物5天(1 atm，氣球(balloon))，使其經矽藻土墊過濾，以MeOH及EtOAc沖洗，濃縮該濾液產生標題化合物39(450 mg，1.27 mmol，產率59%)。 Step 3: 4-{[4-(3-Fluoro-4-hydroxy-phenyl)-pyrimidin-2-ylamino]-methyl}-benzoic acid methyl ester (39) at room temperature under nitrogen To a degassed solution of 38 (950 mg, 2.14 mmol) in EtOAc (100 mL), 10% Pd / C (220 mg, 0.21 mmol). The mixture was hydrogenated for 5 days (1 atm, EtOAc) (EtOAc) elute
步驟4：4－({4－[4－(2－二甲胺－乙氧基)－3－氟－苯基]－嘧啶－2－基胺基}－甲基)－苯甲酸甲酯(40) 向39(450 mg，1.27 mmol)之丙酮(25 mL)溶液中加入2－(二甲基胺)乙基氯氫氯酸鹽(220 mg，1.53 mmol)，隨後添加碘化鉀(53 mg，0.32 mmol)及碳酸鉀(878 mg，6.35 mmol)。將反應混合物回流20小時，接著添加NH4 Cl飽和溶液，將混合物之pH調整至8且減壓移除丙酮。所形成之固體經過濾收集，以水洗滌，將其乾燥且由急驟層析法經矽膠(MeOH/DCM/NH4 OH：10/89/1)純化產生標題化合物40 (525 mg，1.24 mmol，產率97%)。 Step 4: 4-({4-[4-(2-Dimethylamine-ethoxy)-3-fluoro-phenyl]-pyrimidin-2-ylamino}-methyl)-benzoic acid methyl ester ( 40) To a solution of 39 (450 mg, 1.27 mmol) in acetone (25 mL) was added 2-(dimethylamine)ethylchlorohydrochloride (220 mg, 1.53 mmol), followed by potassium iodide (53 mg, 0.32 mmol) and potassium carbonate (878 mg, 6.35 mmol). The reaction mixture was refluxed for 20 hours, then a saturated solution of NH 4 Cl was added, the mixture was adjusted to pH 8 and acetone was removed under reduced pressure. The solid formed was collected by filtration, washed with water, dried and silica gel (MeOH / DCM / NH 4 OH : 10/89/1) by a flash chromatography purification to give the title compound 40 (525 mg, 1.24 mmol, Yield 97%).
步驟5：N－(2－胺基－苯基)－4－({4－[4－(2－二甲胺基－乙氧基)－3－氟－苯基]－嘧啶－2－基胺基}－甲基)－苯甲醯胺(41) 依照與實例25步驟5及6(流程5)相同之程序自40 獲得呈乳白色固體之標題化合物41(實例41)。1 H NMR：(400 MHz,DMSO－d6 )δ(ppm)：9.55(s,1H),8.27(d,J＝5.2 Hz,1H),7.95－7.75(m,5H),7.50－7.35(m,2H),7.25(t,J＝8.0 Hz,1H),7.14－7.06(m,2H),6.92(t,J＝7.6 Hz,1H),6.72(d,J＝8.0 Hz,1H),6.54(t,J＝7.6 Hz,1H),4.85(s,2H),4.60(d,J＝5.2 Hz,2H),4.17(t,J＝5.6 Hz,2H),2.66(t,J＝5.6 Hz,2H),2.21(s,6H)。 Step 5: N-(2-Amino-phenyl)-4-({4-[4-(2-dimethylamino-ethoxy)-3-fluoro-phenyl]-pyrimidin-2-yl) Amino}-methyl)-benzamide (41) The title compound 41 (Example 41) was obtained as a white solid from 40 from EtOAc. 1 H NMR: (400 MHz, DMSO-d 6 ) δ (ppm): 9.55 (s, 1H), 8.27 (d, J = 5.2 Hz, 1H), 7.95-7.75 (m, 5H), 7.50-7.35 ( m, 2H), 7.25 (t, J = 8.0 Hz, 1H), 7.14 - 7.06 (m, 2H), 6.92 (t, J = 7.6 Hz, 1H), 6.72 (d, J = 8.0 Hz, 1H), 6.54 (t, J = 7.6 Hz, 1H), 4.85 (s, 2H), 4.60 (d, J = 5.2 Hz, 2H), 4.17 (t, J = 5.6 Hz, 2H), 2.66 (t, J = 5.6 Hz, 2H), 2.21 (s, 6H).
N－(2－胺基－苯基)－4－{[4－(4－嗎啉－4－基甲基－苯基)－嘧啶－2－基胺基]－甲基}－苯甲醯胺(44)步驟1：4－(2－氯－嘧啶－4－基)－苯甲醛(42) 依照與實例25步驟1(流程5)相同之程序自4－甲醯基苯基酸開始獲得標題化合物42。1 H NMR：(400 MHz,CDCl3 )δ(ppm)：10.11(s,1H),8.73(d,J＝4.8 Hz,1H),8.27(td,J＝8.4,1.6 Hz,2H),8.03(td,J＝8.4,1.6 Hz,2H),7.73(d,J＝4.8 Hz,1H)。 N-(2-Amino-phenyl)-4-{[4-(4-morpholin-4-ylmethyl-phenyl)-pyrimidin-2-ylamino]-methyl}-benzamide Amine (44) Step 1: 4-(2-Chloro-pyrimidin-4-yl)-benzaldehyde (42) according to the same procedure as Example 25 Step 1 (Scheme 5) from 4-Methylphenyl The acid begins to give the title compound 42. 1 H NMR: (400 MHz, CDCl 3 ) δ (ppm): 10.11 (s, 1H), 8.73 (d, J = 4.8 Hz, 1H), 8.27 (td, J = 8.4, 1.6 Hz, 2H), 8.03 (td, J = 8.4, 1.6 Hz, 2H), 7.73 (d, J = 4.8 Hz, 1H).
步驟2：4－[4－(2－氯－嘧啶－4－基)－苄基]－嗎啉(43) 在室溫下向42(950 mg，4.35 mmol)及嗎啉(455 μL，5.21 mmol)之無水1,2－二氯乙烷(l0 mL)溶液中加入AcOH(2滴)，隨後添加NaBH(OAc)3 (1.1 g，5.21 mmol)且攪拌該混合物16小時。向反應混合物加入10%K2 CO3 溶液隨後添加二氯甲烷且分離該等相。以CH2 Cl2 －萃取含水相且使有機層經無水Na2 SO4 乾燥、過濾以形成殘餘物，由急驟層析法經矽膠(EtOAc/CH2 Cl2 ：20/80)純化該殘餘物產生標題化合物43(460 mg，1.59 mmol，產率36%)。 Step 2: 4-[4-(2-Chloro-pyrimidin-4-yl)-benzyl]-morpholine (43) at room temperature to 42 (950 mg, 4.35 mmol) and morpholine (455 μL, 5.21. To a solution of anhydrous 1,2-dichloroethane (10 mL) was added AcOH (2 drops), then NaBH(OAc) 3 (1.1 g, 5.21 mmol) was added and the mixture was stirred for 16 hours. A 10% K 2 CO 3 solution was added to the reaction mixture followed by the addition of dichloromethane and the phases were separated. In CH 2 Cl 2 - aqueous phase was extracted and the organic layer was dried over anhydrous Na 2 SO 4, filtered to form the residue by flash chromatography over silica gel (EtOAc / CH 2 Cl 2: 20/80) which was purified The title compound 43 (460 mg, 1.59 mmol, yield 36%) was obtained.
步驟3：N－(2－胺基－苯基)4－{[4－(4－嗎啉－4－基甲基－苯基)－嘧啶－2－基胺基]－甲基}－苯甲醯胺(44) 依照與實例2步驟2－4(流程2)相同之程序，自43以三步驟獲得標題化合物44，且藉由添加1 N HCl之Et2 O溶液將其溶解於二氯甲烷及EtOAc之混合物中而分離出該標題化合物之氫氯酸鹽。濾除沉澱物，將其以EtOAc洗滌且在高真空下乾燥。1 H NMR(400 MHz,CDCl3 )δ(ppm)：8.33(d,J＝5.2 Hz,1H),8.00－7.93(m,1H),7.95(d,J＝8.0 Hz,2H),7.85(d,J＝8.0 Hz,2H),7.47(d,J＝8.0 Hz,2H),7.44(d,J＝8.0 Hz,2H),7.29(d，J＝8.4 Hz,1H),7.07(td，J＝7.6,1.2 Hz,1H),7.00(d,J＝5.2 Hz,1H),6.85－6.78(m,2H),5.83(t,J＝6.0 Hz,1H),4.78(d,J＝6.0 Hz,2H),3.74(t,J＝4.0 Hz,4H),3.59(s,2H),2.51(bs,4H)。 Step 3: N-(2-Amino-phenyl)4-{[4-(4-morpholin-4-ylmethyl-phenyl)-pyrimidin-2-ylamino]-methyl}-benzene Methionine (44) The title compound 44 was obtained in the three steps from 43 in the same procedure as in Example 2 Steps 2-4 (Scheme 2), and dissolved in dichlorobenzene by adding 1 N HCl in Et 2 O. The hydrochloride salt of the title compound was isolated from a mixture of methane and EtOAc. The precipitate was filtered off, washed with EtOAc and dried under high vacuum. 1 H NMR (400 MHz, CDCl 3 ) δ (ppm): 8.33 (d, J = 5.2 Hz, 1H), 8.00 - 7.93 (m, 1H), 7.95 (d, J = 8.0 Hz, 2H), 7.85 ( d, J = 8.0 Hz, 2H), 7.47 (d, J = 8.0 Hz, 2H), 7.44 (d, J = 8.0 Hz, 2H), 7.29 (d, J = 8.4 Hz, 1H), 7.07 (td, J = 7.6, 1.2 Hz, 1H), 7.00 (d, J = 5.2 Hz, 1H), 6.85-6.78 (m, 2H), 5.83 (t, J = 6.0 Hz, 1H), 4.78 (d, J = 6.0) Hz, 2H), 3.74 (t, J = 4.0 Hz, 4H), 3.59 (s, 2H), 2.51 (bs, 4H).
N－(2－胺基－苯基)－4－[(2－吡啶－3－基－嘧啶－4－基胺基)－甲基]－苯甲醯胺(47)步驟1：4－[(2－氯－嘧啶－4－基胺基)－甲基]－苯甲酸甲酯(45a) 及4－[(4－氯－嘧啶－2－基胺基)－甲基]－苯甲酸甲酯(45b)將在異丙二醇(60 mL)及DMF(40 mL)中之2,4－二氯嘧啶(4.51 g，30.3 mmol)、4－胺甲基－苯甲酸甲酯(5.00 g，30.3 mmol)、DIPEA(10.4 mL，60.6 mmol)之混合物加熱回流5小時。蒸乾該反應混合物後由急驟層析法經矽膠(EtOAc/己烷：40/60→60/40＋1% Et3 N)純化該殘餘物產生標題化合物45a(3.454 g，產率41%)及45b(1.52 g，產率14%，受起始物質污染)。 N-(2-Amino-phenyl)-4-[(2-pyridin-3-yl-pyrimidin-4-ylamino)-methyl]-benzamide (47) Step 1: 4-[ (2-Chloro-pyrimidin-4-ylamino)-methyl]-benzoic acid methyl ester (45a) and 4-[(4-chloro-pyrimidin-2-ylamino)-methyl]-benzoic acid Ester (45b) 2,4-dichloropyrimidine (4.51 g, 30.3 mmol), 4-aminomethyl-benzoic acid methyl ester (5.00 g, 30.3) in isopropyl diol (60 mL) and DMF (40 mL) A mixture of mmol) and DIPEA (10.4 mL, 60.6 mmol) was heated to reflux for 5 h. The reaction mixture was evaporated to dryness by flash chromatography over silica gel: Purification of the residue (EtOAc / hexane 40/60 → 60/40 + 1% Et 3 N) to give the title compound 45a (3.454 g, 41% yield) and 45b (1.52 g, yield 14%, contaminated with starting material).
45a，1 H NMR(400 MHz,DMS0－d6 )δ(ppm)：8.45(bs,1H),7.90－7.94(m,3H),7.42(d,J＝8.4 Hz,2H),6.53(d,J＝5.9 Hz,1H),4.58(d,J＝5.3 Hz,2H),3.83(s,3H)。45a, 1 H NMR (400 MHz, DMS0-d 6 ) δ (ppm): 8.45 (bs, 1H), 7.90-7.94 (m, 3H), 7.42 (d, J = 8.4 Hz, 2H), 6.53 (d) , J = 5.9 Hz, 1H), 4.58 (d, J = 5.3 Hz, 2H), 3.83 (s, 3H).
45b，1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：8.27(t,J＝6.5 Hz,1H),8.21(bs,1H),7.88(d,J＝8.2 Hz,2H),7.40(d,J＝8.2 Hz,2H),6.69(d,J＝5.1 Hz，1H),4.55(bs,2H),3.82(s,3H)。 45b, 1 H NMR (400 MHz , DMSO-d 6) δ (ppm): 8.27 (t, J = 6.5 Hz, 1H), 8.21 (bs, 1H), 7.88 (d, J = 8.2 Hz, 2H), 7.40 (d, J = 8.2 Hz, 2H), 6.69 (d, J = 5.1 Hz, 1H), 4.55 (bs, 2H), 3.82 (s, 3H).
步驟2：4－[(2－吡啶－3－基－嘧啶－4－基胺基)－甲基]－苯甲酸甲酯(46) 在室溫下向3－吡啶環硼氧烷(189 mg，0.60 mmol)、45a(500 mg,1.81 mmol)、Pd(OAc)2 (41 mg,0.18 mmol)及PPh3 (95 mg，0.36 mmol)之DME(1.8 mL)懸浮液中加入Na2 CO3 溶液(590 mg溶解於最小量水中，5.60 mmol)。以氮氣吹掃該反應混合物且使其回流4天，蒸乾且由急驟層析法經矽膠(EtOAc/己烷：30/70＋1%Et3 N)純化產生呈淺黃色固體之標題化合物46(152 mg，產率26%)。 Step 2: 4-[(2-Pyridin-3-yl-pyrimidin-4-ylamino)-methyl]-benzoic acid methyl ester (46) to 3-pyridineboroxine at room temperature (189 mg , a solution of 0.60 mmol), 45a (500 mg, 1.81 mmol), Pd(OAc) 2 (41 mg, 0.18 mmol) and PPh 3 (95 mg, 0.36 mmol) in DME (1.8 mL) was added Na 2 CO 3 Solution (590 mg dissolved in a minimum amount of water, 5.60 mmol). The reaction mixture was purged with nitrogen and allowed to reflux for 4 days, evaporated to dryness and purified by flash chromatography on silica gel (EtOAc / hexane: 30/70 + 1% Et 3 N) as a pale yellow solid was purified title compound 46 (152 Mg, yield 26%).
步驟3：N－(2－胺基－苯基)－4－[(2－吡啶－3－基－嘧啶－4－基胺基)－甲基]－苯甲醯胺(47) 依照與實例34步驟3(反應3及4)(流程7)相同之程序自46以兩步獲得呈乳白色固體之標題化合物47(實例43)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.50(s,1H),9.30(d,J＝1.6 Hz,1H),8.54(dd,J＝4.7,1.6 Hz,1H),8.46(ddd，J＝8.0,2.0,2.0 Hz，1H),8.11(dd,J 6.3,6.1 Hz,2H),7.85(d,J＝8.2 Hz,2H),7.38－7.43(m,3H),7.05(m,1H),6.86(m,1H),6.67(dd,J＝7.9,1.5 Hz,1H),6.49(m,2H),4.79(s,2H),4.66(s,2H)。 Step 3: N-(2-Amino-phenyl)-4-[(2-pyridin-3-yl-pyrimidin-4-ylamino)-methyl]-benzamide (47) according to and examples 34 Step 3 (Reactions 3 and 4) (Scheme 7) The same procedure was obtained from the title compound 47 (Example 43) as a white solid. 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.50 (s, 1H), 9.30 (d, J = 1.6 Hz, 1H), 8.54 (dd, J = 4.7, 1.6 Hz, 1H), 8.46 (ddd, J=8.0, 2.0, 2.0 Hz, 1H), 8.11 (dd, J 6.3, 6.1 Hz, 2H), 7.85 (d, J = 8.2 Hz, 2H), 7.38-7.43 (m, 3H), 7.05 (m, 1H), 6.86 (m, 1H), 6.67 (dd, J = 7.9, 1.5 Hz, 1H), 6.49 (m, 2H), 4.79 (s, 2H), 4.66 (s, 2H).
N－(2－胺基－苯基)－4－[(2－嗎啉－4－基－嘧啶－4－基胺基)－甲基]－苯甲醯胺(49)步驟2：4－[(2－嗎啉－4－基－嘧啶－4－基胺基)－甲基]－苯甲酸甲酯(48a) 將一密閉燒瓶中異丙二醇(18.0 mL)中之45a(2.50 g，9.0 mmol)、嗎啉(0.95 mL，10.8 mmol)及i－Pr2 NEt(3.12 mmol)混合物在120℃下加熱隔夜且冷卻至室溫。所形成之沉澱經過濾收集，以異丙二醇沖洗及乾燥產生標題化合物48(2.96 g，定量產量)。 N-(2-Amino-phenyl)-4-[(2-morpholin-4-yl-pyrimidin-4-ylamino)-methyl]-benzamide (49) Step 2: 4- [(2-Morpholin-4-yl-pyrimidin-4-ylamino)-methyl]-benzoic acid methyl ester (48a) 45a (2.50 g, 9.0 in isopropyl diol (18.0 mL) in a closed flask The mixture of mmol), morpholine (0.95 mL, 10.8 mmol) and i-Pr 2 NEt (3.12 mmol) was heated overnight at 120 ° C and cooled to room temperature. The precipitate formed was collected by filtration, washed with EtOAc and dried to afford titled compound 48 (2.96 g, quantitative yield).
步驟3：N－(2－胺基－苯基)4－[(2－嗎啉－4－基－嘧啶－4－基胺基)－甲基]－苯甲醯胺(49) 依照與實例34步驟3(反應3及4)(流程7)相同之程序自48獲得呈乳白色固體之標題化合物49(實例44)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.56(s,1H),7.89(d,J＝8.2 Hz,2H),7.72(d,J＝5.7 Hz,1H),7.56(bs,1H),7.40(d,J＝8.2 Hz,2H),7.13(d,J＝7.8 Hz,1H),6.94(m,1H),6.75(m,1H),6.57(m,1H),5.83(bs,1H),4.87(s,2H),4.51(bs,2H),3.56(s,8H)。 Step 3: N-(2-Amino-phenyl)4-[(2-morpholin-4-yl-pyrimidin-4-ylamino)-methyl]-benzamide (49) according to and examples 34 Step 3 (Reactions 3 and 4) (Scheme 7) The same procedure was obtained from the title compound 49 (Example 44) as a white solid. 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.56 (s, 1H), 7.89 (d, J = 8.2 Hz, 2H), 7.72 (d, J = 5.7 Hz, 1H), 7.56 ( Bs, 1H), 7.40 (d, J = 8.2 Hz, 2H), 7.13 (d, J = 7.8 Hz, 1H), 6.94 (m, 1H), 6.75 (m, 1H), 6.57 (m, 1H), 5.83 (bs, 1H), 4.87 (s, 2H), 4.51 (bs, 2H), 3.56 (s, 8H).
N－(2－胺基－苯基)－4－{[4－(6－甲基硫基－吡啶－3－基)－嘧啶－2－基胺基]－甲基}－苯甲醯胺(53)步驟1：5－溴－2－甲硫基－吡啶(50) 在－78℃下於氮氣下向2,5－二溴吡啶(5.00 g，21.11 mmol)之無水甲苯(300 mL)經攪拌溶液中緩緩加入正丁基鋰溶液(10.13 mL，25.33 mmol，己烷中2.5 M)。在－78℃下2小時後，加入二硫甲基(2.47 mL，27.44 mmol)。在－78℃下攪拌該反應混合物1小時且使其溫至室溫，以飽和NH4 Cl中止形成一兩相系統。分離該有機層，將其以飽和NH4 Cl、H2 O及鹽水洗滌，經無水MgSO4 乾燥，過濾且濃縮之。由急驟層析法經矽膠(AcOEt/己烷，5/95)純化該殘餘物產生呈淺黃色含油液體之標題化合物50(2.74 g，13.43 mmol，產率64%)。1 H NMR(400 MHz,CDCl3 )δ(ppm)：8.49(dd,J＝2.3,0.6 Hz,1H),7.60(dd,J＝8.5,2.4 Hz,1H),7.09(dd,J＝8.6,0.8 Hz,1H),2.57(s,3H)。 N-(2-Amino-phenyl)-4-{[4-(6-methylthio-pyridin-3-yl)-pyrimidin-2-ylamino]-methyl}-benzamide (53) Step 1: 5-Bromo-2-methylthio-pyridine (50) to 2,5-dibromopyridine (5.00 g, 21.11 mmol) in anhydrous toluene (300 mL) at -78 ° C under nitrogen. A solution of n-butyllithium (10.13 mL, 25.33 mmol, 2.5 M in hexane) was slowly added to the stirred solution. After 2 hours at -78 °C, dithiomethyl (2.47 mL, 27.44 mmol) was added. For 1 h and the reaction mixture was allowed to warm to room temperature and stirred at -78 ℃, with saturated NH 4 Cl the suspension to form a two-phase system. The organic layer was separated, washed with saturated NH 4 Cl, H 2 O and brine, dried over anhydrous MgSO 4, filtered and concentrated. The residue was purified by flash chromatography eluting elut elut elut elut elut elut elut elut elut 1 H NMR (400 MHz, CDCl 3 ) δ (ppm): 8.49 (dd, J = 2.3, 0.6 Hz, 1H), 7.60 (dd, J = 8.5, 2.4 Hz, 1H), 7.09 (dd, J = 8.6 , 0.8 Hz, 1H), 2.57 (s, 3H).
步驟2：5－(2－甲硫基－吡啶基)－ 酸(51) 在－40℃下氮氣下向溶於無水甲苯/THF(20 mL/5 mL)混合物中之50(2.74 g，13.43 mmol)及硼酸三異丙酯(3.72 mL，16.11 mmol)經攪拌溶液中逐滴加入正丁基鋰(6.98 mL，17.45 mmol，己烷中2.5 M)溶液。在－40℃下攪拌1小時後，將混合物溫至室溫且以2 N HCl中止。過濾所得懸浮液；以H2 O及AcOEt沖洗該沉澱物。以1 N NaOH中和(pH 7)濾液且以AcOEt加以萃取。合併有機層及沉澱物，蒸發溶劑且以MeCN－MeOH濕磨固體殘餘物產生呈淺黃色固體之標題化合物51(1.84 g，10.88 mmol，產率81%)。 Step 2: 5-(2-Methylthio-pyridyl)- The acid (51) was subjected to 50 (2.74 g, 13.43 mmol) and triisopropyl borate (3.72 mL, 16.11 mmol) in a mixture of anhydrous toluene/THF (20 mL / 5 mL ) at -40 ° C under nitrogen. A solution of n-butyllithium (6.98 mL, 17.45 mmol, 2.5 M in hexane) was added dropwise to the stirred solution. After stirring at -40 °C for 1 hour, the mixture was warmed to room temperature and quenched with 2 N HCl. The resulting suspension was filtered; AcOEt and H 2 O to rinse the precipitate. The filtrate was neutralized (pH 7) with 1 N NaOH and extracted with AcOEt. The organic layer was combined with EtOAc EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj
步驟3：4－{[4－(6－甲硫基－吡啶－3－基)－嘧啶－2－基胺基]－甲基}－苯甲酸(52) 在室溫下向51(593 mg，3.51 mmol)、45b(500 mg，1.80 mmol)及Na2 CO3 溶液(15 mL，0.4 M)之混合物的乙腈(15 mL)經攪拌脫氣懸浮液中加入Pd(PPh3 )4 (126 mg，0.11 mmol)。在氮氣下將反應混合物在90－95℃下加熱24小時。接著加入1 M NaOH(量)且繼續再加熱2小時。冷卻至室溫後將反應混合物過濾，以AcOEt萃取濾液，將含水層收集、過濾、濃縮且以2 N HCl(pH為5－6)酸化。形成沉澱物，其經過濾收集及乾燥以產生呈米色固體之標題化合物52(396 mg，1.12 mmol，產率62%)。 Step 3: 4-{[4-(6-Methylthio-pyridin-3-yl)-pyrimidin-2-ylamino]-methyl}-benzoic acid (52) at room temperature to 51 (593 mg , a mixture of 3.51 mmol), 45b (500 mg, 1.80 mmol) and a solution of Na 2 CO 3 (15 mL, 0.4 M) in acetonitrile (15 mL) was added to a stirred degassed suspension and Pd(PPh 3 ) 4 (126) Mg, 0.11 mmol). The reaction mixture was heated at 90-95 ° C for 24 hours under nitrogen. Then 1 M NaOH (quantity) was added and heating was continued for another 2 hours. After cooling to room temperature, the reaction mixture was filtered, and then filtered, ethyl acetate was evaporated, filtered, concentrated, and acidified with 2 N HCl (pH 5-6). A precipitate was formed which was collected by suction and dried to give titled compound (yield: </RTI><RTIgt;
步驟4：N－(2－胺基－苯基)－4－{[4－(6－甲硫基－吡啶－3－基)－嘧啶－2－基胺基]－甲基}－苯甲醯胺(53) 依照與實例2步驟4(流程2)相同之程序自52以一步獲得呈乳白色固體之標題化合物53(實例45)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.60(s,1H),9.12(d,J＝1.8 Hz,1H),8.39(d,J＝5.3 Hz 1H),8.29(bd,J＝8.0 Hz,1H),7.98(t,J＝6.4 Hz,1H),7.94(d,J＝8.2 Hz,2H),7.56－7.38(m,3H),7.24(d,J＝5.1 Hz,1H),7.17(d,J＝7.0 Hz,1H),6.98(td,J＝7.5,1.4 Hz,1H),6.79(dd,J＝7.9,1.3 Hz 1H),6.60(td,J＝7.2,1.2 Hz,1H),4.90(s,2H),4.67(bd,J＝6.3 Hz,2H),2.60(s,3H)。 Step 4: N-(2-Amino-phenyl)-4-{[4-(6-methylthio-pyridin-3-yl)-pyrimidin-2-ylamino]-methyl}-benzene Amides (53) in accordance with example 2, step 4 (Scheme 2) the same procedure from the step 52 to obtain a milky white solid of the title compound 53 (example 45). 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.60 (s, 1H), 9.12 (d, J = 1.8 Hz, 1H), 8.39 (d, J = 5.3 Hz 1H), 8.29 (bd) , J=8.0 Hz, 1H), 7.98 (t, J=6.4 Hz, 1H), 7.94 (d, J=8.2 Hz, 2H), 7.56-7.38 (m, 3H), 7.24 (d, J=5.1 Hz) , 1H), 7.17 (d, J = 7.0 Hz, 1H), 6.98 (td, J = 7.5, 1.4 Hz, 1H), 6.79 (dd, J = 7.9, 1.3 Hz 1H), 6.60 (td, J = 7.2 , 1.2 Hz, 1H), 4.90 (s, 2H), 4.67 (bd, J = 6.3 Hz, 2H), 2.60 (s, 3H).
N－(2－胺基－苯基)－4－({4－[6－(1－羥基－1－甲基－乙基)－吡啶－3－基]－嘧啶－2－基胺基}－甲基)－苯甲醯胺(58a)步驟1：2－(5－溴－吡啶－2－基)－丙－2－醇(54a) 在－78℃下於氮氣下向2,5－二溴吡啶(2.00 g，8.44 mmol)之無水甲苯(100 mL)經攪拌溶液中緩緩加入正丁基鋰溶液(4.05 mL，10.13 mmol，己烷中2.5 M)。在－78℃下2小時後加入丙酮(806μl，10.98 mmol)。攪拌1小時後，使反應混合物溫至0℃且以飽和NH4 Cl中止之。形成一兩相系統；分離該有機層，以飽和NH4 Cl、H2 O及鹽水洗滌，將其經無水MgSO4 乾燥，過濾及濃縮。由急驟層析法經矽膠(AcOEt/CH2 Cl2 ：20/80)純化該殘餘物產生呈淺黃色含油液體之標題化合物54(1.37 g，6.34 mmol，產率75%)。1 H NMR(400 MHz,CDCl3 )δ(ppm)：ABX系統(δA ＝7.32,δB ＝7.82,δX ＝8.58,JA B ＝ 8.4 Hz,JB X ＝2.3 Hz,JA X ＝0 Hz,3H),4.47(bs,1H),1.57(s,6H)。 N-(2-Amino-phenyl)-4-({4-[6-(1-hydroxy-1-methyl-ethyl)-pyridin-3-yl]-pyrimidin-2-ylamino} -Methyl)-benzamide (58a) Step 1: 2-(5-Bromo-pyridin-2-yl)-propan-2-ol (54a) at -78 ° C under nitrogen to 2,5- Dibromopyridine (2.00 g, 8.44 mmol) of anhydrous toluene (100 mL) was slowly added to a solution of n-butyllithium (4.05 mL, 10.13 mmol, 2.5 M in hexane). Acetone (806 μl, 10.98 mmol) was added after 2 hours at -78 °C. After stirring for 1 hour, the reaction mixture was warmed to 0 ° C and quenched with saturated NH 4 Cl. Forming a two-phase system; the organic layer was separated, washed with saturated NH 4 Cl, H 2 O and brine, which was dried over anhydrous MgSO 4, filtered and concentrated. By flash chromatography over silica gel (AcOEt / CH 2 Cl 2: 20/80) The residue was purified to give the title compound as a pale yellow oily liquid of 54 (1.37 g, 6.34 mmol, 75% yield). 1 H NMR (400 MHz, CDCl 3 ) δ (ppm): ABX system (δ A = 7.32, δ B = 7.82, δ X = 8.58, J A B = 8.4 Hz, J B X = 2.3 Hz, J A X =0 Hz, 3H), 4.47 (bs, 1H), 1.57 (s, 6H).
步驟2：5－溴－2－(1－甲氧基甲氧基－1－甲基－乙基)－吡啶：(55) 在0℃於氮氣下向54(1.36 g，6.29 mmol)及i－Pr2 Net(2.19 mL，12.59 mmol)之無水二氯甲烷(20 mL)經攪拌溶液中緩緩加入氯甲基甲基醚(1.17 mL，14.63 mmol)。30分鐘後，將反應混合物溫至室溫，攪拌兩天，將其濃縮且由急驟層析法經矽膠(AcOEt/己烷：5/95)純化以產生呈淺黃色含油液體之標題化合物55(1.56 g，6.00 mmol，產率95%)。1 H NMR(400 MHz,CDCl3 )δ(ppm)：ABX系統(δA ＝7.50,δB ＝7.80,δX ＝8.60,JA B ＝8.4 Hz,JB X ＝2.4 Hz,JA X ＝0.8 Hz,3H),4.73(s,2H),3.41(s,3H),1.64(s,6H)。 Step 2: 5-Bromo-2-(1-methoxymethoxy-1-methyl-ethyl)-pyridine: (55) at 0 ° C under nitrogen to 54 (1.36 g, 6.29 mmol) and i -Pr 2 Net (2.19 mL, 12.59 mmol) in dry dichloromethane (20 mL)EtOAc. After 30 minutes, the reaction mixture was warmed to EtOAc EtOAc (mjqqqqqq 1.56 g, 6.00 mmol, yield 95%). 1 H NMR (400 MHz, CDCl 3 ) δ (ppm): ABX system (δ A = 7.50, δ B = 7.80, δ X = 8.60, J A B = 8.4 Hz, J B X = 2.4 Hz, J A X = 0.8 Hz, 3H), 4.73 (s, 2H), 3.41 (s, 3H), 1.64 (s, 6H).
步驟3：5－(2－[1－甲氧基甲氧基－1－甲基－乙基]－吡啶)－西朋酸(56) 在－50℃於氮氣下向溶於無水甲苯/THF(20 mL/5 mL)混合物中之55(2.90 g，11.15 mmol)及硼酸三異丙酯(3.09 mL，13.38 mmol)經攪拌溶液中經10分鐘逐滴加入正丁基鋰溶液(7.87 mL，13.38 mmol，己烷中1.7 M)。在50℃下45分鐘後，將混合物溫至室溫且在－30℃下以2 N HCl(30 mL)中止。傾析後，以1 N NaOH將含水層之pH調整至7，且將其以AcOEt萃取。蒸發萃取物且真空乾燥該殘餘物產生呈米色黏稠發泡體之標題化合物56a(2.33 g，10.37 mmol，產率93%)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：ABX系統(δA ＝7.55,δB ＝8.11,δX ＝8.83,JA B ＝7.8 Hz,JB X ＝1.8 Hz,JA X ＝0.9 Hz,3H),8.32(s,2H),4.68(s,2H),3.31(s,3H),1.56(s,6H)。 Step 3: 5-(2-[1-Methoxymethoxy-1-methyl-ethyl]-pyridine)-siponic acid (56) was dissolved in anhydrous toluene/THF under nitrogen at -50 °C. (20 mL/5 mL) 55 (2.90 g, 11.15 mmol) and triisopropyl borate (3.09 mL, 13.38 mmol) were added dropwise to a solution of n-butyllithium (7.87 mL) over 10 min. 13.38 mmol, 1.7 M in hexane. After 45 minutes at 50 °C, the mixture was warmed to room temperature and was quenched with 2 N HCl (30 mL) at -30 °C. After decantation, the pH of the aqueous layer was adjusted to 7 with 1 N NaOH and extracted with AcOEt. The extract was evaporated and dried <RTI ID=0.0> 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): ABX system (δ A = 7.55, δ B = 8.11, δ X = 8.83, J A B = 7.8 Hz, J B X = 1.8 Hz, J A X = 0.9 Hz, 3H), 8.32 (s, 2H), 4.68 (s, 2H), 3.31 (s, 3H), 1.56 (s, 6H).
步驟4：4－({4－[6－(1－羥基－1－甲基－乙基)－吡啶－3－基]－嘧啶－2－基胺基}－甲基)－苯甲酸(57a) 在室溫下向56(550 mg，2.44 mmol)、45b(510 mg，1.84 mmol，不純)及Na2 CO3 水溶液(10 mL，0.4 M)之混合物的乙腈(15 mL)經攪拌脫氣懸浮液中加入Pd(PPh3 )4 (106 mg，0.09 mmol)。氮氣下將反應混合物在95－100℃下加熱24小時，冷卻至室溫，過濾之，將濾液濃縮，以水稀釋，以AcOEt洗滌，以2 N HCl(25 mL)酸化且在60℃下溫4小時。將該反應混合物冷卻至室溫且以2 N NaOH將pH調整至5－6。形成沉澱物，其經過濾收集，以水沖洗且乾燥產生呈淺黃色固體之標題化合物57a(303 mg，0.83 mmol，產率45%)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：12.82(bs,1H),9.20－9.08(m,1H),8.46－8.32(m,2H,包括於8.40 ppm,d,J＝5.1 Hz),7.98(t,J＝6.2 Hz,1H),7.90(d,J＝8.2 Hz,2H),7.84－7.72(m,1H),7.58－7.42(m,2H),7.25(d,J＝5.1 Hz,1H),5.34(s,1H),4.67(d,J＝5.7 Hz,2H),1.50(s,6H)。 Step 4: 4-({4-[6-(1-Hydroxy-1-methyl-ethyl)-pyridin-3-yl]-pyrimidin-2-ylamino}-methyl)-benzoic acid (57a in acetonitrile) to 56 (550 mg, 2.44 mmol) , 45b (510 mg, 1.84 mmol, impure at room temperature) and Na 2 CO 3 solution (10 mL, 0.4 M) of (15 mL) was stirred degassed Pd(PPh 3 ) 4 (106 mg, 0.09 mmol) was added to the suspension. The reaction mixture was heated at 95-100 ° C for 24 hours under nitrogen, cooled to room temperature, filtered, and the filtrate was concentrated, diluted with water, washed with AcOEt, acidified with 2 N HCl (25 mL) and warm at 60 ° C 4 hours. The reaction mixture was cooled to room temperature and the pH was adjusted to 5-6 with 2 N NaOH. Precipitate was formed which was collected by EtOAc (EtOAc m.) 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 12.82 (bs, 1H), 9.20-9.08 (m, 1H), 8.46-8.32 (m, 2H, included in 8.40 ppm, d, J = 5.1 Hz), 7.98 (t, J = 6.2 Hz, 1H), 7.90 (d, J = 8.2 Hz, 2H), 7.84 - 7.72 (m, 1H), 7.58 - 7.42 (m, 2H), 7.25 (d, J = 5.1 Hz, 1H), 5.34 (s, 1H), 4.67 (d, J = 5.7 Hz, 2H), 1.50 (s, 6H).
步驟5：N－(2－胺基－苯基)－4－({4－[6－(1－羥基－1－甲基－乙基)－吡啶－3－基]－嘧啶－2－基胺基}－甲基)－苯甲醯胺(58a) 依照與實例2步驟4(流程2)相同之程序自57a以一步獲得呈乳白色固體之標題化合物58a(實例46)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.60(s,1H),9.15(d,J＝2.0 Hz,1H),8.45－8.35(m,2H,包括於8.41 ppm,d,J＝5.1 Hz),8.00(t,J＝6.4 Hz,1H),7.94(d,J＝8.2 Hz,2H),7.79(bd,J＝8.2 Hz,1H),7.58－7.43(m,2H),7.25(d,J＝5.1 Hz,1H),7.16(d,J＝7.4 Hz,1H),6.98(td,J＝7.6,1.5 Hz,1H),6.78(dd,J＝8.0,1.4 Hz 1H),6.60(t,J＝7.4 Hz,1H),5.37(s,1H),4.90(s,2H),4.68(d,J＝5.7 Hz,2H),1.50(s,6H)。 Step 5: N-(2-Amino-phenyl)-4-({4-[6-(1-hydroxy-1-methyl-ethyl)-pyridin-3-yl]-pyrimidin-2-yl Amino}-methyl)-benzamide (58a) The title compound 58a (yield 46) was obtained as a white solid from one step from 57a. 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.60 (s, 1H), 9.15 (d, J = 2.0 Hz, 1H), 8.45 - 8.35 (m, 2H, in 8.41 ppm, d , J=5.1 Hz), 8.00 (t, J=6.4 Hz, 1H), 7.94 (d, J=8.2 Hz, 2H), 7.79 (bd, J=8.2 Hz, 1H), 7.58-7.43 (m, 2H) ), 7.25 (d, J = 5.1 Hz, 1H), 7.16 (d, J = 7.4 Hz, 1H), 6.98 (td, J = 7.6, 1.5 Hz, 1H), 6.78 (dd, J = 8.0, 1.4 Hz) 1H), 6.60 (t, J = 7.4 Hz, 1H), 5.37 (s, 1H), 4.90 (s, 2H), 4.68 (d, J = 5.7 Hz, 2H), 1.50 (s, 6H).
N－(2－胺基－苯基)－4－({4－[6－(1－甲氧基甲氧基－1－甲基－乙基)－吡啶－3－基]－嘧啶－2－基胺基}－甲基)－苯甲醯胺(58b) 依照與實例46步驟4(注意：60℃下無酸水解)及5(流程15)相同之程序自56a獲得呈乳白色固體之標題化合物58b(實例47)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.61(s,1H),9.18(s,1H),8.50－8.32(m,2H,包括於8.42 ppm,d,J＝5.1 Hz),8.02(t,J＝6.0 Hz,1H),7.94(d,J＝8.0 Hz,2H),7.72(bd,J＝7.6 Hz,1H),7.59－7.42(m,2H),7.26(d,J＝5.1 Hz,1H),7.16(d,J＝7.8 Hz,1H),6.98(td,J＝7.6,1.4 Hz,1H),6.79(d,J＝7.8 Hz,1H),6.60(t,J＝7.4 Hz,1H),4.91(s,2H),4.72(s,2H),4.68(d,J＝5.3 Hz,2H),3.32(s,3H),1.60(s,6H)。 N-(2-Amino-phenyl)-4-({4-[6-(1-methoxymethoxy-1-methyl-ethyl)-pyridin-3-yl]-pyrimidine-2 -Aminomethyl}-methyl)-benzamide (58b) was obtained as a milky solid title from 56a according to the same procedure as in Example 46, Step 4 (Note: no acid hydrolysis at 60 ° C) and 5 (Scheme 15). Compound 58b (Example 47). 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.61 (s, 1H), 9.18 (s, 1H), 8.50-8.32 (m, 2H, included in 8.42 ppm, d, J = 5.1 Hz ), 8.02 (t, J = 6.0 Hz, 1H), 7.94 (d, J = 8.0 Hz, 2H), 7.72 (bd, J = 7.6 Hz, 1H), 7.59 - 7.42 (m, 2H), 7.26 (d) , J = 5.1 Hz, 1H), 7.16 (d, J = 7.8 Hz, 1H), 6.98 (td, J = 7.6, 1.4 Hz, 1H), 6.79 (d, J = 7.8 Hz, 1H), 6.60 (t , J = 7.4 Hz, 1H), 4.91 (s, 2H), 4.72 (s, 2H), 4.68 (d, J = 5.3 Hz, 2H), 3.32 (s, 3H), 1.60 (s, 6H).
N－(2－胺基－苯基)－4－[(2－甲硫基－6－吡啶－3－基－嘧啶－4－基胺基)－甲基]－苯甲醯胺(61a)步驟1：4－[(6－氯－2－甲硫基－嘧啶－4－基胺基)－甲基]－苯甲酸甲酯(59) 在70－80℃於氮氣下將溶於無水THF/DMF(10 mL/2 mL)之混合物中的4,6－二氯－2－(甲硫基)嘧啶(657 mg，3.37 mmol)或4,6－二氯－2－(R)－嘧啶、4－(胺甲基)苯甲酸甲酯氫氯酸鹽(744 mg，3.69 mmol)及i－Pr2 NEt(2.34 mL，13.43 mmol)經攪拌懸浮液加熱24小時。將混合物冷卻至室溫，傾入至飽和NaHCO3 且以AcOEt萃取之。以水、飽和NH4 Cl、H2 O及鹽水洗滌該有機層，將其經無水MgSO4 乾燥，過濾且進行濃縮。由急驟層析法經矽膠(溶離劑AcOEt/CH2 Cl2 ，30/70，接著40/60)純化該殘餘物產生呈米色粉末之標題化合物59a(929 mg，2.87 mmol，產率85%)。 N-(2-Amino-phenyl)-4-[(2-methylthio-6-pyridin-3-yl-pyrimidin-4-ylamino)-methyl]-benzamide (61a) Step 1: 4-[(6-Chloro-2-methylthio-pyrimidin-4-ylamino)-methyl]-benzoic acid methyl ester (59) is dissolved in anhydrous THF at 70-80 ° C under nitrogen. 4,6-Dichloro-2-(methylthio)pyrimidine (657 mg, 3.37 mmol) or 4,6-dichloro-2-(R)-pyrimidine in a mixture of /DMF (10 mL / 2 mL) Methyl 4-(aminomethyl)benzoate hydrochloride (744 mg, 3.69 mmol) and i-Pr 2 NEt (2.34 mL, 13.43 mmol) were stirred and stirred for 24 hours. The mixture was cooled to room temperature, poured into saturated NaHCO 3 and extracted them with AcOEt. With water, saturated NH 4 Cl, the organic layer was washed with H 2 O and brine, dried it over anhydrous MgSO 4, filtered and concentrated. By flash chromatography over silica gel (eluent AcOEt / CH 2 Cl 2, 30 /70, followed by 40/60) to give the residue as a beige powder of the title compound 59a (929 mg, 2.87 mmol, yield 85%) .
步驟2：4－[(2－甲硫基－6－吡啶－3－基－嘧啶－4－基胺基)－甲基]－苯甲酸(60) 室溫下向59(925 mg，2.86 mmol)、2,4,6－(3－吡啶基)－環三硼氧烷(360 mg，1.14 mmol)及Na2 CO3 水溶液(20 mL，0.4 M)之混合物的乙腈(20 mL)經攪拌脫氣懸浮液中加入Pd(PPh3 )4 (165 mg，0.14 mmol)。在95℃於氮氣下加熱該反應混合物一至兩天。將1 M NaOH(5 mL)加入反應混合物且再保持加熱1小時。將混合物冷卻至室溫且加以過濾。以AcOEt萃取濾液且將該含水層分離、濃縮，且以2 N HCl酸化(pH為5－7)。形成沉澱物，其經過濾收集及乾燥產生呈米色固體之標題化合物60(770 mg，2.19 mmol，產率76%)。1 H NMR(400 MHz,DMSO－d3 )δ(ppm)：12.89(bs 1H),9.15(bs,1H),8.72－8.64(m,1H),8.39－8.20(m,2H),7.93(d,J＝8.4 Hz,2H),7.58－7.40(m,3H),6.84(s,1H),4.69(d,J＝5.1 Hz2H),2.48(bs,3H)。 Step 2: 4-[(2-Methylthio-6-pyridin-3-yl-pyrimidin-4-ylamino)-methyl]-benzoic acid (60) at room temperature to 59 (925 mg, 2.86 mmol , a mixture of 2,4,6-(3-pyridyl)-cyclotriborane (360 mg, 1.14 mmol) and aq. Na 2 CO 3 (20 mL, 0.4 M) in EtOAc (20 mL) Pd(PPh 3 ) 4 (165 mg, 0.14 mmol) was added to the degassed suspension. The reaction mixture was heated at 95 ° C under nitrogen for one to two days. 1 M NaOH (5 mL) was added to the reaction mixture and kept warm for an additional 1 hour. The mixture was cooled to room temperature and filtered. The filtrate was extracted with AcOEt and the aqueous layer was separated, concentrated and acidified (pH 5-7) with 2 N HCl. Precipitate was formed which was filtered and dried to give the title compound 60 (yield: 770 mg, 2.19 mmol, yield 76%). 1 H NMR (400 MHz, DMSO-d 3 ) δ (ppm): 12.89 (bs 1H), 9.15 (bs, 1H), 8.72-8.64 (m, 1H), 8.39-8.20 (m, 2H), 7.93 ( d, J = 8.4 Hz, 2H), 7.58-7.40 (m, 3H), 6.84 (s, 1H), 4.69 (d, J = 5.1 Hz 2H), 2.48 (bs, 3H).
步驟3：N－(2－胺基－苯基)－4－[(2－甲硫基－6－吡啶－3－基－嘧啶－4－基胺基)－甲基]－苯甲醯胺(61a) 依照與實例2步驟4(流程2)相同之程序自60 以一步獲得呈乳白色固體之標題化合物61a(實例48)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.64(S,1H),9.16(bs,1H),8.68(dd,J＝4.7,1.6 Hz 1H),8.45－8.25(m,1H),8.27(t,J＝5.9 Hz,1H),7.97(d,J＝8.2 Hz,2H),7.58－7.42(m,3H),7.17(d,J＝7.6 Hz,1H),6.98(td,J＝7.5,1.4 Hz,1H),6.85(s,1H),6.79(dd,J＝7.9,1.1 Hz 1H),6.61(t,J＝7.2 Hz,1H),4.92(s,2H),4.69(bs,2H),2.50(s,3H)。 Step 3: N-(2-Amino-phenyl)-4-[(2-methylthio-6-pyridin-3-yl-pyrimidin-4-ylamino)-methyl]-benzamide (61a) The title compound 61a (yield 48) was obtained as a white solid from one step from 60 to the same procedure as in Example 2 Step 4 (Scheme 2). 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.64 (S, 1H), 9.16 (bs, 1H), 8.68 (dd, J = 4.7, 1.6 Hz 1H), 8.45-8.25 (m, 1H), 8.27 (t, J = 5.9 Hz, 1H), 7.97 (d, J = 8.2 Hz, 2H), 7.58-7.42 (m, 3H), 7.17 (d, J = 7.6 Hz, 1H), 6.98 ( Td, J = 7.5, 1.4 Hz, 1H), 6.85 (s, 1H), 6.79 (dd, J = 7.9, 1.1 Hz 1H), 6.61 (t, J = 7.2 Hz, 1H), 4.92 (s, 2H) , 4.69 (bs, 2H), 2.50 (s, 3H).
使用與對化合物61a所述(實例48，流程16)相同之程序製備實例49、50(化合物61b－c)。Examples 49, 50 (compounds 61b-c) were prepared using the same procedure as described for compound 61a (Example 48, Scheme 16).
N－(2－胺基－苯基)－4－[(6－吡啶－3－基－吡嗪－2－基胺基)－甲基]－苯甲醯胺(63)步驟1：4－[(6－氯－吡嗪－2－基胺基)－甲基]－苯甲酸甲酯(62) 在90℃於氮氣下將溶於無水THF/DMF(10 mL/2 mL)混合物中的2,6－二氯吡嗪(500 mg，3.36 mmol)、4－(胺甲基)苯甲酸甲酯氫氯酸鹽(744 mg，3.69 mmol)及i－Pr2 Net(2.05 mL，11.75 mmol)之經攪拌溶液加熱24小時。將混合物冷卻至室溫，傾入至NH4 Cl飽和水溶液中且以AcOEt萃取之。以H2 O及鹽水洗滌該有機層，使其經無水MgSO4 乾燥，過濾及乾燥。由急驟層析法經矽膠(AcOEt/CH2 Cl2 ：20/80→30/70)上純化該殘餘物產生呈淺黃色固體之標題化合物62(300 mg，1.08 mmol，產率32%)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：8.18(t,J＝6.0 Hz,1H),7.97(s,1H),AB系統(δA ＝7.95,δB ＝7.49,JA B ＝8.5 Hz,4H),7.75(s,1H),4.58(d,J＝5.9 Hz,2H),3.87(s,3H)。 N-(2-Amino-phenyl)-4-[(6-pyridin-3-yl-pyrazin-2-ylamino)-methyl]-benzamide (63) Step 1: 4- [(6-Chloro-pyrazin-2-ylamino)-methyl]-benzoic acid methyl ester (62) was dissolved in anhydrous THF / DMF (10 mL / 2 mL) mixture at 90 ° C under nitrogen. 2,6-dichloropyrazine (500 mg, 3.36 mmol), methyl 4-(aminomethyl)benzoate hydrochloride (744 mg, 3.69 mmol) and i-Pr 2 Net (2.05 mL, 11.75 mmol The stirred solution was heated for 24 hours. The mixture was cooled to room temperature, poured into saturated aqueous NH 4 Cl and extracted with AcOEt in it. Washed with H 2 O and brine, the organic layer was dried over anhydrous MgSO 4, filtered and dried. Over silica gel (AcOEt / CH 2 Cl 2: 20/80 → 30/70) was purified by flash chromatography on the residue produced the title compound as a pale yellow solid 62 (300 mg, 1.08 mmol, 32% yield). 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 8.18 (t, J = 6.0 Hz, 1H), 7.97 (s, 1H), AB system (δ A = 7.95, δ B = 7.49, J A B = 8.5 Hz, 4H), 7.75 (s, 1H), 4.58 (d, J = 5.9 Hz, 2H), 3.87 (s, 3H).
步驟2：N－(2－胺基－苯基)－4－[(6－吡啶－3－基－吡嗪－2－基胺基)－甲基]－苯甲醯胺(63) 依照與實例48步驟2及3(流程16)相同之程序自62以兩步獲得呈米色粉末之標題化合物63(實例51)。1 H NMR(400 MHz,DMSO－d6 )δ(ppm)：9.62(s,1H),9.19(dd,J＝2.3,0.8 Hz,1H),8.62(dd,J＝4.7,1.6 Hz 1H),8.40(s,1H),8.36(ddd,J＝8.4,2.0,2.0 Hz,1H),8.06(s,1H),8.01－7.92(m,3H),7.54(d,J＝8.2 Hz,2H),7.51(ddd,J＝7.8,4.7,0.8 Hz,1H),7.16(bd,J＝6.7 Hz,1H),6.98(td,J＝7.6,1.5 Hz,1H),6.78(dd,J＝8.0,1.4 Hz 1H),6.60(td,J＝7.5,1.3 Hz,1H),4.91(s,2H),4.71(d,J＝6.1 Hz,2H)。 Step 2: N-(2-Amino-phenyl)-4-[(6-pyridin-3-yl-pyrazin-2-ylamino)-methyl]-benzamide (63) according to Example 48 Steps 2 and 3 (Scheme 16) The same procedure was used to obtain the title compound 63 (Example 51) as a beige powder from 62. 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm): 9.62 (s, 1H), 9.19 (dd, J = 2.3, 0.8 Hz, 1H), 8.62 (dd, J = 4.7, 1.6 Hz 1H) , 8.40 (s, 1H), 8.36 (ddd, J = 8.4, 2.0, 2.0 Hz, 1H), 8.06 (s, 1H), 8.01-7.92 (m, 3H), 7.54 (d, J = 8.2 Hz, 2H ), 7.51 (ddd, J = 7.8, 4.7, 0.8 Hz, 1H), 7.16 (bd, J = 6.7 Hz, 1H), 6.98 (td, J = 7.6, 1.5 Hz, 1H), 6.78 (dd, J = 8.0, 1.4 Hz 1H), 6.60 (td, J = 7.5, 1.3 Hz, 1H), 4.91 (s, 2H), 4.71 (d, J = 6.1 Hz, 2H).
N－(2－胺基－苯基)－4－[(6－嗎啉－4－基－噠嗪－3－基胺基)－甲基]－苯甲醯胺(66)步驟1：4－(6－氯－噠嗪－3－基)－嗎啉(64) 向一配有一回流冷凝器之50 mL燒瓶中饋入嗎啉(2.93 mL，33.5 mmol)及3,6－二氯噠嗪(5.00 g，33.5 mmol)。將混合物在90℃下加熱6小時，將所得固體在EtOAc、水與飽和NH4 Cl之間分溶。分離有機層，將其經無水Na2 SO4 乾燥，過濾及蒸發產生標題化合物64(5.3 g，26.5 mmol，產率79%)。 N-(2-Amino-phenyl)-4-[(6-morpholin-4-yl-pyridazin-3-ylamino)-methyl]-benzamide (66) Step 1:4 -(6-Chloro-pyridazin-3-yl)-morpholine (64) was fed a morpholine (2.93 mL, 33.5 mmol) and 3,6-dichloropurine in a 50 mL flask equipped with a reflux condenser. Azine (5.00 g, 33.5 mmol). The mixture was heated for 6 hours at 90 ℃, and the resulting solid was partitioned between EtOAc, water and saturated NH 4 Cl. The organic layer was separated, dried over anhydrous Na 2 SO 4, filtered, and evaporated to give the title compound 64 (5.3 g, 26.5 mmol, 79% yield).
步驟2：4－[(6－嗎啉－4－基－噠嗪－3－基胺基)－甲基]－苯甲酸甲酯(65) 向64(2.0 g，10.0 mmol)及4－(胺甲基)苯甲酸甲酯氫氯酸鹽(2.2 g，11.0 mmol)之異丙二醇(200 mL)溶液中加入NH4 Cl(2.14 g，40.0 mmol)。將反應混合物在150℃下加熱72小時且加以濃縮。將殘餘物溶解於水中且以萃取。分離含水相，將其以1 N NaOH(pH 8)處理且以EtOAc萃取之。將該萃取物經無水Na2 SO4 乾燥、過濾及濃縮。由急驟層析法經矽膠(MeOH/CH2 Cl2 ：2/98至5/95)純化該殘餘物產生標題化合物65(270 mg，0.82 mmol，產率8%)。1 H NMR：(400 MHz,CDCl3 )δ(ppm)：7.97(d,J＝8.0 Hz,2H),7.42(d,J＝8.0 Hz,2H),6.90(d,J＝9.2 Hz,1H),6.69(d,J＝9.2 Hz,1H),5.16－5.06(bs,1H),4.67(s,2H),3.90(s,3H),3.82(t,J＝4.8 Hz,4H),3.41(t,J＝4.8 Hz,4H)。 Step 2: 4-[(6-morpholin-4-yl-pyridazin-3-ylamino)-methyl]-benzoic acid methyl ester (65) to 64 (2.0 g, 10.0 mmol) and 4- ( aminomethyl) benzoate hydrochloride (2.2 g, 11.0 mmol) of isopropylene glycol (200 mL) was added NH 4 Cl (2.14 g, 40.0 mmol). The reaction mixture was heated at 150 ° C for 72 hours and concentrated. The residue was dissolved in water and extracted. The aqueous phase was separated, treated with 1 N NaOH (pH 8) and extracted with EtOAc. The extract was dried over anhydrous Na 2 SO 4, filtered and concentrated. (MeOH / CH 2 Cl 2: 2/98 to 5/95) over silica gel by flash chromatography of the residue was purified to give the title compound 65 (270 mg, 0.82 mmol, 8% yield). 1 H NMR: (400 MHz, CDCl 3 ) δ (ppm): 7.97 (d, J = 8.0 Hz, 2H), 7.42 (d, J = 8.0 Hz, 2H), 6.90 (d, J = 9.2 Hz, 1H) ), 6.69 (d, J = 9.2 Hz, 1H), 5.16-5.06 (bs, 1H), 4.67 (s, 2H), 3.90 (s, 3H), 3.82 (t, J = 4.8 Hz, 4H), 3.41 (t, J = 4.8 Hz, 4H).
步驟3：N－(2－胺基－苯基)－4－[(6－嗎啉－4－基－噠嗪－3－基胺基)－甲基]－苯甲醯胺(66) 依照與實例2步驟3及4(流程2)相同之程序自65以兩步獲得呈乳白色固體之標題化合物66(實例52)。1 H NMR：(400 MHz,DMSO－d6 )δ(ppm)：9.57(s,1H),7.89(d,J＝8.0 Hz,2H),7.42(d,J＝8.0 Hz,2H),7.15－7.12(m,1H),7.13(d,J＝9.6 Hz,1H),6.94(t,J＝7.6 Hz,1H),6.88(t,J＝6.0 Hz,1H),6.83(d,J＝9.6 Hz,1H),6.75(dd,J＝8.0,1.2 Hz,1H),6.57(t,J＝8.0 Hz,1H),4.86(s,2H),4.54(d,J＝6.4 Hz,2H),3.69(t,J＝4.8 Hz,4H),3.24(t,J＝4.8 Hz,4H)。 Step 3: N-(2-Amino-phenyl)-4-[(6-morpholin-4-yl-pyridazin-3-ylamino)-methyl]-benzamide (66) The same procedure as Example 2, Steps 3 and 4 (Scheme 2) was obtained from the title compound 66 (Example 52) as a white solid. 1 H NMR: (400 MHz, DMSO-d 6 ) δ (ppm): 9.57 (s, 1H), 7.89 (d, J = 8.0 Hz, 2H), 7.42 (d, J = 8.0 Hz, 2H), 7.15 -7.12(m,1H),7.13(d,J=9.6 Hz,1H), 6.94(t,J=7.6 Hz,1H),6.88(t,J=6.0 Hz,1H),6.83(d,J= 9.6 Hz, 1H), 6.75 (dd, J=8.0, 1.2 Hz, 1H), 6.57 (t, J = 8.0 Hz, 1H), 4.86 (s, 2H), 4.54 (d, J = 6.4 Hz, 2H) , 3.69 (t, J = 4.8 Hz, 4H), 3.24 (t, J = 4.8 Hz, 4H).
N－(2－胺基－苯基)－4－[2－酮基－2－(4－嘧啶－2－基－六氫吡嗪－1－基)－乙基]－苯甲醯胺(71)步驟1：4－氰甲基－苯甲酸(68) 根據J.Med.Chem.1997,40,377－384描述之程序自4－氯甲基苯甲酸(67)開始獲得標題化合物。 N-(2-Amino-phenyl)-4-[2-keto-2-(4-pyrimidin-2-yl-hexahydropyrazin-1-yl)-ethyl]-benzamide ( 71) Step 1: 4-Cyanomethyl-benzoic acid (68) The title compound was obtained starting from 4-chloromethylbenzoic acid (67) according to the procedure described in J. Med. Chem. 1997, 40, 377-384.
步驟2：4－羧甲基－苯甲酸甲酯(69) 根據J.Med.Chem.1998,41,5219－5246描述之程序獲得呈米色固體之標題化合物(產率85%)。1 H NMR (DMSO－d6 )δ(ppm)：12.49(s,1H),7.89(d,J＝1.8 Hz,2H),7.39(d,J＝8.2 Hz,2H),3.84(s,3H),3.79(s,2H)。 Step 2: 4-Carboxymethyl-benzoic acid methyl ester (69) The title compound (yield: 85%) was obtained as a beige solid, according to the procedure described in J. Med. Chem. 1998, 41, 5219-5246. 1 H NMR (DMSO-d 6 ) δ (ppm): 12.49 (s, 1H), 7.89 (d, J = 1.8 Hz, 2H), 7.39 (d, J = 8.2 Hz, 2H), 3.84 (s, 3H) ), 3.79 (s, 2H).
步驟3：4－[2－酮基－2－(4－吡啶－2－基－六氫吡嗪－1－基)－乙基]－苯甲酸甲酯(70) 依照實例1步驟5(流程1)描述之程序獲得呈淺黃色固體之標題化合物70 (產率70%)。1 H－NMR(DMSO)δ：8.36(d,J＝4.7 Hz,2H),7.89(d,J＝8.4 Hz,2H),7.38(d,J＝8.4 Hz,2H),6.65(t,J＝4.8 Hz,1H),3.87(s,2H),3.83(s,3H),3.71－3.66(m,4H),3.60－3.53(m,4H)。 Step 3: 4-[2-keto-2-(4-pyridin-2-yl-hexahydropyrazin-1-yl)-ethyl]-benzoic acid methyl ester (70) according to Example 1 Step 5 (Process 1) Procedure to give the title compound 70 (yield 70%) as a pale yellow solid. 1 H-NMR (DMSO) δ: 8.36 (d, J = 4.7 Hz, 2H), 7.89 (d, J = 8.4 Hz, 2H), 7.38 (d, J = 8.4 Hz, 2H), 6.65 (t, J) = 4.8 Hz, 1H), 3.87 (s, 2H), 3.83 (s, 3H), 3.71-3.66 (m, 4H), 3.60-3.53 (m, 4H).
步驟4：N－(2－胺基－苯基)－4－[2－酮基－2－(4－嘧啶－2－基－六氫吡嗪－1－基)－乙基]－苯甲醯胺(71) 依照實例1步驟4及5描述之程序獲得呈米色固體之標題化合物71(225 mg，69%)。1 H NMR：(DMSO)δ(ppm)：9.62(s,1H),8.36(d,J＝4.7 Hz,2H),7.90(d,J＝8.0 Hz,2H),7.36(d,J＝8.2 Hz,2H),7.13(d,J＝6.8 Hz,1H),6.95(td,J＝7.5,1.4 Hz,1H),6.76(dd,J＝8.0,1.4 Hz,1H),6.70(t,J＝4.8 Hz,1H),6.57(tdd,J＝7.6,1.4 Hz,1H),4.90(s,2H),3.87(s,2H),3.72－3.68(m,4H),3.62－3.55(m,4H)。 Step 4: N-(2-Amino-phenyl)-4-[2-keto-2-(4-pyrimidin-2-yl-hexahydropyrazin-1-yl)-ethyl]-benzene Amides (71) was obtained as an off-white solid of the title compound 71 (225 mg, 69%) according to the procedure described in step 4 of example 1 and 5. 1 H NMR: (DMSO) δ (ppm): 9.62 (s, 1H), 8.36 (d, J = 4.7 Hz, 2H), 7.90 (d, J = 8.0 Hz, 2H), 7.36 (d, J = 8.2 Hz, 2H), 7.13 (d, J = 6.8 Hz, 1H), 6.95 (td, J = 7.5, 1.4 Hz, 1H), 6.76 (dd, J = 8.0, 1.4 Hz, 1H), 6.70 (t, J = 4.8 Hz, 1H), 6.57 (tdd, J = 7.6, 1.4 Hz, 1H), 4.90 (s, 2H), 3.87 (s, 2H), 3.72-3.68 (m, 4H), 3.62-3.55 (m, 4H).
N－(2－胺基－苯基)－4－(5－氯－6－氟－1H－苯幷咪唑－2－基甲基)－苯甲醯胺(74)步驟1：4－[(2－胺基－4－氯－5－氟－苯胺甲醯基)－甲基]－苯甲酸甲酯(72) 依照實例1步驟5(流程1)描述之程序獲得呈橙色油狀物之標題化合物72(產率69%)。LRMS：336.1(計算值)，337.5(獲得值)。 N-(2-Amino-phenyl)-4-(5-chloro-6-fluoro-1H-benzimidazol-2-ylmethyl)-benzamide (74) Step 1: 4-[( 2-Amino-4-chloro-5-fluoro-anilinylcarbazyl)-methyl]-benzoic acid methyl ester (72) was obtained as an orange oily title according to the procedure described in Example 1 Step 5 (Scheme 1). Compound 72 (yield 69%). LRMS: 336.1 (calculated), 337.5 (obtained).
步驟2：4－(5－氯－6－氟－1H－苯幷咪唑－2－基甲基)－苯甲酸甲酯(73) 將化合物72(333 mg，0.99 mmol)溶解於AcOH(10 ml)中且使溶液回流24小時。蒸發AcOH且將該殘餘物溶解於AcOEt中，以水性NH4 Cl、NaHCO3 及鹽水洗滌，且經MgSO4 乾燥。蒸發EtOAc提供呈褐色粉末之標題化合物73(297 mg，95%)。 Step 2: 4-(5-Chloro-6-fluoro-1H-benzimidazol-2-ylmethyl)-benzoic acid methyl ester (73) Compound 72 (333 mg, 0.99 mmol) was dissolved in AcOH (10 ml And the solution was refluxed for 24 hours. AcOH was evaporated and the residue was dissolved in AcOEt, washed with aqueous NH 4 Cl, NaHCO 3 and brine, and dried over MgSO 4. The title compound 73 (297 mg, 95%)
1 H NMR(DMSO－d6 )δ(ppm)：7.90(d,J＝1.8 Hz,2H),7.88(d,J＝1.9 Hz,1H),7.66(d,J＝7 Hz,1H),7.51(d,J＝9.8 Hz,2H),4.26(s,2H),3.32(s,3H)。 1 H NMR (DMSO-d 6 ) δ (ppm): 7.90 (d, J = 1.8 Hz, 2H), 7.88 (d, J = 1.9 Hz, 1H), 7.66 (d, J = 7 Hz, 1H), 7.51 (d, J = 9.8 Hz, 2H), 4.26 (s, 2H), 3.32 (s, 3H).
LRMS：318.1(計算值)，319.4(獲得值)。LRMS: 318.1 (calculated), 319.4 (obtained).
步驟3：N－(2－胺基－苯基)－4－(5－氯－6－氟－1H－苯幷咪唑－2－基甲基)－苯甲醯胺(74) 依照實例1步驟4及5描述之程序獲得呈黃色固體之標題化合物74(產率53%)。1 H NMR：(DMSO)δ(ppm)：9.59(s,1H),7.91(d,J＝8.0 Hz,2H),7.67(d,J＝6.9 Hz,1H),7.51(d,J＝9.7 Hz,1H),7.43(d,J＝8.4 Hz,2H),7.13(d,J＝7.2 Hz,1H),6.74(td,J＝7.6 Hz,1H),7.52(dd,J＝4.7,4.7 Hz,1H),6.57(dd,J＝7.0,1.4 Hz,2H),4.87(s,2H),4.26(s,2H)。 Step 3: N-(2-Amino-phenyl)-4-(5-chloro-6-fluoro-1H-benzimidazol-2-ylmethyl)-benzamide (74) according to Example 1 The title compound 74 was obtained as a yellow solid (yield: 53%). 1 H NMR: (DMSO) δ (ppm): 9.59 (s, 1H), 7.91 (d, J = 8.0 Hz, 2H), 7.67 (d, J = 6.9 Hz, 1H), 7.51 (d, J = 9.7 Hz, 1H), 7.43 (d, J = 8.4 Hz, 2H), 7.13 (d, J = 7.2 Hz, 1H), 6.74 (td, J = 7.6 Hz, 1H), 7.52 (dd, J = 4.7, 4.7 Hz, 1H), 6.57 (dd, J = 7.0, 1.4 Hz, 2H), 4.87 (s, 2H), 4.26 (s, 2H).
N－(2－胺基－苯基)－4－(2,4－二酮基－1,4－二氫－2H－噻吩幷[2,3－d]嘧啶－3－基甲基)－苯甲醯胺(77) 依照與專利申請案WO 03/024448所述相同之程序自2－胺基－噻吩－3－甲酸甲酯開始經由中間體75及76(流程20)獲得標題化合物77。1 H NMR(DMSO)δ(ppm)：9.59(s,1H),7.88(d,J＝8.2 Hz,2H),7.38(d,J＝8.2 Hz,2H),7.18－7.11(m,3H),7.45(bs,1H),6.94(td,J＝7.6,1.6 Hz,1H),6.75(dd,J＝7.8,1.4 Hz,1H),6.57(td,J＝7.5,1.4 Hz,1H),5.09(s,2H),4.87(bs,2H)。LRMS：392.1(計算值)，393.4(獲得值)。 N-(2-Amino-phenyl)-4-(2,4-dione-1,4-dihydro-2H-thiophene[2,3-d]pyrimidin-3-ylmethyl)- Benzoguanamine (77) The title compound 77 was obtained via intermediates 75 and 76 (Scheme 20) starting from methyl 2-amino-thiophene-3-carboxylate starting from the same procedure as described in the patent application WO 03/024448. 1 H NMR (DMSO) δ (ppm): 9.59 (s, 1H), 7.88 (d, J = 8.2 Hz, 2H), 7.38 (d, J = 8.2 Hz, 2H), 7.18-7.11 (m, 3H) , 7.45 (bs, 1H), 6.94 (td, J = 7.6, 1.6 Hz, 1H), 6.75 (dd, J = 7.8, 1.4 Hz, 1H), 6.57 (td, J = 7.5, 1.4 Hz, 1H), 5.09 (s, 2H), 4.87 (bs, 2H). LRMS: 392.1 (calculated), 393.4 (obtained).
依照與專利申請案WO 03/024448所述相同之程序自3-胺基-噻吩-2-甲酸甲酯開始經由中間體78及79獲得呈黃色固體之標題化合物80。1 H NMR(DMSO)δ(ppm)：9.60(s,1H),8.22(d,J=5.5Hz,1H),7.89(d,J=8.2Hz,2H),7.41-7.39(m,3H),7.13(d,J=7.4Hz,1H),6.95(td,J=7.6,1.6Hz,1H),6.75(dd,J=7.8,1.6Hz,1H),6.57(td,J=7.4,1.2Hz,1H),5.16(s,2H),4.88(bs,2H),4.23(m,2H),2.81(m,2H)。LRMS：463.2(計算值)，464.4(獲得值)。The title compound 80 was obtained as a yellow solid from intermediates 78 and 79, starting from methyl 3-amino-thiophene-2-carboxylate, starting from the same procedure as described in the patent application WO 03/024448. 1 H NMR (DMSO) δ (ppm): 9.60 (s, 1H), 8.22 (d, J = 5.5 Hz, 1H), 7.89 (d, J = 8.2 Hz, 2H), 7.41 - 7.39 (m, 3H) , 7.13 (d, J = 7.4 Hz, 1H), 6.95 (td, J = 7.6, 1.6 Hz, 1H), 6.75 (dd, J = 7.8, 1.6 Hz, 1H), 6.57 (td, J = 7.4, 1.2 Hz, 1H), 5.16 (s, 2H), 4.88 (bs, 2H), 4.23 (m, 2H), 2.81 (m, 2H). LRMS: 463.2 (calculated), 464.4 (obtained).
{5－[4－(2－胺基－苯胺甲醯基)－苄基]－3－氰基－4－甲基－噻吩－2－基}－胺基甲酸2－嗎啉－4－基－乙酯(83)步驟1：4－[4－氰基－3－甲基－5－(2－嗎啉－4－基－乙氧羰基胺基)－噻吩－2－基甲基]－苯甲酸(82) 在5℃下向羰基二咪唑(207 mg，1.28 mM)之無水THF(10 ml)溶液中加入羥乙基嗎啉(114μl，1.28 mM)。移開冷卻，在室溫下攪拌該反應混合物1小時且在室溫下經由套管將其加入至4－(5－胺基－4－氰基－3－甲基－噻吩－2－基甲基)－苯甲酸(81，350 mg，1.28 mM)(描述於專利申請案WO 03/024448)及DBU(382μl，2.56 mM)之無水THF(20 ml)溶液中。攪拌該經組合之混合物3小時，蒸發THF且將剩餘之固體殘餘物懸浮於水中，以濃HCl(pH 4)酸化且加以過濾收集。以25 ml丙酮濕磨該材料提供標題化合物82(85 mg，產率15%)。 {5-[4-(2-Amino-anilinemethanyl)-benzyl]-3-cyano-4-methyl-thiophen-2-yl}-carbamic acid 2-morpholin-4-yl -ethyl ester (83) Step 1: 4-[4-Cyano-3-methyl-5-(2-morpholin-4-yl-ethoxycarbonylamino)-thiophen-2-ylmethyl]- Benzoic acid (82) To a solution of carbonyldiimidazole (207 mg, 1.28 mM) in dry THF (10 ml) was added hydroxyethylmorpholine (114 μl, 1.28 mM) at 5 °C. The cooling was removed, the reaction mixture was stirred at room temperature for 1 hour and added to the 4-(5-amino-4-cyano-3-methyl-thiophen-2-yl group via a cannula at room temperature Benzoic acid (81,350 mg, 1.28 mM) (described in patent application WO 03/024448) and DBU (382 μl, 2.56 mM) in anhydrous THF (20 ml). The combined mixture was stirred for 3 hours, THF was evaporated and the remaining solid residue was suspended in water, acidified with concentrated HCl (pH 4) and collected by filtration. The material was triturated with 25 ml of acetone to afford title compound 82 (85 mg, yield 15%).
LRMS：429.5(計算值)，430.4(實驗值)。LRMS: 429.5 (calculated), 430.4 (exp.).
步驟2：{5－[4－(2－胺基－苯胺甲醯基)－苄基]－3－氰基－4－甲基－噻吩－2－基}－胺基甲酸2－嗎啉－4－基－乙酯(83) 依照實例1步驟5描述之程序獲得呈固體之標題化合物83(產率26%)。1 H NMR：(300 MHz,DMSO－d6 ,δ(ppm)：9.61(s,1H),7.92(d,J＝7.91,2H),7.34(d,J＝7.91,2H),7.15(d,J＝7.47,1H),6.97(t,J＝7.03,1H),6.77(d,J＝7.03,1H),6.59(t,J＝7.47,1H),4.88(brs,2H),4.22(t,J＝5.50,2H),4.10(s,2H),3.55(t,J＝4.40,4H),2.56(t,J＝5.50,2H),2.43－2.39(m,4H),2.18(s,3H)。LRMS：519.6(計算值)，520.5(實驗值)。 Step 2: {5-[4-(2-Amino-anilinemethanyl)-benzyl]-3-cyano-4-methyl-thiophen-2-yl}-carbamic acid 2-morpholine- 4-Base-Ethyl Ester (83) The title compound (yield 26%) was obtained as a solid. 1 H NMR: (300 MHz, DMSO-d 6 , δ (ppm): 9.61 (s, 1H), 7.92 (d, J = 7.91, 2H), 7.34 (d, J = 7.91, 2H), 7.15 (d) , J = 7.47, 1H), 6.97 (t, J = 7.03, 1H), 6.77 (d, J = 7.03, 1H), 6.59 (t, J = 7.47, 1H), 4.88 (brs, 2H), 4.22 ( t, J = 5.50, 2H), 4.10 (s, 2H), 3.55 (t, J = 4.40, 4H), 2.56 (t, J = 5.50, 2H), 2.43 - 2.39 (m, 4H), 2.18 (s) , 3H). LRMS: 519.6 (calculated), 520.5 (exp.).
N－(2－胺基－苯基)－4－(4－氰基－5－丙醯胺基－噻吩－2－基甲基)－苯甲醯胺(87)步驟1：4－(3－酮基－丙基)－苯甲酸甲酯(84) 根據J.Org.Chem.；1992； 57(11)；3218－3225描述之程序自4－碘代苯甲酸甲酯開始獲得標題化合物84。 N-(2-Amino-phenyl)-4-(4-cyano-5-propionamido-thiophen-2-ylmethyl)-benzamide (87) Step 1: 4-(3) -Ketyl -propyl)-methyl benzoate (84) The title compound was obtained from the methyl 4-iodobenzoate according to the procedure described in J. Org. Chem.; 1992; 57(11); 3218-3225. .
步驟2：4－(5－胺基－4－氰基－噻吩－2－基甲基)－苯甲酸甲酯(85) 在0℃下向硫(309 mg，4.22 mmol)及4－(3－酮基－丙基)－苯甲酸甲酯(84)(812 mg，4.22 mmol)之DMF(4 ml)懸浮液中緩緩加入Et3 N(353μl，2.53 mmol)。在室溫下攪拌反應混合物1小時且緩緩添加丙二腈(279 mg，4.22 mmol)之DMF(6 ml)溶液。在室溫下攪拌反應混合物16小時，將其傾入至300 ml冰/水中以產生橙色沉澱物，以冷水沖洗及乾燥該沉澱物產生呈橙色固體之標題化合物85(1.04 g，90%)。LRMS：272.1(計算值)；265.0(實驗值)。 Step 2: 4-(5-Amino-4-cyano-thiophen-2-ylmethyl)-benzoic acid methyl ester (85) to sulfur (309 mg, 4.22 mmol) and 4-(3) - keto - propyl) - benzoic acid methyl ester (84) (812 mg, 4.22 mmol) of DMF (4 ml) was slowly added to a suspension of Et 3 N (353μl, 2.53 mmol ). The reaction mixture was stirred at room temperature for 1 hour and a solution of malononitrile (279 mg, 4.22 mmol) in DMF (6 ml). The reaction mixture was stirred at room temperature for 16 hrs, EtOAc (EtOAc m. LRMS: 272.1 (calculated); 265.0 (exp.).
步驟3，4：N－(2－胺基－苯基)－4－(4－氰基－5－丙醯胺基－噻吩－2－基甲基)－苯甲醯胺(87) 依照與專利申請案WO 03/024448關於類似化合物所述相同之程序自氰基化合物85開始經由中間體86獲得呈黃色固體之標題化合物87(產率63%)。1 H NMR(DMSO)δ(ppm)：11.26(s,1H),9.61(s,1H),7.92(d,J＝7.5 Hz,2H),7.39(d,J＝7.5 Hz,2H),7.15(d,J＝7.5 Hz,1H),6.95(s,2H),6.77(d,J＝8.0 Hz,1H),6.58(m,1H),5.76－5.75(m,1H),4.88(s,2H),4.12(s,2H),2.50(m,3H),1.05－1.03(m,3H)。LRMS：404.1(計算值),405.1(獲得值)。 Step 3, 4: N-(2-Amino-phenyl)-4-(4-cyano-5-propionamido-thiophen-2-ylmethyl)-benzamide (87) according to The title compound 87 (yield 63%) was obtained as a yellow solid from Intermediate s. 1 H NMR (DMSO) δ (ppm): 11.26 (s, 1H), 9.61 (s, 1H), 7.92 (d, J = 7.5 Hz, 2H), 7.39 (d, J = 7.5 Hz, 2H), 7.15 (d, J = 7.5 Hz, 1H), 6.95 (s, 2H), 6.77 (d, J = 8.0 Hz, 1H), 6.58 (m, 1H), 5.76-5.75 (m, 1H), 4.88 (s, 2H), 4.12 (s, 2H), 2.50 (m, 3H), 1.05-1.03 (m, 3H). LRMS: 404.1 (calculated), 405.1 (obtained).
{5－[4－(2－胺基－苯胺甲醯基)－苄基]－3－氰基－噻吩－2－基}－胺基甲酸2－嗎啉－4－基－乙酯(89) 類似於化合物87實例58(流程23)，自氰基化合物85開始經由中間體88獲得呈黃色固體之標題化合物89(產率15%)。1 H NMR：(300 MHz,DMSO－d6 ,δ(ppm)：9.60(bs,1H),7.91(d,J＝7.8 Hz,2H),7.38(d,J＝8.2 Hz,2H),7.13(d,J＝7.8 Hz,1H),6.97－6.93(m,2H),6.76(dd,J＝7.8,1.2 Hz,1H),6.58(ddd,J＝7.4,7.4,1.2 Hz,1H),4.89(bs,2H),4.22(t,J＝5.5 Hz,2H),4.12(s,2H),3.55(t,J＝4.7 Hz,4H),2.56(t,J＝5.7 Hz,2H),2.41(m,4H)。 {5-[4-(2-Amino-anilinomethyl)-benzyl]-3-cyano-thiophen-2-yl}-carbamic acid 2-morpholin-4-yl-ethyl ester (89 ) In analogy to example 58 compound 87 (Scheme 23), obtained starting from the cyano compound 85 the title compound 89 as a yellow solid (15% yield) of intermediate 88 via the. 1 H NMR: (300 MHz, DMSO-d 6 , δ (ppm): 9.60 (bs, 1H), 7.91 (d, J = 7.8 Hz, 2H), 7.38 (d, J = 8.2 Hz, 2H), 7.13 (d, J = 7.8 Hz, 1H), 6.97-6.93 (m, 2H), 6.76 (dd, J = 7.8, 1.2 Hz, 1H), 6.58 (ddd, J = 7.4, 7.4, 1.2 Hz, 1H), 4.89(bs, 2H), 4.22 (t, J = 5.5 Hz, 2H), 4.12 (s, 2H), 3.55 (t, J = 4.7 Hz, 4H), 2.56 (t, J = 5.7 Hz, 2H), 2.41 (m, 4H).
4－(5－胺基－4－氰基－噻吩－2－基甲基)－N－(2－胺基－苯基)－苯甲醯胺(90) 依照與實例1步驟4及5所述相同之程序自化合物85開始獲得呈橙色固體之標題化合物90(產率67%)。1 H NMR：(DMSO)δ(ppm)：9.60(bs,1H),7.90(d,J＝8.2 Hz,2H),7.33(d,J＝8.2 Hz,2H),7.13(d,J＝6.7 Hz,1H),7.03(bs,1H),6.95(ddd,J＝7.6,7.6,1.6 Hz,1H),6.76(dd,J＝6.6,1.6 Hz,1H),6.58(ddd,J＝7.4,7.4,1.7 Hz,1H),6.52(s,1H),4.89(bs,2H),3.95(s,2H)。LRMS：348.1(計算值)；349.2(實驗值)。 4-(5-Amino-4-cyano-thiophen-2-ylmethyl)-N-(2-amino-phenyl)-benzamide (90) according to steps 4 and 5 of Example 1. The title compound 90 (yield 67%) was obtained as an orange solid. 1 H NMR: (DMSO) δ (ppm): 9.60 (bs, 1H), 7.90 (d, J = 8.2 Hz, 2H), 7.33 (d, J = 8.2 Hz, 2H), 7.13 (d, J = 6.7) Hz, 1H), 7.03 (bs, 1H), 6.95 (ddd, J=7.6, 7.6, 1.6 Hz, 1H), 6.76 (dd, J=6.6, 1.6 Hz, 1H), 6.58 (ddd, J=7.4, 7.4, 1.7 Hz, 1H), 6.52 (s, 1H), 4.89 (bs, 2H), 3.95 (s, 2H). LRMS: 348.1 (calculated); 349.2 (exp.).
2－胺基－5－[4－(2－胺基－苯胺甲醯基)－苄基]－噻吩－3－甲酸醯胺(92)步驟2：4－(5－胺基－4－胺甲醯基－噻吩－2－基甲基)－苯甲酸甲酯(91) 依照與實例58步驟2(以2－氰基－乙醯胺置換丙二腈)所述相同之程序獲得呈黃色固體之標題化合物91(產率88%)。LRMS：304.1(計算值)；305.1(實驗值)。 2-Amino-5-[4-(2-amino-anilinomethyl)-benzyl]-thiophene-3-carboxylic acid decylamine (92) Step 2: 4-(5-Amino-4-amine Methylmercapto-thiophen-2-ylmethyl)-benzoic acid methyl ester (91) was obtained as a yellow solid according to the same procedure as in Example 58 Step 2 (M.sub. The title compound 91 (yield 88%). LRMS: 304.1 (calculated); 305.1 (experimental).
步驟3－4：2－胺基－5－[4－(2－胺基－苯胺甲醯基)－苄基]－噻吩－3－甲酸醯胺(92) 依照與實例1步驟4及5所述相同之程序自化合物91 開始獲得呈橙色固體之標題化合物92(產率54%)。1 H NMR：(DMSO)δ(ppm)：9.62(s,1H),7.90(d,J＝8.2 Hz,1H),7.32(d,J＝8.2 Hz,1H),7.14－7.12(m,3H),6.95(td,J＝7.6,1.6 Hz,1H),6.84(s,1H),6.76(dd,J＝7.8,1.2 Hz,1H),6.58(td,J＝7.4,1.2 Hz,1H),4.87(s,2H),3.93(s,2H)。LRMS：366.1(計算值)；367.4(實驗值)。 Step 3-4: 2-Amino-5-[4-(2-amino-anilinomethyl)-benzyl]-thiophene-3-carboxylic acid decylamine (92) according to steps 4 and 5 of Example 1. the same procedure described obtained starting from compound 91 as an orange solid of the title compound 92 (54% yield). 1 H NMR: (DMSO) δ (ppm): 9.62 (s, 1H), 7.90 (d, J = 8.2 Hz, 1H), 7.32 (d, J = 8.2 Hz, 1H), 7.14 - 7.12 (m, 3H) ), 6.95 (td, J = 7.6, 1.6 Hz, 1H), 6.84 (s, 1H), 6.76 (dd, J = 7.8, 1.2 Hz, 1H), 6.58 (td, J = 7.4, 1.2 Hz, 1H) , 4.87 (s, 2H), 3.93 (s, 2H). LRMS: 366.1 (calculated); 367.4 (exp.).
N－(2－胺基－苯基)－4－(4－氰基－噻吩－2－基甲基)－苯甲醯胺(94)步驟4：4－(4－氰基－噻吩－2－基甲基)－苯甲酸(93) 依照實例1步驟4(酯水解)及Tetrahedron Lett.；2001；42(32)；5367－5370所述之程序(脫胺化)自氰基化合物85開始獲得呈褐色固體之標題化合物93(產率76%)。LRMS：243.1(計算值)，244.3(實驗值)。 N-(2-Amino-phenyl)-4-(4-cyano-thiophen-2-ylmethyl)-benzamide (94) Step 4: 4-(4-Cyano-thiophene-2 -ylmethyl)-benzoic acid (93) according to the procedure described in Example 1 Step 4 (ester hydrolysis) and Tetrahedron Lett.; 2001; 42 (32); 5367-5370 (deamination) starting from cyano compound 85 The title compound 93 was obtained as a brown solid (yield: 76%). LRMS: 243.1 (calculated), 244.3 (exp.).
步驟5：N－(2－胺基－苯基)－4－(4－氰基－噻吩－2－基甲基)－苯甲醯胺(94) 依照與實例1步驟5所述相同之程存自化合物93閉始獲得呈橙色回體之標題化合物94(產率41%)。1 H NMR：(DMSO)δ(ppm)：9.61(bs,1H),8.38(d,J＝1.4 Hz,1H),7.92(d,J＝8.0 Hz,2H),7.40(d,J＝8.2 Hz,2H),7.33(d,J＝1.4 Hz,1H),7.13(d,J＝6.8 Hz,1H),6.95(td,J＝7.6,1.6 Hz,1H),6.75(dd,J＝8.0,1.4 Hz,1H),6.57(td,J＝7.4,1.2 Hz,1H),4.89(bs,2H),4.27(s,2H)。LRMS：333.1(計算值)；334.4(實驗值)。 Step 5: N-(2-Amino-phenyl)-4-(4-cyano-thiophen-2-ylmethyl)-benzimidamide (94) according to the same procedure as described in Step 5 of Example 1. The title compound 94 (yield 41%) was obtained as an orange residue from compound. 1 H NMR: (DMSO) δ (ppm): 9.61 (bs, 1H), 8.38 (d, J = 1.4 Hz, 1H), 7.92 (d, J = 8.0 Hz, 2H), 7.40 (d, J = 8.2 Hz, 2H), 7.33 (d, J = 1.4 Hz, 1H), 7.13 (d, J = 6.8 Hz, 1H), 6.95 (td, J = 7.6, 1.6 Hz, 1H), 6.75 (dd, J = 8.0 , 1.4 Hz, 1H), 6.57 (td, J = 7.4, 1.2 Hz, 1H), 4.89 (bs, 2H), 4.27 (s, 2H). LRMS: 333.1 (calculated); 334.4 (exp.).
4－(3－胺基－4－氰基－吡唑－1－基甲基)－N－(2－胺基－苯基)－苯甲醯胺(97)步驟1：4－(3－胺基－4－氰基－吡唑－1－基甲基)－苯甲酸甲酯(95) 向3－胺基－1H－吡唑－4－腈(1.211 g，11.21 mmol)之無水DMF(20 ml)溶液中加入K2 CO3 (5.414 g，39.24 mmol)。在室溫下攪拌該懸浮液5分鐘且將其以對溴甲基苯甲酸甲酯(2.568 g，11.21 mmol)處理，在100℃下攪拌2.5小時，冷卻且過濾之。蒸發濾液形成含油殘餘物，將該殘餘物溶解於Et2 －O－丙酮之混合物中且在－10℃下保存隔夜。形成結晶材料，將其以熱的EtOAc濕磨，再次在－10℃下保存隔夜且經過濾收集以形成標題化合物95(675 mg，產率24%)。LRMS：256.3(計算值)，257.3(實驗值)。 4-(3-Amino-4-cyano-pyrazol-1-ylmethyl)-N-(2-amino-phenyl)-benzamide (97) Step 1: 4-(3- Amino-4-cyano-pyrazol-1-ylmethyl)-benzoic acid methyl ester (95) to 3-amino-1H-pyrazole-4-carbonitrile (1.211 g, 11.21 mmol) in anhydrous DMF ( 20 ml) solution was added with K 2 CO 3 (5.414 g, 39.24 mmol). The suspension was stirred at room temperature for 5 minutes and treated with methyl p-bromomethylbenzoate (2.568 g, 11.21 mmol), stirred at 100 ° C for 2.5 h, cooled and filtered. The filtrate was evaporated formed oily residue, the residue was dissolved in a mixture of Et 2 -O- in acetone and stored overnight at -10 ℃. A crystalline material was formed which was triturated with hot EtOAc, EtOAc (EtOAc) EtOAc. LRMS: 256.3 (calculated), 257.3 (exp.).
步驟2：4－(3－胺基－4－氰基－吡唑－1－基甲基)－苯甲酸(96) 向溶於水(10 ml)及MeOH(20 ml)之混合物中之NaOH(343 mg，8.58 mmol)溶液中加入酯95(732 mg，2.86 mmol)。將反應混合物回流2分鐘且在周圍溫度下再攪拌2小時。減壓移除MeOH且以濃HCl(pH 3－4)酸化剩餘水溶液以形成沉澱物，過濾收集該沉澱物產生標題化合物(96 )(640 mg，產率92%)。LRMS：242.2(計算值)，241.1(實驗值)。 Step 2: NaOH in a mixture of 4-(3-amino-4-cyano-pyrazol-1-ylmethyl)-benzoic acid (96) in water (10 ml) and MeOH (20 ml) (95 mg, 8.58 mmol) was added ester 95 (732 mg, 2.86 mmol). The reaction mixture was refluxed for 2 minutes and stirred at ambient temperature for a further 2 hours. MeOH was removed under reduced pressure and with concentrated HCl (pH 3-4) the remaining aqueous solution was acidified to form a precipitate, the precipitate was collected by filtration to give the title compound (96) (640 mg, 92% yield). LRMS: 242.2 (calculated), 241.1 (exp.).
步驟3：4－(3－胺基－4－氰基－吡唑－1－基甲基)－N－(2－胺基－苯基)－苯甲醯胺(97) 依照與實例1步驟5所述相同之程序自化合物96開始獲得呈白色固體之標題化合物97。經急驟層析法(溶離劑MeOH－CH2 Cl2 (2：23))純化該粗產物隨後以CH2 Cl2 濕磨產生標題化合物97(產率58%)。1 H NMR：(400 MHz,DMSO－d6 ,δ(ppm)：9.61(s,1H),8.27(s,1H),7.92(d,J＝8.2,2H),7.33(d,J＝8.2,2H),7.13(d(dd),J＝7.8,1H),6.95(dd,J＝1.4 Hz,J＝7.8 Hz,1H),6.75(dd,J＝1.2 Hz,j＝7.8 Hz,1H),6.57(dd,J＝1.2 Hz,7.6 Hz,1H),5.60(s,2H),5.16(s,2H),4.89(s,2H)。LRMS：332.4(計算值)，333.4(實驗值)。 Step 3: 4-(3-Amino-4-cyano-pyrazol-1-ylmethyl)-N-(2-amino-phenyl)-benzamide (97) according to the procedure of Example 1. The title compound 97 was obtained as a white solid. By flash chromatography (eluent MeOH-CH 2 Cl 2 (2:23 )) The crude product was purified CH 2 Cl 2 followed by wet grinding to give the title compound 97 (58% yield). 1 H NMR: (400 MHz, DMSO-d 6 , δ (ppm): 9.61 (s, 1H), 8.27 (s, 1H), 7.92 (d, J = 8.2, 2H), 7.33 (d, J = 8.2 , 2H), 7.13 (d (dd), J = 7.8, 1H), 6.95 (dd, J = 1.4 Hz, J = 7.8 Hz, 1H), 6.75 (dd, J = 1.2 Hz, j = 7.8 Hz, 1H ), 6.57 (dd, J = 1.2 Hz, 7.6 Hz, 1H), 5.60 (s, 2H), 5.16 (s, 2H), 4.89 (s, 2H). LRMS: 332.4 (calculated), 333.4 (experimental value) ).
5－(2,4－二甲氧基－苄胺基)－苯幷呋喃－2－甲酸(2－胺基－苯基)－醯胺(102)步驟1－2：5－胺基－苯幷呋喃－2－甲酸甲酯(100) 依照J.Am.Chem.Soc.2000,122,(6382－6394)所述之程序自2－羥基－5－硝基－苯甲醛(98)開始經由中間體酯99獲得標題化合物100(產率74%)。 5-(2,4-Dimethoxy-benzylamino)-benzofuran-2-carboxylic acid (2-amino-phenyl)-guanamine (102) Step 1-2: 5-Amino-Benzene Methyl furfuran -2-carboxylate (100) was started from 2-hydroxy-5-nitro-benzaldehyde (98) according to the procedure described in J. Am. Chem. Soc. 2000, 122, (6382-6394) Intermediate ester 99 gave the title compound 100 (yield: 74%).
步驟3－4：5－(2,4－二甲氧基－苄胺基)－苯幷呋喃－2－甲酸(2－胺基－苯基)－醯胺(102) 依照與實例12步驟2(流程3)及實例1步驟4及5(流程1)相同之程序經由中間體100及101獲得呈橙色固體之標題化合物102(76 mg，41%)。1 H NMR：(DMSO)δ(ppm)：9.69(s,1H),7.44(s,1H),7.37(d,J＝8.8 Hz,1H),7.17－7.15(m,2H),6.96(td,J＝7.5,1.4 Hz,1H),6.84(dd,J＝9.0,2.3 Hz,1H),6.76(dd,J＝8.0,2.3 Hz,1H),6.67(d,J＝2.3 Hz,1H),6.60－6.56(m,2H),6.45(dd,J＝8.2,2.3 Hz,1H),5.94(t,J＝6.0 Hz,1H),4.92(s,2H),4.15(d,J＝5.7 Hz,1H),3.83(s,3H),3.73(s,3H)。LRMS：417.2(計算值)；418.5(獲得值)。 Step 3-4: 5-(2,4-Dimethoxy-benzylamino)-benzofuran-2-carboxylic acid (2-amino-phenyl)-decylamine (102) according to Step 2 of Example 12. (Scheme 3) The title compound 102 (76 mg, 41%) 1 H NMR: (DMSO) δ (ppm): 9.69 (s, 1H), 7.44 (s, 1H), 7.37 (d, J = 8.8 Hz, 1H), 7.17-7.15 (m, 2H), 6.96 (td) , J=7.5, 1.4 Hz, 1H), 6.84 (dd, J=9.0, 2.3 Hz, 1H), 6.76 (dd, J=8.0, 2.3 Hz, 1H), 6.67 (d, J=2.3 Hz, 1H) , 6.60-6.56 (m, 2H), 6.45 (dd, J = 8.2, 2.3 Hz, 1H), 5.94 (t, J = 6.0 Hz, 1H), 4.92 (s, 2H), 4.15 (d, J = 5.7 Hz, 1H), 3.83 (s, 3H), 3.73 (s, 3H). LRMS: 417.2 (calculated); 418.5 (obtained).
N－(2－胺基－苯基)－4－[5－(3－甲氧基－苄基)－[1,2,4]噁二唑－3－基甲基]－苯甲醯胺(105)步驟1：4－[5－(3－甲氧基－苄基)－[1,2,4]噁二唑－3－基甲基]－苯甲酸(104) 向4－(N－羥基脲基甲基)－苯甲酸(103)(描述於專利申請案WO 03/024448中)(464 mg，2.40 mmol)之無水吡啶(10 ml)懸浮液中加入(3－甲氧基－苯基)－乙醯氯(418 mg，2.27 mmol)且將反應混合物回流3小時，冷卻，將其以水(10 ml)中止且蒸發形成固體殘餘物。將該材料再溶解於CH2 Cl2 中，以木炭脫色且以CH2 Cl2 －MeOH(19：1)、CH2 Cl2 －丙酮(19：1，接著9：1)及丙酮－己烷(3：2)為溶離劑由急驟層析法3次純化該材料產生標題化合物104(96 mg，13%)。LRMS：324.3(計算值)，323.3[M－H]－ (實驗值)。 N-(2-Amino-phenyl)-4-[5-(3-methoxy-benzyl)-[1,2,4]oxadiazol-3-ylmethyl]-benzamide (105) Step 1: 4-[5-(3-Methoxy-benzyl)-[1,2,4]oxadiazol-3-ylmethyl]-benzoic acid (104) to 4-(N) 3-Hydroxyureidomethyl)-benzoic acid (103) (described in patent application WO 03/024448) (464 mg, 2.40 mmol) in anhydrous pyridine (10 ml) was added (3-methoxy- Phenyl)-ethyl hydrazine chloride (418 mg, 2.27 mmol) and the reaction mixture was refluxed for 3 hrs, cooled and evaporated with water (10 ml). The material was redissolved in CH 2 Cl 2 and decolorized with charcoal with CH 2 Cl 2 -MeOH (19:1), CH 2 Cl 2 -acetone (19:1, followed by 9:1) and acetone-hexane (3:2) Purification of the material by flash chromatography eluting to afford title compound 104 (96 mg, 13%). LRMS: 324.3 (calculated), 323.3 [M-H] - (exp.).
N－(2－胺基－苯基)－4－[5－(3－甲氧基－苄基)－[1,2,4]噁二唑－3－基甲基]－苯甲醯胺(105) 依照與實例1步驟5(流程1)相同之程序獲得呈白色固體之標題化合物105。以MeOH－CH2 Cl2 (1：19)接著EtOAc－CH2 Cl2 (1：2)為溶離劑由急驟層析法兩次純化該粗產物得到產率為41%之標題化合物。LRMS：414.5(計算值)，415.4[MH]＋ (實驗值)。1 H NMR：(400 MHz,DMSO－d6 ,δ(ppm)：9.61(s,1H),7.91(d,J＝8.22,2H),7.41(d,J＝8.22,2H),7.25(t,J＝7.83,1H),7.14(d,J＝6.65,1H),6.96(m,1H),6.89－6.84(m,3H),6.76(dd,J＝8.02,1.37,1H),6.58(dt,J＝7.63,1.30,1H),4.89(s,2H),4.29(s,2H),4.16(s,2H),3.73(s,3H)。 N-(2-Amino-phenyl)-4-[5-(3-methoxy-benzyl)-[1,2,4]oxadiazol-3-ylmethyl]-benzamide (105) The title compound 105 was obtained as a white solid. In MeOH-CH 2 Cl 2 (1:19 ) followed by EtOAc-CH 2 Cl 2 (1 : 2) as eluent purified twice by flash chromatography of the crude product was obtained in a yield of 41% of the title compound. LRMS: 414.5 (calculated), 415.4 [MH] + (yield). 1 H NMR: (400 MHz, DMSO-d 6 , δ (ppm): 9.61 (s, 1H), 7.91 (d, J = 8.22, 2H), 7.41 (d, J = 8.22, 2H), 7.25 (t , J = 7.83, 1H), 7.14 (d, J = 6.65, 1H), 6.96 (m, 1H), 6.89 - 6.84 (m, 3H), 6.76 (dd, J = 8.02, 1.37, 1H), 6.58 ( Dt, J = 7.63, 1.30, 1H), 4.89 (s, 2H), 4.29 (s, 2H), 4.16 (s, 2H), 3.73 (s, 3H).
5－(4－酮基－4H－噻吩幷[3,2－d]嘧啶－3－基甲基)－苯幷呋喃－2－甲酸(2－胺基－苯基)－醯胺(111) 依照專利申請案WO 03/024448所述之程序自化合物106開始[與3H－噻吩幷[3,2－d]嘧啶－4－酮(108 )反應]經由中間體109獲得標題化合物111。1 H NMR：(DMSO)δ(ppm)：9.87(s,1H)；8.72(s,1H)；8.20(d,J＝5.5 Hz,1H)；7.81(s,1H)；7.68(m,2H)；7.54(dd,J＝8.6,1.6 Hz,1H),7.42(d,J＝5.5 Hz,1H)；7.17(d,J＝6.6,1H)；6.98(dt,J＝7.8,1.6 Hz,1H)；6.77(dd,J＝8.2,1.6 Hz,1H),6.59(dt,J＝7.4,1.6 Hz,1H)；5.35(s,2H)；4.96(s,2H)。LRMS：416.1(計算值)，417.4(獲得值)。 5-(4-keto-4H-thiophene[3,2-d]pyrimidin-3-ylmethyl)-benzofuran-2-carboxylic acid (2-amino-phenyl)-decylamine (111) Starting from compound 106 [reacting with 3H-thiophene[3,2-d]pyrimidin-4-one ( 108 )] according to the procedure described in the patent application WO 03/024448, the title compound 111 is obtained via intermediate 109. 1 H NMR: (DMSO) δ (ppm): 9.87 (s, 1H); 8.72 (s, 1H); 8.20 (d, J = 5.5 Hz, 1H); 7.81 (s, 1H); 7.68 (m, 2H) ); 7.54 (dd, J = 8.6, 1.6 Hz, 1H), 7.42 (d, J = 5.5 Hz, 1H); 7.17 (d, J = 6.6, 1H); 6.98 (dt, J = 7.8, 1.6 Hz, 1H); 6.77 (dd, J = 8.2, 1.6 Hz, 1H), 6.59 (dt, J = 7.4, 1.6 Hz, 1H); 5.35 (s, 2H); 4.96 (s, 2H). LRMS: 416.1 (calculated), 417.4 (obtained).
5－(4－酮基－4H－噻吩幷[3,2－d]嘧啶－3－基甲基)－苯幷[b]噻吩－2－甲酸(2－胺基－苯基)－醯胺(112) 依照專利申請案WO 03/024448所述之程序自化合物107開始[與3H－噻吩幷[3,2－d]嘧啶－4－酮(108)反應]經由中間體110獲得標題化合物112。1 H NMR：(DMSO)δ(ppm)：9.89(s,1H),8.74(s,1H),8.29(s,1H),8.21(d,J＝5.28,1H),8.03(d,J＝8.22,1H),7.98(s,1H),7.51(dd,J＝8.51,1.67,1H),7.43(d,J＝5.28,1H),7.17(dd,J＝7.73,1.27,1H),6.99(m,1H),6.79(dd,J＝8.22,1.37,1H),6.60(dt,J＝7.43,1.37,1H),5.38(s,2H),5.00(s,2H)。LRMS：432.1(計算值)，433.3(獲得值)。 5-(4-keto-4H-thiophene[3,2-d]pyrimidin-3-ylmethyl)-benzoquinone[b]thiophene-2-carboxylic acid (2-amino-phenyl)-decylamine (112) Starting from compound 107 [reacting with 3H-thiophene[3,2-d]pyrimidin-4-one (108)] according to the procedure described in the patent application WO 03/024448, the title compound 112 is obtained via intermediate 110 . 1 H NMR: (DMSO) δ (ppm): 9.89 (s, 1H), 8.74 (s, 1H), 8.29 (s, 1H), 8.21 (d, J = 5.28, 1H), 8.03 (d, J = 8.22,1H), 7.98(s,1H), 7.51 (dd, J=8.51, 1.67,1H), 7.43 (d, J=5.28,1H), 7.17 (dd, J=7.73, 1.27,1H), 6.99 (m, 1H), 6.79 (dd, J = 8.22, 1.37, 1H), 6.60 (dt, J = 7.43, 1.37, 1H), 5.38 (s, 2H), 5.00 (s, 2H). LRMS: 432.1 (calculated), 433.3 (obtained).
步驟5：2－(4－三氟甲氧基－苄胺基)－苯幷噻唑－6－甲酸(2－胺基－苯基)－醯胺(117)步驟1：2－胺基－苯幷噻唑－6－甲酸甲酯(113) 依照J.Med.Chem.1997；40(105－111)所述之程序自4－胺基－苯甲酸甲酯開始獲得標題化合物。 Step 5: 2-(4-Trifluoromethoxy-benzylamino)-benzothiazole-6-carboxylic acid (2-amino-phenyl)-decylamine (117) Step 1: 2-Amino-Benzene The title compound was obtained starting from 4-amino-benzoic acid methyl ester according to the procedure described in J. Med. Chem. 1997; 40 (105-111).
步驟2：2－胺基－苯幷噻唑－6－甲酸(114) 依照實例1步驟4所述之程序獲得標題化合物114(產率97%)。1 H－NMR(DMSO)δ：12.58(s,1H),8.23(d,J＝1.8 Hz,1H)；7.85(s,2H)；7.78(dd,J＝8.4,1.8 Hz,1H)；7.33(d,J＝7.8 Hz,1H)。 Step 2: 2-Amino-benzothiazole-6-carboxylic acid (114) The title compound (yield: 97%) was obtained according to the procedure 1 H-NMR (DMSO) δ: 12.58 (s, 1H), 8.23 (d, J = 1.8 Hz, 1H); 7.85 (s, 2H); 7.78 (dd, J = 8.4, 1.8 Hz, 1H); (d, J = 7.8 Hz, 1H).
步驟3：{2－[(2－胺基－苯幷噻唑－6－羰基)－胺基]－苯基}－胺基甲酸第三丁酯(115) 在DMF中使酸114(1.80 g，9.27 mmol)與(2－胺基－苯基)－胺基甲酸第三丁酯(2.31 g，11.1 mmol)及BOP(4.91 g，11.1 mmol)組合。向該溶液中加入Et3 N(5.16 ml，37.1 mmol)且將混合物在室溫下氮氣下攪拌隔夜，將其在真空中濃縮且由急驟柱層析法(30%己烷/EtOAc)加以純化。為進一步純化該產物，使混合物在EtOAc與水之間分溶，分離有機層，使其經MgSO4 乾燥及蒸發產生標題化合物115(1.84 g，52%)。1 H－NMR(DMSO)δ：9.72(s,1H),8.66(m,1H)；8.22(d,J＝1.8 Hz,1H)；7.80(m,3H)；7.50(m,2H)；7.37(m,1H)；7.14(m,2H)；1.44(s,9H)。 Step 3: {2-[(2-Amino-benzothiazole-6-carbonyl)-amino]-phenyl}-carbamic acid tert-butyl ester (115) mp 14 (1.80 g, 9.27 mmol) in combination with (2-amino-phenyl)-carbamic acid tert-butyl ester (2.31 g, 11.1 mmol) and BOP (4.91 g, 11.1 mmol). To this solution was added Et 3 N (5.16 ml, 37.1 mmol) and the mixture was stirred overnight under nitrogen at room temperature, it was concentrated in vacuo and purified by flash column chromatography (30% hexanes / EtOAc) . For further purification of the product, the mixture was partitioned between EtOAc and water, the organic layer was separated, to produce the title compound 115 (1.84 g, 52%) and dried over MgSO 4 and evaporated. 1 H-NMR (DMSO) δ: 9.72 (s, 1H), 8.66 (m, 1H); 8.22 (d, J = 1.8 Hz, 1H); 7.80 (m, 3H); 7.50 (m, 2H); (m, 1H); 7.14 (m, 2H); 1.44 (s, 9H).
步驟4：2－(4－三氟甲氧基－苯基)－苯幷噻唑－6－甲酸(2－胺基－苯基)－醯胺(116) 向115(300 mg，0.78 mmol)、4－(三氟甲氧基)苯甲醛(123μl，0.86 mmol)及氯化二丁基錫(24 mg，0.08 mmol)之THF溶液中加入苯矽烷(106μl，0.86 mmol)。在室溫氮氣下將該混合物攪拌隔夜，再加入甲醛及苯矽烷且繼續攪拌至無原料存在。自混合物蒸除THF且由急驟柱層析法(EtOAc/己烷30/70，接著50/50)純化該殘餘物產生標題化合物116(314 mg,72%)。1 H－NMR(DMSO)δ：9.77(s,1H),8.89(t,J＝5.7 Hz,1H)；8.69(s,1H)；8.29(d,J＝1.8 Hz,1H)；7.84(dd,J＝8.4,1.8 Hz,1H)；7.50(m,5H)；7.37(d,J＝7.8 Hz,2H)；7.17(m,2H)；4.69(d,J＝5.7 Hz,2H)；1.47(s,9H)。 Step 4: 2-(4-Trifluoromethoxy-phenyl)-benzothiazole-6-carboxylic acid (2-amino-phenyl)-guanamine (116) to 115 (300 mg, 0.78 mmol), To a solution of 4-(trifluoromethoxy)benzaldehyde (123 μl, 0.86 mmol) and dibutyltin chloride (24 mg, 0.08 mmol) in THF was added phenyl decane (106 μl, 0.86 mmol). The mixture was stirred overnight under nitrogen at room temperature, then formaldehyde and benzoquinane were added and stirring was continued until no material was present. The residue was purified by flash chromatography eluting elut elut elut elut elut elut elut 1 H-NMR (DMSO) δ: 9.77 (s, 1H), 8.89 (t, J = 5.7 Hz, 1H); 8.69 (s, 1H); 8.29 (d, J = 1.8 Hz, 1H); , J = 8.4, 1.8 Hz, 1H); 7.50 (m, 5H); 7.37 (d, J = 7.8 Hz, 2H); 7.17 (m, 2H); 4.69 (d, J = 5.7 Hz, 2H); 1.47 (s, 9H).
步驟5：2－(4－三氟甲氧基－苄胺基)－苯幷噻唑－6－甲酸(2－胺基－苯基)－醯胺(117) 向116(306 mg，0.55 mmol)之DCM溶液中加入TFA(2.0 ml)。在室溫下攪拌該混合物4小時且濃縮之。將殘餘物溶解於EtOAc中，將其以NaHCO3 洗滌，經MgSO4 乾燥且再次濃縮之。由急驟柱層析法(EtOAc中之30%己烷)純化該殘餘物產生呈黃色固體之標題化合物117(252 mg，100%)。1 H－NMR(DMSO)δ：9.56(s,1H),8.83(t,J＝5.8 Hz,1H),8.30(d,J＝1.8 Hz,1H)；7.85(dd,J＝8.4,1.6 Hz,1H)；7.49(d,J＝8.4 Hz,2H)；7.43(d,J＝8.4 Hz,1H)；7.34(d,J＝8.4 Hz,2H)；7.15(d,J＝7.6 Hz.1H)；6.94(brt,J＝7.8 Hz,1H)；6.77(d,J＝7.8 Hz,1H)；6.59(t,J＝7.5 Hz,1H)；4.66(d,J＝5.7 Hz,2H)。LRMS：458.1(計算值)，459.2(獲得值)。 Step 5: 2-(4-Trifluoromethoxy-benzylamino)-benzothiazole-6-carboxylic acid (2-amino-phenyl)-guanamine (117) to 116 (306 mg, 0.55 mmol) TFA (2.0 ml) was added to the DCM solution. The mixture was stirred at room temperature for 4 hours and concentrated. The residue was dissolved in EtOAc, dried which was washed with NaHCO 3 over MgSO 4 and concentrated once again. The residue was purified by flash chromatography eluting elut elut elut elut elut elut elut elut 1 H-NMR (DMSO) δ: 9.56 (s, 1H), 8.83 (t, J = 5.8 Hz, 1H), 8.30 (d, J = 1.8 Hz, 1H); 7.85 (dd, J = 8.4, 1.6 Hz , 1H); 7.49 (d, J = 8.4 Hz, 2H); 7.43 (d, J = 8.4 Hz, 1H); 7.34 (d, J = 8.4 Hz, 2H); 7.15 (d, J = 7.6 Hz. 1H) 6.94 (brt, J = 7.8 Hz, 1H); 6.77 (d, J = 7.8 Hz, 1H); 6.59 (t, J = 7.5 Hz, 1H); 4.66 (d, J = 5.7 Hz, 2H). LRMS: 458.1 (calculated), 459.2 (obtained).
6－(3,4－二甲氧基－苄胺基)－苯幷噁唑－2－甲酸(2－胺基－苯基)－醯胺(120)步驟1：5－胺基－苯幷噁唑－2－甲酸甲酯(118) 依照J.Am.Chem.Soc.2000； 122(6382－6394)所述之程序自苯幷噁唑－2－甲酸甲酯開始獲得標題化合物118 。 6-(3,4-Dimethoxy-benzylamino)-benzoxazole-2-carboxylic acid (2-amino-phenyl)-guanamine (120) Step 1: 5-Amino-benzoquinone The title compound 118 was obtained starting from methyl benzoxazole-2-carboxylate according to the procedure described in J. Am. Chem. Soc. 2000; 122 (6382-6394).
步驟2：5－(3,4－二甲氧基－苄胺基)－苯幷噁唑－2－甲酸甲酯(119) 依照與實例68步驟4(流程28)所述相同之程序獲得固體標題化合物119(產率90%)。LRMS：342.1(計算值)；343.4(實驗值)。 Step 2: 5-(3,4-Dimethoxy-benzylamino)-benzoxazole-2-carboxylic acid methyl ester (119) was obtained according to the same procedure as described in Example 68 Step 4 (Schem. 28). The title compound 119 (yield 90%). LRMS: 342.1 (calculated); 343.4 (exp.).
步驟4－5：6－(3,4－二甲氧基－苄胺基)－苯幷噁唑－2－甲酸(2－胺基－苯基)－醯胺(120) 依照實例1步驟5(流程1)所述之程序獲得標題化合物120(產率31%)。1 H NMR：(DMSO)δ(ppm)：10.06(s,1H),7.56(d,J＝8.80,1H),7.22(dd,J＝7.83,1.37,1H),7.02(d,J＝1.57,1H),6.98(m,1H),6.92(d,J＝1.76,1H),6.88(d,J＝1.96,1H),6.86(d,J＝2.35,1H),6.82(d,J＝1.96,1H),6.78(dd,J＝7.93,1.27,1H),6.60(m,1H),4.99(brs,2H),4.28(d,J＝5.48,2H),3.76(s,3H),3.73(s,3H)。LRMS：418(計算值)，419.5(獲得值)。 Step 4-5: 6-(3,4-Dimethoxy-benzylamino)-benzoxazole-2-carboxylic acid (2-amino-phenyl)-decylamine (120) according to Example 1 Step 5 The procedure described in (Scheme 1) gave the title compound 120 (yield 31%). 1 H NMR: (DMSO) δ (ppm): 10.06 (s, 1H), 7.56 (d, J = 8.80, 1H), 7.22 (dd, J = 7.83, 1.37, 1H), 7.02 (d, J = 1.57) , 1H), 6.98 (m, 1H), 6.92 (d, J = 1.76, 1H), 6.88 (d, J = 1.96, 1H), 6.86 (d, J = 2.35, 1H), 6.82 (d, J = 1.96,1H), 6.78 (dd, J=7.93, 1.27,1H), 6.60 (m, 1H), 4.99 (brs, 2H), 4.28 (d, J = 5.48, 2H), 3.76 (s, 3H), 3.73 (s, 3H). LRMS: 418 (calculated), 419.5 (obtained).
1－(3,4,5－三甲氧基－苄基)－2,3－二氫－1H－吲哚－5－甲酸(2－胺基－苯基)－醯胺(122) 依照實例68步驟4(流程28)(用於製備中間體121)所述之程序及實例1步驟4及5(流程1)所述之程序獲得標題化合物122(產率33%)。1 H－NMR(DMSO)δ：9.29(s,1H),7.71(dd,J＝8.22,1.77,1H),7.66(brm,1H),7.12(dd,J＝7.93,1.47,1H),6.93－6.89(m,3H),6.84(dd,J＝8.22,1.96,1H),6.75(dd,J＝8.02,1.37,1H),6.65(d,J＝8.41,1H),6.57(dt,J＝7.53,1.30,1H),4.82(s,2H),3.73(s,6H),3.41(t,J＝8.51,2H),2.98(t,J＝8.51,2H)。LRMS：435.2(計算值)；436.5(獲得值)。 1- (3,4,5-trimethoxy - benzyl) -2,3-dihydro -1H- indole-5-carboxylic acid (2-amino - phenyl) - Amides (122) in accordance with Example 68 The procedure described in Step 4 (Scheme 28) (for the preparation of Intermediate 121) and the procedure of Example 1 Steps 4 and 5 (Scheme 1) afforded the title compound 122 (yield 33%). 1 H-NMR (DMSO) δ: 9.29 (s, 1H), 7.71 (dd, J = 8.22, 1.77, 1H), 7.66 (brm, 1H), 7.12 (dd, J = 7.93, 1.47, 1H), 6.93 -6.89 (m, 3H), 6.84 (dd, J = 8.22, 1.96, 1H), 6.75 (dd, J = 8.02, 1.37, 1H), 6.65 (d, J = 8.41, 1H), 6.57 (dt, J) = 7.53, 1.30, 1H), 4.82 (s, 2H), 3.73 (s, 6H), 3.41 (t, J = 8.51, 2H), 2.98 (t, J = 8.51, 2H). LRMS: 435.2 (calculated); 436.5 (obtained).
[6－(2－胺基－苯胺甲醯基)－1－甲基－1H－吲哚－3－基甲基]－(4－嗎啉－4－基－苯基)－胺基甲酸第三丁酯(126)步驟1：3－[(4－嗎啉－4－基－苯胺基)－甲基]－1H－吲哚－6－甲酸甲酯(123) 向3－甲醯基－1H－吲哚－6－甲酸甲酯(500 mg，2.46 mmol)、4－嗎啉幷苯胺(482.3 mg，2.71 mmol)及氯化二丁基錫(76 mg，0.25 mmol)之THF溶液中加入苯矽烷(334μl，2.71 mmol)。在室溫下氮氣下將混合物攪拌隔夜，自混合物蒸除THF且由急驟層析法(己烷/EtOAc，20/80)純化殘餘物產生標題化合物123(881 mg，98%)。1 H－NMR(DMSO)δ：8.00(d,J＝1.0 Hz,1H),7.70(d,J＝8.2 Hz,1H)；7.58(dd,J＝8.4,1.6 Hz,1H)；7.52(d,J＝2.0 Hz,1H)；6.70(d,J＝9.0 Hz,2H)；6.59(d,J＝9.0 Hz,2H)；5.48(t,J＝5.8 Hz,1H)；4.31(d,J＝5.7 Hz,2H)；3.83(s,3H)；3.68(t,J＝4.7 Hz,4H)；2.86(t,J＝4.7 Hz,4H)。 [6-(2-Amino-anilinemethanyl)-1-methyl-1H-indol-3-ylmethyl]-(4-morpholin-4-yl-phenyl)-carbamic acid Tributyl ester (126) Step 1: 3-[(4-morpholin-4-yl-anilino)-methyl]-1H-indole-6-carboxylic acid methyl ester (123) to 3 -methylindenyl- Addition of phenyl decane to 1H-indole-6-methyl formate (500 mg, 2.46 mmol), 4-morpholinium anilide (482.3 mg, 2.71 mmol) and dibutyltin chloride (76 mg, 0.25 mmol) in THF (334 μl, 2.71 mmol). The mixture was stirred at rt EtOAc (EtOAc)EtOAc. 1 H-NMR (DMSO) δ: 8.00 (d, J = 1.0 Hz, 1H), 7.70 (d, J = 8.2 Hz, 1H); 7.58 (dd, J = 8.4, 1.6 Hz, 1H); , J = 2.0 Hz, 1H); 6.70 (d, J = 9.0 Hz, 2H); 6.59 (d, J = 9.0 Hz, 2H); 5.48 (t, J = 5.8 Hz, 1H); 4.31 (d, J = 5.7 Hz, 2H); 3.83 (s, 3H); 3.68 (t, J = 4.7 Hz, 4H); 2.86 (t, J = 4.7 Hz, 4H).
步驟2：3－{[第三丁氧基羰基－(4－嗎啉－4－基－苯基)－胺基]－甲基}－1H－吲哚－6－甲酸甲酯(124) 向123(689 mg，1.89 mmol)之THF(100 ml)溶液中逐滴加入Et3 N(289μl，2.08 mmol)。緩緩加入(BOC)2 O且在室溫下氮氣下將混合物攪拌隔夜，蒸除THF且使殘餘物在水與CH2 Cl2 之間分溶。分離有機層，使其經MgSO4 乾燥，蒸發以形成另一殘餘物，由急驟柱層析法(EtOAc/己烷，7：3)純化該殘餘物產生標題化合物124(692 mg，79%)。1 H－NMR(CDCl3 )δ：8.24(m,1H),8.09(m,1H)；7.76(dd,J＝8.2,1.4 Hz,1H)；7.55(d,J＝8.8 Hz,1H)；7.13(d,J＝2.5 Hz,1H)；6.93(m,3H)；4.97(s,2H)；3.94(s,3H)；3.89(m,4H)；3.16(m,4H)；1.47(s,9H)。 Step 2: 3-{[Tertibutoxycarbonyl-(4-morpholin-4-yl-phenyl)-amino]-methyl}-1H-indole-6-carboxylic acid methyl ester (124) 123 (689 mg, 1.89 mmol) of THF (100 ml) was added dropwise Et 3 N (289μl, 2.08 mmol ). (BOC) 2 O was slowly added and the mixture was stirred overnight under nitrogen, THF was evaporated and residue was partitioned between water and CH 2 Cl 2 . The organic layer was separated, dried over MgSO 4, evaporated to form another residue was purified by flash column chromatography (EtOAc / hexane, 7: 3) to furnish the title compound was 124 (692 mg, 79%) . 1 H-NMR (CDCl 3 ) δ: 8.24 (m, 1H), 8.09 (m, 1H); 7.76 (dd, J = 8.2, 1.4 Hz, 1H); 7.55 (d, J = 8.8 Hz, 1H); 7.13 (d, J = 2.5 Hz, 1H); 6.93 (m, 3H); 4.97 (s, 2H); 3.94 (s, 3H); 3.89 (m, 4H); 3.16 (m, 4H); 1.47 (s) , 9H).
步驟3：3－{[第三丁氧基羰基－(4－嗎啉－4－基－苯基)－胺基]－甲基}－1－甲基－1H－吲哚－6－甲酸甲酯(125) 向酯124(473 mg，1.02 mmol)之DMF(15 ml)溶液中加入60%NaH(45 mg，1.12 mmol)。在室溫下氮氣下攪拌該溶液一小時，冷卻至0℃，將其以MeI(170μl，1.12 mmol)處理，溫至室溫並在氮氣下攪拌隔夜。使混合物在水與AcOEt之間分溶，收集有機層、使其經MgSO4 乾燥且在真空中濃縮以產生454 mg(93%)。1 H－NMR(DMSO)δ：7.99(m,1H),7.56(dd,J＝1.4,8.2 Hz,1H)；7.47(m,1H)；7.27(s,1H)；6.86(d,J＝8.8 Hz,2H)；6.75(d,J＝9.0 Hz,2H)；4.86(s,2H)；3.83(s,3H)；3.76(s,3H)；3.67(t,J＝4.8 Hz,4H)；3.01(t,J＝4.8 Hz,4H)；1.37(s,9H)。 Step 3: 3-{[Tertibutoxycarbonyl-(4-morpholin-4-yl-phenyl)-amino]-methyl}-1-methyl-1H-indole-6-carboxylic acid To a solution of the ester 124 (473 mg, 1.02 mmol) in DMF (15 ml), 60% NaH (45 mg, 1. The solution was stirred at room temperature under nitrogen for one hour, cooled to 0.degree. C., EtOAc (EtOAc, EtOAc) The mixture was partitioned between water and AcOEt, the organic layer was collected, it was dried over MgSO 4 and concentrated in vacuo to yield 454 mg (93%). 1 H-NMR (DMSO) δ : 7.99 (m, 1H), 7.56 (dd, J = 1.4,8.2 Hz, 1H); 7.47 (m, 1H); 7.27 (s, 1H); 6.86 (d, J = 8.8 Hz, 2H); 6.75 (d, J = 9.0 Hz, 2H); 4.86 (s, 2H); 3.83 (s, 3H); 3.76 (s, 3H); 3.67 (t, J = 4.8 Hz, 4H) ;3.01 (t, J = 4.8 Hz, 4H); 1.37 (s, 9H).
步驟4－5：[6－(2－胺基－苯胺甲醯基)－1－甲基－1H－吲哚－3－基甲基]－(4－嗎啉－4－基－苯基)－胺基甲酸第三丁酯(126) 依照實例1步驟4及5(流程1)描述之程序產生固體標題化合物126(134 mg，33%)。1 H－NMR(DMSO)δ：9.59(s,1H)；8.06(s,1H)；7.61(dd,J＝1.6,8.4 Hz,1H)；7.48(m,1H)；7.21(s,1H)；7.15(dd,J＝7.8,1.4 Hz,1H)；6.94(dt,J＝7.8,1.6 Hz,1H)；6.86(m,2H)；6.76(m,3H)；6.58(dt,J＝7.4,1.4 Hz,1H)；4.88(s,2H)；4.87(s,2H)；3.76(s,3H)；3.68(t,J＝4.8 Hz,4H)；3.02(t,J＝4.8 Hz,4H)；1.39(s,9H)。LRMS：556.2(計算值)；557.5(實驗值)。 Step 4-5: [6-(2-Amino-anilinemethanyl)-1-methyl-1H-indol-3-ylmethyl]-(4-morpholin-4-yl-phenyl) The title compound 126 (134 mg, 33%) was obtained according to the procedure described in Steps 4 and 5 (Scheme 1). 1 H-NMR (DMSO) δ: 9.59 (s, 1H); 8.06 (s, 1H); 7.61 (dd, J = 1.6, 8.4 Hz, 1H); 7.48 (m, 1H); 7.21 (s, 1H) ; 7.15 (dd, J = 7.8, 1.4 Hz, 1H); 6.94 (dt, J = 7.8, 1.6 Hz, 1H); 6.86 (m, 2H); 6.76 (m, 3H); 6.58 (dt, J = 7.4) , 1.4 Hz, 1H); 4.88 (s, 2H); 4.87 (s, 2H); 3.76 (s, 3H); 3.68 (t, J = 4.8 Hz, 4H); 3.02 (t, J = 4.8 Hz, 4H) ); 1.39 (s, 9H). LRMS: 556.2 (calculated); 557.5 (exp.).
N－(2－胺基－苯基)－4－[(6－羥基－苯幷噻唑－2－基胺基)－甲基]－苯甲醯胺(134)步驟1：(2－{4－[(6－羥基－苯幷噻唑－2－基胺基)－甲基]－苯甲醯胺基}－苯基)－胺基甲酸第三丁酯(129)： 依照與流程3步驟2(實例12)對還原胺化反應所述相同之程序自胺基噻唑127及醛128(專利申請案WO 03/024448中所述)獲得標題化合物129(產率96%)。1 H NMR：(丙酮－d6 )δ(ppm)：9.60(s,1H),8.25(bs,1H),7.99(d,J＝8.2 Hz,2H),7.69(d,J＝7.4 Hz,1H),7.61－7.58(m,3H),7.39(bs,1H),7.27(d,J＝8.6 Hz,1H),7.19(五重峰d,J＝7.4,2.0 Hz,2H),7.12(d,J＝2.3 Hz,1H),6.79(dd,J＝8.6,2.7 Hz,1H),4.78(s,2H),1.48(s,9H)。m/z：491.5(MH＋ )。 N-(2-Amino-phenyl)-4-[(6-hydroxy-benzothiazol-2-ylamino)-methyl]-benzamide (134) Step 1: (2-{4 -[(6-Hydroxy-benzothiazol-2-ylamino)-methyl]-benzylaminoamido}-phenyl)-carbamic acid tert-butyl ester (129): according to step 2 of Scheme 3 (Example 12) The same procedure as described for the reductive amination reaction. The title compound 129 (yield: 96%) was obtained from the amine thiazole 127 and aldehyde 128 (as described in the patent application WO 03/024448). 1 H NMR: (acetone-d 6 ) δ (ppm): 9.60 (s, 1H), 8.25 (bs, 1H), 7.99 (d, J = 8.2 Hz, 2H), 7.69 (d, J = 7.4 Hz, 1H), 7.61-7.58 (m, 3H), 7.39 (bs, 1H), 7.27 (d, J = 8.6 Hz, 1H), 7.19 (five-fold d, J = 7.4, 2.0 Hz, 2H), 7.12 ( d, J = 2.3 Hz, 1H), 6.79 (dd, J = 8.6, 2.7 Hz, 1H), 4.78 (s, 2H), 1.48 (s, 9H). m/z: 491.5 (MH + ).
步驟2：N－(2－胺基－苯基)－4－[(6－羥基－苯幷噻唑－2－基胺基)－甲基]－苯甲醯胺(134) 依照流程28步驟5(實例68)對Boc***所述之程序自化合物129開始獲得標題化合物134(產率53%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.58(s,1H),9.12(s,1H),8.25(t,J＝6.3 Hz,1H),7.91(d,J＝7.8 Hz,2H),7.45(d,J＝8.2 Hz,2H),7.15(d,J＝8.6 Hz,1H)7.12(s,1H),7.02(d,J＝2.7 Hz,1H),6.94(t,J＝6.7 Hz,1H),6.75(dd,J＝8.2,1.2 Hz,1H),6.63(dd,J＝8.6,2.3 Hz,1H),6.56(t,J＝7.8 Hz,1H),4.87(s,2H),4.59(d,J＝5.5 Hz,2H)。m/z：391.2(MH＋ ) Step 2: N-(2-Amino-phenyl)-4-[(6-hydroxy-benzothiazol-2-ylamino)-methyl]-benzamide (134) according to Step 28 of Scheme 28. (Example 68) Procedure for Boc cleavage The title compound 134 (yield: 53%) was obtained from compound 129. 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.58 (s, 1H), 9.12 (s, 1H), 8.25 (t, J = 6.3 Hz, 1H), 7.91 (d, J = 7.8 Hz, 2H), 7.45 (d, J = 8.2 Hz, 2H), 7.15 (d, J = 8.6 Hz, 1H) 7.12 (s, 1H), 7.02 (d, J = 2.7 Hz, 1H), 6.94 (t, J = 6.7 Hz, 1H), 6.75 (dd, J = 8.2, 1.2 Hz, 1H), 6.63 (dd, J = 8.6, 2.3 Hz, 1H), 6.56 (t, J = 7.8 Hz, 1H), 4.87 (s , 2H), 4.59 (d, J = 5.5 Hz, 2H). m/z: 391.2 (MH + )
N－(2－胺基－苯基)－4－{[6－(2－氯－乙氧基)－苯幷噻唑－2－基胺基]－甲基}－苯甲醯胺(135)步驟1：[2－(4－([6－(2－氯－乙氧基)－苯幷噻唑－2－基胺基]－甲基)－苯甲醯胺基)－苯基]－胺基甲酸第三丁酯(130) 依照J.Med.Chem.,2002,45(6),1300－1312所述之程序用化合物129作為原料獲得標題化合物130(產率43%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.59(bs,1H),8.25(bs,1H),7.99(d,J＝8.0 Hz,2H),7.69(dd,J＝7.4,1.4 Hz,1H),7.60(d,J＝8.4 Hz,2H),7.35(d,J＝8.4 Hz,1H),7.32(d,J＝2.5 Hz,1H),7.19(五重峰d,J＝7.2,2.3 Hz,2H),6.91(dd,J＝8.6,2.5 Hz,1H),4.79(s,2H),4.30(t,J＝5.3 Hz,2H),3.92(t,J＝5.5 Hz,3H),1.48(s,9H)。m/z：553.5,554.5(M＋ ,M＋1)。 N-(2-Amino-phenyl)-4-{[6-(2-chloro-ethoxy)-benzoquinazolyl-2-ylamino]-methyl}-benzamide (135) Step 1: [2-(4-([6-(2-Chloro-ethoxy)-benzothiazol-2-ylamino]-methyl)-benzylideneamino)-phenyl]-amine The title compound 130 (yield 43%) was obtained from the title compound (129) using the compound 129 as a starting material, according to the procedure of J. Med. Chem., 2002, 45 (6), 1300-1312. 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.59 (bs, 1H), 8.25 (bs, 1H), 7.99 (d, J = 8.0 Hz, 2H), 7.69 (dd, J = 7.4, 1.4 Hz, 1H), 7.60 (d, J = 8.4 Hz, 2H), 7.35 (d, J = 8.4 Hz, 1H), 7.32 (d, J = 2.5 Hz, 1H), 7.19 (five peaks d, J = 7.2, 2.3 Hz, 2H), 6.91 (dd, J = 8.6, 2.5 Hz, 1H), 4.79 (s, 2H), 4.30 (t, J = 5.3 Hz, 2H), 3.92 (t, J = 5.5 Hz, 3H), 1.48 (s, 9H). m/z: 553.5, 554.5 (M + , M + 1).
步驟2：N－(2－胺基－苯基)－4－{[6－(2－氯－乙氧基)－苯幷噻唑－2－基胺基]－甲基}－苯甲醯胺(135) 依照與流程28步驟5(實例68)對Boc***所述相同之程序自化合物130開始獲得標題化合物135(產率48%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.59(s,1H),8.39(t,J＝5.5 Hz,1H),7.92(d,J＝8.0 Hz,2H),7.46(d,J＝8.2 Hz,2H),7.35(d,J＝2.5 Hz,1H),7.27(d,J＝8.8 Hz,1H),7.13(d,J＝6.8 Hz,1H),6.94(td,J＝8.0,1.4 Hz,1H),6.83(dd,J＝8.8,6.1 Hz,1H),6.75(dd,J＝8.0,1.4 Hz,1H),6.57(t,J＝8.6 Hz,1H),4.88(s,2H),4.63(d,J＝6.1 Hz,2H),4.21(t,J＝5.1 Hz,2H),3.92(t,J＝5.3 Hz,2H)。mz：453.4,455.4(M＋ ,M＋1)。 Step 2: N-(2-Amino-phenyl)-4-{[6-(2-chloro-ethoxy)-benzothiazol-2-ylamino]-methyl}-benzamide (135) The title compound 135 (yield: 48%) was obtained from compound 130 starting from the same procedure as for the Boc cle. 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.59 (s, 1H), 8.39 (t, J = 5.5 Hz, 1H), 7.92 (d, J = 8.0 Hz, 2H), 7.46 (d, J = 8.2 Hz, 2H), 7.35 (d, J = 2.5 Hz, 1H), 7.27 (d, J = 8.8 Hz, 1H), 7.13 (d, J = 6.8 Hz, 1H), 6.94 (td, J = 8.0, 1.4 Hz, 1H), 6.83 (dd, J = 8.8, 6.1 Hz, 1H), 6.75 (dd, J = 8.0, 1.4 Hz, 1H), 6.57 (t, J = 8.6 Hz, 1H), 4.88 ( s, 2H), 4.63 (d, J = 6.1 Hz, 2H), 4.21 (t, J = 5.1 Hz, 2H), 3.92 (t, J = 5.3 Hz, 2H). Mz: 453.4, 455.4 (M + , M+1).
N－(2－胺基－苯基)－4－({6－[2－(4－甲基－六氫吡嗪－1－基)－乙氧基]－苯幷噻唑－2－基胺基}－甲基)－苯甲醯胺(136)步驟1：{2－[4－({6－[2－(4－甲基－六氫吡嗪－1－基)－乙氧基]－苯幷噻唑－2－基胺基}－甲基)－苯甲醯胺基}－苯基}－胺基甲酸第三丁酯(131)： 依照J.Med.Chem.,2002,45,(6),1300－1312所述之程序用化合物130作為原料獲得標題化合物131(產率91%)。1 H NMR：(丙酮－d6 )δ(ppm)：7.99(d,J＝8.2 Hz,2H),7.69(d,J＝7.2 Hz,1H),7.59(d,J＝8.6 Hz,2H),7.56(d,J＝2.0 Hz,1H),7.33(d,J＝8.8 Hz,1H),7.28(d,J＝2.5 Hz,1H),7.21(五重峰d,J＝7.2,1.2 Hz,2H),7.03－6.93(m,1H),6.87(dd,J＝8.8,2.7 Hz,1H),4.79(s,2H),4.11(t,J＝5.9 Hz,2H),2.75(t,J＝5.7 Hz,2H),2.67－2.51(m,4H),2.48－2.38(m,4H),2.21(s,3H),1.49(s,9H)。 N-(2-Amino-phenyl)-4-({6-[2-(4-methyl-hexahydropyrazin-1-yl)-ethoxy]-benzothiazol-2-ylamine Base}-methyl)-benzamide (136) Step 1: {2-[4-({6-[2-(4-Methyl-hexahydropyrazin-1-yl)-ethoxy] -Benzylthiazol-2-ylamino}-methyl)-benzylaminoamino}-phenyl}-aminocarboxylic acid tert-butyl ester (131): according to J. Med. Chem., 2002, 45, (6), the procedure described in 1300-1312, using Compound 130 as a starting material to give the title compound 131 (yield: 91%). 1 H NMR: (acetone-d 6 ) δ (ppm): 7.99 (d, J = 8.2 Hz, 2H), 7.69 (d, J = 7.2 Hz, 1H), 7.59 (d, J = 8.6 Hz, 2H) , 7.56 (d, J = 2.0 Hz, 1H), 7.33 (d, J = 8.8 Hz, 1H), 7.28 (d, J = 2.5 Hz, 1H), 7.21 (five peaks d, J = 7.2, 1.2 Hz) , 2H), 7.03-6.93 (m, 1H), 6.87 (dd, J = 8.8, 2.7 Hz, 1H), 4.79 (s, 2H), 4.11 (t, J = 5.9 Hz, 2H), 2.75 (t, J = 5.7 Hz, 2H), 2.67-2.51 (m, 4H), 2.48-2.38 (m, 4H), 2.21 (s, 3H), 1.49 (s, 9H).
步驟2：N－(2－胺基－苯基)－4－({6－[2－(4－甲基－六氫吡嗪－1－基)－乙氧基]－苯幷噻唑－2－基胺基}－甲基)－苯甲醯胺(136) 依照與流程28步驟5(實例68)對Boc***所述相同之程序自化合物131開始獲得標題化合物136(產率60%)。1 H NMR：(CDCl3 )δ(ppm)：7.97(d,J＝7.9 Hz,2H),7.58(d,J＝7.9 Hz,2H),7.51(d,J＝8.8 Hz,1H),7.42(d,J＝8.0 Hz,1H),7.22－7.17(m,2H),6.99－6.92(m,3H),4.78(s,2H),4.20－4.18(m,2H),2.97－2.87(m,8H),2.70－2.66(m,2H),2.61(s,3H)。m/z：517.5(MH＋ )。 Step 2: N-(2-Amino-phenyl)-4-({6-[2-(4-methyl-hexahydropyrazin-1-yl)-ethoxy]-benzothiazole-2 -Aminomethyl}-methyl)-benzimidamide (136) The title compound 136 (yield 60%) was obtained from compound 131 starting from the same procedure as described for the Boc ss. 1 H NMR: (CDCl 3 ) δ (ppm): 7.97 (d, J = 7.9 Hz, 2H), 7.58 (d, J = 7.9 Hz, 2H), 7.51 (d, J = 8.8 Hz, 1H), 7.42 (d, J=8.0 Hz, 1H), 7.22-7.17 (m, 2H), 6.99-6.92 (m, 3H), 4.78 (s, 2H), 4.20-4.18 (m, 2H), 2.97-2.87 (m , 8H), 2.70-2.66 (m, 2H), 2.61 (s, 3H). m/z: 517.5 (MH + ).
(2－嗎啉－4－基－乙基)－胺基甲酸2－[4－(2－胺基－苯胺甲醯基)－苄胺基]－苯幷噻唑－6－基酯(137)步驟1：[2－(4－{[6－(2－嗎啉－4－基－乙基胺甲醯氧基)－苯幷噻唑－2－基胺基]－甲基}－苯甲醯胺基)－苯基]－胺基甲酸第三丁酯(132)： N2 氣氛下向冷卻至－78℃的氯甲酸對硝基苯酯(171 mg，0.848 mmol)之THF(15 mL)溶液中加入Et3 N(236μL，1.70 mmol)。接著經套管添加中間體129(416 mg，0.848 mmol)之THF(4.2 mL)懸浮液。在－78℃下攪拌所得黃色混合物1小時且在0℃下攪拌其1.5小時，將其在40℃下加熱16小時且冷卻至室溫。接著添加純4－(2－胺乙基)嗎啉(119μL，0.848 mmol)且攪拌溶液4小時，藉由添加MeOH中止。將其攪拌30分鐘且加以濃縮。使用MeOH/DCM(3：97)由急驟層析法純化所得材料產生標題化合物132(165 mg，產率30%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.77(s,1H),8.64(bs,1H),8.56(t,J＝5.9 Hz,1H),7.90(d,J＝8.2 Hz,2H),7.60(t,J＝5.9 Hz,1H),7.50(d,J＝8.0 Hz,2H),7.50－7.48(m,2H),7.45(d,J＝2.3 Hz,1H),7.30(d,J＝8.8 Hz,1H),7.17(t,J＝7.6 Hz,1H),7.12(t,J＝7.8 Hz,1H),6.90(dd,J＝8.2,2.0 Hz,1H),4.67(d,J＝6.0 Hz,2H),3.56(t,J＝4.1 Hz,4H),3.31(t,J＝6.1 Hz,2H),3.16(q,J＝6.1 Hz,2H),2.39(t,J＝6.8 Hz,4H),1.42(s,9H)。m/z：647.7(MH＋ )。 (2-morpholin-4-yl-ethyl)-carbamic acid 2-[4-(2-amino-anilinyryl)-benzylamino]-benzothiazole-6-yl ester (137) Step 1: [2-(4-{[6-(2-Morpholin-4-yl-ethylaminemethyl methoxy)-benzothiazol-2-ylamino]-methyl}-benzamide Amino)-phenyl]-carbamic acid tert-butyl ester (132): p-nitrophenyl chloroformate (171 mg, 0.848 mmol) in THF (15 mL) cooled to -78 ° C under N 2 atmosphere. Et 3 N (236 μL, 1.70 mmol) was added to the solution. A suspension of Intermediate 129 (416 mg, 0.848 mmol) in THF (4.2 mL). The resulting yellow mixture was stirred at -78 °C for 1 hour and at 0 ° C for 1.5 hours, which was heated at 40 ° C for 16 hours and cooled to room temperature. Then pure 4-(2-aminoethyl)morpholine (119 μL, 0.848 mmol) was added and the solution was stirred for 4 h and quenched by MeOH. It was stirred for 30 minutes and concentrated. The material obtained was purified by flash chromatography using MeOH /EtOAc (EtOAc) 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.77 (s, 1H), 8.64 (bs, 1H), 8.56 (t, J = 5.9 Hz, 1H), 7.90 (d, J = 8.2 Hz, 2H), 7.60 (t, J = 5.9 Hz, 1H), 7.50 (d, J = 8.0 Hz, 2H), 7.50-7.48 (m, 2H), 7.45 (d, J = 2.3 Hz, 1H), 7.30 ( d, J = 8.8 Hz, 1H), 7.17 (t, J = 7.6 Hz, 1H), 7.12 (t, J = 7.8 Hz, 1H), 6.90 (dd, J = 8.2, 2.0 Hz, 1H), 4.67 ( d, J = 6.0 Hz, 2H), 3.56 (t, J = 4.1 Hz, 4H), 3.31 (t, J = 6.1 Hz, 2H), 3.16 (q, J = 6.1 Hz, 2H), 2.39 (t, J = 6.8 Hz, 4H), 1.42 (s, 9H). m/z: 647.7 (MH + ).
步驟2：(2－嗎啉－4－基－乙基)－胺基甲酸2－[4－(2－胺基－苯胺甲醯基)－苄胺基]－苯幷噻唑－6－基酯(137) 依照與流程28步驟5(實例68)對Boc***所述相同之程序自化合物132開始獲得標題化合物137(產率55%)。1 H NMR：(DMSO－d6 )：1 H NMR：(DMSO－d6 )：9.60(s,1H),8.56(t,J＝6.3 Hz,1H),7.93(d,J＝8.4 Hz,2H),7.61(t,J＝5.5 Hz,1H),7.47(d,J＝7.8 Hz,2H),7.46(d,J＝2.3 Hz,1H),7.31(d,J＝8.6 Hz,1H),7.14(d,J＝6.8 Hz,2H),6.95(t,J＝6.5 Hz,1H),6.92(dd,J＝8.8,2.5 Hz,1H),6.75(d,J＝7.8 Hz,1H),6.57(t,J＝7.4 Hz,1H),4.88(s,2H),4.66(d,J＝5.9 Hz,2H),3.57(t,J＝4.5 Hz,4H),3.33－3.31(m,2H),2.41－2.38(m,6H)。m/z：547.5(MH＋ )。 Step 2: (2-morpholin-4-yl-ethyl)-carbamic acid 2-[4-(2-amino-anilinomethyl)-benzylamino]-benzothiazole-6-yl ester (137) The title compound 137 (yield 55%) was obtained from compound 132 starting from the same procedure as described for the Boc cle. 1 H NMR: (DMSO-d 6 ): 1 H NMR: (DMSO-d 6 ): 9.60 (s, 1H), 8.56 (t, J = 6.3 Hz, 1H), 7.93 (d, J = 8.4 Hz, 2H), 7.61 (t, J = 5.5 Hz, 1H), 7.47 (d, J = 7.8 Hz, 2H), 7.46 (d, J = 2.3 Hz, 1H), 7.31 (d, J = 8.6 Hz, 1H) , 7.14 (d, J = 6.8 Hz, 2H), 6.95 (t, J = 6.5 Hz, 1H), 6.92 (dd, J = 8.8, 2.5 Hz, 1H), 6.75 (d, J = 7.8 Hz, 1H) , 6.57 (t, J = 7.4 Hz, 1H), 4.88 (s, 2H), 4.66 (d, J = 5.9 Hz, 2H), 3.57 (t, J = 4.5 Hz, 4H), 3.33 - 3.31 (m, 2H), 2.41-2.38 (m, 6H). m/z: 547.5 (MH + ).
N－(2－胺基－苯基)－4－{[6－(2－二甲胺基－乙氧基)－苯幷噻唑－2－基胺基]－甲基}－苯甲醯胺(138)步驟1：[2－(4－{[6－(2－二甲胺基－乙氧基)－苯幷噻唑－2－基胺基]－甲基}－苯甲醯胺基)－苯基]－胺基甲酸第三丁酯(133)： 在室溫下N2 氣氛下向化合物129(1.00 g，2.04 mmol)之THF(6.8 mL)懸浮液中相繼加入N,N－二甲基乙醇胺(225μL，2.24 mmol)及三苯基膦(696 mg，2.65 mmol)繼之以偶氮二羧酸二異丙酯(550μL，2.65 mmol)。放出熱量且混合物變成暗紅色。攪拌該混合物4小時，真空下移除THF且使暗色殘餘物在EtOAc與H2 O之間分溶。收集有機相且以HCl 1 N萃取之。分離且在劇烈攪拌下以飽和NaHCO3 中和酸性萃取物。形成白色沉澱，其經過濾收集產生標題化合物133(430 mg，產率37%)。1 H NMR：(丙酮－d6 )δ(ppm)：7.99(d,J＝8.4 Hz，2H),7.70(dd,J＝8.0,2.2 Hz,1H),7.59(d,J＝8.4 Hz,2H),7.56(d,J＝1.6 Hz,1H),7.33(d,J＝9.0 Hz,1H),7.27(d,J＝2.5 Hz,1H),7.19(五重峰d,J＝7.8,2.4 Hz,2H),6.87(dd,J＝8.8,2.5 Hz,1H),4.80(s,2H),4.08(t,J＝5.9 Hz,2H),2.67(t,J＝5.7 Hz,2H),2.27(s,6H),1.48(s,9H)。m/z：562.5(MH＋ )。 N-(2-Amino-phenyl)-4-{[6-(2-dimethylamino-ethoxy)-benzothiazol-2-ylamino]-methyl}-benzamide (138) Step 1: [2-(4-{[6-(2-Dimethylamino-ethoxy)-benzothiazol-2-ylamino]-methyl}-benzamide) -Phenyl]-carbamic acid tert-butyl ester (133): N,N-di was added successively to a suspension of compound 129 (1.00 g, 2.04 mmol) in THF (6.8 mL) at room temperature under N 2 atmosphere. Methylethanolamine (225 μL, 2.24 mmol) and triphenylphosphine (696 mg, 2.65 mmol) were followed by diisopropyl azodicarboxylate (550 μL, 2.65 mmol). The heat is released and the mixture turns dark red. The mixture was stirred for 4 hours, THF was removed under vacuum and the dark residue so partitioned between EtOAc and H 2 O. The organic phase was collected and extracted with HCl 1 N. Separated and under vigorous stirring with saturated NaHCO 3 and acidic extract. A white precipitate formed which was collected by filtration afforded the title compound 133 (430 mg, yield 37%). 1 H NMR: (acetone-d 6 ) δ (ppm): 7.99 (d, J = 8.4 Hz, 2H), 7.70 (dd, J = 8.0, 2.2 Hz, 1H), 7.59 (d, J = 8.4 Hz, 2H), 7.56 (d, J = 1.6 Hz, 1H), 7.33 (d, J = 9.0 Hz, 1H), 7.27 (d, J = 2.5 Hz, 1H), 7.19 (five peak d, J = 7.8, 2.4 Hz, 2H), 6.87 (dd, J = 8.8, 2.5 Hz, 1H), 4.80 (s, 2H), 4.08 (t, J = 5.9 Hz, 2H), 2.67 (t, J = 5.7 Hz, 2H) , 2.27 (s, 6H), 1.48 (s, 9H). m/z: 562.5 (MH + ).
步驟2：N－(2－胺基－苯基)－4－{[6－(2－二甲胺基－乙氧基)－苯幷噻唑－2－基胺基]－甲基}－苯甲醯胺(138) 依照與流程28步驟5(實例68)對Boc***所述相同之程序自化合物133開始獲得標題化合物138(產率82%)。1 H NMR：(CD3 OD)δ(ppm)：7.96(d,J＝8.2 Hz,2H),7.53(d,J＝8.2 Hz,2H),7.32(d,J＝8.8 Hz,1H),7.23(d,J＝2.5 Hz,1H),7.17(d,J＝9.0 Hz,1H),7.07(td,J＝9.0,1.6 Hz,1H),6.90(dd,J＝8.8,2.7 Hz,1H),6.89(dd,J＝6.5,1.6 Hz,1H),6.76(t,J＝6.5 Hz,1H),4.71(s,2H),4.10(t,J＝5.3 Hz,2H),2.79(t,J＝5.5 Hz,2H),2.36(s,6H)。m/z：462.5(MH＋ )。 Step 2: N-(2-Amino-phenyl)-4-{[6-(2-dimethylamino-ethoxy)-benzothiazol-2-ylamino]-methyl}-benzene The title compound 138 (yield 82%) was obtained from the title compound 138. 1 H NMR: (CD 3 OD) δ (ppm): 7.96 (d, J = 8.2 Hz, 2H), 7.53 (d, J = 8.2 Hz, 2H), 7.32 (d, J = 8.8 Hz, 1H), 7.23 (d, J = 2.5 Hz, 1H), 7.17 (d, J = 9.0 Hz, 1H), 7.07 (td, J = 9.0, 1.6 Hz, 1H), 6.90 (dd, J = 8.8, 2.7 Hz, 1H) ), 6.89 (dd, J = 6.5, 1.6 Hz, 1H), 6.76 (t, J = 6.5 Hz, 1H), 4.71 (s, 2H), 4.10 (t, J = 5.3 Hz, 2H), 2.79 (t , J = 5.5 Hz, 2H), 2.36 (s, 6H). m/z: 462.5 (MH + ).
N－(2－胺基－苯基)－4－{[6－(2－哌啶－1－基－乙氧基)－苯幷噻唑－2－基胺基]－甲基}－苯甲醯胺(139) 依照與實例76所述相同之程序(兩步反應順序)，但以1－哌啶乙醇代替N,N－二甲基乙醇胺獲得標題化合物139(經兩個步驟之產率52%)。1 H NMR：(CD3 OD)δ(ppm)：7.96(d,J＝8.4 Hz,2H),7.53(d,J＝8.2 Hz,2H),7.36(d,J＝8.8 Hz,1H),7.32(d,J＝2.5 Hz,1H),7.17(d,J＝8.0 Hz,1H),7.07(td,J＝6.1,1.2 Hz,1H),6.97(dd,J＝8.8,2.7 Hz,1H),6.90(d,J＝7.8 Hz,1H),6.77(t,J＝7.2 Hz,1H),4.71(s,2H),4.35(t,J＝4.9 Hz,2H),3.64－3.60(m,2H),3.56(t,J＝4.9 Hz,2H),3.10－3.01(m,2H),2.05－1.92(m,2H),1.90－1.81(m,4H)。m/z：502.5(MH＋ )。 N-(2-Amino-phenyl)-4-{[6-(2-piperidin-1-yl-ethoxy)-benzothiazol-2-ylamino]-methyl}-benzene The title compound 139 was obtained by the same procedure as described in Example 76 (two-step reaction sequence), but substituting 1-piperidineethanol for N,N-dimethylethanolamine (yield of two steps 52) %). 1 H NMR: (CD 3 OD) δ (ppm): 7.96 (d, J = 8.4 Hz, 2H), 7.53 (d, J = 8.2 Hz, 2H), 7.36 (d, J = 8.8 Hz, 1H), 7.32 (d, J = 2.5 Hz, 1H), 7.17 (d, J = 8.0 Hz, 1H), 7.07 (td, J = 6.1, 1.2 Hz, 1H), 6.97 (dd, J = 8.8, 2.7 Hz, 1H) ), 6.90 (d, J = 7.8 Hz, 1H), 6.77 (t, J = 7.2 Hz, 1H), 4.71 (s, 2H), 4.35 (t, J = 4.9 Hz, 2H), 3.64 - 3.60 (m) , 2H), 3.56 (t, J = 4.9 Hz, 2H), 3.10-3.01 (m, 2H), 2.05-1.92 (m, 2H), 1.90-1.81 (m, 4H). m/z: 502.5 (MH + ).
N－(2－胺基－苯基)－4－[(6－硝基－苯幷噻唑－2－基胺基)－甲基]－苯甲醯胺(142)步驟1：(2－{4－[(6－硝基－苯幷噻唑－2－基胺基)－甲基]－苯甲醯胺基}－苯基)－胺基甲酸第三丁酯(141)： 依照與流程3步驟2(實例12)對還原胺化反應所述相同之程序自化合物140與128(描述於專利申請案WO 03/024448中)開始獲得標題化合物141(產率66%)。1 H NMR：(CD3 OD)δ(ppm)：9.78(s,1H),9.28(bs,1H),8.71(d,J＝2.5 Hz,1H),8.64(bs,1H),8.09(dd,J＝9.0,2.5 Hz,1H),7.92(d,J＝8.2 Hz,2H),7.51(dd,J＝8.6,2.2 Hz,2H),7.46(d,J＝9.0,2H),7.17(td,J＝7.4,1.8 Hz,1H),7.12(td,J＝7.1,1.8 Hz,1H),4.75(bs,2H),1.42(s,9H)。m/z：542.2(M＋Na)。 N-(2-Amino-phenyl)-4-[(6-nitro-benzothiazol-2-ylamino)-methyl]-benzamide (142) Step 1: (2-{ 4-[(6-Nitro-benzoquinazolyl-2-ylamino)-methyl]-benzylaminoamido}-phenyl)-aminocarboxylic acid tert-butyl ester (141): according to Scheme 3 Step 2 (Example 12) The same procedure as described for the reductive amination reaction from compound 140 and 128 (described in patent application WO 03/024448) afforded the title compound 141 (yield 66%). 1 H NMR: (CD 3 OD) δ (ppm): 9.78 (s, 1H), 9.28 (bs, 1H), 8.71 (d, J = 2.5 Hz, 1H), 8.64 (bs, 1H), 8.09 (dd) , J=9.0, 2.5 Hz, 1H), 7.92 (d, J=8.2 Hz, 2H), 7.51 (dd, J=8.6, 2.2 Hz, 2H), 7.46 (d, J=9.0, 2H), 7.17 ( Td, J = 7.4, 1.8 Hz, 1H), 7.12 (td, J = 7.1, 1.8 Hz, 1H), 4.75 (bs, 2H), 1.42 (s, 9H). m/z: 542.2 (M+Na).
步驟2：N－(2－胺基－苯基)－4－[(6－硝基－苯幷噻唑－2－基胺基)－甲基]－苯甲醯胺(142) 依照與流程28步驟5(實例68)對Boc***所述相同之程序以化合物141作為原料獲得標題化合物142(產率98%)。1 H NMR：(DMSO－d6 )：10.06(s,1H),9.30(bs,1H),8.71(d,J＝2.3 Hz,1H),8.09(dd,J＝9.0,2.3 Hz,1H),9.97(d,J＝8.2 Hz,2H),7.51(d,J＝8.2 Hz,2H),7.46(d,J＝8.8 Hz,1H),7.30(d,J＝7.0 Hz,1H),7.17(t,J＝7.8 Hz,1H),7.10(d,J＝7.8 Hz,1H),7.03(t,J＝7.2 Hz,1H),4.75(d,J＝5.5 Hz,2H)。m/z：420.5(MH＋ )。 Step 2: N-(2-Amino-phenyl)-4-[(6-nitro-benzothiazol-2-ylamino)-methyl]-benzamide (142) according to Scheme 28 Step 5 (Example 68) The title compound 142 (yield: 98%) was obtained from Compound 141. 1 H NMR: (DMSO-d 6 ): 10.06 (s, 1H), 9.30 (bs, 1H), 8.71 (d, J = 2.3 Hz, 1H), 8.09 (dd, J = 9.0, 2.3 Hz, 1H) , 9.97 (d, J = 8.2 Hz, 2H), 7.51 (d, J = 8.2 Hz, 2H), 7.46 (d, J = 8.8 Hz, 1H), 7.30 (d, J = 7.0 Hz, 1H), 7.17 (t, J = 7.8 Hz, 1H), 7.10 (d, J = 7.8 Hz, 1H), 7.03 (t, J = 7.2 Hz, 1H), 4.75 (d, J = 5.5 Hz, 2H). m/z: 420.5 (MH + ).
4－[(6－胺基－苯幷噻唑－2－基胺基)－甲基]－N－(2－胺基－苯基)－苯甲醯胺(144)步驟1：(2－{4－[(6－胺基－苯幷噻唑－2－基胺基)－甲基]－苯甲醯胺基}－苯基)－胺基甲酸第三丁酯(143)： 向溶於THF/MeOH/H2 O(10 mL/10 mL/10 mL)混合物中之化合物141(200 mg，0.385 mmol)懸浮液中相繼加入二水合氯化錫(II)(1.35 g，8.46 mmol)及乙酸銨(1.09 g，14.12 mmol)。將混合物回流2天，濾除錫鹽且在真空下濃縮濾液。使殘餘物在EtOAc與H2 O之間分溶(加入鹽水破乳)。相繼以NaHCO3 飽和水溶液及鹽水洗滌該有機相，使其經MgSO4 乾燥，且在真空下濃縮產生標題化合物143(145 mg，產率77%)。1 H NMR：(DMSO－d6 )δ(ppm)：12.89(bs,1H),10.79(s,1H),8.12(d,J＝2.0 Hz,1H),8.05(d,J＝8.8 Hz,2H),7.90－7.68(m,3H),7.62(d,J＝8.4 Hz,2H),7.48(bs,1H),7.21(dd,J＝4.9,3.7 Hz,1H),4.65(s,2H)。m/z：490.5(MH＋ )。 4-[(6-Amino-benzoquinazolyl-2-ylamino)-methyl]-N-(2-amino-phenyl)-benzamide (144) Step 1: (2-{ 4-[(6-Amino-benzoquinazolyl-2-ylamino)-methyl]-benzylaminoamido}-phenyl)-aminocarboxylic acid tert-butyl ester (143): soluble in THF Addition of a solution of compound 141 (200 mg, 0.385 mmol) in a mixture of /MeOH/H 2 O (10 mL / 10 mL / 10 mL) in a mixture of tin(II) chloride (1.35 g, 8.46 mmol) and acetic acid Ammonium (1.09 g, 14.12 mmol). The mixture was refluxed for 2 days, the tin salt was filtered off and the filtrate was concentrated in vacuo. The residue was partitioned between EtOAc and H 2 O partitioned (brine was added demulsification). Washed successively with saturated aqueous NaHCO 3 and brine and the organic phase was dried over it MgSO 4, and concentrated in vacuo to produce the title compound 143 (145 mg, 77% yield). 1 H NMR: (DMSO-d 6 ) δ (ppm): 12.89 (bs, 1H), 10.79 (s, 1H), 8.12 (d, J = 2.0 Hz, 1H), 8.05 (d, J = 8.8 Hz, 2H), 7.90-7.68 (m, 3H), 7.62 (d, J = 8.4 Hz, 2H), 7.48 (bs, 1H), 7.21 (dd, J = 4.9, 3.7 Hz, 1H), 4.65 (s, 2H) ). m/z: 490.5 (MH + ).
步驟2：4－[(6－胺基－苯幷噻唑－2－基胺基)－甲基]－N－(2－胺基－苯基)－苯甲醯胺(144) 依照與流程28步驟5(實例68)對Boc***所述相同之程序用化合物143作為原料獲得標題化合物144(產率58%)。1 H NMR：(DMSO－d6 )：9.58(s,1H),8.09(t,J＝5.9 Hz,1H),7.91(d,J＝8.0 Hz,2H),7.45(d,J＝8.4 Hz,2H),7.13(d,J＝7.0 Hz,1H),7.05(d,J＝8.6 Hz,1H),6.94(t,J＝6.8 Hz,1H),6.81(d,J＝2.2 Hz,1H),6.75(dd,J＝6.7,1.2 Hz,1H),6.57(t,J＝6.5 Hz,1H),6.48(dd,J＝8.4,2.2 Hz,1H),5.19(s,2H),4.81(s,2H),4.58(d,J＝5.9 Hz,2H)。m/z：390.5(MH＋ )。 Step 2: 4-[(6-Amino-benzoquinazolyl-2-ylamino)-methyl]-N-(2-amino-phenyl)-benzamide (144) according to Scheme 28 Step 5 (Example 68) The title compound 144 (yield: 58%). 1 H NMR: (DMSO-d 6 ): 9.58 (s, 1H), 8.09 (t, J = 5.9 Hz, 1H), 7.91 (d, J = 8.0 Hz, 2H), 7.45 (d, J = 8.4 Hz) , 2H), 7.13 (d, J = 7.0 Hz, 1H), 7.05 (d, J = 8.6 Hz, 1H), 6.94 (t, J = 6.8 Hz, 1H), 6.81 (d, J = 2.2 Hz, 1H) ), 6.75 (dd, J = 6.7, 1.2 Hz, 1H), 6.57 (t, J = 6.5 Hz, 1H), 6.48 (dd, J = 8.4, 2.2 Hz, 1H), 5.19 (s, 2H), 4.81 (s, 2H), 4.58 (d, J = 5.9 Hz, 2H). m / z: 390.5 (MH + ).
N－(2－胺基－苯基)－4－[(5,6－二氟－苯幷噻唑－2－基胺基)－甲基]－苯甲醯胺(147)步驟1：5,6－二氟－苯幷噻唑－2－基胺(145)： 依照J.Het.Chem,1971,8(309－310)所述之程序自4,5－二氟苯胺開始獲得標題化合物145(產率95%)。1 H NMR：(DMSO－d6 )δ(ppm)：7.78(dd,J＝10.6,8.6 Hz,1H),7.61(s,2H),7.32(dd,J＝11.9,7.2 Hz,1H)。m/z：337.5(M＋Na＋ )。 N-(2-Amino-phenyl)-4-[(5,6-difluoro-benzothiazol-2-ylamino)-methyl]-benzamide (147) Step 1: 5, 6-Difluoro-benzothiazol-2-ylamine (145): The title compound 145 was obtained from 4,5-difluoroaniline according to the procedure of J. Het. Chem., 1971, 8 (309-310). Yield 95%). 1 H NMR: (DMSO-d 6) δ (ppm): 7.78 (dd, J = 10.6,8.6 Hz, 1H), 7.61 (s, 2H), 7.32 (dd, J = 11.9,7.2 Hz, 1H). m/z: 337.5 (M+Na + ).
步驟2：4－[(5,6－二氟－苯幷噻唑－2－基胺基)－甲基]－苯甲酸(146)： 依照與流程3步驟2(實例12)對還原胺化反應所述相同之程序自化合物145開始獲得標題化合物146(產率63%)。1 H NMR：(DMSO－d6 )δ(ppm)：8.72(t,J＝5.9 Hz,1H),7.89(d,J＝8.4 Hz,2H),7.82(dd,J＝10.4,8.0 Hz,1H),7.44(d,J＝8.4 Hz,2H),7.40(dd,J＝11.9,7.4 Hz,1H),4.65(d,J＝5.7 Hz,2H)。m/z：315.2(MH＋ )。 Step 2: 4-[(5,6-Difluoro-benzothiazol-2-ylamino)-methyl]-benzoic acid (146): Reductive amination according to Step 2 (Example 12) of Scheme 3 The title compound 146 (yield 63%) was obtained from compound 145. 1 H NMR: (DMSO-d 6 ) δ (ppm): 8.72 (t, J = 5.9 Hz, 1H), 7.89 (d, J = 8.4 Hz, 2H), 7.82 (dd, J = 10.4, 8.0 Hz, 1H), 7.44 (d, J = 8.4 Hz, 2H), 7.40 (dd, J = 11.9, 7.4 Hz, 1H), 4.65 (d, J = 5.7 Hz, 2H). m/z: 315.2 (MH + ).
步驟3：N－(2－胺基－苯基)－4－[(5,6－二氟－苯幷噻唑－2－基胺基)－甲基]－苯甲醯胺(147)： 依照與流程1步驟5(實例1)對BOP偶合反應所述相同之程序自化合物146開始獲得標題化合物147(產率32%)。1 H NMR：(DMSO－d6 )：9.59(s,1H),8.73(t,J＝5.9 Hz,1H),7.93(d,J＝8.2 Hz,2H),7.83(dd,J＝10.4,8.0 Hz,1H),7.45(d,J＝8.2 Hz,2H),7.40(dd,J＝11.9,7.2 Hz,1H),7.13(d,J＝7.8 Hz,1H),6.94(td,J＝7.8,1.4 Hz,1H),6.75(dd,J＝7.8,1.4 Hz,1H),6.57(td,J＝7.6,1.2 Hz,1H),4.87(s,2H),4.65(d,J＝5.9 Hz,2H)。m/z：411.4(MH＋ )。 Step 3: N-(2-Amino-phenyl)-4-[(5,6-difluoro-benzothiazol-2-ylamino)-methyl]-benzamide (147): The title compound 147 (yield 32%) was obtained from compound 146 starting from the same procedure as for the </ br> 1 H NMR: (DMSO-d 6 ): 9.59 (s, 1H), 8.73 (t, J = 5.9 Hz, 1H), 7.93 (d, J = 8.2 Hz, 2H), 7.83 (dd, J = 10.4, 8.0 Hz, 1H), 7.45 (d, J = 8.2 Hz, 2H), 7.40 (dd, J = 11.9, 7.2 Hz, 1H), 7.13 (d, J = 7.8 Hz, 1H), 6.94 (td, J = 7.8, 1.4 Hz, 1H), 6.75 (dd, J = 7.8, 1.4 Hz, 1H), 6.57 (td, J = 7.6, 1.2 Hz, 1H), 4.87 (s, 2H), 4.65 (d, J = 5.9 Hz, 2H). m/z: 411.4 (MH + ).
N－(2－胺基－苯基)－4－{6－[3－(2－嗎啉－4－基－乙基)－脲基]－苯幷噻唑－2－基硫基甲基}－苯甲醯胺(157)步驟1：4－{6－[3－(2－嗎啉－4－基－乙基)－脲基]－苯幷噻唑－2－基硫基甲基}－苯甲酸甲酯(149)： 依照與流程32步驟1(實例75)對形成胺基甲酸酯所述相同之程序，但以化合物148(描述於專利申請案WO 03/024448中)代替化合物129獲得標題化合物149(產率70%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.28(bs,1H),8.18(d,J＝2.3 Hz,1H),7.90(d,J＝8.2 Hz,2H),7.77(d,J＝8.6 Hz,1H),7.61(d,J＝8.2 Hz,2H),7.42(dd,J＝8.8,2.2 Hz,1H),4.68(s,2H),3.82(s,3H),3.59－3.58(m,4H),3.33－3.32(m,2H),3.21(q,J＝6.1 Hz,2H),2.38－2.37(m,4H)。m/z：487.4(MH＋ )。 N-(2-Amino-phenyl)-4-{6-[3-(2-morpholin-4-yl-ethyl)-ureido]-benzoquinone-2-ylthiomethyl} -benzimidamide (157) Step 1: 4-{6-[3-(2-morpholin-4-yl-ethyl)-ureido]-benzothiazol-2-ylthiomethyl}- benzoate (149): Following the same procedures of the urethane and processes 32 step 1 (example 75) formed, but in place of compound 148 compound 129 (described in the Patent application WO 03/024448) The title compound 149 (yield 70%) was obtained. 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.28 (bs, 1H), 8.18 (d, J = 2.3 Hz, 1H), 7.90 (d, J = 8.2 Hz, 2H), 7.77 (d, J = 8.6 Hz, 1H), 7.61 (d, J = 8.2 Hz, 2H), 7.42 (dd, J = 8.8, 2.2 Hz, 1H), 4.68 (s, 2H), 3.82 (s, 3H), 3.59- 3.58 (m, 4H), 3.33 - 3.32 (m, 2H), 3.21 (q, J = 6.1 Hz, 2H), 2.38-2.37 (m, 4H). m/z: 487.4 (MH + ).
步驟2：4－{6－[3－(2－嗎啉－4－基－乙基)－脲基]－苯幷噻唑－2－基硫基甲基}－苯甲酸甲酯(153)： 依照與流程1步驟4(實例1)對水解所述相同之程序，用化合物149作為原料獲得標題化合物153(產率50%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.75(bs,1H),8.22(d,J＝2.2 Hz,1H),7.90(d,J＝8.4 Hz,2H),7.76(d,J＝8.8 Hz,1H),7.61(d,J＝8.4 Hz,2H),7.55(d,J＝6.3 Hz,1H),7.49(dd,J＝8.8,2.2 Hz,1H),4.68(s,2H),3.58(t,J＝4.3 Hz,4H),3.34－3.32(m,2H),3.21(q,J＝5.9 Hz,2H),2.38(t,J＝6.3 Hz,4H)。m/z：473.4(MH＋ )。 Step 2: 4-{6-[3-(2-Morpholin-4-yl-ethyl)-ureido]-benzoquinone-2-ylthiomethyl}-benzoic acid methyl ester (153): The title compound 153 (yield 50%) was obtained from Compound 149 (m.). 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.75 (bs, 1H), 8.22 (d, J = 2.2 Hz, 1H), 7.90 (d, J = 8.4 Hz, 2H), 7.76 (d, J = 8.8 Hz, 1H), 7.61 (d, J = 8.4 Hz, 2H), 7.55 (d, J = 6.3 Hz, 1H), 7.49 (dd, J = 8.8, 2.2 Hz, 1H), 4.68 (s, 2H), 3.58 (t, J = 4.3 Hz, 4H), 3.34 - 3.32 (m, 2H), 3.21 (q, J = 5.9 Hz, 2H), 2.38 (t, J = 6.3 Hz, 4H). m/z: 473.4 (MH + ).
步驟3：N－(2－胺基－苯基)－4－{6－[3－(2－嗎啉－4－基－乙基)－胺基]－苯幷噻唑－2－基硫基甲基}－苯甲醯胺(157) 依照與流程1步驟5(實例1)對BOP偶合所述相同之程序以化合物153作為原料獲得標題化合物157(產率26%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.59(s,1H),8.84(s,1H),8.13(d,J＝2.2 Hz,1H),7.90(d,J＝8.2 Hz,2H),7.71(d,J＝8.6 Hz,1H),7.58(d,J＝8.2 Hz,2H),7.30(dd,J＝8.8,2.2 Hz 1H),7.12(d,J＝7.0 Hz,1H),6.94(t,J＝7.0 Hz,1H),6.74(dd,J＝8.1,1.5 Hz,1H),6.56(t,J＝7.4 Hz,1H),6.14(t,J＝4.9 Hz,1H),4.88(bs,2H),4.66(s,2H)3.58(t,J＝4.5 Hz,4H),3.31－3.30(m,2H),3.21(q,J＝5.7 Hz,2H),2.38(t,J＝6.3 Hz,4H)。m/z：563.5(MH＋ )。 Step 3: N-(2-Amino-phenyl)-4-{6-[3-(2-morpholin-4-yl-ethyl)-amino]-benzoquinone-2-ylthio Methyl}-benzamide (157) The title compound 157 (yield: 26%) was obtained from the compound 153 as the starting material in the same procedure as in the procedure of the procedure of the procedure. 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.59 (s, 1H), 8.84 (s, 1H), 8.13 (d, J = 2.2 Hz, 1H), 7.90 (d, J = 8.2 Hz, 2H), 7.71 (d, J = 8.6 Hz, 1H), 7.58 (d, J = 8.2 Hz, 2H), 7.30 (dd, J = 8.8, 2.2 Hz 1H), 7.12 (d, J = 7.0 Hz, 1H) ), 6.94 (t, J = 7.0 Hz, 1H), 6.74 (dd, J = 8.1, 1.5 Hz, 1H), 6.56 (t, J = 7.4 Hz, 1H), 6.14 (t, J = 4.9 Hz, 1H) ), 4.88 (bs, 2H), 4.66 (s, 2H) 3.58 (t, J = 4.5 Hz, 4H), 3.31-3.30 (m, 2H), 3.21 (q, J = 5.7 Hz, 2H), 2.38 ( t, J = 6.3 Hz, 4H). m/z: 563.5 (MH + ).
N－(2－胺基－苯基)－4－[6－(2－二甲胺基－乙醯胺基)－苯幷噻唑－2－基硫基甲基]－苯甲醯胺(158)步驟1：4－[6－(2－二甲胺基－乙醯胺基)－苯幷噻唑－2－基硫基甲基]－苯甲酸甲酯(150)： 向化合物148(描述於專利申請案WO 03/024448中)(701 mg，2.12 mmol)及Me2 NCH2 COCl．HCl(670 mg，4.24 mmol)之CH3 CN之懸浮液加入NaHCO3 (356 mg，4.24 mmol)繼之以加入Et3 N(295μl，2.12 mmol)。在室溫下攪拌該混合物24小時，將其真空下濃縮且使殘餘物在DCM與H2 O之間分溶。收集含水層，將其以NaHCO3 中和且以新鮮DCM萃取，經Na2 SO4 乾燥且真空下濃縮之。由急驟層析法經矽膠純化殘餘物產生標題化合物150(485 mg，產率55%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.95(s,1H),8.41(d,J＝2.0 Hz,1H),7.91(d,J＝8.2 Hz,2H),7.79(d,J＝8.8 Hz,1H),7.63(d,J＝8.2 Hz,2H),7.63(dd,J＝8.8,2.1 Hz,1H),4.71(s,2H),3.84(s,3H),3.11(s,2H),2.30(s,6H)。m/z：416.4(MH＋ )。 N-(2-Amino-phenyl)-4-[6-(2-dimethylamino-acetamido)-benzoquinone-2-ylthiomethyl]-benzamide (158 Step 1: 4-[6-(2-Dimethylamino-acetamido)-benzoquinone-2-ylthiomethyl]-benzoic acid methyl ester (150): to compound 148 (described in Patent application WO 03/024448) (701 mg, 2.12 mmol) and Me 2 NCH 2 COCl. HCl (670 mg, 4.24 mmol) of CH 3 CN was added a suspension of NaHCO 3 (356 mg, 4.24 mmol ) was added followed by Et 3 N (295μl, 2.12 mmol ). The mixture was stirred at room temperature for 24 hours, it was concentrated under vacuo and the residue was partitioned between DCM and H 2 O. The aqueous layer was collected, to which NaHCO 3 and extracted with fresh DCM and, over the next 2 SO 4 dried, and concentrated in vacuo of Na. The residue was purified by flash chromatography eluting elut elut elut elut 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.95 (s, 1H), 8.41 (d, J = 2.0 Hz, 1H), 7.91 (d, J = 8.2 Hz, 2H), 7.79 (d, J = 8.8 Hz, 1H), 7.63 (d, J = 8.2 Hz, 2H), 7.63 (dd, J = 8.8, 2.1 Hz, 1H), 4.71 (s, 2H), 3.84 (s, 3H), 3.11 ( s, 2H), 2.30 (s, 6H). m/z: 416.4 (MH + ).
步驟2：4－[6－(2－二甲胺基－乙醯胺基)－苯幷噻唑－2－基硫基甲基]－苯甲酸(154)： 依照與流程1步驟4(實例1)對水解所述相同之程序用化合物150作為原料獲得標題化合物154(產率78%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.95(s,1H),8.41(d,J＝2.0 Hz,1H),7.86(d,J＝8.2 Hz,2H),7.79(d,J＝8.8 Hz,1H),7.63(dd,J＝9.0,2.0 Hz,1H),7.55(d,J＝8.2 Hz,2H),4.68(s,2H),3.11(s,2H),2.30(s,6H)。m/z：402.4(MH＋ )。 Step 2: 4-[6-(2-Dimethylamino-acetamido)-benzoquinone-2-ylthiomethyl]-benzoic acid (154): according to step 4 of Scheme 1 (Example 1 The title compound 154 (yield: 78%) was obtained from the title compound 152. 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.95 (s, 1H), 8.41 (d, J = 2.0 Hz, 1H), 7.86 (d, J = 8.2 Hz, 2H), 7.79 (d, J = 8.8 Hz, 1H), 7.63 (dd, J = 9.0, 2.0 Hz, 1H), 7.55 (d, J = 8.2 Hz, 2H), 4.68 (s, 2H), 3.11 (s, 2H), 2.30 ( s, 6H). m/z: 402.4 (MH + ).
步驟3：N－(2－胺基－苯基)－4－[6－(2－二甲胺基－乙醯胺基)－苯幷噻唑－2－基硫基甲基]－苯甲醯胺(158) 依據與流程1步驟5(實例1)對BOP偶合所述相同之程序用化合物154作為原料獲得標題化合物158(產率28%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.93(s,1H),9.59(s,1H),8.39(d,J＝2.0 Hz,1H),7.90(d,J＝8.0 Hz,2H),7.79(d,J＝9.0 Hz,1H),7.62(dd,J＝8.8,2.2 Hz,1H),7.60(d,J＝8.2 Hz,2H),7.12(d,J＝7.6 Hz,1H),6.94(t,J＝8.0 Hz,1H),6.74(dd,J＝8.0,1.6 Hz,1H),6.56(t,J＝7.5 Hz,1H),4.88(s,2H),4.69(s,2H),3.09(s,2H),2.28(s,6H)。HRMS：m/z：491.1455±0.0014(M＋)。 Step 3: N-(2-Amino-phenyl)-4-[6-(2-dimethylamino-acetamido)-benzoquinone-2-ylthiomethyl]-benzamide The title compound 158 (yield 28%) was obtained from the title compound 158 (yield 28%). 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.93 (s, 1H), 9.59 (s, 1H), 8.39 (d, J = 2.0 Hz, 1H), 7.90 (d, J = 8.0 Hz, 2H), 7.79 (d, J = 9.0 Hz, 1H), 7.62 (dd, J = 8.8, 2.2 Hz, 1H), 7.60 (d, J = 8.2 Hz, 2H), 7.12 (d, J = 7.6 Hz, 1H), 6.94 (t, J = 8.0 Hz, 1H), 6.74 (dd, J = 8.0, 1.6 Hz, 1H), 6.56 (t, J = 7.5 Hz, 1H), 4.88 (s, 2H), 4.69 ( s, 2H), 3.09 (s, 2H), 2.28 (s, 6H). HRMS: m/z: 491.1455 ± 0.0014 (M+).
N－(2－胺基－苯基)－4－[6－(二甲胺基－亞甲胺基)－苯幷噻唑－2－基硫基甲基]－苯甲醯胺(159)步驟1：4－[6－(二甲胺基－亞甲胺基)－苯幷噻唑－2－基硫基甲基]－苯甲酸甲酯(151)： 在N2 氣氛下向氯甲酸三氯甲酯(74μL，608 mmol)之THF(2 mL)預冷(－78℃)溶液中經由套管加入溶於THF及DMF(分別為3.5 mL，0.5 mL)之混合物中的化合物148(描述於專利申請案WO 03/024448中)(201 mg，608 mmol)溶液，隨後添加Et3 N(169μL，1.22 mmol)。將該溶液在－78℃下攪拌1小時且在0℃下攪拌2小時並溫至室溫隔夜。真空下移除溶劑，使殘餘物在H2 O與DCM/MeOH(9：1)之混合物間分溶，經MgSO4 乾燥且在真空下濃縮產生標題化合物151(136 mg，產率58%)。1 H NMR：(DMSO－d6 )δ(ppm)：7.92(d,J＝8.4 Hz,2H),7.81(s,1H),7.69(d,J＝8.6 Hz,1H),7.62(d,J＝8.2 Hz,2H),7.46(d,J＝2.2 Hz,1H),7.04(dd,J＝8.6,2.2 Hz,1H),4.68(s,2H),3.84(s,3H),3.04(bs,3H),2.95(bs,3H)。m/z：386.4(MH＋ )。 N-(2-Amino-phenyl)-4-[6-(dimethylamino-methyleneamino)-benzoquinone-2-ylthiomethyl]-benzamide (159) step 1:4-[6-(Dimethylamino-methyleneamino)-benzothiazol-2-ylthiomethyl]-benzoic acid methyl ester (151): trichlorochloroformic acid under N 2 atmosphere Methyl ester (74 μL, 608 mmol) in THF (2 mL) in EtOAc (2 mL) EtOAc (EtOAc, EtOAc) A solution (201 mg, 608 mmol) of the patent application WO 03/024448 followed by the addition of Et 3 N (169 μL, 1.22 mmol). The solution was stirred at -78 °C for 1 hour and at 0 °C for 2 hours and warmed to room temperature overnight. The solvent is removed in vacuo, the residue was H 2 O and DCM / MeOH: partitioned between a mixture of (91), the dried over MgSO 4 and concentrated under vacuo to give the title compound 151 (136 mg, 58% yield) . 1 H NMR: (DMSO-d 6 ) δ (ppm): 7.92 (d, J = 8.4 Hz, 2H), 7.81 (s, 1H), 7.69 (d, J = 8.6 Hz, 1H), 7.62 (d, J = 8.2 Hz, 2H), 7.46 (d, J = 2.2 Hz, 1H), 7.04 (dd, J = 8.6, 2.2 Hz, 1H), 4.68 (s, 2H), 3.84 (s, 3H), 3.04 ( Bs, 3H), 2.95 (bs, 3H). m/z: 386.4 (MH + ).
步驟2：4－[6－(二甲胺基－亞甲胺基)－苯幷噻唑－2－基硫基甲基]－苯甲酸(155)： 依照與流程1步驟4(實例1)對水解所述相同之程序以化合物151作為原料獲得標題化合物155(產率45%)。1 H NMR：(DMSO－d6 )δ(ppm)：7.89(d,J＝8.2 Hz,2H),7.80(s,1H),7.69(d,J＝8.6 Hz,1H),7.59(d,J＝8.2 Hz,2H),7.45(d,J＝2.2 Hz,1H),7.04(dd,J＝8.6,2.2 Hz,1H),4.67(s,2H),3.03(bs,3H),2.94(bs,3H)。m/z：372.3(MH＋ )。 Step 2: 4-[6-(Dimethylamino-methyleneamino)-benzothiazol-2-ylthiomethyl]-benzoic acid (155): according to step 4 of Example 1 (Example 1) Hydrolysis of the same procedure The title compound 155 (yield 45%) was obtained from compound 151. 1 H NMR: (DMSO-d 6 ) δ (ppm): 7.89 (d, J = 8.2 Hz, 2H), 7.80 (s, 1H), 7.69 (d, J = 8.6 Hz, 1H), 7.59 (d, J = 8.2 Hz, 2H), 7.45 (d, J = 2.2 Hz, 1H), 7.04 (dd, J = 8.6, 2.2 Hz, 1H), 4.67 (s, 2H), 3.03 (bs, 3H), 2.94 ( Bs, 3H). m/z: 372.3 (MH + ).
步驟3：N－(2－胺基－苯基)－4－[6－(二甲胺基－亞甲胺基)－苯幷噻唑－2－基硫基甲基]－苯甲醯胺(159) 依照與流程1步驟5(實例1)對BOP偶合所述相同之程序用化合物155作為原料獲得標題化合物159(產率25%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.60(s,1H),7.91(d,J＝8.2 Hz,2H),7.79(s,1H),7.69(d,J＝8.6 Hz,1H),7.60(d,J＝8.2 Hz,2H),7.44(d,J＝2.2 Hz,1H),7.13(d,J＝8.0 Hz,1H),7.03(dd,J＝8.6,2.3 Hz,1H),6.95(t,J＝7.0 Hz,1H),6.75(d,J＝9.2 Hz,1H),6.57(t,J＝7.4 Hz,1H),4.89(s,2H),4.67(s,2H),3.02(s,3H),2.93(s,3H)。m/z：462.5(MH＋ )。 Step 3: N-(2-Amino-phenyl)-4-[6-(dimethylamino-methyleneamino)-benzoquinone-2-ylthiomethyl]-benzamide ( 159) to obtain the title compound 159 (yield 25%) in accordance with Scheme 1 step 5 (example 1) the same procedure BOP coupling of the compound 155 as starting material. 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.60 (s, 1H), 7.91 (d, J = 8.2 Hz, 2H), 7.79 (s, 1H), 7.69 (d, J = 8.6 Hz, 1H), 7.60 (d, J = 8.2 Hz, 2H), 7.44 (d, J = 2.2 Hz, 1H), 7.13 (d, J = 8.0 Hz, 1H), 7.03 (dd, J = 8.6, 2.3 Hz, 1H), 6.95 (t, J = 7.0 Hz, 1H), 6.75 (d, J = 9.2 Hz, 1H), 6.57 (t, J = 7.4 Hz, 1H), 4.89 (s, 2H), 4.67 (s, 2H), 3.02 (s, 3H), 2.93 (s, 3H). m/z: 462.5 (MH + ).
N－(2－胺基－苯基)－4－{6－1N－(2－胺基－苯基)－4－苯甲醯胺]－苯幷噻唑－2－基硫基甲基}－苯甲醯胺(160)步驟1：N－(4－甲基苯甲酸甲酯)－苯幷噻唑－2－基硫基甲基}－苯甲酸甲酯(152)： 向化合物148(9.52 g，28.8 mmol)之DMF(30 mL)溶液中加入DCM(130 mL)及(4－溴甲基)苯甲酸甲酯(6.60 g，28.8 mmol)且在室溫下攪拌該混合物16小時。真空下濃縮溶劑且使所得固體在EtOAc與H2 O之間分溶。以HCl 1 N及鹽水洗滌有機層，使其經MgSO4 乾燥且在真空下濃縮之。使用EtOAc/Hex(45：55)由急驟層析法純化粗材料隨後使用MeOH/DCM(2：98)經Biotage預裝填矽膠柱進行純化，且在CHCl3 與Et2 O之混合物中結晶產生標題化合物152(2.66 g，產率19%)。1 H NMR：(DMsO－d6 )δ(ppm)：7.89(d,J＝8.0 Hz,2H),7.87(d,J＝7.8 Hz,2H),7.55(d,J＝8.4 Hz,2H),7.54(d,J＝8.8 Hz,1H),7.47(d,J＝8.4 Hz,2H),6.93(d,J＝2.3 Hz,1H),6.77(dd,J＝8.8,2.5 Hz,1H),6.70(t,J＝6.1 Hz,1H),4.58(s,2H),4.38(d,J＝6.3 Hz,2H),3.81(s,3H),3.81(s,3H)。m/z：479.4(MH＋ )。 N-(2-Amino-phenyl)-4-{6-1N-(2-amino-phenyl)-4-benzylideneamine]-benzoquinone-2-ylthiomethyl}- Benzamide (160) Step 1: N-(4-methylbenzoic acid methyl ester)-benzoquinone-2-ylthiomethyl}-benzoic acid methyl ester (152): to compound 148 (9.52 g DCM (130 mL) and (4-bromomethyl)benzoic acid methyl ester (6.60 g, 28.8 mmol) were added and the mixture was stirred at room temperature for 16 hours. Solvent was concentrated in vacuo and the resulting solid was partitioned between EtOAc and H 2 O. The organic layer was washed with brine and 1 N HCl, it was dried over MgSO 4 and concentrated under a vacuum. Using EtOAc / Hex (45:55) The crude material was purified by flash chromatography followed using MeOH / DCM (2:98) was purified by Biotage pre-packed silica gel column, and the resulting crystals 2 O in a mixture of CHCl 3 and Et are The title compound 152 (2.66 g, yield 19%). 1 H NMR: (DMsO-d 6 ) δ (ppm): 7.89 (d, J = 8.0 Hz, 2H), 7.87 (d, J = 7.8 Hz, 2H), 7.55 (d, J = 8.4 Hz, 2H) , 7.54 (d, J = 8.8 Hz, 1H), 7.47 (d, J = 8.4 Hz, 2H), 6.93 (d, J = 2.3 Hz, 1H), 6.77 (dd, J = 8.8, 2.5 Hz, 1H) , 6.70 (t, J = 6.1 Hz, 1H), 4.58 (s, 2H), 4.38 (d, J = 6.3 Hz, 2H), 3.81 (s, 3H), 3.81 (s, 3H). m/z: 479.4 (MH + ).
步驟2：N－(4－甲基苯甲酸)－苯幷噻唑－2－基硫基甲基}－苯甲酸(156)： 依照與流程1步驟4(實例1)對水解所述相同之程序用化合物152作為原料且使氫氧化鋰量加倍獲得標題化合物156(產率37%)，m/z：451.4(MH＋ )。 Step 2: N-(4-methylbenzoic acid)-benzoquinone-2-ylthiomethyl}-benzoic acid (156): according to the same procedure as described for the hydrolysis of step 1 of Example 1 (Example 1) The title compound 156 (yield 37%) m/z: 451.4 (MH + )
步驟3：N－(2－胺基－苯基)－4－{6－[N－(2－胺基－苯基)－4－苯甲醯胺]－苯幷噻唑－2－基硫基甲基}－苯甲醯胺(160) 依照與流程1步驟5(實例1)對BOP偶合所述相同之程序用化合物156作為原料且使所有試劑量加倍來獲得標題化合物160(產率5%)。1 H NMR：(丙酮－d6 )d(ppm)：7.98(d,J＝8.0 Hz,2H),7.96(d,J＝8.2 Hz,2H),7.63(d,J＝9.0 Hz,1H),7.62(d,J＝7.4 Hz,2H),7.55(d,J＝8.0 Hz,2H),7.28(d,J＝8.2 Hz,2H),7.04(d,J＝2.5 Hz,1H),6.99(t,J＝7.4 Hz,2H),6.91(dd,J＝8.8,2.3 Hz,1H),6.85(d,J＝7.4 Hz,2H),6.66(t,J＝7.4 Hz,2H),4.65(s,2H),4.54(s,2H)。m/z：631.5(MH＋ )。 Step 3: N-(2-Amino-phenyl)-4-{6-[N-(2-amino-phenyl)-4-benzylideneamine]-benzoquinone-2-ylthio Methyl}-benzamide (160) was subjected to the same procedure as described for the coupling of BOP in Step 1 of Example 1 (Example 1), using Compound 156 as a starting material and doubling the amount of all reagents to obtain the title compound 160 (yield 5%) ). 1 H NMR: (acetone-d 6 )d (ppm): 7.98 (d, J = 8.0 Hz, 2H), 7.96 (d, J = 8.2 Hz, 2H), 7.63 (d, J = 9.0 Hz, 1H) , 7.62 (d, J = 7.4 Hz, 2H), 7.55 (d, J = 8.0 Hz, 2H), 7.28 (d, J = 8.2 Hz, 2H), 7.04 (d, J = 2.5 Hz, 1H), 6.99 (t, J = 7.4 Hz, 2H), 6.91 (dd, J = 8.8, 2.3 Hz, 1H), 6.85 (d, J = 7.4 Hz, 2H), 6.66 (t, J = 7.4 Hz, 2H), 4.65 (s, 2H), 4.54 (s, 2H). m/z: 631.5 (MH + ).
N－(2－胺基－苯基)－4－(6－甲氧基－苯幷噻唑－2－基胺基)－苯甲醯胺(166)步驟1：4－(6－甲氧基－苯幷噻唑－2－基胺基)－苯甲酸甲酯(161)： 向2－氯－6－甲氧基苯幷噻唑(1.00 g，5.03 mmol)之DMF(10 mL)溶液中加入4－胺基苯甲酸甲酯(760 mg，5.03 mmol)隨後加入粉末狀K2 CO3(1.81 g，15.09 mmol)。將該混合物在90℃下攪拌16小時且在120℃下攪拌24小時且接著在140℃下攪拌3天。使其冷卻至室溫且添加NaH(在礦物油中60%，201 mg，5.03 mmol)。在室溫下攪拌混合物16小時且以H2 O將其中止。在80℃下真空移除溶劑且使殘餘物在H2 O與EtOAc之間分溶。以HCl 1 N、飽和NaHCO3 及鹽水洗滌有機層，使其經MgSO4 乾燥，且在真空下濃縮。由急驟層析法使用EtOAc/Hex且溶離期間將極性自20：80增加至50：50純化粗材料產生標題化合物161(150 mg，產率9%)，m/z：315.2(MH＋ )。 N-(2-Amino-phenyl)-4-(6-methoxy-benzothiazol-2-ylamino)-benzamide (166) Step 1: 4-(6-methoxy -Benzyl thiazol-2-ylamino)-benzoic acid methyl ester (161): To a solution of 2-chloro-6-methoxybenzothiazole (1.00 g, 5.03 mmol) in DMF (10 mL) - amino benzoic acid methyl ester (760 mg, 5.03 mmol) was added followed by powdered K 2 CO3 (1.81 g, 15.09 mmol). The mixture was stirred at 90 ° C for 16 hours and at 120 ° C for 24 hours and then at 140 ° C for 3 days. It was allowed to cool to room temperature and NaH (60% in mineral oil, 201 mg, 5.03 mmol). The mixture was stirred for 16 hours at room temperature and H 2 O in which to stop. The solvent was removed in vacuo <RTI ID=0.0>: </RTI><RTIgt; The organic layer HCl 1 N, saturated NaHCO 3 and brine, dried over MgSO 4, and concentrated in vacuo. During by flash chromatography using EtOAc / Hex and the polarity of the eluting from 20:80 to 50:50 increased crude material was purified to give the title compound 161 (150 mg, yield 9%), m / z: 315.2 (MH +).
步驟2：4－(6－甲氧基－苯幷噻唑－2－基胺基)－苯甲酸(163)： 依照與流程1步驟4(實例1)對水解所述相同之程序用化合物161作為原料獲得標題化合物163(產率66%)。m/z：301.2(MH＋ )。 Step 2: 4-(6-Methoxy-benzothiazol-2-ylamino)-benzoic acid (163): Compound 161 was used according to the same procedure as described for the hydrolysis of Step 1 of Example 1 (Example 1) The title compound 163 (yield: 66%) was obtained. m/z: 301.2 (MH + ).
步驟3：N－(2－胺基－苯基)－4－(6－甲氧基－苯幷噻唑－2－基胺基)－苯甲醯胺(166) 依照與流程1步驟5(實例1)對BOP偶合所述相同之程序用化合物163作為原料獲得標題化合物166(產率53%)。1 H NMR：(DMSO－d6 )δ(ppm)：10.62(s,1H),9.53(s,1H),7.98(d,J＝8.8 Hz,2H),7.84(d,J＝9.2 Hz,2H),7.57(d,J＝8.8 Hz,1H),7.47(d,J＝2.0 Hz,1H),6.59(t,J＝7.2 Hz,1H),4.89(s,2H),3.78(s,3H)。m/z：391.4(MH＋ )。 Step 3: N-(2-Amino-phenyl)-4-(6-methoxy-benzothiazol-2-ylamino)-benzamide (166) according to step 5 of Scheme 1 (example) 1) The title compound 166 (yield: 53%) was obtained. 1 H NMR: (DMSO-d 6 ) δ (ppm): 10.62 (s, 1H), 9.53 (s, 1H), 7.78 (d, J = 8.8 Hz, 2H), 7.84 (d, J = 9.2 Hz, 2H), 7.57 (d, J = 8.8 Hz, 1H), 7.47 (d, J = 2.0 Hz, 1H), 6.59 (t, J = 7.2 Hz, 1H), 4.89 (s, 2H), 3.78 (s, 3H). m/z: 391.4 (MH + ).
N－(2－胺基－苯基)－4－[3－(6－甲氧基－苯幷噻唑－2－基)－脲基]－苯甲醯胺(167)步驟1：4－[3－(6－甲氧基－苯幷噻唑－2－基)－脲基]－苯甲酸乙酯(162)： 依照J.Med.Chem.,1979,22(1),28－32所述之程序自2－胺基－6－甲氧基苯幷噻唑開始獲得標題化合物162(產率93%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.63(bs,1H),7.91(d,J＝8.4 Hz,2H),7.65(d,J＝8.0 Hz,2H),7.55－7.51(m,2H),6.98(d,J＝8.8 Hz,1H),4.28(q,J＝6.8 Hz,2H),3.79(s,3H),1.32(t,J＝7.2 Hz,3H)。m/z：372.3(MH＋ )。 N-(2-Amino-phenyl)-4-[3-(6-methoxy-benzothiazol-2-yl)-ureido]-benzamide (167) Step 1: 4-[ 3-(6-Methoxy-benzothiazol-2-yl)-ureido]-benzoic acid ethyl ester (162): according to J. Med. Chem., 1979, 22(1), 28-32. Procedure The title compound 162 (yield: 93%) was obtained from 2-amino-6-methoxybenzothiazole. 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.63 (bs, 1H), 7.91 (d, J = 8.4 Hz, 2H), 7.65 (d, J = 8.0 Hz, 2H), 7.55-7.51 ( m, 2H), 6.98 (d, J = 8.8 Hz, 1H), 4.28 (q, J = 6.8 Hz, 2H), 3.79 (s, 3H), 1.32 (t, J = 7.2 Hz, 3H). m/z: 372.3 (MH + ).
步驟2：4－[3－(6－甲氧基－苯幷噻唑－2－基)－脲基]－苯甲酸(164)： 依照與流程1步驟4(實例1)對水解所述相同之程序用化合物162作為原料獲得標題化合物164(產率99%)。1 H NMR：(DMSO－d6 )δ(ppm)：7.94(d,J＝8.4 Hz,2H),7.70(d,J＝8.0 Hz,2H),7.57(d,J＝8.4 Hz,1H),7.49(d,J＝2.4 Hz,1H),6.96(dd,J＝8.8,2.4 Hz,1H),3.80(s,3H)。m/z：344.3(MH＋ )。 Step 2: 4-[3-(6-Methoxy-benzothiazol-2-yl)-ureido]-benzoic acid (164): identical to that described for the hydrolysis of Step 4 (Example 1) of Scheme 1 Procedure The title compound 164 (yield: 99%) was obtained from Compound 162. 1 H NMR: (DMSO-d 6 ) δ (ppm): 7.94 (d, J = 8.4 Hz, 2H), 7.70 (d, J = 8.0 Hz, 2H), 7.57 (d, J = 8.4 Hz, 1H) , 7.49 (d, J = 2.4 Hz, 1H), 6.96 (dd, J = 8.8, 2.4 Hz, 1H), 3.80 (s, 3H). m/z: 344.3 (MH + ).
步驟3：N－(2－胺基－苯基)－4－[3－(6－甲氧基－苯幷噻唑－2－基)－脲基]－苯甲醯胺(167) 依照與流程1步驟5(實例1)對BOP偶合所述相同之程序用化合物164作為原料獲得標題化合物167(產率50%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.58(s,H),9.54(bs,1H),7.96(d,J＝8.4 Hz,2H),7.93(s,1H),7.63(d,J＝8.8 Hz,2H),7.54(d,J＝9.2 Hz,1H),7.52(d,J＝2.0 Hz,1H),7.14(d,J＝7.6 Hz,1H),6.98(dd,J＝8.0,1.6 Hz,1H),6.94(d,J＝8.0 Hz,1H),6.77(d,J＝8.0 Hz,1H),6.59(t,J＝7.2 Hz,1H),4.89(bs,2H),3.80(s,3H)。m/z：434.4(MH＋ )。 Step 3: N-(2-Amino-phenyl)-4-[3-(6-methoxy-benzothiazol-2-yl)-ureido]-benzamide (167) according to the procedure 1 Step 5 (Example 1) The title compound 167 (yield 50%) was obtained from the same procedure using the compound 164. 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.58 (s, H), 9.54 (bs, 1H), 7.96 (d, J = 8.4 Hz, 2H), 7.93 (s, 1H), 7.63 ( d, J = 8.8 Hz, 2H), 7.54 (d, J = 9.2 Hz, 1H), 7.52 (d, J = 2.0 Hz, 1H), 7.14 (d, J = 7.6 Hz, 1H), 6.98 (dd, J = 8.0, 1.6 Hz, 1H), 6.94 (d, J = 8.0 Hz, 1H), 6.77 (d, J = 8.0 Hz, 1H), 6.59 (t, J = 7.2 Hz, 1H), 4.89 (bs, 2H), 3.80 (s, 3H). m/z: 434.4 (MH + ).
N－(2－胺基－苯基)－4－[3－(6－甲氧基－苯幷噻唑－2－基)－脲甲基]－苯甲醯胺(168)步驟1：4－[3－(6－甲氧基－苯幷噻唑－2－基)－脲甲基]－苯甲酸(165)： 依照與流程32步驟1(實例75)對形成胺基甲酸酯所述相同之程序，以2－胺基－6－甲氧基苯幷噻唑代替化合物129且使用4－胺甲基苯甲酸而非4－(2－胺乙基)－嗎啉獲得標題化合物165(產率28%)。1 H NMR：(DMSO－d6 )δ(ppm)：7.92(t,J＝8.0 Hz,2H),7.51(d,J＝8.5 Hz,1H),7.47(s,1H),7.42(d,J＝8.5 Hz,1H),6.95(d,J＝7.0 Hz,1H),4.45(s,2H),3.77(s,3H)。m/z：358.3(MH＋ )。 N-(2-Amino-phenyl)-4-[3-(6-methoxy-benzothiazol-2-yl)-ureamethyl]-benzamide (168) Step 1: 4- [3-(6-Methoxy-benzothiazol-2-yl)-ureamethyl]-benzoic acid (165): identical to that described for the formation of the urethane with Step 32 (Example 75) of Scheme 32 Procedure, the title compound 165 was obtained by substituting 2-amino-6-methoxybenzothiazole for compound 129 and using 4-aminomethylbenzoic acid instead of 4-(2-aminoethyl)-morpholine. 28%). 1 H NMR: (DMSO-d 6 ) δ (ppm): 7.92 (t, J = 8.0 Hz, 2H), 7.51 (d, J = 8.5 Hz, 1H), 7.47 (s, 1H), 7.42 (d, J = 8.5 Hz, 1H), 6.95 (d, J = 7.0 Hz, 1H), 4.45 (s, 2H), 3.77 (s, 3H). m/z: 358.3 (MH + ).
步驟2：N－(2－胺基－苯基)－4－[3－(6－甲氧基－苯幷噻唑－2－基)－脲甲基]－苯甲醯胺(168) 依照與流程1步驟5(實例1)對BOP偶合所述相同之程序用化合物165作為原料獲得標題化合物168(產率1.5%)。1 H NMR：(DMSO－d6 )δ(ppm)：10.75(bs,14H),9.63(s,1H),7.97－7.91(m,2H),7.53－7.43(m,3H),7.33(s,1H),7.16(s,1H),6.96－6.95(m,2H),6.78(d,J＝8.0 Hz,1H),6.60－6.58(m,1H),4.88(bs,2H),4.45(s,2H),3.78(s,3H)。 Step 2: N-(2-Amino-phenyl)-4-[3-(6-methoxy-benzothiazol-2-yl)-ureamethyl]-benzamide (168) according to Scheme 1, Step 5 (Example 1) The title compound 168 (yield: 1.5%). 1 H NMR: (DMSO-d 6 ) δ (ppm): 10.75 (bs, 14H), 9.63 (s, 1H), 7.97-7.91 (m, 2H), 7.53-7.43 (m, 3H), 7.33 (s) , 1H), 7.16 (s, 1H), 6.96-6.95 (m, 2H), 6.78 (d, J = 8.0 Hz, 1H), 6.60-6.58 (m, 1H), 4.88 (bs, 2H), 4.45 ( s, 2H), 3.78 (s, 3H).
N－(2－胺基－苯基)－4－{[5－(吡啶－2－基硫基)－噻唑－2－基胺基]－甲基}－苯甲醯胺(177)步驟1：5－(吡啶－2－基硫基)－噻唑－2－基胺(169)： 向氫溴化2－胺基－5－溴噻唑(1.00 g，3.85 mmol)之DMF(8 mL)溶液中加入2－巰基吡啶(428 mg，3.85 mmol)隨後加入粉末狀K2 CO3 (1.81 g，15.09 mmol)。將混合物在80℃下攪拌1小時且在室溫下攪拌16小時。在80℃下真空移除溶劑且使化合物在H2 O與EtOAc之間分溶。以EtOAc萃取含水層且以HCl 1 N萃取有機層。以飽和NaHCO3 中和酸性萃取物且以EtOAc萃取沉澱物，將該沉澱物以鹽水洗滌，經MgSO4 乾燥且在真空下濃縮產生標題化合物169(589 mg，產率73%)。1 H NMR：(丙酮－d6 )δ(ppm)：8.36(s,1H),7.66(s,1H),7.20(s,1H),7.12－7.05(m,2H),6.84(s,2H)。m/z：210.1(MH＋ )。 N-(2-Amino-phenyl)-4-{[5-(pyridin-2-ylthio)-thiazol-2-ylamino]-methyl}-benzamide (177) Step 1 :5-(Pyridin-2-ylthio)-thiazol-2-ylamine (169): To a solution of 2-amino-5-bromothiazole (1.00 g, 3.85 mmol) in DMF (8 mL) was added 2-mercaptopyridine (428 mg, 3.85 mmol) was added followed by powdered K 2 CO 3 (1.81 g, 15.09 mmol). The mixture was stirred at 80 ° C for 1 hour and at room temperature for 16 hours. The solvent was removed in vacuo at 80 deg.] C, and the compound was partitioned between H 2 O and EtOAc. The aqueous layer was extracted with EtOAc and EtOAc EtOAc. Saturated NaHCO 3 to neutralize the acidic and extracted with EtOAc extract the precipitate, the precipitate was washed with brine, dried over MgSO 4 and concentrated under vacuo to give the title compound 169 (589 mg, 73% yield). 1 H NMR: (acetone-d 6 ) δ (ppm): 8.36 (s, 1H), 7.66 (s, 1H), 7.20 (s, 1H), 7.12-7.05 (m, 2H), 6.84 (s, 2H) ). m/z: 210.1 (MH + ).
步驟2：4－{[5－(吡啶－2－基硫基)－噻唑－2－基胺基]－甲基}－苯甲酸甲酯(173)： 依照與流程3步驟2(實例12)對還原胺化反應所述相同之程序自化合物169開始獲得標題化合物173(產率50%)。1 H NMR：(丙酮－d6 )δ(ppm)：8.37(d,J＝4.0 Hz,1H),7.99(d,J＝8.5 Hz,2H),7.83(bs,1H),7.67(td,J＝8.0,1.5 Hz,1H),7.56(d,J＝7.5 Hz,2H),7.28(s,1H),7.13(dd,J＝6.5,5.0 Hz,1H),7.07(d,J＝8.0 Hz,1H),4.72(bs,2H),3.88(s,3H)。m/z：358.1(MH＋ )。 Step 2: 4-{[5-(Pyridin-2-ylthio)-thiazol-2-ylamino]-methyl}-benzoic acid methyl ester (173): according to step 2 of Scheme 3 (Example 12) The same procedure as described for the reductive amination reaction afforded the title compound 173 (yield 50%) starting from compound 169. 1 H NMR: (acetone-d 6 ) δ (ppm): 8.37 (d, J = 4.0 Hz, 1H), 7.99 (d, J = 8.5 Hz, 2H), 7.83 (bs, 1H), 7.67 (td, J = 8.0, 1.5 Hz, 1H), 7.56 (d, J = 7.5 Hz, 2H), 7.28 (s, 1H), 7.13 (dd, J = 6.5, 5.0 Hz, 1H), 7.07 (d, J = 8.0) Hz, 1H), 4.72 (bs, 2H), 3.88 (s, 3H). m/z: 358.1 (MH + ).
步驟3：4－{[5－(吡啶－2－基硫基)－噻唑－2－基胺基]－甲基}－苯甲酸(175)： 依照與流程1步驟4(實例1)對水解所述相同之程序用化合物173作為原料獲得標題化合物175(產率81%)。1 H NMR：(丙酮－d6 )δ(ppm)：8.37(d,J＝4.0 Hz,1H),7.99(d,J＝8.5 Hz,2H),7.83(bs,1H),7.67(td,J＝8.0,1.5 Hz,1H),7.56(d,J＝7.5 Hz,2H),7.28(s,1H),7.13(dd,J＝6.5,5.0 Hz,1H),7.07(d,J＝8.0 Hz,1H),4.72(bs,2H),3.88(s,3H)。m/z：344.0(MH＋ )。 Step 3: 4-{[5-(Pyridin-2-ylthio)-thiazol-2-ylamino]-methyl}-benzoic acid (175): hydrolyzed according to Step 4 (Example 1) of Scheme 1 The same procedure was used to obtain the title compound 175 (yield: 81%). 1 H NMR: (acetone-d 6 ) δ (ppm): 8.37 (d, J = 4.0 Hz, 1H), 7.99 (d, J = 8.5 Hz, 2H), 7.83 (bs, 1H), 7.67 (td, J = 8.0, 1.5 Hz, 1H), 7.56 (d, J = 7.5 Hz, 2H), 7.28 (s, 1H), 7.13 (dd, J = 6.5, 5.0 Hz, 1H), 7.07 (d, J = 8.0) Hz, 1H), 4.72 (bs, 2H), 3.88 (s, 3H). m/z: 344.0 (MH + ).
步驟4：N－(2－胺基－苯基)－4－{[5－(吡啶－2－基硫基)－噻唑－2－基胺基]－甲基}－苯甲醯胺(177) 依照與流程1步驟5(實例1)對BOP偶合所述相同之程序用化合物175作為原料獲得標題化合物177(產率53%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.63(s,1H),8.74(t,J＝5.9 Hz,1H),8.40(d,J＝3.7 Hz,1H),7.96(d,J＝8.4 Hz,2H),7.72(td,J＝7.6,2.0 Hz,1H),7.48(d,J＝7.8 Hz,2H),7.34(s,1H),7.19－7.15(m,2H),7.05(d,J＝8.2 Hz,1H),6.97(t,J＝8.0 Hz,1H),6.78(d,J＝7.8 Hz,1H),6.60(t,J＝7.8 Hz,1H),4.91(s,2H),4.59(d,J＝6.1 Hz,2H)。mz：434.4(MH＋ )。 Step 4: N-(2-Amino-phenyl)-4-{[5-(pyridin-2-ylthio)-thiazol-2-ylamino]-methyl}-benzamide (177 The title compound 177 (yield: 53%) was obtained from compound 175 (yield: mp. 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.63 (s, 1H), 8.74 (t, J = 5.9 Hz, 1H), 8.40 (d, J = 3.7 Hz, 1H), 7.96 (d, J = 8.4 Hz, 2H), 7.72 (td, J = 7.6, 2.0 Hz, 1H), 7.48 (d, J = 7.8 Hz, 2H), 7.34 (s, 1H), 7.19-7.15 (m, 2H), 7.05 (d, J = 8.2 Hz, 1H), 6.97 (t, J = 8.0 Hz, 1H), 6.78 (d, J = 7.8 Hz, 1H), 6.60 (t, J = 7.8 Hz, 1H), 4.91 ( s, 2H), 4.59 (d, J = 6.1 Hz, 2H). Mz: 434.4 (MH + ).
N－(2－胺基－苯基)－4－{[5－(吡啶－2－基氧基)－噻唑－2－基胺基]－甲基}－苯甲醯胺(178)步驟1：5－(吡啶－3－基氧基)－噻唑－2－基胺(170)： 向NaH(在礦物油中60%，169 mg，4.23 mmol)之DME(10 mL)懸浮液中加入2－羥基吡啶(366 mg，3.85 mmol)。[觀察到氫析出]。接著添加粉末狀K2 CO3 (2.31g，19.2 mmol)隨後逐份加入氫溴化2－胺基－5－溴噻唑(1.00 g，3.85 mmol)。將混合物在攪拌下回流16小時且使其冷卻至室溫，以水中止且使其在水與EtOAc之間分溶。以EtOAc萃取含水層且以HCl 1 N萃取有機相。以飽和NaHCO3 中和酸性萃取物，且先以EtOAc接著以MeOH/CHCl3 (20：85)之混合物萃取該沉澱物。使合併之有機萃取物經MgSO4 乾燥且在真空下濃縮。藉由加入MeOH/CHCl3 (5：95)之混合物使殘餘物結晶產生標題化合物170(21 mg，3%)。1 H NMR：(CD3 OD)δ(ppm)：9.05(dd,J＝7.2,2.0 Hz,1H),8.70(ddd,J＝9.2,6.7,2.2 Hz,1H),8.44(s,1H),7.73(d,J＝8.8 Hz,1H),7.59(bs,2H),7.58(td,J＝6.8,1.4 Hz,1H)。m/z：194.2(MH＋ )。 N-(2-Amino-phenyl)-4-{[5-(pyridin-2-yloxy)-thiazol-2-ylamino]-methyl}-benzamide (178) Step 1 :5-(Pyridin-3-yloxy)-thiazol-2-ylamine (170): To a suspension of NaH (60% in mineral oil, 169 mg, 4.23 mmol) in DME (10 mL) -Hydroxypyridine (366 mg, 3.85 mmol). [Hydrogen precipitation was observed]. Powdered K 2 CO 3 (2.31 g, 19.2 mmol) was then added followed by 2-amino-5-bromothiazole hydrobromide (1.00 g, 3.85 mmol). The mixture was refluxed for 16 hours with stirring and allowed to cool to room temperature, then quenched with water and partitioned between water and EtOAc. The aqueous layer was extracted with EtOAc and EtOAc was evaporated. Saturated NaHCO 3 to neutralize the acidic extracts were first with EtOAc and then a mixture of MeOH / CHCl 3 (20:85) extracts of the precipitate. The combined organic extracts were dried with MgSO 4 By addition of MeOH / CHCl 3 mixture (5:95) of the residue was crystallized to give the title compound 170 (21 mg, 3%) . 1 H NMR: (CD 3 OD) δ (ppm): 9.05 (dd, J = 7.2, 2.0 Hz, 1H), 8.70 (ddd, J = 9.2, 6.7, 2.2 Hz, 1H), 8.44 (s, 1H) , 7.73 (d, J = 8.8 Hz, 1H), 7.59 (bs, 2H), 7.58 (td, J = 6.8, 1.4 Hz, 1H). m/z: 194.2 (MH + ).
步驟2：[2－(4－{[5－(吡啶－3－基氧基)－噻唑－2－基胺基]－甲基}－苯甲醯胺基)－苯基]－胺基甲酸第三丁酯(173)： 依照與流程3步驟2(實例12)對還原胺化反應所述相同之程序使化合物170與化合物128反應(描述於專利申請案WO 03/024448中)獲得標題化合物173(產率46%)。1 H NMR：(丙酮－d6 )δ(ppm)：9.66(s,1H),8.30(s,1H),7.97(d,J＝8.2 Hz,2H),7.79(ddd,J＝7.0,2.0,0.6 Hz,1H),7.68(dd,J＝7.6,1.6 Hz,1H),7.60(dd,J＝7.8,1.8 Hz,1H),7.55(d,J＝8.2 Hz,2H),7.45(ddd,J＝9.4,6.7,2.0 Hz,1H),7.27(s,1H),7.21(td,J＝7.4,1.8 Hz,1H),7.16(dt,J＝7.4,1.8 Hz,1H),6.50(d,J＝9.2 Hz,1H),6.33(td,J＝6.7,1.4 Hz,1H),4.67(s,2H),1.99(s,9H)。m/z：518.5(MH＋ )。 Step 2: [2-(4-{[5-(Pyridin-3-yloxy)-thiazol-2-ylamino]-methyl}-benzylidene)-phenyl]-aminocarboxylic acid Third butyl ester (173): The title compound was obtained by reacting compound 170 with compound 128 (described in patent application WO 03/024448) following the same procedure as described for the reductive amination reaction of Scheme 3, Step 2 (Example 12). 173 (yield 46%). 1 H NMR: (acetone-d 6 ) δ (ppm): 9.66 (s, 1H), 8.30 (s, 1H), 7.97 (d, J = 8.2 Hz, 2H), 7.79 (ddd, J = 7.0, 2.0) , 0.6 Hz, 1H), 7.68 (dd, J = 7.6, 1.6 Hz, 1H), 7.60 (dd, J = 7.8, 1.8 Hz, 1H), 7.55 (d, J = 8.2 Hz, 2H), 7.45 (ddd , J = 9.4, 6.7, 2.0 Hz, 1H), 7.27 (s, 1H), 7.21 (td, J = 7.4, 1.8 Hz, 1H), 7.16 (dt, J = 7.4, 1.8 Hz, 1H), 6.50 ( d, J = 9.2 Hz, 1H), 6.33 (td, J = 6.7, 1.4 Hz, 1H), 4.67 (s, 2H), 1.99 (s, 9H). m/z: 518.5 (MH + ).
步驟3：N－(2－胺基－苯基)－4－{[5－(吡啶－2－基氧基)－噻唑－2－基胺基]－甲基}－苯甲醯胺(178) 依照與流程28步驟5(實例68)對Boc***所述相同之程序以化合物171作為原料獲得標題化合物178(產率82%)。1 H NMR：(丙酮－d6 )δ(ppm)：8.00(d,J＝8.4 Hz,2H),7.82(dd,J＝6.3,1.4 Hz,1H),7.56(d,J＝8.2 Hz,2H),7.46(ddd,J＝13.7,6.7,2.2 Hz,1H),7.30(d,J＝6.7 Hz,1H),7.28(s,1H),6.99(td,J＝13.7,7.2 Hz,1H),6.87(dd,J＝6.7,1.2 Hz,1H),6.67(t,J＝7.2 Hz,1H),6.49(d,J＝8.8 Hz,1H),6.34(td,J＝6.7,5.3 Hz,1H),4.69(s,2H)。m/z：434.4(MH＋ )。 Step 3: N-(2-Amino-phenyl)-4-{[5-(pyridin-2-yloxy)-thiazol-2-ylamino]-methyl}-benzamide (178 The title compound 178 (yield: 82%) was obtained from compound 171 (yield). 1 H NMR: (acetone-d 6 ) δ (ppm): 8.00 (d, J = 8.4 Hz, 2H), 7.82 (dd, J = 6.3, 1.4 Hz, 1H), 7.56 (d, J = 8.2 Hz, 2H), 7.46 (ddd, J = 13.7, 6.7, 2.2 Hz, 1H), 7.30 (d, J = 6.7 Hz, 1H), 7.28 (s, 1H), 6.99 (td, J = 13.7, 7.2 Hz, 1H) ), 6.87 (dd, J = 6.7, 1.2 Hz, 1H), 6.67 (t, J = 7.2 Hz, 1H), 6.49 (d, J = 8.8 Hz, 1H), 6.34 (td, J = 6.7, 5.3 Hz) , 1H), 4.69 (s, 2H). m/z: 434.4 (MH + ).
N－(2－胺基－苯基)－4－[(4－吡啶－3－基－噻唑－2－基胺基)－甲基]－苯甲醯胺(179)步驟1：4－吡啶－3－基－噻唑－2－基胺(172)： 依照J.Heterocycl.Chem.,1970,7,(1137－1141)所述之程序獲得標題化合物172(產率94%)。1 H NMR：(CD3 OD)δ(ppm)：8.94(dd,J＝2.3,0.8 Hz,1H),8.41(dd,J＝4.7,1.6 Hz,1H),8.18(dt,J＝8.6,1.6 Hz,1H),7.43(ddd,J＝9.0,3.9,0.8 Hz,1H),7.03(s,1H)。m/z：178.1(MH＋ )。 N-(2-Amino-phenyl)-4-[(4-pyridin-3-yl-thiazol-2-ylamino)-methyl]-benzamide (179) Step 1: 4-pyridine 3-yl-thiazol-2-ylamine (172): The title compound 172 (yield: 94%) was obtained according to the procedure of J. Heterocycl. Chem., 1970, 7, (1137-1141). 1 H NMR: (CD 3 OD) δ (ppm): 8.94 (dd, J = 2.3, 0.8 Hz, 1H), 8.41 (dd, J = 4.7, 1.6 Hz, 1H), 8.18 (dt, J = 8.6, 1.6 Hz, 1H), 7.43 (ddd, J = 9.0, 3.9, 0.8 Hz, 1H), 7.03 (s, 1H). m/z: 178.1 (MH + ).
步驟2：4－[(4－吡啶－3－基－噻唑－2－基胺基)－甲基]－苯甲酸甲酯(174)： 依照與流程3步驟2(實例12)對還原胺化反應所述相同之程序用化合物172作為原料獲得標題化合物174(產率33%)。1 H NMR：(丙酮－d6 )δ(ppm)：9.07(dd,J＝2.3,0.8 Hz,1H),8.45(dd,J＝4.7,1.6 Hz,1H),8.16(dt,J＝8.6,1.6 Hz,1H),7.98(d,J＝8.6 Hz,2H),7.60(d,J＝8.6 Hz,2H),7.52－7.49(m,1H),7.34(ddd,J＝7.8,4.7,0.8,1H),7.14(s,1H),4.76(s,2H),3.87(s,3H)。m/z：326.3(MH＋ )。 Step 2: 4-[(4-Pyridin-3-yl-thiazol-2-ylamino)-methyl]-benzoic acid methyl ester (174): reductive amination according to procedure 2, step 2 (Example 12) The title compound 174 (yield 33%) was obtained from the title compound 172. 1 H NMR: (acetone-d 6 ) δ (ppm): 9.07 (dd, J = 2.3, 0.8 Hz, 1H), 8.45 (dd, J = 4.7, 1.6 Hz, 1H), 8.16 (dt, J = 8.6 , 1.6 Hz, 1H), 7.98 (d, J = 8.6 Hz, 2H), 7.60 (d, J = 8.6 Hz, 2H), 7.52-7.49 (m, 1H), 7.34 (ddd, J = 7.8, 4.7, 0.8, 1H), 7.14 (s, 1H), 4.76 (s, 2H), 3.87 (s, 3H). m/z: 326.3 (MH + ).
步驟3：4－[(4－吡啶－3－基－噻唑－2－基胺基)－甲基]－苯甲酸(176)： 依照與流程1步驟4(實例1)對水解所述相同之程序用化合物174作為原料獲得標題化合物176(產率27%)。1 H NMR：(DMSO－d6 )δ(ppm)：8.99(dd,J＝2.0,0.8,1H),8.42(dd,J＝4.7,1.6 Hz,1H),8.23(t,J＝5.9 Hz,1H),8.11(dt,J＝8.2,2.0 Hz,1H),7.76(d,J＝8.2 Hz,2H),7.36(ddd,J＝7.8,4.7,0.8 Hz,1H),7.23(d,J＝8.2 Hz,2H),7.21(s,1H),7.47(d,J＝5.5 Hz,2H)。m/z：312.3(MH＋ )。 Step 3: 4-[(4-Pyridin-3-yl-thiazol-2-ylamino)-methyl]-benzoic acid (176): identical to that described for the hydrolysis of Step 4 (Example 1) of Scheme 1 Procedure The title compound 176 (yield 27%) was obtained from compound 174. 1 H NMR: (DMSO-d 6 ) δ (ppm): 8.99 (dd, J = 2.0, 0.8, 1H), 8.42 (dd, J = 4.7, 1.6 Hz, 1H), 8.23 (t, J = 5.9 Hz) , 1H), 8.11 (dt, J = 8.2, 2.0 Hz, 1H), 7.76 (d, J = 8.2 Hz, 2H), 7.36 (ddd, J = 7.8, 4.7, 0.8 Hz, 1H), 7.23 (d, J = 8.2 Hz, 2H), 7.21 (s, 1H), 7.47 (d, J = 5.5 Hz, 2H). m/z: 312.3 (MH + ).
步驟4：N－(2－胺基－苯基)－4－[(4－吡啶－3－基－噻唑－2－基胺基)－甲基]－苯甲醯胺(179) 依照與流程1步驟5(實例1)對BOP偶合反應所述相同之程序用化合物176作為原料獲得標題化合物179(產率94%)。1 H NMR(CDCl3 )δ(ppm)：10.00(s,1H),9.00(dd,J＝3.1,0.8 Hz,1H),8.43(dd,J＝4.7,1.6 Hz,1H),8.33(t,J＝6.3 Hz,1H),8.11(dt,J＝7.8,2.3 Hz,1H),7.89(d,J＝8.6 Hz,2H),7.62(dd,J＝5.9,3.5 Hz,1H),7.50(d,J＝8.6 Hz,2H),7.37(dd,J＝7.8,4.7 Hz,1H),7.26(dd,J＝5.5,3.5 Hz,1H),7.24(s,1H),4.56(d,J＝5.9 Hz,2H)。m/z：402.1(MH＋ )。 Step 4: N-(2-Amino-phenyl)-4-[(4-pyridin-3-yl-thiazol-2-ylamino)-methyl]-benzamide (179) according to the procedure 1 Step 5 (Example 1) For the BOP coupling reaction, the same procedure was used to obtain the title compound 179 (yield: 94%). 1 H NMR (CDCl 3 ) δ (ppm): 10.00 (s, 1H), 9.00 (dd, J = 3.1, 0.8 Hz, 1H), 8.43 (dd, J = 4.7, 1.6 Hz, 1H), 8.33 (t) , J = 6.3 Hz, 1H), 8.11 (dt, J = 7.8, 2.3 Hz, 1H), 7.89 (d, J = 8.6 Hz, 2H), 7.62 (dd, J = 5.9, 3.5 Hz, 1H), 7.50 (d, J = 8.6 Hz, 2H), 7.37 (dd, J = 7.8, 4.7 Hz, 1H), 7.26 (dd, J = 5.5, 3.5 Hz, 1H), 7.24 (s, 1H), 4.56 (d, J = 5.9 Hz, 2H). m/z: 402.1 (MH + ).
N－(2－胺基－苯基)－4－[6－(2－二甲胺基－乙氧基)－5－氟－1H－苯幷咪唑－2－基硫基甲基]－苯甲醯胺(192)步驟1：5－(2－二甲胺基－乙氧基)－4－氟－2－硝基－苯胺(180)： 向一經烘乾(flame－dried)之圓底燒瓶中注入4,5－二氟－2－硝基苯胺(2.00 g，11.49 mmol)及N,N－二甲基乙醇胺。添加吡啶(44 mL)隨後緩緩添加NaH(礦物油中60%，965 mg，24.1 mmol)。將混合物置於N2 氣氛下，在室溫下攪拌16小時且以H2 O中止之。真空移除溶劑且使殘餘物在H2 O與EtOAc之間分溶。以HCl 1 N萃取有機層兩次，以飽和NaHCO3 中和合併之酸性萃取物形成沉澱物，使該沉澱物靜置隔夜，經過濾收集且由急驟層析法使用MeOH/CHCl3 同時增加極性(10：90至15：85)來純化該沉澱物產生標題化合物180(1.30 g，產率47%)。1 H NMR：(CD3 OD)δ(ppm)：7.76(d,J＝11.7 Hz,1H),6.53(d,J＝7.4 Hz,1H),4.19(t,J＝5.5 Hz,2H),2.84(t,J＝5.5 Hz,2H),2.37(s,6H)。m/z：244.2(MH＋ )。 N-(2-Amino-phenyl)-4-[6-(2-dimethylamino-ethoxy)-5-fluoro-1H-benzoimidazol-2-ylthiomethyl]-benzene Methionamine (192) Step 1: 5-(2-Dimethylamino-ethoxy)-4-fluoro-2-nitro-phenylamine (180): to a flame-dried round bottom The flask was charged with 4,5-difluoro-2-nitroaniline (2.00 g, 11.49 mmol) and N,N-dimethylethanolamine. Pyridine (44 mL) was added followed by the slow addition of NaH (60% in mineral oil, 965 mg, 24.1 mmol). The mixture was placed under N 2 atmosphere, stirred for 16 hours at room temperature and H 2 O in the suspension. The solvent was removed in vacuo and the residue was partitioned between H 2 O and EtOAc. The organic layer was extracted twice with HCl 1 N, saturated NaHCO 3 to neutralize the acidic extracts were combined to form a precipitate, the precipitate was allowed to stand overnight so, collected by filtration and purified by flash chromatography using MeOH / CHCl 3 while increasing polarity (10:90 to 15:85) to purify the precipitate to give the title compound 180 (1.30 g, yield 47%). 1 H NMR: (CD 3 OD) δ (ppm): 7.76 (d, J = 11.7 Hz, 1H), 6.53 (d, J = 7.4 Hz, 1H), 4.19 (t, J = 5.5 Hz, 2H), 2.84 (t, J = 5.5 Hz, 2H), 2.37 (s, 6H). m/z: 244.2 (MH + ).
步驟2：4－(2－二甲胺基－乙氧基)－5－氟－苯－1,2－二胺(181)： 將中間體180(220 mg，0.904 mmol)之乙酸(3.6 mL)溶液脫氣且置於N2 氣氛下。加入催化量之Pd(OH)2 且在室溫下氫化(1 atm)該黑色混合物16小時，將其經由矽藻土墊過濾且以MeOH沖洗之。在80℃下真空濃縮濾液產生與AcONHEt3 形成混合物之標題化合物181(252 mg，75%)。1 H NMR：(CD3 OD)δ(ppm)：6.54(d,J＝7.8 Hz,1H),6.51(d,J＝12.3 Hz,1H),4.21(t,J＝5.1 Hz,2H),3.40(t,J＝5.1 Hz,2H),2.89(s,6H)。m/z：214.1(MH＋ )。 Step 2: 4-(2-Dimethylamino-ethoxy)-5-fluoro-benzene-1,2-diamine (181): Intermediate 180 (220 mg, 0.904 mmol). The solution was degassed and placed under a N 2 atmosphere. A catalytic amount of Pd(OH) 2 was added and the black mixture was hydrogenated (1 atm) at room temperature for 16 hours, which was filtered through a pad of celite and rinsed with MeOH. At 80 deg.] C and the filtrate was concentrated to give the title AcONHEt 3 is formed of a mixture of compound 181 (252 mg, 75%) . 1 H NMR: (CD 3 OD) δ (ppm): 6.54 (d, J = 7.8 Hz, 1H), 6.51 (d, J = 12.3 Hz, 1H), 4.21. (t, J = 5.1 Hz, 2H), 3.40 (t, J = 5.1 Hz, 2H), 2.89 (s, 6H). m/z: 214.1 (MH + ).
步驟3：6－(2－二甲胺基－乙氧基)－5－氟－1H－苯幷咪唑－2－硫醇(184)： 依照J.Med.Chem.,1998,63,977－983所述之程序自化合物181開始獲得標題化合物184(產率96%)。1 H NMR：(CD3 OD)δ(ppm)：7.16(d,J＝1.2 Hz,0.5H),7.07(d,J＝10.4 Hz,1H),7.04(d,J＝7.2 Hz,0.5H),4.37(t,J＝4.9 Hz,2H),3.50(t,J＝5.1 Hz,2H),2.92(s,6H)。m/z：256.2(MH＋ )。 Step 3: 6-(2-Dimethylamino-ethoxy)-5-fluoro-1H-benzimidazole-2-thiol (184): according to J. Med. Chem., 1998, 63, 977-983. The title compound 184 (yield 96%) was obtained from compound 181. 1 H NMR: (CD 3 OD) δ (ppm): 7.16 (d, J = 1.2 Hz, 0.5H), 7.07 (d, J = 10.4 Hz, 1H), 7.04 (d, J = 7.2 Hz, 0.5H) ), 4.37 (t, J = 4.9 Hz, 2H), 3.50 (t, J = 5.1 Hz, 2H), 2.92 (s, 6H). m/z: 256.2 (MH + ).
步驟4：4－[6－(2－二甲胺－乙氧基)－5－氟－1H－苯幷咪唑－2－基硫基甲基]－苯甲酸(188)： 依照與流程27步驟1(實例66及67)對烷基化反應所述相同之程序使化合物184與4－溴－甲苯甲酸反應獲得標題化合物188(產率100%)。1 H NMR：(DMSO－d6 )δ(ppm)：12.65(s,1H),7.92(s,1H),7.83(d,J＝8.2 Hz,2H),7.51(d,J＝8.0 Hz,2H),7.30－7.27(m,1H),4.58(s,2H),4.40(t,J＝4.9 Hz,2H),3.54(t,J＝4.9 Hz,2H),2.88(s,6H)。m/z：390.2(MH＋ )。 Step 4: 4- [6- (2-dimethylamine-ethoxy) - phenyl Bing -1H- imidazol-5-fluoro-2-methyl thio] - benzoic acid (188): In accordance with process step 27 1 (Examples 66 and 67) The same procedure as described for the alkylation reaction. Compound 184 was reacted with 4-bromo-toluic acid to afford the title compound 188 (yield 100%). 1 H NMR: (DMSO-d 6 ) δ (ppm): 12.65 (s, 1H), 7.92 (s, 1H), 7.83 (d, J = 8.2 Hz, 2H), 7.51 (d, J = 8.0 Hz, 2H), 7.30-7.27 (m, 1H), 4.58 (s, 2H), 4.40 (t, J = 4.9 Hz, 2H), 3.54 (t, J = 4.9 Hz, 2H), 2.88 (s, 6H). m/z: 390.2 (MH + ).
步驟5：N－(2－胺基－苯基)－4－[6－(2－二甲胺基－乙氧基)－5－氟－1H－苯幷咪唑－2－基硫基甲基]－苯甲醯胺(192) 依照與流程1步驟5(實例1)對BOP偶合反應所述相同之程序用化合物188作為原料獲得標題化合物192(產率30%)。1 H NMR：(丙酮－d6 )δ(ppm)：9.02(bs,1H),7.95(d,J＝8.0 Hz,2H),7.61(d,J＝8.0 Hz,2H),7.27(d,J＝7.6 Hz,1H),7.28－7.10(m,2H),6.99(td,J＝8.0,1.6 Hz,1H),6.86(dd,J＝7.8,1.2 Hz,1H),6.66(t,J＝8.8 Hz,1H),4.65(s,2H),4.63(bs,2H),4.22(bs,2H),2.87(bs,2H),2.41(s,6H)。m/z：480.4(MH＋ )。 Step 5: N-(2-Amino-phenyl)-4-[6-(2-dimethylamino-ethoxy)-5-fluoro-1H-benzoimidazol-2-ylthiomethyl --Benzalamine (192) The title compound 192 (yield 30%) was obtained from the compound 188 as the starting material. 1 H NMR: (acetone-d 6 ) δ (ppm): 9.02 (bs, 1H), 7.95 (d, J = 8.0 Hz, 2H), 7.61 (d, J = 8.0 Hz, 2H), 7.27 (d, J = 7.6 Hz, 1H), 7.28-7.10 (m, 2H), 6.99 (td, J = 8.0, 1.6 Hz, 1H), 6.86 (dd, J = 7.8, 1.2 Hz, 1H), 6.66 (t, J = 8.8 Hz, 1H), 4.65 (s, 2H), 4.63 (bs, 2H), 4.22 (bs, 2H), 2.87 (bs, 2H), 2.41 (s, 6H). m / z: 480.4 (MH + ).
N－(2－胺基－苯基)－4－(5,6－二甲氧基－1H－苯幷咪唑－2－基硫基甲基)－苯甲醯胺(193)步驟1：4,5－二甲氧基－苯－1,2－二胺(182)： 將1,2－二甲氧基－4,5－二硝基苯(500 mg，2.19 mmol)之MeOH(10 mL)溶液脫氣且置於N2 氣氛下。將催化量之Pd/木炭(10%)以MeOH(1 mL)中止且作為MeOH懸浮液一次性轉移至該溶液中。添加乙酸(1.5 mL)且將該黑色混合物置於H2 氣氛下(1 atm)，在室溫下攪拌16小時。經由矽藻土墊過濾混合物且以MeOH沖洗之。在80℃下真空下濃縮濾液產生標題化合物182(無法自產物移除殘留之乙酸)。1 H NMR：(DMSO－d6 )δ(ppm)：6.23(s,2H),3.56(s,6H)。m/z：169.3.(MH＋ )。 N-(2-Amino-phenyl)-4-(5,6-dimethoxy-1H-benzoimidazol-2-ylthiomethyl)-benzamide (193) Step 1:4 ,5-dimethoxy-benzene-1,2-diamine (182): 1,2-dimethoxy-4,5-dinitrobenzene (500 mg, 2.19 mmol) in MeOH (10 mL) The solution was degassed and placed under a N 2 atmosphere. A catalytic amount of Pd/charcoal (10%) was quenched with MeOH (1 mL) and transferred to the solution as a MeOH suspension. Acetic acid (1.5 mL) was added and the black mixture was placed under H 2 atmosphere (1 atm) and stirred at room temperature for 16 hours. The mixture was filtered through a pad of celite and rinsed with MeOH. Concentration of the filtrate under vacuum at <RTI ID=0.0></RTI><RTIID=0.0> 1 H NMR: (DMSO-d 6 ) δ (ppm): 6.23 (s, 2H), 3.56 (s, 6H). m/z: 169.3. (MH + ).
步驟2：5,6－二甲氧基－1H－苯幷咪唑－2－硫醇(185)： 依照J.Med.Chem.,1998,63,977－983所述之程序自化合物182開始獲得標題化合物185(經兩個步驟產率44%)。1 H NMR：(DMSO－d6 )δ(ppm)：12.29(s,2H),6.71(s,2H),3.74(s,6H)。m/z：211.2(MH＋ )。 Step 2: 5,6-Dimethoxy-1H-benzimidazole-2-thiol (185): title compound was obtained from compound 182 according to the procedure described in J. Med. Chem., 1998, 63, 977-983. 185 (44% yield by two steps). 1 H NMR: (DMSO-d 6) δ (ppm): 12.29 (s, 2H), 6.71 (s, 2H), 3.74 (s, 6H). m/z: 211.2 (MH + ).
步驟3：4－(5,6－二甲氧基－1H－苯幷咪唑－2－基硫基甲基)－苯甲酸(189)： 依照與流程27步驟1(實例66及67)對烷基化反應所述相同之程序使化合物185與4－溴－甲苯甲酸反應獲得標題化合物189(產率60%)。1 H NMR：(DMSO－d6 )δ(ppm)：7.83(d,J＝8.2 Hz,2H),7.43(d,J＝8.4 Hz,2H),7.06(s,2H),4.61(s,2H)。m/z：345.2(MH＋ )。 Step 3: 4-(5,6-Dimethoxy-1H-benzimidazol-2-ylsulfanylmethyl)-benzoic acid (189): mp EtOAc (m.) The same procedure as described for the urelation reaction gave Compound 185 to 4-bromo-toluic acid to afford the title compound 189 (yield 60%). 1 H NMR: (DMSO-d 6 ) δ (ppm): 7.83 (d, J = 8.2 Hz, 2H), 7.43 (d, J = 8.4 Hz, 2H), 7.06 (s, 2H), 4.61 (s, 2H). m/z: 345.2 (MH + ).
步驟4：N－(2－胺基－苯基)－4－(5,6－二甲氧基－1H－苯幷咪唑－2－基硫基甲基)－苯甲醯胺(193) 依照與流程1步驟5(實例1)對BOP偶合反應所述相同之程序用化合物189作為原料獲得標題化合物193(148 mg，產率59%)。1 H NMR：(DMSO－d6 )δ(ppm)：12.30(s,1H),9.55(s,1H),7.85(d,J＝8.0 Hz,2H),7.48(d,J＝8.4 Hz,2H),7.10(d,J＝7.8 Hz,2H),6.92(td,J＝7.2,1.6 Hz,1H),6.91－6.85(bs,1H),6.73(dd,J＝8.2,1.2 Hz,1H),6.55(td,J＝7.8,1.6 Hz,1H),4.85(s,2H),4.52(s,2H),3.74(s,6H)。m/z：435.5(MH＋ )。 Step 4: N-(2-Amino-phenyl)-4-(5,6-dimethoxy-1H-benzoimidazol-2-ylthiomethyl)-benzamide (193) according to The title compound 193 (148 mg, yield 59%) was obtained from the title compound 189. 1 H NMR: (DMSO-d 6 ) δ (ppm): 12.30 (s, 1H), 9.55 (s, 1H), 7.85 (d, J = 8.0 Hz, 2H), 7.48 (d, J = 8.4 Hz, 2H), 7.10 (d, J = 7.8 Hz, 2H), 6.92 (td, J = 7.2, 1.6 Hz, 1H), 6.91-6.85 (bs, 1H), 6.73 (dd, J = 8.2, 1.2 Hz, 1H) ), 6.55 (td, J = 7.8, 1.6 Hz, 1H), 4.85 (s, 2H), 4.52 (s, 2H), 3.74 (s, 6H). m/z: 435.5 (MH + ).
N－(2－胺基－苯基)－4－(5,6－二氟－1H－苯幷咪唑－2－基硫基甲基)－苯甲醯胺(194)步驟1：4,5－二氟－苯－1,2－二胺(183)： 依照與實例92步驟1(流程38)所述相同之程序，但以4,5－二氟－2－硝基苯胺代替1,2－二甲氧基－4,5－二硝基苯獲得標題化合物183(產率97%)。1 H NMR：(CD3 OD)δ(ppm)：6.53(t,J＝10.0 Hz,2H)。m/z：145.3(MH＋ )。 N-(2-Amino-phenyl)-4-(5,6-difluoro-1H-benzoimidazol-2-ylthiomethyl)-benzamide (194) Step 1:4,5 -Difluoro-benzene-1,2-diamine (183): following the same procedure as described in Step 92 (Scheme 38) of Example 92, but replacing 1,2 with 4,5-difluoro-2-nitroaniline -Dimethoxy-4,5-dinitrobenzene gave the title compound 183 (yield: 97%). 1 H NMR: (CD 3 OD) δ (ppm): 6.53 (t, J = 10.0 Hz, 2H). m/z: 145.3 (MH + ).
步驟2：5,6－二氟－1H－苯幷咪唑－2－硫醇(186)： 依照J.Med.Chem.,1998,63,977－983所述之程序自化合物183開始獲得標題化合物186(產率60%)。1 H NMR：(CD3 OD)δ(ppm)：7.48(s,0.5H),7.13(d,J＝8.4 Hz,1H),7.11(d,J＝6.4 Hz,1H),1.99(s,1.5H)。m/z：187.1(MH＋ )。 Step 2: 5,6-Difluoro-1H-benzimidazole-2-thiol (186): The title compound 186 was obtained from compound 183 according to the procedure described in J. Med. Chem., 1998, 63, 977-983 ( Yield 60%). 1 H NMR: (CD 3 OD) δ (ppm): 7.48 (s, 0.5H), 7.13 (d, J = 8.4 Hz, 1H), 7.11 (d, J = 6.4 Hz, 1H), 1.99 (s, 1.5H). m/z: 187.1 (MH + ).
步驟3：4－(5,6－二氟－1H－苯幷咪唑－2－基硫基甲基)－苯甲酸(190)： 依照與流程27步驟1(實例66及67)對烷基化反應所述相同之程序使化合物186與4－溴－甲苯甲酸反應獲得標題化合物190(產率59%)。1 H NMR：(DMsO－d6 )δ(ppm)：9.07(s,0.5H),7.84(d,J＝8.0 Hz,2H),7.68(s,1.5H),7.52(d,J＝8.2 Hz,2H),5.53－5.45(m,2H),4.60(s,2H)。m/z：321.2(MH＋ )。 Step 3: 4-(5,6-Difluoro-1H-benzimidazol-2-ylthiomethyl)-benzoic acid (190): alkylation according to Step 1 (Examples 66 and 67) of Scheme 27 The same procedure was followed for the reaction of Compound 186 with 4-bromo-toluic acid to give the title compound 190 (yield: 59%). 1 H NMR: (DMsO-d 6 ) δ (ppm): 9.07 (s, 0.5H), 7.84 (d, J = 8.0 Hz, 2H), 7.68 (s, 1.5H), 7.52 (d, J = 8.2 Hz, 2H), 5.53-5.45 (m, 2H), 4.60 (s, 2H). m/z: 321.2 (MH + ).
步驟4：N－(2－胺基－苯基)－4－(5,6－二氟－1H－苯幷咪唑－2－基硫基甲基)－苯甲醯胺(194) 依照與流程1步驟5(實例1)對BOP偶合反應所述相同之程序用化合物186作為原料獲得標題化合物194(產率39%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.59(s,1H),7.88(d,J＝8.0 Hz,2H),7.54(d,J＝8.0 Hz,2H),7.55－7.40(m,2H),7.13(d,J＝7.6 Hz,1H),6.95(t,J＝7.6 Hz,1H),6.76(d,J＝7.4 Hz,1H),6.58(t,J＝7.4 Hz,1H),4.61(s,2H)。m/z：411.4(MH＋ )。 Step 4: N-(2-Amino-phenyl)-4-(5,6-difluoro-1H-benzoimidazol-2-ylthiomethyl)-benzamide (194) according to the procedure 1 Step 5 (Example 1) The same procedure as for the BOP coupling reaction The title compound 194 (yield 39%) was obtained. 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.59 (s, 1H), 7.88 (d, J = 8.0 Hz, 2H), 7.54 (d, J = 8.0 Hz, 2H), 7.55-7.40 ( m, 2H), 7.13 (d, J = 7.6 Hz, 1H), 6.95 (t, J = 7.6 Hz, 1H), 6.76 (d, J = 7.4 Hz, 1H), 6.58 (t, J = 7.4 Hz, 1H), 4.61 (s, 2H). m/z: 411.4 (MH + ).
N－(2－胺基－苯基)－4－(5－氯－6－氟－1H－苯幷咪唑－2－基硫基甲基)－苯甲醯胺(195)步驟1：4－(6－氯－5－氟－1H－苯幷咪唑－2－基硫基甲基)－苯甲酸甲酯(187) 依照與流程27步驟1(實例66及67)對烷基化反應所述相同之程序使6－氯－5－氟苯幷咪唑－2－硫醇與4－(溴甲基)苯甲酸甲酯反應獲得標題化合物187(產率54%)。1 H NMR：(DMSO－d6 )δ(ppm)：(母峰,丟失質子：7.85(d,J＝8.4 Hz,2H),7.55(d,J＝8.4 Hz,2H),3.80(s,2H),3.34(s,3H)。m/z：351.2(MH＋ )。 N-(2-Amino-phenyl)-4-(5-chloro-6-fluoro-1H-benzoimidazol-2-ylthiomethyl)-benzamide (195) Step 1: 4- (6-Chloro-5-fluoro-1H-benzimidazol-2-ylthiomethyl)-benzoic acid methyl ester (187) as described for the alkylation reaction with Step 27 (Examples 66 and 67) of Scheme 27 The same procedure was used to react 6-chloro-5-fluorobenzimidazole-2-thiol with methyl 4-(bromomethyl)benzoate to give the title compound 187 (yield 54%). 1 H NMR: (DMSO-d 6 ) δ (ppm): (mother peak, lost proton: 7.85 (d, J = 8.4 Hz, 2H), 7.55 (d, J = 8.4 Hz, 2H), 3.80 (s, 2H), 3.34 (s, 3H), m/z: 351.2 (MH + ).
步驟2：4－(5－氯－6－氟－1H－苯幷咪唑－2－基硫基甲基)－苯甲酸(191)： 依照與流程1步驟4(實例1)對水解所述相同之程序用化合物187作為原料獲得標題化合物191(產率83%)。1 H NMR：(DMSO－d6 )δ(ppm)：7.88(d,J＝8.2 Hz,2H),7.67(d,J＝6.8 Hz,1H),7.55(d,J＝8.2 Hz,2H),7.53(d,J＝6.8 Hz,1H),4.65(s,2H)。m/z：337.2(MH＋ )。 Step 2: 4-(5-Chloro-6-fluoro-1H-benzimidazol-2-ylthiomethyl)-benzoic acid (191): identical to that described for the hydrolysis in step 1 of Example 1 (Example 1) The title compound 191 (yield: 83%) was obtained from Compound 187. 1 H NMR: (DMSO-d 6 ) δ (ppm): 7.88 (d, J = 8.2 Hz, 2H), 7.67 (d, J = 6.8 Hz, 1H), 7.55 (d, J = 8.2 Hz, 2H) , 7.53 (d, J = 6.8 Hz, 1H), 4.65 (s, 2H). m/z: 337.2 (MH + ).
步驟3：N－(2－胺基－苯基)－4－(5－氯－6－氟－1H－苯幷咪唑－2－基硫基甲基)－苯甲醯胺(195) 依照與流程1步驟5(實例1)對BOP偶合反應所述相同之程序用化合物191作為原料獲得標題化合物195(產率62%)。1 H NMR：(DMSO－d6 )δ(ppm)：12.87(bs,1H),9.56(s,1H),7.87(d,J＝8.0 Hz,2H),7.62－7.57(m,1H),7.53(d,J＝8.2 Hz,2H),7.52－7.48(m,1H),7.10(d,J＝7.8 Hz,1H),6.92(td,J＝8.0,1.6 Hz,1H),6.73(dd,J＝7.8,1.4 Hz,1H),6.55(t,J＝7.4 Hz,1H),4.86(s,2H),4.61(s,2H)。m/z：427.4(MH＋ )。 Step 3: N-(2-Amino-phenyl)-4-(5-chloro-6-fluoro-1H-benzoimidazol-2-ylthiomethyl)-benzamide (195) according to Procedure 1 Step 5 (Example 1) For the BOP coupling reaction, the same procedure was used to obtain the title compound 195 (yield: 62%). 1 H NMR: (DMSO-d 6 ) δ (ppm): 12.87 (bs, 1H), 9.56 (s, 1H), 7.78 (d, J = 8.0 Hz, 2H), 7.62-7.57 (m, 1H), 7.53 (d, J = 8.2 Hz, 2H), 7.52-7.48 (m, 1H), 7.10 (d, J = 7.8 Hz, 1H), 6.92 (td, J = 8.0, 1.6 Hz, 1H), 6.73 (dd , J = 7.8, 1.4 Hz, 1H), 6.55 (t, J = 7.4 Hz, 1H), 4.86 (s, 2H), 4.61 (s, 2H). m/z: 427.4 (MH + ).
N－(2－羥基－苯基)－4－(5－甲氧基－1H－苯幷咪唑－2－基硫基甲基)－苯甲醯胺(199)步驟1：2－(第三丁基－二甲基－矽烷氧基)－苯胺(196)： 向2－胺基苯酚(3.00 g，27.5 mmol)之DCM(150 ml)經攪拌溶液中加入第三丁基二甲基氯化矽(4.35 ml，28.9 mmol)及Et3 N(4.02 ml，28.9 mmol)。在室溫下攪拌該反應混合物16小時。以水及鹽水洗滌該有機層，使其經無水MgSO4 乾燥，過濾及濃縮。由急驟層析法(在己烷中5% AcOEt)純化該殘餘物產生標題化合物196(5.56 g，產率91%)。1 HNMR(CDCl3 )δ(ppm)：7.61(s,1H),7.16(s,1H),6.58(s,2H),6.45(s,1H),6.09(s,1H),3.97(s,3H),3.93(s,3H),3.84(s,3H),3.83(s,6H)。m/z：224.1(MH＋ )。 N-(2-hydroxy-phenyl)-4-(5-methoxy-1H-benzoimidazol-2-ylthiomethyl)-benzamide (199) Step 1: 2- (third Butyl-dimethyl-decyloxy)-aniline (196): To a solution of 2-aminophenol (3.00 g, 27.5 mmol) in DCM (150 ml)矽 (4.35 ml, 28.9 mmol) and Et 3 N (4.02 ml, 28.9 mmol). The reaction mixture was stirred at room temperature for 16 hours. Washed with water and brine the organic layer, it was dried over anhydrous MgSO 4, filtered and concentrated. The residue was purified by flash chromatography eluting elut elut elut elut elut 1 H NMR (CDCl 3 ) δ (ppm): 7.61 (s, 1H), 7.16 (s, 1H), 6.58 (s, 2H), 6.45 (s, 1H), 6.09 (s, 1H), 3.97 (s, 3H), 3.93 (s, 3H), 3.84 (s, 3H), 3.83 (s, 6H). m/z: 224.1 (MH + ).
步驟2：N－[2－(第三丁基－二甲基－矽烷氧基)－苯基]－4－(5－甲氧基－1H－苯幷咪唑－2－基硫基甲基)－苯甲醯胺(198)： 依照與流程1步驟5(實例1)對BOP偶合反應所述相同之程序使化合物197(描述於專利申請案WO 03/024448)與化合物196反應獲得標題化合物198。m/z：520.3(MH＋ )。 Step 2: N-[2-(Tertiary butyl-dimethyl-decyloxy)-phenyl]-4-(5-methoxy-1H-benzoimidazol-2-ylthiomethyl) - Benzamide (198): Compound 197 (described in patent application WO 03/024448) was reacted with compound 196 to give the title compound 198 according to the same procedure as described for the </ br> . m/z: 520.3 (MH + ).
步驟3：N－(2－羥基－苯基)－4－(5－甲氧基－1H－苯幷咪唑－2－基硫基甲基)－苯甲醯胺(199) 向化合物198(313 mg，0.600 mmol)之THF(15 ml)經攪拌溶液中加入溶於THF(1.20 ml，1.20 mmol)中之TBAF 1 M。在室溫下攪拌該反應混合物16小時。蒸發溶劑且將殘餘物溶解於EtOAc中，以飽和NH4 Cl及鹽水洗滌，將其經無水MgSO4 乾燥，接著過濾且加以濃縮產生呈白色粉末之標題化合物199 (150 mg，產率61%)。1 H NMR(DMSO－d6 )δ(ppm)：9.72(bs,1H),9.49(bs,1H),7.90(d,J＝8.1 Hz,2H),7.63(d,J＝8.1 Hz,1H),7.58(d,J＝8.8 Hz,2H),7.51(d,J＝9.5 Hz,1H),7.08－6.89(m,4H),6.81(dd,J＝7.0,7.0 Hz,1H),4.76(s,2H),3.81(s,3H)。m/z：406.2(MH＋ )。 Step 3: N-(2-Hydroxy-phenyl)-4-(5-methoxy-1H-benzoimidazol-2-ylthiomethyl)-benzamide (199) to compound 198 (313 To a stirred solution was added TBAF 1 M in THF (1.20 ml, 1.20 mmol). The reaction mixture was stirred at room temperature for 16 hours. The solvent was evaporated and the residue was dissolved in EtOAc, washed with saturated NH 4 Cl and brine, dried it over anhydrous MgSO 4, then filtered and be concentrated to give the title as a white powder of compound 199 (150 mg, 61% yield) . 1 H NMR (DMSO-d 6 ) δ (ppm): 9.72 (bs, 1H), 9.49 (bs, 1H), 7.90 (d, J = 8.1 Hz, 2H), 7.63 (d, J = 8.1 Hz, 1H) ), 7.58 (d, J = 8.8 Hz, 2H), 7.51 (d, J = 9.5 Hz, 1H), 7.08-6.89 (m, 4H), 6.81 (dd, J = 7.0, 7.0 Hz, 1H), 4.76 (s, 2H), 3.81 (s, 3H). m/z: 406.2 (MH + ).
N－(2－羥基－苯基)－4－{[6－(2－嗎啉－4－基－乙氧基)－苯幷噻唑－2－基胺基]－甲基}－苯甲醯胺(201) 依照與實例95所述相同之程序以化合物200 (描述於專利WO 03/024448中)代替化合物197且在最後一步中以1 N HCl代替TBAF獲得標題化合物201(產率26%)。1 H NMR：(CD3 OD)δ(ppm)：7.93(d,J＝8.5 Hz,2H),7.79(d,J＝7.5 Hz,1H),7.55(d,J＝8.0 Hz,2H),7.33(d,J＝8.5 Hz,1H),7.23(s,1H),7.04(t,J＝7.0 Hz,1H),6.92－6.85(m,3H),7.40(s,2H),4.14－4.12(m,2H),3.72－3.70(m,4H),2.81－2.79(m,2H),2.62－2.60(m,4H)。m/z：505.5(MH＋ )。 N-(2-hydroxy-phenyl)-4-{[6-(2-morpholin-4-yl-ethoxy)-benzothiazol-2-ylamino]-methyl}-benzamide The amine (201) was replaced by the same procedure as described in Example 95 in the compound 200 (described in the patent WO 03/024448) in place of the compound 197 and in the last step, 1 N HCl was used in place of TBAF to obtain the title compound 201 (yield 26%). . 1 H NMR: (CD 3 OD) δ (ppm): 7.93 (d, J = 8.5 Hz, 2H), 7.79 (d, J = 7.5 Hz, 1H), 7.55 (d, J = 8.0 Hz, 2H), 7.33 (d, J = 8.5 Hz, 1H), 7.23 (s, 1H), 7.04 (t, J = 7.0 Hz, 1H), 6.92 - 6.85 (m, 3H), 7.40 (s, 2H), 4.14 - 4.12 (m, 2H), 3.72-3.70 (m, 4H), 2.81-2.79 (m, 2H), 2.62-2.60 (m, 4H). m/z: 505.5 (MH + ).
N－(2－胺基－苯基)－4－[3－(吡啶－3－基胺基)－吡咯啶－1－基]－苯甲醯胺(206)步驟1：4－(3－羥基－吡咯啶－1－基)－苯甲酸第三丁酯(202)： 依照J.Heterocycl.Chem.,1994,31,1241所述之程序獲得標題化合物202(產率91%)。1 H NMR：(CD3 OD)δ(ppm)：7.77(d,J＝9.0 Hz,2H),6.54(d,J＝9.0 Hz,2H),4.57－4.53(m,1H),3.57－3.50(m,2H),3.45(td,J＝9.4,3.3 Hz,1H),3.29(dd,J＝12.7,1.6 Hz,1H),2.22－2.13(m,1H),2.10－2.03(m,1H),1.59(s,9H)。m/z：264.4(MH＋ )。 N-(2-Amino-phenyl)-4-[3-(pyridin-3-ylamino)-pyrrolidin-1-yl]-benzamide (206) Step 1: 4-(3- Hydroxy-pyrrolidin-1-yl)-benzoic acid tert-butyl ester (202): The title compound 202 (yield: 91%) was obtained according to the procedure of J. Heterocycl. Chem., 1994, 31, 1241. 1 H NMR: (CD 3 OD) δ (ppm): 7.77 (d, J = 9.0 Hz, 2H), 6.54 (d, J = 9.0 Hz, 2H), 4.57 - 4.53 (m, 1H), 3.57 - 3.50 (m, 2H), 3.45 (td, J = 9.4, 3.3 Hz, 1H), 3.29 (dd, J = 12.7, 1.6 Hz, 1H), 2.22 - 2.13 (m, 1H), 2.10-2.03 (m, 1H) ), 1.59 (s, 9H). m/z: 264.4 (MH + ).
步驟2：4－(3－酮基－吡咯啶－1－基)－苯甲酸第三丁酯(203)： 依照J.Heterocycl.Chem.,1994,31,1241所述之程序獲得標題化合物203(產率73)。1 H NMR：(DMSO－d6 )δ(ppm)：7.74(d,J＝8.8 Hz,2H),6.67(d,J＝9.0 Hz,2H),3.75(s,2H),3.69(t,J＝7.4 Hz,2H),2.72(t,J＝7.6 Hz,2H),1.52(s,9H)。m/z：262.4(MH＋ )。 Step 2: 4-(3-keto-pyrrolidin-1-yl)-benzoic acid tert-butyl ester (203): The title compound 203 was obtained according to the procedure of J. Heterocycl. Chem., 1994, 31, 1241. (Yield 73). 1 H NMR: (DMSO-d 6 ) δ (ppm): 7.74 (d, J = 8.8 Hz, 2H), 6.67 (d, J = 9.0 Hz, 2H), 3.75 (s, 2H), 3.69 (t, J = 7.4 Hz, 2H), 2.72 (t, J = 7.6 Hz, 2H), 1.52 (s, 9H). m/z: 262.4 (MH + ).
步驟3：4－[3－(吡啶－3－基胺基)－吡咯啶－1－基]－苯甲酸第三丁酯(204)： 依照與流程3步驟2(實例12)對還原胺化反應所述相同之程序自化合物203開始且使用3－胺基吡啶代替6－(吡啶－3－基)吡啶－2－胺(11)獲得標題化合物204(產率76%)。1 H NMR：(丙酮－d6 )δ(ppm)：8.08(d,J＝2.7 Hz,1H),7.85(dd,J＝4.3,1.6 Hz,1H),7.79(d,J＝9.0 Hz,2H),7.09(ddd,J＝8.2,4.5,0.8 Hz,1H),7.06(ddd,J＝8.2,2.7,1.6 Hz,1H),6.58(d,J＝9.0 Hz,2H),4.33(五重峰,J＝4.9 Hz,1H),3.79(dd,J＝10.2,6.1 Hz,1H),3.60－3.54(m,1H),3.51－3.45(m,1H),3.32(dd,J＝7.2,4.1 Hz,1H),2.44(六重峰,J＝7.6 Hz,1H),2.18－2.11(m,1H),1.56(s,9H)。m/z：340.4(MH＋ )。 Step 3: 4-[3-(Pyridin-3-ylamino)-pyrrolidin-1-yl]-benzoic acid tert-butyl ester (204): reductive amination according to procedure 2, step 2 (Example 12) The same procedure as the reaction was carried out starting from compound 203 and using 3-aminopyridine instead of 6-(pyridin-3-yl)pyridin-2-amine (11) afforded the title compound 204 (yield 76%). 1 H NMR: (acetone-d 6 ) δ (ppm): 8.08 (d, J = 2.7 Hz, 1H), 7.85 (dd, J = 4.3, 1.6 Hz, 1H), 7.79 (d, J = 9.0 Hz, 2H), 7.09 (ddd, J=8.2, 4.5, 0.8 Hz, 1H), 7.06 (ddd, J=8.2, 2.7, 1.6 Hz, 1H), 6.58 (d, J=9.0 Hz, 2H), 4.33 (five Heavy peak, J=4.9 Hz, 1H), 3.79 (dd, J=10.2, 6.1 Hz, 1H), 3.60-3.54 (m, 1H), 3.51-3.45 (m, 1H), 3.32 (dd, J=7.2 , 4.1 Hz, 1H), 2.44 (sixfold, J = 7.6 Hz, 1H), 2.18-2.11 (m, 1H), 1.56 (s, 9H). m/z: 340.4 (MH + ).
步驟4：4－[3－(吡啶－3－基胺基)－吡咯啶－1－基]－苯甲酸(205)： 依照與流程28步驟5(實例68)對Boc***所述相同之程序用化合物204作為原料獲得標題化合物205(產率96%)。1 H NMR：(DMSO－d6 )δ(ppm)：8.09(d,J＝2.0 Hz,1H),8.02(d,J＝3.5 Hz,1H),7.74(d,J＝8.8 Hz,2H),7.68－7.62(m,2H),7.29－7.26(m,1H),6.57(d,J＝8.8 Hz,2H),4.29－4.26(m,1H),3.71(dd,J＝10.6,5.7 Hz,1H),3.51－3.42(m,2H),3.23(dd,J＝10.6,3.5 Hz,1H),2.35(六重峰,J＝7.2 Hz,1H),2.06－1.98(m,1H)。m/z：284.4(MH＋ )。 Step 4: 4-[3-(Pyridin-3-ylamino)-pyrrolidin-1-yl]-benzoic acid (205): according to the same procedure as described for the Boc split in step 28 of Example 28 (Example 68) The title compound 205 (yield 96%) was obtained from Compound 204. 1 H NMR: (DMSO-d 6 ) δ (ppm): 8.09 (d, J = 2.0 Hz, 1H), 8.02 (d, J = 3.5 Hz, 1H), 7.74 (d, J = 8.8 Hz, 2H) , 7.68-7.62 (m, 2H), 7.29-7.26 (m, 1H), 6.57 (d, J = 8.8 Hz, 2H), 4.29-4.26 (m, 1H), 3.71 (dd, J = 10.6, 5.7 Hz , 1H), 3.51-3.42 (m, 2H), 3.23 (dd, J = 10.6, 3.5 Hz, 1H), 2.35 (six-peak, J = 7.2 Hz, 1H), 2.06-1.98 (m, 1H). m/z: 284.4 (MH + ).
步驟5：N－(2－胺基－苯基)－4－[3－(吡啶－3－基胺基)－吡咯啶－1－基]－苯甲醯胺(206) 依照與流程1步驟5(實例1)對BOP偶合反應所述相同之程序用化合物205作為原料獲得標題化合物206(產率10%)。1 H NMR：(CD3 OD)δ(ppm)：8.14(s,1H),7.96(d,J＝2.3 Hz,1H),7.86(d,J＝9.0 Hz,2H),7.77(dd,J＝4.7,1.4 Hz,1H),7.61－7.58(m,1H),7.25(dd,J＝6.1,3.1 Hz,1H),7.18(ddd,J＝8.4,4.7,0.8 Hz,1H),7.15(dd,J＝8.0,1.6 Hz,1H),7.11(ddd,J＝8.2,2.7,1.4 Hz,1H),7.05(td,J＝7.2,1.4 Hz,1H),6.89(dd,J＝8.0,1.4 Hz,1H),6.76(td,J＝7.6,1.4 Hz,1H),6.64(d,J＝8.8 Hz,2H),4.26(五重峰,J＝4.3 Hz,1H),3.78(dd,J＝10.0,6.1 Hz,1H),3.62－3.56(m,1H),3.53－3.47(m,1H),3.31－3.29(m,1H),2.42(六重峰,J＝7.4 Hz,1H),2.12－2.08(m,1H)。m/z：374.4(MH＋ )。 Step 5: N-(2-Amino-phenyl)-4-[3-(pyridin-3-ylamino)-pyrrolidin-1-yl]-benzamide (206) according to step 1 5 (Example 1) The same procedure as described for the BOP coupling reaction The title compound 206 (yield 10%) was obtained using Compound 205 as a starting material. 1 H NMR: (CD 3 OD) δ (ppm): 8.14 (s, 1H), 7.96 (d, J = 2.3 Hz, 1H), 7.86 (d, J = 9.0 Hz, 2H), 7.77 (dd, J = 4.7, 1.4 Hz, 1H), 7.61 - 7.58 (m, 1H), 7.25 (dd, J = 6.1, 3.1 Hz, 1H), 7.18 (ddd, J = 8.4, 4.7, 0.8 Hz, 1H), 7.15 ( Dd, J = 8.0, 1.6 Hz, 1H), 7.11 (ddd, J = 8.2, 2.7, 1.4 Hz, 1H), 7.05 (td, J = 7.2, 1.4 Hz, 1H), 6.89 (dd, J = 8.0, 1.4 Hz, 1H), 6.76 (td, J = 7.6, 1.4 Hz, 1H), 6.64 (d, J = 8.8 Hz, 2H), 4.26 (five peaks, J = 4.3 Hz, 1H), 3.78 (dd, J = 10.0, 6.1 Hz, 1H), 3.62-3.56 (m, 1H), 3.53-3.47 (m, 1H), 3.31-3.29 (m, 1H), 2.42 (sixfold, J = 7.4 Hz, 1H) , 2.12 - 2.08 (m, 1H). m/z: 374.4 (MH + ).
4－(2－胺基－苯幷噻唑－6－基氧基甲基)－N－(2－胺基－苯基)－苯甲醯胺(209)步驟1：{2－[4－(2－胺基－苯幷噻唑－6－基氧基)－苯甲醯胺基]－苯基}－胺基甲酸第三丁酯(208)： 依照與合成化合物133(流程32)所用相同之程序，用化合物207(描述於專利申請案WO 03/024448中)代替二甲胺基－乙醇且以化合物127(亦提及於流程32)代替化合物129(流程32)獲得標題化合物208(產率43%)。1 H NMR(DMSO－d6 )δ(ppm)：9.81(s,1H),8.66(s,1H),7.94(d,J＝8.4 Hz,2H),7.58(d,J＝8.4 Hz,2H),7.51(d,J＝8.2 Hz,2H),7.37(d,J＝2.5 Hz 1H),7.23(d,J＝8.8 Hz,1H),7.18(td,J＝7.8,1.8 Hz,1H),7.13(td,J＝7.6,1.6 Hz,1H),6.89(dd,J＝8.6,2.5 Hz,1H),5.18(s,2H),1.43(s,9H)。m/z：491.4(MH＋ )。 4-(2-Amino-benzothiazole-6-yloxymethyl)-N-(2-amino-phenyl)-benzamide (209) Step 1: {2-[4-( 2-Amino-benzothiazole-6-yloxy)-benzylaminoamido]-phenyl}-carbamic acid tert-butyl ester (208): identical to that used in the synthesis of compound 133 (Scheme 32) Procedure, using the compound 207 (described in patent application WO 03/024448) instead of dimethylamino-ethanol and the compound 127 (also referred to in Scheme 32) in place of compound 129 (Scheme 32) to give the title compound 208 (yield 43%). 1 H NMR (DMSO-d 6 ) δ (ppm): 9.81 (s, 1H), 8.66 (s, 1H), 7.94 (d, J = 8.4 Hz, 2H), 7.58 (d, J = 8.4 Hz, 2H ), 7.51 (d, J = 8.2 Hz, 2H), 7.37 (d, J = 2.5 Hz 1H), 7.23 (d, J = 8.8 Hz, 1H), 7.18 (td, J = 7.8, 1.8 Hz, 1H) , 7.13 (td, J = 7.6, 1.6 Hz, 1H), 6.89 (dd, J = 8.6, 2.5 Hz, 1H), 5.18 (s, 2H), 1.43 (s, 9H). m/z: 491.4 (MH + ).
步驟2：N－(2－胺基－苯基)－4－[3－(6－甲氧基－苯幷噻唑－2－基)－脲基]－苯甲醯胺(209) 依照與流程28步驟5(實例68)對Boc***所述相同之程序用化合物208作為原料獲得標題化合物209(產率28%)。1 H NMR：(DMSO－d6 )8(ppm)：9.63(s,1H),7.96(d,J＝8.2 Hz,2H),7.54(d,J＝8.2 Hz,2H),7.37(d,J＝2.5 Hz,1H),7.23(s,2H),7.21(d,J＝8.8 Hz,1H),7.14(d,J＝8.0 Hz,1H),6.95(td,J＝8.8,2.3 Hz,1H),6.89(dd,J＝8.6,2.5 Hz,1H),6.75(d,J＝6.7 Hz,1H),6.57(t,J＝6.7 Hz,1H),5.16(s,2H),4.89(s,2H)。m/z：391.4(MH＋ )。 Step 2: N-(2-Amino-phenyl)-4-[3-(6-methoxy-benzothiazol-2-yl)-ureido]-benzamide (209) according to the procedure 28 Step 5 (Example 68) The title compound 209 (yield: 28%) 1 H NMR: (DMSO-d 6 ) 8 (ppm): 9.63 (s, 1H), 7.96 (d, J = 8.2 Hz, 2H), 7.54 (d, J = 8.2 Hz, 2H), 7.37 (d, J = 2.5 Hz, 1H), 7.23 (s, 2H), 7.21 (d, J = 8.8 Hz, 1H), 7.14 (d, J = 8.0 Hz, 1H), 6.95 (td, J = 8.8, 2.3 Hz, 1H), 6.89 (dd, J = 8.6, 2.5 Hz, 1H), 6.75 (d, J = 6.7 Hz, 1H), 6.57 (t, J = 6.7 Hz, 1H), 5.16 (s, 2H), 4.89 ( s, 2H). m/z: 391.4 (MH + ).
N－(2－胺基－苯基)－4－[(4－甲磺醯基－苯胺基)－甲基]－苯甲醯胺(212a)步驟1：4－[(4－甲磺醯基－苯胺基)－甲基]－苯甲酸(211a) 依照流程3步驟2(實例12)所述之程序，藉由使4－甲磺醯基－苯胺基(210a)與4－甲醯基－苯甲酸反應獲得該標題化合物。1 H NMR,(DMSO)δ(ppm)：7.87(d,J＝7.6 Hz,2H),7.50(d,J＝8.2 Hz,2H),7.41(d,J＝7.6 Hz,2H),6.64(d,J＝8.0 Hz,2H),4.42(s,2H),3.00(s,3H)。LRMS：(計算值)305.4；(獲得值)304.3(MH)＋ 。 N-(2-Amino-phenyl)-4-[(4-methylsulfonyl-anilino)-methyl]-benzamide (212a) Step 1: 4-[(4-Methanesulfonate) Benzyl-anilino)-methyl]-benzoic acid (211a) according to the procedure described in Scheme 3, Step 2 (Example 12), by 4-methylsulfonyl-anilino (210a) and 4-methylindenyl - Benzoic acid reaction to give the title compound. 1 H NMR, (DMSO) δ (ppm): 7.87 (d, J = 7.6 Hz, 2H), 7.50 (d, J = 8.2 Hz, 2H), 7.41 (d, J = 7.6 Hz, 2H), 6.64 ( d, J = 8.0 Hz, 2H), 4.42 (s, 2H), 3.00 (s, 3H). LRMS: (calculated) 305.4; (obtained value) 304.3 (MH) + .
步驟2：N－(2－胺基－苯基)－4－[(4－甲磺醯基－苯胺基)－甲基]－苯甲醯胺(212a) 依照流程1步驟5(實例1)所述之程序，藉由使酸211a與1,2－苯二胺反應獲得該化合物。1 H NMR：(DMSO)δ(ppm)：9.57(bs,1H),7.90(d,J＝8.4 Hz,1H),7.50(d,J＝8.2 Hz,2H),7.43(d,J＝8.4 Hz,2H),7.35(t,J＝6.0 Hz,1H),7.11(d,J＝7.6 Hz,1H),6.93(dt,J＝1.6,8.0 Hz,1H),6.73(dd,J＝1.6,8.0 Hz,1H),6.66(d,J＝8.8 Hz,2H),6.55(dt,J＝1.2,7.6 Hz,1H),4.88(bs,2H),4.43(d,J＝6.0 Hz,2H)。LRMS：(計算值)395.5；(獲得值)396.4(MH)＋ 。 Step 2: N-(2-Amino-phenyl)-4-[(4-methylsulfonyl-anilino)-methyl]-benzamide (212a) according to Scheme 1 Step 5 (Example 1) The procedure described is carried out by reacting acid 211a with 1,2-phenylenediamine. 1 H NMR: (DMSO) δ (ppm): 9.57 (bs, 1H), 7.90 (d, J = 8.4 Hz, 1H), 7.50 (d, J = 8.2 Hz, 2H), 7.43 (d, J = 8.4 Hz, 2H), 7.35 (t, J = 6.0 Hz, 1H), 7.11 (d, J = 7.6 Hz, 1H), 6.93 (dt, J = 1.6, 8.0 Hz, 1H), 6.73 (dd, J = 1.6) , 8.0 Hz, 1H), 6.66 (d, J = 8.8 Hz, 2H), 6.55 (dt, J = 1.2, 7.6 Hz, 1H), 4.88 (bs, 2H), 4.43 (d, J = 6.0 Hz, 2H ). LRMS: (calculated) 395.5; (obtained) 396.4 (MH) + .
用對化合物212a實例99(流程42，表1)所述相同之程序自芳基胺210b－o經由中間體酸211b－o(流程42)製備實例100－113(化合物212b－o)。Example 100-113 (Compound 212b-o) was prepared from the arylamine 210b-o via intermediate acid 211b-o (Scheme 42) using the same procedure as described for compound 212a, Example 99 (Scheme 42, Table 1).
N－(2－胺基－苯基)－4－羥甲基－苯甲醯胺(213) 在一含有4－甲醯基苯甲酸(300 mg，1.8 mmol)之燒瓶中加入氯化二丁基錫(55 mg，0.18 mmol)，隨後加入THF(5 ml)及苯矽烷(0.187 ml，1.8 mmol)。將所得混合物在室溫氮氣下攪拌隔夜，濃縮且未經進一步純化即用於下一步(與鄰苯二胺偶合)，依照流程1步驟5(實例1)所述之程序產生化合物213(378 mg，產率78%)。1 H NMR：(DMSO)δ(ppm)：9.63(s,1H),7.94(d,J＝8.0 Hz,2H),7.43(d,J＝8.5 Hz,2H),7.16(d,J＝7.5 Hz,1H),6.96(t,J＝7.0 Hz,1H),6.78(d,J＝6.5,1H),6.59(dd,J＝7.0,7.5 Hz,1H),4.88(s,1H),4.57(s,2H)。LRMS：(計算值)242.3；(獲得值)243.4(MH)＋ 。 N-(2-Amino-phenyl)-4-hydroxymethyl-benzamide (213) was added to a flask containing 4-methylmercaptobenzoic acid (300 mg, 1.8 mmol) with dibutyltin chloride. (55 mg, 0.18 mmol), followed by THF (5 ml) and benzene hexane (0.187 ml, 1.8 mmol). The resulting mixture was stirred at room temperature under nitrogen overnight, concentrated and applied to the next step (coupling with o-phenylenediamine) without further purification to give compound 213 (378 mg) according to procedure of procedure 1 step 5 (example 1). , yield 78%). 1 H NMR: (DMSO) δ (ppm): 9.63 (s, 1H), 7.94 (d, J = 8.0 Hz, 2H), 7.43 (d, J = 8.5 Hz, 2H), 7.16 (d, J = 7.5) Hz, 1H), 6.96 (t, J = 7.0 Hz, 1H), 6.78 (d, J = 6.5, 1H), 6.59 (dd, J = 7.0, 7.5 Hz, 1H), 4.88 (s, 1H), 4.57 (s, 2H). LRMS: (calculated) 242.3; (obtained value) 243.4 (MH) + .
N－(2－胺基－苯基)－4－[(3－氟－4－嗎啉－4－基－苯胺基)－甲基]－苯甲醯胺(218a)步驟1：4－(2－氟－4－硝基－苯基)－嗎啉(215a) 在室溫下向214a(3 g，18.85 mmol)之DMF(20 mL)溶液中加入嗎啉(1.6 ml，18.85 mmol)及K2 CO3 (10.4 g，75.4 mmol)。在60℃下加熱該反應混合物16小時，將其冷卻，過濾且在真空下濃縮之。以AcOEt/己烷(40：60)為溶離劑經矽膠由急驟層析法純化殘餘物產生呈白色固體之標題化合物215a(4.0 g，產率89%)。LRMS：226.2(計算值)；227.3(獲得值)(MH)＋ 。 N-(2-Amino-phenyl)-4-[(3-fluoro-4-morpholin-4-yl-anilino)-methyl]-benzamide (218a) Step 1: 4-( 2-Fluoro-4-nitro-phenyl)-morpholine (215a) was added morpholine (1.6 ml, 18.85 mmol) to 214a (3 g, 18.85 mmol) in DMF (20 mL) K 2 CO 3 (10.4 g, 75.4 mmol). The reaction mixture was heated at 60 <0>C for 16 h, cooled, filtered and concentrated in vacuo. The title compound 215a (4.0 g, yield: 89%) eluted elute elute LRMS: 226.2 (calculated); 227.3 (obtained) (MH) + .
步驟2：3－氟－4－嗎啉－4－基－苯胺(216a) 依照流程38步驟1(實例92)所述之程序，藉由催化氫化硝基化合物215a獲得標題化合物216a(產率92%)。LRMS：196.2(計算值)；197.2(獲得值)(MH)＋ 。 Step 2: 3-Fluoro-4-morpholin-4-yl-phenylamine (216a) The title compound 216a was obtained by catalytic hydrogenation of nitro compound 215a (yield 92) %). LRMS: 196.2 (calculated); 197.2 (obtained) (MH) + .
步驟3：4－[(3－氟－4－嗎啉－4－基－苯胺基)－甲基]－苯甲酸(217a) 依照流程3步驟2(實例12)所述之程序，經由使4－甲醯基苯甲酸酯與胺216a反應獲得標題化合物226a(產率91%)。LRMS：330.4(計算值)；331.5(獲得值)(MH)＋ 。 Step 3: 4-[(3-Fluoro-4-morpholin-4-yl-anilino)-methyl]-benzoic acid (217a) according to the procedure described in Step 3 (Example 12) of Scheme 3, via 4 -Methylmercaptobenzoate was reacted with amine 216a to give the title compound 226a (yield: 91%). LRMS: 330.4 (calculated); 331.5 (obtained) (MH) + .
步驟4：N－(2－胺基－苯基)4－[(3－氟－4－嗎啉－4－基－苯胺基)－甲基]－苯甲醯胺(218a) 依照流程1步驟5(實例1)使酸217a與1,2－苯二胺反應獲得標題化合物218a(產率40%)。1 H NMR：(DMSO)δ(ppm)：9.57(s,1H),7.91(d,J＝7.9 Hz,2H),7.44(d,J＝7.9 Hz,2H),7.14(d,J＝7.5 Hz,1H),6.95(t,J＝7.5；7.0 Hz,1H),6.82－6.74(m,2H),6.57(dd,J＝7.0；7.5 Hz,1H),6.37－6.30(m,2H),4.86(bs,2H),4.30(d,J＝5.71 Hz,2H),3.66(bs,4H),2.80(bs,4H)。LRMS：(計算值)420.2；(獲得值)421.2(MH)＋ 。 Step 4: N-(2-Amino-phenyl)4-[(3-fluoro-4-morpholin-4-yl-anilino)-methyl]-benzamide (218a) according to procedure 1 5 (Example 1) The acid 217a was reacted with 1,2-phenylenediamine to give the title compound 218a (yield 40%). 1 H NMR: (DMSO) δ (ppm): 9.57 (s, 1H), 7.91 (d, J = 7.9 Hz, 2H), 7.44 (d, J = 7.9 Hz, 2H), 7.14 (d, J = 7.5) Hz, 1H), 6.95 (t, J = 7.5; 7.0 Hz, 1H), 6.82-6.74 (m, 2H), 6.57 (dd, J = 7.0; 7.5 Hz, 1H), 6.37-6.30 (m, 2H) , 4.86 (bs, 2H), 4.30 (d, J = 5.71 Hz, 2H), 3.66 (bs, 4H), 2.80 (bs, 4H). LRMS: (calculated) 420.2; (obtained) 421.2 (MH) + .
依照與對化合物218a(實例116，流程44)(表3)所述相同之程序製備實例116－117(化合物218b－c)。Examples 116-117 (compounds 218b-c) were prepared according to the same procedure as described for compound 218a (Example 116, Scheme 44) (Table 3).
N－(2－胺基－苯基)－4－{[4－甲氧基－3－(2－嗎啉－4－基－乙氧基)－苯胺基]－甲基)－苯甲醯胺(222)步驟1：4－甲氧基－3－(2－嗎啉－4－基－乙氧基)－苯胺(220) 在室溫下向溶於吡啶(5 mL)及DMF(15 ml)之溶劑混合物中的4－(2－氯－乙基)－嗎啉(2.67 g，14.4 mmol)溶液中加入胺219(2.00，14.4 mmol)及K2 CO3 (7.96 g，57.6 mmol)。將該反應混合物在60℃下加熱隔夜，冷卻，過濾且在真空下濃縮之。由急驟層析法經矽膠以70：30 AcOEt/己烷溶離來純化該殘餘物產生標題化合物229(3.6 g，產率100%)。LRMS：252.3(計算值)；253.3(獲得值)(MH)＋ 。 N-(2-Amino-phenyl)-4-{[4-methoxy-3-(2-morpholin-4-yl-ethoxy)-anilino]-methyl)-benzamide Amine (222) Step 1: 4-methoxy-3-(2-morpholin-4-yl-ethoxy)-phenylamine (220) is dissolved in pyridine (5 mL) and DMF (15 ) at room temperature A solution of 4-(2-chloro-ethyl)-morpholine (2.67 g, 14.4 mmol) in a solvent mixture of ml) was added MeOH (2.00, 14.4 mmol) and K 2 CO 3 (7.96 g, 57.6 mmol) . The reaction mixture was heated at 60 <0>C overnight, cooled, filtered and concentrated in vacuo. The residue was purified by flash chromatography eluting elut elut elut elut elut elut elut elut LRMS: 252.3 (calculated); 253.3 (obtained) (MH) + .
步驟2：4－{[4－甲氧基－3－(2－嗎啉－4－基－乙氧基)－苯胺基]－甲基}－苯甲酸(221) 依照流程3步驟2(實例12)所述之程序使4－甲醯基苯甲酸酯與胺220反應獲得標題化合物221(1.9 g，產率99%)。LRMS：386.4(計算值)；387.4(獲得值)(MH)＋ 。 Step 2: 4-{[4-Methoxy-3-(2-morpholin-4-yl-ethoxy)-anilino]-methyl}-benzoic acid (221) according to Scheme 3, Step 2 (Example) 12) The procedure described was carried out by reacting 4-methylmercaptobenzoate with amine 220 to give the title compound 221 (1.9 g, yield 99%). LRMS: 386.4 (calculated); 387.4 (obtained) (MH) + .
步驟3：N－(2－胺基－苯基)－4－{[4－甲氧基－3－(2－嗎啉－4－基－乙氧基)－苯胺基]－甲基}－苯甲醯胺(222) 根據流程1步驟5(實例1)所述之程序藉由使酸221(5.07 mmol)與1,2－苯二胺(5.07 mmol)偶合獲得標題化合物222(260 mg，產率11%)。1 H NMR：(DMSO)δ(ppm)：9.59(s,1H),7.92(d,J＝7.5 Hz,2H),7.45(d,J＝7.9 Hz,2H),7.15(d,J＝7.9 Hz,1H),6.96(dd,J＝8.5；6.5 Hz,1H),6.77(d,J＝8.5 Hz,1H),6.65(d,J＝8.5 Hz,1H),6.58(t,J＝7.5 Hz,1H),6.31(d,J＝2.5 Hz,1H),6.03(d,J＝8.5 Hz,1H),4.86(bs,2H),4.30(d,J＝5.5 Hz,2H),3.95(dd,J＝5.9,5.5 Hz,2H),3.59(s,3H),3.56(bs,4H),2.63(bs,2H),2.44(bs,4H)。LRMS：476.6(計算值)；477.6(獲得值)(MH)＋ 。 Step 3: N-(2-Amino-phenyl)-4-{[4-methoxy-3-(2-morpholin-4-yl-ethoxy)-anilino]-methyl}- Benzalamine (222) was obtained by coupling the acid 221 (5.07 mmol) with 1,2-phenylenediamine (5.07 mmol ) according to the procedure of the procedure of Step 1 (Example 1) to give the title compound 222 (260 mg, Yield 11%). 1 H NMR: (DMSO) δ (ppm): 9.59 (s, 1H), 7.92 (d, J = 7.5 Hz, 2H), 7.45 (d, J = 7.9 Hz, 2H), 7.15 (d, J = 7.9) Hz, 1H), 6.96 (dd, J = 8.5; 6.5 Hz, 1H), 6.77 (d, J = 8.5 Hz, 1H), 6.65 (d, J = 8.5 Hz, 1H), 6.58 (t, J = 7.5) Hz, 1H), 6.31 (d, J = 2.5 Hz, 1H), 6.03 (d, J = 8.5 Hz, 1H), 4.86 (bs, 2H), 4.30 (d, J = 5.5 Hz, 2H), 3.95 ( Dd, J = 5.9, 5.5 Hz, 2H), 3.59 (s, 3H), 3.56 (bs, 4H), 2.63 (bs, 2H), 2.44 (bs, 4H). LRMS: 476.6 (calculated); 477.6 (obtained) (MH) + .
N－(2－胺基－苯基)－4－(3,4－二甲氧基－苯基胺磺醯基)－苯甲醯胺(224)步驟1：N－(3,4－二甲氧基－苯基)－4－碘－苯磺醯胺(223) 依照專利申請案第WO 01/38322 A1號所述之程序藉由使3,4－二甲氧基－苯胺與4－碘－苯磺醯氯反應獲得標題化合物223(產率80%)。LRMS：419.2(計算值)；420.2(獲得值)(MH)＋ 。 N-(2-Amino-phenyl)-4-(3,4-dimethoxy-phenylamine sulfonyl)-benzamide (224) Step 1: N-(3,4-di Methoxy-phenyl)-4-iodo-benzenesulfonamide (223) by means of the procedure described in Patent Application No. WO 01/38322 A1 by means of 3,4-dimethoxy-aniline with 4- Reaction with iodine-benzenesulfonyl chloride gave the title compound 223 (yield: 80%). LRMS: 419.2 (calc.); 420.2 (Get Value) (MH) +.
步驟2：N－(2－胺基－苯基)－4－(3,4－二甲氧基－苯基胺磺醯基)－苯甲醯胺(224) 將223(705 mg，1.7 mmol)、1,2－苯二胺(199 mg，1.84 mmol)、Pd(OAc)2 (0.25 mmol，15%)及1,1'－雙(二苯膦基)二茂鐵(160 mg，0.29 mmol)之混合物懸浮於脫氣DMF(10 mL)中，以Et3 N(700μL，5.04 mmol)進行處理，在70℃下CO氣氛(氣球)下將其加熱18小時。真空下蒸發DMF後由急驟層析法(溶離劑AcOEt：己烷，3：1)純化殘餘物產生標題化合物224(100 mg,產率14%)。1 H－NMR(CD3 OD－d4 ),δ(ppm)：10.05(s,1H),9.76(s,1H),8.06(d,J＝8.3 Hz,2H),7.79(d,J＝7.8 Hz,2H),7.11(bs,1H),6.94(bs,1H),6.77－6.69(m,3H),6.54(bs,2H),4.91(bs,2H),3.62(s,3H)。 Step 2: N-(2-Amino-phenyl)-4-(3,4-dimethoxy-phenylamine sulfonyl)-benzamide (224) 223 (705 mg, 1.7 mmol ), 1,2-phenylenediamine (199 mg, 1.84 mmol), Pd(OAc) 2 (0.25 mmol, 15%) and 1,1'-bis(diphenylphosphino)ferrocene (160 mg, 0.29) mmol) of the mixture was suspended in degassed DMF (10 mL) in order to Et 3 N (700μL, 5.04 mmol ) treated at 70 deg.] C under CO atmosphere (balloon) was heated for 18 hours. The residue was purified by flash chromatography eluting elut elut elut elut elut elut elut elut 1 H-NMR (CD 3 OD-d 4 ), δ (ppm): 10.05 (s, 1H), 9.76 (s, 1H), 8.06 (d, J = 8.3 Hz, 2H), 7.79 (d, J = 7.8 Hz, 2H), 7.11 (bs, 1H), 6.94 (bs, 1H), 6.77-6.69 (m, 3H), 6.54 (bs, 2H), 4.91 (bs, 2H), 3.62 (s, 3H).
N－(2－胺基－苯基)－4－[4－(4－甲氧基－苯基)－嘧啶－2－基硫基甲基]－苯甲醯胺(228a)步驟1：4－[4－(4－甲氧基－苯基)－嘧啶－2－基硫基甲基]－苯甲酸甲酯(226a) 向4－(4－甲氧基－苯基)－嘧啶－2－硫醇(225a)(1.00 g，4.58 mmol)之DMF(30 mL)溶液中加入4－溴甲基－苯甲酸甲酯(1.05 g，4.58 mmol)。將該混合物在60℃下加熱1小時且蒸乾形成化合物226a，該化合物未經純化即用於下一步驟。LRMS＝366.4(計算值)，367.4(實驗值)。 N-(2-Amino-phenyl)-4-[4-(4-methoxy-phenyl)-pyrimidin-2-ylthiomethyl]-benzamide (228a) Step 1:4 -[4-(4-Methoxy-phenyl)-pyrimidin-2-ylthiomethyl]-benzoic acid methyl ester (226a) to 4-(4-methoxy-phenyl)-pyrimidine-2 -Methyl 4-bromomethyl-benzoate (1.05 g, 4.58 mmol) was added to a solution of EtOAc (EtOAc (EtOAc). The mixture was heated at 60 <0>C for 1 h and evaporated to dryness to afford compound 226a. LRMS = 366.4 (calculated), 367.4 (exp.).
步驟2：4－[4－(4－甲氧基－苯基)－嘧啶－2－基硫基甲基]]－苯甲酸(227a) 在室溫下向THF(20 ml)及MeOH(20 ml)中之226a(4.58 mmol)經攪拌溶液中加入LiOH－H2 O(960 mg，22.9 mmol)水(50 ml)溶液。在室溫下攪拌該反應混合物18小時，以水稀釋且以1 N HCl(pH 5－6)酸化形成沉澱物，過濾收集該沉澱物，將其以水洗滌及乾燥以產生標題化合物227a(1.64 g，產率99%)。LRMS(計算值)：352.4，(實驗值)：353.4。 Step 2: 4-[4-(4-Methoxy-phenyl)-pyrimidin-2-ylthiomethyl]]-benzoic acid (227a) THF (20 mL) and MeOH (20) A solution of LiOH-H 2 O (960 mg, 22.9 mmol) in water (50 ml) was added to a stirred solution of 226a (4.58 mmol). The reaction mixture was stirred at room temperature for 18 hours, diluted with water and EtOAc (EtOAc) (EtOAc) g, yield 99%). LRMS (calculated): 352.4, (exp.): 353.4.
步驟3：N－(2－胺基－苯基)－4－[4－(4－甲氧基－苯基)－嘧啶－2－基硫基甲基]－苯甲醯胺(228a) 依照流程1步驟5(實例1)所述之程序藉由使酸227a與1,2－苯二胺偶合獲得標題化合物228a(產率80%)。1 H NMR：(DMSO)δ(ppm)：9.57(bs,1H),8.59(d,J＝5.5 Hz,2H),8.16(d,J＝7.0 Hz,2H),7.88(d,J＝8.2 Hz,2H),7.70(d,J＝5.0 Hz,1H),7.57(d,J＝8.2,2H),7.12－7.07(m,2H),6.93(dd,J＝8.2,7.0 Hz,1H),6.73(dd,J＝8.2,1.6 Hz,1H),6.55(dt,J＝8.6,1.1 Hz,1H),4.86(bs,2H),4.55(s,2H),3.83(s,3H)LRMS：(計算值)442.5；(獲得值)443.5(MH)＋ 。 Step 3: N-(2-Amino-phenyl)-4-[4-(4-methoxy-phenyl)-pyrimidin-2-ylthiomethyl]-benzamide (228a) according to The procedure described in Scheme 1, Step 5 (Example 1) was obtained by coupling the acid 227a with 1,2-phenylenediamine to give the title compound 228a (yield 80%). 1 H NMR: (DMSO) δ (ppm): 9.57 (bs, 1H), 8.59 (d, J = 5.5 Hz, 2H), 8.16 (d, J = 7.0 Hz, 2H), 7.88 (d, J = 8.2 Hz, 2H), 7.70 (d, J = 5.0 Hz, 1H), 7.57 (d, J = 8.2, 2H), 7.12-7.07 (m, 2H), 6.93 (dd, J = 8.2, 7.0 Hz, 1H) , 6.73 (dd, J = 8.2, 1.6 Hz, 1H), 6.55 (dt, J = 8.6, 1.1 Hz, 1H), 4.86 (bs, 2H), 4.55 (s, 2H), 3.83 (s, 3H) LRMS : (calculated value) 442.5; (obtained value) 443.5 (MH) + .
用與化合物228a實例121(流程47，表1)所述相同之程序自硫酚225b－f開始經中間體226b－f及227b－f(流程47)製備實例121－125(化合物228b－f)。Examples 121-125 (compounds 228b-f) were prepared from thiophenol 225b-f starting from thiophenol 225b-f using intermediates 226b-f and 227b-f (Scheme 47) using the same procedure as described for Compound 228a, Example 121 (Scheme 47, Table 1). .
4－[4－(2－胺基－苯胺甲醯基)－苄胺基]－2－甲基硫基－嘧啶－5－甲酸醯胺(232a)步驟1：4－[(5－氰基－2－甲基硫基－嘧啶－4－基胺基)－甲基]－苯甲酸甲酯(230a) 在室溫下向4－胺基－2－甲基硫基－嘧啶－5－腈(229a)(200 mg，1.2 mmol)之DME(10 ml)溶液中加入4－溴甲基－苯甲酸甲酯(274 mg，1.2 mmol)及K2 CO3 (663 mg，4.8 mmol)。將反應混合物在100℃下加熱5小時，在60℃下隔夜，將其冷卻，過濾且在真空下濃縮。該粗產物未經進一步純化即用於下一步反應中。LRMS：314.3(計算值)；315.3(獲得值)(MH)＋ 。 4-[4-(2-Amino-anilinemethanyl)-benzylamino]-2-methylsulfanyl-pyrimidine-5-carboxylic acid decylamine (232a) Step 1: 4-[(5-Cyano) 4-methylthio-pyrimidin-4-ylamino)-methyl]-benzoic acid methyl ester (230a) to 4-amino-2-methylthio-pyrimidine-5-carbonitrile at room temperature (229a) (200 mg, 1.2 mmol) of DME (10 ml) was added a solution of 4-bromomethyl - benzoic acid methyl ester (274 mg, 1.2 mmol) and K 2 CO 3 (663 mg, 4.8 mmol). The reaction mixture was heated at 100 <0>C for 5 h, cooled overnight at 60 <0>C, filtered and concentrated in vacuo. This crude product was used in the next reaction without further purification. LRMS: 314.3 (calculated); 315.3 (obtained) (MH) + .
步驟2：4－[(5－胺甲醯基－2－甲基硫基－嘧啶－4－基胺基)－甲基]－苯甲酸(231a) 依照實例121步驟2(流程47)所述之程序但以化合物230a代替化合物226a獲得標題化合物231a(227 mg，產率60%)。LRMS(計算值)：318.3，(實驗值)：319.3。 Step 2: 4-[(5-Aminocarbamido-2-methylsulfanyl-pyrimidin-4-ylamino)-methyl]-benzoic acid (231a) as described in Example 121 Step 2 (Scheme 47) The title compound 231a (227 mg, yield 60%) was obtained from Compound 230a. LRMS (calculated): 318.3, (exp.): 319.3.
步驟3：4－[4－(2－胺基－苯胺甲醯基)－苄胺基]－2－甲基硫基－嘧啶－5－甲酸醯胺(232a) 依照流程1步驟5(實例1)所述之程序藉由酸231a與1,2－苯二胺之偶合反應獲得標題化合物232a(產率80%)。1 H NMR：(DMSO)δ(ppm)：9.39(bs,1H),9.35(bs,2H),8.32(s,1H),7.71(d,J＝8.4 Hz,2H),7.22(d,J＝8.2 Hz,2H),6.94(d,J＝7.6 Hz,1H),6.75(dt,J＝1.4,8.2,1H),6.56(dd,J＝1.5,8.0 Hz,1H),6.39(t,J＝7.4 Hz,1H),4.54(s,2H),2.30(s,3H)。LRMS：(計算值)408.5；(獲得值)409.5(MH)＋ 。 Step 3: 4-[4-(2-Amino-anilinemethanyl)-benzylamino]-2-methylsulfanyl-pyrimidine-5-carboxylic acid decylamine (232a) according to Scheme 1 Step 5 (Example 1 The procedure described above was carried out by coupling of acid 231a with 1,2-phenylenediamine to give the title compound 232a (yield: 80%). 1 H NMR: (DMSO) δ (ppm): 9.39 (bs, 1H), 9.35 (bs, 2H), 8.32 (s, 1H), 7.71 (d, J = 8.4 Hz, 2H), 7.22 (d, J) = 8.2 Hz, 2H), 6.94 (d, J = 7.6 Hz, 1H), 6.75 (dt, J = 1.4, 8.2, 1H), 6.56 (dd, J = 1.5, 8.0 Hz, 1H), 6.39 (t, J = 7.4 Hz, 1H), 4.54 (s, 2H), 2.30 (s, 3H). LRMS: (calculated) 408.5; (obtained) 409.5 (MH) + .
4－[4－(2－胺基－苯胺甲醯基)－苄胺基]－2－吡啶－3－基－嘧啶－5－甲酸醯胺(232b) 依照與用於化合物232a實例126(流程48)所述相同之程序自胺基腈229b開始經由中間體230b及231b獲得標題化合物232b。1 H NMR：(DMSO－d6 )δ(ppm)：9.48(bs,3H),8.86(s,1H),8.74－8.73(m,1H),8.64－8.61(m,1H),8.19(bs,1H),7.98(d,J＝8.8 Hz,2H),7.71(bs,1H),7.60－7.57(m,3H),7.18(d,J＝8.4 Hz,1H),7.00(t,J＝6.8 Hz,1H),6.81(d,J＝8.8 Hz,1H),6.63(t,J＝8.0 Hz,1H),4.95(d,J＝6.8 Hz,2H)。 4-[4-(2-Amino-anilinemethanyl)-benzylamino]-2-pyridin-3-yl-pyrimidine-5-carboxylic acid decylamine (232b) was used according to Example 126 for compound 232a (flow 48) The same procedure was carried out starting from the amino nitrile 229b to give the title compound 232b via intermediates 230b and 231b. 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.48 (bs, 3H), 8.86 (s, 1H), 8.74 - 8.73 (m, 1H), 8.64 - 8.61 (m, 1H), 8. , 1H), 7.98 (d, J = 8.8 Hz, 2H), 7.71 (bs, 1H), 7.60-7.57 (m, 3H), 7.18 (d, J = 8.4 Hz, 1H), 7.00 (t, J = 6.8 Hz, 1H), 6.81 (d, J = 8.8 Hz, 1H), 6.63 (t, J = 8.0 Hz, 1H), 4.95 (d, J = 6.8 Hz, 2H).
N－(2－胺基苯基)－4－(4－(5－(三氟甲基)吡啶－2－基)六氫吡嗪－1－基)苯甲醯胺(237)步驟1：4－[4－(4－三氟甲基－吡啶－2－基)－六氫吡嗪－1－基]－苯甲酸第三丁酯(235) 向1－(4－三氟甲基－吡啶－2－基)－六氫吡嗪(233)(500 mg，2.16 mmol)之DMSO溶液中加入4－氟－苯甲酸第三丁酯(466 mg，2.37 mmol)(234)及K2 CO3 (1.2 g，11.3 mmol)。在130℃下加熱該混合物16小時，將其冷卻，過濾且在真空下濃縮之。由急驟層析法(溶離劑AcOEt－己烷40：60至純AcOEt)純化該殘餘物產生標題化合物244(162 mg，產率18%)。LRMS：(計算值)406.4：(實驗值)407.4(MH)＋ 。 N-(2-Aminophenyl)-4-(4-(5-(trifluoromethyl)pyridin-2-yl)hexahydropyrazin-1-yl)benzamide (237) Step 1: 3-[4-(4-Trifluoromethyl-pyridin-2-yl)-hexahydropyrazin-1-yl]-benzoic acid tert-butyl ester (235) to 1-(4-trifluoromethyl- Add 4-fluoro-benzoic acid tert-butyl ester (466 mg, 2.37 mmol) (234) and K 2 CO to a solution of pyridin-2-yl)-hexahydropyrazine (233) (500 mg, 2.16 mmol) in DMSO 3 (1.2 g, 11.3 mmol). The mixture was heated at 130 ° C for 16 hours, cooled, filtered and concentrated in vacuo. The residue was purified by flash chromatography eluting elut elut elut elut elut elut LRMS: (calculated) 406.4: (found) 407.4 (MH) + .
步驟2：4－[4－(4－三氟甲基－吡啶－2－基)－六氫吡嗪－1－基]－苯甲酸(236) 依照流程28步驟5(實例68)所述之程序自化合物235開始獲得標題化合物236(產率99%)。LRMS 350.3(計算值)，351.3(實驗值)。 Step 2: 4-[4-(4-Trifluoromethyl-pyridin-2-yl)-hexahydropyrazin-1-yl]-benzoic acid (236) according to Step 28 (Example 68) of Scheme 28 The title compound 236 was obtained starting from compound 235 (yield: 99%). LRMS 350.3 (calculated), 351.3 (exp.).
步驟3：N－(2－胺基－苯基)－4－[4－(4－三氟甲基－吡啶－2－基)－六氫吡嗪－1－基]－苯甲醯胺(237) 依照流程1步驟5(實例1)所述之程序藉由使酸236與1,2－苯二胺偶合獲得標題化合物237(產率96%)。1 H NMR：(DMSO)δ(ppm)：9.43(bs,1H),8.43(s,1H),7.93(bs,2H),7.88(d,J＝8.8 Hz,2H),7.81(dd,J＝2.4,8.8 Hz,1H),7.13(d,J＝8.0 Hz,1H),7.04－7.00(m,3H),6.96(t,J＝7.6 Hz,1H),6.76(d,J＝8.0 Hz,1H),6.58(t,J＝7.6 Hz,1H),4.84(bs,2H),3.82－3.79(m,4H),3.44－3.40(m,4H)。LRMS：(計算值)440.4；(實驗值)441.4(MH)＋ 。 Step 3: N-(2-Amino-phenyl)-4-[4-(4-trifluoromethyl-pyridin-2-yl)-hexahydropyrazin-1-yl]-benzamide ( 237) The title compound 237 (yield: 96%) was obtained by coupling of acid 236 with 1,2-phenylenediamine according to the procedure of procedure 1 (5). 1 H NMR: (DMSO) δ (ppm): 9.43 (bs, 1H), 8.43 (s, 1H), 7.93 (bs, 2H), 7.88 (d, J = 8.8 Hz, 2H), 7.81 (dd, J =2.4, 8.8 Hz, 1H), 7.13 (d, J = 8.0 Hz, 1H), 7.04-7.00 (m, 3H), 6.96 (t, J = 7.6 Hz, 1H), 6.76 (d, J = 8.0 Hz) , 1H), 6.58 (t, J = 7.6 Hz, 1H), 4.84 (bs, 2H), 3.82-3.79 (m, 4H), 3.44 - 3.40 (m, 4H). LRMS: (calculated) 440.4; (found) 441.4 (MH) + .
N－(2－胺基－苯基)－4－({3－[3－(1,3－二酮基－1,3－二氫－異吲哚－2－基)－丙氧基]－4－甲氧基－苯胺基}－甲基)－苯甲醯胺(239)步驟1：N－(2－胺基－苯基)－4－({3－[3－(1,3－二酮基－1,3－二氫－異吲哚－2－基)－丙氧基]－4－甲氧基－苯胺基}－甲基)－苯甲醯胺(239) 在室溫下向N－(2－胺基－苯基)－4－[(3－羥基－4－甲氧基－苯胺基)－甲基]－苯甲醯胺(212e)(586 mg，0.66 mmol)之DMF(10 ml)溶液中加入2－(3－溴－丙基)－異吲哚－1,3－二酮(238)(176 mg，0.66 mmol)及K2 CO3 (365 mg，2.64 mmol)。將反應混合物在100℃下加熱1小時接著在60℃下加熱隔夜，使其冷卻，經過濾且在真空下濃縮之。由急驟層析法(溶離劑由AcOEt－己烷(40：60)至純AcOEt)純化該殘餘物產生239(168 mg，產率46%)。1 H NMR：(DMSO)δ(ppm)：9.57(bs,1H),7.88(d,J＝8.0 Hz,2H),7.84－7.75(m,4H),7.44(d,J＝8.0 Hz,2H),7.12(d,J＝7.2 Hz,1H),6.92(t,J＝8.8 Hz,1H),6.75(dd,J＝1.2,7.6 Hz,1H),6.57(d,J＝8.4 Hz,2H),6.27(d,J＝2.4 Hz,1H),6.00－5.93(m,2H),4.87(s,2H),4.27(d,J＝6.0 Hz,2H),3.89(dd,J＝5.6,6.0 Hz,2H),3.74(dd,J＝6.4,6.8 Hz,2H),3.42(s,3H),2.06－2.01(m,2H)。LRMS：(計算植)550.4；(實驗植)551.5(MH)＋ 。 N-(2-Amino-phenyl)-4-({3-[3-(1,3-diketo-1,3-1,3-dihydro-isoindol-2-yl)-propoxy] 4-methoxy-anilino}-methyl)-benzamide (239) Step 1: N-(2-Amino-phenyl)-4-({3-[3-(1,3) -diketo-1,3-dihydro-isoindol-2-yl)-propoxy]-4-methoxy-anilino}-methyl)-benzamide (239) at room temperature Downward N-(2-amino-phenyl)-4-[(3-hydroxy-4-methoxy-anilino)-methyl]-benzamide (212e) (586 mg, 0.66 mmol) 2-(3-Bromo-propyl)-isoindole-1,3-dione (238) (176 mg, 0.66 mmol) and K 2 CO 3 (365 mg, 2.64) were added to a solution of DMF (10 ml) Mm). The reaction mixture was heated at 100 <0>C for 1 h then heated at 60 <0>C overnight, cooled, filtered and concentrated in vacuo. The residue was purified by flash chromatography (EtOAc: EtOAc (EtOAc:EtOAc) 1 H NMR: (DMSO) δ (ppm): 9.57 (bs, 1H), 7.88 (d, J = 8.0 Hz, 2H), 7.84-7.75 (m, 4H), 7.44 (d, J = 8.0 Hz, 2H ), 7.12 (d, J = 7.2 Hz, 1H), 6.92 (t, J = 8.8 Hz, 1H), 6.75 (dd, J = 1.2, 7.6 Hz, 1H), 6.57 (d, J = 8.4 Hz, 2H) ), 6.27 (d, J = 2.4 Hz, 1H), 6.00-5.93 (m, 2H), 4.87 (s, 2H), 4.27 (d, J = 6.0 Hz, 2H), 3.89 (dd, J = 5.6, 6.0 Hz, 2H), 3.74 (dd, J = 6.4, 6.8 Hz, 2H), 3.42 (s, 3H), 2.06-2.01 (m, 2H). LRMS: (calculated plant) 550.4; (experimental plant) 551.5 (MH) + .
4－[1－(6－乙醯基－苯幷[1,3]二氧雜環戊烷－5－基胺基)－乙基]－N－(2－胺基－苯基)－苯甲醯胺(240) 依照與用於化合物212a實例99(流程42，表1)所述相同之程序，自1－(6－胺基－苯幷[1,3]間二氧雜環戊烷－5－基)－乙酮及4－乙醯基苯甲酸(流程51)開始獲得標題化合物240。1 H NMR：(DMSO－d6 )δ(ppm)：8.69(s,1H),7.03(d,J＝7.8 Hz,2H),6.64(d,J＝7.8 Hz,2H),6.29(dd,J＝8.3,7.8 Hz,1H),6.09(t,J＝7.8,7.3 Hz,1H),5.90(d,J＝7.8,1H),5.72(d,J＝6.8 Hz,1H),5.70(s,1H),5.16(d,J＝8.8 Hz,1H),4.88(s,1H),4.84(s,1H),4.07(bd,1H),2.30(s,3H),0.62(d,J＝6.83,3H)。 4-[1-(6-Ethylidene-benzoquinone[1,3]dioxol-5-ylamino)-ethyl]-N-(2-amino-phenyl)-benzene Formamide (240) was obtained from 1-(6-amino-benzoquinone [1,3] dioxolane according to the same procedure as described for the compound 212a, Example 99 (Scheme 42, Table 1). -5-yl)-ethanone and 4-ethylmercaptobenzoic acid (Scheme 51). 1 H NMR: (DMSO-d 6 ) δ (ppm): 8.69 (s, 1H), 7.03 (d, J = 7.8 Hz, 2H), 6.64 (d, J = 7.8 Hz, 2H), 6.29 (dd, J = 8.3, 7.8 Hz, 1H), 6.09 (t, J = 7.8, 7.3 Hz, 1H), 5.90 (d, J = 7.8, 1H), 5.72 (d, J = 6.8 Hz, 1H), 5.70 (s) , 1H), 5.16 (d, J = 8.8 Hz, 1H), 4.88 (s, 1H), 4.84 (s, 1H), 4.07 (bd, 1H), 2.30 (s, 3H), 0.62 (d, J = 6.83, 3H).
N－(2－胺基－苯基)－4－{[4,5－二甲氧基－2－(3,4,5－三甲氧基－苯甲醯基)－苯胺基]－甲基}－苯甲醯胺(245)步驟1：(4,5－二甲氧基－2－硝基－苯基)－(3,4,5－三甲氧基－苯基)－甲醇(241)： 在N2 氣氛下向－經烘乾之圓底燒瓶中裝入5－碘－1,2,3－三甲氧基苯(2.92 g，9.93 mmol)且加入THF(31 mL)。將溶液冷卻至－78℃且逐滴加入1.5 M t－BuLi之戊烷溶液(13.6 mL，20.57 mmol)。攪拌該混合物1小時且在N2 氣氛下經由套管將其轉移至6－硝基藜蘆醛(2.02 g，9.57 mmol)之THF(12 mL)預***液(－78℃)中。攪拌所得混合物2小時且將其緩緩溫至0℃，以NH4 Cl飽和水溶液中止且使其溫至室溫。在真空下移除溶劑且使殘餘物在水與DCM之間分溶。收集有機層且以鹽水洗滌，將其經Na2 SO4 乾燥且在真空下濃縮之。使用EtOAc/DCM(9：91)由急驟層析法純化該殘餘物產生標題化合物241(1.46 g，產率40%)。1 H NMR(CDCl3 )δ(ppm)：7.61(s,1H),7.16(s,1H),6.58(s,2H),6.45(s,1H),6.09(s,1H),3.97(s,3H),3.93(s,3H),3.84(s,3H),3.83(s,6H)。m/z：402.4(MH＋ )。 N-(2-Amino-phenyl)-4-{[4,5-dimethoxy-2-(3,4,5-trimethoxy-benzylidene)-anilino]-methyl }-benzamide (245) Step 1: (4,5-Dimethoxy-2-nitro-phenyl)-(3,4,5-trimethoxy-phenyl)-methanol (241) : under an N 2 atmosphere - dried round-bottomed flask was charged with the 5-iodo-1,2,3-methoxybenzene (2.92 g, 9.93 mmol) was added and THF (31 mL). The solution was cooled to -78 ° C and a solution of 1.5 M t-BuLi in pentane (13.6 mL, 20.57 mmol). The mixture was stirred for 1 hour under an atmosphere of N 2 and via cannula transferred to a 6-nitro-veratraldehyde (2.02 g, 9.57 mmol) of THF (12 mL) precooled solution (-78 deg.] C) in. The resulting mixture was stirred for 2 h and slowly warmed to 0 ° C, quenched with saturated aqueous NH 4 Cl and warmed to room temperature. The solvent was removed under vacuum and the residue was partitioned between water and DCM. The organic layer was collected and washed with brine, which was dried over Na 2 SO 4 and concentrated under a vacuum. The residue was purified by flash chromatography eluting elut elut elut elut elut elut 1 H NMR (CDCl 3 ) δ (ppm): 7.61 (s, 1H), 7.16 (s, 1H), 6.58 (s, 2H), 6.45 (s, 1H), 6.09 (s, 1H), 3.97 (s) , 3H), 3.93 (s, 3H), 3.84 (s, 3H), 3.83 (s, 6H). m/z: 402.4 (MH + ).
步驟2：(4,5－二甲氧基－2－硝基－苯基)－(3,4,5－三甲氧基－苯基)－甲酮(242)： 在0℃下將粉末4分子篩(583 mg)及重鉻酸吡錠(2.17 g，5.77 mmol)相繼加入至中間體241(1.46 g，3.84 mmol)之無水DCM(38.5 mL)溶液中。在室溫下攪拌該混合物15小時。加入更多PDC(290 mg,0.770 mmol)且再攪拌該混合物4小時，將其以醚稀釋且經矽藻土墊過濾。濃縮濾液且由急驟層析法使用EtOAc/DCM(7：93)純化褐色固體產生呈黃色固體之標題化合物242(551mg，41%)。1 H NMR(CDCl3 )δ(ppm)：7.72(s,1H),6.99(s,2H),6.86(s,1H),4.06(s,3H),4.00(s,3H),3.93(s,3H),3.84(s,6H)。m/z：378.4(MH＋ )。 Step 2: (4,5-Dimethoxy-2-nitro-phenyl)-(3,4,5-trimethoxy-phenyl)-methanone (242): powder 4 at 0 ° C Molecular sieves (583 mg) and dichromate pyridinium (2.17 g, 5.77 mmol) were added sequentially to a solution of intermediate 241 (1.46 g, 3.84 mmol) in anhydrous DCM (38.5 mL). The mixture was stirred at room temperature for 15 hours. More PDC (290 mg, 0.770 mmol) was added and the mixture was stirred further for 4 h then diluted with ether and filtered thru a pad. The filtrate was concentrated and EtOAc EtOAc EtOAcjjjjjj 1 H NMR (CDCl 3 ) δ (ppm): 7.72 (s, 1H), 6.99 (s, 2H), 6.86 (s, 1H), 4.06 (s, 3H), 4.00 (s, 3H), 3.93 (s) , 3H), 3.84 (s, 6H). m/z: 378.4 (MH + ).
步驟3：(2－胺基－4,5－二甲氧基－苯基)－(3,4,5－三甲氧基－苯基)－甲酮(243)： 將鐵粉(653 mg，11.7 mmol)加入至溶於EtOH(5.11 mL)、H2 O(2.56 mL)及AcOH(5.11 mL)之混合物中的中間體199(552 mg，1.46 mmol)懸浮液中且向該溶液中加入2滴濃HCl。劇烈攪拌該混合物同時回流1小時，將其冷卻至室溫且經矽藻土墊過濾。在真空下濃縮濾液且使該含水殘餘物在DCM與H2 O之間分溶。以飽和NaHCO3 洗滌該有機層，將其經Na2 SO4 乾燥且在真空下濃縮產生標題化合物243(393 mg，77%)。1 H NMR(CDCl3 )δ(ppm)：7.00(s,1H),6.88(s,2H),6.23(s,1H),3.92(m,6H),3.88(s,6H),3.70(s,3H)。m/z：348.4(MH＋ )。 Step 3: (2-Amino-4,5-dimethoxy-phenyl)-(3,4,5-trimethoxy-phenyl)-methanone (243): Iron powder (653 mg, 11.7 mmol) was added to a suspension of intermediate 199 (552 mg, 1.46 mmol) in a mixture of EtOH (5.11 mL), H 2 O (2.56 mL) and AcOH (5.11 mL) and added to the solution 2 Concentrated HCl. The mixture was stirred vigorously while refluxing for 1 hour, cooled to room temperature and filtered through a pad of Celite. The filtrate was concentrated and the aqueous residue was partitioned between DCM and H 2 O in vacuo. Washed the organic layer with saturated NaHCO 3, which was 2 SO 4 dried and concentrated in vacuo to give the title compound 243 (393 mg, 77%) over Na. 1 H NMR (CDCl 3 ) δ (ppm): 7.00 (s, 1H), 6.88 (s, 2H), 6.23 (s, 1H), 3.92 (m, 6H), 3.88 (s, 6H), 3.70 (s) , 3H). m/z: 348.4 (MH + ).
步驟4：4－{[4,5－二甲氧基－2－(3,4,5－三甲氧基－苯甲醯基)－苯胺基]－甲基}－苯甲酸(244) 依照與流程3步驟2(實例12)對還原胺化反應所述相同之程序自化合物243開始獲得標題化合物244(產率46%)，m/z：482.5(MH＋ )。 Step 4: 4-{[4,5-Dimethoxy-2-(3,4,5-trimethoxy-benzylidenyl)-anilino]-methyl}-benzoic acid (244) according to The title compound 244 (yield 46%), m/z: 482.5 (MH + ).
步驟5：N－(2－胺基－苯基)4－{[4,5－二甲氧基－2－(3,4,5－三甲氧基－苯甲醯基)－苯胺基]－甲基}－苯甲醯胺(245) 依照與流程1步驟5(實例1)對BOP偶合反應所述相同之程序用化合物244作為原料獲得標題化合物245(產率38%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.61(s,1H),9.24(t,J＝5.7 Hz,1H),7.95(d,J＝8.2 Hz,2H),7.51(d,J＝8.2 Hz,2H)7.13(d,J＝8.0 Hz,1H),6.98(s,1H),6.94(td,J＝7.6,1.2 Hz,1H)6.84(s,2H),6.75(dd,J＝8.0,1.3 Hz,1H),6.57(t,J＝7.2 Hz,1H),6.35(s,1H),4.89(s,2H),4.62(d,J＝5.7 Hz,2H),3.79(s,6H),3.76(s,3H),3.73(s,3H),3.54(s,3H)。m/z：572.5(MH＋ )。 Step 5: N-(2-Amino-phenyl)4-{[4,5-dimethoxy-2-(3,4,5-trimethoxy-benzomethyl)-anilino]- Methyl}-benzamide (245) The title compound 245 (yield: 38%) was obtained from the compound 244 as the starting material. 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.61 (s, 1H), 9.24 (t, J = 5.7 Hz, 1H), 7.95 (d, J = 8.2 Hz, 2H), 7.51 (d, J = 8.2 Hz, 2H) 7.13 (d, J = 8.0 Hz, 1H), 6.98 (s, 1H), 6.94 (td, J = 7.6, 1.2 Hz, 1H) 6.84 (s, 2H), 6.75 (dd, J = 8.0, 1.3 Hz, 1H), 6.57 (t, J = 7.2 Hz, 1H), 6.35 (s, 1H), 4.89 (s, 2H), 4.62 (d, J = 5.7 Hz, 2H), 3.79 ( s, 6H), 3.76 (s, 3H), 3.73 (s, 3H), 3.54 (s, 3H). m/z: 572.5 (MH + ).
N－(2－胺基－苯基)－4－{[4,5－二甲氧基－2－(4－甲氧基－苯甲醯基)－苯胺基]－甲基}－苯甲醯胺(246) 依照與實例131所述相同之程序但以市售格林納(Grignard)試劑4－甲氧苯基溴化鎂代替自5－碘－1,2,3－三甲氧基苯及正丁基鋰獲得之有機鋰試劑，獲得標題化合物246(總產率8.4%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.62(s,1H),9.10(t,J＝5.7 Hz,1H),7.95(d,J＝8.2,2H),7.57(d,J＝8.8 Hz,2H),7.52(d,J＝8.2 Hz,2H),7.14(d,J＝6.6 Hz,1H),7.04(d,J＝8.8 Hz,2H),6.95(td,J＝8.2,1.6 Hz,2H),6.94(s,1H),6.76(dd,J＝7.8,1.4 Hz,1H),6.58(t,J＝6.5 Hz,2H),6.35(s,2H),4.90(s,2H),4.62(d,J＝5.3 Hz,2H),3.84(s,3H),3.76(s,3H),3.54(s,3H)。m/z：512.6(MH＋ )。 N-(2-Amino-phenyl)-4-{[4,5-dimethoxy-2-(4-methoxy-benzylidenyl)-anilino]-methyl}-benzene Indoleamine (246) was replaced by the same procedure as described in Example 131 except that the commercially available Grignard reagent 4-methoxyphenylmagnesium bromide was substituted for 5-iodo-1,2,3-trimethoxybenzene and The organolithium reagent obtained from n-butyllithium gave the title compound 246 (yield 8.4%). 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.62 (s, 1H), 9.10 (t, J = 5.7 Hz, 1H), 7.95 (d, J = 8.2, 2H), 7.57 (d, J) = 8.8 Hz, 2H), 7.52 (d, J = 8.2 Hz, 2H), 7.14 (d, J = 6.6 Hz, 1H), 7.04 (d, J = 8.8 Hz, 2H), 6.95 (td, J = 8.2 , 1.6 Hz, 2H), 6.94 (s, 1H), 6.76 (dd, J = 7.8, 1.4 Hz, 1H), 6.58 (t, J = 6.5 Hz, 2H), 6.35 (s, 2H), 4.90 (s , 2H), 4.62 (d, J = 5.3 Hz, 2H), 3.84 (s, 3H), 3.76 (s, 3H), 3.54 (s, 3H). m/z: 512.6 (MH + ).
N－(2－胺基－苯基)－4－[(1－甲基－1H－苯幷咪唑－5－基胺基)－甲基]－苯甲醯胺(252)步驟1：N1－甲基－4－硝基－苯－1,2－二胺(248)： 以40% w/w MeNH2 之水溶液(10 mL，128 mmol)處理氟化物247(5.41 g，34.7 mmol)之DMF(40 mL)溶液。在90℃下攪拌該混合物3小時，將其以EtOAc稀釋，以NaHCO3 飽和水溶液洗滌。將有機相經MgSO4 乾燥，蒸發且由急驟層析法(溶離劑在CH2 Cl2 中50% EtOAc)純化殘餘物產生化合物248(5.31 g，產率92%)。1 H NMR：(CDCl3 )δ(ppm)：7.75(dd,J＝2.6,8.8 Hz,1H),7.53(d,J＝2.6 Hz,1H),6.45(d,J＝8.8 Hz,1H),4.24(bs,3H),2.91(s,3H)。LRMS：(計算值)167.2；(實驗值)168.1(MH)＋ 。 N-(2-Amino-phenyl)-4-[(1-methyl-1H-benzoimidazole-5-ylamino)-methyl]-benzamide (252) Step 1: N1- Methyl-4-nitro-benzene-1,2-diamine (248): Treatment of fluoride 247 (5.41 g, 34.7 mmol) of DMF in 40% w/w MeNH 2 (10 mL, 128 mmol) (40 mL) solution. The mixture was stirred for 3 hours at 90 ℃, it was diluted with EtOAc, washed with saturated aqueous NaHCO 3. The organic phase was dried MgSO 4, evaporated and (eluent CH 2 Cl 2 in the 50% EtOAc) produced by The residue was purified by flash chromatography, compound 248 (5.31 g, yield 92%). 1 H NMR: (CDCl 3 ) δ (ppm): 7.75 (dd, J = 2.6, 8.8 Hz, 1H), 7.53 (d, J = 2.6 Hz, 1H), 6.45 (d, J = 8.8 Hz, 1H) , 4.24 (bs, 3H), 2.91 (s, 3H). LRMS: (calculated) 167.2; (experimental) 168.1 (MH) + .
步驟2.1－甲基－5－硝基－1H－苯幷咪唑(249) 向二胺248(1.14 g，6.80 mmol)之CH2 Cl2 (10 mL)懸浮液中加入原甲酸三甲酯(5 mL，46 mmol，6.7當量)(或任意其它所選醯化劑，6當量)隨後加入TFA(0.43 mL，5.6 mmol，0.8當量)，且在室溫下攪拌該混合物2小時。過濾收集該沉澱物，將其以CH2 Cl2 洗滌及乾燥產生呈TFA鹽之標題化合物249(1.23 g，產率62%)。1 H NMR：(CDCl3 )δ(ppm)：8.57(d,J＝1.8 Hz,1H),8.20(dd,J＝1.8,9.2 Hz,1H),8.18(s,1H),7.54(d,J＝9.2 Hz,1H),3.94(s,3H)。LRMS：(計算值)177.2；(獲得值)178.1(MH)＋ 。 Step 2. 1-methyl-5-nitro-benzene Bing -1H- imidazole (249) diamine 248 (1.14 g, 6.80 mmol) was added of to the CH 2 Cl 2 (10 mL) and trimethyl orthoformate (5 </ RTI><RTIgt;</RTI><RTIgt;</RTI><RTIgt;</RTI></RTI><RTIgt;</RTI></RTI></RTI><RTIgt; The precipitate was collected by filtration, which was washed with CH 2 Cl 2 and dried to give the TFA salt of the title compound 249 (1.23 g, yield 62%). 1 H NMR: (CDCl 3 ) δ (ppm): 8.57 (d, J = 1.8 Hz, 1H), 8.20 (dd, J = 1.8, 9.2 Hz, 1H), 8.18 (s, 1H), 7.54 (d, J = 9.2 Hz, 1H), 3.94 (s, 3H). LRMS: (calculated) 177.2; (obtained) 178.1 (MH) + .
步驟3：1－甲基－1H－苯幷咪唑－5－基胺(250) 依照流程25步驟2(實例64)所述之程序藉由催化氫化硝基化合物249獲得化合物250。LRMS：(計算值)147.2；(獲得值)148.1(MH)＋ 。 Step 3: 1-Methyl-1H-benzimidazole-5-ylamine (250) Compound 250 was obtained by catalytic hydrogenation of nitro compound 249 according to the procedure described in Scheme 25, Step 2 (Example 64). LRMS: (calculated) 147.2; (obtained) 148.1 (MH) + .
步驟4：4－[(1－甲基－1H－苯幷咪唑－5－基胺基)－甲基]－苯甲酸(251) 依照流程3步驟2(實例12)所述之程序，藉由使胺250與4－甲醯基－苯甲酸反應獲得標題化合物251。1 H NMR,(DMSO)δ(ppm)：8.24(s,1H),7.89(d,J＝7.9 Hz,2H),7.49(d,J＝7.9 Hz,2H),7.35(d,J＝8.9 Hz,1H),6.80(d,J＝8.9 Hz,1H),6.61(s,1H),4.39(s,2H),3.77(s,3H)。LRMS：(計算值)281.3；(獲得值)282.3(MH)＋ 。 Step 4: 4-[(1-Methyl-1H-benzimidazol-5-ylamino)-methyl]-benzoic acid (251) according to the procedure described in Step 3 (Example 12) of Scheme 3, Reaction of the amine 250 with 4-methylmercapto-benzoic acid afforded the title compound 251. 1 H NMR, (DMSO) δ (ppm): 8.24 (s, 1H), 7.89 (d, J = 7.9 Hz, 2H), 7.49 (d, J = 7.9 Hz, 2H), 7.35 (d, J = 8.9) Hz, 1H), 6.80 (d, J = 8.9 Hz, 1H), 6.61 (s, 1H), 4.39 (s, 2H), 3.77 (s, 3H). LRMS: (calculated) 281.3; (obtained) 282.3 (MH) + .
步驟5：N－(2－胺基－苯基)－4－[(1－甲基－1H－苯幷咪唑－5－基胺基)－甲基]－苯甲醯胺(255) 依照流程1步驟5(實例1)所述之程序藉由使酸251與1,2－苯二胺偶合獲得標題化合物252。1 H NMR：(DMSO)δ(ppm)：9.57(s,1H),8.00(s,1H),7.91(d,J＝8.4 Hz,2H),7.51(d,J＝8.4 Hz,2H),7.28(d,J＝8.4 Hz,1H),7.15(d,J＝7.5 Hz,1H),6.96(t,J＝7.5 Hz,1H),6.76(m,2H),6.62(s,1H),6.57(d,J＝7.5 Hz,1H),6.17(bs,1H),5.01(bs,2H),4.40(s,2H),3.73(s,3H)。LRMS：(計算值)371.4；(獲得值)372.4(MH)＋ 。 Step 5: N-(2-Amino-phenyl)-4-[(1-methyl-1H-benzoimidazole-5-ylamino)-methyl]-benzamide (255) according to the scheme The procedure described in 1 step 5 (Example 1) gave the title compound 252 by coupling the acid 251 with 1,2-phenylenediamine. 1 H NMR: (DMSO) δ (ppm): 9.57 (s, 1H), 8.00 (s, 1H), 7.91 (d, J = 8.4 Hz, 2H), 7.51 (d, J = 8.4 Hz, 2H), 7.28 (d, J = 8.4 Hz, 1H), 7.15 (d, J = 7.5 Hz, 1H), 6.96 (t, J = 7.5 Hz, 1H), 6.76 (m, 2H), 6.62 (s, 1H), 6.57 (d, J = 7.5 Hz, 1H), 6.17 (bs, 1H), 5.01 (bs, 2H), 4.40 (s, 2H), 3.73 (s, 3H). LRMS: (calculated) 371.4; (obtained) 372.4 (MH) + .
類似於實例134(化合物252)根據流程53以相應醯基氯代替原甲酸三甲酯製備實例134－140(化合物253－259)。Examples 134-140 (compounds 253-259) were prepared analogously to Example 134 (Compound 252) according to Scheme 53 substituting the corresponding decyl chloride for trimethyl orthoformate.
N－(2－胺基－苯基)－4－(苯幷噻唑－6－基胺甲基)－苯甲醯胺(263)N-(2-Amino-phenyl)-4-(benzothiazole-6-ylaminomethyl)-benzimidamide (263)
步驟1：苯幷噻唑－6－基胺(261) 依照流程33化合物143(實例79)所述之程序，藉由以氯化錫(II)還原硝基化合物260獲得標題化合物261。LRMS：(計算值)150.2；(獲得值)151.1(MH)＋ 。 Step 1: Benzothiazole-6-ylamine (261) The title compound 261 was obtained by the reduction of the nitro compound 260 with tin(II) chloride, according to the procedure of the compound LRMS: (calculated) 150.2; (obtained) 151.1 (MH) + .
步驟2：4－(苯幷噻唑－6－基胺甲基)－苯甲酸(262) 依照流程3步驟2(實例12)所述之程序，藉由使胺261 與4－甲醯基－苯甲酸反應獲得標題化合物262。1 H NMR,(DMSO)δ(ppm)：8.04(s,1H),7.90(d,J＝8.1 Hz,2H),7.74(d,J＝8.8 Hz,1H),7.49(d,J＝8.1 Hz,2H),7.07(d,J＝1.8 Hz,1H),6.89(dd,J＝1.8,8.1 Hz,1H),4.42(s,2H).LRMS：(計算值)284.3；(獲得值)285.2(MH)＋ 。 Step 2: 4-(benzothiazole-6-ylaminomethyl)-benzoic acid (262) according to the procedure described in Scheme 2, Step 2 (Example 12), by substituting amine 261 with 4-methylmercapto-benzene The formic acid reaction gave the title compound 262. 1 H NMR, (DMSO) δ (ppm): 8.04 (s, 1H), 7.90 (d, J = 8.1 Hz, 2H), 7.74 (d, J = 8.8 Hz, 1H), 7.49 (d, J = 8.1 Hz, 2H), 7.07 (d, J = 1.8 Hz, 1H), 6.89 (dd, J = 1.8, 8.1 Hz, 1H), 4.42 (s, 2H). LRMS: (calculated) 284.3; (obtained value) 285.2 (MH) + .
步驟3：－(2－胺基－苯基)－4－(苯幷噻唑－6－基胺甲基)－苯甲醯胺(263) 依照流程1步驟5(實例1)所述之程序，藉由使酸262與1,2－苯二胺偶合獲得標題化合物263。1 H NMR：(DMSO)δ(ppm)：9.59(s,1H),8.89(s,1H),7.93(d,J＝8.0 Hz,2H),7.75(d,J＝8.0 Hz,1H),7.50(d,J＝8.0 Hz,2H),7.14(d,J＝7.5 Hz,1H),7.08(s,1H),6.96(t,J＝7.5 Hz,1H),6.90(d,J＝8.0 Hz,1H),6.76(m,2H),6.58(s,1H),4.87(bs,2H),4.43(bs,2H).LRMS：(計算值)374.5；(獲得值)375.4(MH)＋ 。 Step 3: -(2-Amino-phenyl)-4-(benzothiazole-6-ylaminomethyl)-benzamide (263) according to the procedure described in Step 1 (Example 1) of Scheme 1, The title compound 263 was obtained by coupling the acid 262 with 1,2-phenylenediamine. 1 H NMR: (DMSO) δ (ppm): 9.59 (s, 1H), 8.89 (s, 1H), 7.93 (d, J = 8.0 Hz, 2H), 7.75 (d, J = 8.0 Hz, 1H), 7.50 (d, J = 8.0 Hz, 2H), 7.14 (d, J = 7.5 Hz, 1H), 7.08 (s, 1H), 6.96 (t, J = 7.5 Hz, 1H), 6.90 (d, J = 8.0) Hz, 1H), 6.76 (m, 2H), 6.58 (s, 1H), 4.87 (bs, 2H), 4.43 (bs, 2H). LRMS: (calculated) 374.5; (obtained value) 375.4 (MH) + .
N－(2－胺基－苯基)－4－[(1－甲基－1H－苯幷***－5－基胺基)－甲基]－苯甲醯胺(267)步驟1：1－甲基－5－硝基－1H－苯幷***(264) 在0℃下以NaNO2 (586 mg，8.5 mmol)之水(10 mL)溶液處理二胺248(1.13 g，6.76 mmol)及濃HCl(5.6 mL，67 mmol)經攪拌之水懸浮液。在相同條件下攪拌該混合物3小時，將其溫至室溫，以5% w/v KOH之水溶液中和且過濾之。以冷水洗滌及乾燥該固體產生標題化合物264(975 mg，產率81%)。1 H NMR：(DMSO)δ(ppm)：9.00(d,J＝1.3 Hz,1H),8.39(dd,J＝1.3,8.8 Hz,1H),8.09(d,J＝8.8 Hz,1H),4.40(s,3H)。 N-(2-Amino-phenyl)-4-[(1-methyl-1H-benzotriazol-5-ylamino)-methyl]-benzamide (267) Step 1:1 - methyl-5-nitro-benzene -1H- Bing-triazole (264) at 0 ℃ to NaNO 2 (586 mg, 8.5 mmol ) water (10 mL) was treated with a diamine 248 (1.13 g, 6.76 mmol) A stirred aqueous suspension of concentrated HCl (5.6 mL, 67 mmol). The mixture was stirred for 3 hours under the same conditions, warmed to room temperature, neutralized with an aqueous solution of 5% w/v KOH and filtered. The title compound 264 (975 mg, yield 81%). 1 H NMR: (DMSO) δ (ppm): 9.00 (d, J = 1.3 Hz, 1H), 8.39 (dd, J = 1.3, 8.8 Hz, 1H), 8.09 (d, J = 8.8 Hz, 1H), 4.40 (s, 3H).
步驟2：1－甲基－1H－苯幷***－5－基胺(265) 依照流程33化合物143(實例79)所述之程序藉由以氯化錫(II)還原硝基化合物264獲得標題化合物265。1 H NMR：(CD3 OD)δ(ppm)：7.32(d,J＝8.8 Hz,1H),7.14(d,J＝1.7 Hz,1H),6.99(dd,J＝1.7,8.8 Hz,1H),4.21(s,3H).LRMS：(計算值)148.3；(獲得值)149.3(MH)＋ 。 Step 2: 1-Methyl-1H-benzotriazol-5-ylamine (265) was obtained by reducing the nitro compound 264 with tin (II) chloride according to the procedure described for Compound 143 (Example 79). Title Compound 265. 1 H NMR: (CD 3 OD) δ (ppm): 7.32 (d, J = 8.8 Hz, 1H), 7.14 (d, J = 1.7 Hz, 1H), 6.99 (dd, J = 1.7, 8.8 Hz, 1H ), 4.21 (s, 3H). LRMS: (calculated) 148.3; (obtained value) 149.3 (MH) + .
步驟3：4－[(1－甲基－1H－苯幷***－5－基胺基)－甲基]－苯甲酸(266) 依照流程3步驟2(實例12)所述之程序，藉由使胺265與4－甲醯基－苯甲酸反應獲得標題化合物266。LRMS：(計算值)282.3；(獲得值)283.3(MH)＋ 。 Step 3: 4-[(1-Methyl-1H-benzotriazol-5-ylamino)-methyl]-benzoic acid (266) was obtained according to the procedure described in Step 3 (Example 12) of Scheme 3 The title compound 266 was obtained by reacting amine 265 with 4-methane-benzoic acid. LRMS: (calculated) 282.3; (obtained) 283.3 (MH) + .
步驟4：N－(2－胺基－苯基)－4－[(1－甲基－1H－苯幷***－5－基胺基)－甲基]－苯甲醯胺(267) 依照流程1步驟5(實例1)所述之程序，藉由使酸266與1,2－苯二胺偶合獲得標題化合物267。1 H NMR：(DMSO)δ(ppm)：9.58(s,1H),7.92(d,J＝8.0 Hz,2H),7.53(m,3H),7.14(d,J＝8.0 Hz,1H),7.07(dd,J＝2.0,8.5 Hz,1H),6.95(t,J＝7.5 Hz,1H),6.76(d,J＝8.0 Hz,1H),6.68(s,1H),6.61(s,2H),4.87(bs,2H),4.42(d,J＝6.0 Hz,2H),4.16(s,3H).LRMS：(計算值)372.4；(獲得值)373.5(MH)＋ 。 Step 4: N-(2-Amino-phenyl)-4-[(1-methyl-1H-benzotriazol-5-ylamino)-methyl]-benzamide (267) The procedure described in Scheme 1, Step 5 (Example 1), gave the title compound 267 by coupling the acid 266 with 1,2-phenylenediamine. 1 H NMR: (DMSO) δ (ppm): 9.58 (s, 1H), 7.92 (d, J = 8.0 Hz, 2H), 7.53 (m, 3H), 7.14 (d, J = 8.0 Hz, 1H), 7.07 (dd, J=2.0, 8.5 Hz, 1H), 6.95 (t, J = 7.5 Hz, 1H), 6.76 (d, J = 8.0 Hz, 1H), 6.68 (s, 1H), 6.61 (s, 2H) ), 4.87 (bs, 2H), 4.42 (d, J = 6.0 Hz, 2H), 4.16 (s, 3H). LRMS: (calculated) 372.4; (obtained value) 373.5 (MH) + .
N－(2－胺基－苯基)－4－[(1－甲基－2－甲胺基－1H－苯幷咪唑－5－基胺基)－甲基]－苯甲醯胺(271)步驟1：甲基－(1－甲基－5－硝基－1H－苯幷咪唑－2－基)－胺(268) 以異硫氰酸甲酯(970 mg，12.9 mmol)處理二胺248(1.88 g，11.2 mmol)之吡啶(20 mL)溶液且在80℃下攪拌該混合物30分鐘，將其冷卻至15℃，以固態EDC(3.03 g，15.8 mmol，1.40當量)處理且繼續在80℃下加熱16小時。真空下移除吡啶後，由急驟層析法(溶離劑為CH2 Cl2 中之5% MeOH)純化該殘餘物產生標題化合物268(1.44 g，產率62%)。1 H NMR：(CD3 OD)δ(ppm)：8.12(d,J＝2.2 Hz,1H),7.94(dd,J＝2.2,8.8 Hz,1H),7.04(d,J＝8.8 Hz,1H),4.44(bs,2H),3.51(s,3H),3.04(s,3H).LRMS：(計算值)206.2；(獲得值)207.1(MH)＋ 。 N-(2-Amino-phenyl)-4-[(1-methyl-2-methylamino-1H-benzoimidazole-5-ylamino)-methyl]-benzamide (271 Step 1: methyl-(1-methyl-5-nitro-1H-benzimidazol-2-yl)-amine (268 ) treated with diisothiocyanate (970 mg, 12.9 mmol) 248 (1.88 g, 11.2 mmol) in pyridine (20 mL) EtOAc EtOAc (EtOAc) (EtOAc (EtOAcjjjjjjjj Heat at 80 ° C for 16 hours. After removal of pyridine in vacuo, purified by flash chromatography (eluent of 5% MeOH CH 2 Cl 2 in the) The residue was purified to give the title compound 268 (1.44 g, yield 62%). 1 H NMR: (CD 3 OD) δ (ppm): 8.12 (d, J = 2.2 Hz, 1H), 7.94 (dd, J = 2.2, 8.8 Hz, 1H), 7.04 (d, J = 8.8 Hz, 1H) ), 4.44 (bs, 2H), 3.51 (s, 3H), 3.04 (s, 3H). LRMS: (calculated) 206.2; (obtained value) 207.1 (MH) + .
步驟2：甲基－(1－甲基－5－胺基－1H－苯幷咪唑－2－基)－胺(269) 依照流程25步驟2(實例64)所述之程序，藉由催化氫化硝基化合物268獲得標題化合物269。1 H NMR：(CDCl3 )δ(ppm)：6.70(s,1H),6.62(d,J＝7.9 Hz,1H),6.29(d,J＝7.9 Hz,1H),5.97(bs,1H),3.63(bs,2H),3.15(s,3H),2.90(s,3H).LRMS：(計算值)176.2；(獲得值)177.3(MH)＋ 。 Step 2: Methyl-(1-methyl-5-amino-1H-benzimidazol-2-yl)-amine (269) by catalytic hydrogenation according to the procedure described in Scheme 25, Step 2 (Example 64) The title compound 269 was obtained from the nitro compound 268. 1 H NMR: (CDCl 3 ) δ (ppm): 6.70 (s, 1H), 6.62 (d, J = 7.9 Hz, 1H), 6.29 (d, J = 7.9 Hz, 1H), 5.97 (bs, 1H) , 3.63 (bs, 2H), 3.15 (s, 3H), 2.90 (s, 3H). LRMS: (calculated) 176.2; (obtained) 177.3 (MH) + .
步驟3：4－[(1－甲基－2－甲胺基－1H－苯幷咪唑－5－基胺基)－甲基]－苯甲酸(270) 依照流程3步驟2(實例12)所述之程序，藉由使胺269與4－甲醯基－苯甲酸反應獲得標題化合物270。1 H NMR：(DMSO)δ(ppm)：8.03(bs,1H),7.88(d,J＝7.9 Hz,2H),7.47(d,J＝7.9 Hz,2H),7.02(d,J＝8.3 Hz,1H),6.46(d,J＝8.3 Hz,1H),6.44(s,1H),4.35(bs,2H),3.43(s,3H),2.90(d,J＝3.5 Hz,3H).LRMS：(計算值)310.3；(獲得值)311.4(MH)＋ 。 Step 3: 4-[(1-Methyl-2-methylamino-1H-benzimidazol-5-ylamino)-methyl]-benzoic acid (270) according to Scheme 3, Step 2 (Example 12) In the procedure described, the title compound 270 is obtained by reacting the amine 269 with 4-methylmercapto-benzoic acid. 1 H NMR: (DMSO) δ (ppm): 8.03 (bs, 1H), 7.88 (d, J = 7.9 Hz, 2H), 7.47 (d, J = 7.9 Hz, 2H), 7.02 (d, J = 8.3 Hz, 1H), 6.46 (d, J = 8.3 Hz, 1H), 6.44 (s, 1H), 4.35 (bs, 2H), 3.43 (s, 3H), 2.90 (d, J = 3.5 Hz, 3H). LRMS: (calculated) 310.3; (obtained) 311.4 (MH) + .
步驟4：N－(2－胺基－苯基)－4－[(1－甲基－2－甲胺基－1H－苯幷咪唑－5－基胺基)－甲基]－苯甲醯胺(271) 依照流程1步驟5(實例1)所述之程序藉由使酸270與1,2－苯二胺偶合獲得標題化合物271。1 H NMR：(DMSO)δ(ppm)：9.56(s,1H),7.90(d,J＝7.9 Hz,2H),7.49(d,J＝7.9 Hz,2H),7.15(d,J＝7.5 Hz,1H),6.80(d,J＝8.4 Hz,1H),6.76(d,J＝8.4 Hz,1H),6.58(t,J＝7.5 Hz,1H),6.39(s,1H),6.31(m,2H),5.75(t,J＝5.7 Hz,1H),4.87(s,2H),4.32(d,J＝5.7 Hz,2H),3.34(s,3H),2.82(d,J＝4.4 Hz,3H).LRMS：(計算值)400.5；(獲得值)401.5(MH)＋ 。 Step 4: N-(2-Amino-phenyl)-4-[(1-methyl-2-methylamino-1H-benzoimidazole-5-ylamino)-methyl]-benzamide The title compound 271 was obtained by coupling the acid 270 with 1,2-phenylenediamine according to the procedure described in Step 1 (Example 1). 1 H NMR: (DMSO) δ (ppm): 9.56 (s, 1H), 7.90 (d, J = 7.9 Hz, 2H), 7.49 (d, J = 7.9 Hz, 2H), 7.15 (d, J = 7.5) Hz, 1H), 6.80 (d, J = 8.4 Hz, 1H), 6.76 (d, J = 8.4 Hz, 1H), 6.58 (t, J = 7.5 Hz, 1H), 6.39 (s, 1H), 6.31 ( m, 2H), 5.75 (t, J = 5.7 Hz, 1H), 4.87 (s, 2H), 4.32 (d, J = 5.7 Hz, 2H), 3.34 (s, 3H), 2.82 (d, J = 4.4 Hz, 3H). LRMS: (calculated) 400.5; (obtained value) 401.5 (MH) + .
N－(2－胺基－苯基)－4－(喹噁啉－6－基胺甲基)－苯甲醯胺(276)步驟1：6－硝基－噻噁啉(273) 以40%乙二醛水溶液(0.85 mL，7.4 mmol，1.1當量)(或任意其它之1,2－二羰基化合物，1.1當量)處理硝基苯胺272(1.04 g，6.76 mmol)之2－丙醇(35 mL)溶液。在80℃下攪拌該混合物2小時且將其在真空下濃縮產生標題化合物273，其未經進一步純化即用於下一步驟中。LRMS：(計算值)175.1；(獲得值)176.1(MH)＋ 。 N-(2-Amino-phenyl)-4-(quinoxalin-6-ylaminemethyl)-benzamide (276) Step 1: 6-Nitro- thiamethane (273) at 40 Treatment of nitroaniline 272 (1.04 g, 6.76 mmol) of 2-propanol (35 g) with aqueous glyoxal (0.85 mL, 7.4 mmol, 1.1 eq.) (or any other 1,2-dicarbonyl compound, 1.1 eq.) mL) solution. The mixture was stirred at <RTI ID=0.0></RTI></RTI><RTIgt; LRMS: (calculated) 175.1; (obtained) 176.1 (MH) + .
步驟2：喹噁啉－6－基胺(274) 依照與流程33化合物143(實例79)所述相同之程序藉由以氯化錫(II)還原硝基化合物273獲得標題化合物274。LRMS：(計算值)145.2；(獲得值)146.2(MH)＋ 。 Step 2: Quinoxaline-6-ylamine (274) The title compound 274 was obtained by the reduction of the nitro compound 273 with tin(II) chloride. LRMS: (calculated) 145.2; (obtained) 146.2 (MH) + .
步驟3：4－(噻噁啉－6－基胺甲基)－苯甲酸(275) 依照流程3步驟2(實例12)所述之程序，藉由使胺274與4－甲醯基－苯甲酸反應獲得標題化合物275。LRMS：(計算值)279.3；(獲得值)280.2(MH)＋ 。 Step 3: 4-(Thio-oxalin-6-ylaminomethyl)-benzoic acid (275) according to the procedure described in Step 2, Step 2 (Example 12), by subjecting amine 274 to 4-carbamido-benzene The formic acid reaction gave the title compound 275. LRMS: (calculated) 279.3; (obtained) 280.2 (MH) + .
步驟4：N－(2－胺基－苯基)－4－(喹噁啉－6－基胺甲基)－苯甲醯胺(276) 依照流程1步驟5(實例1)所述之程序藉由使酸275與1,2－苯二胺偶合獲得標題化合物276。1 H NMR：(DMSO)δ(ppm)：9.61(s,1H),8.56(d,J＝2.0 Hz,1H),8.43(d,J＝2.0 Hz,1H),7.93(d,J＝8.0 Hz,2H),7,75(d,J＝9.0 Hz,1H),7.52(d,J＝8.0 Hz,2H),7.40－7.36(m,2H),7.13(dd,J＝1.6,6.8 Hz,1H),6.95(dt,J＝1.6,8.0 Hz,1H),6.76(dd,J＝1.2,7.8 Hz,1H),6.67(d,J＝2.5 Hz,1H),6.59(dd,J＝1.2,7.8 Hz,1H),5.05(bs,2H),4.53(d,J＝5.7 Hz,2H).LRMS：(計算值)369.4；(獲得值)370.4(MH)＋ 。 Step 4: N-(2-Amino-phenyl)-4-(quinoxalin-6-ylaminomethyl)-benzamide (276) according to the procedure described in Step 1 (Example 1) of Scheme 1. The title compound 276 was obtained by coupling the acid 275 with 1,2-phenylenediamine. 1 H NMR: (DMSO) δ (ppm): 9.61 (s, 1H), 8.56 (d, J = 2.0 Hz, 1H), 8.43 (d, J = 2.0 Hz, 1H), 7.93 (d, J = 8.0 Hz, 2H), 7, 75 (d, J = 9.0 Hz, 1H), 7.52 (d, J = 8.0 Hz, 2H), 7.40-7.36 (m, 2H), 7.13 (dd, J = 1.6, 6.8 Hz , 1H), 6.95 (dt, J = 1.6, 8.0 Hz, 1H), 6.76 (dd, J = 1.2, 7.8 Hz, 1H), 6.67 (d, J = 2.5 Hz, 1H), 6.59 (dd, J = 1.2, 7.8 Hz, 1H), 5.05 (bs, 2H), 4.53 (d, J = 5.7 Hz, 2H). LRMS: (calculated) 369.4; (obtained value) 370.4 (MH) + .
N－(2－胺基－苯基)－4－[(2,3－二－吡啶－2－基－喹噁啉－6－基胺基)－甲基]－苯甲醯胺(277) 依照流程57實例144所述之程序，第一步中使用1,2－二吡啶－2－基－乙烷－1,2－二酮代替乙二醛製備標題化合物。 N-(2-Amino-phenyl)-4-[(2,3-di-pyridin-2-yl-quinoxalin-6-ylamino)-methyl]-benzamide (277) The title compound was prepared in the first step using 1,2-dipyridin-2-yl-ethane-1,2-dione in place of glyoxal according to the procedure described in Example 144 of Scheme 57.
4－((2－(2－(二甲胺基)乙基)－1,3－二酮基異吲哚啉－6－基胺基)甲基)－N－(2－胺基苯基)苯甲醯胺(278)步驟1. 2－(2－(二甲胺基)乙基)－5－硝基異吲哚啉－1,3－二酮(280) 以純N1,N1－二甲基乙烷－1,2－二胺(0.75 mL；5.8 mmol；1.13 當量)(或相應胺，1.3當量)處理硝基鄰苯二甲酸酐(279 )(995 mg；5.2 mmol)之乙酸(12 mL)溶液。在100℃下攪拌該反應混合物3小時，將其冷卻至室溫，在真空下濃縮；使殘餘物溶解於乙酸乙酯(250 mL)中且以飽和NaHCO3 進行洗滌，經MgSO4 乾燥，過濾且濃縮之產生呈黃色固體之化合物280 (1.19 g；4.5 mmol；87%)。LRMS：263.3(計算值)；264.2(獲得值)(MH)＋ 。 4-((2-(2-(Dimethylamino)ethyl)-1,3-diketoisoindoline-6-ylamino)methyl)-N-(2-aminophenyl) Benzoguanamine (278) Step 1. 2-(2-(Dimethylamino)ethyl)-5-nitroisoindoline-1,3-dione (280) as pure N1, N1-di Treatment of nitrophthalic anhydride ( 279 ) (995 mg; 5.2 mmol) of acetic acid with methyl ethane-1,2-diamine (0.75 mL; 5.8 mmol; 1.13 eq.) (or corresponding amine, 1.3 eq.) 12 mL) solution. The mixture was stirred for 3 hours at 100 deg.] C the reaction, it was cooled to room temperature, concentrated in vacuo; the residue was dissolved in ethyl acetate (250 mL) and the washed with saturated NaHCO 3, dried over MgSO 4, filtered Concentration gave Compound 280 (1.19 g; 4.5 mmol; 87%). LRMS: 263.3 (calculated); 264.2 (obtained) (MH) + .
步驟2. 5－胺基－2－(2－(二甲胺基)乙基)異吲哚啉－1,3－二酮(281) 依照流程25步驟2(實例64)所述之程序，藉由催化氫化硝基化合物280獲得標題化合物281。LRMS：233.3(計算值)；234.2(獲得值)(MH)＋ 。 Step 2. 5-Amino-2-(2-(dimethylamino)ethyl)isoindoline-1,3-dione (281) according to the procedure described in Scheme 25, Step 2 (Example 64), The title compound 281 was obtained by catalytic hydrogenation of the nitro compound 280. LRMS: 233.3 (calculated); 234.2 (obtained) (MH) + .
步驟3. 4－((2－(2－(二甲胺基)乙基)－1,3－二酮基異吲哚啉－6－基胺基)甲基)苯甲酸(282) 依照流程3步驟2(實例12)所述之程序，藉由使胺281與4－甲醯基－苯甲酸反應獲得標題化合物282。1 H NMR,(DMSO)δ(ppm)：7.89(d,J＝8.2 Hz,2H),7.78(t,J＝6.1 Hz,1H),7.53(d,J＝8.2 Hz,1H),7.43(d,J＝8.2 Hz,2H),6.91(s,1H),4.53(d,J＝6.1 Hz,2H),3.80(t,J＝5.5 Hz,2H),3.32(bs,2H),3.21(bs,2H),2.74(s,6H).LRMS：367.4(計算值)；368.4(獲得值)(MH)＋ 。 Step 3. 4-((2-(2-(Dimethylamino)ethyl)-1,3-dioneisoindoline-6-ylamino)methyl)benzoic acid (282) according to the scheme The procedure described in 3 Step 2 (Example 12) gave the title compound 282 by reacting amine 281 with 4-methane-benzoic acid. 1 H NMR, (DMSO) δ (ppm): 7.89 (d, J = 8.2 Hz, 2H), 7.78 (t, J = 6.1 Hz, 1H), 7.53 (d, J = 8.2 Hz, 1H), 7.43 ( d, J = 8.2 Hz, 2H), 6.91 (s, 1H), 4.53 (d, J = 6.1 Hz, 2H), 3.80 (t, J = 5.5 Hz, 2H), 3.32 (bs, 2H), 3.21 ( Bs, 2H), 2.74 (s, 6H). LRMS: 367.4 (calculated); 368.4 (obtained) (MH) + .
步驟4：4－((2－(2－(二甲胺基)乙基)－1,3－二酮基異吲哚啉－6－基胺基)甲基)－N－(2－胺基苯基)苯甲醯胺(278) 依照流程1步驟5(實例1)所述之程序藉由使酸282與1,2－苯二胺偶合獲得標題化合物278。1 H NMR：(DMSO)δ(ppm)：9.52(s,1H),7.85(d,J＝8.0 Hz,2H),7.63(t,J＝5.9 Hz,1H),7.42(d,J＝8.2 Hz,2H),7.37(d,J＝8.2 Hz,2H),7.05(d,J＝7.2 Hz,1H),6.87(d,J＝7.2 Hz,1H),6.82(s,1H),6.76(d,J＝8.2 Hz,2H),6.67(d,J＝7.8 Hz,1H),6.49(t,J＝7.2 Hz,1H),4.81(s,2H),4.45(d,J＝5.9 Hz,2H),3.48(t,J＝6.3 Hz,2H),2.32(t,J＝6.3 Hz,2H),2.04(s,6H).LRMS：(計算值)457.5；(獲得值)458.5(MH)＋ 。 Step 4: 4-((2-(2-(Dimethylamino)ethyl)-1,3-dioneisoindoline-6-ylamino)methyl)-N-(2-amine The phenylphenyl)benzamide (278) was obtained by coupling the acid 282 with 1,2-phenylenediamine according to the procedure described in Step 1 (Example 1). 1 H NMR: (DMSO) δ (ppm): 9.52 (s, 1H), 7.85 (d, J = 8.0 Hz, 2H), 7.63 (t, J = 5.9 Hz, 1H), 7.42 (d, J = 8.2 Hz, 2H), 7.37 (d, J = 8.2 Hz, 2H), 7.05 (d, J = 7.2 Hz, 1H), 6.87 (d, J = 7.2 Hz, 1H), 6.82 (s, 1H), 6.76 ( d, J = 8.2 Hz, 2H), 6.67 (d, J = 7.8 Hz, 1H), 6.49 (t, J = 7.2 Hz, 1H), 4.81 (s, 2H), 4.45 (d, J = 5.9 Hz, 2H), 3.48 (t, J = 6.3 Hz, 2H), 2.32 (t, J = 6.3 Hz, 2H), 2.04 (s, 6H). LRMS: (calculated) 457.5; (obtained value) 458.5 (MH) + .
N－(2－胺基－苯基)－4－[(1,3－二酮基－2－吡啶－3－基甲基－2,3－二氫－1H－異吲哚－5－基胺基)－甲基]－苯甲醯胺(283) 根據流程58對實例146所述之反應順序，但在第一步中以3－胺甲基吡啶代替N,N－二甲基乙二胺製備標題化合物。 N-(2-Amino-phenyl)-4-[(1,3-diketo-2-pyridin-3-ylmethyl-2,3-dihydro-1H-isoindole-5-yl) Amino)-methyl]-benzamide (283) according to Scheme 58 for the reaction sequence described in Example 146, but in the first step, 3-aminomethylpyridine was substituted for N,N-dimethylethylene. The title compound was prepared as an amine.
N－(2－胺基－苯基)－4－{[4－(2,4－二甲基－噻唑－5－基)－嘧啶－2－基胺基]－甲基}－苯甲醯胺(314)N-(2-Amino-phenyl)-4-{[4-(2,4-dimethyl-thiazol-5-yl)-pyrimidin-2-ylamino]-methyl}-benzamide Amines (314)
根據流程6類似於化合物26a(實例29)用1－(2,4－二甲基－噻唑－5－基)－乙酮(表11)代替1－吡嗪－2－基－乙酮作為原料獲得標題化合物。標題化合物之表徵結果提供於表12中。Substituting 1-(2,4-dimethyl-thiazol-5-yl)-ethanone (Table 11) for 1-pyrazin-2-yl-ethanone according to Scheme 6 analogous to compound 26a (Example 29) The title compound was obtained. The characterization results of the title compounds are provided in Table 12.
N－(2－胺基－苯基)－4－{[4－(2H－吡唑－3－基)－嘧啶－2－基胺基]－甲基}－苯甲醯胺(315)N-(2-Amino-phenyl)-4-{[4-(2H-pyrazol-3-yl)-pyrimidin-2-ylamino]-methyl}-benzamide (315)
根據流程6類似於化合物26a(實例29)用1－(2H－吡唑－3－基)－乙酮(表11)代替1－吡嗪－2－基－乙酮作為原料獲得標題化合物。標題化合物之表徵結果提供於表12中。The title compound was obtained by substituting 1-(2H-pyrazol-3-yl)-ethanone (Table 11) for 1-pyridazin-2-yl-ethanone as a starting material. The characterization results of the title compounds are provided in Table 12.
N－(2－胺基－苯基)－4－{[4－[2,4－二甲基－噁唑－5－基]－嘧啶－2－基胺基]－甲基}－苯甲醯胺(316)N-(2-Amino-phenyl)-4-{[4-[2,4-dimethyl-oxazol-5-yl]-pyrimidin-2-ylamino]-methyl}-benzene Guanamine (316)
根據流程6類似於化合物26a(實例29)用1－(2,4－二甲基－噁唑－5－基)－乙酮(表11)代替1－吡嗪－2－基－乙酮作為原料獲得標題化合物。標題化合物之表徵結果提供於表12中。Substituting 1-(2,4-dimethyl-oxazol-5-yl)-ethanone (Table 11) for 1-pyrazin-2-yl-ethanone according to Scheme 6 analogous to compound 26a (Example 29) The title compound was obtained as a starting material. The characterization results of the title compounds are provided in Table 12.
N－(2－胺基－苯基)－4－{[4－(2,4－二甲基－噁唑－5－基)－嘧啶－2－基胺基]－甲基}－苯甲醯胺(317)N-(2-Amino-phenyl)-4-{[4-(2,4-dimethyl-oxazol-5-yl)-pyrimidin-2-ylamino]-methyl}-benzene Guanamine (317)
步驟1：5－(2－甲基－1,3－二氧雜環戊烷－2－基)異噁唑－3－甲酸乙酯(318) 使由苯(50 mL)中之5－乙醯基異噁唑－3－甲酸乙酯(2.53 g，13.8 mmol)、乙二醇(1.29 g，20.7 mmol)及p－TsOH(0.13 g，0.69 mmol)組成之反應混合物在丁史塔克(Dean－Stark)接受管中回流24小時(流程59)。減壓移除大部分溶劑且使殘餘物在飽和NaHCO3 與EtOAc之間分溶。收集有機層，將其以鹽水、水洗滌且經MgSO4 乾燥。減壓蒸發經乾燥之萃取物產生呈油狀物之標題化合物(3.14 g，產率100%)，該化合物未經進一步純化即用於下一步驟中。1 H NMR(DMSO－d6 )δ(ppm)：6.89(s,1H),4.35(q,J＝7.2 Hz,2H),4.07－4.01(m,2H),4.0l－3.94(m,2H),1.70(s,3H),1.32(t,J＝7.0 Hz,3H).MS(m/z)：227.21(計算值)228.1(MH＋ )(實驗值)。 Step 1: 5-(2-Methyl-1,3-dioxolan-2-yl)isoxazole-3-carboxylic acid ethyl ester (318) from 5-B in benzene (50 mL) A reaction mixture of mercaptoisoxazole-3-carboxylic acid ethyl ester (2.53 g, 13.8 mmol), ethylene glycol (1.29 g, 20.7 mmol) and p-TsOH (0.13 g, 0.69 mmol) in Ding Stark ( Dean-Stark) was subjected to reflux in the tube for 24 hours (Scheme 59). Most of the solvent is removed and the residue was partitioned between saturated NaHCO 3 and EtOAc under reduced pressure. The organic layer was collected, which with brine, washed with water and dried over MgSO 4. The dried <RTI ID=0.0></RTI><RTIID=0.0> 1 H NMR (DMSO-d 6 ) δ (ppm): 6.89 (s, 1H), 4.35 (q, J = 7.2 Hz, 2H), 4.07-4.01 (m, 2H), 4.0l - 3.94 (m, 2H) ), 1.70 (s, 3H) , 1.32 (t, J = 7.0 Hz, 3H) .MS (m / z): 227.21 ( calc) 228.1 (MH +) (Found).
步驟2：(5－(2－甲基－1,3－二氧雜環戊烷－2－基)異噁唑－3－基)甲醇(319) 向溶於EtOH－THF為1：2之混合物(45 mL)中的二氧雜環戊烷318(3.14 g，13.8 mmol)溶液中加入NaBH4 (0.68 g，18.0 mmol)。在室溫下攪拌該反應混合物2小時，以水處理之且蒸發有機溶劑。以EtOAc萃取含水相且相繼以H2 O及鹽水洗滌經合併之有機層，使其經MgSO4 乾燥且減壓濃縮產生呈油狀物之標題化合物(2.39 g，產率93%)，該化合物未經進一步純化即用於下一步驟中。1 H NMR(DMSO－d6 )δ(ppm)：6.43(s,1H),5.47(t,J＝5.9 Hz,1H),4.47(d,J＝5.9 Hz,2H),4.06－4.02(m,2H),3.94－3.90(m,2H),1.66(s,3H).MS(m/z)：185.18(計算值)186.1(MH＋ )(實驗值)。 Step 2: (5-(2-Methyl-1,3-dioxolan-2-yl)isoxazol-3-yl)methanol (319) is dissolved in EtOH-THF at 1:2 dioxolane mixture (45 mL) of 318 (3.14 g, 13.8 mmol) was added NaBH 4 (0.68 g, 18.0 mmol ). The reaction mixture was stirred at room temperature for 2 hours, treated with water and evaporated organic solvent. The aqueous phase was extracted with EtOAc and sequentially washed with H 2 O and brine and the organic layers were combined, dried and concentrated under reduced pressure over MgSO 4 produced the title compound as an oil (2.39 g, 93% yield) of the compound Used in the next step without further purification. 1 H NMR (DMSO-d 6 ) δ (ppm): 6.43 (s, 1H), 5.47 (t, J = 5.9 Hz, 1H), 4.47 (d, J = 5.9 Hz, 2H), 4.06 - 4.02 (m) , 2H), 3.94-3.90 (m, 2H), 1.66 (s, 3H) .MS (m / z): 185.18 ( calc) 186.1 (MH +) (Found).
步驟3. 1－(3－(羥甲基)異噁唑－5－基)乙酮(320) 向原醇319(2.39 g，12.9 mmol)之MeOH(30 mL)溶液中加入10% HCl(30 mL)。在70℃下攪拌該反應混合物18小時，將其冷卻且以1 M NaOH溶液中和至pH 6。蒸發MeOH且以EtOAc萃取所得含水相。以鹽水洗滌有機層，將其在MgSO4 下乾燥且減壓濃縮產生呈米色固體之標題化合物(1.67 g，產率92%)，該化合物未經進一步純化即用於下一步驟中。1 H NMR(DMSO－d6 )δ(ppm)：7.25(s,1H),5.62(t,J＝6.1 Hz,1H),4.56(d,J＝5.9 Hz,2H),2.56(s,3H).MS(m/z)：141.12(計算值)142.1(MH＋ )(實驗值)。 Step 3. 1-(3-(Hydroxymethyl)isoxazol-5-yl)ethanone (320) To a solution of hexanes (30 mL) (3. mL). The reaction mixture was stirred at 70 ° C for 18 hours, cooled and neutralized to pH 6 with 1 M NaOH solution. The MeOH was evaporated and the resulting aqueous phase was extracted with EtOAc. The organic layer was washed with brine, which was dried under MgSO 4 and concentrated under reduced pressure to produce the title compound of white solid (1.67 g, yield 92%) which was used without further purification in the next step. 1 H NMR (DMSO-d 6 ) δ (ppm): 7.25 (s, 1H), 5.62 (t, J = 6.1 Hz, 1H), 4.56 (d, J = 5.9 Hz, 2H), 2.56 (s, 3H) MS (m/z): 141.12 (calculated) 142.1 (MH + ) (exp.).
步驟4：N－(2－胺基－苯基)－4－{[4－(2,4－二甲基－噁唑－5－基)－嘧啶－2－基胺基]－甲基}－苯甲醯胺(317) 根據流程6類似於化合物26a(實例29)用酮320(表11)代替1－吡嗪－2－基－乙酮作為原料獲得標題化合物。標題化合物之結構及表徵結果示於表12中。 Step 4: N-(2-Amino-phenyl)-4-{[4-(2,4-dimethyl-oxazol-5-yl)-pyrimidin-2-ylamino]-methyl} - Benzylamine (317) The title compound was obtained according to the procedure 6 to Compound 26a (Example 29) using ketone 320 (Table 11) instead of 1-pyrazin-2-yl-ethanone as starting material. The structure and characterization results of the title compound are shown in Table 12.
N－(2－胺基苯基)－4－((4－(3－(羥甲基)－5－甲基異噁唑－4－基)嘧啶－2－基胺基)甲基)苯甲醯胺(321)N-(2-Aminophenyl)-4-((4-(3-(hydroxymethyl)-5-methylisoxazol-4-yl)pyrimidin-2-ylamino)methyl)benzene Formamide (321)
步驟1：5－甲基－4－(2－甲基－1,3－二氧雜環戊烷－2－基)－異噁唑－3－甲酸甲酯(322) 根據流程60，類似於二氧雜環戊烷318以82%之產率獲得標題化合物。MS(m/z)：227.21(計算值)228.1(MH＋ )(實驗值)。 Step 1: methyl 5-methyl-4-(2-methyl-1,3-dioxolan-2-yl)-isoxazole-3-carboxylate (322) according to Scheme 60, similar The dioxolane 318 gave the title compound in 82% yield. MS (m / z): 227.21 ( calc) 228.1 (MH +) (Found).
步驟2：(5－甲基－4－(2－甲基－1,3－二氧雜環戊烷－2－基)異噁唑－3－基)甲醇(323) 根據流程60，類似於原醇319以94%之產率獲得標題化合物。1 H NMR(DMSO－d6 )δ(ppm)：5.21(t,J＝5.7 Hz,1H),4.48(d,J＝5.7 Hz,2H),3.98－3.94(m,2H),3.71－3.67(m,2H),2.39(s,3H),1.60(s,3H).MS(m/z)：199.20(計算值)200.1(MH＋ )(實驗值)。 Step 2: (5-Methyl-4-(2-methyl-1,3-dioxolan-2-yl)isoxazol-3-yl)methanol (323) according to Scheme 60, analogous The original alcohol 319 gave the title compound in a yield of 94%. 1 H NMR (DMSO-d 6 ) δ (ppm): 5.21 (t, J = 5.7 Hz, 1H), 4.48 (d, J = 5.7 Hz, 2H), 3.98-3.94 (m, 2H), 3.71-3.67 (m, 2H), 2.39 ( s, 3H), 1.60 (s, 3H) .MS (m / z): 199.20 ( calc) 200.1 (MH +) (Found).
步驟3：1－(3－(羥甲基)－5－甲基異噁唑－4－基)乙酮(324) 根據流程60類似於酮320以77%之產率獲得標題化合物。1 H NMR(DMSO－d6 )δ(ppm)：5.45(t,J＝5.9 Hz,1H),4.66(d,J＝5.9 Hz,2H),2.66(s,3H),2.51(s,3H).MS(m/z)：155.15(計算值)156.1(MH＋ )(實驗值)。 Step 3: 1-(3-(Hydroxymethyl)-5-methylisoxazol-4-yl)ethanone (324) mp . 1 H NMR (DMSO-d 6 ) δ (ppm): 5.45 (t, J = 5.9 Hz, 1H), 4.66 (d, J = 5.9 Hz, 2H), 2.66 (s, 3H), 2.51 (s, 3H) MS (m/z): 155.15 (calculated) 156.1 (MH + ) (found).
步驟4：N－(2－胺基苯基)－4－((4－(3－(羥甲基)－5－甲基異噁唑－4－基)嘧啶－2－基胺基)甲基)苯甲醯胺(317) 根據流程6類似於化合物26a(實例29)用酮324(表11)代替1－吡嗪－2－基－乙酮作為原料獲得標題化合物。標題化合物之結構及表徵結果提供於表12中。 Step 4: N-(2-Aminophenyl)-4-((4-(3-(hydroxymethyl)-5-methylisoxazol-4-yl)pyrimidin-2-ylamino) A Benzobenzamide (317) The title compound was obtained according to the procedure of Compound 6 (Comp. 29) using ketone 324 (Table 11) instead of 1-pyrazin-2-yl-ethanone. The structure and characterization results of the title compounds are provided in Table 12.
N－(2－胺基苯基)－4－((4－(1－甲基－1H－1,2,3－***－4－基)嘧啶－2－基胺基)甲基)苯甲醯胺(325)及N－(2－胺基苯基)－4－((4－(3－甲基－3H－1,2,3－***－4－基)嘧啶－2－基胺基)甲基)苯甲醯胺(326)N-(2-Aminophenyl)-4-((4-(1-methyl-1H-1,2,3-triazol-4-yl)pyrimidin-2-ylamino)methyl)benzene Formamide (325) and N-(2-aminophenyl)-4-((4-(3-methyl-3H-1,2,3-triazol-4-yl)pyrimidin-2-yl) Amino)methyl)benzamide (326)
步驟1：1－(3H－1,2,3－***－4－基)乙酮(327) 向3－丁炔－2－酮(627 mg，9.21 mmol)之二甲苯(10 mL)溶液中加入疊氮三丁基錫(4.00 g，12.0 mmol)。在一密閉燒瓶中在140℃下攪拌該反應混合物3小時。蒸發二甲苯且由急驟層析法以EtOAc為溶離劑純化該殘餘物產生標題化合物(645 mg，產率63%)。1 H NMR(DMSO－d6 )δ(ppm)：8.51(s,1H),2.56(s,3H).MS(m/z)：111.10(計算值)112.1(MH＋ )(實驗值)。 Step 1: 1-(3H-1,2,3-triazol-4-yl)ethanone (327) to 3-butyn-2-one (627 mg, 9.21 mmol) in xylene (10 mL) Tributyltin azide (4.00 g, 12.0 mmol) was added. The reaction mixture was stirred at 140 ° C for 3 hours in a closed flask. The residue was purified by flash chromatography eluting EtOAc elut elut elut 1 H NMR (DMSO-d 6 ) δ (ppm): 8.51 (s, 1H), 2.56 (s, 3H) .MS (m / z): 111.10 ( calc) 112.1 (MH +) (Found).
步驟2：(E)－3－(二甲胺基)－1－(1－甲基－1H－1,2,3－***－4－基)丙－2－烯－1－酮(328)及(E)－3－(二甲胺基)－1－(3－甲基－3H－1,2,3－***－4－基)丙－2－烯－1－酮(329) 根據流程6類似於化合物23a(實例29，步驟1)使用酮327(表11)代替1－吡嗪－2－基－乙酮作為原料獲得標題化合物328與329之混合物(產率24%)。MS(m/z)：180.21(計算值)181.1(MH＋ )(實驗值)。 Step 2: (E)-3-(Dimethylamino)-1-(1-methyl-1H-1,2,3-triazol-4-yl)prop-2-en-1-one (328 And (E)-3-(dimethylamino)-1-(3-methyl-3H-1,2,3-triazol-4-yl)prop-2-en-1-one (329) A mixture of the title compound 328 and 329 (yield 24%) was obtained from the compound </ br></RTI></RTI></RTI></RTI><RTIgt; MS (m / z): 180.21 ( calc) 181.1 (MH +) (Found).
1 H NMR(DMSO－d6 )δ(ppm)(328，主要異構體，試驗測定)：8.01(s,1H),7.74(d,J＝12.3 Hz,1H),5.73(d,J＝12.3 Hz,1H),4.18(s,3H),3.15(s,3H),2.88(s,3H)。 1 H NMR (DMSO-d 6 ) δ (ppm) (328, major isomer, determined by test): 8.01 (s, 1H), 7.74 (d, J = 12.3 Hz, 1H), 5.73 (d, J = 12.3 Hz, 1H), 4.18 (s, 3H), 3.15 (s, 3H), 2.88 (s, 3H).
1 H NMR(DMSO－d6 )δ(ppm)(329，次要異構體，試驗測定)：8.26(s,1H),7.71(d,J＝10.8 Hz,1H),5.66(d,J＝12.1 Hz,1H),4.19(s,3H),3.16(s,3H),2.92(s,3H)。 1 H NMR (DMSO-d 6 ) δ (ppm) (329, minor isomer, assay): 8.26 (s, 1H), 7.71 (d, J = 10.8 Hz, 1H), 5.66 (d, J) = 12.1 Hz, 1H), 4.19 (s, 3H), 3.16 (s, 3H), 2.92 (s, 3H).
步驟3：4－((4－(1－甲基－1H－1,2,3－***－4－基)嘧啶－2－基胺基)甲基)苯甲酸(330)及4－((4－(3－甲基－3H－1,2,3－***－4－基)嘧啶－2－基胺基)甲基)苯甲酸(331) 根據流程6類似於化合物25a(實例29，步驟3)使用烯胺酮328及329之混合物代替烯胺酮23a作為原料以80%之產率獲得標題化合物之混合物。1 H NMR(DMSO－d6 )δ(ppm)(328，主要異構體，試驗性分配)：12.80(s,1H),8.33(d,J＝5.1 Hz,1H),8.17(s,1H),7.94(t,J＝6.5 Hz,1H),7.85(d,J＝8.2 Hz,2H),7.42(d,J＝8.2 Hz,2H),7.02(d,J＝4.7 Hz,1H),4.60(d,J＝6.5 Hz,2H),4.22(s,3H).MS(m/z)：310.31(計算值)311.2(MH＋ )(實驗值)。 Step 3: 4-((4-(1-methyl-1H-1,2,3-triazol-4-yl)pyrimidin-2-ylamino)methyl)benzoic acid (330) and 4-( (4-(3-Methyl-3H-1,2,3-triazol-4-yl)pyrimidin-2-ylamino)methyl)benzoic acid (331) is similar to compound 25a according to Scheme 6 (Example 29) Step 3) Using a mixture of enaminone 328 and 329 in place of enaminone 23a as a starting material, a mixture of the title compounds was obtained in a yield of 80%. 1 H NMR (DMSO-d 6 ) δ (ppm) (328, major isomer, experimental partition): 12.80 (s, 1H), 8.33 (d, J = 5.1 Hz, 1H), 8.17 (s, 1H) ), 7.94 (t, J = 6.5 Hz, 1H), 7.85 (d, J = 8.2 Hz, 2H), 7.42 (d, J = 8.2 Hz, 2H), 7.02 (d, J = 4.7 Hz, 1H), 4.60 (d, J = 6.5 Hz, 2H), 4.22 (s, 3H). MS (m/z): 310.31 (calc.) 311.2 (MH + ) (the experimental value).
步驟4：N－(2－胺基苯基)－4－((4－(1－甲基－1H－1,2,3－***－4－基)嘧啶－2－基胺基)甲基)苯甲醯胺(325)及N－(2－胺基苯基)－4－((4－(3－甲基－3H－1,2,3－***－4－基)嘧啶－2－基胺基)甲基)苯甲醯胺(326) 根據流程6類似於化合物26a(實例29，步驟4)使用酸330及331之混合物代替酸25a作為原料以53%及7%之產率獲得標題化合物。標題化合物之表徵結果提供於表12中。 Step 4: N-(2-Aminophenyl)-4-((4-(1-methyl-1H-1,2,3-triazol-4-yl)pyrimidin-2-ylamino) A Benzomethane (325) and N-(2-aminophenyl)-4-((4-(3-methyl-3H-1,2,3-triazol-4-yl)pyrimidine- 2-Aminoamino)methyl)benzamide (326) is similar to compound 26a (Example 29, Step 4) using a mixture of acids 330 and 331 in place of acid 25a as a starting material at 53% and 7% according to Scheme 6. Rate to obtain the title compound. The characterization results of the title compounds are provided in Table 12.
N－(2－胺基苯基)－4－((4－(2－甲基咪唑幷[1,2－a]吡啶－3－基)嘧啶－2－基胺基)甲基)苯甲醯胺(332)N-(2-Aminophenyl)-4-((4-(2-methylimidazolium[1,2-a]pyridin-3-yl)pyrimidin-2-ylamino)methyl)benzene Guanamine (332)
步驟1：1－(2－甲基咪唑幷[1,2－a]吡啶－3－基)乙酮(333) 向戊－2,4－二酮(1.00 g,9.99 mmol)及N－溴代丁二醯亞胺(1.96 g，10.99 mmol)之CHCl3 (20 mL)溶液中加入1,1'－偶氮雙(環己腈)(催化量)。攪拌該反應混合物1小時，使其經過濾且減壓濃縮濾液。將殘餘物再溶解於THF/Et2 O為1：1之混合物(20 mL)中，接著添加吡啶－2－胺(723 mg，7.68 mmol)且使反應混合物回流隔夜。冷卻後減壓移除溶劑且以EtOAc接著EtOAc－MeOH(96：4)為溶離劑由柱層析法純化該殘餘物產生標題化合物(475 mg，產率35%)。1 H NMR(DMSO－d6 )δ(ppm)：9.59(dt,J＝6.8,1.4 Hz,1H),7.68(dt,J＝8.8,1.2 Hz,1H),7.56(ddd,J＝8.8,6.8,1.4 Hz,1H),7.17(td,J＝6.8,1.4 Hz,1H),2.72(s,3H),2.58(s,3H).MS(m/z)：174.20(計算值)175.1(MH＋ )(實驗值)。 Step 1: 1-(2-methylimidazolium [1,2-a]pyridin-3-yl)ethanone (333) to penta-2,4-dione (1.00 g, 9.99 mmol) and N-bromo substituting succinic acyl imine (1.96 g, 10.99 mmol) of CHCl 3 (20 mL) was added 1,1'-azobis (cyclohexanecarbonitrile) (catalytic amount). The reaction mixture was stirred for 1 h, filtered and the filtrate evaporated. The residue was redissolved in THF / Et 2 O 1: a mixture of 1 (20 mL) followed by addition of pyridin-2-amine (723 mg, 7.68 mmol) and the reaction mixture was refluxed overnight. After cooling, the solvent was evaporated,jjjjjjjjjjjjj 1 H NMR (DMSO-d 6 ) δ (ppm): 9.59 (dt, J = 6.8, 1.4 Hz, 1H), 7.68 (dt, J = 8.8, 1.2 Hz, 1H), 7.56 (ddd, J = 8.8, 6.8, 1.4 Hz, 1H), 7.17 (td, J = 6.8, 1.4 Hz, 1H), 2.72 (s, 3H), 2.58 (s, 3H). MS (m/z): 174.20 (calculated) 175.1 ( MH + ) (experimental value).
[M.Anderson,J.F.Beattie等人，Bioorg.Med.Chem.Lett.；2003,13；3021－3026]。[M. Anderson, J. F. Beattie et al, Bioorg. Med. Chem. Lett.; 2003, 13; 3021-3026].
步驟2：N－(2－胺基苯基)－4－((4－(2－甲基咪唑幷[1,2－a]吡啶－3－基)嘧啶－2－基胺基)甲基)苯甲醯胺(332) 根據流程6類似於化合物26a(實例29)使用酮333(表11)代替1－吡嗪－2－基－乙酮作為原料獲得標題化合物。標題化合物之表徵結果提供於表12中。 Step 2: N-(2-Aminophenyl)-4-((4-(2-methylimidazolium[1,2-a]pyridin-3-yl)pyrimidin-2-ylamino)methyl Benzoguanamine (332) was obtained according to Scheme 6 analogous to Compound 26a (Example 29) using ketone 333 (Table 11) instead of 1-pyrazin-2-yl-ethanone as the starting material. The characterization results of the title compounds are provided in Table 12.
4－((4－(2－胺基－4－甲基噻唑－5－基)嘧啶－2－基胺基)甲基)－N－(2－胺基苯基)苯甲醯胺(334)及N－(2－胺基苯基)－4－((4－(5－(2－(二甲胺基)乙醯胺基)－3－甲基噻吩－2－基)嘧啶－2－基胺基)甲基)苯甲醯胺(335)4-((4-(2-Amino-4-methylthiazol-5-yl)pyrimidin-2-ylamino)methyl)-N-(2-aminophenyl)benzamide (334 And N-(2-aminophenyl)-4-((4-(5-(2-(dimethylamino)ethyl)amino)-3-methylthiophen-2-yl)pyrimidine-2 -ylamino)methyl)benzamide (335)
步驟1：2－(雙－第三丁氧基羰基－胺基－)－5－乙醯基－4－甲基噻唑(336) 向Boc2 O(3.07 g，14.1 mmol)及1－(2－胺基－4－甲基噻唑－5－基)(2.00 g，12.8 mmol)之DCM(20 mL)溶液中加入吡啶(1.11 g，14.1 mmol)。在室溫下攪拌該反應混合物3天。加入相同量之Boc2 O且再攪拌反應混合物3天。減壓蒸發DCM，添加水且以EtOAc萃取所得混合物。以鹽水洗滌有機層，使其經MgSO4 乾燥且減壓濃縮以產生呈橙色油狀物之標題化合物(4.6 g，產率100%)。1 H NMR(DMSO－d6 )δ(ppm)：2.54(s,3H),2.49(s,3H),1.53(s,18H).MS(m/z)：356.44(計算值)357.1(MH＋ )(實驗值)。 Step 1: 2-(Bis-tert-butoxycarbonyl-amino-)-5-ethenyl-4-methylthiazole (336) to Boc 2 O (3.07 g, 14.1 mmol) and 1-(2) Pyridine (1.11 g, 14.1 mmol) was added to a solution of <RTI ID=0.0></RTI></RTI><RTIgt;</RTI><RTIgt; The reaction mixture was stirred at room temperature for 3 days. The same amount of Boc 2 O was added and the reaction mixture was stirred for another 3 days. The DCM was evaporated under reduced pressure, water was added andEtOAc. The organic layer was washed with brine, it was dried over MgSO 4, and concentrated under reduced pressure to yield an orange oil of the title compound (4.6 g, 100% yield). 1 H NMR (DMSO-d 6 ) δ (ppm): 2.54 (s, 3H), 2.49 (s, 3H), 1.53 (s, 18H). MS (m/z): 356.44 (calc.) 357.1 (MH) + ) (experimental value).
步驟2：(E)－1－[2－(雙－第三丁氧基羰基－胺基－)－4－甲基噻唑－5－基]－3－(二甲胺基)丙－2－烯－1－酮(337) 依照用於合成烯胺酮23a所述之程序(流程6)但以酮336代替1－(吡嗪－2－基)－乙酮以16%之產率獲得標題化合物。1 H NMR(DMSO－d6 )δ(ppm)：7.65(d,J＝12.1 Hz,1H),5.34(d,J＝12.1 Hz,1H),3.14(s,3H),2.87(s,3H),2.50(s,3H),1.51(s,18H).Ms(m/z)：411.52(計算值)412.3(MH＋ )(實驗值)。 Step 2: (E)-1-[2-(bis-tert-butoxycarbonyl-amino-)-4-methylthiazol-5-yl]-3-(dimethylamino)propan-2- The en-1-one (337) was obtained according to the procedure described for the synthesis of enaminone 23a (Scheme 6) but with ketone 336 instead of 1-(pyrazin-2-yl)-ethanone in 16% yield. Compound. 1 H NMR (DMSO-d 6 ) δ (ppm): 7.65 (d, J = 12.1 Hz, 1H), 5.34 (d, J = 12.1 Hz, 1H), 3.14 (s, 3H), 2.87 (s, 3H) ), 2.50 (s, 3H) , 1.51 (s, 18H) .Ms (m / z): 411.52 ( calc) 412.3 (MH +) (Found).
步驟3：(E)－1－[2－(第三丁氧基羰基－胺基－)－4－甲基噻唑－5－基]－3－(二甲胺基)丙－2－烯－1－酮(338) 以NaOMe溶液(25% ww，1.9 ml)處理烯胺酮337(859 mg，2.09 mmol)之甲醇(12 mL)溶液。使反應混合物回流24小時，以NaOH溶液(1 M，3 ml)進行處理，使其冷卻至室溫，以1 M HCl小心中和(pH 7.5－8)且以EtOAc萃取之。經MgSO4 乾燥萃取物，使其經過濾及蒸發提供相應於標題化合物之殘餘物(721 mg，大於定量產量)，該殘餘物未經進一步純化即用於下一步驟中。MS(m/z)：311.40(計算值)312.1(MH＋ )(實驗值)。 Step 3: (E)-1-[2-(Tertibutoxycarbonyl-amino-)-4-methylthiazol-5-yl]-3-(dimethylamino)prop-2-ene- 1-ketone (338) A solution of enaminone 337 (859 mg, 2.09 mmol) in methanol (12 mL) was taken from NaOMe (25% ww, 1.9 ml). The reaction mixture was refluxed for 24 h, then EtOAc EtOAc (EtOAc) The extract was dried over MgSO 4, it was filtered and evaporated to provide the title compound of the residue (721 mg, greater than quantitative yield), the residue was used without further purification in the next step. MS (m / z): 311.40 ( calc) 312.1 (MH +) (Found).
步驟4：4－((4－(2－(第三丁氧羰基胺基)－4－甲基噻唑－5－基)嘧啶－2－基胺基)甲基)苯甲酸(339) 依照用於合成酸25a所述之程序(流程6，步驟3)但以烯胺酮338代替烯胺酮23a以18%之產率獲得標題化合物，且該標題化合物未經進一步純化即用於後續步驟中。MS(m/z)：441.50(計算值)442.3(MH＋ )(實驗值)。 Step 4: 4-((4-(2-(Tertidinoxycarbonylamino)-4-methylthiazol-5-yl)pyrimidin-2-ylamino)methyl)benzoic acid (339) The title compound was obtained in the same procedure as the procedure for the synthesis of the acid 25a (Scheme 6, Step 3), but the enaminone 338 was replaced by the enaminone 23a in 18% yield, and the title compound was used in the next step without further purification. . MS (m / z): 441.50 ( calc) 442.3 (MH +) (Found).
步驟5及6：4－((4－(2－胺基－4－甲基噻唑－5－基)嘧啶－2－基胺基)甲基)－N－(2－胺基苯基)苯甲醯胺(334) 根據用於合成化合物26a所述之程序(流程6，步驟4，與1,2－苯二胺偶合)隨後以對合成化合物117(流程28，步驟5，胺基去保護反應)所述之程序獲得標題化合物。兩步之產率為78%。標題化合物之表徵結果提供於表12中。 Steps 5 and 6: 4-((4-(2-Amino-4-methylthiazol-5-yl)pyrimidin-2-ylamino)methyl)-N-(2-aminophenyl)benzene Formamide (334) is then deprotected according to the procedure described for the synthesis of compound 26a (Scheme 6, Step 4, coupled with 1,2-phenylenediamine) followed by the synthesis of Compound 117 (Scheme 28, Step 5, Amino Group) Reaction of the procedure described to obtain the title compound. The yield in two steps was 78%. The characterization results of the title compounds are provided in Table 12.
步驟7：4－((4－(2－胺基－4－甲基噻唑－5－基)嘧啶－2－基胺基)甲基)苯甲酸(340) 根據對合成化合物117所述之程序(流程28，步驟5，胺基去保護反應)獲得定量產量之標題化合物(純度約90%)。MS(m/z)：341.39(計算值)342.1(MH＋ )(實驗值)。 Step 7: 4-((4-(2-Amino-4-methylthiazol-5-yl)pyrimidin-2-ylamino)methyl)benzoic acid (340) according to the procedure described for the synthesis of compound 117 (Scheme 28, Step 5, Amino Deprotection) The title compound was obtained in quantitative yield (purity: about 90%). MS (m / z): 341.39 ( calc) 342.1 (MH +) (Found).
步驟8：4－((4－(2－((二甲胺基)甲基胺甲醯基)－4－甲基噻唑－5－基)嘧啶－2－基胺基)甲基)苯甲酸(341) 將二甲胺基乙醯氯氫氯酸鹽(59.0 mg，0.37 mmol)加入至酸340(98.1 mg，0.29 mmol)之吡啶(5 mL)溶液中。在室溫下攪拌反應混合物一天接著添加另一份二甲胺基乙醯氯氫氯酸鹽(40 mg，0.12 mmol)且在40℃下再攪拌該混合物一天。減壓蒸發吡啶且添加MeOH。形成固體材料，將其經過濾收集且經製備型RP HPLC(AQUASIL C－18柱；5μM；230 x 21.2 mm；溶離劑水中之20－80% MeOH)純化以產生24.5 mg標題化合物(產率20%)。MS(m/z)：426.49(計算值)427.2(MH＋ )(實驗值)。 Step 8: 4-((4-(2-Dimethylamino)methylaminemethanyl)-4-methylthiazol-5-yl)pyrimidin-2-ylamino)methyl)benzoic acid (341) Dimethylaminoethyl chlorohydrin hydrochloride (59.0 mg, 0.37 mmol) was added to a solution of EtOAc (EtOAc (EtOAc) The reaction mixture was stirred at room temperature for one day and then another portion of dimethylaminoacetic acid hydrochloride (40 mg, 0.12 mmol) was added and the mixture was stirred at 40 ° C for one day. The pyridine was evaporated under reduced pressure and MeOH was added. A solid material was formed which was collected by filtration and purified by preparative RP HPLC (AQUASIL C-18 column; 5 [mu]M; 230 x 21.2 mm; 20-80% MeOH in solvent water) to yield 24.5 mg of the title compound (yield 20 %). MS (m / z): 426.49 ( calc) 427.2 (MH +) (Found).
步驟9：N－(2－胺基苯基)－4－((4－(5－(2－(二甲胺基)乙醯胺基)－3－甲基噻吩－2－基)嘧啶－2－基胺基)甲基)苯甲醯胺(335) 根據用於合成化合物26a所述之程序(流程6，步驟4)獲得產率為15%之標題化合物(經製備型HPLC純化，AQUASIL C－18柱；5μM；230 x 21.2 mm；溶離劑20－80%水中之MeOH)。標題化合物之表徵結果提供於表12中。 Step 9: N-(2-Aminophenyl)-4-((4-(5-(2-(dimethylamino)ethyl)amino)-3-methylthiophen-2-yl)pyrimidine- 2-Aminoamino)methyl)benzamide (335) gave the title compound as a 15% yield (purified by preparative HPLC, AQUASIL ) according to the procedure used for the synthesis of compound 26a (Scheme 6, Step 4) C-18 column; 5 μM; 230 x 21.2 mm; solvate 20-80% MeOH in water). The characterization results of the title compounds are provided in Table 12.
4－(4－(吡啶－3－基)嘧啶－2－基胺基)－N－(2－胺基苯基)苯甲醯胺(342)步驟1：4－(4－(吡啶－3－基)嘧啶－2－基胺基)苯甲酸(343) 根據用於合成化合物25a所述之程序(流程6，步驟3)，藉由以4－胍基苯甲酸(344)(Zlatoidsky P.,Maliar T.Eur,J.Med.Chem Chim.Ther.；1996,31,895－900)代替胍24且以(E)－3－(二甲胺基)－1－(吡啶－3－基)丙－2－烯－1－酮(345)(Zimmermann J.,Buchdunger E.等人，Bioorg.Med.Chem.Lett.,1996, 6,1221－1226)代替(E)－3－(二甲胺基)－1－(吡嗪－2－基)－丙－2－烯－1－酮(23a)製備標題化合物。產物之產率為28%。MS(m/z)：292.29(計算值)293.1(MH＋ )(實驗值)。 4-(4-(pyridin-3-yl)pyrimidin-2-ylamino)-N-(2-aminophenyl)benzamide (342) Step 1: 4-(4-(pyridine-3) -yl)pyrimidin-2-ylamino)benzoic acid (343) according to the procedure described for the synthesis of compound 25a (Scheme 6, step 3) by 4-mercaptobenzoic acid (344) (Zlatoidsky P. ,Maliar T.Eur,J.Med.Chem Chim.Ther.;1996,31,895-900) instead of 胍24 and (E)-3-(dimethylamino)-1-(pyridin-3-yl)propene 2-en-1-one (345) (Zimmermann J., Buchdunger E. et al., Bioorg. Med. Chem. Lett., 1996, 6 , 1221-1226) instead of (E)-3-(dimethylamine) The title compound was prepared as 1-(pyrazin-2-yl)-prop-2-en-1-one (23a). The yield of the product was 28%. MS (m / z): 292.29 ( calc) 293.1 (MH +) (Found).
步驟2：4－(4－(吡啶－3－基)嘧啶－2－基胺基)－N－(2－胺基苯基)苯甲醯胺(342) 根據用於合成化合物26a所述之程序(流程6，步驟4)以酸343代替酸25a獲得產率為54%之標題化合物。標題化合物之表徵結果提供於表12中。 Step 2: 4-(4-(pyridin-3-yl)pyrimidin-2-ylamino)-N-(2-aminophenyl)benzamide (342) according to the compound used for the synthesis of compound 26a The procedure (Scheme 6, Step 4) replaced the acid 25a with acid 343 to give the title compound in 54% yield. The characterization results of the title compounds are provided in Table 12.
N－(2－胺基－苯基)－4－(2,4－二酮基－亞噻唑烷－5－基甲基)－苯甲醯胺(346)步驟1：4－(2,4－二酮基－亞噻唑烷－5－基甲基)－苯甲酸(347) 使4－甲醯基－苯甲酸(1.0 g，6.7 mmol)、噻唑烷－2,4－二酮(0.78 g，6.7 mmol)及哌啶(1.32 ml，13.3 mmol)之2－甲氧基－乙醇(20 ml)溶液回流2小時，冷卻至室溫，將其蒸發且將含油殘餘物再溶解於水中。以濃HCl(pH 1－2)酸化該溶液產生沉澱，過濾收集該沉澱，將其乾燥且以熱丙酮濕磨以產生標題化合物(1.06 g，產率64%)。LRMS：249.2(計算值)，248.1[M－H]－ (實驗值)。 N-(2-Amino-phenyl)-4-(2,4-dione-thiathiazolidine-5-ylmethyl)-benzamide (346) Step 1: 4-(2,4 -Diketo-thiathiazolidine-5-ylmethyl)-benzoic acid (347) 4-methylmercapto-benzoic acid (1.0 g, 6.7 mmol), thiazolidine-2,4-dione (0.78 g) A solution of 6.7 mmol) and piperidine (1.32 ml, 13.3 mmol) in 2-methoxy-ethanol (20 ml) was refluxed for 2 hr, cooled to room temperature, evaporated and the oil residue was redissolved in water. The solution was acidified with cone. HCl (pH 1-2) to give crystals, which crystals crystals crystals crystals LRMS: 249.2 (calculated), 248.1 [M-H] - (exp.).
步驟2：{2－[4－(2,4－二酮基－亞噻唑烷－5－基甲基)－苯甲醯胺基]－苯基)－胺基甲酸第三丁酯(348) 依照用於合成化合物115所述之程序(流程28)但以酸347代替酸114獲得產率為63%之標題化合物。LRMS：439.5(計算值)，462.4[M＋Na]＋ (實驗值)。 Step 2: {2-[4-(2,4-Diketyl-thiazolidine-5-ylmethyl)-benzylideneamino]-phenyl)-carbamic acid tert-butyl ester (348) The title compound was obtained in a yield of 63% according to the procedure used for the synthesis of compound 115 (Scheme 28). LRMS: 439.5 (calculated), 462.4 [M+Na] + (found).
步驟3：N－(2－胺基－苯基)－4－(2,4－二酮基－亞噻唑烷－5－基甲基)－苯甲醯胺(346) 依照用於合成化合物117所述之程序(流程28)但以醯胺348代替醯胺116獲得產率為37%之標題化合物。1 H NMR：(400 MHz,DMSO－d6 ,δ(ppm)：9.72(s,1H),8.06(d,j＝8.2 Hz,2H),7.79(s,1H),7.69(d,J＝8.4,2H),7.14(d(dd)J＝7.8,1H),6.95(d(dd),J＝1.6 Hz,J＝9.0,1H),6.75(dd,J＝1.2 Hz,J＝8.0 Hz,1H),6.56(t(dd),J＝7.2 Hz 1H).LRMS：339.4(計算值)，340.4[MH]＋ (實驗值)。 Step 3: N-(2-Amino-phenyl)-4-(2,4-dione-thiathiazolidine-5-ylmethyl)-benzamide (346) was used to synthesize compound 117 The procedure described (Scheme 28) but substituting guanamine 348 for guanamine 116 afforded the title compound in 37% yield. 1 H NMR: (400 MHz, DMSO-d 6 , δ (ppm): 9.72 (s, 1H), 8.06 (d, j = 8.2 Hz, 2H), 7.79 (s, 1H), 7.69 (d, J = 8.4, 2H), 7.14 (d(dd)J=7.8, 1H), 6.95 (d(dd), J=1.6 Hz, J=9.0, 1H), 6.75 (dd, J=1.2 Hz, J=8.0 Hz , 1H), 6.56 (t (dd), J = 7.2 Hz 1H). LRMS: 339.4 (calculated), 340.4 [MH] + (exp.).
N－(2－胺基－苯基)－4－(2,4－二酮基－噻唑烷－5－基甲基)－苯甲醯胺(349)步驟1：4－(2,4－二酮基－亞噻唑烷－5－基甲基)－苯甲酸甲酯(350) 使4－甲醯基－苯甲酸甲酯(1.0 g，6.7 mmol)、噻唑烷－2,4－二酮(0.78 g，6.7 mmol)及哌啶(0.66 ml,6.7 mmol)之2－二甲氧基－乙醇(20 ml)溶液回流2小時，冷卻至室溫且將其蒸發。以CH2 Cl2 濕磨殘餘物產生結晶材料，過濾收集該材料產生標題化合物(620 mg，產率35%)。LRM：263.3(計算值)，264.1[MH]＋ (實驗值)。 N-(2-Amino-phenyl)-4-(2,4-dione-thiazolidine-5-ylmethyl)-benzamide (349) Step 1: 4-(2,4- Diketo-thiazolidine-5-ylmethyl)-benzoic acid methyl ester (350) 4-methylmercapto-benzoic acid methyl ester (1.0 g, 6.7 mmol), thiazolidine-2,4-dione (0.78 g, 6.7 mmol) and a solution of piperidine (0.66 ml, 6.7 mmol) in 2-dimethoxy-ethanol (20 ml) were refluxed for 2 hr, cooled to room temperature and evaporated. In CH 2 Cl 2 and the residue was triturated produce crystalline material, the material was collected by filtration to give the title compound (620 mg, 35% yield). LRM: 263.3 (calculated), 264.1 [MH] + (exp.).
步驟2：4－(2,4－二酮基－噻唑烷－5－基甲基)－苯甲酸甲酯(351) 室溫下使甲酯350(615 mg，2.32 mmol)之MeOH(120 ml)溶液經10% Pd/C(615 mg，德固賽(Degussa)型)氫化2小時。加入另一份Pd/C(300 mg)且繼續再進行氫化3小時(由MS監控)。經由矽藻土墊過濾反應混合物，將其蒸發且以EtOAc－CH2 Cl2 (1：2)為溶離劑由急驟層析法純化該殘餘物以產生標題化合物(570 mg，產率92%)。LRMS：265.3(計算值)，266.1[MH]＋ (實驗值)。 Step 2: 4-(2,4-dione-thiazolidin-5-ylmethyl)-benzoic acid methyl ester (351) mp 350 (615 mg, 2.32 mmol) MeOH (120 ml) The solution was hydrogenated with 10% Pd/C (615 mg, Degussa type) for 2 hours. Another portion of Pd/C (300 mg) was added and hydrogenation was continued for a further 3 hours (monitored by MS). The reaction mixture was filtered through a pad of diatomaceous earth, and evaporated to which EtOAc-CH 2 Cl 2 (1 : 2) as eluent The residue was purified by flash chromatography to give the title compound (570 mg, 92% yield) . LRMS: 265.3 (calculated), 266.1 [MH] + (found).
步驟3：4－(2,4－二酮基－噻唑烷－5－基甲基)－苯甲酸(352) 以濃HCl(5 ml)處理甲酯351(250 mg，0.94 mmol)之AcOH(10 ml)溶液且將反應混合物在120℃下加熱2小時，加以冷卻及蒸發產生固體殘餘物，將該殘餘物再懸浮於水中且由過濾收集產生標題化合物(98 mg，產率41%)。LRMS：251.3(計算值)，250.1[M－H]－ (實驗值)。 Step 3: 4- (2,4-diketo - thiazolidin-5-ylmethyl) - benzoic acid (352) with concentrated HCl (5 ml) methyl process 351 (250 mg, 0.94 mmol) of AcOH ( The title compound (98 mg, yield 41%) was obtained from EtOAc. LRMS: 251.3 (calculated), 250.1 [M-H] - (found).
步驟4：N－(2－胺基－苯基)－4－(2,4－二酮基－噻唑烷－5－基甲基)－苯甲醯胺(349) 依照用於合成化合物10a所述之程序(流程2，實例2)但以酸352代替酸9 獲得產率為51%之標題化合物。1 H NMR：(400 MHz,DMSO－d6 ,δ(ppm)：12.04(br s,1H),9.62(s,1H),7.90(d,j＝8.2 Hz,2H),7.36(d,J＝8.2,2H),7.16(d(dd)J＝8.4,1H),7.13(d(dd),J＝6.7,1H),6.95(dd,J＝1.6 Hz,J＝7.8 Hz,1H),6.75(dd,j＝1.4 Hz,j＝8.0 Hz,1H),6.57(dd,j＝1.4 Hz,7.6 Hz,1H),4.99(dd,J＝4.7 Hz,J＝9.0 Hz,1H),4.96(br s,2H),3.45(dd,J＝4.5 Hz,J＝14.1 Hz,1H),3.22(dd,J＝9.0 Hz,J＝14.3 Hz,1H).LRMS：341.3(計算值),342.3[MH]＋ (實驗值)。 Step 4: N-(2-Amino-phenyl)-4-(2,4-dione-thiazolidine-5-ylmethyl)-benzamide (349) according to the compound used for the synthesis of compound 10a The procedure described (Scheme 2, Example 2) but substituting acid 352 for acid 9 gave the title compound as a 51% yield. 1 H NMR: (400 MHz, DMSO-d 6 , δ (ppm): 12.04 (br s, 1H), 9.62 (s, 1H), 7.90 (d,j = 8.2 Hz, 2H), 7.36 (d, J) = 8.2, 2H), 7.16 (d (dd) J = 8.4, 1H), 7.13 (d (dd), J = 6.7, 1H), 6.95 (dd, J = 1.6 Hz, J = 7.8 Hz, 1H), 6.75 (dd, j = 1.4 Hz, j = 8.0 Hz, 1H), 6.57 (dd, j = 1.4 Hz, 7.6 Hz, 1H), 4.99 (dd, J = 4.7 Hz, J = 9.0 Hz, 1H), 4.96 (br s, 2H), 3.45 (dd, J = 4.5 Hz, J = 14.1 Hz, 1H), 3.22 (dd, J = 9.0 Hz, J = 14.3 Hz, 1H). LRMS: 341.3 (calculated), 342.3 [MH] + (experimental value).
(E)－N－(2－胺基苯基)－4－(2－(4－((2,4－二酮基亞噻唑烷－5－基)甲基)苯氧基)乙氧基)苯甲醯胺(353)步驟1：4－(2－溴乙氧基)苯甲醛(354) 在0℃下向K2 CO3 (4.52 g，32.7 mmol)、1,2－二溴乙烷(10.6 ml，122.8 mmol)之DMF(12 mL)懸浮液中加入4－羥基苯甲醛(1.0 g，8.2 mmol)之DMF(3 mL)溶液。在室溫下攪拌該混合物18小時，將其過濾且蒸發之。以EtOAc－己烷梯度(EtOAc百分比自20至25%增加)由矽膠柱層析法純化該殘餘物產生標題化合物(1.21 g，產率64%)。1 H NMR：(CDCl3 )δ(ppm)：9.88(s,1H),7.84(d,J＝8.0 Hz,2H),7.01(d,J＝8.0 Hz,2H),4.38(t,J＝6 Hz,2H),3.67(t,J＝6 Hz,2H).LRMS(ESI)：(計算值)227.9、229.9；(實驗值)229.1、231.3(MH)＋ 。 (E)-N-(2-Aminophenyl)-4-(2-(4-((2,4-dione)thiazolidine-5-yl)methyl)phenoxy)ethoxy Benzoylamine (353) Step 1: 4-(2-Bromoethoxy)benzaldehyde (354) to K 2 CO 3 (4.52 g, 32.7 mmol), 1,2-dibromoethane at 0 ° C A solution of 4-hydroxybenzaldehyde (1.0 g, 8.2 mmol) in DMF (3 mL). The mixture was stirred at room temperature for 18 hours, filtered and evaporated. The residue was purified by EtOAc EtOAc elut elut elut elut elut elut 1 H NMR: (CDCl 3 ) δ (ppm): 9.88 (s, 1H), 7.84 (d, J = 8.0 Hz, 2H), 7.01 (d, J = 8.0 Hz, 2H), 4.38 (t, J = 6 Hz, 2H), 3.67 (t, J = 6 Hz, 2H). LRMS (ESI): (calculated) 227.9, 229.9; (experimental) 229.1, 231.3 (MH) + .
步驟2：4－(2－(4－甲醯基苯氧基)乙氧基)苯甲酸甲酯(355) 向354(1.21 g，5.27 mmol)之DMF(10 mL)溶液中加入4－羥基苯甲酸甲酯(0.80 g,5.27 mmol)及K2 CO3 (2.91 g，21.1 mmol)。在60℃下攪拌所得混合物6小時，將其過濾及蒸發。以EtOAc－己烷(1：2)為溶離劑由矽膠柱層析法純化殘餘物產生標題化合物(0.83 g，53%)。LRMS(ESI)：(計算值)300.3；(實驗值)301.4(MH)＋ 。 Step 2: Add methyl 4-(2-(4-carbamidophenoxy)ethoxy)benzoate (355) to a solution of 354 (1.21 g, 5.27 mmol) in DMF (10 mL) Methyl benzoate (0.80 g, 5.27 mmol) and K 2 CO 3 (2.91 g, 21.1 mmol). The resulting mixture was stirred at 60 ° C for 6 hours, filtered and evaporated. The residue was purified by EtOAc EtOAc elut elut elut elut elut LRMS (ESI) :( calc) 300.3; (Found) 301.4 (MH) +.
步驟3：(E)－甲基－4－(2－(4－((2,4－二酮基亞噻唑烷－5－基)甲基)苯氧基)乙氧基)苯甲酸酯(356) 向355(1.59 g，3.86 mmol)之甲苯(10 mL)溶液中加入噻唑烷－2,4－二酮(542 mg，4.63 mmol)、苯甲酸(61.3 mg，0.50 mmol)及哌啶(57μL，0.58 mmol)。使所得混合物在Dean－Stark接受管中回流1/2小時且冷卻至室溫。形成沉澱物，其經過濾收集產生標題化合物(1.41 g，92%)。1 H NMR：(DMSO－d6 )δ(ppm)：7.90(d,J＝8.8 Hz,2H),7.73(s,1H),7.54(d,J＝8.8 Hz,2H),7.14(d,J＝8.8 Hz,2H),7.08(d J＝8.8 Hz,2H),4.42(bs,4H),3.80(s,3H).LRMS(ESI)：(計算值)399.1；(實驗值)400.0(MH)＋ 。 Step 3: (E)-Methyl-4-(2-(4-((2,4-dione)thiazolidin-5-yl)methyl)phenoxy)ethoxy)benzoate (356) Add thiazolidine-2,4-dione (542 mg, 4.63 mmol), benzoic acid (61.3 mg, 0.50 mmol) and piperidine to 355 (1.59 g, 3.86 mmol) in toluene (10 mL). (57 μL, 0.58 mmol). The resulting mixture was refluxed in a Dean-Stark receiver tube for 1/2 hour and cooled to room temperature. A precipitate formed which was collected by filtration afforded the title compound (l. 1 H NMR: (DMSO-d 6 ) δ (ppm): 7.90 (d, J = 8.8 Hz, 2H), 7.73 (s, 1H), 7.54 (d, J = 8.8 Hz, 2H), 7.14 (d, J = 8.8 Hz, 2H), 7.08 (d J = 8.8 Hz, 2H), 4.42 (bs, 4H), 3.80 (s, 3H). LRMS (ESI): (calculated) 399.1; (experimental value) 400.0 ( MH) + .
步驟4：(E)－4－(2－(4－((2,4－二酮基亞噻唑烷－5－基)甲基)苯氧基)乙氧基)苯甲酸(化合物357) 向甲酯356(647 mg，1.62 mmol)之THF(15 mL)溶液中加入甲醇(2 mL)、水(2 mL)及單水合氫氧化鋰(340 mg，8.11 mmol)。在60℃下加熱該混合物1小時，以10% HCl溶液酸化且以乙酸乙酯萃取之。使有機萃取物經MgSO4 乾燥，過濾及蒸發形成標題化合物(141 mg，22%)。1 H NMR：(MeOD－d4 )δ(ppm)：7.97(d,J＝8.8 Hz,2H),7.75(s,1H),7.53(d,J＝8.8 Hz,2H),7.12(d,J＝8.8 Hz,2H),7.04(d,J＝8.8 Hz,2H),4.44(4H,bs).LRMS(ESI)：(計算值)385.4；(實驗值)392.3(MLi)＋ 。 Step 4: (E)-4-(2-(4-((2,4-dione)thiazolidin-5-yl)methyl)phenoxy)ethoxy)benzoic acid (Compound 357) Methyl ester 356 (647 mg, 1.62 mmol) in THF (15 mL) MeOH (2 mL), EtOAc (EtOAc) The mixture was heated at 60 ° C for 1 hour, acidified with 10% HCl solution and extracted with ethyl acetate. The organic extracts were dried over MgSO 4, filtered and evaporated to provide the title compound (141 mg, 22%). 1 H NMR: (MeOD-d 4 ) δ (ppm): 7.97 (d, J = 8.8 Hz, 2H), 7.75 (s, 1H), 7.53 (d, J = 8.8 Hz, 2H), 7.12 (d, J = 8.8 Hz, 2H), 7.04 (d, J = 8.8 Hz, 2H), 4.44 (4H, bs). LRMS (ESI): (calc.) 385.4; (calc.) 392.3 (MLi) + .
步驟5：(E)－N－(2－胺基苯基)－4－(2－(4－(2,4－二酮基亞噻唑烷－5－基)甲基)苯氧基)乙氧基)苯甲醯胺(353) 將酸357(141 mg，0.37 mmol)、苯－1,2－二胺(40 mg，0.37 mmol)及BOP(161 mg，0.37 mmol)溶解於CH3 CN(5 mL)中。添加三乙胺(0.73 mmol，203μL)且在室溫下攪拌該反應18小時。減壓移除溶劑且以EtOAc－己烷混合物梯度溶離(EtOAc百分比自33%增加至100%)由矽膠柱層析法純化該殘餘物產生標題化合物(34.2 mg，19%)。1 H NMR：(DMSO－d6 )δ(ppm)：10.00(s,1H),8.44(s,1H),7.96(s,1H),7.57－7.65(m,3H),7.49(t,J＝7.6 Hz,2H),7.36－7.43(m,2H),7.31－7.35(m,1H),4.59(s,2H),4.23(s,2H),3.74(s,3H),3.11－3.20(m,2H),2.37(t,J＝7.2 Hz,2H),1.60－1.71(m,2H),1.45－1.55(m,2H),1.32－1.43(m,2H).LRMS(ESI)：(計算值)460.5：(實驗值)461.3(MH)＋ 。 Step 5: (E)-N-(2-Aminophenyl)-4-(2-(4-(2,4-dionethiathiazolidine-5-yl)methyl)phenoxy)B Oxy )benzamide (353 ) is dissolved in CH 3 CN with acid 357 (141 mg, 0.37 mmol), benzene-1,2-diamine (40 mg, 0.37 mmol) and BOP (161 mg, 0.37 mmol). (5 mL). Triethylamine (0.73 mmol, 203 μL) was added and the reaction was stirred at room temperature for 18 hours. The solvent was removed under reduced pressure and EtOAcqqqqqqqqqqqq 1 H NMR: (DMSO-d 6 ) δ (ppm): 10.00 (s, 1H), 8.44 (s, 1H), 7.96 (s, 1H), 7.57-7.65 (m, 3H), 7.49 (t, J) =7.6 Hz, 2H), 7.36-7.43 (m, 2H), 7.31-7.35 (m, 1H), 4.59 (s, 2H), 4.23 (s, 2H), 3.74 (s, 3H), 3.11-3.20 ( m, 2H), 2.37 (t, J = 7.2 Hz, 2H), 1.60-1.71 (m, 2H), 1.45-1.55 (m, 2H), 1.32-1.43 (m, 2H). LRMS (ESI): ( Calculated value) 460.5: (experimental value) 461.3 (MH) + .
N－(2－胺基苯基)－4－(2－(4－((2,4－二酮基噻唑烷－5－基)甲基)苯氧基)乙氧基)苯甲醯胺(358)步驟1：4－(2－(4－((2,4－二酮基噻唑烷－5－基)甲基)苯氧基)乙氧基)苯甲酸甲酯(359) 向356(流程67)(672 mg，1.68 mmol)之1,4－二噁烷(10 mL)溶液中加入10% Pd/C(2.3 g，2.18 mmol)。在室溫下氫氣氛下攪拌所得混合物2天，使其經矽藻土墊過濾且減壓濃縮產生359(379 mg，56%)。LRMS(ESI)：(計算值)401.4；(實驗值)424.2(M＋Na)＋ 。1 H NMR：(CDCl3 )δ(ppm)：8.15(bs,1H),7.99(d,J＝8.8 Hz,2H),7.16(d,J＝8.8 Hz,2H),6.96(d,J＝8.8 Hz,2H),6.90(d,J＝8.8 Hz,2H),4.52(dd,J＝9.2,4.0 Hz,1H),4.38－4.37(m,2H),4.32－4.32(m,2H),3.89(s,3H),3.45(dd,J＝14.1,4.0 Hz,1H),3.14(dd,J＝14.0,9.2 Hz,1H)。 N-(2-Aminophenyl)-4-(2-(4-((2,4-dione)thiazolidin-5-yl)methyl)phenoxy)ethoxy)benzamide (358) Step 1: 4-(2-(4-((2,4-diketothiazolidin-5-yl)methyl)phenoxy)ethoxy)benzoic acid methyl ester (359) to 356 (Scheme 67) (672 mg, 1.68 mmol) in 1,4-dioxane (10 mL) was added 10% Pd / C (2.3 g, 2.18 mmol). The resulting mixture was stirred at room temperature under a hydrogen atmosphere for 2 days, filtered over Celite pad and concentrated under vacuo to yield 359 (379 mg, 56%). LRMS (ESI) :( calc) 401.4; (Found) 424.2 (M + Na) + . 1 H NMR: (CDCl 3 ) δ (ppm): 8.15 (bs, 1H), 7.99 (d, J = 8.8 Hz, 2H), 7.16 (d, J = 8.8 Hz, 2H), 6.96 (d, J = 8.8 Hz, 2H), 6.90 (d, J = 8.8 Hz, 2H), 4.52 (dd, J = 9.2, 4.0 Hz, 1H), 4.38-4.37 (m, 2H), 4.32-4.32 (m, 2H), 3.89 (s, 3H), 3.45 (dd, J = 14.1, 4.0 Hz, 1H), 3.14 (dd, J = 14.0, 9.2 Hz, 1H).
步驟2：4－(2－(4－((2,4－二酮基噻唑烷－5－基)甲基)苯氧基)乙氧基)苯甲酸(360) 向甲酯359(872 mg，2.17 mmol)之冰AcOH(30 mL)溶液中加入濃HCl(10 mL)。將混合物在120℃下加熱3小時。減壓移除溶劑產生標題化合物(833 mg，99%)。1 H NMR：(DMSO－d6 )δ(ppm)：12.61(bs,1H),12.01(s,1H),7.91(d,J＝8.8 Hz,2H),7.19(d,J＝8.8 Hz,2H),7.08(d,J＝8.8 Hz,2H),6.95(d,J＝8.8 Hz,2H),4.91(dd,J＝9.2,4.4 Hz,1H),4.40－4.42(m,2H),4.33－4.36(m,2H),3.32(d,J＝4.0 Hz,1H),3.10(dd,J＝14.0,9.6 Hz,1H).LRMS(ESI)：(計算值)387.4；(實驗值)386.2(M－H)－ 。 Step 2: 4-(2-(4-((2,4-Diketothiazolidin-5-yl)methyl)phenoxy)ethoxy)benzoic acid (360) to methyl ester 359 (872 mg Concentrated HCl (10 mL) was added to a solution of 2.17 mmol) of ice AcOH (30 mL). The mixture was heated at 120 ° C for 3 hours. The solvent was removed under reduced pressure to give the title compound ( </ 1 H NMR: (DMSO-d 6 ) δ (ppm): 12.61 (bs, 1H), 12.01 (s, 1H), 7.91 (d, J = 8.8 Hz, 2H), 7.19 (d, J = 8.8 Hz, 2H), 7.08 (d, J = 8.8 Hz, 2H), 6.95 (d, J = 8.8 Hz, 2H), 4.91 (dd, J = 9.2, 4.4 Hz, 1H), 4.40-4.42 (m, 2H), 4.33-4.36 (m, 2H), 3.32 (d, J = 4.0 Hz, 1H), 3.10 (dd, J = 14.0, 9.6 Hz, 1H). LRMS (ESI): (calculated) 387.4; (experimental value) 386.2 (M-H) - .
步驟3：N－(2－胺基苯基)－4－(2－(4－((2,4－二酮基噻唑烷－5－基)甲基)苯基)乙氧基)苯甲醯胺(358) 依照用於合成化合物393所述之程序(步驟5，流程67，實例191)但以酸360代替酸357獲得呈米色固體之標題化合物(57 mg，產率33%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.34(s,1H),7.74－7.78(m,3H),6.93－6.98(m,3H),6.88(dJ＝8.8 Hz,2H),6.73－6.78(m,3H),6.57(dd,J＝8.0,1.2 Hz,1H),6.38(dt,J＝8.0,1.2 Hz,1H),4.68(dd,J＝8.8,4.4 Hz,2H),4.19－4.21(m,2H),3.11(d,J＝4.4 Hz,1H),2.88(dd,J＝14.0,9.2 Hz,1H).LRMS(ESI)：計算值477.4；(實驗值)478.4(MH)＋ 。 Step 3: N-(2-Aminophenyl)-4-(2-(4-((2,4-dionethiathiazol-5-yl)methyl)phenyl)ethoxy)benzene Amides (358) in accordance with the procedure described for the synthesis of compound 393 (step 5, process 67, example 191) but the acid 360 in place of the acid 357 title compound was obtained of a beige solid (57 mg, 33% yield). 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.34 (s, 1H), 7.74-7.78 (m, 3H), 6.93-6.98 (m, 3H), 6.88 (dJ = 8.8 Hz, 2H), 6.73-6.78(m,3H), 6.57 (dd, J=8.0, 1.2 Hz, 1H), 6.38 (dt, J=8.0, 1.2 Hz, 1H), 4.68 (dd, J=8.8, 4.4 Hz, 2H) , 4.19-4.21 (m, 2H), 3.11 (d, J = 4.4 Hz, 1H), 2.88 (dd, J = 14.0, 9.2 Hz, 1H). LRMS (ESI): calculated 477.4; (experimental value) 478.4 (MH) + .
(S)－N－(2－胺基苯基)－4－(3－(吡啶－3－基胺基)吡咯啶－1－基)苯甲醯胺(361)步驟1：N－對硝基苯磺酸－3－吡啶(362)： 向2－胺基吡啶(3.03 g，32.2 mmol)之經攪拌THF(15mL)溶液中相繼加入DCM(30 mL)、氯化4－硝基苯磺醯基(1.50 g，68.7mmol)及Et3 N(9.88 mL，70.9 mmol)。溶液變為橙色且形成沉澱物。將該懸浮液在室溫下攪拌1小時，減壓蒸發溶劑且將固體殘餘物懸浮於甲醇(200 mL)中。向該懸浮液加入極大過量(>10當量)之甲醇鈉，在50℃下攪拌該混合物3小時，以HCl 1 N(2 mL)中止且在80℃下減壓濃縮至體積變為~50 mL。又以1 N HCl酸化該濃縮溶液至中性pH。過濾收集所形成之沉澱物產生標題化合物(7.67 g，產率85%)。1 H NMR(DMSO－d6 )δ(ppm)：10.88(s,1H),8.36(d,J＝9.0 Hz,2H),8.28(dd,J＝6.1,1.4 Hz,1H),8.27(d,J＝2.5 Hz,1H),7.50(ddd,J＝8.4,2.7,1.6 Hz,1H),7.30(ddd,J＝8.2,4.7,0.8 Hz,1H).m/z：280.1(MH＋ )。 (S)-N-(2-Aminophenyl)-4-(3-(pyridin-3-ylamino)pyrrolidin-1-yl)benzamide (361) Step 1: N-p-nitro Toluenesulfonic acid-3-pyridine (362): To a stirred solution of 2-aminopyridine (3.03 g, 32.2 mmol) in THF (15 mL), DCM (30 mL) Mercapto (1.50 g, 68.7 mmol) and Et 3 N (9.88 mL, 70.9 mmol). The solution turned orange and a precipitate formed. The suspension was stirred at room temperature for 1 hr. A very large excess (>10 equivalents) of sodium methoxide was added to the suspension, and the mixture was stirred at 50 ° C for 3 hours, quenched with HCl 1 N (2 mL) and concentrated under reduced pressure at 80 ° C to a volume of ~50 mL. . The concentrated solution was again acidified to neutral pH with 1 N HCl. The precipitate formed was collected by filtration to give the title compound ( 7.67 g, yield: 85%). 1 H NMR (DMSO-d 6 ) δ (ppm): 10.88 (s, 1H), 8.36 (d, J = 9.0 Hz, 2H), 8.28 (dd, J = 6.1, 1.4 Hz, 1H), 8.27 (d) , J=2.5 Hz, 1H), 7.50 (ddd, J=8.4, 2.7, 1.6 Hz, 1H), 7.30 (ddd, J=8.2, 4.7, 0.8 Hz, 1H).m/z: 280.1 (MH + ) .
步驟2：4－((R)－3－羥基吡咯啶－1－基)苯甲酸第三丁酯(363)： 向4－氟苯甲酸第三丁酯(2.17 g，11.0 mmol)及(R)－(＋)－3－吡咯啶醇(1.00 g，11.5 mmol)之DMSO(8 mL)溶液中加入碳酸鉀(1.53 g，11.0 mmol)。在130℃下攪拌該混合物18小時且將其趁熱傾入至正攪拌之水(100 mL)中。過濾收集所得米色沉澱物且在120℃下將其乾燥1.5小時產生標題化合物(2.64 g，產率91%)。1 H NMR(丙酮－d6 )δ(ppm)：7.78(d,J＝9.0 Hz,2H),6.54(d,J＝8.8 Hz,2H),4.58(bs,1H),4.15(bs,1H),3.55(dd,J＝10.36,4.7 Hz,1H),3.49(t,J＝6.8 Hz,1H),3.42(td,J＝9.2,2.3 Hz,1H),3.28(d,J＝10.8 Hz,1H),2.21－2.10(m,1H),2.09－2.03(m,1H),1.56(s,9H).m/z：520.3(MH＋ )。 Step 2: 4 - ((R) -3- hydroxy-pyrrolidin-1-yl) benzoic acid tert-butyl ester (363): To 4-fluoro-benzoic acid tert-butyl ester (2.17 g, 11.0 mmol) and (R To a solution of -(+)-3-pyrrolidone (1.00 g, 11.5 mmol) in DMSO (8 mL). The mixture was stirred at 130 ° C for 18 hours and poured into hot water (100 mL) while stirring. The resulting beige precipitate was filtered and dried <RTI ID=0.0> 1 H NMR (acetone-d 6 ) δ (ppm): 7.78 (d, J = 9.0 Hz, 2H), 6.54 (d, J = 8.8 Hz, 2H), 4.58 (bs, 1H), 4.15 (bs, 1H) ), 3.55 (dd, J = 10.36, 4.7 Hz, 1H), 3.49 (t, J = 6.8 Hz, 1H), 3.42 (td, J = 9.2, 2.3 Hz, 1H), 3.28 (d, J = 10.8 Hz) , 1H), 2.21-2.10 (m, 1H), 2.09-2.03 (m, 1H), 1.56 (s, 9H).m/z: 520.3 (MH + ).
步驟3：4－((S)－3－N－對硝基苯磺酸(吡啶－3－基胺基)吡咯啶－1－基)苯甲酸第三丁酯(364) 向化合物362(6.00 g，21.5 mmol)之THF(100 mL)溶液中相繼加入原醇363(5.66 g，21.5 mmol)、三苯膦(6.76 g，25.8 mmol)及偶氮二甲酸二乙酯(4.06 mL,25.8 mmol)。在室溫下攪拌該混合物18小時且在真空中移除溶劑。使用EtOAc/Hex(40：60)作為溶離劑由急驟層析法純化殘餘物產生標題化合物(4.68 g，產率42%)。l H NMR(DMSO－d6 )d(ppm)：8.58(dd,J＝4.7,1.4 Hz,1H),8.48(d,J＝8.0 Hz,2H),8.38(d,J＝2.0 Hz,1H),8.13(d,J＝9.0,2H),7.72(d,J＝9.0 Hz,2H),7.61(ddd,J＝8.0,2.5,1.6 Hz,1H),7.39(dd,J＝8.2,4.9 Hz,1H),6.43(d,J＝9.0 Hz,2H),5.17(五重峰,J＝8.2 Hz,1H),3.77(dd,J＝10.4,7.2 Hz,1H),3.36(dd,J＝10.4,6.7 Hz,1H),3.26(dd,J＝15.1,7.8 Hz,1H),3.06(td,J＝12.3,3.3 Hz,1H),2.43－2.38(m,1H),2.02－1.94(m,1H),1.55(s,9H).m/z：525.3(MH＋ )。 Step 3: 4-((S)-3-N-p-Nitrobenzenesulfonic acid (pyridin-3-ylamino)pyrrolidin-1-yl)benzoic acid tert-butyl ester (364) to compound 362 (6.00 g, 21.5 mmol) in THF (100 mL) was added with hexanes 363 (5.66 g, 21.5 mmol), triphenylphosphine (6.76 g, 25.8 mmol) and diethyl azodicarboxylate (4.06 mL, 25.8 mmol) ). The mixture was stirred at room temperature for 18 hours and the solvent was removed in vacuo. The residue was purified by flash chromatography eluting elut elut elut elut elut elut elut l H NMR (DMSO-d 6 )d (ppm): 8.58 (dd, J = 4.7, 1.4 Hz, 1H), 8.48 (d, J = 8.0 Hz, 2H), 8.38 (d, J = 2.0 Hz, 1H) ), 8.13 (d, J = 9.0, 2H), 7.72 (d, J = 9.0 Hz, 2H), 7.61 (ddd, J = 8.0, 2.5, 1.6 Hz, 1H), 7.39 (dd, J = 8.2, 4.9 Hz, 1H), 6.43 (d, J = 9.0 Hz, 2H), 5.17 (five peaks, J = 8.2 Hz, 1H), 3.77 (dd, J = 10.4, 7.2 Hz, 1H), 3.36 (dd, J =10.4, 6.7 Hz, 1H), 3.26 (dd, J = 15.1, 7.8 Hz, 1H), 3.06 (td, J = 12.3, 3.3 Hz, 1H), 2.43-2.38 (m, 1H), 2.02-1.94 ( m, 1H), 1.55 (s , 9H) .m / z: 525.3 (MH +).
步驟4：4－((S)－3－(吡啶－3－基胺基)吡咯啶－1－基)苯甲酸第三丁酯(365) 向硝基化合物364(4.68 g，8.92 mmol)之DMF(45mL)溶液中相繼加入氫氧化鋰(1.31 g，31.2 mmol)及硫代乙醇酸(930μL，13.4 mmol)。在室溫下攪拌該混合物3天，在80℃下真空移除溶劑且使殘餘物在EtOAc與H2 O之間分溶。收集有機層且以HCl 1 N萃取之。收集酸性層且以飽和NaHCO3 溶液中和之。將形成之白色沉澱物以EtOAc進行萃取。以鹽水洗滌EtOAc溶液，使其經MgSO4 乾燥且在真空下濃縮產生呈白色固體之標題化合物(1.65 g，產率54%)。1 H NMR：(丙酮－d6 )δ(ppm)：8.08(d,J＝2.2 Hz,1H),7.86(d,J＝4.3 Hz,1H),7.79(d,J＝9.0 Hz,2H),7.09(dd,J＝8.2,4.3 Hz,1H),7.05(ddd,J＝8.2,2.7,1.6 Hz,1H),6.57(d,J＝8.8 Hz,2H),5.54(d,J＝6.5 Hz,1H),4.32(六重峰,J＝5.3 Hz,1H),3.78(dd,J＝10.2,5.9 Hz,1H),3.56(dd,J＝17.0,7.2 Hz,1H),3.47(td,J＝8.0,5.1 Hz,1H),3.31(dd,J＝10.2,3.9 Hz,1H),2.44(六重峰,J＝7.8 Hz,1H),2.13(六重峰,J＝5.1 Hz,1H),1.56(s,9H).m/z：340.3(MH＋ )。 Step 4: 4-((S)-3-(pyridin-3-ylamino)pyrrolidin-1-yl)benzoic acid tert-butyl ester (365) to nitro compound 364 (4.68 g, 8.92 mmol) Lithium hydroxide (1.31 g, 31.2 mmol) and thioglycolic acid (930 μL, 13.4 mmol) were successively added to a solution of DMF (45 mL). The mixture was stirred at room temperature for 3 days, the solvent was removed in vacuo at 80 deg.] C and the residue was partitioned between EtOAc and H 2 O. The organic layer was collected and extracted with HCl 1 N. Acidic layer was collected and NaHCO 3 saturated solution and the. The white precipitate which formed was extracted with EtOAc. With EtOAc solution was washed with brine, it was dried over MgSO 4 and concentrated under vacuo to give the title compound as a white solid (1.65 g, 54% yield). 1 H NMR: (acetone-d 6 ) δ (ppm): 8.08 (d, J = 2.2 Hz, 1H), 7.86 (d, J = 4.3 Hz, 1H), 7.79 (d, J = 9.0 Hz, 2H) , 7.09 (dd, J = 8.2, 4.3 Hz, 1H), 7.05 (ddd, J = 8.2, 2.7, 1.6 Hz, 1H), 6.57 (d, J = 8.8 Hz, 2H), 5.54 (d, J = 6.5 Hz, 1H), 4.32 (sixfold, J = 5.3 Hz, 1H), 3.78 (dd, J = 10.2, 5.9 Hz, 1H), 3.56 (dd, J = 17.0, 7.2 Hz, 1H), 3.47 (td , J = 8.0, 5.1 Hz, 1H), 3.31 (dd, J = 10.2, 3.9 Hz, 1H), 2.44 (six-peak, J = 7.8 Hz, 1H), 2.13 (six-peak, J = 5.1 Hz, 1H), 1.56 (s, 9H). m/z: 340.3 (MH + ).
步驟5：(S)－N－(2－胺基苯基)－4－(3－(吡啶－3－基胺基)吡咯啶－1－基)苯甲醯胺(361) 向化合物365(19 mg，0.56 mmol)之DCM 500μL懸浮液中加入三氟乙酸(200μL)。使溶液在50℃下回流3小時且在真空中濃縮產生白色固體。將該材料溶解於DMF(500μL)中且以Et3 N(16μL，0.118 mmol)及BOP(30 mg，0.067 mmol)進行處理。將反應混合物攪拌10分鐘且添加1,2－苯二胺(7 mg，0.061 mmol)及另一份Et3 N(23μL，0.168 mmol)。在室溫下攪拌該混合物2小時且在80℃真空下移除DMF。使殘餘物在EtOAc與H2 O之間分溶。收集且以1 N HCl萃取有機層並以飽和NaHCO3 中和之。將形成之沉澱物以EtOAc進行萃取，經MgSO4 乾燥且在真空下濃縮。用MeOH/CHCl3 (7：93)作為溶離劑由急驟層析法純化殘餘物產生標題化合物(11 mg，產率52%)。1 H NMR：(CD3 OD)δ(ppm)：7.97(d,J＝2.7 Hz,1H),7.86(d,J＝8.8 Hz,2H),7.78(dd,J＝4.7,1.0 Hz,1H),7.18－7.10(m,3H),7.05(td,J＝7.4,0.6 Hz,1H),6.89(dd,J＝7.8,1.2 Hz,1H),6.76(td,J＝7.4,1.4 Hz,1H),6.64(d,J＝8.8 Hz,2H),4.25(五重峰,J＝4.9 Hz,1H),3.77(dd,J＝10.2,6.1 Hz,1H),3.57(dd,J＝17.0,7.0 Hz,1H),3.49(td,J＝8.0,5.3 Hz,1H),3.29(q,J＝6.7 Hz,1H),2.41(六重峰,J＝7.2 Hz,1H),2.10(六重峰,J＝4.9 Hz,1H).m/z：372.4(MH＋ )。 Step 5: (S)-N-(2-Aminophenyl)-4-(3-(pyridin-3-ylamino)pyrrolidin-1-yl)benzamide (361) to compound 365 ( Trifluoroacetic acid (200 μL) was added to a 500 μL suspension of 19 mg, 0.56 mmol) of DCM. The solution was refluxed at 50 °C for 3 hours and concentrated in vacuo to give a white solid. This material was dissolved in DMF (500 μL) and treated with Et 3 N (16 μL, 0.118 mmol) and BOP (30 mg, 0.067 mmol). The reaction mixture was stirred for 10 min and 1,2-phenylenediamine (7 mg, 0.061 mmol) and another portion of Et 3 N (23μL, 0.168 mmol ). The mixture was stirred at room temperature for 2 hours and DMF was removed under vacuum at 80 °C. The residue was partitioned between EtOAc and H 2 O. The organic layer was collected and extracted with 1 N HCl and saturated NaHCO 3 and to the. The precipitate formed was extracted in EtOAc, 4 and concentrated under vacuo and dried over MgSO. Eluent to give the title compound (11 mg, 52% yield) was purified by a flash chromatography of the residue using MeOH / CHCl 3 (7:93) as the. 1 H NMR: (CD 3 OD) δ (ppm): 7.97 (d, J = 2.7 Hz, 1H), 7.86 (d, J = 8.8 Hz, 2H), 7.78 (dd, J = 4.7, 1.0 Hz, 1H ), 7.18-7.10 (m, 3H), 7.05 (td, J = 7.4, 0.6 Hz, 1H), 6.89 (dd, J = 7.8, 1.2 Hz, 1H), 6.76 (td, J = 7.4, 1.4 Hz, 1H), 6.64 (d, J = 8.8 Hz, 2H), 4.25 (five peaks, J = 4.9 Hz, 1H), 3.77 (dd, J = 10.2, 6.1 Hz, 1H), 3.57 (dd, J = 17.0) , 7.0 Hz, 1H), 3.49 (td, J = 8.0, 5.3 Hz, 1H), 3.29 (q, J = 6.7 Hz, 1H), 2.41 (six-peak, J = 7.2 Hz, 1H), 2.10 (six Heavy peak, J = 4.9 Hz, 1H). m/z: 372.4 (MH + ).
(R)－N－(2－胺基苯基)－4－(3－(吡啶－3－基胺基)吡咯啶－1－基)苯甲醯胺(366) 依照與流程69實例193所述相同之程序但以(S)－(－)－3－吡咯啶醇代替(R)－(＋)－3－吡咯啶醇獲得標題化合物(108 mg，產率21%)。1 H NMR：(DMSO－d6 )δ(ppm)：9.34(s,1H),8.00(d,J＝2.3 Hz,1H),7.84(d,J＝8.8 Hz,2H),7.77(dd,J＝4.7,1.2 Hz,1H),7.12(d,J＝7.6 Hz,1H),7.08(dd,J＝8.0,4.5 Hz,1H),6.99－6.97(m,1H),6.92(t,J＝7.8 Hz,1H),6.75(d,J＝7.8 Hz,1H),6.60－6.56(m,3H),6.17(d,J＝6.8 Hz,1H),4.81(s,2H),4.19－4.17(m,1H),3.71(dd,J＝10.2,6.5 Hz,1H),3.53－3.47(m,1H),3.42－3.38(m,1H),3.18(dd,J＝10.4,4.1 Hz,1H),2.32(六重峰,J＝6.3 Hz,1H),1.99(六重峰,J＝4.7 Hz,1H).m/z：374.2(MH＋ )。 (R)-N-(2-Aminophenyl)-4-(3-(pyridin-3-ylamino)pyrrolidin-1-yl)benzamide (366) according to Example 193 of Scheme 69 The same procedure was used but the title compound (108 mg, yield 21%) was obtained from (S)-(-)-3-pyrrolidin. 1 H NMR: (DMSO-d 6 ) δ (ppm): 9.34 (s, 1H), 8.00 (d, J = 2.3 Hz, 1H), 7.84 (d, J = 8.8 Hz, 2H), 7.77 (dd, J=4.7, 1.2 Hz, 1H), 7.12 (d, J=7.6 Hz, 1H), 7.08 (dd, J=8.0, 4.5 Hz, 1H), 6.99-6.97 (m, 1H), 6.92 (t, J = 7.8 Hz, 1H), 6.75 (d, J = 7.8 Hz, 1H), 6.60-6.56 (m, 3H), 6.17 (d, J = 6.8 Hz, 1H), 4.81 (s, 2H), 4.19 - 4.17 (m, 1H), 3.71 (dd, J = 10.2, 6.5 Hz, 1H), 3.53-3.47 (m, 1H), 3.42-3.38 (m, 1H), 3.18 (dd, J = 10.4, 4.1 Hz, 1H) ), 2.32 (sixfold, J = 6.3 Hz, 1H), 1.99 (six-peak, J = 4.7 Hz, 1H). m/z: 374.2 (MH + ).
(S)－N－(2－胺基苯基)－4－(3－(吡啶－3－基氧基)吡咯啶－1－基)苯甲醯胺(367) 依照與流程69實例193所述相同之程序但略過步驟1及4且以3－羥基吡啶代替化合物362獲得標題化合物(24 mg，產率44%)。1 H NMR：(丙酮－d6 )δ(ppm)：8.88(s,1H),8.31(d,J＝2.9 Hz,1H),8.19(d,J＝4.7 Hz,1H),7.93(d,J＝8.8 Hz,2H),7.41(ddd,J＝8.4,2.9,1.4 Hz,1H),7.31(ddd,J＝8.4,4.5,0.6 Hz,1H),7.26(d,J＝7.8 Hz,1H),6.97(td,J＝7.8,1.4 Hz,1H),6.85(dd,J＝8.0,1.4 Hz,1H),6.66(d,J＝9.0 Hz,2H),6.65(td,J＝8.0,1.4 Hz,1H),5.34－5.32(m,1H),4.62(s,2H),3.84(dd,J＝11.3,4.7 Hz,1H),3.61－3.56(m,3H),2.50－2.34(m,2H).m/z：375.2(MH＋ )。 (S)-N-(2-Aminophenyl)-4-(3-(pyridin-3-yloxy)pyrrolidin-1-yl)benzamide (367) according to Example 193 of Scheme 69 The same procedure was followed but the steps 1 and 4 were skipped and the title compound (24 mg, yield 44%) was obtained. 1 H NMR: (acetone-d 6 ) δ (ppm): 8.88 (s, 1H), 8.31 (d, J = 2.9 Hz, 1H), 8.19 (d, J = 4.7 Hz, 1H), 7.93 (d, J = 8.8 Hz, 2H), 7.41 (ddd, J = 8.4, 2.9, 1.4 Hz, 1H), 7.31 (ddd, J = 8.4, 4.5, 0.6 Hz, 1H), 7.26 (d, J = 7.8 Hz, 1H) ), 6.97 (td, J = 7.8, 1.4 Hz, 1H), 6.85 (dd, J = 8.0, 1.4 Hz, 1H), 6.66 (d, J = 9.0 Hz, 2H), 6.65 (td, J = 8.0, 1.4 Hz, 1H), 5.34-5.32 (m, 1H), 4.62 (s, 2H), 3.84 (dd, J = 11.3, 4.7 Hz, 1H), 3.61-3.56 (m, 3H), 2.50-2.34 (m , 2H).m/z: 375.2 (MH + ).
(R)－N－(2－胺基苯基)－4－(3－(吡啶－3－基氧基)吡咯啶－1－基)苯甲醯胺(368) 依照與流程69實例193所述相同之程序但略過步驟1及4且以3－羥基吡啶代替化合物362且以(S)－(－)－3－吡咯啶醇代替(R)－(＋)－3－吡咯啶醇獲得標題化合物(14 mg，產率12%)。1 H NMR：(丙酮－d6 )δ(ppm)：8.85(s,1H),8.31(d,J＝2.9 Hz,1H),8.19(dd,J＝4.5,1.2 Hz,1H),7.93(d,J＝8.8 Hz,2H),7.42(ddd,J＝8.4,2.9,1.4 Hz,1H),7.31(ddd,J＝8.4,4.7,0.8 Hz,1H),7.26(d,J＝7.8,1.6 Hz,1H),6.97(td,J＝7.2,1.6 Hz,1H),6.85(dd,J＝7.8,1.2 Hz,1H),6.68(d,J＝8.8 Hz,6.66(td,J＝7.6,1.4 Hz,1H),3.56－5.33(m,1H),4.60(bs,2H),3.86(dd,J＝11.3,4.7 Hz,1H),3.62－3.57(m,3H),2.50－2.35(m,2H).m/z：375.2(MH＋ )。 (R)-N-(2-Aminophenyl)-4-(3-(pyridin-3-yloxy)pyrrolidin-1-yl)benzamide (368) according to Example 193 of Scheme 69 The same procedure was followed but steps 1 and 4 were skipped and 3-hydroxypyridine was substituted for compound 362 and (S)-(-)-3-pyrrolidin was substituted for (R)-(+)-3-pyrrolidin. The title compound (14 mg, yield 12%). 1 H NMR: (acetone-d 6 ) δ (ppm): 8.85 (s, 1H), 8.31 (d, J = 2.9 Hz, 1H), 8.19 (dd, J = 4.5, 1.2 Hz, 1H), 7.93 ( d, J = 8.8 Hz, 2H), 7.42 (ddd, J = 8.4, 2.9, 1.4 Hz, 1H), 7.31 (ddd, J = 8.4, 4.7, 0.8 Hz, 1H), 7.26 (d, J = 7.8, 1.6 Hz, 1H), 6.97 (td, J = 7.2, 1.6 Hz, 1H), 6.85 (dd, J = 7.8, 1.2 Hz, 1H), 6.68 (d, J = 8.8 Hz, 6.66 (td, J = 7.6) , 1.4 Hz, 1H), 3.56-5.33 (m, 1H), 4.60 (bs, 2H), 3.86 (dd, J = 11.3, 4.7 Hz, 1H), 3.62-3.57 (m, 3H), 2.50-2.35 ( m, 2H). m/z: 375.2 (MH + ).
(S)－N－(2－胺基苯基)－4－(3－(苯胺基)吡咯啶－1－基)苯甲醯胺(369) 依照與流程69實例193所述相同之程序但以苯胺代替化合物3－胺基吡啶獲得標題化合物(7 mg，產率16%)。1 H NMR：(丙酮－d6 )δ(ppm)：8.84(s,1H),7.91(d,J＝8.8 Hz,2H),7.25(dd,J＝7.8,1.2 Hz,1H),7.12(t,J＝7.2 Hz,2H),6.96(dt,J＝8.0,1.4 Hz,1H),6.85(d,J＝8.0 Hz,1H),6.71(dd,J＝8.8,1.0 Hz,2H),6.66(t,J＝7.8 Hz,1H),6.62(d,J＝8.8 Hz,2H),5.26(d,J＝7.6 Hz,1H),4.60(bs,1H),4.30(五重峰,J＝5.3 Hz,1H),3.79(dd,J＝10.0 Hz,1H),3.57(q,J＝9.6 Hz,1H),3.50－3.45(m,1H),3.30(dd,J＝10.2,3.9 Hz,1H),2.42(六重峰,J＝6.8 Hz,1H).m/z：373.1(MH＋ )。 (S)-N-(2-Aminophenyl)-4-(3-(anilino)pyrrolidin-1-yl)benzamide (369) according to the same procedure as described in Example 193 of Scheme 69 but The title compound (7 mg, yield 16%) was obtained from phenylamine. 1 H NMR: (acetone-d 6 ) δ (ppm): 8.84 (s, 1H), 7.91 (d, J = 8.8 Hz, 2H), 7.25 (dd, J = 7.8, 1.2 Hz, 1H), 7.12 ( t, J = 7.2 Hz, 2H), 6.96 (dt, J = 8.0, 1.4 Hz, 1H), 6.85 (d, J = 8.0 Hz, 1H), 6.71 (dd, J = 8.8, 1.0 Hz, 2H), 6.66 (t, J = 7.8 Hz, 1H), 6.62 (d, J = 8.8 Hz, 2H), 5.26 (d, J = 7.6 Hz, 1H), 4.60 (bs, 1H), 4.30 (five peaks, J =5.3 Hz, 1H), 3.79 (dd, J = 10.0 Hz, 1H), 3.57 (q, J = 9.6 Hz, 1H), 3.50-3.45 (m, 1H), 3.30 (dd, J = 10.2, 3.9 Hz , 1H), 2.42 (sixfold, J = 6.8 Hz, 1H). m/z: 373.1 (MH + ).
(R)－N－(2－胺基苯基)－4－(3－(苯胺基)吡咯啶－1－基)苯甲醯胺(370) 依照與流程69實例193所述相同之程序但以苯胺代替化合物3－胺基吡啶且以(S)－3－吡咯啶醇代替(R)－(＋)－3－吡咯啶醇獲得標題化合物(22 mg，產率23%)。1 H NMR(CDCl3 )δ7.79(m,2H),7.2－7.4(m,2H),7.05(s,1H),6.8(m,3H)6.65(m,2H),6.53(m,2H),4.24(br.s.,1H),3.9(m,2H),3.73(m,1H),3.26(m,1H),2.37(m,1H),2.09(m,1H)m/z：373.3(MH＋ )。 (R)-N-(2-Aminophenyl)-4-(3-(anilino)pyrrolidin-1-yl)benzamide (370) was subjected to the same procedure as described in Example 193 of Scheme 69. The title compound (22 mg, yield 23%) was obtained from the titled compound (2-). 1 H NMR (CDCl 3 ) δ 7.79 (m, 2H), 7.2-7.4 (m, 2H), 7.05 (s, 1H), 6.8 (m, 3H) 6.65 (m, 2H), 6.53 (m, 2H) ), 4.24 (br.s., 1H), 3.9 (m, 2H), 3.73 (m, 1H), 3.26 (m, 1H), 2.37 (m, 1H), 2.09 (m, 1H) m/z: 373.3 (MH + ).
(S)－N－(2－胺基苯基)－4－(3－苯氧基吡咯啶－1－基)苯甲醯胺(371) 依照與流程69實例193所述相同之程序但略過步驟1及4且以苯酚代替化合物362獲得標題化合物(50 mg，產率33%)。1 H NMR(CDCl3 )δ7.79(μ,3H),7.3(m,3H),7.03(m,1H),6.96(m,1H),6.90(d,2H,J＝8.8 Hz),6.80(m,2H),6.54(d,J＝8.8 Hz,2H),5.08(br.s.,1H),3.71(dd,J＝4.7 Hz,J＝11.0 Hz,1H),3.6(m,3H),2.41(m,1H),2.31(m,1H)m/z：374.2(MH＋ )。 (S)-N-(2-Aminophenyl)-4-(3-phenoxypyrrolidin-1-yl)benzamide (371) according to the same procedure as described in Example 193 of Scheme 69. The title compound (50 mg, yield 33%) was obtained after sub.1. 1 H NMR (CDCl 3 ) δ 7.79 (μ, 3H), 7.3 (m, 3H), 7.03 (m, 1H), 6.96 (m, 1H), 6.90 (d, 2H, J = 8.8 Hz), 6.80 (m, 2H), 6.54 (d, J = 8.8 Hz, 2H), 5.08 (br.s., 1H), 3.71 (dd, J = 4.7 Hz, J = 11.0 Hz, 1H), 3.6 (m, 3H) ), 2.41 (m, 1H), 2.31 (m, 1H) m/z: 374.2 (MH + ).
(S)－甲基－4－(1－(4－(2－胺基苯胺甲醯基)苯基)吡咯啶－3－基氧基)苯甲酸酯(372) 依照與流程69實例193所述相同之程序但略過步驟1及4且以4－羥基苯甲酸甲酯代替化合物362獲得標題化合物(143 mg,產率42%)。1 H NMR(CDCl3 )δ8.0(m,2H),7.81(m,2H),7.72(s,1H),7.25(m,1H),7.06(m,1H),6.91(m,2H),6.84(m,2H),6.59(m,2H),5.16(br.s.,1H),3.9(s,3H),3.78(m,1H),3.60(m,3H),2.4(m,2H)m/z：432.4(MH＋ )。 (S)-Methyl-4-(1-(4-(2-aminoanilinocarbyl)phenyl)pyrrolidin-3-yloxy)benzoate (372) according to Example 69 with Scheme 69 The same procedure was followed except that steps 1 and 4 were repeated and the title compound (143 mg, yield 42%) was obtained. 1 H NMR (CDCl 3 ) δ 8.0 (m, 2H), 7.81 (m, 2H), 7.72 (s, 1H), 7.25 (m, 1H), 7.06 (m, 1H), 6.91 (m, 2H) , 6.84 (m, 2H), 6.59 (m, 2H), 5.16 (br.s., 1H), 3.9 (s, 3H), 3.78 (m, 1H), 3.60 (m, 3H), 2.4 (m, 2H) m/z: 432.4 (MH + ).
(S)－4－(1－(4－(2－胺基苯基胺甲醯基)苯基)吡咯啶－3－基氧基)苯甲酸(373) 在室溫下將溶於THF/水/MeOH之1：1：1混合物(6 mL)中之372(100 mg，0.23 mmol)及KOH(100 mg，1.78 mmol)溶液攪拌5天。將反應混合物濃縮且使其在水(5 mL)與***(5 mL)之間分溶。丟棄有機層且將含水相以1 M HCl溶液酸化至pH＝6且以EtOAc萃取。使萃取物經Na2 SO4 乾燥，過濾且加以濃縮。以MeOH－DCM為溶離劑(5至20% MeOH梯度之MeOH)由急驟層析法純化該殘餘物產生標題化合物(20 mg，產率21%)。1 H NMR(DMSO)δ9.34(a,1H),7.85(m,4H),7.11(d,1H,J＝7.8 Hz),7.00(d,J＝8.4 Hz,2H),6.92(d,2H,J＝7.7 Hz),6.75(d,J＝8.0 Hz,2H),6.6(m,3H),5.26(br.s.,1H),3.75(m,1H),[3.34 DMSO,4H],2.44(m,1H),2.31(m,1H)m/z：418.4(MH＋ )。 (S)-4-(1-(4-(2-Aminophenylaminocarbamimidyl)phenyl)pyrrolidin-3-yloxy)benzoic acid (373) will be dissolved in THF at room temperature. A solution of 372 (100 mg, 0.23 mmol) and KOH (100 mg, 1.78 mmol) in a 1 : 1 mixture of water/MeOH (6 mL) was stirred for 5 days. The reaction mixture was concentrated and partitioned between water (5 mL) and diethyl ether (5 mL). The organic layer was taken and the aqueous phase was acidified to pH = 6 with EtOAc EtOAc. The extract was dried over Na 2 SO 4, filtered and concentrated. The residue was purified by flash chromatography eluting elut elut elut elut elut elut elut elut 1 H NMR (DMSO) δ 9.34 (a, 1H), 7.85 (m, 4H), 7.11 (d, 1H, J = 7.8 Hz), 7.00 (d, J = 8.4 Hz, 2H), 6.92 (d, 2H, J=7.7 Hz), 6.75 (d, J=8.0 Hz, 2H), 6.6 (m, 3H), 5.26 (br.s., 1H), 3.75 (m, 1H), [3.34 DMSO, 4H] , 2.44 (m, 1H), 2.31 (m, 1 H) m/z: 418.4 (MH + ).
(S)－N－(2－胺基苯基)－4－(3－(3,4,5－三甲氧基苯氧基)吡咯啶－1－基)苯甲醯胺(374) 依照與流程69實例193所述相同之程序但略過步驟1及4且以3,4,5－三甲氧基苯酚代替化合物362獲得標題化合物(30 mg，21%)。1 H NMR(CDCl3 )δ7.79(d,2H,J＝8.8 Hz),7.73(s,1H),7.26(d,1H,J＝7.4 Hz),7.05(t,J＝7.7 Hz,1H),6.81(d,2H,J＝7.7 Hz),6.57(d,J＝8.7 Hz,2H),6.14(s,2H),5.04(br.s.,1H),3.88(s,6H),3.80(s,3H),3.71(dd,J＝4.7 Hz,J＝11.0 Hz,1H),3.6(m,3H),2.41(m,1H),2.31(m,1H)m/z：464.4(MH＋ )。 (S)-N-(2-aminophenyl)-4-(3-(3,4,5-trimethoxyphenoxy)pyrrolidin-1-yl)benzamide (374) according to The same procedure as described in Example 193 of Scheme 69, but substituting steps 1 and 4 and substituting 3,4,5-trimethoxyphenol for compound 362 afforded the title compound (30 mg, 21%). 1 H NMR (CDCl 3 ) δ 7.79 (d, 2H, J = 8.8 Hz), 7.73 (s, 1H), 7.26 (d, 1H, J = 7.4 Hz), 7.05 (t, J = 7.7 Hz, 1H) ), 6.81 (d, 2H, J = 7.7 Hz), 6.57 (d, J = 8.7 Hz, 2H), 6.14 (s, 2H), 5.04 (br.s., 1H), 3.88 (s, 6H), 3.80(s,3H), 3.71 (dd, J=4.7 Hz, J=11.0 Hz, 1H), 3.6 (m, 3H), 2.41 (m, 1H), 2.31 (m, 1H) m/z: 464.4 ( MH + ).
(S)－N－(2－胺基苯基)－4－(3－(苯幷[d][1,3]二氧雜環戊烯－5－基氧)吡咯啶－1－基)苯甲醯胺(375) 依照與流程69實例193所述相同之程序但略過步驟1及4且以苯幷[d][1,3]二氧雜環戊烯－5－醇代替化合物362獲得標題化合物(31 mg，15%)。1 H NMR(CDCl3 )δ8.95(s,1H),7.87(d,2H,J＝8.0 Hz),7.80(d,J＝8.7 Hz,1H),7.67(s,1H),7.48(s,.5H)7.04(m,1H),6.83(m,1H),6.71(m,1H),6.57(d,2H),6.48(s,1H),6.33(m,1H),5.93(s,2H),4.96(br.s.,1H),3.67(m,1H),3.57(m,3H),2.36(m,1H),2.26(m,1H)m/z：418.2(MH＋ )。 (S)-N-(2-Aminophenyl)-4-(3-(benzoquinone[d][1,3]dioxol-5-yloxy)pyrrolidin-1-yl) Benzoamide (375) followed the same procedure as described in Example 193 of Scheme 69 but skipped steps 1 and 4 and replaced the compound 362 with phenylhydrazine [d][1,3]dioxol-5-ol. The title compound was obtained (31 mg, 15%). 1 H NMR (CDCl 3 ) δ 8.95 (s, 1H), 7.87 (d, 2H, J = 8.0 Hz), 7.80 (d, J = 8.7 Hz, 1H), 7.67 (s, 1H), 7.48 (s) , .5H) 7.04 (m, 1H), 6.83 (m, 1H), 6.71 (m, 1H), 6.57 (d, 2H), 6.48 (s, 1H), 6.33 (m, 1H), 5.93 (s, 2H), 4.96 (br.s., 1H), 3.67 (m, 1H), 3.57 (m, 3H), 2.36 (m, 1H), 2.26 (m, 1H) m/z: 418.2 (MH + ).
(S)－N－(2－胺基苯基)－4－(3－(4－苯氧基苯氧基)吡咯啶－1－基)苯甲醯胺(376) 依照與流程69實例193所述相同之程序但略過步驟1及4且以4－苯氧基苯酚代替化合物362獲得標題化合物。1 H NMR(CDCl3 )δ7.81(m,2H),7.70(s,1H),7.67(s,.5H),7.2－7.4(m,4H)6.8－7.2(m,10H),6.6(m,2H),5.05(br.s.,1H),3.6(m,1H),3.5(m,3H),2.41(m,1H),2.32(m,1H)m/z：466.4(MH＋ )。 (S)-N-(2-Aminophenyl)-4-(3-(4-phenoxyphenoxy)pyrrolidin-1-yl)benzamide (376) according to Example 69 with Scheme 69 The same procedure was followed except that steps 1 and 4 were skipped and the title compound was obtained by substituting 4-phenoxyphenol for compound 362. 1 H NMR (CDCl 3 ) δ 7.81 (m, 2H), 7.70 (s, 1H), 7.67 (s, .5H), 7.2-7.4 (m, 4H) 6.8-7.2 (m, 10H), 6.6 ( m, 2H), 5.05 (br.s., 1H), 3.6 (m, 1H), 3.5 (m, 3H), 2.41 (m, 1H), 2.32 (m, 1H) m/z: 466.4 (MH + ).
(S)－N－(2－胺基苯基)－4－(3－(4－硝基苯氧基)吡咯啶－1－基)苯甲醯胺(377) 依照與流程69實例193所述相同之程序但略過步驟1及4且以4－硝基苯酚代替化合物362獲得標題化合物(12 mg,6%)。1 H NMR(CDCl3 )δ8.12(d,2H,J＝9.1 Hz),7.72(d,J＝8.8 Hz,2H),7.18(d,J＝7.3 Hz,1H),6.97(t,1H,J＝7.7 Hz),6.87(d,J＝9.1 Hz,2H),6.50(d,2H,J＝8.6 Hz),5.09(br.s.,1H),3.71(dd,J＝4.5 Hz,J＝11.3 Hz,1H),3.6(m,3H),2.3(m,2H)m/z：419.1(MH＋ )。 (S)-N-(2-Aminophenyl)-4-(3-(4-nitrophenoxy)pyrrolidin-1-yl)benzamide (377) according to Example 193 of Scheme 69 The same procedure was followed but the steps 1 and 4 were skipped and the title compound (12 mg, 6%) was obtained. 1 H NMR (CDCl 3 ) δ 8.12 (d, 2H, J = 9.1 Hz), 7.72 (d, J = 8.8 Hz, 2H), 7.18 (d, J = 7.3 Hz, 1H), 6.97 (t, 1H) , J = 7.7 Hz), 6.87 (d, J = 9.1 Hz, 2H), 6.50 (d, 2H, J = 8.6 Hz), 5.09 (br.s., 1H), 3.71 (dd, J = 4.5 Hz, J = 11.3 Hz, 1H), 3.6 (m, 3H), 2.3 (m, 2H) m/z: 419.1 (MH + ).
(S)－N－(2－胺基苯基)－4－(3－(吡啶－2－基硫基)吡咯啶－1－基)苯甲醯胺(378) 依照與流程69實例193所述相同之程序但略過步驟1及4且以吡啶－2－硫醇代替化合物362獲得標題化合物(22 mg，28%)。1 H NMR(CDCl3 )δ8.44(μ,1H),7.78(d,J＝8.8 Hz,2H),7.69(s,1H),7.49(t,1H,J＝7.4 Hz),7.27(m,1H),7.18(d,1H,J＝8.0 Hz),7.0－7.1(m,2H),6.82(d,7.8 Hz,2H),6.55(d,J＝9.0 Hz,2H),4.55(m,1H),3.9－4.0(m,3H),3.4－3.6(m,4H),2.6(m,1H),2.2(m,1H)m/z：391.0(MH＋ )。 (S)-N-(2-Aminophenyl)-4-(3-(pyridin-2-ylthio)pyrrolidin-1-yl)benzamide (378) according to Example 193 of Scheme 69 The same procedure was followed but the steps 1 and 4 were skipped and the title compound (22 mg, 28%) was obtained from pyridine-2-thiol. 1 H NMR (CDCl 3 ) δ 8.44 (μ, 1H), 7.78 (d, J = 8.8 Hz, 2H), 7.69 (s, 1H), 7.49 (t, 1H, J = 7.4 Hz), 7.27 (m) , 1H), 7.18 (d, 1H, J = 8.0 Hz), 7.0-7.1 (m, 2H), 6.82 (d, 7.8 Hz, 2H), 6.55 (d, J = 9.0 Hz, 2H), 4.55 (m) , 1H), 3.9-4.0 (m, 3H), 3.4-3.6 (m, 4H), 2.6 (m, 1H), 2.2 (m, 1H) m/z: 391.0 (MH + ).
N－(2－胺基苯基)－5－((4－(吡啶－2－基)嘧啶－2－基胺基)甲基)噻吩－2－甲醯胺(379)步驟1：2－(5－甲醯基噻吩－2－甲醯胺基)苯基胺基甲酸第三丁酯(380) 在室溫下將5－甲醯基噻吩－2－羧酸(350 mg，2.24 mmol)、2－胺基苯基胺基甲酸第三丁酯(467 mg，2.24 mmol)及三乙胺(470μL，340 mg，3.36 mmol)及BOP(1.1 g，2.68 mmol)之DMF(10 mL)溶液攪拌2小時。濃縮反應混合物且以己烷中之25% EtOAc作為溶離劑由急驟層析法加以純化產生260 mg(33%)標題化合物。1 H NMR(CDCl3 )δ9.95(s,1H),9.7(br.s,1H),7.84(d,1H,J＝8.0 Hz),7.74(m,2H),7.15(m,2H),6.72(s,1H),1.56(s,9H).LRMS：(計算值)346.1；(實驗值)369.1(M＋Na)。 N-(2-Aminophenyl)-5-((4-(pyridin-2-yl)pyrimidin-2-ylamino)methyl)thiophene-2-carboxamide (379) Step 1: 2 (5-Mercaptothiophene-2-carboxamido) phenylaminocarbamic acid tert- butyl ester (380) 5-Methylthiophene-2-carboxylic acid (350 mg, 2.24 mmol) at room temperature , 3-aminophenylaminocarbamic acid tert-butyl ester (467 mg, 2.24 mmol) and triethylamine (470 μL, 340 mg, 3.36 mmol) and BOP (1.1 g, 2.68 mmol) in DMF (10 mL) Stir for 2 hours. The reaction mixture was concentrated and purified by flash chromatography eluting elut elut elut 1 H NMR (CDCl 3 ) δ 9.95 (s, 1H), 9.7 (br.s, 1H), 7.84 (d, 1H, J = 8.0 Hz), 7.74 (m, 2H), 7.15 (m, 2H) , 6.72 (s, 1H), 1.56 (s, 9H). LRMS: (calc.) 346.1; (found) 369.1 (M+Na).
步驟2：2－(5－((4－(吡啶－2－基)嘧啶－2－基胺基)甲基)噻吩－2－甲醯胺基)苯基胺基甲酸第三丁酯(381) 以NaBH(OAc)3 (22 mg，1 mmol)處理醛380(260 mg，0.75 mmol)、4－(吡啶－3－基)嘧並－2－胺(85 mg，0.5 mmol)及乙酸(100 μL)之DCE(2 mL)溶液且將所得混合物在室溫下攪拌隔夜。接著由添加飽和NaHCO3 (5 mL)中止且以EtOAc萃取含水相。將有機萃取物經Na2 SO4 乾燥，過濾且進行濃縮提供粗產物，用80－20% EtOAc－己烷混合物作為溶離劑由急驟層析法純化該粗產物產生標題化合物(35 mg，產率14%)。LRMS：(計算值)502.2；(實驗值)503.4(M＋H1 )。 Step 2: 2-(5-((4-(Pyridin-2-yl)pyrimidin-2-ylamino)methyl)thiophene-2-carboxamido) phenylaminocarbamic acid tert-butyl ester (381 ) to NaBH (OAc) 3 (22 mg , 1 mmol) treated aldehyde 380 (260 mg, 0.75 mmol) , 4- ( pyridin-3-yl) pyrimidin-2-amine and (85 mg, 0.5 mmol) and acetic acid ( 100 μL) of DCE (2 mL) solution and the mixture was stirred overnight at room temperature. And then with a saturated NaHCO 3 (5 mL) The aqueous suspension was extracted with EtOAc. The organic extract was dried over Na 2 SO 4, filtered and concentrated to provide the crude product as eluent to give the title compound (35 mg purified by flash chromatography of the crude product was 80-20% EtOAc- hexane mixture as yield 14%). LRMS :( calc) 502.2; (Found) 503.4 (M + H 1) .
N－(2－胺基苯基)－5－((4－(吡啶－2－基)嘧啶－2－基胺基)甲基)噻吩－2－甲醯胺(379) 在室溫下將溶於DCM與TFA之1：1混合物(4 mL)中之381 (35 mg，0.07 mmol)溶液攪拌30分鐘。濃縮反應混合物產生固體，以***濕磨該固體產生呈TFA鹽之標題化合物(26 mg，產率75%)。1 H NMR(MeOH－d4 )δ9.31(s,1H),8.68(s,1H),8.66(s,1H),8.43(d,J＝5.1 Hz,1H),7.77(d,J＝3.9 Hz,1H),7.68(m,1H),7.1－7.5(m,6H).LRMS：(計算值)402.2；(實驗值)403.3(M＋H1 )。 N-(2-Aminophenyl)-5-((4-(pyridin-2-yl)pyrimidin-2-ylamino)methyl)thiophene-2-carboxamide (379) will be at room temperature A solution of 381 (35 mg, 0.07 mmol) in 1:1 mixture of DCM and TFA (4 mL) was stirred for 30 min. The reaction mixture was concentrated to give a crystal crystal crystal crystal crystal crystal crystal crystal crystal 1 H NMR (MeOH-d 4 ) δ 9.31 (s, 1H), 8.68 (s, 1H), 8.66 (s, 1H), 8.43 (d, J = 5.1 Hz, 1H), 7.77 (d, J = 3.9 Hz, 1H), 7.68 ( m, 1H), 7.1-7.5 (m, 6H) .LRMS :( calc) 402.2; (Found) 403.3 (m + H 1) .
N－(2－胺基苯基)－4－((3－(6－甲氧基吡啶－3－基)－1H－吡唑－5－基胺基)甲基)苯甲醯胺(382)步驟1：3－(6－甲氧基吡啶－3－基)－1H－吡唑－5－胺(383) 在－78℃下以BuLi之己烷(7.2 mL，17.96 mmol)2.5 M溶液處理THF(20 mL)中之MeCN(940μL，736 mg，17.96 mmol)且將反應混合物在相同溫度下攪拌30分鐘，將其在－78℃下以6－甲氧基煙酸甲酯(2 g，11.96 mmol)之THF(10 mL)溶液處理且在室溫下再攪拌2小時。接著由添加水(10 mL)及1 M HCl溶液(10 mL)使其中止。將所得混合物在真空下濃縮，使殘餘物與溶於EtOH(30 mL)中之單水合肼(5 mL)混合，回流2小時，冷卻且減壓濃縮產生固體，用DCM中之10% MeOH作為溶離劑由急驟層析法純化該固體產生標題化合物(720 mg，產率32%)。1 H NMR(MeOH－d4 )δ8.39(s,1H),7.89(d,J＝8.6 Hz,1H),6.80(d,J＝8.6 Hz,1H),5.85(s,1H),3.91(s,3H).LRMS：(計算值)190.1；(實驗值)191.1(M＋H1 )。 N-(2-Aminophenyl)-4-((3-(6-methoxypyridin-3-yl)-1H-pyrazol-5-ylamino)methyl)benzamide (382 Step 1: 3-(6-Methoxypyridin-3-yl)-1H-pyrazole-5-amine (383) at -78 ° C as a BuLi hexane (7.2 mL, 17.96 mmol) 2.5 M solution Treatment of MeCN (940 μL, 736 mg, 17.96 mmol) in THF (20 mL) and the reaction mixture was stirred at the same temperature for 30 min and then taken at -78 ° C with methyl 6-methoxynicotinate (2 g A solution of 11.96 mmol) in THF (10 mL) was stirred at room temperature for 2 hr. This was followed by addition of water (10 mL) and 1 M HCl solution (10 mL). The mixture was concentrated in vacuo and EtOAc EtOAc EtOAc m. The lysing agent was purified by flash chromatography to give the title compound ( s. 1 H NMR (MeOH-d 4 ) δ 8.39 (s, 1H), 7.89 (d, J = 8.6 Hz, 1H), 6.80 (d, J = 8.6 Hz, 1H), 5.85 (s, 1H), 3.91 (s, 3H) .LRMS :( calc) 190.1; (Found) 191.1 (M + H 1) .
步驟2：4－((3－(6－甲氧基吡啶－3－基)－1H－吡唑－5－基胺基)甲基)苯甲酸甲酯(384) 在室溫下將胺383(720 mg，3.79 mmol)、4－甲醯基苯甲酸甲酯(745 mg，4.54 mmol)及Bu2 SnCl2 (230 mg，0.76 mmol)之無水THF(5 mL)溶液攪拌2小時。接著以PhSiH3 (514μL，451 mg，4.17 mmol)處理該溶液且在室溫下再攪拌四小時。由添加MeOH中止反應混合物且將其劇烈攪拌45分鐘。接著將其在真空下濃縮且使用己烷中梯度為50－100%之EtOAc由急驟層析法純化該殘餘物產生標題化合物(672 mg，產率52%)。LRMS：(計算值)338.1；(實驗值)339.2(M＋H1 )步驟3：4－((3－(6－甲氧基吡啶－3－基)－1H－吡唑－5－基胺基)甲基)苯甲酸(385) 在室溫下將THF、MeOH及水之1：1：1之混合物(9 mL)中之384(672 mg，1.99 mmol)及KOH(300 mg，5.35 mmol)溶液攪拌隔夜。由添加1 M HCl將反應混合物酸化至pH＝4且在真空下濃縮之。以水濕磨殘餘物且過濾收集固體且進行乾燥產生標題化合物(640 mg，產率99%)。LRMS：(計算值)324.1；(實驗值)325.2(M＋H1 )步驟4：N－(2－胺基苯基)－4－((3－(6－甲氧基吡啶－3－基)－1H－吡唑－5－基胺基)甲基)苯甲醯胺(382) 相繼以Et3 N(831μL，603 mg，5.96 mmol)、EDC(571 mg，2.98 mmol)、HOBT(334 mg，2.18 mmol)及1,2－苯二胺(429 mg，3.97 mmol)處理385(640 mg，1.97 mmol)之MeCN(10 mL)溶液且將其攪拌隔夜。濃縮反應混合物且使其在DCM(15 mL)與飽和NH4 Cl(15 mL)之間分溶。收集有機相，將其以Na2 SO4 乾燥，過濾及濃縮。使用DCM中梯度為3－15%之MeOH由急驟層析法純化所得固體產生標題化合物(113 mg，產率14%)。1 H NMR(DMSO－d6 )δ9.65(s,1H),8.43(s,1H),7.92(m,3H),7.48(d,J＝8.0 Hz,2H),7.16(d,J＝7.4 Hz,1H),6.97(t,J＝7.6 Hz,1H),6.8l(t,J＝8.2 Hz,2H),6.63(t,J＝7.4 Hz,1H),5.85(s,1H),4.34(s,2H),3.85(s,3H).LRMS：(計算值)414.2；(實驗值)415.3(M＋H1 )。 Step 2: 4 - ((3- (6-methoxypyridin-3-yl) lH-pyrazol-5-yl) methyl) benzoate (384) A solution of amine 383 (720 mg, 3.79 mmol), 4- methyl acyl acid methyl ester (745 mg, 4.54 mmol) and Bu 2 SnCl 2 (230 mg, 0.76 mmol) of anhydrous THF (5 mL) was stirred for 2 hours. Followed by PhSiH 3 (514μL, 451 mg, 4.17 mmol) and treating the solution was stirred at room temperature for another four hours. The reaction mixture was quenched by the addition of MeOH and stirred vigorously for 45 min. The residue was purified by EtOAc EtOAcqqqqq LRMS: (calculated) 338.1; (experimental value) 339.2 (M+H 1 ) Step 3: 4-((3-(6-methoxypyridin-3-yl)-1H-pyrazol-5-ylamino) Methyl)benzoic acid (385) 384 (672 mg, 1.99 mmol) and KOH (300 mg, 5.35 mmol) in a 1:1:1 mixture of THF, MeOH and water (9 mL ) at room temperature Stir overnight. The reaction mixture was acidified to pH = 4 by addition of 1 M HCl and concentrated in vacuo. The residue was triturated with EtOAc (EtOAc)EtOAc. LRMS :( calc) 324.1; (Found) 325.2 (M + H 1) Step 4: N- (2- aminophenyl) -4 - ((3- (6-methoxypyridin-3-yl) - 1H-pyrazol-5-ylamino)methyl)benzamide (382) successively as Et 3 N (831 μL, 603 mg, 5.96 mmol), EDC (571 mg, 2.98 mmol), HOBT (334 mg, 2.18 mmol) and 1,2-phenylenediamine (429 mg, 3.97 mmol) were treated with 385 (640 mg, 1.97 mmol) MeCN (10 mL) and stirred overnight. The reaction mixture was concentrated and allowed to DCM (15 mL) and partitioned between saturated NH 4 Cl (15 mL). Collect the organic phase, which was dried in Na 2 SO 4, filtered and concentrated. The resulting solid was purified by flash chromatography eluting elut elut elut elut elut elut 1 H NMR (DMSO-d 6 ) δ 9.65 (s, 1H), 8.43 (s, 1H), 7.92 (m, 3H), 7.48 (d, J = 8.0 Hz, 2H), 7.16 (d, J = 7.4 Hz, 1H), 6.97 (t, J = 7.6 Hz, 1H), 6.8l (t, J = 8.2 Hz, 2H), 6.63 (t, J = 7.4 Hz, 1H), 5.85 (s, 1H), 4.34 (s, 2H), 3.85 (s, 3H) .LRMS :( calc) 414.2; (Found) 415.3 (M + H 1) .
實例209：N－(2－胺基苯基)－4－((3－(吡啶－3－基)－1H－吡唑－5－基胺基)甲基)苯甲醯胺(386)Example 209: N-(2-Aminophenyl)-4-((3-(pyridin-3-yl)-1H-pyrazol-5-ylamino)methyl)benzamide (386)
根據流程71(實例208)自煙酸甲酯開始製備標題化合物。1 H NMR(MeOH－d4 )δ8.80(s,1H),8.43(d,J＝3.9 Hz,1H),8.04(m,2H),7.94(d,J＝8.2 Hz,2H),7.53(d,J＝8.2 Hz,1H),7.43(m,1H),7.15(d,J＝7.6 Hz,1H),7.05(t,J＝7.2 Hz,2H),6.88(d,J＝8.0 Hz,1H),6.75(t,J＝7.4 Hz,1H),5.94(s,1H),4.45(s,2H).LRMS：(計算值)384.2；(實驗值)385.2(M＋H1 )。The title compound was prepared starting from methyl nicotinate according to Scheme 71 (Example 208). 1 H NMR (MeOH-d 4 ) δ8.80 (s, 1H), 8.43 (d, J = 3.9 Hz, 1H), 8.04 (m, 2H), 7.94 (d, J = 8.2 Hz, 2H), 7.53 (d, J = 8.2 Hz, 1H), 7.43 (m, 1H), 7.15 (d, J = 7.6 Hz, 1H), 7.05 (t, J = 7.2 Hz, 2H), 6.88 (d, J = 8.0 Hz) , 1H), 6.75 (t, J = 7.4 Hz, 1H), 5.94 (s, 1H), 4.45 (s, 2H) .LRMS :( calc) 384.2; (Found) 385.2 (M + H 1) .
N－(2－胺基苯基)－4－((3－(3,4,5－三甲氧基苯基)－1H－吡唑－5－基胺基)甲基)苯甲醯胺(387)N-(2-Aminophenyl)-4-((3-(3,4,5-trimethoxyphenyl)-1H-pyrazol-5-ylamino)methyl)benzamide ( 387)
根據流程71(實例208)自3,4,5－三甲氧基苯甲酸甲酯開始製備標題化合物。1 H NMR(MeOH－d4 )δ7.92(d,J＝8.4 Hz,2H),7.53(d,J＝8.0 Hz,2H),7.16(d,J＝7.9 Hz,1H),7.06(t,J＝7.8 Hz,1H),6.93(s,2H),6.88(d,J＝8.0 Hz,1H),6.75(t,J＝7.6 Hz,1H),5.89(s,1H),4.45(s,2H),3.87(s,6H),3.77(s,3H).LRMS：(計算值)473.3；(實驗值)474.4(M＋H1 )。The title compound was prepared starting from methyl 3,4,5-trimethoxybenzoate according to Scheme 71 (Example 208). 1 H NMR (MeOH-d 4 ) δ 7.92 (d, J = 8.4 Hz, 2H), 7.53 (d, J = 8.0 Hz, 2H), 7.16 (d, J = 7.9 Hz, 1H), 7.06 (t , J = 7.8 Hz, 1H), 6.93 (s, 2H), 6.88 (d, J = 8.0 Hz, 1H), 6.75 (t, J = 7.6 Hz, 1H), 5.89 (s, 1H), 4.45 (s , 2H), 3.87 (s, 6H), 3.77 (s, 3H) .LRMS :( calc) 473.3; (Found) 474.4 (M + H 1) .
N－(2－胺基苯基)－4－((4－氯－3－(3,4,5－三甲氧基苯基)－1H－吡唑－5－基胺基)甲基)苯甲醯胺(388)N-(2-Aminophenyl)-4-((4-chloro-3-(3,4,5-trimethoxyphenyl)-1H-pyrazol-5-ylamino)methyl)benzene Formamide (388)
步驟1、2及3：4－((3－(3,4,5－三甲氧基苯基)－1H－吡唑－5－基胺基)甲基)苯甲酸(389) 根據流程71步驟1、2及3在第一步中使用3,4,5－三甲氧基苯甲酸甲酯代替6－甲氧基煙酸甲酯獲得標題化合物。LRMS：(計算值)383.1；(實驗值)384.2(M＋H1 )。 Steps 1, 2 and 3: 4-((3-(3,4,5-trimethoxyphenyl)-1H-pyrazol-5-ylamino)methyl)benzoic acid (389) according to Scheme 71 1, 2 and 3 In the first step, methyl 3,4,5-trimethoxybenzoate was used in place of methyl 6-methoxynicotinate to obtain the title compound. LRMS :( calc) 383.1; (Found) 384.2 (M + H 1) .
步驟4：N－(2－胺基苯基)－4－((4－氯－3－(3,4,5－三甲氧基苯基)－1H－吡唑－5－基胺基)甲基)苯甲醯胺(388) 在室溫下將溶於DCM與SOCl2 之1：1混合物(2 mL)中之389(30 mg，0.08 mmol)溶液攪拌30分鐘。濃縮反應混合物且以1,2－苯二胺(18 mg，0.16 mmol)之THF(2 mL)溶液處理並在室溫下攪拌15分鐘，減壓濃縮產生固體，經製備型HPLC(AQUASIL C－18柱；5μM；230 x 21.2 mm；溶離劑為水中之30－95% MeOH)純化該固體以產生標題化合物(8 mg，產率20%)。1 H NMR(MeOH－d4 )δ7.92(d,J＝8.3 Hz,2H),7.52(d,J＝8.2 Hz,2H),7.17(d,J＝7.6 Hz,1H),7.06(m,3H),6.89(d,J＝7.8 Hz,1H),6.75(t,J＝7.2 Hz,1H),4.55(s,2H),3.89(s,6H),3.80(s,3H).LRMS：(計算值)507.2；(實驗值)508.3(M＋H1 )。 Step 4: N-(2-Aminophenyl)-4-((4-chloro-3-(3,4,5-trimethoxyphenyl)-1H-pyrazol-5-ylamino) A yl) benzoyl-amine (388) was dissolved in DCM and 2 of the SOCl2 at room temperature for 1: 1 mixture (2 mL) in the 389 (30 mg, 0.08 mmol) was stirred for 30 minutes. The reaction mixture was concentrated and purified with EtOAc EtOAc EtOAcjjjjjjjj The title compound (8 mg, yield 20%) was purified eluting elute 1 H NMR (MeOH-d 4 ) δ 7.92 (d, J = 8.3 Hz, 2H), 7.52 (d, J = 8.2 Hz, 2H), 7.17 (d, J = 7.6 Hz, 1H), 7.06 (m) , 3H), 6.89 (d, J = 7.8 Hz, 1H), 6.75 (t, J = 7.2 Hz, 1H), 4.55 (s, 2H), 3.89 (s, 6H), 3.80 (s, 3H). LRMS : (calculated value) 507.2; (experimental value) 508.3 (M+H 1 ).
N－(2－胺基苯基)－4－((8－甲基－7－酮基－7,8－二氫吡啶幷[2,3－d]嘧啶－2－基胺基)甲基)苯甲醯胺(389)步驟1：2－(4－(第三丁氧基羰基－胺甲基)苯甲醯胺基)苯基胺基甲酸第三丁酯(391) 在室溫下相繼以EDC(930 mg，4.84 mmol)、HOBT(682 mg，4.46 mmol)及Et3 N(670μL，489 mg，4.84 mmol)處理4－((第三丁氧基羰基胺基)甲基)苯甲酸(1 g，3.98 mmol)之DMF(10 mL)溶液且攪拌隔夜。減壓濃縮反應混合物且使其在氯仿(10 mL)與水(10 mL)之間分溶。收集有機相，以1 M HCl(10 mL)及飽和NaHCO3 (10 mL)洗滌該有機相，使其乾燥，過濾及蒸發形成殘餘物，將用己烷中之30% EtOAc作為溶離劑由急驟層析法純化該殘餘物產生標題化合物(840 mg，51%)。LRMS：(計算值)441.2；(實驗值)442.2(M＋H1 )步驟2：4－(胺甲基)－N－(2－胺基苯基)苯甲醯胺(392) 在室溫下將DCM/TFA之2：1混合物(6 mL)中之391(840 mg，1.9 mmol)溶液攪拌2小時。在真空中濃縮反應混合物產生作為單TFA鹽及二TFA鹽之混合物之標題化合物(1.33 g，產率100%)。LRMS：(計算值)241.2；(實驗值)242.2(M＋H1 )。 N-(2-Aminophenyl)-4-((8-methyl-7-keto-7,8-dihydropyridinium[2,3-d]pyrimidin-2-ylamino)methyl Benzoguanamine (389) Step 1: 2-(4-(Tertiary Butoxycarbonyl-Aminemethyl)benzylideneamino)phenylaminocarboxylic acid tert-butyl ester (391) at room temperature Treatment of 4-((t-butoxycarbonylamino)methyl)benzene with EDC (930 mg, 4.84 mmol), HOBT (682 mg, 4.46 mmol) and Et 3 N (670 μL, 489 mg, 4.84 mmol) A solution of formic acid (1 g, 3.98 mmol) in DMF (10 mL) was stirred overnight. The reaction mixture was concentrated under reduced pressure and dried over EtOAc (EtOAc) The organic phase was collected, to 1 M HCl (10 mL) and saturated NaHCO 3 (10 mL) The organic phase was washed, dried, filtered and evaporated to form a residue with 30% EtOAc in hexanes as the eluent by flash The residue was purified by EtOAc EtOAcjjj: LRMS :( calc) 441.2; (Found) 442.2 (M + H 1) Step 2: 4- (aminomethyl) -N- (2- aminophenyl) benzoyl-amine (392) at the room temperature A solution of 391 (840 mg, 1.9 mmol) in a 2:1 mixture of DCM/TFA (6 mL) was stirred for 2 h. The reaction mixture was concentrated in vacuo to give title compound (l.l. LRMS :( calc) 241.2; (Found) 242.2 (M + H 1) .
步驟3：N－(2－胺基苯基)－4－((8－甲基－7－酮基－7,8－二氫吡啶幷[2,3－d]嘧啶－2－基胺基)甲基)苯甲醯胺(390)在室溫下將亞碸393(Barvian,M.等人J.Med.Chem.(2001 )44(6)；1016－1016)(166 mg，0.74 mmol)、雙胺392(535 mg，2.23 mmol)及三乙胺(620μL，4.46 mmol)之DME(3 mL)溶液攪拌3小時。濃縮該反應混合物且使其在EtOAc(5 mL)與水(5 mL)之間分溶。將有機相收集且相繼以NH4 Cl(5 mL)及NaHCO3 (5 mL)之飽和溶液洗滌，經Mg2 SO4 乾燥，過濾及濃縮產生殘餘物，以1：1之EtOAc/己烷溶液濕磨該殘餘物產生標題化合物(48 mg，產率16%)。1 H NMR(CDCl3 )δ3.62(s,3H),4.80(m,2H),6.42(d,J＝10 Hz,1H),6.85(d,J＝8 Hz,2H),7.10(m,1H),7.30(m,1H),7.50(m,2H),7.87(s,1H),7.897(m,2H),8.43(s,1H).LRMS：(計算值)400.0；(實驗值)401.0(M＋H1 )。Step 3: N-(2-Aminophenyl)-4-((8-methyl-7-keto-7,8-dihydropyridinium[2,3-d]pyrimidin-2-ylamino) Methyl)benzamide (390) Aachen 393 at room temperature (Barvian, M. et al. J. Med. Chem. ( 2001 ) 44(6); 1016-1016) (166 mg, 0.74 mmol) , a solution of dimethylamine 392 (535 mg, 2.23 mmol) and triethylamine (620 μL, 4.46 mmol) in DME (3 mL). The reaction mixture was concentrated and partitioned between EtOAc (5 mL)EtOAc. The organic phase was collected and dried successively washed with NH 4 Cl (5 mL) and NaHCO 3 (5 mL) the solution was saturated Mg 2 SO 4, filtered and concentrated to a residue, to 1: EtOAc 1 of / hexane solution The residue was triturated to give the title compound (48 mg, yield 16%). 1 H NMR (CDCl 3 ) δ 3.62 (s, 3H), 4.80 (m, 2H), 6.42 (d, J = 10 Hz, 1H), 6.85 (d, J = 8 Hz, 2H), 7.10 (m) , 1H), 7.30 (m, 1H), 7.50 (m, 2H), 7.87 (s, 1H), 7.879 (m, 2H), 8.43 (s, 1H). LRMS: (calculated) 400.0; ) 401.0 (M+H 1 ).
N－(2－胺基苯基)－4－((7－酮基－7,8－二氫吡啶幷[2,3－d]嘧啶－2－基胺基)甲基)苯甲醯胺(394)N-(2-Aminophenyl)-4-((7-keto-7,8-dihydropyridinium[2,3-d]pyrimidin-2-ylamino)methyl)benzamide (394)
以與實例212(流程72)相似之方法自藉由與亞碸393相似之文獻程序獲得之亞碸395開始製備標題化合物。1 H NMR(DMSO)δ4.60(s,2H),4.90(s,2H),6.10(d,J＝10 Hz,1H),6.55(t,J＝7 Hz,2H),6.75(m,1H),6.90(t,J＝7 Hz,2H),7.10(m,2H),7.40(m,2H),7.65(m,1H),7.90(m,1H),8.55(s,1H),9.69(s,1H).LRMS：(計算值)386.0；(實驗值)387.0(M＋H1 )。The title compound was prepared starting from hydrazine 395, obtained by a literature procedure similar to that of Aachen 393, in a similar manner to Example 212 (Scheme 72). 1 H NMR (DMSO) δ 4.60 (s, 2H), 4.90 (s, 2H), 6.10 (d, J = 10 Hz, 1H), 6.55 (t, J = 7 Hz, 2H), 6.75 (m, 1H), 6.90 (t, J = 7 Hz, 2H), 7.10 (m, 2H), 7.40 (m, 2H), 7.65 (m, 1H), 7.90 (m, 1H), 8.55 (s, 1H), 9.69 (s, 1H) .LRMS :( calc) 386.0; (Found) 387.0 (M + H 1) .
(S)－N－(1－(4－(2－胺基苯基胺甲醯基)苯基)吡咯啶－3－基)煙鹼醯胺(396)步驟1：(S)－第三丁基4－(3－胺基吡咯啶－1－基)苯甲酸酯(397) 用與步驟2流程69所述相同之程序類似於胺基酯363獲得標題化合物。1 H NMR(CDCl3 )δ7.83(d,J＝8.8 Hz,1H),6.46(d,J＝8.8 Hz,1H),3.75(m,1H),3.35－3.6(m,3H),3.06(dd,J＝4.7 Hz,J＝9.8 Hz,1H),2.26(m,1H),1.85(m,1H),1.57(s,9H),LRMS：(計算值)262.1；(實驗值)263.0(M＋H1 )。 (S)-N-(1-(4-(2-Aminophenylaminocarbamimidyl)phenyl)pyrrolidin-3-yl)nicotinium amide (396) Step 1: (S) - Third Butyl 4-(3-aminopyrrolidin-1-yl)benzoate (397) was obtained in the same procedure as the one obtained from the procedure 1 H NMR (CDCl 3 ) δ 7.83 (d, J = 8.8 Hz, 1H), 6.46 (d, J = 8.8 Hz, 1H), 3.75 (m, 1H), 3.35-3.6 (m, 3H), 3.06 (dd, J = 4.7 Hz, J = 9.8 Hz, 1H), 2.26 (m, 1H), 1.85 (m, 1H), 1.57 (s, 9H), LRMS: (calculated) 262.1; (experimental value) 263.0 (M+H 1 ).
步驟2：(S)－第三丁基4－(3－(煙鹼醯胺基)吡咯啶－1－基)苯甲酸酯(398) 在室溫下將397(100 mg，0.38 mmol)、Et3 N(160μL，1.14 mmol)及煙鹼醯基氯氫氯酸鹽(68 mg，0.38 mmol)之DCM(2 mL)溶液攪拌1小時且藉由加入NH4 Cl鹽飽和溶液(5 mL)中止之。分離有機相，使其經Na2 SO4 乾燥，過濾且濃縮以產生殘餘物，用DCM中之5% MeOH作為溶離劑由急驟層析法純化該殘餘物產生標題化合物(110 mg，產率79%)。LRMS：(計算值)367.2；(實驗值)。368.1(M＋H1 ). Step 2: (S)-Tertiary Butyl 4-(3-(nicotinoguanidino)pyrrolidin-1-yl)benzoate (398) 397 (100 mg, 0.38 mmol) at rt , Et 3 N (160μL, 1.14 mmol) and nicotine acyl chloride hydrochloride (68 mg, 0.38 mmol) of DCM (2 mL) was stirred for 1 hour and 4 Cl salt by addition of a saturated solution of NH (5 mL ) Suspended. The organic phase was separated, dried over it Na 2 SO 4, filtered and concentrated to give the residue with 5% MeOH in DCM as the eluent The residue was purified by flash chromatography to give the title compound (110 mg, yield 79 %). LRMS: (calculated) 367.2; (experimental value). 368.1 (M+H 1 ).
步驟3：(S)－4－(3－(煙鹼醯胺基)吡咯啶－1－基)苯甲酸(399) 用與步驟5流程28所述相同之程序類似於化合物117獲得呈單鹽及二鹽混合物之標題化合物。LRMS：(計算值)311.1；(實驗值)312.1(M＋H1 )。 Step 3: (S)-4-(3-(nicotinoguanamine)pyrrolidin-1-yl)benzoic acid (399) is obtained as a single salt using the same procedure as described for And the title compound of the di-salt mixture. LRMS :( calc) 311.1; (Found) 312.1 (M + H 1) .
步驟4：(S)－N－(1－4－(2－胺基苯胺甲醯基)苯基)吡咯啶－3－基)煙鹼醯胺(396) 在室溫下將399(93 mg，0.3 mmol)、苯二胺(65 mg，0.6 mmol)、EDC(86 mg，0.45 mmoI)、HOBT(53 mg，0.33 mmol)及Et3 N(125μL，91 mg，0.9 mmol)之乙腈(2 mL)溶液攪拌隔夜。濃縮反應混合物且以DCM中梯度為5－20%之MeOH作為溶離劑由急驟層析法純化殘餘物產生標題化合物(24 mg，產率16%)。1 H NMR(CDCl3 )δ8.97(s,1H),8.66(d,J＝4.9 Hz,1H),8.21(d,J＝3.9 Hz,1H),7.87(d,2H,J＝8.8 Hz),7.52(dd,J＝5.1 Hz,J＝8.0 Hz,1H),7.15(d,1H,J＝7.9 Hz),7.05(t,J＝8.1 Hz,1H),6.89(d,J＝7.6 Hz,1H),6.76(t,J＝7.3 Hz,1H),6.66(d,J＝9.0 Hz,2H),4.78(m,1H),3.80(dd,J＝6.7 Hz,J＝10.2 Hz,1H),3.61(m,1H),3.49(m,1H),3.41(m,1H),2.4(m,1H),2.2(m,1H).LRMS：(計算值)401.2；(實驗值)402.2(M＋H1 )。 Step 4: (S)-N-(1-4-(2-Aminoanilinecarbamoyl)phenyl)pyrrolidin-3-yl)nicotinamide (396) 399 (93 mg ) at room temperature , 0.3 mmol), phenylenediamine (65 mg, 0.6 mmol), EDC (86 mg, 0.45 mmoI), HOBT (53 mg, 0.33 mmol) and Et 3 N (125 μL, 91 mg, 0.9 mmol) of acetonitrile (2 The mL) solution was stirred overnight. The reaction mixture was concentrated and purified with EtOAcjjjjjjjjjj 1 H NMR (CDCl 3 ) δ 8.97 (s, 1H), 8.66 (d, J = 4.9 Hz, 1H), 8.21. (d, J = 3.9 Hz, 1H), 7.87 (d, 2H, J = 8.8 Hz ), 7.52 (dd, J = 5.1 Hz, J = 8.0 Hz, 1H), 7.15 (d, 1H, J = 7.9 Hz), 7.05 (t, J = 8.1 Hz, 1H), 6.89 (d, J = 7.6) Hz, 1H), 6.76 (t, J = 7.3 Hz, 1H), 6.66 (d, J = 9.0 Hz, 2H), 4.78 (m, 1H), 3.80 (dd, J = 6.7 Hz, J = 10.2 Hz, 1H), 3.61 (m, 1H), 3.49 (m, 1H), 3.41 (m, 1H), 2.4 (m, 1H), 2.2 (m, 1H). LRMS: (calculated) 401.2; (experimental value) 402.2 (M + H 1).
N－(2－胺基苯基)－4－((S)－3－((S)－吡啶－2－基亞磺醯基)吡咯啶－1－基)苯甲醯胺(400) 在室溫下將378(15 mg，0.04 mmol)及mCPBA(6 mg，0.04 mmol)之DCM(2 mL)溶液攪拌1小時。濃縮反應混合物且用DCM中之梯度EtOAc至5% MeOH作為溶離劑由急驟層析法純化殘餘物產生標題化合物(13 mg，產率80%)。1 H NMR(CDCl3 )δ1 H NMR(CDCl3 )δ8.63(m,1H),8.58(m,1H),7.6－8.0(m,10H),7.40(m,2H),7.25(m,1H),7.05(m,2H),6.85(m,3H),6.58(d,J＝8.8 Hz,2H),6.50(d,J＝11.1 Hz,2H),3.74.0(m,6H),3.2－3.5(m,4H),2.4－2.8(m,3H),2.95(m,1H).LRMS：(計算值)406.1；(實驗值)407.1(M＋H1 )。 N-(2-Aminophenyl)-4-((S)-3-((S)-pyridin-2-ylsulfinyl)pyrrolidin-1-yl)benzamide (400) A solution of 378 (15 mg, 0.04 mmol) and m.sub. The reaction mixture was concentrated with EtOAc EtOAcqqqqqqq 1 H NMR (CDCl 3) δ 1 H NMR (CDCl 3) δ8.63 (m, 1H), 8.58 (m, 1H), 7.6-8.0 (m, 10H), 7.40 (m, 2H), 7.25 (m , 1H), 7.05 (m, 2H), 6.85 (m, 3H), 6.58 (d, J = 8.8 Hz, 2H), 6.50 (d, J = 11.1 Hz, 2H), 3.74.0 (m, 6H) , 3.2-3.5 (m, 4H), 2.4-2.8 (m, 3H), 2.95 (m, 1H) .LRMS :( calc) 406.1; (Found) 407.1 (m + H 1) .
1.人類HDAC－1 檢定1.由桿狀病毒昆蟲細胞表現系統表現及純化的經選殖之重組性人類HDAC－1酶篩選HDAC抑制劑。為檢定去乙醯基酶，在37℃下以30μg經選殖之重組性hHDAC－1培育20,000 cpm[3 H]－代謝示蹤之經乙醯化組蛋白基質(M.Yoshida等人，J.Biol.Chem.265(28)：17174－17179(1990))10分鐘。藉由加入乙酸(0.04 M，最終濃度)及HCl(250 mM，最終濃度)停止反應。以乙酸乙酯萃取該混合物且藉由閃爍計數定量所釋放之[3 H]－乙酸。為研究抑制作用，在開始酶檢定之前在4℃下以化合物預培育該酶30分鐘。藉由以單獨化合物製作劑量反應曲線且測定產生50%最大抑制作用之抑制劑濃度來研究HDAC酶抑制劑之IC5 0 值。 1. Human HDAC-1 Assay 1. Screening for HDAC inhibitors by baculovirus insect cell expression system performance and purified colonized recombinant human HDAC-1 enzyme. To determine the deacetylase, 20,000 cpm [ 3 H]-metabolized acetaminophen histone matrix was incubated with 30 μg of selected recombinant hHDAC-1 at 37 ° C (M. Yoshida et al., J . Biol. Chem. 265 (28): 17174-17179 (1990)) 10 minutes. The reaction was stopped by the addition of acetic acid (0.04 M, final concentration) and HCl (250 mM, final concentration). The mixture was extracted with ethyl acetate and the released [ 3 H]-acetic acid was quantified by scintillation counting. To investigate inhibition, the enzyme was pre-incubated with the compound for 30 minutes at 4 °C prior to initiation of the enzyme assay. Produced by a separate dose of the compound inhibitor and a concentration-response curve to 50% of maximal inhibition assay to study the generation of HDAC inhibitors IC 5 0 value.
檢定2.亦用以下試驗計劃檢定本發明之化合物。在檢定中使用之緩衝劑為25 mM HEPES pH 8.0，137 mM NaCl、2.7 mM KCl、1 mM MgCl2 ，且基質為DMSO中之Boc－Lys(Ac)－AMC 50 mM儲備溶液。緩衝劑中該酶儲備溶液為4.08μg/mL。在室溫下將該等化合物(在緩衝劑中將DMSO中2μl稀釋至13μl以供轉移至檢定盤)與酶(20μl，4.08μg/ml)預培育(預培育體積35μl)10分鐘。在室溫下預培育該混合物5分鐘。藉由使溫度達到37℃且加入16μl基質開始該反應。總反應體積為50μl。20分鐘後藉由加入50μl顯色劑使反應停止，該顯色劑按Biomol(Fluor－de－Lys顯色劑，編號#KI－105)所指導來製備。在讀取前將盤在室溫下於暗處培育10分鐘(λE X ＝360 nm，λE m ＝470 nm，截止濾波在435 nm處)。Assay 2. The compounds of the invention were also assayed using the following test protocol. The buffer used in the assay was 25 mM HEPES pH 8.0, 137 mM NaCl, 2.7 mM KCl, 1 mM MgCl 2 and the matrix was a Boc-Lys(Ac)-AMC 50 mM stock solution in DMSO. The enzyme stock solution in the buffer was 4.08 μg/mL. The compounds (2 μl in DMSO were diluted to 13 μl in buffer for transfer to the assay plate) and the enzyme (20 μl, 4.08 μg/ml) were pre-incubated (pre-culture volume 35 μl) for 10 minutes at room temperature. The mixture was pre-incubated for 5 minutes at room temperature. The reaction was started by bringing the temperature to 37 ° C and adding 16 μl of the substrate. The total reaction volume was 50 μl. After 20 minutes, the reaction was stopped by the addition of 50 μl of a developer which was prepared under the guidance of Biomol (Fluor-de-Lys developer, #KI-105). The plates were incubated for 10 minutes at room temperature in the dark before reading (λ E X = 360 nm, λ E m = 470 nm, cutoff filter at 435 nm).
代表性化合物之IC5 0 值列於表14中。用檢定1量測如下化合物之HDAC活性：10c、13e、16d、26b、44、47、61a、61b、63、134、138及308。用檢定2量測如下化合物之HDAC活性：361、366、367、368、369、370、371、372、373、374、375、376、377及378。表14中，"a"表示0.1μM之活性，"b"表示1μM之活性，"c"表示5μM之活性且"d"表示>5μm之活性。對表14中H4－Ac T24 EC對MS－275之檢定，"u"表示小於1，"v"表示1且"w"表示大於1。對表14中之H3 Ac t24檢定，"x"表示1μM之活性，"y"表示10μM之活性且"z"表示20μM之活性。IC 50 values of representative compounds are shown in Table 14. The HDAC activities of the following compounds were determined by assay 1 : 10c, 13e, 16d, 26b, 44, 47, 61a, 61b, 63, 134, 138 and 308. The HDAC activities of the following compounds were determined by assay 2: 361, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377 and 378. In Table 14, "a" indicates Activity of 0.1 μM, "b" indicates 1 μM activity, "c" indicates Activity of 5 μM and "d" indicates activity of >5 μm. For the verification of H4-Ac T24 EC to MS-275 in Table 14, "u" means less than 1, "v" means 1 and "w" means greater than 1. For the H3 Ac t24 test in Table 14, "x" indicates 1 μM activity, "y" means 10 μM activity and "z" 20 μM activity.
2. MTT檢定 在處理化合物前一天將HCT116細胞(2000/孔)接入至96－孔組織培養盤中。將各種濃度之化合物加入細胞中。在37℃下於5% CO2 恆溫箱中培育細胞72小時。加入最終濃度為0.5 mg/ml之MTT(溴化3－[4,5－二甲基噻唑－4－基]－2,5二苯基四唑鹽，Σ)且將其與細胞培育4小時，再將一體積增溶緩衝劑(50% N,N－二甲基甲醯胺，20% SDS，pH 4.7)加至經培養之細胞上。隔夜培育後，藉由使用一MR700盤式讀取器(Dynatech Laboratories Inc.)以630 nM作為參照在570 nM處由比色分析讀取來量化經增溶之染料。根據相關細胞系之標準生長曲線將OD值轉換成細胞數。將令經溶劑處理之細胞數減少至50%的濃度確定為MTT IC5 0 。代表性化合物之IC5 0 值示於表14中。表14中，"a"表示0.1μM之活性，"b"表示1μM之活性且"c"表示5μM之活性。 2. MTT assay HCT116 cells (2000/well) were inserted into 96-well tissue culture plates one day prior to treatment of the compounds. Various concentrations of compounds are added to the cells. The cells were incubated for 72 hours at 37 ° C in a 5% CO 2 incubator. MTT (3-[4,5-dimethylthiazol-4-yl]-2,5-diphenyltetrazolium bromide, hydrazine) at a final concentration of 0.5 mg/ml was added and incubated with cells for 4 hours. A volume of solubilization buffer (50% N,N-dimethylformamide, 20% SDS, pH 4.7) was added to the cultured cells. After overnight incubation, the solubilized dye was quantified by colorimetric analysis reading at 570 nM using a MR700 disc reader (Dynatech Laboratories Inc.) with 630 nM as a reference. The OD value is converted to the number of cells according to the standard growth curve of the relevant cell line. The concentration at which the number of solvent-treated cells was reduced to 50% was determined as MTT IC 5 0 . IC 50 values of representative compounds are shown in Table 14. In Table 14, "a" indicates Activity of 0.1 μM, "b" indicates 1 μM activity and "c" 5 μM activity.
3.由免疫墨點在完整細胞中進行組蛋白H4乙醯化作用 將生長於培養物中之T24人類膀胱癌細胞與HDAC抑制劑一起培育16小時。如M.Yoshida等人(U.Biol.Chem.265(28)：17174－17179(1990))所述，在培養期後自細胞中萃取出組蛋白。將20 g總組蛋白載於SDS/PAGE上且轉移至硝化纖維膜。以特異於乙醯化組蛋白H－4(Upstate Biotech Inc.)之多株抗體，隨後以辣根過氧化物酶共軛二級抗體(Σ)探查該等膜。使用柯達(Kodak)底片(Eastman Kodak)執行增強化學發光(ECL)(Amersham)偵測。藉由密度測定法量化乙醯化H－4訊號。代表性資料顯示於表14中。資料表示為用本發明所示化合物有效減少50%乙醯化H－4訊號之濃度(EC5 0 )與一對照化合物MS－275之比率。若顯示之比率為1，則本發明之化合物與MS－275對照化合物同等有效。若該比率小於1，則本發明之化合物比MS－275對照化合物有效。可在Suzuki等人，J.Med.Chem.1999，第3001－3003頁中進一步發現關於MS－275化合物之資訊。 3. Histone H4 acetylation in intact cells by immunoblots T24 human bladder cancer cells grown in culture were incubated with HDAC inhibitors for 16 hours. Histones were extracted from the cells after the culture period as described by M. Yoshida et al. (U. Biol. Chem. 265(28): 17174-17179 (1990)). 20 g of total histone was loaded on SDS/PAGE and transferred to a nitrocellulose membrane. These membranes were probed with polyclonal antibodies specific for acetylated histone H-4 (Upstate Biotech Inc.) followed by horseradish peroxidase conjugated secondary antibodies (Σ). Enhanced chemiluminescence (ECL) (Amersham) detection was performed using Kodak negatives (Eastman Kodak). The acetylated H-4 signal was quantified by densitometry. Representative data are shown in Table 14. The data is expressed as the ratio of the concentration of the ethyl bromide H-4 signal (EC 5 0 ) to the control compound MS-275 which is effectively reduced by the compound of the present invention. If the ratio shown is 1, the compound of the invention is as effective as the MS-275 control compound. If the ratio is less than 1, the compound of the present invention is more effective than the MS-275 control compound. Information on MS-275 compounds can be further found in Suzuki et al., J. Med. Chem. 1999, pages 3001-3003.
4.組蛋白H3乙醯化作用檢定 將生長於培養物中之T24人類膀胱癌細胞與HDAC抑制劑一起培育16小時。藉由加入10μl Alamar Blue(BioSource,DAL1100)測定細胞存活率。將細胞以PBS洗滌一次且以預冷至－20℃ 10分鐘之甲醇固定。接著以PBS洗滌細胞兩次。以50μl PBS＋0.1% Triton X－100阻斷(block)固定細胞。以免－抗－乙醯基－H3(Upstate #06－599)作為初級抗體且接著以山羊－抗－兔－HRP(Σ#A－0545)作為二級抗體探查細胞。在加入Amplex－Red後藉由螢光計在Ex：550、Em：610、切斷：590(Auto PMT，15個讀數/孔)下讀取螢光度。以源自Alamar Blue之細胞存活率校正螢光訊號。資料以EC5 0 之方式顯示於表14中。MS－275最大乙醯化作用訊號(螢光度單位)量測為Em a x 。達到50% Em a x 之化合物濃度為EC5 0 。表14中，"x"表示1μM之活性，"y"表示10μM之活性且"z"表示20μM之活性。 4. Histone H3 acetylation assay T24 human bladder cancer cells grown in culture were incubated with HDAC inhibitor for 16 hours. Cell viability was determined by adding 10 μl of Alamar Blue (BioSource, DAL1100). The cells were washed once with PBS and fixed with methanol pre-cooled to -20 °C for 10 minutes. The cells were then washed twice with PBS. The cells were fixed with 50 μl PBS + 0.1% Triton X-100. The cells were probed with a non-anti-acetamido-H3 (Upstate #06-599) as a primary antibody and then goat-anti-rabbit-HRP (Σ#A-0545) as a secondary antibody. Fluorescence was read by a fluorometer at Ex: 550, Em: 610, cut: 590 (Auto PMT, 15 readings/well) after adding Amplex-Red. Fluorescence signals were corrected for cell viability derived from Alamar Blue. The data is shown in Table 14 in the form of EC 5 0 . The MS-275 maximum acetylation signal (fluorescence unit) is measured as E m a x . The concentration of the compound reaching 50% E m a x is EC 5 0 . In Table 14, "x" indicates 1 μM activity, "y" means 10 μM activity and "z" 20 μM activity.
在八至十週齡雌性BCD1小鼠(Taconic Labs,Great Barrington,NY)肋腹區域皮下注射2 x 106 個經預改良之HCT116人類結腸直腸癌細胞、SW48結腸癌細胞及A549肺癌細胞。藉由在裸小鼠之相同菌株內最少連續三次腫瘤移植來預改良該等細胞。隨後切除大約30 mg腫瘤片段且芬瑞尼(Forene)麻醉(Abbott Labs,Geneva,Switzerland)下將其皮下植入至小鼠體內之左肋腹區域中。當腫瘤平均體積達到100 mm3 時，藉由每日注射起始劑量為10 mg/kg之組蛋白去乙醯基酶抑制劑之DMSO溶液腹膜內處理該等小鼠。根據標準試驗計劃藉由劑量反應實驗確定HDAC抑制劑之最佳劑量。根據標準方法(例如Meyer等人，Int.J.Cancer 43：851－856(1989))每次注射後第二日計算腫瘤體積。以本發明之HDAC抑制劑進行處理相對於僅以媒劑(即無HDAC抑制劑)處理之對照組而言顯著減少腫瘤重量及體積。BCD1 in eight to ten week old female mice (Taconic Labs, Great Barrington, NY ) were injected subcutaneously flank region 2 x 10 6 pre-warp the modified HCT116 human colorectal cancer cells, SW48 colon cancer cells, and A549 lung cancer cells. The cells were pre-modified by at least three consecutive tumor transplants in the same strain of nude mice. Approximately 30 mg of the tumor fragment was subsequently excised and subcutaneously implanted into the left flank region of the mouse under Forene anesthesia (Abbott Labs, Geneva, Switzerland). When the average tumor volume reached 100 mm 3 , the mice were treated intraperitoneally by daily injection of a 10 mg/kg histone deacetylase inhibitor in DMSO. The optimal dose of HDAC inhibitor was determined by dose response experiments according to standard test protocols. Tumor volume was calculated on the second day after each injection according to standard methods (e.g., Meyer et al, Int. J. Cancer 43: 851-856 (1989)). Treatment with the HDAC inhibitors of the invention significantly reduced tumor weight and volume relative to control groups treated only with vehicle (i.e., no HDAC inhibitor).
本實例之目的係說明組合使用本發明之組蛋白去乙醯基酶抑制劑與組蛋白去乙醯基酶反義寡核苷酸增強對哺乳動物體內腫瘤生長之抑制作用的能力。較佳該反義寡核苷酸與該HDAC抑制劑抑制相同組蛋白去乙醯基酶之表現及活性。The purpose of this example is to demonstrate the ability to combine the histone deacetylase inhibitor of the present invention with a histone deacetylase antisense oligonucleotide to enhance the inhibition of tumor growth in a mammal. Preferably, the antisense oligonucleotide inhibits the expression and activity of the same histone deacetylase with the HDAC inhibitor.
以含有每公斤體重約0.1毫克至約30毫克組蛋白去乙醯基酶反義寡核苷酸之鹽水製劑每日處理承受經植入之HCT116腫瘤(平均體積100立方毫米)的小鼠。以含有每公斤體重約0.01毫克至約5毫克HDAC抑制劑之醫藥學上可接受之製劑每日處理第二組小鼠。Mice bearing the implanted HCT116 tumor (mean volume 100 mm3) were treated daily with a saline formulation containing from about 0.1 mg to about 30 mg of histone deacetylase antisense oligonucleotide per kg of body weight. The second group of mice is treated daily with a pharmaceutically acceptable formulation containing from about 0.01 mg to about 5 mg of HDAC inhibitor per kg of body weight.
一些小鼠既接受反義寡核苷酸又接受HDAC抑制劑。一組該等小鼠可經由尾部靜脈同時經靜脈接受反義寡核苷酸及HDAC抑制劑。另一組可經由尾部靜脈接受反義寡核苷酸，且皮下接受HDAC抑制劑。又一組可皮下接受反義寡核苷酸及HDAC抑制劑兩者。類似建立下述小鼠對照組：未接受處理(例如僅鹽水)，僅接受錯配反義寡核苷酸，接受不抑制組蛋白去乙醯基酶活性之對照化合物及接受錯配反義寡核苷酸與一對照化合物。Some mice receive both antisense oligonucleotides and HDAC inhibitors. A group of such mice can receive both antisense oligonucleotides and HDAC inhibitors via the tail vein via the vein. Another group can receive antisense oligonucleotides via the tail vein and receive HDAC inhibitors subcutaneously. Another group can receive both antisense oligonucleotides and HDAC inhibitors subcutaneously. Similar to the establishment of the following mouse control group: no treatment (eg saline only), only mismatched antisense oligonucleotides, control compounds that did not inhibit histone deacetylase activity, and mismatched antisense oligos Nucleotide and a control compound.
以卡尺量測腫瘤體積。相對於對照組而言，經以反義寡核苷酸加本發明之組蛋白去乙醯基酶蛋白抑制劑處理會顯著減少腫瘤重量及體積。The tumor volume was measured with a caliper. Treatment with antisense oligonucleotides plus the histone deacetylase inhibitor of the present invention significantly reduced tumor weight and volume relative to the control group.
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