TWI602818B - Fused heterocyclic compounds as protein kinase inhibitors - Google Patents
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本發明係關係一種稠合雜環化合物作為蛋白激酶抑制劑。 The present invention relates to a fused heterocyclic compound as a protein kinase inhibitor.
布魯頓酪氨酸激酶(Bruton’s tyrosine kinase,Btk)屬於Tec酪氨酸激酶家族(Vetrie et al.,Nature 361：226-233,1993；Bradshaw,Cell Signal. 22：1175-84,2010)。Btk主要表達在大多數造血細胞中，如B細胞，肥大細胞和巨噬細胞(Smith et al.,J.Immunol. 152：557-565,1994)，存在於骨髓，脾臟和淋巴結組織。Btk在B-細胞受體(B-cell receptor,BCR)和FcR信號通路中起著重要的作用，這些通路參與B-細胞發育，分化 (Khan,Immunol.Res. 23：147,2001)。Btk被上游Src-家族激酶啟動。一旦被啟動，Btk進一步磷酸化PLCγ，進而影響B-細胞的功能和存活(Humphries et al.,J.Biol.Chem. 279：37651,2004)。這些信號通路必須被精確地調節。人類表達Btk基因編碼的基因突變會導致遺傳性B-細胞特異性免疫缺陷疾病，被稱為X-連鎖無丙種球蛋白血症(X-linked agammaglobulinemia,XLA)(Conley et al.,Annu.Rev.Immunol. 27：199-227,2009)。BCR-介導信號的異常可能會導致B細胞活化的失調，導致許多自身免疫性疾病和炎症疾病。臨床前研究表明，Btk缺陷小鼠對膠原誘導性關節炎的發展有阻抗性。此外，臨床研究表明，Rituxan作為CD20抗體，用以減少成熟的B細胞，揭示了B-細胞在許多炎症疾病，如風濕性關節炎，全身性紅斑狼瘡，多發性硬化症中起著關鍵作用(Gurcan et al.,Int.Immunopharmacol. 9：10-25,2009)。因此，Btk抑制劑可用於治療自身免疫性疾病和/或炎症疾病。 Bruton's tyrosine kinase (Btk) belongs to the Tec tyrosine kinase family (Vetrie et al., Nature 361 : 226-233, 1993; Bradshaw, Cell Signal. 22 : 1175-84, 2010). Btk is mainly expressed in most hematopoietic cells, such as B cells, mast cells and macrophages (Smith et al., J. Immunol. 152: 557-565, 1994), present in bone marrow, spleen and lymph node tissues. Btk plays an important role in B-cell receptor (BCR) and FcR signaling pathways, which are involved in B-cell development and differentiation (Khan, Immunol. Res . 23 : 147, 2001). Btk is initiated by the upstream Src-family kinase. Once activated, Btk further phosphorylates PLC gamma, which in turn affects B-cell function and survival (Humphries et al., J. Biol. Chem. 279 :37651, 2004). These signal paths must be precisely adjusted. Mutations in human expression of the Btk gene cause a hereditary B-cell-specific immunodeficiency disease called X-linked agammaglobulinemia (XLA) (Conley et al., Annu . Rev .Immunol. 27 :199-227, 2009). Abnormalities in BCR-mediated signaling may lead to dysregulation of B cell activation leading to many autoimmune and inflammatory diseases. Preclinical studies have shown that Btk-deficient mice are resistant to the development of collagen-induced arthritis. In addition, clinical studies have shown that Rituxan acts as a CD20 antibody to reduce mature B cells, revealing that B-cells play a key role in many inflammatory diseases such as rheumatoid arthritis, systemic lupus erythematosus, and multiple sclerosis ( Gurcan et al., Int. Immunopharmacol . 9: 10-25, 2009). Thus, Btk inhibitors are useful in the treatment of autoimmune diseases and/or inflammatory diseases.
此外，Btk的異常活化在B-細胞淋巴瘤的發病機制中起著重要的作用，這意味著抑制Btk在血液惡性腫瘤的治療中是有很有用的(Davis et al.,Nature 463：88-92,2010)。初步臨床試驗結果顯示，布魯頓酪氨酸激酶(Bruton’s tyrosine kinase,Btk)抑制劑PCI-32765在治療幾種類型的B細胞淋巴瘤是有效的(例如，54th ASH annual meeting abstract,Dec.2012：686 The Bruton's Tyrosine Kinase(Btk)Inhibitor,Ibrutinib(PCI-32765),Has Preferential Activity in the ABC Subtype of Relapsed/Refractory De Novo Diffuse Large B-Cell Lymphoma (DLBCL)：Interim Results of a Multicenter,Open-Label,Phase2 Study)。由於Btk在調節多個信號轉導通路中起著核心作用，Btk抑制劑作為抗炎和/或抗癌藥物成為熱點。(Mohamed et al.,Immunol.Rev. 228：58-73,2009；Pan,Drug News perspect 21：357-362,2008；Rokosz et al.,Expert Opin.Ther.Targets 12：883-903,2008；Uckun et al.,Anti-cancer Agents Med. Chem. 7：624-632,2007；Lou et al.,J.Med.Chem. 55(10)：4539-4550,2012)。 In addition, abnormal activation of Btk plays an important role in the pathogenesis of B-cell lymphoma, which means that inhibition of Btk is useful in the treatment of hematological malignancies (Davis et al., Nature 463 :88- 92, 2010). Preliminary clinical trials have shown that Bruton's tyrosine kinase (Btk) inhibitor PCI-32765 is effective in the treatment of several types of B-cell lymphoma (eg, 54 th ASH annual meeting abstract, Dec. 2012: 686 The Bruton's Tyrosine Kinase (Btk) Inhibitor, Ibrutinib (PCI-32765), Has Preferential Activity in the ABC Subtype of Relapsed/Refractory De Novo Diffuse Large B-Cell Lymphoma (DLBCL): Interim Results of a Multicenter, Open- Label, Phase2 Study ). Since Btk plays a central role in regulating multiple signal transduction pathways, Btk inhibitors have become hot spots as anti-inflammatory and/or anti-cancer drugs. (Mohamed et al., Immunol. Rev. 228 : 58-73, 2009; Pan, Drug News perspect 21 : 357-362, 2008; Rokosz et al., Expert Opin . Ther . Targets 12 : 883-903, 2008; Uckun et al., Anti-cancer Agents Med. Chem. 7 : 624-632, 2007; Lou et al., J. Med. Chem. 55(10): 4539-4550, 2012).
許多正在開發的Btk小分子抑制劑用於炎症和癌症的治療。Ibrutinib(PCI-32765，參見：US7514444B2和相關檔，例如，US2012053189A1 WO2011153514和2011046964；US2010254905A1；WO2010009342，WO2008121742，WO2008054827；US20080139582；US20080076921；US7718662B1；WO2007087068；US20100035841)是首入臨床的一類Btk抑制劑，目前針對復發性或難治的套細胞淋巴瘤(mantle cell lymphoma,MCL)和慢性淋巴細胞白血病(chronic lymphocytic leukaemia,CLL)正在進行多個臨床試驗。另一個進入臨床試驗的Btk抑制劑是AVL-292，(參見，例如，US 20100249092；US20100029610；US2010016296；US20120077832，WO2011090760，WO2010028236，WO2009158571；WO2009051822，WO2010123870)。小野製藥 (Ono pharmaceuticals)和Mannkind公司分別在用自己的Btk小分子抑制劑進行著臨床試驗(參見，例如，ONO-4059，WO2011152351；WO2007136790A2)。 Many Btk small molecule inhibitors are being developed for the treatment of inflammation and cancer. Ibrutinib (PCI-32765, see: US7514444B2 and related files, for example, US2012053189A1 WO2011153514 and 2011046964; US2010254905A1; WO2010009342, WO2008121742, WO2008054827; US20080139582; US20080076921; US7718662B1; WO2007087068; US20100035841) are first-class clinical Btk inhibitors, currently targeted Recurrent or refractory mantle cell lymphoma (MCL) and chronic lymphocytic leukaemia (CLL) are undergoing several clinical trials. Another Btk inhibitor that enters clinical trials is AVL-292, (see, for example, US 20100249092; US20100029610; US2010016296; US20120077832, WO2011090760, WO2010028236, WO2009158571; WO2009051822, WO2010123870). Ono Pharmaceutical (Ono pharmaceuticals) and Mannkind, respectively, are conducting clinical trials with their own Btk small molecule inhibitors (see, for example, ONO-4059, WO2011152351; WO2007136790A2).
還有其他已知的Btk抑制劑。參見，例如，US2012/0232054(LOCUS製藥有限公司)，WO2010126960(LOCUS製藥有限公司)，WO 2011/162515(HANMI控股有限公司)，WO 2012135801(美國猶他州大學研究基金會)，Kim et al.,Bioorg.Med.Chem.Lett. 21：6258-6263,2011(輝瑞)，US8084620B2(BMS)，WO2002050071；WO2008116064；WO2010011837；WO 2011159857(BMS)，US2012058996A1；US2012082702A1 US20100160303(BMS)，US2012129852A1(BMS)，WO 2011019780(BMS)，WO 2011029043；WO 2011029046(Biogen Idec)，US7393848B2(CGI)，US20060178367；US20060183746(CGI)，EP2068849(CGI)，WO2005005429；WO2005014599；WO2005047290； WO2006053121；WO2008033834；WO2008033858；WO2006099075；WO2008033854；WO2008033857；WO2009039397(CGI)，WO2009137596；WO2010056875；WO2010068788；WO2010068806；WO2010068810(CGI，GENENTECH)，WO2011140488；WO2012030990；WO2012031004(GILEAD & GENENTECH)，US2012040961A1(Dana-Farber癌症研究所)，WO2005011597；WO2008045627；WO2008144253(IRM LLC)，WO2007140222；WO2013008095(NOVARTIS)，WO2012170976A2(Merck)，WO2012135944A1(PHARMASCIENCE)，US2010144705A1；US20120028981A1(PRINCIPIA BIOPHARMA)，WO2010065898A2；WO2012158795A1 WO2012158764A1 WO2012158810A1(PRINCIPIA BIOPHARMA)，US20090318448A1；US20100016301；US2009105209A1；US 20100222325；US20100004231(ROCHE)，WO2012156334A1； WO2012020008；WO2010122038；WO2010006970；WO2010006947；WO2010000633；WO2009077334；WO2009098144(ROCHE)，WO2006065946；WO2007027594；WO2007027729(VERTEX)。 There are other known Btk inhibitors. See, for example, US 2012/0232054 (LOCUS Pharmaceutical Co., Ltd.), WO2010126960 (LOCUS Pharmaceutical Co., Ltd.), WO 2011/162515 (HANMI Holdings Ltd.), WO 2012135801 (University of Utah Research Foundation), Kim et al., Bioorg. Med. Chem. Lett. 21 : 6258-6263, 2011 (Pfizer), US8084620B2 (BMS), WO2002050071; WO2008116064; WO2010011837; WO2011159857(BMS), US2012058996A1; US2012082702A1 US20100160303 (BMS), US2012129852A1 (BMS), WO 2011019780 (BMS), WO 2011029043; WO 2011029046 (Biogen Idec), US7393848B2 (CGI), US20060178367; US20060183746 (CGI), EP2068849 (CGI), WO2005005429; WO2005014599; WO2005047290; WO2006053121; WO2008033834; WO2008033858; WO2006099075; WO2008033854; WO2008033857; WO2009039397 (CGI), WO2009137596; WO2010056875; WO2010068788; WO2010068806; WO2010068810(CGI, GENENTECH), WO2011140488; WO2012030990; WO2012031004(GILEAD & GENENTECH), US2012040961A1 (Dana-Farber Cancer Institute), WO2005011597; WO2008045627; WO2008144253(IRM LLC) , WO2007140222; WO2013008095 ( NO 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 ; WO2010006970; WO2010006947; WO2010000633; WO2009077334; WO2009098144(ROCHE), WO2006065946; WO2007027594; WO2007027729 (VERTEX).
WO2007/026720 A1公開了稠合環吡唑化合物，由式(A)表示。其中，n表示2或3；A代表式：-O-或類似基團；B代表一個C1-10亞烷基或類似基團；C代表單鍵或式：-O-；R1-代表氫原子，吡咯烷基或類似基團；R4，R5和R6獨立地表示一個氫原子，一個鹵原子或類似基團；D1=D2代表式：-CH=CH-或類似基團；E代表式：-O-或-NH-或類似基團；G代表C1-10亞烷基或類似基團；R7代表一個氫原子，一個苯基或類似基團。這類化合物被用作Lck激酶抑制劑：
本發明提供了方法和組合物，用於抑制Btk和治療由不良的Btk活動引起的疾病(與Btk相關的疾病)。 The present invention provides methods and compositions for inhibiting Btk and treating diseases caused by undesirable Btk activity (Btk-associated diseases).
在實施例中，本發明提供了Btk抑制劑或式中的化合物：
及其立體異構體，和它們在藥學上可接受的鹽，其中：A是一個5-或6-元芳環，包括0-3個雜原子N，S或O；每個W是-CH2-或者-C(O)-；L是一個鍵，CH2，NR12，O，或者S；S/D是一個單鍵或者雙鍵；當是雙鍵時，R5和R7不存在；
m表示0或1-4的整數；n表示0或1-4的整數，其中，當n大於1時，每個R2可以不同；p是0，或1-2的整數，其中，當p為0時，m是非零的，並且當p大於1時，每個R6和每個R7可能是不同的；R1，R4，R5，R6和R7，分別選自氫，鹵素，雜烷基，烷基，烯基，環烷基，芳基，飽和或者不飽和的雜環基，雜芳基，炔基，-CN,-NR13R14,-OR13,-COR13,-CO2R13,-CONR13R14,-C(=NR13)NR14R15,-NR13COR14,-NR13CONR14R15,-NR13CO2R14,-SO2R13,-NR13SO2NR14R15或者-NR13SO2R14，其中，該烷基，烯基，炔基，環烷基，雜芳基，芳基和飽和或者不飽和的雜環基可以視需要被至少一個取代基R16任意取代，其中(R4和R5)，或(R4和R6)，或(R6和R7)，或(當p為2時，R6和R6)，連同它們所連接的原子，可以形成一個環，該環選自視需要至少被一個取代基R16任意取代的環烷基，飽和或者不飽和的雜環，芳基和雜芳環；R2選自鹵素，烷基，-S-烷基，-CN,-NR13R14,-OR13,-COR13,-CO2R13,-CONR13R14,-C(=NR13)NR14R15,-NR13COR14,-NR13CONR14R15,-NR13CO2R14,-SO2R13,-NR13SO2NR14R15或者-NR13SO2R14；R12選自氫和低級烷基；
R13，R14和R15，分別選自氫，雜烷基，烷基，烯基，炔基，環烷基，飽和或者不飽和的雜環基，芳基或者雜芳基；其中(R13和R14)，和/或(R14和R15)連同它們所連接的原子，每一個可以形成一個環，該環選自視需要至少被一個取代基R16任意取代的環烷基，飽和或者不飽和的雜環，芳基和雜芳環；R16選自鹵素，取代或未取代的烷基取代或未取代的烯基，取代或未取代的炔基，取代或未取代的環烷基，取代或未取代的芳基，取代或未取代的雜芳基，取代或未取代的飽和或者不飽和的雜環基，含氧的，-CN,-OR’,-NR’R”,-COR’,-CO2R’,-CONR’R”,-C(=NR’)NR”R''',-NR’COR”,-NR’CONR’R”,-NR’CO2R”,-SO2R’,-SO2aryl,-NR’SO2NR”R'''或者NR’SO2R”，其中，R’,R”和R'''各自獨立地選自氫，鹵素，取代或未取代的烷基，取代或未取代的烯基，取代或未取代的炔基，取代或未取代的環烷基，取代或未取代的芳基，取代或未取代的雜芳基，取代或未取代的雜環基，其中(R’和R”)，和/或者(R”和R''')連同它們所連接的原子，可以形成一個環，該環選自環烷基，飽和或者不飽和的雜環，芳基和雜芳環；在具體的實施例中：(a)S/D是一個雙鍵時，R5和R7不存在；(b)R1是氫，鹵素，烷氧基，雜烷基，烷基，烯基，環烷基，芳基，飽和或不飽和的雜環基，雜芳基，其中，烷基，烯
基，炔基，環烷基，雜芳基，芳基，飽和或不飽和的雜環基可以視需要被至少一個取代基R16任意取代；(c)p是1以及m是0，1或2，優選0或者1；(d)A是苯基；(e)每個R2獨立地選自鹵素，低級烷基，或低級烷氧基；(f)R4和R6，與它們所連接的原子，形成一個環，該環選自視需要被至少一個取代基R16任意取代的環烷基，飽和或不飽和的雜環，芳基，和雜芳基；(g)R4和R6，與它們所連接的原子，形成一個式中的環：
(h)S/D是一個單鍵。 (h) S/D is a single button.
(i)p是0，且R6和R7不存在。 (i) p is 0, and R 6 and R 7 are absent.
本發明包括所列舉的具體實施例的所有組合，例如，上面的(a)-(i)，就像每個組合分別進行了逐個列舉。 The present invention includes all combinations of the specific embodiments enumerated, for example, (a)-(i) above, as each combination is recited separately.
具體的實施例的典型組合： (i)S/D是一個雙鍵，且R5和R7不存在；R1是氫，鹵素，烷氧基，雜烷基，烷基，烯基，環烷基，芳基，飽和或不飽和的雜環基，雜芳基，其中，烷基，烯基，炔基，環烷基，雜芳基，芳基，飽和或不飽和的雜環基可以視需要被至少一個取代基R16任意取代；且R16是鹵素，低級烷基，或低級烷氧基；(ii)S/D是一個雙鍵，且R5和R7不存在；p是1且m是0，1(或2)；(iii)S/D是一個雙鍵，且R5和R7不存在；p是1以及m是0，1(或2)；A是苯基；以及每個R2獨立地選自鹵素，低級烷基，或低級烷氧基(參見，式II)；(iv)S/D是一個雙鍵，且R5和R7不存在；以及R4和R6，與它們所連接的原子，形成一個環，該環選自視需要被至少一個取代基R16任意取代的環烷基，飽和或不飽和的雜環，芳基，和雜芳基。 A typical combination of specific examples: ( i ) S/D is a double bond and R 5 and R 7 are absent; R 1 is hydrogen, halogen, alkoxy, heteroalkyl, alkyl, alkenyl, ring Alkyl, aryl, saturated or unsaturated heterocyclic group, heteroaryl, wherein alkyl, alkenyl, alkynyl, cycloalkyl, heteroaryl, aryl, saturated or unsaturated heterocyclic group Optionally substituted with at least one substituent R 16 ; and R 16 is halo, lower alkyl, or lower alkoxy; ( ii ) S/D is a double bond and R 5 and R 7 are absent; p is 1 and m is 0,1 (or 2); ( iii ) S/D is a double bond, and R 5 and R 7 are absent; p is 1 and m is 0,1 (or 2); A is phenyl And each R 2 is independently selected from halogen, lower alkyl, or lower alkoxy (see, formula II ); ( iv ) S/D is a double bond, and R 5 and R 7 are absent; 4 and R 6, with the atoms to which they are attached, form a ring, the ring is selected from optionally substituted with at least one R 16 group optionally substituted cycloalkyl, saturated or unsaturated heterocycle, aryl, and heteroaryl base.
(v)S/D是一個雙鍵，且R5和R7不存在；R4和R6，與它們所連接的原子，形成一個環，該環選自視需要被至少一個取代基R16任意取代的環烷基，飽和或不飽和的雜環，芳基，和雜芳基；A是苯基；以及每個R2獨立地選自鹵素，低級烷基，或低級烷氧基。 ( v ) S/D is a double bond, and R 5 and R 7 are absent; R 4 and R 6 , with the atom to which they are attached, form a ring selected from at least one substituent R 16 as desired. Optionally substituted cycloalkyl, saturated or unsaturated heterocyclic, aryl, and heteroaryl; A is phenyl; and each R 2 is independently selected from halo, lower alkyl, or lower alkoxy.
(vi)S/D是一個雙鍵，且R5和R7不存在；R4和R6，與它們所連接的原子，形成一個環，該環選自視需要被至少一個取代基R16任意取代的環烷基，飽和或不飽和的雜環，芳基，和雜芳
基。p是1以及m是0，1(或2)；A是苯基；以及每個R2獨立地選自鹵素，低級烷基，或低級烷氧基；以及R4-R6環是式：
本發明還提供了實施例，或者表I，II或III(下面)中的化合物，及其立體異構體，和它們在藥學上可接受的鹽。 The invention also provides examples, or compounds of Tables I, II or III (below), and stereoisomers thereof, and pharmaceutically acceptable salts thereof.
本發明還提供具有抑制Btk活性的化合物，化合物在BTK激酶檢測中具有10uM或以下的IC50。 The present invention also provides a compound having an activity of inhibiting Btk, which has an IC50 of 10 uM or less in BTK kinase assay.
本發明還提供了藥物組合物，該組合物包括治療有效量的單位劑量的本發明中的化合物和一種或多種藥學上可接受的載體。 The invention also provides a pharmaceutical composition comprising a therapeutically effective amount of a unit dose of a compound of the invention and one or more pharmaceutically acceptable carriers.
本發明還提供了聯合給藥，包括一個治療有效量的本發明中的化合物和一個不同的能夠有效治療自身免疫性疾病和/或炎症性疾病的試劑。 The invention also provides co-administration comprising a therapeutically effective amount of a compound of the invention and a different agent effective to treat an autoimmune disease and/or an inflammatory disease.
本發明還提供方法，該方法能夠治療Btk相關的疾病，或由於Btk異常活化引起的疾病，特別是過敏性疾病，自身免疫性疾病(例如，類風濕關節炎)，炎性疾病，或癌症(例如：B-細胞增殖性疾病，如慢性淋巴細胞性淋巴瘤，非霍奇金淋巴瘤，瀰漫性大B-細胞淋巴瘤，套細胞淋巴瘤，濾泡性淋巴瘤或慢性淋巴細胞白血病)。該方法一般包括向需要的哺乳動物給藥有效量的一種本發 明中的化合物，其N-氧化物或它們的前藥，可以任選地用於檢測所述疾病或者症狀的改善情況，或者抑制Btk。 The present invention also provides a method capable of treating a Btk-related disease, or a disease caused by abnormal activation of Btk, particularly an allergic disease, an autoimmune disease (for example, rheumatoid arthritis), an inflammatory disease, or cancer ( For example: B-cell proliferative diseases such as chronic lymphocytic lymphoma, non-Hodgkin's lymphoma, diffuse large B-cell lymphoma, mantle cell lymphoma, follicular lymphoma or chronic lymphocytic leukemia). The method generally comprises administering to a mammal in need thereof an effective amount of a hair The compounds of the formula, their N-oxides or their prodrugs, can optionally be used to detect an improvement in the disease or condition, or to inhibit Btk.
本發明還提供了藥物組合物，該組合物包括單位劑量的，一定給藥形式的本發明中的化合物，和誘導細胞自噬的方法，該方法包括向需要的人給藥有效量的本發明中的一種化合物或組合物。 The invention also provides a pharmaceutical composition comprising a unit dosage, a compound of the invention in a certain dosage form, and a method of inducing autophagy in a method comprising administering to a human in need thereof an effective amount of the invention a compound or composition.
本發明也提供了本發明中的化合物用作藥物，以及使用本發明中的化合物製造一種治療Btk相關疾病的藥物。 The present invention also provides a compound of the present invention for use as a medicament, and a medicament for treating a Btk-related disease using the compound of the present invention.
本發明公開的化合物，能夠抑制酪氨酸激酶，例如Btk,Blk,Bmx,EGFR,ERBB2,ERBB4,Itk,Jak3,Tec和Txk激酶。 The compounds disclosed herein are capable of inhibiting tyrosine kinases such as Btk, Blk, Bmx, EGFR, ERBB2, ERBB4, Itk, Jak3, Tec and Txk kinases.
在本發明中，除非上下文中另有所指，下面所要用到的單詞，短語和符號，要表達的含義有如下規定。以下縮寫和術語的含義貫穿整文：術語“烷基”是指烴基，所述烴基選自飽和的直鏈的和帶支鏈的烴基，所述烴基包括1到18，或者1到12，或者1到6個碳原子。烷基的實例包括甲基，乙基，1-丙基或正丙基("n-Pr")，2-丙基或異丙基("i-Pr")，1-丁基或正-丁基("n-Bu")，2-甲基-1-丙基或異丁基("i-Bu")，1-甲丙基或仲丁基("s-Bu")，以及1,1-二甲基乙基或叔丁基("t-Bu")。其他的烷基的例子包括1-戊基， 2-戊基，3-戊基，2-甲基-2-丁基，3-甲基-2-丁基，3-甲基-1-丁基，2-甲基-1-丁基，1-己基，2-己基，3-己基，2-甲基-2-戊基，3-甲基-2-戊基，4-甲基-2-戊基，3-甲基-3-戊基，2-甲基-3-戊基，2,3-二甲基-2-丁基和3,3-二甲基-2-丁基等基團。 In the present invention, unless otherwise indicated in the context, the meanings of the words, phrases and symbols to be used below are as follows. The following abbreviations and terms have the meaning throughout the text: the term "alkyl" refers to a hydrocarbyl group selected from saturated straight-chain and branched hydrocarbyl groups, including from 1 to 18, or from 1 to 12, or 1 to 6 carbon atoms. Examples of alkyl groups include methyl, ethyl, 1-propyl or n-propyl ("n-Pr"), 2-propyl or isopropyl ("i-Pr"), 1-butyl or n- Butyl ("n-Bu"), 2-methyl-1-propyl or isobutyl ("i-Bu"), 1-methylpropyl or sec-butyl ("s-Bu"), and 1 , 1-dimethylethyl or tert-butyl ("t-Bu"). Examples of other alkyl groups include 1-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl a group such as 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl and 3,3-dimethyl-2-butyl.
低級烷基是指1-8，優選1-6，更優選為1-4個碳原子；低級烯基或炔基是指2-8，2-6或2-4個碳原子。 Lower alkyl means 1-8, preferably 1-6, more preferably 1-4 carbon atoms; lower alkenyl or alkynyl means 2-8, 2-6 or 2-4 carbon atoms.
這裡的術語“烯基”指的是選自直鏈的和帶支鏈的烴基，所述烴基包括至少一個C=C雙鍵和2到18，或者2到12，或者2到6個碳原子。所述烯基的例子可以選自於亞乙基或乙烯基，1-丙烯基，2-丙烯基，2-甲基-1-丙烯基，1-丁烯基，2-丁烯基，3-丁烯基，1,3-丁二烯基，2-甲基-1,3-丁二烯基，1-正丁烯基，2-正丁烯基，3-正丁烯基，4-正丁烯基，和1,3-正己二烯基基團。 The term "alkenyl" as used herein refers to a hydrocarbon group selected from the group consisting of a straight chain and a branched chain, the hydrocarbon group comprising at least one C=C double bond and 2 to 18, or 2 to 12, or 2 to 6 carbon atoms. . Examples of the alkenyl group may be selected from ethylene or vinyl, 1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, 3 -butenyl, 1,3-butadienyl, 2-methyl-1,3-butadienyl, 1-n-butenyl, 2-n-butenyl, 3-n-butenyl, 4 - n-butenyl, and 1,3-n-hexadienyl groups.
這裡的術語“炔基”指的是選自直鏈的和帶支鏈的烴基，所述烴基包括至少一個C≡C三鍵和2到18，或者2到12，或者2到6個碳原子。所述炔基的例子包括乙炔基，1-丙炔基，2-丙炔基(炔丙基)，1-丁炔基，2-丁炔基，和3-丁炔基基團。 The term "alkynyl" as used herein refers to a hydrocarbon group selected from the group consisting of a straight chain and a branched chain, the hydrocarbon group comprising at least one C≡C triple bond and 2 to 18, or 2 to 12, or 2 to 6 carbon atoms. . Examples of the alkynyl group include an ethynyl group, a 1-propynyl group, a 2-propynyl (propargyl group), a 1-butynyl group, a 2-butynyl group, and a 3-butynyl group.
這裡的術語“環烷基”指的是選自飽和和部分不飽和的環烴基，所述環烴基包括單環的和多環(例如，雙環的和三環的)基團。例如，所述環烷基可以包括3到12，或者3到8，或者3到6個碳原子。進一步例如，環烷基可以是3到12，或者3到8，或者3到6個碳原子的單環基團。單環環烷烴基的例子包括環丙基，環丁基，環戊基，1-環戊烷-1-烯基，1-環戊烷-2-烯基，1-環戊烷-3- 烯基，環己基，1-環己基-1-烯基，1-環己基-2-烯基，1-環己基-3-烯基，環己二烯基，環庚基，環辛基，環壬基，環癸基，環十一烷基，和環十二烷基。雙環環烷基的例子包括由7到12個環原子排列組成的雙環基或橋雙環基，所述雙環選自[4,4],[4,5],[5,5],[5,6]和[6,6]環系，所述橋雙環選自雙環[2.2.1]庚烷，雙環[2.2.2]辛烷，和雙環[3.2.2]壬烷。所述的環可以是飽和的或具有至少一個雙鍵(比如，部分不飽和)，但不是完全共軛的，且不是芳香族的，如本文所定義的芳香族。 The term "cycloalkyl" as used herein refers to a cyclic hydrocarbon group selected from the group consisting of saturated and partially unsaturated, including cyclocyclic and polycyclic (eg, bicyclic and tricyclic) groups. For example, the cycloalkyl group can include from 3 to 12, or from 3 to 8, or from 3 to 6 carbon atoms. Further, for example, the cycloalkyl group may be a monocyclic group of 3 to 12, or 3 to 8, or 3 to 6 carbon atoms. Examples of monocyclic cycloalkane groups include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopentan-1-enyl, 1-cyclopentan-2-enyl, 1-cyclopentane-3- Alkenyl, cyclohexyl, 1-cyclohexyl-1-enyl, 1-cyclohexyl-2-alkenyl, 1-cyclohexyl-3-alkenyl, cyclohexadienyl, cycloheptyl, cyclooctyl, Cyclodecyl, cyclodecyl, cyclodecyl, and cyclododecyl. Examples of the bicyclic cycloalkyl group include a bicyclic group or a bridged bicyclic group consisting of 7 to 12 ring atoms selected from [4, 4], [4, 5], [5, 5], [5, 6] and [6,6] ring system selected from bicyclo [2.2.1] heptane, bicyclo [2.2.2] octane, and bicyclo [3.2.2] decane. The ring may be saturated or have at least one double bond (eg, partially unsaturated), but is not fully conjugated and is not aromatic, as defined herein.
這裡的術語“芳香基”指一個基團選自：5-元和6-元的碳環芳香環，例如，苯基；雙環體系如7-元到12-元的雙環系，其中至少有一個環是碳環和芳香環，如所述雙環體系選自例如萘，茚和1,2,3,4-四氫喹啉；和三環體系如10到15元三環體系，其中至少有一個環是碳環和芳香環，如芴。 The term "aryl" as used herein refers to a group selected from the group consisting of 5-membered and 6-membered carbocyclic aromatic rings, for example, phenyl; bicyclic systems such as 7- to 12-membered bicyclic systems, at least one of which The ring is a carbocyclic ring and an aromatic ring, such as the bicyclic ring system selected from, for example, naphthalene, anthracene and 1,2,3,4-tetrahydroquinoline; and a tricyclic system such as a 10 to 15 membered tricyclic system, at least one of which The ring is a carbocyclic ring and an aromatic ring such as hydrazine.
例如，所述芳基是選自於將5-元和6-元碳環芳香環並到5-元到7-元環烷基或雜環上形成的芳基，所述5-元到7-元環烷基或雜環包含至少一個選自N,O和S的雜原子，如果所述碳芳香環並一個雜環那麼連接點在碳芳香環上，如果所述的碳芳香環與環烷基並環那麼連接點可以在碳芳香環或環烷基上。形成於取代的苯基衍生物而且在環原子中具有自由價的二價基被稱為取代的亞苯基自由基。單價多環烴基命名以“-基”結尾，由單價多環烴基從具有自由價的碳原子上移去一個氫原子得到二價基，命名是在相應的單價基的名稱中加入“-亞基"，如將有兩個連接點的萘基稱作亞二氫萘 基。然而，無論如何芳基都不包括或與雜芳基重疊，下面將分別定義。因此，如果一個或多個碳芳香環與雜環芳香環稠合，所得到的環體系為本發明中定義的雜芳基，而非芳基。 For example, the aryl group is an aryl group selected from the group consisting of a 5-membered and a 6-membered carbocyclic aromatic ring and bonded to a 5-membered to 7-membered cycloalkyl or heterocyclic ring, said 5-member to 7 a cycloalkylene or heterocyclic ring containing at least one hetero atom selected from N, O and S, if the carbon aromatic ring and a heterocyclic ring are attached to a carbon aromatic ring, if said carbon aromatic ring and ring The alkyl-cyclo ring can be attached to a carbon aromatic ring or a cycloalkyl group. A divalent group formed on a substituted phenyl derivative and having a free valence in a ring atom is referred to as a substituted phenylene radical. The monovalent polycyclic hydrocarbon group is terminated by a "- group", and a monovalent polycyclic hydrocarbon group is removed from a carbon atom having a free valence to obtain a divalent group, and a "subunit" is added to the name of the corresponding monovalent group. ", such as the naphthyl group with two points of attachment, called dihydronaphthalene base. However, in any case, the aryl group does not include or overlap with the heteroaryl group, which will be defined separately below. Thus, if one or more carbon aromatic rings are fused to a heterocyclic aromatic ring, the resulting ring system is a heteroaryl group as defined in the invention, rather than an aryl group.
這裡的術語“鹵素”或“鹵”指的是F,Cl,Br或I。 The term "halogen" or "halo" as used herein refers to F, Cl, Br or I.
術語“雜烷基”是指包括至少一個雜原子的烷基。 The term "heteroalkyl" refers to an alkyl group comprising at least one hetero atom.
這裡的術語“雜芳基”選自於： 5到7元芳香的單環，包含1,2,3或者4個雜原子，該雜原子選自N,O和S，其餘的環原子為碳原子；8元到12元雙環，包括1,2,3或者4個雜原子，該雜原子選自N,O和S，其餘的環原子為碳原子，並且其中至少一個環是芳香族的，並且芳香環上至少有一個雜原子；11到14元三環，包括1,2,3或者4個雜原子，該雜原子選自N,O，和S，其餘的環原子是碳並且其中至少一個環是芳香族的，並且芳香環上至少有一個雜原子。 The term "heteroaryl" as used herein is selected from: A 5- to 7-membered aromatic monocyclic ring containing 1, 2, 3 or 4 heteroatoms selected from N, O and S, the remaining ring atoms being carbon atoms; 8 to 12 membered bicyclic rings, including 1, 2, 3 or 4 heteroatoms selected from N, O and S, the remaining ring atoms being carbon atoms, and at least one of which is aromatic and having at least one heteroatom on the aromatic ring; a 14-membered tricyclic ring comprising 1, 2, 3 or 4 heteroatoms selected from N, O, and S, the remaining ring atoms being carbon and at least one of which is aromatic and at least on the aromatic ring There is a hetero atom.
例如，雜芳基包括5到7元雜芳環並上5到7元環烷基環。對於這樣的並環，雙雜芳環體系中只是其中的一個環包含至少一個雜原子，連接點可以在雜芳環或環烷基環。 For example, a heteroaryl group includes a 5 to 7 membered heteroaryl ring and a 5 to 7 membered cycloalkyl ring. For such a bicyclic ring, only one of the rings in the biheteroaromatic ring system contains at least one hetero atom, and the point of attachment may be in a heteroaryl ring or a cycloalkyl ring.
當雜芳基上的S和O原子的總數超過1，這些雜原子就不會相鄰。在一些實施例中，雜芳基上的S和O的原子總數不超過2；在另一些實施例中，雜芳環上S和O的原子總數不超過1。 When the total number of S and O atoms on the heteroaryl group exceeds 1, these heteroatoms are not adjacent. In some embodiments, the total number of atoms of S and O on the heteroaryl group does not exceed 2; in other embodiments, the total number of atoms of S and O on the heteroaryl ring does not exceed one.
雜芳基的例子包括，但不局限於(從優先指定的連接位置編碼1)吡啶基(例如2-吡啶基，3-吡啶基，4-吡啶基)，噌啉 基，吡嗪基，2,4-嘧啶基，3,5-嘧啶基，2,4-咪唑基，咪唑並吡啶基，異惡唑基，惡唑基，噻唑基，異噻唑基，噻二唑，四唑基，噻吩基，三嗪基，苯並噻吩基，呋喃基，苯並呋喃基，苯並咪唑基，吲哚基，異吲哚基，二氫吲哚基，酞嗪基，吡嗪基，噠嗪基，吡咯基，***基，喹啉基，異喹啉基，吡唑基，吡咯並吡啶基(比如1H-吡咯並[2,3-b]吡啶-5-基)，吡唑並吡啶基(比如1H-吡唑並[3,4-b]吡啶-5-基)，苯並惡唑基(比如苯並[d]惡唑-6-基)，蝶啶基，嘌呤基，1-氧雜-2,3-二唑基，1-氧雜-2,4-二唑基，1-氧雜-2,5-二唑基，1-氧雜-3,4-二唑基，1-硫代-2,3-二唑基，1-硫代-2,4-二唑基，1-硫代-2,5-二唑基，1-硫代-3,4-二唑基，呋吖基，苯並呋吖基，苯並噻吩基，苯並噻唑基，苯並惡唑基，喹唑啉基，喹惡啉基，萘啶基，呋喃並吡啶基，苯並噻唑基(比如苯並[d]噻唑-6-基)，吲哚基(比如1H-吲唑-5-基)和5,6,7,8-四氫異喹啉。 Examples of heteroaryl groups include, but are not limited to, (encoded from a preferentially specified position of 1) pyridyl (e.g., 2-pyridyl, 3-pyridyl, 4-pyridyl), porphyrin , pyrazinyl, 2,4-pyrimidinyl, 3,5-pyrimidinyl, 2,4-imidazolyl, imidazopyridyl, isoxazolyl, oxazolyl, thiazolyl, isothiazolyl, thia Azole, tetrazolyl, thienyl, triazinyl, benzothienyl, furyl, benzofuranyl, benzimidazolyl, fluorenyl, isodecyl, indanyl, pyridazinyl, Pyrazinyl, pyridazinyl, pyrrolyl, triazolyl, quinolyl, isoquinolinyl, pyrazolyl, pyrrolidopyridyl (such as 1H-pyrrolo[2,3-b]pyridin-5-yl , pyrazolopyridinyl (such as 1H-pyrazolo[3,4-b]pyridin-5-yl), benzoxazolyl (such as benzo[d]oxazole-6-yl), pteridine Base, fluorenyl, 1-oxa-2,3-oxadiazolyl, 1-oxa-2,4-oxadiazole, 1-oxa-2,5-oxadiazolyl, 1-oxa-3 ,4-oxazolyl, 1-thioxo-2,3-diazolyl, 1-thio-2,4-oxadiazole, 1-thio-2,5-oxadiazolyl, 1-thio -3,4-oxadiyl, furazyl, benzofurazinyl, benzothienyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridyl, furan Pyridyl, benzothiazolyl (such as benzo[d]thiazole-6-yl), fluorenyl (such as 1H-carbazol-5-yl) and 5,6,7,8-tetrahydroisoquinoline.
這裡的術語“雜環的”或“雜環”或“雜環基”指的是選自4到12元的單環，雙環，三環的一類環，所述雜環是飽和和部分不飽和環，包括除選自N,O和S中的1,2,3或者4個雜原子外，至少一個碳原子。這裡的“雜環”也指一類5-到7-元雜環，所述雜環由至少包括一個選自N,O和S的雜原子，且與5-，6-，和/或7-元的環烷基環，碳環芳香環或雜芳環並環，當所述雜環與一個碳環芳香環或一個雜芳環並環時連接點在雜環上，而且當所述雜環與環烷基並環時連接點可以是在環烷基或雜環上。 The term "heterocyclic" or "heterocyclic" or "heterocyclyl" as used herein, refers to a monocyclic, bicyclic, tricyclic ring of one to four members selected from the group consisting of saturated and partially unsaturated. The ring includes at least one carbon atom in addition to 1, 2, 3 or 4 heteroatoms selected from N, O and S. "Heterocycle" as used herein also refers to a class of 5- to 7-membered heterocyclic rings consisting of at least one hetero atom selected from N, O and S, and 5-, 6-, and/or 7- a cycloalkyl ring, a carbocyclic aromatic ring or a heteroaryl ring-and-ring, when the heterocyclic ring is ring-bonded to a carbocyclic aromatic ring or a heteroaromatic ring, the point of attachment is on the heterocyclic ring, and when the heterocyclic ring The point of attachment to the cycloalkyl ring may be on a cycloalkyl or heterocycle.
這裡的“雜環”也指一類脂肪族的螺環，所述螺環包括至少一個選自N,O，和S的雜原子，連接點在所述雜環上。上述的這些環可能是飽和的或含有至少一個雙鍵(也就是部分不飽和)。上述雜環可能被氧取代。連接點可能是雜環上的碳原子或雜原子。雜環不是這裡所定義的雜芳環。 "Heterocycle" as used herein also refers to a class of aliphatic spiro rings comprising at least one heteroatom selected from the group consisting of N, O, and S, to which the point of attachment is attached. These rings may be saturated or contain at least one double bond (ie, partially unsaturated). The above heterocyclic ring may be substituted by oxygen. The point of attachment may be a carbon atom or a hetero atom on the heterocycle. A heterocyclic ring is not a heteroaryl ring as defined herein.
雜環的例子包括，但不局限於(從優先指定的連接位置編碼1)1-吡咯烷基，2-吡咯烷基，2,4-咪唑烷基，2,3-吡唑烷基，1-哌啶基，2-哌啶基，3-哌啶基，4-哌啶基，2,5-哌嗪基，吡喃基，2-嗎啉基，3-嗎啉基，環氧乙烷基，氮雜環丙烯基，環硫乙烷基，氮雜環丁基，氧雜環丁烷基，硫雜環丁基，1,2-二硫雜環丁基，1,3-二硫雜環丁基，二氫吡啶，四氫吡啶，硫代嗎啉，氧硫雜環己烷基，哌嗪基，高哌嗪基，高哌啶基，氮雜環庚烷基，氧雜環庚烷基，硫雜環庚烷基，1,4-氧硫雜環己烷基，1,4-二氧雜環庚烷基，1,4-氧硫雜環庚烷基，1,4-氮氧雜環庚烷基，1,4-二硫雜環庚烷基，1,4-氮硫雜環庚烷基和1,4-二氮雜環庚烷，1-1,4-二噻烷基，1,4-氮硫雜環己烷基，氧氮雜卓，二氮雜卓，硫氮雜卓，二氫噻吩基，二氫吡喃基，二氫呋喃基，四氫呋喃基，四氫噻吩基，四氫吡喃基，四氫噻喃基，1-吡咯啉基，2-吡咯啉基，3-吡咯啉基，吲哚啉基，2H-吡喃基，4H-吡喃基，1,4-二氧雜環己烷基，1,3-二氧環戊基，吡唑啉基，吡唑烷基，二噻烷基，二噻環戊基，吡唑烷基，咪唑啉基(pyrazolidinylimidazolinyl)，嘧啶酮基，1,1-二氧代-硫代嗎啉基，3-氮雜雙環[3.1.0]己基，3-氮雜雙環[4.1.0]庚烷基，氮雜雙 環[2.2.2]己基。取代的雜環基還包括一個或多個氧代基團取代的環體系，比如N-氧化哌啶基，N-氧化嗎啉基，1-氧代-1-硫代嗎啉基和1,1-二氧代-1-硫代嗎啉基。 Examples of heterocycles include, but are not limited to, (encoded from a preferred position of the linkage 1) 1-pyrrolidinyl, 2-pyrrolidinyl, 2,4-imidazolidinyl, 2,3-pyrazolyl, 1 -piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl, 2,5-piperazinyl, pyranyl, 2-morpholinyl, 3-morpholinyl, epoxy Alkyl, azacyclopropenyl, cyclohexylethane, azetidinyl, oxetanyl, thioheterobutyl, 1,2-dithiot-butyl, 1,3-di Thiopyletyl, dihydropyridine, tetrahydropyridine, thiomorpholine, oxathiane, piperazinyl, homopiperazinyl, homopiperidinyl, azepanyl, oxa Cycloheptyl, thiaheptanyl, 1,4-oxathiane, 1,4-dioxanyl, 1,4-oxathiamethylene, 1, 4-azacycloheptane, 1,4-dithiaheptanyl, 1,4-azetidinyl and 1,4-diazepane, 1-1, 4 -dithiaalkyl, 1,4-azetidinyl, oxazepine, diazepine, thiazepine, dihydrothienyl, dihydropyranyl, dihydrofuranyl, tetrahydrofuran Base, tetrahydrothiophenyl, tetrahydropyranyl, tetra Thianyl, 1-pyrroline, 2-pyrolinyl, 3-pyrrolyl, porphyrin, 2H-pyranyl, 4H-pyranyl, 1,4-dioxanyl , 1,3-dioxocyclopentyl, pyrazolinyl, pyrazolidinyl, dithiaalkyl, dithiacyclopentyl, pyrazolidinyl, pyrazolidinylimidazolinyl, pyrimidinone, 1, 1-dioxo-thiomorpholinyl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptanyl, aza double Ring [2.2.2] hexyl. Substituted heterocyclic groups also include one or more oxo group substituted ring systems such as N-oxypiperidinyl, N-oxymorpholinyl, 1-oxo-1-thiomorpholinyl and 1, 1-dioxo-1-thiomorpholinyl.
取代基選自：鹵素，-R',-OR',=O,=NR',=N-OR',-NR'R",-SR',-SiR'R"R'",-OC(O)R',-C(O)R',-CO2R',-CONR'R",-OC(O)NR'R",-NR"C(O)R',-NR'-C(O)NR"R'",-NR'-SO2NR'",-NR"CO2R',-NH-C(NH2)=NH,-NR'C(NH2)=NH,-NH-C(NH2)=NR',-S(O)R',-SO2R',-SO2NR'R",-NR"SO2R,-CN和-NO2,-N3,-CH(Ph)2，全氟(C1-C4)烷氧基和全氟(C1-C4)烷基，取代基個數從0到3，優選0,1或者2個取代基。R',R"和R'"各自獨立選自氫，未取代的(C1-C8)的烷基和雜烷基，未取代的芳基，含有1到3個鹵素取代的芳基，未取代的烷基，烷氧基或者硫代烷基，或者芳基-(C1-C4)烷基。當R'和R"連接在相同的氮原子上時，它們能夠連同氮原子形成5-，6-或者7-元環。因此，-NR'R"包括1-吡咯烷基和4-嗎啉基，"烷基”包括例如三鹵代烷基(如，-CF3和-CH2CF3)，且當芳基是四氫化萘時，它可以被取代或者未取代的(C3-C7)螺環烷基所取代。(C3-C7)螺環烷基可以按照本發明定義“環烷基”的方式被取代。 Substituents are selected from: halogen, -R', -OR', =O, =NR', =N-OR', -NR'R", -SR', -SiR'R"R'", -OC( O) R', -C(O)R', -CO 2 R', -CONR'R", -OC(O)NR'R", -NR"C(O)R', -NR'-C (O)NR"R'",-NR'-SO 2 NR'",-NR"CO 2 R',-NH-C(NH 2 )=NH,-NR'C(NH 2 )=NH,- NH-C(NH 2 )=NR', -S(O)R', -SO 2 R', -SO 2 NR'R", -NR"SO 2 R, -CN and -NO 2 , -N 3 , -CH(Ph) 2 , perfluoro(C1-C4)alkoxy and perfluoro(C1-C4)alkyl, the number of substituents being from 0 to 3, preferably 0, 1 or 2 substituents. R', R" and R'" are each independently selected from hydrogen, unsubstituted (C1-C8) alkyl and heteroalkyl, unsubstituted aryl, containing 1 to 3 halogen-substituted aryl, unsubstituted Alkyl, alkoxy or thioalkyl, or aryl-(C1-C4)alkyl. When R' and R" are attached to the same nitrogen atom, they can form a 5-, 6- or 7-membered ring along with the nitrogen atom. Thus, -NR'R" includes 1-pyrrolidinyl and 4-morpholine The "alkyl group" includes, for example, a trihaloalkyl group (e.g., -CF 3 and -CH 2 CF 3 ), and when the aryl group is tetralin, it may be substituted or unsubstituted (C3-C7) spiro ring. Substituted by an alkyl group. The (C3-C7)spirocycloalkyl group may be substituted in the manner of "cycloalkyl" as defined herein.
優選的取代基選自：鹵素，-R',-OR',=O,-NR'R",-SR',-SiR'R"R'",-OC(O)R',-C(O)R',-CO2R',-CONR'R",-OC(O)NR'R",-NR"C(O)R',-NR"CO2R',-NR'-SO2NR"R'",- S(O)R',-SO2R',-SO2NR'R",-NR"SO2R,-CN和-NO2，全氟(C1-C4)烷氧基和全氟(C1-C4)烷基，其中R'和R"如上所述。 Preferred substituents are selected from the group consisting of: halogen, -R', -OR', =O, -NR'R", -SR', -SiR'R"R'", -OC(O)R', -C( O) R', -CO 2 R', -CONR'R", -OC(O)NR'R", -NR"C(O)R', -NR"CO 2 R',-NR'-SO 2 NR"R'",- S(O)R',-SO 2 R',-SO 2 NR'R",-NR"SO 2 R,-CN and -NO 2 ,perfluoro(C1-C4) Alkoxy and perfluoro(C1-C4)alkyl, wherein R' and R" are as described above.
這裡的術語“稠環”指的是一類多環體系，例如，雙環或三環體系，其中兩個環僅共用兩個環原子和一個化學鍵。稠環的例子可以包括稠合的雙環烷基環，所述雙環烷基環由7到12個環原子排列成的雙環，所述雙環選自如上所述的[4,4],[4,5],[5,5],[5,6]和[6,6]環體系；稠合雙芳香環，例如如上所述的7到12元雙芳香環體系，稠合三芳香環，例如如上所述的10到15元三環芳香環體系；稠合的雙雜芳環，例如如上所述的8-到12-元雙雜芳香環體系，稠合的三雜芳環，例如如上所述的11-到14-元三環雜芳香環體系；以及如上所述的稠合雙環或三環雜芳環。 The term "fused ring" as used herein refers to a class of polycyclic systems, for example, bicyclic or tricyclic systems in which two rings share only two ring atoms and one chemical bond. Examples of the fused ring may include a fused bicycloalkyl ring which is a bicyclic ring in which 7 to 12 ring atoms are arranged, and the bicyclo ring is selected from [4, 4], [4, as described above. 5], [5,5], [5,6] and [6,6] ring systems; fused diaromatic rings, such as the 7 to 12 membered biaromatic ring system described above, fused triaromatic rings, for example a 10 to 15 membered tricyclic aromatic ring system as described above; a fused biheteroaryl ring such as an 8- to 12-membered diheteroaromatic ring system as described above, a fused triheteroaryl ring, for example as described above An 11- to 14-membered tricyclic heteroaromatic ring system; and a fused bicyclic or tricyclic heteroaryl ring as described above.
這裡所述的化合物可以含有一個不對稱中心，因而可以作為對映異構體存在。這裡所述的化合物具有兩個或多個不對稱中心，它們另外可以作為非對映異構體存在。對映異構體和非對映異構體屬於廣泛的一類立體異構體。所有這些可能的立體異構體包括大體上純的拆分的對映異構體，外消旋混合物，和非對映體混合物。本文所公開的所述化合物的全部立體異構體和/或其中的藥學上可接受的鹽類都包括在內。除非另外的特別提到，否則提到的一個異構體適用於任何一個合理的異構體。無論何時同分異構組分未指明的，所有可能的同分異構體都包括在內。 The compounds described herein may contain an asymmetric center and thus may exist as enantiomers. The compounds described herein have two or more asymmetric centers which may additionally exist as diastereomers. Enantiomers and diastereomers belong to a broad class of stereoisomers. All such possible stereoisomers include substantially pure resolved enantiomers, racemic mixtures, and diastereomeric mixtures. All stereoisomers of the compounds disclosed herein and/or pharmaceutically acceptable salts thereof are included. Unless otherwise specifically mentioned, one of the isomers mentioned applies to any reasonable isomer. All possible isomers are included whenever the isomeric component is not indicated.
術語"大體上純的"意思是目標立體異構體所包含其它的立體異構體的重量不超過35%，比如不超過30%，更進一步不超 過25%，甚至不超過20%。在一些實施例中，術語"大體上純的"意思是目標立體異構體所包含其它立體異構體的重量不超過10%，例如不超過5%，比如不超過1%。 The term "substantially pure" means that the other stereoisomer contained in the target stereoisomer has a weight of no more than 35%, such as no more than 30%, and further does not exceed Over 25%, or even no more than 20%. In some embodiments, the term "substantially pure" means that the other stereoisomers contained in the target stereoisomer have a weight of no more than 10%, such as no more than 5%, such as no more than 1%.
當一些化合物包含烯族雙鍵時，除非另外詳細的說明，否則這些雙鍵包括E和Z式幾何異構體。 When some compounds contain olefinic double bonds, these double bonds include the E and Z geometric isomers unless otherwise specified.
一些化合物可以存在不同的氫原子連接點，被稱為互變異構體。例如，包括羰基-CH2C(O)-基團(酮式)的化合物可以經歷互變異構形成羥基-CH=C(OH)-基團(烯醇式)。酮式和烯醇式在使用時，單一形式以及混合物也包括在內。 Some compounds may have different hydrogen atom attachment points and are referred to as tautomers. For example, a compound comprising a carbonyl-CH 2 C(O)- group (keto form) can undergo tautomerization to form a hydroxy-CH=C(OH)- group (enol form). The keto and enol forms, when used, are also included in a single form as well as mixtures.
將反應產物彼此分離，或者跟原料分開是有利的。每一步或一連串幾步的目標物被分離和/或純化(下文中使用分離)通過本領域中常用的技術達到渴望的均勻度。代表性的分離技術包括多相提取，用一種溶劑或混合溶劑重結晶，蒸餾，昇華，或色譜法。色譜法可以涉及到許多方法，包括例如：反相和正相；分子篩，離子交換，高，中，低壓液相色譜法和設備；小型分析(small scale analytical)；模擬化移動床("SMB")和製備薄層或厚層層析法，和小型薄層和快速色譜技術一樣。本領域技術人員使用這些技術達到所需分離度。 It is advantageous to separate the reaction products from each other or separately from the starting materials. Each step or series of steps of the target is separated and/or purified (using separations hereinafter) to achieve the desired uniformity by techniques commonly used in the art. Representative separation techniques include multiphase extraction, recrystallization with a solvent or mixed solvent, distillation, sublimation, or chromatography. Chromatography can involve a number of methods including, for example, reversed phase and normal phase; molecular sieves, ion exchange, high, medium, low pressure liquid chromatography and equipment; small scale analytical; simulated moving bed ("SMB") And preparation of thin layer or thick layer chromatography, as well as small thin layer and flash chromatography techniques. Those skilled in the art use these techniques to achieve the desired resolution.
非對映異構體的混合物可以利用它們物理化學的差異通過本領域公知的技術被分離成各自的非對映異構體，比如通過色譜法和/或分步結晶法。對映異構體能夠通過將對映異構體混合物與合適的光學活性化合物(例如，手性助劑如手性醇或Mosher醯氯) 反應轉化為非對映異構體的混合物，然後將非對映異構體的混合物分離，並將各個非對映異構體轉化(如水解)為相應的純對映異構體。對映異構體還能夠用手性HPLC柱分離。 Mixtures of diastereomers can be separated into the individual diastereomers by techniques which are well known in the art using physicochemical differences, such as by chromatography and/or fractional crystallization. The enantiomers can be obtained by combining the enantiomeric mixture with a suitable optically active compound (for example, a chiral auxiliary such as a chiral alcohol or Mosher® chlorine) The reaction is converted to a mixture of diastereomers, then the mixture of diastereomers is separated and the individual diastereomers are converted (e.g., hydrolyzed) to the corresponding pure enantiomers. The enantiomers can also be separated by a chiral HPLC column.
單一的立體異構體(例如大體上純的對映異構體)可以通過拆分外消旋混合物的方法獲得，比如利用光學活性的拆分劑形成非對映體的方法(Eliel,E.和Wilen,S.Stereochemistry of Organic Compounds.New York：John Wiley & Sons,Inc.,1994；Lochmuller,C.H.,等人."Chromatographic resolution of enantiomers：Selective review." J.Chromatogr.,113(3)(1975)：pp.283-302)。本發明的手性化合物的外消旋混合物可以通過任何適合的方法來分離，包括：(1)與手性化合物形成離子的，非對映異構的鹽，然後通過分步結晶或其它方法分離，(2)與手性衍生試劑形成非對映異構化合物，分離形成的非對映異構體以及轉化成純的立體異構體，(3)直接在手性條件下分離大體上純或富含的立體異構體。參見：Wainer,Irving W.,Ed.Drug Stereochemistry：Analytical Methods and Pharmacology.New York：Marcel Dekker,Inc.,1993。 Single stereoisomers (e.g., substantially pure enantiomers) can be obtained by resolution of the racemic mixture, such as by the use of optically active resolving agents to form diastereomers (Eliel, E. And Wilen, S. Stereochemistry of Organic Compounds. New York: John Wiley & Sons, Inc., 1994; Lochmuller, CH, et al. "Chromatographic resolution of enantiomers: Selective review." J. Chromatogr., 113(3) ( 1975): pp.283-302). The racemic mixture of the chiral compound of the present invention can be isolated by any suitable method, including: (1) formation of an ionic, diastereomeric salt with a chiral compound, followed by separation by fractional crystallization or other methods. (2) forming a diastereomeric compound with a chiral derivatizing reagent, separating the diastereomer formed and converting into a pure stereoisomer, and (3) separating substantially pure or under chiral conditions. Enriched stereoisomers. See: Wainer, Irving W., Ed. Drug Stereochemistry: Analytical Methods and Pharmacology. New York: Marcel Dekker, Inc., 1993.
“藥學上可接受的鹽”包括但不局限於無機酸鹽，選自比如，鹽酸鹽，磷酸鹽，磷酸氫鹽，氫溴酸鹽，硫酸鹽，亞硫酸鹽和硝酸鹽；也包括有機鹽，選自例如蘋果酸鹽，馬來酸鹽，延胡索酸鹽，酒石酸鹽，琥珀酸鹽，檸檬酸鹽，乳酸鹽，甲磺酸鹽，對甲苯磺酸鹽，2-羥基乙基磺酸鹽，苯甲酸鹽，水楊酸鹽，硬脂酸鹽， 鏈烷酸鹽比如乙酸鹽，和HOOC-(CH2)n-COOH的鹽，這裡的n選自0到4。類似地，藥學中可接受的陽離子的例子包括但不限於鈉鹽，鉀鹽，鈣鹽，鋁鹽，鋰鹽和銨鹽。 "Pharmaceutically acceptable salts" include, but are not limited to, mineral acid salts selected from, for example, hydrochlorides, phosphates, hydrogen phosphates, hydrobromides, sulfates, sulfites, and nitrates; a salt selected from, for example, malate, maleate, fumarate, tartrate, succinate, citrate, lactate, methanesulfonate, p-toluenesulfonate, 2-hydroxyethylsulfonate a salt of a benzoate, a salicylate, a stearate, an alkanoate such as an acetate, and a salt of HOOC-(CH 2 ) n -COOH, wherein n is selected from 0 to 4. Similarly, examples of pharmaceutically acceptable cations include, but are not limited to, sodium, potassium, calcium, aluminum, lithium and ammonium salts.
另外，如果本文中所述的一個化合物得到其酸加成鹽，其遊離堿可以通過鹼化其鹽溶液來得到。相反地，如果這個化合物是遊離堿，加成鹽(例如藥學可接受的加成鹽)可以通過將遊離堿溶於一種合適的有機溶劑並且用酸處理其溶液的方法製作，與由鹼性化合物製備酸加成鹽的常規程式一致。所屬領域技術人員會識別各種合成方法，所述合成方法不需過度的實驗就可以用於製備無毒的藥學可接受的加成鹽。 Further, if a compound described herein obtains its acid addition salt, its free hydrazine can be obtained by alkalizing its salt solution. Conversely, if the compound is a free oxime, an addition salt (e.g., a pharmaceutically acceptable addition salt) can be prepared by dissolving the free hydrazine in a suitable organic solvent and treating the solution with an acid, with a basic compound. The conventional procedure for preparing acid addition salts is consistent. Those skilled in the art will recognize various synthetic methods that can be used to prepare non-toxic pharmaceutically acceptable addition salts without undue experimentation.
"治療(Treating)"，"治療(treat)"，或者治療(treatment)"指的是施用至少一個化合物和/或其中至少一個立體異構體，和/或至少一個本文公開的藥學可接受的鹽給確認需要其的受試者，例如，所述受試者患有癌症。 "Treating", "treat", or "treatment" refers to the administration of at least one compound and/or at least one of its stereoisomers, and/or at least one pharmaceutically acceptable as disclosed herein. The salt is given to a subject who is in need of confirmation, for example, the subject has cancer.
“有效量”是指至少一種化合物和/或至少一種其立體異構體，和/或至少一種藥學上可接受的鹽的量，該量能夠有效地“治療”受試者的疾病或病症，而且在一定程度上，將會看到組織，系統，動物或者人類在生物學或者藥學上的反應，例如一個或多個症狀或者正在接受治療的疾病，當給藥時，足以阻止其發展，或者在一定程度上的減輕。根據化合物，疾病及其嚴重程度，年齡，體重等，待治療的哺乳動物的治療有效量會有所不同。 "Effective amount" means an amount of at least one compound and/or at least one of its stereoisomers, and/or at least one pharmaceutically acceptable salt, which amount is effective to "treat" a disease or condition in a subject, And to a certain extent, will see a biological or pharmacy response of a tissue, system, animal or human, such as one or more symptoms or a disease being treated, when administered, sufficient to prevent its development, or Reduced to a certain extent. The therapeutically effective amount of the mammal to be treated will vary depending on the compound, the disease and its severity, age, weight, and the like.
本文所述的術語“至少一個取代基”包括，例如，從1到 4，例如從1到3，進一步例如從1到2個取代基。例如本文所述“至少一個取代基R16”包括從1到4，例如從1到3，進一步例如從1到2個選自本文所述R16列表中的取代基。 The term "at least one substituent" as used herein includes, for example, from 1 to 4, such as from 1 to 3, and further, for example, from 1 to 2 substituents. For example herein, the "at least one substituent R & lt 16" comprises from 1 to 4, for example, from 1 to 3, further such as 16 from the list of substituents R 1 to 2 substituents selected article.
本發明提供了選自式中的化合物：
及其立體異構體和它們在藥學上可接受的鹽。 And stereoisomers thereof and their pharmaceutically acceptable salts.
R1(和R4，R5，R6和R7)分別獨立地選自氫，鹵素，雜烷基，烷基，烯基，環烷基，芳基，飽和或者不飽和的雜環基，雜芳基，炔基，-CN,-NR13R14,-OR13,-COR13,-CO2R13,-CONR13R14,-C(=NR13)NR14R15,-NR13COR14,-NR13CONR14R15,-NR13CO2R14,-SO2R13,-NR13SO2NR14R15或者-NR13SO2R14，其中，該烷基，烯基，炔基，環烷基，雜芳基，芳基和飽和或者不飽和的雜環基可以視需要被至少一個取代基R16任意取代。 R 1 (and R 4 , R 5 , R 6 and R 7 ) are each independently selected from hydrogen, halogen, heteroalkyl, alkyl, alkenyl, cycloalkyl, aryl, saturated or unsaturated heterocyclic groups. , heteroaryl, alkynyl, -CN, -NR 13 R 14 , -OR 13 , -COR 13 , -CO 2 R 13 , -CONR 13 R 14 , -C(=NR 13 )NR 14 R 15 ,- NR 13 COR 14 , -NR 13 CONR 14 R 15 , -NR 13 CO 2 R 14 , -SO 2 R 13 , -NR 13 SO 2 NR 14 R 15 or -NR 13 SO 2 R 14 , wherein the alkyl group The alkenyl group, the alkynyl group, the cycloalkyl group, the heteroaryl group, the aryl group and the saturated or unsaturated heterocyclic group may be optionally substituted with at least one substituent R 16 as needed.
R13，R14和R15，分別獨立地選自氫，雜烷基，烷基，烯基，炔基，環烷基，飽和或者不飽和的雜環基，芳基或者雜芳基；其中(R13和R14)，和/或(R14和R15)連同它們所連接的原子，每一個可以形成一個環，該環選自視需要至少被一個取代基R16任意取代的環烷基，飽和或者不飽和的雜環，芳基和雜芳環； 在一些具體的實施例中，R13，R14和R15分別獨立地選自氫，低級烷基，或低級烷氧基。 R 13 , R 14 and R 15 are each independently selected from the group consisting of hydrogen, heteroalkyl, alkyl, alkenyl, alkynyl, cycloalkyl, saturated or unsaturated heterocyclic, aryl or heteroaryl; (R 13 and R 14 ), and/or (R 14 and R 15 ) together with the atoms to which they are attached, each of which may form a ring selected from naphthenes optionally substituted at least by one substituent R 16 Heterocyclic, saturated or unsaturated heterocyclic, aryl and heteroaryl rings; In some specific embodiments, R 13 , R 14 and R 15 are each independently selected from hydrogen, lower alkyl, or lower alkoxy.
R16選自鹵素，取代或未取代的烷基，取代或未取代的烯基，取代或未取代的炔基，取代或未取代的環烷基，取代或未取代的芳基，取代或未取代的雜芳基，取代或未取代的飽和或者不飽和的雜環基，含氧的，-CN,-OR’,-NR’R”,-COR’,-CO2R’,-CONR’R”,-C(=NR’)NR”R''',-NR’COR”,-NR’CONR’R”,-NR’CO2R”,-SO2R’,-SO2aryl,-NR’SO2NR”R'''或者NR’SO2R”，其中，R’,R”和R'''各自獨立地選自氫，鹵素，取代或未取代的烷基，取代或未取代的烯基，取代或未取代的炔基，取代或未取代的環烷基，取代或未取代的芳基，取代或未取代的雜芳基，取代或未取代的飽和或者不飽和的雜環基，其中(R’和R”)，和/或者(R”和R''')連同它們所連接的原子，可以形成一個環，該環選自環烷基，飽和或者不飽和的雜環，芳基和雜芳環；在一些具體的實施例中，在R1上的R16選自鹵素，低級烷基，或低級烷氧基。 R 16 is selected from halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or not Substituted heteroaryl, substituted or unsubstituted saturated or unsaturated heterocyclic group, oxygenated, -CN, -OR', -NR'R", -COR', -CO 2 R', -CONR'R",-C(=NR')NR"R''',-NR'COR",-NR'CONR'R",-NR'CO 2 R", -SO 2 R', -SO 2 aryl, -NR'SO 2 NR"R'"' or NR'SO 2 R", wherein R', R" and R'" are each independently selected from hydrogen, halogen, substituted or unsubstituted alkyl, substituted or Unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted saturated or unsaturated a heterocyclic group, wherein (R' and R"), and/or (R" and R''') together with the atom to which they are attached, may form a ring selected from cycloalkyl, saturated or unsaturated. heterocyclyl, aryl and heteroaryl ring; in some embodiments, R 16 selected from halogen of R 1, lower alkyl, or lower Alkoxy.
在一些具體的實施例中，R1是任選的雜-，任選取代的烴類，該烴類選自雜烷基，烷基，烯基，環烷基，芳基，飽和或不飽和的雜環基，雜芳基，其中，烷基，烯基，炔基，環烷基，雜芳基，芳基，飽和或不飽和的雜環基可以視需要被至少一個取代基R16任意取代，其中環狀結構優選3-8-元環結構，包括0-3個N， S或O雜原子，以及芳基優選5-或6-元芳香環，包括0-3個N，S或O雜原子，其中，烴類是優選C1-C12或C1-C8烴。 In some specific embodiments, R 1 is an optional hetero-, optionally substituted hydrocarbon selected from heteroalkyl, alkyl, alkenyl, cycloalkyl, aryl, saturated or unsaturated Heterocyclic group, heteroaryl group, wherein alkyl, alkenyl, alkynyl, cycloalkyl, heteroaryl, aryl, saturated or unsaturated heterocyclic group may be optionally substituted with at least one substituent R 16 Substituted wherein the cyclic structure is preferably a 3-8-membered ring structure, including 0-3 N, S or O heteroatoms, and the aryl group is preferably a 5- or 6-membered aromatic ring, including 0-3 N, S or O hetero atom, wherein the hydrocarbon is preferably a C1-C12 or C1-C8 hydrocarbon.
在一些具體的實施例中，R1選自雜烷基，烷基，烯基，環烷基，芳基，飽和或不飽和的雜環基，雜芳基，其中，烷基，烯基，炔基，環烷基，雜芳基，芳基，飽和或不飽和的雜環基可以視需要被至少一個取代基R16任意取代。 In some specific embodiments, R 1 is selected from heteroalkyl, alkyl, alkenyl, cycloalkyl, aryl, saturated or unsaturated heterocyclic, heteroaryl, wherein alkyl, alkenyl, An alkynyl group, a cycloalkyl group, a heteroaryl group, an aryl group, a saturated or unsaturated heterocyclic group may be optionally substituted with at least one substituent R 16 as needed.
在一些具體的實施例中，R1選自低級烷基或烯基，任選環的和任選取代的，特別是鹵素，低級烷基，或低級烷氧基，並且包括0-3個N，S或O雜原子。實例包括甲基環丙基，環己基，環戊基，甲氧基乙基，鹵化物，甲基，乙基，丙基和丁基。其他實例中R1部分包括5-元芳香環，如吡咯，吡唑，咪唑，呋喃和加氫類化合物(例如，吡咯烷，吡唑烷，咪唑烷，四氫呋喃)，和6-元環，如苯(苯基)，吡啶，吡喃，二嗪，三嗪類及其四嗪，和加氫類化合物(例如，環己烷，二-和四-氫吡啶，哌啶，四氫吡喃，等)，其中每一個可以被取代，特別是鹵素，低級烷基，或低級烷氧基。 In some specific embodiments, R 1 is selected from lower alkyl or alkenyl, optionally cyclic and optionally substituted, especially halogen, lower alkyl, or lower alkoxy, and includes 0-3 N, S Or O hetero atom. Examples include methylcyclopropyl, cyclohexyl, cyclopentyl, methoxyethyl, halide, methyl, ethyl, propyl and butyl. In other examples, the R 1 moiety includes a 5-membered aromatic ring such as pyrrole, pyrazole, imidazole, furan and a hydrogenated compound (for example, pyrrolidine, pyrazolidine, imidazolidine, tetrahydrofuran), and a 6-membered ring, such as Benzene (phenyl), pyridine, pyran, diazine, triazines and their tetrazines, and hydrogenated compounds (for example, cyclohexane, di- and tetra-hydropyridine, piperidine, tetrahydropyran, And the like, each of which may be substituted, especially a halogen, a lower alkyl group, or a lower alkoxy group.
L為一個鍵，CH2，NR12，O，或者S，其中R12是氫或者低級烷基，例如甲基。 L is a bond, CH 2 , NR 12 , O, or S, wherein R 12 is hydrogen or a lower alkyl group such as a methyl group.
A為一個5-或6-元芳香環，包括0-3個N，S或O雜原子。優選5-元芳香環，包括吡咯，吡唑，咪唑，呋喃和6-元環，包括苯(苯基)，吡啶，吡喃，二嗪，三嗪和四嗪類。 A is a 5- or 6-membered aromatic ring comprising 0-3 N, S or O heteroatoms. Preferred are 5-membered aromatic rings, including pyrrole, pyrazole, imidazole, furan and 6-membered rings, including benzene (phenyl), pyridine, pyran, diazine, triazine and tetrazine.
n為0，1，2，3或4，其中當n為大於1時，每個R2可以 是不同的。在具體的實施例中，n為0(即A是未取代的)。 n is 0, 1, 2, 3 or 4, wherein when n is greater than 1, each R 2 may be different. In a particular embodiment, n is 0 (ie, A is unsubstituted).
R2選自鹵素，烷基，-S-烷基，-CN,-NR13R14,-OR13,-COR13,-CO2R13,-CONR13R14,-C(=NR13)NR14R15,-NR13COR14,-NR13CONR14R15,-NR13CO2R14,-SO2R13,-NR13SO2NR14R15和-NR13SO2R14；其中R13，R14和R15，分別選自氫，雜烷基，烷基，烯基，炔基，環烷基，飽和或者不飽和的雜環基，芳基或者雜芳基；其中(R13和R14)，和/或(R14和R15)連同它們所連接的原子，每一個可以形成一個環，該環選自視需要至少被一個取代基R16任意取代的環烷基，飽和或者不飽和的雜環，芳基和雜芳環。 R 2 is selected from the group consisting of halogen, alkyl, -S-alkyl, -CN, -NR 13 R 14 , -OR 13 , -COR 13 , -CO 2 R 13 , -CONR 13 R 14 , -C(=NR 13 )NR 14 R 15 , -NR 13 COR 14 , -NR 13 CONR 14 R 15 , -NR 13 CO 2 R 14 , -SO 2 R 13 , -NR 13 SO 2 NR 14 R 15 and -NR 13 SO 2 R 14 ; wherein R 13 , R 14 and R 15 are each independently selected from the group consisting of hydrogen, heteroalkyl, alkyl, alkenyl, alkynyl, cycloalkyl, saturated or unsaturated heterocyclic, aryl or heteroaryl; Wherein (R 13 and R 14 ), and/or (R 14 and R 15 ) together with the atoms to which they are attached, each may form a ring selected from a ring optionally substituted at least by one substituent R 16 as desired. Alkyl, saturated or unsaturated heterocyclic, aryl and heteroaryl rings.
在一些具體的實施例中，R2選自鹵素，烷基，-S-烷基，-CN,-NR13R14,-OR13,-COR13,-CO2R13,-CONR13R14,-C(=NR13)NR14R15,-NR13COR14,-NR13CONR14R15,-NR13CO2R14,-SO2R13,-NR13SO2NR14R15和-NR13SO2R14；其中優選烷基(包括-S-烷基)是低級烷基，且R13，R14和R15分別獨立地選自氫，低級烷基，或低級烷氧基。 In some specific embodiments, R 2 is selected from the group consisting of halogen, alkyl, -S-alkyl, -CN, -NR 13 R 14 , -OR 13 , -COR 13 , -CO 2 R 13 , -CONR 13 R 14 , -C(=NR 13 )NR 14 R 15 , -NR 13 COR 14 , -NR 13 CONR 14 R 15 , -NR 13 CO 2 R 14 , -SO 2 R 13 , -NR 13 SO 2 NR 14 R 15 and -NR 13 SO 2 R 14 ; wherein preferably the alkyl group (including the -S-alkyl group) is a lower alkyl group, and R 13 , R 14 and R 15 are each independently selected from hydrogen, lower alkyl, or lower alkane Oxygen.
在一些具體的實施例中，R2選自鹵素，低級烷基，或低級烷氧基。 In some specific embodiments, R 2 is selected from halogen, lower alkyl, or lower alkoxy.
每個W獨立選自-CH2-或者-C(O)-，其中如果m是2,3,or 4，優選不超過一個W是羰基；S/D是一個單鍵或者雙鍵，且當是一個雙鍵時，R5和R7不存在； m是0,1,2,3或者4，優選0,1,2或者3，且在一些具體的實施例中優選0,1或者2，或者0或者1。 Each W is independently selected from -CH 2 - or -C(O)-, wherein if m is 2, 3, or 4, preferably no more than one W is a carbonyl group; S/D is a single bond or a double bond, and is a double bond, R 5 and R 7 are absent; m is 2, 3 or 4, preferably 0,1, 2 or 3, and in some embodiments preferably 0, 1 or 2, Or 0 or 1.
p是0(且R6和R7不存在)，或1-2的整數，其中當p為0時，m是非零的，並且當p是大於1時，每個R6和R7可能是不同的；一般p+m是1,2,3或者4，優選1,2或者3，在一些實施例中優選1。 p is 0 (and R 6 and R 7 are absent), or an integer from 1 to 2, wherein m is non-zero when p is 0, and each p 6 and R 7 may be when p is greater than 1. Different; generally p+m is 1, 2, 3 or 4, preferably 1, 2 or 3, preferably 1 in some embodiments.
在一些實施例中，p是2，且m是0或者1；p是1，且m是0或者1(或者0,1或者2)；或者p是0，且m是1或者2(或者1，2或者3)。 In some embodiments, p is 2, and m is 0 or 1; p is 1, and m is 0 or 1 (or 0, 1 or 2); or p is 0, and m is 1 or 2 (or 1 , 2 or 3).
R4和R5，或R4和R6，或R6和R7，或R6和R6(當p為2時)，連同它們所連接的原子，形成一個環，該環選自視需要至少被一個取代基R16任意取代的環烷基，飽和或者不飽和的雜環，芳基和雜芳環；這些環一般為4-8-元環，包括5-元芳香環，如吡咯，吡唑，咪唑，呋喃和加氫類化合物(例如，吡咯烷，吡唑烷，咪唑烷，四氫呋喃)，和6-元環，如苯(苯基)，吡啶，吡喃，二嗪類，三嗪類及四嗪類，和加氫類化合物(例如，環己烷，二-和四-氫吡啶，哌啶，四氫吡喃，等)，其中每一個可以是未取代或者取代的，特別是鹵素，低級烷基，低級烷氧基，-COR’或者NR’COR”，其中R’,R”是取代或者未取代的烯基。 R 4 and R 5 , or R 4 and R 6 , or R 6 and R 7 , or R 6 and R 6 (when p is 2), together with the atom to which they are attached, form a ring which is selected from the group consisting of A cycloalkyl group which is optionally substituted by at least one substituent R 16 , a saturated or unsaturated heterocyclic ring, an aryl group and a heteroaryl ring; these rings are generally a 4-8-membered ring, including a 5-membered aromatic ring such as pyrrole , pyrazole, imidazole, furan and hydrogenated compounds (for example, pyrrolidine, pyrazolidine, imidazolidine, tetrahydrofuran), and 6-membered rings, such as benzene (phenyl), pyridine, pyran, diazines, Triazines and tetrazines, and hydrogenated compounds (eg, cyclohexane, di- and tetra-hydropyridine, piperidine, tetrahydropyran, etc.), each of which may be unsubstituted or substituted, In particular, halogen, lower alkyl, lower alkoxy, -COR' or NR'COR", wherein R', R" is a substituted or unsubstituted alkenyl group.
R4和R5(或R6和R7，或R6和R6，當p為2時)，在具體的實施例中，形成哌啶，氮雜環戊烷，或氮雜環丁烷，視需要任 選取代的，尤其是N-上被如下基團取代，如苄基，醯基，丙烯醯基等。 R 4 and R 5 (or R 6 and R 7 , or R 6 and R 6 , when p is 2), in a particular embodiment, form piperidine, aziridine, or azetidine Optionally substituted, especially on N-, substituted by a group such as benzyl, fluorenyl, acryloyl, and the like.
在一些具體的實施例中，R4和R6，與它們所連接的原子，形成一個式中的環：其中：Q是-CH2；J是-CH2-；且d和b各自獨立地選自0，或1-4的整數。 In some specific embodiments, R 4 and R 6 , with the atoms to which they are attached, form a ring in the formula: Wherein: Q is -CH 2; J is -CH 2 -; and d is an integer of 0, or 1, and b are each independently selected.
在一些具體的實施例中，R4和R6形成苯基，哌啶，氮雜環庚烷，吡咯烷，視需要任選取代的，尤其是N-上被如下基團取代，如苄基，醯基，丙烯醯基，甲胺-丙烯醯基等。 In some specific embodiments, R 4 and R 6 form phenyl, piperidine, azepane, pyrrolidine, optionally substituted, especially on N-, substituted by a group such as benzyl , mercapto, acrylonitrile, methylamine-acrylonitrile, and the like.
本發明包括詳細列舉的和優選方案的所有組合，就像每個組合分別進行了逐個列舉。例如，在具體實施例中，上文中的A為苯基；W是-(CH2)-；L為O；S/D是單鍵；m為1；n為0；p是1；R1是苯基；R2不存在；R5是H；以及R6和R7是H；所得到的組合。 The present invention includes all combinations of the detailed enumerated and preferred embodiments, as each combination is recited individually. For example, in a particular embodiment, the above A is phenyl; W is - (CH 2) -; L is O; S / D is a single bond; m is 1; n is 0; p is 1; R 1 Is a phenyl group; R 2 is absent; R 5 is H; and R 6 and R 7 are H; the resulting combination.
上文中的R4，包括含N的C1-C8的烷基，含N的C3-C8環烷基和苯基，例如，甲胺基，苯胺基，氮雜環丁烷基，吡咯
烷基，哌啶基，氮雜環庚烷基，上述每個基團可以被任意取代，特別是N-上被下列基團取代，如苄基，醯基，丙烯醯基，取代的丙烯醯基，丙炔醯基，取代的-丙炔醯基等，如下結構的組合：
在具體實例中，R4是1-丙烯醯基哌啶-4-基(如化合物27)或1-(丁-2-炔醯基)哌啶-4-基(如化合物176)。 In a particular embodiment, R 4 is 1-propenylpiperidin-4-yl (such as compound 27) or 1-(but-2-ynindolyl)piperidin-4-yl (such as compound 176).
本發明還提供了實施例，和表I，II和III中的所有化合物，其立體異構體，及藥學上可接受的鹽。 The invention also provides examples, and all of the compounds in Tables I, II and III, stereoisomers, and pharmaceutically acceptable salts thereof.
本發明中的化合物，及其立體異構體，和其藥學上可接受的鹽，在治療中可以採用單獨使用，或者與至少一種其他治療藥物聯合使用。在一些實施例中，本發明化合物，其立體異構體，以及藥學上可接受的鹽可以與至少一種額外的治療試劑聯合使用。至少一種額外的治療試劑例如可以選自於抗增殖、抗癌症和化療試劑。本發明化合物和/或一種藥學上可以接受的鹽可以和至少一種其它的治療試劑以單一劑型或者不同的劑型聯合給藥。當不同劑型聯合給藥時，另外至少一種其它的治療試劑，可以先於給藥，同時給藥或者給完本發明中公開的化合物和/或藥學上可以接受的鹽之後給藥。 The compounds of the present invention, and stereoisomers thereof, and pharmaceutically acceptable salts thereof, may be used alone or in combination with at least one other therapeutic agent in therapy. In some embodiments, a compound of the invention, a stereoisomer thereof, and a pharmaceutically acceptable salt can be used in combination with at least one additional therapeutic agent. The at least one additional therapeutic agent can be selected, for example, from anti-proliferative, anti-cancer, and chemotherapeutic agents. The compounds of the invention and/or a pharmaceutically acceptable salt can be administered in combination with at least one other therapeutic agent in a single dosage form or in a different dosage form. When the different dosage forms are administered in combination, the additional at least one additional therapeutic agent can be administered prior to administration, simultaneously or after administration of the compound and/or pharmaceutically acceptable salt disclosed herein.
“化療試劑”是一種不考慮作用機制，用於治療癌症的化合物。化療試劑可以是在“靶向治療”和常規化療中使用的化合物。合適的化療試劑可以選自例如引起細胞凋亡的試劑；多聚核苷酸(如，核酶)；多肽(如，酶)；藥物；生物模擬物；生物鹼；烷基化試劑；抗腫瘤的抗生素；抗代謝物；激素；鉑化合物；與抗癌藥物，毒素，和/或者放射性核素連用的單克隆抗體；生物應答調節劑(幹擾素，比如IFN-a和白細胞介素，比如IL-2)；過繼免疫治療劑；造血生長因數；引起腫瘤細胞分化的試劑(如全反式-維甲酸)；基因治療試劑；反義治療試劑和核苷；腫瘤疫苗；和一些血管再生抑制劑。 A "chemotherapeutic agent" is a compound that is used to treat cancer without regard to the mechanism of action. The chemotherapeutic agent can be a compound used in "targeted therapy" and conventional chemotherapy. Suitable chemotherapeutic agents can be selected, for example, from agents that cause apoptosis; polynucleotides (eg, ribozymes); polypeptides (eg, enzymes); drugs; biomime; alkaloids; alkylating agents; Antibiotics; antimetabolites; hormones; platinum compounds; monoclonal antibodies used in combination with anticancer drugs, toxins, and/or radionuclides; biological response modifiers (interferons such as IFN-a and interleukins such as IL) -2); adoptive immunotherapeutic agents; hematopoietic growth factors; agents that cause tumor cell differentiation (eg, all-trans-retinoic acid); gene therapy agents; antisense therapeutic agents and nucleosides; tumor vaccines; and some angiogenesis inhibitors .
化療藥物的例子包括厄洛替尼(特羅凱®，Genentech/OSI Pharm.)；硼替佐米(萬珂®，Millennium Pharm.)；氟維司群(FASLODEX®，阿斯利康)；舒尼替尼(索坦®，輝瑞)；來曲唑(弗隆®，諾華)；伊馬替尼甲磺酸鹽(格列衛®，諾華)；PTK787/ZK 222584(諾華)；奧沙利鉑(樂沙定®，賽諾菲)；5-FU(5-氟尿嘧啶)；亞葉酸鈣；雷帕黴素(西羅莫司，RAPAMUNE®，惠氏公司)；拉帕替尼(TYKERB ®，GSK572016，葛蘭素史克)；洛那法尼(SCH 66336)；索拉非尼(多吉美®，拜耳)；伊立替康(CAMPTOSAR®，輝瑞)和吉非替尼(易瑞沙®，阿斯利康)；AG1478，AG1571(SU 5271，Sugen)；烷基化試劑，如噻替派和CYTOXAN®環磷醯胺；烷基磺酸鹽，例如白消安，improsulfan和哌泊舒凡；氮雜環丙烷，如 benzodopa，carboquone，meturedopa和uredopa；ethylenimines和methylamelamines如六甲蜜胺，triethylenemelamine，三亞乙基磷醯胺，三亞乙基硫代磷醯胺和三羥甲蜜胺；內酯(如bullatacin和bullatacinone)；喜樹堿(比如托泊替康的合成類似物)；苔蘚抑素；callystatin；CC-1065和其阿多來新，卡折來新，比折來新的合成類似物；cryptophycins(如cryptophycin 1和cryptophycin 8)；朵拉司他汀；duocarmycin和其合成類似物，如KW-2189和CB1-TM1；eleutherobin；pancratistatin；a sarcodictyin；spongistatin；氮芥，如苯丁酸氮芥，chlomaphazine，chlorophosphamide，雌氮芥，異環磷醯胺，氮芥，氧化氮芥鹽酸鹽，馬法蘭，新恩比興(novembichin)，苯芥膽甾醇(phenesterine)，潑尼氮芥(prednimustine)，三芥環磷醯胺，尿嘧啶氮芥；亞硝基脲如卡莫司汀，氯脲黴素，福莫司汀，洛莫司汀，尼莫司汀和ranimnustine；抗生素如烯二炔類抗生素(如卡裡奇黴素gamma1I和卡裡奇黴素omegaI1(Angew Chem.Intl.Ed.Engl.(1994)33：183-186))；dynemicin，比如dynemicin A；二膦酸鹽，如氯膦酸鹽；埃斯培拉黴素；和新制癌菌素髮色團，以及相關的生色烯二炔類抗生素的發色團，aclacinomysins，放線菌素(actinomycin)，authramycin，重氮絲氨酸，博萊黴素，放線菌素C(cactinomycin)，carabicin，caminomycin，carzinophilin，chromomycinis，放線菌素D(dactinomycin)，，柔紅黴素(daunorubicin)；地托比星 (detorubicin)，6-重氮基-5-氧代-L-正亮氨酸，阿黴素®(多柔比星，doxorubicin)，嗎啉基-阿黴素，氰基嗎啉基-阿黴素，2-pyrrolino-doxorubicin和去氧阿黴素，表柔比星(epirubicin)，依索比星(esorubicin)，伊達比星(idarubicin)，麻西羅黴素(marcellomycin)，絲裂黴素如絲裂黴素C，黴酚酸，諾加黴素(nogalamycin)，橄欖黴素(olivomycins)，培洛黴素，泊非黴素(porfiromycin)，嘌呤黴素，quelamycin，羅多比星(rodorubicin)，鏈黑菌素，鏈脲菌素，殺結核菌素(tubercidin)，烏苯美司，淨司他丁(zinostatin)，佐柔比星(zorubicin)；抗代謝產物如氨甲喋呤和5-氟尿嘧啶(5-FU)；葉酸類似物如二甲葉酸(denopterin)，氨甲喋呤，蝶羅呤，三甲曲沙；嘌呤類似物如氟達拉濱，6-巰基嘌呤，硫咪嘌呤(thiamiprine)，硫鳥嘌呤；嘧啶類似物如環胞苷，阿紮胞苷，6-氮雜尿苷(6-azauridine)，卡莫氟，阿糖胞苷，二去氧尿苷，去氧氟尿苷，依諾他濱，氟尿苷；雄激素，如卡普睾酮(calusterone)，屈他雄酮丙酸酯(dromostanolone propionate)，環硫雄醇，美雄烷(mepitiostane)，睾內酯；抗腎上腺素如氨魯米特，米托坦，曲洛司坦；葉酸補充劑如frolinic acid；醋葡醛內酯；醛磷醯胺苷；氨基酮戊酸；eniluracil；安吖啶；bestrabucil；比生群(bisantrene)；edatraxate；defofamine；脫羰秋水仙堿(demecolcine)；地吖醌(diaziquone)；elformithine；依利醋銨(elliptinium acetate)；埃博黴素；依託格魯(etoglucid)；硝酸 鎵；羥基脲；香菇多糖；lonidainine；美登木素生物鹼(maytansinoids)，如美登素(maytansine)和安絲菌素(ansamitocins)；米托胍腙(mitoguazone)；米托蒽醌；mopidanmol；nitraerine；噴司他丁；苯來美特(phenamet)；吡柔比星；洛索蒽醌(losoxantrone)；podophyllinic acid；2-乙基醯肼；甲基苄肼；PSK®多糖複合物(JHS Natural Products,Eugene,Oreg.)；雷佐生(razoxane)；根黴素(rhizoxin)；sizofuran；鍺螺胺(spirogermanium)；細交鏈孢菌酮酸(tenuazonic acid)；三亞胺醌(triaziquone)；三(2-氯乙基)胺；單端孢素(如T-2毒素，verracurin A，漆斑菌素A(roridin A)和anguidine)；氨基甲酸乙酯；長春地辛；氮烯咪胺；甘露醇氮芥；二溴甘露醇(mitobronitol)；二溴衛矛醇(mitolactol)；胍血生(pipobroman)；gacytosine；***糖苷("Ara-C")；環磷醯胺；塞替派；紫杉烷類，例如，TAXOL®(紫杉醇；Bristol-Myers Squibb Oncology,Princeton,N.J.)，ABRAXANE®(Cremophor-free)，白蛋白工程紫杉醇納米顆粒製劑(American Pharmaceutical Partners,Schaumberg,Ill.)，和泰索帝®(doxetaxel；Rhone-Poulenc Rorer,Antony,France)；chloranmbucil；健擇®(吉西他濱)；6-硫代鳥嘌呤；巰基嘌呤；氨甲喋呤；鉑類似物，如順鉑，卡鉑；長春花堿；依託泊苷(etoposide)(VP-16)；異環磷醯胺；米托蒽醌；長春新堿；諾維本®(長春瑞濱)；米托蒽醌(novantrone)；替尼泊苷；依達曲沙 (edatrexate)；道諾黴素；氨基蝶呤；卡培他濱(XELODA®)；伊班膦酸鈉；CPT-11；拓撲異構酶抑制劑RFS 2000；二氟甲基鳥氨酸(DMFO)；類視色素如視黃酸；藥學上可接受的鹽，酸和任何上述的衍生物。 Examples of chemotherapeutic drugs include erlotinib (Terroy®, Genentech/OSI Pharm.); bortezomib (Millennium Pharm.); fulvestrant (FASLODEX®, AstraZeneca); Tini (Soltan®, Pfizer); Letrozole (Fron®, Novartis); Imatinib mesylate (Gleevec®, Novartis); PTK787/ZK 222584 (Nova); oxaliplatin Leshadine®, Sanofi); 5-FU (5-fluorouracil); calcium leucovorin; rapamycin (sirolimus, RAPAMUNE®, Wyeth); lapartini ® (GSK572016, GlaxoSmithKline; Lonafani (SCH 66336); Sorafenib (Nexaco®, Bayer); Irinotecan (CAMPTOSAR®, Pfizer) and Gefitinib (Iressa®, AstraZeneca) AG1478, AG1571 (SU 5271, Sugen); alkylating agents such as thiotepa and CYTOXAN® cyclophosphamide; alkyl sulfonates such as busulfan, improsulfan and piperazine; nitrogen heterocycle Propane, such as benzodopa, carboquone, meturedopa and uredopa; ethylenimines and methylamelamines such as hexamethylene melamine, triethylenemelamine, triethylenephosphonium, triethylene thiophosphonamide and tris Melamine; lactones (such as bullatacin and bullatacinone); camptotheca (such as synthetic analogs of topotecan); bryostatin; callistatin; CC-1065 and its adoline, card fold new, fold New synthetic analogues; cryptophycins (such as cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin and its synthetic analogues such as KW-2189 and CB1-TM1; eleutherobin; pancratistatin; a sarcodictyin; spongistatin; Such as chlorambucil, chlomaphazine, chlorophosphamide, estramustine, ifosfamide, nitrogen mustard, nitrogen oxide mustard hydrochloride, melphalan, neombichin, phenesterine, splash Prednimustine, tri-salt cyclophosphamide, uracil mustard; nitrosourea such as carmustine, chlorflurimycin, formoterol, lomustine, nimustine and ranimnustine Antibiotics such as enediyne antibiotics (such as calicheamicin gamma1I and calicheamicin omegaI1 (Angew Chem. Intl. Ed. Engl. (1994) 33: 183-186)); dynemicin, such as dynemicin A; Bisphosphonates such as clodronate; espermati; and new systems Chromophore chromophore, and related chromophores of chromogenic diacetylene antibiotics, aclacinomysins, actinomycin, authramycin, azaserine, bleomycin, cactinomycin, carabicin , caminomycin, carzinophilin, chromomycinis, actinomycin D, daunorubicin; detorubicin, 6-diazo-5-oxo-L-norleucine, adriamycin ® (doxorubicin, doxorubicin), morpholino - doxorubicin, morpholino-cyano - doxorubicin, 2-pyrrolino-doxorubicin and deoxy doxorubicin, epirubicin (epirubicin) , esorubicin, idarubicin, marcellomycin, mitomycin such as mitomycin C, mycophenolic acid, nogalamycin, olive mold Olivomycins, piroximycin, porfiromycin, puromycin, quelamycin, rodorubicin, streptavidin, streptozotocin, tuberculin, Ubumex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5- Uracil (5-FU); folic acid analogues such as dinophoric acid (denopterin), methotrexate, pteroquinone, trimethoate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, Thioguanine; pyrimidine analogs such as cyclosporine, azacitidine, 6-azauridine, carmofur, cytarabine, di-deoxyuridine, deoxyfluorouridine, Enesitabine, fluorouridine; androgens, such as calpressone, dromostanolone propionate, cyclostrankrol, mepitiostane, testosterone; anti-adrenalin Such as ammonia, mitoxantrone, tromethamine; folic acid supplements such as frolinic acid; acetaldehyde lactone; aldoxime; amino keto valerate; eniluracil; amidine; bestrabucil; (bisantrene); edoftraxate; defofamine; dedecolamine; diaziquone; elformithine; elliptinium acetate; epothilone; etoglucid; Urea; lentinan; lonidainine; maytansinoids, such as mayon Maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentastatin; phenamet; pirarubicin; Los (losoxantrone); podophyllinic acid; 2-ethyl hydrazine; procarbazine; PSK® polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; Sizofuran; spirogermanium; tenuazonic acid; triaziquone; tris(2-chloroethyl)amine; trichosporin (eg T-2 toxin, verracurin) A, roridin A and anguidine); urethane; vindesine; azomethamine; mannitol mustard; dibromomannitol; mitolactol Pipobroman; gacytosine; arabinoside ("Ara-C");cyclophosphamide;thiotepa; taxanes, for example, TAXOL® (Paclitaxel; Bristol-Myers Squibb Oncology, Princeton, NJ ), ABRAXANE® (Cremophor-free), albumin engineered paclitaxel nanoparticle formulation (American Pharmaceutical Partners, Schaumberg, Ill) .), And TAXOTERE ® (doxetaxel; Rhone-Poulenc Rorer , Antony, France); chloranmbucil; GEMZAR ® (gemcitabine); 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin, Carboplatin; vinca sinensis; etoposide (VP-16); methotrexate; mitoxantrone; vinorelbine; noviben® (vinorelbine); mitantrone ; teniposide; edatrexate; daunorubicin; aminopterin; capecitabine (XELODA®); ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); a retinoid such as retinoic acid; a pharmaceutically acceptable salt, an acid and any of the above derivatives.
“化療試劑”也可以選自於，比如：(i)能夠調控或者抑制激素作用於腫瘤的抗激素類試劑，比如抗***和選擇性***受體調控劑(SERMs)，包括，比如它莫昔芬(包括NOLVADEX®；它莫昔芬檸檬酸鹽)，雷洛昔芬，屈洛昔芬，4-羥基他莫昔芬，曲沃昔芬，keoxifene，LY117018，奧那司酮和FARESTON®(toremifine檸檬酸鹽)；(ii)能夠抑制芳香酶的芳香酶抑制劑，能夠調節***在腎上腺中的產生，比如，4(5)-咪唑，氨魯米特，MEGASE®(甲地孕酮醋酸鹽)，AROMASIN®(依西美坦；輝瑞)，formestanie，法倔唑，RIVISOR®(伏氯唑)，FEMARA®(來曲唑；諾華)和ARIMIDEX®(anastrozole；阿斯利康)；(iii)抗雄激素比如氟他胺，尼魯米特，比卡魯胺，亮丙瑞林(leuprolide)和戈舍瑞林；和曲沙他濱(troxacitabine，一個1,3-二惡茂烷核苷胞嘧啶類似物)；(iv)蛋白激酶抑制劑；(v)脂質激酶抑制劑；(vi)反義寡核苷酸，它們能抑制信號傳導通路中基因的表達，這些基因引起一些變異細胞的增值，比如PKC-alpha,Ralf和H-Ras；(vii)核酶比如VEGF表達抑制劑(如，ANGIOZYME®)和HER2表達抑制劑；(viii)疫苗如基因治療疫苗，如ALLOVECTIN®，LEUVECTIN®，和VAXID®； PROLEUKIN® rIL-2；拓撲異構酶I抑制劑，如LURTOTECAN®；ABARELIX® rmRH；(ix)抗血管生成試劑例如貝伐單抗(AVASTIN®，Genentech)；和(x)藥學上可接受的鹽，酸及上面提到的類似物。 "Chemotherapeutic agents" may also be selected, for example, from: (i) anti-hormonal agents that modulate or inhibit the action of hormones on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), including, for example, Tamoxifen (including NOLVADEX ® ; tamoxifen citrate), raloxifene, droloxifene, 4-hydroxytamoxifen, rovaxifene, keoxifene, LY117018, onastroxone and FARESTON ® (toremifine citrate); (ii) an aromatase inhibitor capable of inhibiting aromatase, which regulates the production of estrogen in the adrenal gland, for example, 4(5)-imidazole, aminoglutethimide, MEGASE ® Progesterone acetate), AROMASIN ® (Exemestane; Pfizer), formestanie, fadrozole, RIVISOR ® (voltazole), FEMARA ® (letrozole; Novartis) and ARIMIDEX ® (anastrozole; AstraZeneca) (iii) antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide and goserelin; and troxacitabine (a 1,3-dioxin) a cycloalkane cytosine analog); (iv) a protein kinase inhibitor; (v) a lipid kinase inhibitor; (vi) an antisense oligo Nucleotide, which can inhibit the expression of genes in signaling pathways, these genes causes some added transformed cells, such as PKC-alpha, Ralf and H-Ras; (vii) ribozymes such as VEGF expression inhibitors (e.g., ANGIOZYME ®) And HER2 expression inhibitors; (viii) vaccines such as gene therapy vaccines such as ALLOVECTIN ® , LEUVECTIN ® , and VAXID ® ; PROLEUKIN ® rIL-2; topoisomerase I inhibitors such as LURTOTECAN ® ; ABARELIX ® rmRH; An anti-angiogenic agent such as bevacizumab (AVASTIN ® , Genentech); and (x) a pharmaceutically acceptable salt, an acid and the above-mentioned analogs.
“化療試劑”也可以選自於，比如，有療效的抗體比如阿侖單抗(Campath)，貝伐單抗(AVASTIN®，Genentech)；西妥昔單抗(ERBITUX®，Imclone)；帕尼單抗(VECTIBIX®，Amgen)，利妥昔單抗(RITUXAN®，Genentech/Biogen Idec)，帕妥珠單抗(OMNITARG®,2C4,Genentech)，曲妥珠單抗(HERCEPTIN®，Genentech)，托西莫單抗(Bexxar,Corixia)，和抗體藥物結合物，吉妥珠單抗奧唑米星(MYLOTARG®，Wyeth)。 "Chemotherapeutic agents" may also be selected, for example, from therapeutic antibodies such as alemtuzumab (Campath), bevacizumab (AVASTIN ® , Genentech), cetuximab (ERBITUX ® , Imclone); Pani Monoclonal antibody (VECTIBIX ® , Amgen), rituximab (RITUXAN®, Genentech/Biogen Idec), pertuzumab (OMNITARG ® , 2C4, Genentech), trastuzumab (HERCEPTIN ® , Genentech), Toximozumab (Bexxar, Corixia), and antibody drug conjugate, gemtuzumab olozomethine (MYLOTARG ® , Wyeth).
有潛在療效的人源化單克隆抗體作為化療試劑可以和本發明中的化合物，及其異構體，和藥學上可以接受的鹽聯合用藥，比如，可以選自於：阿侖單抗，阿泊珠單抗，阿塞珠單抗，atlizumab，bapineuzumab，貝伐單抗，bivatuzumab mertansine，cantuzumab mertansine，cedelizumab，賽妥珠單抗注射液，cidfusituzumab,cidtuzumab，達利珠單抗，依庫麗單抗(eculizumab)，依法利珠單抗，依帕珠單抗(epratuzumab)，erlizumab(厄利珠單抗)，felvizumab(泛維珠單抗)，fontolizumab，吉妥珠單抗奧唑米星，inotuzumab ozogamicin，易普利姆瑪(ipilimumab),labetuzumab，林妥珠單抗，馬妥珠單 抗(matuzumab)，美泊利單抗，motavizumab，motovizumab，那他珠單抗，尼妥珠單抗，nolovizumab,numavizumab，ocrelizumab，奧馬珠單抗，帕利珠單抗，帕考珠單抗(pascolizumab)，pecfusituzumab，pectuzumab，帕妥珠單抗，pexelizumab,ralivizumab，雷珠單抗，reslivizumab,reslizumab,resyvizumab，羅維珠單抗(rovelizumab)，盧利珠單抗(ruplizumab)，西羅珠單抗(sibrotuzumab)，希普利珠單抗(siplizumab)，索土珠單抗，替他珠單抗(tacatuzumab tetraxetan)，他度珠單抗(tadocizumab)，他利珠單抗，tefibazumab，托珠單抗，toralizumab，曲妥珠單抗，tucotuzumab celmoleukin,tucusituzumab,umavizumab,urtoxazumab和維西珠單抗(visilizumab)。 A potentially potent humanized monoclonal antibody can be used as a chemotherapeutic agent in combination with a compound of the present invention, and an isomer thereof, and a pharmaceutically acceptable salt, for example, selected from the group consisting of: alemtuzumab, Pertuzumab, acezumab, atlizumab, bapineuzumab, bevacizumab, bivatuzumab mertansine, cantuzumab mertansine, cedelizumab, certolizumab injection, cidfusituzumab, cidtuzumab, daclizumab, eculizumab (eculizumab), ezetuzumab, epratuzumab, erlizumab (ehelizumab), felvizumab (panvizumab), fontolizumab, gemtuzumab ozogamicin, inotuzumab Ozogamicin, ipilimumab, labetuzumab, lintuzumab, mattobe Anti-matuzumab, mepolizumab, motavizumab, motovizumab, natalizumab, nimotuzumab, nolovizumab, numavizumab, ocrelizumab, omalizumab, palivizumab, paclizumab ( Pascolizumab), pecfusituzumab, pectuzumab, pertuzumab, pexelizumab, ralivizumab, ranibizumab, reslivizumab, reslizumab, resyvizumab, rovelizumab, ruplizumab, sibrotuzumab , Siplizumab, sibutizumab, tacilizumab tetraxetan, tadocizumab, telibizumab, tefibazumab, tocilizumab, toralizumab , trastuzumab, tucotuzumab celmoleukin, tucusituzumab, umavizumab, urtoxazumab and visilizumab.
也提供了一種藥物組合包含一種本發明中的化合物，其異構體，和藥學上可以接受的鹽，以及至少一種藥學上可以接受的載體。 Also provided is a pharmaceutical combination comprising a compound of the invention, an isomer thereof, and a pharmaceutically acceptable salt, and at least one pharmaceutically acceptable carrier.
所述藥物組合物包含一種本發明中的化合物，及其異構體，和藥學上可以接受的鹽，可以用各種已知的方式給藥，比如口服，外塗，直腸給藥，非腸道給藥，吸入噴霧，或者通過植入型藥盒，儘管在某種假定的情況下最適合的給藥途徑取決於特定的宿主，和活性成分給藥時的自然條件和疾病的嚴重程度。這裡使用的“非腸道給藥”包括皮下，皮內，靜脈注射，肌肉，關節內，動脈內，滑膜內，胸骨內，鞘內，病灶內和顱內注射或者輸液技術。本 文所公開的組合物可方便地以單位劑量的形式，並且由任何在本領域中公知的方法製備。 The pharmaceutical composition comprises a compound of the present invention, and an isomer thereof, and a pharmaceutically acceptable salt, which can be administered in various known manners, such as oral, topical, rectal, parenteral. Administration, inhalation spray, or by implantable kit, although the most appropriate route of administration under certain hypothetical circumstances depends on the particular host, and the natural conditions and severity of the disease at which the active ingredient is administered. "Parenteral administration" as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intra-arterial, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques. this The compositions disclosed herein are conveniently prepared in unit dosage form and by any methods known in the art.
本發明中的化合物，及其異構體，和藥學上可以接受的鹽能夠以固體劑型進行口服，比如膠囊，藥片，片劑，糖衣片，顆粒和粉末，或者以液體劑型進行口服，比如酏劑，糖漿，乳劑，分散液和懸浮液。本發明公開的化合物及其異構體，和藥學上可以接受的鹽也可以非腸道給藥，用無菌的液體劑型，比如分散液，懸浮液或者溶液。本發明中的化合物及其異構體，和藥學上可以接受的鹽也可以用其它的劑型來給藥比如：軟膏，乳霜，滴劑，局部用藥的透皮貼劑或者粉末，以眼用溶液或懸浮液的形式比如滴眼液，進行眼部用藥，用噴霧或者粉末組合物的形式吸入或者鼻腔給藥，或者以乳膏，軟膏，噴劑，栓劑的形式進行直腸或***給藥。 The compounds of the present invention, and isomers thereof, and pharmaceutically acceptable salts can be orally administered in solid dosage forms such as capsules, tablets, tablets, dragees, granules and powders, or orally in liquid form, such as sputum. Agents, syrups, emulsions, dispersions and suspensions. The compounds disclosed herein, and isomers thereof, and pharmaceutically acceptable salts can also be administered parenterally, in sterile liquid dosage forms such as dispersions, suspensions or solutions. The compounds of the present invention and isomers thereof, and pharmaceutically acceptable salts can also be administered in other dosage forms such as ointments, creams, drops, topical transdermal patches or powders for ophthalmic use. The solution or suspension is in the form of an eye drop, for ophthalmic administration, inhalation or nasal administration in the form of a spray or a powder composition, or rectal or vaginal administration in the form of a cream, ointment, spray, or suppository.
明膠膠囊包含本發明公開的化合物和/或者至少一種藥學上可以接受的鹽和粉末狀載體，比如乳糖，澱粉，纖維素衍生物，硬脂酸鎂，硬脂酸和相似物也可以使用。相似的稀釋劑可以用來壓片。藥片和膠囊都可以製成緩釋產品用於在一段時間裡持續給藥。壓縮的藥片可以用糖包衣或者薄膜包衣以掩蓋令人不愉快的味道，同時保護藥片隔絕空氣，或者腸溶包衣可以選擇性地在胃腸道裡溶解。 Gelatin capsules comprising the compounds disclosed herein and/or at least one pharmaceutically acceptable salt and powdered carrier, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid and the like, may also be employed. A similar diluent can be used for tableting. Both tablets and capsules can be formulated as sustained release products for continued administration over a period of time. Compressed tablets may be coated with sugar or film to mask an unpleasant taste while protecting the tablet from air, or the enteric coating may be selectively dissolved in the gastrointestinal tract.
用於口服的液體劑型進一步包括至少一種選自於著色劑和矯味劑，用來提升患者的接受度。 Liquid dosage forms for oral administration further comprise at least one selected from the group consisting of coloring agents and flavoring agents for enhancing patient acceptance.
通常，水，合適的油，鹽溶液，葡萄糖水溶液(glucose)，及相關的糖溶液和二醇類物質比如丙二醇或者聚乙二醇都可以是非腸道給藥溶液的合適載體。非腸道給藥的溶液包含本發明所述的至少一個化合物的水溶性鹽，至少一個合適的穩定劑，及在必要情況下至少一種緩衝物。一些抗氧化劑比如亞硫酸氫鈉，亞硫酸鈉，或者抗壞血酸，單獨或者連用，都可以作為合適的穩定劑。檸檬酸及其鹽和乙二胺四乙酸鈉也可以作為合適的穩定劑。除此之外，非腸道給藥溶液可以進一步包含至少一種防腐劑，選自於，比如，苯紮氯胺，甲基-和丙基對羥基苯甲酸酯和氯丁醇。 In general, water, a suitable oil, a saline solution, a glucose solution, and related sugar solutions and glycols such as propylene glycol or polyethylene glycol may be suitable carriers for parenteral solutions. Solutions for parenteral administration comprise a water-soluble salt of at least one compound of the invention, at least one suitable stabilizer, and, if necessary, at least one buffer. Some antioxidants such as sodium bisulfite, sodium sulfite, or ascorbic acid, either alone or in combination, can be used as suitable stabilizers. Citric acid and its salts and sodium edetate can also be used as suitable stabilizers. In addition to this, the parenteral administration solution may further comprise at least one preservative selected from, for example, benzalkonium chloride, methyl- and propyl-p-hydroxybenzoate and chlorobutanol.
藥學上可以接受的載體可以選自於，比如能和藥物組合中的活性成分相容(在一些具體的實例中，可以穩定活性成分)和不會對受試者有害的載體。例如一些增溶劑環糊精(能夠和本發明公開的的至少一種化合物和/或者至少一種藥學上可接受的鹽形成特定的，更易溶的複合物)可以用作藥用輔料來傳輸活性成分。其它載體包括膠狀二氧化矽，硬脂酸鎂，纖維素，十二烷基硫酸鈉，和一些色素比如D&C黃色10號。一些合適的藥學上可接受的載體在Remington's Pharmaceutical Sciences,A.Osol中已有描述，可以用作本技術領域的標準文本。 The pharmaceutically acceptable carrier can be selected, for example, from a carrier which is compatible with the active ingredient in the pharmaceutical combination (in some specific examples, the active ingredient can be stabilized) and which is not deleterious to the subject. For example, some solubilizing cyclodextrins (which are capable of forming a specific, more soluble complex with at least one compound and/or at least one pharmaceutically acceptable salt disclosed herein) can be used as a pharmaceutical excipient to deliver the active ingredient. Other carriers include colloidal cerium oxide, magnesium stearate, cellulose, sodium lauryl sulfate, and some pigments such as D&C Yellow No. 10. Some suitable pharmaceutically acceptable carriers are described in Remington's Pharmaceutical Sciences , A. Osol and can be used as standard text in the art.
本發明中的化合物及其異構體，和藥學上可以接受的鹽可以進一步用於檢驗在體內對Btk相關疾病的療效。比如，本發明公開的化合物和/或者至少一種藥學上可接受的鹽可以對患有與Btk相關疾病的動物(比如小鼠模型)給藥，進而評價它的治療效 果。在一個或者多個這種試驗中得到陽性結果可以有效地增強科學知識寶庫，進而有效地顯示這些化合物和/或者鹽的實用性。基於這些結論，可以確定對動物和人的給藥劑量和給藥途徑。 The compounds of the present invention, and isomers thereof, and pharmaceutically acceptable salts can be further used to test the efficacy of Btk-related diseases in vivo. For example, a compound disclosed in the present invention and/or at least one pharmaceutically acceptable salt can be administered to an animal (such as a mouse model) having a disease associated with Btk, thereby evaluating its therapeutic effect. fruit. A positive result in one or more of these tests can effectively enhance the treasure trove of scientific knowledge, thereby effectively demonstrating the utility of these compounds and/or salts. Based on these conclusions, the dosage and route of administration to animals and humans can be determined.
對於吸入式給藥方式，本發明公開的化合物，及其異構體，和藥學上可以接受的鹽可以方便地從壓縮器或者噴霧器中以氣溶膠噴霧的形式傳輸。本發明公開的的化合物及其異構體，和藥學上可以接受的鹽也可以以一定劑型的粉末或者在噴射乾粉吸入器裝置輔助下吸入粉末組合物的形式傳輸。定量劑量吸入噴霧器(MDI)是典型的吸入式傳輸系統，可以將本發明公開的化合物及其異構體，和藥學上可以接受的鹽製成在至少一種推進物比如選自碳氟化合物和烴類化合物中的懸濁液或者溶液。 For inhaled administration, the compounds disclosed herein, and isomers thereof, and pharmaceutically acceptable salts can be conveniently delivered as an aerosol spray from a compressor or nebulizer. The compounds disclosed herein, and isomers thereof, and pharmaceutically acceptable salts can also be delivered in the form of a powder of a dosage form or inhalation of a powder composition with the aid of a spray dry powder inhaler device. A metered dose inhalation nebulizer (MDI) is a typical inhalation delivery system that can make the compounds disclosed herein, and isomers thereof, and pharmaceutically acceptable salts, in at least one propellant, such as selected from the group consisting of fluorocarbons and hydrocarbons. A suspension or solution in a compound.
對於眼部給藥，眼用製劑可以通過將適量重量百分比的本發明公開的化合物及其異構體，和藥學上可以接受的鹽的溶液或者懸浮液，製成一種合適的眼用載體，使得比如本發明公開的化合物，及其異構體，和藥學上可以接受的鹽與眼球表面保持足夠時間的接觸，從而能讓化合物滲透到眼睛的角膜和內部區域。 For ophthalmic administration, ophthalmic preparations can be formulated into a suitable ophthalmic vehicle by dissolving an appropriate amount by weight of a compound of the present invention, and isomers thereof, and a pharmaceutically acceptable salt. For example, the compounds disclosed herein, and their isomers, and pharmaceutically acceptable salts remain in contact with the surface of the eye for a sufficient period of time to allow penetration of the compound into the cornea and internal regions of the eye.
本發明公開的化合物，及其異構體，和藥學上可以接受的鹽有用的藥用劑型包括但不限於，硬和軟明膠膠囊，藥片，腸外注射劑，和口服懸浮液。 Pharmaceutically acceptable dosage forms of the presently disclosed compounds, and isomers thereof, and pharmaceutically acceptable salts include, but are not limited to, hard and soft gelatin capsules, tablets, parenteral injections, and oral suspensions.
服藥劑量和許多因素相關，比如年齡，受試者的健康程度和體重，疾病的程度，同時治療的種類，若有的話，治療的頻率和期望的效果。總的來說，活性成分的日用劑量是可以變化的，比 如，可以從每天0.1到2000毫克。例如，10-500毫克一天一次或者多次可能達到期望的效果。 The dose is related to a number of factors, such as age, the health and weight of the subject, the extent of the disease, and the type of treatment, if any, the frequency of treatment and the desired effect. In general, the daily dose of the active ingredient can vary, For example, it can be from 0.1 to 2000 mg per day. For example, 10-500 mg once or more times a day may achieve the desired effect.
在一些實施例中，大量的單位膠囊可以每個填充標準的兩片硬明膠膠囊製備，例如，比如100毫克粉末形式的本發明公開的化合物及其異構體，和藥學上可以接受的鹽，150毫克乳糖，50毫克纖維素，和6毫克硬脂酸鎂。 In some embodiments, a plurality of unit capsules can be prepared in two standard hard gelatin capsules filled with standard, such as, for example, 100 mg of the compound disclosed herein and isomers thereof, and a pharmaceutically acceptable salt, 150 mg lactose, 50 mg cellulose, and 6 mg magnesium stearate.
在一些實施例中，混合物選自於本發明中的化合物及其異構體，和藥學上可以接受的鹽和可消化的油，比如大豆油，棉花籽油或者橄欖油，可以通過主動取代泵入明膠的方式製備或者注入來形成包含100毫克活性成分的軟明膠膠囊。然後將膠囊洗滌和乾燥。 In some embodiments, the mixture is selected from the group consisting of the compounds of the invention and isomers thereof, and pharmaceutically acceptable salts and digestible oils, such as soybean oil, cottonseed oil or olive oil, which can be actively replaced by a pump The gelatin is prepared or infused to form a soft gelatin capsule containing 100 mg of the active ingredient. The capsules are then washed and dried.
在一些實施例中，大量的藥片可以通過常規程式製備，每個劑量單位包含，比如，100毫克選來自於本發明中的化合物及其異構體，和藥學上可以接受的鹽，0.2毫克膠體二氧化矽，5毫克硬脂酸鎂，275毫克微晶纖維素，11毫克澱粉和98.8毫克硬脂酸鎂。合適的包衣可以用於提高適口性或者延遲吸收。 In some embodiments, a plurality of tablets may be prepared by conventional procedures, each dosage unit comprising, for example, 100 mg of a compound derived from the present invention and an isomer thereof, and a pharmaceutically acceptable salt, 0.2 mg colloid. Ceria, 5 mg magnesium stearate, 275 mg microcrystalline cellulose, 11 mg starch and 98.8 mg magnesium stearate. A suitable coating can be used to increase palatability or delay absorption.
在一些實施例中，適用於注射給藥的胃腸外組合物，可以通過攪拌1.5%重量比的本發明公開的化合物和/或至少一個對映體，非對映異構體，或者它們藥學上可接受的鹽，在10%體積比的丙二醇中製備。該溶液加入用注射用水到預期體積，再滅菌。 In some embodiments, a parenteral composition suitable for administration by injection may be stirred by stirring 1.5% by weight of a compound of the present invention and/or at least one enantiomer, diastereomer, or pharmaceutically thereof. An acceptable salt is prepared in 10% by volume of propylene glycol. The solution is added to the intended volume with water for injection and sterilized.
在一些實施例中，可製備用於口服給藥的水溶性懸浮液。例如，每一個5毫升的水懸浮液中含有100毫克細碎的一種 化合物，其立體異構體，及其藥學上可接受的鹽，100毫克羧甲基纖維素鈉，5毫克苯甲酸鈉，1克山梨醇溶液，U.S.P.和0.025毫升香草醛。 In some embodiments, a water soluble suspension for oral administration can be prepared. For example, each 5 ml of aqueous suspension contains 100 mg of a finely divided one. A compound, a stereoisomer thereof, and a pharmaceutically acceptable salt thereof, 100 mg of sodium carboxymethylcellulose, 5 mg of sodium benzoate, 1 g of sorbitol solution, U.S.P. and 0.025 ml of vanillin.
當本發明中的化合物，及其異構體，和藥學上可以接受的鹽與至少一種其它的治療試劑逐步或者一起給藥時，可以使用同樣的劑型。當藥物在物理組合給藥時，劑型和給藥途徑的選擇取決於組合藥物的相容性。因此這一術語同時給藥可以理解為包含至少兩種試劑同時或者依次給藥，或者以固定劑量的至少兩種活性成分的組合物給藥。 The same dosage form can be used when the compounds of the invention, and isomers thereof, and pharmaceutically acceptable salts are administered stepwise or together with at least one other therapeutic agent. When the drug is administered in a physical combination, the choice of dosage form and route of administration will depend on the compatibility of the combination drug. Thus, the simultaneous administration of the term is understood to mean the administration of at least two agents simultaneously or sequentially, or in a fixed dose of a combination of at least two active ingredients.
本發明公開的化合物，及其異構體，和藥學上可以接受的鹽可以以單一活性成分給藥或者和至少一種第二種活性成分，比如，選自公知的用於治療Btk相關疾病的其他活性成分聯合給藥。 The compounds disclosed herein, and the isomers thereof, and the pharmaceutically acceptable salts can be administered as a single active ingredient or together with at least one second active ingredient, for example, selected from other known compounds for the treatment of Btk-related diseases. The active ingredients are administered in combination.
可以理解的是，本文所述的實施例和實施方案僅僅是出於列舉說明的目的，本領域技術人員可以據此進行各種修飾或者改變，並且這些修飾和改變均包括在本發明的主旨和範圍內以及所附申請專利範圍的保護範圍之內。本文中所引證的所有公開出版物、專利和專利申請的全部內容均引入本文作為參考，其用於所有目的。 It is to be understood that the embodiments and the embodiments described herein are for the purpose of illustration only, and that various modifications and changes can be made by those skilled in the art, and such modifications and changes are included in the spirit and scope of the present invention. It is within the scope of protection of the scope of the appended claims. All publications, patents and patent applications cited herein are hereby incorporated by reference in their entirety in their entirety in their entirety herein
化合物合成的常規方案 Conventional scheme for compound synthesis
本發明中的化合物以及它們在藥學上可接受的鹽的製備可以通過(a)市售的起始原料(b)文獻中記載的已知起始原料(c) 在此合成方案及實驗步驟中描述的中間體。在合成本發明中的這些化合物時，可能需要變化合成步驟的先後順序來提高產率。本發明中的一些化合物可以通過以下合成方案和描述來合成。 The compounds of the present invention and their pharmaceutically acceptable salts can be prepared by (a) a commercially available starting material (b) known starting materials (c) as described in the literature. The intermediates described in this synthetic scheme and experimental procedure. In synthesizing these compounds of the invention, it may be necessary to vary the order of the synthetic steps to increase the yield. Some of the compounds of the present invention can be synthesized by the following synthetic schemes and descriptions.
方案I給出了本發明中化合物S-6的常規合成路線，其中A,R1,R2,R4,R6,L和n如本文所述。甲基酮S-1與N,N-二烷基甲醯胺的縮醛或者N,N-二烷基乙醯胺的縮醛在回流溫度下反應幾個小時，得到3-二烷基氨基-1-(芳基，雜芳基或烷基)-2-丙烯-1-酮S-2。製備中間體S-3的方法基本上與國際專利公開號為WO2001/019829和WO2011/046964中描述的類似。中間體S-3和帶有取代的3-二烷基氨基-1-(芳基，雜芳基或烷基)-2-丙烯-1-酮S-2在弱酸如乙酸，或在含有催化量酸的惰性溶劑中如甲苯，乙腈或二甲氧基乙烷，80℃或者回流溫度下反應幾個小時，得到腈化合物S-4。用還原劑如硼氫化鈉(NaBH4)或NaBH4隨後用Pd/C還原嘧啶環得到四氫吡唑並嘧啶S-5。隨後，在醇溶液中鹼性條件 下例如NaOH或KOH加H2O2，或者酸性條件下例如H3PO4,H2SO4，或者BF3．HOAc水解氰基得到甲醯胺S-6。 Scheme I gives a general synthetic route for the compound S-6 of the present invention wherein A, R 1 , R 2 , R 4 , R 6 , L and n are as described herein. S-1 methyl ketone and N, N - dialkyl acetal A Amides or N, N - dialkyl acetal acetyl amine at reflux temperature for several hours, to give 3-amino-alkyl 1-(Aryl, heteroaryl or alkyl)-2-propen-1-one S-2 . The process for the preparation of the intermediate S-3 is substantially similar to that described in the international patent publications WO2001/019829 and WO2011/046964. Intermediate S-3 and substituted 3-dialkylamino-1-(aryl,heteroaryl or alkyl)-2-propen-1-one S-2 in a weak acid such as acetic acid or in the presence of a catalyst The acid-containing inert solvent such as toluene, acetonitrile or dimethoxyethane is reacted at 80 ° C or reflux temperature for several hours to obtain a nitrile compound S-4 . Reduction of the pyrimidine ring with a reducing agent such as sodium borohydride (NaBH 4 ) or NaBH 4 followed by Pd/C affords the tetrahydropyrazolopyrimidine S-5 . Subsequently, under alkaline conditions such as NaOH or KOH plus H 2 O 2 in an alcoholic solution, or under acidic conditions such as H 3 PO 4 , H 2 SO 4 , or BF 3 . Hydrolysis of the cyano group by HOAc gives the methotrexate S-6 .
方案II給出了本發明中甲醯胺S-13的常規合成路線，其中A,R1,R2,R4,R5,L,n和p如本文所述。保護的肼S-7可以方便地通過文獻中描述的方法(J.Med.Chem.1992,35,2392)製備。用酸脫保護，隨後這個中間體與中間體S-10(國際專利公開號為WO2001/019829和WO2011/046964中有描述)，在鹼性溶劑如TEA/乙醇，縮合得到吡唑酯S-9。吡唑醇S-11的製備可以通過還原酯類化合物S-9。可被用於此還原過程中的還原劑包括，但不限於LiBH4，NaBH4和超氫化物。分子內N-烷基化或還原胺化，得到腈類化合物S-12，通過方案I中描述的相同方法將其轉化成甲醯胺S-13。 Scheme II gives a conventional synthetic route for the formamide S-13 of the present invention, wherein A, R 1 , R 2 , R 4 , R 5 , L, n and p are as described herein. The protected oxime S-7 can be conveniently prepared by the method described in the literature (J. Med. Chem. 1992, 35, 2392). Deprotection with acid, followed by condensation of the intermediate with intermediate S-10 (described in International Patent Publication Nos. WO2001/019829 and WO2011/046964) in a basic solvent such as TEA/ethanol to give the pyrazole ester S-9 . The pyrazole alcohol S-11 can be prepared by reducing the ester compound S-9 . Reducing agents that can be used in this reduction process include, but are not limited to, LiBH 4 , NaBH 4 and super hydride. Intramolecular N -alkylation or reductive amination affords the nitrile compound S-12 which is converted to the formamide S-13 by the same procedure as described in Scheme I.
方案IIIOption III
方案III給出了本發明中合成甲醯胺S-13的替代路線。 Scheme III gives an alternative route to the synthesis of methotrexate S-13 in the present invention.
方案IV給出了本發明中S-19和S-20的合成路線，其中A,R1,R2,R4,R5,L和n如本文所述。使用方案I中相同的方 法水解腈類化合物S-3得到甲醯胺S-18。吡唑甲醯胺S-18或者吡唑腈類化合物S-3與市售或者製備的鹵代酮或者鹵代醛S-23發生環化反應得到位置異構體S-19和S-20，或者S-21和S-22。腈類化合物S-21和S-22根據方案I水解得到甲醯胺S-19和S-20。 Scheme IV gives the synthetic route for S-19 and S-20 in the present invention, wherein A, R 1 , R 2 , R 4 , R 5 , L and n are as described herein. The nitrile compound S-3 was hydrolyzed in the same manner as in Scheme I to give the formamide S-18 . The pyrazole meglumine S-18 or the pyrazole nitrile compound S-3 is cyclized with a commercially available or prepared halo ketone or a haloaldehyde S-23 to give positional isomers S-19 and S-20 , Or S-21 and S-22 . The nitrile compounds S-21 and S-22 were hydrolyzed according to Scheme I to give the carbamides S-19 and S-20 .
方案V給出了本發明中S-27的合成路線，其中A,R1,R2,R4,R6,L,m和n如本文所述。吡唑S-25的製備通過中間體S-10與市售或製備的肼S-24在醇中環化反應所得，可以參考方案Ⅱ中所述的方法。中間體S-25進行分子內環化，包括但並不限於Cu催化偶聯，Buchwald反應，SN2反應或還原胺化反應。最終，腈類化合物S-26根據方案I水解得到甲醯胺S-27。或者，親核取代反應，然後通過分子內環化，得到甲醯胺S-27。 Scheme V gives a synthetic route for S-27 in the present invention, wherein A, R 1 , R 2 , R 4 , R 6 , L, m and n are as described herein. The preparation of pyrazole S-25 is carried out by cyclization of the intermediate S-10 with commercially available or prepared hydrazine S-24 in an alcohol, and the method described in Scheme II can be referred to. Intermediate S-25 undergoes intramolecular cyclization including, but not limited to, Cu catalyzed coupling, Buchwald reaction, SN2 reaction or reductive amination. Finally, the nitrile compound S-26 was hydrolyzed according to Scheme I to give the formamide S-27 . Alternatively, the nucleophilic substitution reaction is followed by intramolecular cyclization to give the methotrexate S-27 .
方案VIProgram VI
方案VI給出了本發明中S-30的合成路線，其中A,R1,R2,R4,R6,L,m和n如本文所述。分子內醯胺S-30的製備通過腈類化合物S-3或甲醯胺S-18和酯S-29在惰性溶劑如DMF，堿如K2CO3和TEA存在下加熱得到。 Scheme VI gives the synthetic route for S-30 in the present invention, wherein A, R 1 , R 2 , R 4 , R 6 , L, m and n are as described herein. The preparation of the in - molecular indoleamine S-30 is obtained by heating the nitrile compound S-3 or the formamide S-18 and the ester S-29 in the presence of an inert solvent such as DMF, such as K 2 CO 3 and TEA.
方案VII給出了本發明中S-32的合成路線，其中A,R1,R2,R4,L和n如本文所述。醯胺S-32的製備通過腈類化合物S-3或甲醯胺S-18和β-酮酯S-31在弱酸如乙酸，或在含有催化量酸的惰性溶劑如甲苯，乙腈或二甲氧基乙烷加熱得到。 Scheme VII gives a synthetic route for S-32 in the present invention wherein A, R 1 , R 2 , R 4 , L and n are as described herein. The indoleamine S-32 is prepared by the nitrile compound S-3 or the carbamide S-18 and the β-ketoester S-31 in a weak acid such as acetic acid, or in an inert solvent containing a catalytic amount of acid such as toluene, acetonitrile or dimethyl The oxyethane is heated to obtain.
方案VIIIOption VIII
方案VIII給出了本發明中S-36的合成路線，其中A,R1,R2,R6,R7,L,W,m和n如本文所述。腈類化合物S-3或甲醯胺S-18和市售或製備的中間體S-33在惰性溶劑如DMF中，鹼性如K2CO3和TEA條件下，發生分子間環化反應得到化合物S-36。或者，腈類化合物S-3或甲醯胺S-18和二醛類化合物S-34縮合，然後還原得到化合物S-36。 Scheme VIII gives a synthetic route for S-36 in the present invention wherein A, R 1 , R 2 , R 6 , R 7 , L, W, m and n are as described herein. Intermolecular cyclization of the nitrile compound S-3 or formamide S-18 and the commercially available or prepared intermediate S-33 in an inert solvent such as DMF under basic conditions such as K 2 CO 3 and TEA Compound S-36 . Alternatively, the nitrile compound S-3 or the meglumine S-18 and the dialdehyde compound S-34 are condensed and then reduced to give the compound S-36 .
實施例Example
下面的實施例純粹是示例性的，不應被認為以任何方式限制。已作出努力確保使用資料的準確性(比如，數量，溫度等)，但仍然會有一些實驗方面的誤差和偏差。除非另有說明，溫度是指攝氏度。試劑購自Sigma-Aldrich，Alfa Aesar，或者TCI等供應商，除非另有說明，使用時未作進一步的純化。 The following examples are purely exemplary and should not be considered limiting in any way. Efforts have been made to ensure the accuracy of the data used (eg, quantity, temperature, etc.), but there will still be some experimental errors and deviations. Unless otherwise stated, temperature refers to degrees Celsius. Reagents were purchased from suppliers such as Sigma-Aldrich, Alfa Aesar, or TCI and were not further purified for use unless otherwise indicated.
除非另有說明，如下反應一般在氮氣或者氬氣的正壓下或者帶有乾燥管的無水溶劑中進行；反應瓶帶有橡膠塞以便於通過注射器加入底物和試劑；玻璃器皿通過烘乾和/或者加熱乾燥。 1H NMR使用400MHz的Agilent儀器得到。氫譜採用CDCl3,CD2Cl2,CD3OD,D2O,d 6-DMSO,d 6-丙酮or(CD3)2CO作溶劑，四甲基矽(0.00ppm)或者溶劑殘餘(CDCl3：7.25ppm；CD3OD：3.31ppm；D2O：4.79ppm；d 6-DMSO：2.50ppm；d 6-丙酮：2.05；(CD3)2CO：2.05)為參考標準。當峰的多重性報導時，可以使用如下的簡寫：s(單峰)，d(雙峰)，t(三重峰)，q(四重峰)，qn(五重峰)，sx(六重峰)，m(多重峰)，br(寬峰)，dd(兩個雙峰)，dt(兩三重峰)。給出的耦合常數用赫茲(Hz)表示。 Unless otherwise stated, the following reactions are generally carried out under a positive pressure of nitrogen or argon or in an anhydrous solvent with a drying tube; the reaction flask is provided with a rubber stopper to facilitate the addition of the substrate and reagents via a syringe; the glassware is dried and / or heat dry. 1 H NMR was obtained using a 400 MHz Agilent instrument. Hydrogen spectrum using CDCl 3 , CD 2 Cl 2 , CD 3 OD, D 2 O, d 6 -DMSO, d 6 -acetone or (CD 3 ) 2 CO as solvent, tetramethylguanidine (0.00 ppm) or solvent residue ( CDCl 3 : 7.25 ppm; CD 3 OD: 3.31 ppm; D 2 O: 4.79 ppm; d 6 -DMSO: 2.50 ppm; d 6 -acetone: 2.05; (CD 3 ) 2 CO: 2.05) is a reference standard. When the multiplicity of peaks is reported, the following abbreviations can be used: s (single peak), d (double peak), t (triplet), q (quadruple), qn (five peak), sx (six weights) Peak), m (multiple peak), br (wide peak), dd (two double peaks), dt (two or three heavy peaks). The coupling constants given are expressed in hertz (Hz).
高效液相色譜-質譜聯用光譜儀(Agilent 1260)檢測器：MWD(190-400nm)，質譜檢測器：6120 SQ High Performance Liquid Chromatography-Mass Spectrometry (Agilent 1260) Detector: MWD (190-400 nm), Mass Spectrometer Detector: 6120 SQ
流動相：A：含有0.1%甲酸的乙腈，B：含有0.1%甲酸的水 Mobile phase: A: acetonitrile with 0.1% formic acid, B: water with 0.1% formic acid
反相柱：Poroshell 120 EC-C18,4.6×50mm,2.7μm Reversed phase column: Poroshell 120 EC-C18, 4.6×50mm, 2.7μm
梯度方法：流速：1.8mL/min Gradient method: Flow rate: 1.8 mL/min
製備高效液相色譜執行條件：反相柱(150×21.2mm的ID，5μm，Gemini NX-C18)，流速為20mL/min，進樣量2mL，室溫，在214nm和254nm進行紫外檢測。 Preparation of high performance liquid chromatography Execution conditions: reverse phase column (150 × 21.2 mm ID, 5 μm, Gemini NX-C18), flow rate of 20 mL / min, injection volume 2 mL, room temperature, UV detection at 214 nm and 254 nm.
在下面的實施例中，使用到下面的縮寫：
實施例1：化合物1-4的合成Example 1: Synthesis of Compound 1-4
化合物1：7-(3-氨基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈 Compound 1 : 7-(3-Aminophenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile
步驟1：2-(羥基(4-苯氧基苯基)亞甲基)丙二腈 Step 1: 2-(hydroxy(4-phenoxyphenyl)methylene)malononitrile
向4-苯氧基苯甲酸(300g,1.4mol)中加入SOCl2(1.2L)，加熱至80℃，攪拌3小時。混合物減壓濃縮得到中間體(315g)，未經進一步純化，直接用於下一步反應。 To 4-phenoxybenzoic acid (300 g, 1.4 mol) was added SOCl 2 (1.2 L), heated to 80 ° C, and stirred for 3 hours. The mixture was concentrated under reduced pressure to give EtOAc m.
在0~5℃，向含有丙二腈(89.5g,1355mmol)和DIEA(350g,2710mmol)的THF(800mL)溶液中滴加中間體(315g)的甲苯(800mL)溶液，約2小時。混合物允許恢復到室溫，攪拌16小時。反應用水(2.0L)淬滅，EA(2.0L×3)萃取。合併有機相，依次用稀鹽酸(3N,1L)，飽和食鹽水(2.0L×3)洗滌，Na2SO4乾燥，濃縮，得到黃色固體(330g,93%)。1H NMR(DMSO-d6)δ 7.62(d,J=8.8Hz,2H),7.46-7.38(m,2H),7.18(t,J=7.6Hz,1H),7.06(d,J=8.0Hz,2H),6.94(d,J=8.8Hz,2H).MS(ESI)m/e[M+1]+ 262.9。 To a solution of malononitrile (89.5 g, 1355 mmol) and DIEA (350 g, 2710 mmol) in THF (800 mL The mixture was allowed to return to room temperature and stirred for 16 hours. The reaction was quenched with water (2.0 L) and EtOAc (EtOAc. The combined organic phase was washed with dilute hydrochloric acid (3N, 1L), washed with saturated brine and dried (2.0L × 3) Na 2 SO 4, and concentrated to give a yellow solid (330g, 93%). 1 H NMR (DMSO-d 6 ) δ 7.62 (d, J = 8.8 Hz, 2H), 7.46-7.38 (m, 2H), 7.18 (t, J = 7.6 Hz, 1H), 7.06 (d, J = 8.0 Hz, 2H), 6.94 (d, J = 8.8 Hz, 2H). MS (ESI) m/e [M+1] + 262.9.
步驟2：2-(甲氧基(4-苯氧基苯基)亞甲基)丙二腈 Step 2: 2-(Methoxy(4-phenoxyphenyl)methylene)malononitrile
向原甲酸三甲酯(500mL)中加入2-(羥基(4-苯氧基苯基)亞甲基)丙二腈(50g,190.8mmol)，加熱至75℃，攪拌16小時。混合物濃縮，殘餘物用MeOH(50mL)洗滌，得到黃色固體產品2-(甲氧基(4-苯氧基苯基)亞甲基)丙二腈(25g，47.5%)。1H NMR(DMSO-d6)δ 7.70(d,J=8.4Hz,2H),7.52-7.45(m,2H),7.28(t,J=7.6Hz,1H),7.22-7.06(m,4H),3.93(s,3H).MS(ESI)m/e[M+1]+ 276.9。 To a solution of trimethyl orthoformate (500 mL) was added 2-(hydroxy(4-phenoxyphenyl)methylene)malononitrile (50 g, 190.8 mmol), heated to 75 ° C and stirred for 16 hours. The mixture was concentrated and EtOAc EtOAc (EtOAc) 1 H NMR (DMSO-d 6 ) δ 7.70 (d, J = 8.4 Hz, 2H), 7.52-7.45 (m, 2H), 7.28 (t, J = 7.6 Hz, 1H), 7.22-7.06 (m, 4H) ), 3.93 (s, 3H). MS (ESI) m/e [M+1] + 276.9.
步驟3：5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈 Step 3: 5-Amino-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitrile
向2-(甲氧基(4-苯氧基苯基)亞甲基)丙二腈(80g,290mmol)的乙醇(200mL)溶液中加入水合肼(20mL)，混合物室溫攪拌16小時。混合物濃縮得到粗品，用MeOH(30mL)洗滌，得到類白色固體5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈(55g，68.8%)。1H NMR(DMSO-d6)δ 12.11(br s,1H),7.80(d,J=8.8Hz,2H),7.46-7.39(m,2H),7.18(t,J=7.6Hz,1H),7.12-7.04(m,4H),6.43(br s,2H)。 To a solution of 2-(methoxy(4-phenoxyphenyl)methylene)malononitrile (80 g, 290 mmol) in EtOAc (20 mL) The mixture was concentrated to give a crystal crystal crystal crystal crystal crystal crystal crystal crystal crystal crystal crystal crystal crystal crystal crystal crystal crystal crystal crystal crystal crystal crystal crystal 1 H NMR (DMSO-d 6 ) δ 12.11 (br s, 1H), 7.80 (d, J = 8.8 Hz, 2H), 7.46-7.39 (m, 2H), 7.18 (t, J = 7.6 Hz, 1H) , 7.12-7.04 (m, 4H), 6.43 (br s, 2H).
步驟4：(E)-N-(3-(3-(二甲基氨基)丙烯基醯基)苯基)乙醯胺 Step 4: (E) -N- (3-(3-(Dimethylamino)propenyl)phenyl)acetamide
氮氣下，將N-(3-乙醯基苯基)乙醯胺(1.77g,10.0mmol)溶於DMF-DMA(6mL)，加入分子篩(10粒)，100℃下攪拌2小時。濃縮混合物，並用甲基叔丁基醚(30mL)洗滌，得到黃色固體(E)-N-(3-(3-(二甲基氨基)丙烯基醯基)苯基)乙醯胺(2.1g,90%)。 N-(3-Ethylphenyl)acetamide (1.77 g, 10.0 mmol) was dissolved in DMF-DMA (6 mL) under nitrogen, and molecular sieves (10) were added and stirred at 100 ° C for 2 hours. The mixture was concentrated, and washed with methyl tert-butyl ether (30 mL), to give a yellow solid (E) - N - (3- (3- ( dimethylamino) acyl-propenyl) phenyl) acetyl amine (2.1g , 90%).
步驟5：N-(3-(3-氰基-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-7-基)苯基)乙醯胺 Step 5: N- (3-(3-Cyano-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidin-7-yl)phenyl)acetamide
向(E)-N-(3-(3-(二甲基氨基)丙烯基醯基)苯基)乙醯胺(46mg,0.2mmol)的醋酸(5mL)溶液中，加入5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈(55mg,0.2mmol)。混合物在118℃攪拌4小時。將反應混合物濃縮，殘餘物加入到乙酸乙酯(100mL)和飽和食鹽水(100mL)中。有機相用飽和食鹽水(100mL×2)洗滌，硫酸鈉乾燥並濃縮，濃縮得到無色油狀物N-(3-(3-氰基-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-7-基)苯基)乙醯胺(80mg,90%)。MS(ESI)m/e[M+1]+ 446。 To a solution of (E) -N- (3-(3-(dimethylamino)propenyl) phenyl)acetamide (46 mg, 0.2 mmol) in EtOAc (5 mL) -(4-Phenoxyphenyl)-1H-pyrazole-4-carbonitrile (55 mg, 0.2 mmol). The mixture was stirred at 118 ° C for 4 hours. The reaction mixture was concentrated and the residue was crystalljjjjjjjjjj The organic phase was washed with saturated brine (100mL × 2), dried over sodium sulfate and concentrated to give concentrated to a colorless oil N - (3- (3- cyano-2- (4-phenoxyphenyl) pyrazolo [1,5-a]pyrimidin-7-yl)phenyl)acetamide (80 mg, 90%). MS (ESI) m / e [M + 1] + 446.
步驟6：7-(3-氨基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈 Step 6: 7-(3-Aminophenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile
向N-(3-(3-氰基-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-7-基)苯基)乙醯胺(285mg,0.64mmol)的乙醇(6mL)溶液中，加入鹽酸(3mL)。混合物在75℃攪拌3小時。濃縮，得到黃色固體7-(3-氨基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈(250mg,97%)。1H NMR(400MHz,DMSO-d6)δ 8.90(d,J=4.4Hz,1H),8.67(br s,2H),8.15(d,J=8.8Hz,2H),8.06(s,1H),7.87(d,J=8.0Hz,1H),7.64(t,J=8.0Hz,1H),7.57(d,J=4.4Hz,1H),7.48-7.44(m,3H),7.25-7.18(m,3H),7.13(d,J=8.0Hz,2H).MS(ESI)m/e[M+1]+ 404。 To N- (3-(3-cyano-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidin-7-yl)phenyl)acetamidamine (285 mg, 0.64 mmol) To a solution of ethanol (6 mL), hydrochloric acid (3 mL) was added. The mixture was stirred at 75 ° C for 3 hours. Concentration gave 7-(3-aminophenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile (250 mg, 97%). 1 H NMR (400MHz, DMSO- d 6) δ 8.90 (d, J = 4.4Hz, 1H), 8.67 (br s, 2H), 8.15 (d, J = 8.8Hz, 2H), 8.06 (s, 1H) , 7.87 (d, J = 8.0 Hz, 1H), 7.64 (t, J = 8.0 Hz, 1H), 7.57 (d, J = 4.4 Hz, 1H), 7.48-7.44 (m, 3H), 7.25-7.18 ( m, 3H), 7.13 (d, J = 8.0 Hz, 2H). MS (ESI) m/e [M+1] + 404.
化合物2：7-(3-氨基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 2 : 7-(3-Aminophenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-A Guanamine
步驟1：7-(3-氨基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈 Step 1: 7-(3-Aminophenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-A Nitrile
將硼氫化鈉(70mg,1.86mmol)加入到7-(3-氨基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈(150mg,0.372mmol)的乙醇(10mL)溶液中。混合物室溫攪拌16小時，60℃攪拌2小時。然後將反應混合物濃縮，殘餘物加入到乙酸乙酯(50mL)和飽和食鹽水(40mL)中。從水層中分離出有機層，飽和食鹽水(50mL×2)洗滌，Na2SO4乾燥，濃縮得到黃色固體7-(3-氨基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈(135mg，粗品)。MS(ESI)m/e[M+1]+ 408。 Sodium borohydride (70 mg, 1.86 mmol) was added to 7-(3-aminophenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile ( 150 mg, 0.372 mmol) in ethanol (10 mL). The mixture was stirred at room temperature for 16 hours and at 60 ° C for 2 hours. The reaction mixture was concentrated and the residue was crystalljjjjjjjjjj The organic layer was separated from the aqueous layer, washed with saturated brine (50 mL×2), dried over Na 2 SO 4 and concentrated to give a yellow solid 7-(3-aminophenyl)-2-(4-phenoxyphenyl) -4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carbonitrile (135 mg, crude). MS (ESI) m / e [M + 1] + 408.
步驟2：7-(3-氨基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 2: 7-(3-Aminophenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-A Guanamine
將5N的NaOH水溶液(1mL)和H2O2(1mL)加入到7-(3-氨基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈(130mg,0.32mmol)的DMSO(2mL)和乙醇(2mL)溶液中。將混合物在60℃攪拌30分鐘，濃縮，殘餘物加入到乙酸乙酯(100mL)和飽和食鹽水(100mL)中。分離出有機層，飽和食鹽水洗滌(100mL×3)，Na2SO4乾燥，矽膠層析柱(石油醚/乙酸乙酯洗脫)純化，得到黃色固體7-(3-氨基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺(35mg,26%)。1H NMR(400MHz,DMSO-d6)δ 7.50(d,J=8.4Hz,2H),7.40-7.33(m,2H),7.16(t,J=7.6Hz,1H),7.06(d,J=8.0Hz,2H),7.03(d,J=8.4Hz,2H),6.97(t,J=7.6Hz,1H),6.76(s,1H),6.44(d,J=8.4Hz,1H),6.23-6.21(m,2H),5.30-5.25(m,1H),5.09(s,2H),3.30-3.28(m,1H),3.12-3.02(m,1H),2.34-2.26(m,1H),2.05-2.01(m,1H).MS(ESI)m/e[M+1]+ 426。 Add 5N aqueous NaOH (1 mL) and H 2 O 2 (1 mL) to 7-(3-aminophenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydrogen A solution of pyrazolo[1,5-a]pyrimidine-3-carbonitrile (130 mg, 0.32 mmol) in DMSO (2 mL) and ethanol (2 mL). The mixture was stirred at 60 <0>C for 30 min, concentrated and EtOAc EtOAc (EtOAc) The organic layer was separated, washed with saturated brine (100 mL×3), dried over Na 2 SO 4 2-(4-Phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide (35 mg, 26%). 1 H NMR (400MHz, DMSO- d 6) δ 7.50 (d, J = 8.4Hz, 2H), 7.40-7.33 (m, 2H), 7.16 (t, J = 7.6Hz, 1H), 7.06 (d, J = 8.0 Hz, 2H), 7.03 (d, J = 8.4 Hz, 2H), 6.97 (t, J = 7.6 Hz, 1H), 6.76 (s, 1H), 6.44 (d, J = 8.4 Hz, 1H), 6.23-6.21(m,2H), 5.30-5.25(m,1H),5.09(s,2H), 3.30-3.28(m,1H),3.12-3.02(m,1H),2.34-2.26(m,1H) ), 2.05-2.01 (m, 1H). MS (ESI) m/e [M+1] + 426.
通過手性製備高效液相色譜法，化合物2被分離成兩個對映立體異構體2a(峰1，R或S，在手性分析的保留時間為8.94min)，和2b(峰2，S或R，在手性分析的保留時間為10.11min)。手性分離條件如下所示：
手性分析條件如下所示：
化合物3和4：7-(3-丙烯醯胺基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-α]嘧啶-3-甲醯胺和7-(3-(3-氯代丙醯胺基)苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-α]嘧啶-3-甲醯胺 Compounds 3 and 4 : 7-(3-Acrylaminophenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-α] Pyrimidine-3-carbamide and 7-(3-(3-chloropropionamido)phenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyridyl Oxazo[1,5-α]pyrimidine-3-carboxamide
向7-(3-氨基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺(30mg,0.071mmol)的DCM(2mL)溶液中，加入吡啶(0.2mL)。然後滴加丙烯醯氯(6mg,0.084mmol)。混合物室溫攪拌0.5小時，加入到二氯甲烷(20mL)和飽和食鹽水(20mL)中。從水層中分離出有機層，飽和食鹽水(2×20mL)洗滌，Na2SO4乾燥，並用製備矽膠板(DCM/CH3OH=10/1)純化，得到白色固體7-(3-丙烯醯胺基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-α]嘧啶-3-甲醯胺(1.82mg,5.38%)。1H NMR(400MHz,CD3OD)δ 7.58(d,J=8.8Hz,1H),7.46(d,J=8.8Hz,2H),7.34-7.26(m,4H),7.10(t,J=7.6Hz,1H),7.04-6.95(m,4H),6.84(d,J=7.6Hz,1H),6.39-6.25(m,2H),5.69(dd,J=2.4,9.6Hz,1H),5.47-5.44(m,1H),3.38-3.31(m,1H),3.22-3.12(m,1H),2.52-2.42(m,1H),2.23-2.17(m,1H).MS(ESI)m/e[M+1]+ 480。 To 7-(3-Aminophenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide (30 mg, 0.071 mmol) in EtOAc (2 mL). Propylene hydrazine chloride (6 mg, 0.084 mmol) was then added dropwise. The mixture was stirred at room temperature for 0.5 hr and added to dichloromethane (20 mL) and brine (20 mL). The organic layer was separated from the aqueous layer, washed with brine (2×20 mL), dried over Na 2 SO 4 and purified by silica gel (DCM/CH 3 OH = 10/1) to give a white solid 7-(3- Acrylamide phenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidine-3-carboxamide (1.82 mg , 5.38%). 1 H NMR (400 MHz, CD 3 OD) δ 7.58 (d, J = 8.8 Hz, 1H), 7.46 (d, J = 8.8 Hz, 2H), 7.34 - 7.26 (m, 4H), 7.10 (t, J = 7.6 Hz, 1H), 7.04-6.95 (m, 4H), 6.84 (d, J = 7.6 Hz, 1H), 6.39-6.25 (m, 2H), 5.69 (dd, J = 2.4, 9.6 Hz, 1H), 5.47-5.44 (m, 1H), 3.38-3.31 (m, 1H), 3.22-3.12 (m, 1H), 2.52-2.42 (m, 1H), 2.23-2.17 (m, 1H). MS (ESI) m /e[M+1] + 480.
白色固體副產物7-(3-(3-氯代丙醯胺基)苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-α]嘧啶-3-甲醯胺(2.21mg)。1H NMR(400MHz,CD3OD)δ 7.57(d,J=8.0Hz,1H),7.52(d,J=8.8Hz,2H),7.42-7.30(m,4H),7.16(t,J=7.6Hz,1H),7.09-7.01(m,4H),6.89(d,J=8.0Hz,1H),5.53-5.48(m,1H), 3.85(t,J=6.4Hz,2H),3.44-3.37(m,1H),3.26-3.19(m,1H),2.83(t,J=6.4Hz,2H),2.60-2.44(m,1H),2.33-2.22(m,1H).MS(ESI)m/e[M+1]+ 516.2。 White solid by-product 7-(3-(3-chloropropionamido)phenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1 , 5-α]pyrimidine-3-carboxamide (2.21 mg). 1 H NMR (400MHz, CD 3 OD) δ 7.57 (d, J = 8.0Hz, 1H), 7.52 (d, J = 8.8Hz, 2H), 7.42-7.30 (m, 4H), 7.16 (t, J = 7.6 Hz, 1H), 7.09-7.01 (m, 4H), 6.89 (d, J = 8.0 Hz, 1H), 5.53-5.48 (m, 1H), 3.85 (t, J = 6.4 Hz, 2H), 3.44 3.37(m,1H), 3.26-3.19(m,1H), 2.83(t, J =6.4Hz, 2H), 2.60-2.44(m,1H),2.33-2.22(m,1H).MS(ESI) m/e [M+1] + 516.2.
由2a和2b通過製備化合物3的類似方法，分別得到化合物3a(峰1，R或S，在手性分析的保留時間為4.45min)和3b(峰2，S或R，在手性分析的保留時間為7.41min)。 A similar method for the preparation of compound 3 from 2a and 2b gave compound 3a (peak 1, R or S, retention time in chiral analysis 4.45 min) and 3b (peak 2, S or R, in chiral analysis). The retention time is 7.41 min).
手性分析條件如下所示：
化合物5：7-(2-氨基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 5 : 7-(2-Aminophenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-A Guanamine
根據7-(3-氨基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈(步驟4到5)，化合物2(步驟1和2)和化合物68(步 驟8)的製備方案，在本領域普通技術人員熟知的條件下，以鄰硝基苯乙酮和5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈為起始原料，得到目標產物。1H NMR(DMSO-d6)δ 7.48-7.44(m,2H),7.40-7.33(m,2H),7.15-7.09(m,1H),7.05-6.97(m,4H),6.92(td,J=8.0,1.2Hz,1H),6.75(br s,1H),6.64(dd,J=8.0,1.2Hz,1H),6.46(td,J=7.6,1.2Hz,1H),6.23(dd,J=7.6,1.2Hz,1H),5.57-5.52(m,1H),5.19(br s,2H),3.27-3.17(m,1H),2.93(td,J=2.8,12.0Hz,1H),2.21-2.11(m,1H),2.10-2.05(m,1H).MS(ESI)m/e[M+1]+ 426.0。 According to 7-(3-Aminophenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile (Steps 4 to 5), Compound 2 (Step 1 And 2) and the preparation scheme of compound 68 (step 8), with o-nitroacetophenone and 5-amino-3-(4-phenoxyphenyl)-1H under conditions well known to those skilled in the art Pyrazole-4-carbonitrile is used as a starting material to give the desired product. 1 H NMR (DMSO-d 6 ) δ 7.48-7.44 (m, 2H), 7.40-7.33 (m, 2H), 7.15-7.09 (m, 1H), 7.05-6.97 (m, 4H), 6.92 (td, J = 8.0, 1.2 Hz, 1H), 6.75 (br s, 1H), 6.64 (dd, J = 8.0, 1.2 Hz, 1H), 6.46 (td, J = 7.6, 1.2 Hz, 1H), 6.23 (dd, J = 7.6, 1.2 Hz, 1H), 5.57-5.52 (m, 1H), 5.19 (br s, 2H), 3.27-3.17 (m, 1H), 2.93 (td, J = 2.8, 12.0 Hz, 1H), 2.21-2.11 (m, 1H), 2.10-2.05 (m, 1H). MS (ESI) m/e [M+1] + 426.0.
通過手性製備高效液相色譜法，化合物5被分離成兩個對映立體異構體5a(峰1，R或S，在手性分析的保留時間為7.30min)，和5b(峰2，S或R，在手性分析的保留時間為9.68min)。手性分離條件如下所示：
手性分析條件如下所示：
化合物6：7-(2-丙烯醯氨基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 6 : 7-(2-Propylaminophenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3 -Procarbamide
目標產物的製備以化合物5和丙烯醯氯為起始原料，與化合物3的步驟類似。1H NMR(DMSO-d6)δ 9.81(s,1H),7.50-7.32(m,5H),7.26(td,J=7.6,1.2Hz,1H),7.21-7.08(m,2H),7.04-6.98(m,4H),6.79(s,1H),6.60(dd,J=7.6,1.2Hz,1H),6.50(dd,J=17.0,10.2Hz,1H),6.24(dd,J=17.0,1.9Hz,1H),5.77-5.74(m,2H),3.26-3.22(m,1H),2.98-2.92(m,1H),2.32-2.25(m,1H),1.96-1.93(m,1H).MS(ESI)m/e[M+1]+ 480。 The preparation of the target product was carried out starting from compound 5 and acrylonitrile chloride, similar to the procedure of compound 3 . 1 H NMR (DMSO-d 6 ) δ 9.81 (s, 1H), 7.50-7.32 (m, 5H), 7.26 (td, J = 7.6, 1.2 Hz, 1H), 7.21-7.08 (m, 2H), 7.04 -6.98 (m, 4H), 6.79 (s, 1H), 6.60 (dd, J = 7.6, 1.2 Hz, 1H), 6.50 (dd, J = 17.0, 10.2 Hz, 1H), 6.24 (dd, J =17.0) , 1.9 Hz, 1H), 5.77-5.74 (m, 2H), 3.26-3.22 (m, 1H), 2.98-2.92 (m, 1H), 2.32-2.25 (m, 1H), 1.96-1.93 (m, 1H) ). MS (ESI) m / e [M + 1] + 480.
由5a和5b通過製備化合物3的類似方法，分別得到化合物6a(峰1，R或S，在手性分析的保留時間為4.02min)和6b(峰2，S或R，在手性分析的保留時間為6.68min)。 A similar method for the preparation of compound 3 from 5a and 5b gave compound 6a (peak 1, R or S, retention time in chiral analysis 4.02 min) and 6b (peak 2, S or R, in chiral analysis). The retention time is 6.68min).
手性分析條件如下所示：
實施例2：化合物7和8的合成Example 2: Synthesis of Compounds 7 and 8
化合物7：7-(3-氨基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 7 : 7-(3-Aminophenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide
向7-(3-氨基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈(4mg,0.01mmol)的DCE(2mL)溶液加入活性MnO2(100mg,1.15mmol)。混合物在75℃攪拌2小時，過 濾。濾液用製備矽膠板(DCM/CH3OH=10/1)純化得到黃色固體7-(3-氨基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲醯胺(2mg,50%)。1H NMR(400MHz,CD3OD-d4和CDCl3-d1)δ 8.62(d,J=4.4Hz,1H),7.88(d,J=8.8Hz,2H),7.41-7.39(m,2H),7.31-7.26(m,3H),7.11(d,J=4.4Hz,1H),7.10-7.04(m,1H),7.01-6.97(m,4H),6.90-6.86(m,1H)。 To a solution of 7-(3-aminophenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile (4 mg, 0.01 mmol) in DCM (2 mL) Active MnO 2 (100 mg, 1.15 mmol) was added. The mixture was stirred at 75 ° C for 2 hours and filtered. The filtrate was purified by preparative silica gel (DCM/CH 3 OH = 10/1) to give a yellow solid 7-(3-aminophenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a Pyrimidine-3-carbamide (2 mg, 50%). 1 H NMR (400 MHz, CD 3 OD-d 4 and CDCl 3 -d 1 ) δ 8.62 (d, J = 4.4 Hz, 1H), 7.88 (d, J = 8.8 Hz, 2H), 7.41-7.39 (m, 2H), 7.31-7.26 (m, 3H), 7.11 (d, J = 4.4 Hz, 1H), 7.10-7.04 (m, 1H), 7.01-6.97 (m, 4H), 6.90-6.86 (m, 1H) .
化合物8：7-(3-丙烯醯胺基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 8 : 7-(3-Acrylaminophenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide
向7-(3-氨基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲醯胺(100mg,0.24mmol)的DCM(10mL)溶液中，依次加入TEA(3滴)，丙烯醯氯(32mg,0.36mmol)。混合物室溫攪拌1分鐘，之後加入到DCM(50mL)和飽和食鹽水(50mL)中。從水層中分離出有機層，Na2SO4乾燥，濃縮，製備矽膠板(DCM/CH3OH=10/1)純化得到白色固體7-(3-丙烯醯胺基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲醯胺(12mg,11%)。1H NMR(400MHz,DMSO-d6)δ 10.42(s,1H),8.80(d,J=4.4Hz,1H),8.47(s,1H),8.07(s,1H),7.95(d,J=8.8Hz,2H),7.89(d,J=8.4Hz,1H),7.81(d,J=8.0Hz,1H),7.56(t,J=8.0Hz,1H),7.47(br s,1H),7.42-7.37(m,3H),7.15(t,J= 7.6Hz,1H),7.06(d,J=8.0Hz,2H),7.01(d,J=8.8Hz,2H),6.44(dd,J=10.1,16.9Hz,1H),6.26(dd,J=1.6,16.9Hz,1H),5.76(dd,J=1.6,10.1Hz,1H).MS(ESI)m/e[M+1]+ 476。 To 7-(3-Aminophenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (100 mg, 0.24 mmol) in DCM (10 mL) To the solution, TEA (3 drops), propylene hydrazine chloride (32 mg, 0.36 mmol) were added in that order. The mixture was stirred at room temperature for 1 min then added to DCM (50 mL) and brine. Separated from the aqueous layer, the organic layer was dried Na 2 SO 4, concentrated, prepared silicone plate (DCM / CH 3 OH = 10 /1) to give a white solid 7- (3-acrylamide-yl) -2- (4-Phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (12 mg, 11%). 1 H NMR (400MHz, DMSO- d 6) δ 10.42 (s, 1H), 8.80 (d, J = 4.4Hz, 1H), 8.47 (s, 1H), 8.07 (s, 1H), 7.95 (d, J = 8.8 Hz, 2H), 7.89 (d, J = 8.4 Hz, 1H), 7.81 (d, J = 8.0 Hz, 1H), 7.56 (t, J = 8.0 Hz, 1H), 7.47 (br s, 1H) , 7.42 - 7.37 (m, 3H), 7.15 (t, J = 7.6 Hz, 1H), 7.06 (d, J = 8.0 Hz, 2H), 7.01 (d, J = 8.8 Hz, 2H), 6.44 (dd, J =10.1, 16.9 Hz, 1H), 6.26 (dd, J = 1.6, 16.9 Hz, 1H), 5.76 (dd, J = 1.6, 10.1 Hz, 1H). MS (ESI) m/e [M+1] + 476.
實施例3：化合物9-10的合成Example 3: Synthesis of Compound 9-10
化合物9：7-(2-氨基苯基)-2-(4-(苄氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 9 : 7-(2-Aminophenyl)-2-(4-(benzyloxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3 -Procarbamide
步驟1,2,3：5-氨基-3-(4-(苄氧基)苯基)-1H-吡唑-4-甲腈 Step 1, 2, 3: 5-amino-3-(4-(benzyloxy)phenyl)-1H-pyrazole-4-carbonitrile
根據5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈的製備方案(步驟1到3)，以4-(苄氧基)苯甲酸為原料，在本領域普通技術人員熟知的條件下，得到目標產物。1H NMR(400MHz,DMSO-d6)δ 12.01(s,1H),7.72(d,J=8.8Hz,2H),7.50-7.44(m,2H),7.44-7.37(m,2H),7.36-7.32(m,1H),7.11(d,J=7.2Hz,2H),6.39(br s,2H),5.16(s,2H)。 According to the preparation scheme of 5-amino-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitrile (steps 1 to 3), 4-(benzyloxy)benzoic acid is used as a raw material, The desired product is obtained under conditions well known to those of ordinary skill in the art. 1 H NMR (400MHz, DMSO- d 6) δ 12.01 (s, 1H), 7.72 (d, J = 8.8Hz, 2H), 7.50-7.44 (m, 2H), 7.44-7.37 (m, 2H), 7.36 -7.32 (m, 1H), 7.11 (d, J = 7.2 Hz, 2H), 6.39 (br s, 2H), 5.16 (s, 2H).
步驟4,5：2-(4-(苄氧基)苯基)-7-(2-硝基苯基)吡唑並[1,5-a]嘧啶-3-甲腈 Step 4, 5: 2-(4-(Benzyloxy)phenyl)-7-(2-nitrophenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile
根據N-(3-(3-氰基-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-7-基)苯基)乙醯胺的製備方案(步驟4和5)，以1-(2-硝基苯基)乙酮為起始原料，在本領域普通技術人員熟知的條件下，得到目標產物。 Preparation scheme according to N- (3-(3-cyano-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidin-7-yl)phenyl)acetamide (step 4 and 5), starting from 1-(2-nitrophenyl)ethanone, the desired product is obtained under conditions well known to those skilled in the art.
步驟6：7-(2-氨基苯基)-2-(4-羥基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈 Step 6: 7-(2-Aminophenyl)-2-(4-hydroxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carbonitrile
向2-(4-(苄氧基)苯基)-7-(2-硝基苯基)吡唑並[1,5-a]嘧啶-3-甲腈的CH3OH(20mL)和DCM(20mL)溶液中加10% w/w Pd/C(300mg)。該混合物在氫氣條件下室溫攪拌16小時。過濾，矽膠層析柱(DCM/CH3OH洗脫)純化，得到黃色固體7-(2-氨基苯基)-2-(4-羥基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈(0.92g,62%)。MS(ESI,m/e)[M+1]+ 331.9。 2- (4- (benzyloxy) phenyl) -7- (2-nitrophenyl) pyrazolo [1,5-a] pyrimidine-3-carbonitrile CH 3 OH (20mL) and DCM (20 mL) was added 10% w/w Pd/C (300 mg). The mixture was stirred at room temperature under hydrogen for 16 hours. Filtration and purification on a silica gel column (DCM/CH 3 OH elution) afforded 7-(2-aminophenyl)-2-(4-hydroxyphenyl)-4,5,6,7-tetrahydro as a yellow solid. Pyrazolo[1,5-a]pyrimidine-3-carbonitrile (0.92 g, 62%). MS (ESI, m/e) [M+1] + 331.9.
步驟7：7-(2-氨基苯基)-2-(4-(苄氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈 Step 7: 7-(2-Aminophenyl)-2-(4-(benzyloxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3 -carbonitrile
向7-(2-氨基苯基)-2-(4-羥基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈(331mg,1.0mmol)的丙酮(10mL)溶液中加入苄溴(204mg,1.2mmol)和K2CO3(276mg,2.0mmol)，混合物室溫攪拌16小時。加入丙酮(50mL)，過濾。濾液濃縮，得到黃色固體7-(2-氨基苯基)-2-(4-(苄氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈(400mg,95%)。MS(ESI)m/e[M+1]+ 421.9。 To 7-(2-aminophenyl)-2-(4-hydroxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carbonitrile (331 mg, Benzyl bromide (204 mg, 1.2 mmol) and K 2 CO 3 (276 mg, 2.0 mmol) were added to aq. Acetone (50 mL) was added and filtered. The filtrate was concentrated to give 7-(2-aminophenyl)-2-(4-(benzyloxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a] as a yellow solid. Pyrimidine-3-carbonitrile (400 mg, 95%). MS (ESI) m / e [ M + 1] + 421.9.
步驟8：7-(2-氨基苯基)-2-(4-(苄氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 8: 7-(2-Aminophenyl)-2-(4-(benzyloxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3 -Procarbamide
目標產物由7-(2-氨基苯基)-2-(4-(苄氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈製備，與化合物2的步驟2類似。1H NMR(400MHz,DMSO-d6)δ 7.47-7.33(m,7H),7.06(d,J=8.4Hz,2H),6.96(t,J=7.6Hz,1H),6.75(br s,1H),6.68(d,J=7.6Hz,1H),6.50(t,J=7.6Hz,1H),6.28(d,J=7.6Hz,1H),5.59-5.54(m,1H),5.19(br s,2H),5.12(s, 2H),3.30-3.20(m,1H),3.02-2.92(m,1H),2.25-2.14(m,1H),2.13-2.03(m,1H).MS(ESI)m/e[M+1]+ 439.9。 The target product consists of 7-(2-aminophenyl)-2-(4-(benzyloxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3 - Preparation of carbonitrile, similar to step 2 of compound 2 . 1 H NMR (400MHz, DMSO- d 6) δ 7.47-7.33 (m, 7H), 7.06 (d, J = 8.4Hz, 2H), 6.96 (t, J = 7.6Hz, 1H), 6.75 (br s, 1H), 6.68 (d, J = 7.6 Hz, 1H), 6.50 (t, J = 7.6 Hz, 1H), 6.28 (d, J = 7.6 Hz, 1H), 5.59-5.54 (m, 1H), 5.19 ( Br s,2H), 5.12(s, 2H), 3.30-3.20(m,1H), 3.02-2.92(m,1H), 2.25-2.14(m,1H),2.13-2.03(m,1H).MS (ESI) m/e [M+1] + 439.9.
化合物10：7-(2-丙烯醯胺基苯基)-2-(4-(苄氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 10 : 7-(2-Acrylaminophenyl)-2-(4-(benzyloxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a] Pyrimidine-3-carboxamide
目標產物由7-(2-氨基苯基)-2-(4-(苄氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺和丙烯醯氯製備，與化合物8的步驟類似。1H NMR(400MHz,DMSO-d6)δ 9.84(s,1H),7.46-7.37(m,7H),7.33-7.28(m,2H),7.21(t,J=7.6Hz,1H),7.06(d,J=8.8Hz,2H),6.81(br s,1H),6.64(d,J=7.6Hz,1H),6.53(dd,J=10.3,16.8Hz,1H),6.27(d,J=1.8,16.8Hz,1H),5.59-5.57(m,2H),5.12(s,2H),3.30-3.26(m,1H),3.04-2.92(m,1H),2.35-2.27(m,1H),1.95-1.97(m,1H).MS(ESI)m/e[M+1]+ 493.9。 The target product consists of 7-(2-aminophenyl)-2-(4-(benzyloxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3 - Preparation of formamide and propylene chloride, similar to the procedure of compound 8 . 1 H NMR (400MHz, DMSO- d 6) δ 9.84 (s, 1H), 7.46-7.37 (m, 7H), 7.33-7.28 (m, 2H), 7.21 (t, J = 7.6Hz, 1H), 7.06 (d, J = 8.8 Hz, 2H), 6.81 (br s, 1H), 6.64 (d, J = 7.6 Hz, 1H), 6.53 (dd, J = 10.3, 16.8 Hz, 1H), 6.27 (d, J =1.8,16.8Hz,1H),5.59-5.57(m,2H),5.12(s,2H), 3.30-3.26(m,1H),3.04-2.92(m,1H),2.35-2.27(m,1H ), 1.95-1.97 (m, 1H). MS (ESI) m/e [M+1] + 493.9.
通過手性製備高效液相色譜法，化合物10被分離成兩個對映立體異構體10a(峰1，R或S，在手性分析的保留時間為3.15min)，和10b(峰2，S或R，在手性分析的保留時間為3.91min)。手性分離條件如下所示：
手性分析條件如下所示：
實施例4：化合物11-12的合成Example 4: Synthesis of Compound 11-12
化合物11：7-(2-氨基苯基)-2-(4-(4-氟苯氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 11 : 7-(2-Aminophenyl)-2-(4-(4-fluorophenoxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a] Pyrimidine-3-carboxamide
步驟1：2-(3-氰基-2-(4-羥基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)苯氨基甲酸苄酯 Step 1: 2-(3-Cyano-2-(4-hydroxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-7-yl)phenylcarbamic acid Benzyl ester
向7-(2-氨基苯基)-2-(4-羥基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈(730mg,2.21mmol)的THF(30mL)溶液中，依次加入K2CO3(610mg,4.42mmol)，CbzCl(564mg,3.32mmol)。65℃攪拌16小時，混合物減壓濃縮。殘餘物加入DCM(150mL)和飽和食鹽水(150mL)中。將有機層從水層分離，Na2SO4乾燥，矽膠層析柱純化，用DCM/CH3OH洗脫，得到白色固體2-(3-氰基-2-(4-羥基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)苯氨基甲酸苄酯(370mg,62%)。1H NMR(400MHz,DMSO-d6)δ 9.71(s,1H),9.33(s,1H),7.66(s,1H),7.58(d,J=8.4Hz,2H),7.44-7.28(m,7H),7.17(t,J=7.6Hz,1H),6.79(d,J=8.4Hz,2H),6.59(d,J=7.6Hz,1H),5.82-5.77(m,1H),5.17(s,2H),3.25-3.18(m,1H),2.97-2.87(m,1H),2.36-2.24(m,1H),2.08-2.00(m,1H)。 To 7-(2-aminophenyl)-2-(4-hydroxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carbonitrile (730 mg, 2.21 mmol) in THF (30mL) solution was added sequentially K 2 CO 3 (610mg, 4.42mmol ), CbzCl (564mg, 3.32mmol). After stirring at 65 ° C for 16 hours, the mixture was concentrated under reduced pressure. The residue was taken up in DCM (150 mL) and brine. The organic layer was separated from the aqueous layer, dried 2 SO 4 Na, purified by column chromatography on silica gel, eluting with DCM / CH 3 OH, to give a white solid of 2- (3-cyano-2- (4-hydroxyphenyl) - Benzyl 4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-7-yl)phenylcarbamate (370 mg, 62%). 1 H NMR (400MHz, DMSO- d6) δ 9.71 (s, 1H), 9.33 (s, 1H), 7.66 (s, 1H), 7.58 (d, J = 8.4Hz, 2H), 7.44-7.28 (m, 7H), 7.17 (t, J = 7.6 Hz, 1H), 6.79 (d, J = 8.4 Hz, 2H), 6.59 (d, J = 7.6 Hz, 1H), 5.82-5.77 (m, 1H), 5.17 ( s, 2H), 3.25-3.18 (m, 1H), 2.97-2.87 (m, 1H), 2.36-2.24 (m, 1H), 2.08-2.00 (m, 1H).
步驟2：2-(3-氰基-2-(4-(4-氟苯氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)苯氨基甲酸苄酯 Step 2: 2-(3-Cyano-2-(4-(4-fluorophenoxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine- Benzyl 7-yl) phenyl carbamate
向2-(3-氰基-2-(4-羥基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)苯氨基甲酸苄酯(370mg,0.8mmol)的DCM(20mL)溶液中加入4-氟苯硼酸(167mg,1.2mmol)，TEA(162mg,1.6mmol)和Cu(OAc)2(216mg,1.2mmol)。室溫攪拌16小時，向混合物中加入DCM(100mL),CH3OH(10mL)和飽和食鹽水(100mL)。將有機層從水層分離，用飽和食鹽水(100mL×2)洗滌，Na2SO4乾燥，矽膠層析柱純化，DCM/CH3OH洗脫得到白色固體2-(3-氰基-2-(4-(4-氟苯氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)苯氨基甲酸苄酯(334mg,75%)。1H NMR(400MHz,DMSO-d6)δ 9.34(s,1H),7.76-7.74(m,3H),7.45-7.10(m,12H),7.02(d,J=8.4Hz,2H),6.59(d,J=8.0Hz,1H),5.85-5.80(m,1H),5.17(s,2H),3.25-3.18(m,1H),2.97-2.87(m,1H),2.36-2.24(m,1H),2.08-2.00(m,1H)。 Benzyl 2-(3-cyano-2-(4-hydroxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-7-yl)benzate (370mg, 0.8mmol) in DCM (20mL) was added a solution of 4-fluorophenyl boronic acid (167mg, 1.2mmol), TEA ( 162mg, 1.6mmol) and Cu (OAc) 2 (216mg, 1.2mmol). Was stirred at room temperature for 16 hours, DCM (100mL) added to the mixture, CH 3 OH (10mL) and saturated brine (100mL). The organic layer was separated from the aqueous layer, washed with saturated brine (100mL × 2), dried Na 2 SO 4, purified by column chromatography on silica gel, DCM / CH 3 OH to afford a white solid 2- (3-cyano-2 -(4-(4-Fluorophenoxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-7-yl)benzidinecarboxylate (334 mg, 75%). 1 H NMR (400MHz, DMSO- d6) δ 9.34 (s, 1H), 7.76-7.74 (m, 3H), 7.45-7.10 (m, 12H), 7.02 (d, J = 8.4Hz, 2H), 6.59 ( d, J = 8.0 Hz, 1H), 5.85-5.80 (m, 1H), 5.17 (s, 2H), 3.25-3.18 (m, 1H), 2.97-2.87 (m, 1H), 2.36-2.24 (m, 1H), 2.08-2.00 (m, 1H).
步驟3：2-(3-氨基甲醯基-2-(4-(4-氟苯基氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)苯氨基甲酸苄酯 Step 3: 2-(3-Carbamino-2-(4-(4-fluorophenyloxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a Benzyl pyridin-7-yl)phenylcarbamate
目標產物由2-(3-氰基-2-(4-(4-氟苯氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)苯氨基甲酸苄酯製備，與化合物2的步驟2類似。MS(ESI)m/e[M+1]+ 577.9。 The target product consists of 2-(3-cyano-2-(4-(4-fluorophenoxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine- The preparation of benzyl 7-yl)benzate is similar to Step 2 of Compound 2 . MS (ESI) m / e [M+1] + 577.9.
步驟4：7-(2-氨基苯基)-2-(4-(4-氟苯基氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 4: 7-(2-Aminophenyl)-2-(4-(4-fluorophenyloxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a Pyrimidine-3-carboxamide
向2-(3-氨基甲醯基-2-(4-(4-氟苯基氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)苯氨基甲酸苄酯(100mg,0.17mmol)的DCM(5mL)和MeOH(5mL)溶液中加入10% w/w Pd/C(50mg)。在H2條件下，室溫攪拌16小時，混合物過濾，濾餅用DCM/CH3OH(1/1,50mL)洗滌。濾液濃縮，矽膠層析柱純化，DCM/CH3OH洗脫，得到白色固體7-(2-氨基苯基)-2-(4-(4-氟苯基氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺(10mg,13%)。1H NMR(400MHz,CD3OD-d4)δ 7.39(d,2H,J=8.8Hz),7.02-6.91(m,7H),6.66(d,J=7.6Hz,1H),6.53(t,J=7.6Hz,1H),6.33(d,J=7.6Hz,1H),5.54-5.50(m, 1H),3.30-3.24(m,1H),3.12-3.06(m,1H),2.31-2.20(m,2H).MS(ESI)m/e[M+1]+ 443.9。 To 2-(3-carbamoyl-2-(4-(4-fluorophenyloxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine To a solution of benzyl phenyl-phenylcarbamate (100 mg, 0.17 mmol) in DCM (5 mL) and MeOH (5 mL), 10% w/w Pd/C (50 mg). Under conditions of H 2, stirred at room temperature for 16 hours the mixture was filtered and the cake was washed with DCM / CH 3 OH (1 / 1,50mL). The filtrate was concentrated, purified on a silica gel column eluting with DCM/CH 3 EtOAc to afford 7-(2-aminophenyl)-2-(4-(4-fluorophenyloxy)phenyl)-4 as a white solid. 5,6,7-Tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide (10 mg, 13%). 1 H NMR (400 MHz, CD 3 OD-d 4 ) δ 7.39 (d, 2H, J = 8.8 Hz), 7.02-6.91 (m, 7H), 6.66 (d, J = 7.6 Hz, 1H), 6. , J = 7.6 Hz, 1H), 6.33 (d, J = 7.6 Hz, 1H), 5.54-5.50 (m, 1H), 3.30-3.24 (m, 1H), 3.12-3.06 (m, 1H), 2.31 2.20 (m, 2H). MS (ESI) m/e [M+1] + 443.9.
化合物12：7-(2-丙烯醯胺基苯基)-2-(4-(4-氟苯基氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 12 : 7-(2-Acrylaminophenyl)-2-(4-(4-fluorophenyloxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1, 5-a]pyrimidine-3-carboxamide
目標產物由7-(2-氨基苯基)-2-(4-(4-氟苯基氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺和丙烯醯氯製備，與化合物8的步驟類似。1H NMR(400MHz,CD3OD-d4)δ 7.47(d,J=8.4Hz,2H),7.39(t,J=7.6Hz,1H),7.33(t,J=7.6Hz,1H),7.26(t,J=7.6Hz,1H),7.12-7.00(m,6H),6.81(d,J=8.0Hz,1H),6.53-6.46(m,1H),6.39-6.35(m,1H),5.87-5.76(m,1H),5.73-5.69(m,1 H),3.36-3.30(m,1H),3.22-3.17(m,1H),2.45-2.39(m,1 H),2.17-2.14(m,1H).MS(ESI)m/e[M+1]+ 497.9。 The target product consists of 7-(2-aminophenyl)-2-(4-(4-fluorophenyloxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a The preparation of pyrimidine-3-carbamide and acrylonitrile chloride is similar to the procedure of compound 8 . 1 H NMR (400 MHz, CD 3 OD-d 4 ) δ 7.47 (d, J = 8.4 Hz, 2H), 7.39 (t, J = 7.6 Hz, 1H), 7.33 (t, J = 7.6 Hz, 1H), 7.26(t, J = 7.6 Hz, 1H), 7.12-7.00 (m, 6H), 6.81 (d, J = 8.0 Hz, 1H), 6.53-6.46 (m, 1H), 6.39-6.35 (m, 1H) , 5.87-5.76 (m, 1H), 5.73-5.69 (m, 1 H), 3.36-3.30 (m, 1H), 3.22-3.17 (m, 1H), 2.45-2.39 (m, 1 H), 2.17- 2.14 (m, 1H). MS (ESI) m / e [M + 1] + 497.9.
實施例5：化合物13-14的合成Example 5: Synthesis of Compound 13-14
化合物13：7-(2-(甲氨基)苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 13 : 7-(2-(methylamino)phenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine- 3-methylamine
步驟1：N-(2-(3-氰基-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-7-基)苯基)-2,2,2-三氟乙醯胺 Step 1: N- (2-(3-Cyano-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidin-7-yl)phenyl)-2,2,2 -trifluoroacetamide
向7-(2-氨基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈(60mg,0.15mmol)的DCM(5mL)溶液中加入DIEA(3滴)和三氟醋酸酐(3滴)。反應混合物室溫攪拌2小時，然後加入到水(20mL)和DCM(20mL)中。有機層旋幹得到黃色固體粗品(50mg,67%)，未經進一步純化直接用於下步反應。 To a solution of 7-(2-aminophenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile (60 mg, 0.15 mmol) in DCM (5 mL) DIEA (3 drops) and trifluoroacetic anhydride (3 drops) were added. The reaction mixture was stirred at room temperature for 2 hr then added water (20 mL) and DCM (20 mL). The organic layer was dried <RTI ID=0.0>
步驟2：7-(2-(甲氨基)苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈 Step 2: 7-(2-(Methylamino)phenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile
向N-(2-(3-氰基-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-7-基)苯基)-2,2,2-三氟乙醯胺(50mg,0.1mmol)的丙酮(5mL)溶液中，加入KOH(11.2mg,0.2mmol)和CH3I(0.5mL)。反應混合物室溫攪拌15小時，濃縮除去丙酮。殘餘物加入到水(20 mL)和EA(20mL)中。有機層旋幹，製備矽膠板(PE/EA=2/1)純化得到白色固體產品(15mg,37%)。1H NMR(DMSO-d6)δ 8.82(d,J=4.4Hz,1H),7.97(d,J=8.8Hz,2H),7.47-7.38(m,4H),7.31(dd,J=1.6,7.2Hz,1H),7.23-7.17(m,3H),7.11(d,J=8.8Hz,2H),6.77-6.69(m,2H),5.35-5.31(m,1H),2.68(d,J=4.8Hz,3H).MS(ESI)m/e[M+1]+ 417.9。 To N- (2-(3-cyano-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidin-7-yl)phenyl)-2,2,2-tri fluorine as acetamide (50mg, 0.1mmol) in acetone (5mL) was added KOH (11.2mg, 0.2mmol) and CH 3 I (0.5mL). The reaction mixture was stirred at room temperature for 15 hours and concentrated to remove acetone. The residue was taken into water (20 mL) and EA (20 mL). The organic layer was dried with EtOAc (EtOAc/EtOAc (EtOAc) 1 H NMR (DMSO-d6) δ 8.82 (d, J = 4.4 Hz, 1H), 7.97 (d, J = 8.8 Hz, 2H), 7.47-7.38 (m, 4H), 7.31 (dd, J = 1.6, 7.2 Hz, 1H), 7.23-7.17 (m, 3H), 7.11 (d, J = 8.8 Hz, 2H), 6.77-6.69 (m, 2H), 5.35-5.31 (m, 1H), 2.68 (d, J = 4.8 Hz, 3H). MS (ESI) m/e [M+1] + 417.9.
步驟3：7-(2-(甲氨基)苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈 Step 3: 7-(2-(Methylamino)phenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine- 3-carbonitrile
向7-(2-(甲氨基)苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈(250mg,0.6mmol)的EtOH(10mL)溶液中加入硼氫化鈉(100mg)。反應混合物室溫攪拌15小時，濃縮除去EtOH。殘餘物加入到水(30mL)和EA(30mL)中，有機層濃縮。殘餘物溶解在MeOH(10mL)中，再加入10% w/w Pd/C(50mg)。反應混合物室溫1個大氣壓的氫氣下攪拌15小時。然後，過濾混合物，旋幹濾液，矽膠層析柱(PE/EA=1/1)純化，得到白色固體產品(144mg,yield：56.5%)。MS(ESI)m/e[M+1]+ 421.9。 To 7-(2-(methylamino)phenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile (250 mg, 0.6 mmol) in EtOH ( Sodium borohydride (100 mg) was added to the solution of 10 mL). The reaction mixture was stirred at room temperature for 15 hours and concentrated to remove EtOH. The residue was taken into water (30 mL) EtOAc The residue was dissolved in MeOH (10 mL) and then 10% w/w Pd / C (50 mg). The reaction mixture was stirred at room temperature under 1 atmosphere of hydrogen for 15 hours. Then, the mixture was filtered, and the filtrate was evaporated to dryness eluted eluted eluted eluted elution MS (ESI) m / e [ M + 1] + 421.9.
步驟4：7-(2-(甲氨基)苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 4: 7-(2-(Methylamino)phenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine- 3-methylamine
目標產物由7-(2-(甲氨基)苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈製備，與化合物2的步驟2類似。1H NMR(DMSO-d6)δ 7.49(d,J=8.8Hz,2H),7.44-7.35(m,2H),7.18-7.01(m,6H),6.76(s,1H),6.58(d,J=7.6Hz,2H),6.54(t,J=7.6Hz,1H),6.28(dd,J=1.2,7.6Hz,1H),5.60-5.57(m,1H),5.50-5.49(m,1H),3.29-3.24(m,1H),2.98-2.93(m,1H),2.76(d,J=4.8Hz,3H),2.24-2.18(m,1H),2.06-2.03(m,1H).MS(ESI)m/e[M+1]+ 439.8。 The target product consists of 7-(2-(methylamino)phenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine- 3-carbonitrile preparation, similar to step 2 of compound 2 . 1 H NMR (DMSO-d6) δ 7.49 (d, J = 8.8 Hz, 2H), 7.44 - 7.35 (m, 2H), 7.18 - 7.01 (m, 6H), 6.76 (s, 1H), 6.58 (d, J = 7.6 Hz, 2H), 6.54 (t, J = 7.6 Hz, 1H), 6.28 (dd, J = 1.2, 7.6 Hz, 1H), 5.60-5.57 (m, 1H), 5.50-5.49 (m, 1H) ), 3.29-3.24 (m, 1H), 2.98-2.93 (m, 1H), 2.76 (d, J = 4.8 Hz, 3H), 2.24-2.18 (m, 1H), 2.06-2.03 (m, 1H). MS (ESI) m / e [M+1] + 439.8.
化合物14：7-(2-(N-甲基丙烯醯胺基)苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 14 : 7-(2-( N -methylpropenylamino)phenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1, 5-a]pyrimidine-3-carboxamide
目標產物由7-(2-(甲氨基)苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺和丙烯醯氯製備，與化合物8的步驟類似。1H NMR(DMSO-d6)δ 7.45-7.37(m,6H),7.29-7.24(m,1H),7.15(t,J=7.6Hz,1H),7.06-7.00(m,5H),6.81(d,J=7.6Hz,1H),6.25-6.16(m,2H),5.94-5.87(m,1H), 5.63-5.51(m,1H),5.41-5.35(m,1H),3.41-3.22(m,5H),2.41-1.97(m,2H).MS(ESI)m/e[M+1]+ 493.9。 The target product consists of 7-(2-(methylamino)phenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine- 3-Methylamine and acrylonitrile chloride were prepared in a similar manner to Compound 8 . 1 H NMR (DMSO-d6) δ 7.45-7.37 (m, 6H), 7.29-7.24 (m, 1H), 7.15 (t, J = 7.6 Hz, 1H), 7.06-7.00 (m, 5H), 6.81 ( d, J = 7.6 Hz, 1H), 6.25-6.16 (m, 2H), 5.94 - 5.87 (m, 1H), 5.63-5.51 (m, 1H), 5.41-5.35 (m, 1H), 3.41-3.22 ( m, 5H), 2.41-1.97 (m, 2H). MS (ESI) m/e [M+1] + 493.9.
實施例6：化合物15-16的合成Example 6: Synthesis of Compound 15-16
化合物15：2-(4-(4-氟基苯氧基)苯基)-7-(2-(甲基氨基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 15 : 2-(4-(4-Fluorophenoxy)phenyl)-7-(2-(methylamino)phenyl)-4,5,6,7-tetrahydropyrazolo[1 ,5-a]pyrimidine-3-carboxamide
步驟1：N-(2-(3-氰基-2-(4-羥基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)苯基)-2,2,2-三氟乙醯胺 Step 1: N- (2-(3-Cyano-2-(4-hydroxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-7-yl) Phenyl)-2,2,2-trifluoroacetamide
向7-(2-氨基苯基)-2-(4-羥基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈(33.1mg,0.1mmol)的二氯甲烷(10mL)溶液中加入三氟醋酸酐(1滴)和DIEA(1滴)。室溫攪拌1小時後，將混合物加入到二氯甲烷(10mL)和飽和食鹽水(10mL)中。分離出有機層，Na2SO4乾燥，濃縮，得到黃色固體N-(2-(3-氰基-2-(4-羥基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)苯基)-2,2,2-三氟乙醯胺(40mg,93%)。MS(ESI)m/e[M+1]+ 427.8。 To 7-(2-aminophenyl)-2-(4-hydroxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carbonitrile (33.1 mg Trifluoroacetic anhydride (1 drop) and DIEA (1 drop) were added to a solution of 0.1 mmol) in dichloromethane (10 mL). After stirring at room temperature for 1 hour, the mixture was added to dichloromethane (10 mL) and brine (10 mL). The organic layer was separated, dried Na 2 SO 4, and concentrated to give a yellow solid N - (2- (3- cyano-2- (4-hydroxyphenyl) -4,5,6,7-tetrahydro-pyrazolo [1,5-a]pyrimidin-7-yl)phenyl)-2,2,2-trifluoroacetamide (40 mg, 93%). MS (ESI) m / e [ M + 1] + 427.8.
步驟2：N-(2-(3-氰基-2-(4-(4-氟苯氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)苯基)-2,2,2-三氟乙醯胺 Step 2: N- (2-(3-Cyano-2-(4-(4-fluorophenoxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a Pyrimidine-7-yl)phenyl)-2,2,2-trifluoroacetamide
向N-(2-(3-氰基-2-(4-羥基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)苯基)-2,2,2-三氟乙醯胺(42.7mg,0.10mmol)的二氯甲烷(3mL)溶液中加入4-氟苯硼酸(17mg,0.12mmol)，TEA(21mg,0.2mmol)和Cu(OAc)2(22mg,0.12mmol)。室溫攪拌16小時後，製備矽膠板(DCM/CH3OH=20/1)純化，得到無色油狀物N-(2-(3-氟基-2-(4-(4-氟苯氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)苯基)-2,2,2-三氟乙醯胺(30mg,57%)。1H NMR(400MHz,DMSO-d6)δ 11.33(s,1H),7.90-7.77(m,3H),7.52-7.37(m,3H),7.34-7.27(m,2H),7.22-7.14(m,2H),7.10(d,J=8.8Hz,2H),6.80(d,J=6.4Hz,1H),5.80-5.75(m,1H),3.37-3.28(m,1H),3.09-2.95(m,1H),2.50-2.37(m,1H),2.10-1.95(m,1H).MS(ESI)m/e[M+1]+ 521.8。 To N- (2-(3-cyano-2-(4-hydroxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-7-yl)phenyl To a solution of -2,2,2-trifluoroacetamide (42.7 mg, 0.10 mmol) in dichloromethane (3 mL), EtOAc (EtOAc (EtOAc) Cu(OAc) 2 (22 mg, 0.12 mmol). After stirring at room temperature for 16 hours, the prepared silica gel plate (DCM/CH 3 OH=20/1) was purified to give N- (2-(3-fluoro)-2-(4-(4-fluorophenoxy) as a colorless oil. Phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-7-yl)phenyl)-2,2,2-trifluoroacetamide (30 mg, 57%). 1 H NMR (400MHz, DMSO- d 6) δ 11.33 (s, 1H), 7.90-7.77 (m, 3H), 7.52-7.37 (m, 3H), 7.34-7.27 (m, 2H), 7.22-7.14 ( m, 2H), 7.10 (d, J = 8.8 Hz, 2H), 6.80 (d, J = 6.4 Hz, 1H), 5.80-5.75 (m, 1H), 3.37-3.28 (m, 1H), 3.09-2.95 (m, 1H), 2.50-2.37 (m, 1H), 2.10-1.95 (m, 1H). MS (ESI) m/e [M+1] + 521.8.
步驟3：N-(2-(3-氰基-2-(4-(4-氟苯氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)苯基)-2,2,2-三氟-N-甲基乙醯胺 Step 3: N- (2-(3-Cyano-2-(4-(4-fluorophenoxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a Pyrimidin-7-yl)phenyl)-2,2,2-trifluoro- N -methylacetamide
向N-(2-(3-氰基-2-(4-(4-氟苯氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)苯基)-2,2,2-三氟乙醯胺(187mg,0.36mmol)的丙酮(10mL)溶液中加入K2CO3(100mg,0.72mmol)和CH3I(15滴)。室溫攪拌2小時後，混合物過濾，濃縮濾液，得到黃色固體N-(2-(3-氰基-2-(4-(4-氟苯氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)苯基)-2,2,2-三氟-N-甲基乙醯胺(192mg,100%)。MS(ESI)m/e[M+1]+ 535.8。 To N- (2-(3-cyano-2-(4-(4-fluorophenoxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine 7-yl) phenyl) -2,2,2-trifluoro-acetyl amine (187mg, 0.36mmol) in acetone (10 mL) was added K 2 CO 3 (100mg, 0.72mmol ) and CH 3 I (15 drop). After stirring for 2 hours at room temperature, the mixture was filtered, the filtrate was concentrated to give a yellow solid N - (2- (3- cyano-2- (4- (4-fluorophenoxy) phenyl) -4,5,6, 7-Tetrahydropyrazolo[1,5-a]pyrimidin-7-yl)phenyl)-2,2,2-trifluoro- N -methylacetamide (192 mg, 100%). MS (ESI) m / e [M+1] + 535.8.
步驟4：2-(4-(4-氟苯氧基)苯基)-7-(2-(甲基氨基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 4: 2-(4-(4-Fluorophenoxy)phenyl)-7-(2-(methylamino)phenyl)-4,5,6,7-tetrahydropyrazolo[1, 5-a]pyrimidine-3-carboxamide
目標產物由N-(2-(3-氰基-2-(4-(4-氟苯氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)苯基)-2,2,2-三氟-N-甲基乙醯胺製備，與化合物2的步驟2類似。1H NMR(400MHz,CD3OD-d4)δ 7.39(d,J=8.8Hz,2H),7.09-6.90(m,7H),6.58(d,J=8.0Hz,1H),6.51(t,J=7.2Hz,1H),6.30(d,J=7.2Hz,1H),5.52-5.48(m,1H),3.27-3.24(m,1H),3.11-3.02(m, 1H),2.76(s,3H),2.32-2.10(m,2H).MS(ESI)m/e[M+1]+ 457.9。 The target product consists of N- (2-(3-cyano-2-(4-(4-fluorophenoxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a Preparation of pyrimidin-7-yl)phenyl)-2,2,2-trifluoro- N -methylacetamide, similar to Step 2 of Compound 2 . 1 H NMR (400MHz, CD 3 OD-d 4) δ 7.39 (d, J = 8.8Hz, 2H), 7.09-6.90 (m, 7H), 6.58 (d, J = 8.0Hz, 1H), 6.51 (t , J = 7.2 Hz, 1H), 6.30 (d, J = 7.2 Hz, 1H), 5.52-5.48 (m, 1H), 3.27-3.24 (m, 1H), 3.11-3.02 (m, 1H), 2.76 ( s, 3H), 2.32-2.10 (m, 2H). MS (ESI) m/e [M+1] + 457.9.
化合物16：2-(4-(4-氟苯氧基)苯基)-7-(2-(N-甲基丙烯醯氨基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 16 : 2-(4-(4-fluorophenoxy)phenyl)-7-(2-( N -methylpropenylamino)phenyl)-4,5,6,7-tetrahydropyrazole And [1,5-a]pyrimidine-3-carboxamide
目標產物由2-(4-(4-氟苯氧基)苯基)-7-(2-(甲基氨基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺和丙烯醯氯製備，與化合物8的步驟類似。1H NMR(400MHz,DMSO-d6)δ 7.47-7.37(m,4H),7.30-7.15(m,3H),7.12-7.08(m,2H),6.99(d,J=8.4Hz,2H),6.81(d,J=7.2Hz,1H),6.25-5.85(m,2H),5.63-5.49(m,1H),5.42-5.32(m,1H),3.40-3.20(m,6H),2.45-2.20(m,1H),2.15-1.90(m,1H).MS(ESI)m/e[M+1]+ 511.9。 The target product consists of 2-(4-(4-fluorophenoxy)phenyl)-7-(2-(methylamino)phenyl)-4,5,6,7-tetrahydropyrazolo[1, Preparation of 5-a]pyrimidine-3-carboxamide and acrylonitrile chloride, similar to the procedure of compound 8 . 1 H NMR (400MHz, DMSO- d 6) δ 7.47-7.37 (m, 4H), 7.30-7.15 (m, 3H), 7.12-7.08 (m, 2H), 6.99 (d, J = 8.4Hz, 2H) , 6.81 (d, J = 7.2 Hz, 1H), 6.25-5.85 (m, 2H), 5.63-5.49 (m, 1H), 5.42-5.32 (m, 1H), 3.40-3.20 (m, 6H), 2.45 -2.20 (m, 1H), 2.15 - 1.90 (m, 1H). MS (ESI) m/e [M+1] + 511.9.
實施例7：化合物17-18的合成Example 7: Synthesis of Compound 17-18
化合物17：7-(2-氨基苯基)-2-(4-(環戊氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 17 : 7-(2-Aminophenyl)-2-(4-(cyclopentyloxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine- 3-methylamine
根據化合物9(步驟7和8)的類似製備方案，由7-(2-氨基苯基)-2-(4-羥基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈和溴代環戊烷，在本領域普通技術人員熟知的條件下，得到目標產物。1H NMR(400MHz,DMSO-d6)δ 7.37(d,J=8.8Hz,2H),6.97-6.92(m,3H),6.74(br s,1H),6.67(d,J=7.6Hz,1H),6.49(t,J=7.6Hz,1H),6.27(d,J=7.6Hz,1H),5.58-5.53(m,1H),5.15(s,2H),4.86-4.80(m,1H),3.24-3.27(m,1H),3.02-2.93(m,1H),2.22-2.16(m,1H),2.14-2.08(m,1H),1.98-1.85(m,2H),1.91-1.70(m,4H),1.64-1.54(m,2H).MS(ESI)m/e[M+1]+ 418.0。 According to a similar preparation scheme for compound 9 (steps 7 and 8), from 7-(2-aminophenyl)-2-(4-hydroxyphenyl)-4,5,6,7-tetrahydropyrazolo[1 , 5-a]pyrimidine-3-carbonitrile and bromocyclopentane, the target product is obtained under conditions well known to those skilled in the art. 1 H NMR (400MHz, DMSO- d6) δ 7.37 (d, J = 8.8Hz, 2H), 6.97-6.92 (m, 3H), 6.74 (br s, 1H), 6.67 (d, J = 7.6Hz, 1H ), 6.49 (t, J = 7.6 Hz, 1H), 6.27 (d, J = 7.6 Hz, 1H), 5.58-5.53 (m, 1H), 5.15 (s, 2H), 4.86-4.80 (m, 1H) , 3.24 - 3.27 (m, 1H), 3.02 - 2.93 (m, 1H), 2.22 - 2.16 (m, 1H), 2.14 - 2.08 (m, 1H), 1.98 - 1.85 (m, 2H), 1.91-1.70 ( m, 4H), 1.64-1.54 (m, 2H). MS (ESI) m/e [M+1] + 418.0.
化合物18：7-(2-丙烯醯胺基苯基)-2-(4-(環戊氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 18 : 7-(2-Acrylaminophenyl)-2-(4-(cyclopentyloxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a Pyrimidine-3-carboxamide
目標產物由化合物17和丙烯醯氯製備，與化合物8的步驟類似。1H NMR(400MHz,DMSO-d6)δ 9.83(s,1H),7.45(d,J=7.6Hz,1H),7.37(d,J=8.8Hz,2H)7.29(t,J=7.6Hz,1H),7.21(t,J=7.6Hz,1H),6.93(d,J=8.8Hz,2H),6.81(s,1H),6.64(d,J=7.6Hz,1H),6.53(dd,J=10.2,17.0Hz,1H),6.27(d,J=17.0Hz,1H),5.80-5.77(m,2H),4.86-4.79(m,1H),3.27-3.23(m,1H),3,03-2.94(m,1H),2.36-2.25 (m,1H),1.99-1.91(m,3H),1.75-1.53(m,6H).MS(ESI)m/e[M+1]+ 471.9。 The target product was prepared from compound 17 and acrylonitrile chloride, similar to the procedure of compound 8 . 1 H NMR (400MHz, DMSO- d 6) δ 9.83 (s, 1H), 7.45 (d, J = 7.6Hz, 1H), 7.37 (d, J = 8.8Hz, 2H) 7.29 (t, J = 7.6Hz , 1H), 7.21 (t, J = 7.6 Hz, 1H), 6.93 (d, J = 8.8 Hz, 2H), 6.81 (s, 1H), 6.64 (d, J = 7.6 Hz, 1H), 6.53 (dd , J =10.2, 17.0 Hz, 1H), 6.27 (d, J = 17.0 Hz, 1H), 5.80-5.77 (m, 2H), 4.86-4.79 (m, 1H), 3.27-3.23 (m, 1H), 3,03-2.94 (m,1H), 2.36-2.25 (m,1H), 1.99-1.91 (m,3H),1.75-1.53 (m,6H).MS (ESI) m/e[M+1] + 471.9.
實施例8：化合物19-20的合成Example 8: Synthesis of Compound 19-20
化合物19：7-(2-氨基苯基)-2-(4-(環丙基甲氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 19 : 7-(2-Aminophenyl)-2-(4-(cyclopropylmethoxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a] Pyrimidine-3-carboxamide
步驟1：7-(2-氨基苯基)-2-(4-(環丙基甲氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈 Step 1: 7-(2-Aminophenyl)-2-(4-(cyclopropylmethoxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a] Pyrimidine-3-carbonitrile
向7-(2-氨基苯基)-2-(4-羥基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈(331mg,1.0mmol)在丙酮(15mL)的溶液中，加入(溴甲基)環丙烷(135mg,1.0mmol)和K2CO3(276mg,2.0mmol)。56℃攪拌16hr後，將混合物過濾。濾餅用丙酮(20mL×2)洗滌。將濾液濃縮，得到黃色固體產品(300mg,78%)。MS(ESI)m/e[M+1]+ 386.0。 To 7-(2-aminophenyl)-2-(4-hydroxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carbonitrile (331 mg, 1.0 mmol) In a solution of acetone (15 mL), (bromomethyl)cyclopropane (135 mg, 1.0 mmol) and K 2 CO 3 (276 mg, 2.0 mmol). After stirring at 56 ° C for 16 hr, the mixture was filtered. The filter cake was washed with acetone (20 mL x 2). The filtrate was concentrated to give a yellow solid (300 mg, 78%). MS (ESI) m/e [M+1] + 386.0.
步驟2：7-(2-氨基苯基)-2-(4-(環丙基甲氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 2: 7-(2-Aminophenyl)-2-(4-(cyclopropylmethoxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a] Pyrimidine-3-carboxamide
向7-(2-氨基苯基)-2-(4-(環丙基甲氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈(350mg,0.91mmol)在EtOH(4mL)和DMSO(4mL)的溶液中，加入NaOH水溶液(5N,2mL)和H2O2(2mL)。在60℃下攪拌3hr後，將混合物在水(100mL)和EA(100mL)中分配。將有機層從水層分離，用飽和鹽水(100mL×2)洗滌，硫酸鈉乾燥並濃縮。殘餘物通過矽膠色譜柱(用DCM/MeOH洗脫)，得到白色固體產品(150mg,41%)。1H NMR(400MHz,DMSO-d6)δ 7.37(d,J=8.8Hz,2H),6.97-6.92(m,3H),6.75(br s,1H),6.67(d,J=8.0Hz,1H),6.49(t,J=7.6Hz,1H),6,27(d,J=7.6Hz,1H),5.58-5.54(m,1H),5.16(s,2H),3.83(d,J=7.2Hz,2H),3.27-3.24(m,1H),3.01-2.92(m,1H),2.23-2.13(m,1H),2.13-2.07(m,1H),1.24-1.18(m,1H),0.59-0.54(m,2H),0.34-0.30(m,2H).MS(ESI)m/e[M+1]+ 404.0。 To 7-(2-aminophenyl)-2-(4-(cyclopropylmethoxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine- 3-carbonitrile (350mg, 0.91mmol) in EtOH (4mL) and DMSO (4mL) was added aqueous NaOH (5N, 2mL) and H 2 O 2 (2mL). After stirring at 60 ° C for 3 hr, the mixture was partitioned between water (100 mL) and EA (100 mL). The organic layer was separated from aqueous layer, washed with brine sat. The residue was purified by EtOAc EtOAc EtOAc EtOAc 1 H NMR (400MHz, DMSO- d6) δ 7.37 (d, J = 8.8Hz, 2H), 6.97-6.92 (m, 3H), 6.75 (br s, 1H), 6.67 (d, J = 8.0Hz, 1H ), 6.49 (t, J = 7.6 Hz, 1H), 6, 27 (d, J = 7.6 Hz, 1H), 5.58-5.54 (m, 1H), 5.16 (s, 2H), 3.83 (d, J = 7.2 Hz, 2H), 3.27-3.24 (m, 1H), 3.01-2.92 (m, 1H), 2.23 - 2.13 (m, 1H), 2.13 - 2.07 (m, 1H), 1.24-1.18 (m, 1H) , 0.59-0.54 (m, 2H), 0.34-0.30 (m, 2H) .MS (ESI) m / e [m + 1] + 404.0.
化合物20：7-(2-丙烯醯胺基苯基)-2-(4-(環丙基甲氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 20 : 7-(2-Acrylaminophenyl)-2-(4-(cyclopropylmethoxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5 -a]pyrimidine-3-carboxamide
目標產物由化合物19和丙烯醯氯，製備與化合物8的步驟類似。1H NMR(400MHz,DMSO-d6)δ 9.83(s,1H),7.45(d,J=8.0Hz,1H),7.37(d,J=8.4Hz,2H),7.30(t,J=7.6Hz,1H),7.21(t,J=7.6Hz,1H),6.96(d,J=8.4Hz,2H),6.81(s,1H),6.64(d,J=8.0Hz,1H),6.53(dd,J=10.2,16.7Hz,1H),6.27(d,J=16.7Hz,1H),5.80-5.77(m,2H),3.83(d,J=7.2Hz,2H),3.27-3.23(m,1H),3.03-2.93(m,1H),2.36-2.25(m,1H),2.02-1.91(m,1H),1.23-1.14(m,1H),0.59-0.53(m,2H),0.34-0.29(m,2H).MS(ESI)m/e[M+1]+ 457.9。 The target product was prepared from compound 19 and acrylofluorene chloride in the same manner as compound 8 . 1 H NMR (400MHz, DMSO- d 6) δ 9.83 (s, 1H), 7.45 (d, J = 8.0Hz, 1H), 7.37 (d, J = 8.4Hz, 2H), 7.30 (t, J = 7.6 Hz, 1H), 7.21 (t, J = 7.6 Hz, 1H), 6.96 (d, J = 8.4 Hz, 2H), 6.81 (s, 1H), 6.64 (d, J = 8.0 Hz, 1H), 6.53 ( Dd, J =10.2, 16.7 Hz, 1H), 6.27 (d, J = 16.7 Hz, 1H), 5.80-5.77 (m, 2H), 3.83 (d, J = 7.2 Hz, 2H), 3.27-3.23 (m , 1H), 3.03-2.93 (m, 1H), 2.36-2.25 (m, 1H), 2.02-1.91 (m, 1H), 1.23-1.14 (m, 1H), 0.59-0.53 (m, 2H), 0.34 -0.29 (m, 2H). MS (ESI) m/e [M+1] + 457.9.
通過手性製備高效液相色譜法，化合物20被分離成兩個對映立體異構體20a(峰1，R或S，在手性分析的保留時間為3.03min)，和20b(峰2，S或R，在手性分析的保留時間為3.82min)。手性分離條件如下所示：
手性分析條件如下所示：
實施例9：化合物21-22的合成Example 9: Synthesis of Compound 21-22
化合物21：7-(2-氨基苯基)-2-(4-(2-甲氧基乙氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 21 : 7-(2-Aminophenyl)-2-(4-(2-methoxyethoxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrimidine-3-carboxamide
根據化合物9(步驟7和8)的類似製備方案，由7-(2-氨基苯基)-2-(4-羥基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈和2-溴乙基甲基醚，在本領域普通技術人員熟知的條件下，得到目標產物。1H NMR(400MHz,DMSO-d6)δ 7.38(d,J=8.6 Hz,2H),7.00-6.94(m,3H),6.75(br s,1H),6.69(d,J=7.6Hz,1H),6.51(t,J=7.6Hz,1H),6.28(d,J=7.6Hz,1H),5.59-5.54(m,1H),5.21(br s,1H),4.11(t,J=4.0Hz,2H),3.66(t,J=4.0Hz,2H),3.30(s,3H),3.28-3.25(m,1H),3.02-2.92(m,1H),2.24-2.16(m,1H),2.13-2.05(m,1H).MS(ESI)m/e[M+1]+ 407.9。 According to a similar preparation scheme for compound 9 (steps 7 and 8), from 7-(2-aminophenyl)-2-(4-hydroxyphenyl)-4,5,6,7-tetrahydropyrazolo[1 , 5-a]pyrimidine-3-carbonitrile and 2-bromoethyl methyl ether, the desired product is obtained under conditions well known to those skilled in the art. 1 H NMR (400MHz, DMSO- d6) δ 7.38 (d, J = 8.6 Hz, 2H), 7.00-6.94 (m, 3H), 6.75 (br s, 1H), 6.69 (d, J = 7.6Hz, 1H ), 6.51 (t, J = 7.6 Hz, 1H), 6.28 (d, J = 7.6 Hz, 1H), 5.59-5.54 (m, 1H), 5.21 (br s, 1H), 4.11 (t, J = 4.0) Hz, 2H), 3.66 (t, J = 4.0 Hz, 2H), 3.30 (s, 3H), 3.28-3.25 (m, 1H), 3.02-2.92 (m, 1H), 2.24-2.16 (m, 1H) , 2.13 - 2.05 (m, 1H). MS (ESI) m/e [M+1] + 407.9.
化合物22：7-(2-丙烯醯胺基苯基)-2-(4-(2-甲氧基乙氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 22 : 7-(2-Acrylaminophenyl)-2-(4-(2-methoxyethoxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1 ,5-a]pyrimidine-3-carboxamide
目標產物由化合物21和丙烯醯氯製備，與化合物8的步驟類似。1H NMR(400MHz,DMSO-d6)δ 9.84(s,1H),7.45(d,J=8.0Hz,1H),7.37(d,J=8.6,2H),7.30(t,J=7.6Hz,1H),7.21(t,J=7.6Hz,1H),6.93(d,J=8.6Hz,2H),6.81(s,1H),6.64(d,J=8.0Hz,1H),6.53(dd,J=10.5,17.0Hz,1H),6.27(dd,J=1.7,17.0Hz,1H),5.80-5.77(m,2H),4.11(t,J=4.4Hz,2H),3.66(t,J=4.4Hz,2H),3.30(s,3H),3.27-3.23(m,1H),3.03-2.94(m,1H),2.36-2.25(m,1H),2.01-1.95(m,1H).MS(ESI)m/e[M+1]+ 462.0。 The target product was prepared from compound 21 and acrylonitrile chloride, similar to the procedure for compound 8 . 1 H NMR (400MHz, DMSO- d 6) δ 9.84 (s, 1H), 7.45 (d, J = 8.0Hz, 1H), 7.37 (d, J = 8.6,2H), 7.30 (t, J = 7.6Hz , 1H), 7.21 (t, J = 7.6 Hz, 1H), 6.93 (d, J = 8.6 Hz, 2H), 6.81 (s, 1H), 6.64 (d, J = 8.0 Hz, 1H), 6.53 (dd , J =10.5, 17.0 Hz, 1H), 6.27 (dd, J = 1.7, 17.0 Hz, 1H), 5.80-5.77 (m, 2H), 4.11 (t, J = 4.4 Hz, 2H), 3.66 (t, J = 4.4 Hz, 2H), 3.30 (s, 3H), 3.27-3.23 (m, 1H), 3.03-2.94 (m, 1H), 2.36-2.25 (m, 1H), 2.01-1.95 (m, 1H) .MS (ESI) m/e [M+1] + 462.0.
實施例10：化合物23-24的合成Example 10: Synthesis of Compound 23-24
化合物23：7-(2-氨基苯基)-2-(4-(四氫-2H-吡喃-4-基氧)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 23 : 7-(2-Aminophenyl)-2-(4-(tetrahydro-2H-pyran-4-yloxy)phenyl)-4,5,6,7-tetrahydropyrazol[ 1,5-a]pyrimidine-3-carboxamide
步驟1：7-(2-氨基苯基)-2-(4-(四氫-2H-吡喃-4-基氧)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈 Step 1: 7-(2-Aminophenyl)-2-(4-(tetrahydro-2H-pyran-4-yloxy)phenyl)-4,5,6,7-tetrahydropyrazol[ 1,5-a]pyrimidine-3-carbonitrile
向7-(2-氨基苯基)-2-(4-羥基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈(33mg,0.1mmol)的THF(5mL)溶液中加入PPh3(78.6mg,0.25mmol)和四氫-2H-吡喃-4-醇(10mg,0.1mmol)。然後，氮氣保護0℃下，向混合物中逐滴加入DIAD(51mg,0.25mmol)，在0℃下攪拌10min。混合物允許升至室溫，並在室溫攪拌16小時。混合物減壓濃縮，加入到DCM(20mL)和飽和食鹽水(20mL)中。分離出有機層，硫酸鈉乾燥，製備矽膠板(DCM/CH3OH=10/1)純化，得到無色油狀物7-(2-氨基苯基)-2-(4-(四氫-2H-吡喃-4-基氧)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈(5mg,12%)。1H NMR(400MHz,DMSO-d6)δ 7.70(d,J=8.4Hz,2H),7.66-7.58(m,2H),7.58-7.52(m,1H), 7.03(d,J=8.4Hz,2H),6.97(t,J=7.6Hz,1H),6.69(d,J=8.0Hz,1H),6.49(t,J=7.6Hz,1H),6.23(d,J=8.0Hz,1H),5.63(s,1H),5.21(s,2H),4.66-4.56(m,1H),3.90-3.79(m,2H),3.54-3.43(m,2H),3.25-3.18(m,1H),2.97-2.86(m,1H),2.21-2.07(m,2H),2.02-1.92(m,2H),1.65-1.50(m,2H).MS(ESI)m/e[M+1]+ 415.9。 To 7-(2-aminophenyl)-2-(4-hydroxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carbonitrile (33 mg, PPh 3 (78.6 mg, 0.25 mmol) and tetrahydro-2H-pyran-4-ol (10 mg, 0.1 mmol) were added to a solution of EtOAc (5 mL). Then, under a nitrogen atmosphere at 0 ° C, DIAD (51 mg, 0.25 mmol) was added dropwise to the mixture and stirred at 0 ° C for 10 min. The mixture was allowed to warm to room temperature and stirred at room temperature for 16 hours. The mixture was concentrated under reduced pressure and added to DCM (20 mL) and brine. The organic layer was separated, dried over sodium sulfate, and then purified to silica gel (DCM/CH 3 OH = 10/1) to give 7-(2-aminophenyl)-2-(4-(tetrahydro-2H) -pyran-4-yloxy)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carbonitrile (5 mg, 12%). 1 H NMR (400MHz, DMSO- d 6) δ 7.70 (d, J = 8.4Hz, 2H), 7.66-7.58 (m, 2H), 7.58-7.52 (m, 1H), 7.03 (d, J = 8.4Hz , 2H), 6.97 (t, J = 7.6 Hz, 1H), 6.69 (d, J = 8.0 Hz, 1H), 6.49 (t, J = 7.6 Hz, 1H), 6.23 (d, J = 8.0 Hz, 1H) ), 5.63 (s, 1H), 5.21 (s, 2H), 4.66-4.56 (m, 1H), 3.90-3.79 (m, 2H), 3.54-3.43 (m, 2H), 3.25-3.18 (m, 1H) ), 2.97-2.86 (m, 1H), 2.21-2.07 (m, 2H), 2.02-1.92 (m, 2H), 1.65-1.50 (m, 2H). MS (ESI) m/e [M+1] + 415.9.
步驟2：7-(2-氨基苯基)-2-(4-(四氫-2H-吡喃-4-基氧)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 2: 7-(2-Aminophenyl)-2-(4-(tetrahydro-2H-pyran-4-yloxy)phenyl)-4,5,6,7-tetrahydropyrazol[ 1,5-a]pyrimidine-3-carboxamide
目標產物由7-(2-氨基苯基)-2-(4-(四氫-2H-吡喃-4-基氧基)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-腈製備，與化合物2的步驟2類似。1H NMR(400MHz,DMSO-d6)δ 7.38(d,J=8.8Hz,2H),7.02(d,J=8.8Hz,2H),6.95(t,J=7.6Hz,1H),6.75(s,1H),6.67(d,J=7.8Hz,1H),6.49(t,J=7.6Hz,1H),6.26(d,J=7.8Hz,1H),5.56(s,1H),5.16(s,2H),4.64-4.54(m,1H),3.91-3.79(m,2H),3.53-3.42(m,2H),3.29-3.19(m,1H),3.01-2.92(m,1H),2.25-2.05(m,2H),2.02-1.91(m,2H),1.64-1.52(m,2H).MS(ESI)m/e[M+1]+ 433.9。 The target product consists of 7-(2-aminophenyl)-2-(4-(tetrahydro-2H-pyran-4-yloxy)phenyl)-4,5,6,7-tetrahydropyrazole. [1,5-a]pyrimidine-3-carbonitrile was prepared in analogy to step 2 of compound 2 . 1 H NMR (400MHz, DMSO- d6) δ 7.38 (d, J = 8.8Hz, 2H), 7.02 (d, J = 8.8Hz, 2H), 6.95 (t, J = 7.6Hz, 1H), 6.75 (s , 1H), 6.67 (d, J = 7.8 Hz, 1H), 6.49 (t, J = 7.6 Hz, 1H), 6.26 (d, J = 7.8 Hz, 1H), 5.56 (s, 1H), 5.16 (s) , 2H), 4.64-4.54 (m, 1H), 3.91-3.79 (m, 2H), 3.53-3.42 (m, 2H), 3.29-3.19 (m, 1H), 3.01-2.92 (m, 1H), 2.25 -2.05 (m, 2H), 2.02-1.91 (m, 2H), 1.64-1.52 (m, 2H). MS (ESI) m/e [M+1] + 433.9.
化合物24：7-(2-丙烯醯胺基苯基)-2-(4-(四氫-2H-吡喃-4-基氧)苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 24 : 7-(2-Acrylaminophenyl)-2-(4-(tetrahydro-2H-pyran-4-yloxy)phenyl)-4,5,6,7-tetrahydropyridyl Oxazo[1,5-a]pyrimidine-3-carboxamide
目標產物由化合物23和丙烯醯氯製備，與化合物8的步驟類似。1H NMR(400MHz,DMSO-d6)δ 9.83(s,1H),7.45(d,J=8.0Hz,1H),7.38(d,J=8.6Hz,2H),7.30(t,J=7.6Hz,1H),7.21(t,J=7.6Hz,1H),7.02(d,J=8.6Hz,2H),6.81(s,1H),6.63(d,J=8.0Hz,1H),6.53(dd,J=17.0,10.3Hz,1H),6.27(dd,J=17.0,1.6Hz,1H),5.82-5.74(m,2H),4.66-4.51(m,1H),3.90-3.78(m,2H),3.54-3.40(m,2H),3.31-3.18(m,1H),3.03-2.93(m,1H),2.37-2.24(m,1H),2.04-1.92(m,3H),1.64-1.51(m,2H).MS(ESI)m/e[M+1]+ 487.9。 The target product was prepared from compound 23 and acrylonitrile chloride, similar to the procedure for compound 8 . 1 H NMR (400MHz, DMSO- d6) δ 9.83 (s, 1H), 7.45 (d, J = 8.0Hz, 1H), 7.38 (d, J = 8.6Hz, 2H), 7.30 (t, J = 7.6Hz , 1H), 7.21 (t, J = 7.6 Hz, 1H), 7.02 (d, J = 8.6 Hz, 2H), 6.81 (s, 1H), 6.63 (d, J = 8.0 Hz, 1H), 6.53 (dd , J =17.0, 10.3 Hz, 1H), 6.27 (dd, J = 17.0, 1.6 Hz, 1H), 5.82-5.74 (m, 2H), 4.66-4.51 (m, 1H), 3.90-3.78 (m, 2H) ), 3.54-3.40 (m, 2H), 3.31-3.18 (m, 1H), 3.03-2.93 (m, 1H), 2.37-2.24 (m, 1H), 2.04-1.92 (m, 3H), 1.64-1.51 (m, 2H). MS (ESI) m/e [M+1] + 487.9.
實施例11：化合物25-27的合成Example 11: Synthesis of Compound 25-27
化合物25：7-(1-(叔丁氧基羰基)哌啶-4-基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 25 : 7-(1-(tert-Butoxycarbonyl)piperidin-4-yl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1 ,5-a]pyrimidine-3-carboxamide
根據7-(3-氨基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈的步驟4到5和化合物2的步驟1到2的製備方案，在本領域普通技術人員熟知的條件下，由4-乙醯基哌啶-1-甲酸叔丁 酯和5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈，得到目標產物。1H NMR(CD3OD-d 4)δ 7.40(d,J=8.4Hz,2H),7.32-7.25(m,2H),7.06(t,J=7.6Hz,1H),7.01-6.94(m,4H),4.10-4.00(m,2H),3.98-3.91(m,1H),3.35-3.30(m,2H),2.70-2.58(m,2H),2.18-2.02(m,2H),2.02-1.84(m,1H),1.65-1.45(m,2H),1.39-1.12(m,2H),1.35(s,9H).MS(ESI)m/e[M+1]+ 518.0。 According to steps 4 to 5 of 7-(3-aminophenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile and step 1 of compound 2 Preparation of 2, tert-butyl 4-ethylhydrazine piperidine-1-carboxylate and 5-amino-3-(4-phenoxyphenyl)-1H- under conditions well known to those skilled in the art Pyrazole-4-carbonitrile gives the target product. 1 H NMR (CD 3 OD- d 4 ) δ 7.40 (d, J = 8.4 Hz, 2H), 7.32-7.25 (m, 2H), 7.06 (t, J = 7.6 Hz, 1H), 7.01-6.94 (m) , 4H), 4.10-4.00 (m, 2H), 3.98-3.91 (m, 1H), 3.35-3.30 (m, 2H), 2.70-2.58 (m, 2H), 2.18-2.02 (m, 2H), 2.02 - 1.84 (m, 1H), 1.65-1.45 (m, 2H), 1.39-1.12 (m, 2H), 1.35 (s, 9H). MS (ESI) m/e [M+1] + 518.0.
化合物26：7-(哌啶-4-基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺三氟乙酸鹽 Compound 26 : 7-(piperidin-4-yl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3- Formamide trifluoroacetate
目標產物由化合物25，根據化合物38的步驟2的類似方法合成。1H NMR(DMSO-d6)δ 8.47(br s,1H),8.16(br s,1H),7.50(d,J=8.4Hz,2H),7.46-7.38(m,2H),7.18(t,J=7.6Hz,1H),7.10-7.04(m,4H),6.72(br s,1H),4.08-4.01(m,1H),3.34-3.26(m,4H),2.94-2.75(m,2H),2.28-2.14(m,1H),2.07-1.88(m,2H),1.87-1.80(m,1H),1.75-1.66(m,1H),1.60-1.43(m,2H).MS(ESI)m/e[M+1]+ 418.0。 The title product was synthesized from compound 25 according to a procedure analogous to step 2 of compound 38 . 1 H NMR (DMSO-d 6 ) δ 8.47 (br s, 1H), 8.16 (br s, 1H), 7.50 (d, J = 8.4 Hz, 2H), 7.46-7.38 (m, 2H), 7.18 (t) , J = 7.6 Hz, 1H), 7.10-7.04 (m, 4H), 6.72 (br s, 1H), 4.08-4.01 (m, 1H), 3.34 - 3.26 (m, 4H), 2.94 - 2.75 (m, 2H), 2.28-2.14 (m, 1H), 2.07-1.88 (m, 2H), 1.87-1.80 (m, 1H), 1.75-1.66 (m, 1H), 1.60-1.43 (m, 2H). ESI) m/e [M+1] + 418.0.
化合物27：7-(1-丙烯醯基哌啶-4-基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 27 : 7-(1-Propylmercaptopiperidin-4-yl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a Pyrimidine-3-carboxamide
目標產物由化合物26和丙烯醯氯，根據化合物8的類似方法合成。1H NMR(DMSO-d6)δ 7.50(d,J=8.8Hz,2H),7.46-7.38(m,2H),7.17(t,J=7.6Hz,1H),7.08(d,J=7.6Hz,2H),7.05(d,J=8.8Hz,2H),6.83-6.76(m,1H),6.68(br s,1H),6.07(d,J=18.4Hz,1H),5.64(d,J=10.4Hz,1H),4.52-4.42(m,1H),4.11-3.98(m,2H),3.33-3.24(m,2H),3.04-2.94(m,1H),2.67-2.55(m,1H),2.33-2.25(m,1H),2.01-1.93(m,2H),1.78-1.66(m,1H),1.61-1.50(m,1H),1.30-1.18(m,2H).MS(ESI)m/e[M+1]+ 471.9. The title product was synthesized from Compound 26 and acryloyl chloride in a similar manner to Compound 8 . 1 H NMR (DMSO-d 6 ) δ 7.50 (d, J = 8.8 Hz, 2H), 7.46-7.38 (m, 2H), 7.17 (t, J = 7.6 Hz, 1H), 7.08 (d, J = 7.6) Hz, 2H), 7.05 (d, J = 8.8 Hz, 2H), 6.83-6.76 (m, 1H), 6.68 (br s, 1H), 6.07 (d, J = 18.4 Hz, 1H), 5.64 (d, J = 10.4 Hz, 1H), 4.52-4.42 (m, 1H), 4.11-3.98 (m, 2H), 3.33 - 3.24 (m, 2H), 3.04 - 2.94 (m, 1H), 2.67 - 2.55 (m, 1H), 2.33-2.25 (m, 1H), 2.01-1.93 (m, 2H), 1.78-1.66 (m, 1H), 1.61-1.50 (m, 1H), 1.30-1.18 (m, 2H). ESI)m/e[M+1] + 471.9.
通過手性製備高效液相色譜法，化合物27被分離成兩個對映立體異構體27a(峰1，R或S，在手性分析的保留時間為6.49min)，和27b(峰2，S或R，在手性分析的保留時間為8.03min)。手性分離條件如下所示：
手性分析條件如下所示：
實施例12：化合物28-29的合成Example 12: Synthesis of Compound 28-29
化合物28：7-(氮雜環丁烷-3-基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 28 : 7-(azetidin-3-yl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine 3-carbamamine
步驟1：3-(甲氧基(甲基)氨基甲醯基)氮雜環丁烷-1-甲酸叔丁酯 Step 1: 3-(Methoxy(methyl)carbamoyl)azetidin-1-carboxylic acid tert-butyl ester
向1-(叔丁氧基羰基)氮雜環丁烷-3-羧酸(5.15g,25.6mmol)的THF(100mL)溶液中，加入DCC(7.11g,34.5 mmol)，Et3N(5.18g,51.2mmol)和N,O-二甲基羥基胺鹽酸鹽(3.44g,35.3mmol)，將反應物在室溫下攪拌約16小時。減壓濃縮以除去溶劑，殘餘物在EA(100mL)和水(50mL)中分配，水相進一步用EA(50mL×3)萃取。合併有機相，飽和食鹽水(20mL)洗滌，減壓濃縮以除去溶劑，然後用矽膠層析柱(200-300目，CH2Cl2/MeOH=20/1)純化，得到無色油狀粗產物(~8.0g)。MS(ESI)m/e[M+23]+ 266.9,[M-55]+ 189.0。 A solution of 1- (t-butoxycarbonyl) azetidine-3-carboxylic acid (5.15g, 25.6mmol) in THF (100mL) were added DCC (7.11g, 34.5 mmol), Et 3 N (5.18 g, 51.2 mmol) and N , O -dimethylhydroxylamine hydrochloride (3.44 g, 35.3 mmol), and the mixture was stirred at room temperature for about 16 hours. The residue was taken up in EtOAc (EtOAc)EtOAc. Then (CH 2 Cl 2 / MeOH = 20/1 200-300 mesh,) was purified by column chromatography on silica gel combined organic phases with saturated brine (20mL), dried and concentrated under reduced pressure to remove the solvent, the crude product was obtained as a colorless oil (~8.0g). MS (ESI) m / e [ M + 23] + 266.9, [M-55] + 189.0.
步驟2：3-乙醯基氮雜環丁烷-1-甲酸叔丁酯 Step 2: 3-Ethyl azetidine-1-carboxylic acid tert-butyl ester
在0℃，向3-(甲氧基(甲基)氨基甲醯基)氮雜環丁烷-1-甲酸叔丁酯(7.0g,28.7mmol)的THF(150mL)溶液中，加入CH3MgBr(43mL,43mmol)，然後約2小時緩慢升至室溫。向混合物中加入10%檸檬酸水溶液(30mL)，並用EA(50mL×3)萃取，合併有機相，飽和食鹽水(20mL)洗滌，Na2SO4乾燥，過濾，濃縮，並用矽膠層析柱(200-300目，PE/EA=2/1)純化，得到無色油狀粗產物(4.0g,70%)。MS(ESI)m/e[M-55]+ 144.0。 At 0 deg.] C, a solution of 3- (methoxy (methyl) carbamoyl acyl) azetidine-1-carboxylate (7.0g, 28.7mmol) in THF (150mL) was added CH 3 MgBr (43 mL, 43 mmol) was then slowly warmed to room temperature over about 2 hours. A 10% aqueous citric acid solution (30 mL) was added to the mixture, and the mixture was extracted with EA (50 mL×3). The organic phase was combined, washed with saturated brine (20 mL), dried over Na 2 SO 4 , filtered, Purification of 200-300 mesh, PE / EA = 2 / 1) afforded crude product (4.0 g, 70%). MS (ESI) m/e [M-55] + 144.0.
化合物28：7-(氮雜環丁烷-3-基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 28 : 7-(azetidin-3-yl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine 3-carbamamine
根據7-(3-氨基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈的步驟4到5，化合物2的步驟1到2和化合物38的步驟2的製備方案，在本領域普通技術人員熟知的條件下，以3-乙醯基氮雜環丁烷-1-甲酸叔丁酯和5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈為起始原料，得到目標產物。1H NMR(DMSO-d 6)δ 8.70(br s,1H),8.44(br s,1H),7.50(d,J=8.6Hz,2H),7.45-7.40(m,2H),7.18(t,J=7.6Hz,1H),7.08(d,J=7.4Hz,2H),7.06(d,J=8.6Hz,2H),6.72(br s,1H),4.46-4.38(m,1H),4.24-4.15(m,1H),4.07-3.94(m,3H),3.29-3.24(m,2H),3.19-3.10(m,1H),2.13-2.04(m,1H),1.78-1.69(m,1H).MS(ESI)m/e[M+1]+ 390.0。 According to steps 4 to 5 of 7-(3-aminophenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile, step 1 of compound 2 is 2 and the preparation scheme of the step 2 of the compound 38 , under the conditions well known to those skilled in the art, tert-butyl 3-ethylhydrazinazetidine-1-carboxylate and 5-amino-3-(4- Phenoxyphenyl)-1H-pyrazole-4-carbonitrile is used as a starting material to give the desired product. 1 H NMR (DMSO- d 6 ) δ 8.70 (br s, 1H), 8.44 (br s, 1H), 7.50 (d, J = 8.6 Hz, 2H), 7.45-7.40 (m, 2H), 7.18 (t) , J = 7.6 Hz, 1H), 7.08 (d, J = 7.4 Hz, 2H), 7.06 (d, J = 8.6 Hz, 2H), 6.72 (br s, 1H), 4.46-4.38 (m, 1H), 4.24-4.15(m,1H), 4.07-3.94(m,3H), 3.29-3.24(m,2H), 3.19-3.10(m,1H),2.13-2.04(m,1H),1.78-1.69(m , 1H). MS (ESI) m/e [M+1] + 390.0.
化合物29：7-(1-丙烯醯基氮雜環丁烷-3-基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 29 : 7-(1-propenylfluorenylazetidin-3-yl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1, 5-a]pyrimidine-3-carboxamide
目標產物由化合物28和丙烯醯氯，根據化合物8的類似方法合成。1H NMR(DMSO-d 6)δ 7.50(d,J=8.4Hz,2H),7.44-7.40(m,2H),7.20-7.15(m,1H),7.10-7.04(m,4H),6.69(br s,1H),6.37-6.26(m,1H),6.12-6.04(m,1H),5.68-560(m,1H),4.43-4.25(m,2H),4.18-4.08(m,1H),4.04-3.96(m,1H),3.86-3.80(m,1H),3.32-3.26(m,2H),3.02-2.92(m,1H), 2.14-2.06(m,1H),1.79-1.70(m,1H).MS(ESI)m/e[M+1]+ 444.0。 The title product was synthesized from Compound 28 and acryloyl chloride in a similar manner to Compound 8 . 1 H NMR (DMSO- d 6 ) δ 7.50 (d, J = 8.4 Hz, 2H), 7.44-7.40 (m, 2H), 7.20-7.15 (m, 1H), 7.10-7.04 (m, 4H), 6.69 (br s, 1H), 6.37-6.26 (m, 1H), 6.12-6.04 (m, 1H), 5.68-560 (m, 1H), 4.43-4.25 (m, 2H), 4.18-4.08 (m, 1H) ), 4.04-3.96 (m, 1H), 3.86-3.80 (m, 1H), 3.32-3.26 (m, 2H), 3.02-2.92 (m, 1H), 2.14-2.06 (m, 1H), 1.79-1.70 (m, 1H). MS (ESI) m/e [M+1] + 444.0.
通過手性製備高效液相色譜法，化合物29被分離成兩個對映立體異構體29a(峰1，R或S，在手性分析的保留時間為10.54min)，和29b(峰2，S或R，在手性分析的保留時間為13.98min)。手性分離條件如下所示：
手性分析條件如下所示：
實施例13：化合物30-31的合成Example 13: Synthesis of Compound 30-31
化合物30和31：Cis-7-丙烯醯基-2-(4-苯氧基苯基)-4,5,5a,6,7,8,9,9a-八氫吡唑並[1,5-a]吡啶並[3,4-e]嘧啶-3-甲醯胺和Cis-7-丙烯醯基-2-(4-苯氧基苯基)-4,4a,5,6,7,8,8a,9-八氫吡唑並[1,5-a]吡啶並[4,3-d]嘧啶-3-甲醯胺 Compounds 30 and 31 : Cis-7-propenyl-2-(4-phenoxyphenyl)-4,5,5a,6,7,8,9,9a-octahydropyrazolo[1,5 -a] pyrido[3,4-e]pyrimidine-3-carboxamide and Cis-7-propenyl-2-(4-phenoxyphenyl)-4,4a,5,6,7, 8,8a,9-octahydropyrazolo[1,5-a]pyrido[4,3-d]pyrimidine-3-carboxamide
根據化合物27的製備方案，在本領域普通技術人員熟知的條件下，以4-氧代哌啶-1-甲酸叔丁酯和5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈為起始原料，得到目標產物。1H NMR(DMSO-d6)δ 7.51(d,J=8.4Hz,2H),7.46-7.38(m,2H),7.17(t,J=7.6Hz,1H),7.11-7.01(m,4H),6.90-6.78(m,1H),6.65(s,1H),6.18-6.06(m,1H),5.73-5.63(m,1H),4.45-4.33(m,1H),3.84-3.34(m,5H),3.22-3.16(m,1H),2.40-2.32(m,1H),2.21-2.10(m,1H),2.00-1.90(m,1H).MS(ESI)m/e[M+1]+ 443.9。 According to the preparation scheme of compound 27 , tert-butyl 4-oxopyridin-1-carboxylate and 5-amino-3-(4-phenoxyphenyl)-1H are used under conditions well known to those skilled in the art. Pyrazole-4-carbonitrile is used as a starting material to give the desired product. 1 H NMR (DMSO-d 6 ) δ 7.51 (d, J = 8.4 Hz, 2H), 7.46-7.38 (m, 2H), 7.17 (t, J = 7.6 Hz, 1H), 7.11-7.01 (m, 4H) ), 6.90-6.78 (m, 1H), 6.65 (s, 1H), 6.18-6.06 (m, 1H), 5.73-5.63 (m, 1H), 4.45-4.33 (m, 1H), 3.84-3.34 (m ,5H),3.22-3.16(m,1H), 2.40-2.32(m,1H),2.21-2.10(m,1H),2.00-1.90(m,1H).MS(ESI)m/e[M+ 1] + 443.9.
通過手性製備高效液相色譜法，化合物30被分離成兩個對映立體異構體30a(峰1，R或S，在手性分析的保留時間為
10.64min)，和30b(峰2，S或R，在手性分析的保留時間為15.18min)。手性分離條件如下所示：
手性分析條件如下所示：
化合物31是在製備化合物30過程中的一個副產物。1H NMR(DMSO-d6 at 80℃)δ 7.53(d,J=8.4Hz,2H),7.46- 7.38(m,2H),7.17(t,J=7.6Hz,1H),7.11-7.02(m,4H),6.82(dd,J=16.8,10.6Hz,1H),6.42(br s,1H),6.10(dd,J=16.8,2.3Hz,1H),6.01(br s,2H),5.68(dd,J=10.6,2.3Hz,1H),4.17(dd,J=5.4,12.2Hz,1H),3.67(t,J=12.2Hz,1H),3.28(td,J=4.0,10.4Hz,1H),2.24-2.17(m,1H),1.93-1.81(m,1H),1.47-1.33(m,1H).MS(ESI)m/e[M+1]+ 443.9。 Compound 31 is a by-product in the preparation of compound 30 . 1 H NMR (DMSO-d 6 at 80 ° C) δ 7.53 (d, J = 8.4 Hz, 2H), 7.46 - 7.38 (m, 2H), 7.17 (t, J = 7.6 Hz, 1H), 7.11 - 7.02 ( m, 4H), 6.82 (dd, J = 16.8, 10.6 Hz, 1H), 6.42 (br s, 1H), 6.10 (dd, J = 16.8, 2.3 Hz, 1H), 6.01 (br s, 2H), 5.68 (dd, J = 10.6, 2.3 Hz, 1H), 4.17 (dd, J = 5.4, 12.2 Hz, 1H), 3.67 (t, J = 12.2 Hz, 1H), 3.28 (td, J = 4.0, 10.4 Hz, 1H), 2.24 - 2.17 (m, 1H), 1.93-1.81 (m, 1H), 1.47-1.33 (m, 1H). MS (ESI) m/e [M+1] + 443.9.
實施例14：化合物32和33的合成Example 14: Synthesis of Compounds 32 and 33
化合物32：7-(氨基甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 32 : 7-(Aminomethyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide
步驟1：(E)-2-(4-(二甲胺基)-2-氧代-3-丁烯基)異吲哚啉-1,3-二酮 Step 1: ( E )-2-(4-(Dimethylamino)-2-oxo-3-butenyl)isoindoline-1,3-dione
N-丙酮基鄰苯二甲醯亞胺(1g,4.9mmol)溶於DMF-DMA(20mL)，加入一些4A的分子篩。反應混合物在氮氣保護下100℃攪拌15小時。冷卻至室溫，過濾，得到固體粗品(E)-2-(4-(二甲胺基)-2-氧代-3-丁烯基)異吲哚啉-1,3-二酮(600mg，47.5%)。MS(ESI)m/e[M+1]+ 259.1。 N -acetonyl phthalimide (1 g, 4.9 mmol) was dissolved in DMF-DMA (20 mL) and some 4A molecular sieves were added. The reaction mixture was stirred at 100 ° C for 15 hours under a nitrogen atmosphere. Cool to room temperature and filter to give crude ( E )-2-(4-(dimethylamino)-2-oxo-3-butenyl)isoindoline-1,3-dione (600 mg) , 47.5%). MS (ESI) m/e [M+1] + 259.1.
步驟2：7-((1,3-二氧代異吲哚啉-2-基)甲基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈 Step 2: 7-((1,3-Dioxaisoindolin-2-yl)methyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine- 3-carbonitrile
將(E)-2-(4-(二甲胺基)-2-氧代-3-丁烯基)異吲哚啉-1,3-二酮(600mg,2.33mmol)和5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈(642mg,2.33mmol)加入到醋酸中(20mL)。混合物加熱到120℃，攪拌15小時。濃縮，殘餘物懸浮在30mL溶劑(PE/EA=4/1)。過濾，固體通過製備矽膠板純化(DCM/EA=50/1)，得到固體7-((1,3-二氧代異吲哚啉-2-基)甲基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈(430mg,40%)。1H NMR(DMSO-d6)δ 8.74(d,J=4.4Hz,1H),8.05(d,J=8.8Hz,2H),,7.96-7.90(m,1H),7.88-7.85(m,2H),7.5(d,J=4.4Hz,1H),7.46-7.37(m,2H),7.21-7.09(m,5H),5.32(s,2H).MS(ESI)m/e[M+1]+ 472.1。 ( E )-2-(4-(Dimethylamino)-2-oxo-3-butenyl)isoindoline-1,3-dione (600 mg, 2.33 mmol) and 5-amino- 3-(4-Phenoxyphenyl)-1H-pyrazole-4-carbonitrile (642 mg, 2.33 mmol) was added toEtOAc (20 mL). The mixture was heated to 120 ° C and stirred for 15 hours. Concentrate and the residue was suspended in 30 mL of solvent (PE/EA = 4/1). Filtration, the solid was purified by preparative silica gel (DCM / EA = 50/1) to give solid 7-((1,3-dioxoisoindolin-2-yl)methyl)-2-(4-benzene Oxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile (430 mg, 40%). 1 H NMR (DMSO-d 6 ) δ 8.74 (d, J = 4.4 Hz, 1H), 8.05 (d, J = 8.8 Hz, 2H), 7.96-7.90 (m, 1H), 7.88-7.85 (m, 2H), 7.5 (d, J = 4.4 Hz, 1H), 7.46-7.37 (m, 2H), 7.21-7.09 (m, 5H), 5.32 (s, 2H). MS (ESI) m/e [M+ 1] + 472.1.
步驟3：7-(氨基甲基)-2-(4-苯氧基苯基)吡唑[1,5-a]嘧啶-3-甲腈 Step 3: 7-(Aminomethyl)-2-(4-phenoxyphenyl)pyrazole [1,5-a]pyrimidine-3-carbonitrile
7-((1,3-二氧代異吲哚啉-2-基)甲基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈(290mg,0.62mmol)溶於甲醇(5mL)和1,4-二氧六環(5mL)，加入水合肼(12滴)。將反應混合物 加熱至60℃攪拌3小時。將該混合物濃縮，懸浮在20mL溶劑(DCM/MeOH=10/1)。過濾，濃縮濾液，快速色譜柱純化(淋洗劑先用乙酸乙酯，再用DCM/MeOH=10/1)得到固體7-(氨基甲基)-2-(4-苯氧基苯基)吡唑[1,5-a]嘧啶-3-甲腈(150mg,71%)。 MS(ESI)m/e[M+1]+ 342.1。 7-((1,3-Dioxaisoindolin-2-yl)methyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-A The nitrile (290 mg, 0.62 mmol) was dissolved in methanol (5 mL) and 1,4-dioxane (5 mL). The reaction mixture was heated to 60 ° C and stirred for 3 hours. The mixture was concentrated and suspended in 20 mL of solvent (DCM /MeOH = 10/1). Filtration, concentrating the filtrate, purification by flash column (eluent with ethyl acetate and then DCM /MeOH = 10/1) to give solid 7-(aminomethyl)-2-(4-phenoxyphenyl) Pyrazole [1,5-a]pyrimidine-3-carbonitrile (150 mg, 71%). MS (ESI) m/e [M+1] + 3421.
步驟4：7-(氨基甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈 Step 4: 7-(Aminomethyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carbonitrile
7-(氨基甲基)-2-(4-苯氧基苯基)吡唑[1,5-a]嘧啶-3-甲腈(150mg,0.44mmol)溶於乙醇(20mL)，加入NaBH4(200mg)。將該反應混合物在室溫下攪拌15小時。將該混合物濃縮除去溶劑，向殘餘物中加入乙酸乙酯(20mL)和水(20mL)。有機相濃縮，得到固體粗品7-(氨基甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈(150mg,100%)，將其用於下一步驟未經進一步純化。MS(ESI)m/e[M+1]+ 346.0。 7-(Aminomethyl)-2-(4-phenoxyphenyl)pyrazole [1,5-a]pyrimidine-3-carbonitrile (150 mg, 0.44 mmol) was dissolved in ethanol (20 mL) and NaBH 4 (200mg). The reaction mixture was stirred at room temperature for 15 hours. The mixture was concentrated to remove solvent and ethyl acetate (20mL) and water (20mL). The organic phase was concentrated to give crude solid 7-(aminomethyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3 -Methanol (150 mg, 100%) which was used in the next step without further purification. MS (ESI) m / e [ M + 1] + 346.0.
步驟5：7-(氨基甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 5: 7-(Aminomethyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide
向7-(氨基甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈(15mg,0.043mmol)的EtOH(2mL)溶液中，加入DMSO(1mL)，的NaOH(5N,0.5mL)和的過氧化氫(30%水溶液，0.5mL)。在60℃下攪拌2小時後，將混合物濃縮，除去乙醇。將殘餘物在水(30mL)和EA(20mL)之間分配。將有機相濃縮，製備HPLC純化，水相含0.1% TFA，20%到40% CH3CN梯度洗脫，收集合併目標產品，過夜凍幹，得到產物的TFA鹽(10mg,64%)。1H NMR(400MHz,DMSO-d6)δ 8.07(br s,3H),7.62(d,J=8.8Hz,2H),7.43-7.35(m,2H),7.25(t,J=7.6Hz,1H),7.15-7.12(m,4H),6.77(br s,1H),4.38-4.30(m,1H),3.40-3.20(m,4H),2.16-2.06(m,1H),2.00-1.80(m,1H).MS(ESI)m/e[M+1]+ 364.0。 To 7-(aminomethyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carbonitrile (15 mg, To a solution of 0.043 mmol) in EtOH (2 mL), EtOAc (1 mL), EtOAc (5N, 0.5mL) After stirring at 60 ° C for 2 hours, the mixture was concentrated to remove ethanol. The residue was partitioned between water (30 mL) and EA (20 mL). The organic phase was concentrated and purified by preparative HPLC, an aqueous phase containing 0.1% TFA, 20% to 40% CH 3 CN gradient, to collect the target product were combined, lyophilized overnight to give the product as a TFA salt (10mg, 64%). 1 H NMR (400MHz, DMSO- d 6) δ 8.07 (br s, 3H), 7.62 (d, J = 8.8Hz, 2H), 7.43-7.35 (m, 2H), 7.25 (t, J = 7.6Hz, 1H), 7.15-7.12 (m, 4H), 6.77 (br s, 1H), 4.38-4.30 (m, 1H), 3.40-3.20 (m, 4H), 2.16-2.06 (m, 1H), 2.00-1.80 (m, 1H). MS (ESI) m/e [M+1] + 364.0.
化合物33：7-(丙烯醯氨基甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 33: 7-(Protonium aminomethyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-A Guanamine
向7-(氨基甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺(125mg,0.344mmol)的DCM(5mL)溶液中，加入三乙胺(4滴)和丙烯醯氯(46.5mg,0.52mmol)。在室溫下攪拌30分鐘後，混合物在水(10mL)和DCM(5mL)中分配。將有機層濃縮，製備TLC(DCM/MeOH=20/1)純化，得到產品(40mg,28%)。1H NMR(400MHz,DMSO-d6)δ 8.37(t,J=6.0Hz,1H),7.59(d,J=8.8Hz,2H),7.54-7.43(m,2H),7.25(t,J=7.6Hz,1H),7.16-7.11(m,4H),6.76(s,1H),6.33(dd,J=10.1,17.0Hz,1H),6.18(dd,J=1.9,17.0Hz,1H),5.69(dd,J=1.9,10.1Hz,1H),4.28-4.22(m,1H),3.92-3.80(m,1H),3.50-3.30(m,3H),2.14-1.94(m,2H).MS(ESI)m/e[M+1]+ 417.9。 To 7-(aminomethyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide (125 mg To a solution of 0.34 mmol of DCM (5 mL), triethylamine (4 drops) and EtOAc (46.5 mg, 0.52 mmol). After stirring at room temperature for 30 minutes, the mixture was partitioned w~~~~ The organic layer was concentrated to give EtOAc (EtOAc:EtOAc) 1 H NMR (400MHz, DMSO- d 6) δ 8.37 (t, J = 6.0Hz, 1H), 7.59 (d, J = 8.8Hz, 2H), 7.54-7.43 (m, 2H), 7.25 (t, J = 7.6 Hz, 1H), 7.16-7.11 (m, 4H), 6.76 (s, 1H), 6.33 (dd, J = 10.1, 17.0 Hz, 1H), 6.18 (dd, J = 1.9, 17.0 Hz, 1H) , 5.69 (dd, J = 1.9, 10.1 Hz, 1H), 4.28-4.22 (m, 1H), 3.92-3.80 (m, 1H), 3.50-3.30 (m, 3H), 2.14-1.94 (m, 2H) .MS (ESI) m/e [M+1] + 417.9.
通過手性製備高效液相色譜法，化合物33被分離成兩個對映立體異構體33a(峰1，R或S，在手性分析的保留時間為6.04min)，和33b(峰2，S或R，在手性分析的保留時間為8.87min)。手性分離條件如下所示：
手性分析條件如下所示：
實施例15：化合物34和35的合成Example 15: Synthesis of Compounds 34 and 35
化合物34：7-((甲氨基)甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 34 : 7-((Methylamino)methyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3- Formamide
步驟1：N-((3-氰基-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-7-基)甲基)-2,2,2-三氟乙醯胺 Step 1: N -((3-Cyano-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidin-7-yl)methyl)-2,2,2-tri Fluoroacetamide
向7-(氨基甲基)-2-(4-苯氧基苯基)吡唑[1,5-a]嘧啶-3-甲腈(50mg,0.147mmol)的5mL DCM溶液中加入DIEA(3滴)和Tf-2O(2滴)。反應混合物在室溫攪拌2小時，然後加入水(10mL)，DCM(5mL×2)萃取。DCM層旋幹，得到黃色固體產品(50mg,78%)，未經進一步純化直接用於下步反應。 Add DIEA (3) to a solution of 7-(aminomethyl)-2-(4-phenoxyphenyl)pyrazole [1,5-a]pyrimidine-3-carbonitrile (50 mg, 0.147 mmol) in 5 mL DCM Drop) and Tf- 2 O (2 drops). The reaction mixture was stirred at room temperature for 2 hr then water (10 mL) was evaporated. The DCM layer was dried <RTI ID=0.0></RTI> to <RTI ID=0.0>
步驟2：N-((3-氰基-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-7-基)甲基)-2,2,2-三氟-N-甲基乙醯胺 Step 2: N -((3-Cyano-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidin-7-yl)methyl)-2,2,2-tri Fluorine- N -methylacetamide
向N-((3-氰基-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-7-基)甲基)-2,2,2-三氟乙醯胺(40mg,0.091mmol)的丙酮(5mL)溶液中，加入K2CO3(50mg)和CH3I(0.5mL)。室溫攪拌4小時後，濃縮混合物。殘餘物加入到水(10mL)和DCM(10mL)中。有機層濃縮，然後用製備矽膠板(DCM/MeOH=20/1)純化，得到黃色固體(30mg,73%)。MS(ESI)m/e[M+1]+ 451.9。 To N -((3-cyano-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidin-7-yl)methyl)-2,2,2-trifluoroethyl Amides (40mg, 0.091mmol) in acetone (5mL) was added K 2 CO 3 (50mg) and CH 3 I (0.5mL). After stirring at room temperature for 4 hours, the mixture was concentrated. The residue was taken into water (10 mL) and DCM (10 mL). The organic layer was concentrated and purified with EtOAc EtOAc EtOAc EtOAc MS (ESI) m/e [M+1] + 451.9.
步驟3：7-((甲氨基)甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈 Step 3: 7-((Methylamino)methyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3- Nitrile
向N-((3-氰基-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-7-基)甲基)-2,2,2-三氟-N-甲基乙醯胺(40mg,0.089mmol)的EtOH(15mL)溶液中，加入NaBH4(50mg)。室溫攪拌30min後，混合物濃縮。殘餘物加入到水(20mL)和EA(20mL)中。有機層濃縮，製備矽膠板(DCM/MeOH=5/1)純化，得到白色固體(20mg,63%)。MS(ESI)m/e[M+1]+ 359.9。 To N -((3-cyano-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidin-7-yl)methyl)-2,2,2-trifluoro- N - methyl as acetamide (40mg, 0.089mmol) in EtOH (15mL) was added NaBH 4 (50mg). After stirring at room temperature for 30 min, the mixture was concentrated. The residue was taken up in water (20 mL) and EA (20 mL). The organic layer was concentrated and purified EtOAc EtOAcjjjjjj MS (ESI) m/e [M+1] + 359.9.
步驟4：7-((甲氨基)甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 4: 7-((Methylamino)methyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3- Formamide
向7-((甲氨基)甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈(20mg,0.0557mmol)的EtOH(2mL)溶液中，依次加入DMSO(1mL),NaOH(5N,0.5mL)和H2O2(30%溶液，0.5mL)。反應混合物60℃攪拌1小時，混合物濃縮除去EtOH。殘餘物加入到在水(20mL)和EA(20mL)中。EA層濃縮，得到白色固體產品(10mg,48%)。1H NMR(400MHz,DMSO-d6)δ 7.51(d,J=8Hz,2H),7,47-7.40(dm,2H),7.19 (t,J=7.6Hz,1H),7.11-7.04(m,4H),6.61(s,1H),4.26-4.16(m,1H),3.34-3.27(m,2H),2.97-2.95(m,1H),2.83-2.77(m,1H),2.33(s,3H),2.08-2.02(m,2H).MS(ESI)m/e[M+1]+ 378.0。 To 7-((methylamino)methyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carbonitrile (20mg, 0.0557mmol) in EtOH (2mL) solution was added sequentially DMSO (1mL), NaOH (5N , 0.5mL) , and H 2 O 2 (30% solution, 0.5mL). The reaction mixture was stirred at 60 ° C for 1 hour, and the mixture was concentrated to remove EtOH. The residue was taken up in water (20 mL) and EA (20 mL). The EA layer was concentrated to give a white solid product (10 mg, 48%). 1 H NMR (400 MHz, DMSO-d6) δ 7.51 (d, J = 8 Hz, 2H), 7, 47-7.40 (dm, 2H), 7.19 (t, J = 7.6 Hz, 1H), 7.11 - 7.04 (m) , 4H), 6.61(s,1H), 4.26-4.16(m,1H),3.34-3.27(m,2H),2.97-2.95(m,1H),2.83-2.77(m,1H),2.33(s , 3H), 2.08-2.02 (m, 2H). MS (ESI) m/e [M+1] + 378.0.
化合物35：7-((N-甲基(丙烯醯胺基)甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 35 : 7-(( N -methyl(acrylamido)methyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5 -a]pyrimidine-3-carboxamide
向7-((甲氨基)甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺(92mg,0.244mmol)的DCM(5mL)溶液中，加入Et3N(3滴)和丙烯醯氯(33mg,0.366mmol)。反應混合物室溫攪拌30min。然後，將混合物在水(30mL)和DCM(20mL)中分配。將有機層濃縮，製備矽膠板(DCM/MeOH=15/1)純化，得到白色固體產品(25mg,24%)。1H NMR(400MHz,DMSO-d6和D2O at 80℃)δ 7.51(d,J=8.4Hz,2H),7.44-7.39(m,2H),7.20-7.14(m,1H),7.10-7.04(m,4H),6.75-6.57(m,2H),6.10-5.85(m,1H),5.67-5.50(m,1H),4.45-4.38(m,1H),4.00-3.70(m,2H),3.40-3.30(m,2H),3.00(s,3H),2.14-1.90(m,2H).MS(ESI)m/e[M+1]+ 432.0。 To 7-((methylamino)methyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamidine amine (92mg, 0.244mmol) in DCM (5mL) was added Et 3 N (3 drops) and Bing Xixi chloride (33mg, 0.366mmol). The reaction mixture was stirred at room temperature for 30 min. The mixture was then partitioned between water (30 mL) and DCM (20 mL). The organic layer was concentrated to give EtOAc (EtOAc:EtOAc) 1 H NMR (400 MHz, DMSO-d 6 and D 2 O at 80 ° C) δ 7.51 (d, J = 8.4 Hz, 2H), 7.44-7.39 (m, 2H), 7.20-7.14 (m, 1H), 7.10 -7.04 (m, 4H), 6.75-6.57 (m, 2H), 6.10-5.85 (m, 1H), 5.67-5.50 (m, 1H), 4.45-4.38 (m, 1H), 4.00-3.70 (m, 2H), 3.40-3.30 (m, 2H), 3.00 (s, 3H), 2.14-1.90 (m, 2H). MS (ESI) m/e [M+1] + 432.0.
實施例16：化合物36和37的合成Example 16: Synthesis of Compounds 36 and 37
化合物36：7-(氨基甲基)-2-(1-苄基-1H-吡唑-4-基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺三氟乙酸鹽 Compound 36: 7- (aminomethyl) -2- (1-benzyl -1H- pyrazol-4-yl) -4,5,6,7-tetrahydro-pyrazolo [1,5-a] pyrimidine 3-carbamidine trifluoroacetate
步驟1：1-苄基吡唑-4-甲酸乙酯 Step 1: 1-Benzylpyrazole-4-carboxylic acid ethyl ester
向1H-吡唑-4-羧酸乙酯(35.0g,250mmol)和K2CO3(69.0g,500mmol)在CH3CN(250mL)中的混合物，加入BnBr(42.7g,250mmol)。混合物室溫攪拌18小時，濃縮。將殘餘物懸浮在EA(500mL)中，用水(200mL×2)洗滌，Na2SO4乾燥，濃縮，得到白色固體目標產物(53.0g,91.2%)。1H NMR(DMSO-d6)δ 8.46(s,1H),7.88(s,1H),7.41-7.19(m,5H),5.37(s,2H),4.21(q,2H, J =5.4Hz),1.25(t,3H, J =5.4Hz).MS(ESI)m/e[M+1]+ 231.0。 To 1H- pyrazole-4-carboxylate (35.0g, 250mmol), and K in CH 3 CN (250mL) mixture of 2 CO 3 (69.0g, 500mmol) , was added BnBr (42.7g, 250mmol). The mixture was stirred at room temperature for 18 hours and concentrated. The residue was suspended in EA (500mL), washed, Na dried washed with water (200mL × 2) 2 SO 4 , and concentrated to give the desired product as a white solid (53.0g, 91.2%). 1 H NMR (DMSO-d 6 ) δ 8.46 (s, 1H), 7.88 (s, 1H), 7.41-7.19 (m, 5H), 5.37 (s, 2H), 4.21. (q, 2H, J = 5.4 Hz ), 1.25 (t, 3H, J = 5.4 Hz). MS (ESI) m/e [M+1] + 231.0.
步驟2：1-苄基-1H-吡唑-4-羧酸 Step 2: 1-Benzyl-1H-pyrazole-4-carboxylic acid
1-苄基吡唑-4-甲酸乙酯(53.0g,0.23mol)和LiOH(19.4g,0.46mol)在THF(100mL)和水(100mL)中的混合 物，回流攪拌6小時。然後，蒸去THF，殘餘物由6N鹽酸酸化，沉澱形成，過濾，乾燥，得到白色固體目標產物(44.0g,92.8%)。1H NMR(DMSO-d6)δ 12.36(s,1H),8.39(s,1H),7.85(s,1H),7.38-7.27(m,5H),5.37(s,2H).MS(ESI)m/e[M+1]+ 202.9。 A mixture of ethyl 1-benzylpyrazole-4-carboxylate (53.0 g, 0.23 mol) and EtOAc (1. After the THF was evaporated, the residue was crystalljjjjjjjjjjjjjjjj 1 H NMR (DMSO-d6) δ 12.36 (s, 1H), 8.39 (s, 1H), 7.85 (s, 1H), 7.38-7.27 (m, 5H), 5.37 (s, 2H). MS (ESI) m/e [M+1] + 202.9.
步驟3：2-((1-苄基-1H-吡唑-4-基)(羥基)亞甲基)丙二腈 Step 3: 2-((1-Benzyl-1H-pyrazol-4-yl)(hydroxy)methylene)malononitrile
1-苄基-1H-吡唑-4-羧酸(25.0g,123.8mmol)的SOCl2(250mL)溶液加熱至回流3小時。該混合物濃縮，得到中間體，無需進一步純化直接用於下一步反應。在0-5℃，向丙二腈(8.2g,12.8mmol)和DIEA(32.0g,247.6mmol)的THF(250mL)的溶液中，緩慢滴加中間體的甲苯(250mL)溶液，約1小時滴畢。混合物室溫攪拌16小時後，用水(500mL)淬滅，EA(500mL×3)萃取。合併有機相，分別用3N HCl(500mL)，飽和食鹽水(500mL×3)洗滌，Na2SO4乾燥，濃縮得到黃色固體產物(26.5g,85.0%)。MS(ESI)m/e[M+1]+ 250.9。 1-benzyl--1H- pyrazole-4-carboxylic acid (25.0g, 123.8mmol) in SOCl 2 (250mL) was heated to reflux for 3 hours. The mixture was concentrated to give an intermediate which was used in the next step without further purification. A solution of the intermediate in toluene (250 mL) was slowly added dropwise to a solution of malononitrile (8.2 g, 12.8 mmol) and DIEA (32.0 g, 247.6 mmol) in THF (250 mL). Dropped. The mixture was stirred at room temperature for 16 h then EtOAc (EtOAc) The combined organic phases were washed, dried Na 2 SO 4, and concentrated to give a yellow solid (26.5g, 85.0%) with 3N HCl (500mL), saturated brine (500mL × 3). MS (ESI) m / e [ M + 1] + 250.9.
步驟4：2-((1-苄基-1H-吡唑-4-基)(甲氧基)亞甲基)丙二腈 Step 4: 2-((1-Benzyl-1H-pyrazol-4-yl)(methoxy)methylene)malononitrile
2-((1-苄基-1H-吡唑-4-基)(羥基)亞甲基)丙二腈(26.5g,106mmol)的CH(OMe)3(250mL)，加熱至75℃，攪拌16小時。混合物濃縮，殘餘物用MeOH(50mL)洗滌，得到類白色固體2-((1-苄基-1H-吡唑-4-基)(甲氧基)亞甲基)丙二腈(14.5g，51.8%)。1H NMR(DMSO-d6)δ 8.71(s,1H),8.08(s,1H),7.42-7.24(m,5H),5.46(s,2H),4.12(s,3H).MS(ESI)m/e[M+1]+ 264.9。 2-((1-Benzyl-1H-pyrazol-4-yl)(hydroxy)methylene)malononitrile (26.5 g, 106 mmol) in CH (OMe) 3 (250 mL), warmed to 75 ° C, stirred 16 hours. The mixture was concentrated, and the~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 51.8%). 1 H NMR (DMSO-d 6 ) δ 8.71 (s, 1H), 8.08 (s, 1H), 7.42 - 7.24 (m, 5H), 5.46 (s, 2H), 4.12 (s, 3H). )m/e[M+1] + 264.9.
步驟5：5-氨基-2'-苄基-3,4'-雙(1H-吡唑)-4-甲腈 Step 5: 5-Amino-2'-benzyl-3,4'-bis(1H-pyrazole)-4-carbonitrile
2-((1-苄基-1H-吡唑-4-基)(甲氧基)亞甲基)丙二腈(14.5g,54.9mmol)和水合肼(10mL)在乙醇(500mL)中的混合物，室溫攪拌4小時。混合物濃縮得到粗產物，MeOH洗滌，得到類白色固體產品5-氨基-2'-苄基-3,4'-雙(1H-吡唑)-4-甲腈(10g,69.0%)。1H NMR(DMSO-d6)δ 11.76(br s,1H),8.18(s,1H),7.82(s,1H),7.34-7.26(m,5H),6.11(br s,2H),5.40(s,2H).MS(ESI)m/e[M+1]+ 264.9。 2-((1-Benzyl-1H-pyrazol-4-yl)(methoxy)methylene)malononitrile (14.5 g, 54.9 mmol) and hydrazine hydrate (10 mL) in ethanol (500 mL) The mixture was stirred at room temperature for 4 hours. The mixture was concentrated to give abr. 1 H NMR (DMSO-d 6 ) δ 11.76 (br s, 1H), 8.18 (s, 1H), 7.82 (s, 1H), 7.34-7.26 (m, 5H), 6.11 (br s, 2H), 5.40 (s, 2H). MS (ESI) m/e [M+1] + 264.9.
化合物36：7-(氨基甲基)-2-(1-苄基-1H-吡唑-4-基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺三氟乙酸鹽 Compound 36 : 7-(Aminomethyl)-2-(1-benzyl-1H-pyrazol-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine 3-carbamidine trifluoroacetate
根據化合物32的步驟2到5的類似方法，在本領域普通技術人員熟知的條件下，以5-氨基-2'-苄基-3,4'-雙(1H-吡唑)-4-甲腈和(E)-2-(4-(二甲胺基)-2-氧代-3-丁烯基)異吲哚啉-1,3-二酮為起始原料，得到目標產物。1H NMR(400MHz,DMSO-d6)δ 8.14(s,1H),7.97(br s,3H),7.74(s,1H),739-7.24(m,5H),5.37(s,2H),4.40-4.25(m,1H),3.37-3.16(m,4H),2.16-2.08(m,1H),1.99-1.89(m,1H).MS(ESI)m/e[M+1]+ 352.0。 According to a similar method compound 32 step 2 to 5, under conditions well known to those of ordinary skill in the art, the 5-amino-2'-benzyloxy-3,4'-bis (lH-pyrazol) -4- The nitrile and ( E )-2-(4-(dimethylamino)-2-oxo-3-butenyl)isoindoline-1,3-dione are used as starting materials to give the desired product. 1 H NMR (400MHz, DMSO- d 6) δ 8.14 (s, 1H), 7.97 (br s, 3H), 7.74 (s, 1H), 739-7.24 (m, 5H), 5.37 (s, 2H), 4.40-4.25(m,1H), 3.37-3.16(m,4H), 2.16-2.08(m,1H), 1.99-1.89(m,1H).MS(ESI)m/e[M+1] + 352.0 .
化合物37：7-(丙烯醯氨基甲基)-2-(1-苄基-1H-吡唑-4-基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 37 : 7-(Acrylaminomethyl)-2-(1-benzyl-1H-pyrazol-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a Pyrimidine-3-carboxamide
目標產物由化合物36和丙烯醯氯，根據化合物8的類似方法合成。1H NMR(400MHz,DMSO-d6)δ 8.27(t,J=6.1Hz,1H),8.11(s,1H),7.67(s,1H),7.41-7.21(m,5H),6.58(s,1H),6.25(br s,2H),6.25(dd,J=17.1,10.1Hz,1H),6.10(dd,J=17.1,2.1Hz,1H),5.62(dd,J=10.1,2.1Hz,1H),5.36(s,2H),4.16-4.10(m,1H),3.83-3.72(m,1H),3.42-3.30(m,1H), 3.30-3.22(m,2H),2.00-1.96(m,1H),1.95-1.86(m,1H).MS(ESI)m/e[M+1]+ 405.9。 The target product was synthesized from Compound 36 and acryloyl chloride in a similar manner to Compound 8 . 1 H NMR (400MHz, DMSO- d 6) δ 8.27 (t, J = 6.1Hz, 1H), 8.11 (s, 1H), 7.67 (s, 1H), 7.41-7.21 (m, 5H), 6.58 (s , 1H), 6.25 (br s, 2H), 6.25 (dd, J = 17.1, 10.1 Hz, 1H), 6.10 (dd, J = 17.1, 2.1 Hz, 1H), 5.62 (dd, J = 10.1, 2.1 Hz ,1H), 5.36(s,2H), 4.16-4.10(m,1H),3.83-3.72(m,1H),3.42-3.30(m,1H), 3.30-3.22(m,2H),2.00-1.96 (m, 1H), 1.95-1.86 (m, 1H). MS (ESI) m/e [M+1] + 405.9.
實施例17：化合物38到40的合成Example 17: Synthesis of Compounds 38 to 40
化合物38：1'-苄基-6-(4-苯氧基苯基)-1,2-二氫螺[咪唑[1,2-b]吡唑-3,4'-哌啶]-7-甲醯胺 Compound 38 : 1'-Benzyl-6-(4-phenoxyphenyl)-1,2-dihydrospiro[imidazole[1,2-b]pyrazole-3,4'-piperidine]-7 -Procarbamide
步驟1：1-(1-苄基-4-(乙氧羰基)哌啶-4)肼基-1,2-二羧酸二叔丁酯 Step 1: 1-(1-Benzyl-4-(ethoxycarbonyl)piperidin-4)nonyl-1,2-dicarboxylic acid di-tert-butyl ester
在氮氣保護下，控溫-75℃，向二異丙胺(153mg,1.5mmol)的THF(20mL)溶液中緩慢滴加n-BuLi(2.5M,0.6mL)。5min後，加入1-苄基-4-哌啶甲酸乙酯(247mg,1.0mmol)。將混合物在-70℃下攪拌10min，然後再加入偶氮二甲酸二叔丁酯(345mg,1.5mmol)，繼續攪拌30min。NH4Cl水溶液(10mL)淬滅反應，然後用乙酸乙酯(10mL×3)萃取。合併有機相，Na2SO4乾燥，濃縮，得到類白色固體粗產品(350mg,72%)。MS(ESI,m/e)[M+1]+ 478.3。 n-BuLi (2.5 M, 0.6 mL) was slowly added dropwise to a solution of diisopropylamine (153 mg, 1.5 mmol) in THF (20 mL). After 5 min, ethyl 1-benzyl-4-piperidinecarboxylate (247 mg, 1.0 mmol) was added. The mixture was stirred at -70 <0>C for 10 min then additional di-tert-butyl azodicarboxylate (345 mg, 1.5 mmol) was added and stirring was continued for 30 min. NH 4 Cl aq (10 mL) quenched the reaction and then extracted with ethyl acetate (10mL × 3). The organic phases were combined, Na dried 2 SO 4, and concentrated to give an off-white solid crude product (350mg, 72%). MS (ESI, m/e) [M+1] + 478.3.
步驟2：1-苄基-4-肼基-4'-甲酸乙酯基哌啶鹽酸鹽 Step 2: 1-Benzyl-4-indolyl-4'-ethyl ester piperidine hydrochloride
1-(1-苄基-4-(乙氧羰基)哌啶-4)肼基-1,2-二羧酸二叔丁酯(1.0g,2.09mmol)和con.HCl(1.0mL)的MeOH(10mL)溶液，加熱回流2小時。混合物濃縮，得到黃色固體的目標產物(650mg,88.9%)。MS(ESI,m/e)[M+1]+ 278.0. 1-(1-Benzyl-4-(ethoxycarbonyl)piperidin-4)nonyl-1,2-dicarboxylic acid di-tert-butyl ester (1.0 g, 2.09 mmol) and con.HCl (1.0 mL) A solution of MeOH (10 mL) was heated and reflux for 2 h. The mixture was concentrated to give the title compound ( 650 mg, 88. MS (ESI, m/e) [M+1] + 278.0.
步驟3：4-(5-氨基-4-氰基-3-(4-苯氧基苯基)-1H-吡唑-1-基)-1-苄基哌啶-4-羧酸乙酯 Step 3: 4-(5-Amino-4-cyano-3-(4-phenoxyphenyl)-1H-pyrazol-1-yl)-1-benzylpiperidine-4-carboxylic acid ethyl ester
氮氣保護下，1-苄基-4-肼基-4'-甲酸乙酯基哌啶鹽酸鹽(580mg,1.57mmol)，2-甲氧基(4-苯氧基苯基)亞甲基)丙二腈(433mg,1.57mmol)和TEA(475mg,4.71mmol)在CHCl3(20mL)中的混合物加熱回流16小時。混合物濃縮，矽膠層析柱純化，含50% EA的PE洗脫，得到黃色固體的目標產品(280mg 34.2%)。MS(ESI,m/e)[M+1]+ 521.9。 1-Benzyl-4-indolyl-4'-ethyl ester piperidine hydrochloride (580 mg, 1.57 mmol), 2-methoxy(4-phenoxyphenyl)methylene under N2 ) malononitrile (433mg, 1.57mmol) and TEA (475mg, 4.71mmol) was heated at reflux in a mixture of 3 (20mL) CHCl 16 hours. The mixture was concentrated and purified on a silica gel column eluting with 50% EtOAc eluted to afford the title product (280 mg, 34.2%). MS (ESI, m/e) [M+1] + 521.9.
步驟4：5-氨基-1-(1-苄基-4-(羥甲基)哌啶-4-基)-3-(4-苯氧基苯基)-1H-4-腈基吡唑 Step 4: 5-Amino-1-(1-benzyl-4-(hydroxymethyl)piperidin-4-yl)-3-(4-phenoxyphenyl)-1H-4-carbonitrile pyrazole
向4-(5-氨基-4-氰基-3-(4-苯氧基苯基)-1H-吡唑-1-基)-1-苄基哌啶-4-羧酸乙酯(52mg,0.1mmol)的MeOH(5mL)溶液中，加入NaBH4(8mg,0.2mmol)。10min後，加水(5mL)淬滅反應，EA(10mL×3)萃取。合併有機相，Na2SO4乾燥，濃縮，得到類白色固體產品(34mg,70.9%)。MS(ESI,m/e)[M+1]+ 480.0。 To 4-(5-amino-4-cyano-3-(4-phenoxyphenyl)-1H-pyrazol-1-yl)-1-benzylpiperidine-4-carboxylic acid ethyl ester (52 mg , 0.1 mmol) in MeOH (5mL) was added NaBH 4 (8mg, 0.2mmol). After 10 min, the reaction was quenched with water (5 mL) and EtOAc (EtOAc) The organic phases were combined, Na dried 2 SO 4, and concentrated to give an off-white solid product (34mg, 70.9%). MS (ESI, m/e) [M+1] + 480.0.
步驟5：(4-(5-氨基-4-氰基-3-(4-苯氧基苯基)-1H-吡唑)-1-基)-1-苄基哌啶-4-基)甲基甲磺酸酯 Step 5: (4-(5-Amino-4-cyano-3-(4-phenoxyphenyl)-1H-pyrazole)-1-yl)-1-benzylpiperidin-4-yl) Methyl methanesulfonate
在0℃，向5-氨基-1-(1-苄基-4-(羥甲基)哌啶-4-基)-3-(4-苯氧基苯基)-1H-4-腈基吡唑(50mg,0.1mmol)和TEA(20mg,0.20mmol)的DCM(5mL)溶液中加入MsCl(14mg,0.12mmol)。5min後，加水(5mL)淬滅反應，DCM(5mL×3)萃取。合併有機相，Na2SO4乾燥，濃縮，製備矽膠板(含10% MeOH的DCM)純化，得到黃色固體產品(35mg,62.8%)。1H NMR(400MHz,DMSO-d6)δ 7.80(d,J=8.8Hz,2H),7.45-7.36(m,2H),7.37-7.22(m,5H),7.17(t,J=7.6Hz,1H),7.11 (d,J=8.8Hz,2H),7.06(d,J=8.0Hz,2H),6.49(br s,2H),4.48(s,2H),3.40(s,2H),3.10(s,3H),2.88-2.55(m,4H),2.19-2.16(m,2H),1.92-1.86(m,2H).MS(ESI,m/e)[M+1]+ 557.9。 To 5-amino-1-(1-benzyl-4-(hydroxymethyl)piperidin-4-yl)-3-(4-phenoxyphenyl)-1H-4-carbonitrile at 0 ° C To a solution of pyrazole (50 mg, 0.1 mmol) and EtOAc (EtOAc) After 5 min, the reaction was quenched with water (5 mL)EtOAc. The organic phases were combined, Na dried 2 SO 4, concentrated and purified (DCM containing 10% MeOH in) preparing silicone plate, to give the product as a yellow solid (35mg, 62.8%). 1 H NMR (400MHz, DMSO- d 6) δ 7.80 (d, J = 8.8Hz, 2H), 7.45-7.36 (m, 2H), 7.37-7.22 (m, 5H), 7.17 (t, J = 7.6Hz , 1H), 7.11 (d, J = 8.8 Hz, 2H), 7.06 (d, J = 8.0 Hz, 2H), 6.49 (br s, 2H), 4.48 (s, 2H), 3.40 (s, 2H), 3.10 (s, 3H), 2.88-2.55 (m, 4H), 2.19-2.16 (m, 2H), 1.92-1.86 (m, 2H). MS (ESI, m/e) [M+1] + 557.9.
步驟6：1'-苄基-6-(4-苯氧基苯基)-1,2-二氫螺[咪唑並[1,2-b]吡唑-3,4'-哌啶]-7-甲腈 Step 6: 1'-Benzyl-6-(4-phenoxyphenyl)-1,2-dihydrospiro[imidazo[1,2-b]pyrazole-3,4'-piperidine]- 7-carbonitrile
(4-(5-氨基-4-氰基-3-(4-苯氧基苯基)-1H-吡唑)-1-基)-1-苄基哌啶-4-基)甲基甲磺酸酯(35mg,0.06mmol)和Cs2CO3(31mg,0.09mmol)在DMF(2mL)中的混合物加熱到50℃，反應16小時。加水(5mL)淬滅反應，EA(5mL×3)萃取。合併有機相，Na2SO4乾燥，濃縮，製備矽膠板(含10% MeOH的DCM)純化，得到黃色固體產品(12mg,43.2%)。1H NMR(400MHz,DMSO-d6)δ 7.79(d,J=8.4Hz,2H),7.43-7.40(m,3H),7.38-7.29(m,4H),7.26(br s,1H),7.19(t,J=7.6Hz,1H),7.12-7.05(m,4H),3.85(s,2H),3.53(s,2H),2.89-2.80(m,2H),2.22-2.12(m,2H),2.10-1.96(m,2H),1.85-1.75(m,2H).MS(ESI,m/e)[M+1]+ 462.0。 (4-(5-Amino-4-cyano-3-(4-phenoxyphenyl)-1H-pyrazole)-1-yl)-1-benzylpiperidin-4-yl)methylmethyl A mixture of the sulfonate (35 mg, 0.06 mmol) and Cs 2 CO 3 (31 mg, 0.09 mmol) in DMF (2 mL) was warmed to 50 < The reaction was quenched with water (5 mL) and EtOAc (EtOAc) The organic phases were combined, Na dried 2 SO 4, concentrated and purified (DCM containing 10% MeOH in) preparing silicone plate, to give the product as a yellow solid (12mg, 43.2%). 1 H NMR (400MHz, DMSO- d 6) δ 7.79 (d, J = 8.4Hz, 2H), 7.43-7.40 (m, 3H), 7.38-7.29 (m, 4H), 7.26 (br s, 1H), 7.19 (t, J = 7.6 Hz, 1H), 7.12-7.05 (m, 4H), 3.85 (s, 2H), 3.53 (s, 2H), 2.89-2.80 (m, 2H), 2.22. 2H), 2.10 - 1.96 (m, 2H), </ RTI></RTI></RTI></RTI><RTIgt;
步驟7：1'-苄基-6-(4-苯氧基苯基)-1,2-二氫螺[咪唑並[1,2-b]吡唑-3,4'-哌啶]-7-甲醯胺 Step 7: 1'-Benzyl-6-(4-phenoxyphenyl)-1,2-dihydrospiro[imidazo[1,2-b]pyrazole-3,4'-piperidine]- 7-methylamine
目標產品是由1'-苄基-6-(4-苯氧基苯基)-1,2-二氫螺[咪唑並[1,2-b]吡唑-3,4'-哌啶]-7-甲腈，根據化合物2的步驟2的類似方法合成。1H NMR(400MHz,DMSO-d6)δ 7.63(d,J=8.4Hz,2H),7.45-7.39(m,3H),7.37-7.31(m,4H),7.16(t,J=7.6Hz,1H),7.12-6.94(m,4H),6.43(s,1H),3.78(s,2H),3.55(br s,2H),2.89-2.82(m,2H),2.15-2.11(m,2H),1.84-1.72(m,2H).MS(ESI,m/e)[M+1]+ 479.9。 The target product is 1'-benzyl-6-(4-phenoxyphenyl)-1,2-dihydrospiro[imidazo[1,2-b]pyrazole-3,4'-piperidine] -7-carbonitrile was synthesized according to a similar procedure of Step 2 of Compound 2 . 1 H NMR (400MHz, DMSO- d 6) δ 7.63 (d, J = 8.4Hz, 2H), 7.45-7.39 (m, 3H), 7.37-7.31 (m, 4H), 7.16 (t, J = 7.6Hz , 1H), 7.12-6.94 (m, 4H), 6.43 (s, 1H), 3.78 (s, 2H), 3.55 (br s, 2H), 2.89-2.82 (m, 2H), 2.15-2.11 (m, 2H), 1.84-1.72 (m, 2H). MS (ESI, m/e) [M+1] + 479.9.
化合物39：6-(4-苯氧基苯基)-1,2-二氫螺[咪唑並[1,2-b]吡唑-3,4'-哌啶]-7-甲醯胺 Compound 39 : 6-(4-phenoxyphenyl)-1,2-dihydrospiro[imidazo[1,2-b]pyrazole-3,4'-piperidine]-7-carboxamide
向1'-苄基-6-(4-苯氧基苯基)-1,2-二氫螺[咪唑並[1,2-b]吡唑-3,4'-哌啶]-7-甲醯胺(50mg,0.10mmol)的MeOH(10mL)溶液中，加入10% w/w Pd(OH)2/C(5mg)和HOAc(1d)。混合物在氫氣下攪拌16小時，過濾，濾液濃縮，得到黃色固體產品(20mg,51.4%)。MS(ESI,m/e)[M+1]+ 390.0。 To 1'-benzyl-6-(4-phenoxyphenyl)-1,2-dihydrospiro[imidazo[1,2-b]pyrazole-3,4'-piperidine]-7- To a solution of methotrexate (50 mg, 0.10 mmol) in MeOH (10 mL), 10% w/w Pd(OH) 2 / C (5 mg) and HOAc (1d). The mixture was stirred with mp EtOAc (EtOAc)EtOAc. MS (ESI, m/e) [M+1] + 39.
化合物40：1'-丙烯醯基-6-(4-苯氧基苯基)-1,2-二氫螺[咪唑並[1,2-b]吡唑-3,4'-哌啶]-7-甲醯胺 Compound 40 : 1'-Prodecyl-6-(4-phenoxyphenyl)-1,2-dihydrospiro[imidazo[1,2-b]pyrazole-3,4'-piperidine] -7-formamide
化合物40是由化合物39和丙烯醯氯根據，化合物8的類似方法合成。1H NMR(400MHz,DMSO-d6)δ 7.64(d,J=8.8Hz,2H),7.44-7.37(m,2H),7.16(t,J=7.6Hz,1H),7.05(d,J=7.6Hz,2H),6.97(d,J=8.8Hz,2H),6.86(dd,J=10.5,16.7Hz,1H),6.51(br s,1H),6.12(dd,J=2.3,16.7Hz,1H),5.69(dd,J=2.3,10.5Hz,1H),4.13-3.95(m,2H),3.83(s,2H),3.60-3.38(m,2H),1.99-1.76(m,4H).MS(ESI,m/e)[M+1]+ 443.9。 The compound 40 is similar to the synthesis of compound 8 from compound 39 and Bing Xixi chlorine. 1 H NMR (400 MHz, DMSO-d 6 ) δ 7.64 (d, J = 8.8 Hz, 2H), 7.44 - 7.37 (m, 2H), 7.16 (t, J = 7.6 Hz, 1H), 7.05 (d, J = 7.6 Hz, 2H), 6.97 (d, J = 8.8 Hz, 2H), 6.86 (dd, J = 10.5, 16.7 Hz, 1H), 6.51 (br s, 1H), 6.12 (dd, J = 2.3, 16.7 Hz, 1H), 5.69 (dd, J = 2.3, 10.5 Hz, 1H), 4.13 - 3.95 (m, 2H), 3.83 (s, 2H), 3.60-3.38 (m, 2H), 1.99-1.76 (m, 4H). MS (ESI, m/e) [M+1] + 443.9.
實施例18：化合物41到43的合成Example 18: Synthesis of Compounds 41 to 43
化合物41：1-苄基-2'-(4-苯氧基苯基)-5',6'-二氫-4'H-螺[哌啶-4,7'-吡唑並[1,5-a]嘧啶]-3'-甲醯胺 Compound 41 : 1-benzyl-2'-(4-phenoxyphenyl)-5',6'-dihydro-4'H-spiro[piperidine-4,7'-pyrazolo[1, 5-a]pyrimidin]-3'-formamide
步驟1：2-(1-苄基哌啶-4-亞基)乙酸乙酯 Step 1: 2-(1-Benzylpiperidin-4-ylidene)acetate
在0℃下，向NaH(318mg,7.94mmol)的THF(20mL)懸浮液中滴加磷醯基乙酸三乙酯(1.78g,7.94mmol)的THF(5mL)溶液，30min內滴畢。攪拌10min後，再滴加1-苄 基哌啶-4-酮(1.0g,5.29mmol)的THF(5mL)溶液，20min內滴畢。混合物攪拌60min，加水(10mL)淬滅。EA(10mL×3)萃取，合併有機相，Na2SO4乾燥，濃縮，矽膠層析柱純化，含25% EA的PE洗脫，得到黃色油狀產品(1.2g,87.3%)。MS(ESI,m/e)[M+1]+ 260.0。 A solution of triethyl phosphonium acetate (1.78 g, 7.94 mmol) in THF (5 mL) was added dropwise EtOAc. After stirring for 10 min, a solution of 1-benzylpiperidin-4-one (1.0 g, 5.29 mmol) in THF (5 mL). The mixture was stirred for 60 min and quenched with water (10 mL). EA (10mL × 3). The combined organic phases were dried Na 2 SO 4, concentrated and purified by column chromatography on silica gel, containing 25% EA in PE to give the product as a yellow oil (1.2g, 87.3%). MS (ESI, m/e) [M+1] + 260.0.
步驟2：1-苄基-5'-氧代-2'-(4-苯氧基苯基)-5',6'-二氫-4'H-螺[哌啶-4,7'-吡唑並[1,5-a]嘧啶]-3'-甲腈 Step 2: 1-Benzyl-5'-oxo-2'-(4-phenoxyphenyl)-5',6'-dihydro-4'H-spiro[piperidine-4,7'- Pyrazolo[1,5-a]pyrimidin]-3'-carbonitrile
氮氣保護下，5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈(1.0g，3.6mmol)，2-(1-苄基哌啶-4-亞基)乙酸乙酯(1.1g,4.3mmol)和K2CO3(745mg,5.4mmol)在DMF(20mL)中的混合物，加熱至80℃，攪拌16小時。然後，加水(20mL)淬滅反應，EA(20mL×3)萃取，合併有機相，Na2SO4乾燥，濃縮，矽膠層析柱純化，含30% EA的PE洗脫，得到黃色固體產品(950mg,54.9%)。1H NMR(400MHz,DMSO-d6)δ 11.92(s,1H),7.84(d,J=8.8Hz,2H),7.48-7.40(m,2H),7.36-7.29(m,4H),7.27-7.23(m,1H),7.20(t,J=7.6Hz,1H),7.15-7.07(m,4H),3.55(s,2H),3.01(s,2H),2.81-2.73(m,2H),2.39-2.27(m,2H),2.26-2.16(m,2H),1.83-1.74(m,2H).MS(ESI,m/e)[M+1]+ 489.9。 5-Amino-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitrile (1.0 g, 3.6 mmol), 2-(1-benzylpiperidin-4-y. yl) acetate (1.1g, 4.3mmol) and K 2 CO 3 (745mg, 5.4mmol ) in the mixture (20mL) DMF, was heated to 80 ℃, stirred for 16 hours. Then, water (20mL) quenched the reaction, EA (20mL × 3). The combined organic phases were dried Na 2 SO 4, concentrated and purified by column chromatography on silica gel, containing 30% EA in PE to give the product as a yellow solid ( 950mg, 54.9%). 1 H NMR (400MHz, DMSO- d 6) δ 11.92 (s, 1H), 7.84 (d, J = 8.8Hz, 2H), 7.48-7.40 (m, 2H), 7.36-7.29 (m, 4H), 7.27 -7.23 (m, 1H), 7.20 (t, J = 7.6 Hz, 1H), 7.15-7.07 (m, 4H), 3.55 (s, 2H), 3.01 (s, 2H), 2.81-2.73 (m, 2H) ), 2.39-2.27 (m, 2H), 2.26-2.16 (m, 2H), 1.83-1.74 (m, 2H). MS (ESI, m/e) [M+1] + 489.9.
步驟3：1-苄基-5'-氧代-2'-(4-苯氧基苯基)-5',6'-二氫-4'H-螺[哌啶-4,7'-吡唑並[1,5-a]嘧啶]-3'-甲醯胺 Step 3: 1-Benzyl-5'-oxo-2'-(4-phenoxyphenyl)-5',6'-dihydro-4'H-spiro[piperidine-4,7'- Pyrazolo[1,5-a]pyrimidin]-3'-formamide
1-苄基-5'-氧代-2'-(4-苯氧基苯基)-5',6'-二氫-4'H-螺[哌啶-4,7'-吡唑並[1,5-a]嘧啶]-3'-甲腈(500mg,1.02mmol)的H3PO4(5mL)溶液，加熱至130度，反應1小時。將混合物倒入水(20mL)中，EA(20mL×3)萃取，合併有機相，Na2SO4乾燥，濃縮，矽膠層析柱純化，含5% MeOH的DCM洗脫，得到黃色固體產品(180mg,34.8%)。MS(ESI,m/e)[M+1]+ 507.9。 1-benzyl-5'-oxo-2'-(4-phenoxyphenyl)-5',6'-dihydro-4'H-spiro[piperidine-4,7'-pyrazole [1,5-a] pyrimidine] -3'-carbonitrile (500mg, 1.02mmol) in H 3 PO 4 (5mL) was heated to 130 degrees for 1 hour. The mixture was poured into water (20mL) in, EA (20mL × 3). The combined organic phases were dried 2 SO 4 Na, concentrated and purified by column chromatography on silica gel, containing 5% MeOH in DCM to give the product as a yellow solid ( 180 mg, 34.8%). MS (ESI, m/e) [M+1] + 507.9.
步驟3：1-苄基-2'-(4-苯氧基苯基)-5',6'-二氫-4'H-螺[哌啶-4,7'-吡唑並[1,5-a]嘧啶]-3'-甲醯胺 Step 3: 1-Benzyl-2'-(4-phenoxyphenyl)-5',6'-dihydro-4'H-spiro[piperidine-4,7'-pyrazolo[1, 5-a]pyrimidin]-3'-formamide
1-苄基-5'-氧代-2'-(4-苯氧基苯基)-5',6'-二氫-4'H-螺[哌啶-4,7'-吡唑並[1,5-a]嘧啶]-3'-甲醯胺(180mg,0.36mmol)的BH3/THF(1N,20mL)溶液加熱回流3小時。加入MeOH(20mL)和濃鹽酸(2mL)淬滅反應。將混合物在60℃攪拌1小時，NaHCO3鹼化，EA(20mL×3)萃取，合併有機相，Na2SO4乾燥，濃縮，矽膠層析柱純化，含5% MeOH的DCM洗脫，得到黃 色固體產品(120mg,67.6%)。1H NMR(400MHz,CD3OD-d4)δ 7.39-7.22(m,9H),7.09-7.04(m,1H),6.99-6.94(m,4H),3.86(s,2H),3.35(t,J=5.6Hz,2H),3.19-3.11(m,2H),2.80-2.66(m,2H),2.50-2.40(m,2H),2.10(t,J=5.6Hz,2H),1.87-1.78(m,2H).MS(ESI,m/e)[M+1]+ 493.9。 1-benzyl-5'-oxo-2'-(4-phenoxyphenyl)-5',6'-dihydro-4'H-spiro[piperidine-4,7'-pyrazole [1,5-a] pyrimidine] -3'-acyl amine (180mg, 0.36mmol) of BH 3 / THF (1N, 20mL ) was refluxed for 3 hours. The reaction was quenched by the addition of MeOH (20 mL)EtOAc. The mixture was stirred at 60 ℃ 1 hours, NaHCO 3 was basified, EA (20mL × 3). The combined organic phases were dried Na 2 SO 4, concentrated and purified by column chromatography on silica gel, containing 5% MeOH in DCM to afford Yellow solid product (120 mg, 67.6%). 1 H NMR (400MHz, CD 3 OD-d 4) δ 7.39-7.22 (m, 9H), 7.09-7.04 (m, 1H), 6.99-6.94 (m, 4H), 3.86 (s, 2H), 3.35 ( t, J = 5.6Hz, 2H) , 3.19-3.11 (m, 2H), 2.80-2.66 (m, 2H), 2.50-2.40 (m, 2H), 2.10 (t, J = 5.6Hz, 2H), 1.87 -1.78 (m, 2H). MS (ESI, m/e) [M+1] + 493.9.
化合物42：2'-(4-苯氧基苯基)-5',6'-二氫-4'H-螺[哌啶-4,7'-吡唑並[1,5-a]嘧啶]-3'-甲醯胺三氟乙酸鹽 Compound 42 : 2'-(4-phenoxyphenyl)-5',6'-dihydro-4'H-spiro[piperidine-4,7'-pyrazolo[1,5-a]pyrimidine ]-3'-carbamidine trifluoroacetate
化合物42是由1-苄基-2'-(4-苯氧基苯基)-5',6'-二氫-4'H-螺[哌啶-4,7'-吡唑並[1,5-a]嘧啶]-3'-甲醯胺，根據化合物39的類似方法合成。1H NMR(400MHz,DMSO-d6)δ 8.66(br s,2H),7.51(d,J=8.6Hz,2H),7.46-7.37(m,2H),7.18(t,J=7.6Hz,1H),7.11-7.04(m,4H),6.79(s,1H),3.40-3.33(m,4H),3.17-3.06(m,2H),2.46-2.35(m,2H),2.17-2.10(m,2H),1.96-1.87(m,2H).MS(ESI,m/e)[M+1]+ 403.9。 Compound 42 from 1-benzyl-2 '- (4-phenoxyphenyl) -5', 6'-dihydro-spiro -4'H- [piperidine4,7' pyrazolo [1 , 5-a]pyrimidin]-3'-formamide, synthesized according to a similar method to compound 39 . 1 H NMR (400MHz, DMSO- d 6) δ 8.66 (br s, 2H), 7.51 (d, J = 8.6Hz, 2H), 7.46-7.37 (m, 2H), 7.18 (t, J = 7.6Hz, 1H), 7.11-7.04 (m, 4H), 6.79 (s, 1H), 3.40-3.33 (m, 4H), 3.17-3.06 (m, 2H), 2.46-2.35 (m, 2H), 2.17-2.10 ( m, 2H), 1.96-1.87 (m, 2H). MS (ESI, m/e) [M+1] + 403.9.
化合物43：1-丙烯醯基-2'-(4-苯氧基苯基)-5',6'-二氫-4'H-螺[哌啶-4,7'-吡唑並[1,5-a]嘧啶]-3'-甲醯胺 Compound 43 : 1-propenylfluorenyl-2'-(4-phenoxyphenyl)-5',6'-dihydro-4'H-spiro[piperidine-4,7'-pyrazolo[1] ,5-a]pyrimidine]-3'-formamide
化合物43是由化合物42和丙烯醯氯，根據化合物8的類似方法合成。1HNMR(400MHz,DMSO-d6)δ 7.50(d,J=8.8Hz,2H),7.45-7.37(m,2H),7.17(t,J=.2Hz,1H),7.12-7.01(m,4H),6.85(dd,J=10.4,16.7Hz,1H),6.73(s,1H),6.11(dd,J=2.4,16.7Hz,1H),5.68(dd,J=2.4,10.4Hz,1H),4.23(d,J=13.2Hz,1H),4.03(d,J=13.2Hz,1H),3.43(t,J=12.0Hz,1H),3.37-3.33(m,2H),3.16(t,J=12.0,1H),2.24-2.08(m,4H),1.82-1.73(m,2H).MS(ESI,m/e)[M+1]+ 457.9。 Compound 43 was synthesized from Compound 42 and propylene chloride in a similar manner to Compound 8 . 1 H NMR (400 MHz, DMSO-d 6 ) δ 7.50 (d, J = 8.8 Hz, 2H), 7.45-7.37 (m, 2H), 7.17 (t, J = .2 Hz, 1H), 7.12-7.01 (m, 4H), 6.85 (dd, J = 10.4, 16.7 Hz, 1H), 6.73 (s, 1H), 6.11 (dd, J = 2.4, 16.7 Hz, 1H), 5.68 (dd, J = 2.4, 10.4 Hz, 1H) ), 4.23 (d, J = 13.2 Hz, 1H), 4.03 (d, J = 13.2 Hz, 1H), 3.43 (t, J = 12.0 Hz, 1H), 3.37 - 3.33 (m, 2H), 3.16 (t) , J =12.0, 1H), 2.24-2.08 (m, 4H), 1.82-1.73 (m, 2H). MS (ESI, m/e) [M+1] + 457.9.
實施例19：化合物44到46的合成Example 19: Synthesis of Compounds 44 to 46
化合物44：1-苄基-6'-(4-苯氧基苯基)-1',2'-二氫螺[氮雜環丁烷-3,3'-咪唑並[1,2-b]吡唑]-7'-甲醯胺 Compound 44 : 1-benzyl-6'-(4-phenoxyphenyl)-1',2'-dihydrospiro[azetidin-3,3'-imidazo[1,2-b Pyrazole]-7'-formamide
步驟1：1-苄基-3-羧酸甲酯氮雜環丁烷 Step 1: 1-Benzyl-3-carboxylic acid methyl ester azetidine
在0℃下，向氮雜環丁烷-3-羧酸甲酯(5.0g,33.1mmol)和DIEA(10.7g,82.8mmol)的DMF(50mL)溶液中滴加溴苄(5.7g,33.1mmol)，約10min滴完。室溫攪拌2小時，將混合物倒入水(50mL)中，EA(50mL×3)萃取。合併有機相，Na2SO4乾燥，濃縮，矽膠層析柱純化(EA/PE=1/4)，得到 黃色油狀產品(3.5g,51.6%)。1H NMR(DMSO-d6)δ 7.34-7.20(m,5H),3.62(s,3H),3.53(s,2H),3.41-3.35(m,1H),3.29-3.31(m,2H),3.19-3.22(m,2H).MS(ESI,m/e)[M+1]+ 206.0。 To a solution of methyl azetidine-3-carboxylate (5.0 g, 33.1 mmol) and DIEA (10.7 g, 82.8 mmol) in DMF (50 mL) Mmmol), about 10 minutes after the completion of the drop. After stirring at room temperature for 2 hours, the mixture was poured into water (50 mL) The organic phases were combined, dried Na 2 SO 4, concentrated and purified by column chromatography on silica gel (EA / PE = 1/4 ), to give the product as a yellow oil (3.5g, 51.6%). 1 H NMR (DMSO-d 6 ) δ 7.34-7.20 (m, 5H), 3.62 (s, 3H), 3.53 (s, 2H), 3.41-3.35 (m, 1H), 3.29-3.31 (m, 2H) , 3.19-3.22 (m, 2H). MS (ESI, m/e) [M+1] + 206.0.
步驟2：1-(1-苄基-3-(甲氧基羰基)氮雜環丁烷-3-基)肼-1,2-二羧酸二叔丁酯 Step 2: 1-(1-Benzyl-3-(methoxycarbonyl)azetidin-3-yl)indole-1,2-dicarboxylic acid di-tert-butyl ester
目標產物由1-苄基-3-羧酸甲酯氮雜環丁烷和偶氮二甲酸二叔丁酯，根據化合物38的步驟1的類似方法合成。MS(ESI,m/e)[M+1]+ 435.9。 The title product was synthesized from a similar procedure of Step 1 of Compound 38 from 1-benzyl-3-carboxylic acid methyl ester azetidine and di-tert-butyl azodicarboxylate. MS (ESI, m/e) [M+1] + 435.9.
步驟3到5：5-氨基-1-(1-苄基-3-(羥甲基)氮雜環丁烷-3-基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈 Steps 3 to 5: 5-amino-1-(1-benzyl-3-(hydroxymethyl)azetidin-3-yl)-3-(4-phenoxyphenyl)-1H-pyridyl Oxazole-4-carbonitrile
目標產物以1-(1-苄基-3-(甲氧基羰基)氮雜環丁烷-3-基)肼-1,2-二羧酸二叔丁酯為起始原料，根據化合物38的步驟2到4的類似方法合成。1H NMR(DMSO-d6)δ 7.78(d,J=8.8Hz,2H),7.46-7.38(m,2H),7.36-7.21(m,5H),7.18(t,J=7.6Hz,1H),7.11-7.04(m,4H),6.17(s,2H),5.50(t,J=5.2Hz,1H), 3.89(d,J=5.2Hz,2H),3.61-3.63(m,4H),3.38(d,J=6.4Hz,2H).MS(ESI,m/e)[M+1]+ 451.9。 The target product is starting from 1-(1-benzyl-3-(methoxycarbonyl)azetidin-3-yl)indole-1,2-dicarboxylic acid di-tert-butyl ester, according to compound 38 A similar method of steps 2 to 4 is synthesized. 1 H NMR (DMSO-d 6 ) δ 7.78 (d, J = 8.8 Hz, 2H), 7.46-7.38 (m, 2H), 7.36-7.21 (m, 5H), 7.18 (t, J = 7.6 Hz, 1H) ), 7.11-7.04 (m, 4H), 6.17 (s, 2H), 5.50 (t, J = 5.2 Hz, 1H), 3.89 (d, J = 5.2 Hz, 2H), 3.61-3.63 (m, 4H) , 3.38 (d, J = 6.4 Hz, 2H). MS (ESI, m/e) [M+1] + 451.9.
步驟6：(3-(5-氨基-4-氰基-3-(4-苯氧基苯基)-1H-吡唑-1-基)-1-苄基氮雜環丁烷-3-基)甲基甲磺酸酯 Step 6: (3-(5-Amino-4-cyano-3-(4-phenoxyphenyl)-1H-pyrazol-1-yl)-1-benzylazetidin-3- Methyl methanesulfonate
目標產物由5-氨基-1-(1-苄基-3-(羥甲基)氮雜環丁烷-3-基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈，根據化合物38的步驟5的類似方法合成。1H NMR(DMSO-d6)δ 7.82-7.74(m,2H),7.46-7.38(m,2H),7.36-7.23(m,5H),7.21-7.15(m,1H),7.13-7.04(m,4H),6.52(s,2H),4.69(s,2H),3.70(d,J=8.4Hz,2H),3.66(s,2H),3.49(d,J=8.4Hz,2H),3.13(s,3H).MS(ESI,m/e)[M+1]+ 529.9。 The target product consists of 5-amino-1-(1-benzyl-3-(hydroxymethyl)azetidin-3-yl)-3-(4-phenoxyphenyl)-1H-pyrazole- 4-carbonitrile was synthesized according to a similar procedure as in Step 5 of Compound 38 . 1 H NMR (DMSO-d 6 ) δ 7.82-7.74 (m, 2H), 7.46-7.38 (m, 2H), 7.36-7.23 (m, 5H), 7.21-7.15 (m, 1H), 7.13-7.04 ( m, 4H), 6.52 (s, 2H), 4.69 (s, 2H), 3.70 (d, J = 8.4 Hz, 2H), 3.66 (s, 2H), 3.49 (d, J = 8.4 Hz, 2H), 3.13 (s, 3H). MS (ESI, m/e) [M+1] + 529.9.
步驟7,8：1-苄基-6'-(4-苯氧基苯基)-1',2'-二氫螺[氮雜環丁烷-3,3'-咪唑並[1,2-b]吡唑]-7'-甲醯胺 Step 7, 8: 1-Benzyl-6'-(4-phenoxyphenyl)-1',2'-dihydrospiro[azetidin-3,3'-imidazo[1,2 -b]pyrazole]-7'-formamide
目標產物由(3-(5-氨基-4-氰基-3-(4-苯氧基苯基)-1H-吡唑-1-基)-1-苄基氮雜環丁烷-3-基)甲基甲磺酸酯，根據化合物38的 步驟6到7的類似方法合成。1H NMR(DMSO-d6)δ 7.66(d,J=8.8Hz,2H),7.46-7.38(m,2H),7.35-7.28(m,4H),7.28-7.21(m,1H),7.16(t,J=7.6Hz,1H),7.06(d,J=8.0Hz,2H),7.01(d,J=8.8Hz,2H),6.52(s,1H),4.19(s,2H),3.67(s,2H),3.54(s,4H).MS(ESI,m/e)[M+1]+ 451.9。 The target product consists of (3-(5-amino-4-cyano-3-(4-phenoxyphenyl)-1H-pyrazol-1-yl)-1-benzylazetidin-3- Methyl methanesulfonate was synthesized according to a similar procedure to Steps 6 to 7 of Compound 38 . 1 H NMR (DMSO-d 6 ) δ 7.66 (d, J = 8.8 Hz, 2H), 7.46-7.38 (m, 2H), 7.35-7.28 (m, 4H), 7.28-7.21 (m, 1H), 7.16 (t, J = 7.6 Hz, 1H), 7.06 (d, J = 8.0 Hz, 2H), 7.01 (d, J = 8.8 Hz, 2H), 6.52 (s, 1H), 4.19 (s, 2H), 3.67 (s, 2H), 3.54 (s, 4H). MS (ESI, m/e) [M+1] + 451.9.
化合物45：6'-(4-苯氧基苯基)-1',2'-二氫螺[氮雜環丁烷-3,3'-咪唑並[1,2-b]吡唑]-7'-甲醯胺 Compound 45: 6'-(4-phenoxyphenyl)-1',2'-dihydrospiro[azetidin-3,3'-imidazo[1,2-b]pyrazole]- 7'-carbamamine
目標產物由化合物44，根據化合物39的類似方法合成。1H NMR(DMSO-d6)δ 7.67(d,J=8.4Hz,2H),7.46-7.36(m,2H),7.16(t,J=7.6Hz,1H),7.06(d,J=7.6Hz,2H),7.00(d,J=8.4Hz,2H),6.62(s,1H),4.19-4.20(m,4H),3.78(d,J=9.6Hz,2H).MS(ESI,m/e)[M+1]+ 361.9。 The title product was synthesized from compound 44 in a similar manner to compound 39 . 1 H NMR (DMSO-d 6 ) δ 7.67 (d, J = 8.4 Hz, 2H), 7.46-7.36 (m, 2H), 7.16 (t, J = 7.6 Hz, 1H), 7.06 (d, J = 7.6 Hz, 2H), 7.00 (d, J = 8.4 Hz, 2H), 6.62 (s, 1H), 4.19-4.20 (m, 4H), 3.78 (d, J = 9.6 Hz, 2H). MS (ESI, m /e)[M+1] + 361.9.
化合物46：1-丙烯醯基-6'-(4-苯氧基苯基)-1',2'-二氫螺[氮雜環丁烷-3,3'-咪唑並[1,2-b]吡唑]-7'-甲醯胺 Compound 46: 1-propenylfluorenyl-6'-(4-phenoxyphenyl)-1',2'-dihydrospiro[azetidin-3,3'-imidazo[1,2- b]pyrazole]-7'-formamide
目標產物由化合物45和丙烯醯氯，根據化合物8的類似方法合成。1H NMR(DMSO-d6)δ 7.66(d,J=8.8Hz,2H), 7.45-7.37(m,2H),7.16(t,J=7.6Hz,1H),7.06(d,J=7.6Hz,2H),7.00(d,J=8.8Hz,2H),6.62(s,1H),6.36(dd,J=17.0,10.3Hz,1H),6.15(dd,J=17.0,2.1Hz,1H),5.72(dd,J=10.3,2.1Hz,1H),4.62(d,J=9.6Hz,1H),4.56(d,J=9.6Hz,1H),4.32(d,J=11.2Hz,1H),4.27(d,J=11.2Hz,1H),4.22(s,2H).MS(ESI,m/e)[M+1]+ 415.9。 The target product was synthesized from Compound 45 and propylene sulfonium chloride in a similar manner to Compound 8 . 1 H NMR (DMSO-d 6 ) δ 7.66 (d, J = 8.8 Hz, 2H), 7.45-7.37 (m, 2H), 7.16 (t, J = 7.6 Hz, 1H), 7.06 (d, J = 7.6 Hz, 2H), 7.00 (d, J = 8.8 Hz, 2H), 6.62 (s, 1H), 6.36 (dd, J = 17.0, 10.3 Hz, 1H), 6.15 (dd, J = 17.0, 2.1 Hz, 1H) ), 5.72 (dd, J = 10.3, 2.1 Hz, 1H), 4.62 (d, J = 9.6 Hz, 1H), 4.56 (d, J = 9.6 Hz, 1H), 4.32 (d, J = 11.2 Hz, 1H) ), 4.27 (d, J = 11.2 Hz, 1H), 4.22 (s, 2H). MS (ESI, m/e) [M+1] + 415.9.
實施例20：化合物47-50的合成Example 20: Synthesis of Compound 47-50
化合物47：2-(3-氨基苯基)-6-(4-苯氧基苯基)-1H-咪唑並[1,2-b]吡唑-7-甲醯胺 Compound 47 : 2-(3-Aminophenyl)-6-(4-phenoxyphenyl)-1H-imidazo[1,2-b]pyrazole-7-carboxamide
步驟1：5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺 Step 1: 5-Amino-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide
5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈(1.0g,3.6mmol)的H3PO4(20ml)溶液，加熱至120℃反應4小時。將混合物倒入水(100mL)中，EA(100mL×3)萃取。合併有機層，Na2SO4乾燥，濃縮得到(850mg,77.5%)黃色固體產物。MS(ESI,m/e)[M+1]+ 295.1。 5-amino-3- (4-phenoxyphenyl) lH-pyrazole-4-carbonitrile (1.0g, 3.6mmol) in H 3 PO 4 (20ml) was heated to 120 deg.] C for 4 hours. The mixture was poured into water (100 mL) and EA (100 mL×3). The organic layers were combined, Na dried 2 SO 4, and concentrated to give (850mg, 77.5%) as a yellow solid. MS (ESI, m/e) [M+1] + 295.1.
步驟2：2-(3-硝基苯基)-6-(4-苯氧基苯基)-1H-咪唑並[1,2-b]吡唑-7-甲醯胺 Step 2: 2-(3-Nitrophenyl)-6-(4-phenoxyphenyl)-1H-imidazo[1,2-b]pyrazole-7-carboxamide
5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺(29.4mg,0.1mmol)和2-溴-1-(3-硝基苯基)乙酮(24.4mg,0.1mmol)在EtOH(2mL)中的混合物，在80℃攪拌16小時。將混合物過濾，得到黃色固體粗品2-(3-硝基苯基)-6-(4-苯氧基苯基)-1H-咪唑並[1,2-b]吡唑-7-甲醯胺(5mg)。MS(ESI)m/e[M+1]+ 440.0。 5-amino-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide (29.4 mg, 0.1 mmol) and 2-bromo-1-(3-nitrophenyl)ethanone (24.4 mg, 0.1 mmol) in EtOAc (2 mL) EtOAc. The mixture was filtered to give a crude yellow solid, 2-(3-nitrophenyl)-6-(4-phenoxyphenyl)-1H-imidazo[1,2-b]pyrazole-7-carboxamide (5mg). MS (ESI) m / e [M+1] + 440.0.
步驟3：2-(3-氨基苯基)-6-(4-苯氧基苯基)-1H-咪唑並[1,2-b]吡唑-7-甲醯胺 Step 3: 2-(3-Aminophenyl)-6-(4-phenoxyphenyl)-1H-imidazo[1,2-b]pyrazole-7-carboxamide
向2-(3-硝基苯基)-6-(4-苯氧基苯基)-1H-咪唑並[1,2-b]吡唑-7-甲醯胺(600mg,1.37mmol)的MeOH(10mL)和DCM(10mL)溶液中，加10% w/w Pd/C(100mg)。在H2條件下，室溫攪拌4小時。將混合物過濾。濃縮濾液，製備HPLC純化，水相含0.1% TFA，30%到90% CH3CN梯度洗脫，收集合併目標產品，過夜凍幹，得到白色固體2-(3-氨基苯基)-6-(4-苯氧基苯基)-1H-咪唑並[1,2-b]吡唑-7-甲醯胺(73mg,13%)。1H NMR(400 MHz,DMSO-d6)δ 12.03(d,J=10.4Hz,1H),8.14(d,J=8.0Hz,1H),7.73(d,J=8.4Hz,2H),7.44(d,J=7.6Hz,1H),7.42(d,J=7.6Hz,1H),7.38-7.26(m,3H),7.18(t,J=7.6Hz,1H),7.10(d,J=8.0Hz,2H),7.03(d,J=8.4Hz,2H),6.98-6.86(m,2H).MS(ESI)m/e[M+1]+ 409.9。 To 2-(3-nitrophenyl)-6-(4-phenoxyphenyl)-1H-imidazo[1,2-b]pyrazole-7-carboxamide (600 mg, 1.37 mmol) To a solution of MeOH (10 mL) and DCM (10 mL), 10% w/w Pd / C (100 mg). The mixture was stirred at room temperature for 4 hours under H 2 conditions. The mixture was filtered. The filtrate was concentrated, purified by preparative HPLC, and the aqueous phase was eluted with 0.1% TFA, 30% to 90% CH 3 CN gradient eluted, and the combined product was collected and lyophilized overnight to give 2-(3-aminophenyl)-6- as a white solid. (4-Phenoxyphenyl)-1H-imidazo[1,2-b]pyrazole-7-carboxamide (73 mg, 13%). 1 H NMR (400 MHz, DMSO-d 6 ) δ 12.03 (d, J = 10.4 Hz, 1H), 8.14 (d, J = 8.0 Hz, 1H), 7.73 (d, J = 8.4 Hz, 2H), 7.44 (d, J = 7.6 Hz, 1H), 7.42 (d, J = 7.6 Hz, 1H), 7.38-7.26 (m, 3H), 7.18 (t, J = 7.6 Hz, 1H), 7.10 (d, J = 8.0 Hz, 2H), 7.03 (d, J = 8.4 Hz, 2H), 6.98-6.86 (m, 2H). MS (ESI) m/e [M+1] + 409.9.
化合物48：2-(3-丙烯醯胺基苯基)-6-(4-苯氧基苯基)-1H-咪唑並[1,2-b]吡唑-7-甲醯胺 Compound 48 : 2-(3-Acrylaminophenyl)-6-(4-phenoxyphenyl)-1H-imidazo[1,2-b]pyrazole-7-carboxamide
目標產物由2-(3-氨基苯基)-6-(4-苯氧基苯基)-1H-咪唑並[1,2-b]吡唑-7-甲醯胺和丙烯醯氯，根據化合物8的類似方法合成。1H NMR(400MHz,CD3OD-d4)δ 8.06(s,1H),7.83(s,1H),7.64(d,J=8.4Hz,1H),7.52-7.38(m,5H),7.15(t,J=7.6Hz,1H),7.09-7.05(m,4H),6.49-6.37(m,2H),5.80(dd,J=4.0,8.8Hz,1H).MS(ESI)m/e[M+1]+ 463.9。 The target product consists of 2-(3-aminophenyl)-6-(4-phenoxyphenyl)-1H-imidazo[1,2-b]pyrazole-7-carboxamide and propylene hydrazine, according to A similar method of compound 8 was synthesized. 1 H NMR (400 MHz, CD 3 OD-d 4 ) δ 8.06 (s, 1H), 7.83 (s, 1H), 7.64 (d, J = 8.4 Hz, 1H), 7.52-7.38 (m, 5H), 7.15 (t, J = 7.6Hz, 1H ), 7.09-7.05 (m, 4H), 6.49-6.37 (m, 2H), 5.80 (dd, J = 4.0,8.8Hz, 1H) .MS (ESI) m / e [M+1] + 463.9.
化合物49：3-(3-氨基苯基)-6-(4-苯氧基苯基)-1H-吡唑並[1,5-a]咪唑-7-甲醯胺 Compound 49 : 3-(3-Aminophenyl)-6-(4-phenoxyphenyl)-1H-pyrazolo[1,5-a]imidazole-7-carboxamide
目標化合物來自製備化合物48的步驟2中的另一異構體。1H NMR(400MHz,DMSO-d6)δ 12.22(d,J=2.4Hz,1H),8.03(s,1H),7.89(d,J=2.4Hz,1H),7.81-7.68(m,3H),7.46-7.37(m,4H),7.19(t,J=7.6Hz,1H),7.13-7.08(m,5H),6.98(d,J=7.6Hz,1H).MS(ESI)m/e[M+1]+ 410.1。 The target compound is from the other isomer in the second step of the preparation of compound 48 . 1 H NMR (400MHz, DMSO- d6) δ 12.22 (d, J = 2.4Hz, 1H), 8.03 (s, 1H), 7.89 (d, J = 2.4Hz, 1H), 7.81-7.68 (m, 3H) , 7.46-7.37 (m, 4H), 7.19 (t, J = 7.6 Hz, 1H), 7.13 - 7.08 (m, 5H), 6.98 (d, J = 7.6 Hz, 1H). MS (ESI) m/e [M+1] + 410.1.
化合物50：3-(3-丙烯醯胺基苯基)-6-(4-苯氧基苯基)-1H-咪唑並[1,2-b]吡唑-7-甲醯胺 Compound 50 : 3-(3-Acrylaminophenyl)-6-(4-phenoxyphenyl)-1H-imidazo[1,2-b]pyrazole-7-carboxamide
目標產物由化合物49和丙烯醯氯，根據化合物8的類似方法合成。1H NMR(400MHz,DMSO-d6)δ 12.18(d,J=2.4Hz,1H),10.31(s,1H),8.38(s,1H),7.85(d,J=8.0Hz,1H),7.80-7.73(m,4H),7.46-7.40(m,3H),7.19(t,J=8.0Hz,1H),7.13-7.07(m,4H),6.50(dd,J=10.2,17.0Hz,1H),6.27(d,J=17.0Hz,1H),5.76(d,J=10.2Hz,1H).MS(ESI)m/e[M+1]+ 463.9。 The target product was synthesized from Compound 49 and acrylofluorene chloride in a similar manner to Compound 8 . 1 H NMR (400MHz, DMSO- d 6) δ 12.18 (d, J = 2.4Hz, 1H), 10.31 (s, 1H), 8.38 (s, 1H), 7.85 (d, J = 8.0Hz, 1H), 7.80-7.73 (m, 4H), 7.46-7.40 (m, 3H), 7.19 (t, J = 8.0 Hz, 1H), 7.13 - 7.07 (m, 4H), 6.50 (dd, J = 10.2, 17.0 Hz, 1H), 6.27 (d, J = 17.0 Hz, 1H), 5.76 (d, J = 10.2 Hz, 1H). MS (ESI) m/e [M+1] + 463.9.
實施例21：化合物51到60的合成Example 21: Synthesis of Compounds 51 to 60
化合物51：2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺三氟乙酸鹽 Compound 51 : 2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[4,5-c Pyridine-3-carboxamide trifluoroacetate
步驟1：3-溴-4-氧代哌啶-1-甲酸叔丁酯 Step 1: 3-Bromo-4-oxopiperidine-1-carboxylic acid tert-butyl ester
室溫下，依次將TEA(7.7mL,55mmol)，TMSCl(3.5mL,27.6mmol)加入到4-氧代哌啶-1-甲酸叔丁酯(5g,25mmol)的DMF(30mL)溶液中，混合物75℃攪拌過夜。反應液冷卻到室溫，依次加入冷的飽和碳酸氫鈉水溶液(200mL)和冷的正己烷(200mL)。有機相用飽和食鹽水洗滌，Na2SO4乾燥，濃縮得到粗產品，直接用於下步反應。殘餘物溶解到THF(15mL)中，0℃攪拌15min。緩慢滴加NBS(4.47g,25mmol)的THF(80mL)溶液。滴畢，反應室溫攪拌過夜。將水(200mL)和正己烷(200mL)加入到反應中，有機相用飽和食鹽水洗滌，Na2SO4乾燥，濃縮得到的粗產品，矽膠層析柱(60g矽膠，PE/EA=20/1到8/1梯度)純化，得到白色固體3-溴-4-氧代哌啶-1-甲酸叔丁酯(5.56g,78%)。1H NMR(400MHz,DMSO-d6)δ 4.85-4.70(m,1H),4.20-4.00(m,1H),3.90-3.55(m,3H),2.80-2.68(m,1H),2.54-2.44(m,1H),1.43(s,9H).MS(ESI)m/e[M-t-Bu]+ 221.9,224.0。 TEA (7.7 mL, 55 mmol), TMSCI (3.5 mL, 27.6 mmol) was added to a solution of <RTI ID=0.0>> The mixture was stirred at 75 ° C overnight. The reaction solution was cooled to room temperature, and then cold saturated aqueous sodium hydrogen sulfate (200 mL) and cold n-hexane (200 mL). The organic phase was washed with brine, dried over Na 2 CH 4 The residue was dissolved in THF (15 mL) and stirred 15 min. A solution of NBS (4.47 g, 25 mmol) in THF (80 mL) was slowly evaporated. After the dropwise addition, the reaction was stirred at room temperature overnight. Water (200mL) and hexane (200mL) was added to the reaction, the organic phase was washed with saturated brine, Na 2 SO 4 dried, and concentrated to give the crude product by column chromatography on silica gel (60g silica gel, PE / EA = 20 / Purification by a gradient from 1 to 8/1 afforded a white solid, 3-bromo-4-oxopiperidine-l-carboxylic acid tert-butyl ester (5.56 g, 78%). 1 H NMR (400 MHz, DMSO-d 6 ) δ 4.85-4.70 (m, 1H), 4.20 - 4.00 (m, 1H), 3.90-3.55 (m, 3H), 2.80-2.68 (m, 1H), 2.54- 2.44 (m, 1H), 1.43 (s, 9H). MS (ESI) m/e [Mt-Bu] + 221.9, 224.0.
步驟2：3-氰基-2-(4-苯氧基苯基)-5,6-二氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-7(8H)-甲酸叔丁酯 Step 2: 3-Cyano-2-(4-phenoxyphenyl)-5,6-dihydro-4H-pyrazolo[5',1':2,3]imidazo[4,5- c] Pyridin-7(8H)-tert-butyl formate
5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈(1.5g,5.4mmol)和K2CO3(2.24g,16.3mmol)在DMF(50mL)的混合物，80℃氮氣條件下，攪拌45min。然後，一次性加入3-溴-4-氧代哌啶-1-甲酸叔丁酯(4.5g,16.3mmol)。反應混合物80℃攪拌1小時。然後冷卻到室溫，加入水(150mL)和EA(150mL)。水相進一步用EA(100mL×3)萃取，合併有機相，飽和食鹽水洗滌，Na2SO4乾燥，濃縮，矽膠層析柱(15g矽膠，DCM/MeOH=400/1到200/1)純化，得到類白色固體產品3-氰基-2-(4-苯氧基苯基)-5,6-二氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-7(8H)-甲酸叔丁酯(850mg,35%)。MS(ESI)m/e[M+1]+ 455.9。 5-Amino-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitrile (1.5 g, 5.4 mmol) and K 2 CO 3 (2.24 g, 16.3 mmol) in DMF (50 mL) The mixture was stirred at 80 ° C under nitrogen for 45 min. Then, tert-butyl 3-bromo-4-oxopiperidine-1-carboxylate (4.5 g, 16.3 mmol) was added in one portion. The reaction mixture was stirred at 80 ° C for 1 hour. It was then cooled to room temperature and water (150 mL) and EA (150 mL). The aqueous phase was further extracted with EA (100 mL×3), and the organic phase was combined, washed with saturated brine, dried Na 2 SO 4 , concentrated, and purified by silica gel chromatography (15 g EtOAc, DCM / MeOH = 400/1 to 200/1) To give an off-white solid product 3-cyano-2-(4-phenoxyphenyl)-5,6-dihydro-4H-pyrazolo[5',1':2,3]imidazo[4 , 5-c]pyridine-7(8H)-carboxylic acid tert-butyl ester (850 mg, 35%). MS (ESI) m / e [M+1] + 455.9.
步驟3：2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺三氟乙酸鹽 Step 3: 2-(4-Phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[4,5-c Pyridine-3-carboxamide trifluoroacetate
3-氰基-2-(4-苯氧基苯基)-5,6-二氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-7(8H)-甲酸叔丁酯(130mg,0.28 mmol)的H3PO4(85wt.%水溶液，20mL)溶液在100℃攪拌1.5小時，直到TLC和LCMS監測反應完全。混合物冷卻到室溫，倒入水(100mL)中，用固體K2CO3調節pH到9-10。懸濁液用EA(100mL×4)萃取。合併有機相，飽和食鹽水(200mL)洗滌，Na2SO4乾燥，濃縮得到粗產品。製備HPLC純化，水相含0.1% TFA，10%到90% CH3CN梯度洗脫，收集合併目標產品，凍幹得到白色固體2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺三氟乙酸鹽(15mg,11%)。1H NMR(400MHz,DMSO-d6)δ 11.99(s,1H),9.32(s,2H),7.66(d,J=8.6Hz,2H),7.43(d,J=7.6Hz,1H),7.41(d,J=7.6Hz,1H),7.18(t,J=7.6Hz,1H),7.10-7.06(m,4H),4.44(s,2H),3.49(m,2H),2.95-2.92(m,2H).MS(ESI)m/e[M+1]+ 373.9。 3-cyano-2-(4-phenoxyphenyl)-5,6-dihydro-4H-pyrazolo[5',1':2,3]imidazo[4,5-c]pyridine -7 (8H) - carboxylate (130mg, 0.28 mmol) in H 3 PO 4 (. 85wt% aqueous solution, 20 mL) was stirred at 100 ℃ 1.5 hours until complete reaction was monitored by TLC and LCMS. The mixture was cooled to room temperature, poured into water (100 mL), treated with solid K 2 CO 3 pH was adjusted to 9-10. The suspension was extracted with EA (100 mL x 4). Washed, Na dried organic phases were combined, saturated brine (200mL) 2 SO 4, and concentrated to give the crude product. Purified by preparative HPLC, the aqueous phase was eluted with 0.1% TFA, 10% to 90% CH 3 CN gradient, and the combined product was collected and lyophilized to give 2-(4-phenoxyphenyl)-5,6,7 as a white solid. , 8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[4,5-c]pyridine-3-carboxamide trifluoroacetate (15 mg, 11%). 1 H NMR (400MHz, DMSO- d 6) δ 11.99 (s, 1H), 9.32 (s, 2H), 7.66 (d, J = 8.6Hz, 2H), 7.43 (d, J = 7.6Hz, 1H), 7.41 (d, J = 7.6 Hz, 1H), 7.18 (t, J = 7.6 Hz, 1H), 7.10-7.06 (m, 4H), 4.44 (s, 2H), 3.49 (m, 2H), 2.95-2.92 (m, 2H). MS (ESI) m/e [M+1] + 373.9.
化合物52：7-丙烯醯基-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺 Compound 52: 7-Propanyl-2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazole [4,5-c]pyridine-3-carboxamide
目標產物由化合物51和丙烯醯氯，根據化合物8的類似方法合成。1H NMR(400MHz,DMSO-d6 at 80℃)δ 11.55(s,1H),7.71(d,J=8.6Hz,2H),7.43(d,J=7.6Hz,1H),7.41(d,J=7.6Hz,1H),7.18(t,J=7.6Hz,1H),7.10-7.05(m,4H), 6.88(dd,J=10.6,17.1Hz,1H),6.24(s,2H),6.15(d,J=17.1Hz,1H),5.74(d,J=10.6Hz,1H),4.78(s,2H),3.94-3.91(m,2H),2.80-2.76(m,2H).MS(ESI)m/e[M+1]+ 427.9. The target product was synthesized from Compound 51 and acryloyl chloride in a similar manner to Compound 8 . 1 H NMR (400 MHz, DMSO-d 6 at 80 ° C) δ 11.55 (s, 1H), 7.71 (d, J = 8.6 Hz, 2H), 7.43 (d, J = 7.6 Hz, 1H), 7.41 (d, J = 7.6 Hz, 1H), 7.18 (t, J = 7.6 Hz, 1H), 7.10-7.05 (m, 4H), 6.88 (dd, J = 10.6, 17.1 Hz, 1H), 6.24 (s, 2H), 6.15 (d, J = 17.1 Hz, 1H), 5.74 (d, J = 10.6 Hz, 1H), 4.78 (s, 2H), 3.94 - 3.91 (m, 2H), 2.80 - 2.76 (m, 2H). (ESI)m/e[M+1] + 427.9.
化合物53：7-(3-氯丙醯基)-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺 Compound 53 : 7-(3-Chloropropyl)-2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2 , 3]imidazo[4,5-c]pyridine-3-carboxamide
將2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺三氟乙酸鹽(40mg,0.09mmol)懸浮在氯化氫氣體/二氧六環飽和溶液(50mL)中。混合物室溫攪拌約1.5小時，濃縮。殘餘物懸浮於MeOH(2mL)和水(2mL)中。棄去有機層，水層冷凍乾燥，得到類白色固體7-(3-氯丙醯基)-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺(40mg,90%)。1H NMR(400MHz,DMSO-d6)δ 11.79-11.76(m,1H),7.68(d,J=8.6Hz,2H),7.43(d,J=7.6Hz,1H),7.41(d,J=7.6Hz,1H),7.18(t,J=7.6Hz,1H),7.10-7.04(m,4H),4.71-4.70(m,2H),3.85-3.79(m,4H),3.02(t,J=6.4Hz,2H),2.79-2.69(m,2H).MS(ESI)m/e[M+1]+ 463.8,465.8。 2-(4-Phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[4,5-c]pyridine 3-Mergamine trifluoroacetate (40 mg, 0.09 mmol) was suspended in a hydrogen chloride gas / dioxane saturated solution (50 mL). The mixture was stirred at room temperature for about 1.5 hours and concentrated. The residue was suspended in MeOH (2 mL) and water (2 mL). The organic layer was discarded and the aqueous layer was lyophilized to give an off-white solid 7-(3-chloropropenyl)-2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H- Pyrazolo[5',1':2,3]imidazo[4,5-c]pyridine-3-carboxamide (40 mg, 90%). 1 H NMR (400MHz, DMSO- d 6) δ 11.79-11.76 (m, 1H), 7.68 (d, J = 8.6Hz, 2H), 7.43 (d, J = 7.6Hz, 1H), 7.41 (d, J = 7.6 Hz, 1H), 7.18 (t, J = 7.6 Hz, 1H), 7.10-7.04 (m, 4H), 4.71-4.70 (m, 2H), 3.85-3.79 (m, 4H), 3.02 (t, J = 6.4 Hz, 2H), 2.79-2.69 (m, 2H). MS (ESI) m/e [M+1] + 463.8, 465.8.
化合物54和55：(E)-7-(4-(二甲基氨基)丁-2-烯醯基)-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c] 吡啶-3-甲醯胺和7-乙醯基-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺 Compounds 54 and 55 : ( E )-7-(4-(Dimethylamino)but-2-enyl)-2-(4-phenoxyphenyl)-5,6,7,8-tetra Hydrogen-4H-pyrazolo[5',1':2,3]imidazo[4,5-c]pyridine-3-carboxamide and 7-acetamido-2-(4-phenoxybenzene) -5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[4,5-c]pyridine-3-carboxamide
(E)-4-(二甲基氨基)丁-2-烯酸鹽酸鹽(147mg,0.88mmol)，HATU(611mg,1.6mmol)，TEA(328mg,3.2mmol)在DCM(50ml)中的混合物，室溫下攪拌約2小時，然後加入2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺(300mg,0.8mmol)。混合物室溫攪拌過夜。TLC和LCMS分析表明，初始原料被消耗。向反應溶液中加入水(100mL)和DCM(50mL)。水相進一步用DCM(50ml)萃取。合併有機相，飽和食鹽水洗滌，Na2SO4乾燥，濃縮得到粗產物。矽膠層析柱(5g矽膠，DCM/MeOH=20/1到10/1)純化，冷凍乾燥得到產品(E)-7-(4-(二甲基氨基)丁-2-烯醯基)-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺(145mg,37%)。1H NMR(400MHz,DMSO-d6 at 80℃)δ 11.52(s,1H),7.69(d,J=8.6Hz,2H),7.42(d,J=7.6Hz,1H),7.41(d,J=7.6Hz,1H),7.16(t,J=7.6Hz,1H),7.12-6.99(m,4H),6.78-6.56(m,2H),6.22(s,2H),4.75(s,2H), 3.89(t,J=5.6Hz,2H),3.12(d,J=5.6Hz,2H),2.80-2.72(m,2H),2.22(s,6H).MS(ESI)m/e[M+1]+ 484.9。 ( E )-4-(Dimethylamino)but-2-enoate (147 mg, 0.88 mmol), HATU (611 mg, 1.6 mmol), TEA (328 mg, 3.2 mmol) in DCM (50 ml) The mixture was stirred at room temperature for about 2 hours, then 2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazole [5',1':2,3 was added. Imidazo[4,5-c]pyridine-3-carboxamide (300 mg, 0.8 mmol). The mixture was stirred at room temperature overnight. TLC and LCMS analysis indicated that the starting material was consumed. Water (100 mL) and DCM (50 mL) were added to the reaction mixture. The aqueous phase was further extracted with DCM (50 mL). The combined organic phases were washed with brine, dried Na 2 SO 4, and concentrated to give the crude product. Purification by a silica gel chromatography column (5 g of hydrazine, DCM/MeOH = 20/1 to 10/1), lyophilized to give the product ( E )-7-(4-(dimethylamino)but-2-enyl)- 2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[4,5-c]pyridine- 3-Protonamine (145 mg, 37%). 1 H NMR (400 MHz, DMSO-d 6 at 80 ° C) δ 11.52 (s, 1H), 7.69 (d, J = 8.6 Hz, 2H), 7.42 (d, J = 7.6 Hz, 1H), 7.41 (d, J = 7.6 Hz, 1H), 7.16 (t, J = 7.6 Hz, 1H), 7.12-6.99 (m, 4H), 6.78-6.56 (m, 2H), 6.22 (s, 2H), 4.75 (s, 2H) ), 3.89 (t, J = 5.6 Hz, 2H), 3.12 (d, J = 5.6 Hz, 2H), 2.80-2.72 (m, 2H), 2.22 (s, 6H). MS (ESI) m/e [ M+1] + 484.9.
因為上步中殘餘的HOAc，得到副產品7-乙醯基-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺。1H NMR(400MHz,DMSO-d6)δ 11.77-11.73(m,1H),7.70-7.66(m,2H),7.43(d,J=7.6Hz,1H),7.41(d,J=7.6Hz,1H),7.18(t,J=7.6Hz,1H),7.13-6.94(m,4H),4.68(s,2H),3.83-3.76(m,2H),2.82-2.77(m,2H),2.14(s,3H).MS(ESI)m/e[M+1]+ 416。 Because of the residual HOAc in the previous step, the by-product 7-acetamido-2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazole[5',1' was obtained. : 2,3]Imidazo[4,5-c]pyridine-3-carbamide. 1 H NMR (400MHz, DMSO- d 6) δ 11.77-11.73 (m, 1H), 7.70-7.66 (m, 2H), 7.43 (d, J = 7.6Hz, 1H), 7.41 (d, J = 7.6Hz , 1H), 7.18 (t, J = 7.6 Hz, 1H), 7.13-6.94 (m, 4H), 4.68 (s, 2H), 3.83-3.76 (m, 2H), 2.82-2.77 (m, 2H), 2.14 (s, 3H). MS (ESI) m / e [M + 1] + 416.
化合物56：7-(2-氰基乙醯基)-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺 Compound 56 : 7-(2-Cyanoethenyl)-2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1': 2,3]imidazo[4,5-c]pyridine-3-carboxamide
目標產品是由2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺和氰乙酸，根據化合物54的類似方法合成。1H NMR(400MHz,DMSO-d6 at 80℃)δ 11.55(s,1H),7.68(d,J=8.6Hz,2H),7.41(d,J=7.6Hz,1H),7.39(d,J=7.6Hz,1H),7.16(t,J=7.6Hz,1H),7.11-6.93(m,4H),6.22(br s,2H),4.68(s,2H),4.14(s,2H),3.86- 3.79(m,2H),2.84-2.73(m,2H).MS(ESI)m/e[M+1]+ 440.9。 The target product is 2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[4,5- c] Pyridine-3-carboamine and cyanoacetic acid, synthesized according to a similar method of compound 54 . 1 H NMR (400 MHz, DMSO-d 6 at 80 ° C) δ 11.55 (s, 1H), 7.68 (d, J = 8.6 Hz, 2H), 7.41 (d, J = 7.6 Hz, 1H), 7.39 (d, J = 7.6 Hz, 1H), 7.16 (t, J = 7.6 Hz, 1H), 7.11-6.93 (m, 4H), 6.22 (br s, 2H), 4.68 (s, 2H), 4.14 (s, 2H) , 3.86- 3.79 (m, 2H), 2.84-2.73 (m, 2H). MS (ESI) m/e [M+1] + 440.9.
化合物57：7-(3-(二甲基氨基)丙醯基)-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺三氟乙酸鹽 Compound 57 : 7-(3-(Dimethylamino)propanyl)-2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[4,5-c]pyridine-3-carboxamide trifluoroacetate
向7-丙烯醯基-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺(6mg,0.014mmol)的MeOH(5mL)溶液中，室溫下依次加入NaOMe(15mg,0.28mmol)，二甲胺鹽酸鹽(12mg,014mmol)。混合物50℃攪拌過夜。冷卻至室溫後，混合物濃縮。殘餘物通過製備HPLC純化，0%到60%CH3CN在H2O中梯度洗脫。收集合併目標產品，過夜凍幹，得到黃色固體7-(3-(二甲基氨基)丙醯基)-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺三氟乙酸鹽(2.5mg,35%)。1H NMR(400MHz,DMSO-d6)δ 11.84-11.82(m,1H),9.55(s,1H),7.69-7.66(m,2H),7.44(d,J=7.6Hz,1H),7.42(d,J=7.6Hz,1H),7.18(t,J=7.6Hz,1H),7.10-7.04(m,4H),4.75-4.72(m,2H),3.87-3.82(m, 2H),3.02-3.00(m,2H),2.84-2.78(m,2H),2.77(s,6H),2.71-2.68(m,2H).MS(ESI)m/e[M+1]+ 472.9。 To 7-propenyl-2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[4 ,5-c]pyridine-3-carboxamide (6 mg, 0.014 mmol) in MeOH (5 mL) EtOAc (EtOAc) . The mixture was stirred at 50 ° C overnight. After cooling to room temperature, the mixture was concentrated. The residue was purified by preparative HPLC, 0% to 60% CH 3 CN in H 2 O gradient of. The combined target product was collected and lyophilized overnight to give 7-(3-(dimethylamino)propanyl)-2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro as a yellow solid. -4H-pyrazolo[5',1':2,3]imidazo[4,5-c]pyridine-3-carboxamide trifluoroacetate (2.5 mg, 35%). 1 H NMR (400MHz, DMSO- d 6) δ 11.84-11.82 (m, 1H), 9.55 (s, 1H), 7.69-7.66 (m, 2H), 7.44 (d, J = 7.6Hz, 1H), 7.42 (d, J = 7.6 Hz, 1H), 7.18 (t, J = 7.6 Hz, 1H), 7.10-7.04 (m, 4H), 4.75-4.72 (m, 2H), 3.87-3.82 (m, 2H), 3.02-3.00 (m, 2H), 2.84-2.78 (m, 2H), 2.77 (s, 6H), 2.71-2.68 (m, 2H). MS (ESI) m/e [M+1] + 472.9.
化合物58：7-(丁基-2-烯醯基)-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺 Compound 58 : 7-(butyl-2-enyl)-2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1' :2,3]Imidazo[4,5-c]pyridine-3-carboxamide
目標產品是由2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺和巴豆酸，根據化合物54的類似方法合成。1H NMR(400MHz,DMSO-d6)δ 11.78-11.71(m,1H),7.68(d,J=8.6Hz,2H),7.46-7.39(m,2H),7.18(t,J=7.6Hz,1H),7.11-7.04(m,4H),6.73-6.63(m,2H),4.80-4.71(m,2H),3.90(s,2H),2.76-2.70(m,2H),1.88-1.86(m,3H).MS(ESI)m/e[M+1]+ 441.9。 The target product is 2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[4,5- c] Pyridine-3-carboamine and crotonic acid were synthesized according to a similar method of compound 54 . 1 H NMR (400MHz, DMSO- d 6) δ 11.78-11.71 (m, 1H), 7.68 (d, J = 8.6Hz, 2H), 7.46-7.39 (m, 2H), 7.18 (t, J = 7.6Hz , 1H), 7.11-7.04 (m, 4H), 6.73-6.63 (m, 2H), 4.80-4.71 (m, 2H), 3.90 (s, 2H), 2.76-2.70 (m, 2H), 1.88-1.86 (m, 3H). MS (ESI) m/e [M+1] + 441.9.
化合物59和60：(E)-7-(3-腈基丁烯基)-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺和(Z)-7-(3-腈基丁烯基)-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺 Compounds 59 and 60 : ( E )-7-(3-nitrilebutenyl)-2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[ 5',1':2,3]imidazo[4,5-c]pyridine-3-carboxamide and ( Z )-7-(3-cyanobutenyl)-2-(4-phenoxy) Phenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[4,5-c]pyridine-3-carboxamide
室溫下，向2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺(100mg,0.268mmol)的丙酮(10mL)溶液中，加入K2CO3(140mg,1.07mmol)。室溫攪拌2小時後，加入4-溴丁基-2-烯腈(40mg,0.268mmol)的2mL丙酮溶液，室溫攪拌過夜。然後，混合物在EA(50mL)和水(100mL)中分配。水相進一步用EA(50mL)萃取，合併有機相，食鹽水洗滌，Na2SO4乾燥，濃縮，得到粗產物，進一步製備矽膠板(DCM/MeOH=15/1)純化，得到淺黃色固體(E)-7-(3-腈基丁烯基)-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺(7mg,6%)。1H NMR(400MHz,CDCl3)δ 10.45(s,1H),7.59(d,J=8.4Hz,2H),7.42-7.30(m,2H),7.16(t,J=7.6Hz,1H),7.10(d,J=8.4Hz,2H),7.06(d,J=7.6Hz,2H),6.76(dt,J=16.3,4.6Hz,1H),5.67(d,J=16.3Hz,1H),5.62(s,2H),3.82(s,2H),3.41(d,J=4.6Hz,2H),2.95-2.75(m,4H).MS(ESI)m/e[M+1]+ 439.9。 To 2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[4,5 at room temperature -C] pyridine-3-acyl-amine (100mg, 0.268mmol) in acetone (10 mL) was added K 2 CO 3 (140mg, 1.07mmol ). After stirring at room temperature for 2 hours, a solution of 4-bromobutyl-2-enenitrile (40 mg, 0.268 mmol) in 2 mL of acetone was added and stirred at room temperature overnight. The mixture was then partitioned between EA (50 mL) and water (100 mL). The aqueous phase was further extracted with EA (50mL). The combined organic phases were washed with brine, dried Na 2 SO 4, and concentrated to give the crude product which was purified further preparation silica gel plate (DCM / MeOH = 15/1 ), to give a pale yellow solid ( E )-7-(3-cyanobutenyl)-2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1': 2,3] Imidazo[4,5-c]pyridine-3-carbamide (7 mg, 6%). 1 H NMR (400MHz, CDCl 3 ) δ 10.45 (s, 1H), 7.59 (d, J = 8.4Hz, 2H), 7.42-7.30 (m, 2H), 7.16 (t, J = 7.6Hz, 1H), 7.10 (d, J = 8.4 Hz, 2H), 7.06 (d, J = 7.6 Hz, 2H), 6.76 (dt, J = 16.3, 4.6 Hz, 1H), 5.67 (d, J = 16.3 Hz, 1H), 5.62 (s, 2H), 3.82 (s, 2H), 3.41 (d, J = 4.6 Hz, 2H), 2.95 - 2.75 (m, 4H). MS (ESI) m/e [M+1] + 439.9.
淺黃色固體(Z)-7-(3-腈基丁烯基)-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺(4mg,3.4%)。1H NMR(400MHz,CDCl3)δ 10.22(s,1H),7.60(d,J=8.4Hz,2H),7.41-7.33(m,2H),7.16(t,J=7.6Hz,1H),7.10(d,J=8.4Hz,2H),7.06(d,J=7.6Hz,2H),6.70-6.59(m,1H),5.58(s,2H),5.53(d,J=11.2Hz,1H),3.85(s, 2H),3.64(d,J=6.4Hz,2H),2.99-2.80(m,4H).MS(ESI)m/e[M+1]+ 439.9。 Light yellow solid ( Z )-7-(3-cyanobutenyl)-2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5' , 1': 2,3]imidazo[4,5-c]pyridine-3-carbamide (4 mg, 3.4%). 1 H NMR (400MHz, CDCl 3 ) δ 10.22 (s, 1H), 7.60 (d, J = 8.4Hz, 2H), 7.41-7.33 (m, 2H), 7.16 (t, J = 7.6Hz, 1H), 7.10 (d, J = 8.4 Hz, 2H), 7.06 (d, J = 7.6 Hz, 2H), 6.70-6.59 (m, 1H), 5.58 (s, 2H), 5.53 (d, J = 11.2 Hz, 1H) ), 3.85 (s, 2H), 3.64 (d, J = 6.4 Hz, 2H), 2.99-2.80 (m, 4H). MS (ESI) m/e [M+1] + 439.9.
實施例22：化合物61-64的合成Example 22: Synthesis of compound 61-64
化合物61：2-(4-苯氧基苯基)-4H-吡唑並[1',5'：1,2]咪唑並[4,5-c]吡啶-3-甲醯胺 Compound 61 : 2-(4-phenoxyphenyl)-4H-pyrazolo[1',5':1,2]imidazo[4,5-c]pyridine-3-carboxamide
步驟1：3-溴-4-肼基吡啶 Step 1: 3-bromo-4-indolylpyridine
3-溴-4-氯吡啶(5g,0.026mol)和水合肼(80%水溶液，80mL)在二氧六環(100mL)中的混合物，100℃攪拌過夜。冷卻至室溫，混合物濃縮。殘餘物在EA(300mL)和飽和氯化銨溶液(300mL)中分配。有機相鹽水洗滌，Na2SO4乾燥，過濾，濃縮得到粗產物，懸浮于冷的異丙醇(30mL)中，過濾。收集的固體在空氣中乾燥，得到白色固體3-溴-4-肼基吡啶(4.2g,87%)。1H NMR(400MHz,DMSO-d6)δ 8.18(s,1H),8.07(d,J=5.6Hz,1H),7.37(s,1H),7.01(d,J=5.6Hz,1H),4.36(s,2H)。 A mixture of 3-bromo-4-chloropyridine (5 g, 0.026 mol) and hydrazine hydrate (80% aqueous solution, 80 mL) in dioxane (100 mL) was stirred at 100 ° C overnight. Cool to room temperature and concentrate the mixture. The residue was partitioned between EA (300 mL) andEtOAc. The organic phase was washed with brine, Na dried 2 SO 4, filtered, and concentrated to give the crude product was suspended in cold isopropanol (30mL) and filtered. The collected solid was dried in air to give 3-bromo-4-mercaptopyridine (4.2 g, 87%). 1 H NMR (400MHz, DMSO- d 6) δ 8.18 (s, 1H), 8.07 (d, J = 5.6Hz, 1H), 7.37 (s, 1H), 7.01 (d, J = 5.6Hz, 1H), 4.36 (s, 2H).
步驟2：5-氨基-1-(3-溴吡啶-4-基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈 Step 2: 5-Amino-1-(3-bromopyridin-4-yl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitrile
2-(甲氧基(4-苯氧基苯基)亞甲基)丙二腈(7.3g,0.026mol)和3-溴-4-肼基吡啶(4.2g,0.022mol)在乙醇(300mL)中的混合物，氮氣保護下回流過夜。反應液緩慢冷卻至室溫，室溫攪拌約4小時，直到產生固體沉澱。將固體物過濾，收集，正己烷洗滌得到淺黃色固體5-氨基-1-(3-溴吡啶-4-基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈(3.38g,35%)。MS(ESI)m/e[M+1]+ 431.8,433.8。 2-(Methoxy(4-phenoxyphenyl)methylene)malononitrile (7.3 g, 0.026 mol) and 3-bromo-4-indolylpyridine (4.2 g, 0.022 mol) in ethanol (300 mL) The mixture was refluxed overnight under nitrogen. The reaction solution was slowly cooled to room temperature and stirred at room temperature for about 4 hours until a solid precipitate formed. The solid was filtered, collected and washed with n-hexane to afford 5-amino-1-(3-bromopyridin-4-yl)-3-(4-phenoxyphenyl)-1H-pyrazole-4- Formonitrile (3.38 g, 35%). MS (ESI) m / e [ M + 1] + 431.8,433.8.
步驟3：5-氨基-1-(3-溴吡啶-4-基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺 Step 3: 5-Amino-1-(3-bromopyridin-4-yl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide
目標產品是由5-氨基-1-(3-溴吡啶-4-基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈根據化合物51的類似方法合成。MS(ESI)m/e[M+1]+ 449.8,451.8。 The objective product was synthesized by a similar method of compound 51 from 5-amino-1-(3-bromopyridin-4-yl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitrile. MS (ESI) m / e [ M + 1] + 449.8,451.8.
步驟4：2-(4-苯氧基苯基)-4H-吡唑並[1',5'：1,2]咪唑並[4,5-c]吡啶-3-甲醯胺 Step 4: 2-(4-Phenoxyphenyl)-4H-pyrazolo[1',5':1,2]imidazo[4,5-c]pyridine-3-carboxamide
5-氨基-1-(3-溴吡啶-4-基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺(3.96g,8.8mmol),CuI(836mg,4.4mmol),N 1,N 2-二甲基乙基-1,2-二胺(77mg,0.88mmol)和K3PO4(5.59g,26.4mmol)在DMF(100mL)中的混合物，氮氣保護下，100℃攪拌2小時，直至TLC表明大部分起始原料被消耗。冷卻至室溫後，混合物過濾，濃縮。殘餘物用矽膠層析柱(30g矽膠，DCM/MeOH=20/1到10/1洗脫)純化，得到褐色固體2-(4-苯氧基苯基)-4H-吡唑並[1',5'：1,2]咪唑並[4,5-c]吡啶-3-甲醯胺(3.2g,99%)。1H NMR(400MHz,DMSO-d6)δ 12.63(br s,1H),8.85(s,1H),8.46(s,1H),7.96(d,J=4.6Hz,1H),7.81(d,J=8.6Hz,2H),7.45(d,J=7.6Hz,1H),7.43(d,J=7.6Hz,1H),7.20(t,J=7.6Hz,1H),7.14-7.09(m,4H).MS(ESI)m/e[M+1]+ 369.9。 5-amino-1-(3-bromopyridin-4-yl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide (3.96 g, 8.8 mmol), CuI (836 mg) a mixture of 4.4 mmol), N 1 , N 2 -dimethylethyl-1,2-diamine (77 mg, 0.88 mmol) and K 3 PO 4 (5.59 g, 26.4 mmol) in DMF (100 mL) The mixture was stirred at 100 ° C for 2 hours under nitrogen atmosphere until TLC indicated that most of the starting material was consumed. After cooling to room temperature, the mixture was filtered and concentrated. The residue was purified with a silica gel chromatography eluting eluting eluting eluting eluting , 5': 1, 2] imidazo[4,5-c]pyridine-3-carboxamide (3.2 g, 99%). 1 H NMR (400MHz, DMSO- d 6) δ 12.63 (br s, 1H), 8.85 (s, 1H), 8.46 (s, 1H), 7.96 (d, J = 4.6Hz, 1H), 7.81 (d, J = 8.6 Hz, 2H), 7.45 (d, J = 7.6 Hz, 1H), 7.43 (d, J = 7.6 Hz, 1H), 7.20 (t, J = 7.6 Hz, 1H), 7.14 - 7.09 (m, 4H). MS (ESI) m/e [M+1] + 369.9.
化合物62：2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[1',5'：1,2]咪唑並[4,5-c]吡啶-3-甲醯胺 Compound 62 : 2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[1',5':1,2]imidazo[4,5-c Pyridine-3-carboxamide
步驟1：6-苄基-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺 Step 1: 6-Benzyl-2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[ 4,5-c]pyridine-3-carboxamide
向2-(4-苯氧基苯基)-4H-吡唑並[1',5'：1,2]咪唑並[4,5-c]吡啶-3-甲醯胺(2.46g,0.0067mol)的THF(150mL)懸濁液中，緩慢滴加苄溴(1.14g,0.0067mol)。然後，混合物65℃攪拌過夜。冷卻至室溫後，混合物濃縮。殘餘物懸浮於MeOH(150mL)中，分批加入NaBH4(10g,0.26mol)，混合物室溫攪拌過夜。向反應溶液中依次加入水(200mL)，DCM(200mL)。水相進一步用DCM(100mL)萃取。合併有機相，飽和食鹽水洗滌，Na2SO4乾燥，濃縮，得到粗產品，殘餘物用矽膠層析柱(10g矽膠，DCM/MeOH=200/1到80/1洗脫)純化，得到褐色泡沫狀物6-苄基-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺(0.786g,26%)。MS(ESI)m/e[M+1]+ 463.9。 To 2-(4-phenoxyphenyl)-4H-pyrazolo[1',5':1,2]imidazo[4,5-c]pyridine-3-carboxamide (2.46 g, 0.0067 In a suspension of THF (150 mL), benzyl bromide (1.14 g, 0.0067 mol) was slowly added dropwise. Then, the mixture was stirred at 65 ° C overnight. After cooling to room temperature, the mixture was concentrated. The residue was suspended in MeOH (150mL), was added portionwise NaBH 4 (10g, 0.26mol), the mixture was stirred at room temperature overnight. Water (200 mL) and DCM (200 mL) were sequentially added to the mixture. The aqueous phase was further extracted with DCM (100 mL). The combined organic phases were washed with brine, dried Na 2 SO 4, and concentrated to give the crude product residue (10g silica gel elution, DCM / MeOH = 200/1 to 80/1) was purified by silica gel column chromatography to give a brown 6-Benzyl-2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[6] 4,5-c]pyridine-3-carbamide (0.786 g, 26%). MS (ESI) m/e [M+1] + 463.9.
步驟2：2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺 Step 2: 2-(4-Phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[4,5-c Pyridine-3-carboxamide
6-苄基-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺(640mg,0.00138mol)和10% w/w Pd/C(700mg)在MeOH(60mL)中的混合物，室溫一個大氣壓H2下攪拌過夜。TLC和LCMS分析表明起始原料被消耗。過濾，濾液濃縮，得到2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺(397mg,77%)。 1H NMR(400MHz,CD3OD-d4)δ 7.65-7.55(m,2H),7.43-7.33(m,2H),7.18-7.12(m,1H),7.11-6.99(m,4H),4.18(s,2H),3.44(t,J=5.8Hz,2H),3.01(t,J=5.8Hz,2H).MS(ESI)m/e[M+1]+ 373.9。 6-Benzyl-2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[4,5 -C] mixture of pyridine-3-acyl-amine (640mg, 0.00138mol) and 10% w / w Pd / C (700mg) in MeOH (60mL), stirred at room temperature under a H 2 atmosphere overnight. Analysis by TLC and LCMS indicated the starting material was consumed. Filtration and concentration of the filtrate gave 2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[4, 5-c]pyridine-3-carbamide (397 mg, 77%). 1 H NMR (400MHz, CD 3 OD-d 4) δ 7.65-7.55 (m, 2H), 7.43-7.33 (m, 2H), 7.18-7.12 (m, 1H), 7.11-6.99 (m, 4H), 4.18 (s, 2H), 3.44 (t, J = 5.8 Hz, 2H), 3.01 (t, J = 5.8 Hz, 2H). MS (ESI) m/e [M+1] + 373.9.
化合物63：6-丙烯醯基-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺三氟乙酸鹽 Compound 63 : 6-propenyl-2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazole [4,5-c]pyridine-3-carboxamide trifluoroacetate
目標產品是由2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺和丙烯醯氯，根據化合物8的類似方法合成。1H NMR(400MHz,DMSO-d6)δ 11.79(s, 1H),7.73(d,J=8.6Hz,2H),7.50(d,J=7.6Hz,1H),7.48(d,J=7.6Hz,1H),7.24(t,J=7.6Hz,1H),7.17-7.11(m,4H),7.06-6.98(m,1H),7.22-6.26(m,1H),5.84-5.82(m,1H),4.72(s,2H),4.00-3.97(m,2H),2.94-2.90(m,2H).MS(ESI)m/e[M+1]+ 427.9。 The target product is 2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[4,5- c] Pyridine-3-carboamine and acrylonitrile chloride were synthesized according to a similar method of Compound 8 . 1 H NMR (400MHz, DMSO- d 6) δ 11.79 (s, 1H), 7.73 (d, J = 8.6Hz, 2H), 7.50 (d, J = 7.6Hz, 1H), 7.48 (d, J = 7.6 Hz, 1H), 7.24 (t, J = 7.6 Hz, 1H), 7.17-7.11 (m, 4H), 7.06-6.98 (m, 1H), 7.22-6.26 (m, 1H), 5.84-5.82 (m, 1H), 4.72 (s, 2H), 4.40 - 3.97 (m, 2H), 2.94 - 2.90 (m, 2H). MS (ESI) m/e [M+1] + 427.9.
化合物64：(E)-6-(4-(二甲基氨基)丁-2-烯醯基)-2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺三氟乙酸鹽 Compound 64 : ( E )-6-(4-(Dimethylamino)but-2-enyl)-2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro- 4H-pyrazolo[5',1':2,3]imidazo[4,5-c]pyridine-3-carboxamide trifluoroacetate
目標產品是由2-(4-苯氧基苯基)-5,6,7,8-四氫-4H-吡唑並[5',1'：2,3]咪唑並[4,5-c]吡啶-3-甲醯胺和反式-4-二甲基胺基巴豆酸鹽酸鹽，根據化合物54的類似方法合成。1H NMR(400MHz,DMSO-d6)δ 11.82-11.74(m,1H),10.06(br s,1H),7.66(d,J=8.6Hz,2H),7.43(d,J=7.6Hz,1H),7.42(d,J=7.6Hz,1H),7.18(t,J=7.6Hz,1H),7.10-7.05(m,4H),6.74-6.54(m,2H),4.66(s,2H),3.99-3.86(m,4H),2.88-2.76(m,2H),2.78(s,6H).MS(ESI)m/e[M+1]+ 484.9。 The target product is 2-(4-phenoxyphenyl)-5,6,7,8-tetrahydro-4H-pyrazolo[5',1':2,3]imidazo[4,5- c] Pyridine-3-carboamine and trans-4-dimethylamino crotonate were synthesized according to a similar procedure of compound 54 . 1 H NMR (400MHz, DMSO- d 6) δ 11.82-11.74 (m, 1H), 10.06 (br s, 1H), 7.66 (d, J = 8.6Hz, 2H), 7.43 (d, J = 7.6Hz, 1H), 7.42 (d, J = 7.6 Hz, 1H), 7.18 (t, J = 7.6 Hz, 1H), 7.10-7.05 (m, 4H), 6.74-6.54 (m, 2H), 4.66 (s, 2H) ), 3.99-3.86 (m, 4H), 2.88-2.76 (m, 2H), 2.78 (s, 6H). MS (ESI) m/e [M+1] + 484.9.
實施例23：化合物65-67的合成Example 23: Synthesis of Compound 65-67
化合物65：7-硝基-2-(4-苯氧基苯基)-4H-苯並[4,5]咪唑並[1,2-b]吡唑-3-甲醯胺 Compound 65 : 7-nitro-2-(4-phenoxyphenyl)-4H-benzo[4,5]imidazo[1,2-b]pyrazole-3-carboxamide
步驟1：(2-溴-5-硝基苯基)肼 Step 1: (2-Bromo-5-nitrophenyl)indole
向2-溴-5-硝基苯胺(1g,4.6mmol)在conc.HCl(10mL)的懸濁液中，0℃緩慢加入NaNO2(382mg,5.5mmol)的水(1.5mL)溶液。混合物0℃攪拌3小時，至TLC和LCMS監測大部分原料反應完全。將SnCl2(1.90g,10mmol)在conc.HCl(3mL)的溶液緩慢加入。之後，混合物室溫攪拌2小時，反應冷卻到0℃。用飽和的碳酸氫鈉溶液調節pH值到7-8。混合物乙酸乙酯(50mL×3)萃取，合併有機相，鹽水洗滌，Na2SO4乾燥，過濾，濃縮得到粗產品。進一步矽膠層析柱(10g矽膠，PE/EA=20/1到4/1洗脫)純化，得到橘紅色固體(2-溴-5-硝基苯基)肼(560mg,51%)。1H NMR(400MHz,DMSO-d6)δ 7.89(d,J=2.8Hz,1H),7.60(d,J=8.6Hz,1H),7.29(dd,J=2.8,8.6Hz,1H),7.04(s,1H),4.38(s,2H).MS(ESI)m/e[M+1]+ 232,234。 2-bromo-5-nitroaniline (1g, 4.6mmol) in conc. HCl (10mL) suspension in, 0 ℃ was slowly added NaNO 2 (382mg, 5.5mmol) in water (1.5mL) was added. The mixture was stirred at 0 ° C for 3 hours, and most of the starting materials were taken to complete by TLC and LCMS. A solution of SnCl 2 (1.90 g, 10 mmol) in conc. HCl (3 mL) was slowly added. Thereafter, the mixture was stirred at room temperature for 2 hours, and the reaction was cooled to 0 °C. The pH was adjusted to 7-8 with saturated sodium bicarbonate solution. Ethyl acetate mixture (50mL × 3). The combined organic phases were washed with brine, dried Na 2 SO 4, filtered, and concentrated to give the crude product. Further, a silica gel column (10 g of phthalocyanine, eluted with PE/EA = 20/1 to 4/1) was purified to give an orange-yellow solid (2-bromo-5-nitrophenyl)indole (560 mg, 51%). 1 H NMR (400MHz, DMSO- d 6) δ 7.89 (d, J = 2.8Hz, 1H), 7.60 (d, J = 8.6Hz, 1H), 7.29 (dd, J = 2.8,8.6Hz, 1H), 7.04 (s, 1H), 4.38 (s, 2H). MS (ESI) m/e [M+1] + 232, 234.
步驟2：5-氨基-1-(2-溴-5-硝基苯基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈 Step 2: 5-Amino-1-(2-bromo-5-nitrophenyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitrile
向2-(甲氧基(4-苯氧基苯基)亞甲基)丙二腈(392mg,1.42mmol)的乙醇(30mL)中，一次性加入(2-溴-5-硝基苯基)肼(300mg,1.29mmol)。之後，混合物70℃氮氣保護下攪拌過夜。將該混合物濃縮至幹，矽膠層析柱(5g矽膠，PE/EA=10/1到2/1洗脫)純化，得到黃色固體5-氨基-1-(2-溴-5-硝基苯基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈(128mg,21%)。MS(ESI)m/e[M+1]+ 476,478。 To 2-(methoxy(4-phenoxyphenyl)methylene)malononitrile (392 mg, 1.42 mmol) in ethanol (30 mL), (2-bromo-5-nitrophenyl) ) 肼 (300 mg, 1.29 mmol). Thereafter, the mixture was stirred at 70 ° C under nitrogen atmosphere overnight. The mixture was concentrated to dryness and purified on a silica gel column (5 g silica gel eluting with PE/EA = 10/1 to 2/1) to give 5-amino-1-(2-bromo-5-nitrobenzene) as a yellow solid. 3-(4-Phenoxyphenyl)-1H-pyrazole-4-carbonitrile (128 mg, 21%). MS (ESI) m/e [M+1] + 476, 478.
步驟3：5-氨基-1-(2-溴-5-硝基苯基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺 Step 3: 5-Amino-1-(2-bromo-5-nitrophenyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide
5-氨基-1-(2-溴-5-硝基苯基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈(137mg,0.287mmol)在磷酸(85wt.% in H2O,10mL)中的混合物，100℃攪拌1小時，直至TLC和LCMS分析表明大部分起始原料反應完全。反應冷卻至室溫，在水(40mL)和EA(40mL)中分配，分離出有機相，水相用EA(20mL)萃取。合併 有機相，飽和食鹽水(50mL)洗滌，Na2SO4乾燥，濃縮，得到粗產物(149mg)，直接用於下一步反應未經進一步純化。MS(ESI)m/e[M+1]+ 494,496。 5-amino-1-(2-bromo-5-nitrophenyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitrile (137 mg, 0.287 mmol) in phosphoric acid (85 wt The mixture in .% in H 2 O, 10 mL) was stirred at 100 ° C for 1 hour until TLC and LCMS analysis indicated that most of the starting material was completely reacted. The reaction was cooled to rt. EtOAc (EtOAc)EtOAc. Washed, Na dried organic phases were combined, saturated brine (50mL) 2 SO 4, and concentrated to give the crude product (149 mg), without further purification was used directly in the next reaction. MS (ESI) m / e [M+1] + 494, 496.
步驟3：7-硝基-2-(4-苯氧基苯基)-4H-苯並[4,5]咪唑並[1,2-b]吡唑-3-甲醯胺 Step 3: 7-Nitro-2-(4-phenoxyphenyl)-4H-benzo[4,5]imidazo[1,2-b]pyrazole-3-carboxamide
5-氨基-1-(2-溴-5-硝基苯基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺(149mg,0.3mmol,crude)，CuI(5.7mg,0.03mmol)，N 1,N 2-二甲基乙基-1,2-二胺(3mg,0.03mmol)，K3PO4(64mg,0.3mol)在DMF(15mL)中的混合物，氮氣保護下60℃攪拌5小時，直至TLC分析表明大部分起始原料被消耗。反應冷卻至室溫，減壓去除溶劑。殘餘物矽膠層析柱(5g矽膠，DCM/MeOH=200/1到20/1洗脫)純化，得到褐色固體7-硝基-2-(4-苯氧基苯基)-4H-苯並[4,5]咪唑並[1,2-b]吡唑-3-甲醯胺(62mg,52%)。MS(ESI)m/e[M+1]+ 414。 5-amino-1-(2-bromo-5-nitrophenyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide (149 mg, 0.3 mmol, crude), CuI (5.7 mg, 0.03 mmol), N 1 , N 2 -dimethylethyl-1,2-diamine (3 mg, 0.03 mmol), K 3 PO 4 (64 mg, 0.3 mol) in DMF (15 mL) The mixture was stirred at 60 ° C for 5 hours under nitrogen atmosphere until TLC analysis indicated that most of the starting material was consumed. The reaction was cooled to room temperature and the solvent was removed under reduced pressure. The residue was chromatographed (5 g EtOAc (EtOAc) elute elut elut elut elut elut elut elut [4,5] Imidazo[1,2-b]pyrazole-3-carboxamide (62 mg, 52%). MS (ESI) m/e [M+1] + 414.
化合物66：7-氨基-2-(4-苯氧基苯基)-4H-苯並[4,5]咪唑並[1,2-b]吡唑-3-甲醯胺 Compound 66 : 7-Amino-2-(4-phenoxyphenyl)-4H-benzo[4,5]imidazo[1,2-b]pyrazole-3-carboxamide
向7-硝基-2-(4-苯氧基苯基)-4H-苯並[4,5]咪唑並[1,2-b]吡唑-3-甲醯胺(9mg,0.022mmol)的HOAc(3mL)中，加入鋅粉(14mg,0.22mmol)。混合物室溫攪拌20分鐘，直到TLC和LCMS分析表明大部分的起始原料被消耗。將反應固體濾出，濾液濃縮，殘餘物懸浮於EA(10mL)中，過濾。將濾液濃縮，得到白色固體產品(4mg,50%)。1H NMR(400MHz,CD3OD-d4)δ 7.60(d,J=8.6Hz,2H),7.32(d,J=8.0Hz,1H),7.30(d,J=8.0Hz,1H),7.23(d,J=8.4Hz,1H),7.16(s,1H),7.08(t,J=8.0Hz,1H),7.05-6.95(m,4H),6.79(dd,J=1.6,8.4Hz,1H).MS(ESI)m/e[M+1]+ 384。 To 7-nitro-2-(4-phenoxyphenyl)-4H-benzo[4,5]imidazo[1,2-b]pyrazole-3-carboxamide (9 mg, 0.022 mmol) Zinc powder (14 mg, 0.22 mmol) was added to HOAc (3 mL). The mixture was stirred at room temperature for 20 minutes until TLC and LCMS analysis indicated that most of the starting material was consumed. The reaction solid was filtered, and the filtrate was evaporated. The filtrate was concentrated to give a white solid (4mg, 50%). 1 H NMR (400 MHz, CD 3 OD-d 4 ) δ 7.60 (d, J = 8.6 Hz, 2H), 7.32 (d, J = 8.0 Hz, 1H), 7.30 (d, J = 8.0 Hz, 1H), 7.23 (d, J = 8.4 Hz, 1H), 7.16 (s, 1H), 7.08 (t, J = 8.0 Hz, 1H), 7.05-6.95 (m, 4H), 6.79 (dd, J = 1.6, 8.4 Hz) , 1H). MS (ESI) m / e [M + 1] + 384.
化合物67：7-丙烯醯胺基-2-(4-苯氧基苯基)-4H-苯並[4,5]咪唑並[1,2-b]吡唑-3-甲醯胺 Compound 67 : 7-Acrylamino-2-(4-phenoxyphenyl)-4H-benzo[4,5]imidazo[1,2-b]pyrazole-3-carboxamide
在0℃，向含有7-氨基-2-(4-苯氧基苯基)-4H-苯並[4,5]咪唑並[1,2-b]吡唑-3-甲醯胺(45mg,0.11mmol)的DCM(10mL)溶液中，加入三乙胺(36mg,0.35mmol)。20min內，逐滴 滴加丙烯醯氯(11mg,0.12mmol)的DCM(2mL)溶液。TLC和LCMS監測反應，直到大部分起始原料被消耗。混合物在水(50mL)和DCM(20mL)中分配，水相再用DCM(20mL)萃取。合併有機相，飽和食鹽水洗滌，Na2SO4乾燥，濃縮，用製備矽膠板(DCM/MeOH=20/1)純化，得到灰色固體7-丙烯醯胺基-2-(4-苯氧基苯基)-4H-苯並[4,5]咪唑並[1,2-b]吡唑-3-甲醯胺(4mg,7.8%)。1H NMR(400MHz,CD3OD-d4)δ 8.25(s,1H),7.61(d,J=8.4Hz,2H),7.44(d,J=8.8Hz,1H),7.39(d,J=8.8Hz,1H),7.32(d,J=7.6Hz,1H),7.30(d,J=7.6Hz,1H),7.08(t,J=7.6Hz,1H),7.05-6.90(m,4H),6.42-6.25(m,2H),5.69(dd,J=9.6,1.9Hz,1H).MS(ESI)m/e[M+1]+ 438。 To 7-amino-2-(4-phenoxyphenyl)-4H-benzo[4,5]imidazo[1,2-b]pyrazole-3-carboxamide (45 mg) at 0 °C Triethylamine (36 mg, 0.35 mmol) was added to a solution of EtOAc (EtOAc). A solution of propylene sulfonium chloride (11 mg, 0.12 mmol) in DCM (2 mL) was then evaporated. The reaction was monitored by TLC and LCMS until most of the starting material was consumed. The mixture was partitioned between EtOAc (EtOAc)EtOAc. The combined organic phases were washed with brine, dried Na 2 SO 4, concentrated and purified by preparative plate silica gel (DCM / MeOH = 20/1 ), to give a gray solid 7- acrylamide-2- (4-phenoxy Phenyl)-4H-benzo[4,5]imidazo[1,2-b]pyrazole-3-carboxamide (4 mg, 7.8%). 1 H NMR (400 MHz, CD 3 OD-d 4 ) δ 8.25 (s, 1H), 7.61 (d, J = 8.4 Hz, 2H), 7.44 (d, J = 8.8 Hz, 1H), 7.39 (d, J = 8.8 Hz, 1H), 7.32 (d, J = 7.6 Hz, 1H), 7.30 (d, J = 7.6 Hz, 1H), 7.08 (t, J = 7.6 Hz, 1H), 7.05-6.90 (m, 4H) ), 6.42 - 6.25 (m, 2H), 5.69 (dd, J = 9.6, 1.9 Hz, 1H). MS (ESI) m/e [M+1] + 438.
實施例24：化合物68-69的合成Example 24: Synthesis of Compound 68-69
化合物68：8-氨基-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]喹唑啉-3-甲醯胺 Compound 68 : 8-amino-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]quinazoline-3-carboxamide
步驟1：(2-氟-4-硝基苯基)甲醇 Step 1: (2-Fluoro-4-nitrophenyl)methanol
向含有2-氟-4-硝基苯甲醛(1.0g,5.92mmol)的CH3OH(10mL)溶液中，加入NaBH4(814mg,22mmol)。室 溫下攪拌15分鐘後，將混合物濃縮。殘餘物在EA(100mL)和飽和食鹽水(100mL)中分配，合併有機相，飽和食鹽水(100mL×2)洗滌，Na2SO4乾燥，濃縮得到紅色固體(2-氟-4-硝基苯基)甲醇(1.0g,99%)。MS(ESI)m/e[M+1]+ 172.0。 To CH 3 OH (10mL) solution containing 2-fluoro-4-nitrobenzaldehyde (1.0g, 5.92mmol) was added NaBH 4 (814mg, 22mmol). After stirring at room temperature for 15 minutes, the mixture was concentrated. The residue was partitioned between EA (100mL) and saturated brine (100 mL), washed, dried combined organic phases with saturated brine (100mL × 2) Na 2 SO 4, and concentrated to give a red solid (2-fluoro-4-nitro Phenyl)methanol (1.0 g, 99%). MS (ESI) m / e [M+1] + 172.0.
步驟2：2-(2-氟-4-硝基苄氧基)-四氫-2H-吡喃 Step 2: 2-(2-Fluoro-4-nitrobenzyloxy)-tetrahydro-2H-pyran
向(2-氟-4-硝基苯基)甲醇(755mg,4.42mmol)的DCM(10mL)溶液中，加入對TsOH(100mg,0.13mmol)和DHP(408mg,4.86mmol)。室溫攪拌16小時後，將混合物濃縮。殘餘物在EA(100mL)和飽和食鹽水(100mL)中分配。合併有機相，飽和食鹽水(100mL×2)洗滌，Na2SO4乾燥，濃縮，矽膠層析柱(PE/EA洗脫)純化，得到無色油狀物2-(2-氟-4-硝基苄氧基)-四氫-2H-吡喃(900mg,80%)。1H NMR(400MHz,DMSO-d6)δ 8.34(dd,J=3.0,6.2Hz,1H),8.26-8.30(m,1H),7.53(t,J=9.2Hz,1H),4.82-4.76(m,2H),4.62(d,J=12.0Hz,1H),3.80-3.74(m,1H),3.52-3.47(m,1H),1.76-1.64(m,2H),1.58-1.45(m,4H)。 To a solution of (2-fluoro-4-nitrophenyl)methanol (755 mg, 4.42 mmol) in EtOAc (EtOAc) After stirring at room temperature for 16 hours, the mixture was concentrated. The residue was partitioned between EA (100 mL) and brine. The organic phase was combined, washed with saturated brine (100 mL×2), dried over Na 2 SO 4 and concentrated and purified by silica gel chromatography (PE/EA elution) to give 2-(2-fluoro-4-nitrobenzene Benzyloxy)-tetrahydro-2H-pyran (900 mg, 80%). 1 H NMR (400MHz, DMSO- d 6) δ 8.34 (dd, J = 3.0,6.2Hz, 1H), 8.26-8.30 (m, 1H), 7.53 (t, J = 9.2Hz, 1H), 4.82-4.76 (m, 2H), 4.62 (d, J = 12.0 Hz, 1H), 3.80-3.74 (m, 1H), 3.52-3.47 (m, 1H), 1.76-1.64 (m, 2H), 1.58-1.45 (m , 4H).
步驟3：5-氨基-1-(5-硝基-2-((四氫-2H-吡喃-2-基氧基)甲基)苯基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺 Step 3: 5-Amino-1-(5-nitro-2-((tetrahydro-2H-pyran-2-yloxy)methyl)phenyl)-3-(4-phenoxyphenyl) )-1H-pyrazole-4-carboxamide
向5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺(27.6mg,0.1mmol)的DMF(3mL)和CH3CN(5mL)的溶液中，加入2-(2-氟-4-硝基苄氧基)-四氫-2H-吡喃(25.5mg,0.1mmol)和K2CO3(27.6mg,0.2mmol)。在氮氣保護下，80℃攪拌16小時後，混合物濃縮，PE/EA重結晶，得到黃色固體5-氨基-1-(5-硝基-2-((四氫-2H-吡喃-2-基氧基)甲基)苯基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺(40mg,80%)。MS(ESI)m/e[M+1]+ 512.2。 5-amino-3- (4-phenoxyphenyl) acyl lH-pyrazole-4-amine (27.6mg, 0.1mmol) in DMF (3mL) and CH 3 CN (5mL) solution, 2-(2-Fluoro-4-nitrobenzyloxy)-tetrahydro-2H-pyran (25.5 mg, 0.1 mmol) and K 2 CO 3 (27.6 mg, 0.2 mmol). After stirring at 80 ° C for 16 hours under a nitrogen atmosphere, the mixture was concentrated and then recrystallized from PE/EA to afford 5-amino-1-(5-nitro-2-((tetrahydro-2H-pyran-2-) Baseoxy)methyl)phenyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide (40 mg, 80%). MS (ESI) m / e [ M + 1] + 512.2.
步驟4：5-氨基-1-(2-(羥甲基)-5-硝基苯基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺 Step 4: 5-Amino-1-(2-(hydroxymethyl)-5-nitrophenyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide
向5-氨基-1-(5-硝基-2-((四氫-2H-吡喃-2-基氧基)甲基)苯基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺(690mg,1.3mmol)的CH3CN(10mL)溶液中，加入鹽酸(3mL)。室溫攪拌15min後，混合物濃縮，得到黃色固體5-氨基-1-(2-(羥甲基)-5-硝基苯基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺(550mg,95%)。 To 5-amino-1-(5-nitro-2-((tetrahydro-2H-pyran-2-yloxy)methyl)phenyl)-3-(4-phenoxyphenyl)- 1H- pyrazole-4-acyl-amine (690mg, 1.3mmol) in CH 3 CN solution (10mL), was added hydrochloric acid (3mL). After stirring at room temperature for 15 min, the mixture was concentrated to give a white solid, 5-amino-1-(2-(hydroxymethyl)-5-nitrophenyl)-3-(4-phenoxyphenyl)-1H-py Zolcon-4-carbamide (550 mg, 95%).
步驟5：5-氨基-1-(2-甲醯基-5-硝基苯基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺 Step 5: 5-Amino-1-(2-carbamimido-5-nitrophenyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide
向5-氨基-1-(2-(羥甲基)-5-硝基苯基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺(550mg,1.24mmol)的DCM(20mL)溶液中，加入MnO2(500mg,5.75mmol)。室溫攪拌16小時後，混合物過濾。濃縮濾液，得到的黃色固體5-氨基-1-(2-甲醯基-5-硝基苯基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺(400mg,73%)。 To 5-amino-1-(2-(hydroxymethyl)-5-nitrophenyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide (550 mg, 1.24) mmol) in DCM (20mL) was added MnO 2 (500mg, 5.75mmol). After stirring at room temperature for 16 hours, the mixture was filtered. The filtrate was concentrated to give a yellow solid, 5-amino-1-(2-carbamido-5-nitrophenyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide (400 mg, 73%).
步驟6：8-硝基-2-(4-苯氧基苯基)唑並[1,5-a]喹唑啉-3-甲醯胺 Step 6: 8-Nitro-2-(4-phenoxyphenyl)oxazolo[1,5-a]quinazoline-3-carboxamide
向5-氨基-1-(2-甲醯基-5-硝基苯基)-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺(400mg,0.9mmol)的CH3OH(5mL)和DCM(5mL)溶液中，加入醋酸(1滴)。室溫攪拌16小時後，混合物濃縮，矽膠層析柱(PE/EA洗脫)純化，得到黃色固體8-硝基-2-(4-苯氧基苯基)唑並[1,5-a]喹唑啉-3-甲醯胺(240mg,63%)。MS(ESI)m/e[M+1]+ 425.8。 To 5-amino-1-(2-carbamimido-5-nitrophenyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide (400 mg, 0.9 mmol) in CH 3 OH (5mL) and DCM (5mL) was added acetic acid (1 drop). After stirring at room temperature for 16 hours, the mixture was concentrated and purified on a silica gel column (PE/EA elution) to give a yellow solid, 8-nitro-2-(4-phenoxyphenyl) azole [1,5-a Uquinazoline-3-carboxamide (240 mg, 63%). MS (ESI) m / e [M+1] + 425.8.
步驟7：8-硝基-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]喹唑啉-3-甲醯胺 Step 7: 8-Nitro-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]quinazoline-3-carboxamide
室溫下，向8-硝基-2-(4-苯氧基苯基)唑並[1,5-a]喹唑啉-3-甲醯胺(240mg,0.57mmol)的EtOH(10mL)和DCM(10mL)溶液中，加入NaBH4(86mg,2.26mmol)。室溫攪拌20分鐘後，加入水(10mL)。混合物濃縮，加入水(5mL)並過濾。濾餅用甲基叔丁基醚(30mL)洗滌，乾燥得到黃色固體8-硝基-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]喹唑啉-3-甲醯胺(200mg,83%)。MS(ESI)m/e[M+1]+ 427.9。 To 8-nitro-2-(4-phenoxyphenyl)oxazolo[1,5-a]quinazolin-3-carboxamide (240 mg, 0.57 mmol) in EtOH (10 mL) and DCM (10mL) was added NaBH 4 (86mg, 2.26mmol). After stirring at room temperature for 20 minutes, water (10 mL) was added. The mixture was concentrated, water (5 mL) was added and filtered. The filter cake was washed with methyl tert-butyl ether (30 mL) and dried to give a yellow solid, 8-nitro-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a Uquinazoline-3-carboxamide (200 mg, 83%). MS (ESI) m / e [M+1] + 427.9.
步驟8：8-氨基-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]喹唑啉-3-甲醯胺和8-氨基-2-(4-苯氧基苯基)吡唑並[1,5-a]喹唑啉-3-甲醯胺 Step 8: 8-Amino-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]quinazoline-3-carboxamide and 8-amino-2 -(4-phenoxyphenyl)pyrazolo[1,5-a]quinazoline-3-carboxamide
向8-硝基-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]喹唑啉-3-甲醯胺(200mg,0.47mmol)的CH3OH(30mL)和DCM(30mL)溶液中，加10% w/w Pd/C(100mg)。室溫攪拌1小時後，將混合物過濾。濃縮濾液，得到黃色固體粗品8-氨基-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]喹唑啉-3-甲醯胺和8-氨基-2-(4- 苯氧基苯基)吡唑並[1,5-a]喹唑啉-3-甲醯胺(130mg,70%)。MS(ESI)m/e[M+1]+ 398.0,395.9。 To 8-nitro-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]quinazoline-3-carboxamide (200 mg, 0.47 mmol) CH 3 OH (30mL) and DCM (30mL) solution was added 10% w / w Pd / C (100mg). After stirring at room temperature for 1 hour, the mixture was filtered. The filtrate was concentrated to give a crude yellow solid, 8-amino-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]quinazoline-3-carboxamide and 8 -Amino-2-(4-phenoxyphenyl)pyrazolo[1,5-a]quinazolin-3-carboxamide (130 mg, 70%). MS (ESI) m / e [ M + 1] + 398.0,395.9.
步驟9：8-氨基-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]喹唑啉-3-甲醯胺 Step 9: 8-Amino-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]quinazoline-3-carboxamide
向8-氨基-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]喹唑啉-3-甲醯胺和8-氨基-2-(4-苯氧基苯基)吡唑並[1,5-a]喹唑啉-3-甲醯胺(130mg,0.33mmol)的DCM(10mL)和CH3OH(10mL)溶液中，加入NaBH4(277mg,3.3mmol)。室溫攪拌15min後，加入水(50mL)。濃縮混合物，並過濾。濾餅用水(50mL×2)洗滌，得到黃色固體8-氨基-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]喹唑啉-3-甲醯胺(60mg,46%)。1H NMR(400MHz,DMSO-d6)δ 7.60(d,J=8.0Hz,2H),7.44(d,J=7.6Hz,1H),7.42(d,J=7.6Hz,1H),7.28(d,J=7.6Hz,1H),7.18(t,J=7.6Hz,1H),7.14-7.16(m,4H),6.81(s,1H),6.52(d,J=8.0Hz,1H),6.43(s,1H),5.16(s,2H),4.37(s,2H).MS(ESI)m/e[M+1]+ 397.9。 To 8-amino-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]quinazoline-3-carboxamide and 8-amino-2-( 4-phenoxyphenyl) pyrazolo [1,5-a] quinazoline-3-acyl-amine (130mg, 0.33mmol) in DCM (10mL) and CH 3 OH (10mL) was added NaBH 4 (277 mg, 3.3 mmol). After stirring at room temperature for 15 min, water (50 mL) was added. The mixture was concentrated and filtered. The filter cake was washed with water (50 mL×2) to give a yellow solid, 8-amino-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]quinazoline-3 - Formamide (60 mg, 46%). 1 H NMR (400MHz, DMSO- d 6) δ 7.60 (d, J = 8.0Hz, 2H), 7.44 (d, J = 7.6Hz, 1H), 7.42 (d, J = 7.6Hz, 1H), 7.28 ( d, J = 7.6 Hz, 1H), 7.18 (t, J = 7.6 Hz, 1H), 7.14 - 7.16 (m, 4H), 6.81 (s, 1H), 6.52 (d, J = 8.0 Hz, 1H), 6.43 (s, 1H), 5.16 (s, 2H), 4.37 (s, 2H). MS (ESI) m/e [M+1] + 397.9.
化合物69：8-丙烯醯胺基-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]喹唑啉-3-甲醯胺 Compound 69 : 8-propenylamino-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]quinazoline-3-carboxamide
目標產品由化合物68和丙烯醯氯，根據化合物8的類似方法合成。1H NMR(400MHz,DMSO-d6)δ 10.26(s,1H),7.39-7.63(m,4H),7.52(d,J=8.8Hz,1H),7.45(d,J=8.0Hz,1H),7.42(d,J=8.0Hz,1H),7.19(t,J=8.0Hz,1H),7.14-7.07(m,4H),7.01(s,1H),6.44(dd,J=10.4,17.0Hz,1H),6.26(dd,J=1.6,17.0Hz,1H),5.77(dd,J=1.6,10.4Hz,1H),4.51(s,2H).MS(ESI)m/e[M+1]+ 451.9。 The target product was synthesized from Compound 68 and propylene chloride in a similar manner to Compound 8 . 1 H NMR (400MHz, DMSO- d 6) δ 10.26 (s, 1H), 7.39-7.63 (m, 4H), 7.52 (d, J = 8.8Hz, 1H), 7.45 (d, J = 8.0Hz, 1H ), 7.42 (d, J = 8.0 Hz, 1H), 7.19 (t, J = 8.0 Hz, 1H), 7.14 - 7.07 (m, 4H), 7.01 (s, 1H), 6.44 (dd, J = 10.4, 17.0 Hz, 1H), 6.26 (dd, J = 1.6, 17.0 Hz, 1H), 5.77 (dd, J = 1.6, 10.4 Hz, 1H), 4.51 (s, 2H). MS (ESI) m/e [M +1] + 451.9.
實施例25：化合物70-72的合成Example 25: Synthesis of Compound 70-72
化合物70：8-硝基-2-(4-苯氧基苯基)-5,6-二氫-4H-苯並[f]吡唑並[1,5-a][1,3]二氮雜-3-甲醯胺 Compound 70 : 8-nitro-2-(4-phenoxyphenyl)-5,6-dihydro-4H-benzo[f]pyrazolo[1,5-a][1,3] Aza-3-carboxamide
步驟1：2-(2-氟-5-硝基苯基)乙醇 Step 1: 2-(2-Fluoro-5-nitrophenyl)ethanol
室溫下將2-(2-氟-5-硝基苯基)乙酸(2.0g,10mmol)溶入四氫呋喃(50mL)溶液中，向其中滴加硼烷二甲硫醚絡合物(4.0g,25mmol)，滴加完畢將溫度升到60℃，攪拌12小時。冷卻到 室溫，向反應液中慢慢加入甲醇(20mL)，真空旋幹。殘餘物經柱層析(200-300目，石油醚：乙酸乙酯=2：1)，得到無色油狀液體(1.6g，86.1%)。1H NMR(DMSO-d 6)δ 8.27(dd,J=3.2,6.4Hz,1 H),8.19-8.14(m,1 H),8.45(t,J=9.2Hz,1 H),4.80(t,J=5.6Hz,1 H),3.66(dt,J=5.6,6.2Hz,1 H),2.86(t,J=6.2Hz,2 H).MS(ESI)m/e[M+1]+ 186。 2-(2-Fluoro-5-nitrophenyl)acetic acid (2.0 g, 10 mmol) was dissolved in tetrahydrofuran (50 mL) at room temperature, and borane dimethyl sulfide complex (4.0 g) was added dropwise thereto. , 25 mmol), the temperature was raised to 60 ° C after the dropwise addition, and stirred for 12 hours. After cooling to room temperature, methanol (20 mL) was slowly added to the reaction mixture, which was evaporated to dryness. The residue was subjected to EtOAc EtOAcjjjjjjjj 1 H NMR (DMSO- d 6 ) δ 8.27 (dd, J = 3.2, 6.4 Hz, 1 H), 8.19-8.14 (m, 1 H), 8.45 (t, J = 9.2 Hz, 1 H), 4.80 ( t, J = 5.6 Hz, 1 H), 3.66 (dt, J = 5.6, 6.2 Hz, 1 H), 2.86 (t, J = 6.2 Hz, 2 H). MS (ESI) m/e [M+1 ] + 186.
步驟2：8-硝基-2-(4-苯氧基苯基)-5,6-二氫-4H-苯並[f]吡唑並[1,5-a][1,3]二氮雜-3-甲醯胺 Step 2: 8-Nitro-2-(4-phenoxyphenyl)-5,6-dihydro-4H-benzo[f]pyrazolo[1,5-a][1,3] Aza-3-carboxamide
將5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺(30mg,0.10mmol)溶於DMF(5.0mL)，向其中加入碳酸鉀(28mg,0.20mmol)和2-(2-氟-5-硝基苯基)乙醇(37mg,0.20mmol)。混合物升溫到80℃，攪拌16小時。冷卻至室溫，減壓旋蒸，向剩餘物中加入乙酸乙酯(15mL)和水(15mL)，水相用乙酸乙酯萃取(10mL×3)，合併有機相，飽和食鹽水(10mL)洗滌，無水硫酸鈉乾燥，過濾，濃縮，製備矽膠板純化(DCM/CH3OH=20/1)得到產品(5.0mg,11.1%)。1H NMR(DMSO-d 6)δ 8.35(dd,J=3.2,4.0Hz,1 H),8.27(d,J=2.6Hz,1 H),8.22(dd,J=2.6,9.2Hz,1 H),8.12(d,J=9.2Hz,1 H),7.64-7.60(m,2 H),7.45-7.41(m,2 H),7.22-7.17(m,1 H),7.14-7.09(m,4 H), 3.72-3.65(m,2 H),3.29-3.24(m,2 H).MS(ESI)m/e[M+1]+ 442。 5-Amino-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide (30 mg, 0.10 mmol) was dissolved in DMF (5.0 mL) and potassium carbonate (28 mg, 0.20) Methyl) and 2-(2-fluoro-5-nitrophenyl)ethanol (37 mg, 0.20 mmol). The mixture was warmed to 80 ° C and stirred for 16 hours. The mixture was cooled to room temperature, and the mixture was evaporated to dryness. EtOAcjjjjjjjjjjjjjjjjjj , dried over anhydrous sodium sulfate, filtered, concentrated and purified by preparative plate silica gel (DCM / CH 3 OH = 20 /1) to give the product (5.0mg, 11.1%). 1 H NMR (DMSO- d 6 ) δ 8.35 (dd, J = 3.2, 4.0 Hz, 1 H), 8.27 (d, J = 2.6 Hz, 1 H), 8.22 (dd, J = 2.6, 9.2 Hz, 1 H), 8.12 (d, J = 9.2 Hz, 1 H), 7.64-7.60 (m, 2 H), 7.45-7.41 (m, 2 H), 7.22-7.17 (m, 1 H), 7.14-7.09 ( m, 4 H), 3.72-3.65 (m, 2 H), 3.29-3.24 (m, 2 H). MS (ESI) m/e [M+1] + 442.
化合物71：8-氨基-2-(4-苯氧基苯基)-5,6-二氫-4H-苯並[f]吡唑並[1,5-a][1,3]二氮雜-3-甲醯胺 Compound 71: 8-amino-2-(4-phenoxyphenyl)-5,6-dihydro-4H-benzo[f]pyrazolo[1,5-a][1,3]diazepine Hetero-3-carboxamide
將8-硝基-2-(4-苯氧基苯基)-5,6-二氫-4H-苯並[f]吡唑並[1,5-a][1,3]二氮雜-3-甲醯胺(80mg,0.18mmol)溶入乙醇(20mL)，加入10% w/w Pd/C(20mg)，氫氣條件下室溫攪拌3小時。過濾，濾餅用甲醇(20mL)洗滌，濾液濃縮，殘餘物用製備板純化(DCM/CH3OH=20/1)，得到白色固體(20mg,26.8%)。1H NMR(DMSO-d 6)δ 7.73-7.69(m,1 H),7.59-7.55(m,2 H),7.45-7.40(m,3 H),7.21-7.16(m,1 H),7.13-7.07(m,4 H),6.59-6.54(m,1 H),6.50-6.47(m,1 H),5.70-5.40(brs,2 H),3.64-3.59(m,2 H),2.99-2.94(m,2 H).MS(ESI)m/e[M+1]+ 412。 8-Nitro-2-(4-phenoxyphenyl)-5,6-dihydro-4H-benzo[f]pyrazolo[1,5-a][1,3]diazepine 3-Methylguanamine (80 mg, 0.18 mmol) was dissolved in ethanol (20 mL), and 10% w/w Pd/C (20 mg) was added, and the mixture was stirred at room temperature for 3 hours under hydrogen. The filter cake washed with methanol (20mL), and the filtrate was concentrated, the residue was purified by preparative TLC (DCM / CH 3 OH = 20 /1) to give a white solid (20mg, 26.8%). 1 H NMR (DMSO- d 6 ) δ 7.73-7.69 (m, 1 H), 7.59-7.55 (m, 2 H), 7.45-7.40 (m, 3 H), 7.21-7.16 (m, 1 H), 7.13-7.07 (m, 4 H), 6.59-6.54 (m, 1 H), 6.50-6.47 (m, 1 H), 5.70-5.40 (brs, 2 H), 3.64-3.59 (m, 2 H), 2.99-2.94 (m, 2 H). MS (ESI) m/e [M+1] + 412.
化合物72：8-丙烯醯胺基-2-(4-苯氧基苯基)-5,6-二氫-4H-苯並[f]吡唑並[1,5-a][1,3]二氮雜-3-甲醯胺 Compound 72: 8-Acrylamino-2-(4-phenoxyphenyl)-5,6-dihydro-4H-benzo[f]pyrazolo[1,5-a][1,3 Diaza-3-carboxamide
目標產物由化合物71和丙烯醯氯，參照化合物8的類似方法合成。1H NMR(DMSO-d 6)δ 10.25(s,1 H),7.95(t,J=4.0Hz,1 H),7.76(d,J=8.8Hz,1 H),7.65-7.58(m,4 H),7.46-7.41(m,2 H),7.21-7.17(m,1 H),7.13-7.08(m,4 H),6.44(dd,J=10.0,16.8Hz,1 H),6.27(dd,J=2.0,16.8Hz,1 H),5.77(dd,J=2.0,10.0Hz,1 H),3.68-3.65(m,2 H),3.05-3.03(m,2 H).MS(ESI)m/e[M+1]+ 466。 The target product was synthesized from Compound 71 and propylene sulfonium chloride in a similar manner to Compound 8 . 1 H NMR (DMSO- d 6 ) δ 10.25 (s, 1 H), 7.95 (t, J = 4.0 Hz, 1 H), 7.76 (d, J = 8.8 Hz, 1 H), 7.65-7.58 (m, 4 H), 7.46-7.41 (m, 2 H), 7.21-7.17 (m, 1 H), 7.13-7.08 (m, 4 H), 6.44 (dd, J = 10.0, 16.8 Hz, 1 H), 6.27 (dd, J = 2.0, 16.8 Hz, 1 H), 5.77 (dd, J = 2.0, 10.0 Hz, 1 H), 3.68-3.65 (m, 2 H), 3.05-3.03 (m, 2 H).MS (ESI) m/e [M+1] + 466.
實施例26：化合物73-75的合成Example 26: Synthesis of Compound 73-75
化合物73：8-硝基-5-氧代-2-(4-苯氧基苯基)-5,6-二氫-4H-苯並[f]吡唑並[1,5-a][1,3]二氮雜-3-甲醯胺 Compound 73 : 8-nitro-5-oxo-2-(4-phenoxyphenyl)-5,6-dihydro-4H-benzo[f]pyrazolo[1,5-a][ 1,3]diaza-3-carboxamide
步驟1：2-(2-氟-5-硝基苯基)乙酸甲酯 Step 1: Methyl 2-(2-fluoro-5-nitrophenyl)acetate
將2-(2-氟-5-硝基苯基)乙酸(1.0g,5.0mmol)溶入甲醇中(20mL)，向其中逐滴滴加濃硫酸(0.50mL)，滴加完畢，將反應溫度升至80℃反應3小時。冷卻至室溫，將反應倒入水(20 mL)中，濃縮蒸出甲醇。水相用乙酸乙酯萃取(20mL×2)，合併有機相，用飽和食鹽水(10mL)洗滌一次，無水硫酸鈉乾燥，過濾，濃縮，得到無色油狀物(1.0g，93.4%)。MS(ESI)m/e[M+1]+ 214。 2-(2-Fluoro-5-nitrophenyl)acetic acid (1.0 g, 5.0 mmol) was dissolved in methanol (20 mL), and concentrated sulfuric acid (0.50 mL) was added dropwise thereto, and the reaction was completed. The temperature was raised to 80 ° C for 3 hours. After cooling to room temperature, the reaction was poured into water (20 mL). The aqueous layer was extracted with EtOAc (EtOAc)EtOAc. MS (ESI) m/e [M+1] + 214.
步驟2：8-硝基-5-氧代-2-(4-苯氧基苯基)-5,6-二氫-4H-苯並[f]吡唑並[1,5-a][1,3]二氮雜-3-甲醯胺 Step 2: 8-Nitro-5-oxo-2-(4-phenoxyphenyl)-5,6-dihydro-4H-benzo[f]pyrazolo[1,5-a][ 1,3]diaza-3-carboxamide
將5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺(30mg,0.10mmol)溶入DMF(5mL)，向其中加入碳酸鉀(28mg,0.20mmol)和甲基2-(2-氟-5-硝基苯基)乙酸甲酯(21mg,0.10mmol)，將混合物溫度升至80℃，攪拌反應16小時。冷卻至室溫，濃縮，蒸去DMF，剩餘物加入二氯甲烷(10mL)和水(10mL)，水相用二氯甲烷萃取(10mL×2)，合併有機相，用飽和氯化鈉(10mL)洗滌，無水硫酸鈉乾燥，過濾，濃縮，用製備板純化(DCM/CH3OH=20/1)，得到產品(10mg,21.9%)。1H NMR(DMSO-d 6)δ 10.66(br s,1 H),8.51(d,J=2.6Hz,1 H),8.37(dd,J=2.6,9.2Hz,1 H),8.07(d,J=9.2Hz,1 H),7.85(d,J=8.8Hz,2 H),7.65-7.53(br s,2 H),7.45-7.40(m,2 H),7.22-7.16(m,1 H),7.12-7.05(m,4 H),3.92(s,2 H).MS(ESI)m/e[M+1]+ 456。 5-Amino-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide (30 mg, 0.10 mmol) was dissolved in DMF (5 mL) and potassium carbonate (28 mg, 0.20 mmol) And methyl 2-(2-fluoro-5-nitrophenyl)acetic acid methyl ester (21 mg, 0.10 mmol), the mixture was warmed to 80 ° C, and the reaction was stirred for 16 hours. After cooling to room temperature, concentrating, DMF was evaporated, m~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ ), dried over anhydrous sodium sulfate, filtered, concentrated and purified by preparative TLC (DCM / CH by 3 OH = 20/1), to give the product (10mg, 21.9%). 1 H NMR (DMSO- d 6 ) δ 10.66 (br s, 1 H), 8.51 (d, J = 2.6 Hz, 1 H), 8.37 (dd, J = 2.6, 9.2 Hz, 1 H), 8.07 (d) , J = 9.2 Hz, 1 H), 7.85 (d, J = 8.8 Hz, 2 H), 7.65-7.53 (br s, 2 H), 7.45-7.40 (m, 2 H), 7.22 - 7.16 (m, 1 H), 7.12-7.05 (m, 4 H), 3.92 (s, 2 H). MS (ESI) m/e [M+1] + 456.
化合物74：8-氨基-5-氧代-2-(4-苯氧基苯基)-5,6-二氫-4H-苯並[f]吡唑並[1,5-a][1,3]二氮雜-3-甲醯胺 Compound 74 : 8-amino-5-oxo-2-(4-phenoxyphenyl)-5,6-dihydro-4H-benzo[f]pyrazolo[1,5-a][1 ,3]diazepine-3-carboxamide
化合物74由8-硝基-5-氧代-2-(4-苯氧基苯基)-5,6-二氫-4H-苯並[f]吡唑並[1,5-a][1,3]二氮雜-3-甲醯胺，參照化合物71的類似方法合成。1H NMR(CD3OD-d 4)δ 7.72-7.67(m,2 H),7.52(d,J=8.8Hz,1 H),7.42-7.37(m,2 H),7.19-7.14(m,1 H),7.11-7.05(m,4 H),6.80(dd,J=2.6,8.8Hz,1 H),6.69(d,J=2.6Hz,1 H),3.59(s,2 H).MS(ESI)m/e[M+1]+ 426.1。 Compound 74 consists of 8-nitro-5-oxo-2-(4-phenoxyphenyl)-5,6-dihydro-4H-benzo[f]pyrazolo[1,5-a][ 1,3]diazepine-3-carboxamide was synthesized in a similar manner to the compound 71 . 1 H NMR (CD 3 OD- d 4 ) δ 7.72-7.67 (m, 2 H), 7.52 (d, J = 8.8 Hz, 1 H), 7.42-7.37 (m, 2 H), 7.19-7.14 (m) , 1 H), 7.11 - 7.05 (m, 4 H), 6.80 (dd, J = 2.6, 8.8 Hz, 1 H), 6.69 (d, J = 2.6 Hz, 1 H), 3.59 (s, 2 H) MS (ESI) m / e [M + 1] + 426.1.
化合物75：8-丙烯醯胺基-5-氧代-2-(4-苯氧基苯基)-5,6-二氫-4H-苯並[f]吡唑並[1,5-a][1,3]二氮雜-3-甲醯胺 Compound 75 : 8-propenylamino-5-oxo-2-(4-phenoxyphenyl)-5,6-dihydro-4H-benzo[f]pyrazolo[1,5-a ][1,3]diazepine-3-carboxamide
化合物75由8-氨基-5-氧代-2-(4-苯氧基苯基)-5,6-二氫-4H-苯並[f]吡唑並[1,5-α][1,3]二氮雜-3-甲醯胺和丙烯醯氯，參照化合物8的類似方法合成。1H NMR(CD3OD-d 4)δ 7.85-7.79(m,3 H),7.75-7.71(m,2 H),7.43-7.37(m,2 H),7.20-7.14(m,1 H),7.12-7.06(m,4 H),6.50-6.35(m,2 H),5.81(dd,J=2.6,9.0Hz,1 H),3.75(s,2 H).MS(ESI)m/e[M+1]+ 480.1。 Compound 75 consists of 8-amino-5-oxo-2-(4-phenoxyphenyl)-5,6-dihydro-4H-benzo[f]pyrazolo[1,5- α ][1 , 3] diaza-3-carboxamide and acrylonitrile chloride, synthesized in a similar manner to that of compound 8 . 1 H NMR (CD 3 OD- d 4 ) δ 7.85-7.79 (m, 3 H), 7.75-7.71 (m, 2 H), 7.43-7.37 (m, 2 H), 7.20-7.14 (m, 1 H ), 7.12-7.06 (m, 4 H), 6.50-6.35 (m, 2 H), 5.81 (dd, J = 2.6, 9.0 Hz, 1 H), 3.75 (s, 2 H). MS (ESI) m /e[M+1] + 480.1.
實施例27：化合物76-79的合成Example 27: Synthesis of Compound 76-79
化合物76：7-硝基-5-氧代-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]喹唑啉-3-甲醯胺 Compound 76 : 7-nitro-5-oxo-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]quinazoline-3-carboxamide
氮氣保護下，5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺(220mg，0.75mmol)，2-氯-5-硝基苯甲酸甲酯(160mg,0.75mmol)和K2CO3(155mg,1.13mmol)在DMF(10mL)中的混合物，80℃加熱16小時。將反應物倒入水(30ml)中，乙酸乙酯(20mL×3)萃取。合併有機相，Na2SO4乾燥，濃縮，殘餘物矽膠層析柱(在PE中，EA從10%梯度到50%)純化，得到黃色固體7-硝基-5-氧代-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]喹唑啉-3-甲醯胺(85mg,27.3%)。MS(ESI,m/e)[M+1]+ 442.1。 5-Amino-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide (220 mg, 0.75 mmol), methyl 2-chloro-5-nitrobenzoate (under nitrogen) mixture in DMF (10mL) is 160mg, 0.75mmol) and K 2 CO 3 (155mg, 1.13mmol ), 80 ℃ was heated for 16 h. The reaction was poured into water (30 mL) andEtOAcEtOAc The organic phases were combined, Na dried 2 SO 4, concentrated and the residue was silica gel column chromatography (in the PE, EA gradient from 10% to 50%) to give a yellow solid 7-nitro-5-oxo-2- ( 4-Phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]quinazoline-3-carboxamide (85 mg, 27.3%). MS (ESI, m/e) [M+1] + 4421.
化合物77：7-氨基-5-氧代-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]喹唑啉-3-甲醯胺 Compound 77 : 7-Amino-5-oxo-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]quinazoline-3-carboxamide
化合物77是由7-硝基-5-氧代-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]喹唑啉-3-甲醯胺，根據化合物71的類似方法合成。1H NMR(400MHz,DMSO-d6)δ 10.88(br s,1H),7.80(d,J=8.4Hz,1H),7.71(d,J=6.8Hz,2H),7.36-7.42(m,2H),7.27(s,1H),7.02-7.17(m,6H),5.62(s,2H).MS(ESI,m/e)[M+1]+ 412.1。 Compound 77 is from 7-nitro-5-oxo-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]quinazoline-3-carboxamidine The amine was synthesized according to a similar method of Compound 71 . 1 H NMR (400MHz, DMSO- d 6) δ 10.88 (br s, 1H), 7.80 (d, J = 8.4Hz, 1H), 7.71 (d, J = 6.8Hz, 2H), 7.36-7.42 (m, 2H), 7.27 (s, 1H), 7.02-7.17 (m, 6H), 5.62 (s, 2H). MS (ESI, m/e) [M+1] + 412.1.
化合物78：7-丙烯醯氨基-5-氧代-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]喹唑啉-3-甲醯胺 Compound 78 : 7-Acrylamino-5-oxo-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]quinazoline-3-carboxamidine amine
化合物78是由7-氨基-5-氧代-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]喹唑啉-3-甲醯胺和丙烯醯氯，根據化合物8的類似方法合成。1H NMR(400MHz,DMSO-d6)δ 11.37(s,1H),10.77(s,1H),8.65(s,1H),8.24(d,J=8Hz,1H),8.15(d,J=8Hz,1H),7.85(d,J=7.6Hz,2H),7.48-7.52(m,2H),7.2-7.15(m,5H),6.57(dd,J=9.2,18.0Hz,1H),6.37(d,J=18.0Hz,1H),5.87(d,J=9.2Hz,1H).MS(ESI,m/e)[M+1]+ 466.1。 Compound 78 is from 7-amino-5-oxo-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]quinazoline-3-carboxamide And propylene chloride, synthesized in a similar manner to compound 8 . 1 H NMR (400MHz, DMSO- d 6) δ 11.37 (s, 1H), 10.77 (s, 1H), 8.65 (s, 1H), 8.24 (d, J = 8Hz, 1H), 8.15 (d, J = 8 Hz, 1H), 7.85 (d, J = 7.6 Hz, 2H), 7.48-7.52 (m, 2H), 7.2-7.15 (m, 5H), 6.57 (dd, J = 9.2, 18.0 Hz, 1H), 6.37 (d, J = 18.0 Hz, 1H), 5.87 (d, J = 9.2 Hz, 1H). MS (ESI, m/e) [M+1] + 466.1.
化合物79：8-氨基-5-氧代-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]喹唑啉-3-甲醯胺 Compound 79 : 8-amino-5-oxo-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]quinazoline-3-carboxamide
目標產品由2-氯-4-硝基苯甲酸甲酯和5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺，參照化合物76和77的類似方法合成。1H NMR(400MHz,DMSO-d6)δ 10.61(s,1H),7.77(d,J=8.4Hz,1H),7.71(d,J=8.0Hz,2H),7.44-7.36(m,2H),7.19-7.02(m,6H),6.64(d,J=8.4Hz,1H),6.55(br s,2H).MS(ESI,m/e)[M+1]+ 412.1。 The target product consists of methyl 2-chloro-4-nitrobenzoate and 5-amino-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide, analogous to compounds 76 and 77 . Method synthesis. 1 H NMR (400MHz, DMSO- d 6) δ 10.61 (s, 1H), 7.77 (d, J = 8.4Hz, 1H), 7.71 (d, J = 8.0Hz, 2H), 7.44-7.36 (m, 2H ), 7.19-7.02 (m, 6H), 6.64 (d, J = 8.4 Hz, 1H), 6.55 (br s, 2H). MS (ESI, m/e) [M+1] + 412.1.
實施例28：化合物80-81的合成Example 28: Synthesis of Compound 80-81
化合物80：5-氧代-2-(4-苯氧基苯基)-7-(4-哌啶基)-4,5-二氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 80 : 5-oxo-2-(4-phenoxyphenyl)-7-(4-piperidinyl)-4,5-dihydropyrazolo[1,5-a]pyrimidine-3- Formamide
步驟1：4-(2,2-二甲基-4,6-二氧代-1,3-二氧六環-5-羰基)-哌啶-1-甲酸叔丁酯 Step 1: 4-(2,2-Dimethyl-4,6-dioxo-1,3-dioxocyclo-5-carbonyl)-piperidine-1-carboxylic acid tert-butyl ester
向1-(甲酸叔丁酯)哌啶-4-羧酸(1.15g,5mmol)和DMAP(61mg,0.5mmol)在DCM(50mL)中的混合物中，加入DCC(1.14g,5.5mmol)和2,2-甲基-1,3-二氧六環-4,6-二酮 (0.8g,5.5mmol)。反應混合物室溫攪拌16個小時，過濾。濾液濃縮，得到黃色油狀粗品4-(2,2-二甲基-4,6-二氧代-1,3-二氧六環-5-羰基)-哌啶-1-甲酸叔丁酯(2g,crude)，未經純化直接用於下一步反應。MS(ESI)m/e[M+23]+ 378.1。 To a mixture of 1-(carboxylic acid tert-butyl ester) piperidine-4-carboxylic acid (1.15 g, 5 mmol) and EtOAc (EtOAc (EtOAc) 2,2-Methyl-1,3-dioxane-4,6-dione (0.8 g, 5.5 mmol). The reaction mixture was stirred at room temperature for 16 hours and filtered. The filtrate was concentrated to give 4-(2,2-dimethyl-4,6-dioxo-1,3-dioxocyclo-5-carbonyl)-piperidine-1-carboxylic acid tert-butyl ester as a yellow oil. (2g, crude), used directly in the next reaction without purification. MS (ESI) m / e [M+23] + 378.1.
步驟2：4-(3-乙氧基-3-氧代丙醯基)哌啶-1-甲酸叔丁酯 Step 2: 4-(3-Ethoxy-3-oxopropionyl)piperidine-1-carboxylic acid tert-butyl ester
4-(2,2-二甲基-4,6-二氧代-1,3-二氧六環-5-羰基)-哌啶-1-甲酸叔丁酯(2g,5.63mmol)的乙醇(50ml)溶液，加熱回流20個小時。減壓除去溶劑，殘餘物矽膠層析柱分離純化，DCM洗脫得到微紅色液體4-(3-乙氧基-3-氧代丙醯基)哌啶-1-甲酸叔丁酯(0.5g,30%)。MS(ESI)m/e[M+23]+ 322.2。 4-(2,2-Dimethyl-4,6-dioxo-1,3-dioxocyclo-5-carbonyl)-piperidine-1-carboxylic acid tert-butyl ester (2 g, 5.63 mmol) in ethanol (50 ml) solution, heated to reflux for 20 hours. The solvent was removed under reduced pressure. , 30%). MS (ESI) m / e [M+23] + 322.2.
步驟3：5-氧代-2-(4-苯氧基苯基)-7-(哌啶-4-基)-4,5-二氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 3: 5-Oxo-2-(4-phenoxyphenyl)-7-(piperidin-4-yl)-4,5-dihydropyrazolo[1,5-a]pyrimidine-3 -Procarbamide
5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲醯胺(412mg,1.4mmol)和4-(3-乙氧基-3-氧代丙醯基)哌啶-1-甲酸叔丁酯(420mg,1.4mmol)在HOAc(20mL)中的混合物，90℃攪拌16個小時。減壓除去溶劑，殘餘物在aq.NaHCO3和乙酸乙酯中分配。有機相飽和食鹽水洗滌，Na2SO4乾燥，減壓濃縮。殘餘物製 備HPLC純化，水相含0.1% TFA，25%到90% CH3CN梯度洗脫，收集合併目標產品，過夜凍幹，得到白色固體5-氧代-2-(4-苯氧基苯基)-7-(哌啶-4-基)-4,5-二氫吡唑並[1,5-a]嘧啶-3-甲醯胺(0.3g,50%)。1H NMR(400MHz,DMSO-d6)δ 11.71(br s,1H),8.68-8.65(m,1H),8.42-8.39(m,1H),7.74(d,J=8.4Hz,2H),7.47-7.41(m,2H),7.22-7.18(m,1H),7.14-7.09(m,4H),5.72(s,1H),3.50-3.35(m,2H),3.17-3.06(m,1H),3.01-2.87(m,2H),2.15-2.05(m,2H),1.83-1.72(m,2H).MS(ESI)m/e[M+1]+ 430.1。 5-amino-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide (412 mg, 1.4 mmol) and 4-(3-ethoxy-3-oxopropenyl) A mixture of piperidine-1-carboxylic acid tert-butyl ester (420 mg, 1.4 mmol) in EtOAc (20 mL) The solvent was removed, and the residue was partitioned between ethyl acetate and aq.NaHCO 3 under reduced pressure. The organic layer was washed with brine, dried over Na 2 CH 4 The residue was purified by preparative HPLC, an aqueous phase containing 0.1% TFA, 25% to 90% CH 3 CN gradient, to collect the target product were combined, lyophilized overnight, to give a white solid 5-oxo-2- (4-phenoxy Phenyl)-7-(piperidin-4-yl)-4,5-dihydropyrazolo[1,5-a]pyrimidine-3-carboxamide (0.3 g, 50%). 1 H NMR (400MHz, DMSO- d 6) δ 11.71 (br s, 1H), 8.68-8.65 (m, 1H), 8.42-8.39 (m, 1H), 7.74 (d, J = 8.4Hz, 2H), 7.47-7.41(m,2H),7.22-7.18(m,1H),7.14-7.09(m,4H),5.72(s,1H),3.50-3.35(m,2H),3.17-3.06(m,1H ), 3.01-2.87 (m, 2H), 2.15-2.05 (m, 2H), 1.83-1.72 (m, 2H). MS (ESI) m/e [M+1] + 430.1.
化合物81：5-氧代-2-(4-苯氧基苯基)-7-(1-丙醯基哌啶-4-基)-4,5-二氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 81 : 5-oxo-2-(4-phenoxyphenyl)-7-(1-propionylpiperidin-4-yl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide
目標產品由化合物80和丙烯醯氯，參照化合物8的類似方法合成。1H NMR(400MHz,DMSO-d6)δ 11.74(s,1H),7.75(d,J=8.4Hz,2H),7.47-7.41(m,2H),7.20(t,J=7.2Hz,1H),7.12-7.09(m,4H),6.85(dd,J=10.6,16.6Hz,1H),6.12(dd,J=2.4,16.6Hz,1H),5.76(s,1H),5.69(dd,J=2.4,10.6Hz,1H),4.63-4.58(m,1H),4.24-4.20(m,1H),3.15-3.05(m,2H),2.69-2.63(m,1H),1.99-1.91(m,2H),1.61-1.58(m,2H).MS(ESI)m/e[M+1]+ 483.9。 The target product was synthesized from Compound 80 and acrylonitrile, in a similar manner to Compound 8 . 1 H NMR (400MHz, DMSO- d 6) δ 11.74 (s, 1H), 7.75 (d, J = 8.4Hz, 2H), 7.47-7.41 (m, 2H), 7.20 (t, J = 7.2Hz, 1H ), 7.12-7.09 (m, 4H), 6.85 (dd, J = 10.6, 16.6 Hz, 1H), 6.12 (dd, J = 2.4, 16.6 Hz, 1H), 5.76 (s, 1H), 5.69 (dd, J = 2.4, 10.6 Hz, 1H), 4.63-4.58 (m, 1H), 4.24-4.20 (m, 1H), 3.15-3.05 (m, 2H), 2.69-2.63 (m, 1H), 1.99-1.91 ( m, 2H), 1.61-1.58 (m, 2H). MS (ESI) m/e [M+1] + 483.9.
實施例29：化合物82-83的合成Example 29: Synthesis of Compound 82-83
化合物82：2-氧代-6-(4-苯氧基苯基)-1,2-二氫螺[咪唑並[1,2-b]吡唑-3,4'-哌啶]-7-甲醯胺 Compound 82 : 2-oxo-6-(4-phenoxyphenyl)-1,2-dihydrospiro[imidazo[1,2-b]pyrazole-3,4'-piperidine]-7 -Procarbamide
步驟1：1'-苄基-2-氧代-6-(4-苯氧基苯基)-1,2-二氫螺[咪唑並[1,2-b]吡唑-3,4'-哌啶]-7-甲腈 Step 1:1 '-Benzyl-2-oxo-6-(4-phenoxyphenyl)-1,2-dihydrospiro[imidazo[1,2-b]pyrazole-3,4' - piperidine]-7-carbonitrile
1-苄基-4-肼基-4'-甲酸乙酯基哌啶鹽酸鹽(350mg,1.0mmol)，2-(甲氧基(4-苯氧基苯基)亞甲基)丙二腈(276mg，1.0mmol)和K2CO3(414mg,3.0mmol)在MeOH(20mL)中的混合物，加熱回流16小時。混合物過濾，濾液濃縮，得到黃色固體粗品(280mg,58.9%)。MS(ESI,m/e)[M+1]+ 475.9。 1-benzyl-4-indolyl-4'-ethyl ester piperidine hydrochloride (350 mg, 1.0 mmol), 2-(methoxy(4-phenoxyphenyl)methylene)propane carbonitrile (276mg, 1.0mmol) and K 2 CO 3 (414mg, 3.0mmol ) in the mixture (20mL) MeOH, heated to reflux for 16 hours. The mixture was filtered and the filtrate was evaporated to purified crystals MS (ESI, m/e) [M+1] + 475.9.
步驟2：1'-苄基-2-氧代-6-(4-苯氧基苯基)-1,2-二氫螺[咪唑並[1,2-b]吡唑-3,4'-哌啶]-7-甲醯胺 Step 2: 1'-Benzyl-2-oxo-6-(4-phenoxyphenyl)-1,2-dihydrospiro[imidazo[1,2-b]pyrazole-3,4' - piperidine]-7-formamide
1'-苄基-2-氧代-6-(4-苯氧基苯基)-1,2-二氫螺[咪唑並[1,2-b]吡唑-3,4'-哌啶]-7-甲腈(200mg,0.42mmol)的H3PO4(15mL)溶液，120℃加熱2小時。將溶液倒入水(10mL)中，EA(10mL×3)萃取。合併有機相，Na2SO4乾燥，濃縮，得到類白色固體粗品(120mg,58.0%)。MS(ESI,m/e)[M+1]+ 493.9。 1'-Benzyl-2-oxo-6-(4-phenoxyphenyl)-1,2-dihydrospiro[imidazo[1,2-b]pyrazole-3,4'-piperidine ] 7-carbonitrile (200mg, 0.42mmol) in H 3 PO 4 (15mL) solution, 120 ℃ for 2 hours. The solution was poured into water (10 mL) and extracted with EA (10 mL×3). The organic phases were combined, Na dried 2 SO 4, and concentrated to give a crude white solid (120mg, 58.0%). MS (ESI, m/e) [M+1] + 493.9.
步驟3：2-氧代-6-(4-苯氧基苯基)-1,2-二氫螺[咪唑並[1,2-b]吡唑3,4'-哌啶]-7-甲醯胺 Step 3: 2-oxo-6-(4-phenoxyphenyl)-1,2-dihydrospiro[imidazo[1,2-b]pyrazole 3,4'-piperidine]-7- Formamide
向1'-苄基-2-氧代-6-(4-苯氧基苯基)-1,2-二氫螺[咪唑並[1,2-b]吡唑-3,4'-哌啶]-7-甲醯胺(120mg,0.24mmol)的MeOH(10mL)溶液中，加入10% w/w Pd(OH)2/C(5mg)。混合物在氫氣環境下攪拌16小時，過濾，濾液濃縮，得到黃色固體粗品(280mg,58.9%)。MS(ESI,m/e)[M+1]+ 403.9。 To 1'-benzyl-2-oxo-6-(4-phenoxyphenyl)-1,2-dihydrospiro[imidazo[1,2-b]pyrazole-3,4'-per To a solution of pyridine]-7-carboxamide (120 mg, 0.24 mmol) in MeOH (10 mL), 10% w/w Pd (OH) 2 / C (5 mg). The mixture was stirred with EtOAc (EtOAc)EtOAc. MS (ESI, m/e) [M+1] + 403.9.
化合物83：1'-丙烯醯基-2-氧代-6-(4-苯氧基苯基)-1,2-二氫螺[咪唑並[1,2-b]吡唑-3,4'-哌啶]-7-甲醯胺 Compound 83 : 1'-propenyl-2-oxo-6-(4-phenoxyphenyl)-1,2-dihydrospiro[imidazo[1,2-b]pyrazole-3,4 '-Piperidine]-7-formamide
目標產品由化合物82和丙烯醯氯，參照化合物8的類似方法合成。1H NMR(400MHz,DMSO-d6)δ 11.91(s,1H),7.71(d,J=8.4Hz,2H),7.47-7.37(m,2H),7.25(br s,1H),7.17(t,J=7.6Hz,1H),7.08-7.02(m,4H),6.88(dd,J=16.6,10.4Hz,1H),6.80(br s,1H),6.16(d,J=16.6Hz,1H),5.72(d,J=10.4Hz,1H),4.32-4.14(m,1H),4.12-3.99(m,1H),3.97-3.81(m,1H),3.76-3.60(m,1H),1.86-1.91(m,4H).MS(ESI,m/e)[M+1]+ 457.9。 The target product was synthesized from Compound 82 and propylene sulfonium chloride in a similar manner to Compound 8 . 1 H NMR (400MHz, DMSO- d 6) δ 11.91 (s, 1H), 7.71 (d, J = 8.4Hz, 2H), 7.47-7.37 (m, 2H), 7.25 (br s, 1H), 7.17 ( t, J = 7.6 Hz, 1H), 7.08-7.02 (m, 4H), 6.88 (dd, J = 16.6, 10.4 Hz, 1H), 6.80 (br s, 1H), 6.16 (d, J = 16.6 Hz, 1H), 5.72 (d, J = 10.4 Hz, 1H), 4.32-4.14 (m, 1H), 4.12-3.99 (m, 1H), 3.97-3.81 (m, 1H), 3.76-3.60 (m, 1H) , 1.86-1.91 (m, 4H). MS (ESI, m/e) [M+1] + 457.9.
實施例30：化合物84-85的合成Example 30: Synthesis of Compound 84-85
化合物84：6-氨基-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 84 : 6-Amino-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide
步驟1：6-硝基-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈 Step 1: 6-Nitro-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile
向5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈(83mg,0.3mmol)的HOAc(2mL)溶液中加入硝基丙二醛鈉(47mg,0.3mmol)。室溫攪拌1小時後，加水(2mL)。將混合物在EA (25mL)和飽和食鹽水(25mL)中分配。合併有機相，飽和食鹽水(25mL×2)洗滌，Na2SO4乾燥並濃縮，得到黃色固體6-硝基-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈(90mg,84%)。 MS(ESI)m/e[M+1]+ 358.2。 To a solution of 5-amino-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitrile (83 mg, 0.3 mmol) in HOAc (2 mL), sodium nitromalonaldehyde (47 mg, 0.3) Mm). After stirring at room temperature for 1 hour, water (2 mL) was added. The mixture was partitioned between EA (25 mL) and brine (25 mL). The combined organic phases with saturated brine (25mL × 2), dried Na 2 SO 4 and concentrated to give a yellow solid 6-nitro-2- (4-phenoxyphenyl) pyrazolo [1,5-a Pyrimidine-3-carbonitrile (90 mg, 84%). MS (ESI) m / e [ M + 1] + 358.2.
步驟2：6-硝基-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈 Step 2: 6-Nitro-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carbonitrile
室溫下，向6-硝基-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈(90mg,0.25mmol)的乙醇(2mL)和DCM(2mL)的溶液中，加入NaBH4(19mg,0.5mmol)。室溫攪拌30分鐘後，加入水(5mL)。將該混合物濃縮，殘餘物在DCM(50mL)和飽和食鹽水(50mL)中分配。合併有機相，飽和食鹽水(50mL×2)洗滌，Na2SO4乾燥並濃縮，得到黃色固體6-硝基-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈(50mg,55%)。MS(ESI)m/e[M+1]+ 362.1。 To 6-nitro-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile (90 mg, 0.25 mmol) in ethanol (2 mL) and DCM (2mL) was added NaBH 4 (19mg, 0.5mmol). After stirring at room temperature for 30 minutes, water (5 mL) was added. The mixture was concentrated and the residue was crystalljjjjjjjjj The organic phase was combined, washed with saturated brine (50 mL×2), dried over Na 2 SO 4 and concentrated to give 6-nitro-2-(4-phenoxyphenyl)-4,5,6,7- Tetrahydropyrazolo[1,5-a]pyrimidine-3-carbonitrile (50 mg, 55%). MS (ESI) m / e [M+1] + 3621.
步驟3：6-氨基-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈 Step 3: 6-Amino-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carbonitrile
向6-硝基-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈(600mg,1.67mmol)的甲醇(30mL)和DCM(10mL)溶液中，加入10% w/w Pd/C(100mg)。該混合物在氫氣條件下，室溫攪拌2小時，過濾。將濾液濃縮，矽膠層析柱純化，PE/EA為洗脫劑，得到白色固體6-氨基-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈(200mg,36%)。MS(ESI)m/e[M+1]+ 332.1。 To 6-nitro-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carbonitrile (600 mg, 1.67 mmol) A solution of methanol (30 mL) and DCM (10 mL) was added 10% w/w Pd/C (100 mg). The mixture was stirred under a hydrogen atmosphere at room temperature for 2 hours and filtered. The filtrate was concentrated, purified on a silica gel column, and eluted with PE/EA to give 6-amino-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazole as a white solid. [1,5-a]pyrimidine-3-carbonitrile (200 mg, 36%). MS (ESI) m / e [M+1] + 3321.
步驟4：6-氨基-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 4: 6-Amino-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide
目標產品由6-氨基-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈，參照化合物2的步驟2的類似方法合成。1H NMR(400MHz,DMSO-d6)δ 7.53-7.48(m,2H),7.46-7.40(m,2H),7.22-7.16(m,1H),7.11-7.03(m,4H),6.58(br s,1H),4.15-4.08(m,1H),3.72-3.67(m,1H),3.40-3.30(m,2H),3.06-2.98(m,1H).MS(ESI)m/e[M+1]+ 350.2。 The target product consists of 6-amino-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carbonitrile, with reference to compound 2 A similar method of step 2 is synthesized. 1 H NMR (400MHz, DMSO- d 6) δ 7.53-7.48 (m, 2H), 7.46-7.40 (m, 2H), 7.22-7.16 (m, 1H), 7.11-7.03 (m, 4H), 6.58 ( Br s, 1H), 4.15-4.08 (m, 1H), 3.72-3.67 (m, 1H), 3.40-3.30 (m, 2H), 3.06-2.98 (m, 1H). MS (ESI) m/e [ M+1] + 350.2.
化合物85：6-丙烯醯胺基-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 85 : 6-Acrylamino-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide
目標產品由化合物84和丙烯醯氯，參照化合物8的類似方法合成。1H NMR(400MHz,DMSO-d6)δ 8.47(d,J=7.2Hz,1H),7.52(d,J=8.8Hz,2H),7.46-7.38(m,2H),7.18(dd,J=7.2,7.6Hz,1H),7.09(d,J=8.0Hz,2H),7.05(d,J=8.8Hz,2H),6.67(br s,1H),6.34(dd,J=10.0,17.2Hz,1H),6.15(dd,J=2.0,17.2Hz,1H),5.63(dd,J=2.0,10.0Hz,1H),4.32-4.40(m,1H),4.22(dd,J=4.8,12.4Hz,1H),3.91(dd,J=4.8,12.4Hz,1H),3.40(m,1H),3.26(dd,J=5.2,12.0Hz,1H).MS(ESI)m/e[M+1]+ 404.1。 The target product was synthesized from a compound 84 and propylene chloride in a similar manner to compound 8 . 1 H NMR (400MHz, DMSO- d 6) δ 8.47 (d, J = 7.2Hz, 1H), 7.52 (d, J = 8.8Hz, 2H), 7.46-7.38 (m, 2H), 7.18 (dd, J =7.2, 7.6 Hz, 1H), 7.09 (d, J = 8.0 Hz, 2H), 7.05 (d, J = 8.8 Hz, 2H), 6.67 (br s, 1H), 6.34 (dd, J =10.0, 17.2 Hz, 1H), 6.15 (dd, J = 2.0, 17.2 Hz, 1H), 5.63 (dd, J = 2.0, 10.0 Hz, 1H), 4.32-4.40 (m, 1H), 4.22 (dd, J = 4.8, 12.4 Hz, 1H), 3.91 (dd, J = 4.8, 12.4 Hz, 1H), 3.40 (m, 1H), 3.26 (dd, J = 5.2, 12.0 Hz, 1H). MS (ESI) m/e [M +1] + 404.1.
實施例31：化合物86的合成Example 31: Synthesis of Compound 86
化合物86：6-(丙烯醯胺基甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 86 : 6-(acrylamidomethyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3- Formamide
步驟1：3-氰基-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-6-羧酸乙酯 Step 1: Ethyl 3-cyano-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-6-carboxylate
向5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈(276mg,1.0mmol)的EtOH(10mL)溶液中，加入2-甲醯基-3-氧代丙酸乙酯(144mg,1.0mmol)和HOAc(5滴)。室溫攪拌16小時後，將混合物過濾。用H2O(10mL×2)洗滌濾餅，乾燥，得到黃色固體3-氰基-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-6-羧酸乙酯(250mg,65%)。MS(ESI)m/e[M+1]+ 384.9。 To a solution of 5-amino-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitrile (276 mg, 1.0 mmol) in EtOH (10 mL) Ethyl propionate (144 mg, 1.0 mmol) and HOAc (5 drops). After stirring at room temperature for 16 hours, the mixture was filtered. The filter cake was washed with H 2 O (10 mL×2) and dried to give a yellow solid, 3-cyano-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-6-carboxylic acid Ethyl ester (250 mg, 65%). MS (ESI) m/e [M+1] + 384.9.
步驟2：6-(羥甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈 Step 2: 6-(Hydroxymethyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carbonitrile
向3-氰基-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-6-羧酸乙酯(250mg,0.65mmol)的DCM(5mL)和CH3OH(5mL)溶液中，加入NaBH4(250mg,6.5mmol)。室溫攪拌16小時後，混合物在DCM/CH3OH(100mL/5mL)和飽和食鹽水(100mL)中分配。將有機層從水層分離，Na2SO4乾燥，並濃縮，得到6-(羥甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈(250mg,100%)。MS(ESI)m/e[M+1]+ 346.9。 3- cyano-2- (4-phenoxyphenyl) pyrazolo [1,5-a] pyrimidine-6-carboxylate (250mg, 0.65mmol) in DCM (5mL) and CH 3 OH (5mL) was added NaBH 4 (250mg, 6.5mmol). After stirring at room temperature for 16 hours, the mixture was partitioned between DCM / CH 3 OH (100 mL / 5 mL) and brine (100 mL). The organic layer was separated from the aqueous layer, dried over Na 2 SO 4 and concentrated to give 6-(hydroxymethyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazole And [1,5-a]pyrimidine-3-carbonitrile (250 mg, 100%). MS (ESI) m/e [M+1] + 346.9.
步驟3：6-(羥甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 3: 6-(Hydroxymethyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide
目標產品由6-(羥甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈，參照化合物2的步驟2的類似方法合成。MS(ESI)m/e[M+1]+ 364.9。 The target product consists of 6-(hydroxymethyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carbonitrile, The synthesis was carried out in a similar manner to that of Step 2 of Compound 2 . MS (ESI) m/e [M+1] + 364.9.
步驟4：6-((1,3-二氧代異二氫吲哚-2-基)甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 4: 6-((1,3-Dioxaisoindoline-2-yl)methyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydro Pyrazolo[1,5-a]pyrimidine-3-carboxamide
向鄰苯二甲醯亞胺(74mg,0.5mmol)的THF(20mL)溶液中，加入PPh3(393mg,1.5mmol)和6-(羥甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺(180mg,0.5mmol)。在0℃，逐滴加入DIAD(253mg,1.25mmol)，攪拌10分鐘。混合物恢復室溫，攪拌16小時。濃縮，矽膠層析柱(5克矽膠，DCM/CH3OH洗脫)純化，得到黃色固體6-((1,3-二氧代異二氫吲哚-2-基)甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺(200mg,62%)。MS(ESI)m/e[M+1]+ 493.9。 The phthaloyl (PEI) (74mg, 0.5mmol) in THF (20mL) was added PPh 3 (393mg, 1.5mmol) and 6- (hydroxymethyl) -2- (4-phenoxyphenyl -4,5,6,7-Tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide (180 mg, 0.5 mmol). DIAD (253 mg, 1.25 mmol) was added dropwise at 0 ° C and stirred for 10 min. The mixture was returned to room temperature and stirred for 16 hours. Concentration, purification by gelatin chromatography column (5 g silica gel, eluting with DCM/CH 3 OH) to give 6-((1,3-dioxoisoindo-2-yl)methyl)-2 as a yellow solid. -(4-Phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide (200 mg, 62%). MS (ESI) m / e [M+1] + 493.9.
步驟5：6-(氨基甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 5: 6-(Aminomethyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide
向6-((1,3-二氧代異二氫吲哚-2-基)甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺(200mg,0.40mmol)的CH3OH(5mL)溶液中，加入水合肼(1mL,80%的水溶液)。混合物在70℃，N2保護下攪拌4小時，濃縮，矽膠層析柱(5克矽膠，DCM/CH3OH洗脫)純化，得到無色油狀物6-(氨基甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺(63mg,43%)。MS(ESI)m/e[M+1]+ 363.9。 To 6-((1,3-dioxoisoindoline-2-yl)methyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazole and [1,5-a] pyrimidine-3-acyl-amine (200mg, 0.40mmol) in CH 3 OH (5mL) was added hydrazine hydrate (1mL, 80% in water). The mixture was stirred at 70 ℃, N 2 protection 4 hours, concentrated, purified by silica gel column chromatography (5 g silica gel, DCM / CH 3 OH elution) to give a colorless oil of 6- (aminomethyl) -2- ( 4-Phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide (63 mg, 43%). MS (ESI) m/e [M+1] + 363.9.
步驟6：6-(丙烯醯胺基甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 6: 6-(Propylguanidinomethyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3- Formamide
目標產品由6-(氨基甲基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺和丙烯醯氯，參照化合物8的類似方法合成。1H NMR(400MHz,DMSO-d6)δ 8.31(t,J=5.6Hz,1H),7.51(d,J=8.8Hz,2H),7.46-7.38(m,2H),7.[7(t,J= 7.6Hz,1H),7.08(d,J=7.6Hz,2H),7.04(d,J=8.8Hz,2H),6.61(s,1H),6.25(dd,J=17.1,10.1Hz,1H),6.11(dd,J=17.1,2.2Hz,1H),5.62(dd,J=10.1,2.2Hz,1H),4.07(dd,J=12.4,6.0Hz,1H),3.73(dd,J=12.4,8.0Hz,1H),3.41-3.34(m,1H),3.27-3.21(m,2H),3.09-2.96(m,1H),2.37-2.24(m,1H).MS(ESI)m/e[M+1]+ 417.9。 The target product consists of 6-(aminomethyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide It was synthesized in the same manner as in Reference Example 8 with propylene hydrazine chloride. 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.31 (t, J = 5.6 Hz, 1H), 7.51 (d, J = 8.8 Hz, 2H), 7.46-7.38 (m, 2H), 7. [7 ( t, J = 7.6 Hz, 1H), 7.08 (d, J = 7.6 Hz, 2H), 7.04 (d, J = 8.8 Hz, 2H), 6.61 (s, 1H), 6.25 (dd, J = 17.1, 10.1 Hz, 1H), 6.11 (dd, J = 17.1, 2.2 Hz, 1H), 5.62 (dd, J = 10.1, 2.2 Hz, 1H), 4.07 (dd, J = 12.4, 6.0 Hz, 1H), 3.73 (dd , J =12.4, 8.0 Hz, 1H), 3.41-3.34 (m, 1H), 3.27-3.21 (m, 2H), 3.09-2.96 (m, 1H), 2.37-2.24 (m, 1H). MS (ESI ) m/e[M+1]+ 417.9.
實施例32：化合物87-88的合成Example 32: Synthesis of Compound 87-88
化合物87：2'-(4-苯氧基苯基)-5',7'-二氫-4'H-螺[氮雜環丁烷-3,6'-吡唑並[1,5-a]嘧啶]-3'-甲醯胺 Compound 87 : 2'-(4-phenoxyphenyl)-5',7'-dihydro-4'H-spiro[azetidin-3,6'-pyrazolo[1,5- a]pyrimidine]-3'-formamide
步驟1：1-苄基氮雜環丁烷-3,3-二羧酸二乙酯 Step 1: 1-Benzylazetidine-3,3-dicarboxylic acid diethyl ester
在-20℃，向雙羥甲基丙二酸二乙酯(4.4g,20mmol)的CH3CN(50mL)溶液中加入Tf2O(7.1mL,11.85g,42mmol)，然後分兩批加入DIEA(6.45g,50mmol)。0.5小時後，在-20℃，加入苄胺(3.21g,35mmol)。混合物70℃攪拌2小時。加入EA(100mL)和飽和食鹽水(100mL)。有機相Na2SO4乾燥。矽膠層析柱純化，PE/EA洗脫，得到黃色油狀物1-苄基氮雜環丁烷-3,3-二羧酸二乙酯(4.8g,82%)。1H NMR(400MHz, DMSO-d6)δ 7.29-7.34(m,2H),7.22-7.27(m,3H),4.17(q,J=7.2Hz,4H),3.56(s,2H),3.51(s,4H),2.39(s,3H),1.17(t,J=7.2Hz,6H)。 At -20 ℃, the bis-hydroxymethyl-malonate (4.4g, 20mmol) in CH 3 CN (50mL) was added Tf 2 O (7.1mL, 11.85g, 42mmol), was then added in two portions DIEA (6.45 g, 50 mmol). After 0.5 hours, benzylamine (3.21 g, 35 mmol) was added at -20 °C. The mixture was stirred at 70 ° C for 2 hours. EA (100 mL) and saturated brine (100 mL) were added. The organic phase was dried over Na 2 SO 4 . Purification on a silica gel column eluting with EtOAc/EtOAc afforded EtOAc (EtOAc:EtOAc: 1 H NMR (400MHz, DMSO- d 6) δ 7.29-7.34 (m, 2H), 7.22-7.27 (m, 3H), 4.17 (q, J = 7.2Hz, 4H), 3.56 (s, 2H), 3.51 (s, 4H), 2.39 (s, 3H), 1.17 (t, J = 7.2 Hz, 6H).
步驟2：(1-苄基氮雜環丁烷-3,3-二基)二甲醇 Step 2: (1-Benzylazetidin-3,3-diyl)dimethanol
向1-苄基氮雜環丁烷-3,3-二羧酸二乙酯(4.8g,16.5mmol)的CH3OH(10mL)溶液中，加入NaBH4(1.25g,33mmol)。混合物室溫攪拌1小時，加入飽和食鹽水(100mL)和DCM(200ml)。將有機層從水層分離，Na2SO4乾燥，濃縮，得到黃色油狀物(1-苄基氮雜環丁烷-3,3-二基)二甲醇(2.328g,68%)。1H NMR(400MHz,DMSO-d6)δ 7.21-7.31(m,5H),4.15-4.05(m,2H),3.48(d,J=4.8Hz,2H),3.17(d,J=4.8Hz,4H),2.89(s,2H)。 1-benzyl-azetidine-3,3-dicarboxylic acid diethyl ester (4.8g, 16.5mmol) in CH 3 OH (10mL) was added NaBH 4 (1.25g, 33mmol). The mixture was stirred at room temperature for 1 hour, and brine (100 mL) and DCM (EtOAc) The organic layer was separated from the aqueous layer, dried Na 2 SO 4, and concentrated to give a yellow oil (1-benzyl-3,3-diyl) dimethanol (2.328g, 68%). 1 H NMR (400 MHz, DMSO-d 6 ) δ 7.21-7.31 (m, 5H), 4.15-4.05 (m, 2H), 3.48 (d, J = 4.8 Hz, 2H), 3.17 (d, J = 4.8 Hz , 4H), 2.89 (s, 2H).
步驟3：(1-苄基氮雜環丁烷-3,3-二基)雙(亞甲基)二甲基磺酸酯 Step 3: (1-Benzylazetidin-3,3-diyl)bis(methylene)dimethylsulfonate
向(1-苄基氮雜環丁烷-3,3-二基)二甲醇(50mg,0.24mmol)的DCM(10mL)溶液中，加入TEA(222mg,2.2mmoL)和MsCl(249mg,2.2mmol)。室溫攪拌4小時後，將混合物濃縮。殘餘物在飽和食鹽水(100mL)和EA(100mL)中分配。有機相飽和食鹽水(100mL×2)洗滌，Na2SO4乾燥並濃縮，得到 黃色油狀物粗品(1-苄基氮雜環丁烷-3,3-二基)雙(亞甲基)二甲基磺酸酯(300mg,83%)。MS(ESI)m/e[M+1]+ 363.9。 To a solution of (1-benzylazetidin-3,3-diyl)dimethanol (50 mg, 0.24 mmol) in EtOAc (EtOAc) ). After stirring at room temperature for 4 hours, the mixture was concentrated. The residue was partitioned between saturated brine (100 mL) and EtOAc. Washing, drying the organic phase with saturated brine (100mL × 2) Na 2 SO 4 and concentrated to give a yellow oil crude (1-benzyl-3,3-diyl) bis (methylene) Dimethyl sulfonate (300 mg, 83%). MS (ESI) m/e [M+1] + 363.9.
步驟4：1-苄基-2'-(4-苯氧基苯基)-5',7'-二氫-4'H-螺[氮雜環丁烷-3,6'-吡唑並[1,5-a]嘧啶]-3'-甲腈 Step 4: 1-Benzyl-2'-(4-phenoxyphenyl)-5',7'-dihydro-4'H-spiro[azetidin-3,6'-pyrazole [1,5-a]pyrimidin]-3'-carbonitrile
向(1-苄基氮雜環丁烷-3,3-二基)雙(亞甲基)二甲基磺酸酯(300mg,0.83mmol)的DMF(10mL)溶液中，加入5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈(230mg,0.83mmol)和K2CO3(230mg,1.66mmol)。在N2保護下，混合物80℃攪拌16小時。將混合物濃縮，殘餘物用H2O(100mL×2)洗滌，乾燥，並通過製備矽膠板(DCM/CH3OH=10/1)純化，得到黃色液體產品(30mg,10%)。MS(ESI)m/e[M+1]+ 447.9。 To a solution of (1-benzylazetidin-3,3-diyl)bis(methylene)dimethylsulfonate (300 mg, 0.83 mmol) in DMF (10 mL) 3- (4-phenoxyphenyl) lH-pyrazole-4-carbonitrile (230mg, 0.83mmol) and K 2 CO 3 (230mg, 1.66mmol ). The mixture was stirred at 80 ° C for 16 hours under N 2 protection. The mixture was concentrated, the residue was purified by preparative plate silica gel (DCM / CH 3 OH = 10 /1) to give a yellow liquid product (30mg, 10%) with H 2 O (100mL × 2), dried, and. MS (ESI) m / e [M+1] + 447.9.
步驟5：1-苄基-2'-(4-苯氧基苯基)-5',7'-二氫-4'H-螺[氮雜環丁烷-3,6'-吡唑並[1,5-a]嘧啶]-3'-甲醯胺 Step 5: 1-Benzyl-2'-(4-phenoxyphenyl)-5',7'-dihydro-4'H-spiro[azetidin-3,6'-pyrazole [1,5-a]pyrimidine]-3'-formamide
目標產品由1-苄基-2'-(4-苯氧基苯基)-5',7'-二氫-4'H-螺[氮雜環丁烷-3,6'-吡唑並[1,5-a]嘧啶]-3'-甲腈，參照化合物2的步驟2的類似方法合成。MS(ESI)m/e[M+1]+ 465.9。 The target product consists of 1-benzyl-2'-(4-phenoxyphenyl)-5',7'-dihydro-4'H-spiro[azetidin-3,6'-pyrazole [1,5-a]pyrimidin]-3'-carbonitrile was synthesized in a similar manner to that of Step 2 of Compound 2 . MS (ESI) m/e [M+1] + 465.9.
步驟6：2'-(4-苯氧基苯基)-5',7'-二氫-4'H-螺[氮雜環丁烷-3,6'-吡唑並[1,5-a]嘧啶]-3'-甲醯胺 Step 6: 2'-(4-Phenoxyphenyl)-5',7'-dihydro-4'H-spiro[azetidin-3,6'-pyrazolo[1,5- a]pyrimidine]-3'-formamide
向1-苄基-2'-(4-苯氧基苯基)-5',7'-二氫-4'H-螺[氮雜環丁烷-3,6'-吡唑並[1,5-a]嘧啶]-3'-甲醯胺(200mg,0.43mmol)的DCM(10mL)和CH3OH(10mL)溶液中，加入10% w/w Pd/C(100mg)。在H2條件下，室溫攪拌16小時後，將混合物過濾，濃縮。將殘餘物通過製備HPLC純化，水相含0.1% TFA，25%到90% CH3CN梯度洗脫，收集合併目標產品，過夜凍幹，得到白色固體產品(30mg,19%)。1H NMR(400MHz,DMSO-d6)δ 8.94(br s,1H),8.84(br s,1H),7.50(d,J=8.4Hz,2H),7.46-7.38(m,2H),7.18(t,J=8.0Hz,1H),7.08(d,J=8.0Hz,2H),7.05(d,J=8.4Hz,2H),6.81(br s,1 H),4.23(s,2 H),3.90-4.00(m,2 H),3.78-3.87(m,2 H),3.47(s,2H).MS(ESI)m/e[M+1]+ 375.9。 To 1-benzyl-2'-(4-phenoxyphenyl)-5',7'-dihydro-4'H-spiro[azetidin-3,6'-pyrazolo[1] , 5-a] pyrimidine] -3'-acyl amine (200mg, 0.43mmol) in DCM (10mL) and CH 3 OH (10mL) was added 10% w / w Pd / C (100mg). After stirring at room temperature for 16 hours under H 2 , the mixture was filtered and concentrated. The residue was purified by preparative HPLC, an aqueous phase containing 0.1% TFA, 25% to 90% CH 3 CN gradient, to collect the target product were combined, lyophilized overnight, to give the product as a white solid (30mg, 19%). 1 H NMR (400MHz, DMSO- d 6) δ 8.94 (br s, 1H), 8.84 (br s, 1H), 7.50 (d, J = 8.4Hz, 2H), 7.46-7.38 (m, 2H), 7.18 (t, J = 8.0 Hz, 1H), 7.08 (d, J = 8.0 Hz, 2H), 7.05 (d, J = 8.4 Hz, 2H), 6.81 (br s, 1 H), 4.23 (s, 2 H) ), 3.90-4.00 (m, 2 H), 3.78-3.87 (m, 2 H), 3.47 (s, 2H). MS (ESI) m/e [M+1] + 375.9.
化合物88：1-丙烯醯基-2'-(4-苯氧基苯基)-5',7'-二氫-4'H-螺[氮雜環丁烷-3,6'-吡唑並[1,5-a]嘧啶]-3'-甲醯胺 Compound 88 : 1-propenylfluorenyl-2'-(4-phenoxyphenyl)-5',7'-dihydro-4'H-spiro[azetidin-3,6'-pyrazole And [1,5-a]pyrimidin]-3'-formamide
目標產品由化合物87和丙烯醯氯，參照化合物8的類似方法合成。1H NMR(400MHz,CD3OD-d4)δ 7.49(d,J=7.6Hz,2H),7.41-7.32(m,2H),7.14(t,J=8.0Hz,1H),7.05(d,J=8.0Hz,2H),7.04(d,J=7.6Hz,2H),6.35(dd,J=16.8,10.1Hz,1H),6.25(dd,J=16.8,1.6Hz,1H),5.74(dd,J=10.1,1.6Hz,1H),4.20-4.27(m,4 H),3.92-3.98(m,2 H),3.54(s,2 H).MS(ESI)m/e[M+1]+ 429.9。 The target product was synthesized by a similar method of Compound 87 and propylene sulfonium chloride, with reference to Compound 8 . 1 H NMR (400 MHz, CD 3 OD-d 4 ) δ 7.49 (d, J = 7.6 Hz, 2H), 7.41 - 7.32 (m, 2H), 7.14 (t, J = 8.0 Hz, 1H), 7.05 (d) , J = 8.0 Hz, 2H), 7.04 (d, J = 7.6 Hz, 2H), 6.35 (dd, J = 16.8, 10.1 Hz, 1H), 6.25 (dd, J = 16.8, 1.6 Hz, 1H), 5.74 (dd, J =10.1, 1.6 Hz, 1H), 4.20-4.27 (m, 4 H), 3.92-3.98 (m, 2 H), 3.54 (s, 2 H). MS (ESI) m/e [M +1] + 429.9.
實施例33：化合物89-90的合成Example 33: Synthesis of Compound 89-90
化合物89：6-(2-氨基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 89: 6-(2-Aminophenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-A Guanamine
步驟1：6-溴-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈 Step 1: 6-Bromo-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile
5-氨基-3-(4-苯氧基苯基)-1H-吡唑-4-甲腈(28mg,0.1mmol)和2-溴丙二醛(15mg,0.1mmol)在EtOH(5mL)中的混合物，室溫攪拌2小時。然後過濾混合物，得到黃色固體粗品(20mg,62.9%)。1H NMR(400MHz,DMSO-d6)δ 9.89(d,J=2.0Hz,1H),8.95(d,J=2.0Hz,1H),8.08(d,J=8.4Hz,2H),7.52-7.43(m,2H),7.27-7.19(m,3H),7.15(d,J=7.6Hz,2H).MS(ESI,m/e)[M+1]+ 391.9。 5-amino-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitrile (28 mg, 0.1 mmol) and 2-bromomalonaldehyde (15 mg, 0.1 mmol) in EtOH (5 mL) The mixture was stirred at room temperature for 2 hours. The mixture was then filtered to give a crude yellow solid (20mg, 62.9%). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.89 (d, J = 2.0 Hz, 1H), 8.95 (d, J = 2.0 Hz, 1H), 8.08 (d, J = 8.4 Hz, 2H), 7.52 7.43 (m, 2H), 7.27-7.19 (m, 3H), 7.15 (d, J = 7.6 Hz, 2H). MS (ESI, m/e) [M+1] + 391.9.
步驟2：6-(2-氨基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈 Step 2: 6-(2-Aminophenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile
氮氣保護下，6-溴-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈(500mg,1.28mmol)，2-氨基苯硼酸(175mg,1.28mmol)，碳酸銫(623mg,1.92mmol)和Pd(PPh3)4(74mg,0.06mmol)在1,4-二氧六環(30mL)和水(1.0mL)中的混合物，加熱到80℃，攪拌16小時。將混合物過濾，濃縮濾液，矽膠層析柱純化，含50% EA的PE洗脫，得到黃色固體粗品(320mg,59.1%)。MS(ESI,m/e)[M+1]+ 403.9。 6-bromo-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile (500 mg, 1.28 mmol), 2-aminophenylboronic acid (175 mg, under N2) 1.28 mmol), a mixture of cesium carbonate (623 mg, 1.92 mmol) and Pd(PPh 3 ) 4 (74 mg, 0.06 mmol) in 1,4-dioxane (30 mL) and water (1.0 mL). Stir for 16 hours at °C. The mixture was filtered, and the filtrate was evaporated. mjjjjjjjj MS (ESI, m/e) [M+1] + 403.9.
步驟3：6-(2-氨基苯基)-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]嘧啶-3-甲腈 Step 3: 6-(2-Aminophenyl)-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]pyrimidine-3-carbonitrile
向6-(2-氨基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈(320mg,0.79mmol)的MeOH(20mL)溶液中，加入NaBH4(86mg,2.28mmol)。混合物室溫攪拌30分鐘，然後倒入水(50mL)中，並用EA(50mL×3)萃取。合併有機相，Na2SO4乾燥，濃縮得到黃色固體粗品(240mg,75%)。MS(ESI,m/e)[M+1]+ 406.0。 To a solution of 6-(2-aminophenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile (320 mg, 0.79 mmol) in MeOH (20 mL) NaBH 4 (86 mg, 2.28 mmol) was added. The mixture was stirred at room temperature for 30 min then poured into water (50 mL) and EtOAc (EtOAc) The organic phases were combined, Na dried 2 SO 4, and concentrated to give the crude product as a yellow solid (240mg, 75%). MS (ESI, m/e) [M+1] + 406.0.
步驟4：6-(2-氨基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈 Step 4: 6-(2-Aminophenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-A Nitrile
目標產物由6-(2-氨基苯基)-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]嘧啶-3-甲腈，根據化合物68的步驟8的類似方法合成。MS(ESI,m/e)[M+1]+ 407.9。 The target product consists of 6-(2-aminophenyl)-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]pyrimidine-3-carbonitrile, according to the compound A similar method of step 8 of 68 is synthesized. MS (ESI, m/e) [M+1] + 407.9.
步驟5：6-(2-氨基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 5: 6-(2-Aminophenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-A Guanamine
目標產品由6-(2-氨基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈，參照化合物2的步驟2的類似方法合成。1H NMR(400MHz,DMSO-d6)δ 7.58(d,J=8.4Hz,2H),7.53-7.44(m,2H),7.24(t,J=7.6Hz,1H),7.19-7.09(m,4H),7.06-7.99(m,2H),6.84(d,J=1.6Hz,1H),6.74(d,J=7.2Hz,1H),6.61(t,J=7.6Hz,1H),5.20(s,2H),4.24(t,J=12.0,Hz,1H),4.07(t,J=12.0Hz,1H),3.56-3.41(m,3H).MS(ESI,m/e)[M+1]+ 425.9。 The target product consists of 6-(2-aminophenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-methyl The nitrile was synthesized in a similar manner to that of Step 2 of Compound 2 . 1 H NMR (400 MHz, DMSO-d 6 ) δ 7.58 (d, J = 8.4 Hz, 2H), 7.53-7.44 (m, 2H), 7.24 (t, J = 7.6 Hz, 1H), 7.19-7.09 (m) , 4H), 7.06-7.99 (m, 2H), 6.84 (d, J = 1.6 Hz, 1H), 6.74 (d, J = 7.2 Hz, 1H), 6.61 (t, J = 7.6 Hz, 1H), 5.20 (s, 2H), 4.24 (t, J = 12.0, Hz, 1H), 4.07 (t, J = 12.0 Hz, 1H), 3.56-3.41 (m, 3H). MS (ESI, m/e) [M +1] + 425.9.
化合物90：6-(3-氨基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 90: 6-(3-Aminophenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-A Guanamine
根據化合物89的步驟2到5製備方案，在本領域普通技術人員熟知的條件下，以6-溴-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈和3-氨基苯硼酸為起始原料，得到目標產物。1H NMR(400MHz,DMSO-d6)δ 7.59(d,J=8.4Hz,2H),7.52-7.45(m,2H),7.24(t,J=7.6Hz,1H),7.15(d,J=8.4Hz,2H),7.12(d,J=7.6Hz,2H),7.07(t,J=7.6Hz,1H),6.85(br s, 1H),6.62-6.54(m,3H),5.23(br s,2H),4.24(dd,J=12.0,4.8Hz,1H),4.09(t,J=12.0Hz,1H),3.56-3.50(m,1H),3.40-3.35(m,1H),3.28-3.19(m,1H).MS(ESI,m/e)[M+1]+ 425.9。 According to the preparative scheme of steps 2 to 5 of compound 89 , 6-bromo-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine is used under conditions well known to those skilled in the art. 3-carbonitrile and 3-aminobenzeneboronic acid are used as starting materials to obtain the desired product. 1 H NMR (400MHz, DMSO- d 6) δ 7.59 (d, J = 8.4Hz, 2H), 7.52-7.45 (m, 2H), 7.24 (t, J = 7.6Hz, 1H), 7.15 (d, J = 8.4 Hz, 2H), 7.12 (d, J = 7.6 Hz, 2H), 7.07 (t, J = 7.6 Hz, 1H), 6.85 (br s, 1H), 6.62 - 6.54 (m, 3H), 5.23 ( Br s, 2H), 4.24 (dd, J = 12.0, 4.8 Hz, 1H), 4.09 (t, J = 12.0 Hz, 1H), 3.56-3.50 (m, 1H), 3.40-3.35 (m, 1H), 3.28-3.19 (m, 1H). MS (ESI, m/e) [M+1] + 425.9.
實施例34：化合物91的合成Example 34: Synthesis of Compound 91
化合物91：6-(3-(2-(二甲基氨基)乙氧基)苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 91 : 6-(3-(2-(Dimethylamino)ethoxy)phenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazole [1,5-a]pyrimidine-3-carboxamide
步驟1：6-(3-羥基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈 Step 1: 6-(3-Hydroxyphenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile
向6-溴-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈(782mg,2.0mmol)的二氧六環(10mL)和水(10mL)的溶液中，加入3-羥基苯硼酸(276mg,2.0mmol)，Pd(PPh3)4(240mg,0.2mmol)和Na2CO3(424mg,4.0mmol)。在氮氣保護下，65℃攪拌16小時後，濃縮該混合物，加入DCM(100mL)，CH3OH(10mL)和水(100mL)。將有機層從水層分離， Na2SO4乾燥，矽膠層析柱純化，二氯甲烷/甲醇洗脫，得到黃色固體6-(3-羥基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈(500mg,62%)。MS(ESI)m/e[M+1]+ 404.9。 To 6-bromo-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile (782 mg, 2.0 mmol) in dioxane (10 mL) and water (10 mL) To the solution, 3-hydroxybenzeneboronic acid (276 mg, 2.0 mmol), Pd(PPh 3 ) 4 (240 mg, 0.2 mmol) and Na 2 CO 3 (424 mg, 4.0 mmol) were added. Under nitrogen. After stirring for 65 ℃ 16 h, the mixture was concentrated, DCM (100mL), CH 3 OH (10mL) and water (100mL). The organic layer was separated from the aqueous layer, dried Na 2 SO 4, purified by column chromatography on silica gel, dichloromethane / methanol to give a yellow solid 6- (3-hydroxyphenyl) -2- (4-phenoxy-benzene Pyrazolo[1,5-a]pyrimidine-3-carbonitrile (500 mg, 62%). MS (ESI) m/e [M+1] + 404.9.
步驟2：6-(3-羥基苯基)-2-(4-苯氧基苯基)-6,7-二氫吡唑並[1,5-a]嘧啶-3-甲腈和6-(3-羥基苯基)-2-(4-苯氧基苯基)-4,5-二氫吡唑並[1,5-a]嘧啶-3-甲腈 Step 2: 6-(3-Hydroxyphenyl)-2-(4-phenoxyphenyl)-6,7-dihydropyrazolo[1,5-a]pyrimidine-3-carbonitrile and 6- (3-hydroxyphenyl)-2-(4-phenoxyphenyl)-4,5-dihydropyrazolo[1,5-a]pyrimidine-3-carbonitrile
目標產物由6-(3-羥基苯基)-2-(4-苯氧基苯基)吡唑並[1,5-a]嘧啶-3-甲腈，根據化合物89的步驟3的類似方法合成。MS(ESI)m/e[M+1]+ 406.9。 The desired product consists of 6-(3-hydroxyphenyl)-2-(4-phenoxyphenyl)pyrazolo[1,5-a]pyrimidine-3-carbonitrile, analogously to Step 3 of Compound 89 synthesis. MS (ESI) m/e [M+1] + 406.9.
步驟3：6-(3-羥基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈 Step 3: 6-(3-Hydroxyphenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-A Nitrile
目標產物由上步中間體，根據化合物89的步驟4的類似方法合成。MS(ESI)m/e[M+1]+ 406.9。MS(ESI)m/e[M+1]+ 408.9。 The title product was synthesized from the above intermediate, which was obtained in a similar manner to step 4 of compound 89 . MS (ESI) m/e [M+1] + 406.9. MS (ESI) m/e [M+1] + 408.9.
步驟4：6-(3-羥基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 4: 6-(3-Hydroxyphenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-A Guanamine
目標產品由6-(3-羥基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲腈，參照化合物2的步驟2合成。MS(ESI)m/e[M+1]+ 426.9。 The target product consists of 6-(3-hydroxyphenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-methyl The nitrile was synthesized in the same manner as in the second step of the compound 2 . MS (ESI) m / e [ M + 1] + 426.9.
步驟5：6-(3-(2-(二甲基氨基)乙氧基)苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 5: 6-(3-(2-(Dimethylamino)ethoxy)phenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazole [1,5-a]pyrimidine-3-carboxamide
目標產品以6-(3-羥基苯基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺和二甲氨基氯乙烷鹽酸鹽為起始原料，合成方法與化合物9的步驟7類似。1H NMR(400MHz,DMSO-d6)δ 7.52(d,J=8.4Hz,2H),7.46-7.38(m,2H),7.29(t,J=7.6Hz,1H),7.18(t,J=7.6Hz,1H),7.10-7.05(m,4H),6.98-6.96(m,2H),6.90-6.88(m,1H),6.81(d,J=2.4Hz,1H),4.25-4.20(m,1H),4.16-4.11(m,3H),3.50-3.47(m,1H),3.43-3.38(m,2H),2.96-2.84(m,2H),2.43(s,6H).MS(ESI)m/e[M+1]+ 497.9。 The target product is 6-(3-hydroxyphenyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-methyl The indoleamine and dimethylaminochloroethane hydrochloride were used as starting materials, and the synthesis method was similar to that of Step 7 of Compound 9 . 1 H NMR (400MHz, DMSO- d6) δ 7.52 (d, J = 8.4Hz, 2H), 7.46-7.38 (m, 2H), 7.29 (t, J = 7.6Hz, 1H), 7.18 (t, J = 7.6 Hz, 1H), 7.10-7.05 (m, 4H), 6.98-6.96 (m, 2H), 6.90-6.88 (m, 1H), 6.81 (d, J = 2.4 Hz, 1H), 4.25-4.20 (m) ,1H), 4.16-4.11(m,3H), 3.50-3.47(m,1H),3.43-3.38(m,2H),2.96-2.84(m,2H),2.43(s,6H).MS(ESI )m/e[M+1] + 497.9.
化合物178： N1-(2-(4-((E)-4-(4-((S)-3-氨基甲醯基-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)哌啶-1-基)- 4-氧代丁基-2-烯-1-基)哌嗪-1-基)乙基)-N5-(15-氧代-19-((3aR,4R,6aS)-2-氧代六氫-1H-噻吩並[3,4-d]咪唑-4-基)-4,7,10-三氧雜-14-氮雜十九烷基)戊二醯胺三氟乙酸鹽 Compound 178: N 1-(2-(4-((E)-4-(4-((S)-3-aminocarbamido-2-(4-phenoxyphenyl)-4,5, 6,7-tetrahydropyrazolo[1,5-a]pyrimidin-7-yl)piperidin-1-yl)-4-oxobutyl-2-en-1-ylpiperazin-1- Ethyl) -N 5-(15-oxo-19-((3aR,4R,6aS)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl) -4,7,10-trioxa-14-aza-nonadecyl)pentaneamine trifluoroacetate
目標化合物的製備根據如下所述的圖示，步驟和中間體。 The preparation of the target compound is based on the schemes, procedures and intermediates described below.
步驟1：(E)-4-(4-(2-((叔丁氧基羰基)氨基)乙基)哌嗪-1-基)丁-2-烯酸 Step 1: ( E )-4-(4-(2-((tert-Butoxycarbonyl)amino)ethyl)piperazin-1-yl)but-2-enoic acid
混合物(E)-4-溴丁-2-烯酸(500mg，3.03mmol)，(2-(哌嗪-1-基)乙基)氨基甲酸叔丁酯(694mg，3.03mmol)和Et3N(612mg，6.06mmol)在THF(20mL)中室溫攪拌15h。將混合物濃縮，未經進一步純化直接用於下一步驟。MS(ESI)m/e[M+1]+ 314.0。 Mixture of (E) -4- bromo-2-enoic acid (500mg, 3.03mmol), (2- ( piperazin-1-yl) ethyl) carbamate (694mg, 3.03mmol) and Et 3 N (612 mg, 6.06 mmol) was stirred in THF (20 mL The mixture was concentrated and used in the next step without further purification. MS (ESI) m / e [M+1] + 314.0.
步驟2：(S)-2-(4-苯氧基苯基)-7-(哌啶-4-基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Step 2: (S)-2-(4-Phenoxyphenyl)-7-(piperidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a] Pyrimidine-3-carboxamide
2-(4-苯氧基苯基)-7-(哌啶-4-基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺(2.0g,4.8mmol)在MeOH/H2O=3/1(110mL)的溶液升溫至50℃，攪拌約20分鐘直至溶解所有原料，然後向溶液中加入L-DBTA(600mg,1.6mmol)的MeOH/H2O=3/1(10mL)溶液，將溶液於50℃下攪拌約30分鐘，然後緩慢冷卻至40℃(約2h)。向溶液中加入晶種(10mg)。混合物在40℃下攪拌2h，然後緩慢冷卻至室溫並攪拌約48h。過濾，將固體用MeOH/H2O=3/1(5mL)洗滌，減壓下乾燥得到產物，為白色固體約1.1克(38%的產率，93%的ee值)。該固體(500mg)加入到THF/H2O=1/1(20mL)中，將溶液升溫至70℃攪拌約1小時，直到所有固體溶解。然後緩慢冷卻至40℃(3h)，並加入晶種(10mg)，攪拌約2小時後，將溶液緩慢冷卻至室溫並攪拌約48h。過濾，固體用水(4mL)洗滌，減壓乾燥得到產物的L-DBTA鹽形式，為白色固體330mg(65%的產率，>99.5%的ee值)。通過緩慢冷卻此L-DBTA鹽在MeOH/H2O(1：1,v/v)中的溶液得到合適的單晶。並確定遊離堿 的手性碳的構型為S。該L-DBTA鹽可以通過NaOH水溶液，並用DCM萃取轉化為遊離堿。 2-(4-phenoxyphenyl)-7-(piperidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide (2.0 g, 4.8 mmol) The solution in MeOH / H 2 O = 3 / 1 (110 mL) was warmed to 50 ° C, stirred for about 20 minutes until all the starting materials were dissolved, then L-DBTA (600 mg, 1.6 mmol) was added to the solution. A solution of MeOH/H 2 O = 3/1 (10 mL) was then stirred at 50 ° C for about 30 minutes and then slowly cooled to 40 ° C (about 2 h). Seed crystals (10 mg) were added to the solution. The mixture was stirred at 40 ° C for 2 h, then slowly cooled to room temperature and stirred for about 48 h. Filtered and the solid was washed with MeOH / H 2 O = 3/ 1 (5mL) with, and dried under reduced pressure to give the product as a white solid of about 1.1 grams (38% yield, 93% ee value). The solid (500 mg) was added to THF / H 2 O = 1 / 1 (20 mL), and the solution was warmed to 70 ° C and stirred for about 1 hour until all solids dissolved. It was then slowly cooled to 40 ° C (3 h) and seeded (10 mg) was added. After stirring for about 2 hours, the solution was slowly cooled to room temperature and stirred for about 48 h. Filtration, the solid was washed with water (4 mL) and dried <RTI ID=0.0></RTI> to </RTI><RTIID=0.0></RTI> A suitable single crystal is obtained by slowly cooling a solution of this L-DBTA salt in MeOH/H 2 O (1:1, v/v). It is also determined that the configuration of the chiral carbon of the free ruthenium is S. The L-DBTA salt can be converted to free hydrazine by aqueous NaOH and extraction with DCM.
手性分析條件如下所示：
步驟3：(S,E)-叔丁基(2-(4-(4-(4-(3-氨基甲醯基-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)哌啶-1-基)-4-氧代丁-2-烯-1-基)哌嗪-1-基)乙基)氨基甲酸酯 Step 3: ( S , E )-tert-Butyl (2-(4-(4-(4-(3-aminomethylmethyl)-2-(4-phenoxyphenyl)-4,5,6, 7-Tetrahydropyrazolo[1,5-a]pyrimidin-7-yl)piperidin-1-yl)-4-oxobut-2-en-1-ylpiperazin-1-yl)B Carbamate
混合物(S)-2-(4-苯氧基苯基)-7-(哌啶-4-基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺(1.26g,3.03mmol),(E)-4-(4-(2-((叔丁氧基羰基)氨基)乙基)哌嗪-1-基)丁-2-烯酸(948.4mg,3.03mmol),HATU(1.21g,3.18mmol)和DIEA(782mg,6.06mmol)在DMF(30mL)中室溫攪拌15h。混合物倒入到300mL水中，EA(100mL)萃取。有機相水洗(100mL×3)，濃縮得到1.25g(58%)黃色固體產品。1H NMR(400MHz,DMSO-d 6)δ 7.50(d,J=8.4Hz,2H),7.42(t,J=8.4Hz,2H),7.17(t,J=7.4Hz,1H),7.12-7.02(m,4H),6.67(br s,1H),6.64-6.53(m,3H),4.53-4.40(m,1H),4.14-4.05(m,1H),3.32-3.25(m,2H),3.10-2.91(m,5H),2.45-2.15(m,11H),2.10-2.00(m,1H),1.96-1.84(m,1H),1.78-1.65(m,1H),1.62-1.50(m,1H),1.37(s,9H),1.31-1.10(m,3H).MS(ESI)m/e[M+1]+ 713.0. Mixture (S)-2-(4-phenoxyphenyl)-7-(piperidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine- 3-carbamide (1.26 g, 3.03 mmol), ( E )-4-(4-(2-((tert-butoxycarbonyl)amino)ethyl)piperazin-1-yl)but-2-ene The acid (948.4 mg, 3.03 mmol), EtOAc (EtOAc, m. The mixture was poured into 300 mL of water and extracted with EA (100 mL). The organic phase was washed with water (100 mL x 3) and concentrated to yield 1.25 g ( 58% 1 H NMR (400 MHz, DMSO- d 6 ) δ 7.50 (d, J = 8.4 Hz, 2H), 7.42 (t, J = 8.4 Hz, 2H), 7.17 (t, J = 7.4 Hz, 1H), 7.12 7.02(m,4H),6.67(br s,1H),6.64-6.53(m,3H),4.53-4.40(m,1H),4.14-4.05(m,1H),3.32-3.25(m,2H) , 3.10-2.91 (m, 5H), 2.45-2.15 (m, 11H), 2.10-2.00 (m, 1H), 1.96-1.84 (m, 1H), 1.78-1.65 (m, 1H), 1.62-1.50 ( m, 1H), 1.37 (s, 9H), 1.31-1.10 (m, 3H). MS (ESI) m/e [M+1] + 713.0.
步驟4：(S,E)-7-(1-(4-(4-(2-氨基乙基)哌嗪-1-基)丁-2-烯醯基)哌啶-4-基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺三氟乙酸鹽 Step 4: ( S , E )-7-(1-(4-(4-(2-Aminoethyl)piperazin-1-yl)but-2-enyl)piperidin-4-yl)- 2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide trifluoroacetate
(S,E)-叔丁基(2-(4-(4-(4-(3-氨基甲醯基-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)哌啶-1-基)-4-氧代丁-2-烯-1-基)哌嗪-1-基)乙基)氨基甲酸酯(150mg,0.21mmol)溶於DCM(10mL)，加入2mL的TFA。混合物室溫下攪拌15h，濃縮除去溶劑。殘餘物未經進一步純化直接用於下一步驟。MS(ESI)m/e[M+1]+ 613.0。 ( S , E )-tert-butyl(2-(4-(4-(4-(3-aminomethylmethyl)-2-(4-phenoxyphenyl)-4,5,6,7-tetra Hydropyrazolo[1,5-a]pyrimidin-7-yl)piperidin-1-yl)-4-oxobut-2-en-1-yl)piperazin-1-yl)ethyl)amino Formic acid ester (150 mg, 0.21 mmol) was dissolved in DCM (10 mL). The mixture was stirred at room temperature for 15 h and concentrated and evaporated. The residue was used in the next step without further purification. MS (ESI) m / e [M+1] + 613.0.
化合物178： N1-(2-(4-((E)-4-(4-((S)-3-氨基甲醯基-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-7-基)哌啶-1-基)-4-氧代丁基-2-烯-1-基)哌嗪-1-基)乙基)-N5-(15-氧代-19- ((3aR,4R,6aS)-2-氧代六氫-1H-噻吩並[3,4-d]咪唑-4-基)-4,7,10-三氧雜-14-氮雜十九烷基)戊二醯胺三氟乙酸鹽 Compound 178: N 1-(2-(4-((E)-4-(4-((S)-3-aminocarbamido-2-(4-phenoxyphenyl)-4,5, 6,7-tetrahydropyrazolo[1,5-a]pyrimidin-7-yl)piperidin-1-yl)-4-oxobutyl-2-en-1-ylpiperazin-1- Ethyl) -N 5-(15-oxo-19-((3aR,4R,6aS)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl) -4,7,10-trioxa-14-aza-nonadecyl)pentaneamine trifluoroacetate
混合物(S,E)-7-(1-(4-(4-(2-氨基乙基)哌嗪-1-基)丁-2-烯醯基)哌啶-4-基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺三氟乙酸鹽(129mg,0.21mmol,crude),N-BIOTINYL-NH-(PEG)2-COOH-DIEA(118mg,0.21mmol),HATU(80mg,0.21mmol)和TEA(63.6mg,0.63mmol)在DMF(5mL)中40℃攪拌15h.混合物濃縮，Pre-HPLC純化得到160mg(60%)白色固體產品。1H NMR(400MHz,DMSO-d 6)δ 8.04(t,J=5.2Hz,1H),7.80(t,J=5.6Hz,1H),7.76(t,J=5.6Hz,1H),7.50(d,J=8.5Hz,2H),7.42(t,J=8.5Hz,2H),7.18(t,J=7.6Hz,1H),7.12-7.03(m,4H),6.89-6.77(m,1H),6.62-6.51(m,1H),6.44(s,1H),4.55-4.40(m,1H),4.35-4.27(m,1H),4.17-3.95(m,3H),3.74-3.60(m,2H),3.55-3.44(m,9H),3.41-3.27(m,10H),3.14-3.02(m,9H),2.82(dd,J=12.4,5.0Hz,1H),2.65-2.53(m,3H),2.36-2.18(m,1H),2.13-2.00(m,7H),1.96-1.85(m,1H),1.80-1.67(m,3H),1.66-1.55(m,6H),1.55-1.40(m,3H),1.38-1.21(m,4H).MS(ESI)m/e[M+1]+ 1155.0,[M+23]+ 1176.9. Mixture ( S , E )-7-(1-(4-(4-(2-Aminoethyl)piperazin-1-yl)but-2-enyl)piperidin-4-yl)-2- (4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide trifluoroacetate (129 mg, 0.21 mmol, crude), N-BIOTINYL-NH-(PEG)2-COOH-DIEA (118 mg, 0.21 mmol), EtOAc (EtOAc, EtOAc, EtOAc (EtOAc) Purification by Pre-HPLC gave 160 mg (60%) of a white solid product. 1 H NMR (400MHz, DMSO- d 6) δ 8.04 (t, J = 5.2Hz, 1H), 7.80 (t, J = 5.6Hz, 1H), 7.76 (t, J = 5.6Hz, 1H), 7.50 ( d, J = 8.5 Hz, 2H), 7.42 (t, J = 8.5 Hz, 2H), 7.18 (t, J = 7.6 Hz, 1H), 7.12-7.03 (m, 4H), 6.89-6.77 (m, 1H) ), 6.62-6.51 (m, 1H), 6.44 (s, 1H), 4.55-4.40 (m, 1H), 4.35-4.27 (m, 1H), 4.17-3.95 (m, 3H), 3.74-3.60 (m) , 2H), 3.55-3.44 (m, 9H), 3.41-3.27 (m, 10H), 3.14-3.02 (m, 9H), 2.82 (dd, J = 12.4, 5.0 Hz, 1H), 2.65-2.53 (m , 3H), 2.36-2.18 (m, 1H), 2.13-2.00 (m, 7H), 1.96-1.85 (m, 1H), 1.80-1.67 (m, 3H), 1.66-1.55 (m, 6H), 1.55 - 1.40 (m, 3H), 1.38-1.21 (m, 4H). MS (ESI) m/e [M+1] + 1155.0, [M+23] + 1176.9.
化合物181：(S)-7-(1-(2-氰基-3-環丙基丙烯醯基)哌啶-4-基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 181: (S)-7-(1-(2-Cyano-3-cyclopropylpropenyl)piperidin-4-yl)-2-(4-phenoxyphenyl)-4,5 ,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide
化合物180(60mg,0.124mmol)，環丙基甲醛(43.4mg,0.62mmol)和哌啶(52.7mg,0.62mmol)在MeOH(10mL)中的混合物在室溫下攪拌15h。濃縮後，向殘餘物中加入EA(50mL)和水(50mL)。將有機相濃縮，通過色譜矽膠柱純化，用DCM/MeOH(50/1)洗脫，得到白色固體化合物30mg(45%)。 MS(ESI)m/e[M+1]+ 537.0. A mixture of compound 180 (60 mg, 0.124 mmol), EtOAc (EtOAc) After concentration, EA (50 mL) and water (50 mL). The organic phase was concentrated, purified by EtOAc EtOAc EtOAc EtOAc MS (ESI) m / e [M + 1] + 537.0.
化合物182：(S)-7-(1-氰基哌啶-4-基)-2-(4-苯氧基苯基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺 Compound 182: (S)-7-(1-Cyanopiperidin-4-yl)-2-(4-phenoxyphenyl)-4,5,6,7-tetrahydropyrazolo[1, 5-a]pyrimidine-3-carboxamide
向(S)-2-(4-苯氧基苯基)-7-(哌啶-4-基)-4,5,6,7-四氫吡唑並[1,5-a]嘧啶-3-甲醯胺(417mg,1mmol)的DCM(20mL)溶液中加入NaHCO3(168mg,2mmol)和水(5mL)，BrCN(127mg,1.2mmol)。混合物室溫下攪拌16h。向混合物中加入DCM(50mL)和食鹽水(20mL)。將有機相進一步用食鹽水(100mL)洗滌，Na2SO4乾燥。濃縮，通過Pre-TLC (DCM/MeOH,50/1)純化，得到330mg(75%)白色固體。MS(ESI)m/e[M+1]+ 443.0. To (S)-2-(4-phenoxyphenyl)-7-(piperidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine- 3-acyl amine (417mg, 1mmol) in DCM (20mL) was added NaHCO 3 (168mg, 2mmol) and water (5mL), BrCN (127mg, 1.2mmol). The mixture was stirred at room temperature for 16 h. DCM (50 mL) and brine (20 mL) were added to the mixture. The organic phase further washed with brine (100 mL), dried Na 2 SO 4. Concentration, purification by Pre-TLC (DCM /MeOH, 50/1) MS (ESI) m / e [M + 1] + 443.0.
一些其他化合物的合成大體上與上述實施例的方法類似。表1中給出了這些化合物的表徵資料，其中包括LC/MS(實測的)，手性HPLC和1H NMR資料。 The synthesis of some other compounds is generally similar to the method of the above examples. Characterization data for these compounds are given in Table 1, including LC/MS (measured), chiral HPLC and 1 H NMR data.
BTK激酶檢測 BTK kinase assay
本發明公開的化合物對BTK激酶活性的抑制作用是在時間分辨螢光共振能量轉移的方法裡檢測的。重組的BTK與本發明公開的化合物在含有50mM Tris pH7.4,10mM MgCl2,2mM MnCl2,0.1mM EDTA,1mM DTT,20nM SEB,0.1%BSA,0.005% tween-20的緩衝液中在室溫下預先孵育1個小時。加入ATP(在ATP Km濃度)和多肽底物(Biotin-AVLESEEELYSSARQ-NH2)來啟動反應。室溫孵育一個小時後，等體積的終止液含有50mM HEPES pH7.0,800mM KF,20mM EDTA,0.1% BSA,Eu cryptate-conjugated p-Tyr66 antibody和streptavidin-labeled XL665被加入到反應中去終止反應。板子在室溫下再孵育一個小時，然後在BMG PHERAstar FS儀器上讀取TR-FRET信號(ex337nm,em 620nm/665nm)。在化合物濃度遞增下，剩餘酶的活性通過在615nm與665nm的螢光信號的比值被計算出來。每個化合物的IC50使用Graphpad Prism軟體的四參數方程來計算出來。 The inhibition of BTK kinase activity by the compounds disclosed herein is detected in a time resolved fluorescence resonance energy transfer method. Recombinant BTK and the compound disclosed in the present invention are in a buffer containing 50 mM Tris pH 7.4, 10 mM MgCl 2 , 2 mM MnCl 2 , 0.1 mM EDTA, 1 mM DTT, 20 nM SEB, 0.1% BSA, 0.005% tween-20. Incubate for 1 hour before warming. The reaction was initiated by the addition of ATP (at ATP Km concentration) and the polypeptide substrate (Biotin-AVLESEEELYSSARQ-NH 2 ). After incubation for one hour at room temperature, an equal volume of stop solution containing 50 mM HEPES pH 7.0, 800 mM KF, 20 mM EDTA, 0.1% BSA, Eu cryptate-conjugated p-Tyr66 antibody and streptavidin-labeled XL665 were added to the reaction to stop the reaction. The plates were incubated for an additional hour at room temperature and then the TR-FRET signal (ex337 nm, em 620 nm / 665 nm) was read on a BMG PHERAstar FS instrument. At the increasing concentration of the compound, the activity of the remaining enzyme was calculated by the ratio of the fluorescence signals at 615 nm to 665 nm. The IC50 of each compound was calculated using the four parameter equation of the Graphpad Prism software.
細胞水準BTKpY223檢測技術：細胞水準BTKpY223檢測技術是基於均相時間分辨螢光技術(HTRF)開發的檢測技術，用於檢測內源BTK在223位酪氨酸磷酸化的水準。檢驗所用細胞系為Ramos細胞系(CRL-1596，ATCC)，試劑為BTKpY223檢測試劑盒(63IDC000，Cisbio公司)。 Cell Level BTKpY223 Detection Technology: Cellular Level BTKpY223 detection technology is a detection technique developed based on homogeneous time-resolved fluorescence (HTRF) to detect the level of tyrosine phosphorylation of endogenous BTK at position 223. The cell line used for the assay was the Ramos cell line (CRL-1596, ATCC) and the reagent was the BTKpY223 assay kit (63 IDC000, Cisbio).
簡單地描述過程如下，Ramos細胞先在含有0.5%胎牛血 清的RPMI1640培養基中血清饑餓培養2小時。之後，細胞會與不同濃度的化合物混合，並在CO2培養箱中孵育1小時。孵育結束後，細胞用1毫摩釩酸鈉或者過釩酸鈉刺激20分鐘。然後，離心收集細胞，細胞與1倍濃度的細胞裂解液(試劑盒中提供了4倍濃度的儲液)在室溫(25攝氏度)孵育10分鐘以裂解細胞。孵育期間，配製抗體混合液，把BTK-d2抗體和pBTK-K抗體稀釋於檢測緩衝液中(試劑盒中提供)。在OptiPlate-384檢測板(6005620，PerkinElmer公司)中，每個微孔中加2微升抗體混合液。然後，把18微升裂解細胞後的細胞裂解液轉移到之前加入抗體混合液的微孔中。輕柔混勻後短暫離心，用封板膜封閉後置於避光處室溫孵育18小時。孵育結束後，在讀板機(PHERAstar FS,BMG公司)中測量665nm和620nm兩個波長的發射螢光。化合物的活性計算是基於對665nm和620nm處信號強度比的抑制程度。半抑制濃度(IC50)計算是用GraphPad Prism軟體的sigmoidal dose-response功能擬合得到。 The process is briefly described as follows. Ramos cells were first serum-starved for 2 hours in RPMI1640 medium containing 0.5% fetal bovine serum. Thereafter, the cells were mixed with different concentrations of compounds and incubated for 1 hour in a CO 2 incubator. After the incubation, the cells were stimulated with 1 mmol of sodium vanadate or sodium pervanadate for 20 minutes. Then, the cells were collected by centrifugation, and the cells were incubated with a 1-fold concentration of cell lysate (a 4-fold stock solution provided in the kit) at room temperature (25 degrees Celsius) for 10 minutes to lyse the cells. During the incubation, the antibody mixture was prepared and the BTK-d2 antibody and pBTK-K antibody were diluted in assay buffer (provided in the kit). In the OptiPlate-384 assay plate (6005620, PerkinElmer), 2 microliters of antibody mix was added to each well. Then, 18 μl of the cell lysate after lysing the cells was transferred to the microwells previously added to the antibody mixture. Gently mix and centrifuge briefly, block with a sealing membrane and incubate at room temperature for 18 hours at room temperature. After the completion of the incubation, emission fluorescence at two wavelengths of 665 nm and 620 nm was measured in a plate reader (PHERAstar FS, BMG). The activity calculation of the compound is based on the degree of inhibition of the signal intensity ratio at 665 nm and 620 nm. The semi-inhibitory concentration (IC50) calculation was obtained by fitting the sigmoidal dose-response function of the GraphPad Prism software.
本發明公開了被檢測的具有代表性的化合物，發現它們能夠抑制Btk和Btk在223位酪氨酸的自身磷酸化，IC50值的範圍從次納摩爾到10微摩爾。 The present invention discloses representative compounds that were detected and found to inhibit autophosphorylation of Btk and Btk at position 223 with IC50 values ranging from sub-nanomol to 10 micromolar.
n.d.：沒數據 N.d.: no data
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