TW202116753A - Cinnolines as inhibitors of hpk 1 - Google Patents
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Abstract
Description
本發明涉及調控HPK1功能、並應用於治療HPK1介導的疾病和病症如癌症的化合物。The present invention relates to compounds that regulate the function of HPK1 and are applied to treat HPK1-mediated diseases and disorders such as cancer.
根據腫瘤的位置和級別以及疾病所處的階段，腫瘤學家目前使用的主要癌症治療方式是手術、化療和放療。理想的治療目標是能夠完全消除腫瘤而不損害身體其他部位。不幸的是，手術切除、放射治療和化療並不特定地針對患病細胞，還會同時損害健康細胞。靶向治療基於抗原的腫瘤特異性表達或某些類型癌症中特異性蛋白的不適當過表達，但是腫瘤細胞容易發生突變, 並對這些治療藥物產生耐藥性。Depending on the location and grade of the tumor and the stage of the disease, the main cancer treatments currently used by oncologists are surgery, chemotherapy, and radiotherapy. The ideal treatment goal is to be able to completely eliminate the tumor without damaging other parts of the body. Unfortunately, surgical resection, radiation therapy, and chemotherapy do not specifically target diseased cells and can also damage healthy cells at the same time. Targeted therapy is based on tumor-specific expression of antigens or inappropriate overexpression of specific proteins in certain types of cancers, but tumor cells are prone to mutations and develop resistance to these therapeutic drugs.
隨著對癌症生物過程的理解加深，癌症免疫療法成為一種有效的治療方法，該方法引入患者自身的免疫系統對抗癌症。旨在引發或放大免疫反應的免疫療法被歸類為啟動免疫療法，而減少或抑制免疫反應的免疫療法被歸類為抑制免疫療法，其中之一是抑制負調節因數的免疫反應，這些免疫反應通常起到維持外周耐受，允許腫瘤抗原被識別為非自我實體的功能。With the deepening of understanding of the biological process of cancer, cancer immunotherapy has become an effective treatment method, which introduces the patient's own immune system to fight cancer. Immunotherapy designed to induce or amplify an immune response is classified as initiating immunotherapy, while immunotherapy that reduces or suppresses immune response is classified as suppressive immunotherapy. One of them is to suppress immune responses with negative regulatory factors. These immune responses It usually functions to maintain peripheral tolerance and allow tumor antigens to be recognized as non-self entities.
造血祖細胞激酶1（HPK1，也稱為MAP4K1）是一種造血區室活化（例如T細胞，B細胞和樹突細胞）的一種負調節因數，抑制T細胞受體（TCR）信號傳導和T細胞增殖。HPK1主要由造血細胞包括早期祖細胞表達，HPK1活化這些細胞的JNK / SAPK途徑。在TCR活化時，細胞溶質中的HPK1募集到質膜上，其中它的殘基Tyr381、Ser171和Thr165被Lck / Zap70磷酸化 （Shui JRetal .Nat. Immunol. 2007, 8, 84-91）。在T細胞中，HPK1被認為通過降低信號傳導微團簇的持久性來負調節T細胞活化（Di Bartolo et al. JEM 2007, 204: 681-691），這導致14-3-3蛋白質的募集，並和磷酸化的SLP76和Gads結合，從含LAT的微團簇中釋放SLP76-Gads-14-3-3複合物（Lasserreet al. J Cell Biol 2011, 195: 839-853）。除了TCR信號傳導之外， HPK1還以PKA依賴性方式通過前列腺素E2 （PGE2） 受體來負調節T細胞信號傳導（Sawasdikosol, S. et al, Blood 2003, 101, 3687-3689）。進一步的研究表明HPK1在活化誘導細胞死亡（AICD）和白細胞功能相關抗原-1（LFA-1）整合素啟動T細胞的調控中發揮作用（Brenner, D, et al, EMBO J. 2005, 24, 4279-4290; Patzak, I.M, et al, Eur. J. Immunol. 2010, 40, 3220-3225.）。Hematopoietic progenitor cell kinase 1 (HPK1, also known as MAP4K1) is a negative regulatory factor for activation of hematopoietic compartments (such as T cells, B cells, and dendritic cells), inhibiting T cell receptor (TCR) signaling and T cell proliferation . HPK1 is mainly expressed by hematopoietic cells including early progenitor cells, and HPK1 activates the JNK/SAPK pathway of these cells. When TCR is activated, HPK1 in the cytosol is recruited to the plasma membrane, where its residues Tyr381, Ser171, and Thr165 are phosphorylated by Lck/Zap70 (Shui JR etal . Nat. Immunol. 2007 , 8, 84-91). In T cells, HPK1 is thought to negatively regulate T cell activation by reducing the persistence of signaling microclusters (Di Bartolo et al. JEM 2007, 204: 681-691), which leads to the recruitment of 14-3-3 protein , And combine with phosphorylated SLP76 and Gads to release the SLP76-Gads-14-3-3 complex from the LAT-containing microclusters (Lasserreet al. J Cell Biol 2011, 195: 839-853). In addition to TCR signaling, HPK1 also negatively regulates T cell signaling through the prostaglandin E2 (PGE2) receptor in a PKA-dependent manner (Sawasdikosol, S. et al, Blood 2003, 101, 3687-3689). Further studies have shown that HPK1 plays a role in activation-induced cell death (AICD) and leukocyte function-associated antigen-1 (LFA-1) integrins to initiate T cell regulation (Brenner, D, et al, EMBO J. 2005, 24, 4279-4290; Patzak, IM, et al, Eur. J. Immunol. 2010, 40, 3220-3225.).
儘管眾多周知，噌啉具有生物學活性和治療各種疾病的潛力（Y. N. Yu, et al, Bioorg. Med. Chem. 2003, 11, 1475; A. L. Ruchelma et al, Bioorg. Med. Chem. 2004, 12, 795），由於合成的困難，缺乏用於更複雜的噌啉衍生物可行的合成路徑，使得該類化合物沒有得到廣泛的應用及用於治療用途和藥物特性的評估。Although many well-known, cinnoline has biological activity and the potential to treat various diseases (YN Yu, et al, Bioorg. Med. Chem. 2003, 11, 1475; AL Ruchelma et al, Bioorg. Med. Chem. 2004, 12, 795). Due to the difficulty of synthesis, the lack of a feasible synthetic route for more complex cinnoline derivatives has made such compounds not widely used and used in the evaluation of therapeutic uses and drug properties.
本公開描述了HPK1拮抗劑。本公開提供了具有式（I）或（Ia）所示結構的化合物，及其藥學上可接受的鹽、立體異構體、互變異構體、溶劑化物和前藥。還提供了製備式（I）或（Ia）化合物的方法。The present disclosure describes HPK1 antagonists. The present disclosure provides compounds having the structure represented by formula (I) or (Ia), and pharmaceutically acceptable salts, stereoisomers, tautomers, solvates and prodrugs thereof. A method for preparing the compound of formula (I) or (Ia) is also provided.
本公開還提供了化合物用於抑制HPK1激酶活性，增強免疫應答和治療HPK1依賴性失調的用途。相應地，還提供了包含式（I）或（Ia）化合物或其藥學上可接受的鹽，溶劑化物和前藥的藥物組合物。提供了利用有效量的式（I）或（Ia）化合物或其藥學上可接受的鹽、溶劑化物和前藥抑制HPK1的方法。提供了治療HPK1依賴性失調的方法，包括向有需要的受試者施用式（I）或（Ia）化合物或其藥物組合物。提供了式（I）或（Ia）化合物或其藥學上可接受的鹽、溶劑化物或前藥作為藥劑或者藥物的用途。該藥劑或者藥物能夠抑制HPK1激酶活性，治療HPK1依賴性失調。The present disclosure also provides the use of compounds for inhibiting HPK1 kinase activity, enhancing immune response and treating HPK1-dependent disorders. Correspondingly, a pharmaceutical composition comprising a compound of formula (I) or (Ia) or a pharmaceutically acceptable salt, solvate and prodrug thereof is also provided. A method for inhibiting HPK1 using an effective amount of a compound of formula (I) or (Ia) or a pharmaceutically acceptable salt, solvate and prodrug thereof is provided. A method for treating HPK1-dependent disorders is provided, which includes administering a compound of formula (I) or (Ia) or a pharmaceutical composition thereof to a subject in need. The use of a compound of formula (I) or (Ia) or a pharmaceutically acceptable salt, solvate or prodrug thereof as a medicament or drug is provided. The medicament or medicine can inhibit the activity of HPK1 kinase and treat HPK1-dependent disorders.
本公開提供了作為HPK1（造血祖細胞激酶1）抑制劑或調節劑的式（I）或（Ia）化合物和其藥物組合物。本公開還提供了化合物和組合物在治療由HPK1介導的疾病和失調中的用途。該治療應用能夠增強患者的免疫應答，例如以單一藥劑的形式或與其他癌症治療物組合來治療癌症患者。The present disclosure provides compounds of formula (I) or (Ia) and pharmaceutical compositions thereof as inhibitors or modulators of HPK1 (hematopoietic progenitor cell kinase 1). The present disclosure also provides the use of the compounds and compositions in the treatment of diseases and disorders mediated by HPK1. This therapeutic application can enhance the patient's immune response, for example, in the form of a single agent or in combination with other cancer treatments to treat cancer patients.
化合物Compound
本公開提供了式（I）或（Ia）化合物，或其藥學上可接受的鹽、立體異構體、互變異構體、溶劑化物和前藥。在一個方面，提供了式（I）化合物，
在另一個方面，提供了式（Ia）化合物，
在一些實施方式中，提供了式（I）或式（Ia）化合物，其中R4 為A-C(O)-；A選自由C0-6烷基、C3-7環烷基、C5-9雜芳基、C4-9雜環基、-NHR或者-OR組成的組；其中A的所述C0-6烷基、C3-7環烷基、C5-9雜芳基和C4-9雜環基任選地獨立地被一個、兩個、三個、四個或者五個取代基取代；R獨立地選自由C0-6烷基、C2-6烯基、C3-7環烷基、C6-10芳基、C5-9雜芳基和C4-9雜環基組成的組；其中R的所述C0-6烷基、C2-6烯基、C3-7環烷基、C6-10芳基、C5-9雜芳基和C4-9雜環基任選地獨立地被一個、兩個、三個、四個或者五個取代基取代。In some embodiments, a compound of formula (I) or formula (Ia) is provided, wherein R 4 is AC(O)-; A is selected from C0-6 alkyl, C3-7 cycloalkyl, C5-9 heteroaryl Group, C4-9 heterocyclic group, -NHR or -OR; wherein the C0-6 alkyl group, C3-7 cycloalkyl group, C5-9 heteroaryl group and C4-9 heterocyclic group of A are any Optionally independently substituted by one, two, three, four or five substituents; R is independently selected from C0-6 alkyl, C2-6 alkenyl, C3-7 cycloalkyl, C6-10 aryl Group, C5-9 heteroaryl group and C4-9 heterocyclic group; wherein the C0-6 alkyl group, C2-6 alkenyl group, C3-7 cycloalkyl group, C6-10 aryl group, C5 The -9 heteroaryl group and the C4-9 heterocyclic group are optionally substituted independently with one, two, three, four, or five substituents.
在一些實施方式中，提供了式（I）或式（Ia）化合物，其中R4 選自下列組成的組： C3-7環烷基， C5-9雜芳基， 獨立地被一個、兩個、三個、四個或者五個取代基取代的C4-9雜環基，所述取代基選自由具有1-4個選自O、S和N的雜原子的5-10元雜環基組成的組；和 與環稠合的5-10元雜環基，所述環選自由5-或6-元雜芳基、5-10元雜環基、C6-10芳基和C3-7環烷基組成的組，其中所述5-10元雜環基和稠合的環任選地被一個、兩個、三個、四個或者五個取代基取代。In some embodiments, a compound of formula (I) or formula (Ia) is provided, wherein R 4 is selected from the group consisting of: C3-7 cycloalkyl, C5-9 heteroaryl, independently of one or two , C4-9 heterocyclic group substituted by three, four or five substituents, the substituents are selected from 5-10 membered heterocyclic groups having 1-4 heteroatoms selected from O, S and N And a 5-10 membered heterocyclic group fused to a ring, the ring selected from the group consisting of 5- or 6-membered heteroaryl, 5-10 membered heterocyclic, C6-10 aryl and C3-7 ring The group consisting of an alkyl group, wherein the 5-10 membered heterocyclic group and the fused ring are optionally substituted with one, two, three, four or five substituents.
在一些實施方式中，提供了式（I）或式（Ia）化合物，其中R2 為具有1個、2個、3個或4個選自O、S和N的雜原子的5或6元雜芳基，並且任選被一個、兩個、三個、四個或五個取代基取代（例如，所述雜芳基為吡啶基、咪唑基、噻唑基、吡唑基或惡唑基）。In some embodiments, a compound of formula (I) or formula (Ia) is provided, wherein R 2 is a 5- or 6-membered compound having 1, 2, 3, or 4 heteroatoms selected from O, S, and N Heteroaryl, optionally substituted with one, two, three, four or five substituents (for example, the heteroaryl is pyridyl, imidazolyl, thiazolyl, pyrazolyl or oxazolyl) .
在一些實施方式中，提供了式（I）或式（Ia）化合物，其中R2 為具有1個、2個、3個或4個選自O、S和N的雜原子的5、6或7元雜環基，並且任選地被一個、兩個、三個、四個或五個取代基取代，其中雜環基為飽和或者部分不飽和的（例如，上述雜環基為惡唑烷基）。In some embodiments, a compound of formula (I) or formula (Ia) is provided, wherein R 2 is 5, 6 or 5, 6 or 4 with 1, 2, 3 or 4 heteroatoms selected from O, S and N A 7-membered heterocyclic group is optionally substituted with one, two, three, four or five substituents, wherein the heterocyclic group is saturated or partially unsaturated (for example, the above-mentioned heterocyclic group is oxazolidine base).
在一些實施方式中，提供了式（I）或式（Ia）化合物，其中R2 為任選地被一個、兩個、三個、四個或五個取代基取代的C6-10芳基（例如，上述芳基為苯基）。In some embodiments, there is provided a compound of formula (I) or formula (Ia), wherein R 2 is a C6-10 aryl group optionally substituted with one, two, three, four, or five substituents ( For example, the aforementioned aryl group is phenyl).
在一些實施方式中，提供了式（I）或式（Ia）化合物，其中R2 為C6-10芳基，或具有1個、2個、3個或4個選自O、S和N的雜原子的5或6元雜芳基；其中所述C6-10芳基或所述5或6元雜芳基與具有1個、2個、3個或4個選自O、S和N的雜原子的飽和或者部分不飽和的5、6或7元雜環基稠合；其中所述芳基、雜芳基和雜環基任選地被一個、兩個、三個或者四個取代基取代（例如，上述芳基或雜芳基與上述雜環基稠合，形成2,3-二氫-1 H-吡啶並[2,3-b][1,4]惡嗪基、 2-氫-3H-苯並[d]惡唑基或者2,3-二氫-1H-吡啶並[2,3-b][1,4]噻嗪基）。In some embodiments, a compound of formula (I) or formula (Ia) is provided, wherein R 2 is a C6-10 aryl group, or has 1, 2, 3, or 4 selected from O, S, and N Heteroatomic 5- or 6-membered heteroaryl group; wherein the C6-10 aryl group or the 5- or 6-membered heteroaryl group has 1, 2, 3 or 4 selected from O, S and N Heteroatomic saturated or partially unsaturated 5, 6 or 7-membered heterocyclic groups are fused; wherein the aryl, heteroaryl and heterocyclic groups are optionally substituted by one, two, three or four substituents Substitution (for example, the above-mentioned aryl or heteroaryl group is fused with the above-mentioned heterocyclic group to form 2,3-dihydro-1 H-pyrido[2,3-b][1,4]oxazinyl, 2- Hydrogen-3H-benzo[d]oxazolyl or 2,3-dihydro-1H-pyrido[2,3-b][1,4]thiazinyl).
在一些實施方式中，提供了式（I）或式（Ia）化合物，其中R2 上的取代基或者有關R2 時所提及的取代基，在可行的條件下，獨立地選自由C1-6烷基、氨基、C1-6烷氧基、氧代、(C1-6烷基)2 P(=O)-、鹵素、氰基和C1-6鹵代烷基組成的組。In some embodiments, provided compounds of formula (I) or Formula (Ia) compounds in which the substituents on R 2, or about 2 R substituents mentioned under practical conditions, are independently selected from C1- 6A group consisting of alkyl, amino, C1-6 alkoxy, oxo, (C1-6 alkyl) 2 P(=O)-, halogen, cyano and C1-6 haloalkyl.
在一些實施方式中，提供了式（I）化合物，其中R3 選自由H或Cl組成的組。In some embodiments, there is provided a compound of formula (I), wherein R 3 is selected from the group consisting of H or Cl.
在一些實施方式中，提供了式（I）或式（Ia）化合物，其中A為C3、C4、C5或C6環烷基，或具有1個、2個或3個選自O、S和N的雜原子的飽和或部分不飽和的4、5或6元雜環基；其中所述環烷基或者所述雜環基任選地被一個、兩個、三個、四個或者五個取代基取代（例如，所述雜環基為四氫呋喃基、氮雜環丁烷基或四氫吡喃基）。In some embodiments, a compound of formula (I) or formula (Ia) is provided, wherein A is C3, C4, C5 or C6 cycloalkyl, or has 1, 2, or 3 selected from O, S and N A heteroatomic saturated or partially unsaturated 4, 5 or 6-membered heterocyclic group; wherein the cycloalkyl group or the heterocyclic group is optionally substituted by one, two, three, four or five Group substitution (for example, the heterocyclic group is tetrahydrofuranyl, azetidinyl, or tetrahydropyranyl).
在一些實施方式中，提供了式（I）或式（Ia）化合物，其中A為-OR，其中R為具有1個、2個或3個選自O、S和N的雜原子的飽和或者部分不飽和的4、5或6元雜環基，其中所述雜環基任選地被一個、兩個、三個、四個或五個取代基取代（例如，所述雜環基為四氫吡喃基）。In some embodiments, a compound of formula (I) or formula (Ia) is provided, wherein A is -OR, wherein R is saturated with 1, 2, or 3 heteroatoms selected from O, S, and N, or Partially unsaturated 4-, 5- or 6-membered heterocyclic group, wherein the heterocyclic group is optionally substituted with one, two, three, four or five substituents (for example, the heterocyclic group is four Hydrogen pyranyl).
在一些實施方式中，提供了式（I）或式（Ia）化合物，其中D為具有1個、2個、3個或4個選自O、S和N的雜原子的5或6元雜芳基，其中所述雜芳基任選地被一個、兩個、三個、四個或五個取代基取代（例如，上述雜芳基為吡唑基、吡啶基、吡嗪基或噠嗪基）。In some embodiments, a compound of formula (I) or formula (Ia) is provided, wherein D is a 5- or 6-membered heteroatom having 1, 2, 3, or 4 heteroatoms selected from O, S, and N. An aryl group, wherein the heteroaryl group is optionally substituted with one, two, three, four or five substituents (for example, the above-mentioned heteroaryl group is pyrazolyl, pyridinyl, pyrazinyl or pyridazine base).
在一些實施方式中，提供了式（I）或式（Ia）化合物，其中D為具有1個、2個、3個或4個選自O、S和N的雜原子的、且與5、6或7元雜環基稠合的5或6元雜芳基；所述5、6或7元雜環基具有1個、2個或3個選自O、S和N的雜原子、且為飽和或者部分不飽和的；其中所述雜芳基和所述雜環基任選地被一個、兩個、三個、四個或五個取代基所取代（例如，上述雜芳基和上述雜環基稠合形成5,6,7,8-四氫-4H-吡唑並[1,5-d][1,4]二氮雜卓基、5,6,7,8-四氫-1,6-萘啶或者4,5-二氫-7H-吡唑並[5,1-d][1,2,5]噻二嗪基）。In some embodiments, a compound of formula (I) or formula (Ia) is provided, wherein D is one having 1, 2, 3, or 4 heteroatoms selected from O, S, and N, and is combined with 5, A 5- or 6-membered heteroaryl group fused with a 6- or 7-membered heterocyclic group; the 5-, 6- or 7-membered heterocyclic group has 1, 2, or 3 heteroatoms selected from O, S and N, and Is saturated or partially unsaturated; wherein the heteroaryl group and the heterocyclic group are optionally substituted with one, two, three, four or five substituents (for example, the above-mentioned heteroaryl group and the above-mentioned Heterocyclic groups are fused to form 5,6,7,8-tetrahydro-4H-pyrazolo[1,5-d][1,4]diazepine, 5,6,7,8-tetrahydro -1,6-naphthyridine or 4,5-dihydro-7H-pyrazolo[5,1-d][1,2,5]thiadiazinyl).
在一些實施方式中，提供了式（I）或式（Ia）化合物，其中A或D上的取代基或者有關於A或D的取代基，在可行的條件下，獨立地選自由C1-C6烷基，氰基，氧代，具有1個、2個、3個或者4個選自O、S和N的雜原子的5或6元雜芳基，C1-6烷基-C(=O)- ，C1-6烷基-S(=O)2 -，具有1個、2個或3個選自O、S和N 的雜原子的飽和或者部分不飽和的5、6或7元雜環基，(具有1個、2個或3個選自O、S和N的雜原子的飽和或者部分不飽和的5、6或7元雜環基)-C(=O)-，C1-6亞烷基和鹵素（例如，上述雜芳基為吡唑基）（例如，上述雜環基為嗎啉基、呱嗪基、硫嗎啉基、吡咯烷基、呱啶基或氮雜環丁基）；In some embodiments, a compound of formula (I) or formula (Ia) is provided, wherein the substituent on A or D or the substituent on A or D is independently selected from C1-C6 when feasible Alkyl, cyano, oxo, 5 or 6-membered heteroaryl with 1, 2, 3 or 4 heteroatoms selected from O, S and N, C1-6 alkyl-C(=O )-, C1-6 alkyl-S(=O) 2 -, saturated or partially unsaturated 5, 6 or 7-membered heteroatoms with 1, 2 or 3 heteroatoms selected from O, S and N Cyclic group, (saturated or partially unsaturated 5, 6 or 7-membered heterocyclic group with 1, 2 or 3 heteroatoms selected from O, S and N) -C(=O)-, C1- 6 Alkylene and halogen (for example, the above heteroaryl group is pyrazolyl) (for example, the above heterocyclic group is morpholinyl, azizinyl, thiomorpholinyl, pyrrolidinyl, pyridinyl or nitrogen heterocycle Butyl);
其中，上述取代基在可行的條件下，進一步任選地被一個、兩個、三個、四個或五個獨立地選自下列組成的組的取代基取代：氰基，C1-6烷基，C1-6烷基-S(=O)2 -，C1-6 烷基-C(=O)-，氧代，具有1個、2個或3個選自O、S和N的雜原子的飽和或部分不飽和的5、6或7元雜環基，具有1個、2個、3個或4個選自O、S和N的雜原子的5或6元雜芳基，和 C3、C4、C5或C6環烷基（例如，上述雜環基為氧雜環丁烷基或氮雜環丁烷基）（例如，上述雜芳基為吡唑基）；和Wherein, the above-mentioned substituents are further optionally substituted with one, two, three, four or five substituents independently selected from the group consisting of: cyano, C1-6 alkyl when feasible , C1-6 alkyl-S(=O) 2 -, C1-6 alkyl-C(=O)-, oxo, with 1, 2, or 3 heteroatoms selected from O, S and N A saturated or partially unsaturated 5-, 6- or 7-membered heterocyclic group, a 5- or 6-membered heteroaryl group with 1, 2, 3 or 4 heteroatoms selected from O, S and N, and C3 , C4, C5 or C6 cycloalkyl (for example, the above heterocyclic group is oxetanyl or azetidinyl) (for example, the above heteroaryl is pyrazolyl); and
其中，上述進一步的取代基在可行的條件下，任選地被一個、兩個、三個、四個或五個獨立地選自由鹵素、C1-6烷氧基、C1-6 烷基-S(=O)2 -、羥基、C1-6烷基-C(=O)- 和C1-6 烷基組成的組所取代。Wherein, the above-mentioned further substituents are optionally selected by one, two, three, four or five independently selected from halogen, C1-6 alkoxy, C1-6 alkyl-S under feasible conditions. (=O) 2 -, hydroxy, C1-6 alkyl-C(=O)- and C1-6 alkyl substituted.
在一些實施方式中，提供了式（Ia）所示的化合物，其中：
R1
是-C-M；
M是H，鹵素，CN；
R2
選自下列基團之一及其對映體：
在一些實施方式中，提供了式（Ia）所示化合物，其中R4
選自下列及其相應的對映體或非對映體：
在一些實施方式中，提供了式（I）或式（Ia）化合物，其中如果提到C0-6烷基，其為H、C1烷基、C2烷基、C3烷基、C4烷基、C5烷基或C6烷基。In some embodiments, a compound of formula (I) or formula (Ia) is provided, wherein if C0-6 alkyl is mentioned, it is H, C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 Alkyl or C6 alkyl.
在一些實施方式中，提供了式（I）或式（Ia）化合物，其中如果提到C5-9雜芳基，其為具有1個、2個、3個或4個選自O、S和N的雜原子的5、6、7、8或9元雜芳基。In some embodiments, a compound of formula (I) or formula (Ia) is provided, wherein if a C5-9 heteroaryl group is mentioned, it has 1, 2, 3 or 4 selected from O, S and 5, 6, 7, 8 or 9-membered heteroaryl groups of N heteroatoms.
在一些實施方式中，提供了式（I）或式（Ia）化合物，其中如果提到C4-9雜環基，其為具有1個、2個、3個或4個選自O、S和N的雜原子的飽和或部分不飽和的4、5、6、7、8或9元雜環基。In some embodiments, a compound of formula (I) or formula (Ia) is provided, wherein if a C4-9 heterocyclic group is mentioned, it has 1, 2, 3 or 4 selected from O, S and A saturated or partially unsaturated 4, 5, 6, 7, 8 or 9-membered heterocyclic group of N heteroatoms.
在一些實施方式中，化合物選自表1：In some embodiments, the compound is selected from Table 1:
表1.式（I）和（Ia）的代表性化合物
[HPK1激酶][HPK1 Kinase]
造血祖細胞激酶1（HPK1，也稱為MAP4K1）作為造血區室細胞活化（例如T細胞，B細胞和樹突細胞）的負調節物的一種示例，能夠抑制T細胞受體（TCR）信號傳導和T細胞增殖。因為它涉及到被認為限制T細胞反應性的幾個重要步驟，特別是在癌症中，使得HPK1成為特別令人感興趣的靶點。Hematopoietic progenitor kinase 1 (HPK1, also known as MAP4K1) is an example of a negative regulator of hematopoietic compartment cell activation (such as T cells, B cells, and dendritic cells), which can inhibit T cell receptor (TCR) signaling and T cell proliferation. Because it involves several important steps that are thought to limit the reactivity of T cells, especially in cancer, making HPK1 a particularly interesting target.
HPK1主要由造血細胞包括早期祖細胞表達，HPK1可活化這些細胞的 JNK / SAPK途徑。在TCR活化時，細胞溶質中的HPK1募集到質膜，其中它的殘基Tyr381、Ser171和Thr165被Lck / Zap70磷酸化（Shui JRetal. Nat.Immunol.2007, 8, 84-91）。在T細胞中，HPK1被認為通過減少信號傳導微團簇的持久性來負調節T細胞活化（Di Bartolo等人(2007)JEM 204：681-691），導致14-3-3蛋白質的募集，並結合磷酸化的SLP76和Gads，使得從含LAT的微團簇中釋放SLP76-Gads-14-3-3複合物（Lasserreetal. J Cell Biol 2007, 195：839-853)。除了TCR信號傳導之外，HPK1還以PKA依賴性方式通過前列腺素E2（PGE2）受體負調節T細胞信號傳導（Sawasdikosol,S. et al, Blood 2003, 101, 3687-3689)。進一步的研究表明HPK1在啟動誘導的細胞死亡（AICD）和白細胞功能相關抗原1（LFA-1）整合素啟動T細胞的調控中發揮作用 （Brenner, D.,et al, EMBO J. 2005, 24, 4279-4290; Patzak, I. M, et al, Eur.J. Immunol. 2010 40, 3220-3225）。HPK1 is mainly expressed by hematopoietic cells including early progenitor cells, and HPK1 can activate the JNK/SAPK pathway of these cells. When TCR is activated, HPK1 in the cytosol is recruited to the plasma membrane, where its residues Tyr381, Ser171, and Thr165 are phosphorylated by Lck/Zap70 (Shui JRetal. Nat. Immunol. 2007, 8, 84-91). In T cells, HPK1 is thought to negatively regulate T cell activation by reducing the persistence of signaling microclusters (Di Bartolo et al. (2007) JEM 204:681-691), leading to the recruitment of 14-3-3 protein, And combined with phosphorylated SLP76 and Gads, so that the SLP76-Gads-14-3-3 complex is released from the LAT-containing microclusters (Lasserreetal. J Cell Biol 2007, 195: 839-853). In addition to TCR signaling, HPK1 negatively regulates T cell signaling through the prostaglandin E2 (PGE2) receptor in a PKA-dependent manner (Sawasdikosol, S. et al, Blood 2003, 101, 3687-3689). Further studies have shown that HPK1 plays a role in the initiation of induced cell death (AICD) and leukocyte function-associated antigen 1 (LFA-1) integrin to initiate the regulation of T cells (Brenner, D., et al, EMBO J. 2005, 24 , 4279-4290; Patzak, I. M, et al, Eur. J. Immunol. 2010 40, 3220-3225).
來自HPK1敲除小鼠的骨髓來源的樹突細胞（BDMC）顯示出共刺激分子（例如CD80 / CD86）的更高表達和促炎細胞因數（IL-12、TNF-α等）產量的提高，並且與野生型DC相比，在體內和體外均表現出優異的刺激T細胞增殖的能力 （Alzabin, S. et al, J Immunol, 2009, 182: 6187-94)。這些資料表明HPK1也是樹突細胞活化的一種重要的負調節物（Alzabin, S.,et al, J Immunol, 2009.182: 6187-94)。然而，HPK-1介導的DC活化的負調節信號傳導機制仍有待闡明。Bone marrow-derived dendritic cells (BDMC) from HPK1 knockout mice showed higher expression of costimulatory molecules (such as CD80/CD86) and increased production of pro-inflammatory cytokines (IL-12, TNF-α, etc.), And compared with wild-type DC, it shows excellent ability to stimulate T cell proliferation in vivo and in vitro (Alzabin, S. et al, J Immunol, 2009, 182: 6187-94). These data indicate that HPK1 is also an important negative regulator of dendritic cell activation (Alzabin, S., et al, J Immunol, 2009.182: 6187-94). However, the negative regulatory signal transduction mechanism of HPK-1 mediated DC activation remains to be elucidated.
相反，HPK1是調節性T細胞（Treg）的抑制功能的正調節劑（Sawasdikosol,S.et al, The journal of immunology, 2012.188, supplement1：163)。HPK1缺陷小鼠Foxp3 + Tregs在抑制TCR誘導的效應T細胞增殖方面存在缺陷, 並且獲得了在TCR參與後產生IL-2的能力。這些資料表明HPK1是Treg功能和外周自身耐受的一種重要的調節因數。In contrast, HPK1 is a positive regulator of the inhibitory function of regulatory T cells (Treg) (Sawasdikosol, S. et al, The journal of immunology, 2012.188, supplement 1: 163). Foxp3 + Tregs in HPK1-deficient mice have defects in inhibiting TCR-induced effector T cell proliferation, and have acquired the ability to produce IL-2 after TCR is involved. These data indicate that HPK1 is an important regulatory factor for Treg function and peripheral self-tolerance.
還提供了式（I）和式（Ia）化合物，或其藥學上可接受的鹽、前藥或溶劑化物。在某些實施方式中，本文還提供了式（I）和式（Ia）化合物的結晶形式和無定形形式，或其藥學上可接受的鹽、前藥或溶劑化物。Also provided are compounds of formula (I) and formula (Ia), or pharmaceutically acceptable salts, prodrugs or solvates thereof. In certain embodiments, the crystalline and amorphous forms of the compounds of Formula (I) and Formula (Ia), or pharmaceutically acceptable salts, prodrugs or solvates thereof are also provided herein.
“藥學上可接受的鹽”是指通過常規方法製備的鹽，並且是本領域技術人員熟知的。所述“藥學上可接受的鹽”包括無機酸和有機酸的鹼性鹽（Berge et al., J.Pharm. Sci. 1977, 66:1)。"Pharmaceutically acceptable salt" refers to a salt prepared by a conventional method and is well known to those skilled in the art. The "pharmaceutically acceptable salt" includes basic salts of inorganic acids and organic acids (Berge et al., J. Pharm. Sci. 1977, 66:1).
通過在溶劑中處理化合物形成“溶劑化物”。還提供了式（I）和式（Ia）化合物的鹽的溶劑化物。在用水處理化合物的情況下，溶劑化物為水合物。還提供了式（I）和式（Ia）化合物的水合物。A "solvate" is formed by treating the compound in a solvent. Solvates of the salts of the compounds of formula (I) and formula (Ia) are also provided. In the case of treating the compound with water, the solvate is a hydrate. Also provided are hydrates of the compounds of formula (I) and formula (Ia).
“前藥”包括例如根據前藥的代謝過程，當其施用於受試者時，能夠轉化為式（I）和式（Ia）化合物的任何化合物。"Prodrug" includes, for example, any compound that can be converted into a compound of formula (I) and formula (Ia) when it is administered to a subject according to the metabolic process of the prodrug.
化合物的治療用途Therapeutic use of the compound
式（I）和式（Ia）化合物或其藥學上可接受的鹽、前藥或溶劑化物可用於治療HPK1介導的疾病或失調。在一個實施方案中，提供了使用式（I）和式（Ia）所示化合物或其藥學上可接受的鹽，前藥或溶劑化物抑制HPK1活性的方法。The compounds of formula (I) and formula (Ia) or pharmaceutically acceptable salts, prodrugs or solvates thereof can be used to treat HPK1-mediated diseases or disorders. In one embodiment, there is provided a method for inhibiting the activity of HPK1 using a compound represented by formula (I) and formula (Ia) or a pharmaceutically acceptable salt, prodrug or solvate thereof.
如本文所用，關於失調的“治療（treat）”或“治療（treating）”意指改善或預防失調或者改善或預防失調的一種或多種生物學表現，以阻斷引起或導致失調的生物通路中的一個或多個點，以減輕與該失調相關的一種或多種症狀或作用。如上所述，失調的“治療”包括預防失調，並且“預防”應理解為為顯著降低失調或其生物學表現的可能性或嚴重性，或延遲這種失調的發作或其生物學表現，而預防性地施用某藥物。As used herein, "treat" or "treating" in relation to a disorder means to improve or prevent the disorder or to improve or prevent one or more of the biological manifestations of the disorder in order to block the biological pathways that cause or cause the disorder One or more points in order to alleviate one or more symptoms or effects associated with the disorder. As mentioned above, the "treatment" of a disorder includes the prevention of the disorder, and "prevention" should be understood to mean significantly reducing the likelihood or severity of the disorder or its biological manifestations, or delaying the onset of such disorder or its biological manifestations, and To administer a drug prophylactically.
“受試者”是指人（包括成人和兒童）或其他動物。在一個實施方案中，“患者”是指人。"Subject" refers to humans (including adults and children) or other animals. In one embodiment, "patient" refers to a human.
如本文所用，關於式（I）和式（Ia）所示化合物或其藥學上可接受的鹽，前藥或溶劑化物的“安全有效劑量”，是其量足以治療患者的病症，但足夠低以避免嚴重副作用。某種化合物的安全有效劑量將隨所選的特定化合物 （例如考慮化合物的效力，功效和半衰期）；選擇的給藥途徑；被治療的失調；被治療的失調的嚴重程度；被治療患者的年齡、體型、體重和身體狀況；待治療患者的病史；治療的持續時間；同時治療的性質；理想的治療效果；和類似的因素而變化。As used herein, the "safe and effective dose" of the compound represented by formula (I) and formula (Ia) or a pharmaceutically acceptable salt, prodrug or solvate thereof is an amount sufficient to treat the patient's condition, but low enough To avoid serious side effects. The safe and effective dose of a certain compound will depend on the specific compound selected (for example, considering the potency, efficacy and half-life of the compound); the chosen route of administration; the disorder being treated; the severity of the disorder being treated; the age of the patient being treated , Body type, weight and physical condition; medical history of the patient to be treated; duration of treatment; nature of simultaneous treatment; ideal treatment effect; and similar factors.
“HPK1活性的抑制”或變體是指相較於在不存在式（I）和式（Ia）化合物或其藥學上可接受的鹽，前藥或溶劑化物時的HPK1的活性來說，在式（I）和式（Ia）化合物或其藥學上可接受的鹽，前藥或溶劑化物存在時，HPK1活性的降低作為一種直接或間接反應。"Inhibition of HPK1 activity" or variant refers to the activity of HPK1 in the absence of compounds of formula (I) and formula (Ia) or their pharmaceutically acceptable salts, prodrugs or solvates. When the compounds of formula (I) and formula (Ia) or their pharmaceutically acceptable salts, prodrugs or solvates exist, the reduction of HPK1 activity is a direct or indirect reaction.
作為酶活性（或其他生物活性）抑制劑的化合物的效力可以通過檢測每種化合物抑制活性達到預定程度時的濃度，然後比較結果來確定。抑制劑的“IC 50”或“IC 90”可以通過抑制生化實驗中50％或90％活性時的濃度來確定，這可以使用本領域已知的常規技術完成，包括下面實施例中描述的技術。The effectiveness of compounds as inhibitors of enzyme activity (or other biological activities) can be determined by detecting the concentration of each compound at a predetermined level of inhibitory activity, and then comparing the results. The "IC 50" or "IC 90" of the inhibitor can be determined by the concentration at which 50% or 90% of the activity is inhibited in a biochemical experiment. This can be done using conventional techniques known in the art, including the techniques described in the following examples .
HPK1主要在造血區室細胞（例如，T細胞，B細胞和樹突細胞）中表達，並且通過T細胞受體銜接蛋白SLP-76的磷酸化和活化，表明其為T細胞中信號轉導的一種負調節物（Di Bartoloet al, J.Exp.Med. 2007, 204: 681）。基於體外和體內研究，HPK1被鑒定為癌症免疫療法的新靶標（Sawasdikosolet al, Immunol Res.2012, 54: 262-5）。HPK1 is mainly expressed in hematopoietic compartment cells (for example, T cells, B cells and dendritic cells), and through phosphorylation and activation of the T cell receptor adaptor protein SLP-76, indicating that it is a kind of signal transduction in T cells Negative regulators (Di Bartolo et al, J. Exp. Med. 2007, 204: 681). Based on in vitro and in vivo studies, HPK1 has been identified as a new target for cancer immunotherapy (Sawasdikosolet al, Immunol Res. 2012, 54: 262-5).
在一個方面，本發明因此提供了一種治療由不適當的HPK1活性介導的失調的方法，包括給予安全有效劑量的式（I）和式（Ia）所示化合物，或其藥學上可接受的鹽，前藥或溶劑化物。In one aspect, the present invention therefore provides a method for treating disorders mediated by inappropriate HPK1 activity, comprising administering a safe and effective dose of a compound represented by formula (I) and formula (Ia), or a pharmaceutically acceptable Salt, prodrug or solvate.
在一個實施方式中，本文描述的化合物可用於治療由不適當的HPK1活性介導的癌症。在某些實施方案中，該疾病是惡性血液病。在某些實施方案中，所述疾病是淋巴瘤，例如伯基特淋巴瘤、彌漫性大B細胞淋巴瘤（DLBCL）和套細胞淋巴瘤（MCL）、濾泡性淋巴瘤、淋巴漿細胞性淋巴瘤、瓦爾登斯特倫巨球蛋白血症（Waldenstrom macroglobulinemia）和邊緣區淋巴瘤。在一個實施方案中，所述病症是多發性骨髓瘤或白血病，例如急性淋巴細胞白血病（ALL）、急性髓性白血病（AML）、慢性淋巴細胞白血病（CLL）、小淋巴細胞性淋巴瘤（SLL）、骨髓增生異常綜合征（MDS）、骨髓增生性疾病（MPD）、慢性粒細胞白血病（CML）。In one embodiment, the compounds described herein can be used to treat cancers mediated by inappropriate HPK1 activity. In certain embodiments, the disease is hematological malignancy. In certain embodiments, the disease is lymphoma, such as Burkitt’s lymphoma, diffuse large B-cell lymphoma (DLBCL) and mantle cell lymphoma (MCL), follicular lymphoma, lymphoplasmacytic Lymphoma, Waldenstrom macroglobulinemia and marginal zone lymphoma. In one embodiment, the condition is multiple myeloma or leukemia, such as acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL) ), myelodysplastic syndrome (MDS), myelodysplastic disease (MPD), chronic myelogenous leukemia (CML).
在其他實施方案中，該疾病是實體瘤。在具體的實施方案中，所述適應症用於治療具有異常HPK1表達的實體瘤，例如胰腺導管腺癌(PDAC)和肝細胞癌(HCC)、胃腸癌、前列腺癌、卵巢癌、成神經管細胞瘤和乳腺癌。在一些實施方案中，所述化合物可單獨或與其他抗癌療法聯合，用於治療前列腺癌、膀胱癌、結腸直腸癌、腎癌、肝細胞癌、肺癌、卵巢癌、宮頸癌、頭頸癌、黑色素瘤、神經內分泌癌、腦腫瘤、骨癌或軟組織肉瘤。In other embodiments, the disease is a solid tumor. In a specific embodiment, the indication is used to treat solid tumors with abnormal HPK1 expression, such as pancreatic ductal adenocarcinoma (PDAC) and hepatocellular carcinoma (HCC), gastrointestinal cancer, prostate cancer, ovarian cancer, neural tube Cell tumor and breast cancer. In some embodiments, the compound may be used alone or in combination with other anti-cancer therapies for the treatment of prostate cancer, bladder cancer, colorectal cancer, kidney cancer, hepatocellular carcinoma, lung cancer, ovarian cancer, cervical cancer, head and neck cancer, Melanoma, neuroendocrine cancer, brain tumor, bone cancer or soft tissue sarcoma.
聯合療法Combination therapy
在一個實施方式中，式（I）和式（Ia）所示化合物或其藥學上可接受的鹽，前藥或溶劑化物可以與一種或多種另外的治療劑組合使用以治療癌症或炎性病症。一種或多種另外的治療劑可以是化學治療劑、放射療法、靶向療法、免疫治療劑，或者作為小分子或具備生物學性質的任何目前的最佳護理治療。In one embodiment, the compound represented by formula (I) and formula (Ia) or a pharmaceutically acceptable salt, prodrug or solvate thereof can be used in combination with one or more additional therapeutic agents to treat cancer or inflammatory disorders . The one or more additional therapeutic agents may be chemotherapeutic agents, radiotherapy, targeted therapy, immunotherapeutic agents, or any current best care treatment as a small molecule or with biological properties.
靶向治療包括但不限於細胞週期蛋白依賴性激酶（CDK），例如CDK1、CDK2、CDK4/6、CDK7和CDK9，Janus激酶（JAK）如JAK1、JAK2和/或JAK3，脾酪氨酸激酶 （SYK），Bruton酪氨酸激酶（BTK），絲裂原活化蛋白激酶（MEK）如MEK1和MEK2，含溴結構域蛋白（BRD）如BRD4，異檸檬酸脫氫酶（IDH）如IDH1，組蛋白去乙醯化酶（HDAC），PI3K激酶或其任意組合的的抑制劑。Targeted therapies include but are not limited to cyclin-dependent kinases (CDK), such as CDK1, CDK2, CDK4/6, CDK7 and CDK9, Janus kinases (JAK) such as JAK1, JAK2 and/or JAK3, and spleen tyrosine kinases ( SYK), Bruton tyrosine kinase (BTK), mitogen-activated protein kinase (MEK) such as MEK1 and MEK2, bromodomain-containing proteins (BRD) such as BRD4, isocitrate dehydrogenase (IDH) such as IDH1, group Inhibitors of protein deacetylase (HDAC), PI3K kinase or any combination thereof.
化學治療劑可以通過其作用機理分類為：烷化劑、抗代謝物、抗微管劑、拓撲異構酶抑制劑和細胞毒性劑。式（I）和式（Ia）化合物，或其藥學上可接受的鹽、前藥或溶劑化物可以與化學治療劑組合使用，以敏化和改善某些化學治療劑治療血液或實體瘤的功效。Chemotherapeutic agents can be classified into alkylating agents, antimetabolites, anti-microtubule agents, topoisomerase inhibitors, and cytotoxic agents by their mechanism of action. The compounds of formula (I) and formula (Ia), or pharmaceutically acceptable salts, prodrugs or solvates thereof can be used in combination with chemotherapeutic agents to sensitize and improve the efficacy of certain chemotherapeutic agents in the treatment of blood or solid tumors .
免疫治療劑包括但不限於適用于治療患者的治療性抗體、小分子和疫苗；例如IDO1和TDO2抑制劑，A2A受體抑制劑，精氨酸酶抑制劑，toll樣受體激動劑，趨化因數調節劑（包括CCR和CXCR家族），檢查點阻斷抗體如調節PD-1、PD-L1、CTLA-4、OX40-OX40配體、LAG3、TIM3的抗體，或其任意組合。Immunotherapeutics include, but are not limited to, therapeutic antibodies, small molecules, and vaccines suitable for treating patients; for example, IDO1 and TDO2 inhibitors, A2A receptor inhibitors, arginase inhibitors, toll-like receptor agonists, chemotaxis Factor modifiers (including CCR and CXCR families), checkpoint blocking antibodies such as antibodies that modulate PD-1, PD-L1, CTLA-4, OX40-OX40 ligand, LAG3, TIM3, or any combination thereof.
放射療法是控制或殺死惡性細胞的癌症治療的一部分，並且由於其控制細胞生長的能力而通常應用於癌性腫瘤。式（I）和式（Ia）化合物或其藥學上可接受的鹽、前藥或溶劑化物可與放射療法聯合應用，以改善放射療法治療血液或實體瘤的效果，或者和手術、化學療法、免疫療法以及四種手段聯合應用。Radiotherapy is part of cancer treatment that controls or kills malignant cells, and is often applied to cancerous tumors due to its ability to control cell growth. The compounds of formula (I) and formula (Ia) or their pharmaceutically acceptable salts, prodrugs or solvates can be used in combination with radiotherapy to improve the effect of radiotherapy in the treatment of blood or solid tumors, or with surgery, chemotherapy, Immunotherapy and the combined application of four methods.
在某些實施方式中，式（I）和式（Ia）化合物或其藥學上可接受的鹽、前藥或溶劑化物可以與一種或多種另外的治療劑組合使用，以治療用至少一種化療治療基本上難以治癒無效或化療後復發的患者。In certain embodiments, the compounds of formula (I) and formula (Ia) or pharmaceutically acceptable salts, prodrugs or solvates thereof can be used in combination with one or more additional therapeutic agents to treat at least one chemotherapeutic treatment It is basically difficult to cure patients who are ineffective or who relapse after chemotherapy.
藥物組合物Pharmaceutical composition
另一方面，本發明提供了一種藥物組合物，其包含式（I）和式（Ia）化合物或其藥學上可接受的鹽、前藥或溶劑化物以及藥學上可接受的載體或賦形劑。可配製該藥物組合物用於特定的給藥途徑，例如口服給藥、腸胃外給藥和局部給藥等。In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I) and formula (Ia) or a pharmaceutically acceptable salt, prodrug or solvate thereof, and a pharmaceutically acceptable carrier or excipient . The pharmaceutical composition can be formulated for a specific route of administration, such as oral administration, parenteral administration, and topical administration.
根據本領域已知的用於製備藥物組合物的任何方法來製備旨在 口服使用的組合物，並且可以被製備成片劑、丸劑、粉末、懸浮液、乳液、溶液、糖漿和膠囊的形式。口服組合物可以與適用於製備片劑的藥學上可接受的無毒的賦形劑混合的活性成分。片劑未經包衣或通過已知技術包衣以延遲在胃腸道中的崩解和吸收，從而在較長時間內提供持續作用。供口服使用的製劑可以呈現為硬明膠膠囊，其中活性成分與惰性固體稀釋劑混合，例如碳酸鈣、磷酸鈣或高嶺土，或呈現為軟明膠膠囊，其中活性成分與水或油介質混合，例如花生油、液體石蠟或橄欖油。The composition intended for oral use is prepared according to any method known in the art for preparing pharmaceutical compositions, and can be prepared in the form of tablets, pills, powders, suspensions, emulsions, solutions, syrups and capsules. The oral composition may be an active ingredient mixed with pharmaceutically acceptable non-toxic excipients suitable for the preparation of tablets. The tablets are uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained action over a longer period of time. Formulations for oral use can be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules, in which the active ingredient is mixed with a water or oil medium, such as peanut oil , Liquid paraffin or olive oil.
某些可注射的組合物為水性等滲溶液或懸浮液，並且由脂肪乳劑或懸浮液有利於製備栓劑。Certain injectable compositions are aqueous isotonic solutions or suspensions, and it is advantageous to prepare suppositories from fatty emulsions or suspensions.
用於透皮施用的合適組合物包括有效量的本發明的化合物和合適的載體。適於透皮遞送的載體包括可吸收的藥理學上可接受的溶劑，以幫助通過宿主的皮膚。例如，透皮設備為繃帶形式，其包括：背襯構件；容器，其含有化合物以及任選地含有載體；任選地速率控制屏障，以在長時間裡以可控和預定的速率將化合物遞送至宿主皮膚；以及將該設備固定至皮膚的裝置。Suitable compositions for transdermal administration include an effective amount of a compound of the invention and a suitable carrier. Carriers suitable for transdermal delivery include absorbable pharmacologically acceptable solvents to assist passage through the skin of the host. For example, the transdermal device is in the form of a bandage, which includes: a backing member; a container that contains a compound and optionally a carrier; optionally a rate control barrier to deliver the compound at a controlled and predetermined rate over a long period of time To the skin of the host; and a device to fix the device to the skin.
用於局部施用（例如，施用至皮膚和眼睛）的合適組合物包括水溶液、懸浮液、軟膏、乳膏、凝膠或噴霧製劑，例如通過氣溶膠等遞送。這種局部遞送系統特別適用於皮膚施用，例如用於治療皮膚癌，例如用於防曬霜、洗劑、噴霧劑等的預防性用途。Suitable compositions for topical application (for example, application to the skin and eyes) include aqueous solutions, suspensions, ointments, creams, gels, or spray formulations, such as delivery by aerosol or the like. This topical delivery system is particularly suitable for skin application, for example for the treatment of skin cancer, for example for preventive use in sunscreens, lotions, sprays and the like.
如本文所用，局部施用還可以涉及吸入或鼻內施用。它們可以使用或不使用合適的推進劑，可以方便地從乾粉吸入器以乾粉形式（或單獨，作為混合物，例如與乳糖的幹混合物，或混合組分顆粒，例如與磷脂）遞送，或從加壓容器、泵、噴霧器、霧化噴嘴或霧化器以氣溶膠噴霧呈現的形式遞送。As used herein, topical administration may also involve inhalation or intranasal administration. They can be delivered with or without a suitable propellant, and can be conveniently delivered from a dry powder inhaler in dry powder form (or alone, as a mixture, such as a dry mixture with lactose, or mixed component particles, such as with a phospholipid), or from a dry powder inhaler. The pressure vessel, pump, nebulizer, atomizing nozzle or atomizer is delivered in the form of an aerosol spray.
本發明還提供藥物組合物和劑型，其包含降低作為活性成分的本發明化合物的分解速率的一種或多種試劑。這些試劑在本文中稱為“穩定劑”，包括但不限於抗氧化劑如抗壞血酸，pH緩衝劑或鹽緩衝劑等。The present invention also provides pharmaceutical compositions and dosage forms that contain one or more agents that reduce the rate of decomposition of the compound of the present invention as an active ingredient. These agents are referred to herein as "stabilizers" and include, but are not limited to, antioxidants such as ascorbic acid, pH buffers or salt buffers, and the like.
給藥模式和劑量Mode of administration and dosage
藥物組合物可以以單劑量或多劑量給藥。基於治療目的，可以配製式（I）和（Ia）所示化合物或其藥學上可接受的鹽、前藥或溶劑化物，以提供所期望的的活性成分釋放時間表。The pharmaceutical composition can be administered in a single dose or in multiple doses. For therapeutic purposes, the compounds represented by formulas (I) and (Ia) or pharmaceutically acceptable salts, prodrugs or solvates thereof can be formulated to provide a desired release schedule of the active ingredient.
藥物組合物優選製成劑量單位的形式，含有特定量的形式為片劑、丸劑、粉劑、懸浮液、乳劑、溶液劑、糖漿劑和膠囊劑的活性成分。例如，它們可含有約0.1至1000mg，優選約0.1至500mg的活性成分。對於人或其他哺乳動物，合適的日劑量可以根據患者的狀況和其他因素而廣泛變化，但是，可以再次使用常規方法確定。日劑量可以每天給藥一至四劑量。出於治療目的，本發明的活性化合物通常與一種或多種適合於給予眼、耳或鼻的給藥途徑滴劑的佐劑組合。本發明化合物的合適局部劑量的活性成分為0.1mg至150mg，每天給藥一至四次，優選一次或兩次。對於局部給藥，活性成分可以包含占製劑重量的0.001％至10％w / w，例如，製劑重量的1％至2％，優選不超過5％w / w，更優選該製劑的0.1％至1％。The pharmaceutical composition is preferably prepared in the form of a dosage unit, containing specific amounts of active ingredients in the form of tablets, pills, powders, suspensions, emulsions, solutions, syrups and capsules. For example, they may contain about 0.1 to 1000 mg, preferably about 0.1 to 500 mg of active ingredient. For humans or other mammals, the appropriate daily dose can vary widely according to the patient's condition and other factors, but it can be determined again using conventional methods. The daily dose can be one to four doses per day. For therapeutic purposes, the active compound of the present invention is usually combined with one or more adjuvants suitable for administration to the eye, ear, or nose. A suitable topical dose of the active ingredient of the compound of the present invention is 0.1 mg to 150 mg, which is administered one to four times a day, preferably once or twice. For topical administration, the active ingredient may comprise 0.001% to 10% w/w of the preparation weight, for example, 1% to 2% of the preparation weight, preferably no more than 5% w/w, more preferably 0.1% to 10% w/w of the preparation 1%.
在一個具體實施方式中，該方法包括向受試者施用約0.1至500mg的式（I）和（Ia）化合物的初始日劑量，並通過增量來增加劑量直至達到臨床功效。可以使用約5、10、25、50或100mg的增量來增加劑量。劑量可以每天、每隔一天、每週兩次或每週一次增加。In a specific embodiment, the method includes administering to the subject an initial daily dose of a compound of formula (I) and (Ia) of about 0.1 to 500 mg, and increasing the dose by increments until clinical efficacy is achieved. The dosage can be increased in increments of about 5, 10, 25, 50, or 100 mg. The dosage can be increased daily, every other day, twice a week, or once a week.
式（I）化合物的合成Synthesis of compound of formula (I)
可以使用本文公開的方法及其常規變型來製備式（I）或（Ia）化合物，鑒於本文公開的內容，這將是顯而易見的,並且方法在本領域是公知的。除了本文的教導之外,可以使用常規和公知的合成方法。本文所述的代表性化合物的合成可以按照以下實施例中所述完成。如果可行的話，試劑可以商購，例如從Sigma Aldrich或其他化學品供應商購得。The method disclosed herein and its conventional variants can be used to prepare the compound of formula (I) or (Ia), which will be obvious in view of the content disclosed herein, and the method is well known in the art. In addition to the teachings herein, conventional and well-known synthetic methods can be used. The synthesis of the representative compounds described herein can be accomplished as described in the following examples. If feasible, reagents can be purchased commercially, for example, from Sigma Aldrich or other chemical suppliers.
綜述Summary
下面使用的試劑和溶劑可以從商業來源獲得。在Bruker 400MHZ NMR光譜儀上記錄1 H-NMR光譜。重要峰值按順序清單：多重性(s，單峰；d，雙峰；t，三重峰；q，四重峰；m，多重峰；brs，寬單峰)、以赫茲（Hz）表示的耦合常數(s)以及質子數。按照品質與電荷的比值報告質譜結果，隨後緊跟每種離子的相對豐度(在括弧中)。在Shimadzu LC / MSD電噴霧質譜儀上進行電噴霧電離（ESI）質譜分析。The reagents and solvents used below can be obtained from commercial sources. The 1 H-NMR spectrum was recorded on a Bruker 400MHZ NMR spectrometer. List of important peaks in order: multiplicity (s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; brs, wide singlet), coupling expressed in hertz (Hz) Constant (s) and number of protons. The mass spectrum results are reported as the ratio of quality to charge, followed by the relative abundance of each ion (in parentheses). Electrospray ionization (ESI) mass spectrometry analysis was performed on Shimadzu LC/MSD electrospray mass spectrometer.
合成反應Synthesis reaction
術語“溶劑”、“惰性有機溶劑”或“惰性溶劑”是指在所描述的與其有關的反應條件下呈惰性的溶劑（包括例如苯、甲苯、乙腈、四氫呋喃(“THF”)、二甲基甲醯胺（“DMF”）、乙酸乙酯（EA或EtOAc）、二氯甲烷（DCM）、二乙醚、甲醇、吡啶等）。除非另有說明，本發明反應中使用的溶劑是惰性有機溶劑，反應在惰性氣體，優選氮氣和氬氣下進行。The term "solvent", "inert organic solvent" or "inert solvent" refers to a solvent that is inert under the described reaction conditions related thereto (including, for example, benzene, toluene, acetonitrile, tetrahydrofuran ("THF"), dimethyl Formamide ("DMF"), ethyl acetate (EA or EtOAc), dichloromethane (DCM), diethyl ether, methanol, pyridine, etc.). Unless otherwise specified, the solvent used in the reaction of the present invention is an inert organic solvent, and the reaction is carried out under an inert gas, preferably nitrogen and argon.
中間體 A 的合成： 
步驟1. 在0℃下，向丙炔酸甲酯（25.0g，255mmol）的二氯甲烷（100mL）溶液中滴加嗎啉（22.2g，255mmol）。將混合物在室溫下再攪拌3小時。然後濃縮混合物，將殘渣溶於CH3 CN（300mL）中。該溶液稱為A。在0℃下，在45分鐘內小心地向4-溴-2-氯苯胺（52.8g，255mmol）的4N HCl（255mL）溶液中加入亞硝酸鈉（17.6g，255mmol）的水溶液。加完後，將溶液在0℃下再攪拌1小時。該溶液稱為B。在0℃攪拌下，將上述溶液B小心地加入到溶液C中。加完後，將混合物在室溫下再攪拌3小時。然後過濾混合物，濾餅用水洗滌並乾燥得到為黃色固體的標題產物（17.7g，21.7％）。1 H NMR (400 MHz, DMSO-d6 ): δ 14.52 (s, 0.6H), 13.23 (s, 0.4H), 9.88 (s, 0.6H), 9.55 (s, 0.4H), 7.89 (s, 1H), 7.72 - 7.65 (m, 2H), 3.86 (d, J = 15.6 Hz, 3H)。Step 1. At 0°C, add morpholine (22.2g, 255mmol) to a solution of methyl propiolate (25.0g, 255mmol) in dichloromethane (100mL) dropwise. The mixture was stirred for another 3 hours at room temperature. The mixture was then concentrated, and the residue was dissolved in CH 3 CN (300 mL). This solution is called A. At 0°C, carefully add an aqueous solution of sodium nitrite (17.6 g, 255 mmol) to a solution of 4-bromo-2-chloroaniline (52.8 g, 255 mmol) in 4N HCl (255 mL) within 45 minutes. After the addition, the solution was stirred at 0°C for another 1 hour. This solution is called B. While stirring at 0°C, the above solution B was carefully added to solution C. After the addition was complete, the mixture was stirred for another 3 hours at room temperature. The mixture was then filtered, the filter cake was washed with water and dried to give the title product (17.7 g, 21.7%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d 6 ): δ 14.52 (s, 0.6H), 13.23 (s, 0.4H), 9.88 (s, 0.6H), 9.55 (s, 0.4H), 7.89 (s, 1H), 7.72-7.65 (m, 2H), 3.86 (d, J = 15.6 Hz, 3H).
步驟2.在Ar下將2-((4-溴-2-氯苯基)二氮烯基)-3-氧代丙酸(Z)-甲基甲酯（5.00g，15.6mmol）的濃硫酸（50.0mL）溶液在100℃下攪拌10小時。將混合物冷卻至室溫並倒入冰水（200mL）中。過濾混合物，將過濾固體溶於NaOH（2M）溶液（50.0mL）中，並用乙酸乙酯（50.0mL×3）提取。然後用HCl溶液（2M）將水相調節至pH = 3,，並用乙酸乙酯（50.0mL×3）萃取。合併的有機層用Na2 SO4 乾燥，過濾並減壓濃縮得到殘餘物（2.00g），將其重結晶得到棕色固體6-溴-8-氯噌啉-3-羧酸（1.20g，26.9％）。LC-MS (ESI) [M+ H]+ 288.9.1 H NMR (400 MHz, DMSO-d6 ): δ 13.99 (s, 1H), 8.89 (s, 1H), 8.62 (d, J = 1.6 Hz, 1H), 8.51 (d, J = 2.0 Hz, 1H)。Step 2. Concentrate 2-((4-bromo-2-chlorophenyl)diazenyl)-3-oxopropionic acid (Z)-methyl methyl ester (5.00g, 15.6mmol) under Ar The sulfuric acid (50.0 mL) solution was stirred at 100°C for 10 hours. The mixture was cooled to room temperature and poured into ice water (200 mL). The mixture was filtered, and the filtered solid was dissolved in NaOH (2M) solution (50.0 mL) and extracted with ethyl acetate (50.0 mL×3). Then the aqueous phase was adjusted to pH = 3 with HCl solution (2M), and extracted with ethyl acetate (50.0 mL×3). The combined organic layer was dried over Na 2 SO 4 , filtered and concentrated under reduced pressure to obtain a residue (2.00 g), which was recrystallized to obtain a brown solid 6-bromo-8-chloro-cinoline-3-carboxylic acid (1.20 g, 26.9 %). LC-MS (ESI) [M+ H] + 288.9. 1 H NMR (400 MHz, DMSO-d 6 ): δ 13.99 (s, 1H), 8.89 (s, 1H), 8.62 (d, J = 1.6 Hz, 1H), 8.51 (d, J = 2.0 Hz, 1H).
步驟3. 在室溫下，向6-溴-8-氯噌啉-3-羧酸（380mg，1.33mmol）的MeCN（10.0mL）溶液中加入DPPA（疊氮磷酸二苯酯）(500μL)和Et3 N（350μL）。將混合物在室溫下攪拌20分鐘，然後將混合物在70℃下攪拌50分鐘。將混合物用甲苯（10.0mL）稀釋並加熱至100℃。在100℃下加入H2 O（5.00mL），並在100℃下再攪拌3小時。將混合物在室溫下冷卻並減壓濃縮。將殘餘物用H2 O（10.0mL）稀釋，並用乙酸乙酯（10.0mL×3）萃取。將合併的有機層用Na2 SO4 乾燥，過濾並減壓濃縮得到殘餘物，將其通過矽膠柱色譜法（PE：EA = 10：1-1：1，v / v）純化，得到棕色固體6-溴 -8-氯噌啉-3-胺作為中間體A（120mg，34.9％產率）。LC-MS (ESI) [M+CH3 CN]+ 257.9。Step 3. At room temperature, add DPPA (diphenyl azide phosphate) (500 μL) to a MeCN (10.0 mL) solution of 6-bromo-8-chlorocinoline-3-carboxylic acid (380 mg, 1.33 mmol) And Et 3 N (350 μL). The mixture was stirred at room temperature for 20 minutes, and then the mixture was stirred at 70°C for 50 minutes. The mixture was diluted with toluene (10.0 mL) and heated to 100°C. Add H 2 O (5.00 mL) at 100°C, and stir for another 3 hours at 100°C. The mixture was cooled at room temperature and concentrated under reduced pressure. The residue was diluted with H 2 O (10.0 mL), and extracted with ethyl acetate (10.0 mL×3). The combined organic layer was dried with Na 2 SO 4 , filtered and concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (PE:EA = 10:1 to 1:1, v/v) to obtain a brown solid 6-Bromo-8-chlorocinoline-3-amine was used as intermediate A (120 mg, 34.9% yield). LC-MS (ESI) [M+CH 3 CN] + 257.9.
6- 溴 -8- 氯 -7- 氟噌啉 -3- 胺 （中間體 B ）的合成 
步驟1. 在0℃下，將NBS（N-溴代丁二醯亞胺）（129g，1.05eq）加入到2-氯-3-氟苯胺（100g，687mmol）的DMF（500mL）溶液中。將所得混合物在室溫下攪拌16小時，然後將其倒入水（1000mL）中，對所得到的固體進行過濾，用水洗滌並乾燥得到類白色固體（140g， 91％）。LCMS (ESI) [M+H]+ 226。Step 1. At 0°C, add NBS (N-bromosuccinimide) (129 g, 1.05 eq) to a solution of 2-chloro-3-fluoroaniline (100 g, 687 mmol) in DMF (500 mL). The resulting mixture was stirred at room temperature for 16 hours, then poured into water (1000 mL), the resulting solid was filtered, washed with water and dried to obtain an off-white solid (140 g, 91%). LCMS (ESI) [M+H] + 226.
步驟2. 在0℃下，向丙炔酸乙酯（50g，510mmol）的DCM（200mL）溶液中滴加嗎啉（44.4mL，510mmol）。將混合物在室溫下再攪拌3小時。然後濃縮混合物，將殘餘物溶於CH3 CN（600mL）中。此溶液稱為C。在0℃下，在45分鐘內小心地將亞硝酸鈉（33.3g，510mmol）的水溶液（100mL）加入到4-溴-2-氯-3-氟苯胺（123g，510mmol）的4N HCl（510mL）溶液。加完後，將溶液在0℃下再攪拌1小時。此溶液稱為D。在0℃攪拌下，將上述溶液D小心地加入到溶液C中。加完後，將混合物在室溫下再攪拌3小時。然後過濾混合物，濾餅用水洗滌，並乾燥得到標題產物（100g粗品）。將粗品用EtOAc（300ml）打漿，過濾，並乾燥得到亮黃色固體（73g，38％）。LCMS (ESI) [M+H]+ 353。Step 2. At 0°C, add morpholine (44.4 mL, 510 mmol) dropwise to a solution of ethyl propiolate (50 g, 510 mmol) in DCM (200 mL). The mixture was stirred for another 3 hours at room temperature. The mixture was then concentrated, and the residue was dissolved in CH 3 CN (600 mL). This solution is called C. At 0°C, carefully add an aqueous solution (100 mL) of sodium nitrite (33.3 g, 510 mmol) to 4-bromo-2-chloro-3-fluoroaniline (123 g, 510 mmol) in 4N HCl (510 mL) within 45 minutes ) Solution. After the addition, the solution was stirred at 0°C for another 1 hour. This solution is called D. While stirring at 0°C, the above solution D was carefully added to the solution C. After the addition was complete, the mixture was stirred for another 3 hours at room temperature. The mixture was then filtered, the filter cake was washed with water, and dried to obtain the title product (100 g crude). The crude product was slurried with EtOAc (300 ml), filtered, and dried to give a bright yellow solid (73 g, 38%). LCMS (ESI) [M+H] + 353.
步驟3.在Ar下將(Z)-2-((4-溴-2-氯-3-氟苯基)二氮烯基)-3-氧代丙酸乙酯（68g，193mmol）的濃H2 SO4 （500mL）溶液在100°C下攪拌 10小時。將混合物冷卻至室溫，並倒入冰水（2000 mL）中。對混合物過濾，並乾燥得到黃色固體（53g，90％）。LCMS (ESI) [M+H]+ 307。Step 3. Concentrate (Z)-2-((4-bromo-2-chloro-3-fluorophenyl)diazenyl)-3-oxopropionate (68g, 193mmol) under Ar. The H 2 SO 4 (500 mL) solution was stirred at 100°C for 10 hours. The mixture was cooled to room temperature and poured into ice water (2000 mL). The mixture was filtered and dried to give a yellow solid (53 g, 90%). LCMS (ESI) [M+H] + 307.
步驟4. 在室溫下，將DPPA（500μL）和Et3 N （350μL）加入到6-溴-8-氯-7-氟噌啉-3-羧酸（307mg，1.00mmol）的MeCN（10.0mL）溶液中。將混合物在室溫下攪拌20分鐘，然後將混合物在70℃下攪拌50分鐘。用甲苯（10.0mL）對混合物稀釋，並加熱至100℃。在100℃下加入H2 O（5.00mL），並在100℃下再攪拌3小時。將混合物冷卻至室溫，並減壓濃縮。將殘餘物用H2 O（20.0mL）稀釋，並用乙酸乙酯（20.0mL×3）萃取。將合併的有機層用Na2 SO4 乾燥，過濾，並減壓濃縮得到殘餘物，將其通過矽膠柱色譜法（PE：EA = 10：1-1：1，v / v）純化，得到6-溴-8-氯-7-氟噌啉-3-胺（中間體 B ）。Step 4. At room temperature, add DPPA (500μL) and Et 3 N (350μL) to 6-bromo-8-chloro-7-fluorocinoline-3-carboxylic acid (307mg, 1.00mmol) in MeCN (10.0 mL) in solution. The mixture was stirred at room temperature for 20 minutes, and then the mixture was stirred at 70°C for 50 minutes. The mixture was diluted with toluene (10.0 mL) and heated to 100°C. Add H 2 O (5.00 mL) at 100°C, and stir for another 3 hours at 100°C. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was diluted with H 2 O (20.0 mL), and extracted with ethyl acetate (20.0 mL×3). The combined organic layer was dried with Na 2 SO 4 , filtered, and concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (PE:EA = 10:1-1:1, v/v) to obtain 6 -Bromo-8-chloro-7-fluorocinoline-3-amine ( Intermediate B ).
製備最終化合物的通用步驟General procedure for preparing the final compound ：:
通用步驟 1 ： 
N-(8-N-(8- 氨基Amino -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 環丙烷甲醯胺Cyclopropane carboxamide (( 化合物Compound 1)1)
步驟a. N-(6-溴-8-氯噌啉-3-基)-N-(環丙烷羰基)環丙烷甲醯胺
在室溫下，向6-溴-8-氯噌啉-3-胺（120mg，0.464mmol）的DCM（2.0mL）溶液中加入環丙基甲醯氯（121mg，1.16mmol）和Et3 N（141mg，1.39mmol）。在Ar條件下將混合物在室溫下攪拌6小時。對混合物進行減壓濃縮。將殘餘物用H2 O（10.0mL）稀釋，並用乙酸乙酯（10.0mL×3）萃取。將合併的有機層用Na2 SO4 乾燥，過濾並減壓濃縮得到殘餘物，將其通過矽膠柱色譜法（PE：EA = 10：1-1：1，v / v）純化得到為無色油的N-(6-溴-8-氯噌啉-3-基)-N-(環丙烷羰基)環丙烷甲醯胺（100mg，54.5％）。LC-MS (ESI) [M+H]+ 395.9。 At room temperature, add cyclopropylmethyl chloride (121mg, 1.16mmol) and Et 3 N to a solution of 6-bromo-8-chlorocinoline-3-amine (120mg, 0.464mmol) in DCM (2.0mL) (141mg, 1.39mmol). The mixture was stirred at room temperature for 6 hours under Ar conditions. The mixture was concentrated under reduced pressure. The residue was diluted with H 2 O (10.0 mL), and extracted with ethyl acetate (10.0 mL×3). The combined organic layer was dried with Na 2 SO 4 , filtered and concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (PE:EA = 10:1-1:1, v/v) to obtain a colorless oil N-(6-bromo-8-chlorocinoline-3-yl)-N-(cyclopropanecarbonyl)cyclopropanecarboxamide (100mg, 54.5%). LC-MS (ESI) [M+H] + 395.9.
步驟b. N-(8-氯-6-(4-甲基吡啶-3-基)噌啉-3-基)環丙烷甲醯胺
將在二氧雜環己烷（5mL）和H2 O（1.00mL）中的N-(6-溴-8-氯噌啉-3-基)-N-(環丙烷羰基)環丙烷甲醯胺（MC18-673-066-P1）（100mg，0.253mmol）、4-甲基-3- (4,4,5,5-四甲基-1,3,2-二氧雜硼烷-2-基)吡啶（66.7mg，0.304mmol）、Pd(dppf)Cl2 （37.4mg，0.051mmol）、Na2 CO3 （54.0mg，0.509mmol）的混合物在90℃和N2 下攪拌2h。將混合物冷卻至室溫，並減壓濃縮得到殘餘物，將其用H2 O（10.0mL）稀釋，並用乙酸乙酯（10.0mL×3）萃取。將合併的有機層用Na2 SO4 乾燥並減壓濃縮。通過矽膠柱色譜（PE：EA = 10：1-1：1，v / v）純化殘餘物，得到為白色油的N-(8-氯-6-(4-甲基吡啶-3-基)噌啉-3- 基)環丙烷甲醯胺（80.0mg， 93.2％）。LC-MS (ESI) [M+H]+ 339.1。Add N-(6-bromo-8-chlorocinoline-3-yl)-N-(cyclopropanecarbonyl)cyclopropane methionine in dioxane (5mL) and H 2 O (1.00mL) Amine (MC18-673-066-P1) (100mg, 0.253mmol), 4-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2 A mixture of -yl)pyridine (66.7 mg, 0.304 mmol), Pd(dppf)Cl 2 (37.4 mg, 0.051 mmol), Na 2 CO 3 (54.0 mg, 0.509 mmol) was stirred at 90° C. and N 2 for 2 h. The mixture was cooled to room temperature and concentrated under reduced pressure to obtain a residue, which was diluted with H 2 O (10.0 mL) and extracted with ethyl acetate (10.0 mL×3). The combined organic layer was dried over Na 2 SO 4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EA = 10:1 to 1:1, v/v) to obtain N-(8-chloro-6-(4-methylpyridin-3-yl) as a white oil Cinolin-3-yl)cyclopropanecarboxamide (80.0 mg, 93.2%). LC-MS (ESI) [M+H] + 339.1.
步驟c. N-(8 - ((二苯亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-基)環丙烷甲醯胺
在N2 下將在甲苯（2.00mL）和DMF（2.00mL）中的N-(8-氯-6-(4-甲基吡啶-3-基)噌啉-3-基)環丙烷甲醯胺（MC18-673-067-P1）（80mg，0.237mmol）、二苯基甲亞胺（47.2mg,，0.261mmol）、 Pd(OAc)2 （11.0mg，0.048mmol）、Xantphos（27.7mg，0.048mmol）和Cs2 CO3 （91mg，0.474mmol）的混合物在微波下於150℃下加熱3小時。將混合物冷卻至室溫，並用H2 O（10.0mL）稀釋，並用乙酸乙酯（10.0mL×3）萃取。將合併的有機層用Na2 SO4 乾燥，過濾，並減壓濃縮得到殘餘物，將其通過矽膠柱色譜法（PE：EA = 10：1-1：1，v / v）純化，得到為白色油的N-( 8-((二苯基亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-基)環丙烷甲醯胺（50.0mg，43.63％）。LC-MS (ESI) [M+H]+ 484.1。The N-(8-chloro-6-(4-methylpyridin-3-yl) cinnolin-3-yl) cyclopropane methionine in toluene (2.00 mL) and DMF (2.00 mL) under N 2 Amine (MC18-673-067-P1) (80mg, 0.237mmol), diphenylformimine (47.2mg, 0.261mmol), Pd(OAc) 2 (11.0mg, 0.048mmol), Xantphos (27.7mg, A mixture of 0.048 mmol) and Cs 2 CO 3 (91 mg, 0.474 mmol) was heated in the microwave at 150°C for 3 hours. The mixture was cooled to room temperature, diluted with H 2 O (10.0 mL), and extracted with ethyl acetate (10.0 mL×3). The combined organic layer was dried with Na 2 SO 4 , filtered, and concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (PE:EA = 10:1-1:1, v/v) to obtain White oil N-( 8-((diphenylmethylene)amino)-6-(4-methylpyridin-3-yl)cinnoline-3-yl)cyclopropanecarboxamide (50.0mg, 43.63 %). LC-MS (ESI) [M+H] + 484.1.
步驟d. N-(8-氨基-6-(4-甲基吡啶-3-基)噌啉-3-基)環丙烷甲醯胺(化合物1)
在室溫下向N-(8-((二苯基亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-基)環丙烷甲醯胺 （MC18-673-068-P1）（50.0mg，0.104mmol）的THF（3.00mL）溶液中加入HCl/二氧雜環己烷（4.0M，3.00mL）。將混合物在室溫下攪拌30min。將混合物用H2 O（10.0mL）稀釋，並用乙酸乙酯（5.00mL×3）萃取。用NaHCO3 水溶液調節水相pH = 10，並用乙酸乙酯（5.00mL X 3）萃取。將合併的有機層用Na2 SO4 乾燥，並減壓濃縮得到殘餘物，用乙醚重結晶得到為黃色固體的N-(8-氨基-6-(4-甲基吡啶-3-基)噌啉-3-基)環丙烷甲醯胺（9.96mg，30.0％產率）。LC-MS (ESI) [M+H]+ 320.1.1 H NMR (400 MHz, MeOD-d4 ): δ 8.57 (s, 1H), 8.39 (s, 2H), 7.39 - 7.36 (m, 1H), 6.95 - 6.94 (d, J = 1.2 Hz, 1H), 6.80 - 6.79 (d, J = 1.6 Hz, 1H), 2.37 (s, 3H), 2.06 - 2.00 (m, 1H), 1.04 - 1.03 (m, 2H), 0.96 - 0.94 (m, 2H)。To N-(8-((diphenylmethylene)amino)-6-(4-methylpyridin-3-yl)cinnolin-3-yl)cyclopropanecarboxamide (MC18- 673-068-P1) (50.0mg, 0.104mmol) in THF (3.00mL) was added HCl/dioxane (4.0M, 3.00mL). The mixture was stirred at room temperature for 30 min. The mixture was diluted with H 2 O (10.0 mL) and extracted with ethyl acetate (5.00 mL×3). The pH of the aqueous phase was adjusted to 10 with NaHCO 3 aqueous solution, and extracted with ethyl acetate (5.00 mL×3). The combined organic layer was dried over Na 2 SO 4 and concentrated under reduced pressure to obtain a residue, which was recrystallized from diethyl ether to obtain N-(8-amino-6-(4-methylpyridin-3-yl)) as a yellow solid. Lin-3-yl)cyclopropanecarboxamide (9.96 mg, 30.0% yield). LC-MS (ESI) [M+H] + 320.1. 1 H NMR (400 MHz, MeOD-d 4 ): δ 8.57 (s, 1H), 8.39 (s, 2H), 7.39-7.36 (m, 1H) , 6.95-6.94 (d, J = 1.2 Hz, 1H), 6.80-6.79 (d, J = 1.6 Hz, 1H), 2.37 (s, 3H), 2.06-2.00 (m, 1H), 1.04-1.03 (m , 2H), 0.96-0.94 (m, 2H).
通用 步驟A ： General Step A :
在N2 下，將在甲苯（2 mL）和DMF（2.00 mL）中的鹵化物（0.237 mmol）、二苯基甲亞胺（47.2 mg，0.261 mmol）、Pd(OAc)2 （11.0 mg，0.048 mmol）、4,5-雙(二苯基膦)-9,9-二甲基氧雜蒽（27.7 mg，0.048 mmol）和Cs2 CO3 （91 mg，0.474 mmol）的混合物在微波中於150℃加熱3 h。將混合物冷卻至室溫，用H2 O（10mL）稀釋，並用乙酸乙酯（10mL X 3）萃取。合併的有機層經Na2 SO4 乾燥，過濾並減壓濃縮得到殘餘物，將其通過矽膠柱色譜法或prep-HPLC純化，得到所需產物。Under N 2 , the halide (0.237 mmol), diphenylformimine (47.2 mg, 0.261 mmol), Pd(OAc) 2 (11.0 mg, 0.048 mmol), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (27.7 mg, 0.048 mmol) and a mixture of Cs 2 CO 3 (91 mg, 0.474 mmol) in the microwave Heat at 150°C for 3 h. The mixture was cooled to room temperature, diluted with H 2 O (10 mL), and extracted with ethyl acetate (10 mL X 3). The combined organic layer was dried over Na 2 SO 4 , filtered and concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography or prep-HPLC to obtain the desired product.
通用步驟General procedure BB ：:
在室溫下，向胺（0.923 mmol）和鹵化物（2.31 mmol）在二氧雜環己烷（10 mL）的溶液中添加t-ButylBrettPhos（225 mg，0.464 mmol），tert-BuBrettPhos-Pd-G3（131 mg，0.138 mmol）和碳酸銫（906 mg，2.78 mmol）。在Ar下，將該混合物在微波中於110℃照射2小時。將混合物冷卻至室溫。用水（50mL）稀釋混合物，用二氯甲烷（50mL X 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓下濃縮。殘餘物通過矽膠柱色譜法或prep-HPLC純化，得到所需產物。At room temperature, to a solution of amine (0.923 mmol) and halide (2.31 mmol) in dioxane (10 mL) was added t-ButylBrettPhos (225 mg, 0.464 mmol), tert-BuBrettPhos-Pd- G3 (131 mg, 0.138 mmol) and cesium carbonate (906 mg, 2.78 mmol). Under Ar, the mixture was irradiated in a microwave at 110°C for 2 hours. The mixture was cooled to room temperature. The mixture was diluted with water (50 mL) and extracted with dichloromethane (50 mL X 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue is purified by silica gel column chromatography or prep-HPLC to obtain the desired product.
通用步驟General procedure CC ：:
在室溫下，向二苯基甲亞胺化合物（0.058mmol）的MeCN（5mL）溶液中加入TFA/水（2滴/2滴）。將混合物在室溫攪拌1h。減壓除去溶劑，殘餘物用NaHCO3 溶液（30mL）稀釋，並用EA（20mL×3）萃取。有機層經無水Na2 SO4 乾燥，過濾並濃縮。殘餘物通過矽膠柱色譜法或prep-HPLC純化，得到所需化合物。At room temperature, TFA/water (2 drops/2 drops) was added to a solution of the diphenylformimine compound (0.058 mmol) in MeCN (5 mL). The mixture was stirred at room temperature for 1 h. The solvent was removed under reduced pressure, and the residue was diluted with NaHCO 3 solution (30 mL) and extracted with EA (20 mL×3). The organic layer was dried over anhydrous Na 2 SO 4 , filtered, and concentrated. The residue is purified by silica gel column chromatography or prep-HPLC to obtain the desired compound.
通用步驟General procedure DD ：:
在N2 下，將在二氧雜環己烷（5 mL）和H2 O（1mL）中的鹵化物（0.253 mmol），硼酸或硼酸酯（0.304 mmol），Pd(dppf)Cl2 （37.4 mg，0.051 mmol）和Na2 CO3 （54.0 mg，0.509 mmol）的混合物在90°C攪拌2小時。將混合物冷卻至室溫，並減壓濃縮得到殘餘物，將其用H2 O（10mL）稀釋，並用乙酸乙酯（10mL X 3）萃取。合併的有機層經Na2 SO4 乾燥，過濾並在減壓下濃縮。殘餘物通過矽膠柱色譜法或prep-HPLC純化，得到所需產物。Under N 2, the in dioxane (5 mL) and H 2 O (1mL) halide (0.253 mmol), boronic acid or ester (0.304 mmol), Pd (dppf ) Cl 2 ( A mixture of 37.4 mg, 0.051 mmol) and Na 2 CO 3 (54.0 mg, 0.509 mmol) was stirred at 90°C for 2 hours. The mixture was cooled to room temperature and concentrated under reduced pressure to obtain a residue, which was diluted with H 2 O (10 mL) and extracted with ethyl acetate (10 mL×3). The combined organic layer was dried over Na 2 SO 4, filtered and concentrated under reduced pressure. The residue is purified by silica gel column chromatography or prep-HPLC to obtain the desired product.
通用步驟General procedure EE ：:
將TFA（兩滴）加到Boc保護的化合物（0.0236mmol）在DCM（1.50mL）的溶液中，並在室溫攪拌1h。將混合物濃縮並通過prep-HPLC純化（堿，NH3 H2 O）得到所需產物。TFA (two drops) was added to a solution of Boc-protected compound (0.0236 mmol) in DCM (1.50 mL) and stirred at room temperature for 1 h. The mixture was concentrated and purified by prep-HPLC (堿, NH 3 H 2 O) to give the desired product.
通用步驟F：General Step F:
在Ar下，將鹵化物（0.0220 mmol），硼酸或硼酸酯（0.0.0266 mmol），Xphos Pd G3（3.70 mg，0.00443 mmol），Xphos（4.20 mg，0.00886 mmol）和K2 CO3 （6.10 mg，0.0443 mmol）在二氧雜環己烷/ H2 O（2 mL / 0.5 mL）的溶液攪拌至80°C ，2 h。用水（20mL）稀釋反應混合物，並用EA（20mL×3）萃取。有機層經無水Na2 SO4 乾燥，過濾，濃縮並通過prep-TLC或prep-HPLC純化，得到所需產物。Under Ar, combine halide (0.0220 mmol), boric acid or borate (0.0.0266 mmol), Xphos Pd G3 (3.70 mg, 0.00443 mmol), Xphos (4.20 mg, 0.00886 mmol) and K 2 CO 3 (6.10 mg, 0.0443 mmol) in dioxane/H 2 O (2 mL / 0.5 mL) solution was stirred to 80°C for 2 h. The reaction mixture was diluted with water (20 mL) and extracted with EA (20 mL×3). The organic layer was dried over anhydrous Na 2 SO 4 , filtered, concentrated and purified by prep-TLC or prep-HPLC to obtain the desired product.
N-(8- 氨基 -6-(4- 甲基吡啶 -3- 基 ) 噌啉 -3- 基 ) 四氫呋喃 -2- 甲醯胺 ( 化合物 2) 
中間體Intermediate CC 的製備Preparation
在N2 下，將6-溴-8-氯噌啉-3-胺（中間體 A ，194mg，0.75mmol）、4-甲基-3-(4,4,5,5-四甲基-1,3,2-二氧雜硼烷-2-基)吡啶（198mg，0.90mmol）、Pd(dppf)Cl2 （110mg，0.15mmol）和Na2 CO3 （160mg，1.5mmol）在二氧雜環己烷 （10mL）和H2 O（2.00mL）的溶液在90℃攪拌2小時。將混合物冷卻至室溫並減壓濃縮得到殘餘物，殘餘物用H2 O(10.0mL)稀釋，並用乙酸乙酯（30.0mL×3）萃取。將合併的有機層用Na2 SO4 乾燥，並減壓濃縮。通過矽膠柱色譜（PE：EA = 10：1-1：1，v / v）純化殘餘物，得到8-氯-6-(4-甲基吡啶-3-基)噌啉-3-胺（中間體 C ）。Under N 2 , 6-bromo-8-chlorocinoline-3-amine ( Intermediate A , 194mg, 0.75mmol), 4-methyl-3-(4,4,5,5-tetramethyl- 1,3,2-Dioxaborane-2-yl)pyridine (198mg, 0.90mmol), Pd(dppf)Cl 2 (110mg, 0.15mmol) and Na 2 CO 3 (160mg, 1.5mmol) in dioxy A solution of heterocyclohexane (10 mL) and H 2 O (2.00 mL) was stirred at 90°C for 2 hours. The mixture was cooled to room temperature and concentrated under reduced pressure to obtain a residue. The residue was diluted with H 2 O (10.0 mL) and extracted with ethyl acetate (30.0 mL×3). The combined organic layer was dried over Na 2 SO 4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EA = 10:1-1:1, v / v) to obtain 8-chloro-6-(4-methylpyridin-3-yl)cinnoline-3-amine ( Intermediate C ).
步驟1.在Ar和0°C下，將 DMF（1滴）和草醯氯（196mg，1.54mmol）加入到四氫呋喃-2-羧酸（36.0mg，0.308mmol）溶液中。將反應混合物在室溫下攪拌1小時。將混合物濃縮，乾燥，然後溶解在DCM（2 mL）中，並在Ar和0°C下慢慢加入到8-氯-6-(4-甲基吡啶-3-基)-噌啉-3-胺（100mg，0.370mmol）和吡啶（87.7mg，1.11mmol）在DCM（5 mL）的溶液中。將反應物在室溫下攪拌16h。用水（50mL）稀釋反應混合物，並用EA（30mL x 3）萃取。有機層用鹽水（50mL x 3）洗滌，並用無水Na2 SO4 乾燥，過濾，濃縮，通過TLC（DCM：MeOH = 30：1）純化得到為黃色油的N-(8-氯-6(4-甲基吡啶-3-基)噌啉-3-基)四氫呋喃-2-甲醯胺（80.0mg，0.217mmol，58.8%）。LC-MS (ESI) [M+H]+ 369.0。Step 1. Under Ar and 0°C, add DMF (1 drop) and oxalic chloride (196 mg, 1.54 mmol) to a solution of tetrahydrofuran-2-carboxylic acid (36.0 mg, 0.308 mmol). The reaction mixture was stirred at room temperature for 1 hour. The mixture was concentrated, dried, then dissolved in DCM (2 mL), and slowly added to 8-chloro-6-(4-methylpyridin-3-yl)-cinnoline-3 under Ar and 0°C -A solution of amine (100 mg, 0.370 mmol) and pyridine (87.7 mg, 1.11 mmol) in DCM (5 mL). The reaction was stirred at room temperature for 16 h. The reaction mixture was diluted with water (50 mL) and extracted with EA (30 mL x 3). The organic layer was washed with brine (50mL x 3), dried over anhydrous Na 2 SO 4 , filtered, concentrated, and purified by TLC (DCM:MeOH = 30:1) to obtain N-(8-chloro-6(4) as a yellow oil -Methylpyridin-3-yl)cinnolin-3-yl)tetrahydrofuran-2-carboxamide (80.0 mg, 0.217 mmol, 58.8%). LC-MS (ESI) [M+H] + 369.0.
步驟2. 根據通用步驟A 描述的方法（130o C，3h），合成為黃色固體的N-(8-((二苯亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-基)四氫呋喃-2-甲醯胺（34.3 mg， 0.067 mmol， 41.0%）。LC-MS (ESI) [M+H]+ 514.1。Step 2. According to the method described in general step A (130 o C, 3h), N-(8-((diphenylmethylene)amino)-6-(4-methylpyridine-3-) was synthesized as a yellow solid (4) Cinolin-3-yl)tetrahydrofuran-2-carboxamide (34.3 mg, 0.067 mmol, 41.0%). LC-MS (ESI) [M+H] + 514.1.
步驟3. 根據通用步驟C 描述的方法（rt，1h），合成為黃色固體的標題產物（化合物 2 ）（7.08mg，0.020mmol，34.7%產率）。LC-MS (ESI) [M+H]+ 350.0.1 H NMR (400 MHz, DMSO-d6 ): δ 10.60 (s, 1H), 8.55 (s, 1H), 8.48 (d, J = 5.2 Hz, 1H), 8.45 (s, 1H), 7.38 (d, J = 5.2 Hz, 1H), 7.02 (d, J = 1.6 Hz, 1H), 6.77 (d, J = 1.6 Hz, 1H), 6.74 (brs, 2H), 4.64 – 4.61 (m, 1H), 4.07 – 4.02 (m, 1H), 3.91 – 3.86 (m, 1H), 2.32 (s, 3H), 2.30 – 2.25 (m, 1H), 2.07 – 2.01 (m, 1H), 1.97 – 1.90 (m, 2H)。Step 3. According to the method described in General Procedure C (rt, 1h), the title product ( Compound 2 ) (7.08 mg, 0.020 mmol, 34.7% yield) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 350.0. 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.60 (s, 1H), 8.55 (s, 1H), 8.48 (d, J = 5.2 Hz , 1H), 8.45 (s, 1H), 7.38 (d, J = 5.2 Hz, 1H), 7.02 (d, J = 1.6 Hz, 1H), 6.77 (d, J = 1.6 Hz, 1H), 6.74 (brs , 2H), 4.64 – 4.61 (m, 1H), 4.07 – 4.02 (m, 1H), 3.91 – 3.86 (m, 1H), 2.32 (s, 3H), 2.30 – 2.25 (m, 1H), 2.07 – 2.01 (m, 1H), 1.97 – 1.90 (m, 2H).
(2-(4-((8- 氨基 -6-(4- 甲基吡啶 -3- 基 ) 噌啉 -3- 基 ) 氨基 )-1H- 吡唑 -1- 基 ) 丙腈 ( 化合物 3) 的制备 
步驟1. 在25℃下向8-氯-6-(4-甲基吡啶-3-基)噌啉-3-胺 （中間體C，160mg，0.592mmol）、2-(4-溴-1H-吡唑-1-基)丙腈(132mg，0.66mmol) （WO2018183964A1）、t-BuBrettPhos Pd G3 （205mg，0.237mmol）、t-BuBrettPhos（144mg，0.296mmol）的1,4-二氧雜環己烷 (20mL) 溶液中加入碳酸銫（0.97g，2.96mmol）。將混合物脫氣，並在氮氣氣氛中於120℃下加熱6小時。將反應在真空下濃縮。殘餘物通過矽膠快速色譜法純化，用二氯甲烷/甲醇（9/1）洗脫，得到2-(4-((8-氯-6-(4-甲基吡啶-3-基)噌啉-3-基)氨基)-1H-吡唑-1-基)丙腈。Step 1. Add 8-chloro-6-(4-methylpyridin-3-yl) cinnoline-3-amine (Intermediate C, 160 mg, 0.592 mmol), 2-(4-bromo-1H) at 25°C -Pyrazol-1-yl)propionitrile (132mg, 0.66mmol) (WO2018183964A1), t-BuBrettPhos Pd G3 (205mg, 0.237mmol), t-BuBrettPhos (144mg, 0.296mmol) 1,4-dioxane Cesium carbonate (0.97g, 2.96mmol) was added to the hexane (20mL) solution. The mixture was degassed and heated at 120°C for 6 hours in a nitrogen atmosphere. The reaction was concentrated under vacuum. The residue was purified by silica gel flash chromatography, eluting with dichloromethane/methanol (9/1), to give 2-(4-((8-chloro-6-(4-methylpyridin-3-yl)cinoline) -3-yl)amino)-1H-pyrazol-1-yl)propionitrile.
步驟2. 在N2 下將在甲苯（2.00mL）和DMF（2.00 mL）中的2-(4-((8-氯-6-(4-甲基吡啶-3-基)噌啉-3-基)氨基)-1H-吡唑-1-基)丙腈（80.0mg，0.206mmol）、二苯基甲亞胺（41.0mg，0.227mmol）、Pd(OAc)2 （9.2mg，0.041mmol）、4,5-雙(二苯基膦)-9,9-二甲基氧雜蒽（24.0mg，0.041mmol）和Cs2 CO3 （134mg，0.412mmol）的混合物于微波中在150℃加熱3小時。將混合物冷卻至室溫並用H2 O（10.0mL）稀釋，並用乙酸乙酯（20.0mL×3）萃取。將合併的有機層用Na2 SO4 乾燥，過濾並減壓濃縮得到殘餘物，將其通過矽膠柱色譜法（PE：EA = 10：1-1：1，v / v）純化，得到2-(4-((8 -((二苯亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-基)氨基)-1H-吡唑-1-基)丙腈。Step 2. Combine 2-(4-((8-chloro-6-(4-methylpyridin-3-yl)) cinnoline-3 in toluene (2.00 mL) and DMF (2.00 mL) under N 2 -Yl)amino)-1H-pyrazol-1-yl)propionitrile (80.0mg, 0.206mmol), diphenylformimine (41.0mg, 0.227mmol), Pd(OAc) 2 (9.2mg, 0.041mmol) ), a mixture of 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (24.0mg, 0.041mmol) and Cs 2 CO 3 (134mg, 0.412mmol) in microwave at 150℃ Heat for 3 hours. The mixture was cooled to room temperature and diluted with H 2 O (10.0 mL), and extracted with ethyl acetate (20.0 mL×3). The combined organic layer was dried with Na 2 SO 4 , filtered and concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (PE:EA = 10:1 to 1:1, v/v) to obtain 2- (4-((8 -((Diphenylmethylene)amino)-6-(4-methylpyridin-3-yl)cinnoline-3-yl)amino)-1H-pyrazol-1-yl) Propionitrile.
步驟3. 在室溫下向2-(4-((8-((二苯亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-基)氨基)-1H-吡唑-1-基)丙腈（65mg，0.122mmol）的THF（3.00mL）溶液中加入HCl/二氧雜環己烷（4.0M，3.00mL）。將混合物在室溫下攪拌30分鐘。將混合物用H2 O （10.0mL）稀釋，並用乙酸乙酯（5.00mL×3）萃取。將水相用NaHCO3 水溶液調節至pH = 10，並用乙酸乙酯（10.00mL X 3）萃取。合併的有機層用Na2 SO4 乾燥，並減壓濃縮得到殘餘物，用乙醚重結晶得到(2-(4-((8-氨基-6-(4-甲基吡啶-3-基)噌啉-吡啶-3-基)氨基)-1H-吡唑-1-基)丙腈。Step 3. To 2-(4-((8-((diphenylmethylene)amino)-6-(4-methylpyridin-3-yl)cinnolin-3-yl)amino) at room temperature To a solution of -1H-pyrazol-1-yl)propionitrile (65mg, 0.122mmol) in THF (3.00mL) was added HCl/dioxane (4.0M, 3.00mL). The mixture was stirred at room temperature for 30 minutes. The mixture was diluted with H 2 O (10.0 mL) and extracted with ethyl acetate (5.00 mL×3). The aqueous phase was adjusted to pH=10 with aqueous NaHCO 3 solution, and extracted with ethyl acetate (10.00 mL×3). The combined organic layer was dried over Na 2 SO 4 and concentrated under reduced pressure to obtain a residue, which was recrystallized from diethyl ether to obtain (2-(4-((8-amino-6-(4-methylpyridin-3-yl)) (Pyridin-3-yl)amino)-1H-pyrazol-1-yl)propionitrile.
( 反式 )-N-(8- 氨基 -6-(4- 甲基吡啶 -3- 基 ) 噌啉 -3- 基 )-2- 氰基環丙烷 -1- 甲醯胺 ( 化合物 4) 的製備 
步驟1. 將反式-2-氰基環丙烷羧酸（173mg，1.33mmol）和SOCl2 （950mg，7.98mmol）的甲苯（8 mL）溶液加熱至80°C 2h。在減壓條件下將溶劑去除。將混合物溶於DCM（2 mL）中，並在Ar和0o C下，將其加入到8-氯-6-(4-甲基吡啶-3基)噌啉-3-胺（120mg，0.440mmol）和吡啶（0.5mL）的DCM（18mL）溶液中。混合物在室溫下攪拌2h。然後用水（30mL）稀釋，用DCM（20 mL x 3）萃取。有機層用無水Na2 SO4 乾燥，過濾並濃縮得到粗產物，並用EA洗滌，過濾得到黃色固體反式-N-(8-氯-6-(4-甲基吡啶-3-基)噌啉-3基)-2-氰基環丙烷甲醯胺（110mg，0.303mmol，68.3%）。LC-MS(ESI) [M+H]+ 363.9。Step 1. Heat a solution of trans-2-cyanocyclopropane carboxylic acid (173mg, 1.33mmol) and SOCl 2 (950mg, 7.98mmol) in toluene (8 mL) to 80°C for 2h. The solvent was removed under reduced pressure. The mixture was dissolved in DCM (2 mL) and added to 8-chloro-6-(4-methylpyridin-3-yl) cinnolin-3-amine (120 mg, 0.440) under Ar and 0 o C. mmol) and pyridine (0.5 mL) in DCM (18 mL). The mixture was stirred at room temperature for 2h. Then it was diluted with water (30 mL) and extracted with DCM (20 mL x 3). The organic layer was dried with anhydrous Na 2 SO 4 , filtered and concentrated to obtain a crude product, washed with EA, and filtered to obtain a yellow solid trans-N-(8-chloro-6-(4-methylpyridin-3-yl) cinnoline -3 yl)-2-cyanocyclopropane carboxamide (110 mg, 0.303 mmol, 68.3%). LC-MS(ESI) [M+H] + 363.9.
步驟2. 根據在通用步驟A 所描述的方法（130o C ，3 h），合成為黃色油的(1R,2R)-2-氰基-N-(8-((二苯亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-基)環丙烷-1-甲醯胺（25.0mg，0.049mmol，16.2%）。LC-MS (ESI) [M+H]+ 509.0。2. The method (130 o C, 3 h) , synthesized as a yellow oil (1R, 2R) in General Procedure A described 2-cyano -N- (8 - ((diphenylmethylene) Amino)-6-(4-methylpyridin-3-yl)cinnolin-3-yl)cyclopropane-1-carboxamide (25.0 mg, 0.049 mmol, 16.2%). LC-MS (ESI) [M+H] + 509.0.
步驟3.根據通用步驟C 所描述的方法（室溫，1h），合成為黃色固體的化合物 4 （7.00mg，0.020 mmol，41.3%）。LC-MS (ESI) [M+H]+ 345.0.1 H NMR (400 MHz, DMSO-d6 ): δ 11.86 (s, 1H), 8.51 (s, 1H), 8.47 (d, J = 4.8 Hz, 1H), 8.43 (s, 1H), 7.37 (d, J = 5.2 Hz, 1H), 6.98 (d, J = 1.6 Hz, 1H), 6.76 -6.75 (m, 3H), 2.86 – 2.83 (m, 1H), 2.31 (s, 3H), 2.22 – 2.17 (m, 1H), 1.67 – 1.64 (m, 1H), 1.48 – 1.46 (m, 1H)。Step 3. According to the method described in general step C (room temperature, 1 h), compound 4 (7.00 mg, 0.020 mmol, 41.3%) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 345.0. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.86 (s, 1H), 8.51 (s, 1H), 8.47 (d, J = 4.8 Hz , 1H), 8.43 (s, 1H), 7.37 (d, J = 5.2 Hz, 1H), 6.98 (d, J = 1.6 Hz, 1H), 6.76 -6.75 (m, 3H), 2.86 – 2.83 (m, 1H), 2.31 (s, 3H), 2.22 – 2.17 (m, 1H), 1.67 – 1.64 (m, 1H), 1.48 – 1.46 (m, 1H).
N-(8- 氨基 -6-(4- 甲基吡啶 -3- 基 ) 噌啉 -3- 基 )-2- 氰基環丁烷 -1- 甲醯胺 ( 化合物 5) 
步驟1. 在0o C下向2-氰基環丁烷-1-羧酸（111mg，0.886mmol）和DMF（1滴）的DCM（3 mL）溶液中逐滴加入草醯氯（337mg，2.66mmol）。在室溫下攪拌1小時後，混合物在減壓下進行濃縮。用DCM（1mL）稀釋殘餘物，並在0o C下逐滴加入到8-氯-6-(4-甲基吡啶-3-基)噌啉-3-胺（120mg，0.443mmol）和吡啶（105mg，1.33mmol）的DCM（2mL）溶液中。混合物在室溫下攪拌16h。用水（10mL）稀釋混合物，用DCM （10 mL * 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓條件下濃縮。殘餘物用柱色譜法純化，用DCM/MeOH（100：1~20：1）洗脫得到為黃色固體的標題產物（80mg，產率：48.0%）。LC-MS (ESI) [M+H]+ 378.0。1. Step-wise a solution of 2-cyano-cyclobutane-1-carboxylic acid (111mg, 0.886mmol) and DMF (1 drop) in DCM (3 mL) at 0 o C was added dropwise oxalyl acyl chloride (337mg, 2.66mmol). After stirring at room temperature for 1 hour, the mixture was concentrated under reduced pressure. Diluted with DCM (1mL) residue, and added dropwise at 0 o C was added to 8-chloro-6- (4-methyl-pyridin-3-yl) cinnoline-3-amine (120mg, 0.443mmol) and pyridine (105mg, 1.33mmol) in DCM (2mL). The mixture was stirred at room temperature for 16 h. The mixture was diluted with water (10 mL) and extracted with DCM (10 mL * 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography and eluted with DCM/MeOH (100:1-20:1) to obtain the title product (80mg, yield: 48.0%) as a yellow solid. LC-MS (ESI) [M+H] + 378.0.
步驟2. 根據通用步驟A 描述的方法（150o C ，3h），合成為黃色固體的N-(8-((二苯亞甲基)氨基)-6(4-甲基吡啶-3-基)噌啉-3-基)-2-氰基環丁烷-1-甲醯胺（3.00mg，21.7%）。LC-MS (ESI) [M+H]+ 523.0。Step 2. According to the method described in general step A (150 o C, 3h), N-(8-((diphenylmethylene)amino)-6(4-methylpyridin-3-yl) was synthesized as a yellow solid ) Cinolin-3-yl)-2-cyanocyclobutane-1-carboxamide (3.00 mg, 21.7%). LC-MS (ESI) [M+H] + 523.0.
步驟3. 根據通用步驟C 描述的方法，合成為黃色固體的化合物 5 （2.08mg，0.006mmol，20.2%）。LC-MS (ESI) [M+H]+ 359.0.1 H NMR (400 MHz, DMSO-d6 ): δ 11.41 (s, 1H), 8.58 (s, 1H), 8.47 (d, J = 5.2 Hz, 1H), 8.44 (s, 1H), 7.37 (d, J = 4.8 Hz, 1H), 7.00 (d, J = 1.6 Hz, 1H), 6.76 (d, J = 1.2 Hz, 1H), 6.72 (brs, 2H), 3.89 – 3.82 (m, 1H), 3.67 – 3.58 (m, 1H), 2.32 (s, 3H), 2.26 – 2.21 (m, 4H)。Step 3. According to the method described in general step C , compound 5 (2.08 mg, 0.006 mmol, 20.2%) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 359.0. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.41 (s, 1H), 8.58 (s, 1H), 8.47 (d, J = 5.2 Hz , 1H), 8.44 (s, 1H), 7.37 (d, J = 4.8 Hz, 1H), 7.00 (d, J = 1.6 Hz, 1H), 6.76 (d, J = 1.2 Hz, 1H), 6.72 (brs , 2H), 3.89 – 3.82 (m, 1H), 3.67 – 3.58 (m, 1H), 2.32 (s, 3H), 2.26 – 2.21 (m, 4H).
2-((8- 氨基 -6-(4- 甲基吡啶 -3- 基 ) 噌啉 -3- 基 ) 氨基 )-6- 異丙基 -5,6- 二氫 -4H- 吡唑並 [1,5-d] [1 ,4] 二氮雜卓 -7(8H)- 酮 ( 化合物 6) 
步驟1. 根據通用步驟B 描述的方法（110o C下2h），合成為黃色固體的2-((8-氯-6-(4-甲基吡啶-3-基)噌啉-3-基)氨基)-6-異丙基-5,6-二氫-4H-吡唑並[1,5-d] [1 ,4] 二氮雜卓-7(8H)-酮（95.0mg，22.2%）。LC-MS (ESI) [M+H]+ 462.0.1 H NMR (400 MHz, DMSO-d6 ): δ 10.33 (s, 1H), 8.53 - 8.50 (m, 2H), 8.34 (s, 1H), 7.93 (d, J = 1.2 Hz, 1H), 7.79 (d, J = 1.6 Hz, 1H), 7.41 (d, J = 5.2 Hz, 1H), 6.06 (s, 1H), 5.03 (s, 2H), 4.61 - 4.57 (m, 1H), 3.82 - 3.78 (m, 2H), 3.03 - 3.00 (m, 2H), 2.36 (s, 3H), 1.12 (d, J = 6.8 Hz, 6H)。Step 1. According to the method described in general step B (2h at 110 o C), 2-((8-chloro-6-(4-methylpyridin-3-yl)cinolin-3-yl) was synthesized as a yellow solid )Amino)-6-isopropyl-5,6-dihydro-4H-pyrazolo[1,5-d] [1 ,4]diazepine-7(8H)-one (95.0mg, 22.2 %). LC-MS (ESI) [M+H] + 462.0. 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.33 (s, 1H), 8.53-8.50 (m, 2H), 8.34 (s, 1H) , 7.93 (d, J = 1.2 Hz, 1H), 7.79 (d, J = 1.6 Hz, 1H), 7.41 (d, J = 5.2 Hz, 1H), 6.06 (s, 1H), 5.03 (s, 2H) , 4.61-4.57 (m, 1H), 3.82-3.78 (m, 2H), 3.03-3.00 (m, 2H), 2.36 (s, 3H), 1.12 (d, J = 6.8 Hz, 6H).
步驟2. 根據通用步驟A 描述的方法（在微波下，130o C ，3h），合成為黃色固體的2-((8-(( 二苯亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-基)氨基)-6-異丙基-5,6-二氫-4H-吡唑並[1,5-d] [1 ,4] 二氮雜卓-7(8H)-酮（14.0mg，26.6%）。LC-MS (ESI) [M+H]+ 607.2。Step 2. According to the method described in general step A (under microwave, 130 o C, 3h), 2-((8-((diphenylmethylene)amino)-6-(4-methyl) was synthesized as a yellow solid (Pyridin-3-yl) cinnolin-3-yl) amino)-6-isopropyl-5,6-dihydro-4H-pyrazolo[1,5-d] [1 ,4]diazepine Zhuo-7(8H)-one (14.0mg, 26.6%). LC-MS (ESI) [M+H] + 607.2.
步驟3. 根據通用步驟C 描述的方法（室溫，30min），合成為黃色固體的化合物 6 （9.06 mg, 59.2%）。LC-MS (ESI) [M+H]+ 443.2.1 H NMR (400 MHz, DMSO-d6 ): δ 9.85 (s, 1H), 8.44 (s, 2H), 8.03 (s, 1H), 7.36 (s, 1H), 6.83 (s, 1H), 6.53 (s, 3H), 6.04 (s, 1H), 5.00 (s, 2H), 4.59 (s, 1H), 3.79 (s, 2H), 3.00 (s, 2H), 2.30 (s, 3H), 1.12 (d, J = 4.4 Hz, 6H)。Step 3. According to the method described in general step C (room temperature, 30 min), compound 6 (9.06 mg, 59.2%) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 443.2. 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.85 (s, 1H), 8.44 (s, 2H), 8.03 (s, 1H), 7.36 (s, 1H), 6.83 (s, 1H), 6.53 (s, 3H), 6.04 (s, 1H), 5.00 (s, 2H), 4.59 (s, 1H), 3.79 (s, 2H), 3.00 ( s, 2H), 2.30 (s, 3H), 1.12 (d, J = 4.4 Hz, 6H).
N-(8-N-(8- 氨基Amino -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )-1-()-1-( 氰甲基Cyanomethyl )) 氮雜環丁烷Azetidine -2--2- 甲醯胺Formamide (( 化合物Compound 7)7)
根據通用步驟2，通過中間體C和1-(氰甲基)氮雜環丁烷-2-碳醯氯製備N-(8-氨基-6-(4-甲基吡啶-3-基)噌啉-3-基)-1-(氰甲基)氮雜環丁烷-2-甲醯胺，其中1-(氰甲基)氮雜環丁烷-2-碳醯氯以與化合物 2 相同的方式由1-(氰甲基)氮雜環丁烷-2-羧酸和亞硫醯氯在原位生成。According to general procedure 2, prepare N-(8-amino-6-(4-methylpyridin-3-yl)) via intermediate C and 1-(cyanomethyl)azetidine-2-carbochloride Lin-3-yl)-1-(cyanomethyl)azetidine-2-carboxamide, of which 1-(cyanomethyl)azetidine-2-carboxamide is the same as compound 2 The method is generated in situ from 1-(cyanomethyl)azetidine-2-carboxylic acid and sulfite chloride.
N-(8-N-(8- 氨基Amino -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )-1-(1-)-1-(1- 甲基methyl -1H--1H- 吡唑Pyrazole -4--4- 基base )) 氮雜環丁烷Azetidine -2--2- 甲醯胺Formamide (( 化合物Compound 8)8)
根據通用步驟2，經由中間體C和由1-(1-甲基-1H-吡唑-4-基)氮雜環丁烷-2-碳醯氯製備N-(8-氨基-6-(4-甲基吡啶-3-基)噌啉-3-基)-1-(1-甲基-1H-吡唑-4-基)氮雜環丁烷-2-甲醯胺，1-(1-甲基-1H-吡唑-4-基)氮雜環丁烷-2-碳醯氯是利用1-(1-甲基-1H-吡唑-4-基)氮雜環丁烷-2-羧酸（由PCT Int.Appl. 2010147791報導的通過CuI催化的氮雜環丁烷-2-羧酸與4-溴-1-甲基-1H-吡唑的偶聯反應製備）和亞硫醯氯在原位反應得到。According to general procedure 2, N-(8-amino-6-( 4-methylpyridin-3-yl) cinnolin-3-yl)-1-(1-methyl-1H-pyrazol-4-yl)azetidine-2-carboxamide, 1-( 1-Methyl-1H-pyrazol-4-yl)azetidine-2-carbochloride is the use of 1-(1-methyl-1H-pyrazol-4-yl)azetidine- 2-carboxylic acid (prepared by CuI-catalyzed coupling reaction of azetidine-2-carboxylic acid and 4-bromo-1-methyl-1H-pyrazole reported by PCT Int.Appl. 2010147791) and Thiochlorine is obtained by reaction in situ.
N-(8-N-(8- 氨基Amino -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 四氫Tetrahydro -2H--2H- 吡喃Pyran -2--2- 甲醯胺Formamide (( 化合物Compound 9)9)
按照通用步驟2，經由中間體 C 和四氫-2H-吡喃-2-碳醯氯（Packiarajan, Mathivanan et al, Bioorganic & Medicinal Chemistry Letters, 23(14), 4037-4043; 2013）反應製備N-(8-氨基-6-(4-甲基吡啶-3-基)噌啉-3-基)四氫-2H-吡喃-2-甲醯胺。According to general procedure 2, N was prepared by reaction of intermediate C and tetrahydro-2H-pyran-2-carbochloride (Packiarajan, Mathivanan et al, Bioorganic & Medicinal Chemistry Letters, 23(14), 4037-4043; 2013) -(8-Amino-6-(4-methylpyridin-3-yl)cinolin-3-yl)tetrahydro-2H-pyran-2-carboxamide.
N-(8- 氨基 -6-(4- 甲基吡啶 -3- 基 ) 噌啉 -3- 基 ) 四氫 -2H- 吡喃 -3- 甲醯胺 ( 化合物 10) 
步驟1. 將四氫-2H-呋喃-3-羧酸的SOCl2 （2mL）溶液加熱到80°C 2h。冷卻至室溫後，在室溫減壓條件下去除溶劑。將混合物溶解在DCM（2mL）中，並在0o C和Ar下加入到8-氯-6-(4-甲基吡啶-3-基)噌啉-3-氨（100mg，0.370mmol）和TEA（0.8mL）的DCM（5mL）溶液中。混合物在室溫下攪拌16h。然後用水（30mL）稀釋，用DCM (20 mL x 3)萃取。有機層用無水Na2 SO4 乾燥，過濾，濃縮，並用矽膠柱色譜法（DCM:MeOH=50:1）純化得到黃色固體N-(8-氯-6-(4-甲基吡啶-3-基)噌啉-3-基)四氫-2H-呋喃-3-甲醯胺（72.7mg，0.191mmol，51.5%）。LC-MS (ESI) [M+H]+ 383.0。 Step 1. Heat the SOCl 2 (2mL) solution of tetrahydro-2H-furan-3-carboxylic acid to 80°C for 2h. After cooling to room temperature, the solvent was removed under reduced pressure at room temperature. The mixture was dissolved in DCM (2mL), was added to 8-chloro and at 0 o C and Ar -6- (4- methyl-pyridin-3-yl) cinnoline-3-amino (100mg, 0.370mmol) and TEA (0.8mL) in DCM (5mL) solution. The mixture was stirred at room temperature for 16 h. Then it was diluted with water (30 mL) and extracted with DCM (20 mL x 3). The organic layer was dried over anhydrous Na 2 SO 4 , filtered, concentrated, and purified by silica gel column chromatography (DCM:MeOH=50:1) to obtain a yellow solid N-(8-chloro-6-(4-methylpyridine-3- (4) Cinolin-3-yl) tetrahydro-2H-furan-3-carboxamide (72.7 mg, 0.191 mmol, 51.5%). LC-MS (ESI) [M+H] + 383.0.
步驟2. 根據通用步驟A描述的方法（150o C，3h），合成為黃色油狀物的N-(8-((二苯亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-基)四氫-2H-呋喃-3-甲醯胺（14.9mg，0.028mmol，15.4%）。LC-MS (ESI) [M+H]+ 528.1。Step 2. According to the method described in general step A (150 o C, 3h), N-(8-((diphenylmethylene)amino)-6-(4-methylpyridine-) was synthesized as a yellow oil 3-yl) Cinolin-3-yl) tetrahydro-2H-furan-3-carboxamide (14.9 mg, 0.028 mmol, 15.4%). LC-MS (ESI) [M+H] + 528.1.
步驟3.根據通用步驟C描述的方法（室溫，1h），合成為黃色固體的化合物 10 （1.18 mg， 0.003 mmol，14.3 %）。LC-MS (ESI) [M+H]+ 364.1.1 H NMR (400 MHz, MeOD-d4 ): δ 8.60 (s, 1H), 8.41 (br s, 1H), 7.40 (d, J = 4.8 Hz, 1H), 6.97 (d, J = 1.2 Hz, 1H), 6.81 (d, J = 1.6 Hz, 1H), 4.12 - 4.09 (m, 1H), 3.92 - 3.89 (m, 1H), 3.69 - 3.64 (m, 1H), 3.54 - 3.47 (m, 1H), 2.93 - 2.89 (m, 1H), 2.38 (s, 3H), 2.18 - 2.08 (m, 1H),1.94 - 1.89 (m, 1H), 1.79 - 1.73 (m, 2H)。Step 3. According to the method described in general step C (room temperature, 1 h), compound 10 (1.18 mg, 0.003 mmol, 14.3%) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 364.1. 1 H NMR (400 MHz, MeOD-d 4 ): δ 8.60 (s, 1H), 8.41 (br s, 1H), 7.40 (d, J = 4.8 Hz, 1H), 6.97 (d, J = 1.2 Hz, 1H), 6.81 (d, J = 1.6 Hz, 1H), 4.12-4.09 (m, 1H), 3.92-3.89 (m, 1H), 3.69-3.64 (m, 1H), 3.54-3.47 (m, 1H), 2.93-2.89 (m, 1H), 2.38 (s, 3H), 2.18-2.08 (m, 1H), 1.94-1.89 (m, 1H), 1.79 -1.73 (m, 2H).
N-(8-N-(8- 氨基Amino -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 四氫呋喃Tetrahydrofuran -3--3- 甲醯胺Formamide (( 化合物Compound 11)11)
利用與製備化合物 10 的相似的方法製備黃色固體化合物 11 （8.71mg，32.2%產率）。LC-MS (ESI) [M+H]+ 350.2.1 H NMR (400 MHz, DMSO-d6 ): δ 11.38 (s, 1H), 8.57 (s, 1H), 8.48 - 8.44 (m, 2H), 7.37 (d, J = 4.8 Hz, 1H), 6.99 (d, J = 1.2 Hz, 1H), 6.75 - 6.71 (m, 3H), 4.02 - 3.97 (m, 1H), 3.82 - 3.73 (m, 3H), 3.47 - 3.44 (m, 1H), 2.31 (s, 3H), 2.17 - 2.12 (m, 3H)。 A yellow solid compound 11 (8.71 mg, 32.2% yield) was prepared using a method similar to that for preparing compound 10. LC-MS (ESI) [M+H] + 350.2. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.38 (s, 1H), 8.57 (s, 1H), 8.48-8.44 (m, 2H) , 7.37 (d, J = 4.8 Hz, 1H), 6.99 (d, J = 1.2 Hz, 1H), 6.75-6.71 (m, 3H), 4.02-3.97 (m, 1H), 3.82-3.73 (m, 3H) ), 3.47-3.44 (m, 1H), 2.31 (s, 3H), 2.17-2.12 (m, 3H).
( 反式 )-N-(8- 氨基 -6-(8- 甲基 -2,3- 二氫 -1H- 吡啶並 [2,3- b] [1,4] 惡嗪 -7- 基 ) 噌啉 -3- 基 )-2-(1- 甲基 -1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺 ( 化合物 12) 的製備 
步驟1.根據化合物 4 中相似的步驟所描述的方法，合成為黃色固體的7-(8-氯-3-((1R,2R)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-羰基)噌啉-6-基)-8-甲基-2,3-二氫-1H-吡啶並[2,3-b][1,4]惡嗪-1-羧酸叔丁基酯和7-(8-氯-3-((1S,2S)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-羰基)噌啉-6-基)-8-甲基-2,3-二氫-1H-吡啶並[2,3-b][1,4]惡嗪-1-羧酸叔丁基酯的外消旋混合物（130mg，56.8%）。LC-MS (ESI) [M+H]+ 574.2。Step 1. According to the method described in the similar step in compound 4 , 7-(8-chloro-3-((1R,2R)-2-(1-methyl-1H-pyrazole-4) was synthesized as a yellow solid -Yl)cyclopropane-1-carbonyl)cinnoline-6-yl)-8-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1 -Tert-butyl carboxylate and 7-(8-chloro-3-((1S,2S)-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-carbonyl)cinoline -6-yl)-8-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylate in racemic mixture (130mg, 56.8%). LC-MS (ESI) [M+H] + 574.2.
步驟2. 根據通用步驟A 描述的方法（130o C，3h），合成為黃色固體的7-(8-((二苯亞甲基)氨基)-3-(-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-羰基)噌啉-6-基)-8-甲基-2,3-二氫-1H-吡啶並[2,3-b][1,4]惡嗪-1-羧酸叔丁基酯（30mg，24.0%）。LC-MS (ESI) [M+H]+ 721.1。Step 2. According to the method described in general step A (130 o C, 3h), 7-(8-((diphenylmethylene)amino)-3-(-2-(1-methyl) was synthesized as a yellow solid -1H-pyrazol-4-yl)cyclopropane-1-carbonyl)cinnoline-6-yl)-8-methyl-2,3-dihydro-1H-pyrido[2,3-b][1 , 4] tert-butyl oxazine-1-carboxylate (30 mg, 24.0%). LC-MS (ESI) [M+H] + 721.1.
步驟3. 根據通用步驟C 描述的方法，合成為黃色固體的7-(8-氨基-3-(-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-羰基)噌啉-6-基)-8-甲基-2,3-二氫-1H-吡啶並[2,3-b][1,4]惡嗪-1-羧酸叔丁基酯（20mg，87.0%）。LC-MS (ESI) [M+H]+ 557.3。Step 3. According to the method described in general step C , 7-(8-amino-3-(-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-carbonyl) was synthesized as a yellow solid ) Cinolin-6-yl)-8-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl ester (20mg , 87.0%). LC-MS (ESI) [M+H] + 557.3.
步驟4. 根據通用步驟E 描述的方法，合成為黃色固體的標題產物（69）（8.52mg，57.8%）。LC-MS (ESI) [M+H]+ 457.2.1 H NMR (400 MHz, DMSO-d6 ): δ 11.48 (s, 1H), 8.51 (s, 1H), 7.58 (s, 1H), 7.35 (s, 1H), 7.31 (s, 1H), 6.84 (s, 1H), 6.68 (s, 1H), 6.62 (s, 2H), 5.67 (s, 1H), 4.27 (s, 2H), 3.78 (s, 3H), 3.36 - 3.32 (m, 2H), 2.30 - 2.23 (m, 2H), 2.03 (s, 3H), 1.43 - 1.41 (m, 1H), 1.23 (s, 1H)。Step 4. According to the method described in General Procedure E , the title product (69) (8.52 mg, 57.8%) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 457.2. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.48 (s, 1H), 8.51 (s, 1H), 7.58 (s, 1H), 7.35 (s, 1H), 7.31 (s, 1H), 6.84 (s, 1H), 6.68 (s, 1H), 6.62 (s, 2H), 5.67 (s, 1H), 4.27 (s, 2H), 3.78 ( s, 3H), 3.36-3.32 (m, 2H), 2.30-2.23 (m, 2H), 2.03 (s, 3H), 1.43-1.41 (m, 1H), 1.23 (s, 1H).
2-((8- 氨基 -6-(8- 甲基 -2,3- 二氫 -1H- 吡啶並 [2,3-b] [1,4] 惡嗪 -7- 基 ) 噌啉 -3- 基 ) 氨基 )-6- 異丙基 -5,6- 二氫 -4H- 吡唑並 [1,5-d] [1,4] 二氮雜卓 -7(8H)- 酮 ( 化合物 13) 
步驟1. 根據通用步驟A ，合成為黃色固體的中間體 D （80mg，0.187mmol，48.3%產率）。LC-MS (ESI) [M+H]+ 428.0。Step 1. According to general procedure A , intermediate D (80 mg, 0.187 mmol, 48.3% yield) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 428.0.
步驟2. 根據通用步驟 B 描述的方法（經由中間體 D 和根據WO2018183964A1製備的已知的化合物2-溴-6-異丙基-5,6-二氫-4H-吡唑並[1,5-d] [1,4]二氮雜卓-7(8H)-酮，），合成為黃色固體的7-(8-氯-3-((6-異丙基-7-氧代-5,6,7,8-四氫-4H-吡唑並[1,5-d][1,4]二苯氮卓-2-基)氨基)噌啉-6-基)-8-甲基-2,3-二氫-1H-吡啶並[2,3-b][1,4]惡嗪-1-羧酸叔丁基酯（58mg，0.0939mmol，26.7%）。LC-MS (ESI) [M+H]+ 619.2。Step 2. According to the method described in General Procedure B (via Intermediate D and the known compound 2-bromo-6-isopropyl-5,6-dihydro-4H-pyrazolo[1,5 -d] [1,4]diazepine-7(8H)-one,), synthesized as a yellow solid 7-(8-chloro-3-((6-isopropyl-7-oxo-5 ,6,7,8-Tetrahydro-4H-pyrazolo[1,5-d][1,4]diazepine-2-yl)amino)cinoline-6-yl)-8-methyl -2,3-Dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl ester (58 mg, 0.0939 mmol, 26.7%). LC-MS (ESI) [M+H] + 619.2.
步驟3. 根據通用步驟 A 描述的方法，合成為黃色固體的2-((8-二苯亞甲基)氨基)-6-(8-甲基-2,3-二氫-1H-吡啶並[2,3-b][1,4]惡嗪-7-基)噌啉-3-基)氨基)-6-異丙基-5,6-二氫-4H-吡唑並[1,5-d][1,4]二氮雜卓-7(8H)-酮（17.0mg，0.0256mmol，28.8%）。LC-MS (ESI) [M+H]+ 664.1。Step 3. According to the method described in general step A , 2-((8-diphenylmethylene)amino)-6-(8-methyl-2,3-dihydro-1H-pyrido) was synthesized as a yellow solid. [2,3-b][1,4]oxazine-7-yl)cinolin-3-yl)amino)-6-isopropyl-5,6-dihydro-4H-pyrazolo[1, 5-d][1,4]diazepine-7(8H)-one (17.0mg, 0.0256mmol, 28.8%). LC-MS (ESI) [M+H] + 664.1.
步驟4. 根據通用步驟 C 描述的方法，合成為黃色固體的化合物 13 （1.43 mg, 0.00287 mmol, 11.2 %）。LC-MS (ESI) [M+H]+ 500.2.1 H NMR (400 MHz, MeOD-d4 ): δ 8.00 (s, 1H), 7.38 (s, 1H), 6.79 (d, J = 1.6 Hz, 1H), 6.59 (d, J = 1.6 Hz, 1H), 6.06 (s, 1H), 5.06 (s, 1H), 4.77 - 4.74 (m, 1H), 4.57 (s, 1H), 4.39 - 4.36 (m, 2H), 3.88 - 3.85 (m, 2H), 2.49 - 3.47 (m, 2H), 3.13 - 3.11 (m, 2H), 2.11 (s, 3H), 1.22 (d, J = 6.8 Hz, 6H)。Step 4. According to the method described in general step C , compound 13 (1.43 mg, 0.00287 mmol, 11.2 %) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 500.2. 1 H NMR (400 MHz, MeOD-d 4 ): δ 8.00 (s, 1H), 7.38 (s, 1H), 6.79 (d, J = 1.6 Hz , 1H), 6.59 (d, J = 1.6 Hz, 1H), 6.06 (s, 1H), 5.06 (s, 1H), 4.77-4.74 (m, 1H), 4.57 (s, 1H), 4.39-4.36 ( m, 2H), 3.88-3.85 (m, 2H), 2.49-3.47 (m, 2H), 3.13-3.11 (m, 2H), 2.11 (s, 3H), 1.22 (d, J = 6.8 Hz, 6H) .
2-(4-((8-2-(4-((8- 氨基Amino -6-(8--6-(8- 甲基methyl -2,3--2,3- 二氫Dihydro -1H--1H- 吡啶並Pyrido [2,3-b] [1,4][2,3-b] [1,4] 惡嗪Oxazine -7--7- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-1H-)-1H- 吡唑Pyrazole -1--1- 基base )) 丙腈Propionitrile (( 化合物Compound 14)14)
根據化合物 13 相似的方法，經由中間體E和2-(4-溴-1H-吡唑-1-基)丙腈（WO2018183964A1）製備2-(4-((8-氨基-6-(8-甲基-2,3-二氫-1H-吡啶並[2,3-b] [1,4]惡嗪-7-基)噌啉-3-基)氨基)-1H-吡唑-1-基)丙腈。According to a similar method to compound 13 , through intermediate E and 2-(4-bromo-1H-pyrazol-1-yl)propionitrile (WO2018183964A1), 2-(4-((8-amino-6-(8- Methyl-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazin-7-yl)cinolin-3-yl)amino)-1H-pyrazole-1-基)Propionitrile.
2-((8-2-((8- 氨基Amino -6-(5--6-(5- 氨基Amino -4--4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-6-)-6- 異丙基Isopropyl -5,6--5,6- 二氫Dihydro -4H--4H- 吡唑並Pyrazolo [1,5-d] [1,4][1,5-d] [1,4] 二氮雜卓Diazepam -7(8H)--7(8H)- 酮ketone (( 化合物Compound 15)15)
根據化合物 13 相似的方法，經由(5-(雙(叔丁氧基羰基)氨基)-4-甲基吡啶-3-基)硼酸（WO2018183964A1）和2-溴-6-異丙基-5,6-二氫-4H-吡唑並[1,5-d] [1,4]二氮雜卓-7(8H)-酮（WO2018183964A1）製備2-((8-氨基-6-(5-氨基-4-甲基吡啶-3-基)噌啉-3-基)氨基)-6-異丙基-5,6-二氫-4H-吡唑並[1,5-d] [1,4]二氮雜卓-7(8H)-酮。According to a similar method to compound 13 , via (5-(bis(tert-butoxycarbonyl)amino)-4-methylpyridin-3-yl)boronic acid (WO2018183964A1) and 2-bromo-6-isopropyl-5, 6-Dihydro-4H-pyrazolo[1,5-d] [1,4]diazepine-7(8H)-one (WO2018183964A1) Preparation of 2-((8-amino-6-(5- Amino-4-methylpyridin-3-yl)cinnoline-3-yl)amino)-6-isopropyl-5,6-dihydro-4H-pyrazolo[1,5-d] [1, 4] Diazepam-7(8H)-one.
(( 反式Trans )-N-(8-)-N-(8- 氨基Amino -6-(5--6-(5- 甲氧基Methoxy -4--4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )-2-)-2- 氰基環丙烷Cyanocyclopropane -1--1- 甲醯胺Formamide (( 化合物Compound 16)16)
根據化合物 13 相似的方法，經由(5-甲氧基-4-甲基吡啶-3-基)硼酸（Biagetti, Matteo et al, US20150166549 A1）製備化合物 16 。According to a similar method to compound 13 , compound 16 was prepared via (5-methoxy-4-methylpyridin-3-yl)boronic acid (Biagetti, Matteo et al, US20150166549 A1).
2-((8-2-((8- 氨基Amino -6-(5--6-(5- 甲氧基Methoxy -4--4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-6-)-6- 異丙基Isopropyl -5,6--5,6- 二氫Dihydro -4H--4H- 吡唑並Pyrazolo [1,5- d] [1,4][1,5- d] [1,4] 二氮雜卓Diazepam -7(8H)--7(8H)- 酮ketone (( 化合物Compound 17)17)
根據化合物 13 相似的方法，通過中間體F和2-溴-6-異丙基-5,6-二氫-4H-吡唑並[1,5-d] [1,4]二氮雜卓-7(8H)-酮（WO2018183964A1）製備2-((8-氨基-6-(5-甲氧基-4-甲基吡啶-3-基)噌啉-3-基)氨基)-6-異丙基-5,6-二氫-4H-吡唑並[1,5- d] [1,4]二氮雜卓-7(8H)-酮。According to a similar method to compound 13 , through intermediate F and 2-bromo-6-isopropyl-5,6-dihydro-4H-pyrazolo[1,5-d] [1,4]diazepine -7(8H)-ketone (WO2018183964A1) Preparation of 2-((8-amino-6-(5-methoxy-4-methylpyridin-3-yl)cinolin-3-yl)amino)-6- Isopropyl-5,6-dihydro-4H-pyrazolo[1,5- d] [1,4]diazepine-7(8H)-one.
6-(8-6-(8- 氨基Amino -3-((6--3-((6- 異丙基Isopropyl -7--7- 氧代Oxo -5,6,7,8--5,6,7,8- 四氫Tetrahydro -4H--4H- 吡唑並Pyrazolo [1,5-d] [1,4][1,5-d] [1,4] 二氮雜卓Diazepam -2--2- 基base )) 氨基Amino )) 噌啉Cinolin -6--6- 基base )-5-)-5- 甲基苯並Methyl benzo [d][d] 惡唑Oxazole -2(3H)--2(3H)- 酮ketone (( 化合物Compound 18)18)
根據化合物17相似的方法，經由(5-甲基-2-氧代-2,3-二氫苯丙[d]惡唑-6-基)硼酸（WO2018183964A1）製備化合物 18 。 According to a similar method to compound 17, compound 18 was prepared via (5-methyl-2-oxo-2,3-dihydrophenylprop[d]oxazol-6-yl)boronic acid (WO2018183964A1).
(( 反式Trans )-N-(8-)-N-(8- 氨基Amino -6-(5--6-(5- 甲基methyl -2--2- 氧代Oxo -2,3--2,3- 二氫苯並Dihydrobenzo [d][d] 惡唑Oxazole -6--6- 基base )) 噌啉Cinolin -3--3- 基base )-2-)-2- 氰基環丙烷Cyanocyclopropane -1--1- 甲醯胺Formamide (( 化合物Compound 19)19)
根據通用步驟1，通過中間體A、(5-甲基-2-氧代-2,3-二氫苯並[d]惡唑-6-基)硼酸（WO2018183964A1）和2-氰基環丙烷-1-碳醯氯製備化合物 19 。According to general procedure 1, through intermediate A, (5-methyl-2-oxo-2,3-dihydrobenzo[d]oxazol-6-yl)boronic acid (WO2018183964A1) and 2-cyanocyclopropane -1-Carbochloride to prepare compound 19 .
(( 反式Trans )-N-(8-)-N-(8- 氨基Amino -6-(5-(-6-(5-( 二甲基磷醯基Dimethylphosphoryl )-2-)-2- 甲基苯基Methyl phenyl )) 噌啉Cinolin -3--3- 基base )-2-)-2- 氰基環丙烷Cyanocyclopropane -1--1- 甲醯胺Formamide (( 化合物Compound 20)20)
N-(8-N-(8- 氨基Amino -6-(5-(-6-(5-( 二甲基磷醯基Dimethylphosphoryl )-2-)-2- 甲基苯基Methyl phenyl )) 噌啉Cinolin -3--3- 基base )-1-(1-)-1-(1- 甲基methyl -1H--1H- 吡唑Pyrazole -4--4- 基base )) 氮雜環丁烷Azetidine -2--2- 甲醯胺Formamide (( 化合物Compound 21)twenty one)
2-((8-2-((8- 氨基Amino -6-(5-(-6-(5-( 二甲基磷醯基Dimethylphosphoryl )-2-)-2- 甲基苯基Methyl phenyl )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-6-)-6- 異丙基Isopropyl -5,6--5,6- 二氫Dihydro -4H--4H- 吡唑並Pyrazolo [1,5-d] [1,4][1,5-d] [1,4] 二氮雜Diaza -7(8H)--7(8H)- 酮ketone (( 化合物Compound 22)twenty two)
根據通用步驟A-F製備化合物 20-22 。 Compound 20-22 was prepared according to general procedure AF .
N-(8- 氨基 -7- 氟 -6-(4- 甲基吡啶 -3- 基 ) 噌啉 -3- 基 ) 環丙烷甲醯胺 ( 化合物 23) 
步驟1. 在0o C條件下，將NaNO2 的水溶液（1.4mL中含0.970g，14.0mmol）慢慢加入到4-溴-2-氯-3-氟苯胺（3.00g，13.4mmol）的4.0M HCl（14mL）溶液中。將混合物在室溫下攪拌1小時。在0o C條件下，加入(E)-N,N-二甲基-2-硝基乙烯-1-胺的乙腈溶液（15mL中含1.55g，13.4mmol）。反應混合物允許加熱至室溫，並攪拌3小時。將反應混合物用水（15mL）稀釋。對生成的懸浮液過濾。用冷的Et2 O（10mL）洗滌濾餅，得到橙色固體產物（3.50g，80.0%）。LC-MS (ESI) [M- H]- 323.8.1 H NMR (400 MHz, DMSO-d6 ): δ 13.70 (br s, 1H), 11.15 – 10.05 (m, 1 H), 7.80 (d, J = 6.4 Hz, 1H), 7.52 (d, J = 6.4 Hz, 1H)。Step 1. At 0 o C, slowly add an aqueous solution of NaNO 2 (0.970 g, 14.0 mmol in 1.4 mL) to 4-bromo-2-chloro-3-fluoroaniline (3.00 g, 13.4 mmol) 4.0M HCl (14mL) solution. The mixture was stirred at room temperature for 1 hour. Under conditions of 0 o C, was added (E) -N, N- dimethyl-2-nitroethylene 1-amine in acetonitrile (15mL containing 1.55g, 13.4mmol). The reaction mixture was allowed to warm to room temperature and stirred for 3 hours. The reaction mixture was diluted with water (15 mL). Filter the resulting suspension. The filter cake was washed with cold Et 2 O (10 mL) to obtain an orange solid product (3.50 g, 80.0%). LC-MS (ESI) [M- H] - 323.8 1 H NMR (400 MHz, DMSO-d 6):. Δ 13.70 (br s, 1H), 11.15 - 10.05 (m, 1 H), 7.80 (d, J = 6.4 Hz, 1H), 7.52 (d, J = 6.4 Hz, 1H).
步驟2. 在Ar下，(Z)-2((Z)-(4-溴-2-氯-3-氟苯基)二氮烯基)-2-硝基乙烯-1-醇（2.00g，6.16 mmol）的濃H2 SO4 （20mL）溶液在100o C條件下攪拌8小時。將反應混合物倒在冰上，然後用乙酸乙酯（30 mL X 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓條件下濃縮。利用矽膠柱色譜法（石油醚/乙酸乙酯=10/1）對殘餘物進行純化，得到黃色固體產物（400mg，21.2%）。LC-MS (ESI) [M+H]+ 308.1.1 H NMR (400 MHz, CDCl3 ): δ 8.77 (s, 1H), 8.36 (d, J = 6.4 Hz, 1H)。Step 2. Under Ar, (Z)-2((Z)-(4-bromo-2-chloro-3-fluorophenyl)diazenyl)-2-nitroethylene-1-ol (2.00g , 6.16 mmol) concentrated H 2 SO 4 (20 mL) solution was stirred at 100 o C for 8 hours. The reaction mixture was poured on ice, and then extracted with ethyl acetate (30 mL X 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=10/1) to obtain a yellow solid product (400 mg, 21.2%). LC-MS (ESI) [M+H] + 308.1. 1 H NMR (400 MHz, CDCl 3 ): δ 8.77 (s, 1H), 8.36 (d, J = 6.4 Hz, 1H).
步驟3. 將Fe（549mg，9.80mmol）、NH4 Cl（520mg，9.80mmol）加入到6-溴-8-氯-7-氟-3-硝基噌啉（600mg，1.96mmol）的EtOH/H2 O（7.5mL/2.5mL）溶液。生成的混合物在80o C條件下攪拌1小時。冷卻至室溫後，過濾混合物，濾液用H2 O（50mL）稀釋，並用EA（30 mL x 3）萃取。合併的有機相用Na2 SO4 乾燥，並在真空條件下濃縮得到殘餘物，並通過矽膠柱色譜純化（PE:EA = 20:1 to 2:1, v/v）得到為白色固體的標題化合物（500mg，產率93.1%）。LC-MS (ESI) [M+H]+ 278.1。Step 3. Add Fe (549mg, 9.80mmol), NH 4 Cl (520mg, 9.80mmol) to 6-bromo-8-chloro-7-fluoro-3-nitrocinnoline (600mg, 1.96mmol) in EtOH/ H 2 O (7.5mL/2.5mL) solution. The resulting mixture was stirred at 80 o C for 1 hour. After cooling to room temperature, the mixture was filtered, the filtrate was diluted with H 2 O (50 mL) and extracted with EA (30 mL x 3). The combined organic phase was dried with Na 2 SO 4 and concentrated under vacuum to obtain a residue, which was purified by silica gel column chromatography (PE:EA = 20:1 to 2:1, v/v) to obtain the title as a white solid Compound (500mg, yield 93.1%). LC-MS (ESI) [M+H] + 278.1.
根據通用步驟1，經由中間體B、環丙烷碳醯氯和4-甲基-3-(4,4,5,5-四甲基-1,3,2-二氧雜硼烷-2-基)吡啶（WO2018183964A1中已知的化合物）製備N-(8-氨基-7-氟-6-(4-甲基吡啶-3-基)噌啉-3-基)環丙烷甲醯胺。According to general procedure 1, via intermediate B, cyclopropane carbonyl chloride and 4-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2- Yl)pyridine (a compound known in WO2018183964A1) to prepare N-(8-amino-7-fluoro-6-(4-methylpyridin-3-yl)cinolin-3-yl)cyclopropanecarboxamide.
2-((8-2-((8- 氨基Amino -7--7- 氟fluorine -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-6-)-6- 異丙基Isopropyl -5,6--5,6- 二氫Dihydro -4H--4H- 吡唑並Pyrazolo [1,5- d][1,5- d] [1,4][1,4] 二氮雜卓Diazepam -7(8H)--7(8H)- 酮ketone (( 化合物Compound 24)twenty four)
反式Trans -N-(8--N-(8- 氨基Amino -7--7- 氟fluorine -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )-2-)-2- 氰基環丙烷Cyanocyclopropane -1--1- 甲醯胺Formamide (( 化合物Compound 25)25)
N-(8-N-(8- 氨基Amino -7--7- 氟fluorine -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )-2-)-2- 氰基環丁烷Cyanocyclobutane -1--1- 甲醯胺Formamide (( 化合物Compound 26)26)
2-(4-((8-2-(4-((8- 氨基Amino -7--7- 氟fluorine -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-1H-)-1H- 吡唑Pyrazole -1--1- 基base )) 丙腈Propionitrile (( 化合物Compound 27)27)
N-(8-N-(8- 氨基Amino -7--7- 氟fluorine -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 四氫Tetrahydro -2H--2H- 吡喃Pyran -2--2- 甲醯胺Formamide (( 化合物Compound 28)28)
N-(8-N-(8- 氨基Amino -7--7- 氟fluorine -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 四氫Tetrahydro -2H--2H- 吡喃Pyran -3--3- 甲醯胺Formamide (( 化合物Compound 29)29)
反式Trans -N-(8--N-(8- 氨基Amino -7--7- 氟fluorine -6-(8--6-(8- 甲基methyl -2,3--2,3- 二氫Dihydro -1H--1H- 吡啶並Pyrido [2,3-b] [1,4][2,3-b] [1,4] 惡嗪Oxazine -7--7- 基base )) 噌啉Cinolin --3---3- 基base )-2-)-2- 氰基環丙烷Cyanocyclopropane -1--1- 甲醯胺Formamide (( 化合物Compound 30)30)
2-((8-2-((8- 氨基Amino -7--7- 氟fluorine -6-(8--6-(8- 甲基methyl -2,3--2,3- 二氫Dihydro -1H--1H- 吡啶並Pyrido [2,3-B] [1,4][2,3-B] [1,4] 惡嗪Oxazine -7--7- 基base )) 噌啉Cinolin -3--3- 基base )))) 氨基Amino )-6-)-6- 異丙基Isopropyl -5,6--5,6- 二氫Dihydro -4H--4H- 吡唑並Pyrazolo [1,5-d] [1,4][1,5-d] [1,4] 二氮雜卓Diazepam -7(8H)--7(8H)- 酮ketone (( 化合物Compound 31)31)
N-(8-N-(8- 氨基Amino -7--7- 氟fluorine -6-(8--6-(8- 甲基methyl -2,3--2,3- 二氫Dihydro -1H--1H- 吡啶並Pyrido [2,3-b] [1,4][2,3-b] [1,4] 惡嗪Oxazine -7--7- 基base )) 噌啉Cinolin -3--3- 基base )-2-)-2- 氰基環丁烷Cyanocyclobutane -1--1- 甲醯胺Formamide (( 化合物Compound 32)32)
N-(8-N-(8- 氨基Amino -7--7- 氟fluorine -6-(8--6-(8- 甲基methyl -2,3--2,3- 二氫Dihydro -1H--1H- 吡啶並Pyrido [2,3-b] [1,4][2,3-b] [1,4] 惡嗪Oxazine -7--7- 基base )) 噌啉Cinolin -3--3- 基base ) -1-(1-) -1-(1- 甲基methyl -1H--1H- 吡唑Pyrazole -4--4- 基base )) 氮雜環丁烷Azetidine -2--2- 甲醯胺Formamide (( 化合物Compound 33)33)
2-((8-2-((8- 氨基Amino -7--7- 氟fluorine -6-(5--6-(5- 甲氧基Methoxy -4--4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-6-)-6- 異丙基Isopropyl -5,6--5,6- 二氫Dihydro -4H--4H- 吡唑並Pyrazolo [ 1,5-d] [1,4][1,5-d] [1,4] 二氮雜卓Diazepam -7(8H)--7(8H)- 酮ketone (( 化合物Compound 34)34)
(( 反式Trans )-N-(8-)-N-(8- 氨基Amino -7--7- 氟fluorine -6-(5--6-(5- 甲氧基Methoxy -4--4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )-2-)-2- 氰基環丙烷Cyanocyclopropane -1--1- 甲醯胺Formamide (( 化合物Compound 35)35)
6-(8-6-(8- 氨基Amino -7--7- 氟fluorine -3-((6--3-((6- 異丙基Isopropyl -7--7- 氧代Oxo -5,6,7,8--5,6,7,8- 四氫Tetrahydro -4H--4H- 吡唑並Pyrazolo [1,5-d] [1,4][1,5-d] [1,4] 二氮雜卓Diazepam - 2-- 2- 基base )) 氨基Amino )) 噌啉Cinolin -6--6- 基base )-5-)-5- 甲基苯並Methyl benzo [d][d] 惡唑Oxazole -2(3H)--2(3H)- 酮ketone (( 化合物Compound 36)36)
(( 反式Trans )-N-(8-)-N-(8- 氨基Amino -7--7- 氟fluorine -6-(5--6-(5- 甲基methyl -2--2- 氧代Oxo -2,3--2,3- 二氫苯並Dihydrobenzo [d][d] 惡唑Oxazole -6--6- 基base )) 噌啉Cinolin -3--3- 基base )-2-)-2- 氰基環丙烷Cyanocyclopropane -1--1- 甲醯胺Formamide (( 化合物Compound 37)37)
N-(8-N-(8- 氨基Amino -6-(5-(-6-(5-( 二甲基磷醯基Dimethylphosphoryl )-2-)-2- 甲基苯基Methyl phenyl )-7-)-7- 氟噌啉Flucinoline --3---3- 基base )-1-(1-)-1-(1- 甲基methyl -1H--1H- 吡唑Pyrazole -4--4- 基base )) 氮雜環丁烷Azetidine -2--2- 甲醯胺Formamide (( 化合物Compound 38)38)
N-(8-N-(8- 氨基Amino -6-(5-(-6-(5-( 二甲基磷醯基Dimethylphosphoryl )-2-)-2- 甲基苯基Methyl phenyl )-7-)-7- 氟噌啉Flucinoline -3--3- 基base )-2-)-2- 氰基環丁烷Cyanocyclobutane -1--1- 甲醯胺Formamide (( 化合物Compound 39)39)
(( 反式Trans )-N-(8-)-N-(8- 氨基Amino -6-(5-(-6-(5-( 二甲基磷醯基Dimethylphosphoryl )-2-)-2- 甲基苯基Methyl phenyl )-7-)-7- 氟噌啉Flucinoline -3--3- 基base )-2-)-2- 氰基環丙烷Cyanocyclopropane -1--1- 甲醯胺Formamide (( 化合物Compound 40)40)
2-((8-2-((8- 氨基Amino -6-(5-(-6-(5-( 二甲基磷醯基Dimethylphosphoryl )-2-)-2- 甲基苯基Methyl phenyl )-7-)-7- 氟噌啉Flucinoline -3--3- 基base )) 氨基Amino )-6-)-6- 異丙基Isopropyl -5,6--5,6- 二氫Dihydro -4H--4H- 吡唑並Pyrazolo [1,5-d] [1,4][1,5-d] [1,4] 二氮雜Diaza -7(8H)--7(8H)- 酮ketone (( 化合物Compound 41)41)
根據相關的脫氟類似物描述的步驟，製備化合物 24-41 。 Compounds 24-41 were prepared according to the procedures described in the relevant defluorinated analogs.
1-(2-((8- 氨基 -6-(4- 甲基吡啶 -3- 基 ) 噌啉 -3- 基 ) 氨基 )-7,8- 二氫 -1,6- 萘啶 -6(5H)- 基 ) 乙烷 -1- 酮 ( 化合物 52) 的製備 
步驟1. 根據通用步驟B 描述的方法，利用1-(2-氯-7,8-二氫-1,6-萘啶-6(5H)-基)乙酮，合成為黃色固體的1-(2-((8-氯-6-(4-甲基吡啶-3-基)噌啉-3-基)氨基)-7,8-二氫-1,6-萘啶-6(5H)-基)乙烯-1酮（60.0mg，24.3%）。LC-MS (ESI) [M+H]+ 445.1。Step 1. According to the method described in general step B , using 1-(2-chloro-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone, 1- (2-((8-Chloro-6-(4-methylpyridin-3-yl)cinoline-3-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H) -Base) ketene-1 (60.0 mg, 24.3%). LC-MS (ESI) [M+H] + 445.1.
步驟2. 根據通用步驟A 描述的方法（在微波下，150o C ，3h），合成棕色固體1-(2-((8-二苯亞甲基)氨基-6-(4-甲基吡啶-3-基)噌啉-3-基)氨基)-7,8-二氫-1,6-萘啶-6(5H)-基)乙烯-1酮（20.0mg，21.5%）。LC-MS (ESI) [M+H]+ 590.1。Step 2. According to the method described in general step A (under microwave, 150 o C, 3h), a brown solid 1-(2-((8-diphenylmethylene)amino-6-(4-methylpyridine) was synthesized -3-yl)Cinolin-3-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethen-1-one (20.0 mg, 21.5%). LC-MS (ESI) [M+H] + 590.1.
步驟3. 根據通用步驟C 描述的方法（室溫，1h），合成為黃色固體的化合物 52 （5.60mg，51.9%）。LC-MS (ESI) [M+H]+ 426.2.1 H NMR (400 MHz, DMSO-d6 ): δ 10.34 (d, J = 6.8 Hz, 1H), 8.70 (d, J = 1.6 Hz, 1H), 8.47 - 8.45 (m, 2H), 7.54 - 7.50 (m, 1H), 7.37 (d, J = 4.8 Hz, 1H), 7.26 - 7.22 (m, 1H), 6.89 - 6.87 (m, 1H), 6.61 - 6.60 (m, 3H), 4.60 (s, 1H), 4.55 (s, 1H), 3.78 - 3.74 (m, 2H), 2.98 - 2.82 (m, 2H), 2.32 (s, 3H), 2.10 (d, J = 4.4 Hz, 3H)。Step 3. According to the method described in general step C (room temperature, 1 h), compound 52 (5.60 mg, 51.9%) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 426.2. 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.34 (d, J = 6.8 Hz, 1H), 8.70 (d, J = 1.6 Hz, 1H ), 8.47-8.45 (m, 2H), 7.54-7.50 (m, 1H), 7.37 (d, J = 4.8 Hz, 1H), 7.26-7.22 (m, 1H), 6.89-6.87 (m, 1H), 6.61-6.60 (m, 3H), 4.60 (s, 1H), 4.55 (s, 1H), 3.78-3.74 (m, 2H), 2.98-2.82 (m, 2H), 2.32 (s, 3H), 2.10 ( d, J = 4.4 Hz, 3H).
6-(4- 甲基吡啶 -3- 基 )-N3-(6-( 甲基磺醯基 )-5,6,7,8- 四氫 -1,6- 萘啶 -2- 基 ) 噌啉 -3,8- 二胺 (53) 的製備 
2-2- 氯chlorine -6-(-6-( 甲基磺醯基Methylsulfonyl )-5,6,7,8-)-5,6,7,8- 四氫Tetrahydro -1,6--1,6- 萘啶的合成Synthesis of naphthyridine
在0o C條件下，將三乙胺（836mg，8.26mmol）和甲基磺醯氯（566mg，4.94mmol）加入到2-氯-5,6,7,8-四氫-1,6-萘啶二鹽酸鹽（400mg，1.66mmol）的N,N-二甲基甲醯胺（10 L）溶液中。將混合物在室溫下攪拌5小時。將混合物用水（100mL）稀釋，用乙酸乙酯（50 mL X 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾，在減壓條件下濃縮。利用矽膠柱色譜法（石油醚/乙酸乙酯=1/1）純化，得到為白色固體的2-氯-6-(甲基磺醯基)-5,6,7,8-四氫-1,6-萘啶（400mg，97.9%）。LC-MS (ESI) [M+H]+ 247.0.1 H NMR (400 MHz, DMSO-d6 ): δ 7.72 (d, J = 8.0 Hz, 1H), 7.37 (d, J = 8.0 Hz, 1H), 4.42 (s, 2H), 3.54 - 3.51 (m, 2H), 3.00 - 2.97 (m, 5H)。At 0 o C, add triethylamine (836mg, 8.26mmol) and methylsulfonyl chloride (566mg, 4.94mmol) to 2-chloro-5,6,7,8-tetrahydro-1,6- Naphthyridine dihydrochloride (400mg, 1.66mmol) in N,N-dimethylformamide (10 L) solution. The mixture was stirred at room temperature for 5 hours. The mixture was diluted with water (100 mL) and extracted with ethyl acetate (50 mL X 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purified by silica gel column chromatography (petroleum ether/ethyl acetate=1/1) to obtain 2-chloro-6-(methylsulfonyl)-5,6,7,8-tetrahydro-1 as a white solid ,6-Naphthyridine (400mg, 97.9%). LC-MS (ESI) [M+H] + 247.0. 1 H NMR (400 MHz, DMSO-d 6 ): δ 7.72 (d, J = 8.0 Hz, 1H), 7.37 (d, J = 8.0 Hz, 1H ), 4.42 (s, 2H), 3.54-3.51 (m, 2H), 3.00-2.97 (m, 5H).
根據化合物 52 相似的方法製備為黃色固體的化合物 53 （3.00mg，27.1%）。LC-MS (ESI) [M+H]+ 462.2.1 H NMR (400 MHz, DMSO-d6 ): δ 10.38 (s, 1H), 8.71 (s, 1H), 8.48 - 8.45 (m, 2H), 7.53 (d, J = 8.4 Hz, 1H), 7.37 (d, J = 4.4 Hz, 1H), 7.24 (d, J = 8.4 Hz, 1H), 6.89 (s, 1H), 6.62 - 6.60 (m, 3H), 4.33 (s, 2H), 3.54 - 3.51 (m, 2H), 3.00 - 2.97 (m, 5H), 2.32 (s, 3H)。 Compound 53 (3.00 mg, 27.1%) was prepared as a yellow solid according to a similar method to compound 52. LC-MS (ESI) [M+H] + 462.2. 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.38 (s, 1H), 8.71 (s, 1H), 8.48-8.45 (m, 2H) , 7.53 (d, J = 8.4 Hz, 1H), 7.37 (d, J = 4.4 Hz, 1H), 7.24 (d, J = 8.4 Hz, 1H), 6.89 (s, 1H), 6.62-6.60 (m, 3H), 4.33 (s, 2H), 3.54-3.51 (m, 2H), 3.00-2.97 (m, 5H), 2.32 (s, 3H).
2-((8- 氨基 -6-(4- 甲基吡啶 -3- 基 ) 噌啉 -3- 基 ) 氨基 )-6- 異丙基 -5,8- 二氫 -1,6- 萘啶 -7(6H)- 酮 (56) 的製備 
步驟1. 在室溫下，將NaBH3 CN (630 mg, 10.0 mmol)慢慢加入到2,6-二氯吡啶-3-甲醛（700mg，4.00mmol）、丙基-2-胺（281 mg, 4.80 mmol）和Ti(i-PrO)4 (400 mg, 1.40 mmol)的甲醇（10mL）溶液。反應混合物在25o C下攪拌過夜，並在減壓條件下濃縮。殘餘物利用矽膠柱色譜法純化，並用石油醚/乙酸乙酯（10:1 ~ 8:1）洗脫得到為無色液體的產物（780 mg, 89.0%）。LC-MS (ESI) [M+H]+ 219.1.1 H NMR (400 MHz, CDCl3 ): δ 7.99 (d, J = 8.0 Hz, 1H), 7.28 (d, J = 8.0 Hz, 1H), 3.95 (s, 2H), 2.95 - 2.92 (m, 1 H), 1.20 (d, J = 6.4 Hz, 6H)。Step 1. At room temperature, slowly add NaBH 3 CN (630 mg, 10.0 mmol) to 2,6-dichloropyridine-3-carbaldehyde (700 mg, 4.00 mmol), propyl-2-amine (281 mg , 4.80 mmol) and Ti(i-PrO) 4 (400 mg, 1.40 mmol) in methanol (10 mL). The reaction mixture was stirred overnight at 25 o C, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with petroleum ether/ethyl acetate (10:1 ~ 8:1) to obtain the product (780 mg, 89.0%) as a colorless liquid. LC-MS (ESI) [M+H] + 219.1. 1 H NMR (400 MHz, CDCl 3 ): δ 7.99 (d, J = 8.0 Hz, 1H), 7.28 (d, J = 8.0 Hz, 1H), 3.95 (s, 2H), 2.95-2.92 (m, 1 H), 1.20 (d, J = 6.4 Hz, 6H).
步驟2. 在0o C下，將HATU（1.92 g, 5.04 mmol）加入到N-((2,6-二氯吡啶-3-基)甲基)丙基-2-胺（850mg，3.88mmol）、3-(叔丁氧基)-3-丙酮酸（622 mg, 3.88 mmol）和DIPEA（1.00 g, 7.76 mmol）的DMF（9mL）溶液中。將混合物在室溫下攪拌1小時。反應混合物用水（20mL）稀釋，用乙酸乙酯（20 mL X 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓條件下濃縮。利用矽膠柱色譜法（石油醚/乙酸乙酯=10/1）純化得到白色固體產物（1.20g，90.7%）。LC-MS (ESI) [M-56+H]+ 305.3.1 H NMR (400 MHz, MeOD-d4 ): δ 7.77 (d, J = 8.0 Hz, 1H), 7.38 (d, J = 8.0 Hz, 1H), 4.50 (s, 2H), 4.23 – 4.21 (m, 1 H), 3.64 (s, 2H), 1.51 (s, 9H), 1.19 (d, J = 6.4 Hz, 1H)。Step 2. Add HATU (1.92 g, 5.04 mmol) to N-((2,6-dichloropyridin-3-yl)methyl)propyl-2-amine (850 mg, 3.88 mmol) at 0 o C ), 3-(tert-butoxy)-3-pyruvate (622 mg, 3.88 mmol) and DIPEA (1.00 g, 7.76 mmol) in DMF (9mL). The mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL X 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Purified by silica gel column chromatography (petroleum ether/ethyl acetate=10/1) to obtain a white solid product (1.20g, 90.7%). LC-MS (ESI) [M-56+H] + 305.3. 1 H NMR (400 MHz, MeOD-d 4 ): δ 7.77 (d, J = 8.0 Hz, 1H), 7.38 (d, J = 8.0 Hz , 1H), 4.50 (s, 2H), 4.23 – 4.21 (m, 1 H), 3.64 (s, 2H), 1.51 (s, 9H), 1.19 (d, J = 6.4 Hz, 1H).
步驟3. 在Ar條件下，將NaH（222mg，5.54mmol）慢慢地加入到3-(((2,6-二氯吡啶-3-基)甲基)(異丙基)氨基)-3-氧代丙酸叔丁基酯（1.00g，2.77mmol）的DMF（20mL）溶液中。生成的混合物在70o C條件下攪拌5小時。將反應混合物用NH4 Cl水溶液（30mL）稀釋，用乙酸乙酯（30 mL X 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓條件下濃縮。利用矽膠柱色譜法（石油醚/乙酸乙酯=5/1）純化得到為黃色油狀物的產物（300mg，33.0%）。LC-MS (ESI) [M+H]+ 325.1.1 H NMR (400 MHz, CDCl3 ): δ 7.53 (d, J = 8.0 Hz, 1H), 7.27 (d, J = 8.0 Hz, 1H), 5.00 - 4.97 (m, 1 H), 4.66 (s, 1H), 4.55 (d, J = 15.6 Hz, 1H), 4.23 (d, J = 15.6 Hz, 1H), 1.51 (s, 9H), 1.22 (dd, J = 15.6 Hz, 6.4 Hz, 6H)。Step 3. Under Ar conditions, slowly add NaH (222mg, 5.54mmol) to 3-(((2,6-dichloropyridin-3-yl)methyl)(isopropyl)amino)-3 -Tert-butyl oxopropionate (1.00 g, 2.77 mmol) in DMF (20 mL). The resulting mixture was stirred at 70 o C for 5 hours. The reaction mixture was diluted with NH 4 Cl aqueous solution (30 mL) and extracted with ethyl acetate (30 mL×3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Purified by silica gel column chromatography (petroleum ether/ethyl acetate=5/1), the product (300 mg, 33.0%) was obtained as a yellow oil. LC-MS (ESI) [M+H] + 325.1. 1 H NMR (400 MHz, CDCl 3 ): δ 7.53 (d, J = 8.0 Hz, 1H), 7.27 (d, J = 8.0 Hz, 1H), 5.00-4.97 (m, 1 H), 4.66 (s, 1H), 4.55 (d, J = 15.6 Hz, 1H), 4.23 (d, J = 15.6 Hz, 1H), 1.51 (s, 9H), 1.22 ( dd, J = 15.6 Hz, 6.4 Hz, 6H).
步驟4. 在0℃條件下將TFA（0.6mL）加入到2-氯-6-異丙基-7-氧代-5,6,7,8-四氫-1,6-萘啶-8-羧酸叔丁基酯（300mg，0.924mmol）的二氯甲烷（3mL）溶液中。將混合物在室溫下攪拌2小時。將混合物在減壓條件下濃縮。殘餘物用二氯甲烷（15mL）溶解，並用NaHCO3 水溶液（15 mL X 3）洗滌。將有機層用鹽水洗滌，並無水硫酸鈉乾燥，過濾並在減壓條件下濃縮。將粗產物（180mg，87.0%）直接用於下一步驟。LC-MS (ESI) [M+H]+ 225.1。Step 4. Add TFA (0.6 mL) to 2-chloro-6-isopropyl-7-oxo-5,6,7,8-tetrahydro-1,6-naphthyridine-8 at 0℃ -Tert-butyl carboxylate (300 mg, 0.924 mmol) in dichloromethane (3 mL). The mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure. The residue was dissolved with dichloromethane (15 mL) and washed with aqueous NaHCO 3 (15 mL X 3). The organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product (180 mg, 87.0%) was used directly in the next step. LC-MS (ESI) [M+H] + 225.1.
採用與化合物 52 相似的方法製備為黃色固體的化合物 56 （1.63mg，44.0%）。LC-MS (ESI) [M+H]+ 440.2.1 H NMR (400 MHz, MeOD-d4 ): δ 8.76 (s, 1H), 8.42 – 8.41 (m, 2H), 7.61 (d, J = 8.4 Hz, 1H), 7.40 (d, J = 5.2 Hz, 1H), 7.14 (d, J = 8.4 Hz, 1H), 6.95 (s, 1H), 6.70 (s, 1H), 4.94 – 4.92 (m, 1H), 4.44 (s, 2H), 3.74 (s, 2H), 2.40 (s, 3H), 1.24 (d, J = 6.8 Hz, 6H)。 Compound 56 (1.63 mg, 44.0%) was prepared as a yellow solid using a method similar to that of compound 52. LC-MS (ESI) [M+H] + 440.2. 1 H NMR (400 MHz, MeOD-d 4 ): δ 8.76 (s, 1H), 8.42 – 8.41 (m, 2H), 7.61 (d, J = 8.4 Hz, 1H), 7.40 (d, J = 5.2 Hz, 1H), 7.14 (d, J = 8.4 Hz, 1H), 6.95 (s, 1H), 6.70 (s, 1H), 4.94 – 4.92 (m, 1H), 4.44 (s, 2H), 3.74 (s, 2H), 2.40 (s, 3H), 1.24 (d, J = 6.8 Hz, 6H).
在圖表1中，根據通用步驟製備以下化合物。In Chart 1, the following compounds were prepared according to the general procedure.
2-((8-2-((8- 氨基Amino -7--7- 氟fluorine -6-(5--6-(5- 甲基methyl -1H--1H- 咪唑Imidazole -1--1- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-6-)-6- 異丙基Isopropyl -5,6--5,6- 二氫Dihydro -4H--4H- 吡唑並Pyrazolo [1,5-d][1,4][1,5-d][1,4] 二氮雜卓Diazepam -7(8H)--7(8H)- 酮ketone (( 化合物Compound 42)42) ；；
2-((8-2-((8- 氨基Amino -7--7- 氟fluorine -6-(-6-( 噻唑Thiazole -5--5- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-6-)-6- 異丙基Isopropyl -5,6--5,6- 二氫Dihydro -4H--4H- 吡唑並Pyrazolo [1,5-d][1,4][1,5-d][1,4] 二氮雜卓Diazepam -7(8H)--7(8H)- 酮ketone (( 化合物Compound 43)43) ；；
2-((8-2-((8- 氨基Amino -6-(5--6-(5- 甲基methyl -1H--1H- 咪唑Imidazole -1--1- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-6-)-6- 異丙基Isopropyl -5,6--5,6- 二氫Dihydro -4H--4H- 吡唑並Pyrazolo [1,5-d][1,4][1,5-d][1,4] 二氮雜卓Diazepam -7(8H)--7(8H)- 酮ketone (( 化合物Compound 44)44) ；；
2-((8-2-((8- 氨基Amino -6-(-6-( 噻唑Thiazole -5--5- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-6-)-6- 異丙基Isopropyl -5,6--5,6- 二氫Dihydro -4H--4H- 吡唑並Pyrazolo [1,5-d] [1,4][1,5-d] [1,4] 二氮雜卓Diazepam -7(8H)--7(8H)- 酮ketone (( 化合物Compound 45)45) ；；
2-((8-2-((8- 氨基Amino -7--7- 氟fluorine -6-(1--6-(1- 甲基methyl -1H--1H- 咪唑Imidazole -5--5- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-6-)-6- 異丙基Isopropyl -5,6--5,6- 二氫Dihydro -4H--4H- 吡唑並Pyrazolo [1,5-d][1,4][1,5-d][1,4] 二氮雜卓Diazepam -7(8H)--7(8H)- 酮ketone (( 化合物Compound 46)46) ；；
2-((8-2-((8- 氨基Amino -6-(1--6-(1- 乙基Ethyl -1H--1H- 咪唑Imidazole -5--5- 基base )-7-)-7- 氟噌啉Flucinoline -3--3- 基base )) 氨基Amino )-6-)-6- 異丙基Isopropyl -5,6--5,6- 二氫Dihydro -4H--4H- 吡唑並Pyrazolo [1,5-d][1,4][1,5-d][1,4] 二氮雜卓Diazepam -7(8H)--7(8H)- 酮ketone (( 化合物Compound 47)47) ；；
2-((8-2-((8- 氨基Amino -6-(1--6-(1- 甲基methyl -1H--1H- 咪唑Imidazole -5--5- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-6-)-6- 異丙基Isopropyl -5,6--5,6- 二氫Dihydro -4H--4H- 吡唑並Pyrazolo [1,5-d][1,4][1,5-d][1,4] 二氮雜卓Diazepam -7(8H)--7(8H)- 酮ketone (( 化合物Compound 48)48) ；；
2-((8-2-((8- 氨基Amino -6-(1--6-(1- 乙基Ethyl -1H--1H- 咪唑Imidazole -5--5- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-6-)-6- 異丙基Isopropyl -5,6--5,6- 二氫Dihydro -4H--4H- 吡唑並Pyrazolo [1,5-d][1,4][1,5-d][1,4] 二氮雜卓Diazepam -7(8H)--7(8H)- 酮ketone (( 化合物Compound 49)49) ；；
1-(2-((8-1-(2-((8- 氨基Amino -7--7- 氟fluorine -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-7,8-)-7,8- 二氫Dihydro -1,6--1,6- 萘啶Naphthyridine -6(5H)--6(5H)- 基base )) 乙烷Ethane -1--1- 酮ketone (( 化合物Compound 50)50) ；；
7-7- 氟fluorine -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )-N3-(6-()-N3-(6-( 甲基磺醯基Methylsulfonyl )-5,6,7,8-)-5,6,7,8- 四氫Tetrahydro -1,6--1,6- 萘啶Naphthyridine -2--2- 基base )) 噌啉Cinolin -3,8--3,8- 二胺Diamine (( 化合物Compound 51)51) ；；
2-((8-2-((8- 氨基Amino -7--7- 氟fluorine -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-6-)-6- 異丙基Isopropyl -5,8--5,8- 二氫Dihydro -1,6--1,6- 萘啶Naphthyridine -7(6H)--7(6H)- 酮ketone (( 化合物Compound 54)54) ；；
2-((8-2-((8- 氨基Amino -7--7- 氟fluorine -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-6-)-6- 異丙基Isopropyl -7,8--7,8- 二氫Dihydro -1,6--1,6- 萘啶Naphthyridine -5(6H)--5(6H)- 酮ketone (( 化合物Compound 55)55) ；；
2-((8- 氨基 -6-(4- 甲基吡啶 -3- 基 ) 噌啉 -3- 基 ) 氨基 )-6- 異丙基 -7,8- 二氫 -1,6- 萘啶 -5(6H)- 酮 ( 化合物 57) ； 
在圖表2中，根據通用步驟製備以下化合物。In Figure 2, the following compounds were prepared according to the general procedure.
7-7- 氟fluorine -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )-N3-(5-)-N3-(5- 嗎啉代吡啶Morpholinopyridine -2--2- 基base )) 噌啉Cinolin -3,8--3,8- 二胺Diamine (( 化合物Compound 58);58);
7-7- 氟fluorine -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )-N3-(5-)-N3-(5- 嗎啉代吡嗪Morpholinopyrazine -2--2- 基base )) 噌啉Cinolin -3,8--3,8- 二胺Diamine (( 化合物Compound 59);59);
6-(4-6-(4- 甲基吡啶Picoline -3--3- 基base )-N3-(6-)-N3-(6- 嗎啉代噠嗪Morpholinopyridazine -3--3- 基base )) 噌啉Cinolin -3,8--3,8- 二胺Diamine (( 化合物Compound 60);60);
6-(4-6-(4- 甲基吡啶Picoline -3--3- 基base )-N3-(5-(4-()-N3-(5-(4-( 甲基磺醯基Methylsulfonyl )) 呱嗪Pyrazine -1--1- 基base )) 吡啶Pyridine -2--2- 基base )) 噌啉Cinolin -3,8--3,8- 二胺Diamine (( 化合物Compound 61);61);
1-(4-(6-((8-1-(4-(6-((8- 氨基Amino -7--7- 氟fluorine -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )) 吡啶Pyridine -3--3- 基base )) 呱嗪Pyrazine -1--1- 基base )) 乙烷Ethane -1--1- 酮ketone (( 化合物Compound 62);62);
4-(6-((8-4-(6-((8- 氨基Amino -7--7- 氟fluorine -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )) 吡啶Pyridine -3--3- 基base )) 呱嗪Pyrazine -2--2- 酮ketone (( 化合物Compound 63);63);
4-(6-((8-4-(6-((8- 氨基Amino -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )) 噠嗪Pyridazine -3--3- 基base )) 硫代嗎啉Thiomorpholine 1,1-1,1- 二氧化物Dioxide (( 化合物Compound 64);64);
4-(6-((8-4-(6-((8- 氨基Amino -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )) 吡啶Pyridine -3--3- 基base )) 硫代嗎啉Thiomorpholine 1,1-1,1- 二氧化物Dioxide (( 化合物Compound 65);65);
1-(6-((8-1-(6-((8- 氨基Amino -7--7- 氟fluorine -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )) 吡啶Pyridine -3--3- 基base )) 吡咯烷Pyrrolidine -2--2- 酮ketone (( 化合物Compound 66);66);
1-(6-((8-1-(6-((8- 氨基Amino -7--7- 氟fluorine -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )) 吡啶Pyridine -3--3- 基base )) 呱啶Pyridine -2--2- 酮ketone (( 化合物Compound 67);67);
1-(6-((8- 氨基 -7- 氟 -6-(4- 甲基吡啶 -3- 基 ) 噌啉 -3- 基 ) 氨基 ) 吡啶 -3- 基 ) 氮雜環丁烷 -2- 酮 ( 化合物 68). 
N-(8- 氨基 -6-(4- 甲基吡啶 -3- 基 ) 吡啶並 [3,4-c] 噠嗪 -3- 基 ) 環丙烷甲醯胺（ 69 ）的製備 
步驟1. 在0o C條件下，將NaH（4.83g，121mmol）加入到丙二酸叔丁基甲酯（25g，133mmol）的DMF（250mL）的溶液中。在0o C條件下攪拌1小時後，在0o C條件下加入4,6-二氯噠嗪-3-羧酸甲酯（25g，121mmol）。將混合物在室溫攪拌18小時，然後用NH4 Cl水溶液（1L）稀釋，用乙酸乙酯（200 mL * 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓條件下濃縮。利用柱色譜法（PE/EA=3/1）純化得到為黃色油的2-(6-氯-3-(甲氧羰基))噠嗪-4-基)丙二酸1-(叔丁基)3-乙基酯（25.0g，43.2%產率）。LC-MS (ESI) [M+H]+ 359.2。Step 1. At 0 o C, add NaH (4.83g, 121mmol) to a solution of tert-butyl methyl malonate (25g, 133mmol) in DMF (250mL). After stirring at 0 o C for 1 hour, 4,6-dichloro-pyridazine-3-carboxylate (25g, 121mmol) at 0 o C condition. The mixture was stirred at room temperature for 18 hours, then diluted with aqueous NH 4 Cl (1 L), and extracted with ethyl acetate (200 mL * 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Purified by column chromatography (PE/EA=3/1) to obtain 2-(6-chloro-3-(methoxycarbonyl))pyridazin-4-yl)malonic acid 1-(tert-butyl) as yellow oil ) 3-Ethyl ester (25.0 g, 43.2% yield). LC-MS (ESI) [M+H] + 359.2.
步驟2. 在0o C條件下，將TFA（50.0mL）加入到2-(6-氯-3-(甲氧羰基))噠嗪-4-基)丙二酸1-(叔丁基)3-乙基酯（20.0g，55.7mmol）的DCM（100mL）溶液中。在室溫下攪拌1.5小時後，利用NaHCO3水溶液調整混合物的pH=7，並用DCM（100 mL * 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，並在減壓條件下濃縮。利用柱色譜法（PE/EA=2/1）對殘餘物進行純化，得到為白色固體的6-氯-4-(2-乙氧基-2-氧代乙基)噠嗪-3-羧酸甲酯（13.7g，95.1%產率）。LC-MS (ESI) [M+H]+ 258.8.1 H NMR (400 MHz, CDCl3 ): δ 7.52 (s, 1H), 4.19 (q, J = 6.8 Hz, 2H), 4.04 (s, 3H), 4.02 (s, 2H), 1.27 (t, J = 6.8 Hz, 3H)。Step 2. Add TFA (50.0 mL) to 2-(6-chloro-3-(methoxycarbonyl))pyridazin-4-yl)malonic acid 1-(tert-butyl) at 0 o C 3-ethyl ester (20.0 g, 55.7 mmol) in DCM (100 mL). After stirring at room temperature for 1.5 hours, the pH of the mixture was adjusted to 7 with an aqueous NaHCO3 solution, and extracted with DCM (100 mL * 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (PE/EA=2/1) to obtain 6-chloro-4-(2-ethoxy-2-oxoethyl)pyridazine-3-carboxylate as a white solid Methyl acid methyl ester (13.7 g, 95.1% yield). LC-MS (ESI) [M+H] + 258.8. 1 H NMR (400 MHz, CDCl 3 ): δ 7.52 (s, 1H), 4.19 (q, J = 6.8 Hz, 2H), 4.04 (s, 3H ), 4.02 (s, 2H), 1.27 (t, J = 6.8 Hz, 3H).
步驟3. 在密封管中，將6-氯-4-(2-乙氧基-2-氧代乙基)噠嗪-3-羧酸甲酯（12g，46.4mmol）的NH3 /MeOH（200mL，7mol/L）溶液在100℃條件下攪拌18小時。混合物冷卻至室溫，並濃縮得到紅色固體3-氯吡啶並[3,4-c]噠嗪-6,8-二醇（8.5g，92.8%產率）。LC-MS (ESI) [M+H]+ 198.0.1 H NMR (400 MHz, DMSO-d4 ): δ 10.06 (s, 1H), 7.10 (s, 4H), 6.75 (s, 1H), 4.60 (s, 1H)。Step 3. In a sealed tube, mix 6-chloro-4-(2-ethoxy-2-oxoethyl)pyridazine-3-carboxylic acid methyl ester (12g, 46.4mmol) in NH 3 /MeOH ( 200mL, 7mol/L) The solution was stirred at 100°C for 18 hours. The mixture was cooled to room temperature and concentrated to obtain 3-chloropyrido[3,4-c]pyridazine-6,8-diol (8.5 g, 92.8% yield) as a red solid. LC-MS (ESI) [M+H] + 198.0. 1 H NMR (400 MHz, DMSO-d 4 ): δ 10.06 (s, 1H), 7.10 (s, 4H), 6.75 (s, 1H), 4.60 (s, 1H).
步驟4. 在室溫條件下，將POBr3 （7.30g，25.4mmol）一次性加入到DIEA（1.31g，10.2mmol）的MeCN（30mL）溶液中，然後將3-氯吡啶並[3,4-c]噠嗪-6,8-二醇（1.00g，5.08mmol）一次性加入。反應在90℃下攪拌1小時。將冷的混合物倒在冰水（100mL）上，並用乙酸乙酯（100 mL * 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓條件下濃縮。利用柱色譜法對殘餘物進行純化，用石油醚/乙酸乙酯（50:1 ~ 40:1）洗脫得到黃色固體產物（400mg，24.3%產率）。LC-MS (ESI) [M+H]+ 323.8。Step 4. At room temperature, add POBr 3 (7.30g, 25.4mmol) to DIEA (1.31g, 10.2mmol) in MeCN (30mL) at a time, and then add 3-chloropyrido[3,4 -c] Pyridazine-6,8-diol (1.00 g, 5.08 mmol) was added all at once. The reaction was stirred at 90°C for 1 hour. The cold mixture was poured on ice water (100 mL) and extracted with ethyl acetate (100 mL * 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography and eluted with petroleum ether/ethyl acetate (50:1-40:1) to obtain a yellow solid product (400mg, 24.3% yield). LC-MS (ESI) [M+H] + 323.8.
步驟5.在0℃條件下，將6,8-二溴-3-氯吡啶並[3,4-c]噠嗪（1.20g，3.72mmol）的THF（20mL）溶液慢慢地加入到NH3 .H2 O (10 mL)中。反應混合物在室溫下攪拌16小時。將混合物濃縮，去除溶劑THF。利用MeOH（6mL）稀釋生成的懸浮液。然後將混合物過濾，並乾燥得到黃色固體產物（650mg，57.0%產率）。LC-MS (ESI) [M+H]+ 304.8。Step 5. At 0℃, add 6,8-dibromo-3-chloropyrido[3,4-c]pyridazine (1.20g, 3.72mmol) in THF (20mL) solution slowly to NH 3. In H 2 O (10 mL). The reaction mixture was stirred at room temperature for 16 hours. The mixture was concentrated and the solvent THF was removed. The resulting suspension was diluted with MeOH (6 mL). The mixture was then filtered and dried to obtain a yellow solid product (650 mg, 57.0% yield). LC-MS (ESI) [M+H] + 304.8.
步驟6. 將在NMP（2mL）中的6,8-二溴吡啶並[3,4-c]噠嗪-3-胺（650mg，2.14mmol）和雙-2,4-二甲氧基苄基胺（2.70g，8.55mmol）的混合物在120℃下攪拌16小時。將冷的混合物倒在冰水（10mL）上，並用乙酸乙酯（20 mL * 3）萃取。合併的有機層利用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓條件下濃縮。利用快速柱色譜法對殘餘物進行純化，並用石油醚/乙酸乙酯（65%:35%）洗脫得到黃色固體產物（140mg，12.1%產率）。LC-MS (ESI) [M+H]+ 541.9。Step 6. Combine 6,8-dibromopyrido[3,4-c]pyridazin-3-amine (650mg, 2.14mmol) and bis-2,4-dimethoxybenzyl in NMP (2mL) A mixture of base amines (2.70 g, 8.55 mmol) was stirred at 120°C for 16 hours. The cold mixture was poured onto ice water (10 mL) and extracted with ethyl acetate (20 mL * 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography and eluted with petroleum ether/ethyl acetate (65%:35%) to obtain a yellow solid product (140 mg, 12.1% yield). LC-MS (ESI) [M+H] + 541.9.
步驟7. 在0℃條件下，將環丙烷碳醯氯（34.0mg，0.325mmol）慢慢地加入到6-溴-N8,N8-雙(2,4-二甲氧基苄基)吡啶並[3,4-c]噠嗪-3,8-二胺（70.0mg，0.130mmol）和Et3N（53.0mg，0.520mmol）的DCM（3mL）溶液中。生成的混合物在室溫下攪拌過夜。混合物用NaHCO3 水溶液（5mL）稀釋，用乙酸乙酯（5 mL * 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓條件下濃縮。利用prep-TLC對殘餘物進行純化得到為黃色油狀物的產物（75.0mg，85.0%產率）。Step 7. At 0°C, slowly add cyclopropanecarbon chloride (34.0mg, 0.325mmol) to 6-bromo-N8,N8-bis(2,4-dimethoxybenzyl)pyrido [3,4-c] Pyridazine-3,8-diamine (70.0 mg, 0.130 mmol) and Et3N (53.0 mg, 0.520 mmol) in DCM (3 mL). The resulting mixture was stirred overnight at room temperature. The mixture was diluted with aqueous NaHCO 3 (5 mL) and extracted with ethyl acetate (5 mL * 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by prep-TLC to obtain the product as a yellow oil (75.0 mg, 85.0% yield).
步驟8. 根據通用步驟F描述的方法，合成為黃色固體的標題產物（14.0mg，20.0%產率）。LC-MS (ESI) [M+H]+ 621.0。Step 8. According to the method described in General Procedure F, the title product (14.0 mg, 20.0% yield) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 621.0.
步驟9.將N-(8-(雙(2,4-二甲氧基苄基)氨基)-6-(4-甲基吡啶-3-基)吡啶並[3,4-c]噠嗪-3-基)環丙烷甲醯胺（14.0mg，0.0226mmol）的TFA（0.5mL）溶液在50℃條件下攪拌3小時。將反應混合物用氬氣清除，去除TFA。利用prep-HPLC對粗產物進行純化得到黃色固體產物（6.75mg，93.3%產率）。LC-MS (ESI) [M+H]+ 321.0.1 H NMR (400 MHz, MeOD-d4 ): δ 8.60 (s, 1H), 8.52 (d, J = 5.2 Hz, 1H), 7.69 (s, 1H), 7.48 (d, J = 5.2 Hz, 1H), 6.24 (s, 1H), 2.44 (s, 3H), 2.43 – 2.42 (m, 1H), 1.13 – 1.11 (m, 2H), 1.04 – 1.01 (m, 2H)。Step 9. Add N-(8-(bis(2,4-dimethoxybenzyl)amino)-6-(4-methylpyridin-3-yl)pyrido[3,4-c]pyridazine A solution of -3-yl)cyclopropanecarboxamide (14.0mg, 0.0226mmol) in TFA (0.5mL) was stirred at 50°C for 3 hours. The reaction mixture was purged with argon to remove TFA. The crude product was purified by prep-HPLC to obtain a yellow solid product (6.75 mg, 93.3% yield). LC-MS (ESI) [M+H] + 321.0. 1 H NMR (400 MHz, MeOD-d 4 ): δ 8.60 (s, 1H), 8.52 (d, J = 5.2 Hz, 1H), 7.69 (s , 1H), 7.48 (d, J = 5.2 Hz, 1H), 6.24 (s, 1H), 2.44 (s, 3H), 2.43 – 2.42 (m, 1H), 1.13 – 1.11 (m, 2H), 1.04 – 1.01 (m, 2H).
( 反式 )-N-(8- 氨基 -6-(5- 氨基 -4- 甲基吡啶 -3- 基 ) 噌啉 -3- 基 )-2-(1- 甲基 -1H- 吡唑 -4 基 ) 環丙烷 -1- 甲醯胺（ 70 ）的製備 
(( 叔丁氧基羰基Tert-Butoxycarbonyl )(5-(8-)(5-(8- 氯chlorine -3--3- 氨基噌啉Amino Cinoline -6--6- 基base )-4-)-4- 甲基吡啶Picoline -3--3- 基base )) 氨基甲酸叔丁基酯Tert-butyl carbamate （( 中間體Intermediate FF ）) 的製備Preparation
根據通用步驟D描述的方法（90℃ 2h，利用中間體A和(叔丁氧基羰基)(4-甲基-5-(4,4,5,5-四甲基-1,3,2-二氧雜硼烷-2-基)吡啶-3-基)氨基甲酸叔丁基酯），合成為無色油狀物的標題產物（中間體F）（930mg，41.3%）。LC-MS (ESI) [M+H]+ 486.2。According to the method described in general procedure D (90℃ 2h, using intermediate A and (tert-butoxycarbonyl)(4-methyl-5-(4,4,5,5-tetramethyl-1,3,2) -Dioxaborane-2-yl)pyridin-3-yl)tert-butyl carbamate), the title product (Intermediate F) (930 mg, 41.3%) was synthesized as a colorless oil. LC-MS (ESI) [M+H] + 486.2.
步驟1. 在0℃條件下，將N,N-二甲基甲醯胺（1滴）和草醯氯（251mg，2.11mmol）加入到反式-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-羧酸（70mg，0.422mol）的二氯甲烷（5.00mL）溶液中。將混合物在室溫攪拌2小時。將混合物在減壓條件下濃縮。將殘餘物溶解在二氯甲烷（5.00mL）中，並加入到(5-(3-氨基-8-氯噌啉-6-基)-4-甲基吡啶-3-基)(叔丁氧基羰基)氨基甲酸叔丁基酯（137mg，0.282mmol）和三乙胺（128mg，1.27mmol）的DCM（5.00mL）溶液中。將混合物在室溫下攪拌2小時。將混合物用水（20mL）稀釋，用二氯甲烷（20 mL X 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓條件下濃縮。將殘餘物用矽膠柱色譜法（二氯甲烷/甲醇=20/1）純化，得到為黃色固體的(叔丁氧基羰基)-(5-(8-氯-3-(反式-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯氨基)噌啉-6-基)-4-甲基吡啶-3-基)氨基甲酸叔丁基酯（130mg，72.6%）。LC-MS (ESI) [M+H]+ 634.3。Step 1. At 0℃, add N,N-dimethylformamide (1 drop) and oxalic chloride (251mg, 2.11mmol) to trans-2-(1-methyl-1H-pyridine). (Azol-4-yl)cyclopropane-1-carboxylic acid (70 mg, 0.422 mol) in dichloromethane (5.00 mL). The mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure. The residue was dissolved in dichloromethane (5.00 mL) and added to (5-(3-amino-8-chlorocinoline-6-yl)-4-methylpyridin-3-yl)(tert-butoxy Benzylcarbonyl) carbamate (137mg, 0.282mmol) and triethylamine (128mg, 1.27mmol) in DCM (5.00mL). The mixture was stirred at room temperature for 2 hours. The mixture was diluted with water (20 mL) and extracted with dichloromethane (20 mL X 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane/methanol=20/1) to obtain (tert-butoxycarbonyl)-(5-(8-chloro-3-(trans-2- (1-Methyl-1H-pyrazol-4-yl)cyclopropane-1-carboxamide)cinoline-6-yl)-4-methylpyridin-3-yl)tert-butyl carbamate (130mg , 72.6%). LC-MS (ESI) [M+H] + 634.3.
步驟2. 根據通用步驟A 描述的方法，合成為黃色固體的(叔丁氧基羰基)(5-(8-((二苯亞甲基)氨基)-3-(-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺基)噌啉-6-基)-4-甲基吡啶-3-基)氨基甲酸叔丁基酯（45mg，21.5%）。LC-MS (ESI) [M+H]+ 779.4。Step 2. According to the method described in general step A , (tert-butoxycarbonyl)(5-(8-((diphenylmethylene)amino)-3-(-2-(1-methyl) was synthesized as a yellow solid -1H-pyrazol-4-yl) cyclopropane-1-carboxamido) cinnolin-6-yl)-4-methylpyridin-3-yl) tert-butyl carbamate (45mg, 21.5% ). LC-MS (ESI) [M+H] + 779.4.
步驟3. 根據通用步驟C 描述的方法，合成為黃色固體的(5-(8-氨基-3-(-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺基)噌啉-6-基)-4-甲基吡啶-3-基)( 叔丁氧基羰基)氨基甲酸叔丁基酯（30mg，96.8%）。LC-MS (ESI) [M+H]+ 615.3。Step 3. According to the method described in general step C , (5-(8-amino-3-(-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1- Carboxamido) cinnolin-6-yl)-4-methylpyridin-3-yl) (tert-butoxycarbonyl) tert-butyl carbamate (30 mg, 96.8%). LC-MS (ESI) [M+H] + 615.3.
步驟4. 根據通用步驟E描述的方法，合成為黃色固體的標題產物（70 ）（10.3mg，61.0%）。LC-MS (ESI) [M+H]+ 415.2.1 H NMR (400 MHz, DMSO-d6 ): δ 11.48 (s, 1H), 8.52 (s, 1H), 7.96 (s, 1H), 7.67 (s, 1H), 7.58 (s, 1H), 7.31 (s, 1H), 6.87 (d, J = 1.6 Hz, 1H), 6.69 - 6.64 (m, 3H), 5.23 (s, 2H), 3.78 (s, 3H), 2.50 - 2.23 (m, 2H), 2.01 (s, 3H), 1.44 - 1.41 (m, 1H), 1.25 - 1.21 (m, 1H)。Step 4. According to the method described in general step E, the title product ( 70 ) (10.3 mg, 61.0%) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 415.2. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.48 (s, 1H), 8.52 (s, 1H), 7.96 (s, 1H), 7.67 (s, 1H), 7.58 (s, 1H), 7.31 (s, 1H), 6.87 (d, J = 1.6 Hz, 1H), 6.69-6.64 (m, 3H), 5.23 (s, 2H), 3.78 ( s, 3H), 2.50-2.23 (m, 2H), 2.01 (s, 3H), 1.44-1.41 (m, 1H), 1.25-1.21 (m, 1H).
N-(8- 氨基 -7- 氰 -6-(4- 甲基吡啶 -3- 基 ) 噌啉 -3- 基 ) 環丙烷甲醯胺（ 71 ）的製備 
步驟 1 .在0℃條件下，將NIS（N-碘代琥珀醯亞胺）（7.00mg，0.0312mmol）慢慢地加入到N-(8-氨基-6-(4-甲基吡啶-3-基)噌啉-3-基)環丙烷甲醯胺(1 )（10.0mg，0.0312mmol）的DMF（2mL）溶液中。將反應混合物在25℃條件下攪拌3小時。將反應混合物濃縮。利用prep-TLC（DCM/MeOH=20/1）純化殘餘物，得到為黃色固體的產物（6.00mg，43.2%）。LC-MS (ESI) [M+H]+ 446.2.1 H NMR (400 MHz, DMSO-d6 ): δ 11.72 (s, 1H), 8.83 (s, 1H), 8.51 (d, J = 5.2 Hz, 1H), 8.28 (s, 1H), 7.40 (d, J = 5.2 Hz, 1H), 6.97 (s, 2H), 6.70 (s, 1H), 2.20 – 2.18 (m, 1 H), 2.07 (s, 3H), 0.94 – 0.89 (m, 4 H)。 Step 1. At 0℃, slowly add NIS (N-iodosuccinimide) (7.00mg, 0.0312mmol) to N-(8-amino-6-(4-methylpyridine-3) -Cinolin-3-yl)cyclopropanecarboxamide ( 1 ) (10.0mg, 0.0312mmol) in DMF (2mL). The reaction mixture was stirred at 25°C for 3 hours. The reaction mixture was concentrated. The residue was purified by prep-TLC (DCM/MeOH=20/1) to obtain the product (6.00 mg, 43.2%) as a yellow solid. LC-MS (ESI) [M+H] + 446.2. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.72 (s, 1H), 8.83 (s, 1H), 8.51 (d, J = 5.2 Hz , 1H), 8.28 (s, 1H), 7.40 (d, J = 5.2 Hz, 1H), 6.97 (s, 2H), 6.70 (s, 1H), 2.20 – 2.18 (m, 1 H), 2.07 (s , 3H), 0.94 – 0.89 (m, 4 H).
步驟 2 . 在Ar條件下，將在DMF（0.5mL）中的N-(8-氨基-7-碘-6-(4-甲基吡啶-3-基)噌啉-3-基)環丙烷甲醯胺（5.00mg，0.0112mmol）、Pd(t-Bu3 P)2 （1.00 mg, 0.00170 mmol）、 Zn(CN)2 （3.00 mg, 0.0224 mmol）和 Zn （0.30 mg，0.00390 mmol）的混合物在110℃條件下攪拌16小時。將混合物過濾。利用prep-HPLC對濾液進行純化，得到為黃色固體的產物（3.70mg，96%產率）。LC-MS (ESI) [M+H]+ 345.1.1 H NMR (400 MHz, MeOD-d4 ): δ 8.95 (s, 1H), 8.86 – 8.85 (m, 2H), 8.16 (d, J = 6.0 Hz, 1H), 6.77 (s, 1H), 2.61 (s, 3H), 2.06 – 2.04 (m, 1H), 1.08 – 0.97 (m, 4 H)。 Step 2. Under Ar conditions, add N-(8-amino-7-iodo-6-(4-methylpyridin-3-yl)cinolin-3-yl)cyclopropane in DMF (0.5mL) Formamide (5.00mg, 0.0112mmol), Pd(t-Bu 3 P) 2 (1.00 mg, 0.00170 mmol), Zn(CN) 2 (3.00 mg, 0.0224 mmol) and Zn (0.30 mg, 0.00390 mmol) The mixture was stirred at 110°C for 16 hours. The mixture was filtered. The filtrate was purified by prep-HPLC to obtain the product as a yellow solid (3.70 mg, 96% yield). LC-MS (ESI) [M+H] + 345.1. 1 H NMR (400 MHz, MeOD-d 4 ): δ 8.95 (s, 1H), 8.86 – 8.85 (m, 2H), 8.16 (d, J = 6.0 Hz, 1H), 6.77 (s, 1H), 2.61 (s, 3H), 2.06 – 2.04 (m, 1H), 1.08 – 0.97 (m, 4 H).
四氫 -2H- 吡喃 -4- 基 (8- 氨基 -6-(5- 氨基 -4- 甲基吡啶 -3- 基 ) 噌啉 )-3- 基 ) 氨基甲酸酯（ 72 ）的製備 
步驟1. 在0℃條件下，將氯甲酸-4-硝基苯酯（1.97g，9.80mol）加入到四氫-2H-吡喃-4-醇（1.00g，9.80mol）和TEA（1.19g，11.7mmol）的DCM（40.0mL）溶液中，生成的產物在室溫下攪拌2h。將混合物用水（20mL）稀釋，並用EtOAc (40 mL x 3)萃取。合併的有機層用Na2 SO4 乾燥，並在真空條件下濃縮得到殘餘物，並通過矽膠柱色譜法（PE:EA=10:1 到 5:1，v/v）純化，得到為白色固體的標題產物（2.40g，產率91.7%）。1 H NMR (400 MHz, CDCl3 ): δ 8.30 – 8.28 (m, 1H), 7.41 – 7.26 (m, 1H), 4.98 – 4.92 (m, 1H), 4.00 – 3.97 (m, 2H), 3.61 – 3.55 (m, 2H), 2.06 – 2.05 (m, 2H), 1.87 – 1.83 (m, 2H)。Step 1. Add 4-nitrophenyl chloroformate (1.97g, 9.80mol) to tetrahydro-2H-pyran-4-ol (1.00g, 9.80mol) and TEA (1.19 g, 11.7mmol) in DCM (40.0mL) solution, the resulting product was stirred at room temperature for 2h. The mixture was diluted with water (20 mL) and extracted with EtOAc (40 mL x 3). The combined organic layer was dried with Na 2 SO 4 and concentrated under vacuum to obtain a residue, which was purified by silica gel column chromatography (PE:EA=10:1 to 5:1, v/v) to obtain a white solid The title product (2.40g, 91.7% yield). 1 H NMR (400 MHz, CDCl 3 ): δ 8.30 – 8.28 (m, 1H), 7.41 – 7.26 (m, 1H), 4.98 – 4.92 (m, 1H), 4.00 – 3.97 (m, 2H), 3.61 – 3.55 (m, 2H), 2.06 – 2.05 (m, 2H), 1.87 – 1.83 (m, 2H).
步驟2. 在室溫條件下，將K2 CO3 (455 mg, 3.29 mol)和(叔丁氧基羰基)(5-(8-氯-3-氨基噌啉-6-基)-4-甲基吡啶-3-基)氨基甲酸叔丁基酯（439mg，1.64mol）加入到(四氫-2H-吡喃-4-基)4-硝基苯基碳酸酯（160mg，0.329mol）的DMF溶液中。生成的產物在70℃條件下攪拌16小時。冷卻至室溫後，將混合物用水（20mL）稀釋，並用EA（20 mL X 3）萃取。合併的有機層用鹽水（15 mL x 3）洗滌，用Na2 SO4 乾燥，過濾，並在真空下濃縮得到殘餘物，用prep-TLC（PE:EA = 1:2, v/v）純化得到為黃色固體的標題產物（60.0 mg, 29.7%）。LC-MS (ESI) [M+H]+ 614.2。Step 2. At room temperature, combine K 2 CO 3 (455 mg, 3.29 mol) and (tert-butoxycarbonyl)(5-(8-chloro-3-aminocinnoline-6-yl)-4- (Methylpyridin-3-yl) tert-butyl carbamate (439mg, 1.64mol) was added to (tetrahydro-2H-pyran-4-yl)4-nitrophenyl carbonate (160mg, 0.329mol) DMF solution. The resulting product was stirred at 70°C for 16 hours. After cooling to room temperature, the mixture was diluted with water (20 mL) and extracted with EA (20 mL X 3). The combined organic layer was washed with brine (15 mL x 3), dried over Na 2 SO 4 , filtered, and concentrated under vacuum to obtain a residue, which was purified by prep-TLC (PE:EA = 1:2, v/v) The title product (60.0 mg, 29.7%) was obtained as a yellow solid. LC-MS (ESI) [M+H] + 614.2.
步驟3. 根據通用步驟A 描述的方法，合成為黃色固體的標題產物（25mg，40.7%）。LC-MS (ESI) [M+H]+ 759.2。Step 3. According to the method described in General Procedure A , the title product (25 mg, 40.7%) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 759.2.
步驟4. 根據通用步驟C描述的方法，合成為黃色固體的所希望的化合物（30mg，粗產物），將其用於下一步驟，無需進一步純化。Step 4. According to the method described in General Step C, the desired compound (30 mg, crude product) was synthesized as a yellow solid and used in the next step without further purification.
步驟5. 根據通用步驟E 描述的方法，合成為白色固體的標題產物（72）（1.44mg，7.72%）。LC-MS (ESI) [M+H]+ 395.2.1 H NMR (400 MHz, MeOD-d4 ): δ 8.37 (s, 1H), 7.96 (s, 1H), 7.74 (s, 1H), 6.91 (s, 3H ), 6.75 (s, 1H ), 5.03 – 5.01 (m, 1H), 3.99 – 3.96 (m, 2H), 3.63 – 3.58 (m ,2H), 2.13 (s, 3H), 2.07 – 2.03 (m, 2H), 1.80 – 1.78 (m, 2H)。Step 5. According to the method described in General Procedure E , the title product (72) (1.44 mg, 7.72%) was synthesized as a white solid. LC-MS (ESI) [M+H] + 395.2. 1 H NMR (400 MHz, MeOD-d 4 ): δ 8.37 (s, 1H), 7.96 (s, 1H), 7.74 (s, 1H), 6.91 (s, 3H ), 6.75 (s, 1H ), 5.03 – 5.01 (m, 1H), 3.99 – 3.96 (m, 2H), 3.63 – 3.58 (m ,2H), 2.13 (s, 3H), 2.07 – 2.03 (m, 2H), 1.80 – 1.78 (m, 2H).
(1R,2R)-N-(8- 氨基 -7- 氟 -6-(8- 甲基 -2,3- 二氫 -1H- 吡啶並 [2,3-b][1,4] 惡嗪 -7- 基 ) 噌啉 -3- 基 )-2-(1- 甲基 -1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺（ 73a ）和 (1S,2S)-N-(8- 氨基 -7- 氟 -6-(8- 甲基 -2,3- 二氫 -1H- 吡啶並 [2,3-b][1,4] 惡嗪 -7- 基 ) 噌啉 -3- 基 )-2-(1- 甲基 -1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺（ 73b ）的製備 
步驟1. 根據通用步驟D 描述的方法（90℃，2h，利用(1-(叔丁氧基羰基)-8-甲基-2,3-二氫-1H-吡啶並[2,3-b][1,4]惡嗪-7-基)硼酸），合成為黃色固體的7-(8-氯-7-氟-3-硝基噌啉-6-基)-8-甲基-2,3-二氫-1H-吡啶並[2,3-b][1,4]惡嗪-1-羧酸叔丁基酯（400mg，64.0%）。LC-MS (ESI) [M+Na]+ 498.1。Step 1. According to the method described in general step D (90℃, 2h, using (1-(tert-butoxycarbonyl)-8-methyl-2,3-dihydro-1H-pyrido[2,3-b) ][1,4]oxazine-7-yl)boronic acid), 7-(8-chloro-7-fluoro-3-nitrocinnoline-6-yl)-8-methyl-2 synthesized as a yellow solid ,3-Dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl ester (400mg, 64.0%). LC-MS (ESI) [M+Na] + 498.1.
步驟2. 在Ar條件下，將在EtOH/H2 O (20/2 mL)溶劑體系中的7-(8-氯-7-氟-3-硝基噌啉-6-基)-8-甲基-2,3-二氫-1H-吡啶並[2,3-b][1,4]惡嗪-1-羧酸叔丁基酯（580mg，1.22mmol）、Fe（546 mg, 9.75 mmol）和NH4 Cl (522 mg, 9.75 mmol)在90℃條件下加熱16小時。將混合物冷卻至室溫。將混合物用NaHCO3 水溶液（30mL）稀釋，用乙酸乙酯（30 mL X 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓條件下濃縮。粗產物（500mg，91.9%）直接用於下一步驟。LC-MS (ESI) [M+H]+ 448.1。Step 2. Under Ar conditions, add 7-(8-chloro-7-fluoro-3-nitrocinnoline-6-yl)-8- in EtOH/H 2 O (20/2 mL) solvent system Methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl ester (580mg, 1.22mmol), Fe (546 mg, 9.75 mmol) and NH 4 Cl (522 mg, 9.75 mmol) were heated at 90°C for 16 hours. The mixture was cooled to room temperature. The mixture was diluted with aqueous NaHCO 3 (30 mL) and extracted with ethyl acetate (30 mL×3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product (500 mg, 91.9%) was used directly in the next step. LC-MS (ESI) [M+H] + 448.1.
步驟3. 將在甲苯（8mL）中的7-(3-氨基-8-氯-7-氟-1,2-二氫噌啉-6-基)-8-甲基-2,3-二氫-1H-吡啶並[2,3-b][1,4]惡嗪-1-羧酸叔丁基酯（460mg，1.03mmol）和Pb(OAc)4 (455 mg， 1.03 mmol)的混合物在120℃條件下加熱1小時。將混合物在減壓條件下濃縮。利用矽膠柱色譜法（DCM/MeOH=50/1）純化，得到為黃色固體的產物（260 mg, 57.0%）。LC-MS (ESI) [M+H]+ 446.3.1 H NMR (400 MHz, DMSO-d6 ): δ 7.98 (s, 1H), 7.74 (d, J = 8.0 Hz, 1H), 7.09 (s, 1H), 6.84 (s, 2H), 4.68 – 4.18 (m, 3H), 3.20 (br s, 1H), 1.91 (s, 3H), 1.47 (s, 9H)。Step 3. Add 7-(3-amino-8-chloro-7-fluoro-1,2-dihydrocinoline-6-yl)-8-methyl-2,3-di in toluene (8mL) Mixture of hydrogen-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl ester (460 mg, 1.03 mmol) and Pb(OAc) 4 (455 mg, 1.03 mmol) Heat at 120°C for 1 hour. The mixture was concentrated under reduced pressure. Purified by silica gel column chromatography (DCM/MeOH=50/1), the product (260 mg, 57.0%) was obtained as a yellow solid. LC-MS (ESI) [M+H] + 446.3. 1 H NMR (400 MHz, DMSO-d 6 ): δ 7.98 (s, 1H), 7.74 (d, J = 8.0 Hz, 1H), 7.09 (s , 1H), 6.84 (s, 2H), 4.68 – 4.18 (m, 3H), 3.20 (br s, 1H), 1.91 (s, 3H), 1.47 (s, 9H).
步驟4. 根據與製備69 描述的相似的步驟，合成希望的產物（260mg，75.0%，粗物），將其直接用於下一步驟。LC-MS (ESI) [M+H]+ 594.4。Step 4. Following a similar procedure as described in Preparation 69 , synthesize the desired product (260 mg, 75.0%, crude) and use it directly in the next step. LC-MS (ESI) [M+H] + 594.4.
步驟5. 根據通用步驟A 描述的方法，合成為黃色固體的所希望的產物（40.0mg，14.6%）。LC-MS (ESI) [M+H]+ 739.2。Step 5. According to the method described in General Step A , the desired product (40.0 mg, 14.6%) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 739.2.
步驟6. 根據通用步驟C 描述的方法，合成為黃色固體的所希望的產物（25 mg，80.4%）。LC-MS (ESI) [M+H]+ 575.1。Step 6. According to the method described in general step C , the desired product (25 mg, 80.4%) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 575.1.
步驟7. 根據通用步驟E描述的方法，合成為黃色固體的所希望的產物（10.0 mg, 48.4%）。LC-MS (ESI) [M+H]+ 475.1。Step 7. According to the method described in general step E, the desired product (10.0 mg, 48.4%) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 475.1.
手性拆分。 利用SFC（System：Waters SFC 80；柱名稱：Daicel chiralpak IB；柱規格：250*25mm 10 μm；流動相A：Supercritical CO2 ；流動相 B：MeOH (0.1% NH3 ∙H2 O)；A:B: 45:55；波長：214nm；流動速率：70 g/min；柱溫： 25ºC (RT); 背壓: 100bar; 注射：3.6mL；迴圈時間：27 min；樣品溶液製備：將樣品溶解在大約 13ml MeOH中）分離(反式)-N-(8-氨基-7-氟-6(8-甲基-2,3-二氫-1H-吡啶並[2,3-b][1,4]惡嗪-7-基)噌啉-3-基)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（8.00mg，0.0169mmol）固體，得到為黃色固體的(1R,2R)-N-(8-氨基-7-氟-6-(8-甲基-2,3-二氫-1H-吡啶並[2,3-b][1,4]惡嗪-7-基)噌啉-3-基)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（1.03mg，73a ）和為黃色固體的(1S,2S)-N-(8-氨基-7-氟-6-(8-甲基-2,3-二氫-1H-吡啶並[2,3-b][1,4]惡嗪-7-基)噌啉-3-基)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（1.38mg，73b ）。 Chiral split. Use SFC (System: Waters SFC 80; column name: Daicel chiralpak IB; column size: 250*25mm 10 μm; mobile phase A: Supercritical CO 2 ; mobile phase B: MeOH (0.1% NH 3 ∙H 2 O); A :B: 45:55; Wavelength: 214nm; Flow rate: 70 g/min; Column temperature: 25ºC (RT); Back pressure: 100bar; Injection: 3.6mL; Loop time: 27 min; Sample solution preparation: Sample Dissolve in about 13ml MeOH) separate (trans)-N-(8-amino-7-fluoro-6(8-methyl-2,3-dihydro-1H-pyrido[2,3-b][ 1,4]oxazin-7-yl)cinnoline-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-carboxamide (8.00mg, 0.0169mmol ) As a solid to obtain (1R, 2R)-N-(8-amino-7-fluoro-6-(8-methyl-2,3-dihydro-1H-pyrido[2,3-b) as a yellow solid ][1,4]oxazine-7-yl)cinolin-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-carboxamide (1.03mg, 73a ) and (1S,2S)-N-(8-amino-7-fluoro-6-(8-methyl-2,3-dihydro-1H-pyrido[2,3-b] as a yellow solid) [1,4]oxazin-7-yl)cinolin-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-carboxamide (1.38mg, 73b ).
(1R,2R)-N-(8-氨基-7-氟-6-(8-甲基-2,3-二氫-1H-吡啶並[2,3-b][1,4]惡嗪-7-基)噌啉-3-基)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（73a ）。LC-MS (ESI) [M+H]+ 475.5。SFC方法注釋: CO2 /MeOH/DEA 60/40/0.04 1.8ml/min IB, 3um, 3*100 (Daicel). RT = 2.763 min, e.e% =100%.1 H NMR (400 MHz, DMSO-d6 ): δ 11.55 (s, 1H), 8.60 (s, 1H), 7.59 (s, 1H), 7.33 (d, J=12.8Hz, 2H), 6.96 (d, J=6.8Hz, 1H), 6.52 (s, 2H), 5.73 (s, 1H), 4.30 (s, 2H), 3.78 (s, 3H), 3.35 – 3.34 (m, 2H), 2.28 – 2.25 (s, 2H),1.93 (s, 3H), 1.44 – 1.41 (m, 1H) ,1.26 – 1.23 (m, 1H)。(1R,2R)-N-(8-amino-7-fluoro-6-(8-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine -7-yl)cinnolin-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-carboxamide ( 73a ). LC-MS (ESI) [M+H] + 475.5. SFC method note: CO 2 /MeOH/DEA 60/40/0.04 1.8ml/min IB, 3um, 3*100 (Daicel). RT = 2.763 min, ee% =100%. 1 H NMR (400 MHz, DMSO- d 6 ): δ 11.55 (s, 1H), 8.60 (s, 1H), 7.59 (s, 1H), 7.33 (d, J=12.8Hz, 2H), 6.96 (d, J=6.8Hz, 1H), 6.52 (s, 2H), 5.73 (s, 1H), 4.30 (s, 2H), 3.78 (s, 3H), 3.35 – 3.34 (m, 2H), 2.28 – 2.25 (s, 2H), 1.93 (s, 3H), 1.44 – 1.41 (m, 1H), 1.26 – 1.23 (m, 1H).
(1S,2S)-N-(8-氨基-7-氟-6-(8-甲基-2,3-二氫-1H-吡啶並[2,3-b][1,4]惡嗪-7-基)噌啉-3-基)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（73b ）。LC-MS (ESI) [M+H]+ 475.5. SFC方法注釋: CO2 /MeOH/DEA 60/40/0.04 1.8ml/min IB, 3um, 3*100 (Daicel). RT = 3.726 min, e.e% =100%.1 H NMR (400 MHz, DMSO-d6 ): δ 11.55 (s, 1H), 8.60 (s, 1H), 7.59 (s, 1H), 7.33 (d, J=12.8Hz, 2H), 6.96 (d, J=6.0Hz, 1H), 6.53 (s, 2H), 5.73 (s, 1H), 4.30 (s, 2H), 3.78 (s, 3H), 3.35 – 3.34 (m, 2H), 2.27 (s, 2H),1.93 (s, 3H), 1.43 (s, 1H), 1.23 (s, 1H)。(1S,2S)-N-(8-amino-7-fluoro-6-(8-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine -7-yl)cinnolin-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-carboxamide ( 73b ). LC-MS (ESI) [M+H] + 475.5. SFC method note: CO 2 /MeOH/DEA 60/40/0.04 1.8ml/min IB, 3um, 3*100 (Daicel). RT = 3.726 min, ee % =100%. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.55 (s, 1H), 8.60 (s, 1H), 7.59 (s, 1H), 7.33 (d, J=12.8Hz, 2H ), 6.96 (d, J=6.0Hz, 1H), 6.53 (s, 2H), 5.73 (s, 1H), 4.30 (s, 2H), 3.78 (s, 3H), 3.35 – 3.34 (m, 2H) , 2.27 (s, 2H), 1.93 (s, 3H), 1.43 (s, 1H), 1.23 (s, 1H).
(( 反式Trans )-N-(8-)-N-(8- 氨基Amino -6-(3,8--6-(3,8- 二甲基Dimethyl -2,3--2,3- 二氫Dihydro -1H--1H- 吡啶並Pyrido [2,3-b][1,4][2,3-b][1,4] 惡嗪Oxazine -7--7- 基base )-7-)-7- 氟噌啉Flucinoline -3--3- 基base )-2-(1-)-2-(1- 甲基methyl -1H--1H- 吡唑Pyrazole -4--4- 基base )) 環丙烷Cyclopropane -1--1- 甲醯胺（Formamide ( 7474 ）的製備) Preparation
根據與化合物 73a 和73b 相似的方法，利用(1-(叔丁氧基羰基)-3,8-二甲基-2,3-二氫-1H-吡啶並[2,3-b][1,4]惡嗪-7-基)硼酸製備為黃色固體的化合物74（14.4 mg, 57.8%）。LC-MS (ESI) [M+H]+ 489.6.1 H NMR (400 MHz, DMSO-d6 ): δ 11.54 (s, 1H), 8.59 (s, 1H), 7.58 (s, 1H), 7.35 (s, 1H), 7.31 (s, 1H), 6.96 (d, J = 6.8 Hz, 1H), 6.51 (s, 2H), 5.73 (s, 1H), 4.30 – 4.27 (m, 1H), 3.77 (s, 3H), 3.49 – 3.42 (m, 1H), 3.02 – 2.99 (m, 1H), 2.33 – 2.23 (m, 2H), 1.94 (s, 3H), 1.44 – 1.43 (m, 1H), 1.42 (d, J = 4.8 Hz, 3H), 1.24 – 1.22 (m, 1H)。According to a method similar to compound 73a and 73b , using (1-(tert-butoxycarbonyl)-3,8-dimethyl-2,3-dihydro-1H-pyrido[2,3-b][1 ,4]oxazine-7-yl)boronic acid was prepared as yellow solid compound 74 (14.4 mg, 57.8%). LC-MS (ESI) [M+H] + 489.6. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.54 (s, 1H), 8.59 (s, 1H), 7.58 (s, 1H), 7.35 (s, 1H), 7.31 (s, 1H), 6.96 (d, J = 6.8 Hz, 1H), 6.51 (s, 2H), 5.73 (s, 1H), 4.30 – 4.27 (m, 1H), 3.77 ( s, 3H), 3.49 – 3.42 (m, 1H), 3.02 – 2.99 (m, 1H), 2.33 – 2.23 (m, 2H), 1.94 (s, 3H), 1.44 – 1.43 (m, 1H), 1.42 ( d, J = 4.8 Hz, 3H), 1.24 – 1.22 (m, 1H).
( 反式 )-N-(8- 氨基 -6-(1,5- 二甲基 -1H- 吡唑 -4- 基 )-7- 氟噌啉 -3- 基 )-2-(1- 甲基 -1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺（ 75 ）的製備 
步驟1. 根據通用步驟D 描述的方法，合成為黃色固體的8-氯-6-(1,5-二甲基-1H-吡唑-4-基)-7-氟噌啉-3胺（215 mg, 67.2%）。LC-MS (ESI) [M+Na]+ 292.1。Step 1. According to the method described in general step D , 8-chloro-6-(1,5-dimethyl-1H-pyrazol-4-yl)-7-fluorocinoline-3amine ( 215 mg, 67.2%). LC-MS (ESI) [M+Na] + 292.1.
根據製備化合物 73a/73b 相似的方法製備為黃色固體的化合物 75 （16.7 mg, 46.4%）。LC-MS (ESI) [M+H]+ 421.5.1 H NMR (400 MHz, DMSO-d6 ): δ 11.51 (s, 1H), 8.59 (s, 1H), 7.62 (d, J = 2.0 Hz, 1H), 7.59 (s, 1H), 7.32 (s, 1H), 7.00 (d, J = 7.2 Hz, 1H), 6.46 (s, 2H), 3.83 (s, 3H), 3.78 (s, 3H), 2.37 (s, 3H), 2.29 – 2.22 (m, 2H), 1.43 – 1.41 (m, 1H), 1.24 – 1.23 (m, 1H)。 The compound 75 (16.7 mg, 46.4%) was prepared as a yellow solid according to a method similar to the preparation of compound 73a/73b. LC-MS (ESI) [M+H] + 421.5. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.51 (s, 1H), 8.59 (s, 1H), 7.62 (d, J = 2.0 Hz , 1H), 7.59 (s, 1H), 7.32 (s, 1H), 7.00 (d, J = 7.2 Hz, 1H), 6.46 (s, 2H), 3.83 (s, 3H), 3.78 (s, 3H) , 2.37 (s, 3H), 2.29 – 2.22 (m, 2H), 1.43 – 1.41 (m, 1H), 1.24 – 1.23 (m, 1H).
( 反式 )-N-(8- 氨基 -6-(8- 甲基 -2- 氧代 -2,3- 二氫 -1H- 吡啶並 [2,3-b][1,4] 惡嗪 -7- 基 ) 噌啉 -3- 基 )-2-(1- 甲基 -1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺（ 76 ）的製備 
步驟1&2. 在0℃條件下，將NaNO2 （1.62g，23.5mmol）的4mL H2 O溶液慢慢地加入到2-溴-4-氯苯胺（4.85g，23.5mmol）的4.0M HCl（23.5mL）溶液中。將混合物在0℃條件下攪拌1小時。然後在0℃條件下，加入(E)-N,N-二甲基-2-硝基乙烷-1-胺（3.00g，25.9mmol）的乙腈（15mL）溶液。將反應混合物加熱至室溫，並攪拌16h。用水（100mL）稀釋反應混合物。對生成的懸浮液進行過濾。用冷的Et2 O (50 mL)對濾餅進行洗滌，並乾燥得到為橙色固體的(Z)-2-((Z)-(2-溴-4-氯苯基)重氮烯基)-2-硝基乙烷-1-醇（5.40g，17.7mmol，75.0%）。1 H NMR (400 MHz, DMSO-d6 ): δ 13.7 (br s, 1H), 11.1 (s, 1H), 7.97 (d, J = 2.0 Hz, 1H), 7.75 (d, J = 8.8 Hz, 1H), 7.66 - 7.63 (m, 2H)。Steps 1&2. At 0℃, add NaNO 2 (1.62g, 23.5mmol) in 4mL H 2 O solution slowly to 2-bromo-4-chloroaniline (4.85g, 23.5mmol) in 4.0M HCl( 23.5mL) solution. The mixture was stirred at 0°C for 1 hour. Then, at 0°C, a solution of (E)-N,N-dimethyl-2-nitroethane-1-amine (3.00 g, 25.9 mmol) in acetonitrile (15 mL) was added. The reaction mixture was heated to room temperature and stirred for 16 h. The reaction mixture was diluted with water (100 mL). The resulting suspension is filtered. The filter cake was washed with cold Et 2 O (50 mL) and dried to obtain (Z)-2-((Z)-(2-bromo-4-chlorophenyl)diazenyl) as an orange solid -2-nitroethane-1-ol (5.40g, 17.7mmol, 75.0%). 1 H NMR (400 MHz, DMSO-d 6 ): δ 13.7 (br s, 1H), 11.1 (s, 1H), 7.97 (d, J = 2.0 Hz, 1H), 7.75 (d, J = 8.8 Hz, 1H), 7.66-7.63 (m, 2H).
步驟3. 將(Z)-2-((Z)-(2-溴-4-氯苯基)重氮烯基)-2-硝基乙烷-1-醇（178g，582mmol）的PPA（聚磷酸）（900mL）溶液在160℃條件下攪拌5h。將反應混合物倒在冰上，然後用乙酸乙酯（500 mL X 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓條件下濃縮。利用矽膠柱色譜法（石油醚/乙酸乙酯=2/1）純化，得到為黃色固體的8-溴-6-氯-3-硝基噌啉（30.0g，104mmol，18.0%）。Step 3. Add (Z)-2-((Z)-(2-bromo-4-chlorophenyl)diazenyl)-2-nitroethane-1-ol (178g, 582mmol) in PPA ( The solution of polyphosphoric acid (900mL) was stirred at 160°C for 5h. The reaction mixture was poured on ice, and then extracted with ethyl acetate (500 mL X 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Purified by silica gel column chromatography (petroleum ether/ethyl acetate=2/1), 8-bromo-6-chloro-3-nitrocinoline (30.0 g, 104 mmol, 18.0%) was obtained as a yellow solid.
步驟4.根據通用步驟A 描述的方法，合成為黃色固體的標題產物（69.0mg，0.178mmol，27.0%）。1 H NMR (400 MHz, DMSO-d6 ): δ 9.16 (s, 1H), 8.07 (d, J = 2.0 Hz, 1H), 7.61 – 7.24 (m, 11H)。Step 4. According to the method described in General Procedure A , the title product (69.0 mg, 0.178 mmol, 27.0%) was synthesized as a yellow solid. 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.16 (s, 1H), 8.07 (d, J = 2.0 Hz, 1H), 7.61 – 7.24 (m, 11H).
步驟5. 根據通用步驟C 描述的方法，合成為黃色油狀物的標題產物（314mg，1.40mmol，81.8%）。LC-MS (ESI) [M+H]+ 225.1。Step 5. According to the method described in General Procedure C , the title product (314 mg, 1.40 mmol, 81.8%) was synthesized as a yellow oil. LC-MS (ESI) [M+H] + 225.1.
步驟6. 在室溫條件下，將Boc2 O (413 mg, 1.90 mmol)和 DMAP (3.10 mg, 0.0253 mmol)加入到6-氯-3-硝基噌啉-8-胺（283mg，1.26mmol）和TEA（255mg，2.53mmol）的DCM（10mL）溶液中。將混合物在室溫下攪拌2h。將混合物濃縮，並利用矽膠柱色譜法（PE: EA=100:1-30:1）純化得到為棕色油狀物的(叔丁氧基羰基)(6-氯-3-硝基噌啉-8-基)氨基甲酸叔丁基酯。LC-MS (ESI) [M-Boc+H]+ 325.0。Step 6. At room temperature, add Boc 2 O (413 mg, 1.90 mmol) and DMAP (3.10 mg, 0.0253 mmol) to 6-chloro-3-nitrocinnoline-8-amine (283 mg, 1.26 mmol) ) And TEA (255mg, 2.53mmol) in DCM (10mL) solution. The mixture was stirred at room temperature for 2h. The mixture was concentrated and purified by silica gel column chromatography (PE: EA=100:1-30:1) to obtain (tert-butoxycarbonyl)(6-chloro-3-nitrocinoline- 8-yl) tert-butyl carbamate. LC-MS (ESI) [M-Boc+H] + 325.0.
步驟7. 在H2 條件下，將(叔丁氧基羰基)(6-氯-3-硝基噌啉-8-基)氨基甲酸叔丁基酯（90.0 mg, 0.212 mmol）和Pt2 O (8.00 mg) 的EA（50mL）溶液在室溫條件下攪拌3h。將混合物過濾，濾液進行濃縮，並利用prep-TLC（PE:EA=1:1）純化得到為棕色油狀物的 (3-氨基-6-氯噌啉-8-基) (叔丁氧基羰基)氨基甲酸叔丁基酯（66mg，0.168mmo，79.2%）。LC-MS (ESI) [M+H]+ 395.0。Step 7. Under H 2 conditions, combine tert-butyl (tert-butoxycarbonyl)(6-chloro-3-nitrocinnoline-8-yl)carbamate (90.0 mg, 0.212 mmol) and Pt 2 O (8.00 mg) of EA (50mL) solution was stirred at room temperature for 3h. The mixture was filtered, the filtrate was concentrated, and purified by prep-TLC (PE:EA=1:1) to obtain (3-amino-6-chlorocinoline-8-yl) (tert-butoxy (Carbonyl) tert-butyl carbamate (66 mg, 0.168 mmo, 79.2%). LC-MS (ESI) [M+H] + 395.0.
步驟8. 在Ar和0℃條件下，將POCl3 (40.0 mg, 0.260 mmol) 加入到(3-氨基-6-氯噌啉-8-基)(叔丁氧基羰基)氨基甲酸叔丁基酯（31.0mg，0.0787mmol）、反式-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-羧酸（26.0mg，0.157mmol）和TEA（160mg，1.57mmol）的DCM（5mL）溶液中。將混合物在室溫下攪拌16h。然後用水（50mL）稀釋，用EA（30 mL x 3）萃取。有機層用鹽水（50 mL x 3）洗滌，用無水Na2 SO4 乾燥，過濾，濃縮並用prep-TLC（DCM:MeOH=20:1）純化得到為黃色固體的(叔丁氧基羰基)(6-氯-3-(反式-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯氨基)噌啉-8-基)氨基甲酸叔丁基酯（12.0 mg, 0.0221 mmol, 28.1 %）。LC-MS (ESI) [M+H]+ 543.3。 Step 8. Add POCl 3 (40.0 mg, 0.260 mmol) to tert-butyl (3-amino-6-chlorocinoline-8-yl)(tert-butoxycarbonyl)carbamate under Ar and 0°C conditions Ester (31.0mg, 0.0787mmol), trans-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-carboxylic acid (26.0mg, 0.157mmol) and TEA (160mg, 1.57mmol) ) In DCM (5mL) solution. The mixture was stirred at room temperature for 16 h. Then it was diluted with water (50 mL) and extracted with EA (30 mL x 3). The organic layer was washed with brine (50 mL x 3), dried over anhydrous Na 2 SO 4 , filtered, concentrated and purified with prep-TLC (DCM:MeOH=20:1) to obtain (tert-butoxycarbonyl) as a yellow solid ( 6-chloro-3-(trans-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-carboxamide)cinoline-8-yl)tert-butyl carbamate ( 12.0 mg, 0.0221 mmol, 28.1 %). LC-MS (ESI) [M+H] + 543.3.
步驟9. 根據通用步驟F描述的方法，合成為黃色油狀物的標題產物（7.00 mg, 0.0104 mmol, 47.3 %）。LC-MS (ESI) [M-Boc+H]+ 571.3。Step 9. According to the method described in General Procedure F, the title product (7.00 mg, 0.0104 mmol, 47.3 %) was synthesized as a yellow oil. LC-MS (ESI) [M-Boc+H] + 571.3.
步驟10. 根據通用步驟E描述的方法，合成為黃色固體的化合物 76 （4.02 mg, 0.00855 mmol, 24.9 %）。LC-MS (ESI) [M+H]+ 471.4.1 H NMR (400 MHz, DMSO-d6 ): δ 11.5 (s, 1H), 8.53 (s, 1H), 7.63 – 7.59 (m, 2H), 7.32 (s, 1H), 6.88 (s, 1H), 6.69 – 6.67 (m, 3H), 4.67 (s, 2H), 3.78 (s, 3H), 2.33 – 2.22 (m, 2H), 2.18 (s, 3H),1.44 – 1.40 (m, 1H), 1.26 – 1.22 (m, 1H)。Step 10. According to the method described in general step E, compound 76 (4.02 mg, 0.00855 mmol, 24.9 %) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 471.4. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.5 (s, 1H), 8.53 (s, 1H), 7.63 – 7.59 (m, 2H) , 7.32 (s, 1H), 6.88 (s, 1H), 6.69 – 6.67 (m, 3H), 4.67 (s, 2H), 3.78 (s, 3H), 2.33 – 2.22 (m, 2H), 2.18 (s , 3H), 1.44 – 1.40 (m, 1H), 1.26 – 1.22 (m, 1H).
( 反式 )-N-(8- 氨基 -6-(2- 氟 -4- 甲基吡啶 -3- 基 ) 噌啉 -3- 基 )-2-(1- 甲基 -1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺（ 77 ）的製備 
步驟1. 根據製備73a/73b 的步驟2描述的方法，合成為黃色固體的標題產物（4.3g，57.7%）。LC-MS (ESI) [M+H]+ 359.1。Step 1. According to the method described in step 2 of the preparation of 73a/73b , the title product (4.3 g, 57.7%) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 359.1.
步驟2. 根據製備73a/73b 的步驟4描述的方法，合成為類白色固體的反式-N-(6-氯-8-((二苯亞甲基)氨基)噌啉-3-基)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（4.0g，94.3%）。LC-MS (ESI) [M+H]+ 507.2。Step 2. According to the method described in step 4 of the preparation of 73a/73b , synthesize trans-N-(6-chloro-8-((benzylidene)amino)cinolin-3-yl) as a white solid -2-(1-Methyl-1H-pyrazol-4-yl)cyclopropane-1-carboxamide (4.0 g, 94.3%). LC-MS (ESI) [M+H] + 507.2.
步驟3. 根據通用步驟F 描述的方法，合成為黃色油狀物的標題產物（48.0mg，52.3%）。LC-MS (ESI) [M+H]+ 582.1。Step 3. According to the method described in General Procedure F , the title product (48.0 mg, 52.3%) was synthesized as a yellow oil. LC-MS (ESI) [M+H] + 582.1.
步驟4. 根據通用步驟C 描述的方法，合成為黃色固體的化合物 77 （25.9mg，72.1%產率）。LC-MS (ESI) [M+H]+ 418.4。1 H NMR (400 MHz, DMSO-d6 ): δ 11.5 (s, 1H), 8.54 (s, 1H), 8.15 (d, J = 5.2 Hz, 1H), 7.59 (s, 1H), 7.37 (d, J = 5.2 Hz, 1H), 7.32 (s, 1H), 6.95 (d, J = 1.2 Hz, 1H), 6.74 (s, 2H), 6.64 (s, 1H), 3.78 (s, 3H), 2.30 – 2.22 (m, 5H), 1.44 – 1.41 (m, 1H), 1.26 – 1.722 (m, 1H)。Step 4. According to the method described in general step C , compound 77 (25.9 mg, 72.1% yield) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 418.4. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.5 (s, 1H), 8.54 (s, 1H), 8.15 (d, J = 5.2 Hz, 1H), 7.59 (s, 1H), 7.37 (d , J = 5.2 Hz, 1H), 7.32 (s, 1H), 6.95 (d, J = 1.2 Hz, 1H), 6.74 (s, 2H), 6.64 (s, 1H), 3.78 (s, 3H), 2.30 – 2.22 (m, 5H), 1.44 – 1.41 (m, 1H), 1.26 – 1.722 (m, 1H).
(( 反式Trans )-N-(8-)-N-(8- 氨基Amino -6-(5--6-(5- 氰基Cyano -2--2- 甲基苯基Methyl phenyl )) 噌啉Cinolin -3--3- 基base )-2-(1-)-2-(1- 甲基methyl -1H--1H- 吡唑Pyrazole -4--4- 基base )) 環丙烷Cyclopropane -1--1- 甲醯胺（Formamide ( 7878 ）的製備) Preparation
利用與製備化合物 77 相似的方法製備為黃色固體的化合物 78 （21.0mg，36.5%）。LC-MS (ESI) [M+H]+ 424.4。1 H NMR (400 MHz, DMSO-d6 ): δ 11.52 (s, 1H), 8.54 (s, 1H), 7.80 – 7.74 (m, 2H), 7.58 – 7.54 (m, 2H ), 7.31 (s, 1H ), 6.94 (s, 1H), 6.70 (s, 3H), 3.78 (s ,3H), 2.35 (s, 3H), 2.30 – 2.25 (m, 2H), 1.44 – 1.42 (m, 2H), 1.25 – 1.23 (m, 1H)。 The compound 78 (21.0 mg, 36.5%) was prepared as a yellow solid using a method similar to the preparation of compound 77. LC-MS (ESI) [M+H] + 424.4. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.52 (s, 1H), 8.54 (s, 1H), 7.80 – 7.74 (m, 2H), 7.58 – 7.54 (m, 2H ), 7.31 (s, 1H ), 6.94 (s, 1H), 6.70 (s, 3H), 3.78 (s ,3H), 2.35 (s, 3H), 2.30 – 2.25 (m, 2H), 1.44 – 1.42 (m, 2H), 1.25 – 1.23 (m, 1H).
(( 反式Trans )-N-(8-)-N-(8- 氨基Amino -6-(2--6-(2- 氯chlorine -6--6- 氟苯基Fluorophenyl )) 噌啉Cinolin -3--3- 基base )-2-(1-)-2-(1- 甲基methyl -1H--1H- 吡唑Pyrazole -4--4- 基base )) 環丙烷Cyclopropane -1--1- 甲醯胺（Formamide ( 7979 ）的製備) Preparation
利用與製備化合物 77 相似的方法製備為黃色固體的化合物 79 （23.0mg，62.2%）。LC-MS (ESI) [M+H]+ 437.0。1 H NMR (400 MHz, DMSO-d6 ): δ 11.55 (s, 1H), 8.54 (s, 1H), 7.58 (d, J = 2.4 Hz, 1H), 7.53 - 7.49 (m, 2H), 7.40 - 7.37 (m, 1H), 7.32 (s, 1H), 6.95 (s, 1H), 6.74 (s, 2H), 6.66 (s, 1H), 3.78 (s, 3H), 2.29 - 2.23 (m, 2H), 1.44 - 1.42 (m, 1H), 1.27 - 1.15 (m, 1H)。 The compound 79 (23.0 mg, 62.2%) was prepared as a yellow solid using a method similar to the preparation of compound 77. LC-MS (ESI) [M+H] + 437.0. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.55 (s, 1H), 8.54 (s, 1H), 7.58 (d, J = 2.4 Hz, 1H), 7.53-7.49 (m, 2H), 7.40 -7.37 (m, 1H), 7.32 (s, 1H), 6.95 (s, 1H), 6.74 (s, 2H), 6.66 (s, 1H), 3.78 (s, 3H), 2.29-2.23 (m, 2H ), 1.44-1.42 (m, 1H), 1.27-1.15 (m, 1H).
反式Trans -N-(8--N-(8- 氨基Amino -6-(1--6-(1- 甲基methyl -1H--1H- 咪唑Imidazole -5--5- 基base )) 噌啉Cinolin -3--3- 基base )-2-(1-)-2-(1- 甲基methyl -1H--1H- 吡唑Pyrazole -4--4- 基base )) 環丙烷Cyclopropane -1--1- 甲醯胺（Formamide ( 8080 ）的製備) Preparation
利用與製備化合物 77 相似的方法製備為黃色固體的化合物 80 （23.1mg，63.9%）。LC-MS (ESI) [M+H]+ 389.1。1 H NMR (400 MHz, DMSO-d6 ): δ 11.50 (s, 1H), 8.56 (s, 1H), 7.77 (s, 1H), 7.59 (s, 1H), 7.32 (s, 1H), 7.21 (s, 1H), 7.11 (s, 1H), 6.90 (s, 1H), 6.65 (s, 2H), 3.79 (d, J = 8.4 Hz, 6H), 2.44 - 2.24 (m, 2H), 1.44 - 1.41 (m, 1H), 1.24 - 1.22 (m, 1H)。 The compound 80 (23.1 mg, 63.9%) was prepared as a yellow solid by a method similar to the preparation of compound 77. LC-MS (ESI) [M+H] + 389.1. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.50 (s, 1H), 8.56 (s, 1H), 7.77 (s, 1H), 7.59 (s, 1H), 7.32 (s, 1H), 7.21 (s, 1H), 7.11 (s, 1H), 6.90 (s, 1H), 6.65 (s, 2H), 3.79 (d, J = 8.4 Hz, 6H), 2.44-2.24 (m, 2H), 1.44- 1.41 (m, 1H), 1.24-1.22 (m, 1H).
反式Trans -N-(8--N-(8- 氨基Amino -6-(5--6-(5- 氯chlorine -3,4--3,4- 二氫Dihydro -2H--2H- 吡啶並Pyrido [4,3-b] [1,4][4,3-b] [1,4] 惡嗪Oxazine -8--8- 基base )) 噌啉Cinolin -3--3- 基base )-2-(1-)-2-(1- 甲基methyl -1H--1H- 吡唑Pyrazole -4--4- 基base )) 環丙烷Cyclopropane -1--1- 甲醯胺（Formamide ( 8181 ）的製備) Preparation
利用與製備化合物 77 相似的方法製備為黃色固體的化合物 81 （11mg，22.2%）。LC-MS (ESI) [M+H]+ 477.1。1 H NMR (400 MHz, DMSO-d6 ): δ 11.49 (s, 1H), 8.52 (s, 1H), 7.62 (s, 1H), 7.58 (s, 3H ), 7.31 (s, 1H ), 7.05 (s, 1H), 6.90 (s, 1H), 6.61 (s ,2H), 6.02 (s, 1H), 4.29 – 4.23 (m, 2H), 3.46 – 3.40 (m, 2H), 2.29 – 2.48 (m, 2H), 1.40 – 1.45 (m, 1H), 1.25 – 1.22 (m, 1H)。 The compound 81 (11 mg, 22.2%) was prepared as a yellow solid using a method similar to the preparation of compound 77. LC-MS (ESI) [M+H] + 477.1. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.49 (s, 1H), 8.52 (s, 1H), 7.62 (s, 1H), 7.58 (s, 3H ), 7.31 (s, 1H ), 7.05 (s, 1H), 6.90 (s, 1H), 6.61 (s ,2H), 6.02 (s, 1H), 4.29 – 4.23 (m, 2H), 3.46 – 3.40 (m, 2H), 2.29 – 2.48 (m , 2H), 1.40 – 1.45 (m, 1H), 1.25 – 1.22 (m, 1H).
(( 反式Trans )-N-(8-)-N-(8- 氨基Amino -6-(8--6-(8- 甲基methyl -2,3--2,3- 二氫Dihydro -1H--1H- 吡啶並Pyrido [2,3-b][1,4])[2,3-b][1,4]) 噻嗪Thiazide -7--7- 基base )) 噌啉Cinolin -3--3- 基base )-2-(1-)-2-(1- 甲基methyl -1H--1H- 吡唑Pyrazole -4--4- 基base )) 環丙烷Cyclopropane -1--1- 甲醯胺（Formamide ( 8282 ）的製備) Preparation
利用與製備化合物 77 相似的方法製備為黃色固體的化合物 82 （12.0mg，24.4%）。LC-MS (ESI) [M+H]+ 473.1。1 H NMR (400 MHz, DMSO-d6 ): δ 11.49 (s, 1H), 8.52 (s, 1H), 7.58 (s, 1H), 7.57 (s, 1H), 7.31 (s, 1H), 6.85 (d, J = 2.4 Hz, 1H), 6.66 – 6.65 (m, 3H), 5.72 (s, 1H), 3.78 (s ,3H), 3.60 – 3.50 (m, 2H), 3.18 – 3.12 (m, 2H), 2.29 – 2.22 (m, 2H), 1.99 (s, 3H), 1.43 – 1.40 (m, 1H), 1.25 – 1.20 (m, 1H)。 The compound 82 (12.0 mg, 24.4%) was prepared as a yellow solid using a method similar to the preparation of compound 77. LC-MS (ESI) [M+H] + 473.1. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.49 (s, 1H), 8.52 (s, 1H), 7.58 (s, 1H), 7.57 (s, 1H), 7.31 (s, 1H), 6.85 (d, J = 2.4 Hz, 1H), 6.66 – 6.65 (m, 3H), 5.72 (s, 1H), 3.78 (s ,3H), 3.60 – 3.50 (m, 2H), 3.18 – 3.12 (m, 2H) ), 2.29 – 2.22 (m, 2H), 1.99 (s, 3H), 1.43 – 1.40 (m, 1H), 1.25 – 1.20 (m, 1H).
(( 反式Trans )-N-(8-)-N-(8- 氨基Amino -6-(8--6-(8- 甲基methyl -4,4--4,4- 二氧代Dioxo -2,3--2,3- 二氫Dihydro -1H--1H- 吡啶並Pyrido [2,3-b][1,4][2,3-b][1,4] 噻嗪Thiazide -7--7- 基base )) 噌啉Cinolin -3--3- 基base )-2-(1-)-2-(1- 甲基methyl -1H--1H- 吡唑Pyrazole -4--4- 基base )) 環丙烷Cyclopropane -1--1- 甲醯胺Formamide )) （( 8383 ）的製備) Preparation
根據與製備化合物77相似的方法製備為黃色固體的化合物83（7.3mg，19.5%）。LC-MS (ESI) [M+H]+ 505.2。1 H NMR (400 MHz, DMSO-d6 ): δ 11.53 (s, 1H), 8.55 (s, 1H), 7.85 (s, 1H), 7.58 (s, 1H), 7.31 (s, 1H), 6.92 (s, 1H), 6.76 (s, 2H), 6.67 - 6.65 (m, 2H), 3.80 - 3.77 (m, 5H), 3.55 - 3.53 (m, 2H), 2.30 - 2.24 (m, 2H), 2.08 (s, 3H), 1.44 - 1.42 (m, 1H), 1.24 - 1.22 (m, 1H)。The compound 83 (7.3 mg, 19.5%) was prepared as a yellow solid according to a method similar to the preparation of compound 77. LC-MS (ESI) [M+H] + 505.2. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.53 (s, 1H), 8.55 (s, 1H), 7.85 (s, 1H), 7.58 (s, 1H), 7.31 (s, 1H), 6.92 (s, 1H), 6.76 (s, 2H), 6.67-6.65 (m, 2H), 3.80-3.77 (m, 5H), 3.55-3.53 (m, 2H), 2.30-2.24 (m, 2H), 2.08 (s, 3H), 1.44-1.42 (m, 1H), 1.24-1.22 (m, 1H).
( 反式 )-N-(8- 氨基 -6-(5- 甲基 -2- 氧代 -2,3- 二氫惡唑並 [4,5-b] 吡啶 -6- 基 ) 噌啉 -3- 基 )-2-(1- 甲基 -1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺（ 84 ）的製備 
步驟1. 根據標準步驟合成了為黑色固體的(8-氨基-3-反式-2-(1-甲基-1H-吡唑-4-基）環丙烷-1-甲醯氨基)噌啉-6-基)硼酸（80 mg，產率57.6％ ）。LC-MS（ESI）[M + H]+ 353.0。Step 1. According to standard procedures, (8-amino-3-trans-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-methylamino)cinnoline was synthesized as a black solid -6-yl)boronic acid (80 mg, 57.6% yield). LC-MS (ESI) [M + H] + 353.0.
步驟2.根據通用步驟F 中描述的方法（在90℃下保持2小時，使用6-溴-5-甲基惡唑並[4,5-b]吡啶-2-(3H)-酮），合成為黃色固體的標題產物（84 ）（2.19mg，2.43％產率）。LC-MS (ESI) [M+H]+ 457.1。1 H NMR (400 MHz, DMSO-d6 ): δ 11.42 (br s, 1H), 8.48 (s, 1H), 7.58 (s, 1H), 7.31 (s, 1H), 6.86 (s, 2H), 6.76 (d, J = 1.6 Hz, 1H), 6.52 (s, 2H), 6.06 (br s, 1H), 3.78 (s, 3H), 2.32 (s, 3H), 2.30 - 2.22 (m, 2H), 1.45 - 1.40 (m, 1H), 1.24 - 1.20 (m, 1H)。Step 2. According to the method described in General Step F (Keep at 90°C for 2 hours, use 6-bromo-5-methyloxazolo[4,5-b]pyridine-2-(3H)-one), The title product (84 ) (2.19 mg, 2.43% yield) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 457.1. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.42 (br s, 1H), 8.48 (s, 1H), 7.58 (s, 1H), 7.31 (s, 1H), 6.86 (s, 2H), 6.76 (d, J = 1.6 Hz, 1H), 6.52 (s, 2H), 6.06 (br s, 1H), 3.78 (s, 3H), 2.32 (s, 3H), 2.30-2.22 (m, 2H), 1.45-1.40 (m, 1H), 1.24-1.20 (m, 1H).
N-(8- 氨基 -6-(4- 甲基吡啶 -3- 基 ) 噌啉 -3- 基 )-3- 氰基氮雜環丁烷 -1- 甲醯胺（ 85 ）的製備 
步驟1.根據通用步驟F 中描述的方法，合成了為黃色油的8-((二苯基亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-胺（360mg，0.867mmol，69.0％產率）。LC-MS（ESI）[M + H]+ 416.1。Step 1. According to the method described in general step F , 8-((diphenylmethylene)amino)-6-(4-methylpyridin-3-yl)cinoline-3- was synthesized as a yellow oil Amine (360 mg, 0.867 mmol, 69.0% yield). LC-MS (ESI) [M + H] + 416.1.
步驟2. 在0℃下和Ar下，向8-((二苯基亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-胺（200 mg，0.482 mmol）和吡啶（190 mg，2.41 mmol）的DCM（10mL）溶液中加入4-硝基氯甲酸苯酯（117mg，0.578mmol）。將混合物在室溫攪拌2h。該混合物直接用於下一步驟。LC-MS（ESI）[M + H]+ 581.6。Step 2. Add 8-((diphenylmethylene)amino)-6-(4-methylpyridin-3-yl)cinnoline-3-amine (200 mg, 0.482 mmol) and pyridine (190 mg, 2.41 mmol) in DCM (10 mL) was added phenyl 4-nitrochloroformate (117 mg, 0.578 mmol). The mixture was stirred at room temperature for 2h. This mixture was used directly in the next step. LC-MS (ESI) [M + H] + 581.6.
步驟3. 在0℃和Ar下，向(8-((二苯基亞甲基）氨基）-6-(4-甲基吡啶-3-基）噌啉-3-基）氨基甲酸4-硝基苯酯（279 mg，0.482 mmol，粗品）和TEA（98.0mg，0.964mmol）的DCM（10mL）溶液中加入氮雜環丁烷-3-腈鹽酸鹽（57.0mg，0.482mmol）。將混合物在室溫攪拌16h。然後用NaHCO3 溶液（30 mL）淬滅，並用EA（20 mL x 3）萃取。有機層經無水Na2 SO4 乾燥，過濾，濃縮並通過矽膠柱色譜法（DCM：MeOH = 80：1-20：1，v / v）和之後prep-TLC（DCM：MeOH = 20：1）純化，得到為黃色油的3-氰基-N-(8-((二苯基亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-基)氮雜環丁烷-1-甲醯胺（35.0 mg，0.0669 mmol，19.4％產率）。LC-MS（ESI）[M + H]+ 524.2。Step 3. Under 0℃ and Ar, to (8-((diphenylmethylene)amino)-6-(4-methylpyridin-3-yl)cinolin-3-yl)carbamic acid 4- To a solution of nitrophenyl ester (279 mg, 0.482 mmol, crude) and TEA (98.0 mg, 0.964 mmol) in DCM (10 mL) was added azetidine-3-nitrile hydrochloride (57.0 mg, 0.482 mmol). The mixture was stirred at room temperature for 16 h. Then it was quenched with NaHCO 3 solution (30 mL) and extracted with EA (20 mL x 3). The organic layer was dried over anhydrous Na 2 SO 4 , filtered, concentrated and passed through silica gel column chromatography (DCM: MeOH = 80:1-20:1, v/v) and then prep-TLC (DCM: MeOH = 20:1) Purified to obtain 3-cyano-N-(8-((diphenylmethylene)amino)-6-(4-methylpyridin-3-yl)cinolin-3-yl)nitrogen as a yellow oil Etidine-1-carboxamide (35.0 mg, 0.0669 mmol, 19.4% yield). LC-MS (ESI) [M + H] + 524.2.
步驟4.根據通用步驟C 中描述的方法，合成為黃色固體的化合物 85 （8.65mg，0.0241mmol，42.0％產率）。LC-MS (ESI) [M+H]+ 360.4。1 H NMR (400 MHz, DMSO-d6 ): δ 10.1 (s, 1H), 8.46 (d, J = 5.2 Hz, 1H), 8.43 (s, 1H), 8.31 (s, 1H), 7.36 (d, J = 4.8 Hz, 1H), 6.93 (s, 1H), 6.69 (d, J = 1.6 Hz, 1H), 6.63 (s, 2H), 4.39 – 4.34 (m, 2H), 4.24 – 4.20 (m, 2H), 3.81 – 3.77 (m, 1H), 2.31 (s, 3H)。Step 4. According to the method described in general step C , compound 85 (8.65 mg, 0.0241 mmol, 42.0% yield) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 360.4. 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.1 (s, 1H), 8.46 (d, J = 5.2 Hz, 1H), 8.43 (s, 1H), 8.31 (s, 1H), 7.36 (d , J = 4.8 Hz, 1H), 6.93 (s, 1H), 6.69 (d, J = 1.6 Hz, 1H), 6.63 (s, 2H), 4.39 – 4.34 (m, 2H), 4.24 – 4.20 (m, 2H), 3.81 – 3.77 (m, 1H), 2.31 (s, 3H).
N-(8- 氨基 -6-(4- 甲基吡啶 -3- 基 ) 噌啉 -3- 基 )-3-(1- 甲基 -1H- 吡唑 -4- 基 ) 氮雜環丁烷 -1- 甲醯胺（ 86 ）的製備 
步驟1-2。在室溫和Ar下，向活化的Zn（880mg，13.8mmol）在THF（80mL）中的混合物中加入二溴乙烷（200mg，1.06mmol）。將反應混合物加熱至65℃，10分鐘。冷卻至RT後，加入三甲基氯矽烷（115mg，1.06mmol），並在室溫下攪拌1h。然後逐滴加入3-碘氮雜環丁烷-1-羧酸叔丁基酯（3.00g，10.6mmol）的THF（30mL）溶液。將混合物再攪拌1h，然後逐滴加入到在THF（20mL）中的4-碘-1-甲基-1H-吡唑（1.10g，5.29mmol）、CuI（120mg，6.35mmol）和Pd(dppf)Cl2 . DCM（432mg，0.529mmol）的混合物中。將混合物在65℃和Ar下攪拌3h。用水（100mL）稀釋反應混合物，並用EA（30mL×3）萃取。合併的有機層用鹽水（50mL×3）洗滌，用無水Na2 SO4 乾燥，過濾並在減壓下濃縮。殘餘物通過矽膠柱色譜法純化（PE：EA = 20：1-1：1，v，v），然後通過prep-HPLC（NH3 . H2 O）純化，得到為淺色油的3-(1-甲基-1H-吡唑-4-基)氮雜環丁烷-1-羧酸酯（18.0 mg，0.0759 mmol，1.32％收率）。1 H NMR (400 MHz, CDCl3 ): δ 7.46 (s, 1H), 7.39 (s, 1H), 6.29 (d, J = 11.6 Hz, 1H), 5.51 – 5.45 (m, 1H), 4.63 (br s, 1H), 3.98 – 3.97 (m, 2H), 3.95 (s, 3H), 1.44 (s, 9H)。Step 1-2. At room temperature and Ar, to a mixture of activated Zn (880 mg, 13.8 mmol) in THF (80 mL) was added dibromoethane (200 mg, 1.06 mmol). The reaction mixture was heated to 65°C for 10 minutes. After cooling to RT, trimethylchlorosilane (115mg, 1.06mmol) was added and stirred at room temperature for 1h. Then a solution of tert-butyl 3-iodoazetidine-1-carboxylate (3.00 g, 10.6 mmol) in THF (30 mL) was added dropwise. The mixture was stirred for another 1 h, and then added dropwise to 4-iodo-1-methyl-1H-pyrazole (1.10g, 5.29mmol), CuI (120mg, 6.35mmol) and Pd(dppf) in THF (20mL) ) Cl 2. the mixture DCM (432mg, 0.529mmol) in. The mixture was stirred at 65°C under Ar for 3h. The reaction mixture was diluted with water (100 mL) and extracted with EA (30 mL×3). The combined organic layer was washed with brine (50 mL×3), dried over anhydrous Na 2 SO 4 , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE: EA = 20: 1-1 : 1, v, v), and then by prep-HPLC (. NH 3 H 2 O) to give as a pale oil 3- ( 1-Methyl-1H-pyrazol-4-yl)azetidine-1-carboxylate (18.0 mg, 0.0759 mmol, 1.32% yield). 1 H NMR (400 MHz, CDCl 3 ): δ 7.46 (s, 1H), 7.39 (s, 1H), 6.29 (d, J = 11.6 Hz, 1H), 5.51 – 5.45 (m, 1H), 4.63 (br s, 1H), 3.98 – 3.97 (m, 2H), 3.95 (s, 3H), 1.44 (s, 9H).
步驟3.在RT下，在3-(1-甲基-1H-吡唑-4-基)氮雜環丁烷-1-羧酸叔丁基酯（18.0mg，0.0759mmol）的DCM（5mL）的溶液中加入TFA（1mL）。將混合物在RT下攪拌3h。將混合物濃縮並乾燥，將粗品直接用於下一步驟。LC-MS（ESI）[M + H]+ 138.2。Step 3. At RT, in 3-(1-methyl-1H-pyrazol-4-yl)azetidine-1-carboxylic acid tert-butyl ester (18.0mg, 0.0759mmol) in DCM (5mL ) Add TFA (1mL) to the solution. The mixture was stirred at RT for 3h. The mixture was concentrated and dried, and the crude product was used directly in the next step. LC-MS (ESI) [M + H] + 138.2.
步驟4：在0℃和Ar下，在(8-((二苯基亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-基）氨基甲酸4-硝基苯酯(41 mg，0.0766 mmol)和TEA（23.2 mg，0.230mmol）的DCM（5mL）溶液中加入4-(氮雜環丁烷-3-基)-1-甲基-1H-吡唑TFA鹽（18.0mg，0.0766mmol）。將混合物在RT下攪拌16h，然後用NaHCO3 溶液（30mL）稀釋，並用EA（20mL×3）萃取。有機層經無水Na2 SO4 乾燥，過濾並濃縮得到為黃色油的N-(8-((二苯基亞甲基）氨基）-6-(4-甲基吡啶-3-基)噌啉-3-基)-3-(1-甲基-1H-吡唑-4-基)氮雜環丁烷-1-甲醯胺（40.0 mg，粗品，產率100％）。LC-MS（ESI）[M + H]+ 579.6。Step 4: Under 0℃ and Ar, in (8-((diphenylmethylene)amino)-6-(4-methylpyridin-3-yl)cinolin-3-yl)carbamic acid 4- To a solution of nitrophenyl ester (41 mg, 0.0766 mmol) and TEA (23.2 mg, 0.230 mmol) in DCM (5 mL) was added 4-(azetidin-3-yl)-1-methyl-1H-pyridine Azole TFA salt (18.0 mg, 0.0766 mmol). The mixture was stirred at RT for 16 h, then diluted with NaHCO 3 solution (30 mL) and extracted with EA (20 mL×3). The organic layer was dried over anhydrous Na 2 SO 4 , filtered and concentrated to obtain N-(8-((diphenylmethylene)amino)-6-(4-methylpyridin-3-yl)cinnoline as a yellow oil -3-yl)-3-(1-methyl-1H-pyrazol-4-yl)azetidine-1-carboxamide (40.0 mg, crude product, yield 100%). LC-MS (ESI) [M + H] + 579.6.
步驟5.根據通用步驟C 中描述的方法，合成為黃色固體的化合物 86 （1.98mg，0.00478mmol，6.91％產率）。LC-MS (ESI) [M+H]+ 415.5。1 H NMR (400 MHz, DMSO-d6 ): δ 9.88 (s, 1H), 8.46 (d, J = 5.2 Hz, 1H), 8.44 (s, 1H), 8.34 (s, 1H), 7.72 (s, 1H), 7.44 (s, 1H), 7.36 (d, J = 5.2 Hz, 1H) , 6.92 (d, J = 1.6 Hz, 1H) , 6.68 (d, J = 1.6 Hz, 1H), 6.61 (s, 2H), 4.44 – 4.39 (m, 2H), 3.97 – 3.93 (m, 2H), 3.80 (s, 3H), 3.76 – 3.72 (m, 1H), 2.33 (s, 3H)。Step 5. According to the method described in general step C , compound 86 (1.98 mg, 0.00478 mmol, 6.91% yield) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 415.5. 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.88 (s, 1H), 8.46 (d, J = 5.2 Hz, 1H), 8.44 (s, 1H), 8.34 (s, 1H), 7.72 (s , 1H), 7.44 (s, 1H), 7.36 (d, J = 5.2 Hz, 1H), 6.92 (d, J = 1.6 Hz, 1H), 6.68 (d, J = 1.6 Hz, 1H), 6.61 (s , 2H), 4.44 – 4.39 (m, 2H), 3.97 – 3.93 (m, 2H), 3.80 (s, 3H), 3.76 – 3.72 (m, 1H), 2.33 (s, 3H).
(1S,2S)-N-(8- 氨基 -6-((S)-4- 甲基 -2- 氧代惡唑烷 -3- 基 ) 噌啉 -3- 基 )-2-(1- 甲基 -1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺（ 87a ）和 (1R,2R)-N-(8- 氨基 -6-((S)-4- 甲基 -2- 氧代惡唑烷 -3- 基 ) 噌啉 -3 - 基 )-2-(1- 甲基 -1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺（ 87b ）的製備 
步驟1。在室溫下，向(反式)-N-(6-氯-8-((二苯基亞甲基)氨基)噌啉-3-基)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（750 mg，1.48 mmol）和(S)-4-甲基惡唑烷-2-酮（300 mg，2.97 mmol）的二氧雜環己烷（6.00 mL）的溶液中加入4,5-雙(二苯基膦)-9,9-二甲基氧雜蒽（342mg，0.591mmol）、磷酸鉀（942mg，4.44mmol）和Pd2 (dba)3 （276mg，0.300mmol）。將混合物在密閉管中於110°C攪拌3小時。將混合物冷卻至室溫。過濾混合物，濾餅用乙酸乙酯（20.0mL X 2）洗滌。用水（40.0mL）稀釋濾液，用乙酸乙酯（20.0mL X 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓下濃縮。殘餘物通過柱色譜法（二氯甲烷/甲醇= 20/1）純化，得到為黃色固體的(反式)-N-(8-((二苯基亞甲基)氨基)-6-((S)-4-甲基-2-氧代惡唑烷-3-基)噌啉-3-基)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（400 mg，47.3％產率）。LC-MS（ESI）[M + H]+ 572.1。step 1. At room temperature, to (trans)-N-(6-chloro-8-((diphenylmethylene)amino)cinnolin-3-yl)-2-(1-methyl-1H-pyridine) (Azol-4-yl)cyclopropane-1-carboxamide (750 mg, 1.48 mmol) and (S)-4-methyloxazolidin-2-one (300 mg, 2.97 mmol) in dioxane Add 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (342mg, 0.591mmol), potassium phosphate (942mg, 4.44mmol) and Pd 2 ( dba) 3 (276 mg, 0.300 mmol). The mixture was stirred in a closed tube at 110°C for 3 hours. The mixture was cooled to room temperature. The mixture was filtered, and the filter cake was washed with ethyl acetate (20.0 mL X 2). The filtrate was diluted with water (40.0 mL) and extracted with ethyl acetate (20.0 mL X 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane/methanol=20/1) to obtain (trans)-N-(8-((diphenylmethylene)amino)-6-(( S)-4-Methyl-2-oxazolidin-3-yl)cinnoline-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1- Formamide (400 mg, 47.3% yield). LC-MS (ESI) [M + H] + 572.1.
步驟2.根據通用步驟C 中所述的方法，合成為黃色固體的(反式)-N-(8-氨基-6-((S)-4-甲基-2-氧代惡唑烷-3-基)噌啉-3-基)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（100mg，35.1％產率）。LC-MS（ESI）[M + H]+ 408.1。Step 2. According to the method described in general step C , synthesize (trans)-N-(8-amino-6-((S)-4-methyl-2-oxooxazolidine-) as a yellow solid 3-yl)Cinolin-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-carboxamide (100 mg, 35.1% yield). LC-MS (ESI) [M + H] + 408.1.
步驟3. 利用SFC（系統：Waters SFC 80；色譜柱名稱：CHIRALPAK®OJ-H；色譜柱尺寸：250*25mm 10 μm）；流動相A：超臨界CO2 ；流動相B：MeOH（0.1％NH3 ∙H2 O）；A：B：60:40；波長：214nm；流速：70 mL / min；柱溫：RT；柱壓：100bar；進樣：3.5mL；迴圈時間：5min；樣品溶液的製備：將樣品溶於約23mL的MeOH中）分離（反式）-N-（8-氨基-6-（（S）-4-甲基-2-氧代惡唑烷-3-基）噌啉-3-基）-2-（1-甲基-1H-吡唑-4-基）環丙烷-1-甲醯胺（100 mg，0.245 mmol）固體，得到呈黃色固體的(1S, 2S)-N-(8-氨基-6-((S)-4-甲基-2-氧雜惡唑烷-3-基)噌啉-3-基)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（27.09 mg，P1，117）和為黃色固體的( 1R，2R)-N-(8-氨基-6-((S)-4-甲基-2-氧雜惡唑烷-3-基)噌啉-3-基)-2-(1-甲基-1H-吡唑-4 –基)環丙烷-1-甲醯胺（35.37 mg，P2，118）。Step 3. Use SFC (system: Waters SFC 80; column name: CHIRALPAK®OJ-H; column size: 250*25mm 10 μm); mobile phase A: supercritical CO 2 ; mobile phase B: MeOH (0.1% NH 3 ∙H 2 O); A: B: 60:40; wavelength: 214nm; flow rate: 70 mL/min; column temperature: RT; column pressure: 100 bar; injection: 3.5 mL; loop time: 5 min; sample Solution preparation: Dissolve the sample in about 23mL of MeOH) to separate (trans)-N-(8-amino-6-((S)-4-methyl-2-oxooxazolidin-3-yl) ) Cinolin-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-carboxamide (100 mg, 0.245 mmol) as a solid, and (1S , 2S)-N-(8-amino-6-((S)-4-methyl-2-oxazolidine-3-yl)cinolin-3-yl)-2-(1-methyl -1H-pyrazol-4-yl)cyclopropane-1-carboxamide (27.09 mg, P1,117) and (1R, 2R)-N-(8-amino-6-((S)) as a yellow solid -4-Methyl-2-oxazolidine-3-yl)cinnoline-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-methanone Amine (35.37 mg, P2, 118).
(1S,2S)-N-(8-氨基-6-((S)-4-甲基-2-氧代惡唑烷-3-基)噌啉-3-基)-2-(1-甲基-1H-吡唑- 4-基)環丙烷-1-甲醯胺（P1，87a ），LC-MS (ESI) [M+H]+ 408.0。SFC 方法注釋：CO2 /MeOH/DEA 70/30/0.03 1.8mL/min OJ, 3um, 3*100 (Daicel). RT = 1.826 min, e.e% =99.3%.1 H NMR (400 MHz, DMSO-d6 ): δ 11.41 (s, 1H), 8.45 (s, 1H), 7.58 (s, 1H), 7.31 (s, 1H), 7.24 (d, J = 2.4 Hz, 1H), 6.94 (d, J = 2.4Hz, 1H), 6.68 (s, 2H), 4.74 - 4.70 (m, 1H), 4.59 - 4.55 (m, 1H), 4.09 - 4.06 (m, 1H), 3.78 (s, 3H), 2.29 - 2.23 (m, 2H), 1.44 - 1.39 (m, 1H), 1.32 (d, J = 6.4 Hz, 3H), 1.25 - 1.20 (m, 1H)。(1S,2S)-N-(8-amino-6-((S)-4-methyl-2-oxooxazolidin-3-yl) cinnolin-3-yl)-2-(1- Methyl-1H-pyrazole-4-yl)cyclopropane-1-carboxamide (P1, 87a ), LC-MS (ESI) [M+H] + 408.0. SFC method note: CO 2 /MeOH/DEA 70/30/0.03 1.8mL/min OJ, 3um, 3*100 (Daicel). RT = 1.826 min, ee% =99.3%. 1 H NMR (400 MHz, DMSO- d 6 ): δ 11.41 (s, 1H), 8.45 (s, 1H), 7.58 (s, 1H), 7.31 (s, 1H), 7.24 (d, J = 2.4 Hz, 1H), 6.94 (d, J = 2.4Hz, 1H), 6.68 (s, 2H), 4.74-4.70 (m, 1H), 4.59-4.55 (m, 1H), 4.09-4.06 (m, 1H), 3.78 (s, 3H), 2.29- 2.23 (m, 2H), 1.44-1.39 (m, 1H), 1.32 (d, J = 6.4 Hz, 3H), 1.25-1.20 (m, 1H).
(1R,2R)-N-(8-氨基-6-((S)-4-甲基-2-氧代惡唑烷-3-基)噌啉-3-基)-2-(1-甲基-1H-吡唑- 4-基)環丙烷-1-甲醯胺（P2，87b ）。LC-MS (ESI) [M+H]+ 408.0。SFC方法注釋：CO2 /MeOH/DEA 70/30/0.03 1.8mL/min OJ，3um， 3*100 (Daicel)。RT = 2.569 min， e.e% =99.28%.1 H NMR (400 MHz, DMSO-d6 ): δ 11.39 (s, 1H), 8.43 (s, 1H), 7.56 (s, 1H), 7.29 (s, 1H), 7.23 (d, J = 2.4 Hz, 1H), 6.92 (d, J = 2.4 Hz, 1H), 6.67 (s, 2H), 4.73 - 4.68 (m, 1H), 4.57 - 4.53 (m, 1H), 4.07 - 4.04 (m, 1H), 3.76 (s, 3H), 2.29 - 2.19 (m, 2H), 1.43 - 1.38 (m, 1H), 1.31 (d, J = 6.0 Hz, 3H), 1.23 - 1.18 (m, 1H)。(1R,2R)-N-(8-amino-6-((S)-4-methyl-2-oxooxazolidin-3-yl)cinolin-3-yl)-2-(1- Methyl-1H-pyrazole-4-yl)cyclopropane-1-carboxamide (P2, 87b ). LC-MS (ESI) [M+H] + 408.0. SFC method note: CO 2 /MeOH/DEA 70/30/0.03 1.8mL/min OJ, 3um, 3*100 (Daicel). RT = 2.569 min, ee% =99.28%. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.39 (s, 1H), 8.43 (s, 1H), 7.56 (s, 1H), 7.29 (s, 1H), 7.23 (d, J = 2.4 Hz, 1H), 6.92 (d, J = 2.4 Hz, 1H), 6.67 (s, 2H), 4.73-4.68 (m, 1H), 4.57-4.53 (m, 1H ), 4.07-4.04 (m, 1H), 3.76 (s, 3H), 2.29-2.19 (m, 2H), 1.43-1.38 (m, 1H), 1.31 (d, J = 6.0 Hz, 3H), 1.23- 1.18 (m, 1H).
(1R,2R,3R)-N-(8- 氨基 -6-((S)-4- 甲基 -2- 氧代惡唑烷 -3- 基 ) 噌啉 -3- 基 )-2- 甲基 -3-( 1- 甲基 -1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺（ 88a ）， (1S,2S,3S)-N-(8- 氨基 -6-((S)-4- 甲基 -2- 氧代惡唑烷 -3- 基 ) 噌啉 -3- 基 )-2- 甲基 -3-(1- 甲基 -1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺（ 88b ），（ 1S,2R,3S)-N- (8- 氨基 -6-((S)-4- 甲基 -2- 氧代惡唑烷 -3- 基 ) 噌啉 -3- 基 )-2- 甲基 -3-(1- 甲基 -1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺（ 88c ）和（ 1R,2S,3R)-N-(8- 氨基 -6-((S)-4- 甲基 -2- 氧代惡唑烷 -3- 基 ) 噌啉 -3- 基 )- 2- 甲基 -3-(1- 甲基 -1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺（ 88d ） 
步驟1.在(反式)-2-甲基-3-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲酸（210 mg，1.16 mmol，根據WO2018183964製備）、6-氯-8-(((二苯基亞甲基)氨基)噌啉-3-胺（418mg，1.16mmol）的DCM（25mL）溶液中加入吡啶（3.60mL）。在室溫下攪拌30min後，在0℃下添加POCl3 （508mg，3.32mmol）的DCM（3.00mL）溶液，並在室溫下攪拌20分鐘。將混合物用飽和NaHCO3 稀釋，並用EA（10.0 mL x 3）萃取。合併的有機相經Na2 SO4 乾燥並真空濃縮得到殘餘物，將其通過prep-TLC（PE：EA=2：1）純化，得到為黃色固體的標題化合物（260mg，收率43.1％）。LC-MS（ESI）[M + H]+ 521.0。Step 1. In (trans)-2-methyl-3-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-carboxylic acid (210 mg, 1.16 mmol, prepared according to WO2018183964), 6 -Chloro-8-(((diphenylmethylene)amino)cinnoline-3-amine (418mg, 1.16mmol) in DCM (25mL) was added with pyridine (3.60mL). After stirring for 30min at room temperature Add a solution of POCl 3 (508 mg, 3.32 mmol) in DCM (3.00 mL) at 0°C and stir at room temperature for 20 minutes. The mixture was diluted with saturated NaHCO 3 and extracted with EA (10.0 mL x 3). Combine The organic phase was dried over Na 2 SO 4 and concentrated in vacuo to obtain a residue, which was purified by prep-TLC (PE:EA=2:1) to obtain the title compound (260 mg, yield 43.1%) as a yellow solid. -MS (ESI) [M + H] + 521.0.
步驟2.以與製備化合物 87a / b 類似的方法，合成為黃色固體的(反式)-N-(8-(((二苯基亞甲基)氨基)-6-((S)-4-甲基-2-氧代惡唑烷-3-基)噌啉- 3-基)-2-甲基-3-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（150mg，收率89.3％）。LC-MS（ESI）[M + H]+ 586.2。Step 2. Using a method similar to the preparation of compound 87a/b , synthesize (trans)-N-(8-(((diphenylmethylene)amino)-6-((S)-4 as a yellow solid) -Methyl-2-oxooxazolidin-3-yl)cinnoline-3-yl)-2-methyl-3-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1 -Formamide (150 mg, yield 89.3%). LC-MS (ESI) [M + H] + 586.2.
步驟3.根據通用步驟C 中所述的方法，合成為黃色固體的外消旋體 88 （100mg，產率92.7％）。LC-MS（ESI）[M + H]+ 422.0。Step 3. According to the method described in General Step C , the racemate 88 (100 mg, yield 92.7%) was synthesized as a yellow solid. LC-MS (ESI) [M + H] + 422.0.
利用SFC（ChiralPak AD-H Daicel Chemical Industries，Ltd，IC 3.0×25cm，5um，A：超臨界CO2 ，B：EtOH；A：B = 60/40；B：EtOH；A：B = 60/40；B：EtOH；A：B = 60/40；A：B = 60/40；A：B = 60/40；A：B = 60/40；流量以50mL / min表示；色譜柱溫度（T）以38℃表示；背壓（BPR）以100bars表示）分離消旋體88 （100mg，0.238mmol），得到88a （7.00 mg，RT = 1.560 min，ee = 100.00％，7.00％產率）、88b （22.0 mg，RT = 1.682min，ee = 100.00％，22.0％產率）、88c （4.00 mg，RT = 1.882 min，ee = 95.30％，4.00％產率）、88d（14.0 mg，RT = 2.378 min，ee = 99.68％，14.0％ 產率）。Using SFC (ChiralPak AD-H Daicel Chemical Industries, Ltd, IC 3.0×25cm, 5um, A: supercritical CO 2 , B: EtOH; A: B = 60/40; B: EtOH; A: B = 60/40 ; B: EtOH; A: B = 60/40; A: B = 60/40; A: B = 60/40; A: B = 60/40; the flow rate is expressed by 50 mL/min; the column temperature (T) Expressed at 38°C; back pressure (BPR) expressed at 100bars) Separate racemate 88 (100mg, 0.238mmol) to obtain 88a (7.00 mg, R T = 1.560 min, ee = 100.00%, 7.00% yield), 88b (22.0 mg, R T = 1.682min, ee = 100.00%, 22.0% yield), 88c (4.00 mg, R T = 1.882 min, ee = 95.30%, 4.00% yield), 88d (14.0 mg, R T = 2.378 min, ee = 99.68%, 14.0% yield).
88a : LC-MS (ESI) [M+H]+ 422.0。1 H NMR (400 MHz, DMSO-d6 ): δ 11.35 (s, 1H), 8.48 (s, 1H), 7.54 (s, 1H), 7.26 (s, 2H ), 6.94 (s, 1H), 6.67 (s, 2H), 4.74 – 4.70 (m ,1H), 4.59 – 4.51 (m, 1H), 4.10 – 4.05 (m, 1H), 3.77 (s, 3H), 2.33 – 2.28 (m, 1H), 2.21 – 2.18 (m, 1H), 1.63 – 1.60 (m, 1H), 1.33 – 1.26 (m, 6H). 88a : LC-MS (ESI) [M+H] + 422.0. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.35 (s, 1H), 8.48 (s, 1H), 7.54 (s, 1H), 7.26 (s, 2H ), 6.94 (s, 1H), 6.67 (s, 2H), 4.74 – 4.70 (m ,1H), 4.59 – 4.51 (m, 1H), 4.10 – 4.05 (m, 1H), 3.77 (s, 3H), 2.33 – 2.28 (m, 1H), 2.21 – 2.18 (m, 1H), 1.63 – 1.60 (m, 1H), 1.33 – 1.26 (m, 6H).
88b : LC-MS (ESI) [M+H]+ 422.0。1 H NMR (400 MHz, DMSO-d6 ): δ 11.36 (s, 1H), 8.45(s, 1H), 7.55 (s, 1H), 7.30 (s, 1H ), 7.24 (s, 1H ), 6.93 (s, 1H), 6.70 (s, 2H), 4.74 – 4.70 (m ,1H), 4.59 – 4.51 (m, 1H), 4.10 – 4.05 (m, 1H), 3.81 (s, 3H), 2.36 – 2.30 (m, 1H), 2.25 – 2.18 (m, 1H), 1.63 – 1.60 (m, 1H), 1.33 – 1.31 (m, 3H), 0.98 – 0.96 (m, 3H). 88b : LC-MS (ESI) [M+H] + 422.0. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.36 (s, 1H), 8.45 (s, 1H), 7.55 (s, 1H), 7.30 (s, 1H ), 7.24 (s, 1H ), 6.93 (s, 1H), 6.70 (s, 2H), 4.74 – 4.70 (m ,1H), 4.59 – 4.51 (m, 1H), 4.10 – 4.05 (m, 1H), 3.81 (s, 3H), 2.36 – 2.30 (m, 1H), 2.25 – 2.18 (m, 1H), 1.63 – 1.60 (m, 1H), 1.33 – 1.31 (m, 3H), 0.98 – 0.96 (m, 3H).
88c : LC-MS (ESI) [M+H]+ 422.0。1 H NMR (400 MHz, DMSO-d6 ): δ 11.33 (s, 1H), 8.47 (s, 1H), 7.54 (s, 1H), 7.26 (s, 2H ), 6.94 (s, 1H), 6.67 (s, 2H), 4.73 – 4.70 (m ,1H), 4.59 – 4.51 (m, 1H), 4.10 – 4.05 (m, 1H), 3.77 (s, 3H), 2.35 – 2.28 (m, 1H), 2.21 – 2.17 (m, 1H), 1.63 – 1.58 (m, 1H), 1.33 – 1.23 (m, 6H). 88c : LC-MS (ESI) [M+H] + 422.0. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.33 (s, 1H), 8.47 (s, 1H), 7.54 (s, 1H), 7.26 (s, 2H ), 6.94 (s, 1H), 6.67 (s, 2H), 4.73 – 4.70 (m ,1H), 4.59 – 4.51 (m, 1H), 4.10 – 4.05 (m, 1H), 3.77 (s, 3H), 2.35 – 2.28 (m, 1H), 2.21 – 2.17 (m, 1H), 1.63 – 1.58 (m, 1H), 1.33 – 1.23 (m, 6H).
88d : LC-MS (ESI) [M+H]+ 422.0。1 H NMR (400 MHz, DMSO-d6 ): δ 11.37 (s, 1H), 8.45 (s, 1H), 7.56 (s, 1H), 7.31 (s, 1H ), 7.24 (s, 1H ), 6.93 (s, 1H), 6.70 (s, 2H), 4.73 – 4.70 (m ,1H), 4.59 – 4.51 (m, 1H), 4.10 – 4.05 (m, 1H), 3.77 (s, 3H), 2.38 – 2.30 (m, 1H), 2.24 – 2.18 (m, 1H), 1.65 – 1.62 (m, 1H), 1.33 – 1.30 (m, 3H), 0.98 – .096 (m, 3H). 88d : LC-MS (ESI) [M+H] + 422.0. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.37 (s, 1H), 8.45 (s, 1H), 7.56 (s, 1H), 7.31 (s, 1H ), 7.24 (s, 1H ), 6.93 (s, 1H), 6.70 (s, 2H), 4.73 – 4.70 (m ,1H), 4.59 – 4.51 (m, 1H), 4.10 – 4.05 (m, 1H), 3.77 (s, 3H), 2.38 – 2.30 (m, 1H), 2.24 – 2.18 (m, 1H), 1.65 – 1.62 (m, 1H), 1.33 – 1.30 (m, 3H), 0.98 – .096 (m, 3H).
與製備化合物And preparation compound 87a / b87a / b 類似的方法製備化合物Similar methods to prepare compounds 89-9389-93 。. ( ( 反式Trans )-N-(8-)-N-(8- 氨基Amino -6-((S)-4--6-((S)-4- 甲基methyl -2--2- 氧代惡唑烷Oxoxazolidine -3--3- 基base )) 噌啉Cinolin -3--3- 基base )-2-(1-)-2-(1- 乙基Ethyl -1H--1H- 吡唑Pyrazole -4--4- 基base )) 環丙烷Cyclopropane -1--1- 甲醯胺（Formamide ( 8989 ）)
LC-MS (ESI) [M+H]+ 422.1。1 H NMR (400 MHz, DMSO-d6 ): δ 11.40 (s, 1H), 8.45 (s, 1H), 7.64 (s, 1H), 7.32 (s, 1H), 7.24 (s, 1H), 6.94 (d, J = 2.0 Hz, 1H), 6.69 (s, 2H), 4.73 - 4.70 (m, 1H), 4.59 - 4.55 (m, 1H), 4.09 - 4.03 (m, 3H), 2.29 - 2.22 (m, 2H), 1.44 - 1.42 (m, 1H), 1.36 - 1.31 (m, 6H), 1.30 - 1.23(m, 1H).LC-MS (ESI) [M+H] + 422.1. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.40 (s, 1H), 8.45 (s, 1H), 7.64 (s, 1H), 7.32 (s, 1H), 7.24 (s, 1H), 6.94 (d, J = 2.0 Hz, 1H), 6.69 (s, 2H), 4.73-4.70 (m, 1H), 4.59-4.55 (m, 1H), 4.09-4.03 (m, 3H), 2.29-2.22 (m , 2H), 1.44-1.42 (m, 1H), 1.36-1.31 (m, 6H), 1.30-1.23 (m, 1H).
(( 反式Trans )-N-(8-)-N-(8- 氨基Amino -6-((S)-4--6-((S)-4- 甲基methyl -2--2- 氧代惡唑烷Oxoxazolidine -3--3- 基base )) 噌啉Cinolin -3--3- 基base )-2-(1-()-2-(1-( 氧雜Oxa -3--3- 基base )- 1H-)- 1H- 吡唑Pyrazole -4--4- 基base )) 環丙烷Cyclopropane -1--1- 甲醯胺（Formamide ( 9090 ）)
LC-MS (ESI) [M+H]+ 450.2。1 H NMR (400 MHz, DMSO-d6 ): δ 11.41 (s, 1H), 8.45 (s, 1H), 7.82 (s, 1H), 7.49 (s, 1H), 7.24 – 7.23 (m, 1H), 6.94 – 6.93 (m, 1H), 6.68 (s, 2H), 5.52 – 5.49 (m, 1H), 4.83 – 4.92 (m, 4H), 4.74 – 4.72 (m, 1H), 4.57 (t, J = 8.0 Hz, 1H), 4.09 – 4.05 (m, 1H), 2.30 – 2.23 (m, 2H), 1.46 – 1.41 (m, 1H), 1.46 – 1.40 (m, 1H), 1.32 (d, J = 6.0 Hz, 3H), 1.29 – 1.23 (m, 1H)。LC-MS (ESI) [M+H] + 450.2. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.41 (s, 1H), 8.45 (s, 1H), 7.82 (s, 1H), 7.49 (s, 1H), 7.24 – 7.23 (m, 1H) , 6.94 – 6.93 (m, 1H), 6.68 (s, 2H), 5.52 – 5.49 (m, 1H), 4.83 – 4.92 (m, 4H), 4.74 – 4.72 (m, 1H), 4.57 (t, J = 8.0 Hz, 1H), 4.09 – 4.05 (m, 1H), 2.30 – 2.23 (m, 2H), 1.46 – 1.41 (m, 1H), 1.46 – 1.40 (m, 1H), 1.32 (d, J = 6.0 Hz , 3H), 1.29 – 1.23 (m, 1H).
(( 反式Trans )-N-(8-)-N-(8- 氨基Amino -6-((S)-4--6-((S)-4- 甲基methyl -2--2- 氧代惡唑烷Oxoxazolidine -3--3- 基base )) 噌啉Cinolin -3--3- 基base )-2-(1-(2,2,2-)-2-(1-(2,2,2- 三氟乙基Trifluoroethyl )-1H-)-1H- 吡唑Pyrazole -4--4- 基base )) 環丙烷Cyclopropane -1--1- 甲醯胺（Formamide ( 9191 ）)
LC-MS (ESI) [M+H]+ 476.0。1 H NMR (400 MHz, DMSO-d6 ): δ 11.44 (s, 1H), 8.46 (d, J = 4.8 Hz, 1H), 7.75 (d, J = 5.2 Hz, 1H), 7.50 (d, J = 5.2 Hz, 1H), 7.25 (s, 1H), 6.94 (s, 1H), 6.70 (s, 2H), 5.12 - 5.03 (m, 2H), 4.77 - 4.71 (m, 1H), 4.62 - 4.55 (m, 1H), 4.11 - 4.06 (m, 1H), 2.38 - 2.26 (m, 2H), 1.49 - 1.42 (m, 1H), 1.36 - 1.24 (m, 4H)。LC-MS (ESI) [M+H] + 476.0. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.44 (s, 1H), 8.46 (d, J = 4.8 Hz, 1H), 7.75 (d, J = 5.2 Hz, 1H), 7.50 (d, J = 5.2 Hz, 1H), 7.25 (s, 1H), 6.94 (s, 1H), 6.70 (s, 2H), 5.12-5.03 (m, 2H), 4.77-4.71 (m, 1H), 4.62-4.55 ( m, 1H), 4.11-4.06 (m, 1H), 2.38-2.26 (m, 2H), 1.49-1.42 (m, 1H), 1.36-1.24 (m, 4H).
(( 反式Trans )-N-(8-)-N-(8- 氨基Amino -6-((S)-4--6-((S)-4- 甲基methyl -2--2- 氧代惡唑烷Oxoxazolidine -3--3- 基base )) 噌啉Cinolin -3--3- 基base )-2-(1-)-2-(1- 異丙基Isopropyl -1H--1H- 吡唑Pyrazole -4--4- 基base )) 環丙胺Cyclopropylamine -1--1- 甲醯胺（Formamide ( 9292 ）)
LC-MS (ESI) [M+H]+ 436.5。1 H NMR (400 MHz, DMSO-d6 ): δ 11.39 (s, 1H), 8.45 (s, 1H), 7.68 (s, 1H), 7.31 (s, 1H), 7.24 (s, 1H), 6.94 (s, 1H), 6.69 (s, 2H), 4.73 - 4.70 (m, 1H), 4.59 - 4.55 (m, 1H), 4.43 - 4.39 (m, 1H), 4.09 - 4.06 (m, 1H), 2.44 - 2.42 (m, 2H), 1.43 – 1.24 (m, 11H)。LC-MS (ESI) [M+H] + 436.5. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.39 (s, 1H), 8.45 (s, 1H), 7.68 (s, 1H), 7.31 (s, 1H), 7.24 (s, 1H), 6.94 (s, 1H), 6.69 (s, 2H), 4.73-4.70 (m, 1H), 4.59-4.55 (m, 1H), 4.43-4.39 (m, 1H), 4.09-4.06 (m, 1H), 2.44 -2.42 (m, 2H), 1.43 – 1.24 (m, 11H).
(( 反式Trans )-N-(8-)-N-(8- 氨基Amino -6-((S)-4--6-((S)-4- 甲基methyl -2--2- 氧代惡唑烷Oxoxazolidine -3--3- 基base )) 噌啉Cinolin -3--3- 基base )-2-(1-(2-)-2-(1-(2- 甲氧基乙基Methoxyethyl )-1H-)-1H- 吡唑Pyrazole -4--4- 基base )) 環丙烷Cyclopropane -1--1- 甲醯胺（Formamide ( 9393 ）)
LC-MS (ESI) [M+H]+ 452.1.1 H NMR (400 MHz, DMSO-d6 ): δ 11.41 (s, 1H), 8.45 (s, 1H), 7.60 (s, 1H), 7.33 (s, 1H), 7.24 – 7.23 (m, 1H), 6.94 – 6.93 (m, 1H), 6.68 (s, 2H), 4.74 - 4.70 (m, 1H), 4.57 (t, J = 6.0 Hz, 1H), 4.18 (t, J = 6.0 Hz, 2H), 4.09 - 4.06 (m, 1H), 3.65 (t, J = 5.2 Hz, 2H), 3.23 (s, 3H), 2.32 – 2.22 (m, 2H), 1.45 – 1.40 (m, 1H), 1.32 (d, J = 6.4 Hz, 3H), 1.25 – 1.21 (m, 1H)。LC-MS (ESI) [M+H] + 452.1. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.41 (s, 1H), 8.45 (s, 1H), 7.60 (s, 1H), 7.33 (s, 1H), 7.24 – 7.23 (m, 1H), 6.94 – 6.93 (m, 1H), 6.68 (s, 2H), 4.74-4.70 (m, 1H), 4.57 (t, J = 6.0 Hz, 1H ), 4.18 (t, J = 6.0 Hz, 2H), 4.09-4.06 (m, 1H), 3.65 (t, J = 5.2 Hz, 2H), 3.23 (s, 3H), 2.32-2.22 (m, 2H) , 1.45 – 1.40 (m, 1H), 1.32 (d, J = 6.4 Hz, 3H), 1.25 – 1.21 (m, 1H).
( 反式 )-N-(8- 氨基 -6-((S)-4- 甲基 -2- 氧代惡唑烷 -3- 基 ) 噌啉 -3- 基 )-2-(1- 環丙基 -1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺（ 94 ） 
步驟1. 在O2 氣球下，將（反式）-2-（1H-吡唑-4-基）環丙烷-1-羧酸乙酯（180 mg，1.00 mmol）、環丙基硼酸（172 mg，2.00 mmol）、碳酸鈉（212 mg，2.00 mmol）、Cu(OAc)2 （181 mg，1.00 mmol）和2,2-聯吡啶（156 mg，1.00 mmol）的DCE（10.0 mL）溶液在 80 ℃下攪拌16小時。將混合物用水（50.0mL）稀釋，然後用EtOAc（20.0mL×3）萃取。合併的有機層經無水硫酸鈉乾燥，過濾並濃縮得到粗產物。殘餘物通過柱色譜法（矽膠，石油醚/乙酸乙酯= 3/1）純化，得到為棕色油的（反式）-2-（1-環丙基-1H-吡唑-4-基）環丙烷-1-羧酸乙酯（81.0mg，收率36.7％）。LC-MS（ESI）[M + H]+ 221.1。Step 1. Under the O 2 balloon, add (trans)-2-(1H-pyrazol-4-yl)cyclopropane-1-carboxylic acid ethyl ester (180 mg, 1.00 mmol), cyclopropylboronic acid (172 mg, 2.00 mmol), sodium carbonate (212 mg, 2.00 mmol), Cu(OAc) 2 (181 mg, 1.00 mmol) and 2,2-bipyridine (156 mg, 1.00 mmol) in DCE (10.0 mL) solution Stir at 80°C for 16 hours. The mixture was diluted with water (50.0 mL), and then extracted with EtOAc (20.0 mL×3). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The residue was purified by column chromatography (silica gel, petroleum ether/ethyl acetate = 3/1) to obtain (trans)-2-(1-cyclopropyl-1H-pyrazol-4-yl) as a brown oil Ethyl cyclopropane-1-carboxylate (81.0 mg, yield 36.7%). LC-MS (ESI) [M + H] + 221.1.
步驟2.將（反式）-2-（1-環丙基-1H-吡唑-4-基）環丙烷-1-羧酸乙酯用LiOH在MeOH / H2 O中水解，得到為棕色油的(反式)-2-（1-環丙基-1H -吡唑-4-基）環丙烷-1-羧酸（70.0 mg，89.1％產率）。LC-MS（ESI）[M + MeCN]+ 234.2。Step 2. Hydrolyze (trans)-2-(1-cyclopropyl-1H-pyrazol-4-yl)cyclopropane-1-carboxylic acid ethyl ester with LiOH in MeOH/H 2 O to obtain brown Oily (trans)-2-(1-cyclopropyl-1H-pyrazol-4-yl)cyclopropane-1-carboxylic acid (70.0 mg, 89.1% yield). LC-MS (ESI) [M + MeCN] + 234.2.
步驟3-5。以製備化合物 87a / b 類似的方法製備化合物 94 （2.95mg，33.9％產率）。LC-MS (ESI) [M+H]+ 434.5.1 H NMR (400 MHz, DMSO-d6 ) δ 11.39 (s, 1H), 8.45 (s, 1H), 7.68 (s, 1H), 7.31 (s, 1H), 7.24 (s, 1H), 6.94 (s, 1H), 6.69 (s, 2H), 4.73 – 4.72 (m, 1H), 4.59 – 4.55 (m, 1H), 4.09 – 4.06 (m, 1H), 3.64 – 3.62 (m, 1H), 2.28 – 2.22 (m, 2H), 1.42 – 1.41 (m, 1H), 1.33 – 1.24 (m, 4H), 0.97 – 0.92 (m, 4H)。Steps 3-5. Compound 94 (2.95 mg, 33.9% yield) was prepared in a similar way to the preparation of compound 87a/b. LC-MS (ESI) [M+H] + 434.5. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.39 (s, 1H), 8.45 (s, 1H), 7.68 (s, 1H), 7.31 ( s, 1H), 7.24 (s, 1H), 6.94 (s, 1H), 6.69 (s, 2H), 4.73 – 4.72 (m, 1H), 4.59 – 4.55 (m, 1H), 4.09 – 4.06 (m, 1H), 3.64 – 3.62 (m, 1H), 2.28 – 2.22 (m, 2H), 1.42 – 1.41 (m, 1H), 1.33 – 1.24 (m, 4H), 0.97 – 0.92 (m, 4H).
( 反式 )-N-(8- 氨基 -6-((S)-4- 甲基 -2- 氧代惡唑烷 -3- 基 ) 噌啉 -3- 基 )-2-(1,5- 二甲基 -1H- 吡唑 - 4- 基 ) 環丙烷 -1- 甲醯胺（ 95 ） 
步驟1。用LiOH在MeOH / H2 O中水解(反式)-2-(1,5-二甲基-1H-吡唑-4-基)環丙烷-1-甲酸乙酯，得到為棕色油的(反式)-2-(1,5 --二甲基-1H-吡唑-4-基)環丙烷-1-甲酸（167 mg，42.9％產率）。LC-MS（ESI）[M + H]+ 181.2。step 1. Hydrolyze (trans)-2-(1,5-dimethyl-1H-pyrazol-4-yl)cyclopropane-1-carboxylic acid ethyl ester with LiOH in MeOH/H 2 O to obtain ( Trans)-2-(1,5-Dimethyl-1H-pyrazol-4-yl)cyclopropane-1-carboxylic acid (167 mg, 42.9% yield). LC-MS (ESI) [M + H] + 181.2.
步驟3-5。以製備化合物 87a / b 類似的方法製備黃色固體化合物 95 （3.06mg，12.2％產率）。LC-MS (ESI) [M+H]+ 422.4.1 H NMR (400 MHz, DMSO-d6 ): δ 11.45 (s, 1H), 8.47 (s, 1H), 7.25 (d, J = 1.6 Hz, 1H), 7.13 (s, 1H), 6.93 (d, J = 2.0 Hz, 1H), 6.69 (s, 2H), 4.73 – 4.70 (m, 1H), 4.57 (t, J = 8.4 Hz,1H), 4.09 – 4.06 (m, 1H), 3.69 (s, 3H), 2.29 – 2.26 (m, 1H), 2.23 (s, 3H), 2.13 – 2.09 (m, 1H), 1.42 – 1.39 (m, 1H), 1.32 (d, J = 6.0 Hz, 3H), 1.18 – 1.17 (m, 1H)。Steps 3-5. A yellow solid compound 95 (3.06 mg, 12.2% yield) was prepared in a similar way to the preparation of compound 87a/b. LC-MS (ESI) [M+H] + 422.4. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.45 (s, 1H), 8.47 (s, 1H), 7.25 (d, J = 1.6 Hz , 1H), 7.13 (s, 1H), 6.93 (d, J = 2.0 Hz, 1H), 6.69 (s, 2H), 4.73 – 4.70 (m, 1H), 4.57 (t, J = 8.4 Hz, 1H) , 4.09 – 4.06 (m, 1H), 3.69 (s, 3H), 2.29 – 2.26 (m, 1H), 2.23 (s, 3H), 2.13 – 2.09 (m, 1H), 1.42 – 1.39 (m, 1H) , 1.32 (d, J = 6.0 Hz, 3H), 1.18 – 1.17 (m, 1H).
(1R,2R)-N-(8-(1R,2R)-N-(8- 氨基Amino -6-((S)-4--6-((S)-4- 乙基Ethyl -2--2- 氧代惡唑烷Oxoxazolidine -3--3- 基base )) 噌啉Cinolin -3--3- 基base )-2-(1-)-2-(1- 甲基methyl -1H--1H- 吡唑Pyrazole - 4--4- 基base )) 環丙烷Cyclopropane -1--1- 羧醯胺（Carboxamide ( 96a96a ）和（)with( 1S,2S)-N-(8-1S,2S)-N-(8- 氨基Amino -6-((S)-4--6-((S)-4- 乙基Ethyl -2--2- 氧代惡唑烷Oxoxazolidine -3--3- 基base )) 噌啉Cinolin -3--3- 基base )-2-(1-)-2-(1- 甲基methyl -1H--1H- 吡唑Pyrazole -4--4- 基base )) 環丙烷Cyclopropane -1--1- 甲醯胺（Formamide ( 96b96b ）)
以與製備化合物 87a / b 類似的方法，通過SFC（ChiralPak AD-H Daicel Chemical Industries，Ltd，IC 3.0 * 25cm，5um，A：超臨界CO2 ，B：EtOH；A：B = 60/40；流量以50mL / min表示；柱溫（T）於38℃；背壓（BPR）於100bars）分離96a和96b的消旋體（19.7 mg，0.0467 mmol）），得到96a （4.00 mg，RT = 1.970 min，ee = 100.00％，20.3％產率）和96b （6.00 mg，RT = 2.803 min，ee = 99.48％，30.4％產率）。96a : LC-MS (ESI) [M+H]+ 422.1。1 H NMR (400 MHz, DMSO-d6 ): δ 11.41 (s, 1H), 8.45 (s, 1H), 7.58 (s, 1H), 7.31 - 7.26 (m, 2H ), 6.97 (s, 1H), 6.68 (s, 2H), 4.66 – 4.61 (m ,1H), 4.54 (t, J = 8.4 Hz, 1H), 4.24 – 4.21 (m, 1H), 3.77 (s, 3H), 2.29 – 2.21 (m, 2H), 1.71 – 1.66 (m, 2H), 1.43 – 1.40 (m, 1H), 1.24 – 1.20 (m, 1H), 0.84 (t, J = 8.4 Hz, 3H);96b : LC-MS (ESI) [M+H]+ 422.1。1 H NMR (400 MHz, DMSO-d6 ): δ 11.41 (s, 1H), 8.45 (s, 1H), 7.58 (s, 1H), 7.31 - 7.26 (m, 2H ), 6.97 (s, 1H), 6.68 (s, 2H), 4.66 – 4.61 (m ,1H), 4.54 (t, J = 8.4 Hz, 1H), 4.24 – 4.21 (m, 1H), 3.77 (s, 3H), 2.29 – 2.21 (m, 2H), 1.71 – 1.66 (m, 2H), 1.43 – 1.40 (m, 1H), 1.24 – 1.20 (m, 1H), 0.84 (t, J = 8.4 Hz, 3H)。In a method similar to the preparation of compound 87a/b , through SFC (ChiralPak AD-H Daicel Chemical Industries, Ltd, IC 3.0*25cm, 5um, A: supercritical CO 2 , B: EtOH; A: B = 60/40; The flow rate is expressed as 50mL/min; the column temperature (T) is at 38℃; the back pressure (BPR) is at 100bars) to separate the racemates of 96a and 96b (19.7 mg, 0.0467 mmol)) to obtain 96a (4.00 mg, R T = 1.970 min, ee = 100.00%, 20.3% yield) and 96b (6.00 mg, R T = 2.803 min, ee = 99.48%, 30.4% yield). 96a : LC-MS (ESI) [M+H] + 422.1. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.41 (s, 1H), 8.45 (s, 1H), 7.58 (s, 1H), 7.31-7.26 (m, 2H ), 6.97 (s, 1H) , 6.68 (s, 2H), 4.66 – 4.61 (m ,1H), 4.54 (t, J = 8.4 Hz, 1H), 4.24 – 4.21 (m, 1H), 3.77 (s, 3H), 2.29 – 2.21 (m , 2H), 1.71 – 1.66 (m, 2H), 1.43 – 1.40 (m, 1H), 1.24 – 1.20 (m, 1H), 0.84 (t, J = 8.4 Hz, 3H); 96b : LC-MS (ESI ) [M+H] + 422.1. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.41 (s, 1H), 8.45 (s, 1H), 7.58 (s, 1H), 7.31-7.26 (m, 2H ), 6.97 (s, 1H) , 6.68 (s, 2H), 4.66 – 4.61 (m ,1H), 4.54 (t, J = 8.4 Hz, 1H), 4.24 – 4.21 (m, 1H), 3.77 (s, 3H), 2.29 – 2.21 (m , 2H), 1.71 – 1.66 (m, 2H), 1.43 – 1.40 (m, 1H), 1.24 – 1.20 (m, 1H), 0.84 (t, J = 8.4 Hz, 3H).
( 反式 )-N-(8- 氨基 -6-((R)-4-( 氟甲基 )-2- 氧代惡唑烷 -3- 基 ) 噌啉 -3- 基 )-2-(1- 甲基 -1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺（ 97 ）的製備 
步驟1.根據製備87a / b 中描述的方法，合成為黃色固體的標題產物（150mg，37.8％產率）。LC-MS（ESI）[M + H]+ 672.3。Step 1. According to the method described in Preparation 87a/b , the title product (150 mg, 37.8% yield) was synthesized as a yellow solid. LC-MS (ESI) [M + H] + 672.3.
步驟2. 將在EtOH（2 mL）中的(反式)-N-(8-(((二苯基亞甲基)氨基)-6-((4S)-2-氧代-4-(((四氫-2H-吡喃-2-基)氧)甲基)惡唑烷-3-基)噌啉-3-基)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（100 mg，0.149 mmol）和TsOH.H2 O（3.00 mg，0.0149 mmol）的混合物在55°C下攪拌3小時。將反應混合物濃縮。通過prep-HPLC純化粗品，得到黃色固體的產物（50.0mg，79.3％產率）。LC-MS (ESI) [M+H]+ 424.0。1 H NMR (400 MHz, DMSO-d6 ): δ 11.40 (s, 1H), 8.43 (s, 1H), 7.57 (s, 1H), 7.35 (s, 1H), 7.31 (s, 1H), 6.96 (s, 1H), 6.65 (br s, 2H), 4.67 (br s, 1H), 4.51 – 4.50 (m, 1H), 4.35 – 4.34 (m, 1H), 3.77 (s, 3H), 3.69 – 3.66 (m, 1H), 3.54 – 3.51 (m, 1H), 2.28 – 2.24 (m, 2H), 1.42 – 1.41 (m, 1H), 1.22 – 1.21 (m, 1H)。Step 2. Add (trans)-N-(8-(((diphenylmethylene)amino)-6-((4S)-2-oxo-4-( ((Tetrahydro-2H-pyran-2-yl)oxy)methyl)oxazolidin-3-yl)cinnoline-3-yl)-2-(1-methyl-1H-pyrazole-4- A mixture of cyclopropane-1-carboxamide (100 mg, 0.149 mmol) and TsOH.H 2 O (3.00 mg, 0.0149 mmol) was stirred at 55°C for 3 hours. The reaction mixture was concentrated. By prep-HPLC The crude product was purified to obtain the product as a yellow solid (50.0 mg, 79.3% yield). LC-MS (ESI) [M+H] + 424.0. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.40 (s, 1H), 8.43 (s, 1H), 7.57 (s, 1H), 7.35 (s, 1H), 7.31 (s, 1H), 6.96 (s, 1H), 6.65 (br s, 2H), 4.67 (br s , 1H), 4.51 – 4.50 (m, 1H), 4.35 – 4.34 (m, 1H), 3.77 (s, 3H), 3.69 – 3.66 (m, 1H), 3.54 – 3.51 (m, 1H), 2.28 – 2.24 (m, 2H), 1.42 – 1.41 (m, 1H), 1.22 – 1.21 (m, 1H).
步驟3.在0°C條件下，在(反式)-N-(8-氨基-6- ((S)-4-(羥甲基)-2-氧代惡唑烷-3-基)噌啉-3-基)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（40.0 mg，0.0946 mmol）和Et3 N（86.0 mg，0.852 mmol）的乙腈（2 mL）溶液中緩慢加入PBSF（全氟-1-丁基磺醯氟）（86.0 mg，0.284 mmol）。將所得混合物在0℃下攪拌5min。然後加入Et3 N.3HF（46.0mg，0.284mmol）。將反應混合物在0℃下攪拌1小時。將混合物用NaHCO3 水溶液（10 mL）稀釋，並用乙酸乙酯（10 mL * 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓下濃縮。通過prep-HPLC純化粗品，得到呈黃色固體的產物（4.21mg，10.5％產率）。LC-MS (ESI) [M+H]+ 426.0。1 H NMR (400 MHz, DMSO-d6 ): δ 11.44 (s, 1H), 8.46 (s, 1H), 7.58 (s, 1H), 7.36 (s, 1H), 7.31 (s, 1H), 6.96 (s, 1H), 6.72 (s, 2H), 5.00 – 4.93 (m, 1H), 4.78 – 4.53 (m, 3H), 4.41 – 4.39 (m, 1H), 3.77 (s, 3H), 2.41 – 2.22 (m, 2H), 1.42 – 1.41 (m, 1H), 1.22 – 1.21 (m, 1H)。Step 3. At 0°C, in (trans)-N-(8-amino-6- ((S)-4-(hydroxymethyl)-2-oxooxazolidine-3-yl) Cinolin-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-carboxamide (40.0 mg, 0.0946 mmol) and Et 3 N (86.0 mg, 0.852 mmol) Slowly add PBSF (perfluoro-1-butanesulfon fluoride) (86.0 mg, 0.284 mmol) to the acetonitrile (2 mL) solution. The resulting mixture was stirred at 0°C for 5 min. Then Et 3 N.3HF (46.0 mg, 0.284 mmol) was added. The reaction mixture was stirred at 0°C for 1 hour. The mixture was diluted with aqueous NaHCO 3 (10 mL) and extracted with ethyl acetate (10 mL * 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by prep-HPLC to obtain the product as a yellow solid (4.21 mg, 10.5% yield). LC-MS (ESI) [M+H] + 426.0. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.44 (s, 1H), 8.46 (s, 1H), 7.58 (s, 1H), 7.36 (s, 1H), 7.31 (s, 1H), 6.96 (s, 1H), 6.72 (s, 2H), 5.00 – 4.93 (m, 1H), 4.78 – 4.53 (m, 3H), 4.41 – 4.39 (m, 1H), 3.77 (s, 3H), 2.41 – 2.22 (m, 2H), 1.42 – 1.41 (m, 1H), 1.22 – 1.21 (m, 1H).
（反式） -N- （ 8- 氨基 -6- （（ S ） -4- （異丙基） -2- 氧代惡唑烷 -3- 基）噌啉 -3- 基） -2- （ 1- 甲基 -1H- 吡唑 -4- 基）環丙烷 -1- 甲醯胺（ 98 ）的製備 
步驟1.根據製備87a / b 中描述的方法，合成為黃色油的(反式)-N-(8-(((二苯基亞甲基)氨基)-6-((S)-4-異丙基-2-氧代惡唑烷-3-基)噌啉-3-基)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（28.0mg，29.6％產率）。LC-MS（ESI）[M + H]+ 600.4。Step 1. According to the method described in preparation 87a/b , synthesize (trans)-N-(8-(((diphenylmethylene)amino)-6-((S)-4- Isopropyl-2-oxooxazolidin-3-yl) cinnoline-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-methanamide ( 28.0 mg, 29.6% yield) LC-MS (ESI) [M + H] + 600.4.
步驟2.根據通用步驟C ，合成為黃色固體的(反式)-N-(8-氨基-6-((S)-4-異丙基-2-氧代惡唑烷-3-基)噌啉-3-基)-2-(1 -甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（98 ）（8.93mg，34.3％產率）。LC-MS (ESI) [M+H]+ 436.4。1 H NMR (400 MHz, DMSO-d6): δ 11.42 (s, 1H), 8.46 (d, J = 1.6 Hz, 1H), 7.58 (s, 1H), 7.31 (s, 1H), 7.26 (t, J = 2.0 Hz, 1H ), 7.01 (s, 1H), 6.69 (s, 2H), 4.68 – 4.65 (m, 1H), 4.45 (t, J = 8.8 Hz, 1H), 4.35 – 4.32 (m, 1H), 3.78 (s, 3H), 2.29 – 2.16 (m, 3H), 1.43 – 1.39 (m, 1H), 1.25 – 1.23 (m, 1H), 0.90 (d, J = 7.2 Hz, 3H), 0.76 (d, J = 6.8 Hz, 3H)。Step 2. According to General Procedure C , (trans)-N-(8-amino-6-((S)-4-isopropyl-2-oxooxazolidin-3-yl) was synthesized as a yellow solid Cinolin-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-carboxamide ( 98 ) (8.93 mg, 34.3% yield). LC-MS (ESI) [M+H]+ 436.4. 1 H NMR (400 MHz, DMSO-d6): δ 11.42 (s, 1H), 8.46 (d, J = 1.6 Hz, 1H), 7.58 (s, 1H), 7.31 (s, 1H), 7.26 (t, J = 2.0 Hz, 1H ), 7.01 (s, 1H), 6.69 (s, 2H), 4.68 – 4.65 (m, 1H), 4.45 (t, J = 8.8 Hz, 1H), 4.35 – 4.32 (m, 1H ), 3.78 (s, 3H), 2.29 – 2.16 (m, 3H), 1.43 – 1.39 (m, 1H), 1.25 – 1.23 (m, 1H), 0.90 (d, J = 7.2 Hz, 3H), 0.76 ( d, J = 6.8 Hz, 3H).
( 反式 )-N-(8- 氨基 -6-(8- 甲基 -2,3- 二氫 -1H- 吡啶並 [2,3-b] [1,4] 惡嗪 -7- 基 ) 噌啉 -3 - 基） -2-(1-(3-( 甲基磺醯基 ) 丙基 )-1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺（ 99 ）的製備 
步驟1. 在室溫下，向（反式）-2-（1H-吡唑-4-基）環丙烷-1-甲酸乙酯（62.0 mg，0.344 mmol）和碳酸銫（223 mg，0.684 mmol）在N，N-二甲基甲醯胺（2.00 mL）中的懸浮液中加入4-甲基苯磺酸3-（甲基磺醯基）丙基酯（100 mg，0.342 mmol）。將混合物在100℃下攪拌18小時。將混合物冷卻至室溫，然後用水（20.0mL）稀釋，用乙酸乙酯（20.0mL * 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓下濃縮，得到粗產物。殘餘物通過矽膠柱色譜法（石油醚/乙酸乙酯= 10/1）純化，得到為白色固體的（反式）-2-（1-（3-（甲基磺醯基）丙基）-1H-吡唑-4-基）環丙烷-1-羧酸乙基酯（100mg，產率97.3％）。LC-MS（ESI）[M + H]+ 301.0。Step 1. At room temperature, add (trans)-2-(1H-pyrazol-4-yl)cyclopropane-1-ethyl carboxylate (62.0 mg, 0.344 mmol) and cesium carbonate (223 mg, 0.684 mmol) ) Add 3-(methylsulfonyl)propyl 4-methylbenzenesulfonate (100 mg, 0.342 mmol) to the suspension in N,N-dimethylformamide (2.00 mL). The mixture was stirred at 100°C for 18 hours. The mixture was cooled to room temperature, then diluted with water (20.0 mL), and extracted with ethyl acetate (20.0 mL * 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1) to obtain (trans)-2-(1-(3-(methylsulfonyl)propyl)- as a white solid 1H-pyrazol-4-yl)cyclopropane-1-carboxylic acid ethyl ester (100 mg, yield 97.3%). LC-MS (ESI) [M + H] + 301.0.
步驟2-4。根據製備化合物73a / 73b 中描述的方法，合成為黃色固體化合物 99 （34.9 mg，產率64.3％）。LC-MS (ESI) [M+H]+ 563.1。1 H NMR (400 MHz, DMSO-d6 ): δ 11.48 (s, 1H), 8.51 (s, 1H), 7.67 (s, 1H), 7.38 (s, 1H), 7.35 (s, 1H), 6.84 (s, 1H), 6.68 (d, J = 1.2 Hz, 1H), 6.63 (s, 2H), 5.67 (s, 1H), 4.28 - 4.27 (m, 2H), 4.17 (t, J = 6.8 Hz, 2H), 3.37 - 3.36 (m, 2H), 3.10 - 3.06 (m, 2H), 2.99 (s, 3H), 2.33 - 2.24 (m, 2H), 2.19 - 2.15 (m, 2H), 2.03 (s, 3H), 1.45 - 1.42 (m, 1H), 1.26 - 1.23 (m, 1H)。Steps 2-4. According to the method described in the preparation of compound 73a/73b , compound 99 (34.9 mg, yield 64.3%) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 563.1. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.48 (s, 1H), 8.51 (s, 1H), 7.67 (s, 1H), 7.38 (s, 1H), 7.35 (s, 1H), 6.84 (s, 1H), 6.68 (d, J = 1.2 Hz, 1H), 6.63 (s, 2H), 5.67 (s, 1H), 4.28-4.27 (m, 2H), 4.17 (t, J = 6.8 Hz, 2H), 3.37-3.36 (m, 2H), 3.10-3.06 (m, 2H), 2.99 (s, 3H), 2.33-2.24 (m, 2H), 2.19-2.15 (m, 2H), 2.03 (s, 3H), 1.45-1.42 (m, 1H), 1.26-1.23 (m, 1H).
根據與化合物According to the compound 9999 類似的方法製備化合物Similar methods to prepare compounds 100-103100-103 。.
（( 反式Trans ）) -N--N- （( 8-8- 氨基Amino -6--6- （( 8-8- 甲基methyl -2,3--2,3- 二氫Dihydro -1H--1H- 吡啶並Pyrido [2,3-b] [1,4][2,3-b] [1,4] 惡嗪Oxazine -7--7- 基base ）) 噌啉Cinolin -3--3- 基base ）) -2--2- （( 1-1- （( 2-2- 羥乙基Hydroxyethyl ）) -1H--1H- 吡唑Pyrazole -4--4- 基base ）) 環丙烷Cyclopropane -1--1- 甲醯胺Formamide （( 100100 ）)
LC-MS (ESI) [M+H]+ 487.2.1 H NMR (400 MHz, DMSO-d6 ): δ 11.49 (s, 1H), 8.51 (s, 1H), 7.61 (s, 1H), 7.33 (d, J = 6.0 Hz, 2H), 6.84 (s, 1H), 6.67 – 6.62 (m, 3H), 5.67 (s, 1H), 4.89 – 4.86 (m, 1H), 4.86 (s, 2H), 4.07 (t, J = 4.8 Hz, 2H), 3.70 (t, J = 4.8 Hz, 2H), 3.39 – 3.35 (m, 2H), 2.32 – 2.29 (m, 2H), 2.03 (s, 3H), 1.43 – 1.40 (m, 1H), 1.24 – 1.23 (m, 1H).LC-MS (ESI) [M+H] + 487.2. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.49 (s, 1H), 8.51 (s, 1H), 7.61 (s, 1H), 7.33 (d, J = 6.0 Hz, 2H), 6.84 (s, 1H), 6.67 – 6.62 (m, 3H), 5.67 (s, 1H), 4.89 – 4.86 (m, 1H), 4.86 (s, 2H), 4.07 (t, J = 4.8 Hz, 2H), 3.70 (t, J = 4.8 Hz, 2H), 3.39 – 3.35 (m, 2H), 2.32 – 2.29 (m, 2H), 2.03 (s, 3H), 1.43 – 1.40 (m, 1H), 1.24 – 1.23 (m, 1H).
（反式）(Trans) -N--N- （( 8-8- 氨基Amino -6--6- （( 8-8- 甲基methyl -2,3--2,3- 二氫Dihydro -1H--1H- 吡啶並Pyrido [2,3-b] [1,4][2,3-b] [1,4] 惡嗪Oxazine -7--7- 基）噌啉Base) Cinolin -3--3- 基base ）) -2--2- （( 1-1- （( 2-2- （甲基磺醯基）乙基）(Methylsulfonyl) ethyl) -1H--1H- 吡唑Pyrazole -4--4- 基）環丙烷Base) cyclopropane -1--1- 甲醯胺（Formamide ( 101101 ）)
LC-MS (ESI) [M+H]+ 549.6.1 H NMR (400 MHz, DMSO-d6 ) δ 11.50 (s, 1H), 8.51 (s, 1H), 7.73 (s, 1H), 7.42 (s, 1H), 7.35 (s, 1H), 6.85 (s, 1H), 6.68 (s, 1H), 6.63 (s, 2H), 5.67 (s, 1H), 4.50 – 4.46 (m, 2H), 4.27 (s, 2H), 3.69 – 3.65 (m, 2H), 3.36 (s, 2H), 2.85 (s, 3H), 2.32 – 2.26 (m, 2H), 2.03 (s, 3H), 1.49 – 1.42 (m, 1H), 1.27 – 1.23 (m, 1H)。LC-MS (ESI) [M+H] + 549.6. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.50 (s, 1H), 8.51 (s, 1H), 7.73 (s, 1H), 7.42 ( s, 1H), 7.35 (s, 1H), 6.85 (s, 1H), 6.68 (s, 1H), 6.63 (s, 2H), 5.67 (s, 1H), 4.50 – 4.46 (m, 2H), 4.27 (s, 2H), 3.69 – 3.65 (m, 2H), 3.36 (s, 2H), 2.85 (s, 3H), 2.32 – 2.26 (m, 2H), 2.03 (s, 3H), 1.49 – 1.42 (m , 1H), 1.27 – 1.23 (m, 1H).
（反式）(Trans) -2--2- （( 1-1- （( 1-1- （( 1-1- 乙醯氮雜環丁烷Acetidine -3--3- 基）base) -1H--1H- 吡唑Pyrazole -4--4- 基）base) -N--N- （( 8-8- 氨基Amino -6--6- （( 8-8- 甲基methyl -2,3--2,3- 二氫Dihydro - 1H--1H- 吡啶並Pyrido [2,3-b] [1,4][2,3-b] [1,4] 惡嗪Oxazine -7--7- 基）噌啉Base) Cinolin -3--3- 基）環丙烷Base) cyclopropane -1--1- 甲醯胺（Formamide ( 102102 ）的製備) Preparation
LC-MS (ESI) [M+H]+ 540.6.1 H NMR (400 MHz, DMSO-d6 ) δ 11.48 (s, 1H), 8.51 (s, 1H), 7.83 (s, 1H), 7.49 (s, 1H), 7.35 (s, 1H), 6.84 (d, J =1.6 Hz, 1H), 6.68 (d, J =1.6 Hz, 1H), 6.63 (s, 2H), 5.67 (s, 1H), 5.19 – 5.15 (m, 1H), 4.56 – 4.52 (m, 1H), 4.37 – 4.33 (m, 1H), 4.28 – 4.24 (m, 3H), 4.07 – 4.04 (m, 1H), 3.37 (s, 2H), 2.33 – 2.24 (m, 2H), 2.03 (s, 3 H), 1.80 (s, 3 H), 1.46 – 1.43 (m, 1H), 1.30 – 1.27 (m, 1H)。LC-MS (ESI) [M+H] + 540.6. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.48 (s, 1H), 8.51 (s, 1H), 7.83 (s, 1H), 7.49 ( s, 1H), 7.35 (s, 1H), 6.84 (d, J =1.6 Hz, 1H), 6.68 (d, J =1.6 Hz, 1H), 6.63 (s, 2H), 5.67 (s, 1H), 5.19 – 5.15 (m, 1H), 4.56 – 4.52 (m, 1H), 4.37 – 4.33 (m, 1H), 4.28 – 4.24 (m, 3H), 4.07 – 4.04 (m, 1H), 3.37 (s, 2H) ), 2.33 – 2.24 (m, 2H), 2.03 (s, 3 H), 1.80 (s, 3 H), 1.46 – 1.43 (m, 1H), 1.30 – 1.27 (m, 1H).
（反式）(Trans) -N--N- （( 8-8- 氨基Amino -6--6- （( 8-8- 甲基methyl -2,3--2,3- 二氫Dihydro -1H--1H- 吡啶並Pyrido [2,3-b] [1,4][2,3-b] [1,4] 惡嗪Oxazine -7--7- 基）噌啉Base) Cinolin -3 --3- 基）base) -2--2- （( 1-1- （( 1-1- （( 1-1- （甲基磺醯基）氮雜環丁烷(Methylsulfonyl)azetidine -3--3- 基）base) -1H--1H- 吡唑Pyrazole -4--4- 基）環丙烷Base) cyclopropane -1--1- 甲醯胺（Formamide ( 103103 ）的製備) Preparation
LC-MS (ESI) [M+H]+ 576.6.1 H NMR (400 MHz, DMSO-d6 ) δ 11.49 (s, 1H), 8.51 (s, 1H), 7.81 (s, 1H), 7.52 (s, 1H), 7.35 (s, 1H), 6.84 (d, J =1.2 Hz, 1H), 6.68 (d, J =1.2 Hz, 1H), 6.63 (s, 2H), 5.67 (s, 1H), 5.23 – 5.20 (m, 1H), 4.28 – 4.20 (m, 6H), 3.36 – 3.35 (m, 2H), 3.12 (s, 3 H), 2.33 – 2.26 (m, 2H), 2.03 (s, 3 H), 1.45 – 1.43 (m, 1H), 1.29 – 1.27 (m, 1H)。LC-MS (ESI) [M+H] + 576.6. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.49 (s, 1H), 8.51 (s, 1H), 7.81 (s, 1H), 7.52 ( s, 1H), 7.35 (s, 1H), 6.84 (d, J =1.2 Hz, 1H), 6.68 (d, J =1.2 Hz, 1H), 6.63 (s, 2H), 5.67 (s, 1H), 5.23 – 5.20 (m, 1H), 4.28 – 4.20 (m, 6H), 3.36 – 3.35 (m, 2H), 3.12 (s, 3 H), 2.33 – 2.26 (m, 2H), 2.03 (s, 3 H) ), 1.45 – 1.43 (m, 1H), 1.29 – 1.27 (m, 1H).
( 反式 )-N-(8- 氨基 -6-(8- 甲基 -2,3- 二氫 -1H- 吡啶並 [2,3-b] [1,4] 惡嗪 -7- 基 ) 噌啉 -3 - 基 )-2-(4-( 甲基磺醯基 )-1H- 吡唑 -1- 基 ) 環丙烷 -1- 甲醯胺（ 104 ）的製備 
步驟1. 在室溫下，向1-（2-溴乙基）-4-碘-1H-吡唑（2.00 g，6.64 mmol）在DMSO（10.0 mL）中的溶液中加入DBU（1,8-二氮雜雙環 [5.4.0] 十一碳-7-烯）（5.00 mL）。將混合物在80℃下攪拌18小時。將混合物冷卻至室溫。用水（50.0mL）稀釋混合物，並用乙酸乙酯（20.0mL * 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓下濃縮。將殘餘物通過矽膠柱色譜法（石油醚/乙酸乙酯= 5/1）純化，得到呈無色油狀的4-碘-1-乙烯基-1H-吡唑（1.20g，51.5％產率）。LC-MS（ESI）[M + H]+ 220.7。Step 1. To a solution of 1-(2-bromoethyl)-4-iodo-1H-pyrazole (2.00 g, 6.64 mmol) in DMSO (10.0 mL) at room temperature, add DBU (1,8 -Diazabicyclo[5.4.0] undec-7-ene) (5.00 mL). The mixture was stirred at 80°C for 18 hours. The mixture was cooled to room temperature. The mixture was diluted with water (50.0 mL) and extracted with ethyl acetate (20.0 mL * 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1) to obtain 4-iodo-1-vinyl-1H-pyrazole as a colorless oil (1.20 g, 51.5% yield) . LC-MS (ESI) [M + H] + 220.7.
步驟2. 在0℃下，向4-碘-1-乙烯基-1H-吡唑（1.70g，7.73mmol）在二氯甲烷（20.0mL）中的溶液中加入乙酸銠（II）二聚體（500mg）和2-重氮乙酸乙酯（1.76 g，15.5mmol）。將混合物在室溫攪拌18小時，然後用水（30.0mL）稀釋，用二氯甲烷（20.0mL * 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓下濃縮。殘餘物通過矽膠柱色譜法（石油醚/乙酸乙酯= 3/1）純化，得到為棕色固體的（反式）-2-（4-碘-1H-吡唑-1-基）環丙烷-1-羧酸乙酯（220mg）（220mg，9.32％產率）。LC-MS（ESI）[M + H]+ 306.9。Step 2. Add rhodium(II) acetate dimer to a solution of 4-iodo-1-vinyl-1H-pyrazole (1.70g, 7.73mmol) in dichloromethane (20.0mL) at 0°C (500 mg) and ethyl 2-diazoacetate (1.76 g, 15.5 mmol). The mixture was stirred at room temperature for 18 hours, then diluted with water (30.0 mL) and extracted with dichloromethane (20.0 mL * 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 3/1) to obtain (trans)-2-(4-iodo-1H-pyrazol-1-yl)cyclopropane- as a brown solid Ethyl 1-carboxylate (220 mg) (220 mg, 9.32% yield). LC-MS (ESI) [M + H] + 306.9.
步驟3. 在室溫下，向（反式）-2-（4-碘-1H-吡唑-1-基）環丙烷-1-甲酸乙酯（170 mg，0.556 mmol）和甲烷亞磺酸鈉（114 mg，1.12 mmol）的DMSO（4.00mL）溶液中加入（反式）-N 1，N 2-二甲基環己烷-1，2-二胺（16mg，0.112mmol）和Cu(OTf)2 （40mg，0.112mmol）。將混合物在110℃下攪拌18小時。將混合物冷卻至室溫，並用水（30.0mL）稀釋，用乙酸乙酯（20.0mL * 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓下濃縮。殘餘物通過矽膠柱色譜法（石油醚/乙酸乙酯= 2/1）純化，得到呈棕色固體的（反式）-2-（4-（甲基磺醯基）-1H-吡唑-1-基）環丙烷-1-羧酸乙酯（ 75.0mg，52.4％產率）。LC-MS（ESI）[M + H]+ 258.9。Step 3. At room temperature, add ethyl (trans)-2-(4-iodo-1H-pyrazol-1-yl)cyclopropane-1-carboxylate (170 mg, 0.556 mmol) and methanesulfinic acid To a solution of sodium (114 mg, 1.12 mmol) in DMSO (4.00 mL) was added (trans)-N 1,N 2-dimethylcyclohexane-1,2-diamine (16 mg, 0.112 mmol) and Cu( OTf) 2 (40mg, 0.112mmol). The mixture was stirred at 110°C for 18 hours. The mixture was cooled to room temperature, diluted with water (30.0 mL), and extracted with ethyl acetate (20.0 mL * 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 2/1) to obtain (trans)-2-(4-(methylsulfonyl)-1H-pyrazole-1 as a brown solid) -Yl) ethyl cyclopropane-1-carboxylate (75.0 mg, 52.4% yield). LC-MS (ESI) [M + H] + 258.9.
步驟4-6.根據製備化合物 73a / 73b 描述的方法，合成了一種呈黃色固體的化合物 104 （27.0mg，50.9％產率）。LC-MS (ESI) [M+H]+ 521.1。1 H NMR (400 MHz, DMSO-d6 ): δ 8.69 (s, 1H), 8.52 (s, 1H), 7.98 (s, 1H), 7.35 (s, 1H), 6.87 (d, J = 1.6 Hz, 1H), 6.70 (d, J = 1.6 Hz, 1H), 6.66 (s, 2H), 6.06 (s, 1H), 5.68 (s, 1H), 4.31 - 4.26 (m, 3H), 3.36 - 3.32 (m, 2H), 3.22 (s, 3H), 2.90 - 2.85 (m, 1H), 2.03 (s, 3H), 1. 94 - 1.89 (m, 1H), 1. 67 - 1.62 (m, 1H)。Steps 4-6. According to the method described for preparing compound 73a/73b , compound 104 (27.0 mg, 50.9% yield) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 521.1. 1 H NMR (400 MHz, DMSO-d 6 ): δ 8.69 (s, 1H), 8.52 (s, 1H), 7.98 (s, 1H), 7.35 (s, 1H), 6.87 (d, J = 1.6 Hz , 1H), 6.70 (d, J = 1.6 Hz, 1H), 6.66 (s, 2H), 6.06 (s, 1H), 5.68 (s, 1H), 4.31-4.26 (m, 3H), 3.36-3.32 ( m, 2H), 3.22 (s, 3H), 2.90-2.85 (m, 1H), 2.03 (s, 3H), 1.94-1.89 (m, 1H), 1. 67-1.62 (m, 1H).
2-(6-((8- 氨基 -6-(4- 甲基吡啶 -3- 基 ) 噌啉 3- 基 ) 氨基 ) 吡啶 -3- 基 ) 乙腈 （ 105 ） 的製備 
步驟1. 在室溫下，在8-((二苯基亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-胺（50mg，0.120mmol）和2-(6-氯吡啶-3-基)乙腈（37 mg，0.295 mmol）的二氧雜環己烷（2.00 mL）溶液中加入4,5-雙(二苯基膦基)-9,9-二甲基氧雜蒽（28 mg，0.0.0482 mmol）、碳酸銫（118 mg，0.362 mmol）和三(二亞苄基丙酮)二鈀（22 mg，0.0241 mmol）。將該混合物在Ar下於150℃的微波中照射2小時。冷卻至室溫後，將混合物用水（30mL）稀釋，並用乙酸乙酯（20.0mL * 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓下濃縮。殘餘物通過柱色譜法（二氯甲烷/甲醇= 20/1）純化，得到為黃色固體的2-(6-((8-((二苯基亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-基)氨基)吡啶-3-基)乙腈（30mg，產率46.9％）。LC-MS（ESI）[M + H]+ 532.0。Step 1. At room temperature, add 8-((diphenylmethylene)amino)-6-(4-methylpyridin-3-yl)cinnoline-3-amine (50mg, 0.120mmol) and 2 -(6-Chloropyridin-3-yl)acetonitrile (37 mg, 0.295 mmol) in dioxane (2.00 mL) was added 4,5-bis(diphenylphosphino)-9,9- Dimethylxanthene (28 mg, 0.0.0482 mmol), cesium carbonate (118 mg, 0.362 mmol) and tris(dibenzylideneacetone) dipalladium (22 mg, 0.0241 mmol). The mixture was irradiated in a microwave at 150°C under Ar for 2 hours. After cooling to room temperature, the mixture was diluted with water (30 mL) and extracted with ethyl acetate (20.0 mL * 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane/methanol = 20/1) to obtain 2-(6-((8-((diphenylmethylene)amino)-6-(4- Methylpyridin-3-yl) cinnolin-3-yl)amino)pyridin-3-yl)acetonitrile (30 mg, yield 46.9%). LC-MS (ESI) [M + H] + 532.0.
步驟2.根據通用步驟C ，合成為黃色固體的化合物 105 （11.0mg， 64.7％產率）。LC-MS (ESI) [M+H]+ 368.0。1 H NMR (400 MHz, DMSO-d6 ): δ 10.48 (s, 1H), 8.61 (s, 1H), 8.47 - 8.45 (m, 2H), 8.25 (s, 1H), 7.72 - 7.69 (m, 1H), 7.41 - 7.36 (m, 2H), 6.90 (s, 1H), 6.63 - 6.61 (m, 3H), 3.99 (s, 2H), 2.33 (s, 3H)。Step 2. According to general procedure C , compound 105 (11.0 mg, 64.7% yield) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 368.0. 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.48 (s, 1H), 8.61 (s, 1H), 8.47-8.45 (m, 2H), 8.25 (s, 1H), 7.72-7.69 (m, 1H), 7.41-7.36 (m, 2H), 6.90 (s, 1H), 6.63-6.61 (m, 3H), 3.99 (s, 2H), 2.33 (s, 3H).
(6-((8- 氨基 -6-(4- 甲基吡啶 -3- 基 ) 噌啉 -3- 基 ) 氨基 ) 吡啶 -3- 基 )( 嗎啉代 ) 甲酮 (106) 的製備 
步驟1.在8-((二苯基亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-胺（250 mg，0.602 mmol）和6-溴煙酸甲酯（110 mg，0.509 mmol）的二氧雜環己烷（15 mL）溶液中加入Pd(OAc)2 （12.5 mg，0.0509 mmol）、CyJhonPhos（35 mg，0.101 mmol）和Cs2 CO3 （490 mg，1.527 mmol）。在氬氣氣氛下於100°C攪拌16小時後，將反應混合物冷卻至室溫，用水（100 mL）稀釋，並用DCM（50 mL * 3）萃取。合併的有機層經Na2 SO4 乾燥並真空濃縮，得到殘餘物，將其通過prep-TLC（PE：EA = 2：1，v / v）純化，得到為黃色固體的標題化合物（100mg，收率35.7％）。LC-MS（ESI）[M + H]+ 551.2。Step 1. In 8-((diphenylmethylene)amino)-6-(4-methylpyridin-3-yl)cinolin-3-amine (250 mg, 0.602 mmol) and 6-bromonicotinic acid Pd(OAc) 2 (12.5 mg, 0.0509 mmol), CyJhonPhos (35 mg, 0.101 mmol) and Cs 2 CO 3 ( 490 mg, 1.527 mmol). After stirring at 100°C for 16 hours under an argon atmosphere, the reaction mixture was cooled to room temperature, diluted with water (100 mL), and extracted with DCM (50 mL * 3). The combined organic layer was dried over Na 2 SO 4 and concentrated in vacuo to obtain a residue, which was purified by prep-TLC (PE:EA = 2:1, v/v) to obtain the title compound (100 mg, yield) as a yellow solid. Rate 35.7%). LC-MS (ESI) [M + H] + 551.2.
步驟2.向6-((8-((二苯基亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-基)氨基)煙酸甲酯（100 mg，0.182 mmol）的DMSO/ H2 O（4.00mL / 1mL）溶液中加入LiOH·H2 O（76.0mg，1.82mmol）。在30°C攪拌16小時後，將混合物用H2 O（20 mL）稀釋，用HCl（1N）酸化至pH = 5，並用乙酸乙酯（15 mL * 3）萃取。合併的有機層用鹽水（15mL×3）洗滌，經Na2 SO4 乾燥，過濾並真空濃縮得到殘餘物，將其通過prep-TLC純化（DCM：MeOH = 20：1，v / v），得到為白色固體的標題化合物（80.0 mg，產率82.0％）。LC-MS（ESI）[M + H]+ 537.0。Step 2. To 6-((8-((diphenylmethylene)amino)-6-(4-methylpyridin-3-yl)cinnolin-3-yl)amino)nicotinic acid methyl ester (100 mg, 0.182 mmol) in DMSO/H 2 O (4.00 mL / 1 mL) solution was added LiOH·H 2 O (76.0 mg, 1.82 mmol). After stirring for 16 hours at 30°C, the mixture was diluted with H 2 O (20 mL), acidified with HCl (1N) to pH = 5, and extracted with ethyl acetate (15 mL * 3). The combined organic layer was washed with brine (15mL×3), dried over Na 2 SO 4 , filtered and concentrated in vacuo to obtain a residue, which was purified by prep-TLC (DCM:MeOH = 20:1, v/v) to obtain The title compound (80.0 mg, yield 82.0%) as a white solid. LC-MS (ESI) [M + H] + 537.0.
步驟3.在6-(((8-((二苯基亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-基)氨基)煙酸（40.0 mg，0.074 mmol）和嗎啉（1.5eq）在DCM（2mL）的溶液中加入TEA（22.5mg，0.223mmol）和HATU（36mg，0.962mmol）。在室溫下攪拌16小時後，將混合物用H2 O（20mL）稀釋並用EA（10mL * 3）萃取。合併的有機相經Na2 SO4 乾燥，並真空濃縮，得到殘餘物，將其通過prep-TLC純化（DCM：MeOH = 20：1，v / v），得到為黃色固體的標題化合物（20mg，收率48.0％）。LC-MS（ESI）[M + H]+ 606.2。Step 3. In 6-(((8-((diphenylmethylene)amino)-6-(4-methylpyridin-3-yl)cinolin-3-yl)amino)nicotinic acid (40.0 mg , 0.074 mmol) and morpholine (1.5eq) in DCM (2mL) were added TEA (22.5mg, 0.223mmol) and HATU (36mg, 0.962mmol). After stirring at room temperature for 16 hours, the mixture was washed with H 2 O (20 mL) was diluted and extracted with EA (10 mL * 3). The combined organic phase was dried over Na 2 SO 4 and concentrated in vacuo to obtain a residue, which was purified by prep-TLC (DCM:MeOH = 20:1, v / v), the title compound (20 mg, yield 48.0%) was obtained as a yellow solid. LC-MS (ESI) [M + H] + 606.2.
步驟4.根據通用步驟C 中描述的方法，合成為黃色固體的化合物 106 （1.01mg，產率6.94％）。LC-MS (ESI) [M+H]+ 442.1.1 H NMR (400 MHz, DMSO-d6 ): δ 8.79 (s, 1H), 8.43 – 8.40 (m, 3H), 7.81 – 7.74 (m, 1H), 7.40 (d, J = 5.2 Hz, 1H), 7.30 (d, J = 8.8 Hz, 1H), 6.95 (d, J = 1.6 Hz, 1H), 6.73 (d, J = 1.2 Hz, 1H), 3.85 – 3.70 (m ,8H), 2.40 (s, 3H)。Step 4. According to the method described in General Step C , compound 106 (1.01 mg, yield 6.94%) was synthesized as a yellow solid. LC-MS (ESI) [M+H] + 442.1. 1 H NMR (400 MHz, DMSO-d 6 ): δ 8.79 (s, 1H), 8.43 – 8.40 (m, 3H), 7.81 – 7.74 (m, 1H), 7.40 (d, J = 5.2 Hz, 1H), 7.30 (d, J = 8.8 Hz, 1H), 6.95 (d, J = 1.6 Hz, 1H), 6.73 (d, J = 1.2 Hz, 1H) , 3.85 – 3.70 (m ,8H), 2.40 (s, 3H).
( 反式 )-N- ( 8- 氨基 -7- 氯 -6- (( S)-4- 甲基 -2- 氧代惡唑烷 -3- 基 ) 噌啉 -3- 基 )-2- ( 1- 甲基 - 1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺（ 107 ）的製備 
在室溫下，向(反式)-N-(8-氨基-6-((S)-4-甲基-2-氧代惡唑烷-3-基)噌啉-3-基)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-羧醯胺（50mg，0.123mmol）的乙腈（5.00mL）溶液中加入N-氯琥珀醯亞胺（13mg，0.0982mmol）。將混合物在50℃下攪拌3小時。冷卻至室溫後，將混合物通過Pre-HPLC法（NH3 . H2 O）純化，得到呈黃色固體的(反式)-N-(8-氨基-7-氯-6-((S)-4-甲基-2-)氧代惡唑烷-3-基)噌啉-3-基)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（27.83 mg，51.5％產率）。LC-MS (ESI) [M+H]+ 442.0。1 H NMR (400 MHz, DMSO-d6 ): δ 11.73 (s, 1H), 8.82 (s, 1H), 7.59 (s, 1H), 7.32 (s, 1H), 7.00 (s, 2H), 6.76 (s, 1H), 4.72 - 4.67 (m, 1H), 4.58 - 4.53 (m, 1H), 4.11 - 4.07 (m, 1H), 3.78 (s, 3H), 2.33 - 2.26 (m, 2H), 1.48 - 1.44 (m, 1H), 1.29 - 1.24 (m, 1H), 1.15 (d, J = 6.0 Hz, 3H)。At room temperature, to (trans)-N-(8-amino-6-((S)-4-methyl-2-oxooxazolidin-3-yl) cinnolin-3-yl)- 2-(1-Methyl-1H-pyrazol-4-yl)cyclopropane-1-carboxamide (50mg, 0.123mmol) in acetonitrile (5.00mL) was added with N-chlorosuccinimide (13mg, 0.0982mmol). The mixture was stirred at 50°C for 3 hours. After cooling to room temperature, the mixture was purified by Pre-HPLC method (. NH 3 H 2 O) to give a yellow solid (trans) -N- (8- amino-7-chloro -6 - ((S) -4-Methyl-2-)oxazolidin-3-yl)cinolin-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-methyl Amide (27.83 mg, 51.5% yield). LC-MS (ESI) [M+H] + 442.0. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.73 (s, 1H), 8.82 (s, 1H), 7.59 (s, 1H), 7.32 (s, 1H), 7.00 (s, 2H), 6.76 (s, 1H), 4.72-4.67 (m, 1H), 4.58-4.53 (m, 1H), 4.11-4.07 (m, 1H), 3.78 (s, 3H), 2.33-2.26 (m, 2H), 1.48 -1.44 (m, 1H), 1.29-1.24 (m, 1H), 1.15 (d, J = 6.0 Hz, 3H).
( 反式 )-N-(8- 氨基 -5- 氯 -6-((S)-4- 甲基 -2- 氧代惡唑烷 -3- 基 ) 噌啉 -3- 基 )-2-(1- 甲基 - 1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺（ 108 ）的製備 
步驟1. 在0℃下，向(反式)-N-(8-氨基-6-((S)-4-甲基-2-氧代惡唑烷-3-基)噌啉-3-基)）-2-(1-甲基-1 H-吡唑-4-基)環丙烷-1-甲醯胺（100mg，0.245mmol）在DMF（3mL）的溶液中緩慢加入NIS（66.0 mg, 0.295 mmol）。在室溫下攪拌3小時後，將混合物用水（10mL）稀釋，並用乙酸乙酯（10mL * 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓下濃縮。殘餘物通過prep-TLC（DCM / MeOH = 20/1）純化，得到呈棕色固體的產物（130mg，99.5％產率）。LC-MS（ESI）[M + H]+ 534.3。Step 1. At 0℃, add (trans)-N-(8-amino-6-((S)-4-methyl-2-oxooxazolidin-3-yl) cinnoline-3- Yl))-2-(1-methyl-1 H-pyrazol-4-yl)cyclopropane-1-carboxamide (100mg, 0.245mmol) in DMF (3mL) solution was slowly added NIS (66.0 mg , 0.295 mmol). After stirring at room temperature for 3 hours, the mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL * 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by prep-TLC (DCM / MeOH = 20/1) to obtain the product (130 mg, 99.5% yield) as a brown solid. LC-MS (ESI) [M + H] + 534.3.
步驟2. 在室溫下，向(反式)-N-(8-氨基-7-碘-6-((S)-4-甲基-2-氧代惡唑烷-3-基)噌啉-3-基)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（130mg，0.244mmol）的AcOH（1mL）溶液緩慢加入NCS（N-氯琥珀醯亞胺）（33.0mg，0.244mmol）。將所得混合物在室溫攪拌過夜。將混合物用aq.NaHCO3 （5 mL）稀釋，並用乙酸乙酯（5 mL * 3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓下濃縮。將殘餘物通過prep-TLC（DCM / MeOH = 20/1）純化，得到黃色固體產物（45mg，32.5％產率）。LC-MS（ESI）[M + H]+ 567.8。Step 2. At room temperature, add to (trans)-N-(8-amino-7-iodo-6-((S)-4-methyl-2-oxooxazolidin-3-yl) Lin-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-carboxamide (130mg, 0.244mmol) in AcOH (1mL) was slowly added to NCS (N- Chlorosuccinimide) (33.0mg, 0.244mmol). The resulting mixture was stirred at room temperature overnight. The mixture was diluted with aq. NaHCO 3 (5 mL) and extracted with ethyl acetate (5 mL * 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by prep-TLC (DCM / MeOH = 20/1) to obtain a yellow solid product (45 mg, 32.5% yield). LC-MS (ESI) [M + H] + 567.8.
步驟3. 在氬氣下，向(反式)-N-(8-氨基-5-氯-7-碘-6-((S)-4-甲基-2-氧代惡唑烷-3-基)噌啉-3-基)-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（45.0 mg，0.0794 mmol）和Et3 N（8.00 mg，0.0794 mmol）的EtOAc（3 mL）溶液中加入Pd/C(5.00 mg）。將得到的混合物在室溫和H2 氣氛下攪拌6小時。將反應混合物過濾並將濾液濃縮。通過prep-HPLC（NH3 .H2 O）對粗品進行純化，得到黃色固體產物（8.57mg，24.5％產率）。LC-MS (ESI) [M+H]+ 442.4。1 H NMR (400 MHz, DMSO-d6 ): δ 11.64 (s, 1H), 8.59 (s, 1H), 7.58 (s, 1H), 7.31 (s, 1H), 7.24 (s, 1H), 6.92 (s, 2H), 4.72 – 4.68 (m, 1H), 4.51 – 4.50 (m, 1H), 4.10 – 4.06 (m, 1H), 3.77 (s, 3H), 2.30 – 2.24 (m, 2H), 1.44 – 1.42 (m, 1H), 1.26 – 1.24 (m, 1H), 1.15 (d, J = 6.4 Hz, 3H)。Step 3. Under argon, add (trans)-N-(8-amino-5-chloro-7-iodo-6-((S)-4-methyl-2-oxooxazolidine-3) -Yl) Cinolin-3-yl)-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-carboxamide (45.0 mg, 0.0794 mmol) and Et 3 N (8.00 mg , 0.0794 mmol) in EtOAc (3 mL) was added Pd/C (5.00 mg). The resulting mixture was stirred at room temperature under H 2 atmosphere for 6 hours. The reaction mixture was filtered and the filtrate was concentrated. The crude product was purified by prep-HPLC (NH 3 .H 2 O) to obtain a yellow solid product (8.57 mg, 24.5% yield). LC-MS (ESI) [M+H] + 442.4. 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.64 (s, 1H), 8.59 (s, 1H), 7.58 (s, 1H), 7.31 (s, 1H), 7.24 (s, 1H), 6.92 (s, 2H), 4.72 – 4.68 (m, 1H), 4.51 – 4.50 (m, 1H), 4.10 – 4.06 (m, 1H), 3.77 (s, 3H), 2.30 – 2.24 (m, 2H), 1.44 – 1.42 (m, 1H), 1.26 – 1.24 (m, 1H), 1.15 (d, J = 6.4 Hz, 3H).
2-((8- 氨基 -6-(4- 甲基 -6-( 惡唑 -2- 基 ) 吡啶 -3- 基 ) 噌啉 -3- 基 ) 氨基 )-6- 甲基 -5,6- 二氫 -4H- 吡唑並 [1,5-d] [1,4] 二氮雜卓 -7(8H)- 酮（ 109 ）的製備 
步驟1.根據通用步驟F （在90°C和氬氣氣氛下3小時，以6-氯-8-(((二苯基亞甲基)氨基)噌啉-3-胺和(4-甲基-6-(惡唑-2-基) 吡啶-3-基)硼酸作為反應物），合成了為黃色固體的8-((二苯基亞甲基)氨基)-6-(4-甲基-6-(惡唑-2-基)吡啶-3-基)噌啉-3-胺 （95mg，6.70％產率）。LC-MS（ESI）[M + H]+ 483.1。Step 1. According to general procedure F (at 90°C and argon atmosphere for 3 hours, 6-chloro-8-(((diphenylmethylene)amino)cinoline-3-amine and (4-methyl) Yl-6-(oxazol-2-yl)pyridin-3-yl)boronic acid as the reactant), 8-((diphenylmethylene)amino)-6-(4-methyl) was synthesized as a yellow solid Base-6-(oxazol-2-yl)pyridin-3-yl)cinnoline-3-amine (95 mg, 6.70% yield). LC-MS (ESI) [M + H] + 483.1.
步驟2.根據通用步驟B ，合成了一種呈黃色固體的2-((8-((二苯基亞甲基)氨基)-6-(4-甲基-6-(惡唑-2-基)吡啶-3-基)噌啉-3-基 )氨基)-6-甲基-5,6-二氫-4H-吡唑並[1,5-d] [1,4]二氮雜卓-7(8H)-酮黃色化合物（30 mg，28.1％產率）。LC-MS（ESI）[M + H]+ 646.3。Step 2. According to general procedure B , a yellow solid of 2-((8-((diphenylmethylene)amino)-6-(4-methyl-6-(oxazol-2-yl) was synthesized. )Pyridin-3-yl)Cinnolin-3-yl)amino)-6-methyl-5,6-dihydro-4H-pyrazolo[1,5-d] [1,4]diazepine -7(8H)-one yellow compound (30 mg, 28.1% yield). LC-MS (ESI) [M + H] + 646.3.
步驟3.按照通用步驟C ，合成了一種為黃色固體的2-((8-氨基-6-(4-甲基-6-(惡唑-2-基)吡啶-3-基)噌啉-3-基)氨基)-6 -甲基-5,6-二氫-4H-吡唑並[1,5-d] [1,4]二氮雜卓-7(8H)-酮（109 ）（5.00mg，22.3％產率）。LC-MS (ESI) [M+H]+ 482.1.1 H NMR (400 MHz, DMSO-d6 ): δ 9.87 (s, 1H), 8.57 (s, 1H), 8.33 (s, 1H), 8.09 – 8.05 (m, 2H), 7.49 (s, 1H), 6.90 (s, 1H), 6.57 (s, 3H), 6.05 (s, 1H), 5.00 (s, 2H), 3.85 – 3.82 (m, 2H), 3.08 – 3.06 (m, 2H), 2.95 (s, 3H), 2.42 (s, 3H)。Step 3. According to general procedure C , a yellow solid of 2-((8-amino-6-(4-methyl-6-(oxazol-2-yl)pyridin-3-yl) cinnoline- 3-yl)amino)-6 -methyl-5,6-dihydro-4H-pyrazolo[1,5-d] [1,4]diazepine-7(8H)-one ( 109 ) (5.00 mg, 22.3% yield). LC-MS (ESI) [M+H] + 482.1. 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.87 (s, 1H), 8.57 (s, 1H), 8.33 (s, 1H), 8.09 – 8.05 (m, 2H), 7.49 (s, 1H), 6.90 (s, 1H), 6.57 (s, 3H), 6.05 (s, 1H), 5.00 (s, 2H), 3.85 – 3.82 (m, 2H ), 3.08 – 3.06 (m, 2H), 2.95 (s, 3H), 2.42 (s, 3H).
(E)-2-(2-((8- 氨基 -6-(4- 甲基吡啶 -3- 基 ) 噌啉 -3- 基 ) 氨基 )-6- 甲基 -7- 氧代 -5,6,7, 8- 四氫 -4H- 吡唑並 [1,5-d] [1,4] 二氮雜卓 -1-4- 亞甲基 ) 乙腈（ 110 ）的製備 
步驟1.在0℃下，向（氰甲基）磷酸二乙酯（127 mg，0.72 mmol）的THF（2.00 mL）溶液中添加NaH（24 mg，0.60 mmol），並將其在室溫下攪拌1小時。然後於0°C下，逐滴加入2-溴-6-甲基-5,6-二氫-4H-吡唑並[1,5-d] [1,4]二氮雜卓-4,7(8H)-二酮（103 mg，0.40 mmol）的THF（1.00 mL）溶液，然後在室溫下再攪拌2小時。將混合物用在0°C下用飽和NH4 Cl溶液淬滅，並用EtOAc（5.00 mL x 3）萃取。合併的有機層經無水硫酸鈉乾燥，過濾並濃縮得到粗產物。將殘餘物通過prep-TLC（矽膠，石油醚：乙酸乙酯= 1：3）純化，得到為棕色油的(E)-2-(2-溴-6-甲基-7-氧代-5,6,7,8 -四氫-4H-吡唑並[1,5-d] [1,4]二氮雜卓-4-亞甲基)乙腈（35 mg，產率31.3％）。LC-MS（ESI）[M + H]+ 281.0；[MH]-279.0。Step 1. At 0°C, add NaH (24 mg, 0.60 mmol) to a solution of (cyanomethyl) diethyl phosphate (127 mg, 0.72 mmol) in THF (2.00 mL) and leave it at room temperature Stir for 1 hour. Then at 0°C, dropwise add 2-bromo-6-methyl-5,6-dihydro-4H-pyrazolo[1,5-d] [1,4]diazepine-4, A solution of 7(8H)-dione (103 mg, 0.40 mmol) in THF (1.00 mL) was then stirred at room temperature for another 2 hours. The mixture was quenched with saturated NH 4 Cl solution at 0 °C and extracted with EtOAc (5.00 mL x 3). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The residue was purified by prep-TLC (silica gel, petroleum ether: ethyl acetate = 1:3) to obtain (E)-2-(2-bromo-6-methyl-7-oxo-5) as a brown oil ,6,7,8-Tetrahydro-4H-pyrazolo[1,5-d] [1,4]diazepine-4-methylene)acetonitrile (35 mg, yield 31.3%). LC-MS (ESI) [M + H] + 281.0; [MH]-279.0.
步驟2.按照通用步驟B描述的方法，合成了為黃色固體的(E)-2-(2-(((8-(((二苯基亞甲基)氨基)-6-(4-甲基吡啶-3-基)噌啉-3-基)氨基)- 6-甲基-7-氧代-5,6,7,8-四氫-4H-吡唑並[1,5-d] [1,4]二氮雜卓-4-亞基)乙腈（15 mg，24.4％產率）。LC-MS（ESI）[M + H]+ 616.2；[M-H]- 614.3。Step 2. According to the method described in general step B, (E)-2-(2-(((8-(((diphenylmethylene)amino)-6-(4-methyl) was synthesized as a yellow solid (Pyridin-3-yl) cinnolin-3-yl) amino)-6-methyl-7-oxo-5,6,7,8-tetrahydro-4H-pyrazolo[1,5-d] [1,4] diazepin-4-ylidene) acetonitrile (15 mg, 24.4% yield) .LC-MS (ESI) [ M + H] + 616.2; [MH] - 614.3.
步驟3.按照通用步驟C ，合成了呈黃色固體的(E)-2-(2-(((8-氨基-6-(4-甲基吡啶-3-基)噌啉-3-基)氨基)-6-甲基-7-氧代-5,6,7,8-四氫-4H-吡唑並[1,5-d] [1,4]二氮雜卓-4-亞基)乙腈（110 ）（3.30 mg，％產率）。LC-MS (ESI) [M+H]+ 452.1; [M-H]- 450.2.1 H NMR (400 MHz, CD3 OD): δ 8.57 - 8.53 (m, 2H), 8.12 (s, 1H), 7.69 - 7.67 (m, 1H), 6.92 (s, 1H), 6.66 - 6.64 (m, 2H), 6.49 (s, 1H), 4.80 (s, 2H), 3.80 (s, 2H), 3.23 (s, 2H), 2.50 (s, 3H)。Step 3. According to general procedure C , (E)-2-(2-(((8-amino-6-(4-methylpyridin-3-yl)cinolin-3-yl) was synthesized as a yellow solid Amino)-6-methyl-7-oxo-5,6,7,8-tetrahydro-4H-pyrazolo[1,5-d] [1,4]diazepine-4-subunit ) in acetonitrile (110) (3.30 mg,% yield) .LC-MS (ESI) [ M + H] + 452.1; [MH] - 450.2 1 H NMR (400 MHz, CD 3 OD): δ 8.57 - 8.53. (m, 2H), 8.12 (s, 1H), 7.69-7.67 (m, 1H), 6.92 (s, 1H), 6.66-6.64 (m, 2H), 6.49 (s, 1H), 4.80 (s, 2H) ), 3.80 (s, 2H), 3.23 (s, 2H), 2.50 (s, 3H).
(E)-2-(((8-(E)-2-(((8- 氨基Amino -6-(4--6-(4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-6-)-6- 甲基methyl -4-((1--4-((1- 甲基methyl -1H--1H- 吡唑Pyrazole - 4--4- 基base )) 亞甲基Methylene )-5,6-)-5,6- 二氫Dihydro -4H--4H- 吡唑並Pyrazolo [1,5-d] [1,4][1,5-d] [1,4] 二氮雜卓Diazepam -7(8H)--7(8H)- 酮（ketone( 111111 ）的製備) Preparation
與製備化合物 110 相似的方法合成了為黃色固體的(E)-2- (((8-氨基-6-(4-甲基吡啶-3-基)噌啉-3-基)氨基)-6-甲基-4-((1-甲基-1H-吡唑- 4-基)亞甲基)-5,6-二氫-4H-吡唑並[1,5-d] [1,4]二氮雜卓-7(8H)-酮（111）（7.62mg，25.4％產率）。LC-MS (ESI) [M+H]+ 507.2.1 H NMR (400 MHz, DMSO-d6 ): δ 9.94 (s, 1H), 8.46 - 8.43 (m, 2H), 8.10 (s, 1H), 7.95 (s, 1H), 7.78 (s, 1H), 7.36 (d, J = 5.2 Hz, 1H), 7.03 (s, 1H), 6.84 (s, 1H), 6.59 (s, 1H), 6.55 (s, 3H), 5.13 (s, 2H), 4.69 (s, 2H), 3.89 (s, 3H), 2.90 (s, 3H), 2.33 (s, 3H)。(E)-2-(((8-amino-6-(4-methylpyridin-3-yl)cinolin-3-yl)amino)-6 was synthesized as a yellow solid in a similar method to the preparation of compound 110 -Methyl-4-((1-methyl-1H-pyrazol-4-yl)methylene)-5,6-dihydro-4H-pyrazolo[1,5-d] [1,4 ]Diazepine-7(8H)-one (111) (7.62mg, 25.4% yield). LC-MS (ESI) [M+H] + 507.2. 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.94 (s, 1H), 8.46-8.43 (m, 2H), 8.10 (s, 1H), 7.95 (s, 1H), 7.78 (s, 1H), 7.36 (d, J = 5.2 Hz, 1H ), 7.03 (s, 1H), 6.84 (s, 1H), 6.59 (s, 1H), 6.55 (s, 3H), 5.13 (s, 2H), 4.69 (s, 2H), 3.89 (s, 3H) , 2.90 (s, 3H), 2.33 (s, 3H).
(( 反式Trans )-N-(8-)-N-(8- 氨基Amino -6-((S)-4--6-((S)-4- 甲基methyl -2--2- 氧代惡唑烷Oxoxazolidine -3--3- 基base )) 噌啉Cinolin -3--3- 基base )-2-)-2- 氰基環丙烷Cyanocyclopropane -1--1- 甲醯胺（Formamide ( 112112 ）的製備) Preparation
以與製備化合物 87a / b 類似的方法，合成為黃色固體的(反式)-N-(8-氨基-6-((S)-4-甲基-2-氧代惡唑烷-3-基)噌啉-3-基)- 2-氰基環丙烷-1-甲醯胺（112 ）（3.90 mg，28.5％）。LC-MS (ESI) [M+H]+ 353.4。1 H NMR (400 MHz, DMSO-d6 ) δ 11.75 (s, 1H), 8.40 (s, 1H), 7.28 – 7.27 (m, 1H), 6.95 (d, J = 6.0 Hz, 1H), 6.72 (d, J = 6.8 Hz, 2H), 4.73 – 4.70 (m, 1H), 4.57 (t, J = 8.2 Hz, 1H), 4.09 – 4.06 (m, 1H), 2.86 – 2.82 (m, 1H), 2.20 – 2.16 (m, 1H), 1.67 – 1.62 (m, 1H), 1.48 – 1.44 (m, 1H), 1.31 (d, J = 6.0 Hz, 3H)。In a method similar to the preparation of compound 87a/b , (trans)-N-(8-amino-6-((S)-4-methyl-2-oxooxazolidine-3-) was synthesized as a yellow solid (4) Cinolin-3-yl)-2-cyanocyclopropane-1-carboxamide ( 112 ) (3.90 mg, 28.5%). LC-MS (ESI) [M+H] + 353.4. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.75 (s, 1H), 8.40 (s, 1H), 7.28 – 7.27 (m, 1H), 6.95 (d, J = 6.0 Hz, 1H), 6.72 ( d, J = 6.8 Hz, 2H), 4.73 – 4.70 (m, 1H), 4.57 (t, J = 8.2 Hz, 1H), 4.09 – 4.06 (m, 1H), 2.86 – 2.82 (m, 1H), 2.20 – 2.16 (m, 1H), 1.67 – 1.62 (m, 1H), 1.48 – 1.44 (m, 1H), 1.31 (d, J = 6.0 Hz, 3H).
( 反式 )-N-(8- 氨基 -6-((S)-4- 甲基 -2- 氧代惡唑烷 -3- 基 ) 噌啉 -3- 基 )-1- 甲基 -2-(1- 甲基 - 1H- 吡唑 -4- 基 ) 環丙烷 -1- 甲醯胺（ 113 ）的製備 
步驟1。在室溫下，向4-碘-1-甲基-1H-吡唑（4.50g，21.63mmol）的N，N-二甲基甲醯胺（100.0mL）的溶液中添加DIEA（1.40g，108.0mmol）、甲基丙烯酸乙酯（7.40g，64.9mmol），Pd(PPh3 )2 Cl2 （607mg，0.865mmol）和TBAB（1.40g，4.33mmol）。在Ar下，將混合物在110℃下攪拌16小時。將混合物冷卻至室溫，並用水（100.0mL）稀釋，然後用乙酸乙酯（100.0mL×3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓下濃縮得到粗產物。殘餘物通過柱色譜法（石油醚/乙酸乙酯= 2/1）純化，得到為無色油的(E)-2-甲基-3-(1-甲基-1H-吡唑-4-基)丙烯酸乙酯（1.64g，8.45mmol， 產率39.1%）。LC-MS (ESI) [M+H]+ 195.2。1 H NMR (400 MHz, DMSO-d6 ) δ 8.09 (s, 1H), 7.74 (s, 1H), 7.48 (d, J = 0.8 Hz, 1H), 4.16 (q, J = 7.2 Hz, 2H), 3.87 (s, 3H), 2.01 (s, 3H), 1.22 (t, J = 7.2 Hz, 3H)。step 1. At room temperature, to a solution of 4-iodo-1-methyl-1H-pyrazole (4.50g, 21.63mmol) in N,N-dimethylformamide (100.0mL) was added DIEA (1.40g, 108.0 mmol), ethyl methacrylate (7.40 g, 64.9 mmol), Pd(PPh 3 ) 2 Cl 2 (607 mg, 0.865 mmol) and TBAB (1.40 g, 4.33 mmol). Under Ar, the mixture was stirred at 110°C for 16 hours. The mixture was cooled to room temperature and diluted with water (100.0 mL), and then extracted with ethyl acetate (100.0 mL×3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. The residue was purified by column chromatography (petroleum ether/ethyl acetate = 2/1) to obtain (E)-2-methyl-3-(1-methyl-1H-pyrazol-4-yl) as a colorless oil ) Ethyl acrylate (1.64 g, 8.45 mmol, yield 39.1%). LC-MS (ESI) [M+H] + 195.2. 1 H NMR (400 MHz, DMSO- d6 ) δ 8.09 (s, 1H), 7.74 (s, 1H), 7.48 (d, J = 0.8 Hz, 1H), 4.16 (q, J = 7.2 Hz, 2H), 3.87 (s, 3H), 2.01 (s, 3H), 1.22 (t, J = 7.2 Hz, 3H).
步驟2。在室溫下，向三甲基碘化亞碸（5.24 g，23.8 mmol）的二甲基亞碸（20.0 mL）溶液中加入氫化鈉（722 mg，18.0 mmol）。將混合物在室溫下攪拌1小時。在室溫下向該反應混合物中加入（E）-2-甲基-3-（1-甲基-1H-吡唑-4-基）丙烯酸乙酯（1.40 g，7.22 mmol）的四氫呋喃（5.00 mL）溶液 。將混合物在50℃下攪拌2天。將混合物冷卻至室溫，並用水（100.0mL）稀釋，用乙酸乙酯（100.0mL×3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並在減壓下濃縮，得到粗產物。殘餘物通過柱色譜法（石油醚/乙酸乙酯= 1/1）純化，得到為黃色油的（反式）-1-甲基-2-（1-甲基-1H-吡唑-4-基）環丙烷-1-羧酸乙酯（ 800mg粗品，產率53.3％）。LC-MS（ESI）[M + H] + 209.1。Step 2. At room temperature, add sodium hydride (722 mg, 18.0 mmol) to a solution of trimethyl sulfinium iodide (5.24 g, 23.8 mmol) in dimethyl sulfoxide (20.0 mL). The mixture was stirred at room temperature for 1 hour. To the reaction mixture was added (E)-2-methyl-3-(1-methyl-1H-pyrazol-4-yl)ethyl acrylate (1.40 g, 7.22 mmol) in tetrahydrofuran (5.00 mL) solution. The mixture was stirred at 50°C for 2 days. The mixture was cooled to room temperature, diluted with water (100.0 mL), and extracted with ethyl acetate (100.0 mL×3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. The residue was purified by column chromatography (petroleum ether/ethyl acetate = 1/1) to obtain (trans)-1-methyl-2-(1-methyl-1H-pyrazole-4- Yl) ethyl cyclopropane-1-carboxylate (800 mg crude product, yield 53.3%). LC-MS (ESI) [M + H] + 209.1.
步驟3-6。以與製備化合物 87a / b 類似的方法，合成為黃色固體的(反式)-N-(8-氨基-6-((S)-4-甲基-2-氧代惡唑烷-3-基)噌啉- 3-基)-1-甲基-2-(1-甲基-1H-吡唑-4-基)環丙烷-1-甲醯胺（113 ）（2.24mg，44.2％產率）。LC-MS (ESI) [M+H]+ 422.4。1 H NMR (400 MHz, DMSO-d6 ): δ 10.21 (s, 1H), 8.41 (s, 1H), 7.62 (s, 1H), 7.36 (s, 1H), 7.27 (d, J = 2.0 Hz, 1H), 6.97 (d, J = 2.0 Hz, 1H), 6.68 (s, 2H), 4.75 – 4.70 (m, 1H), 4.60 – 4.56 (m, 1H), 4.10 – 4.06 (m, 1H), 3.82 (s, 3H), 1.70 – 1.68 (m, 1H), 1.33 (d, J = 6.0 Hz, 3H), 1.30 (s, 3H), 1.27 – 1.22 (m, 1H), 1.03 – 1.02 (m, 1H)。Steps 3-6. In a method similar to the preparation of compound 87a/b , (trans)-N-(8-amino-6-((S)-4-methyl-2-oxooxazolidine-3-) was synthesized as a yellow solid Yl) Cinolin-3-yl)-1-methyl-2-(1-methyl-1H-pyrazol-4-yl)cyclopropane-1-carboxamide ( 113 ) (2.24mg, 44.2% yield rate). LC-MS (ESI) [M+H] + 422.4. 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.21 (s, 1H), 8.41 (s, 1H), 7.62 (s, 1H), 7.36 (s, 1H), 7.27 (d, J = 2.0 Hz , 1H), 6.97 (d, J = 2.0 Hz, 1H), 6.68 (s, 2H), 4.75 – 4.70 (m, 1H), 4.60 – 4.56 (m, 1H), 4.10 – 4.06 (m, 1H), 3.82 (s, 3H), 1.70 – 1.68 (m, 1H), 1.33 (d, J = 6.0 Hz, 3H), 1.30 (s, 3H), 1.27 – 1.22 (m, 1H), 1.03 – 1.02 (m, 1H).
N-(8- 氨基 -6-(8- 甲基 -2,3- 二氫 -1H- 吡啶並 [2,3-b] [1,4] 惡嗪 -7- 基 ) 噌啉 -3- 基 )- 2- 氰基 -1- 氟環丙烷 -1- 甲醯胺（ 114 ）的製備 
步驟1. 在室溫下，向2-氰基-1-氟環丙烷-1-羧酸乙酯（240 mg，1.529 mmol）的甲醇（3.00 mL）和水（3.00 mL）溶液中，加入氫氧化鈉（73 mg，1.834 mmol），然後在室溫下攪拌4小時。用1N KHSO4 （水溶液）將混合物的pH調節至約2，然後用乙酸乙酯（10.0mL×3）萃取。合併的有機層用鹽水洗滌，用無水硫酸鈉乾燥，過濾並減壓濃縮，得到為棕色油的2-氰基-1-氟環丙烷-1-羧酸粗品（90mg粗品，產率45.6％），其直接用於下一步驟。LC-MS（ESI）[MH]- 128.1。Step 1. At room temperature, to a solution of ethyl 2-cyano-1-fluorocyclopropane-1-carboxylate (240 mg, 1.529 mmol) in methanol (3.00 mL) and water (3.00 mL), add hydrogen Sodium oxide (73 mg, 1.834 mmol), then stirred at room temperature for 4 hours. The pH of the mixture was adjusted to about 2 with 1N KHSO 4 (aqueous solution), and then extracted with ethyl acetate (10.0 mL×3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain crude 2-cyano-1-fluorocyclopropane-1-carboxylic acid as a brown oil (90mg crude product, yield 45.6%) , Which is used directly in the next step. LC-MS (ESI) [MH ] - 128.1.
步驟2-3。以與製備化合物 87a / b 類似的方法，合成為黃色固體的N-（8-氨基-6-（8-甲基-2,3-二氫-1H-吡啶並[2,3-b] [1,4]惡嗪-7-基）噌啉-3-基）-2-氰基-1-氟環丙烷-1-甲醯胺（114 ）（3.30mg，10.3％收率）。LC-MS (ESI) [M+H]+ 420.2.1 H NMR (400 MHz, CD3 OD): δ 8.62 (s, 1H), 7.39 (s, 1H), 6.95 (s, 1H), 6.83 (s, 1H), 4.38 (t, J = 4.4 Hz, 2H), 3.48 (t, J = 4.4 Hz, 2H), 2.89 - 2.71 (m, 1H), 2.18 - 2.12 (m, 2H), 2.10 (s, 3H)。Step 2-3. In a similar way to the preparation of compound 87a/b , N-(8-amino-6-(8-methyl-2,3-dihydro-1H-pyrido[2,3-b] [ 1,4] Oxazine-7-yl) cinnoline-3-yl)-2-cyano-1-fluorocyclopropane-1-carboxamide ( 114 ) (3.30 mg, 10.3% yield). LC-MS (ESI) [M+H] + 420.2. 1 H NMR (400 MHz, CD 3 OD): δ 8.62 (s, 1H), 7.39 (s, 1H), 6.95 (s, 1H), 6.83 ( s, 1H), 4.38 (t, J = 4.4 Hz, 2H), 3.48 (t, J = 4.4 Hz, 2H), 2.89-2.71 (m, 1H), 2.18-2.12 (m, 2H), 2.10 (s , 3H).
2-((8- 氨基 -6-(8- 甲基 -2,3- 二氫 -1H- 吡啶並 [2,3-b] [1,4] 惡嗪 -7- 基 ) 噌啉 -3- 基 ) 氨基 )-5- 甲基 -4,5- 二氫 -7H- 吡唑並 [5,1-d] [1,2,5] 噻二嗪 6,6- 二氧化物（ 115 ）的製備 
步驟1.根據通用步驟B ，合成為黃色固體的7-(8-(((二苯基亞甲基)氨基)-3-((5-甲基-6,6-二氧代-4,5-二氫-7H-吡唑並[5,1-d] [1,2,5] 噻二嗪-2-基)氨基)噌啉-6-基)-8-甲基-2,3-二氫-1H-吡啶並 [2,3-b] [1, 4]惡嗪-1-甲酸叔丁基酯（6.67mg，25.1％產率）。LC-MS（ESI）[M + H]+ 758.2。Step 1. According to general procedure B , 7-(8-(((diphenylmethylene)amino)-3-((5-methyl-6,6-dioxo-4, 5-Dihydro-7H-pyrazolo[5,1-d] [1,2,5]thiadiazine-2-yl)amino)cinnoline-6-yl)-8-methyl-2,3 -Dihydro-1H-pyrido[2,3-b] [1, 4] oxazine-1-carboxylic acid tert-butyl ester (6.67 mg, 25.1% yield). LC-MS (ESI) [M + H ] + 758.2.
步驟2.根據通用步驟E ，合成為黃色固體的2-((8-氨基-6-(8-甲基-2,3-二氫-1H-吡啶並[2,3-b] [1,4]惡嗪-7-基)噌啉-3-基)氨基)-5-甲基-4,5-二氫-7H-吡唑並 [5,1-d] [1,2,5]噻二嗪6,6-二氧化物（115 ）（5.47mg，56.0％產率）。LC-MS (ESI) [M+H]+ 494.6。1 H NMR (400 MHz, DMSO-d6 ) δ 10.05 (s, 1H), 7.97 (s, 1H), 7.35 (s, 1H), 6.64 (s, 1H), 6.51 (s, 3H), 6.23 (s, 1H), 5.65 – 5.63 (m, 3H), 4.62 (s, 2H), 4.27 (s, 2H), 3.38 – 3.36 (m, 2H), 2.88 (s, 3H), 2.04 (s, 3 H)。Step 2. According to general procedure E , 2-((8-amino-6-(8-methyl-2,3-dihydro-1H-pyrido[2,3-b] [1, 4] Oxazine-7-yl) cinnoline-3-yl) amino)-5-methyl-4,5-dihydro-7H-pyrazolo[5,1-d] [1,2,5] Thiadiazine 6,6-dioxide ( 115 ) (5.47 mg, 56.0% yield). LC-MS (ESI) [M+H] + 494.6. 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.05 (s, 1H), 7.97 (s, 1H), 7.35 (s, 1H), 6.64 (s, 1H), 6.51 (s, 3H), 6.23 ( s, 1H), 5.65 – 5.63 (m, 3H), 4.62 (s, 2H), 4.27 (s, 2H), 3.38 – 3.36 (m, 2H), 2.88 (s, 3H), 2.04 (s, 3 H) ).
2-((8-2-((8- 氨基Amino -6-(5--6-(5- 氨基Amino -4--4- 甲基吡啶Picoline -3--3- 基base )) 噌啉Cinolin -3--3- 基base )) 氨基Amino )-5-)-5- 甲基methyl -4,5--4,5- 二氫Dihydro -7H--7H- 吡唑並Pyrazolo [5, 1-d] [1,2,5][5, 1-d] [1,2,5] 噻二嗪Thiadiazine 6,6-6,6- 二氧化物（Dioxide ( 116116 ）的製備) Preparation
以與製備化合物 115 類似的方法，合成為黃色固體的2-((8-氨基-6-(5-氨基-4-甲基吡啶-3-基)噌啉-3-基)氨基)-5-甲基-4,5 -二氫-7H-吡唑並[5,1-d] [1,2,5]噻二嗪6,6-二氧化物（116 ）（4.36mg，0.00967mmol，39.5％產率）。LC-MS (ESI) [M+H]+ 452.5。1 H NMR (400 MHz, DMSO-d6 ) δ 10.06 (s, 1H), 7.99 (s, 1H), 7.95 (s, 1H), 7.67 (s, 1H), 6.66 (s, 1H), 6.54 – 6.51 (m, 3H), 6.23 (s, 1H), 5.62 (s, 2H), 5.22 (s, 2H), 4.62 (s, 2H), 2.88 (s, 3H), 2.02 (s, 3 H)。In a method similar to the preparation of compound 115 , 2-((8-amino-6-(5-amino-4-methylpyridin-3-yl)cinolin-3-yl)amino)-5 was synthesized as a yellow solid -Methyl-4,5-dihydro-7H-pyrazolo[5,1-d] [1,2,5]thiadiazine 6,6-dioxide ( 116 ) (4.36mg, 0.00967mmol, 39.5% yield). LC-MS (ESI) [M+H] + 452.5. 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.06 (s, 1H), 7.99 (s, 1H), 7.95 (s, 1H), 7.67 (s, 1H), 6.66 (s, 1H), 6.54 – 6.51 (m, 3H), 6.23 (s, 1H), 5.62 (s, 2H), 5.22 (s, 2H), 4.62 (s, 2H), 2.88 (s, 3H), 2.02 (s, 3 H).
生物學評價Biological evaluation
為評估化合物對HPK-1的抑制，對式（I）或（Ia）的示例性化合物進行了測試。通過結合或ADP-Glo試驗測定每個示例性化合物的Ki或IC50。In order to evaluate the inhibition of HPK-1 by compounds, the exemplary compounds of formula (I) or (Ia) were tested. The Ki or IC50 of each exemplary compound was determined by binding or ADP-Glo test.
[生化結合試驗][Biochemical Combination Test]
1、試驗原理1. Test principle
LanthaScreen®Eu激酶結合試驗是基於一個專有物（Alexa Fluor®647標記的）、即ATP競爭性的激酶結構（激酶示蹤劑）同HPK1的競爭性結合和置換。使用能與HPK1結合的抗標籤抗體來檢測示蹤劑與激酶的結合。當示蹤劑和抗體同時與HPK-1結合時，會導致銪（eu）供體螢光團到激酶示蹤劑上Alexa fluor®647受體螢光團的高度的FRET（螢光共振能量轉移）。一個抑制劑與HPK-1結合，會與示蹤劑競爭，導致FRET的丟失。LanthaScreen®Eu kinase binding test is based on a proprietary (Alexa Fluor®647-labeled), namely ATP-competitive kinase structure (kinase tracer) competitive binding and replacement with HPK1. An anti-tag antibody capable of binding to HPK1 is used to detect the binding of the tracer to the kinase. When the tracer and antibody bind to HPK-1 at the same time, it will cause a high degree of FRET (fluorescence resonance energy transfer) from the europium (eu) donor fluorophore to the Alexa fluor®647 acceptor fluorophore on the kinase tracer. ). An inhibitor that binds to HPK-1 will compete with the tracer, resulting in the loss of FRET.
2、材料 白色ProxiPlate 384F（測試板）（White ProxiPlate 384F，PerkinElmer #6008289）； 384微孔板（化合物板）（ Corning #3701)； HPK-1酶（Singalchem-m23-11G） 示蹤劑222（Tracer 222，Invitrogen-PV6121） Eu-抗-GST抗體（Invitrogen-PV5594） 測試緩衝液：2mM DTT，0.01% BRIJ-35，100 mM MgCl2 ，50 mM HEPES2. Material: White ProxiPlate 384F (test plate) (White ProxiPlate 384F, PerkinElmer #6008289); 384 microplate (compound plate) (Corning #3701); HPK-1 enzyme (Singalchem-m23-11G) tracer 222 ( Tracer 222, Invitrogen-PV6121) Eu-anti-GST antibody (Invitrogen-PV5594) Test buffer: 2mM DTT, 0.01% BRIJ-35, 100 mM MgCl 2, 50 mM HEPES
3、方法3. Method
3.1 平板佈局和化合物稀釋3.1 Plate layout and compound dilution
製備濃度為5 mM（100X）的測試化合物的儲備溶液。在384孔板中的第2-13列中加入12.5μL /孔的儲備溶液，在第3-12列、14-23列、孔A1-H1、I24-P24孔中加入10μL /孔的DMSO。Prepare a stock solution of the test compound at a concentration of 5 mM (100X). Add 12.5μL/well of stock solution to column 2-13 of the 384-well plate, and add 10μL/well of DMSO to column 3-12, column 14-23, wells A1-H1, and I24-P24.
對於參考化合物，最高濃度為1 mM。在J1-P1和A24-H24孔中加入10ul/孔2 mM的葡萄球菌素。所有濃度均以5倍的梯度往下稀釋,在384孔板的第2列和第13列加入2.5 uL的溶液，第3列和第14列加入10 uL的 DMSO。For the reference compound, the highest concentration is 1 mM. Add 10ul/well of 2 mM Staphylococcus to wells J1-P1 and A24-H24. All concentrations are diluted down in a 5-fold gradient. Add 2.5 uL of the solution in the second and 13th columns of the 384-well plate, and add 10 uL of DMSO in the third and 14th columns.
將384孔板以2500轉/分的速度離心1分鐘,將80uL的化合物溶液轉移到測試板中,一個稀釋板做兩個測試板。Centrifuge the 384-well plate at a speed of 2500 rpm for 1 minute, and transfer 80 uL of the compound solution to the test plate. One dilution plate is used as two test plates.
3.2 檢測條件和參數：3.2 Testing conditions and parameters:
2nM HPK-1、2Nm Eu抗GST抗體、15nM示蹤劑222，孵育時間60分鐘。2nM HPK-1, 2Nm Eu anti-GST antibody, 15nM tracer 222, incubation time 60 minutes.
TR-FRET，340nm ex/615nm和665nm em；100 usee延遲（delay）；和200 usee 積分（integration）。TR-FRET, 340nm ex/615nm and 665nm em; 100 usee delay (delay); and 200 usee integration (integration).
3.3 結合試驗3.3 Combination test
將4μL 2X HPK-1和Eu-抗-GST抗體添加到測試板的每個孔中。將溶液在23℃培養箱中培養1h。向每孔加入4μL 2X示蹤劑-222，再在23℃培養箱中培養lh。Add 4 μL of 2X HPK-1 and Eu-anti-GST antibody to each well of the test plate. The solution was incubated in an incubator at 23°C for 1 h. Add 4μL of 2X Tracer-222 to each well, and then incubate in an incubator at 23°C for 1h.
3.4資料分析：3.4 Data analysis:
用Morrison-Ki-Fit模型對化合物K進行了XL-Fit分析。 fit = (l-((((E+x)+(Ki*(l+(S/Kd))))-(((((E+x＞+-(Ki*(l+(S/Kd)))) 2) -((4* E)*c)) ^0.5))/(2* E)))The Morrison-Ki-Fit model was used to analyze compound K by XL-Fit. fit = (l-((((E+x)+(Ki*(l+(S/Kd))))-((((((E+x＞+-(Ki*(l+(S/Kd)) )) 2) -((4* E)*c)) ^0.5))/(2* E)))
E =酶濃度， S=示蹤劑222濃度， Kd = 示蹤劑222 KdE = enzyme concentration, S = tracer 222 concentration, Kd = tracer 222 Kd
[ADP-Glo實驗][ADP-Glo experiment]
1.材料與儀器 Tris（Sinopharm）， 氯化鎂，DTT（Sigma） HPK1酶（Carna），BSA（PE） ADP-Glo激酶檢測試劑盒，ATP（Promega） 384聚苯乙烯淺平底白板（Greiner） Biotek讀板器（Biotek） 微孔板低速離心機（湘智）1. Materials and Instruments Tris (Sinopharm), Magnesium chloride, DTT (Sigma) HPK1 enzyme (Carna), BSA (PE) ADP-Glo Kinase Detection Kit, ATP (Promega) 384 polystyrene shallow flat white board (Greiner) Biotek Plate Reader (Biotek) Microplate Low Speed Centrifuge (Xiangzhi)
2.方法2. Method
2.1溶液製備2.1 Solution preparation
a. 試驗緩衝液：ddH2 O中含Tris（40mM），MgCl2 （20mM），BSA（0.1％），DTT（0.5mM）a. Test buffer: Tris (40mM), MgCl 2 (20mM), BSA (0.1%), DTT (0.5mM) in ddH 2 O
b. 激酶溶液：1X試驗緩衝液中的HPK-1（0.2ng /μL）b. Kinase solution: HPK-1 in 1X assay buffer (0.2ng/μL)
c. 底物溶液：1X試驗緩衝液中的ATP（2μM）c. Substrate solution: ATP (2μM) in 1X assay buffer
d. 抑制劑溶液：從1000nM開始，稀釋3倍， 12個濃度點。d. Inhibitor solution: starting from 1000nM, diluted 3 times, 12 concentration points.
2.2試驗方案2.2 Test plan
a. 將測試化合物和參考化合物儲備溶液（10mM）用100％DMSO稀釋50倍。在96孔稀釋板上對4X系列測試化合物用DMSO進行稀釋。將1μL稀釋的抑制劑溶液與49μL試驗緩衝液混合；a. Dilute the test compound and reference compound stock solution (10mM) with 100% DMSO 50 times. The 4X series of test compounds were diluted with DMSO on a 96-well dilution plate. Mix 1μL of diluted inhibitor solution with 49μL of test buffer;
b. 震動20分鐘；b. Shake for 20 minutes;
c. 將2μL HPK-1與1μL抑制劑溶液一起添加至384孔測定板中。1000 rpm / min 1分鐘，在25℃孵育10分鐘。c. Add 2μL HPK-1 and 1μL inhibitor solution to the 384-well assay plate. 1000 rpm/min for 1 minute, incubate at 25°C for 10 minutes.
d. 將1μL底物溶液加入至試驗體系中， 1000rpm / min 1分鐘，在25℃孵育60分鐘。d. Add 1μL of substrate solution to the test system, 1000rpm/min for 1 minute, and incubate at 25°C for 60 minutes.
e.加入4μL ADP-Glo試劑， 1000 rpm / min 1min，在25℃下孵育40 min。e. Add 4μL of ADP-Gloreagent, 1000 rpm / min for 1 min, and incubate at 25°C for 40 min.
f. 將8μL檢測試劑加入至試驗體系中，1000rpm / min 1min，在25℃下孵育40分鐘f. Add 8μL of detection reagent to the test system, 1000rpm / min for 1min, and incubate at 25℃ for 40 minutes
g.利用讀板器讀取讀取器RLU（相對發光單位）信號。g. Use the plate reader to read the RLU (Relative Luminescence Unit) signal of the reader.
表2 不同化合物的IC50
值
在Jurkat細胞功能試驗和Pan T細胞功能試驗中測試了式（I）或（Ia）的示例性化合物。Exemplary compounds of formula (I) or (Ia) were tested in the Jurkat cell function test and the Pan T cell function test.
[Jurkat細胞中細胞功能試驗][Cell function test in Jurkat cells]
1. 材料與儀器
2. 細胞系和細胞培養2. Cell lines and cell culture
Jurkat E6-1細胞系（Cat＃SCSP-513）購自中國科學院。在完全培養基（90％RPMI培養基，10％FBS）中培養細胞。細胞每週約傳三次，並保持在1×105 細胞/ mL至1×106 細胞/ mL的融合度。Jurkat E6-1 cell line (Cat#SCSP-513) was purchased from the Chinese Academy of Sciences. Culture the cells in complete medium (90% RPMI medium, 10% FBS). Cells are transferred approximately three times a week and maintained at a confluency of 1×10 5 cells/mL to 1×10 6 cells/mL.
3. 測試化合物的工作溶液製備3. Preparation of working solution for test compound
3.1首先將測試化合物溶解在DMSO中 ，最終濃度為10mM，作為儲備液。3.1 First Test compounds were dissolved in DMSO, final concentration of 1OmM, as a stock solution.
3.2用DMSO按1：3的比例對每種化合物的儲備液進行系列稀釋，以製備另外的6種中間溶液，包括7個點；3.2 Use DMSO to serially dilute the stock solution of each compound in a ratio of 1:3 to prepare another 6 intermediate solutions, including 7 points;
3.3在測定之前，通過使用完全培養基（最終濃度的4倍）將中間溶液稀釋250倍來製備工作溶液3.3 Before the measurement, prepare the working solution by diluting the intermediate solution 250 times with complete medium (4 times the final concentration)
4. 實驗步驟4. Experimental steps
4.1對細胞計數並測定活細胞密度，用溫育培養基稀釋細胞至工作細胞密度為1×106 活細胞/ mL。4.1 Count the cells and determine the density of viable cells. Dilute the cells with incubation medium to a working cell density of 1×10 6 viable cells/mL.
4.2移取100μL Jurkat細胞懸液至96孔培養板的每個孔中。4.2 Pipette 100 μL of Jurkat cell suspension into each well of a 96-well culture plate.
4.3吸取50μL測試化合物工作溶液至96孔培養板的各孔中以開始反應。將板放回培養箱1小時。4.3 Pipette 50 μL of the test compound working solution into each well of the 96-well culture plate to start the reaction. Return the plate to the incubator for 1 hour.
4.4稀釋ImmunoCult Hu CD3 / CD28 T細胞啟動劑：將100μL儲備溶液添加到900μL完全培養基中，然後將50μL T細胞啟動劑工作溶液移取到96孔培養板的各個孔中以開始反應。將板放回培養箱24小時。4.4 Dilute ImmunoCult Hu CD3/CD28 T cell activator: add 100μL of stock solution to 900μL of complete medium, and then transfer 50μL of T cell activator working solution to each well of a 96-well culture plate to start the reaction. Return the plate to the incubator for 24 hours.
4.5孵育24小時後，提取100μL培養上清液用於IFN-β檢測。4.5 After 24 hours of incubation, 100 μL of culture supernatant was extracted for IFN-β detection.
5.資料分析5. Data analysis
進行所有計算，並通過Graphpadprism V8確定IC50。Perform all calculations and determine IC50 with Graphpadprism V8.
[CD3+ T細胞的細胞功能試驗][Cell function test of CD3 + T cells]
1.材料和儀器
2. 製備CD3+ 細胞2. Preparation of CD3 + cells
2.1冷凍保存的PB CD3 + 總T細胞（Cat＃SLB-CD3T-10B）購自Saily Bio，並保存在液氮中。2.1 The cryopreserved PB CD 3 + total T cells (Cat#SLB-CD3T-10B) were purchased from Saily Bio and stored in liquid nitrogen.
2.2使用前準備新鮮的完全培養基：將4μL rhIL-2加入到ImmunoCult-XF T細胞Exp培養基中（rhIL-2：32.8 IU / ml，800pg / ml）。2.2 Prepare fresh complete medium before use: add 4μL rhIL-2 to ImmunoCult-XF T cell Exp medium (rhIL-2: 32.8 IU/ml, 800pg/ml).
2.3準備CD3 + 細胞：在37°C水浴中完全孵育培養基，使用前至少加熱15分鐘。從儲存器中轉移一瓶冷凍保存的T細胞，確保小瓶保持低溫，直到解凍過程得以確保。將小瓶放入37°C水浴中解凍細胞，然後輕輕搖動小瓶2分鐘。解凍完成後，用70％乙醇噴灑小瓶，然後將小瓶移至生物安全櫃中。使用寬口移液器吸頭將T細胞轉移到裝有完全培養基的25 mL錐形管中。通過將50 mL錐形管放入離心機中以1200 rpm旋轉10分鐘來計數細胞。旋轉完成後，吸出培養基並將T細胞以1×106 細胞/ mL的密度重新懸浮在足夠的孵育的培養基中，將細胞轉移到T25細胞培養皿中，然後將平板放回培養箱中1小時。2.3 Prepare CD 3 + cells: Completely incubate the medium in a 37°C water bath and heat it for at least 15 minutes before use. Transfer a vial of cryopreserved T cells from the reservoir, ensuring that the vial is kept cold until the thawing process is ensured. Place the vial in a 37°C water bath to thaw the cells, then gently shake the vial for 2 minutes. After thawing is complete, spray the vial with 70% ethanol, and then move the vial to the biological safety cabinet. Use a wide-mouth pipette tip to transfer the T cells to a 25 mL conical tube containing complete medium. The cells were counted by placing a 50 mL conical tube in a centrifuge and spinning at 1200 rpm for 10 minutes. After the rotation is complete, aspirate the medium and resuspend the T cells in sufficient incubation medium at a density of 1×10 6 cells/mL, transfer the cells to a T25 cell culture dish, and then put the plate back in the incubator for 1 hour .
3.測試化合物的工作溶液製備3. Preparation of working solution of test compound
3.1首先將測試化合物溶解在DMSO中，最終濃度為10mM，作為儲備液。3.1 First, the test compound was dissolved in DMSO, the final concentration was 10 mM, as a stock solution.
3.2用DMSO按1：3的比例對每種化合物的儲備液進行系列稀釋，以製備另外的6種中間溶液，包括7個點。3.2 The stock solution of each compound was serially diluted in a ratio of 1:3 with DMSO to prepare another 6 kinds of intermediate solutions, including 7 points.
3.3測定前，使用完全培養基（4x終濃度）將中間溶液稀釋250倍來製備工作溶液。3.3 Before the measurement, use complete medium (4x final concentration) to dilute the intermediate solution 250 times to prepare a working solution.
4. 實驗步驟：4. Experimental steps:
4.1移取100μL T細胞懸液至96孔培養板的每個孔中。4.1 Pipette 100 μL of T cell suspension into each well of a 96-well culture plate.
4.2吸取50μL測試化合物工作溶液到96孔培養板的各孔中以開始反應。將板放回培養箱1小時。4.2 Pipette 50 μL of the test compound working solution into each well of the 96-well culture plate to start the reaction. Return the plate to the incubator for 1 hour.
4.3稀釋ImmunoCult Hu CD3 / CD28 T細胞啟動劑，將100μL儲備溶液添加到900μL完全培養基中，然後將50μL T細胞啟動劑的工作溶液移取到96孔培養板的各個孔中以開始反應。將板放回培養箱24小時。4.3 Dilute the ImmunoCult Hu CD3/CD28 T cell activator, add 100 μL of stock solution to 900 μL of complete medium, and then transfer 50 μL of the working solution of the T cell activator to each well of the 96-well culture plate to start the reaction. Return the plate to the incubator for 24 hours.
4.4孵育24小時後，提取100μL細胞培養上清液，用於IFN-β檢測。4.4 After 24 hours of incubation, extract 100 μL of cell culture supernatant for IFN-β detection.
5.資料分析5. Data analysis
進行所有計算，並通過Graphpad prism V8確定IC50。Emax =相較於未處理細胞，IL-2增加的最大百分比。Perform all calculations and determine IC50 with Graphpad prism V8. Emax = the maximum percentage increase in IL-2 compared to untreated cells.
表3
肝細胞的代謝穩定性Metabolic stability of liver cells
將本文所述的化合物與來自人、猴子、犬、大鼠和小鼠的肝細胞以0.5×106 個活細胞/ mL的細胞密度孵育。該孵育在化合物濃度為1μM下進行。在0、15、30、60、90和120分鐘時取樣，並與混有內標的乙腈混合以終止反應。對淬滅的樣品進行離心，並使用LC/ MS/MS分析上清液，以測定每個時間點剩餘的化合物。從化合物消失開始計算t1/2 。在某些實施方式中，在表4中描述了所測定的化合物87b在不同物種中的t1/2 和體外清除率（Cl）。根據測定的代謝穩定性t1/2 ，應用一種常用方法（Obach RS（1999），Drug Metabolism and Disposition 27，1350 – 1359）計算體外CL。The compounds described herein were incubated with hepatocytes from humans, monkeys, dogs, rats, and mice at a cell density of 0.5×10 6 viable cells/mL. The incubation was performed at a compound concentration of 1 μM. Samples were taken at 0, 15, 30, 60, 90, and 120 minutes, and mixed with acetonitrile mixed with internal standard to terminate the reaction. The quenched samples were centrifuged, and the supernatant was analyzed using LC/MS/MS to determine the remaining compounds at each time point. Calculate t 1/2 from the disappearance of the compound. In some embodiments, Table 4 describes the determined t 1/2 and in vitro clearance (Cl) of compound 87b in different species. Based on the determined metabolic stability t 1/2 , a common method (Obach RS (1999), Drug Metabolism and Disposition 27, 1350 – 1359) was used to calculate the in vitro CL.
表4 化合物 87b在肝細胞的代謝穩定性
藥代動力學Pharmacokinetics
將在5％DMSO / 40％PEG400 / 55％水中製備的0.2 mg / mL的本文所述化合物的給藥溶液，以1mg/kg通過靜脈內（IV）推注給藥於一組2-3只雄性CD-1小鼠（4-6周大，20-30g）。在給藥後0.033、0.083、0.25、0.5、1、2、4、8和24小時，通過頸靜脈將血液樣品收集到含有肝素鈉作為抗凝劑的試管中。然後將血液樣品離心，並使用LC/MS /MS對所得血漿樣品進行分析以確定化合物的濃度。使用非房室模型來計算藥代動力學（PK）參數。在某些實施方式中，本文表5中描述了化合物在小鼠中的藥代動力學。A 0.2 mg/mL dosing solution of the compound described herein, prepared in 5% DMSO / 40% PEG400 / 55% water, was administered to a group of 2-3 rats by intravenous (IV) bolus injection at 1 mg/kg Male CD-1 mice (4-6 weeks old, 20-30g). At 0.033, 0.083, 0.25, 0.5, 1, 2, 4, 8 and 24 hours after administration, blood samples were collected via the jugular vein into test tubes containing heparin sodium as an anticoagulant. The blood sample is then centrifuged, and the resulting plasma sample is analyzed using LC/MS/MS to determine the concentration of the compound. A non-compartmental model is used to calculate pharmacokinetic (PK) parameters. In certain embodiments, the pharmacokinetics of the compounds in mice are described in Table 5 herein.
表5 小鼠藥代動力學
無no
無no
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| WO2019089835A1 (en) * | 2017-10-31 | 2019-05-09 | Samumed, Llc | Diazanaphthalen-3-yl carboxamides and preparation and use thereof |
| JP7433304B2 (en) * | 2018-09-30 | 2024-02-19 | エフ・ホフマン－ラ・ロシュ・アクチェンゲゼルシャフト | Cinnoline compounds and the treatment of HPK1-dependent disorders such as cancer |
-
2020
- 2020-07-10 WO PCT/CN2020/101409 patent/WO2021004535A1/en active Application Filing
- 2020-07-10 TW TW109123343A patent/TW202116753A/en unknown
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| Publication number | Publication date |
|---|---|
| WO2021004535A1 (en) | 2021-01-14 |
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