TW201228664A

TW201228664A - Pyrrolo-nitrogenous heterocyclic derivatives pharmaceutical salts, preparation process and pharmaceutical use thereof

Info

Publication number: TW201228664A
Application number: TW100112930A
Authority: TW
Inventors: Peng-Cho Tang
Original assignee: Jiangsu Hengrui Medicine Co; Shanghai Hengrui Pharm Co Ltd
Priority date: 2011-01-07
Filing date: 2011-04-14
Publication date: 2012-07-16

Abstract

The present disclosure relates to pyrrolo-nitrogenous heterocyclic derivatives pharmaceutical salts, preparation processes and pharmaceutical use thereof. Specifically, the present disclosure relates to (R, Z)-2-(5-fluoro-2-oxo-1, 2-dihydro-indol-3-ylidenemethyl)-5-(2-hydroxy-3-morphol in-4-yl-propyl)-3-methyl-5, 6, 7, 8-tetrahydro-1H-pyrrolo[3, 2-c]azepin-4-one derivatives pharmaceutical salts presented by formula (I), the uses for treatment especially for protein kinase inhibitors.

Description

201228664 六、發明說明：【發明所屬之技術領域】 ,本發明涉及吡咯並N雜環類衍生物的可藥用的_、其製備方法及其在醫藥上的應用，特別是（尤幻_2〜（5~氟侧氧基-1，2-二氫-吲哚-3-次曱基）-5_(2-羥基-3—嗎啉〜4 基-丙基）-3-甲基一5, 6, 7, 8-四氫-1万-吡咯並[3, 2〜c]吖庚因-4-酮的可藥用鹽及其製備方法，以及作為治療劑特別是作為蛋白激酶抑制劑的用途。【先前技術】 k號傳導作為細胞的一種基礎調節機制將胞外的各種信號傳遞到細胞内部，使細胞做出應答，實現諸如增殖、分化、凋亡等過程。蛋白激酶（PKs)在這一過程中有著重要作用。PKs可分為酪胺酸激酶（PTKs)和絲胺酸/蘇胺酸激酶 (STKs)。PTKs可使蛋白質上的酪胺酸殘基磷酸化，Sns 可磷酸化絲胺酸、蘇胺酸殘基。酪胺酸激酶又可分為受體型（receptor tyrosine kinase，RTKs)和非受體型 (non-receptor tyrosine kinase)。 RTKs家族又可劃分為許多亞族，主要包括（ι)Ε^Β (Her)家族’包括 EGFROler-l)、Her-2、Her-3、Her一4 ; (2)胰島素受體家族，包括胰島素受體IR、姨島素樣生長因子I受體⑽1R)f ;⑶ΠΙ型家族，包括血小板衍生生長因子受體PDGFR，幹細胞因子SCFR (c—Kit)等。此外，肝細胞生長因子受體e-Met，血㈣皮生長因子受體νΕ· 等也屬於RTKs家族。它們作為信號傳遞者在調節細胞增殖 95166 4 201228664 和分化凋零方面均起著關鍵作用。（Schlessinger and201228664 VI. Description of the invention: [Technical field to which the invention pertains] The present invention relates to a pharmaceutically acceptable form of a pyrrolo-N heterocyclic derivative, a preparation method thereof and its use in medicine, in particular, ~(5~Fluoro-oxy-1,2-dihydro-indole-3-indenyl)-5-(2-hydroxy-3-morpholine~4-yl-propyl)-3-methyl-5 , 6, 7, 8-tetrahydro-10,000-pyrrolo[3,2~c]azepin-4-one pharmaceutically acceptable salt, a process for the preparation thereof, and as a therapeutic agent, particularly as a protein kinase inhibitor [Prior Art] K-conduction is a basic regulatory mechanism of cells that transmits extracellular signals to the interior of cells, allowing cells to respond to processes such as proliferation, differentiation, and apoptosis. Protein kinases (PKs) It plays an important role in this process. PKs can be divided into tyrosine kinases (PTKs) and serine/threonine kinases (STKs). PTKs can phosphorylate tyrosine residues on proteins, Sns phosphoric acid Residues of serine and threonine. Tyrosine kinases can be further divided into receptors (receptor tyrosine kinases, RTKs) and non-receptor tyrosines. The RTKs family can be divided into many subfamilies, including (I) Β^Β (Her) family 'including EGFROler-l), Her-2, Her-3, Her-4; (2) insulin receptor Family, including insulin receptor IR, 姨-like growth factor I receptor (10) 1R) f; (3) ΠΙ-type family, including platelet-derived growth factor receptor PDGFR, stem cell factor SCFR (c-Kit) and the like. In addition, the hepatocyte growth factor receptor e-Met, blood (tetra) dermatodermal growth factor receptor νΕ·, etc. also belong to the RTKs family. They act as signal transmitters in regulating cell proliferation 95166 4 201228664 and differentiation and dying. (Schlessinger and

Ullrich,此i/ro/? 1992，9，383)。 EGFR亞族是RTKs家族的重要的成員之一，該家族受體藉由配體介導的受體間同型或異型二聚化過程完成啟動’二聚化使受體胞内催化區的酪胺酸殘基磷酸化，並可作為後續信號分子的結合位元點進而啟動諸如絲裂原啟動蛋白激酶（MAP激酶）和磷脂醯肌醇激酶（ρι_3激酶）等胞内信號級聯’最終實現細胞對信號的應答。在大部分人類實體腫瘤中，如乳腺癌、***癌、非小細胞肺癌、胃腸癌、卵巢癌、胰腺癌等’均存在EGFR和/或Her-2發生突變而異常活化或過度表達的情況，從而使其與腫瘤發生發展的關聯性得到了確認。 RTKs Class III亞族的血小板衍生生長因子受體 (PDGFR)和c-Kit，與ErbB家族類似，也藉由二聚化過程活化後傳遞信號。該家族成員與腫瘤細胞的分化，增殖遷移以及血管生成過程密切相關。例如在小細胞支氣管癌、黑素瘤、乳腺癌、成神經細胞瘤中，c_Kit都存在高度表達或犬釔（參見 Schii tte et al.，innovartis 3/2001 )。尤其在疋月腸道膠質瘤（GIST)，基因突變可使c_Kit受體持續活化，促使細胞***率增加，進而導致基因組不穩定誘發癌變。（參見 weber 等，乂 ^c〇7· 22(14S)， 9642 (2004)) RTKs的另一重要成員是血管内皮生長因子受體（VEGFR)。 VEGFR與血管生成過程直接相關，它能夠誘導内皮細胞的 95166 5 201228664 增殖和遷移’促進毛細血管生成，形成超滲透不成熟的血管網路，為腫瘤生長提供營養◊除了促血管生成活性，VEGFR 及VEGF可在腫瘤細胞内直接藉由pr〇 survivai機制促進腫瘤生長。藉由研究發現卩阢叩在各種惡性實體腫瘤中，如肺癌、乳腺癌、卵巢癌、胰腺癌和黑素瘤中均有大量表達，因此藉由抑制VEGFR活性而實現抑制腫瘤生長對於腫瘤治療有很大的應用價值。此外，作為RTKs家族成員的肝細胞生長因子受體 c-Met(HGFR)，經文獻證實其與腫瘤生成、侵襲和轉移，細胞運動性增強等也密切相關（參見Ma，P. C等（2〇〇3b) 22，309-25; Maulik，G·等 (2002b). 〇^〇々/此以⑽仏13，41_59)。腫瘤細胞的主要特徵是基因組損傷和信號調節通路的失控。基因組損傷導致部分關鍵調節蛋白的生物功能發生改變或喪失，進而使信號傳導過程遭到破壞，異常的产號通路使得腫瘤細胞能夠在基因損傷的狀態下繼續存活和增殖。作為實現這些調節過程的根本，PTKs與腫'瘤的發生和發展密切相關，因而成為重要的腫瘤治療靶點。人們希望藉由抑制RTKs中的一種或者多種，有效地改善和治療由 RTKs介導的細胞非正常增殖而造成的生理奈亂。 W02008/138232中公開了一類新型的π比p各並n雜學類衍生物及其作為蛋白激酶抑制劑的應用，其中公開的實施例53為式（I)所示的（疋^)-2-(5-氟-2-侧氧基-1，2、二氣吲哚-3-次曱基）-5-(2-羥基-3-嗎啉-4-基-丙基）-3〜甲烏 95166 201228664 -5, 6, 7, 8-四氫-1及-吡咯並[3, 2-c]吖庚因_4_酮。Ullrich, this i/ro/? 1992, 9, 383). The EGFR subfamily is an important member of the RTKs family, which initiates the 'dimerization of the receptor's intracellular catalytic region of tyramine by ligand-mediated inter-receptor homotypic or heterodimerization. Phosphorylation of acid residues and acting as a binding site for subsequent signaling molecules to initiate intracellular signaling cascades such as mitogen-activated protein kinase (MAP kinase) and phospholipidinokinase kinase (ρι_3 kinase) Response to the signal. In most human solid tumors, such as breast cancer, prostate cancer, non-small cell lung cancer, gastrointestinal cancer, ovarian cancer, pancreatic cancer, etc., there are mutations and abnormal activation or overexpression of EGFR and/or Her-2. Thus, its association with tumor development has been confirmed. The platelet-derived growth factor receptor (PDGFR) and c-Kit of the RTKs Class III subfamily, similar to the ErbB family, are also activated by the dimerization process to transmit signals. This family member is closely related to the differentiation, proliferation and angiogenesis of tumor cells. For example, in small cell bronchial carcinoma, melanoma, breast cancer, and neuroblastoma, c_Kit is highly expressed or canine (see Schii tte et al., innovartis 3/2001). Especially in the gut glioma (GIST), gene mutations can continuously activate the c_Kit receptor, which leads to an increase in cell division rate, which in turn leads to genomic instability and canceration. (See weber et al, 乂 ^c〇7·22(14S), 9642 (2004)) Another important member of RTKs is the vascular endothelial growth factor receptor (VEGFR). VEGFR is directly involved in the process of angiogenesis, which induces endothelial cell proliferation and migration of ''''''''''''''''''''''' VEGF promotes tumor growth directly in tumor cells by the pr〇survivai mechanism. By research, it has been found that a large number of malignant solid tumors, such as lung cancer, breast cancer, ovarian cancer, pancreatic cancer and melanoma, can inhibit tumor growth by inhibiting VEGFR activity. Great application value. In addition, hepatocyte growth factor receptor c-Met (HGFR), a member of the RTKs family, has been shown to be closely related to tumorigenesis, invasion and metastasis, and cell motility enhancement (see Ma, P. C, etc.) 〇〇3b) 22,309-25; Maulik, G. et al. (2002b). 〇^〇々/this is (10)仏13,41_59). The main feature of tumor cells is the loss of control of genomic damage and signal regulatory pathways. Genomic damage leads to the alteration or loss of the biological function of some key regulatory proteins, which in turn destroys the signaling process. The abnormal production pathway enables tumor cells to survive and proliferate in the state of gene damage. As the basis for achieving these regulatory processes, PTKs are closely related to the occurrence and development of tumors, and thus become important targets for tumor therapy. It is hoped that by inhibiting one or more of the RTKs, physiological disturbances caused by abnormal proliferation of cells mediated by RTKs can be effectively improved and treated. A novel class of π-p and n-hybrid derivatives and their use as protein kinase inhibitors are disclosed in WO 2008/138232, wherein the disclosed example 53 is (疋^)-2 represented by formula (I) -(5-fluoro-2-oxooxy-1,2, diazepazine-3-indolyl)-5-(2-hydroxy-3-morpholin-4-yl-propyl)-3~ Aiwu 95166 201228664 -5, 6, 7, 8-tetrahydro-1 and -pyrrolo[3,2-c]azepine _4-ketone.

本申請人發現式（I)化合物在常規溶劑中溶解性差，不利於被製備成藥用劑型’並且降低了化合物的體内生物利用度，因此迫切需要開發式（I)化合物的新形式，解決這類化合物溶解性差’藥代吸收差的問題，以適合於常規製劑工藝。本發明的目的是提供式（I)化合物的可藥用的鹽形式，從而改善其物理化學性質和藥代動力學（Pharmac〇kinet ics) 特徵。【發明内容】本發明涉及式（I)化合物的可藥用的鹽，以及製備該鹽的方法。優選地’式（I)化合物的馬來酸鹽相對其他鹽和式（I)化合物本身在溶解度以及生物利用度和藥代動力學方面的優勢。The Applicant has found that the compound of the formula (I) has poor solubility in a conventional solvent, which is disadvantageous for being prepared into a pharmaceutical dosage form' and reduces the in vivo bioavailability of the compound, so that it is urgent to develop a new form of the compound of the formula (I) to solve Such compounds have poor solubility and poor drug absorption problems to suit the conventional formulation process. It is an object of the present invention to provide pharmaceutically acceptable salt forms of the compounds of formula (I), thereby improving their physicochemical properties and pharmacokinetic properties. SUMMARY OF THE INVENTION The present invention is directed to pharmaceutically acceptable salts of the compounds of formula (I), and to methods of preparing the salts. Preferably, the maleate salt of the compound of formula (I) has advantages over other salts and the compound of formula (I) in terms of solubility and bioavailability and pharmacokinetics.

本發明第一方面涉及式（I)所示的（足氟-2-側氧基-1，2-二氫-吲哚-3-次曱基）-5-(2-羥基-3-嗎啉-4- 7 95166 201228664 ^丙基>”基〜5,6,7，“氫_ 因-4-酮的可藥用㈧d 吖庚者有機鹽，it-步^縣本領域常規的無機鹽或趟、炉㈣，述的無機鹽選自鹽酸鹽、氫溴酸鹽選自？磺酸越…… ，所述的有機鹽、三氣醋酸二：=、酒石酸鹽，酸鹽、醋酸酸鹽、苯甲酸二；=、㈣酸鹽、枸櫞酸鹽、苯確萘嶒酸鹽、乳酸鹽或蘋果酸鹽，優選蒱 =鹽相：鹽、甲_馬來酸鹽。尤其是其馬來酸 …、+八他鹽和式（1)化合物本身在溶解度以及生物利用度和藥代動力學方面更具優勢。本發明第二方面涉及（^-2-(5-11-2-側氧基-u— 二氫-吲哚一次甲基）-5-(2-羥基-3-嗎啉-4-基-丙基i_3 一 :基-5, 6, 7, 8-四氫-if吡咯並[3, 2_c]吖庚因_4_酮的可藥用的鹽的製備方法，該化合物的製備可根據本領域常規的成鹽方法製備。具體而言，該方法包括將（亿幻_2_(5一氟一2一侧氧基一 1，2-二氫-吲哚-3-次曱基）-5-(2-羥基-3-嗎啉-4-基-丙基）一3—甲基一5, 6, 7, 8一四氫吡咯並[3, 2_c] β丫庚因-4-酮與相應的酸成鹽的步驟，其中該酸為選自磷酸、鹽酸、硫酸、硝酸、氫溴酸、甲磺酸、馬來酸、酒石酸、破轴酸、醋酸、三氟醋酸、富馬酸、檸:檬酸、枸櫞酸、苯續酸、苯曱酸、萘磺酸、乳酸或蘋果酸的無機酸或有機酸。本發明第三方面涉及一種藥物組合物，其含有治療有效劑量的（疋^)-2-(5-氟-2-侧氧基-1，2-二氫-吲哚-3-次 8 95166 201228664 曱基）-5-(2-經基—3一嗎琳+基一丙基）一3_甲基一5, 6, 了，8一四氫心吻各並[3,2一小丫庚因一4_嗎可藥用的载體。本發明的第四方面涉及（W_2_(5_l2_侧氧基 :2:二氫，3_次甲基)地射嗎琳—4—基-丙基）-3二基-5, 6, 7’ 8_四氫—…比嘻並[3, 2_小丫庚因_4一 ==樂用的鹽或其藥物組合物在製備治療與蛋白質激酶 $關的疾病的藥物中的用途。其中所述與蛋白質激酶有關的疾病選自與V刪-2, EGFR，職_2, ρ_，c_Kit， Γ!Γ，4FGFR相關的疾病，其中所述的疾病為癌症，選自肺癌、乳腺癌、表皮鱗癌或胃癌。本發明的第五方面涉及—種治療與蛋白質激酶有關該方法包括給予需要治療的患者有效治療 =的⑽2_n朴次甲 =5-(2-歸-3—嗎琳_4务丙基）_3m 6, 了，8_四 1合1吻各並[3’2_小丫庚因—4—酮的可藥用的鹽及其藥物本發明的第六方面涉及u，wn2_側氧基其;;二氫，朵—3-次甲基)-5-(2,基土 -甲基-5, 6, 7, 8-四氫督鱗並[3, 2_小丫庚因+ ==的鹽或其藥物組合物在製備蛋白質激酶抑制劑 Η的樂物中的用途，其中所述的蛋白激酶選自臟_2，腿， 2，PDGFR，c-Kit，c-Met 或 FGFR。本發明的第七方面涉及un(5、氟_2_側氧基 95166 9 201228664 二氫-啊-3_次甲基）基）-3-曱基_5, 6, 7, 8H Y馬^4~基~两鋼的可藥用的鹽或其藥物組合物療關的疾病的藥物，其中所述〜、/α%〜蛋白質激峰有腺癌、表皮鱗癌或胃癌。、、病為癌症，選自肺癌、乳經試驗比較，式（I)化入从& 生物度和藥誠軸力;來^在溶解度以及合物本身。勒力干方面優於其他鹽和式⑴化本發明關鍵原料式⑴化合物的合成方法式⑴解峨⑽(512_魏基_12_二氣一弓丨 W3-次曱基)-5-(2-羥基i嗎啉一4_基_丙基甲基 -5,6,7’8-四氫，-吡咯並[3,2_+丫庚因_4_酮的合：法根據聊08/138232中公開的實施例53所述的方法製備，因此將該公開内容作為參考文獻。【實施方式】以下結合實施例用於進一步描述本發明，但這些實施例並非限制著本發明的範圍。實施例化合物的結構是藉由核磁共振（⑴或/和質譜（MS) 來確疋的。NMR位移（δ)以百萬分之一（ppm)為單位。nmr 的測定是用Bruker AVANCE-400核磁儀，測定溶劑為氘代一甲基亞砜（d-DMSO) ’内標為四甲基矽烷（TMS)，化學位移疋以10 6(ppm)為單位。的測定用FINNIGAN LCQAd (ESI)質譜儀（生產商： 10 95166 201228664The first aspect of the invention relates to (Fluoro-2-oxo-1,2-dihydro-indol-3-indolyl)-5-(2-hydroxy-3-) represented by formula (I) Porphyrin-4- 7 95166 201228664 ^propyl >" base ~ 5,6,7, "hydrogen _ -4- ketone medicinal (eight) d 吖者 organic salt, it-step ^ county conventional inorganic in the field Salt or bismuth, furnace (four), the inorganic salt is selected from the group consisting of hydrochloride, hydrobromide is selected from the group consisting of sulfonic acid, the organic salt, tri-acetic acid two: =, tartrate, acid salt, acetic acid Acid salt, benzoic acid di;=, (tetra) acid salt, decanoate salt, benzoate naphthoate, lactate or malate, preferably 蒱=salt phase: salt, methyl-maleate, especially The maleic acid ..., the + octa salt and the compound of the formula (1) are themselves more advantageous in terms of solubility and bioavailability and pharmacokinetics. The second aspect of the invention relates to (^-2-(5-11-2-) Side oxy-u-dihydro-indole primary methyl)-5-(2-hydroxy-3-morpholin-4-yl-propyl i_3 one: yl-5, 6, 7, 8-tetrahydro- A process for the preparation of a pharmaceutically acceptable salt of pyrrolo[3,2_c]azepine-4-ketone, which can be prepared according to conventional salt formation in the art In particular, the method comprises the steps of: (Ethyl phthalate 2-(5-fluoro-2-one-oxy-1,2-dihydro-indole-3-indenyl)-5-(2-hydroxyl) -3-morpholin-4-yl-propyl)-3-methyl-5,6,7-8-tetrahydropyrrolo[3,2_c]β丫heptan-4-one is formed with the corresponding acid a step wherein the acid is selected from the group consisting of phosphoric acid, hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, methanesulfonic acid, maleic acid, tartaric acid, arachidonic acid, acetic acid, trifluoroacetic acid, fumaric acid, and citric acid. An inorganic or organic acid of citric acid, benzoic acid, benzoic acid, naphthalenesulfonic acid, lactic acid or malic acid. A third aspect of the invention relates to a pharmaceutical composition comprising a therapeutically effective dose of (疋^)-2 -(5-fluoro-2-oxooxy-1,2-dihydro-indole-3-times 8 95166 201228664 fluorenyl)-5-(2-pyridyl-3 merinyl-yl-propyl) A 3-methyl-5,6,8-tetrahydro-nuclear kiss and a pharmaceutically acceptable carrier. The fourth aspect of the invention relates to (W_2_( 5_l2_sideoxy: 2: dihydrogen, 3_methine) shots 吗琳-4-yl-propyl)-3diyl-5, 6, 7' 8_tetrahydro-... 3, 2_小丫庚_4一==The use of a salt or a pharmaceutical composition thereof for the preparation of a medicament for treating a disease associated with a protein kinase, wherein the protein kinase-related disease is selected from the group consisting of V deleted-2, EGFR, _ 2, ρ_, c_Kit, Γ!Γ, 4FGFR-related diseases, wherein the disease is cancer, selected from the group consisting of lung cancer, breast cancer, epidermal squamous cell carcinoma or gastric cancer. A fifth aspect of the invention relates to a treatment associated with a protein kinase, the method comprising administering to a patient in need of treatment an effective treatment = (10) 2 _ n 朴甲 = 5 - (2- -3 - 3 - 琳 _ 4 propyl) _ 3 m 6 , a pharmaceutically acceptable salt of 8_4 1 in 1 kiss each [3'2_small indenyl-4-ketone and a medicament thereof. The sixth aspect of the invention relates to u, wn2_sideoxy; Dihydrogen, flower-3-methylidene-5-(2, basal-methyl-5, 6, 7, 8-tetrahydro-scaled [3, 2_small 丫geng + == Use of a salt or a pharmaceutical composition thereof for the preparation of a protein kinase inhibitor, wherein the protein kinase is selected from the group consisting of viscera-2, leg, 2, PDGFR, c-Kit, c-Met or FGFR. The seventh aspect relates to un(5, fluoro_2_sideoxy 95166 9 201228664 dihydro-ah-3_methine) benzyl-3-yl _5, 6, 7, 8H Y horse ^4~ A drug for treating a disease of a pharmaceutically acceptable salt or a pharmaceutical composition thereof, wherein the ~, /α%~ protein peak has adenocarcinoma, epidermal squamous cell carcinoma or gastric cancer. The disease is cancer, which is selected from the group consisting of lung cancer and milk-to-milk test. Formula (I) is converted into & bioavailability and drug affinity; and in solubility and composition itself. The method of synthesizing Leligan is superior to other salts and formula (1) The key material of the invention (1) is synthesized by the formula (1). (10) (512_Weiji_12_二气一弓丨W3-次曱基)-5-( 2-hydroxyimorpholine-4-yl-propylmethyl-5,6,7'8-tetrahydro,-pyrrolo[3,2_+azepine_4_one combination: according to Liao 08/ The method described in Example 53 of the disclosure of 138, 232, the disclosure of which is hereby incorporated by reference. The structure of the compound is confirmed by nuclear magnetic resonance ((1) or / and mass spectrometry (MS). The NMR shift (δ) is in parts per million (ppm). The nmr is measured using Bruker AVANCE-400 NMR. The solvent was determined to be deuterated monomethyl sulfoxide (d-DMSO). The internal standard was tetramethyl decane (TMS) and the chemical shift 疋 was in units of 10 6 (ppm). The measurement was performed using FINNIGAN LCQAd (ESI) mass spectrometry. Instrument (manufacturer: 10 95166 201228664

Thermo,型號：Finnigan LCQ advantage MAX)。 HPLC的測定使用安捷倫1200DAD高壓液相色譜儀 (Sunfire C18 150x4. 6mm 色譜管柱）和 Waters 2695-2996 高壓液相色譜儀（Gimini C18 150x4. 6mm色譜柱）。管柱色譜法一般使用煙臺黃海矽膠200〜300目矽膠為載體。本發明的起始原料是已知的，並且可以在市場上購買到’購買自 ABCR GmbH & Co. KG，Acros Organics，Aldrich Chemical Company等公司’或者可以採用或者按照本領域已知的方法來合成。實施例中無特殊說明，反應均在氬氣氛或氮氣氛下進行。氬氣氛或氮氣氛是指反應瓶連接一個約1L容積的氬氣或氮氣氣球。氫氣氛是指反應瓶連接一個約1L容積的氫氣氣球。實施例中無特殊說明，溶液是指水溶液。實施例中無特殊說明，反應的溫度為室溫。室溫為最適宜的反應溫度，為2〇。〇至3〇°C。實施例中的反應進程的監測採用薄層色譜法（TLC)，反應所使用的展開劑的體系有：二氯曱烷和曱醇體系，正己烷和乙酸乙酯體系，石油醚和乙酸乙酯體系，丙酮，溶劑的體積比根據化合物的極性不同而進行調節。管柱層析的洗脫劑的體系包括：A:二氣曱烧、曱醇和丙酮體系，B:正己烷和乙酸乙酯體系，溶劑的體積比根 95166 11 201228664 據化合物的極性不同而進行調節，也可以加入少量的氣水和醋酸等進行調節。實施例1 (yp，^)-2-(5-氟-2-側氧基-1，2-二氬-吲哚-3-次曱基） -5_(2-羥基-3-嗎啉-4-基-丙基）-3-曱基-5, 6, 7, 8-四氫 -1#-吡咯並[3, 2-c]吖庚因-4-酮馬來酸鹽Thermo, model: Finnigan LCQ advantage MAX). The HPLC was measured using an Agilent 1200 DAD high pressure liquid chromatograph (Sunfire C18 150 x 4. 6 mm chromatography column) and a Waters 2695-2996 high pressure liquid chromatograph (Gimini C18 150 x 4. 6 mm column). Pipe column chromatography generally uses Yantai Huanghai Silicone 200~300 mesh gelatin as a carrier. The starting materials of the present invention are known and commercially available from 'ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, etc.' or may be employed or according to methods known in the art. synthesis. Unless otherwise stated in the examples, the reactions were all carried out under an argon atmosphere or a nitrogen atmosphere. An argon atmosphere or a nitrogen atmosphere means that the reaction flask is connected to an argon or nitrogen balloon having a volume of about 1 L. The hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon of about 1 L volume. Unless otherwise stated in the examples, the solution means an aqueous solution. There is no particular description in the examples, and the reaction temperature is room temperature. The optimum reaction temperature at room temperature is 2 Torr. 〇 to 3〇 °C. The progress of the reaction in the examples was monitored by thin layer chromatography (TLC). The systems used for the reaction were: dichlorosilane and decyl alcohol systems, n-hexane and ethyl acetate systems, petroleum ether and ethyl acetate. The volume ratio of the system, acetone, and solvent is adjusted depending on the polarity of the compound. The system of the eluent for column chromatography includes: A: dioxane, decyl alcohol and acetone system, B: n-hexane and ethyl acetate system, the volume ratio of the solvent is 95166 11 201228664, which is adjusted according to the polarity of the compound. It can also be adjusted by adding a small amount of gas water and acetic acid. Example 1 (yp,^)-2-(5-fluoro-2-oxooxy-1,2-diar-indol-3-indolyl)-5-(2-hydroxy-3-morpholine- 4-yl-propyl)-3-indolyl-5, 6, 7, 8-tetrahydro-1#-pyrrolo[3,2-c]azepine-4-one maleate

第一步 5-曱醯基-3-甲基-1及-吡咯-2,4-二羧酸2-叔丁酯4-乙酯將3, 5-二曱基-1及-吡咯-2, 4-二羧酸2-叔丁酯4-乙酯la(30 g，0· 11 mol)授拌下溶解於300 mL四氫吱喃中，加入3 6 0 mL醋酸和3 0 0 mL水。授拌均勻，一次性加入硝 12 95166 201228664 酸鈽胺（246 g，0. 45 mol)。攪拌反應0. 5小時。將反應液倒入800 mL冰水中’攪拌〇. 5小時，過濾，固體真空乾燥，得到標題標題產物5-曱醯基-3-甲基-1及-吡咯-2, 4-二竣酸2-叔丁酯4-乙酯lb(31. 13 g，淺黃色固體），產率： 98%。 MS ra/z (ESI): 282.0[M+1] 第二步 5_(2_乙氧k基-乙稀基）_3_曱基-1及-〇比略-2，4_二緩酸2- 叔丁酯4-乙酯將5-甲醯基-3-甲基-1)7-吼洛-2, 4-二羧酸2-叔丁酯 4- 乙酯lb(23 g ’ 81· 7 mmol)和（乙酯基亞曱基）三苯基正膦（34. 66 g’ 99. 4 mmol)攪拌下溶解於450 mL四氫呋喃中，攪拌反應12小時。反應液減壓濃縮，用矽膠柱色譜法以洗脫劑體系B純化所得殘餘物，得到標題產物5-(2-乙氧羰基-乙烯基）-3-曱基-1及-吡咯-2, 4-二羧酸2-叔丁酯4-乙酯lc(24 g，淺黃色固體），產率：84%。 MS m/z (ESI): 352.1[M+1] 第三步 5- （2-乙氧獄基-乙基）-3 -曱基_1及-0比嘻-2, 4-二叛酸2-叔丁酯4-乙酯氫氣氛下，將5-(2-乙氧羰基-乙烯基）-3-甲基-1及-吡咯-2, 4-二羥酸 2-叔丁酯 4-乙酯 lc(24 g，68. 3 mmol) 攪拌下溶解於180 inL無水乙醇中，加入2. 44 g 10%鈀/碳，攪拌反應12小時。過濾，濾餅用少量乙醇洗滌，收集濾液’ 13 95166 201228664 減壓濃縮，得到標題產物5-(2-乙氧羰基-乙基）—3-甲基 -1及比嘻-2, 4-二叛酸2-叔丁酯4-乙酯Id (23 g，白色固體），產率：95%。 MS m/z (ESI): 354.4[M+1] 第四步 5-(2-羧基-乙基）-3-曱基-IF吡咯-2, 4-二羧酸2-叔丁酯 4-乙酯將5-(2-乙氧幾基-乙基）-3-甲基-1及_0比17各_2,4-二幾_ 酸2-叔丁酯4-乙酯ld(23. 6 g，66· 8 mmol)攪拌下溶解於190 mL四氫。夫喃和90 mL曱醇中，滴加80 mL 10 Μ氫氧化鋰溶液，攪拌反應1小時。反應液減壓濃縮。冰浴下，滴加2 Μ鹽酸至反應液pH為2。過濾，真空乾燥後得到標題產物5-(2-羧基-乙基）-3-曱基-1及-吡咯-2, 4-二羧酸 2-叔丁酯4-乙酯le(24 g，白色固體），產率：98%。 MS m/z (ESI)： 326.1[M+1] 第五步 5-(3-羥基-丙基）-3-甲基-IF吡咯-2, 4-二羧酸2-叔丁酯 4-乙酯將5-(2-羧基-乙基）-3-曱基-1及-吡咯-2, 4-二羧酸 2-叔丁酯4-乙酯le(9. 75 g，30 mmol)授拌下溶解於90 mL 無水四氫0夫喃中。在-10至-5°C下，緩慢滴加90 mL 1 Μ 硼烷的四氫呋喃溶液。室溫下攪拌反應2至3小時。反應液減壓濃縮，加入1 〇〇 mL飽和碳酸氫納溶液和1 〇〇 mL乙酸乙酯’用乙酸乙酯萃取（100 mLx3)，合併有機相，用飽 95166 14 201228664 和食鹽水洗滌（100 mL) ’無水硫酸鎂乾燥，過濾，滤液濃縮得到標題產物5-(3-羥基-丙基）—3-甲基-Ιθ-α比σ各_2, 4-二羧酸2-叔丁酯4-乙酯If (9. 2 g，淺黃色油狀物），產率：98% 。 MS m/z (ESI): 312.3[M+1] 第六步 5-(3 -曱石黃酿氧基-丙基）-3 -曱基及-〇比p各一2, 4-二缓酸2_ 叔丁酯4-乙酯The first step is 5-mercapto-3-methyl-1 and -pyrrole-2,4-dicarboxylic acid 2-tert-butyl ester 4-ethyl ester 3, 5-dimercapto-1 and -pyrrole-2 , 4-tert-butyl 4-carboxylate la (30 g, 0·11 mol) was dissolved in 300 mL of tetrahydrofuran, and added to 660 mL of acetic acid and 300 mL of water. . Uniform mixing, one-time addition of nitrate 12 95166 201228664 decylamine (246 g, 0. 45 mol). 5小时。 The reaction was stirred for 0.5 hours. The reaction solution was poured into 800 mL of ice water and stirred for 5 hours, filtered, and dried in vacuo to give the title product 5-mercapto-3-methyl-1 and -pyrrole-2,4-didecanoic acid 2 - tert-Butyl 4-ethyl ester lb (31.13 g, pale yellow solid), yield: 98%. MS ra/z (ESI): 282.0 [M+1] The second step 5_(2_ ethoxyk-ethylidene)_3_mercapto-1 and - oxime-2,4_di-acid 2 - tert-Butyl 4-ethyl ester 5-butyryl-3-methyl-1)7-indolo-2,4-dicarboxylic acid 2-tert-butyl ester 4-ethyl ester lb (23 g '81· 7 mmol) and (ethyl ester sulfhydryl) triphenylphosphorane (34.66 g '99. 4 mmol) were dissolved in 450 mL of tetrahydrofuran with stirring, and the reaction was stirred for 12 hours. The reaction mixture was concentrated under reduced pressure. EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj 4-Dicarboxylic acid 2-tert-butyl ester 4-ethyl ester lc (24 g, pale yellow solid), yield: 84%. MS m/z (ESI): 352.1 [M+1] Step 3 5-(2-Ethoxyphenyl-ethyl)-3-indolyl-1 and -0-嘻-2,4-di-oroxic acid 2-(2-ethoxycarbonyl-vinyl)-3-methyl-1 and-pyrrole-2,4-dihydroxy acid 2-tert-butyl ester 4 under hydrogen atmosphere of 2-tert-butyl ester 4-ethyl ester Ethyl ester lc (24 g, 68.3 mmol) was dissolved in 180 inL absolute ethanol with stirring, 2.44 g of 10% palladium on carbon was added, and the reaction was stirred for 12 hours. Filtration, the filter cake was washed with a small amount of ethanol, and the filtrate was collected. <RTI ID=0.0>&&&&&&&&&&&&&&&&&&&&& Retinoic acid 2-tert-butyl ester 4-ethyl ester Id (23 g, white solid), yield: 95%. MS m/z (ESI): 354.4 [M + 1] Step 4 5-(2-carboxy-ethyl)-3-indolyl-IFpyrrole-2, 4-dicarboxylic acid 2-tert-butyl ester 4- Ethyl ester 5-(2-ethoxyxo-ethyl)-3-methyl-1 and _0 to 17 each 2,2-di-acid 2-tert-butyl ester 4-ethyl ester ld (23 6 g, 66·8 mmol) was dissolved in 190 mL of tetrahydrogen under stirring. 80 mL of 10 Μ lithium hydride solution was added dropwise to the sulphur and 90 mL of sterol, and the reaction was stirred for 1 hour. The reaction solution was concentrated under reduced pressure. Under ice bath, 2 Μ hydrochloric acid was added dropwise until the pH of the reaction solution was 2. Filtration and drying in vacuo gave the title product 5-(2-carboxy-ethyl)-3-indolyl-1 and -pyrrole-2, 4-dicarboxylic acid 2-tert-butyl ester 4-ethyl ester (24 g, White solid), Yield: 98%. MS m/z (ESI): 326.1 [M + 1] Step 5 5-(3-hydroxy-propyl)-3-methyl-IFpyrrole-2, 4-dicarboxylic acid 2-tert-butyl ester 4- Ethyl ester 5-(2-carboxy-ethyl)-3-indolyl-1 and 2-pyrryl-2,4-dicarboxylic acid 2-tert-butyl ester 4-ethyl ester le (9. 75 g, 30 mmol) Dissolved in 90 mL of anhydrous tetrahydrofuran. 90 mL of a solution of 1 Torr borane in tetrahydrofuran was slowly added dropwise at -10 to -5 °C. The reaction was stirred at room temperature for 2 to 3 hours. The reaction mixture was concentrated under reduced pressure. EtOAc EtOAc (EtOAc)EtOAc. Drying with anhydrous magnesium sulfate, filtration, and concentration of the filtrate to give the title product 5-(3-hydroxy-propyl)- 3-methyl- Ι θ-α ratio σ _2, 4-dicarboxylic acid 2-tert-butyl ester 4 - ethyl ester If (9.2 g, pale yellow oil), yield: 98%. MS m/z (ESI): 312.3 [M+1] Step 6 5-(3 - vermiculite yellow-oxyl-propyl)-3 -fluorenyl and -fluorene ratio p 2, 4- 2 Acid 2_ tert-butyl ester 4-ethyl ester

將5-(3-羥基-丙基）-3-甲基-1及-吡咯-2, 4-二羧酸 2-叔丁酯4-乙酯If (9. 20 g，30 mmol)攪拌下溶解於150 mL 二氣曱烷中。在-10°C下，依次滴加三乙胺（7 mL，50 mmol) 和甲磺醯氯（3. 5 mL，45 mmol)。室溫攪拌反應4小時。加入少量冰水，依次用0.5 Μ鹽酸（80 mLx2)、飽和碳酸鈉溶液（80 mLx2)、飽和食鹽水洗滌（8〇 mL)，用無水硫酸鎂乾燥，過濾、，濾、液減壓濃縮，得到標題產物5-(3-曱續醯氧基-丙基）-3-曱基-1万-吡咯-2, 4-二羧酸2-叔丁酯4-乙酯 lg(11.4 g，棕色油狀物），產率：99%。 MS m/z (ESI)： 390.5[M+1] 第七步 (友)-4-環氧乙烷基曱基-嗎啉將嗎啉lh(8. 7 mL，0. 1 mol)攪拌下溶解於4. 5 mL叔丁醇中，冰浴下，滴加（皮)-(-)-環氧氣丙烷（8.丨mL，〇.丄 m〇l)，室溫攪拌24小時。10°C下，滴加6〇此1· 67 M叔丁醇鉀的四氫咬喃溶液，攪拌反應30分鐘。反應液減壓 95166 15 201228664 濃縮，殘留物中加入50 mL水，用二氣曱烷萃取（1〇〇 mLx2)，合併有機相，用飽和食鹽水溶液洗滌（100 mL)，無水硫酸鎮乾燥’過滤’慮液濃縮得到標題產物（及)-4-環氧乙烧基曱基-嗎琳li(12.7 g’黃色油狀物），產率：88. 8%。 MS m/z (ESI): 144.4[M+1] 第八步 (5〇_l-胺基-3-嗎琳-4-基-異丙醇冰浴下，將（Λ〇-4-環氧乙烧基曱基-嗎琳1 i(6. 3 g， 44 mmol)緩慢滴入450 mL 25%氨水中，室溫攪拌反應is 小時。反應液減壓濃縮’得到標題產物（5") -1 -胺基-3 -嗎琳 -4-基-異丙醇lj(7 g，白色固體），產率：99%。 MS m/z (ESI): 161.1[M+l] 第九步 (5〇-5-[3-( 2-經基-3-嗎淋-4-基-丙胺基）-丙基]-3-曱基 -1及~吡咯-2, 4-二羥酸2-叔丁酯4-乙酯將5-(3-曱磺醯氧基-丙基）-3-曱基-1及-吡咯-2, 4-二羧酸2-叔丁酯4-乙酯lg(l· 13 g，2. 9 mmol)攪拌下溶解於5. 6 inL —氣甲院中，加入（iS)-!·-胺基-3_嗎琳-4-基-異丙醇1 j(0. 93 g，5. 8 mmol)。授拌反應12小時，繼續於 45°C下加熱反應14小時。反應液中加入15 mL飽和食鹽水溶液，用二氣甲烷萃取（20 mLx3)，合併有機相，減壓濃縮，用矽膠柱色譜法以洗脫劑體系A純化所得殘餘物，得到標題產物（Λ-5-[3-(2-羥基-3-嗎啉-4-基-丙胺基）-丙基] -3-曱基-1万-吡咯-2, 4-二羥酸2-叔丁酯4-乙酯lk(600 16 95166 201228664 mg，無色油狀物），產率：72. 5%。 MS m/z (ESI): 454.2[M+1] 第十步 U)-5-(2-羥基-3-嗎啉-4-基-丙基）-3-曱基_5, 6, 7, 8_四氫-1及-吡咯並[3, 2-c]吖庚因-4-酮將（5)-5-[3-(2-羥基-3-嗎啉-4-基-丙胺基）-丙基] -3-甲基-1及-吡咯-2,4-二羥酸2-叔丁酯4-乙酯比（580 mg ’ 1. 28腿〇1)攪拌下溶解於6 mL曱苯中。冰浴下，滴加 1. 9 mL 2 Μ二甲基鋁的曱苯溶液。回流反應24小時。反應液減壓濃縮，加入20 mL 6 Μ鹽酸，攪拌20分鐘。冰浴下，滴加12Μ虱氧化鈉溶液至反應液pH為12，用二氣曱烷萃取（50 mLx2)，合併有機相，減壓濃縮，用矽膠柱色譜法以洗脫劑體系A純化所得殘餘物，得到標題產物（妁 (2-羥基-3-嗎啉-4-基-丙基）_3_曱基_5, 6, 7, 8一四氫_liy_ 吡咯並[3, 2-c]吖庚因-4-酮im(3〇〇呢，白色固體），產率： 57. 6%。 MS ra/z (ESI)： 308.2[M+1] 第十一步 (及)-5-(2-羥基-3-嗎啉—4-基-丙基）_3_曱基_4-侧氧基一1’4’ 5’ 6, 7, 8-六氫-吡咯並[3, 2_c]吖庚因_2_甲醛將氣亞甲基二曱基氣化胺（130 mg，0. 98 mmol)攪拌下溶解於3 mL二氯甲烷中。〇。(：下，加入2 mL (i?)-5-(2-經基-3-嗎嘛-4~基-丙基）_3_甲基_5, 6, 7, 8—四氫_1)7_吡咯並[3, 2 c] 丫庚因-4-嗣 im(3〇〇 mg，〇 98 mmol)的二氯甲 17 95166 201228664 院溶液。室溫攪;摔反應2 0分鐘。依次加入1 〇 mL 12 Μ氫氧化鈉溶液和10 mL飽和食鹽水溶液，用二氣曱烧和甲醇的混合溶劑（V:V=10:1)萃取（100 mLx3)，合併有機相，用飽和食鹽水溶液洗滌（100 mL)，無水硫酸鎂乾燥，過渡，濾液減壓濃縮，用矽膠柱色譜法以洗脫劑體系A純化所得殘餘物’得到標題產物（i?)-5-(2-羥基-3-嗎啉-4-基-丙基）-3-甲基-4-侧氧基-1，4, 5, 6, 7, 8-六氫比洛並[3, 2-c] 吖庚因-2-曱醛ln(200 mg，白色固體），產率：610/〇。 MS m/z (ESI)： 336.2[M+1] 第十二步 (疋^)-2-(5-氟-2-側氧基-1，2-二氫-吲哚-3-次甲基）-5-(2-羥基-3-嗎啉-4-基-丙基）-3-甲基-5, 6, 7, 8、四氫-1及比洛並[3, 2-c]吖庚因-4-酮將（及)-5-（2-經基-3-嗎琳-4-基-丙基）-3_曱基-4-側氧基-1，4, 5, 6, 7, 8-六氫比口各並[3, 2-c]吖庚因-2-曱駿 ln(50 mg，0. 15 mmol)攪拌下溶解於261//L乙醇中，加入 5-氟-1’3-二氫-α引d朵-2-酮（20 mg，0. 13 mmol)和呢交 (7. 3 eL，0. 074 mmol)。80〇C下，避光攪拌反應2小時。冷卻至室溫，過濾’真空乾燥得到標題產物（尤幻_2~(队氟-2-側氧基-1，2-二氫-吲哚-3-次曱基）-5-(2-羥基-3、嗎啉-4-基-丙基）-3-甲基-5, 6, 7, 8-四氫-1#-吡咯並[3, 吖庚因-4-酮lp (40 mg，黃色固體），產率：57%。 MS m/z (ESI): 469.2[M+1] iNMRMOOMHz，i/-DMS0，ppm): 513.73(s，1H)，10.9i(s 18 95166 201228664 lH)，7.76〜7.78(m，lH)，7.75(s，1H)，6.91〜6.94(m，iH) 6.84〜6.87(m’lH)，4.72〜4.73(d，lH)，3.9〇(m，lH) ’5-(3-Hydroxy-propyl)-3-methyl-1 and-pyrrole-2,4-dicarboxylic acid 2-tert-butyl ester 4-ethyl ester If (9. 20 g, 30 mmol) was stirred Dissolved in 150 mL of dioxane. Triethylamine (7 mL, 50 mmol) and methanesulfonium chloride (3.5 mL, 45 mmol) were added dropwise at -10 °C. The reaction was stirred at room temperature for 4 hours. The mixture was washed with aq. The title product 5-(3-indoleoxy-propyl)-3-indolyl-1 --pyrrole-2,4-dicarboxylic acid 2-tert-butyl ester 4-ethyl ester lg (11.4 g, brown Oil), yield: 99%. MS m/z (ESI): 390.5 [M+1] Step 7 (Friend)-4-Ethylene oxide-mercapto-morpholine with morpholine lh (8.7 mL, 0.1 mol) The solution was dissolved in 4.5 mL of tert-butanol, and (p)-(-)-epoxypropane (8. 丨mL, 〇.丄m〇l) was added dropwise under ice-cooling, and stirred at room temperature for 24 hours. At 10 ° C, 6 〇 of a tetrahydrogenated solution of 1.67 M potassium t-butoxide was added dropwise, and the reaction was stirred for 30 minutes. The reaction solution was decompressed with 95166 15 201228664, and the residue was added with 50 mL of water, and the mixture was extracted with dioxane (1 〇〇mL×2). The organic phase was combined and washed with saturated aqueous sodium chloride (100 mL) Concentration of the title product (and) 4-epoxyethyl decyl-m-lin-li (12.7 g (yellow oil)), yield: 88.8%. MS m/z (ESI): 144.4 [M+1] Step 8 (5〇_l-amino-3-morphin-4-yl-isopropanol under ice bath, (Λ〇-4-ring) Oxyethylidene-mercapto-Merlin 1 i (6.3 g, 44 mmol) was slowly dropped into 450 mL of 25% aqueous ammonia, and the reaction was stirred at room temperature for one hour. The reaction mixture was concentrated under reduced pressure to give the title product (5 ") -1 -Amino-3-merin-4-yl-isopropanol lj (7 g, white solid), yield: 99%. MS m/z (ESI): 161.1 [M+l] (5〇-5-[3-(2-Pyryl-3-ylide-4-yl-propylamino)-propyl]-3-indolyl-1 and ~pyrrole-2,4-dihydroxy acid 2 -tert-butyl ester 4-ethyl ester 5-(3-oxasulfonyloxy-propyl)-3-indolyl-1 and -pyrrole-2,4-dicarboxylic acid 2-tert-butyl ester 4-ethyl ester Lg(l·13 g, 2. 9 mmol) was dissolved in 5. 6 inL - gas institute, and (iS)-!--amino-3_morphin-4-yl-isopropanol 1 was added. j (0. 93 g, 5. 8 mmol). The reaction was stirred for 12 hours, and the reaction was further heated at 45 ° C for 14 hours. 15 mL of a saturated saline solution was added to the reaction mixture, and extracted with di-methane (20 mL×3). The combined organic layers were concentrated under reduced pressure and purified eluted elut elut Λ-5-[3-(2-hydroxy-3-morpholin-4-yl-propylamino)-propyl]-3-mercapto-1,000-pyrrole-2, 4-dihydroxy acid 2-tert-butyl Ester 4-ethyl ester lk (600 16 95166 201228664 mg, colorless oil), yield: 72. 5% MS m/z (ESI): 454.2 [M+1] Step 10 U)-5-( 2-hydroxy-3-morpholin-4-yl-propyl)-3-indenyl_5, 6, 7, 8-tetrahydro-1 and-pyrrolo[3,2-c]azepine-4 -ketone will be (5)-5-[3-(2-hydroxy-3-morpholin-4-yl-propylamino)-propyl]-3-methyl-1 and-pyrrole-2,4-dihydroxy The acid 2-tert-butyl ester 4-ethyl ester ratio (580 mg ' 1. 28 leg 〇 1) was dissolved in 6 mL of hydrazine under stirring. Under ice bath, 1. 9 mL of Μ dimethyl aluminum ruthenium was added dropwise. The benzene solution was refluxed for 24 hours. The reaction solution was concentrated under reduced pressure, and 20 mL of 6 hydrazine hydrochloric acid was added and stirred for 20 minutes. Under ice bath, 12 Μ虱 sodium oxide solution was added dropwise until the pH of the reaction mixture was 12, and extracted with dioxane ( 50 mL x 2), the combined organic phases were evaporated,jjjjjjjjjjjjj _3_曱基_5, 6, 7, 8-tetrahydro_liy_pyrrolo[3,2-c]azepine-4-one im (3〇〇, white solid ), Yield: 57.6%. MS ra/z (ESI): 308.2 [M+1] Step 11 (and)-5-(2-hydroxy-3-morpholine-4-yl-propyl)_3_indolyl_4-side oxygen Base 1'4' 5' 6, 7, 8-hexahydro-pyrrolo[3, 2_c]azepine _2_formaldehyde will be a gas methylene dimercapto gasified amine (130 mg, 0. 98 mmol Dissolved in 3 mL of dichloromethane with stirring. Hey. (:, add 2 mL (i?)-5-(2-pyridyl-3-??-4~yl-propyl)_3_methyl_5, 6, 7, 8-tetrahydro-1) 7_pyrrolo[3, 2 c] azepine-4-pyim (3〇〇mg, 〇98 mmol) of dichloromethyl 17 95166 201228664 Institute solution. Stir at room temperature; drop reaction for 20 minutes. Add 1 〇mL of 12 Μ sodium hydroxide solution and 10 mL of saturated saline solution, and extract (100 mL×3) with a mixture of two gas and methanol (V:V=10:1), and combine the organic phase with saturated salt. The title product (i?)-5-(2-hydroxy-) was obtained by washing with aqueous solution (100 mL), dried over anhydrous magnesium sulfate, and the filtrate was concentrated under reduced pressure. 3-morpholin-4-yl-propyl)-3-methyl-4-oxo-1,4,5, 6, 7, 8-hexahydropyrolo[3,2-c] Benzene furfural ln (200 mg, white solid), yield: 610 / 〇. MS m/z (ESI): 336.2 [M+1] Step 12 (疋^)-2-(5-fluoro-2-oxooxy-1,2-dihydro-indole-3-trimethyl 5-(2-hydroxy-3-morpholin-4-yl-propyl)-3-methyl-5, 6, 7, 8, tetrahydro-1 and biro[3, 2-c吖glycan-4-one ketone (and)-5-(2-yl-3-ylidene-4-yl-propyl)-3-mercapto-4-yloxy-1,4,5 , 6, 7, 8- hexahydrogen, each [3, 2-c] 吖hine-2- jun ln (50 mg, 0.15 mmol) dissolved in 261 / / L ethanol, added 5-Fluoro-1'3-dihydro-α-d-butan-2-one (20 mg, 0.13 mmol) and saponin (7.3 g, 0.074 mmol). The reaction was stirred for 2 hours in the dark at 80 °C. Cool to room temperature, filter and 'vacuum dry to give the title product (Ultra 2-1~(Tetrafluoro-2-epoxy-1,2-dihydro-indole-3-indenyl)-5-(2- Hydroxy-3,morpholin-4-yl-propyl)-3-methyl-5, 6, 7, 8-tetrahydro-1#-pyrrolo[3, azepin-4-one lp (40 mg , (yellow solid), Yield: 57%. MS m/z (ESI): 469.2[M+1] iNMRMOO MHz, i/-DMS0, ppm): 513.73 (s, 1H), 10.9i (s 18 95166 201228664 lH ), 7.76~7.78 (m, lH), 7.75 (s, 1H), 6.91~6.94 (m, iH) 6.84~6.87 (m'lH), 4.72~4.73 (d, lH), 3.9 〇 (m, lH) ) '

3.75〜3.79(dd，lH)，3.57〜3.59(t，4H)，3.38〜3.35Q3.75~3.79(dd,lH), 3.57~3.59(t,4H),3.38~3.35Q

2H)，3.14〜3.19(dd，lH)，2.92〜2.95(t，2H)，2.46(s，3H 2.42〜2.51〇!1，41〇,2.29至2.31(121〇,2.08(111，’2 第十三步 (尤7)-2-(5-氟-2-侧氧基-1，2-二氫-吲哚_3一次曱基） -5-(2-經基-3-嗎琳-4-基-丙基）-3-曱基_5, 6, 7, 8〜四氣 -1F吼洛並[3,2_c]n丫庚因酮馬來酸鹽 mm〇i) 將（及，幻-2-(5-氟-2-侧氧基-1，2-二氫—吲哚一3〜次甲基）-5-(2-經基-3-嗎琳-4-基-丙基）~3-曱基-5, 6, 7 8 氫各並[3, 2-c]吖庚因-4-酮 lp (731 mg，1. 56 和馬來酸（217 mg ’ 1· 87 mmol)授拌下溶解於τ ^ τ醇中，在40°C下攪拌反應20分鐘。過濾，濾液減壓濃縮，加入50 mL乙腈，回流反應20分鐘。冷卻至室溫，過廣得到（皮，^)-2-(5-氟-2-側氧基-1，2-二氫-吲哚_3-次曱基）-5-(2-羥基-3-嗎淋-4-基-丙基）-3-甲基-5, 6, 7, 8-四氫-1及-0比p各並[3, 2-c]吖庚因-4-酮馬來酸鹽1(831 mg，黃色固體），產率：91. 1%。 MS m/z (ESI): 469.2[M+1] WNMRUOOMHz，士DMSO，ppm): 5 13.76(s，1H)，l〇.93(s， lH)，7.77〜7_80(m，lH)，7.76(s，lH)，6.93〜6.98(m，lH)， 6.85〜6.88(m，lH)，6.05(s，2H)，4.19(d，lH)，3.63〜3.84 (m，4H),3.60〜3.61(m，lH)，3.43~3.46(m，4H)，3.31(m， 19 95166 201228664 2H)，3.13~3.18(m，3H)，2.96〜3.00(m，3H)，2.48(s，3H)， 2. 10〜2. 13(m，2H) 實施例2 (y?’幻-2-(5-氟-2-侧氧基-1，2-二氫-吲哚-3-次曱基） -5-(2-羥基-3-嗎啉-4-基-丙基）-3-曱基一5, 6, 7, 8-四氫 -1皮-吡咯並[3, 2-c]吖庚因-4-酮蘋果酸鹽2H), 3.14~3.19(dd,lH), 2.92~2.95(t,2H), 2.46(s,3H 2.42~2.51〇!1,41〇, 2.29 to 2.31 (121〇,2.08(111,'2 Thirteen Steps (Especial 7)-2-(5-Fluoro-2-Sideoxy-1,2-Dihydro-indole_3 Monodecyl) -5-(2-Ph--3-inline- 4-yl-propyl)-3-indenyl_5, 6, 7, 8~tetraki-1F吼洛[3,2_c]n丫heptanone maleate salt mm〇i) will (and, Magic-2-(5-fluoro-2-indolyl-1,2-dihydro-indole-3~methine)-5-(2-pyridyl-3-morphin-4-yl-propyl Base) ~3-mercapto-5, 6, 7 8 Hydrogen each [3, 2-c]azepin-4-one lp (731 mg, 1.56 and maleic acid (217 mg '1·87) Ment) Dissolved in τ ^ tau alcohol under stirring, stirred at 20 ° C for 20 minutes, filtered, concentrated under reduced pressure, added 50 mL of acetonitrile, refluxed for 20 minutes, cooled to room temperature, over-boiled ,^)-2-(5-fluoro-2-indolyl-1,2-dihydro-indole-3-indolyl)-5-(2-hydroxy-3-indol-4-yl- Propyl)-3-methyl-5, 6, 7, 8-tetrahydro-1 and-0 ratio p and [3, 2-c]azepin-4-one maleate 1 (831 mg , yellow solid), Yield: 91.1%. MS m/z (ESI): 469.2 [M+1] WNMRUOOMHz, DMSO Ppm): 5 13.76(s,1H), l〇.93(s, lH), 7.77~7_80(m,lH), 7.76(s,lH), 6.93~6.98(m,lH), 6.85~6.88( m, lH), 6.05 (s, 2H), 4.19 (d, lH), 3.63 to 3.84 (m, 4H), 3.60 to 3.61 (m, lH), 3.43 to 3.46 (m, 4H), 3.31 (m, 19 95166 201228664 2H), 3.13~3.18(m,3H), 2.96~3.00(m,3H), 2.48(s,3H), 2.10~2. 13(m,2H) Example 2 (y?' Magic-2-(5-fluoro-2-oxo-1,2-dihydro-indole-3-indenyl)-5-(2-hydroxy-3-morpholin-4-yl-propyl )-3-mercapto-5, 6, 7, 8-tetrahydro-1 pico-pyrrolo[3,2-c]azepin-4-one malate

將（足幻-2-(5-氟-2-側氧基-1，2-二氫-吲哚-3-次曱基）-5-(2-羥基-3-嗎啉-4-基-丙基）-3-曱基-5, 6, 7, 8-四氫 — H11 比略並[3，2_c]0丫庚因-4-酉同 lp(469 mg，1 mmol) 攪拌下溶解於15 mL曱醇中，加入l-蘋果酸（147 mg，1. 1 mmol)。攪拌反應30分鐘，反應液減壓濃縮，加入12〇mL 乙腈，回流反應1. 5小時。冷卻至室溫，過濾，濾餅依次用冰乙腈（1 mLx3)和冰乙醇洗務（1 mLx3)，得到（及幻_2_ (5-氟-2-侧氧基-1，2-二氫-吲哚—3-次曱基）—5〜（2-經基 -3-嗎琳-4-基-丙基）-3-曱基-5, 6, 7, 8-四氫比洛並 [3, 2-c]吖庚因-4-酮蘋果酸鹽2 (535 mg，黃色固體），產率：88. 8%。 MS ra/z (ESI): 469.2[M+1] WNMRUOOMHz，fDMSO，ppm): 5 13.73(s，1H)，10.92(s， lH)，7.75~7.79(m，2H)，6.83〜6.96(m，2H)，4.9i(s，iH)， 95166 20 201228664 4. 17〜4. 20(m，1Η)’ 3·95(ιη，1H)’ 3. 72〜3.77(dd，1H)， 3.61(s，3H)，3.20〜3.37(m，4H)，2.92〜2.96(m，2H)， 2. 39〜2. 62(m, 14H), 2. 25(m，2H) 實施例3 (尤7)-2-(5-氟-2-側氧基-1，2-二氫-α弓卜朵一3一次甲基）_5_ (2-羥基-3-嗎啉-4-基-丙基）一3_曱基_5, 6, 7, 8四氫^万一 σ比p各並[3, 2-c] 〇丫庚因-4-酮乳酸鹽Will (Fantasy-2-(5-fluoro-2-indolyl-1,2-dihydro-indol-3-ylidene)-5-(2-hydroxy-3-morpholin-4-yl) -propyl)-3-indolyl-5, 6, 7, 8-tetrahydro-H11 ratio slightly [3,2_c]0丫gyne-4-indole lp (469 mg, 1 mmol) dissolved under stirring 5小时。 After cooling to room temperature, the reaction was stirred for 15 minutes, and the reaction mixture was concentrated under reduced pressure. , filtered, and the filter cake was washed with ice acetonitrile (1 mL×3) and ice ethanol (1 mL×3) to obtain (and phantom _2_(5-fluoro-2-oxooxy-1,2-dihydro-indole- 3-terpenyl)- 5~(2-carbyl-3-morphin-4-yl-propyl)-3-indolyl-5, 6, 7, 8-tetrahydropyrazine[3, 2 -c]glyoxime-4-one malate 2 (535 mg, yellow solid), yield: 88. 8%. MS s/z (ESI): 469.2[M+1] WNMRUOOMHz, fDMSO, ppm) : 5 13.73(s,1H), 10.92(s, lH), 7.75~7.79(m,2H), 6.83~6.96(m,2H),4.9i(s,iH), 95166 20 201228664 4. 17~4 . 20(m,1Η)' 3·95(ιη,1H)' 3. 72~3.77(dd,1H), 3.61(s,3H), 3.20~3.37(m,4H),2.92~2.96(m, 2H) 2. 39~2. 62(m, 14H), 2. 25(m, 2H) Example 3 (U.S. 7)-2-(5-fluoro-2-indolyl-1,2-dihydro-α弓布朵一三次 methyl)_5_(2-hydroxy-3-morpholin-4-yl-propyl)-3_mercapto_5, 6, 7, 8 tetrahydrogen ^ σ σ ratio p and [3, 2-c] azepine-4-keto lactate

將（)?，7)-2-(5-氟-2-侧氧基-1，2-二氫-吲。朵_3_次甲基）-5-(2-羥基-3-嗎琳-4-基-丙基）_3-曱基-5, 6, 7, 8-四氫-1及-吡咯並[3,2-c]吖庚因-4-酮 lp(471 mg，1 mm〇l) 攪拌下溶解於17 mL曱醇和34 mL二氯曱烷中，加入乳酸 (90 mg，1 mmol) ’攪拌反應30分鐘。減壓濃縮，加入2〇 mL乙腈，回流反應45分鐘。冷卻至室溫，過濾，滤餅依次用冰乙腈（1 mLx3)和冰乙醇洗蘇（1 mLx3)，得到 (充，^)-2-(5-|L-2-側氧基-1，2-二氫。朵-3—次曱基）-5-(2-經基-3-嗎琳-4-基-丙基）-3-曱基-5,6,7,8-四氫-1及-0比p各並[3, 2-c]a丫庚因-4-酿1乳酸鹽3(502 mg，黃色固體），產率：90%。 MS m/z (ESI): 469.2[M+1] 1HNMR(400MHz, d-dUSO, ppm): 5 13. 73(s, 1H), 10.92(s, 21 95166 201228664 6. 87(m， 1H), ，1H)，，2H)， lH)，7.75〜7.79(m，2H)，6.92〜6.94(m，lH)，684 1H)，4.74(d，1H)，3.79〜3.90(m，1H)，3.75(dd 3.59(s，3H)，3.58(m，1H)，3.32(m，2H)，3 l9(m 2.95(m, 2H), 2.46(m, 4H), 2.45(m, 8H), 2 33r 2.26(ra, 2H) ’ ’ (m 貫施例 (及，^)-2-(5-氟~2-側氧基-1，2-二氫 -5-(2-羥基-3-嗎啉一4-基-丙基）—3-甲基__5 β人甲基）各並[3’2_c]a丫庚因_4，甲磺酸鹽 1()?,7)-2-(5-fluoro-2-p-oxy-1,2-dihydro-indole. _3_methine)-5-(2-hydroxy-3-mline -4-yl-propyl)_3-mercapto-5, 6, 7, 8-tetrahydro-1 and -pyrrolo[3,2-c]azepin-4-one lp (471 mg, 1 mm 〇l) Dissolved in 17 mL of sterol and 34 mL of dichloromethane with stirring, and added lactic acid (90 mg, 1 mmol) to stir the reaction for 30 minutes. Concentrate under reduced pressure, add 2 mL of acetonitrile and reflux for 45 min. After cooling to room temperature, filtration, the filter cake was washed with ice acetonitrile (1 mL×3) and iced ethanol (1 mL×3) to give (yield:^)-2-(5-|L-2- oxo-1, 2-dihydro.-3-(indolyl)-5-(2-pyridyl-3-morphin-4-yl-propyl)-3-indolyl-5,6,7,8-tetrahydro -1 and -0 ratio p and [3, 2-c]a 丫gyne-4-broth 1 lactate 3 (502 mg, yellow solid), yield: 90%. MS m/z (ESI): 469.2 [M+1] 1H NMR (400 MHz, d-dUSO, ppm): 5 13. 73 (s, 1H), 10.92 (s, 21 95166 201228664 6. 87 (m, 1H) , , 1H), , 2H), lH), 7.75~7.79 (m, 2H), 6.92~6.94 (m, lH), 684 1H), 4.74 (d, 1H), 3.79~3.90 (m, 1H), 3.75 (dd 3.59 (s, 3H), 3.58 (m, 1H), 3.32 (m, 2H), 3 l9 (m 2.95 (m, 2H), 2.46 (m, 4H), 2.45 (m, 8H), 2 33r 2.26(ra, 2H) ' ' (m Example (and, ^)-2-(5-fluoro~2-o-oxy-1,2-dihydro-5-(2-hydroxy-3-? Phenyl- 4-yl-propyl)-3-methyl__5 β-human methyl) each [3'2_c]a agyne _4, methanesulfonate 1

將（兄^)-2-(5-氟-2-側氧基-1，2-二氫__吲哚〜3 & 基）_5_(2_經基-3-嗎琳-4_基-丙基）-3~甲武β '"曱氫-1及-吡洛並[3, 2-c]吖庚因-4-酮lp(47〇 ’8四 ’ 1 mm〇n 攪拌下溶解於8 mL甲醇和16 mL二氣甲γ中，丫，加入甲石黃酸 (96 mg，1匪〇1)，授拌反應30分鐘，減壓濃縮加入 mL乙腈’回流反應30分鐘。冷卻至室溫，過據，遽餅依次用冰乙腈（1 mLx3)和冰乙醇洗滌（1 ，得到 (尤幻-2-(5-氟-2-側氧基-1，2-二氫_吲哚_3_次甲基）_5_ (2-羥基-3-嗎啉-4-基-丙基）曱基_5, 6, 7, 8_四氫一if 吡咯並[3,2-c]吖庚因-4-酮曱磺酸鹽4(519 mg，黃色固體），產率：92%。 22 95166 201228664 MS m/z (ESI): 469.2[M+1] 1HNMR(400MHz, d-dUSO, ppm): 5 13. 77(s, 1H), 10. 94(s, 1H), 9.66(s, 1H), 7.76~7.80(m, 2H), 6. 85-6. 97(m, 2H), 5.82(s, 1H), 4.22(s, 1H), 4. 00(m, 2H), 3. 83(m, 3H), 3.77(m, 2H), 3.58-3. 61(m, 3H), 3. 21 ~3. 35(m, 4H), 3. 13(m, 2H), 2.51(s, 3H), 2. 33(s, 3H), 2. 10(m, 2H), l.l(m, 2H) 實施例5 (尤氣-2-側氧基_1，2-二氫-°引°朵-3-次曱基）-5-(2-經基-3-嗎琳-4-基-丙基）_3-曱基-5, 6, 7, 8-四氫_1及一吡咯並[3, 2-c]吖庚因-4-酮鹽酸鹽Will (brothers)-2-(5-fluoro-2-o-oxy-1,2-dihydro__吲哚~3 & base)_5_(2_yl--3-indan-4_yl -propyl)-3~甲武β '"曱H-1 and-Pylo[3,2-c]glycoin-4-one lp (47〇'8 four' 1 mm〇n under stirring Dissolved in 8 mL of methanol and 16 mL of dioxane γ, hydrazine, adding methenic acid (96 mg, 1 匪〇 1), and mixing reaction for 30 minutes, concentrated under reduced pressure and adding mL acetonitrile to reflux reaction for 30 minutes. To room temperature, the cake was washed with ice acetonitrile (1 mL×3) and iced ethanol (1 to obtain (Y-Fluor-2-(5-fluoro-2-oxooxy-1,2-dihydro-indole).哚_3_methine)_5_(2-hydroxy-3-morpholin-4-yl-propyl)indolyl_5, 6, 7, 8_tetrahydro-if-pyrrolo[3,2-c] Indomethacin-4-ketosulfonate 4 (519 mg, yellow solid), yield: 92%. 22 95166 201228664 MS m/z (ESI): 469.2 [M+1] 1HNMR (400 MHz, d-dUSO , ppm): 5 13. 77(s, 1H), 10. 94(s, 1H), 9.66(s, 1H), 7.76~7.80(m, 2H), 6. 85-6. 97(m, 2H ), 5.82(s, 1H), 4.22(s, 1H), 4. 00(m, 2H), 3. 83(m, 3H), 3.77(m, 2H), 3.58-3. 61(m, 3H ), 3. 21 ~ 3. 35 (m, 4H), 3. 13 (m, 2H), 2.51 (s, 3H), 2. 33 (s, 3H), 2. 10 (m, 2H), ll(m, 2H) Example 5 (European-2-sideoxy-1,2-dihydro-° ̄ ̄-3--3-indolyl)-5-(2- Benzyl-3-morphin-4-yl-propyl)_3-mercapto-5, 6, 7, 8-tetrahydro-1 and pyrrolo[3,2-c]azepin-4-one Acid salt

將U，^)-2-(5-氟-2-側氧基-1，2-二氫-吲哚-3-次甲基）-5-(2-羥基-3-嗎啉-4-基-丙基）-3-曱基-5, 6, 7, 8-四氫-1皮_〇比洛並[3, 2-c]吖庚因-4-酮 lp (486 mg，1 mmol) 攪拌下溶解於5 mL曱醇和8 mL二氣曱烷中，加入2 mL 5M 氯化氫的1,4-二氧六環溶液，搜拌反應1小時。減壓濃縮，加入50 mL乙腈，回流反應1小時。冷卻至室溫，過濾，得到U, 7)-2-(5-氟-2-側氧基-1，2-二氫-吲哚-3-次曱基）-5-(2-經基-3-嗎淋_4-基-丙基）-3-曱基-5, 6, 7, 8-四氫-1)7-吡咯並[3, 2-d吖庚因—4-酮鹽酸鹽5 (459 mg，黃 23 95166 201228664 色固體），產率：91%。 MS m/z (ESI): 469.2[M+1] ^NMRGOOMHz，d-DMSO，ppm): (5 13.73(s，1H)，10.91(s, lH)，7.76〜7.78(m，lH)，7.75(s，lH)，6.91〜6.94(m，lH)， 6.84〜6.87(m，lH)，4.72〜4.73(d，lH)，3.90(m，lH)， 3.75〜3.79(dd，lH)，3.57〜3.59(t，4H)，3.38〜3.35(t， 2H)，3.14〜3.19(dd，lH)，2.92〜2.95(t，2H)，2.46(s，3H)， 2.42〜2.51(m，4H)，2.29〜2.31(t，2H)，2.08(m,2H) 測試例：溶解度測試按照常規溶解度測定方法，測試式（I)化合物及其鹽在生理鹽水中的溶解度，結果如表1所示：表1 : 鹽形式生理鹽水式（I)化合物 0.00186 實施例1 0. 1214 實施例2 0.0227 實施例3 0.00124 實施例4 0.0131 實施例5 0.0294 結論：式（I)化合物的馬來酸鹽與其游離鹼及其它鹽相比溶解度明顯改善。藥代動力學測試測試例1 本發明的化合物藥代動力學測試 1、試驗目的 24 95166 201228664 以大鼠為受試動物，應用LC/MS/MS法測定了大鼠分別經灌胃給予式（I)化合物及其不同鹽及尾靜脈注射給予式（I)化合物馬來酸鹽後不同時刻血漿中的藥物濃度。研究本發明的化合物在大鼠體内的藥代動力學行為，評價其藥動學特徵，並考察其口服絕對生物利用度。 2、試驗方案 2. 1、試驗藥品式（I)化合物，實施例1至5化合物 2. 2、試驗動物健康成年SD大鼠2 8隻’雖雄各半，平均分成7組每組4隻，購自上海西普爾-必凱實驗動物有限公司，動物生產許可證號：SCXK(滬）2008-0016。 2. 3、儀器設備 TSQ Quantum Ultra AM三重四極桿質譜儀，美國Therm〇 Finnigan 公司；U,^)-2-(5-fluoro-2-p-oxy-1,2-dihydro-indol-3-methyl)-5-(2-hydroxy-3-morpholine-4- --propyl)-3-mercapto-5, 6, 7, 8-tetrahydro-1 pi 〇洛洛并 [3, 2-c] 吖heptin-4-one lp (486 mg, 1 mmol Dissolved in 5 mL of decyl alcohol and 8 mL of dioxane under stirring, and added 2 mL of 5 M hydrogen chloride in 1,4-dioxane solution, and the reaction was stirred for 1 hour. It was concentrated under reduced pressure, and 50 mL of acetonitrile was added, and the mixture was refluxed for 1 hour. Cool to room temperature and filter to give U, 7)-2-(5-fluoro-2-oxooxy-1,2-dihydro-indole-3-indenyl)-5-(2-yl) -3-Olin- 4-yl-propyl)-3-indolyl-5, 6, 7, 8-tetrahydro-1)7-pyrrolo[3,2-d吖g-in 4-one salt Acid salt 5 (459 mg, yellow 23 95166 201228664 color solid), yield: 91%. MS m/z (ESI): 469.2 [M+1], NMR, s, s, s, s, s, s, s, s, s, s, s, s, s, s, s, s, (s, lH), 6.91~6.94 (m, lH), 6.84~6.87 (m, lH), 4.72~4.73 (d, lH), 3.90 (m, lH), 3.75~3.79 (dd, lH), 3.57 ~3.59(t,4H), 3.38~3.35(t, 2H), 3.14~3.19(dd,lH), 2.92~2.95(t,2H), 2.46(s,3H), 2.42~2.51(m,4H) 2.29 to 2.31 (t, 2H), 2.08 (m, 2H) Test Example: Solubility Test The solubility of the compound of the formula (I) and its salt in physiological saline was tested according to the conventional solubility measurement method, and the results are shown in Table 1: Table 1: Salt form physiological saline Compound (I) 0.00186 Example 1 0. 1214 Example 2 0.0227 Example 3 0.00124 Example 4 0.0131 Example 5 0.0294 Conclusion: Maleate of the compound of formula (I) and its free base Solubility improved significantly compared to other salts. Pharmacokinetic test test example 1 Pharmacokinetic test of the compound of the present invention 1. Test objective 24 95166 201228664 Rats were used as test animals, and were determined by LC/MS/MS method. Rats were given formula (I) by gavage The concentration of the drug in plasma at different times after administration of the compound and its different salts and tail vein injection of the maleate salt of the compound of formula (I). The pharmacokinetic behavior of the compound of the present invention in rats was evaluated and its pharmacokinetics was evaluated. Characteristics, and examine its oral absolute bioavailability. 2. Test protocol 2. 1. Test drug formula (I) compound, examples 1 to 5 compound 2. 2. Test animals healthy adult SD rats 2 8 ' The average half is divided into 7 groups of 4 each, purchased from Shanghai Xipuer-Beikai Experimental Animal Co., Ltd., animal production license number: SCXK (Shanghai) 2008-0016. 2. 3, instrument equipment TSQ Quantum Ultra AM triple Quadrupole mass spectrometer, Therm〇Finnigan Company, USA;

Agilent 1200高效液相色譜系統，美國Agilent公司。 2. 4、藥物配製靜脈注射組：稱取適量藥物，加入生理鹽水溶解並稀釋至終體積，使樣品濃度為1.0 mg/mL。灌胃給藥組：稱取適量藥物’加入0 5% CMC_Na超聲製成1. 0 mg/mL混懸液，臨用時配製並注意避光。 2· 5、給藥健康成年SD大鼠28隻，雌雄各半，平均分成7組，每組4隻。禁食過夜後分別灌胃給藥或尾靜脈注射給予實 95166 25 201228664 施例1化合物，給藥劑量均為10 mg/kg (以域基部分計），給藥體積10 mL/kg。 2. 6、樣品採集靜脈注射給藥組於給藥前及給藥後2 min, 15 min，30 min，1.0 h，2. Oh, 4. Oh, 6.0 h，8. 0h，12. Oh, 24. 0 h，36· 0 h由眼眶採血0. 2 mL，置於肝素化試管中，3500 rpm 離心10分鐘分離血漿，於-20°C保存。灌胃給藥組於給藥前及給藥後0.5，1.0，2.0，3.0， 4.0，6.0，8.0，12.0，24. 0，36. Oh 採血，樣品處理方法同靜脈注射給藥組。給藥後2h進食。 2. 7、分析方法取給藥後各時刻的大鼠血蒙25 /Z L，加入内標溶液20 //L’曱醇125" L，渦旋混合2分鐘，離心10分鐘（16000 r/min)，取上清液i〇#l進行LC-MS/MS分析。 2.8、標準曲線製備取大鼠空白血漿25 y L，分別加入標準系列溶液，使血藥濃度為 1.00，2.00，5.00，25.0，100，500，2000， 5000 ng/mL，加入内標溶液2〇从l，甲醇1〇〇以L ,按“血漿樣品預處理’’項下進行操作。以血藥濃度為橫坐標，樣品與内標色譜峰面積比為縱坐標，以加權最小二乘法 (w=l/x2)進行線性回歸，獲得典型標準曲線方程。 2.9、藥代動力學參數計算對夂試化合物的藥代動力學行為進行房室模型擬合，並計算主要藥代動力學參數’其中Cmax、七ax採用實測值。 95166 26 201228664 根據灌胃及尾靜脈注射給藥後AUCo-t計算口服絕對生物利用度。 3、藥代動力學參數結果本發明的化合物的藥代動力學參數如表2所示。結論：式（I)化合物的馬來酸鹽與游離鹼及其它鹽相比，藥代動力學性質和生物利用度明顯改善，具有明顯的藥代動力學優勢。表2 : 化合物 F (%) CL (//g/mL) AUdx (/zg-h/mL) t 1/2 (h) TL (h) MRT (h) CL/F (mL/min/kg) 式⑴ 化合物 27.9 0. 64±0.24 4.49 土 2.79 3.31±0.57 2. 00±1.41 5.24+2. 23 1.84±1.20 靜脈 16.12±3.97 5. 97±2.50 — 1.40+0.35 0. 65±0.161 實施例1 48.2 0. 95±0.37 7.77±2.99 3.01±0.60 5.75±2. 06 6.66±2. 05 1.44+0.55 靜脈 16.12±3.97 5. 97+2.50 — 1.40±0. 35 0. 65+0.161 實施例2 30.4 0.85±0.39 4.90+2.79 3.13+0.97 1.75±1.66 4.29+1.87 1.79±0.67 靜脈 16.12+3.97 5· 97±2· 50 — 1.40+0.35 0.65+0.161 實施例3 36.6 0.81±0.45 5.90+3.35 3.06+1.21 2.50±1.00 6.28+1. 56 1. 64+0. 70 靜脈 16.12±3.97 5.97+2.50 — 1.40+0. 35 0. 65±0.161 實施例4 45.3 0. 77+0.39 7.30±5,87 3.41±0.85 3.75±1.71 6. 68+1.58 1.49+0. 69 靜脈 16.12+3.97 5. 97+2.50 — 1.40±0.35 0. 65±0.161 實施例5 25.2 0.50±0.10 4.07±1.63 3. 60+0.52 4. 25+2. 06 6.80+1.58 2. 03+0. 51 靜脈 16.12±3.97 5. 97±2.50 — 1.40+0.35 0.65±0.161 【圖式簡單說明】無【主要元件符號說明】無 27 95166Agilent 1200 High Performance Liquid Chromatography System, Agilent, USA. 2. Preparation of the drug Intravenous group: Weigh the appropriate amount of the drug, dissolve it in physiological saline and dilute it to the final volume to make the sample concentration 1.0 mg/mL. The gavage administration group: Weighed the appropriate amount of the drug 'Addition of 0 5% CMC_Na ultrasound to make a 1.0 mg / mL suspension, prepared and taken care of in the dark. 2. 5, administration 28 healthy adult SD rats, male and female, divided into 7 groups, 4 in each group. After fasting overnight, the administration of the compound of Example 1 was administered by intragastric administration or tail vein injection at a dose of 10 mg/kg (based on the domain basis) at a dose of 10 mL/kg. 2. 6. Sample collection intravenous administration group before administration and 2 minutes after administration, 15 min, 30 min, 1.0 h, 2. Oh, 4. Oh, 6.0 h, 8. 0h, 12. Oh, 24. 0 h, 36 · 0 h Blood was collected from the eyelids by 0.2 mL, placed in heparinized tubes, centrifuged at 3500 rpm for 10 minutes, and stored at -20 °C. The rats in the gavage administration group were administered with 0.5, 1.0, 2.0, 3.0, 4.0, 6.0, 8.0, 12.0, 24.0, Oh. before the administration and after the administration, and the sample treatment method was the same as the intravenous administration group. Eat 2 hours after administration. 2. Analytical method Take 25/ZL of rat blood at each time after administration, add internal standard solution 20 //L' sterol 125 " L, vortex for 2 minutes, centrifuge for 10 minutes (16000 r/min) ), taking the supernatant i〇#l for LC-MS/MS analysis. 2.8. Preparation of standard curve Take 25 y L of rat blank plasma and add standard series solution to make blood concentration 1.00, 2.00, 5.00, 25.0, 100, 500, 2000, 5000 ng/mL, add internal standard solution 2〇 From l, methanol 1 〇〇 to L, according to the "plasma sample pretreatment"' operation. With the blood concentration as the abscissa, the ratio of the sample to the internal standard chromatographic peak area is the ordinate, with weighted least squares method (w =l/x2) Perform a linear regression to obtain a typical standard curve equation. 2.9. Pharmacokinetic parameter calculations Fitting the pharmacokinetic behavior of the test compound with the atrioventricular model and calculating the main pharmacokinetic parameters' Cmax and seven ax were measured. 95166 26 201228664 Oral absolute bioavailability was calculated according to AUCo-t after intragastric and tail vein administration. 3. Pharmacokinetic parameter results The pharmacokinetic parameters of the compounds of the present invention are as follows: Table 2 shows that the maleate salt of the compound of formula (I) has significantly improved pharmacokinetic properties and bioavailability compared with the free base and other salts, and has obvious pharmacokinetic advantages. Table 2: Compound F (%) CL (//g/mL) AUdx (/zg-h/mL) t 1/2 (h) TL (h) MRT (h) CL/F (mL/min/kg) Formula (1) Compound 27.9 0. 64±0.24 4.49 Soil 2.79 3.31±0.57 2. 00±1.41 5.24+2. 23 1.84±1.20 Vein 16.12±3.97 5. 97±2.50 — 1.40+0.35 0. 65±0.161 Example 1 48.2 0. 95 ±0.37 7.77±2.99 3.01±0.60 5.75±2. 06 6.66±2. 05 1.44+0.55 Vein 16.12±3.97 5. 97+2.50 — 1.40±0. 35 0. 65+0.161 Example 2 30.4 0.85±0.39 4.90+ 2.79 3.13+0.97 1.75±1.66 4.29+1.87 1.79±0.67 Vein 16.12+3.97 5·97±2· 50 — 1.40+0.35 0.65+0.161 Example 3 36.6 0.81±0.45 5.90+3.35 3.06+1.21 2.50±1.00 6.28+1 56 1. 64+0. 70 Vein 16.12±3.97 5.97+2.50 — 1.40+0. 35 0. 65±0.161 Example 4 45.3 0. 77+0.39 7.30±5,87 3.41±0.85 3.75±1.71 6. 68 +1.58 1.49+0. 69 Vein 16.12+3.97 5. 97+2.50 — 1.40±0.35 0. 65±0.161 Example 5 25.2 0.50±0.10 4.07±1.63 3. 60+0.52 4. 25+2. 06 6.80+1.58 2. 03+0. 51 Vein 16.12±3.97 5. 97±2.50 — 1.40+0.35 0.65±0.161 [Simple description of the diagram] No [Main component symbol description] 2795166

Claims

201228664 七、申請專利範圍： 1. 一種式（I)所示的（无，^)-2-(5-氟-2-側氧基2〜一知 π弓卜朵-3-次曱基）-5-(2-經基-3-嗎啉·_4_基一丙基）〜3 ^ 基-5, 6, 7, 8m㈣並[3, 2_C]D丫庚因 + 藥用的鹽：201228664 VII. Patent application scope: 1. A formula (I) (n, ^)-2-(5-fluoro-2-sided oxy 2~1 π π 卜 -3- 曱 )) -5-(2-carbyl-3-morpholine·_4_yl-propyl)~3^yl-5, 6, 7, 8m(tetra) and [3, 2_C]D丫gyne + pharmaceutically acceptable salts:

==範圍第2項所述的鹽’其中該無機鹽選二 I鹽、鹽酸鹽、硫酸鹽、硝酸鹽或氫溴酸鹽。 2.二請專利範圍p項所述的鹽，其中該鹽為 4·:申請專利範圍第2項所述的鹽，其中該;機鹽為鹽酸 5.如申請專利範圍第丨項所述的鹽，其晻 I ㈣5娜 %酸鹽、馬來酸鹽、酒石酸鹽、琥⑽鹽、^趟Μ $醋酸鹽、富馬酸鹽、檸檬酸鹽、枸櫞酸鹽、笨磺：二本甲酸鹽、萘磺酸鹽、乳酸鹽或蘋果酸鹽。 I 7· 利範圍第5項所述的鹽，其中‘機鹽選自頻如酸果酸鹽、孔酸鹽、曱磺酸鹽或馬來酸鹽。二請專利範圍第5項所述㈣，其中料機鹽為馬來種製備申請專利範圍第】至8項中任〜項所述鹽的方 95166 1 201228664 法’該方法包括將（亿^)-2-(5-氟-2-側氧基2_- 1 °引°木-3-次曱基）-5-(2-沒基-3-嗎琳-4-基-内基）_3一甲基-5, 6, 7, 8-四氫-1及-«比略並[3, 2-c]吖庚因_4_酮與相應的酸成鹽的步驟。 10·如申請專利範圍第9項所述的方法，其中該酸為選自鱗酸、鹽酸、硫酸、硝酸、氫溴酸、曱磺酸、馬來酸、酒石酸、琥珀酸、醋酸、三氟醋酸、富馬酸、擰檬酸、枸梅酸、苯績酸、苯甲酸、萘石黃酸、导L酸或頻^酸的無機酸或有機酸。 11. 一種藥物組合物，其含有治療有效劑量的申請專利範圍第1至8項中任一項所述的鹽以及可藥用的載體。已 12·—種申請專利範圍第丨至8項中任一項所述的鹽在製備治療與蛋白質激酶有關的疾病的藥物中的用途。 13· —種申請專利範圍第11項所述的藥物組合物在製備治療與蛋白質激與有關的疾病的藥物中的用途。 14. 如申請專利範圍第12項或第13項所述的用途，其令該與蛋白質激酶有關的疾病選自與VEGFR-2，EGFR HER-2, PDGFR，C-Kit，c-Met 或 FGFR 相關的疾病。 15. 如申請專利顧第14項所述的⑽，其中該疾病為癌症。山 16.如申請專利範圍第15項所述的用途，其中該癌症選自肺癌、乳腺癌、表皮鱗癌或胃癌。 K-㈣請專利範㈣項中任一項所述的鹽在製備蛋白質激酶抑制劑的藥物中的用途，其中該蛋白激酶選 95166 2 201228664 自 VEGFR-2， EGFR， HER-2， PDGFR， c-Kit， c-Met 或 FGFR。 18. —種申請專利範圍第11項所述的藥物組合物在製備蛋白質激酶抑制劑的藥物中的用途，其中該蛋白激酶選自 VEGFR-2, EGFR，HER-2, PDGFR，c-Kit，c-Met 或 FGFR。 201228664 四、指定代表圖：本案無圖式 (一) 本案指定代表圖為：第（）圖。 (二) 本代表圖之元件符號簡單說明：五、本案若有化學式時，請揭示最能顯示發明特徵的化學式：== The salt described in the above item 2 wherein the inorganic salt is selected from a salt, a hydrochloride, a sulfate, a nitrate or a hydrobromide. 2. The salt according to the scope of claim p, wherein the salt is 4: the salt described in claim 2, wherein the organic salt is hydrochloric acid 5. As described in the scope of the patent application Salt, its dark I (tetra) 5 na% acid salt, maleate salt, tartrate salt, amber (10) salt, ^ 趟Μ $ acetate, fumarate, citrate, citrate, stupid: two Acid salt, naphthalene sulfonate, lactate or malate. I. The salt of item 5, wherein the 'organic salt is selected from the group consisting of a salt such as a citrate, a porate, an oxime sulfonate or a maleate. 2. Please refer to the fourth paragraph (4) of the patent scope, in which the salt of the machine is the salt of the Malay species, the preparation of the patent scope 】 to the salt of any of the eight items of the item 95166 1 201228664 method 'The method includes (100 million) -2-(5-fluoro-2-indolyl 2_- 1 ° 引木-3--3-indolyl)-5-(2-diyl-3-morphin-4-yl-endyl)_3 A step of forming a salt of methyl-5,6,7,8-tetrahydro-1 and -« succinyl[3,2-c]oxime _4-ketone with the corresponding acid. 10. The method of claim 9, wherein the acid is selected from the group consisting of scaly acid, hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, sulfonic acid, maleic acid, tartaric acid, succinic acid, acetic acid, and trifluorocarbon. An inorganic or organic acid of acetic acid, fumaric acid, citric acid, phthalic acid, benzoic acid, benzoic acid, naphthoic acid, L-acid or acid. A pharmaceutical composition comprising a therapeutically effective amount of the salt of any one of claims 1 to 8 and a pharmaceutically acceptable carrier. The use of a salt according to any one of claims 1 to 8 for the preparation of a medicament for treating a protein kinase-related disease. 13. Use of the pharmaceutical composition of claim 11 in the preparation of a medicament for treating a disease associated with protein stimulation. 14. The use of the protein kinase-related disease selected from the group consisting of VEGFR-2, EGFR HER-2, PDGFR, C-Kit, c-Met or FGFR, as claimed in claim 12 or claim 13 Related diseases. 15. (10) as claimed in claim 14, wherein the disease is cancer. The use according to claim 15, wherein the cancer is selected from the group consisting of lung cancer, breast cancer, epidermal squamous cell carcinoma or gastric cancer. K-(d) The use of a salt according to any one of the patents, wherein the protein kinase is selected from the group consisting of VEGFR-2, EGFR, HER-2, PDGFR, c -Kit, c-Met or FGFR. Use of the pharmaceutical composition according to claim 11 for the preparation of a protein kinase inhibitor, wherein the protein kinase is selected from the group consisting of VEGFR-2, EGFR, HER-2, PDGFR, c-Kit, c-Met or FGFR. 201228664 IV. Designated representative map: There is no schema in this case (1) The representative representative map of this case is: (). (2) A brief description of the symbol of the representative figure: 5. If there is a chemical formula in this case, please disclose the chemical formula that best shows the characteristics of the invention:

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