TW201808948A - Imidazo[4,5-c]quinolin-2-one compounds and their use in treating cancer - Google Patents

Imidazo[4,5-c]quinolin-2-one compounds and their use in treating cancer Download PDF

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TW201808948A
TW201808948A TW106108065A TW106108065A TW201808948A TW 201808948 A TW201808948 A TW 201808948A TW 106108065 A TW106108065 A TW 106108065A TW 106108065 A TW106108065 A TW 106108065A TW 201808948 A TW201808948 A TW 201808948A
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compound
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pharmaceutically acceptable
methyl
imidazo
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伯納德 克里斯多夫 巴蘭
科特 高登 派克
湯瑪士 安東尼 杭特
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阿斯特捷利康公司
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Abstract

The specification generally relates to compounds of Formula (I): and pharmaceutically acceptable salts thereof, where R1 is tetrahydropyran-3-yl or 3,3-dimethyltetrahydropyran-4-yl; R2 is methyl or hydro; R3 is hydro or fluoro; R4 is hydro or fluoro; and R5 is methyl or hydro; the use of compounds of Formula (I) or pharmaceutically acceptable salts thereof to treat or prevent ATM mediated disease, including cancer; pharmaceutical compositions comprising substituted imidazo[4,5-c]quinolin-2-one compounds or pharmaceutically acceptable salts thereof; kits comprising compounds of Formula (I) or pharmaceutically acceptable salts thereof; methods of manufacture of compounds of Formula (I) or pharmaceutically acceptable salts thereof; and intermediates useful in such manufacture.

Description

咪唑并[4,5-c]喹啉-2-酮化合物以及它們在治療癌症中之用途    Imidazo [4,5-c] quinolin-2-one compounds and their use in the treatment of cancer   

本說明書涉及經取代的咪唑并[4,5-c]喹啉-2-酮化合物及其藥學上可接受的鹽。該等化合物和鹽選擇性地調節共濟失調毛細血管擴張症突變的(“ATM”)激酶，並且因此本說明書還涉及經取代的咪唑并[4,5-c]喹啉-2-酮化合物及其鹽治療或預防ATM介導的疾病(包括癌症)之用途。本說明書還涉及包含經取代的咪唑并[4,5-c]喹啉-2-酮化合物及其鹽的醫藥組成物；包含此類化合物和鹽之套組(kit)；生產此類化合物和鹽之方法；以及在此類生產中有用的中間體。 This specification relates to substituted imidazo [4,5-c] quinolin-2-one compounds and pharmaceutically acceptable salts thereof. These compounds and salts selectively modulate ataxia telangiectasia mutated ("ATM") kinases, and therefore this specification also relates to substituted imidazo [4,5-c] quinolin-2-one compounds And its salts for the treatment or prevention of ATM-mediated diseases (including cancer). This specification also relates to pharmaceutical compositions containing substituted imidazo [4,5-c] quinolin-2-one compounds and their salts; kits containing such compounds and salts; producing such compounds and Salt methods; and intermediates useful in such production.

ATM激酶係絲胺酸蘇胺酸激酶，最初鑒定為在共濟失調毛細血管擴張症中的突變基因的產物。共濟失調毛細血管擴張症位於人染色體11q22-23上並且編碼約350kDa的一個大蛋白質，其由磷脂醯肌醇(“PI”)3-激酶樣絲胺酸/蘇胺酸激酶結構域的存在來表徵，該結構域由調節ATM激酶活性和功能的FRAP-ATM-TRRAP結構域和FATC結構域側翼。ATM激酶已被鑒定為藉由雙股斷裂引起的DNA損傷應答的主要參與者。它主要在 S/G2/M細胞週期過渡中並在坍塌複製叉處起作用以引發細胞週期檢查點、染色質修飾、HR修復以及促存活信號級聯放大，以便在DNA損傷後保持細胞完整性(拉文(Lavin)，M.F.；分子細胞生物學綜述(Rev.Mol.Cell Biol.) 2008，759-769)。 The ATM kinase is serine threonine kinase, originally identified as the product of a mutant gene in ataxia telangiectasia. Ataxic telangiectasia is located on human chromosome 11q22-23 and encodes a large protein of approximately 350 kDa, which is caused by the presence of the phospholipid inositol ("PI") 3-kinase-like serine / threonine kinase domain To characterize, this domain is flanked by FRAP-ATM-TRRAP and FATC domains that regulate ATM kinase activity and function. ATM kinase has been identified as a major player in the response to DNA damage caused by double-strand breaks. It mainly functions in the S / G2 / M cell cycle transition and at the collapse replication fork to initiate cell cycle checkpoints, chromatin modification, HR repair, and pro-survival signal cascade amplification to maintain cell integrity after DNA damage (Lavin, MF; Review of Molecular Cell Biology (Rev. Mol. Cell Biol. 2008 , 759-769).

ATM激酶傳訊大致可分為兩類：典型途徑，該途徑與來自雙股斷裂的Mre11-Rad50-NBS1複合物在一起發信號並啟動DNA損傷檢查點；和活化的若干非典型模式，該等模式藉由其他形式的細胞應激被啟動(克雷莫納(Cremona)等人，癌基因(Oncogene)2013，3351-3360)。 ATM kinase signaling can be roughly divided into two categories: the typical pathway, which signals the Mre11-Rad50-NBS1 complex from the double-strand break and initiates DNA damage checkpoints; and several atypical modes of activation, these modes It is activated by other forms of cellular stress (Cremona et al., Oncogene 2013 , 3351-3360).

ATM激酶迅速地、強勁地被啟動以響應於雙股斷裂，且據說能夠在過量的800種底物中磷酸化(松崗(Matsuoka)等人，科學(Science)2007，1160-1166)，協調多個應激反應途徑(庫爾茲(Kurz)和利斯米爾(Lees Miller)，DNA修復(DNA Repair)2004，889-900)。ATM激酶以無活性同型二聚體形式主要存在於細胞的細胞核中，但在感測到DNA雙股斷裂(典型途徑)時在Ser1981上自磷酸化，導致具有全激酶活性的單體的解離(貝克漢尼斯特(Bakkenist)等人，自然(Nature)2003，499-506)。這係一個關鍵的啟動事件，並且因此針對腫瘤途徑依賴性，ATMphospho-Ser1981係直接藥效學的和患者的選擇生物標誌物兩者。 ATM kinase is rapidly and strongly activated in response to double-strand breaks and is said to be phosphorylated in excess of 800 substrates (Matsuoka et al., Science 2007 , 1160-1166), coordination Multiple stress response pathways (Kurz and Lees Miller, DNA Repair 2004 , 889-900). ATM kinase is mainly present in the nucleus of cells in the form of inactive homodimer, but it is self-phosphorylated on Ser1981 when a double strand break of DNA is sensed (typical pathway), resulting in the dissociation of monomers with full kinase activity Bakkenist et al., Nature 2003 , 499-506). This is a key initiating event, and therefore directed at tumor pathway dependence, ATMphospho-Ser1981 is both a direct pharmacodynamic and patient-selected biomarker.

ATM激酶響應於由常見抗癌治療如電離輻射和拓撲異構酶-II抑制劑(多柔比星(doxorubicin)、依託泊苷(etoposide))所造成的直接的雙股斷裂，而且藉由複製過程中的單股斷裂至雙 股斷裂轉換還響應於拓撲異構酶-I抑制劑(例如伊立替康(irinotecan)和托泊替康(topotecan))。ATM激酶抑制可以增強任何該等試劑的活性，並且結果係ATM激酶抑制劑預期在癌症的治療中係有用的。 ATM kinase responds to direct double-strand breaks caused by common anti-cancer treatments such as ionizing radiation and topoisomerase-II inhibitors (doxorubicin, etoposide), and by replication The single-strand break to double-strand break transition in the process is also responsive to topoisomerase-I inhibitors (such as irinotecan and topotecan). ATM kinase inhibition can enhance the activity of any of these agents, and as a result ATM kinase inhibitors are expected to be useful in the treatment of cancer.

CN 102372711 A報導了某些咪唑并[4,5-c]喹啉-2-酮化合物，該等化合物被稱為PI 3-激酶α和哺乳動物雷帕黴素(rapamycin)靶蛋白(“mTOR”)激酶的雙重抑制劑： CN 102372711 A reports certain imidazo [4,5-c] quinolin-2-one compounds, which are called PI 3-kinase alpha and mammalian rapamycin target protein ("mTOR ") Dual inhibitors of kinases:

在CN 102372711 A中報導的某些化合物Certain compounds reported in CN 102372711 A

CN 102399218 A報導了某些咪唑并[4,5-c]喹啉-2-酮化合物，該等化合物稱為PI 3-激酶α抑制劑： CN 102399218 A reports certain imidazo [4,5-c] quinolin-2-one compounds, which are called PI 3-kinase alpha inhibitors:

在CN 102399218 A中報導的某些化合物Certain compounds reported in CN 102399218 A

雖然CN 102372711 A和CN 102399218 A的該等化合物被報導具有對抗PI 3-激酶α並且在某些情況下對抗mTOR激酶的活性，但對研發更有效對抗不同激酶(如ATM激酶)的新化合物仍存在需求。對以高度選擇性方式(即，藉由比其他生物靶標更有效地調節ATM)作用於某些激酶(像ATM激酶)的新化合物進一步存在需求。 Although these compounds of CN 102372711 A and CN 102399218 A have been reported to have activity against PI 3-kinase α and in some cases against mTOR kinase, they are still effective in the development of new compounds that are more effective against different kinases (such as ATM kinase) There is a demand. There is a further need for new compounds that act on certain kinases (like ATM kinases) in a highly selective manner (ie, by modulating ATM more efficiently than other biological targets).

如在本說明書中別處(例如在實驗部分中描述的基於細胞的測定中)證明的，本說明書的該等化合物通常具有非常強的ATM激酶抑制活性，但對其他酪胺酸激酶，如PI 3-激酶α、 mTOR激酶、共濟失調毛細血管擴張症和Rad3-相關蛋白(“ATR”)激酶、以及DNA依賴性蛋白激酶(“DNAPK”)具有小得多的活性。因此，本說明書的該等化合物不僅抑制ATM激酶，還可以被認為係ATM激酶的高度選擇性抑制劑。 As demonstrated elsewhere in this specification (e.g. in cell-based assays described in the experimental section), the compounds of this specification generally have very strong ATM kinase inhibitory activity, but against other tyrosine kinases, such as PI 3 -Kinase alpha, mTOR kinase, ataxia telangiectasia and Rad3-related protein ("ATR") kinase, and DNA-dependent protein kinase ("DNAPK") have much less activity. Therefore, the compounds of this specification not only inhibit ATM kinase, but can also be considered as a highly selective inhibitor of ATM kinase.

作為其高度選擇性性質的結果，本說明書的該等化合物預期在ATM激酶牽連於其中的疾病的治療中(例如，在癌症的治療中)特別有用，但其中希望的是最小化由於其他酪胺酸激酶，如類PI 3-激酶α、mTOR激酶、ATR激酶和/或DNAPK的抑制可能產生的脫靶作用或毒性。 As a result of their highly selective nature, the compounds of this specification are expected to be particularly useful in the treatment of diseases in which ATM kinases are implicated (eg, in the treatment of cancer), but where it is desirable to minimize Inhibition of acid kinases, such as PI-like 3-kinase α, mTOR kinase, ATR kinase, and / or DNAPK, may produce off-target effects or toxicity.

簡言之，本說明書部分地描述了具有化學式(I)之化合物： In short, this specification partially describes the compound of formula (I):

或其藥學上可接受的鹽，其中：R 1 係四氫哌喃-3-基或3,3-二甲基四氫哌喃-4-基；R 2 係甲基或氫；R 3 係氫或氟；R 4 係氫或氟；並且 R 5 係甲基或氫。 Or a pharmaceutically acceptable salt thereof, wherein: R 1 is tetrahydropiperan-3-yl or 3,3-dimethyltetrahydropiperan-4-yl; R 2 is methyl or hydrogen; R 3 is Hydrogen or fluorine; R 4 is hydrogen or fluorine; and R 5 is methyl or hydrogen.

本說明書還部分地描述了包括具有化學式(I)之化合物或其藥學上可接受的鹽，以及至少一種藥學上可接受的賦形劑的醫藥組成物。 This specification also partially describes a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

本說明書還部分地描述了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在療法中使用。 This specification also partially describes a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in therapy.

本說明書還部分地描述了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用。 This specification also partially describes a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer.

本說明書還部分地描述了具有化學式(I)之化合物或其藥學上可接受的鹽在生產用於治療癌症的藥物中之用途。 This specification also partially describes the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating cancer.

本說明書還部分地描述了用於在需要這種治療的溫血動物中治療癌症之方法，該方法包括向所述溫血動物給予治療有效量的具有化學式(I)之化合物或其藥學上可接受的鹽。 This specification also partially describes a method for treating cancer in a warm-blooded animal in need of such treatment, the method comprising administering to the warm-blooded animal a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable Accept the salt.

[圖1]：(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮之形式A之X-射線粉末繞射圖。 [ Figure 1 ]: ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H -X-ray powder diffraction pattern of Form A of imidazo [4,5-c] quinolin-2 (3H) -one.

[圖2]：(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮之形式A之DSC熱譜圖。 [ Figure 2 ]: ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H -DSC thermogram of form A of imidazo [4,5-c] quinolin-2 (3H) -one.

[圖3]：(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基 -1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式A之X-射線粉末繞射圖。 [ Figure 3 ]: ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3- Radical) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form A X-ray powder diffraction pattern.

[圖4]：(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式A之DSC熱譜圖。 [ Figure 4 ]: ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3- Group) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form A DSC thermogram.

[圖5]：(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式B之X-射線粉末繞射圖。 [ Figure 5 ]: ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3- Radical) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one Form B X-ray powder diffraction pattern.

[圖6]：(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式B之DSC熱譜圖。 [ Figure 6 ]: ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3- Radical) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form B DSC thermogram.

許多實施方式在本說明書中詳細描述，並且對於本領域有技術的讀者而言將是明顯的。該等實施方式不被解釋為限制性的。 Many embodiments are described in detail in this specification, and will be apparent to those skilled in the art. These embodiments are not to be interpreted as limiting.

在第一個實施方式中，提供了具有化學式(I)之化合物： 或其藥學上可接受的鹽，其中：R 1 係四氫哌喃-3-基或3,3-二甲基四氫哌喃-4-基；R 2 係甲基或氫；R 3 係氫或氟；R 4 係氫或氟；並且R 5 係甲基或氫。 In a first embodiment, a compound of formula (I) is provided: Or a pharmaceutically acceptable salt thereof, wherein: R 1 is tetrahydropiperan-3-yl or 3,3-dimethyltetrahydropiperan-4-yl; R 2 is methyl or hydrogen; R 3 is Hydrogen or fluorine; R 4 is hydrogen or fluorine; and R 5 is methyl or hydrogen.

“氫”基團相當於氫原子。其上附接氫基團的原子可被認定為係未經取代的。 The "hydrogen" group corresponds to a hydrogen atom. The atom to which the hydrogen group is attached may be regarded as unsubstituted.

術語“藥學上可接受的”通常指定一個物件(例如鹽、劑型或賦形劑)係適合在患者中使用的。藥學上可接受的鹽的實例清單可以發現於：藥用鹽手冊：性質、選擇和使用(the Handbook of Pharmaceutical Salts：Properties，Selection and Use)，P.H.斯塔爾(Stahl)和C.G.韋穆特(Wermuth)編輯，魏因海姆(Weinheim)/蘇黎世(Zürich)：威利(Wiley)-VCH出版社/VHCA，2002。具有化學式(I)之化合物的適合的藥學上可接受的鹽例如是酸加成鹽。在技術人員已知的條件下，具有化學式(I)之化合物的酸加成鹽可以藉由使該化合物與適合的無機酸或有機酸接觸來形成。酸加成鹽例如可以使用選自鹽酸、氫溴酸、硫酸和磷酸的無機酸來形成。酸加成鹽還可以使用選自以下各項的有機酸來形成：三氟乙酸、檸檬酸、馬來酸、草酸、乙酸、甲酸、苯甲酸、富馬酸、琥珀酸、酒石酸、乳酸、丙酮酸、甲磺酸、苯磺酸以及對甲苯磺酸。 The term "pharmaceutically acceptable" generally designates an article (eg, salt, dosage form, or excipient) that is suitable for use in a patient. A list of examples of pharmaceutically acceptable salts can be found in: The Handbook of Pharmaceutical Salts: Properties, Selection and Use (The Handbook of Pharmaceutical Salts: Properties, Selection and Use ), PH Stahl and CG Vemut ( Edited by Wermuth, Weinheim / Zürich: Wiley-VCH Press / VHCA, 2002. Suitable pharmaceutically acceptable salts of compounds of formula (I) are, for example, acid addition salts. Under conditions known to the skilled person, an acid addition salt of a compound of formula (I) can be formed by contacting the compound with a suitable inorganic or organic acid. The acid addition salt can be formed using, for example, an inorganic acid selected from hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid. Acid addition salts can also be formed using organic acids selected from the group consisting of trifluoroacetic acid, citric acid, maleic acid, oxalic acid, acetic acid, formic acid, benzoic acid, fumaric acid, succinic acid, tartaric acid, lactic acid, acetone Acid, methanesulfonic acid, benzenesulfonic acid and p-toluenesulfonic acid.

因此，在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，其中該藥學上可接受的鹽係鹽酸鹽、氫溴酸鹽、硫酸鹽、磷酸鹽、三氟乙酸鹽、檸檬酸鹽、馬來酸鹽、草酸鹽、乙酸鹽、甲酸鹽、苯甲酸鹽、富馬酸鹽、琥珀酸鹽、酒石酸鹽、乳酸鹽、丙酮酸鹽、甲磺酸鹽、苯磺酸鹽或對甲苯磺酸鹽。在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，其中該藥學上可接受的鹽係甲磺酸鹽。在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，其中該藥學上可接受的鹽係單-甲磺酸鹽，即具有化學式(I)之化合物對甲磺酸的化學計量係1：1。 Therefore, in one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided, wherein the pharmaceutically acceptable salt is hydrochloride, hydrobromide, sulfate, phosphate, Trifluoroacetate, citrate, maleate, oxalate, acetate, formate, benzoate, fumarate, succinate, tartrate, lactate, pyruvate, formate Sulfonate, besylate or p-toluenesulfonate. In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein the pharmaceutically acceptable salt is a mesylate salt. In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided, wherein the pharmaceutically acceptable salt is a mono-methanesulfonate salt, that is , the compound of formula (I) is The stoichiometry of sulfonic acid is 1: 1.

另一個實施方式提供了本文所定義的任何實施方式(例如如申請專利範圍1所述的實施方式)，其條件係選自實例1、2、3、4、5、6、7、8、9、10、11和12的一個或多個具體的實例(例如一個、兩個或三個具體的實例)單獨地被放棄。 Another embodiment provides any of the embodiments defined herein (for example, the embodiments described in patent scope 1), the conditions of which are selected from Examples 1, 2, 3, 4, 5, 6, 7, 8, 9. One, or more specific instances (e.g., one, two, or three specific instances) of, 10, 11, and 12 are individually discarded.

化學式(I)中的可變基團的一些值如下。該等值可以與任何定義、申請專利範圍(例如申請專利範圍第1項)、或本文所定義的實施方式組合使用以提供另外的實施方式。 Some values of the variable group in chemical formula (I) are as follows. These values can be used in combination with any definition, patent application scope (for example, patent application scope item 1), or the embodiments defined herein to provide additional embodiments.

a)R 1 係四氫哌喃-3-基。 a) R 1 is tetrahydropiperan-3-yl.

b)R 1 係(S)-四氫哌喃-3-基。 b) R 1 series ( S ) -tetrahydropiperan-3-yl.

c)R 1 係(R)-四氫哌喃-3-基。 c) R 1 series ( R ) -tetrahydropiperan-3-yl.

d)R 1 係3,3-二甲基四氫哌喃-4-基。 d) R 1 is 3,3-dimethyltetrahydropiperan-4-yl.

e)R 1 係(S)-3,3-二甲基四氫哌喃-4-基。 e) R 1 series ( S ) -3,3-dimethyltetrahydropiperan-4-yl.

f)R 1 係(R)-3,3-二甲基四氫哌喃-4-基。 f) R 1 series ( R ) -3,3-dimethyltetrahydropiperan-4-yl.

g)R 2 係甲基。 g) R 2 is methyl.

h)R 2 係氫。 h) R 2 is hydrogen.

i)R 3 係氫。 i) R 3 is hydrogen.

j)R 3 係氟。 j) R 3 is fluorine.

k)R 4 係氫。 k) R 4 is hydrogen.

l)R 4 係氟。 l) R 4 is fluorine.

m)R 5 係甲基。 m) R 5 is methyl.

n)R 5 係氫。 n) R 5 is hydrogen.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，其中：R 1 係四氫哌喃-3-基；R 2 係甲基或氫；R 3 係氫或氟；R 4 係氫或氟；並且R 5 係甲基。 In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided, wherein: R 1 is tetrahydropiperan-3-yl; R 2 is methyl or hydrogen; R 3 is hydrogen Or fluorine; R 4 is hydrogen or fluorine; and R 5 is methyl.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，其中：R 1 係3,3-二甲基四氫哌喃-4-基； R 2 係甲基或氫；R 3 係氫或氟；R 4 係氫或氟；並且R 5 係甲基。 In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided, wherein: R 1 is 3,3-dimethyltetrahydropiperan-4-yl; R 2 is methyl Or hydrogen; R 3 is hydrogen or fluorine; R 4 is hydrogen or fluorine; and R 5 is methyl.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，其中：R 1 係3,3-二甲基四氫哌喃-4-基；R 2 係甲基；R 3 係氫；R 4 係氫；並且R 5 係甲基。 In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided, wherein: R 1 is 3,3-dimethyltetrahydropiperan-4-yl; R 2 is methyl ; R 3 is hydrogen; R 4 is hydrogen; and R 5 is methyl.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，其中該化合物選自：(R)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮；(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮；(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮；(R)-7-氟-8-(2-氟-6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫 -2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮；(S)-7-氟-8-(2-氟-6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮；(R)-8-(2-氟-6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮；(S)-8-(2-氟-6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮；(R)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮；外消旋-1-(3,3-二甲基四氫-2H-哌喃-4-基)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮；(R)-1-(3,3-二甲基四氫-2H-哌喃-4-基)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮；(S)-1-(3,3-二甲基四氫-2H-哌喃-4-基)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮；以及(S)-8-(6-(甲氧基甲基)吡啶-3-基)-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein the compound is selected from: ( R ) -8- (6- (methoxymethyl) pyridine-3- Yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5-c] quinolin-2 (3H) -one; ( S ) -8 -(6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5-c] Quinoline-2 (3H) -one; ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H- Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one; ( R ) -7-fluoro-8- (2-fluoro-6- (Methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5-c] quinoline-2 (3H) -one; ( S ) -7-fluoro-8- (2-fluoro-6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H- Piperan-3-yl) -1H-imidazo [4,5-c] quinolin-2 (3H) -one; ( R ) -8- (2-fluoro-6- (methoxymethyl) pyridine -3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5-c] quinolin-2 (3H) -one; ( S ) -8- (2-fluoro-6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5-c] quinoline-2 (3H) -one; ( R ) -7-fluoro-8 -(6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5-c] Quinoline-2 (3H) -one; racemic-1- (3,3-dimethyltetrahydro-2H-piperan-4-yl) -8- (6- (methoxymethyl) pyridine -3-yl) -3-methyl-1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one; ( R ) -1- (3,3-dimethyl Tetrahydro-2H-piperan-4-yl) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1,3-dihydro-2H-imidazo [4 , 5-c] quinolin-2-one; ( S ) -1- (3,3-dimethyltetrahydro-2H-piperan-4-yl) -8- (6- (methoxymethyl ) Pyridin-3-yl) -3-methyl-1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one; and ( S ) -8- (6- (methyl Oxymethyl) pyridin-3-yl) -1- (tetrahydro-2H-piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinoline-2 -ketone.

在一個實施方式中，提供了具有化學式(I)之任何化合物或其藥學上可接受的鹽，它們可以根據實例部分中的實驗細節來製備。 In one embodiment, there is provided any compound of formula (I) or a pharmaceutically acceptable salt thereof, which can be prepared according to experimental details in the example section.

在一個實施方式中，提供了(S)-8-(6-(甲氧基甲基)吡 啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮或其藥學上可接受的鹽。 In one embodiment, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl ) -1H-imidazo [4,5-c] quinolin-2 (3H) -one or a pharmaceutically acceptable salt thereof.

在一個實施方式中，提供了(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮。 In one embodiment, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl ) -1H-imidazo [4,5-c] quinolin-2 (3H) -one.

在一個實施方式中，提供了(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮的藥學上可接受的鹽。 In one embodiment, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl ) -1H-imidazo [4,5-c] quinolin-2 (3H) -one pharmaceutically acceptable salt.

在一個實施方式中，提供了(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮或其藥學上可接受的鹽。 In one embodiment, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan is provided -3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one or a pharmaceutically acceptable salt thereof.

在一個實施方式中，提供了(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮。 In one embodiment, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan is provided -3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one.

在一個實施方式中，提供了(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮的藥學上可接受的鹽。 In one embodiment, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan is provided -3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one is a pharmaceutically acceptable salt.

在一個實施方式中，提供了(S)-8-(6-(甲氧基甲基)吡啶-3-基)-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮或其藥學上可接受的鹽。 In one embodiment, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -1- (tetrahydro-2H-piperan-3-yl) -1,3 is provided -Dihydro-2H-imidazo [4,5-c] quinolin-2-one or a pharmaceutically acceptable salt thereof.

在一個實施方式中，提供了(S)-8-(6-(甲氧基甲基)吡啶-3-基)-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉 -2-酮。 In one embodiment, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -1- (tetrahydro-2H-piperan-3-yl) -1,3 is provided -Dihydro-2H-imidazo [4,5-c] quinolin-2-one.

在一個實施方式中，提供了(S)-8-(6-(甲氧基甲基)吡啶-3-基)-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮的藥學上可接受的鹽。 In one embodiment, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -1- (tetrahydro-2H-piperan-3-yl) -1,3 is provided -A pharmaceutically acceptable salt of dihydro-2H-imidazo [4,5-c] quinolin-2-one.

本說明書中描述的化合物和鹽能以溶劑化形式和非溶劑化形式存在。例如，溶劑化形式可以是水合形式，如半水合物、一水合物、二水合物、三水合物或其可替代的數量。本發明涵蓋具有化學式(I)之化合物的所有該等溶劑化和非溶劑化形式，具體地是該等形式具有ATM激酶抑制活性的程度，如例如使用本文所描述的測試測量的。 The compounds and salts described in this specification can exist in solvated and unsolvated forms. For example, the solvated form may be a hydrated form, such as hemihydrate, monohydrate, dihydrate, trihydrate, or an alternative amount thereof. The present invention encompasses all such solvated and unsolvated forms of compounds of formula (I) , specifically the extent to which these forms have ATM kinase inhibitory activity, as measured, for example, using the tests described herein.

本說明書所描述的該等化合物和鹽的原子可以作為它們的同位素存在。本發明涵蓋了具有化學式(I)之所有化合物，其中原子被其同位素中的一個或多個替換(例如具有化學式(I)之化合物，其中一個或多個碳原子係11C或13C碳同位素，或其中一個或多個氫原子係2H或3H同位素)。 The atoms of the compounds and salts described in this specification may exist as their isotopes. The present invention covers all compounds of formula (I) in which atoms are replaced by one or more of their isotopes (eg compounds of formula (I) in which one or more carbon atoms are 11 C or 13 C carbon isotopes , Or one or more hydrogen atom system 2 H or 3 H isotopes).

本說明書中所描述的化合物和鹽可以按互變異構物的混合物存在。“互變異構物”係結構異構物，其存在於由氫原子的遷移產生的平衡中。本發明包括具有化學式(I)之化合物的所有互變異構物，具體地是該等互變異構物具有ATM激酶抑制活性的程度。 The compounds and salts described in this specification may exist as a mixture of tautomers. "Tautomers" are structural isomers, which exist in equilibrium resulting from the migration of hydrogen atoms. The present invention includes all tautomers of the compound of formula (I) , specifically the degree to which the tautomers have ATM kinase inhibitory activity.

本說明書中所描述的化合物和鹽借助不對稱碳原子而以光學活性形式或外消旋形式存在。本發明包括具有化學式(I) 之化合物的任何光學活性形式或外消旋形式，所述形式具有ATM激酶抑制活性，如例如使用本文所描述的測試測量的。光學活性形式的合成可以藉由本領域中熟知的有機化學標準技術進行，例如藉由使用光學活性物質合成或藉由拆分外消旋形式。 The compounds and salts described in this specification exist in an optically active form or a racemic form by means of asymmetric carbon atoms. The present invention includes any optically active form or racemic form of the compound of formula (I) having ATM kinase inhibitory activity, as measured, for example, using the tests described herein. The synthesis of optically active forms can be carried out by standard techniques of organic chemistry well known in the art, for example by synthesis using optically active substances or by resolution of racemic forms.

因此，在一個實施方式中，提供了具有化學式(I)之化合物、或其藥學上可接受的鹽，其係鏡像異構物過量(%ee)95%、98%或99%的單一光學異構物。在一個實施方式中，單一光學異構物以鏡像異構物過量(%ee)99%存在。 Therefore, in one embodiment, there is provided a compound of formula (I) , or a pharmaceutically acceptable salt thereof, which is an enantiomeric excess (% ee) 95%, 98% or 99% of single optical isomers. In one embodiment, the single optical isomer is in excess of the mirror image isomer (% ee) 99% exists.

在一個實施方式中，提供了具有化學式(I)之化合物、或其藥學上可接受的鹽，其係鏡像異構物過量(%ee)95%、98%或99%的(S)-光學異構物。在一個實施方式中，(S)-光學異構物以鏡像異構物過量(%ee)99%存在。 In one embodiment, there is provided a compound of formula (I) , or a pharmaceutically acceptable salt thereof, which is an enantiomeric excess (% ee) 95%, 98% or 99% of ( S ) -optical isomers. In one embodiment, the ( S ) -optical isomer is in excess of the mirror image isomer (% ee) 99% exists.

在一個實施方式中，提供了具有化學式(I)之化合物、或其藥學上可接受的鹽，其係鏡像異構物過量(%ee)95%、98%或99%的(R)-光學異構物。在一個實施方式中，(R)-光學異構物以鏡像異構物過量(%ee)99%存在。 In one embodiment, there is provided a compound of formula (I) , or a pharmaceutically acceptable salt thereof, which is an enantiomeric excess (% ee) 95%, 98% or 99% of ( R ) -optical isomers. In one embodiment, the ( R ) -optical isomer is in excess of the mirror image isomer (% ee) 99% exists.

本說明書中所描述的化合物和鹽可以是晶體，並且可以展示一種或多種晶體形式。本發明涵蓋具有化學式(I)之化合物的任何晶體或非晶形形式，或該等形式的混合物，都具有ATM激酶抑制活性。 The compounds and salts described in this specification may be crystalline, and may exhibit one or more crystal forms. The present invention covers any crystalline or amorphous form of the compound of formula (I) , or a mixture of these forms, which has ATM kinase inhibitory activity.

通常已知可以使用常規技術表徵結晶物質，如X射線粉末繞射(XRPD)、差示掃描熱量測定(DSC)、熱解重量分析 (TGA)、漫反射紅外傅裡葉變換(DRIFT)光譜法、近紅外(NIR)光譜法、溶液和/或固態核磁共振光譜法。該等結晶物質的水含量可以藉由卡爾費歇爾分析(Karl Fischer analysis)測定。 It is generally known that conventional techniques can be used to characterize crystalline materials, such as X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), diffuse reflection infrared Fourier transform (DRIFT) spectroscopy , Near infrared (NIR) spectroscopy, solution and / or solid-state nuclear magnetic resonance spectroscopy. The water content of these crystalline substances can be determined by Karl Fischer analysis.

本文所描述的該等晶體形式提供了基本上與附圖中示出的XRPD圖相同的XRPD圖，並且具有如本文中包括的表中所示出的不同的2-θ值。熟習該項技術者將理解的是，可以獲得取決於測量條件(如所用設備或機器)而具有一個或多個測量誤差的XRPD圖或繞射圖。類似地，通常已知的是，XRPD圖中的強度可取決於測量條件或樣品製備作為較佳的是取向的結果波動。XRPD領域的普通技術人員將進一步認識到，峰的相對強度還可以受例如大小在30μm以上的晶粒和非單一縱橫比影響。技術人員理解的是，反射位置可以受樣品在繞射計中所處的確切高度和繞射計的零點校正影響。樣品的表面平坦度也可能具有細微影響。 The crystal forms described herein provide an XRPD pattern that is substantially the same as the XRPD pattern shown in the drawings, and have different 2-theta values as shown in the tables included herein. Those skilled in the art will understand that an XRPD pattern or a diffraction pattern with one or more measurement errors depending on the measurement conditions (such as the equipment or machine used) can be obtained. Similarly, it is generally known that the intensity in the XRPD pattern can fluctuate depending on the measurement conditions or sample preparation as the result of the orientation is preferably. One of ordinary skill in the art of XRPD will further realize that the relative intensity of peaks can also be affected by, for example, grains with a size above 30 μm and a non-single aspect ratio. The skilled person understands that the reflection position can be influenced by the exact height of the sample in the diffractometer and the zero-point correction of the diffractometer. The surface flatness of the sample may also have a slight effect.

作為該等考慮的結果，所呈現的繞射圖數據不應視為絕對值(詹金斯R(Jenkins,R)和辛德爾R.L.(Snyder,R.L.)《X射線粉末繞射測定法的介紹》(‘Introduction to X-Ray Powder Diffractometry’)，約翰．威利父子公司(John Wiley & Sons)1996；邦C.W.(Bunn,C.W.)(1948)，《化學晶體學》(Chemical Crystallography)，倫敦克拉倫登出版社(Clarendon Press,London)；克盧格H.P.(Klug,H.P.)和亞歷山大L.E.(Alexander,L.E.)(1974)，《X射線繞射程序》(‘X-Ray Diffraction Procedures’))。應當對應地理解的是，所述固體形式不局限於提供與附圖中所示的XRPD圖相同的XRPD圖的晶體，並且任何提供 與附圖中所示的那些基本相同的XRPD圖的晶體落入本發明的範圍內。XRPD領域的技術人員能夠判斷XRPD圖的實質一致性。通常，在XRPD中的繞射角的測量誤差為約±0.2°2-θ，並且當考慮附圖中的X射線粉末繞射圖時，並且當讀取包含於其中的表中包含的數據時，這樣的測量誤差的程度應該考慮在內。 As a result of these considerations, the diffraction data presented should not be regarded as absolute values (Jenkins R (Jenkins, R) and Sinder RL (Snyder, RL) "Introduction to X-ray powder diffraction measurement method" (' Introduction to X-Ray Powder Diffractometry '), John Wiley & Sons 1996; CW (Bunn, CW) (1948), Chemical Crystallography, Clarendon Publishing, London (Clarendon Press, London); Klug HP (Klug, HP) and Alexander LE (Alexander, LE) (1974), "X-Ray Diffraction Procedures" ('X-Ray Diffraction Procedures'). It should be correspondingly understood that the solid form is not limited to crystals that provide the same XRPD pattern as the XRPD patterns shown in the drawings, and any crystal that provides XRPD patterns that are substantially the same as those shown in the figures It is within the scope of the present invention. A person skilled in the field of XRPD can judge the substantial consistency of the XRPD chart. In general, the measurement error of the diffraction angle in XRPD is about ± 0.2 ° 2-θ, and when considering the X-ray powder diffraction diagram in the drawings, and when reading the data contained in the table contained therein The degree of such measurement error should be taken into account.

實例2的化合物展現出晶體特性，並且一晶體形式已經被表徵。 The compound of Example 2 exhibits crystalline properties, and a crystalline form has been characterized.

因此，在一個實施方式中，提供了(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮之形式A。 Therefore, in one embodiment, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3 -Yl) -1H-imidazo [4,5-c] quinolin-2 (3H) -one, Form A.

在一個實施方式中，提供了晶體形式，(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮之形式A，它的X射線粉末繞射圖在約2-θ=7.0°處具有至少一個特異峰。 In one embodiment, a crystalline form is provided, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan- 3-yl) -1H-imidazo [4,5-c] quinoline-2 (3H) -one form A, its X-ray powder diffraction pattern has at least one specificity at about 2-θ = 7.0 ° peak.

在一個實施方式中，提供了晶體形式，(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮之形式A，它的X射線粉末繞射圖在約2-θ=9.2°處具有至少一個特異峰。 In one embodiment, a crystalline form is provided, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan- 3-yl) -1H-imidazo [4,5-c] quinoline-2 (3H) -one form A, its X-ray powder diffraction pattern has at least one specificity at about 2-θ = 9.2 ° peak.

在一個實施方式中，提供了晶體形式，(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮之形式A，它的X射線粉末繞射圖在約2-θ=7.0°和9.2°處具有至少兩個特異峰。 In one embodiment, a crystalline form is provided, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan- 3-yl) -1H-imidazo [4,5-c] quinoline-2 (3H) -one form A, its X-ray powder diffraction pattern has at about 2-θ = 7.0 ° and 9.2 ° At least two specific peaks.

在一個實施方式中，提供了晶體形式，(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮之形式A，它的X射線粉末繞射圖在約2-θ=7.0°、9.2°、10.4°、13.9°、18.9°、22.3°、23.1°、23.6°、24.7°、和25.4°處具有特異峰。 In one embodiment, a crystalline form is provided, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan- 3-yl) -1H-imidazo [4,5-c] quinoline-2 (3H) -one, Form A, its X-ray powder diffraction pattern is about 2-θ = 7.0 °, 9.2 °, 10.4 There are specific peaks at °, 13.9 °, 18.9 °, 22.3 °, 23.1 °, 23.6 °, 24.7 °, and 25.4 °.

在一個實施方式中，提供了晶體形式，(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮之形式A，它的X射線粉末繞射圖與圖1中所示的X射線粉末繞射圖係基本上相同的。 In one embodiment, a crystalline form is provided, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan- 3-yl) -1H-imidazo [4,5-c] quinoline-2 (3H) -one form A, its X-ray powder diffraction pattern and the X-ray powder diffraction pattern shown in FIG. 1 Department is basically the same.

在一個實施方式中，提供了晶體形式，(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮之形式A，它的X射線粉末繞射圖在2-θ=7.0°±0.2° 2-θ處具有至少一個特異峰。 In one embodiment, a crystalline form is provided, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan- 3-yl) -1H-imidazo [4,5-c] quinoline-2 (3H) -one, Form A, its X-ray powder diffraction pattern is at 2-θ = 7.0 ° ± 0.2 ° 2-θ There is at least one specific peak.

在一個實施方式中，提供了晶體形式，(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮之形式A，它的X射線粉末繞射圖在2-θ=9.2°±0.2° 2-θ處具有至少一個特異峰。 In one embodiment, a crystalline form is provided, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan- 3-yl) -1H-imidazo [4,5-c] quinoline-2 (3H) -one, Form A, its X-ray powder diffraction pattern is at 2-θ = 9.2 ° ± 0.2 ° 2-θ There is at least one specific peak.

在一個實施方式中，提供了晶體形式，(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮之形式A，它的X射線粉末繞射圖在2-θ=7.0°和9.2°±0.2° 2-θ處具有至少兩個特異峰。 In one embodiment, a crystalline form is provided, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan- 3-yl) -1H-imidazo [4,5-c] quinolin-2 (3H) -one, Form A, its X-ray powder diffraction pattern is at 2-θ = 7.0 ° and 9.2 ° ± 0.2 ° At 2-θ there are at least two specific peaks.

在一個實施方式中，提供了晶體形式，(S)-8-(6-(甲 氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮之形式A，它的X射線粉末繞射圖在2-θ=7.0°、9.2°、10.4°、13.9°、18.9°、22.3°、23.1°、23.6°、24.7°、和25.4°±0.2° 2-θ處具有特異峰。 In one embodiment, a crystalline form is provided, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan- 3-yl) -1H-imidazo [4,5-c] quinolin-2 (3H) -one, Form A, its X-ray powder diffraction pattern is at 2-θ = 7.0 °, 9.2 °, 10.4 ° , 13.9 °, 18.9 °, 22.3 °, 23.1 °, 23.6 °, 24.7 °, and 25.4 ° ± 0.2 ° have specific peaks at 2-θ.

(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮之形式A的DSC分析顯示以184.4℃開始的熔融吸熱和在186.0℃處的峰(圖2)。 ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4 The DSC analysis of Form A of, 5-c] quinoline-2 (3H) -one showed an endotherm of melting starting at 184.4 ° C and a peak at 186.0 ° C (Figure 2).

熟習該項技術者理解，在具體的化合物的DSC熱譜圖中觀察到的值或值的範圍將顯示不同純度的批次之間的變化。因此，儘管對於一種化合物而言該範圍可以是小的，但是對於其他化合物而言該範圍可以是相當大的。通常，DSC熱事件中的繞射角的測量誤差係約正負5℃，並且當考慮包含於其中的DSC數據時，這樣的測量誤差的程度應該考慮在內。 Those skilled in the art understand that the values or ranges of values observed in the DSC thermogram of a specific compound will show variations between batches of different purity. Therefore, although the range may be small for one compound, it may be quite large for other compounds. Generally, the measurement error of the diffraction angle in a DSC thermal event is about plus or minus 5 ° C, and when considering the DSC data contained therein, the degree of such measurement error should be taken into account.

因此，在一個實施方式中，提供了晶體形式，(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮之形式A，其具有以約184.4℃開始的熔融和在約186.0℃處的峰的DSC吸熱。 Therefore, in one embodiment, a crystalline form is provided, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piper (An-3-yl) -1H-imidazo [4,5-c] quinolin-2 (3H) -one, Form A, which has a melting starting at about 184.4 ° C and a DSC of the peak at about 186.0 ° C Endothermic.

因此，在一個實施方式中，提供了晶體形式，(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮之形式A，其具有以184.4℃±5℃開始的熔融和在186.0℃±5℃處的峰的DSC吸熱。 Therefore, in one embodiment, a crystalline form is provided, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piper Form-3-A) -1H-imidazo [4,5-c] quinoline-2 (3H) -one, which has a melting starting at 184.4 ° C ± 5 ° C and at 186.0 ° C ± 5 ° C The DSC of the peak is endothermic.

在一個實施方式中，提供了晶體形式，(S)-8-(6-(甲 氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮之形式A，其具有以184.4℃開始的熔融和在186.0℃處的峰的DSC吸熱。 In one embodiment, a crystalline form is provided, ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan- 3-yl) -1H-imidazo [4,5-c] quinolin-2 (3H) -one, Form A, which has melting starting at 184.4 ° C and DSC endotherm of the peak at 186.0 ° C.

在一個實施方式中，提供了晶體形式，(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮之形式A，其具有基本上如圖2中所示的DSC熱譜圖。 In one embodiment, a crystalline form is provided, (S) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan- 3-yl) -1H-imidazo [4,5-c] quinolin-2 (3H) -one, Form A, which has a DSC thermogram substantially as shown in FIG. 2.

實例3的化合物展現出晶體特性，並且兩種晶體形式已經被表徵。 The compound of Example 3 exhibits crystal characteristics, and two crystal forms have been characterized.

因此，在一個實施方式中，提供了(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式A。 Therefore, in one embodiment, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H- Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form A.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式A，它的X射線粉末繞射圖在約2-θ=4.6°處具有至少一個特異峰。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form A, its X-ray powder diffraction pattern is about 2-θ = 4.6 ° has at least one specific peak.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式A，它的X射線粉末繞射圖在約2-θ=16.0°處具有至少一個特異峰。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form A, its X-ray powder diffraction pattern is about 2-θ There is at least one specific peak at = 16.0 °.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3- 二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式A，它的X射線粉末繞射圖在約2-θ=4.6°和16.0°處具有至少兩個特異峰。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form A, its X-ray powder diffraction pattern is about 2-θ = 4.6 ° and 16.0 ° have at least two specific peaks.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式A，它的X射線粉末繞射圖在約2-θ=4.6°、8.5°、9.3°、13.8°、16.0°、18.1°、18.7°、19.9°、23.1°、24.5°處具有特異峰。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form A, its X-ray powder diffraction pattern is about 2-θ = 4.6 °, 8.5 °, 9.3 °, 13.8 °, 16.0 °, 18.1 °, 18.7 °, 19.9 °, 23.1 °, 24.5 ° have specific peaks.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式A，它的X射線粉末繞射圖與圖3中所示的X射線粉末繞射圖係基本上相同的。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form A, its X-ray powder diffraction pattern is shown in Figure 3 The X-ray powder diffraction pattern shown is basically the same.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式A，它的X射線粉末繞射圖在2-θ=4.6°±0.2° 2-θ處具有至少一個特異峰。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form A, its X-ray powder diffraction pattern is at 2-θ = 4.6 ° ± 0.2 ° There is at least one specific peak at 2-θ.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式A，它的X射線粉末繞射圖在2-θ=16.0°±0.2° 2-θ處具有至少一個特異峰。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form A, its X-ray powder diffraction pattern is at 2-θ = 16.0 ° ± 0.2 ° There is at least one specific peak at 2-θ.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式A，它的X射線粉末繞射圖 在2-θ=4.6°和16.0°±0.2° 2-θ處具有至少兩個特異峰。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form A, its X-ray powder diffraction pattern is at 2-θ = There are at least two specific peaks at 4.6 ° and 16.0 ° ± 0.2 ° 2-θ.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式A，它的X射線粉末繞射圖在2-θ=4.6°、8.5°、9.3°、13.8°、16.0°、18.1°、18.7°、19.9°、23.1°、24.5°±0.2° 2-θ處具有特異峰。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form A, its X-ray powder diffraction pattern is at 2-θ = There are specific peaks at 4.6 °, 8.5 °, 9.3 °, 13.8 °, 16.0 °, 18.1 °, 18.7 °, 19.9 °, 23.1 °, 24.5 ° ± 0.2 ° 2-θ.

(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式A的DSC分析顯示以174.5℃開始的熔融吸熱和在177.3℃處的峰(圖4)。 ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1, DSC analysis of the form A of 3-dihydro-2H-imidazo [4,5-c] quinolin-2-one showed a melting endotherm starting at 174.5 ° C and a peak at 177.3 ° C (Figure 4).

因此，在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式A，其具有以約174.5℃開始的熔融和在約177.3℃處的峰的DSC吸熱。 Therefore, in one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro -2H-piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one in Form A, which has a melting starting at about 174.5 ° C and The DSC of the peak at about 177.3 ° C is endothermic.

因此，在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式A，其具有以174.5℃±5℃開始的熔融和在177.3℃±5℃處的峰的DSC吸熱。 Therefore, in one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro -2H-piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form A, which has a melting starting at 174.5 ° C ± 5 ° C And the DSC endotherm of the peak at 177.3 ° C ± 5 ° C.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式A，其具有以174.5℃開始的熔融和在177.3℃處的峰的DSC吸熱。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form A, which has a melting starting at 174.5 ° C and at 177.3 ° C The DSC of the peak is endothermic.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式A，其具有基本上如圖4中所示的DSC熱譜圖。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form A, which has a DSC heat substantially as shown in FIG. 4 Spectra.

在一個實施方式中，提供了(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式B。 In one embodiment, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan is provided -3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form B.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式B，它的X射線粉末繞射圖在約2-θ=8.9°處具有至少一個特異峰。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form B, its X-ray powder diffraction pattern is about 2-θ = 8.9 ° has at least one specific peak.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式B，它的X射線粉末繞射圖在約2-θ=22.0°處具有至少一個特異峰。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form B, its X-ray powder diffraction pattern is about 2-θ = 22.0 ° has at least one specific peak.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式B，它的X射線粉末繞射圖在約2-θ=8.9°和22.0°處具有至少兩個特異峰。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form B, its X-ray powder diffraction pattern is about 2-θ There are at least two specific peaks at = 8.9 ° and 22.0 °.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式B，它的X射線粉末繞射圖 在約2-θ=8.9°、10.1°、11.0°、11.9°、13.9°、16.2°、16.5°、20.3°、22.0°和25.9°處具有特異峰。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form B, its X-ray powder diffraction pattern is about 2-θ There are specific peaks at = 8.9 °, 10.1 °, 11.0 °, 11.9 °, 13.9 °, 16.2 °, 16.5 °, 20.3 °, 22.0 ° and 25.9 °.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式B，它的X射線粉末繞射圖與圖5中所示的X射線粉末繞射圖係基本上相同的。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form B, its X-ray powder diffraction pattern is shown in Figure 5 The X-ray powder diffraction pattern shown is basically the same.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式B，它的X射線粉末繞射圖在2-θ=8.9°±0.2° 2-θ處具有至少一個特異峰。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form B, its X-ray powder diffraction pattern is at 2-θ = There is at least one specific peak at 8.9 ° ± 0.2 ° 2-θ.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式B，它的X射線粉末繞射圖在2-θ=22.0°±0.2° 2-θ處具有至少一個特異峰。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form B, its X-ray powder diffraction pattern is at 2-θ = 22.0 ° ± 0.2 ° There is at least one specific peak at 2-θ.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式B，它的X射線粉末繞射圖在2-θ=8.9°和22.0°±0.2° 2-θ處具有至少兩個特異峰。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form B, its X-ray powder diffraction pattern is at 2-θ = There are at least two specific peaks at 8.9 ° and 22.0 ° ± 0.2 ° 2-θ.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式B，它的X射線粉末繞射圖在2-θ=8.9°、10.1°、11.0°、11.9°、13.9°、16.2°、16.5°、20.3°、 22.0°和25.9°±0.2° 2-θ處具有特異峰。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form B, its X-ray powder diffraction pattern is at 2-θ = There are specific peaks at 8.9 °, 10.1 °, 11.0 °, 11.9 °, 13.9 °, 16.2 °, 16.5 °, 20.3 °, 22.0 ° and 25.9 ° ± 0.2 ° 2-θ.

(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式B的DSC分析顯示以174.5℃開始的熔融吸熱和在175.5℃處的峰(圖6)。 ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1, DSC analysis of the form B of 3-dihydro-2H-imidazo [4,5-c] quinolin-2-one showed a melting endotherm starting at 174.5 ° C and a peak at 175.5 ° C (Figure 6).

因此，在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式B，其具有以約174.5℃開始的熔融和在約175.5℃處的峰的DSC吸熱。 Therefore, in one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro -2H-piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form B, which has a melting starting at about 174.5 ° C and The DSC endotherm of the peak at about 175.5 ° C.

因此，在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式B，其具有以174.5℃±5℃開始的熔融和在175.5℃±5℃處的峰的DSC吸熱。 Therefore, in one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro -2H-piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form B, which has a melting starting at 174.5 ° C ± 5 ° C And the DSC endotherm of the peak at 175.5 ° C ± 5 ° C.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式B，其具有以174.5℃開始的熔融和在175.5℃處的峰的DSC吸熱。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form B, which has a melting starting at 174.5 ° C and at 175.5 ° C The DSC of the peak is endothermic.

在一個實施方式中，提供了晶體形式，(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮之形式B，其具有基本上如圖6中所示的DSC熱譜圖。 In one embodiment, a crystal form is provided, ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H -Piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one, Form B, which has a DSC heat substantially as shown in FIG. 6 Spectra.

當指出實施方式涉及晶體形式時，結晶度可大於約 60%。在一些實施方式中，結晶度大於約80%。在一些實施方式中，結晶度大於約90%。在一些實施方式中，結晶度大於約95%。在一些實施方式中，結晶度大於約98%。 When it is pointed out that the embodiment relates to a crystal form, the crystallinity may be greater than about 60%. In some embodiments, the crystallinity is greater than about 80%. In some embodiments, the crystallinity is greater than about 90%. In some embodiments, the crystallinity is greater than about 95%. In some embodiments, the crystallinity is greater than about 98%.

當指出實施方式涉及晶體形式時，結晶度可大於60%。在一些實施方式中，結晶度大於80%。在一些實施方式中，結晶度大於90%。在一些實施方式中，結晶度大於95%。在一些實施方式中，結晶度大於98%。 When it is pointed out that the embodiment relates to a crystal form, the crystallinity may be greater than 60%. In some embodiments, the crystallinity is greater than 80%. In some embodiments, the crystallinity is greater than 90%. In some embodiments, the crystallinity is greater than 95%. In some embodiments, the crystallinity is greater than 98%.

具有化學式(I)之化合物例如可以藉由具有化學式(II)之化合物： The compound of formula (I) can be, for example, a compound of formula (II) :

或其鹽(其中R 1 、R 2 以及R 3 係如本文任何實施方式中所定義的，並且X係離去基團(例如鹵素原子，或可替代地是氟原子))與具有化學式(III)之化合物： Or a salt thereof (wherein R 1, R 2 and R 3 line such as any of the embodiments defined herein, and X-based leaving group (e.g. a halogen atom, or alternatively a fluorine atom)) having the formula (III ) Compound:

或其鹽(其中R 4 和R 5 係如本文任何實施方式中所定義的，並且Y係硼酸、硼酸酯或三氟硼酸鉀基團(例如硼酸、硼酸頻哪醇(pinacol)酯、或三氟硼酸鉀))進行反應來製備。該反應可以在 熟習該項技術者熟知的標準條件下進行，例如在鈀來源(例如四合三苯基膦鈀或乙酸鈀(II))、視情況膦配位基(例如Xantphos或S-phos)、以及適合的鹼(例如碳酸銫或三乙胺)存在下。 Or a salt thereof (wherein R 4 and R 5 are as defined in any of the embodiments herein, and Y is a boric acid, borate or potassium trifluoroborate group (eg boric acid, pinacol borate, or Potassium trifluoroborate)) was prepared by reaction. The reaction can be carried out under standard conditions well known to those skilled in the art, for example at a source of palladium (eg palladium tetrakistriphenylphosphine or palladium (II) acetate), optionally phosphine ligands (eg Xantphos or S-phos ), And a suitable base (such as cesium carbonate or triethylamine).

因此具有化學式(II)之化合物在具有化學式(I)之化合物的製備中作為中間體係有用的，並且提供了另一個實施方式。 Therefore , the compound of formula (II) is useful as an intermediate system in the preparation of the compound of formula (I) , and provides another embodiment.

在一個實施方式中，提供了具有具有化學式(II)之化合物或其鹽，其中：R 1 係四氫哌喃-3-基或3,3-二甲基四氫哌喃-4-基；R 2 係甲基或氫；R 3 係氫或氟；並且X係離去基團。在一個實施方式中，X係碘、溴、或氯原子或三氟甲磺酸鹽基團。在一個實施方式中，X係溴原子。 In one embodiment, there is provided a compound of formula (II) or a salt thereof, wherein: R 1 is tetrahydropiperan-3-yl or 3,3-dimethyltetrahydropiperan-4-yl; R 2 is methyl or hydrogen; R 3 is hydrogen or fluorine; and X is a leaving group. In one embodiment, X is an iodine, bromine, or chlorine atom or a triflate group. In one embodiment, X is a bromine atom.

在一個實施方式中，提供了具有具有化學式(II)之化合物或其鹽，其中：R 1 係四氫哌喃-3-基；R 2 係甲基或氫；R 3 係氫或氟；並且X係離去基團。在一個實施方式中，X係碘、溴、或氯原子或三氟甲磺酸鹽基團。在一個實施方式中，X係溴原子。 In one embodiment, there is provided a compound of formula (II) or a salt thereof, wherein: R 1 is tetrahydropiperan-3-yl; R 2 is methyl or hydrogen; R 3 is hydrogen or fluorine; and X is a leaving group. In one embodiment, X is an iodine, bromine, or chlorine atom or a triflate group. In one embodiment, X is a bromine atom.

因此具有化學式(II)之化合物在具有化學式(I)之化合物的製備中作為中間體係有用的，並且提供了另一個實施方式。 Therefore , the compound of formula (II) is useful as an intermediate system in the preparation of the compound of formula (I) , and provides another embodiment.

在一個實施方式中，提供了具有具有化學式(II)之化合物或其鹽，其中：R 1 係3,3-二甲基四氫哌喃-4-基；R 2 係甲基或氫；R 3 係氫或氟；並且X係離去基團。在一個實施方式中，X係碘、溴、或氯原子或三氟甲磺酸鹽基團。在一個實施方式中，X係溴原子。 In one embodiment, a compound of formula (II) or a salt thereof is provided, wherein: R 1 is 3,3-dimethyltetrahydropiperan-4-yl; R 2 is methyl or hydrogen; R 3 is hydrogen or fluorine; and X is a leaving group. In one embodiment, X is an iodine, bromine, or chlorine atom or a triflate group. In one embodiment, X is a bromine atom.

因此具有化學式(II)之化合物在具有化學式(I)之化合物的製備中作為中間體係有用的，並且提供了另一個實施方式。 Therefore , the compound of formula (II) is useful as an intermediate system in the preparation of the compound of formula (I) , and provides another embodiment.

在一個實施方式中，提供了具有具有化學式(II)之化合物或其鹽，其中：R 1 係3,3-二甲基四氫哌喃-4-基；R 2 係甲基；R 3 係氫；並且X係離去基團。在一個實施方式中，X係碘、溴、或氯原子或三氟甲磺酸鹽基團。在一個實施方式中，X係溴原子。 In one embodiment, a compound of formula (II) or a salt thereof is provided, wherein: R 1 is 3,3-dimethyltetrahydropiperan-4-yl; R 2 is methyl; R 3 is Hydrogen; and X is a leaving group. In one embodiment, X is an iodine, bromine, or chlorine atom or a triflate group. In one embodiment, X is a bromine atom.

在具有化學式(II)之化合物或其鹽被提及的任何實施方式中，要理解的是此類鹽不必是藥學上可接受的鹽。具有化學式(II)之化合物的適合的鹽例如是酸加成鹽。具有化學式(II)之化合物的酸加成鹽可以藉由在技術人員已知的條件下使該化合物與適合的無機酸或有機酸接觸來形成。酸加成鹽例如可以使用選自鹽酸、氫溴酸、硫酸和磷酸的無機酸來形成。酸加成鹽還可以使用選自以下各項的有機酸來形成：三氟乙酸、檸檬酸、馬來酸、草酸、乙酸、甲酸、苯甲酸、富馬酸、琥珀酸、酒石酸、乳酸、丙酮酸、甲磺酸、苯磺酸以及對甲苯磺酸。 In any embodiment where a compound of formula (II) or a salt thereof is mentioned, it is understood that such salts need not be pharmaceutically acceptable salts. Suitable salts of compounds of formula (II) are, for example, acid addition salts. The acid addition salt of the compound of formula (II) can be formed by contacting the compound with a suitable inorganic acid or organic acid under conditions known to the skilled person. The acid addition salt can be formed using, for example, an inorganic acid selected from hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid. Acid addition salts can also be formed using organic acids selected from the group consisting of trifluoroacetic acid, citric acid, maleic acid, oxalic acid, acetic acid, formic acid, benzoic acid, fumaric acid, succinic acid, tartaric acid, lactic acid, acetone Acid, methanesulfonic acid, benzenesulfonic acid and p-toluenesulfonic acid.

因此，在一個實施方式中，提供了具有化學式(II)之化合物或其鹽，其中該鹽係鹽酸鹽、氫溴酸鹽、硫酸鹽、磷酸鹽、三氟乙酸鹽、檸檬酸鹽、馬來酸鹽、草酸鹽、乙酸鹽、甲酸鹽、苯甲酸鹽、富馬酸鹽、琥珀酸鹽、酒石酸鹽、乳酸鹽、丙酮酸鹽、甲磺酸鹽、苯磺酸鹽或對甲苯磺酸鹽。 Therefore, in one embodiment, there is provided a compound of formula (II) or a salt thereof, wherein the salt is hydrochloride, hydrobromide, sulfate, phosphate, trifluoroacetate, citrate, horse Salt, oxalate, acetate, formate, benzoate, fumarate, succinate, tartrate, lactate, pyruvate, methanesulfonate, benzenesulfonate or Tosylate.

在一個實施方式中，提供了具有化學式(II)之化合物或其鹽，其中該化合物選自：8-溴-3-甲基-1-[(3R)-氧雜環己烷-3-基)咪唑并[5,4-c]喹啉-2-酮；8-溴-7-氟-3-甲基-1-[(3S)-氧雜環己烷-3-基)咪唑并[5,4-c]喹啉-2-酮；8-溴-7-氟-3-甲基-1-[(3R)-氧雜環己烷-3-基)咪唑并[5,4-c]喹啉-2-酮； 外消旋-8-溴-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮；外消旋-8-溴-7-氟-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮；外消旋-8-溴-1-(3,3-二甲基四氫-2H-哌喃-4-基)-3-甲基-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮；8-溴-3-甲基-1-[(3R)-氧雜環己烷-3-基)咪唑并[5,4-c]喹啉-2-酮；8-溴-7-氟-3-甲基-1-[(3S)-氧雜環己烷-3-基)咪唑并[5,4-c]喹啉-2-酮；8-溴-7-氟-3-甲基-1-[(3R)-氧雜環己烷-3-基)咪唑并[5,4-c]喹啉-2-酮；外消旋-8-溴-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮；以及外消旋-8-溴-7-氟-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮。 In one embodiment, a compound of formula (II) or a salt thereof is provided, wherein the compound is selected from the group consisting of: 8-bromo-3-methyl-1-[(3 R ) -oxetan-3- Group) imidazo [5,4-c] quinolin-2-one; 8-bromo-7-fluoro-3-methyl-1-[(3 S ) -oxetan-3-yl) imidazole [5,4-c] quinolin-2-one; 8-bromo-7-fluoro-3-methyl-1-[(3 R ) -oxetan-3-yl) imidazo [5 , 4-c] quinolin-2-one; racemic-8-bromo-3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5- c] Quinolin-2 (3H) -one; racemic-8-bromo-7-fluoro-3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [ 4,5-c] quinoline-2 (3H) -one; racemic-8-bromo-1- (3,3-dimethyltetrahydro-2H-piperan-4-yl) -3-methyl Yl-1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one; 8-bromo-3-methyl-1-[(3 R ) -oxetanyl- 3-yl) imidazo [5,4-c] quinolin-2-one; 8-bromo-7-fluoro-3-methyl-1-[(3 S ) -oxetan-3-yl ) Imidazo [5,4-c] quinolin-2-one; 8-bromo-7-fluoro-3-methyl-1-[(3 R ) -oxetan-3-yl) imidazo [5,4-c] quinolin-2-one; racemic-8-bromo-3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4, 5-c] quinoline-2 (3H) -one; and racemic -8-Bromo-7-fluoro-3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5-c] quinolin-2 (3H) -one .

具有化學式(III)之化合物可以藉由類似於實例部分示出的那些方法來製備。 Compounds of formula (III) can be prepared by methods similar to those shown in the example section.

在一個實施方式中，提供了在實驗部分中所描述的新穎的中間體中的任何一個。 In one embodiment, any one of the novel intermediates described in the experimental section is provided.

作為其ATM激酶抑制活性的結果，具有化學式(I) 之化合物、及其藥學上可接受的鹽預期在療法中(例如在至少部分由ATM激酶介導的疾病或醫學病狀，包括癌症的治療中)係有用的。 As a result of its ATM kinase inhibitory activity, the compound of formula (I) and its pharmaceutically acceptable salts are expected to be used in therapy (eg in the treatment of diseases or medical conditions that are at least partially mediated by ATM kinase, including cancer Medium) is useful.

在提及“癌症”的情況下，這包括非轉移性癌症和轉移性癌症兩者，使得治療癌症涉及治療原發性腫瘤和腫瘤轉移兩者。 Where "cancer" is mentioned, this includes both non-metastatic and metastatic cancers, so that treating cancer involves treating both primary tumors and tumor metastases.

“ATM激酶抑制活性”係指作為對具有化學式(I)之化合物或其藥學上可接受的鹽的存在下的直接或間接響應，ATM激酶的活性相對於在不存在具有化學式(I)之化合物或其藥學上可接受的鹽下ATM激酶的活性降低。此類活性的降低可以由於具有化學式(I)之化合物或其藥學上可接受的鹽與ATM激酶的直接相互作用，或由於具有化學式(I)之化合物或其藥學上可接受的鹽與一種或多種反過來影響ATM激酶活性的其他因素相互作用。例如，具有化學式(I)之化合物或其藥學上可接受的鹽可以藉由直接與ATM激酶結合、藉由(直接或間接)引起另一因素降低ATM激酶活性、或藉由(直接或間接)降低存在於細胞或有機體中的ATM激酶的量來降低ATM激酶。 "ATM kinase inhibitory activity" means as a direct or indirect response to the presence of a compound of formula (I) or a pharmaceutically acceptable salt thereof, the activity of ATM kinase relative to the absence of a compound of formula (I) Or the pharmacologically acceptable salt of ATM kinase activity is reduced. Such reduction in activity may be due to the direct interaction of the compound of formula (I) or a pharmaceutically acceptable salt thereof with ATM kinase, or the compound of formula (I) or a pharmaceutically acceptable salt thereof with one or A variety of other factors that in turn affect ATM kinase activity interact. For example, a compound of formula (I) or a pharmaceutically acceptable salt thereof can be combined with ATM kinase directly, by (directly or indirectly) causing another factor to reduce ATM kinase activity, or by (directly or indirectly) ATM kinase is reduced by reducing the amount of ATM kinase present in the cell or organism.

術語“療法”旨在具有其正常的含義：處理疾病，以便完全或部分緩解其症狀的一種、一些或全部，或以便針對潛在病理進行糾正或補償。術語“療法”還包括“預防”，除非有相反的具體指示。術語“治療的”和“治療地”應以相應的方式被解釋。 The term "therapy" is intended to have its normal meaning: to treat a disease in order to completely or partially relieve one, some, or all of its symptoms, or to correct or compensate for the underlying pathology. The term "therapy" also includes "prevention" unless specifically indicated to the contrary. The terms "therapeutic" and "therapeutic" should be interpreted in a corresponding manner.

術語“預防”旨在具有其正常的含義，並包括防止 疾病發展的初級預防和繼發性預防，其中該疾病已經發展並且患者被暫時或永久保護對抗疾病的加重或惡化或者對抗與疾病相關的新症狀的發展。 The term "prevention" is intended to have its normal meaning and includes primary prevention and secondary prevention to prevent the development of the disease, where the disease has developed and the patient is temporarily or permanently protected against the aggravation or worsening of the disease or against the disease-related The development of new symptoms.

術語“治療”(treatment)與“療法”(therapy)同義地使用。類似地，術語“治療”(treat)可視為“施加療法”(applying therapy)，其中“療法”(therapy)係如本文所定義的。 The term "treatment" is used synonymously with "therapy". Similarly, the term "treat" can be regarded as "applying therapy", where "therapy" is as defined herein.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在療法中使用。 In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in therapy.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽在生產藥物中之用途。 In one embodiment, there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament.

在一個實施方式中，提供了具有化學式(I)之化合物，或其藥學上可接受的鹽，用於在治療由ATM激酶介導的疾病中使用。 In one embodiment, a compound of formula (I) , or a pharmaceutically acceptable salt thereof, is provided for use in the treatment of diseases mediated by ATM kinase.

在一個實施方式中，提供了具有化學式(I)之化合物，或其藥學上可接受的鹽，用於在治療由ATM激酶介導的疾病中使用，其中由ATM激酶介導的所述疾病係癌症。 In one embodiment, there is provided a compound of formula (I) , or a pharmaceutically acceptable salt thereof, for use in the treatment of diseases mediated by ATM kinase, wherein the diseases mediated by ATM kinase are cancer.

在一個實施方式中，提供了具有化學式(I)之化合物，或其藥學上可接受的鹽，用於在治療由ATM激酶介導的疾病中使用，其中由ATM激酶介導的所述疾病係結腸直腸癌、惡性膠質瘤、胃癌、卵巢癌、彌漫性大B細胞淋巴瘤、慢性淋巴球性白血病、急性髓性白血病、頭頸部鱗狀細胞癌、乳癌、肝細胞癌、小細胞肺癌或非小細胞肺癌。 In one embodiment, there is provided a compound of formula (I) , or a pharmaceutically acceptable salt thereof, for use in the treatment of diseases mediated by ATM kinase, wherein the diseases mediated by ATM kinase are Colorectal cancer, malignant glioma, gastric cancer, ovarian cancer, diffuse large B-cell lymphoma, chronic lymphocytic leukemia, acute myeloid leukemia, head and neck squamous cell carcinoma, breast cancer, hepatocellular carcinoma, small cell lung cancer or non- Small Cell Lung Cancer.

在一個實施方式中，提供了具有化學式(I)之化合物，或其藥學上可接受的鹽，用於在治療由ATM激酶介導的疾病中使用，其中由ATM激酶介導的所述疾病係結腸直腸癌。 In one embodiment, there is provided a compound of formula (I) , or a pharmaceutically acceptable salt thereof, for use in the treatment of diseases mediated by ATM kinase, wherein the diseases mediated by ATM kinase are Colorectal cancer.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用。 In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of cancer.

在一個實施方式中，提供了具有化學式(I)之化合物，或其藥學上可接受的鹽，用於在以下疾病的治療中使用：結腸直腸癌、惡性膠質瘤、胃癌、卵巢癌、彌漫性大B細胞淋巴瘤、慢性淋巴球性白血病、急性髓性白血病、頭頸部鱗狀細胞癌、乳癌、肝細胞癌、小細胞肺癌或非小細胞肺癌。 In one embodiment, a compound of formula (I) , or a pharmaceutically acceptable salt thereof, is provided for use in the treatment of the following diseases: colorectal cancer, malignant glioma, gastric cancer, ovarian cancer, diffuse Large B-cell lymphoma, chronic lymphocytic leukemia, acute myeloid leukemia, head and neck squamous cell carcinoma, breast cancer, hepatocellular carcinoma, small cell lung cancer or non-small cell lung cancer.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在結腸直腸癌的治療中使用。 In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of colorectal cancer.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在杭丁頓氏症(Huntingdon’s disease)的治療中使用。 In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of Huntingdon's disease.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於用作神經保護劑。 In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use as a neuroprotective agent.

“神經保護劑”係指保存神經元結構和/或功能的試劑。 "Neuroprotectant" refers to an agent that preserves the structure and / or function of neurons.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽在生產用於治療由ATM激酶介導的疾病的藥物中之用途。 In one embodiment, there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating a disease mediated by ATM kinase.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽在生產用於治療由ATM激酶介導的疾病的藥物中之用途，其中由ATM激酶介導的所述疾病係癌症。 In one embodiment, there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of diseases mediated by ATM kinase, wherein said mediated by ATM kinase The disease is cancer.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽在生產用於治療由ATM激酶介導的疾病的藥物中的用途，其中由ATM激酶介導的所述疾病係結腸直腸癌、惡性膠質瘤、胃癌、卵巢癌、彌漫性大B細胞淋巴瘤、慢性淋巴球性白血病、急性髓性白血病、頭頸部鱗狀細胞癌、乳癌、肝細胞癌、小細胞肺癌以及非小細胞肺癌。 In one embodiment, there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of diseases mediated by ATM kinase, wherein said mediated by ATM kinase Diseases include colorectal cancer, malignant glioma, gastric cancer, ovarian cancer, diffuse large B-cell lymphoma, chronic lymphocytic leukemia, acute myeloid leukemia, head and neck squamous cell carcinoma, breast cancer, hepatocellular carcinoma, small cell lung cancer And non-small cell lung cancer.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽在生產用於治療由ATM激酶介導的疾病的藥物中的用途，其中由ATM激酶介導的所述疾病係結腸直腸癌。 In one embodiment, there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of diseases mediated by ATM kinase, wherein said mediated by ATM kinase The disease is colorectal cancer.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽在生產用於治療癌症的藥物中之用途。 In one embodiment, there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating cancer.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽在生產用於治療以下疾病的藥物中的用途：結腸直腸癌、惡性膠質瘤、胃癌、卵巢癌、彌漫性大B細胞淋巴瘤、慢性淋巴球性白血病、急性髓性白血病、頭頸部鱗狀細胞癌、乳癌、肝細胞癌、小細胞肺癌或非小細胞肺癌。 In one embodiment, there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of the following diseases: colorectal cancer, malignant glioma, gastric cancer, ovarian cancer, diffuse Large B-cell lymphoma, chronic lymphocytic leukemia, acute myeloid leukemia, squamous cell carcinoma of the head and neck, breast cancer, hepatocellular carcinoma, small cell lung cancer or non-small cell lung cancer.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽在生產用於治療結腸直腸癌的藥物中之用途。 In one embodiment, the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating colorectal cancer is provided.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽在生產用於治療杭丁頓氏症的藥物中之用途。 In one embodiment, there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating Huntington's disease.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽在生產用作神經保護劑的藥物中之用途。 In one embodiment, the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for use as a neuroprotective agent is provided.

在一個實施方式中，提供了在需要這種治療的溫血動物中用於治療其中ATM激酶的抑制係有益的疾病之方法，該方法包括向所述溫血動物給予治療有效量的具有化學式(I)之化合物或其藥學上可接受的鹽。 In one embodiment, a method for treating a disease in which inhibition of ATM kinase is beneficial in a warm-blooded animal in need of such treatment is provided, the method comprising administering to the warm-blooded animal a therapeutically effective amount of a chemical formula ( The compound of I) or a pharmaceutically acceptable salt thereof.

術語“治療有效量”係指如在本文的任何實施方式中所描述的具有化學式(I)之化合物的量，該量有效地在受試者中提供“療法”，或在受試者中有效地“治療”疾病或失調。在癌症的情況下，如在以上“療法”、“治療”和“預防”的定義中所述的，治療有效量可以在受試者中引起任何可觀察的或可測量的變化。例如，該有效量可以降低癌或腫瘤細胞的數量；降低總體腫瘤大小；抑制或停止腫瘤細胞浸潤至外周器官，例如包括軟組織和骨；抑制並停止腫瘤轉移；抑制並停止腫瘤生長；在某種程度上緩解與癌症相關的症狀中的一種或多種；降低發病率和死亡率；提高生命品質；或該等作用的組合。有效量可以是足以減少響應於ATM激酶活性的抑制的疾病的症狀的量。對於癌症療法，例如可以藉由評估存活期、疾病進展時間(TTP)、應答率(RR)、響應期、和/或生命品質來測定體內療效。如由熟習該項 技術者所認可的，有效量可以取決於給予途徑、賦形劑的使用、以及與其他藥劑共同使用而改變。例如，在使用聯合療法的情況下，在動物患者中，對於治療靶向的失調，當聯合時，本說明書中描述的具有化學式(I)之化合物或藥學上可接受的鹽的量和其他一種或多種藥學上有活性的藥劑的量係共同有效的。在該背景下，如果它們在組合時足以降低如以上所述的響應於ATM活性抑制的疾病的症狀，組合的量係“治療有效量”的。典型地，熟習該項技術者可以藉由例如從針對具有化學式(I)之化合物或其藥學上可接受的鹽的、本說明書中所描述的劑量範圍開始，以及從其他一種或多種藥學上有活性的化合物的一個或多個批准的或另外公開的劑量範圍開始，來確定此類量。 The term "therapeutically effective amount" refers to an amount of a compound of formula (I) as described in any of the embodiments herein, which amount is effective to provide a "therapy" in a subject, or to be effective in a subject "Treat" a disease or disorder. In the case of cancer, as described in the definitions of "therapy", "treatment" and "prevention" above, a therapeutically effective amount can cause any observable or measurable change in the subject. For example, the effective amount can reduce the number of cancer or tumor cells; reduce the overall tumor size; inhibit or stop tumor cell infiltration into peripheral organs, including, for example, soft tissue and bone; inhibit and stop tumor metastasis; inhibit and stop tumor growth; in a certain way To alleviate one or more of the symptoms associated with cancer; reduce morbidity and mortality; improve quality of life; or a combination of these effects. The effective amount may be an amount sufficient to reduce the symptoms of the disease in response to inhibition of ATM kinase activity. For cancer therapy, for example, in vivo efficacy can be determined by assessing survival time, disease progression time (TTP), response rate (RR), response period, and / or quality of life. As recognized by those skilled in the art, the effective amount may vary depending on the route of administration, use of excipients, and co-use with other agents. For example, in the case of using combination therapy, in animal patients, for the treatment of targeted disorders, when combined, the amount of the compound of formula (I) or a pharmaceutically acceptable salt described in this specification and the other one The amount of multiple pharmacologically active agents is effective together. In this context, if they are combined enough to reduce the symptoms of a disease as described above in response to inhibition of ATM activity, the combined amount is a "therapeutically effective amount." Typically, those skilled in the art can start by, for example, starting from the dosage range described in this specification for a compound of formula (I) or a pharmaceutically acceptable salt thereof, and from one or more other pharmacologically available One or more approved or otherwise disclosed dosage ranges of the active compound begin to determine such amounts.

“溫血動物”包括例如人。 "Warm-blooded animals" include, for example, humans.

在一個實施方式中，提供了在需要這種治療的溫血動物中用於治療其中ATM激酶的抑制係有益的疾病之方法，該方法包括向所述溫血動物給予治療有效量的具有化學式(I)之化合物或其藥學上可接受的鹽，並且其中ATM激酶的抑制係有益的所述疾病係癌症。 In one embodiment, a method for treating a disease in which inhibition of ATM kinase is beneficial in a warm-blooded animal in need of such treatment is provided, the method comprising administering to the warm-blooded animal a therapeutically effective amount of a chemical formula ( The compound of I) or a pharmaceutically acceptable salt thereof, and the disease in which the inhibition of ATM kinase is beneficial is cancer.

在一個實施方式中，提供了在需要這種治療的溫血動物中用於治療其中ATM激酶的抑制係有益的疾病之方法，該方法包括向所述溫血動物給予治療有效量的具有化學式(I)之化合物或其藥學上可接受的鹽，並且其中ATM激酶的抑制係有益的所述疾病係結腸直腸癌、惡性膠質瘤、胃癌、卵巢癌、彌漫性大B細胞淋巴瘤、慢性淋巴球性白血病、急性髓性白血病、頭頸部鱗狀 細胞癌、乳癌、肝細胞癌、小細胞肺癌或非小細胞肺癌。 In one embodiment, a method for treating a disease in which inhibition of ATM kinase is beneficial in a warm-blooded animal in need of such treatment is provided, the method comprising administering to the warm-blooded animal a therapeutically effective amount of a chemical formula ( I) A compound or a pharmaceutically acceptable salt thereof, and wherein the inhibition of ATM kinase is beneficial The diseases are colorectal cancer, malignant glioma, gastric cancer, ovarian cancer, diffuse large B-cell lymphoma, chronic lymphocytic Leukemia, acute myeloid leukemia, head and neck squamous cell carcinoma, breast cancer, hepatocellular carcinoma, small cell lung cancer or non-small cell lung cancer.

在一個實施方式中，提供了在需要這種治療的溫血動物中用於治療其中ATM激酶的抑制係有益的疾病之方法，該方法包括向所述溫血動物給予治療有效量的具有化學式(I)之化合物或其藥學上可接受的鹽，並且其中ATM激酶的抑制係有益的所述疾病係結腸直腸癌。 In one embodiment, a method for treating a disease in which inhibition of ATM kinase is beneficial in a warm-blooded animal in need of such treatment is provided, the method comprising administering to the warm-blooded animal a therapeutically effective amount of a chemical formula ( The compound of I) or a pharmaceutically acceptable salt thereof, and the disease in which the inhibition of ATM kinase is beneficial is colorectal cancer.

在一個實施方式中，提供了在需要這種治療的溫血動物中用於治療其中ATM激酶的抑制係有益的疾病之方法，該方法包括向所述溫血動物給予治療有效量的具有化學式(I)之化合物或其藥學上可接受的鹽，並且其中ATM激酶的抑制係有益的所述疾病係杭丁頓氏症。 In one embodiment, a method for treating a disease in which inhibition of ATM kinase is beneficial in a warm-blooded animal in need of such treatment is provided, the method comprising administering to the warm-blooded animal a therapeutically effective amount of a chemical formula ( The compound of I) or a pharmaceutically acceptable salt thereof, and the disease in which the inhibition of ATM kinase is beneficial is Huntington's disease.

在一個實施方式中，提供了在需要此類治療的溫血動物中用於治療癌症之方法，該方法包括向所述溫血動物給予治療有效量的具有化學式(I)之化合物或其藥學上可接受的鹽。 In one embodiment, a method for treating cancer in a warm-blooded animal in need of such treatment is provided, the method comprising administering to the warm-blooded animal a therapeutically effective amount of a compound of formula (I) or a pharmaceutical thereof Acceptable salt.

在一個實施方式中，提供了在需要此類治療的溫血動物中用於治療以下疾病之方法：結腸直腸癌、惡性膠質瘤、胃癌、卵巢癌、彌漫性大B細胞淋巴瘤、慢性淋巴球性白血病、急性髓性白血病、頭頸部鱗狀細胞癌、乳癌、肝細胞癌、小細胞肺癌或非小細胞肺癌，該方法包括向所述溫血動物給予治療有效量的具有化學式(I)之化合物或其藥學上可接受的鹽。 In one embodiment, a method for treating the following diseases in warm-blooded animals in need of such treatment is provided: colorectal cancer, malignant glioma, gastric cancer, ovarian cancer, diffuse large B-cell lymphoma, chronic lymphocytic Leukemia, acute myeloid leukemia, squamous cell carcinoma of the head and neck, breast cancer, hepatocellular carcinoma, small cell lung cancer or non-small cell lung cancer, the method includes administering to the warm-blooded animal a therapeutically effective amount of formula (I) Compound or a pharmaceutically acceptable salt thereof.

在一個實施方式中，提供了在需要此類治療的溫血動物中用於治療結腸直腸癌之方法，該方法包括向所述溫血動物 給予治療有效量的具有化學式(I)之化合物或其藥學上可接受的鹽。 In one embodiment, a method for treating colorectal cancer in a warm-blooded animal in need of such treatment is provided, the method comprising administering to the warm-blooded animal a therapeutically effective amount of a compound of formula (I) or Pharmaceutically acceptable salts.

在一個實施方式中，提供了在需要此類治療的溫血動物中用於治療杭丁頓氏症之方法，該方法包括向所述溫血動物給予治療有效量的具有化學式(I)之化合物或其藥學上可接受的鹽。 In one embodiment, a method for treating Huntington's disease in a warm-blooded animal in need of such treatment is provided, the method comprising administering to the warm-blooded animal a therapeutically effective amount of a compound of formula (I) Or a pharmaceutically acceptable salt thereof.

在一個實施方式中，提供了在需要此類治療的溫血動物中用於影響神經保護作用之方法，該方法包括向所述溫血動物給予治療有效量的具有化學式(I)之化合物或其藥學上可接受的鹽。 In one embodiment, a method for affecting neuroprotection in a warm-blooded animal in need of such treatment is provided, the method comprising administering to the warm-blooded animal a therapeutically effective amount of a compound of formula (I) or Pharmaceutically acceptable salts.

在一個實施方式中，提供了在需要此類治療的溫血動物中用於治療癌症之方法，該方法包括向所述溫血動物給予治療有效量的具有化學式(I)之化合物或其藥學上可接受的鹽。在一個實施方式中，所述癌症選自結腸直腸癌、惡性膠質瘤、胃癌、卵巢癌、彌漫性大B細胞淋巴瘤、慢性淋巴球性白血病、急性髓性白血病、頭頸部鱗狀細胞癌、乳癌、肝細胞癌、小細胞肺癌以及非小細胞肺癌。在一個實施方式中，所述癌症選自結腸直腸癌、惡性膠質瘤、胃癌、卵巢癌、彌漫性大B細胞淋巴瘤、慢性淋巴球性白血病、頭頸部鱗狀細胞癌以及肺癌。在一個實施方式中，所述癌症係結腸直腸癌。 In one embodiment, a method for treating cancer in a warm-blooded animal in need of such treatment is provided, the method comprising administering to the warm-blooded animal a therapeutically effective amount of a compound of formula (I) or a pharmaceutical thereof Acceptable salt. In one embodiment, the cancer is selected from colorectal cancer, malignant glioma, gastric cancer, ovarian cancer, diffuse large B-cell lymphoma, chronic lymphocytic leukemia, acute myeloid leukemia, squamous cell carcinoma of the head and neck, Breast cancer, hepatocellular carcinoma, small cell lung cancer and non-small cell lung cancer. In one embodiment, the cancer is selected from colorectal cancer, malignant glioma, gastric cancer, ovarian cancer, diffuse large B-cell lymphoma, chronic lymphocytic leukemia, squamous cell carcinoma of the head and neck, and lung cancer. In one embodiment, the cancer is colorectal cancer.

在癌症以一般意義被提及的任何實施方式中，所述癌症可以選自結腸直腸癌、惡性膠質瘤、胃癌、卵巢癌、彌漫性大B細胞淋巴瘤、慢性淋巴球性白血病、急性髓性白血病、頭頸部 In any embodiment where cancer is mentioned in a general sense, the cancer may be selected from colorectal cancer, malignant glioma, gastric cancer, ovarian cancer, diffuse large B-cell lymphoma, chronic lymphocytic leukemia, acute myeloid Leukemia, head and neck

鱗狀細胞癌、乳癌、肝細胞癌、小細胞肺癌以及非小細胞肺癌。 Squamous cell carcinoma, breast cancer, hepatocellular carcinoma, small cell lung cancer and non-small cell lung cancer.

在癌症以一般意義被提及的任何實施方式中，可以採用以下實施方式：在一個實施方式中，該癌症係結腸直腸癌。 In any embodiment where the cancer is mentioned in a general sense, the following embodiment may be adopted: In one embodiment, the cancer is colorectal cancer.

在一個實施方式中，該癌症係惡性膠質瘤。 In one embodiment, the cancer is malignant glioma.

在一個實施方式中，該癌症係胃癌。 In one embodiment, the cancer is gastric cancer.

在一個實施方式中，該癌症係食道癌。 In one embodiment, the cancer is esophageal cancer.

在一個實施方式中，該癌症係卵巢癌。 In one embodiment, the cancer is ovarian cancer.

在一個實施方式中，該癌症係子宮內膜癌。 In one embodiment, the cancer is endometrial cancer.

在一個實施方式中，該癌症係子宮頸癌。 In one embodiment, the cancer is cervical cancer.

在一個實施方式中，該癌症係彌漫性大B細胞淋巴瘤。 In one embodiment, the cancer is diffuse large B-cell lymphoma.

在一個實施方式中，該癌症係慢性淋巴球性白血病。 In one embodiment, the cancer is chronic lymphocytic leukemia.

在一個實施方式中，該癌症係急性髓性白血病。 In one embodiment, the cancer is acute myeloid leukemia.

在一個實施方式中，該癌症係頭頸部鱗狀細胞癌。 In one embodiment, the cancer is head and neck squamous cell carcinoma.

在一個實施方式中，該癌症係乳癌。在一個實施方式中，該癌症係三陰性乳癌。 In one embodiment, the cancer is breast cancer. In one embodiment, the cancer is triple negative breast cancer.

“三陰性乳癌”係針對***受體、孕酮受體和Her2/neu沒有測試為陽性的任何乳癌。用於測定關於該等受體中的每一種的陽性測試的測試方法係本領域熟知的。 "Triple negative breast cancer" refers to any breast cancer that does not test positive for estrogen receptor, progesterone receptor and Her2 / neu. Test methods for determining a positive test for each of these receptors are well known in the art.

在一個實施方式中，該癌症係肝細胞癌。 In one embodiment, the cancer is hepatocellular carcinoma.

在一個實施方式中，該癌症係肺癌。在一個實施方式中，該肺癌係小細胞肺癌。在一個實施方式中，該肺癌係非小細胞肺癌。 In one embodiment, the cancer is lung cancer. In one embodiment, the lung cancer is small cell lung cancer. In one embodiment, the lung cancer is non-small cell lung cancer.

在一個實施方式中，該癌症係轉移性癌症。在一個實施方式中，該轉移性癌症包括中樞神經系統的轉移。在一個實施方式中，該中樞神經系統的轉移包括腦轉移。在一個實施方式中，該中樞神經系統的轉移包括柔腦膜轉移。 In one embodiment, the cancer is metastatic cancer. In one embodiment, the metastatic cancer includes metastases from the central nervous system. In one embodiment, the central nervous system metastasis includes brain metastasis. In one embodiment, the central nervous system metastasis includes pia mater.

當癌症擴散到腦膜(覆蓋腦和脊髓的組織層)時，“柔腦膜轉移”發生。轉移可以藉由血液擴散至腦膜，或它們可以從腦轉移開始行進，該腦轉移由流經腦膜的腦脊髓液(CSF)運載。在一個實施方式中，該癌症係非轉移性癌症。 When cancer spreads to the meninges (the tissue layer covering the brain and spinal cord), "flexible meningeal metastases" occur. Metastases can spread to the meninges by blood, or they can proceed from brain metastases, which are carried by cerebrospinal fluid (CSF) that flows through the meninges. In one embodiment, the cancer is a non-metastatic cancer.

在本說明書中所描述的抗癌治療可以作為單一療法係有用的，或者除了給予具有化學式(I)之化合物以外，還可以包括常規手術、放射療法或化學療法；或此類另外的療法的組合。 這種常規手術、放射療法或化學療法可以與具有化學式(I)之化合物同時地、順序地或分別地給予，以進行治療。 The anti-cancer treatment described in this specification may be useful as a monotherapy, or may include conventional surgery, radiation therapy, or chemotherapy in addition to the compound of formula (I) ; or a combination of such additional therapies . Such conventional surgery, radiation therapy or chemotherapy can be administered simultaneously, sequentially or separately with the compound of formula (I) for treatment.

放射療法可以包括以下類別的療法中的一種或多種：i.使用電磁輻射的外部放射療法，和使用電磁輻射的術中放射療法；ii.內部放射療法或近距離放射療法；包括間質性放射療法或腔內放射療法；或iii.全身放射療法，包括但不限於碘131和鍶89。 Radiation therapy may include one or more of the following categories of therapy: i. External radiation therapy using electromagnetic radiation, and intraoperative radiotherapy using electromagnetic radiation; ii. Internal radiation therapy or brachytherapy; including interstitial radiation therapy Or intracavitary radiation therapy; or iii. Systemic radiation therapy, including but not limited to iodine 131 and strontium 89.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用，其中具有化學式(I)之化合物或其藥學上可接受的鹽與放射療法被組合給予。在一個實施方式中，該放射療法選自列於以上點(i)-(iii)下的一種或多種類別的放射療法。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is Radiation therapy is given in combination. In one embodiment, the radiotherapy is selected from one or more categories of radiotherapy listed under points (i)-(iii) above.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在治療以下疾病中使用：惡性膠質瘤、肺癌(例如小細胞肺癌或非小細胞肺癌)、乳癌(例如三陰性乳癌)、頭頸部鱗狀細胞癌、食道癌、子宮頸癌或子宮內膜癌，其中具有化學式(I)之化合物或其藥學上可接受的鹽與放射療法被組合給予。在一個實施方式中，該放射療法選自列於以上點(i)-(iii)下的一種或多種類別的放射療法。 In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of the following diseases: malignant glioma, lung cancer (eg, small cell lung cancer or non-small cell lung cancer), Breast cancer (e.g. triple negative breast cancer), head and neck squamous cell carcinoma, esophageal cancer, cervical cancer or endometrial cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with radiation therapy. In one embodiment, the radiotherapy is selected from one or more categories of radiotherapy listed under points (i)-(iii) above.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在惡性膠質瘤的治療中使用，其中具有化學式(I)之化合物或其藥學上可接受的鹽與放射療法被組合給予。在一個實施方式中，該放射療法選自列於以上點(i)-(iii)下的一種或多種類別的放射療法。 In one embodiment, a salt with a compound of formula acceptable (I) or of a pharmaceutically acceptable for use in the treatment of malignant glioma, having acceptable compounds of formula (I) or the pharmaceutically Salt and radiation therapy are given in combination. In one embodiment, the radiotherapy is selected from one or more categories of radiotherapy listed under points (i)-(iii) above.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在轉移性癌症的治療中使用，其中具有化學式(I)之化合物或其藥學上可接受的鹽與放射療法被組合給予。在一個實施方式中，該放射療法選自列於以上點(i)-(iii)下的一種或多種類別的放射療法。 In one embodiment, a salt with a compound of formula acceptable (I) or of a pharmaceutically acceptable for use in the treatment of metastatic cancer, wherein the compound having acceptable formula (I) or the pharmaceutically Salt and radiation therapy are given in combination. In one embodiment, the radiotherapy is selected from one or more categories of radiotherapy listed under points (i)-(iii) above.

在一個實施方式中，提供了具有化學式(I)之化合 物或其藥學上可接受的鹽，用於在中樞神經系統的轉移的治療中使用，其中具有化學式(I)之化合物或其藥學上可接受的鹽與放射療法被組合給予。在一個實施方式中，該放射療法選自列於以上點(i)-(iii)下的一種或多種類別的放射療法。 In one embodiment, a salt with a compound of formula acceptable (I) or the pharmaceutically acceptable for use in the treatment of metastatic central nervous system, wherein a compound having the formula (I) or a pharmaceutically acceptable of The received salt is given in combination with radiation therapy. In one embodiment, the radiotherapy is selected from one or more categories of radiotherapy listed under points (i)-(iii) above.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在柔腦膜轉移的治療中使用，其中具有化學式(I)之化合物或其藥學上可接受的鹽與放射療法被組合給予。在一個實施方式中，該放射療法選自列於以上點(i)-(iii)下的一種或多種類別的放射療法。 In one embodiment, a salt with a compound of formula acceptable (I) or of a pharmaceutically acceptable for use in the treatment of leptomeningeal metastasis, wherein the compound having acceptable formula (I) or the pharmaceutically Salt and radiation therapy are given in combination. In one embodiment, the radiotherapy is selected from one or more categories of radiotherapy listed under points (i)-(iii) above.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與放射療法被同時地、分別地或順序地給予。在一個實施方式中，該放射療法選自列於以上點(i)-(iii)下的一種或多種類別的放射療法。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof It is administered simultaneously, separately or sequentially with radiation therapy. In one embodiment, the radiotherapy is selected from one or more categories of radiotherapy listed under points (i)-(iii) above.

在一個實施方式中，提供了在需要這種治療的溫血動物中治療癌症的方法，該方法包括向所述溫血動物給予具有化學式(I)之化合物或其藥學上可接受的鹽和放射療法，其中具有化學式(I)之化合物或其藥學上可接受的鹽、和放射療法在產生抗癌作用方面係共同有效的。在一個實施方式中，該癌症選自惡性膠質瘤、肺癌(例如小細胞肺癌或非小細胞肺癌)、乳癌(例如三陰性乳癌)、頭頸部鱗狀細胞癌、食道癌、子宮頸癌以及子宮內膜癌。在一個實施方式中，該癌症係惡性膠質瘤。在一個實施方式中，該癌症係轉移性癌症。在一個實施方式中，該轉移性癌症 包括中樞神經系統的轉移。在一個實施方式中，該中樞神經系統的轉移包括腦轉移。在一個實施方式中，該中樞神經系統的轉移包括柔腦膜轉移。在任何實施方式中，該放射療法選自列於以上點(i)-(iii)下的一種或多種類別的放射療法。 In one embodiment, a method of treating cancer in a warm-blooded animal in need of such treatment is provided, the method comprising administering to the warm-blooded animal a compound of formula (I) or a pharmaceutically acceptable salt and radiation Therapy, in which the compound of formula (I) or a pharmaceutically acceptable salt thereof, and radiation therapy are jointly effective in producing anticancer effects. In one embodiment, the cancer is selected from malignant glioma, lung cancer (eg small cell lung cancer or non-small cell lung cancer), breast cancer (eg triple negative breast cancer), head and neck squamous cell carcinoma, esophageal cancer, cervical cancer and uterus Endometrial cancer. In one embodiment, the cancer is malignant glioma. In one embodiment, the cancer is metastatic cancer. In one embodiment, the metastatic cancer includes metastases from the central nervous system. In one embodiment, the central nervous system metastasis includes brain metastasis. In one embodiment, the central nervous system metastasis includes pia mater. In any embodiment, the radiotherapy is selected from one or more categories of radiotherapy listed under points (i)-(iii) above.

在一個實施方式中，提供了在需要這種治療的溫血動物中治療癌症的方法，該方法包括向所述溫血動物給予具有化學式(I)之化合物或其藥學上可接受的鹽並且同時地、分別地或順序地給予放射療法，其中具有化學式(I)之化合物或其藥學上可接受的鹽、和放射療法在產生抗癌作用方面係共同有效的。在一個實施方式中，該癌症係惡性膠質瘤。在一個實施方式中，該癌症係轉移性癌症。在一個實施方式中，該轉移性癌症包括中樞神經系統的轉移。在一個實施方式中，該中樞神經系統的轉移包括腦轉移。在一個實施方式中，該中樞神經系統的轉移包括柔腦膜轉移。在任何實施方式中，該放射療法選自列於以上點(i)-(iii)下的一種或多種類別的放射療法。 In one embodiment, a method for treating cancer in a warm-blooded animal in need of such treatment is provided, the method comprising administering to the warm-blooded animal a compound of formula (I) or a pharmaceutically acceptable salt thereof and simultaneously Radiation therapy is administered locally, separately or sequentially, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof, and radiation therapy are jointly effective in producing anticancer effects. In one embodiment, the cancer is malignant glioma. In one embodiment, the cancer is metastatic cancer. In one embodiment, the metastatic cancer includes metastases from the central nervous system. In one embodiment, the central nervous system metastasis includes brain metastasis. In one embodiment, the central nervous system metastasis includes pia mater. In any embodiment, the radiotherapy is selected from one or more categories of radiotherapy listed under points (i)-(iii) above.

化學療法可以包括以下類別的抗腫瘤物質中的一種或多種：i.抗腫瘤劑及其組合，如DNA烷基化劑(例如順鉑(cis platin)、奧沙利鉑(oxaliplatin)、卡鉑(carboplatin)、環磷醯胺(cyclophosphamide)、氮芥樣異環磷醯胺(nitrogen mustards like ifosfamide)、苯達莫司汀(bendamustine)、美法侖(melphalan)、苯丁酸氮芥(chlorambucil)、白消安(busulphan)、替莫唑胺(temozolamide)以及亞硝基脲像卡莫司汀(carmustine))；抗代 謝物(例如吉西他濱(gemcitabine)和抗葉酸劑，如氟嘧啶(fluoropyrimidine)類，像5氟尿嘧啶和替加氟(tegafur)、雷替曲塞(raltitrexed)、甲胺蝶呤(methotrexate)、阿糖胞苷(cytosine arabinoside)、以及羥基脲)；抗腫瘤抗生素(例如蒽環類(anthracyclines)，像阿黴素(adriamycin)、博萊黴素(bleomycin)、多柔比星(doxorubicin)、脂質體多柔比星(doxorubicin)、吡柔比星(pirarubicin)、道諾黴素(daunomycin)、戊柔比星(valrubicin)表柔比星(epirubicin)、伊達比星(idarubicin)、絲裂黴素-C(mitomycin-C)、更生黴素(dactinomycin)、胺柔比星(amrubicin))以及光輝黴素(mithramycin))；抗有絲***劑(例如長春花生物鹼(vinca alkaloid)類，像長春新鹼(vincristine)、長春鹼(vinblastine)、去乙醯長春醯胺(vindesine)和長春瑞濱(vinorelbine)，以及紫杉烷類，像泰素(taxol)和多西他賽(taxotere)和保羅激酶(polokinase)抑制劑)；和拓撲異構酶抑制劑(例如表鬼臼毒素類(epipodophyllotoxins)，像依託泊苷和替尼泊苷、安吖啶(amsacrine)、伊立替康(irinotecan)、拓撲替康(topotecan)以及喜樹鹼(camptothecin))；DNA修復機制的抑制劑，如CHK激酶；DNA依賴性蛋白激酶抑制劑；聚(ADP-核糖)聚合酶的抑制劑(PARP抑制劑，包括奧拉帕尼(olaparib))；和Hsp90抑制劑，如坦螺旋黴素(tanespimycin)和瑞他黴素(retaspimycin)、ATR激酶的抑制劑(例如AZD6738)；和WEE1激酶的抑制劑(如AZD1775/MK-1775)；ii.抗血管生成劑，如抑制血管內皮生長因子的那些，例如抗血管內皮細胞生長因子抗體貝伐單抗和例如VEGF受體酪胺酸 激酶抑制劑如凡德他尼(vandetanib)(ZD6474)、索拉非尼(sorafenib)、瓦他拉尼(vatalani)(PTK787)、舒尼替尼(sunitinib)(SU11248)、阿西替尼(axitini)(AG-013736)、帕唑帕尼(pazopanib)(GW 786034)以及西地尼布(cediranib)(AZD2171)；如在國際專利申請WO 97/22596、WO 97/30035、WO 97/32856以及WO 98/13354中揭露的那些化合物；和藉由其他機理工作的化合物(例如利諾胺、整合素αvβ3功能的抑制劑和血管抑素)、或血管生成素及其受體(Tie-1和Tie-2)的抑制劑、PLGF的抑制劑、δ-樣配位基的抑制劑(DLL-4)；iii.免疫治療方法，包括例如離體和體內方法以提高患者腫瘤細胞的免疫原性，如用細胞介素如白細胞介素2、白細胞介素4或粒性白細胞-巨噬細胞集落刺激因子轉染；減少T細胞無反應性或調節性T細胞功能的方法；增強對腫瘤的T細胞應答的方法，如CTLA4(例如易普利姆瑪(ipilimumab)和曲美木單抗(tremelimumab))、B7H1、PD-1(例如BMS-936558或AMP-514)、PD-L1(例如MEDI4736)的阻斷抗體和CD137的激動劑抗體；使用轉染的免疫細胞如細胞介素轉染的樹突狀細胞的方法；使用細胞介素轉染的腫瘤細胞系的方法，對腫瘤相關抗原使用抗體，和耗盡靶細胞類型的抗體(例如未綴合的抗CD20抗體，如利妥昔單抗(Rituximab)、放射性標記的抗CD20抗體托西莫(Bexxar)和澤娃靈(Zevalin)、以及抗CD54抗體坎帕斯(Campath))的方法；使用抗獨特型抗體的方法；增強自然殺傷細胞功能的方法；和利用抗體-毒素偶聯物(例如，抗CD33抗體麥羅塔(Mylotarg))的方法；免疫毒素，如moxetumomab pasudotox；Toll樣受體7或toll 樣受體9的激動劑；iv.功效增強劑，如亞葉酸。 Chemotherapy can include one or more of the following classes of anti-tumor substances: i. Anti-tumor agents and combinations thereof, such as DNA alkylating agents (eg, cisplatin, oxaliplatin, carboplatin) (carboplatin), cyclophosphamide, nitrogen mustards like ifosfamide, bendamustine, melphalan, chlorambucil ), Busulphan, temozolamide and nitrosourea like carmustine); antimetabolites (such as gemcitabine) and antifolates such as fluoropyrimidines, Like 5-fluorouracil and tegafur, raltitrexed, methotrexate, cytosine arabinoside, and hydroxyurea); antitumor antibiotics (such as anthracyclines ( anthracyclines), like adriamycin, bleomycin, doxorubicin, doxorubicin, doxorubicin, pirarubicin, and daunorubicin ( daunomycin), valrubicin (eprubicin), epirubicin (ep irubicin), idarubicin, mitomycin-C, dactinomycin, amrubicin, and mithramycin); antimitotic agents (mithramycin); For example, vinca alkaloids, such as vincristine, vinblastine, vindesine and vinorelbine, and taxanes, like Thai (Taxol) and docetaxel (taxotere) and Paul kinase (polokinase) inhibitors; and topoisomerase inhibitors (such as epipodophyllotoxins (epipodophyllotoxins), like etoposide and teniposide, Amsacrine, irinotecan, topotecan and camptothecin); inhibitors of DNA repair mechanisms, such as CHK kinase; DNA-dependent protein kinase inhibitors; poly ( ADP-ribose) polymerase inhibitors (PARP inhibitors, including olaparib); and Hsp90 inhibitors, such as tanspiramycin (tanespimycin) and retaspimycin (retaspimycin), ATR kinase inhibition Agents (eg AZD6738); and inhibitors of WEE1 kinase (eg AZD1775 / MK-1775); ii. Angiogenesis agents, such as those that inhibit vascular endothelial growth factor, such as anti-vascular endothelial cell growth factor antibody bevacizumab and, for example, VEGF receptor tyrosine kinase inhibitors such as vandetanib (ZD6474), sora Sorafenib, vatalani (PTK787), sunitinib (SU11248), axitinib (AG-013736), pazopanib (GW 786034) and cediranib (AZD2171); as disclosed in international patent applications WO 97/22596, WO 97/30035, WO 97/32856 and WO 98/13354; and work by other mechanisms Compounds (e.g. lineolamide, integrin αvβ3 function inhibitors and angiostatin), or inhibitors of angiogenin and its receptors (Tie-1 and Tie-2), PLGF inhibitors, delta-like Ligand inhibitor (DLL-4); iii. Immunotherapy methods, including, for example, ex vivo and in vivo methods to improve the immunogenicity of the patient's tumor cells, such as the use of interleukins such as interleukin 2, interleukin 4 Or granulocyte-macrophage colony stimulating factor transfection; reduce T cell anergy or regulatory T Methods of cellular function; methods of enhancing T cell response to tumors, such as CTLA4 (eg ipilimumab and tremelimumab), B7H1, PD-1 (eg BMS-936558 or AMP -514), PD-L1 (eg MEDI4736) blocking antibody and CD137 agonist antibody; method using transfected immune cells such as cytokines transfected dendritic cells; cytokines transfected tumors Cell line method, using antibodies to tumor-associated antigens, and depleting target cell type antibodies (eg, unconjugated anti-CD20 antibodies, such as rituximab (Rituximab), radiolabeled anti-CD20 antibody tosimo ( Bexxar) and Zevalin, and the anti-CD54 antibody Campas (Campath) method; method using anti-idiotypic antibodies; methods to enhance natural killer cell function; and the use of antibody-toxin conjugates (e.g. , Anti-CD33 antibody Mylotarg (Mylotarg)); immunotoxins, such as moxetumomab pasudotox; agonists of Toll-like receptor 7 or toll-like receptor 9; iv. Efficacy enhancers, such as folinic acid.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與至少一種另外的抗腫瘤物質被組合給予。在一個實施方式中，有一種另外的抗腫瘤物質。在一個實施方式中，有兩種另外的抗腫瘤物質。在一個實施方式中，有三種或更多種另外的抗腫瘤物質。在任何實施方式中，該另外的抗腫瘤物質選自列於以上點(i)-(iv)下的一種或多種類別的抗腫瘤物質。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof It is administered in combination with at least one additional antitumor substance. In one embodiment, there is an additional anti-tumor substance. In one embodiment, there are two additional anti-tumor substances. In one embodiment, there are three or more additional anti-tumor substances. In any embodiment, the additional anti-tumor substance is selected from one or more classes of anti-tumor substances listed under points (i)-(iv) above.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與至少一種另外的抗腫瘤物質被同時地、分別地或順序地給予。在一個實施方式中，有一種另外的抗腫瘤物質。在一個實施方式中，有兩種另外的抗腫瘤物質。在一個實施方式中，有三種或更多種另外的抗腫瘤物質。在任何實施方式中，該另外的抗腫瘤物質選自列於以上點(i)-(iv)下的一種或多種類別的抗腫瘤物質。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof The at least one additional antitumor substance is administered simultaneously, separately or sequentially. In one embodiment, there is an additional anti-tumor substance. In one embodiment, there are two additional anti-tumor substances. In one embodiment, there are three or more additional anti-tumor substances. In any embodiment, the additional anti-tumor substance is selected from one or more classes of anti-tumor substances listed under points (i)-(iv) above.

在一個實施方式中，提供了在需要這種治療的溫血動物中治療癌症的方法，該方法包括向所述溫血動物給予具有化學式(I)之化合物或其藥學上可接受的鹽以及至少一種另外的抗腫瘤物質，其中具有化學式(I)之化合物或其藥學上可接受的鹽、以及另外的抗腫瘤物質的量在產生抗癌作用方面係共同有效 的。在任何實施方式中，該另外的抗腫瘤物質選自列於以上點(i)-(iv)下的一種或多種類別的抗腫瘤物質。 In one embodiment, a method for treating cancer in a warm-blooded animal in need of such treatment is provided, the method comprising administering to the warm-blooded animal a compound of formula (I) or a pharmaceutically acceptable salt thereof and at least An additional anti-tumor substance, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof, and the amount of the additional anti-tumor substance are jointly effective in producing an anti-cancer effect. In any embodiment, the additional anti-tumor substance is selected from one or more classes of anti-tumor substances listed under points (i)-(iv) above.

在一個實施方式中，提供了在需要這種治療的溫血動物中治療癌症的方法，該方法包括向所述溫血動物給予具有化學式(I)之化合物或其藥學上可接受的鹽，並且向所述溫血動物同時地、分別地或順序地給予至少一種另外的抗腫瘤物質，其中具有化學式(I)之化合物或其藥學上可接受的鹽、以及另外的抗腫瘤物質的量在產生抗癌作用方面係共同有效的。在任何實施方式中，該另外的抗腫瘤物質選自列於以上點(i)-(iv)下的一種或多種類別的抗腫瘤物質。 In one embodiment, a method of treating cancer in a warm-blooded animal in need of such treatment is provided, the method comprising administering to the warm-blooded animal a compound of formula (I) or a pharmaceutically acceptable salt thereof, and Simultaneously, separately or sequentially administer at least one additional anti-tumor substance to the warm-blooded animal, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof, and the amount of additional anti-tumor substance are produced The anti-cancer effect is common and effective. In any embodiment, the additional anti-tumor substance is selected from one or more classes of anti-tumor substances listed under points (i)-(iv) above.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽、以及至少一種抗腫瘤劑，用於在癌症的治療中使用。在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與至少一種抗腫瘤劑被組合給予。在一個實施方式中，該抗腫瘤劑選自在以上點(i)中的抗腫瘤劑的列表。 In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof, and at least one anti-tumor agent are provided for use in the treatment of cancer. In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof It is administered in combination with at least one anti-tumor agent. In one embodiment, the anti-tumor agent is selected from the list of anti-tumor agents in point (i) above.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽、以及至少一種抗腫瘤劑，用於在癌症的治療中同時、分別或順序使用。在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與至少一種抗腫瘤劑被同時地、分別地或順序地給予。在一個 實施方式中，該抗腫瘤劑選自在以上點(i)中的抗腫瘤劑的列表。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof, and at least one anti-tumor agent for simultaneous, separate or sequential use in the treatment of cancer. In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof The at least one antitumor agent is administered simultaneously, separately or sequentially. In one embodiment, the anti-tumor agent is selected from the list of anti-tumor agents in point (i) above.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與至少一種另外的抗腫瘤物質被同時地、分別地或順序地給予，該抗腫瘤物質選自順鉑、奧沙利鉑、卡鉑、戊柔比星、伊達比星、多柔比星、吡柔比星、伊立替康、托泊替康、胺柔比星、表柔比星、依託泊苷、絲裂黴素、苯達莫司汀、苯丁酸氮芥、環磷醯胺、異環磷醯胺、卡莫司汀、美法侖、博萊黴素、奧拉帕尼、MEDI4736、AZD1775以及AZD6738。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof Is administered simultaneously, separately or sequentially with at least one additional antitumor substance selected from the group consisting of cisplatin, oxaliplatin, carboplatin, penrorubicin, idarubicin, doxorubicin, Pirarubicin, irinotecan, topotecan, amrubicin, epirubicin, etoposide, mitomycin, bendamustine, chlorambucil, cyclophosphamide, Ifosfamide, carmustine, melphalan, bleomycin, olaparib, MEDI4736, AZD1775 and AZD6738.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用，其中具有化學式(I)之化合物或其藥學上可接受的鹽與至少一種另外的抗腫瘤物質被同時地、分別地或順序地給予，該抗腫瘤物質選自順鉑、奧沙利鉑、卡鉑、多柔比星、吡柔比星、伊立替康、托泊替康、胺柔比星、表柔比星、依託泊苷、絲裂黴素、苯達莫司汀、苯丁酸氮芥、環磷醯胺、異環磷醯胺、卡莫司汀、美法侖、博萊黴素、奧拉帕尼、AZD1775以及AZD6738。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is At least one additional antitumor substance is administered simultaneously, separately or sequentially, the antitumor substance is selected from the group consisting of cisplatin, oxaliplatin, carboplatin, doxorubicin, pirarubicin, irinotecan, and Porticon, Amrubicin, Epirubicin, Etoposide, Mitomycin, Bendamustine, Nitrogen mustard, Cyclophosphamide, Ifosfamide, Carmustine , Melphalan, bleomycin, olaparib, AZD1775 and AZD6738.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用，其中具有化學式(I)之化合物或其藥學上可接受的鹽與至少一種另外的抗腫瘤物質被同時地、分別地或順序地給予，該抗腫瘤物質選自多柔比星、伊立替康、托泊替康、依託泊苷、絲裂黴素、苯達莫司 汀、苯丁酸氮芥、環磷醯胺、異環磷醯胺、卡莫司汀、美法侖、博萊黴素以及奧拉帕尼。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is At least one additional anti-tumor substance is administered simultaneously, separately or sequentially, the anti-tumor substance is selected from doxorubicin, irinotecan, topotecan, etoposide, mitomycin, bendamer Stin, chlorambucil, cyclophosphamide, ifosfamide, carmustine, melphalan, bleomycin, and olaparib.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用，其中具有化學式(I)之化合物或其藥學上可接受的鹽與至少一種另外的抗腫瘤物質被同時地、分別地或順序地給予，該抗腫瘤物質選自多柔比星、伊立替康、托泊替康、依託泊苷、絲裂黴素、苯達莫司汀、苯丁酸氮芥、環磷醯胺、異環磷醯胺、卡莫司汀、美法侖以及博萊黴素。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is At least one additional anti-tumor substance is administered simultaneously, separately or sequentially, the anti-tumor substance is selected from doxorubicin, irinotecan, topotecan, etoposide, mitomycin, bendamer Stin, chlorambucil, cyclophosphamide, ifosfamide, carmustine, melphalan and bleomycin.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與至少一種另外的抗腫瘤物質被同時地、分別地或順序地給予，該抗腫瘤物質選自多柔比星、吡柔比星、胺柔比星以及表柔比星。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof It is administered simultaneously, separately or sequentially with at least one additional anti-tumor substance selected from doxorubicin, pirarubicin, amrubicin and epirubicin.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在急性髓性白血病的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與至少一種另外的抗腫瘤物質被同時地、分別地或順序地給予，該抗腫瘤物質選自多柔比星、吡柔比星、胺柔比星以及表柔比星。 In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of acute myeloid leukemia, wherein the compound of formula (I) or its pharmaceutically acceptable The received salt and at least one additional antitumor substance are administered simultaneously, separately or sequentially, the antitumor substance is selected from doxorubicin, pirarubicin, amrubicin and epirubicin.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在乳癌的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與至少一種另外的抗腫瘤物質被同時地、分別地或順序地給予，該抗腫瘤物質選自 多柔比星、吡柔比星、胺柔比星以及表柔比星。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of breast cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof It is administered simultaneously, separately or sequentially with at least one additional anti-tumor substance selected from doxorubicin, pirarubicin, amrubicin and epirubicin.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在三陰性乳癌的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與至少一種另外的抗腫瘤物質被同時地、分別地或順序地給予，該抗腫瘤物質選自多柔比星、吡柔比星、胺柔比星以及表柔比星。 In one embodiment, a salt with a compound of formula acceptable (I) or of a pharmaceutically acceptable for use in the treatment of triple negative breast cancer, wherein the compound having acceptable formula (I) or a pharmaceutically acceptable of The salt and at least one additional antitumor substance are administered simultaneously, separately or sequentially, the antitumor substance is selected from doxorubicin, pirarubicin, amrubicin and epirubicin.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在肝細胞癌的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與至少一種另外的抗腫瘤物質被同時地、分別地或順序地給予，該抗腫瘤物質選自多柔比星、吡柔比星、胺柔比星以及表柔比星。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of hepatocellular carcinoma, wherein the compound of formula (I) or a pharmaceutically acceptable The salt and at least one additional antitumor substance are administered simultaneously, separately or sequentially, the antitumor substance is selected from doxorubicin, pirarubicin, amrubicin and epirubicin.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與伊立替康被同時地、分別地或順序地給予。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof It is given simultaneously, separately or sequentially with irinotecan.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在結腸直腸癌的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與伊立替康被同時地、分別地或順序地給予。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of colorectal cancer, wherein the compound of formula (I) or a pharmaceutically acceptable The salt and irinotecan are administered simultaneously, separately or sequentially.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在結腸直腸癌的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與FOLFIRI 被同時地、分別地或順序地給予。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of colorectal cancer, wherein the compound of formula (I) or a pharmaceutically acceptable The salt and FOLFIRI are administered simultaneously, separately or sequentially.

FOLFIRI係包含亞葉酸、5-氟尿嘧啶以及伊立替康的組合的給藥方案。 FOLFIRI is a dosing regimen containing a combination of leucovorin, 5-fluorouracil, and irinotecan.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與奧拉帕尼被同時地、分別地或順序地給予。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof Olapani was given simultaneously, separately or sequentially.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在胃癌的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與奧拉帕尼被同時地、分別地或順序地給予。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of gastric cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof Olapani was given simultaneously, separately or sequentially.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與托泊替康被同時地、分別地或順序地給予。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof It is administered simultaneously, separately or sequentially with topotecan.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在肺癌的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與托泊替康被同時地、分別地或順序地給予。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of lung cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof It is administered simultaneously, separately or sequentially with topotecan.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在小細胞肺癌的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與托泊替康 被同時地、分別地或順序地給予。 In one embodiment, a salt with a compound of formula acceptable (I) or of a pharmaceutically acceptable for use in the treatment of small cell lung cancer, wherein the compound having acceptable formula (I) or a pharmaceutically acceptable of The salt and topotecan are administered simultaneously, separately or sequentially.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與免疫療法被同時地、分別地或順序地給予。在一個實施方式中，該免疫療法係列於以上點(iii)下的該等藥劑中的一種或多種。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof It is administered simultaneously, separately or sequentially with immunotherapy. In one embodiment, the immunotherapy series is one or more of the agents under point (iii) above.

在一個實施方式中，提供了具有化學式(I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用，其中該具有化學式(I)之化合物或其藥學上可接受的鹽與抗PD-L1抗體(例如MEDI4736)被同時地、分別地或順序地給予。 In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof The anti-PD-L1 antibody (eg MEDI4736) is administered simultaneously, separately or sequentially.

根據另一個實施方式，提供了套組，該套組包括：a)處於第一單位劑型的、具有化學式(I)之化合物或其藥學上可接受的鹽；b)處於另外的單位劑型的又另外的抗腫瘤物質；c)包含所述第一單位劑型和另外的單位劑型的容器裝置；以及視情況d)使用說明書。在一個實施方式中，該抗腫瘤物質包括抗腫瘤劑。 According to another embodiment, a kit is provided, the kit comprising: a) a compound of formula (I) or a pharmaceutically acceptable salt thereof in a first unit dosage form; b) in another unit dosage form Additional anti-tumor substance; c) a container device containing the first unit dosage form and another unit dosage form; and optionally d) instructions for use. In one embodiment, the anti-tumor substance includes an anti-tumor agent.

在抗腫瘤劑被提及的任何實施方式中，該抗腫瘤劑係列於以上點(i)下的該等藥劑中的一種或多種。 In any embodiment where the anti-neoplastic agent is mentioned, the anti-neoplastic agent series is one or more of the agents under point (i) above.

具有化學式(I)之化合物及其藥學上可接受的鹽 可以作為醫藥組成物被給予，該醫藥組成物包含一種或多種藥學上可接受的賦形劑。 The compound of formula (I) and its pharmaceutically acceptable salts can be administered as a pharmaceutical composition that includes one or more pharmaceutically acceptable excipients.

因此，在一個實施方式中，提供了包含具有化學式(I)之化合物或其藥學上可接受的鹽、以及至少一種藥學上可接受的賦形劑的醫藥組成物。 Therefore, in one embodiment, there is provided a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

針對包含於具體組成物中而選擇的一種或多種賦形劑將取決於如以下因素，如給予方式和提供的組成物的形式。適合的藥學上可接受的賦形劑係熟習該項技術者所熟知的並且例如，描述於《藥用賦形劑手冊》(Handbook of Pharmaceutical Excipients)中，第六版，英國醫藥出版社(Pharmaceutical Press)，由羅(Rowe)，雷(Ray)C；舍斯基(Sheskey)，保羅(Paul)J；奎恩(Quinn)，瑪麗安(Marian)編寫。藥學上可接受的賦形劑可以用作例如，佐劑、稀釋劑、載體、穩定劑、調味劑、著色劑、填料、粘合劑、崩散劑、潤滑劑、助流劑、增稠劑以及包衣劑。如熟習該項技術者將理解的是，某些藥學上可接受的賦形劑可用於多於一種功能，並且可用於可替代性作用，這取決於組成物中存在多少賦形劑並且該組成物中存在哪些其他賦形劑。 The excipient (s) selected for inclusion in a particular composition will depend on factors such as the mode of administration and the form of composition provided. Suitable pharmaceutically acceptable excipients are well known to those skilled in the art and are described, for example, in the Handbook of Pharmaceutical Excipients , Sixth Edition, British Medical Publishing House (Pharmaceutical Press), written by Rowe, Ray C; Sheskey, Paul J; Quinn, Marian. Pharmaceutically acceptable excipients can be used, for example, as adjuvants, diluents, carriers, stabilizers, flavoring agents, colorants, fillers, binders, disintegrating agents, lubricants, glidants, thickeners, Coating agent. As those skilled in the art will understand that certain pharmaceutically acceptable excipients can be used for more than one function and can be used for alternative effects, depending on how many excipients are present in the composition and the composition What other excipients are present in the substance.

該醫藥組成物可處於適合於以下的形式：口服使用(例如作為片劑、錠劑、硬或軟膠囊、水性或油性懸浮液、乳劑、可分散粉劑或顆粒劑、糖漿劑或酏劑)，局部使用(例如作為乳膏、軟膏劑、凝膠劑、或者水性或油性溶液或懸浮液)，藉由吸入給予(例如作為細碎粉末或液體氣霧劑)，藉由吹入給予(例如作為細碎粉末)，或腸胃外給予(例如作為用於靜脈內、皮下、肌內或肌 內給藥的無菌水性或油性溶液)，或作為用於直腸給藥給予的栓劑。該等組成物可以藉由本領域熟知的常規程序來獲得。旨在用於口服使用的組成物可含有另外的組分，例如，一種或多種著色劑、甜味劑、調味劑和/或防腐劑。 The pharmaceutical composition may be in a form suitable for oral use (for example as tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs), For topical use (eg as a cream, ointment, gel, or aqueous or oily solution or suspension), by inhalation (eg as a finely divided powder or liquid aerosol), by insufflation (eg as a finely divided Powder), or parenteral administration (for example, as a sterile aqueous or oily solution for intravenous, subcutaneous, intramuscular, or intramuscular administration), or as a suppository for rectal administration. These compositions can be obtained by conventional procedures well known in the art. Compositions intended for oral use may contain additional components, for example, one or more colorants, sweeteners, flavoring agents, and / or preservatives.

具有化學式(I)之化合物通常以範圍為2.5-5000mg/m2的動物體表面積內的單位劑量或約0.05-100mg/kg給予至溫血動物，並且這通常提供治療有效劑量。單位劑型如片劑或膠囊將通常含有例如0.1-250mg的活性成分。每日劑量將必然取決於所治療的宿主、具體的給予途徑、共給予的任何療法、以及正在治療的疾病的嚴重性而變化。因此，治療任何具體患者的執業醫生可以確定最佳劑量。 The compound of formula (I) is usually administered to a warm-blooded animal at a unit dose in the range of 2.5-5000 mg / m 2 of animal body surface area or about 0.05-100 mg / kg, and this generally provides a therapeutically effective dose. Unit dosage forms such as tablets or capsules will usually contain, for example, 0.1-250 mg of active ingredient. The daily dose will necessarily vary depending on the host being treated, the specific route of administration, any therapy co-administered, and the severity of the disease being treated. Therefore, a practicing doctor who treats any specific patient can determine the optimal dose.

本文所描述的該等醫藥組成物包含具有化學式(I)之化合物或其藥學上可接受的鹽，並且因此預期在療法中係有用的。 The pharmaceutical compositions described herein contain a compound of formula (I) or a pharmaceutically acceptable salt thereof, and are therefore expected to be useful in therapy.

同樣地，在一個實施方式中，提供了用於在療法中使用的醫藥組成物，該醫藥組成物包含具有化學式(I)之化合物或其藥學上可接受的鹽、以及至少一種藥學上可接受的賦形劑。 Similarly, in one embodiment, a pharmaceutical composition for use in therapy is provided, the pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable Of excipients.

在一個實施方式中，提供了用於在其中ATM激酶的抑制係有益的疾病的治療中使用的醫藥組成物，該醫藥組成物包含具有化學式(I)之化合物或其藥學上可接受的鹽、以及至少一種藥學上可接受的賦形劑。 In one embodiment, there is provided a pharmaceutical composition for use in the treatment of diseases in which inhibition of ATM kinase is beneficial, the pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, And at least one pharmaceutically acceptable excipient.

在一個實施方式中，提供了用於在癌症的治療中使 用的醫藥組成物，該醫藥組成物包含具有化學式(I)之化合物或其藥學上可接受的鹽、以及至少一種藥學上可接受的賦形劑。 In one embodiment, a pharmaceutical composition for use in the treatment of cancer is provided, the pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

在一個實施方式中，提供了用於在其中ATM激酶的抑制係有益的癌症的治療中使用的醫藥組成物，該醫藥組成物包含具有化學式(I)之化合物或其藥學上可接受的鹽、以及至少一種藥學上可接受的賦形劑。 In one embodiment, there is provided a pharmaceutical composition for use in the treatment of cancer in which inhibition of ATM kinase is beneficial, the pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, And at least one pharmaceutically acceptable excipient.

在一個實施方式中，提供了用於在治療以下疾病中使用的醫藥組成物：結腸直腸癌、惡性膠質瘤、胃癌、卵巢癌、彌漫性大B細胞淋巴瘤、慢性淋巴球性白血病、急性髓性白血病、頭頸部鱗狀細胞癌、乳癌、肝細胞癌、小細胞肺癌或非小細胞肺癌，該醫藥組成物包含具有化學式(I)之化合物或其藥學上可接受的鹽、以及至少一種藥學上可接受的賦形劑。 In one embodiment, a pharmaceutical composition for use in the treatment of the following diseases is provided: colorectal cancer, malignant glioma, gastric cancer, ovarian cancer, diffuse large B-cell lymphoma, chronic lymphocytic leukemia, acute myeloid Leukemia, squamous cell carcinoma of the head and neck, breast cancer, hepatocellular carcinoma, small cell lung cancer or non-small cell lung cancer, the pharmaceutical composition comprises a compound of formula (I) or a pharmaceutically acceptable salt thereof, and at least one pharmaceutical Acceptable excipient.

【實例】     【Example】    

藉由以下實例闡明本發明的多個實施方式。本發明不被解釋為受限於該等實例。在實例的製備期間，通常：i.除非另有說明，在環境溫度下即在約17℃至30℃的範圍內並且在惰性氣體的氣氛(如氮氣)下進行操作；ii.藉由旋轉蒸發或使用Genevac真空設備進行蒸發，並且在藉由過濾去除殘餘固體之後進行後處理常式；iii.在自動Armen Glider Flash：Spot II Ultimate(阿芒儀器(Armen Instrument)，聖阿韋(Saint-Ave)，法國)上或自動Presearch combiflash上伴隨使用從德國達姆施塔特的默克公司 (Merck,Darmstad,Germany)獲得的預包裝Merck正相Si60二氧化矽柱體(粒度計：15-40μm或40-63μm)、silicycle二氧化矽柱體或graceresolv二氧化矽柱體進行快速層析純化；iv.在裝有ZMD或ZQ ESCi質譜儀和沃特斯X-Terra反相柱或沃特斯X-Bridge反相柱或沃特斯SunFire反相柱(C-18，5微米二氧化矽，19mm或50mm直徑，100mm長度，40mL/分鐘的流速)的沃特斯儀器(600/2700或2525)上，使用水(含有1%氨)和乙腈的極性遞減混合物或者水(含有0.1%甲酸)和乙腈的極性遞減混合物作為洗提液進行製備型層析法；v.產率，在存在的情況下，不必是可達到的最大值；vi.具有化學式(I)之終產物的結構藉由核磁共振(NMR)譜證實，伴隨在δ規模上測量的NMR化學位移值。使用Bruker advance 700(700MHz)、Bruker Avance 500(500MHz)、Bruker 400(400Mhz)或Bruker 300(300Mhz)儀器測定質子核磁共振譜；在282Mhz或376Mhz處測定19F NMR；在75Mhz或100Mhz處測定13C NMR；除非另外指明，在大約20℃-30℃下進行測量；使用以下縮寫：s，單峰；d，二重峰；t，三重峰；q，四重峰；m，多重峰；dd，雙二重峰；ddd，雙二重峰的雙重峰；dt，雙三重峰；bs，寬信號；vii.具有化學式(I)之終產物還藉由液相層析質譜法(LCMS)來表徵；使用裝有沃特斯ZQ ESCi或ZMD ESCi質譜儀和X Bridge 5μm C-18柱(2.1 x 50mm)的沃特斯Alliance HT(2790 & 2795)在2.4mL/min的流速下，使用95% A+5% C至95% B+5% C的溶劑系統(其中A=水，B=甲醇，C=1：1甲醇：水(含有0.2%碳酸銨))經4分鐘；或藉由使用裝有Phenomenex Gemini-NX C18 3.0 x 50mm、3.0μM柱或等效物(鹼性條件)的Shimadzu UFLC或UHPLC外加DAD檢測器、ELSD檢測器和2020 Ev質譜儀(或等效物)，或Shim pack XR-ODS 3.0 x 50mm、2.2μM柱，或沃特斯BEH C18 2.1 x 50mm、1.7μM柱或等效物；使用95% D+5% E至95% E+5% D的溶劑系統(其中D=水(含有0.05% TFA)，E=乙腈(含有0.05% TFA)(酸性條件))經4分鐘或90% F+10% G至95% G+5% F的溶劑系統(其中F=水(含有6.5mM碳酸氫銨並且藉由添加氨調至pH 10)，G=乙腈(鹼性條件))經4分鐘進行LCMS；viii.中間物總體上未經完全表徵且純度藉由薄層層析、質譜、HPLC和/或NMR分析來評估；ix.藉由將結晶物質樣品安裝在Bruker單矽晶體(SSC)晶片支架上且借助於顯微鏡載片將樣品展布成薄層來測定(使用Bruker D4分析儀器)X射線粉末繞射光譜。使樣品以每分鐘30轉離心(以改良計數統計)且用由在40kV和40mA下操作的銅制長細聚焦管產生的具有1.5418埃的波長的X射線來照射。使准直X射線源穿過設定在V20下的自動可變發散狹縫且引導反射的輻射穿過5.89mm防散射狹縫和9.55mm檢測器狹縫。在θ-θ模式中從2°至40°2-θ的範圍內，使樣品每0.00570°2-θ增量(連續掃描模式)暴露0.03秒。執行時間係3分36秒。該儀器裝備有位置敏感性檢測器(聯凱)。對照和數據採集係藉由用Diffrac+軟體操作的Dell Optiplex 686 NT 4.0工作站進行的； x.在TA儀器Q1000 DSC上進行差示掃描熱量測定。典型地，將包含在裝配有蓋子的標準鋁盤中的小於5mg的材料以每分鐘10℃的恒定加熱速率在溫度範圍為25℃至300℃加熱。以每分鐘50ml流速使用用氮氣淨化的氣體；xi.使用以下縮寫：h=小時；r.t.=室溫(約18℃-25℃)；conc.=濃縮的；FCC=使用二氧化矽的快速柱層析法；DCM=二氯甲烷；DIPEA=二異丙基乙胺；DMA=N,N-二甲基乙醯胺；DMF=N,N-二甲基甲醯胺；DMSO=二甲基亞碸；Et2O=二***；EtOAc=乙酸乙酯；EtOH=乙醇；K2CO3=碳酸鉀；MeOH=甲醇；MeCN=乙腈；MTBE=甲基三級丁基醚；MgSO4=無水硫酸鎂；Na2SO4=無水硫酸鈉；THF=四氫呋喃；sat.=飽和水性溶液；並且xii.使用ELN(一種專利程式)、或者“Canvas”或“IBIS”(阿斯利康(AstraZeneca)專利程序)生成IUPAC名稱。如引言中所述的，本發明的該等化合物包括咪唑并[4,5-c]喹啉-2-酮核心。然而，在某些實施方式中，IUPAC名稱描述該核心為咪唑并[5,4-c]喹啉-2-酮或1H-咪唑并[4,5-c]喹啉-2(3H)-酮核心。儘管該咪唑并[4,5-c]喹啉-2-酮、1H-咪唑并[4,5-c]喹啉-2(3H)-酮和咪唑并[5,4-c]喹啉-2-酮核心係相同的，但是由於周邊基團，命名約定不同。 The following examples illustrate various embodiments of the present invention. The invention is not to be interpreted as being limited to these examples. During the preparation of the examples, usually: i. Unless otherwise stated, operate at ambient temperature, ie in the range of about 17 ° C to 30 ° C and under an inert gas atmosphere (such as nitrogen); ii. By rotary evaporation Or use Genevac vacuum equipment to evaporate and perform post-treatment routine after removing residual solids by filtration; iii. In automatic Armen Glider Flash: Spot II Ultimate (Armen Instrument (Armen Instrument), Saint-Ave ), France) or on an automatic Presearch combiflash with the use of pre-packed Merck normal phase Si60 silicon dioxide cylinders (particle size meter: 15-40μm) obtained from Merck, Darmstad, Germany Or 40-63μm), silicycle silica column or graceresolv silica column for flash chromatography purification; iv. When equipped with ZMD or ZQ ESCi mass spectrometer and Waters X-Terra reverse phase column or Waters Waters instrument (600/2700 or 2525) of X-Bridge reversed phase column or Waters SunFire reversed phase column (C-18, 5 micron silica, 19mm or 50mm diameter, 100mm length, 40mL / min flow rate) ), Use a decreasing polarity mixture of water (containing 1% ammonia) and acetonitrile Or a polar water (0.1% formic acid) and a mixture of acetonitrile decreasing Preparative chromatography as eluent;. V yields, in the presence of, not necessarily the maximum attainable;. Vi of formula (I ) The structure of the final product is confirmed by nuclear magnetic resonance (NMR) spectroscopy, accompanied by NMR chemical shift values measured on the delta scale. Proton nuclear magnetic resonance spectra were measured using Bruker advance 700 (700MHz), Bruker Avance 500 (500MHz), Bruker 400 (400Mhz), or Bruker 300 (300Mhz) instruments; 19F NMR was measured at 282Mhz or 376Mhz; ; Unless otherwise specified, measurement is performed at approximately 20 ° C-30 ° C; the following abbreviations are used: s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; dd, double Doublet; ddd, doublet of double doublet; dt, doublet triplet; bs, broad signal; vii. The final product of formula (I) is also characterized by liquid chromatography mass spectrometry (LCMS); Use Waters Alliance HT (2790 & 2795) equipped with Waters ZQ ESCi or ZMD ESCi mass spectrometer and X Bridge 5μm C-18 column (2.1 x 50mm) at a flow rate of 2.4mL / min, using 95% A + 5% C to 95% B + 5% C solvent system (where A = water, B = methanol, C = 1: 1 methanol: water (containing 0.2% ammonium carbonate)) for 4 minutes; or by using Shimadzu UFLC or UHPLC with Phenomenex Gemini-NX C18 3.0 x 50mm, 3.0μM column or equivalent (basic conditions) plus DAD detector, ELSD detector and 2020 Ev mass spectrometer (or equivalent), Shim pack XR-ODS 3.0 x 50mm, 2.2μM column, or Waters BEH C18 2.1 x 50mm, 1.7μM column or equivalent; use 95% D + 5% E to 95% E + 5% D solvent system (Where D = water (containing 0.05% TFA), E = acetonitrile (containing 0.05% TFA) (acidic conditions)) after 4 minutes or 90% F + 10% G to 95% G + 5% F solvent system (where F = water (containing 6.5 mM ammonium bicarbonate and adjusted to pH 10 by the addition of ammonia), G = acetonitrile (basic conditions) for 4 minutes; Liii; intermediates are generally not fully characterized and the purity is Evaluation by thin layer chromatography, mass spectrometry, HPLC, and / or NMR analysis; ix. By mounting a sample of crystalline material on a Bruker single silicon crystal (SSC) wafer holder and spreading the sample into a thin layer by means of a microscope slide The X-ray powder diffraction spectrum was measured (using Bruker D4 analytical instrument). The sample was centrifuged at 30 revolutions per minute (to improve count statistics) and irradiated with X-rays with a wavelength of 1.5418 Angstroms produced by a long thin copper focusing tube operated at 40 kV and 40 mA. The collimated X-ray source was passed through an automatic variable divergence slit set at V20 and the reflected radiation was guided through a 5.89 mm anti-scatter slit and a 9.55 mm detector slit. In the range of 2 ° to 40 ° 2-θ in the θ-θ mode, the sample is exposed for 0.03 seconds per 0.00570 ° 2-θ increment (continuous scanning mode). The execution time is 3 minutes and 36 seconds. The instrument is equipped with a position sensitive detector (Liankai). Control and data acquisition were performed by a Dell Optiplex 686 NT 4.0 workstation operated with Diffrac + software; x. Differential scanning calorimetry was performed on a TA instrument Q1000 DSC. Typically, less than 5 mg of material contained in a standard aluminum pan equipped with a lid is heated at a constant heating rate of 10 ° C per minute over a temperature range of 25 ° C to 300 ° C. Use a gas purged with nitrogen at a flow rate of 50 ml per minute; xi. Use the following abbreviations: h = hour; rt = room temperature (approximately 18 ° C to 25 ° C); conc. = Concentrated; FCC = flash column using silica Chromatography; DCM = dichloromethane; DIPEA = diisopropylethylamine; DMA = N , N -dimethylacetamide; DMF = N , N -dimethylformamide; DMSO = dimethyl Benzene chloride; Et 2 O = diethyl ether; EtOAc = ethyl acetate; EtOH = ethanol; K 2 CO 3 = potassium carbonate; MeOH = methanol; MeCN = acetonitrile; MTBE = methyl tertiary butyl ether; MgSO 4 = anhydrous Magnesium sulfate; Na 2 SO 4 = anhydrous sodium sulfate; THF = tetrahydrofuran; sat. = Saturated aqueous solution; and xii. Using ELN (a patented program), or “Canvas” or “IBIS” (AstraZeneca) patent (Procedure) Generate IUPAC name. As mentioned in the introduction, the compounds of the invention include imidazo [4,5-c] quinolin-2-one core. However, in certain embodiments, the IUPAC name describes the core as imidazo [5,4-c] quinolin-2-one or 1H-imidazo [4,5-c] quinolin-2 (3H)- Ketone core. Although the imidazo [4,5-c] quinolin-2-one, 1H-imidazo [4,5-c] quinolin-2 (3H) -one and imidazo [5,4-c] quinoline The 2-keto core system is the same, but due to the surrounding groups, the naming convention is different.

實例1 Example 1

(R)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮( R ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4 , 5-c] quinoline-2 (3H) -one

將二氯雙(二-三級丁基(3-磺丙基)磷鎓基)鈀酸鹽(II)(在水中的0.05M)(8.28mL，0.41mmol)添加到2-(甲氧基甲基)-5-(4,4,5,5-四甲基-1,3,2-二氧雜環戊硼烷(dioxaborolan)-2-基)吡啶(2.476g，9.94mmol)、(R)-8-溴-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮(3g，8.28mmol)以及2M K2CO3溶液(12.42mL，24.85mmol)在1,4-二(50mL)中的脫氣混合物中。將該混合物加熱至回流持續5小時。將該反應混合物用水稀釋，然後用EtOAc(3 x 100mL)萃取。將合併的有機相用水(100mL)、鹽水洗滌，並且然後經MgSO4乾燥，過濾並在真空中濃縮到二氧化矽上。將粗產物藉由FCC純化，洗提梯度為在DCM中的0至10%甲醇胺。將純級分蒸發至乾。將固體吸收於MeCN(25mL)中，在60℃下加熱1小時。藉由真空過濾收集固體，以得到呈白色固體的(R)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮(1.868g，55.8%)。NMR譜： 1H NMR(500MHz,DMSO-d6)δ 1.77-1.86(2H,m),2.16(1H,d),2.6-2.72(1H,m),3.34-3.4(1H,m),3.41(3H,s),3.49(3H,s),3.93(1H,d),4.13(1H,dd),4.21(1H,t),4.58(2H,s),4.93-5.04(1H,m),7.58(1H,d),7.99(1H,dd),8.17(1H,d),8.24(1H,dd),8.41(1H,d),8.90(1H,s),8.98(1H,d)。質譜：m/z：ES+[M+H]+ 405。 Add dichlorobis (di-tertiary butyl (3-sulfopropyl) phosphonium) palladium (II) (0.05M in water) (8.28mL, 0.41mmol) to 2- (methoxy) Methyl) -5- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) pyridine (2.476g, 9.94mmol), ( R ) -8-bromo-3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5-c] quinolin-2 (3H) -one (3g , 8.28mmol) and 2M K 2 CO 3 solution (12.42mL, 24.85mmol) in 1,4-two (50 mL) in the degassed mixture. The mixture was heated to reflux for 5 hours. The reaction mixture was diluted with water and then extracted with EtOAc (3 x 100 mL). The combined organic phase was washed with water (100 mL), brine, and then dried over MgSO 4 , filtered and concentrated onto silica in vacuo. The crude product was purified by FCC with an elution gradient of 0 to 10% methanolamine in DCM. The pure fractions were evaporated to dryness. The solid was absorbed in MeCN (25 mL) and heated at 60 ° C for 1 hour. The solid was collected by vacuum filtration to obtain ( R ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piper (Ran-3-yl) -1H-imidazo [4,5-c] quinolin-2 (3H) -one (1.868 g, 55.8%). NMR spectrum: 1 H NMR (500MHz, DMSO-d6) δ 1.77-1.86 (2H, m), 2.16 (1H, d), 2.6-2.72 (1H, m), 3.34-3.4 (1H, m), 3.41 ( 3H, s), 3.49 (3H, s), 3.93 (1H, d), 4.13 (1H, dd), 4.21 (1H, t), 4.58 (2H, s), 4.93-5.04 (1H, m), 7.58 (1H, d), 7.99 (1H, dd), 8.17 (1H, d), 8.24 (1H, dd), 8.41 (1H, d), 8.90 (1H, s), 8.98 (1H, d). Mass spectrum: m / z : ES + [M + H] + 405.

以類似方式從適合的硼酸酯和溴代中間體製備以下化合物。 The following compounds were prepared in a similar manner from suitable boronic esters and brominated intermediates.

*該反應使用二氯雙(二-三級丁基(3-磺丙基)磷鎓基)鈀酸鹽(II)(在水中的0.05M)作為催化劑，K2CO3作為鹼，並且在80℃下加熱2至4小時。 * This reaction uses dichlorobis (di-tertiarybutyl (3-sulfopropyl) phosphonium) palladium (II) (0.05M in water) as a catalyst, K 2 CO 3 as a base, and Heat at 80 ° C for 2 to 4 hours.

**該反應使用四(三苯基膦)鈀(0)作為催化劑，Cs2CO3作為鹼，並且在80℃下加熱18小時。 ** This reaction uses tetrakis (triphenylphosphine) palladium (0) as a catalyst, Cs 2 CO 3 as a base, and is heated at 80 ° C. for 18 hours.

***該反應使用四(三苯基膦)鈀(0)作為催化劑，Na2CO3作為鹼，並且在80℃下加熱16小時。 *** This reaction uses tetrakis (triphenylphosphine) palladium (0) as a catalyst, Na 2 CO 3 as a base, and is heated at 80 ° C. for 16 hours.

藉由製備型手性HPLC(Chiralpak AS柱)從外消旋混合物中分離實例4和5，用在EtOH(用三乙胺改性)中的80%庚烷作為洗提液進行等度洗提，以得到作為第一洗提產物的實例4和作為第二洗提產物的實例5。 Examples 4 and 5 were separated from the racemic mixture by preparative chiral HPLC (Chiralpak AS column) and isocratic elution was carried out using 80% heptane in EtOH (modified with triethylamine) as the eluent To obtain Example 4 as the first elution product and Example 5 as the second elution product.

藉由製備型HPLC(Chiralpak AD柱)分離實例6和7，用在EtOH-MeOH(用三乙胺改性)中的70%庚烷作為洗提液進行等度洗提。然後藉由製備型HPLC(Chiralpak AD柱)進一步純化該物質，用在EtOH-MeOH(用三乙胺改性)中的80%庚烷作為洗提液進行等度洗提，以得到作為第一洗提產物的實例6和作為第二洗提產物的實例7。 Examples 6 and 7 were separated by preparative HPLC (Chiralpak AD column) and isocratic elution was carried out using 70% heptane in EtOH-MeOH (modified with triethylamine) as the eluent. Then the material was further purified by preparative HPLC (Chiralpak AD column), isocratic elution was carried out using 80% heptane in EtOH-MeOH (modified with triethylamine) as the eluent to obtain as the first Example 6 of the eluted product and Example 7 as the second eluted product.

實例2 Example 2

NMR譜： 1H NMR(500MHz,DMSO-d6)δ 1.77-1.88(2H,m),2.16(1H,d),2.61-2.73(1H,m),3.35-3.4(1H,m),3.41(3H,s),3.48(3H,s),3.93(1H,d),4.1-4.17(1H,m),4.22(1H,t),4.58(2H,s),4.93-5.03(1H,m),7.58(1H,dd),7.98(1H,dd),8.16(1H,d), 8.24(1H,dd),8.41(1H,d),8.90(1H,s),8.98(1H,dd)。質譜：m/z：ES+[M+H]+ 405。 NMR spectrum: 1 H NMR (500MHz, DMSO-d6) δ 1.77-1.88 (2H, m), 2.16 (1H, d), 2.61-2.73 (1H, m), 3.35-3.4 (1H, m), 3.41 ( 3H, s), 3.48 (3H, s), 3.93 (1H, d), 4.1-4.17 (1H, m), 4.22 (1H, t), 4.58 (2H, s), 4.93-5.03 (1H, m) , 7.58 (1H, dd), 7.98 (1H, dd), 8.16 (1H, d), 8.24 (1H, dd), 8.41 (1H, d), 8.90 (1H, s), 8.98 (1H, dd). Mass spectrum: m / z : ES + [M + H] + 405.

(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮還可以藉由將在MeCN中的無定形固體(10mL/g)漿化而作為結晶固體來獲得。將漿液在60℃下加熱1小時，然後在室溫下攪拌過夜。藉由真空過濾收集結晶固體，以得到呈白色結晶固體的(S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮。使用X射線粉末繞射(XRPD)確認結晶度。實例2形式A用提供基本如圖1中所示出的X-射線粉末繞射圖而被表徵。十個X射線粉末繞射峰示於表1中： ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4 , 5-c] quinolin-2 (3H) -one can also be obtained as a crystalline solid by slurrying an amorphous solid (10 mL / g) in MeCN. The slurry was heated at 60 ° C for 1 hour and then stirred at room temperature overnight. The crystalline solid was collected by vacuum filtration to obtain ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H) as a white crystalline solid -Piperan-3-yl) -1H-imidazo [4,5-c] quinolin-2 (3H) -one. X-ray powder diffraction (XRPD) was used to confirm the crystallinity. Example 2 Form A was characterized by providing an X-ray powder diffraction pattern substantially as shown in FIG. Ten X-ray powder diffraction peaks are shown in Table 1:

實例2形式A展示了以下熱參數：如藉由差示掃描熱量測定(DSC)在10℃/min的掃描速率下所測定的以184.4℃開始的熔融吸熱和在186.0℃處的峰(圖2)。 Example 2 Form A demonstrates the following thermal parameters: as measured by differential scanning calorimetry (DSC) at a scan rate of 10 ° C / min, the endotherm of fusion starting at 184.4 ° C and the peak at 186.0 ° C (Figure 2 ).

實例3 Example 3

NMR譜： 1H NMR(500MHz,DMSO-d6)δ 1.66-1.87(2H,m),2.13(1H,d),2.57-2.73(1H,m),3.32-3.41(1H,m),3.42(3H,s),3.48(3H,s),3.82-3.97(1H,m),4.04-4.15(1H,m),4.19(1H,t),4.59(2H,s),4.79-5.06(1H,m),7.60(1H,dd),7.95(1H,d),8.16(1H,dt),8.30(1H,d),8.86(1H,s),8.93(1H,s)。質譜：m/z(ES+)[M+H]+=423。 NMR spectrum: 1 H NMR (500MHz, DMSO-d6) δ 1.66-1.87 (2H, m), 2.13 (1H, d), 2.57-2.73 (1H, m), 3.32-3.41 (1H, m), 3.42 ( 3H, s), 3.48 (3H, s), 3.82-3.97 (1H, m), 4.04-4.15 (1H, m), 4.19 (1H, t), 4.59 (2H, s), 4.79-5.06 (1H, m), 7.60 (1H, dd), 7.95 (1H, d), 8.16 (1H, dt), 8.30 (1H, d), 8.86 (1H, s), 8.93 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 423.

該材料還可以藉由溶解於少量水中並用溶解於少量水中的等量的甲磺酸處理，並且然後藉由冷凍乾燥去除水來分離為甲磺酸鹽。NMR譜： 1H NMR(300MHz,MeOH-d4)δ 1.83-1.98(2H,m),2.27(1H,d),2.71(3H,s),2.74-2.94(1H,m),3.46-3.54(1H,m),3.56(3H,s),3.61(3H,s),4.00(1H,dd),4.15-4.27(1H,m),4.38(1H,t),4.73(2H,s),5.02-5.19(1H,m),7.77-7.87(1H,m),8.02(1H,d),8.32-8.43(1H,m),8.59(1H,d),8.93(1H,d),9.09(1H,s)。質譜：m/z(ES+)[M+H]+=423。 The material can also be separated into mesylate by dissolving in a small amount of water and treating with an equal amount of methanesulfonic acid dissolved in a small amount of water, and then removing the water by freeze drying. NMR spectrum: 1 H NMR (300 MHz, MeOH-d4) δ 1.83-1.98 (2H, m), 2.27 (1H, d), 2.71 (3H, s), 2.74-2.94 (1H, m), 3.46-3.54 ( 1H, m), 3.56 (3H, s), 3.61 (3H, s), 4.00 (1H, dd), 4.15-4.27 (1H, m), 4.38 (1H, t), 4.73 (2H, s), 5.02 -5.19 (1H, m), 7.77-7.87 (1H, m), 8.02 (1H, d), 8.32-8.43 (1H, m), 8.59 (1H, d), 8.93 (1H, d), 9.09 (1H , s). Mass spectrum: m / z (ES +) [M + H] + = 423.

游離鹼(S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮也 可以作為結晶固體來獲得。藉由將該物質(20mg)在25℃下在四氫呋喃(2mL)中重結晶來生產實例3形式A。藉由將該物質(20mg)在環己烷(2mL)中漿化7天來生產實例3形式B。藉由在環境條件下蒸發來去除溶劑，以得到如藉由XRPD和DSC所測定的結晶物質。實例3形式A用提供基本如圖3中所示出的X-射線粉末繞射圖而被表徵。十個X射線粉末繞射峰示於表2中： Free base ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl)- 1,3-Dihydro-2H-imidazo [4,5-c] quinolin-2-one can also be obtained as a crystalline solid. Example 3 Form A was produced by recrystallizing this material (20 mg) in tetrahydrofuran (2 mL) at 25 ° C. Example 3 Form B was produced by slurrying this material (20 mg) in cyclohexane (2 mL) for 7 days. The solvent is removed by evaporation under ambient conditions to obtain crystalline material as determined by XRPD and DSC. Example 3 Form A was characterized by providing an X-ray powder diffraction pattern substantially as shown in FIG. Ten X-ray powder diffraction peaks are shown in Table 2:

實例3形式A展示了以下熱參數：如藉由差示掃描熱量測定(DSC)在10℃/min的掃描速率下所測定的以174.5℃開始的熔融吸熱和在177.3℃處的峰(圖4)。 Example 3 Form A shows the following thermal parameters: melting endotherm starting at 174.5 ° C and peak at 177.3 ° C as measured by differential scanning calorimetry (DSC) at a scan rate of 10 ° C / min (Figure 4 ).

實例3形式B用提供基本如圖5中所示出的X-射線粉末繞射圖 而被表徵。十個X射線粉末繞射峰示於表3中： Example 3 Form B was characterized by providing an X-ray powder diffraction pattern substantially as shown in FIG. Ten X-ray powder diffraction peaks are shown in Table 3:     

實例3形式B展示了以下熱參數：如藉由差示掃描熱量測定(DSC)在10℃/min的掃描速率下所測定的以174.5℃開始的熔融吸熱和在175.5℃處的峰(圖6)。 Example 3 Form B demonstrates the following thermal parameters: melting endotherm starting at 174.5 ° C and peak at 175.5 ° C as measured by differential scanning calorimetry (DSC) at a scan rate of 10 ° C / min (Figure 6 ).

實例4 Example 4

NMR譜： 1H NMR(400MHz,DMSO-d6)δ 1.71-1.82(2H,m),2.12(1H,d),2.56-2.68(1H,m),3.31-3.41(1H,m),3.45(3H,s),3.50(3H,s),3.88(1H,d),3.99-4.14(1H,m),4.24(1H,t),4.56(2H,s),4.79-4.95(1H,m),7.59(1H,dd),7.97(1H,d),8.29(1H,dd),8.37(1H,d),8.96(1H,s)。質譜：m/z(ES+)[M+H]+=441。 NMR spectrum: 1 H NMR (400 MHz, DMSO-d6) δ 1.71-1.82 (2H, m), 2.12 (1H, d), 2.56-2.68 (1H, m), 3.31-3.41 (1H, m), 3.45 ( 3H, s), 3.50 (3H, s), 3.88 (1H, d), 3.99-4.14 (1H, m), 4.24 (1H, t), 4.56 (2H, s), 4.79-4.95 (1H, m) , 7.59 (1H, dd), 7.97 (1H, d), 8.29 (1H, dd), 8.37 (1H, d), 8.96 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 441.

實例5 Example 5

NMR譜： 1H NMR(400MHz,DMSO-d6)δ 1.71-1.82(2H,m),2.12(1H,d),2.56-2.66(1H,m),3.32-3.42(1H,m),3.45(3H,s),3.50(3H,s),3.89(1H,d),3.99-4.14(1H,m),4.24(1H,t),4.56(2H,s),4.78-4.98(1H,m),7.58(1H,dd),7.97(1H,d),8.29(1H,dd),8.37(1H,d),8.96(1H,s)。質譜：m/z(ES+)[M+H]+=441 NMR spectrum: 1 H NMR (400 MHz, DMSO-d6) δ 1.71-1.82 (2H, m), 2.12 (1H, d), 2.56-2.66 (1H, m), 3.32-3.42 (1H, m), 3.45 ( 3H, s), 3.50 (3H, s), 3.89 (1H, d), 3.99-4.14 (1H, m), 4.24 (1H, t), 4.56 (2H, s), 4.78-4.98 (1H, m) , 7.58 (1H, dd), 7.97 (1H, d), 8.29 (1H, dd), 8.37 (1H, d), 8.96 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 441

實例6 Example 6

NMR譜： 1H NMR(400MHz,DMSO-d6)δ 1.78-1.86(2H,m),2.13-2.19(1H,m),2.62(1H,dd),3.35-3.43(1H,m),3.44(3H,s),3.51(3H,s),3.93(1H,d),4.05-4.17(1H,m),4.29(1H,t),4.54(2H,s),4.89-4.93(1H,m),7.56(1H,d),7.90(1H,d),8.18(1H,d),8.35(1H,dd),8.50(1H,s),8.94(1H,s)。質譜：m/z：ES+[M+H]+ 423 NMR spectrum: 1 H NMR (400MHz, DMSO-d6) δ 1.78-1.86 (2H, m), 2.13-2.19 (1H, m), 2.62 (1H, dd), 3.35-3.43 (1H, m), 3.44 ( 3H, s), 3.51 (3H, s), 3.93 (1H, d), 4.05-4.17 (1H, m), 4.29 (1H, t), 4.54 (2H, s), 4.89-4.93 (1H, m) , 7.56 (1H, d), 7.90 (1H, d), 8.18 (1H, d), 8.35 (1H, dd), 8.50 (1H, s), 8.94 (1H, s). Mass spectrum: m / z: ES + [M + H] + 423

實例7 Example 7

NMR譜： 1H NMR(400MHz,DMSO-d6)δ 1.83(2H,d),2.16(1H,d),2.58-2.64(1H,m),3.35-3.41(1H,m),3.44(3H,s),3.51(3H,s),3.93(1H,d),4.10(1H,d),4.29(1H,t),4.54(2H,s),4.84-5(1H,m),7.56(1H,d),7.90(1H,d),8.19(1H,d),8.35(1H,dd),8.50(1H,s),8.94(1H,s)。質譜：m/z：ES+[M+H]+ 423 NMR spectrum: 1 H NMR (400MHz, DMSO-d6) δ 1.83 (2H, d), 2.16 (1H, d), 2.58-2.64 (1H, m), 3.35-3.41 (1H, m), 3.44 (3H, s), 3.51 (3H, s), 3.93 (1H, d), 4.10 (1H, d), 4.29 (1H, t), 4.54 (2H, s), 4.84-5 (1H, m), 7.56 (1H , d), 7.90 (1H, d), 8.19 (1H, d), 8.35 (1H, dd), 8.50 (1H, s), 8.94 (1H, s). Mass spectrum: m / z: ES + [M + H] + 423

實例8 Example 8

NMR譜： 1H NMR(400MHz,MeOD-d4)δ 1.84-1.96(2H,m),2.24(1H,d),2.73-2.86(1H,m),3.50-3.58(1H,m),3.54(3H,s), 3.59(3H,s),4.00(1H,d),4.19(1H,d),4.39(1H,t),4.68(2H,s),4.96-5.09(1H,m),7.72(1H,d),7.91(1H,d),8.20-8.28(1H,m),8.44(1H,d),8.87(2H,s)。質譜：m/z(ES+),[M+H]+=423。 NMR spectrum: 1 H NMR (400 MHz, MeOD-d 4 ) δ 1.84-1.96 (2H, m), 2.24 (1H, d), 2.73-2.86 (1H, m), 3.50-3.58 (1H, m), 3.54 (3H, s), 3.59 (3H, s), 4.00 (1H, d), 4.19 (1H, d), 4.39 (1H, t), 4.68 (2H, s), 4.96-5.09 (1H, m), 7.72 (1H, d), 7.91 (1H, d), 8.20-8.28 (1H, m), 8.44 (1H, d), 8.87 (2H, s). Mass spectrum: m / z (ES +), [M + H] + = 423.

該材料還可以藉由溶解於少量水中並用溶解於少量水中的等量的甲磺酸處理，並且然後藉由冷凍乾燥去除水來分離為甲磺酸鹽。NMR譜： 1H NMR(300MHz,MeOD-d4)δ 1.83-1.97(2H,m),2.27(1H,d),2.70(3H,s),2.78-2.90(1H,m),3.49-3.59(1H,m),3.56(3H,s),3.61(3H,s),4.00(1H,d),4.15-4.27(1H,m),4.39(1H,t),4.73(2H,s),5.02-5.19(1H,m),7.82(1H,d),8.02(1H,d),8.31-8.42(1H,m),8.60(1H,d),8.93(1H,s),9.10(1H,s)。質譜：m/z(ES+),[M+H]+=423。 The material can also be separated into mesylate by dissolving in a small amount of water and treating with an equal amount of methanesulfonic acid dissolved in a small amount of water, and then removing the water by freeze drying. NMR spectrum: 1 H NMR (300 MHz, MeOD-d 4 ) δ 1.83-1.97 (2H, m), 2.27 (1H, d), 2.70 (3H, s), 2.78-2.90 (1H, m), 3.49-3.59 (1H, m), 3.56 (3H, s), 3.61 (3H, s), 4.00 (1H, d), 4.15-4.27 (1H, m), 4.39 (1H, t), 4.73 (2H, s), 5.02-5.19 (1H, m), 7.82 (1H, d), 8.02 (1H, d), 8.31-8.42 (1H, m), 8.60 (1H, d), 8.93 (1H, s), 9.10 (1H, s). Mass spectrum: m / z (ES +), [M + H] + = 423.

如下製備硼酸2-(甲氧基甲基)-5-(4,4,5,5-四甲基-1,3,2-二氧雜環戊硼烷-2-基)吡啶：2-(甲氧基甲基)-5-(4,4,5,5-四甲基-1,3,2-二氧雜環戊硼烷-2-基)吡啶 The 2- (methoxymethyl) -5- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) pyridine boric acid is prepared as follows: 2- (Methoxymethyl) -5- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) pyridine

將5-溴-2-(甲氧基甲基)吡啶(60g，297mmol)、4,4,4',4',5,5,5',5’-八甲基-2,2'-二(1,3,2-二氧雜環戊硼烷)(94g，371mmol)、乙酸鉀(87g，891mmol)和PdCl2(dppf)(0.217g，0.30mmol)在二(1L)中的攪拌混合物在氮氣下在100℃下攪拌16小時。將該混合物濃縮並通過矽藻土墊過濾，用己烷(1L)洗 提並蒸發，以給出呈棕色油狀物的2-(甲氧基甲基)-5-(4,4,5,5-四甲基-1,3,2-二氧雜環戊硼烷-2-基)吡啶(180g，162%)，將其不經進一步純化而使用。質譜：m/z：[M+H]+=250。 5-Bromo-2- (methoxymethyl) pyridine (60g, 297mmol), 4,4,4 ', 4', 5,5,5 ', 5'-octamethyl-2,2'- Di (1,3,2-dioxolane) (94g, 371mmol), potassium acetate (87g, 891mmol) and PdCl 2 (dppf) (0.217g, 0.30mmol) in The stirred mixture in (1L) was stirred under nitrogen at 100 ° C for 16 hours. The mixture was concentrated and filtered through a pad of diatomaceous earth, eluted with hexane (1 L) and evaporated to give 2- (methoxymethyl) -5- (4,4,5 as a brown oil , 5-tetramethyl-1,3,2-dioxaborolan-2-yl) pyridine (180 g, 162%), which was used without further purification. Mass spectrum: m / z: [M + H] + = 250.

5-溴-2-(甲氧基甲基)吡啶5-bromo-2- (methoxymethyl) pyridine

在0℃下，經10分鐘的時段，在氮氣下，將氫化鈉(4.98g，207mmol)分批添加至在THF(450mL)中的(5-溴吡啶-2-基)甲醇(30g，160mmol)中。將所得的溶液在25℃下攪拌0.5小時。在0℃下向該溶液中滴加碘甲烷(12.97mL，207mmol)。將所得的混合物在25℃下攪拌16小時。將該反應混合物用MeOH(20mL)稀釋。將粗產物藉由FCC純化，用在庚烷中的10%至60% EtOAc的梯度洗提。將純級分蒸發至乾燥，以得到呈黃色液體的5-溴-2-(甲氧基甲基)吡啶(30.0g，93%)。 1 H NMR譜(300MHz,CDCl3)：δ 3.48(3H,s),4.54(2H,s),7.34(1H,dd),7.83(1H,dd),8.61(1H,d)。 At 0 ° C, sodium hydride (4.98 g, 207 mmol) was added portionwise to (5-bromopyridin-2-yl) methanol (30 g, 160 mmol) in THF (450 mL) under nitrogen over a period of 10 minutes. )in. The resulting solution was stirred at 25 ° C for 0.5 hour. To this solution, iodomethane (12.97 mL, 207 mmol) was added dropwise at 0 ° C. The resulting mixture was stirred at 25 ° C for 16 hours. The reaction mixture was diluted with MeOH (20 mL). The crude product was purified by FCC, eluting with a gradient of 10% to 60% EtOAc in heptane. The pure fractions were evaporated to dryness to give 5-bromo-2- (methoxymethyl) pyridine (30.0 g, 93%) as a yellow liquid. 1 H NMR spectrum (300 MHz, CDCl 3 ): δ 3.48 (3H, s), 4.54 (2H, s), 7.34 (1H, dd), 7.83 (1H, dd), 8.61 (1H, d).

如下製備硼酸2-氟-6-(甲氧基甲基)-3-(4,4,5,5-四甲基-1,3,2-二氧雜環戊硼烷-2-基)吡啶：2-氟-6-(甲氧基甲基)-3-(4,4,5,5-四甲基-1,3,2-二氧雜環戊硼烷-2-基)吡啶 Boronate 2-fluoro-6- (methoxymethyl) -3- (4,4,5,5-tetramethyl-1,3,2-dioxolane-2-yl) was prepared as follows Pyridine: 2-fluoro-6- (methoxymethyl) -3- (4,4,5,5-tetramethyl-1,3,2-dioxaborol-2-yl) pyridine

將3-溴-2-氟-6-(甲氧基甲基)吡啶(2.2g，10.00mmol)、4,4,4',4',5,5,5',5’-八甲基-2,2'-二(1,3,2-二氧雜環戊硼烷)(2.79g，11.00mmol)、乙酸鉀(2.94g，30mmol)和{1,1'-雙(二苯基膦基)二茂鐵}氯化鈀與CH2Cl2(0.243g，0.30mmol)的複合物懸浮於脫氣的1,4-二(30mL)中。將該反應在氮氣氛下在80℃下加熱5小時。將粗混合物經矽藻土過濾，並且用MeCN、然後EtOAc洗滌。將濾液在真空中濃縮，以給出呈黑色固體的2-氟-6-(甲氧基甲基)-3-(4,4,5,5-四甲基-1,3,2-二氧雜環戊硼烷-2-基)吡啶(3.79g，142%)，將其不經進一步純化而使用。 1 H NMR譜(400MHz,DMSO-d6)：δ 1.30(12H,s),3.58(3H,s),4.45(2H,d),7.36(1H,dd),8.16(1H,t)。 3-Bromo-2-fluoro-6- (methoxymethyl) pyridine (2.2g, 10.00mmol), 4,4,4 ', 4', 5,5,5 ', 5'-octamethyl -2,2'-bis (1,3,2-dioxolane) (2.79g, 11.00mmol), potassium acetate (2.94g, 30mmol) and {1,1'-bis (diphenyl Phosphono) ferrocene} palladium chloride and CH 2 Cl 2 (0.243 g, 0.30 mmol) complex suspended in degassed 1,4-bis (30mL). The reaction was heated at 80 ° C for 5 hours under a nitrogen atmosphere. The crude mixture was filtered through celite and washed with MeCN, then EtOAc. The filtrate was concentrated in vacuo to give 2-fluoro-6- (methoxymethyl) -3- (4,4,5,5-tetramethyl-1,3,2-di as a black solid Oxaborolan-2-yl) pyridine (3.79 g, 142%), which was used without further purification. 1 H NMR spectrum (400 MHz, DMSO-d6): δ 1.30 (12H, s), 3.58 (3H, s), 4.45 (2H, d), 7.36 (1H, dd), 8.16 (1H, t).

3-溴-2-氟-6-(甲氧基甲基)吡啶3-bromo-2-fluoro-6- (methoxymethyl) pyridine

在-30℃下，在惰性氣氛下，經15分鐘將甲醇鈉(25%在MeOH中，2.472mL，10.81mmol)滴加到3-溴-6-(溴甲基)-2-氟吡啶(3.06g，11.38mmol)在乾DMF(20mL)中的攪拌溶液中。添加另外的甲醇鈉(0.15mL)，並攪拌該混合物直至反應完成。在-30℃下添加飽和水性碳酸氫鈉(約4mL)，並將該混合物在室溫下攪拌。將該混合物用DCM萃取，並且將有機相在真空中濃縮。將殘餘物藉由FCC進行純化，使用在己烷中的1%至1.5% EtOAc作為洗提液，以提供呈無色固體的3-溴-2-氟-6-(甲氧基甲基)吡啶(1.86g，74%)。 1 H NMR譜(400MHz,DMSO-d6)：3.37(3H,s), 4.43(2H,s),7.31(1H,d),8.30(1H,t)。 At -30 ° C, under an inert atmosphere, sodium methoxide (25% in MeOH, 2.472 mL, 10.81 mmol) was added dropwise to 3-bromo-6- (bromomethyl) -2-fluoropyridine ( 3.06 g, 11.38 mmol) in a stirred solution in dry DMF (20 mL). Additional sodium methoxide (0.15 mL) was added, and the mixture was stirred until the reaction was complete. Saturated aqueous sodium bicarbonate (about 4 mL) was added at -30 ° C, and the mixture was stirred at room temperature. The mixture was extracted with DCM, and the organic phase was concentrated in vacuo. The residue was purified by FCC using 1% to 1.5% EtOAc in hexane as eluent to provide 3-bromo-2-fluoro-6- (methoxymethyl) pyridine as a colorless solid (1.86g, 74%). 1 H NMR spectrum (400 MHz, DMSO-d6): 3.37 (3H, s), 4.43 (2H, s), 7.31 (1H, d), 8.30 (1H, t).

3-溴-6-(溴甲基)-2-氟吡啶3-bromo-6- (bromomethyl) -2-fluoropyridine

在黑暗中，將在四氯化碳(15mL)中的3-溴-2-氟-6-甲基吡啶(1.5g，7.89mmol)、N-溴代琥珀醯亞胺(1.62g，9.08mmol)和過氧化苯甲醯(0.191g，0.79mmol)在密封容器中在80℃下攪拌6小時。將該混合物冷卻至室溫並過濾。將濾液在真空中濃縮。藉由FCC純化，用在己烷中的2%至10% DCM進行洗提，以得到呈黃色油狀物的3-溴-6-(溴甲基)-2-氟吡啶(1.2g，56%)，其被22%起始材料污染。 1 H NMR譜(400MHz,DMSO-d6)4.68(2H,s),7.48(1H,dd),8.32(1H,dd)。質譜：m/z[M-H]-=268。 In the dark, combine 3-bromo-2-fluoro-6-picoline (1.5 g, 7.89 mmol) and N-bromosuccinimide (1.62 g, 9.08 mmol) in carbon tetrachloride (15 mL) ) And benzoyl peroxide (0.191 g, 0.79 mmol) were stirred in a sealed container at 80 ° C. for 6 hours. The mixture was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. Purified by FCC and eluted with 2% to 10% DCM in hexane to give 3-bromo-6- (bromomethyl) -2-fluoropyridine (1.2g, 56) as a yellow oil %), Which was contaminated with 22% starting material. 1 H NMR spectrum (400 MHz, DMSO-d6) 4.68 (2H, s), 7.48 (1H, dd), 8.32 (1H, dd). Mass spectrum: m / z [MH] - = 268.

如下製備溴中間體(S)-8-溴-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮：中間體A1：(S)-8-溴-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮 The bromine intermediate ( S ) -8-bromo-3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5-c] quinoline-2 ( 3H) -one: intermediate A1: ( S ) -8-bromo-3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5-c] quin Pyridin-2 (3H) -one

將碘甲烷(48.9g，344.63mmol)、四丁基溴化銨(2.78g，8.62mmol)和NaOH(10.34g，258.48mmol)在水(200mL)中的溶液添加到(S)-8-溴-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c] 喹啉-2(3H)-酮(30g，86.16mmol)在DCM(350mL)中的攪拌溶液中。將所得的溶液在25℃下攪拌16小時。將DCM在減壓下去除。藉由過濾收集沈澱物，用水(5 x 10mL)洗滌，並在真空烘箱中乾燥，以得到粗產物。將粗產物用EtOAc(150mL)稀釋。將所得的混合物在50℃下攪拌2小時。藉由過濾收集沈澱物，用EtOAc(5 x 5mL)洗滌並在真空烘箱中乾燥，以得到呈白色固體的(S)-8-溴-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮(25.00g，80%)。NMR譜：1H NMR(300MHz,DMSO-d6)δ 1.82-1.88(2H,m),2.09-2.15(1H,m),2.55-2.78(1H,m),3.30-3.47(1H,m)3.48(3H,s),3.92(1H,d),4.02-4.22(2H,m),4.68-4.88(1H,m),7.75(1H,d),7.99(1H,d),8.35(1H,s),8.92(1H,s)。質譜：m/z(ES+)[M+H]+=362。 A solution of methyl iodide (48.9g, 344.63mmol), tetrabutylammonium bromide (2.78g, 8.62mmol) and NaOH (10.34g, 258.48mmol) in water (200mL) was added to (S) -8-bromo -1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5-c] quinolin-2 (3H) -one (30g, 86.16mmol) in DCM (350mL) Stir the solution. The resulting solution was stirred at 25 ° C for 16 hours. The DCM was removed under reduced pressure. The precipitate was collected by filtration, washed with water (5 x 10 mL), and dried in a vacuum oven to obtain a crude product. The crude product was diluted with EtOAc (150 mL). The resulting mixture was stirred at 50 ° C for 2 hours. The precipitate was collected by filtration, washed with EtOAc (5 x 5 mL) and dried in a vacuum oven to give ( S ) -8-bromo-3-methyl-1- (tetrahydro-2H-piper (Ran-3-yl) -1H-imidazo [4,5-c] quinolin-2 (3H) -one (25.00 g, 80%). NMR spectrum: 1 H NMR (300 MHz, DMSO-d 6 ) δ 1.82-1.88 (2H, m), 2.09-2.15 (1H, m), 2.55-2.78 (1H, m), 3.30-3.47 (1H, m) 3.48 (3H, s), 3.92 (1H, d), 4.02-4.22 (2H, m), 4.68-4.88 (1H, m), 7.75 (1H, d), 7.99 (1H, d), 8.35 (1H, s), 8.92 (1H, s). Mass spectrum : m / z (ES +) [M + H] + = 362.

以類似方式從適當的3H-咪唑并[4,5-c]喹啉-2-酮中間體製備以下中間體： The following intermediates were prepared in a similar manner from the appropriate 3H-imidazo [4,5-c] quinolin-2-one intermediate:     

*反應尚未進行至完成，所以添加另外的碘甲烷、氫氧化鈉和四丁基溴化銨，並且將反應再攪拌16-18小時。 * The reaction has not proceeded to completion, so additional methyl iodide, sodium hydroxide and tetrabutylammonium bromide are added, and the reaction is stirred for an additional 16-18 hours.

**將反應在環境溫度下攪拌72小時。 ** The reaction was stirred at ambient temperature for 72 hours.

***使用2M NaOH溶液，並將反應在室溫下攪拌18小時。 *** 2M NaOH solution was used and the reaction was stirred at room temperature for 18 hours.

中間體B1： NMR譜：1H NMR(300MHz,DMSO-d6)δ 1.80-1.86(2H,m),2.07-2.12(1H,m),2.61-2.75(1H,m),3.32-3.46(1H,m),3.47(3H,s),3.92-3.98(1H,m),4.01-4.20(2H,m),4.72-4.83(1H,m),7.76(1H,dd),8.00(1H,d),8.34(1H,d),8.92(1H,s)。質譜：m/z(ES+)[M+H]+=362,364。 Intermediate B1: NMR spectrum : 1 H NMR (300 MHz, DMSO-d 6 ) δ 1.80-1.86 (2H, m), 2.07-2.12 (1H, m), 2.61-2.75 (1H, m), 3.32-3.46 ( 1H, m), 3.47 (3H, s), 3.92-3.98 (1H, m), 4.01-4.20 (2H, m), 4.72-4.83 (1H, m), 7.76 (1H, dd), 8.00 (1H, d), 8.34 (1H, d), 8.92 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 362,364.

中間體C1： NMR譜：1H NMR(400MHz,DMSO-d6)δ 1.88-190(2H,m),2.09(1H,d),2.70(1H,ddd),3.36-3.44(1H,m),3.47(3H,s),3.94(1H,d),4.07(1H,dd),4.15(1H,t),4.79(1H,ddd),7.97(1H,d),8.48(1H,d),8.93(1H,s)。質譜：m/z(ES+)[M+H]+=380,382。 Intermediate C1: NMR spectrum : 1 H NMR (400 MHz, DMSO-d 6 ) δ 1.88-190 (2H, m), 2.09 (1H, d), 2.70 (1H, ddd), 3.36-3.44 (1H, m) , 3.47 (3H, s), 3.94 (1H, d), 4.07 (1H, dd), 4.15 (1H, t), 4.79 (1H, ddd), 7.97 (1H, d), 8.48 (1H, d), 8.93 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 380,382.

中間體D1： NMR譜：1H NMR(400MHz,DMSO-d6)δ 1.86(2H, dd),2.11(1H,d),2.69(1H,ddd),3.37-3.45(1H,m),3.48(3H,s),3.95(1H,d),4.08(1H,dd),4.18(1H,t),4.80(1H,ddd),7.98(1H,d),8.50(1H,d),8.94(1H,s)。質譜：m/z(ES+)[M+H]+=380,382。 Intermediate D1: NMR spectrum : 1 H NMR (400 MHz, DMSO-d6) δ 1.86 (2H, dd), 2.11 (1H, d), 2.69 (1H, ddd), 3.37-3.45 (1H, m), 3.48 ( 3H, s), 3.95 (1H, d), 4.08 (1H, dd), 4.18 (1H, t), 4.80 (1H, ddd), 7.98 (1H, d), 8.50 (1H, d), 8.94 (1H , s). Mass spectrum: m / z (ES +) [M + H] + = 380,382.

中間體E1： NMR譜：1H NMR(400MHz,DMSO-d6)δ 1.77-1.92(2H,m),2.11(1H,d),2.62-2.77(1H,m),3.40(1H,td),3.49(3H,s),3.95(1H,d),4.08(1H,dd),4.18(1H,t),4.68-4.9(1H,m),7.77(1H,dd),8.01(1H,d),8.36(1H,d),8.93(1H,s)。質譜：m/z(ES+)[M+H]+=362,364。 Intermediate E1: NMR spectrum : 1 H NMR (400 MHz, DMSO-d 6 ) δ 1.77-1.92 (2H, m), 2.11 (1H, d), 2.62-2.77 (1H, m), 3.40 (1H, td) , 3.49 (3H, s), 3.95 (1H, d), 4.08 (1H, dd), 4.18 (1H, t), 4.68-4.9 (1H, m), 7.77 (1H, dd), 8.01 (1H, d ), 8.36 (1H, d), 8.93 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 362,364.

中間體F1： NMR譜：1H NMR(400MHz,DMSO-d6)δ 1.8-1.92(2H,m),2.11(1H,d),2.64-2.75(1H,m),3.39-3.43(1H,m),3.48(3H,s),3.95(1H,d),4-4.12(1H,m),4.18(1H,t),4.78-4.86(1H,m),7.99(1H,d),8.51(1H,d),8.95(1H,s)。質譜：m/z(ES+)[M+H]+=380,382。 Intermediate F1: NMR spectrum: 1 H NMR (400 MHz, DMSO-d6) δ 1.8-1.92 (2H, m), 2.11 (1H, d), 2.64-2.75 (1H, m), 3.39-3.43 (1H, m ), 3.48 (3H, s), 3.95 (1H, d), 4-4.12 (1H, m), 4.18 (1H, t), 4.78-4.86 (1H, m), 7.99 (1H, d), 8.51 ( 1H, d), 8.95 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 380,382.

如下製備3H-咪唑并[4,5-c]喹啉-2-酮中間體(S)-8-溴-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮：中間體A2：(S)-8-溴-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮 The 3H-imidazo [4,5-c] quinolin-2-one intermediate ( S ) -8-bromo-1- (tetrahydro-2H-piperan-3-yl) -1,3-di was prepared as follows Hydrogen-2H-imidazo [4,5-c] quinolin-2-one: intermediate A2: ( S ) -8-bromo-1- (tetrahydro-2H-piperan-3-yl) -1, 3-dihydro-2H-imidazo [4,5-c] quinolin-2-one

將三乙胺(17.86mL，128.13mmol)添加至在DMF(100mL)中的(S)-6-溴-4-((四氫-2H-哌喃-3-基)胺基)喹啉-3-甲酸(15g，42.71mmol)中。將所得的懸浮液攪拌25分鐘，然後冷卻 至0℃。滴加二苯基磷醯基疊氮化物(11.05mL，51.25mmol)，保持溫度低於10℃。將該懸浮液在室溫下再攪拌30分鐘，然後在60℃下攪拌2小時。將該反應混合物冷卻至室溫，添加水(100mL)並攪拌30分鐘。藉由過濾收集所得的沈澱物，以得到呈灰白色固體的(S)-8-溴-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮(13.20g，89%)。NMR譜：1H NMR(300MHz,DMSO-d6)δ 1.84-2.11(3H,m),2.62-2.76(1H,m),3.35-3.44(1H,m),3.92-4.22(3H,m),4.71-4.80(1H,m),7.76(1H,dd),7.98(2H,d),8.32(1H,dd),8.71(1H,s),11.85(1H,bs)。質譜：m/z(ES+)[M+H]+=348,350。 Triethylamine (17.86 mL, 128.13 mmol) was added to ( S ) -6-bromo-4-((tetrahydro-2H-piperan-3-yl) amino) quinoline in DMF (100 mL) 3-carboxylic acid (15g, 42.71mmol). The resulting suspension was stirred for 25 minutes and then cooled to 0 ° C. Add diphenylphosphoryl azide (11.05 mL, 51.25 mmol) dropwise, keeping the temperature below 10 ° C. The suspension was stirred at room temperature for another 30 minutes and then at 60 ° C for 2 hours. The reaction mixture was cooled to room temperature, water (100 mL) was added and stirred for 30 minutes. The resulting precipitate was collected by filtration to obtain ( S ) -8-bromo-1- (tetrahydro-2H-piperan-3-yl) -1,3-dihydro-2H-imidazo as an off-white solid [4,5-c] quinolin-2-one (13.20 g, 89%). NMR spectrum : 1 H NMR (300 MHz, DMSO-d6) δ 1.84-2.11 (3H, m), 2.62-2.76 (1H, m), 3.35-3.44 (1H, m), 3.92-4.22 (3H, m), 4.71-4.80 (1H, m), 7.76 (1H, dd), 7.98 (2H, d), 8.32 (1H, dd), 8.71 (1H, s), 11.85 (1H, bs). Mass spectrum: m / z (ES +) [M + H] + = 348,350.

以類似方式從適當的羧酸中間體製備以下3H-咪唑并[4,5-c]喹啉-2-酮中間體： **將反應在60℃下攪拌60分鐘-90分鐘。 The following 3H-imidazo [4,5-c] quinolin-2-one intermediates were prepared in a similar manner from the appropriate carboxylic acid intermediates:      ** The reaction was stirred at 60 ° C for 60 minutes to 90 minutes.

中間體B2： NMR譜：1H NMR(300MHz,DMSO-d6)δ 1.82-2.11(3H,m),2.61-2.75(1H,m),3.34-3.43(1H,m),3.91-4.21(3H,m),4.69-4.78(1H,m),7.75(1H,dd),7.99(2H,d),8.33(1H,dd),8.69(1H,s),11.70(1H,bs)。質譜：m/z(ES+)[M+H]+=348,350。 Intermediate B2: NMR spectrum: 1 H NMR (300 MHz, DMSO-d 6 ) δ 1.82-2.11 (3H, m), 2.61-2.75 (1H, m), 3.34-3.43 (1H, m), 3.91-4.21 ( 3H, m), 4.69-4.78 (1H, m), 7.75 (1H, dd), 7.99 (2H, d), 8.33 (1H, dd), 8.69 (1H, s), 11.70 (1H, bs). Mass spectrum: m / z (ES +) [M + H] + = 348,350.

中間體C2： NMR譜：1H NMR(400MHz,DMSO-d6)δ 1.77-1.93(2H,m),2.10(1H,d),2.68(1H,qd),3.34-3.44(1H,m),3.94(1H,d),4.08(1H,dd),4.18(1H,t),4.75(1H,ddd),7.94(1H,d),8.48(1H,d),8.69(1H,s),11.63(1H,s)。質譜：m/z(ES+)[M+H]+=366,368。 Intermediate C2: NMR spectrum : 1 H NMR (400 MHz, DMSO-d 6 ) δ 1.77-1.93 (2H, m), 2.10 (1H, d), 2.68 (1H, qd), 3.34-3.44 (1H, m) , 3.94 (1H, d), 4.08 (1H, dd), 4.18 (1H, t), 4.75 (1H, ddd), 7.94 (1H, d), 8.48 (1H, d), 8.69 (1H, s), 11.63 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 366,368.

中間體D2： NMR譜：1H NMR(400MHz,DMSO-d6)δ 1.7-1.93(2H,m),2.10(1H,d),2.63-2.75(1H,m),3.49-3.61(1H,m),3.84-4.03(1H,m),4.08(1H,dd),4.19(1H,t),4.76(1H,t),7.95(1H,d),8.49(1H,d),8.70(1H,s),11.66(1H,s)。質譜：m/z(ES+)[M+H]+=366,368。 Intermediate D2: NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ) δ 1.7-1.93 (2H, m), 2.10 (1H, d), 2.63-2.75 (1H, m), 3.49-3.61 (1H, m), 3.84-4.03 (1H, m), 4.08 (1H, dd), 4.19 (1H, t), 4.76 (1H, t), 7.95 (1H, d), 8.49 (1H, d), 8.70 (1H , s), 11.66 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 366,368.

中間體E2：質譜：m/z(ES+)[M+H]+=348。 Intermediate E2: Mass spectrum: m / z (ES +) [M + H] + = 348.

如下製備羧酸中間體(S)-6-溴-4-((四氫-2H-哌喃-3-基)胺基)喹啉-3-甲酸：中間體A3：(S)-6-溴-4-((四氫-2H-哌喃-3-基)胺基)喹啉-3-甲酸 The carboxylic acid intermediate ( S ) -6-bromo-4-((tetrahydro-2H-piperan-3-yl) amino) quinoline-3-carboxylic acid was prepared as follows: Intermediate A3: ( S ) -6- Bromo-4-((tetrahydro-2H-piperan-3-yl) amino) quinoline-3-carboxylic acid

將(S)-6-溴-4-((四氫-2H-哌喃-3-基)胺基)喹啉-3-甲酸乙酯(27.8g，73.30mmol)在THF(400mL)和水(80mL)中的溶液用2M氫氧化鈉(73.3mL，146.60mmol)處理，並在60℃下攪拌6小時，然後允許冷卻至室溫。在減壓下去除溶劑，並且將水溶液用2M HCl調節至pH 3，給出沈澱物，將該沈澱物濾出，用水充分洗滌並乾燥，以得到呈白色固體的(S)-6-溴-4-((四氫-2H-哌喃-3-基)胺基)喹啉-3-甲酸(19.05g，74.0%)。NMR譜：1H NMR(300MHz,DMSO-d6)δ 1.50-1.57(1H,m),1.61-1.82(2H,m),1.98-2.13(1H,m),3.48-3.72(3H,m),3.89(1H,d),4.15-4.26(1H,m),7.77(1H,dd),7.95(1H,d),8.31(1H,d),8.90(1H,s),13.38(1H,bs)。質譜：m/z(ES+)[M+H]+=351。 ( S ) -6-Bromo-4-((tetrahydro-2H-piperan-3-yl) amino) quinoline-3-carboxylic acid ethyl ester (27.8g, 73.30mmol) in THF (400mL) and water The solution in (80 mL) was treated with 2M sodium hydroxide (73.3 mL, 146.60 mmol) and stirred at 60 ° C for 6 hours, then allowed to cool to room temperature. The solvent was removed under reduced pressure, and the aqueous solution was adjusted to pH 3 with 2M HCl to give a precipitate, which was filtered off, washed thoroughly with water and dried to give ( S ) -6-bromo- as a white solid 4-((Tetrahydro-2H-piperan-3-yl) amino) quinoline-3-carboxylic acid (19.05 g, 74.0%). NMR spectrum: 1 H NMR (300 MHz, DMSO-d 6 ) δ 1.50-1.57 (1H, m), 1.61-1.82 (2H, m), 1.98-2.13 (1H, m), 3.48-3.72 (3H, m) , 3.89 (1H, d), 4.15-4.26 (1H, m), 7.77 (1H, dd), 7.95 (1H, d), 8.31 (1H, d), 8.90 (1H, s), 13.38 (1H, bs ). Mass spectrum : m / z (ES +) [M + H] + = 351.

以類似方式從適當的酯先質製備以下羧酸中間體： The following carboxylic acid intermediates are prepared in a similar manner from the appropriate ester precursor:     

*將反應在60℃-70℃之間攪拌1小時-3小時。 * Stir the reaction at 60 ° C-70 ° C for 1 hour to 3 hours.

***使用THF和水的混合物作為溶劑進行反應，並且在60℃下加熱3小時-16小時。 *** The reaction is performed using a mixture of THF and water as a solvent, and heated at 60 ° C for 3-16 hours.

中間體B3： NMR譜：1H NMR(300MHz,DMSO-d6)δ 1.50-1.56(1H,m),1.62-1.83(2H,m),1.99-2.12(1H,m),3.50-3.71(3H,m),3.89(1H,d),4.16-4.28(1H,m),7.78(1H,dd),7.94(1H,d),8.30(1H,d),8.94(1H,s),13.50(1H,bs)。質譜：m/z(ES+)[M+H]+=351。 Intermediate B3: NMR spectrum: 1 H NMR (300 MHz, DMSO-d 6 ) δ 1.50-1.56 (1H, m), 1.62-1.83 (2H, m), 1.99-2.12 (1H, m), 3.50-3.71 ( 3H, m), 3.89 (1H, d), 4.16-4.28 (1H, m), 7.78 (1H, dd), 7.94 (1H, d), 8.30 (1H, d), 8.94 (1H, s), 13.50 (1H, bs). Mass spectrum : m / z (ES +) [M + H] + = 351.

中間體C3： NMR譜：1H NMR(300MHz,DMSO-d6)δ 1.51(1H,m),1.74(2H,m),2.04(1H,m),3.60(3H,m),3.82(1H,d),4.15(1H,m),7.73(1H,m),8.44(1H,m),8.92(1H,s)。質譜：m/z(ES+)[M+H]+=369。 Intermediate C3: NMR spectrum: 1 H NMR (300 MHz, DMSO-d 6 ) δ 1.51 (1H, m), 1.74 (2H, m), 2.04 (1H, m), 3.60 (3H, m), 3.82 (1H , d), 4.15 (1H, m), 7.73 (1H, m), 8.44 (1H, m), 8.92 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 369.

中間體D3： 質譜：m/z(ES+)[M+H]+=369。 Intermediate D3: Mass spectrum: m / z (ES +) [M + H] + = 369.

中間體E3： 質譜：m/z(ES+)[M+H]+=351。 Intermediate E3: Mass spectrum: m / z (ES +) [M + H] + = 351.

如下製備酯中間體(S)-6-溴-4-((四氫-2H-哌喃-3-基)胺基)喹啉-3-甲酸乙酯：中間體A4：(S)-6-溴-4-((四氫-2H-哌喃-3-基)胺基)喹啉-3-甲酸乙酯 The ester intermediate ( S ) -6-bromo-4-((tetrahydro-2H-piperan-3-yl) amino) quinoline-3-carboxylic acid ethyl ester was prepared as follows: Intermediate A4: ( S ) -6 -Bromo-4-((tetrahydro-2H-piperan-3-yl) amino) quinoline-3-carboxylic acid ethyl ester

在25℃下，在空氣下，將DIPEA(6.99mL，40.00 mmol)添加到在DMA(30mL)中的6-溴-4-氯喹啉-3-甲酸乙酯(3.15g，10mmol)和(S)-四氫-2H-哌喃-3-胺鹽酸鹽(1.376g，10.00mmol)中。將所得的溶液在80℃下攪拌16小時。將該反應混合物用水(100mL)稀釋，藉由過濾收集沈澱物，用水(20mL)洗滌並溶解於250mL EtOAc/DCM(1：1)中。將形成的混合物經MgSO4乾燥，過濾並蒸發，以得到粗的呈白色固體的(S)-6-溴-4-((四氫-2H-哌喃-3-基)胺基)喹啉-3-甲酸乙酯(3.16g，83%)。將該產物不經進一步純化直接用於下一步驟。NMR譜：1H NMR(300MHz,DMSO-d6)δ 1.36(3H,t),1.70-1.74(1H,m),1.75-1.77(2H,m),2.03-2.05(1H,m),3.58-3.61(3H,m),3.80-3.85(1H,m),4.01-4.03(1H,m),4.35(2H,q),7.80(1H,d),7.89(1H,dd),8.58(1H,s),8.67(1H,d),8.93(1H,s)。質譜：m/z：ES+[M+H]+ 379,381。 At 25 ° C, under air, DIPEA (6.99 mL, 40.00 mmol) was added to ethyl 6-bromo-4-chloroquinoline-3-carboxylate (3.15 g, 10 mmol) and ( S ) -Tetrahydro-2H-piperan-3-amine hydrochloride (1.376 g, 10.00 mmol). The resulting solution was stirred at 80 ° C for 16 hours. The reaction mixture was diluted with water (100 mL), the precipitate was collected by filtration, washed with water (20 mL) and dissolved in 250 mL EtOAc / DCM (1: 1). The resulting mixture was dried over MgSO 4 , filtered and evaporated to give crude ( S ) -6-bromo-4-((tetrahydro-2H-piperan-3-yl) amino) quinoline as a white solid Ethyl-3-carboxylate (3.16g, 83%). This product was used directly in the next step without further purification. NMR spectrum : 1 H NMR (300 MHz, DMSO-d 6 ) δ 1.36 (3H, t), 1.70-1.74 (1H, m), 1.75-1.77 (2H, m), 2.03-2.05 (1H, m), 3.58 -3.61 (3H, m), 3.80-3.85 (1H, m), 4.01-4.03 (1H, m), 4.35 (2H, q), 7.80 (1H, d), 7.89 (1H, dd), 8.58 (1H , s), 8.67 (1H, d), 8.93 (1H, s). Mass spectrum: m / z : ES + [M + H] + 379,381.

以類似的方式從適當的胺以及6-溴-4-氯-7-氟喹啉-3-甲酸乙酯或6-溴-4-氯喹啉-3-甲酸乙酯製備以下酯中間體： The following ester intermediates were prepared in a similar manner from the appropriate amine and ethyl 6-bromo-4-chloro-7-fluoroquinoline-3-carboxylate or ethyl 6-bromo-4-chloroquinoline-3-carboxylate:     

*將反應在80℃下攪拌2小時-16小時 * Stir the reaction at 80 ° C for 2-16 hours

**將反應在100℃下攪拌16小時 ** Stir the reaction at 100 ° C for 16 hours

中間體B4：質譜：m/z(ES+)[M+H]+=379,381。 Intermediate B4: Mass spectrum: m / z (ES +) [M + H] + = 379,381.

中間體C4： NMR譜：1H NMR(300MHz,DMSO-d6)δ 1.33(3H,m),1.51(1H,m),1.74(2H,m),2.04(1H,m),3.60(3H,m),3.82(1H,d),4.02(1H,m),4.35(2H,m),7.73(1H,m),8.49(1H,m),8.79(1H,m),8.88(1H,s)。質譜：m/z(ES+)[M+H]+=397。 Intermediate C4: NMR spectrum: 1 H NMR (300 MHz, DMSO-d6) δ 1.33 (3H, m), 1.51 (1H, m), 1.74 (2H, m), 2.04 (1H, m), 3.60 (3H, m), 3.82 (1H, d), 4.02 (1H, m), 4.35 (2H, m), 7.73 (1H, m), 8.49 (1H, m), 8.79 (1H, m), 8.88 (1H, s ). Mass spectrum: m / z (ES +) [M + H] + = 397.

中間體D4： NMR譜：1H NMR(400MHz,CDCl3)δ 1.44(3H,t),1.67-1.77(1H,m),1.86-1.97(2H,m),2.17-2.24(1H,m),3.60-3.65(1H,m),3.72-3.78(2H,m),3.93-4.02(1H,m),4.12-4.19(1H,m),4.44(2H,q)7.72(1H,d),8.32(1H,d),9.14(1H,s),9.46(1H,s)。質譜：m/z(ES+)[M+H]+=397。 Intermediate D4: NMR spectrum : 1 H NMR (400 MHz, CDCl 3 ) δ 1.44 (3H, t), 1.67-1.77 (1H, m), 1.86-1.97 (2H, m), 2.17-2.24 (1H, m) , 3.60-3.65 (1H, m), 3.72-3.78 (2H, m), 3.93-4.02 (1H, m), 4.12-4.19 (1H, m), 4.44 (2H, q) 7.72 (1H, d), 8.32 (1H, d), 9.14 (1H, s), 9.46 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 397.

中間體E4：質譜：m/z(ES+)[M+H]+=379。 Intermediate E4: Mass spectrum: m / z (ES +) [M + H] + = 379.

中間體A5：6-溴-4-氯喹啉-3-甲酸乙酯Intermediate A5: ethyl 6-bromo-4-chloroquinoline-3-carboxylate

在環境溫度下，在空氣下，將DMF(0.119mL，1.54mmol)添加至在亞硫醯氯(800mL)中的6-溴-1-[(4-甲氧基苯基) 甲基]-4-側氧基喹啉-3-甲酸乙酯(160g，384.37mmol)中。將所得的混合物在75℃下攪拌16小時，然後將溶劑在減壓下去除。將所得的混合物與甲苯共沸兩次，然後添加正己烷(500mL)。藉由真空過濾收集沈澱物，用正己烷(200mL)洗滌並在真空下乾燥，以得到呈棕色固體的6-溴-4-氯喹啉-3-甲酸乙酯(100g，83%)。NMR譜：1H NMR(400MHz,CDCl3)δ 1.47(3H,t),4.51(2H,q),7.95(1H,dd),8.11(1H,d),8.60(1H,d),9.24(1H,s)。質譜：m/z(ES+)[M+H]+=314,316。 At ambient temperature, under air, DMF (0.119 mL, 1.54 mmol) was added to 6-bromo-1-[(4-methoxyphenyl) methyl]-in thiosulfonyl chloride (800 mL)- 4-oxoquinoline-3-carboxylic acid ethyl ester (160 g, 384.37 mmol). The resulting mixture was stirred at 75 ° C for 16 hours, and then the solvent was removed under reduced pressure. The resulting mixture was azeotroped twice with toluene, and then n-hexane (500 mL) was added. The precipitate was collected by vacuum filtration, washed with n-hexane (200 mL) and dried under vacuum to obtain ethyl 6-bromo-4-chloroquinoline-3-carboxylate (100 g, 83%) as a brown solid. NMR spectrum: 1 H NMR (400 MHz, CDCl 3 ) δ 1.47 (3H, t), 4.51 (2H, q), 7.95 (1H, dd), 8.11 (1H, d), 8.60 (1H, d), 9.24 ( 1H, s). Mass spectrum: m / z (ES +) [M + H] + = 314,316.

以更大的規模，將6-溴-1-[(4-甲氧基苯基)甲基]-4-側氧基喹啉-3-甲酸乙酯(5765g，13.85mol)填裝至具有亞硫醯氯(28.8L)的容器中。觀察到從20℃-26℃的放熱。添加DMF(4.4mL)，沒有觀察到放熱，並且將該批次加熱至75℃並攪拌17小時。HPLC顯示98.0%產物有1.3%起始物質剩餘。將該反應在真空中濃縮並將殘餘物與甲苯(25L)共沸。然後將所得的固體在庚烷(18.5L)中漿化2.5小時，過濾並用庚烷(3 x 4L)洗滌。將固體在真空下在35℃下乾燥以給出4077g所希望的物質(93%粗產量)，藉由HPLC，該物質除了約4%水解產物之外，還含有約5%的6-溴-1-[(4-甲氧基苯基)甲基]-4-側氧基喹啉-3-甲酸乙酯(90%純)。將該粗物質(4077g)返回至容器中並用亞硫醯氯(14.5L)和DMF(2.2mL)重新處理。將該混合物加熱至75℃持續40小時。將亞硫醯氯在真空中去除並且將殘餘物與甲苯(10L)共沸。在20℃下，將該殘餘物在庚烷(18L)中漿化約16小時。將固體藉由過濾收集，在氮氣下一次性過濾並用庚烷(3L)洗滌以產出2196g所希望的物質(NMR測定純度為90%，HPLC純度為99%)。將該批次的剩餘物 在空氣下過濾並用庚烷(3L)洗滌以產出1905g所希望的物質(NMR測定純度為88%，HPLC純度為99%)。將該等黃色固體合併用於進一步加工(4101g，3653g有活性，83%產率，HPLC純度為99%)。 On a larger scale, ethyl 6-bromo-1-[(4-methoxyphenyl) methyl] -4-pentoxyquinoline-3-carboxylate (5765 g, 13.85 mol) was filled to have Sulphur chloride (28.8L) in a container. An exotherm from 20 ° C to 26 ° C was observed. DMF (4.4 mL) was added, no exotherm was observed, and the batch was heated to 75 ° C and stirred for 17 hours. HPLC showed that 98.0% of the product had 1.3% starting material remaining. The reaction was concentrated in vacuo and the residue was azeotroped with toluene (25L). The resulting solid was then slurried in heptane (18.5L) for 2.5 hours, filtered and washed with heptane (3 x 4L). The solid was dried under vacuum at 35 ° C. to give 4077 g of the desired substance (93% crude yield). By HPLC, this substance contained about 5% of 6-bromo in addition to about 4% of the hydrolysate. 1-[(4-methoxyphenyl) methyl] -4-oxoquinoline-3-carboxylic acid ethyl ester (90% pure). The crude material (4077g) was returned to the container and reprocessed with sulfenyl chloride (14.5L) and DMF (2.2mL). The mixture was heated to 75 ° C for 40 hours. The thionyl chloride was removed in vacuo and the residue was azeotroped with toluene (10 L). At 20 ° C, the residue was slurried in heptane (18 L) for about 16 hours. The solid was collected by filtration, filtered at once under nitrogen and washed with heptane (3L) to yield 2196g of the desired material (purity by NMR was 90%, purity by HPLC was 99%). The remainder of the batch was filtered under air and washed with heptane (3L) to yield 1905g of the desired material (purity determined by NMR 88%, purity HPLC 99%). The yellow solids were combined for further processing (4101g, 3653g active, 83% yield, HPLC purity 99%).

中間體A6：6-溴-1-[(4-甲氧基苯基)甲基]-4-側氧基喹啉-3-甲酸乙酯Intermediate A6: 6-bromo-1-[(4-methoxyphenyl) methyl] -4-oxoquinoline-3-carboxylic acid ethyl ester

在環境溫度下，經2分鐘的時段將1,8-二氮雜二環[5.4.0]十一-7-烯(102mL，679.62mmol)滴加到在丙酮(1.2L)中的2-(5-溴-2-氟苯甲醯基)-3-[(4-甲氧基苯基)甲基胺基]丙-2-烯酸乙酯(296.5g，679.62mmol)中。將所得的溶液攪拌16小時然後藉由過濾去除固體並用三級丁基甲基醚洗滌，以得到呈淺黃色固體的所希望的物質(180g，64%)。NMR譜：1H NMR(400MHz,DMSO-d6)δ 1.30(3H,t),3.71(3H,s),4.25(2H,q),5.60(2H,s),6.90-6.95(2H,m),7.12-7.25(2H,m),7.67(1H,d),7.80-7.90(1H,m),8.30(1H,d),8.92(1H,s)。質譜：m/z(ES+)[M+H]+=418。 At ambient temperature, 1,8-diazabicyclo [5.4.0] undec-7-ene (102mL, 679.62mmol) was added dropwise to 2- in acetone (1.2L) over a period of 2 minutes (5-Bromo-2-fluorobenzyl) -3-[(4-methoxyphenyl) methylamino] prop-2-enoic acid ethyl ester (296.5 g, 679.62 mmol). The resulting solution was stirred for 16 hours and then the solid was removed by filtration and washed with tertiary butyl methyl ether to obtain the desired substance (180 g, 64%) as a pale yellow solid. NMR spectrum: 1 H NMR (400MHz, DMSO-d6) δ 1.30 (3H, t), 3.71 (3H, s), 4.25 (2H, q), 5.60 (2H, s), 6.90-6.95 (2H, m) , 7.12-7.25 (2H, m), 7.67 (1H, d), 7.80-7.90 (1H, m), 8.30 (1H, d), 8.92 (1H, s). Mass spectrum : m / z (ES +) [M + H] + = 418.

以更大的規模，在15℃下，將2-(5-溴-2-氟苯甲醯基)-3-[(4-甲氧基苯基)甲基胺基]丙-2-烯酸乙酯(8434g，(7730g假定有活性)，17.71mol)填裝至具有丙酮(23.2L)的容器中。經25分鐘添加1,8-二氮雜二環[5.4.0]十一-7-烯(2.8L，18.72mol)，隨著添加觀察到從18℃-23℃的放熱。約25分鐘後沈澱物形成並且 該批次持續放熱，1小時後達到37℃的最高值。將該反應在20℃下攪拌16.5小時，在此時HPLC指示起始物質的消耗和96.5%產物。藉由過濾，用三級丁基甲基醚(4 x 3.4L)洗滌來收集所得的沈澱物。然後將固體在真空下在40℃下乾燥，以給出6033g呈白色固體的所希望的物質(經3個步驟81.6%產率，HPLC純度為99.8%)。分析數據與針對先前批次獲得的是一致的。 On a larger scale, at 15 ° C, 2- (5-bromo-2-fluorobenzyl) -3-[(4-methoxyphenyl) methylamino] prop-2-ene Ethyl acid ester (8434g, (7730g assumed to be active), 17.71mol) was filled into a container with acetone (23.2L). 1,8-Diazabicyclo [5.4.0] undec-7-ene (2.8 L, 18.72 mol) was added over 25 minutes, and an exotherm from 18 ° C to 23 ° C was observed with the addition. A precipitate formed after about 25 minutes and the batch continued to exotherm, reaching a maximum value of 37 ° C after 1 hour. The reaction was stirred at 20 ° C for 16.5 hours, at which time HPLC indicated consumption of starting material and 96.5% product. The resulting precipitate was collected by filtration and washed with tertiary butyl methyl ether (4 x 3.4L). The solid was then dried under vacuum at 40 ° C to give 6033 g of the desired material as a white solid (81.6% yield over 3 steps, HPLC purity 99.8%). The analysis data is consistent with those obtained for the previous batch.

中間體A7：2-(5-溴-2-氟苯甲醯基)-3-[(4-甲氧基苯基)甲基胺基]丙-2-烯酸乙酯Intermediate A7: ethyl 2- (5-bromo-2-fluorobenzyl) -3-[(4-methoxyphenyl) methylamino] prop-2-enoate

在10℃下，經10分鐘的時段將(E)-3-(二甲基胺基)丙烯酸乙酯(98g，685.00mmol)分批添加至在甲苯(800mL)中的5-溴-2-氟苯甲醯氯(163g，685mmol)和DIPEA(120mL，685.00mmol)中。將所得的溶液在70℃下攪拌16小時然後允許冷卻。在環境溫度下，經20分鐘的時段將(4-甲氧基苯基)甲胺(94g，685mmol)添加至該混合物中。將所得的溶液攪拌3小時然後將該反應混合物用DCM(4L)稀釋，並用水(3 x 1L)洗滌。將有機相經Na2SO4乾燥，過濾並且蒸發以給出呈棕色油狀物的所希望的物質(300g，100%)，將其不進行進一步純化立刻用於隨後的反應中。質譜：m/z(ES+)[M+H]+=436。 At 10 ° C., (E) -3- (dimethylamino) ethyl acrylate (98 g, 685.00 mmol) was added portionwise to 5-bromo-2- in toluene (800 mL) over a 10-minute period Fluorobenzoyl chloride (163 g, 685 mmol) and DIPEA (120 mL, 685.00 mmol). The resulting solution was stirred at 70 ° C for 16 hours and then allowed to cool. At ambient temperature, (4-methoxyphenyl) methylamine (94 g, 685 mmol) was added to the mixture over a period of 20 minutes. The resulting solution was stirred for 3 hours and then the reaction mixture was diluted with DCM (4L) and washed with water (3 x 1L). The organic phase was dried over Na 2 SO 4 , filtered and evaporated to give the desired material (300 g, 100%) as a brown oil, which was used in the subsequent reaction immediately without further purification. Mass spectrum : m / z (ES +) [M + H] + = 436.

以更大的規模，將5-溴-2-氟苯甲醯氯(4318g，4205g有活性，17.71mol)填裝至容器中作為在甲苯(7.5L)中的溶液。 添加DIPEA(3150mL，18.08mol)，沒有觀察到放熱。經30分鐘分批添加3-(二甲基胺基)丙烯酸乙酯(2532g，17.71mol)，維持批次溫度<40℃。隨著30分鐘添加，注意到從21℃-24℃放熱，經1小時進一步緩慢升高至38℃。將該反應在20℃-30℃下攪拌16.5小時。經30分鐘分批添加4-甲氧基苄胺(2439g，17.78mol)，維持批次溫度<40℃。隨著添加觀察到25℃-30℃的放熱，藉由15℃的降低的夾套溫度提供冷卻。將該反應在20℃-30℃下攪拌4小時，這之後HPLC指示93.2%的所希望的物質。將批次分離用於後處理，將該混合物的每一半用DCM(28.6L)稀釋並用水(3 x 7.8L)洗滌。將有機物經MgSO4(約550g)乾燥並過濾，用DCM(4L)洗滌。然後將合併的有機物濃縮以給出8444g呈油狀物的所希望的物質(8434g，106%產率，HPLC純度為94.7%)。分析數據與針對先前批次獲得的是一致的。 On a larger scale, 5-bromo-2-fluorobenzoyl chloride (4318 g, 4205 g active, 17.71 mol) was filled into a container as a solution in toluene (7.5 L). DIPEA (3150 mL, 18.08 mol) was added, and no exotherm was observed. Ethyl 3- (dimethylamino) acrylate (2532 g, 17.71 mol) was added in portions over 30 minutes, maintaining the batch temperature <40 ° C. With the addition of 30 minutes, an exotherm was noticed from 21 ° C to 24 ° C, and the temperature gradually increased to 38 ° C over 1 hour. The reaction was stirred at 20-30 ° C for 16.5 hours. 4-Methoxybenzylamine (2439 g, 17.78 mol) was added in portions over 30 minutes, maintaining the batch temperature <40 ° C. An exotherm of 25 ° C-30 ° C was observed with the addition, cooling was provided by a reduced jacket temperature of 15 ° C. The reaction was stirred at 20-30 ° C for 4 hours, after which HPLC indicated 93.2% of the desired material. The batch was separated for workup, each half of the mixture was diluted with DCM (28.6L) and washed with water (3 x 7.8L). The organics over MgSO 4 (about 550g) was dried and filtered, washed with DCM (4L). The combined organics were then concentrated to give 8444 g of the desired material as an oil (8434 g, 106% yield, HPLC purity 94.7%). The analysis data is consistent with those obtained for the previous batch.

中間體A8：5-溴-2-氟苯甲醯氯Intermediate A8: 5-bromo-2-fluorobenzoyl chloride

在環境溫度下，經1小時的時間段將亞硫醯氯(75.0mL，1027.36mmol)滴加至在甲苯(1.2L)和DMF(12mL)中的5-溴-2-氟苯甲酸(150g，684.91mmoL)中。將所得的混合物在70℃下攪拌16小時然後允許將該混合物冷卻並在真空中濃縮，以得到呈淺黃色油狀物的所希望的物質(160g，98%)，將其不進行進一步純化而使用。NMR譜：1H NMR(400MHz,DMSO-d6)δ 7.26-7.31(1H,m),7.83(1H,dd),8.02(1H,d)。 At ambient temperature, thionyl chloride (75.0 mL, 1027.36 mmol) was added dropwise to 5-bromo-2-fluorobenzoic acid (150 g) in toluene (1.2 L) and DMF (12 mL) over a period of 1 hour , 684.91mmoL). The resulting mixture was stirred at 70 ° C for 16 hours and then the mixture was allowed to cool and concentrated in vacuo to obtain the desired material (160 g, 98%) as a pale yellow oil, which was used without further purification. use. NMR spectrum: 1 H NMR (400 MHz, DMSO-d6) δ 7.26-7.31 (1H, m), 7.83 (1H, dd), 8.02 (1H, d).

以更大的規模，在20℃下，將3-溴-6-氟苯甲酸(3888g，17.75mol)填裝至容器中，隨後填裝甲苯(29.2L)。添加亞硫醯氯(1950mL，26.88mol)，隨後添加DMF(310mL)，沒有觀察到放熱。將該混合物加熱至65℃-75℃(約45℃之上獲得溶液)，沒有觀察到放熱和輕微氣體逸出。將該反應在此溫度下攪拌40小時，此時HPLC分析顯示87.6%產物，3.4%起始物質。將該反應在真空中濃縮並與甲苯(18L)共沸，以給出4328g所希望的物質(103%產率，HPLC純度為87.3%)。 On a larger scale, at 20 ° C., 3-bromo-6-fluorobenzoic acid (3888 g, 17.75 mol) was filled into a container, followed by toluene (29.2 L). Thionyl chloride (1950 mL, 26.88 mol) was added, followed by DMF (310 mL), and no exotherm was observed. The mixture was heated to 65 ° C-75 ° C (solution obtained above about 45 ° C), no exotherm and slight gas evolution was observed. The reaction was stirred at this temperature for 40 hours, at which time HPLC analysis showed 87.6% product, 3.4% starting material. The reaction was concentrated in vacuo and azeotroped with toluene (18 L) to give 4328 g of the desired material (103% yield, HPLC purity 87.3%).

中間體C5：6-溴-4-氯-7-氟喹啉-3-甲酸乙酯Intermediate C5: ethyl 6-bromo-4-chloro-7-fluoroquinoline-3-carboxylate

在10℃下，在惰性氣氛下，將DMF(0.535mL，6.91mmol)添加至在亞硫醯氯(600mL)中的6-溴-7-氟-1-[(4-甲氧基苯基)甲基]-4-側氧基-喹啉-3-甲酸乙酯(200g，460.56mmol)中並將所得的混合物在70℃下攪拌3小時。將該混合物蒸發至乾燥，並且將殘餘物與甲苯(300mL)共沸，以得到粗產物。將該粗產物藉由從己烷結晶進行純化，以得到呈白色固體的所希望的物質(147g，96%)。NMR譜： 1H NMR(400MHz,CDCl3)δ 1.49(3H,t),4.51-4.56(2H,m),7.91(1H,d),8.71(1H,d),9.26(1H,s)。質譜：m/z(ES+)[M+H]+=334。 At 10 ° C, under an inert atmosphere, DMF (0.535 mL, 6.91 mmol) was added to 6-bromo-7-fluoro-1-[(4-methoxyphenyl) in thiosulfonyl chloride (600 mL) ) Methyl] -4-oxo-quinoline-3-carboxylic acid ethyl ester (200 g, 460.56 mmol) and the resulting mixture was stirred at 70 ° C. for 3 hours. The mixture was evaporated to dryness, and the residue was azeotroped with toluene (300 mL) to obtain a crude product. The crude product was purified by crystallization from hexane to obtain the desired substance as a white solid (147 g, 96%). NMR spectrum: 1 H NMR (400 MHz, CDCl 3 ) δ 1.49 (3H, t), 4.51-4.56 (2H, m), 7.91 (1H, d), 8.71 (1H, d), 9.26 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 334.

中間體C6：6-溴-7-氟-1-[(4-甲氧基苯基)甲基]-4-側氧基-喹啉-3-甲酸乙酯Intermediate C6: 6-bromo-7-fluoro-1-[(4-methoxyphenyl) methyl] -4-oxo-quinoline-3-carboxylic acid ethyl ester

在10℃下，經5分鐘的時段，在惰性氣氛下，將1,8-二氮雜二環[5.4.0]十一-7-烯(76mL，506.32mmol)緩慢添加到在丙酮(800mL)中的2-(5-溴-2,4-二氟-苯甲醯基)-3-[(4-甲氧基苯基)甲基胺基]丙-2-烯酸乙酯(230g，506.32mmol)中，並且將所得的混合物在環境溫度下攪拌16小時。藉由過濾收集沈澱物，用Et2O(3 x 500mL)洗滌並在真空下乾燥，以得到呈白色固體的所希望的物質(166g，75%)。NMR譜： 1H NMR(400MHz,DMSO-d6)δ 1.29(3H,t),3.72(3H,s),4.22-4.27(21H,m),5.57(2H,s),6.92-6.95(2H,m),7.24(2H,d),7.79(1H,d),8.40(1H,d),8.89(1H,s)。質譜：m/z(ES+)[M+H]+=434。 At 10 ° C, 1,8-diazabicyclo [5.4.0] undec-7-ene (76mL, 506.32mmol) was slowly added to acetone (800mL) under an inert atmosphere over a period of 5 minutes ) 2- (5-Bromo-2,4-difluoro-benzoyl) -3-[(4-methoxyphenyl) methylamino] prop-2-enoic acid ethyl ester (230g , 506.32 mmol), and the resulting mixture was stirred at ambient temperature for 16 hours. The precipitate was collected by filtration, washed with Et 2 O (3 x 500 mL) and dried under vacuum to obtain the desired material as a white solid (166 g, 75%). NMR spectrum: 1 H NMR (400 MHz, DMSO-d6) δ 1.29 (3H, t), 3.72 (3H, s), 4.22-4.27 (21H, m), 5.57 (2H, s), 6.92-6.95 (2H, m), 7.24 (2H, d), 7.79 (1H, d), 8.40 (1H, d), 8.89 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 434.

中間體C7：2-(5-溴-2,4-二氟-苯甲醯基)-3-[(4-甲氧基苯基)甲基胺基]丙-2-烯酸乙酯Intermediate C7: 2- (5-Bromo-2,4-difluoro-benzoyl) -3-[(4-methoxyphenyl) methylamino] prop-2-enoic acid ethyl ester

在環境溫度下，在惰性氣氛下，將(E)-3-(二甲基胺基)丙烯酸乙酯(80mL，555.50mmol)滴加至DIPEA(132mL，757.50mmol)和5-溴-2,4-二氟-苯甲醯氯(129g，505.00mmol)在甲苯(600mL)中的混合物中。將所得的溶液在70℃下攪拌17小時然後允許冷卻。將(4-甲氧基苯基)甲胺(66.0mL，505.29mmol) 分批添加至該混合物中並將該反應在環境溫度下攪拌3小時。將該反應混合物用DCM(2L)稀釋，順序地用水(4 x 200mL)、飽和鹽水(300mL)洗滌，將有機層經Na2SO4乾燥，過濾並且蒸發以得到呈淺棕色固體的所希望的物質(230g，100%)，將其不進行進一步純化而用於下一步。NMR譜： 1H NMR(400MHz,CDCl3)δ 1.09(3H,t),3.82(3H,s),4.00-4.10(2H,m),4.55(2H,t),6.84-6.96(3H,m),7.20-7.29(2H,m),7.55(1H,d),8.18(1H,t)質譜：m/z(ES+)[M+H]+=454。 At ambient temperature, under an inert atmosphere, (E) -3- (dimethylamino) ethyl acrylate (80 mL, 555.50 mmol) was added dropwise to DIPEA (132 mL, 757.50 mmol) and 5-bromo-2, A mixture of 4-difluoro-benzoyl chloride (129 g, 505.00 mmol) in toluene (600 mL). The resulting solution was stirred at 70 ° C for 17 hours and then allowed to cool. (4-Methoxyphenyl) methylamine (66.0 mL, 505.29 mmol) was added to the mixture in portions and the reaction was stirred at ambient temperature for 3 hours. The reaction mixture was diluted with DCM (2 L), washed sequentially with water (4 x 200 mL), saturated brine (300 mL), the organic layer was dried over Na 2 SO 4 , filtered and evaporated to give the desired as a light brown solid The material (230g, 100%) was used in the next step without further purification. NMR spectrum: 1 H NMR (400 MHz, CDCl 3 ) δ 1.09 (3H, t), 3.82 (3H, s), 4.00-4.10 (2H, m), 4.55 (2H, t), 6.84-6.96 (3H, m ), 7.20-7.29 (2H, m), 7.55 (1H, d), 8.18 (1H, t) Mass Spectrum : m / z (ES +) [M + H] + = 454.

中間體C8：5-溴-2,4-二氟-苯甲醯氯Intermediate C8: 5-bromo-2,4-difluoro-benzoyl chloride

在15℃下，經5分鐘的時間段，在惰性氣氛下，將亞硫醯氯(55.4mL，759.50mmol)分批添加至DMF(7.84mL，101.27mmol)和5-溴-2,4-二氟苯甲酸(120g，506.33mmol)在甲苯(600mL)中的混合物中。將所得的混合物在70℃下攪拌4小時然後蒸發至乾燥並將殘餘物與甲苯共沸，以得到呈棕色油狀物的所希望的物質(129g，100%)，將其不進行純化直接用於下一步。NMR譜： 1H NMR(400MHz,CDCl3)δ 7.04-7.09(1H,m),8.34-8.42(1H,m)。 At 15 ° C, for 5 minutes, under an inert atmosphere, thionyl chloride (55.4 mL, 759.50 mmol) was added portionwise to DMF (7.84 mL, 101.27 mmol) and 5-bromo-2,4- Difluorobenzoic acid (120 g, 506.33 mmol) in a mixture of toluene (600 mL). The resulting mixture was stirred at 70 ° C for 4 hours and then evaporated to dryness and the residue was azeotroped with toluene to obtain the desired substance as a brown oil (129g, 100%), which was used without purification In the next step. NMR spectrum: 1 H NMR (400 MHz, CDCl 3 ) δ 7.04-7.09 (1H, m), 8.34-8.42 (1H, m).

如下製備中間體外消旋-8-溴-7-氟-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮：中間體F2：外消旋-8-溴-7-氟-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮 The intermediate racemic-8-bromo-7-fluoro-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5-c] quinoline-2 (3H)-was prepared as follows Ketone: intermediate F2: racemic-8-bromo-7-fluoro-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5-c] quinoline-2 ( 3H) -ketone

將三氯異氰尿酸(213mg，0.92mmol)一次性添加到在MeOH(10mL)中的6-溴-7-氟-4-((四氫-2H-哌喃-3-基)胺基)喹啉-3-甲醯胺(843mg，2.29mmol)和1,8-二氮雜二環[5.4.0]十一-7-烯(0.629mL，4.58mmol)中。將所得的混合物在室溫下攪拌70小時。該反應不完全，並且添加另外的三氯異氰尿酸(213mg，0.92mmol)和1,8-二氮雜二環[5.4.0]十一-7-烯(0.629mL，4.58mmol)，並且將混合物在室溫下再攪拌3小時。將所得的混合物蒸發至乾燥，並且將殘餘物藉由FCC進行純化，洗提梯度為在DCM中的0至10% MeOH。將純級分蒸發至乾燥，以得到呈奶油色固體的外消旋-8-溴-7-氟-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮(585mg，69.8%)。NMR譜：1H NMR(400MHz,DMSO-d6)δ 1.78-1.88(2H,m),2.10(1H,d),2.62-2.73(1H,m),3.40(1H,td),3.88-4.04(1H,m),4.02-4.12(1H,m),4.19(1H,t),4.72-4.80(1H,m),7.94(1H,d),8.48(1H,d),8.69(1H,s),11.62(1H,s)。質譜：m/z(ES+)[M+H]+=366。 Trichloroisocyanuric acid (213 mg, 0.92 mmol) was added in one portion to 6-bromo-7-fluoro-4-((tetrahydro-2H-piperan-3-yl) amino) in MeOH (10 mL) Quinoline-3-carboxamide (843 mg, 2.29 mmol) and 1,8-diazabicyclo [5.4.0] undec-7-ene (0.629 mL, 4.58 mmol). The resulting mixture was stirred at room temperature for 70 hours. The reaction was incomplete, and additional trichloroisocyanuric acid (213 mg, 0.92 mmol) and 1,8-diazabicyclo [5.4.0] undec-7-ene (0.629 mL, 4.58 mmol) were added, and The mixture was stirred at room temperature for another 3 hours. The resulting mixture was evaporated to dryness, and the residue was purified by FCC with an elution gradient of 0 to 10% MeOH in DCM. The pure fractions were evaporated to dryness to give racemic-8-bromo-7-fluoro-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4, 5-c] Quinolin-2 (3H) -one (585 mg, 69.8%). NMR spectrum : 1 H NMR (400 MHz, DMSO-d 6 ) δ 1.78-1.88 (2H, m), 2.10 (1H, d), 2.62-2.73 (1H, m), 3.40 (1H, td), 3.88-4.04 (1H, m), 4.02-4.12 (1H, m), 4.19 (1H, t), 4.72-4.80 (1H, m), 7.94 (1H, d), 8.48 (1H, d), 8.69 (1H, s ), 11.62 (1H, s). Mass spectrum : m / z (ES +) [M + H] + = 366.

中間體F3：外消旋-6-溴-7-氟-4-((四氫-2H-哌喃-3-基)胺基)喹啉-3-甲醯胺Intermediate F3: Racemic-6-bromo-7-fluoro-4-((tetrahydro-2H-piperan-3-yl) amino) quinoline-3-carboxamide

將6-溴-4-氯-7-氟喹啉-3-甲醯胺(1g，3.29mmol)、 四氫-2H-哌喃-3-胺(0.350g，3.46mmol)和DIPEA(1.435mL，8.24mmol)懸浮於DMA(10mL)中，並將所得的混合物在100℃下加熱過夜。將該反應混合物用水稀釋，並且藉由過濾收集沈澱物，用水(10mL)洗滌並在真空下乾燥，以得到呈米色固體的外消旋-6-溴-7-氟-4-((四氫-2H-哌喃-3-基)吡啶-2-基)胺基)喹啉-3-甲醯胺(0.843g，69.5%)，將其不經進一步純化而使用。NMR譜：1H NMR(400MHz,DMSO-d6)δ 1.47-1.55(1H,m),1.60-1.72(2H,m),2.02(1H,d),3.33-3.48(2H,m),3.68(1H,dd),3.87(1H,dd),3.91-3.97(1H,m),7.56(1H,s),7.71(1H,d),8.11(1H,s),8.20(1H,d),8.62(2H,d)。質譜：m/z(ES+)[M+H]+=368。 Combine 6-bromo-4-chloro-7-fluoroquinoline-3-carboxamide (1g, 3.29mmol), tetrahydro-2H-piperan-3-amine (0.350g, 3.46mmol) and DIPEA (1.435mL) , 8.24 mmol) was suspended in DMA (10 mL), and the resulting mixture was heated at 100 ° C. overnight. The reaction mixture was diluted with water, and the precipitate was collected by filtration, washed with water (10 mL) and dried under vacuum to give racemic-6-bromo-7-fluoro-4-((tetrahydro -2H-piperan-3-yl) pyridin-2-yl) amino) quinoline-3-carboxamide (0.843 g, 69.5%), which was used without further purification. NMR spectrum : 1 H NMR (400 MHz, DMSO-d 6 ) δ 1.47-1.55 (1H, m), 1.60-1.72 (2H, m), 2.02 (1H, d), 3.33-3.48 (2H, m), 3.68 (1H, dd), 3.87 (1H, dd), 3.91-3.97 (1H, m), 7.56 (1H, s), 7.71 (1H, d), 8.11 (1H, s), 8.20 (1H, d), 8.62 (2H, d). Mass spectrum: m / z (ES +) [M + H] + = 368.

中間體F4：6-溴-4-氯-7-氟-喹啉-3-甲醯胺Intermediate F4: 6-bromo-4-chloro-7-fluoro-quinoline-3-carboxamide

將DMF(0.5mL)添加到6-溴-7-氟-4-側氧基-1H-喹啉-3-甲酸(22.5g，78.66mmol)在亞硫醯氯(140g，1179.85mmol)中的攪拌懸浮液中，並且將混合物加熱至回流2小時。允許將反應冷卻，在真空中濃縮，並且將殘餘物與甲苯共沸兩次，以得到黃色固體。在0℃下，將該固體分批添加至氫氧化銨的溶液(147mL，1179.85mmol)中。將白色懸浮液攪拌15分鐘然後將固體過濾，用水洗滌並在真空下乾燥，以得到呈白色粉末狀的所希望的物質(23.80g，100%)。NMR譜： 1H NMR(400MHz,DMSO-d6)δ 8.92(1H,s),8.59(1H,d),8.21(1H,s),8.09(1H,d),7.98(1H,s)。質譜：m/z(ES+)[M+H]+=304.8 DMF (0.5 mL) was added to the 6-bromo-7-fluoro-4-oxo-1H-quinoline-3-carboxylic acid (22.5 g, 78.66 mmol) in thionyl chloride (140 g, 1179.85 mmol) The suspension was stirred and the mixture was heated to reflux for 2 hours. The reaction was allowed to cool, concentrated in vacuo, and the residue was azeotroped twice with toluene to give a yellow solid. At 0 ° C., the solid was added portionwise to a solution of ammonium hydroxide (147 mL, 1179.85 mmol). The white suspension was stirred for 15 minutes and then the solid was filtered, washed with water and dried under vacuum to obtain the desired substance as a white powder (23.80 g, 100%). NMR spectrum: 1 H NMR (400 MHz, DMSO-d6) δ 8.92 (1H, s), 8.59 (1H, d), 8.21 (1H, s), 8.09 (1H, d), 7.98 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 304.8

中間體F5：6-溴-7-氟-4-側氧基-1H-喹啉-3-甲酸Intermediate F5: 6-bromo-7-fluoro-4-oxo-1H-quinoline-3-carboxylic acid

在環境溫度下，將氫氧化鈉(18.34g，458.44mmol)在水(100mL)中的溶液添加至6-溴-7-氟-4-側氧基-1H-喹啉-3-甲酸乙酯(28.8g，91.69mmol)在EtOH(500mL)中的攪拌懸浮液中。然後將該反應混合物在75℃下攪拌2小時，允許冷卻並使用2N鹽酸將pH調節至4。藉由過濾收集沈澱物，用水洗滌並在真空下乾燥，以得到呈白色粉末狀的所希望的物質(23.30g，89%)。NMR譜： 1H NMR(400MHz,DMSO-d6)δ 14.78(1H,s),13.45(1H,s),8.93(1H,s),8.46(1H,d),7.70(1H,d)。質譜：m/z(ES+)[M+H]+=287.8 At ambient temperature, add a solution of sodium hydroxide (18.34 g, 458.44 mmol) in water (100 mL) to ethyl 6-bromo-7-fluoro-4-oxo-1H-quinoline-3-carboxylate (28.8 g, 91.69 mmol) in a stirred suspension in EtOH (500 mL). The reaction mixture was then stirred at 75 ° C for 2 hours, allowed to cool and adjusted to pH 4 with 2N hydrochloric acid. The precipitate was collected by filtration, washed with water and dried under vacuum to obtain the desired substance as a white powder (23.30 g, 89%). NMR spectrum: 1 H NMR (400 MHz, DMSO-d6) δ 14.78 (1H, s), 13.45 (1H, s), 8.93 (1H, s), 8.46 (1H, d), 7.70 (1H, d). Mass spectrum: m / z (ES +) [M + H] + = 287.8

中間體F6：6-溴-7-氟-4-側氧基-1H-喹啉-3-甲酸乙酯Intermediate F6: 6-Bromo-7-fluoro-4-oxo-1H-quinoline-3-carboxylic acid ethyl ester

將2-[(4-溴-3-氟-苯胺基)亞甲基]丙二酸二乙酯(90g，249.88mmol)在二苯醚(600mL，3.79mol)中的溶液在240℃下攪拌2.5小時。允許將該混合物冷卻至70℃，將該等固體藉由過濾收集並在真空烘箱乾燥，以得到呈白色固體的所希望的物質(50g，64%)，將其不進行進一步純化而使用。NMR譜： 1H NMR(500MHz,DMSO-d6,(100℃))δ 1.26-1.33(3H,m),4.25(2H,q),7.52(1H,d),8.37(1H,d),8.48(1H,s),12.05(1H,s)。質譜：m/z(ES+)[M+H]+=314 A solution of 2-[(4-bromo-3-fluoro-anilino) methylene] malonic acid diethyl ester (90 g, 249.88 mmol) in diphenyl ether (600 mL, 3.79 mol) was stirred at 240 ° C 2.5 hours. The mixture was allowed to cool to 70 ° C, and the solids were collected by filtration and dried in a vacuum oven to obtain the desired material (50 g, 64%) as a white solid, which was used without further purification. NMR spectrum: 1 H NMR (500MHz, DMSO-d6, (100 ° C)) δ 1.26-1.33 (3H, m), 4.25 (2H, q), 7.52 (1H, d), 8.37 (1H, d), 8.48 (1H, s), 12.05 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 314

中間體F7：2-[(4-溴-3-氟-苯胺基)亞甲基]丙二酸二乙酯Intermediate F7: 2-[(4-bromo-3-fluoro-anilino) methylene] malonic acid diethyl ester

將4-溴-3-氟苯胺(56.6g，297.87mmol)和1,3-2-(乙氧基亞甲基)丙二酸二乙酯(72.45g，335.06mmol)在EtOH(560mL)中的溶液在80℃下攪拌4小時。允許將該反應混合物冷卻，將該等固體藉由過濾收集並在烘箱中乾燥，以得到呈灰白色固體的所希望的物質(90g，84%)，將其不進行進一步純化而使用。NMR譜： 1H NMR(400MHz,DMSO-d6)δ 1.26(6H,q),4.14(2H,q),4.22(2H,q),7.18-7.25(1H,m),7.57(1H,dd),7.64-7.7(1H,m),8.33(1H,d),10.62(1H,d)。質譜：m/z(ES+)[M+H]+=360。 4-Bromo-3-fluoroaniline (56.6 g, 297.87 mmol) and diethyl 1,3-2- (ethoxymethylene) malonate (72.45 g, 335.06 mmol) in EtOH (560 mL) The solution was stirred at 80 ° C for 4 hours. The reaction mixture was allowed to cool, and the solids were collected by filtration and dried in an oven to obtain the desired substance (90 g, 84%) as an off-white solid, which was used without further purification. NMR spectrum: 1 H NMR (400MHz, DMSO-d6) δ 1.26 (6H, q), 4.14 (2H, q), 4.22 (2H, q), 7.18-7.25 (1H, m), 7.57 (1H, dd) , 7.64-7.7 (1H, m), 8.33 (1H, d), 10.62 (1H, d). Mass spectrum: m / z (ES +) [M + H] + = 360.

實例9 Example 9

外消旋-1-(3,3-二甲基四氫-2H-哌喃-4-基)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮Racemic-1- (3,3-dimethyltetrahydro-2H-piperan-4-yl) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one

將二氯雙(二-三級丁基(3-磺丙基)磷鎓基)鈀酸鹽(II)(在水中的0.05M)(31.4mL，1.57mmol)添加到2-(甲氧基甲基)-5-(4,4,5,5-四甲基-1,3,2-二氧雜環戊硼烷-2-基)吡啶(8.59g，34.50mmol)、外消旋-8-溴-1-(3,3-二甲基四氫-2H-哌喃-4-基)-3-甲基-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮(12.24g，31.36mmol)以 及2M K2CO3溶液(47.0mL，94.09mmol)在1,4-二(188mL)和水(47.0mL)中的脫氣混合物中。將該反應加熱至80℃持續4小時。將該反應混合物濃縮並用DCM(250mL)稀釋，並且用水(200mL)洗滌。將有機層用相分離柱乾燥並蒸發，以得到粗產物。將該米色固體在MeCN中加熱，並冷卻至室溫。在真空下過濾固體，以得到呈米色固體的外消旋-1-(3,3-二甲基四氫-2H-哌喃-4-基)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮(12g，88%)。NMR譜： 1H NMR(500MHz,CDCl3)δ 0.94(3H,s),1.35(3H,s),1.73(1H,d),3.31-3.41(1H,m),3.57(3H,s),3.60(3H,s),3.63-3.82(3H,m),4.26(1H,dd),4.69(2H,s),5.02(1H,dd),7.51-7.64(1H,m),7.83(1H,dd),8.02(1H,dd),8.27(1H,d),8.60(1H,d),8.74(1H,s),8.93(1H,dt)。質譜：m/z(ES+)[M+H]+=433。 Add dichlorobis (di-tertiary butyl (3-sulfopropyl) phosphonium) palladium (II) (0.05M in water) (31.4mL, 1.57mmol) to 2- (methoxy) Methyl) -5- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) pyridine (8.59g, 34.50mmol), racemic- 8-Bromo-1- (3,3-dimethyltetrahydro-2H-piperan-4-yl) -3-methyl-1,3-dihydro-2H-imidazo [4,5-c] Quinolin-2-one (12.24g, 31.36mmol) and 2M K 2 CO 3 solution (47.0mL, 94.09mmol) in 1,4-di (188 mL) and degassed mixture in water (47.0 mL). The reaction was heated to 80 ° C for 4 hours. The reaction mixture was concentrated and diluted with DCM (250 mL), and washed with water (200 mL). The organic layer was dried with a phase separation column and evaporated to obtain a crude product. The beige solid was heated in MeCN and cooled to room temperature. The solid was filtered under vacuum to give racemic-1- (3,3-dimethyltetrahydro-2H-piperan-4-yl) -8- (6- (methoxymethyl) as a beige solid ) Pyridin-3-yl) -3-methyl-1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one (12 g, 88%). NMR spectrum: 1 H NMR (500 MHz, CDCl 3 ) δ 0.94 (3H, s), 1.35 (3H, s), 1.73 (1H, d), 3.31-3.41 (1H, m), 3.57 (3H, s), 3.60 (3H, s), 3.63-3.82 (3H, m), 4.26 (1H, dd), 4.69 (2H, s), 5.02 (1H, dd), 7.51-7.64 (1H, m), 7.83 (1H, dd), 8.02 (1H, dd), 8.27 (1H, d), 8.60 (1H, d), 8.74 (1H, s), 8.93 (1H, dt). Mass spectrum: m / z (ES +) [M + H] + = 433.

分離上述外消旋混合物，以給出單獨的鏡像異構物組分，如下所述的實例10和11。已使用虛擬圓二色性法(VCD)分配了該等組分的立體化學。 The above racemic mixture was separated to give individual enantiomer components, as described in Examples 10 and 11 below . The stereochemistry of these components has been assigned using the virtual circular dichroism method (VCD).

將1-(3,3-二甲基四氫-2H-哌喃-4-基)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1H-咪唑并[4,5-c]喹啉-2(3H)-酮的外消旋混合物藉由製備型手性HPLC(Chiral Technologies IC柱，20μm二氧化矽，50mm直徑，250mm長度)進行分離，用在EtOH(用三乙胺改性)中的85%三級丁基甲基醚作為洗提液進行等度洗提，以得到呈固體的第一洗提產物實例10(314mg，30%)和呈固體的第二洗提產物實例11(331mg，32%)。 1- (3,3-Dimethyltetrahydro-2H-piperan-4-yl) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1H- The racemic mixture of imidazo [4,5-c] quinolin-2 (3H) -one was separated by preparative chiral HPLC (Chiral Technologies IC column, 20 μm silica, 50 mm diameter, 250 mm length), Isocratic elution was carried out with 85% tertiary butyl methyl ether in EtOH (modified with triethylamine) as the eluent to obtain the first solid elution product Example 10 (314 mg, 30%) and Example 11 of solid second elution product (331 mg, 32%).

實例10 Example 10

異構物1：(R)-1-(3,3-二甲基四氫-2H-哌喃-4-基)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮Isomer 1: ( R ) -1- (3,3-dimethyltetrahydro-2H-piperan-4-yl) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one

異構物1：NMR譜： 1H NMR(500MHz,DMSO-d6)δ 0.78(3H,s),1.05(1H,t),1.17(3H,d),1.64-1.82(1H,m),3.42(4H,s),3.50(4H,s),3.62-3.72(1H,m),4.06(1H,dd),4.59(2H,s),5.13(1H,dd),7.60(1H,dd),7.98(1H,dd),8.16(1H,d),8.26(1H,dd),8.74(1H,d),8.91(1H,s),9.00(1H,dd)。質譜：m/z(ES+)[M+H]+=433。 Isomer 1: NMR spectrum: 1 H NMR (500 MHz, DMSO-d6) δ 0.78 (3H, s), 1.05 (1H, t), 1.17 (3H, d), 1.64-1.82 (1H, m), 3.42 (4H, s), 3.50 (4H, s), 3.62-3.72 (1H, m), 4.06 (1H, dd), 4.59 (2H, s), 5.13 (1H, dd), 7.60 (1H, dd), 7.98 (1H, dd), 8.16 (1H, d), 8.26 (1H, dd), 8.74 (1H, d), 8.91 (1H, s), 9.00 (1H, dd). Mass spectrum: m / z (ES +) [M + H] + = 433.

實例11 Example 11

異構物2：(S)-1-(3,3-二甲基四氫-2H-哌喃-4-基)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮Isomer 2: ( S ) -1- (3,3-dimethyltetrahydro-2H-piperan-4-yl) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one

異構物2：NMR譜： 1H NMR(500MHz,DMSO d-6)δ 0.78(3H,s),0.95-1.12(1H,m),1.16(3H,s),1.68-1.81(1H,m),3.42(4H,m),3.50(4H,m),3.62-3.7(1H,m),4.07(1H,s),4.59(2H,s),5.13(1H,dd),7.60(1H,dd),7.98(1H,dd),8.16(1H,d),8.26(1H,dd),8.74(1H,d),8.91(1H,s),9.00(1H,dd)。質譜：m/z(ES+)[M+H]+=433。 Isomer 2: NMR spectrum: 1 H NMR (500 MHz, DMSO d-6) δ 0.78 (3H, s), 0.95-1.12 (1H, m), 1.16 (3H, s), 1.68-1.81 (1H, m ), 3.42 (4H, m), 3.50 (4H, m), 3.62-3.7 (1H, m), 4.07 (1H, s), 4.59 (2H, s), 5.13 (1H, dd), 7.60 (1H, dd), 7.98 (1H, dd), 8.16 (1H, d), 8.26 (1H, dd), 8.74 (1H, d), 8.91 (1H, s), 9.00 (1H, dd). Mass spectrum: m / z (ES +) [M + H] + = 433.

使用虛擬圓二色性法(VCD)分配立體化學。 Use virtual circular dichroism (VCD) to assign stereochemistry.

實驗：將灰白色固體樣品溶解於CDCl3中。將溶液轉移到0.100mm BaF2細胞中，並在裝備有雙源和雙光彈性調製器的BioTools ChiralIR儀器中分別獲得六個小時的VCD光譜。解析度為4cm-1。 Experimental: The samples were dissolved in an off-white solid in CDCl 3. The solution was transferred to 0.100 mm BaF 2 cells, and VCD spectra were obtained for six hours in a BioTools ChiralIR instrument equipped with a dual source and dual photoelastic modulator, respectively. The resolution is 4cm -1 .

電腦光譜類比：對S-鏡像異構物進行蒙特卡羅(Monte Carlo)分子力學對低能量幾何尺寸的搜索。使用Maestro圖形介面(薛定諤有限公司(Schrödinger Inc.))中的宏觀模型(MacroModel)來產生33個構象異構物的起始座標。將在5kcal/莫耳的最低能量構象異構物內的所有構象異構物用作Gaussian09內密度泛函理論(DFT)最小化的起點。在B3LYP/6-31G*水平的理論下，確定優化的結構、諧振動頻率/強度、VCD旋轉強度以及STP處的自由能(包括零點能量)。在計算總體超過10%的總體下，發現四個獨特的構象。該等顯示出四氫哌喃環的相同構象，但不同於吡啶環的取向和扭轉。 Computer spectroscopy analogy: search for low-energy geometries of S-image isomers by Monte Carlo molecular mechanics. A macro model in the Maestro graphical interface (Schrödinger Inc.) was used to generate the starting coordinates of 33 conformers. All conformational isomers within the lowest energy conformational isomer at 5 kcal / mole were used as the starting point for minimization of density functional theory (DFT) in Gaussian09. Under the theory of B3LYP / 6-31G * level, the optimized structure, resonance frequency / strength, VCD rotation strength and free energy (including zero point energy) at STP are determined. After calculating the total population of more than 10%, four unique conformations were found. These show the same conformation of the tetrahydropyran ring, but differ from the orientation and twist of the pyridine ring.

計算光譜和實驗光譜之間的擬合：使用內部程序產生紅外光譜和VCD光譜的模擬，以將Lorentzian線形(12cm-1線寬)擬合到計算的光譜，從而允許模擬光譜和實驗光譜之間的直接比較。低於1200cm-1的帶在很大程度上獨立於構象(並且因此反映在感興趣的中心處的絕對構型)。在1400cm-1和1300cm-1之間的帶對吡啶和稠合芳環的扭轉的符號(非鏡像物構象)係敏感的。(R-鏡像物的模擬光譜藉由S-鏡像物光譜的反轉獲得。)在針對異構物2(實例11)的計算的S-鏡像物和實驗光譜之間以及針對異構物 1(實例10)的計算的R-鏡像物和實驗光譜之間存在極好的一致性。在1400cm-1至1300cm-1的區域中的擬合也是合理的，這表明所計算的構象能量也是合理的。 Fitting between the calculated spectrum and the experimental spectrum: using an internal program to generate simulations of the infrared and VCD spectra to fit the Lorentzian line shape (12cm -1 line width) to the calculated spectrum, thereby allowing the simulation and experimental spectra Direct comparison. Bands below 1200 cm -1 are largely independent of conformation (and therefore reflect the absolute configuration at the center of interest). The bands between 1400 cm -1 and 1300 cm -1 are sensitive to the twisted symbols (non-mirror conformation) of pyridine and fused aromatic rings. (The simulated spectrum of the R-mirror is obtained by reversing the S-mirror spectrum.) Between the calculated S-mirror for isomer 2 ( Example 11 ) and the experimental spectrum and for isomer 1 ( Example 10 ) There is excellent agreement between the calculated R-mirror and the experimental spectrum. The fitting in the region of 1400 cm -1 to 1300 cm -1 is also reasonable, which indicates that the calculated conformational energy is also reasonable.

外消旋-8-溴-1-(3,3-二甲基四氫-2H-哌喃-4-基)-3-甲基-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮 Racemic-8-bromo-1- (3,3-dimethyltetrahydro-2H-piperan-4-yl) -3-methyl-1,3-dihydro-2H-imidazo [4, 5-c] quinolin-2-one

在25℃下，將1,1-二甲氧基-N,N-二甲基甲胺(58.8mL，442.79mmol)添加至在DMF(46.4mL)中的外消旋-8-溴-1-(3,3-二甲基四氫-2H-哌喃-4-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮(16.66g，44.28mmol)中。將所得的漿液在80℃下攪拌48小時。將反應冷卻至室溫並在真空下過濾沈澱物。將固體用水洗滌並在真空烘箱中乾燥過夜，以得到外消旋-8-溴-1-(3,3-二甲基四氫-2H-哌喃-4-基)-3-甲基-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮(13.24g，77%)。NMR譜： 1H NMR(500MHz,CDCl3)0.89(3H,s),1.32(3H,s),1.73(1H,ddt),3.41(1H,d),3.57(3H,s),3.58-3.64(1H,m),3.68(1H,dd),3.7-3.8(1H,m),4.27(1H,ddd),4.82(1H,dd),7.67(1H,dd),8.03(1H,d),8.55(1H,d),8.71(1H,s)。質譜：m/z(ES+)[M+H]+=390。 At 25 ° C, 1,1-dimethoxy-N, N-dimethylmethylamine (58.8 mL, 442.79 mmol) was added to racemic-8-bromo-1 in DMF (46.4 mL) -(3,3-dimethyltetrahydro-2H-piperan-4-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one (16.66g, 44.28 mmol). The resulting slurry was stirred at 80 ° C for 48 hours. The reaction was cooled to room temperature and the precipitate was filtered under vacuum. The solid was washed with water and dried in a vacuum oven overnight to obtain racemic-8-bromo-1- (3,3-dimethyltetrahydro-2H-piperan-4-yl) -3-methyl- 1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one (13.24g, 77%). NMR spectrum: 1 H NMR (500 MHz, CDCl 3 ) 0.89 (3H, s), 1.32 (3H, s), 1.73 (1H, ddt), 3.41 (1H, d), 3.57 (3H, s), 3.58-3.64 (1H, m), 3.68 (1H, dd), 3.7-3.8 (1H, m), 4.27 (1H, ddd), 4.82 (1H, dd), 7.67 (1H, dd), 8.03 (1H, d), 8.55 (1H, d), 8.71 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 390.

外消旋-8-溴-1-(3,3-二甲基四氫-2H-哌喃-4-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮 Racemic-8-bromo-1- (3,3-dimethyltetrahydro-2H-piperan-4-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinoline Lin-2-one

將三乙胺(18.51mL，132.81mmol)添加至在DMF(100mL)中的外消旋-6-溴-4-((3,3-二甲基四氫-2H-哌喃-4-基)胺基)喹啉-3-甲酸(16.79g，44.27mmol)中。將所得的懸浮液攪拌15分鐘，然後添加二苯基磷醯基疊氮化物(11.45mL，53.13mmol)，並且將懸浮液在室溫下再攪拌30分鐘，然後在60℃下攪拌2小時。將反應冷卻至室溫並用水稀釋。將沈澱物在真空下過濾並在真空烘箱中乾燥，以得到8-溴-1-(3,3-二甲基四氫-2H-哌喃-4-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮(18.42g，111%)。將該物質不經進一步純化而在後續步驟中使用。NMR譜： 1H NMR(500MHz,DMSO-d6)0.73(3H,s),1.13(3H,s),1.69-1.76(1H,m),3.33(1H,s),3.50(2H,qd),3.59-3.68(1H,m),4.04(1H,dd),4.84(1H,dd),7.72(1H,dd),7.94(1H,d),8.62-8.7(2H,m),11.60(1H,s)。質譜：m/z(ES+)[M+H]+=376。 Triethylamine (18.51 mL, 132.81 mmol) was added to racemic-6-bromo-4-((3,3-dimethyltetrahydro-2H-piperan-4-yl) in DMF (100 mL) ) Amino) quinoline-3-carboxylic acid (16.79 g, 44.27 mmol). The resulting suspension was stirred for 15 minutes, then diphenylphosphoryl azide (11.45 mL, 53.13 mmol) was added, and the suspension was stirred at room temperature for another 30 minutes and then at 60 ° C. for 2 hours. The reaction was cooled to room temperature and diluted with water. The precipitate was filtered under vacuum and dried in a vacuum oven to give 8-bromo-1- (3,3-dimethyltetrahydro-2H-piperan-4-yl) -1,3-dihydro- 2H-imidazo [4,5-c] quinolin-2-one (18.42g, 111%). This material was used in subsequent steps without further purification. NMR spectrum: 1 H NMR (500MHz, DMSO-d6) 0.73 (3H, s), 1.13 (3H, s), 1.69-1.76 (1H, m), 3.33 (1H, s), 3.50 (2H, qd), 3.59-3.68 (1H, m), 4.04 (1H, dd), 4.84 (1H, dd), 7.72 (1H, dd), 7.94 (1H, d), 8.62-8.7 (2H, m), 11.60 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 376.

外消旋-6-溴-4-((3,3-二甲基四氫-2H-哌喃-4-基)胺基)喹啉-3-甲酸 Racemic-6-bromo-4-((3,3-dimethyltetrahydro-2H-piperan-4-yl) amino) quinoline-3-carboxylic acid

向在THF(116mL)中的外消旋-6-溴-4-((3,3-二甲基四氫-2H-哌喃-4-基)胺基)喹啉-3-甲酸乙酯(19.66g，48.27mmol) 中添加2M NaOH(97mL，193.07mmol)，並將反應加熱至60℃持續3小時。將該反應冷卻至室溫並在減壓下去除溶劑，以得到棕色溶液。添加2M HCl直至pH 4，並且將所得固體在真空下過濾，並在真空烘箱中乾燥過夜，以得到呈淺黃色固體的外消旋-6-溴-4-((3,3-二甲基四氫-2H-哌喃-4-基)胺基)喹啉-3-甲酸(16.79g，92%)。質譜：m/z(ES+)[M+H]+=379。 To racemic-6-bromo-4-((3,3-dimethyltetrahydro-2H-piperan-4-yl) amino) quinoline-3-carboxylic acid ethyl ester in THF (116 mL) (19.66 g, 48.27 mmol) was added 2M NaOH (97 mL, 193.07 mmol), and the reaction was heated to 60 ° C. for 3 hours. The reaction was cooled to room temperature and the solvent was removed under reduced pressure to obtain a brown solution. 2M HCl was added until pH 4, and the resulting solid was filtered under vacuum and dried in a vacuum oven overnight to give racemic-6-bromo-4-((3,3-dimethyl) as a pale yellow solid Tetrahydro-2H-piperan-4-yl) amino) quinoline-3-carboxylic acid (16.79 g, 92%). Mass spectrum: m / z (ES +) [M + H] + = 379.

外消旋-6-溴-4-((3,3-二甲基四氫-2H-哌喃-4-基)胺基)喹啉-3-甲酸乙酯 Racemic-6-bromo-4-((3,3-dimethyltetrahydro-2H-piperan-4-yl) amino) quinoline-3-carboxylic acid ethyl ester

將三乙胺(26.9mL，193.03mmol)添加到在MeCN(134mL)中的3,3-二甲基四氫-2H-哌喃-4-胺鹽酸鹽(10.39g，62.73mmol)和6-溴-4-氯喹啉-3-甲酸乙酯(15.18g，48.26mmol)中。將該反應混合物在90℃加熱4小時。將該反應冷卻至室溫並將沈澱物在真空下過濾並用水(300mL)洗滌，以得到呈白色固體的外消旋-6-溴-4-((3,3-二甲基四氫-2H-哌喃-4-基)胺基)喹啉-3-甲酸乙酯(11.0g，56%)。將濾液濃縮並用DCM(400mL)萃取。將有機層在相分離器上乾燥並在減壓下濃縮，以得到第二批呈橙色固體的外消旋-6-溴-4-((3,3-二甲基四氫-2H-哌喃-4-基)胺基)喹啉-3-甲酸乙酯(8.0g，41%)。將這兩個批次合併並用於隨後的步驟中。NMR譜： 1H NMR(500MHz,CDCl3)0.85(3H,s),1.21(3H,s),1.44(3H,t),1.95(2H,dd),3.19(1H,d),3.48-3.59(2H,m),3.92 (1H,td),4.04(1H,dt),4.42(2H,q),7.74(1H,dd),7.84(1H,d),8.22(1H,d),8.96(1H,d),9.12(1H,s)。質譜：m/z(ES+)[M+H]+=407.2 Triethylamine (26.9 mL, 193.03 mmol) was added to 3,3-dimethyltetrahydro-2H-piperan-4-amine hydrochloride (10.39 g, 62.73 mmol) and 6 in MeCN (134 mL) -Ethyl bromo-4-chloroquinoline-3-carboxylate (15.18 g, 48.26 mmol). The reaction mixture was heated at 90 ° C for 4 hours. The reaction was cooled to room temperature and the precipitate was filtered under vacuum and washed with water (300 mL) to give racemic-6-bromo-4-((3,3-dimethyltetrahydro- 2H-piperan-4-yl) amino) quinoline-3-carboxylic acid ethyl ester (11.0 g, 56%). The filtrate was concentrated and extracted with DCM (400 mL). The organic layer was dried on a phase separator and concentrated under reduced pressure to obtain a second batch of racemic-6-bromo-4-((3,3-dimethyltetrahydro-2H-piper Furan-4-yl) amino) quinoline-3-carboxylic acid ethyl ester (8.0 g, 41%). The two batches are combined and used in subsequent steps. NMR spectrum: 1 H NMR (500 MHz, CDCl 3 ) 0.85 (3H, s), 1.21 (3H, s), 1.44 (3H, t), 1.95 (2H, dd), 3.19 (1H, d), 3.48-3.59 (2H, m), 3.92 (1H, td), 4.04 (1H, dt), 4.42 (2H, q), 7.74 (1H, dd), 7.84 (1H, d), 8.22 (1H, d), 8.96 ( 1H, d), 9.12 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 407.2

實例12 Example 12

(S)-8-(6-(甲氧基甲基)吡啶-3-基)-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -1- (tetrahydro-2H-piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one

在氮氣下，將三乙胺(1.063mL，7.62mmol)添加到在DMF(10mL)中的(S)-6-(6-(甲氧基甲基)吡啶-3-基)-4-((四氫-2H-哌喃-3-基)胺基)喹啉-3-甲酸(1g，2.54mmol)中。將所得的懸浮液攪拌15分鐘，然後添加二苯基磷醯基疊氮化物(0.657mL，3.05mmol)，並將懸浮液在室溫下再攪拌30分鐘，然後在60℃下攪拌1小時。將該反應混合物倒在冰(約100mL)上，並且濾出固體並乾燥，然後在Et2O中攪拌兩小時。濾出固體並乾燥，以得到呈白色固體的(S)-8-(6-(甲氧基甲基)吡啶-3-基)-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮(0.967g，97%)。NMR譜： 1H NMR(500MHz,DMSO)δ 1.66-1.94(2H,m),2.16(1H,d),2.58-2.7(1H,m),3.35-3.4(1H,m),3.41(3H,s),3.92(1H,d),4.13(1H,dd),4.23(1H,t),4.58(2H,s),4.8-5.06(1H,m),7.49-7.74(1H,m),7.97(1H,dd),8.13(1H,d),8.23(1H,dd),8.40(1H,d),8.66(1H,s),8.97(1H,d),11.57(1H,s)。質譜：m/z(ES+)[M+H]+=391。 Under nitrogen, triethylamine (1.063 mL, 7.62 mmol) was added to ( S ) -6- (6- (methoxymethyl) pyridin-3-yl) -4- () in DMF (10 mL) (Tetrahydro-2H-piperan-3-yl) amino) quinoline-3-carboxylic acid (1 g, 2.54 mmol). The resulting suspension was stirred for 15 minutes, then diphenylphosphoryl azide (0.657 mL, 3.05 mmol) was added, and the suspension was stirred at room temperature for another 30 minutes and then at 60 ° C. for 1 hour. The reaction mixture was poured onto ice (about 100 mL), and the solid was filtered and dried, and then stirred in Et 2 O for two hours. The solid was filtered off and dried to give ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -1- (tetrahydro-2H-piperan-3-yl) as a white solid -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one (0.967g, 97%). NMR spectrum: 1 H NMR (500MHz, DMSO) δ 1.66-1.94 (2H, m), 2.16 (1H, d), 2.58-2.7 (1H, m), 3.35-3.4 (1H, m), 3.41 (3H, s), 3.92 (1H, d), 4.13 (1H, dd), 4.23 (1H, t), 4.58 (2H, s), 4.8-5.06 (1H, m), 7.49-7.74 (1H, m), 7.97 (1H, dd), 8.13 (1H, d), 8.23 (1H, dd), 8.40 (1H, d), 8.66 (1H, s), 8.97 (1H, d), 11.57 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 391.

還可以藉由溶解於少量二氯甲烷中並用等量的溶解於少量二氯甲烷中的甲磺酸處理，去除溶劑並且然後在Et2O中攪拌殘餘物，隨後過濾，將該物質分離為甲磺酸鹽。NMR譜： 1H NMR(500MHz,DMSO-d6)δ 1.75-1.93(2H,m),2.16-2.29(1H,m),2.36(3H,s),2.59-2.76(1H,m),3.31-3.48(4H,m),3.94(1H,d),4.13-4.31(2H,m),4.64(2H,s),5.05-5.15(1H,m),7.71(1H,d),8.22-8.49(3H,m),8.58(1H,s),9.06(2H,d),12.47(1H,s)。質譜：m/z(ES+)[M+H]+=391。 It is also possible to separate the material into methyl alcohol by dissolving in a small amount of dichloromethane and treating with an equivalent amount of methanesulfonic acid dissolved in a small amount of methylene chloride, removing the solvent and then stirring the residue in Et 2 O, followed by filtration. Sulfonate. NMR spectrum: 1 H NMR (500 MHz, DMSO-d6) δ 1.75-1.93 (2H, m), 2.16-2.29 (1H, m), 2.36 (3H, s), 2.59-2.76 (1H, m), 3.31- 3.48 (4H, m), 3.94 (1H, d), 4.13-4.31 (2H, m), 4.64 (2H, s), 5.05-5.15 (1H, m), 7.71 (1H, d), 8.22-8.49 ( 3H, m), 8.58 (1H, s), 9.06 (2H, d), 12.47 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 391.

(S)-6-(6-(甲氧基甲基)吡啶-3-基)-4-((四氫-2H-哌喃-3-基)胺基)喹啉-3-甲酸( S ) -6- (6- (methoxymethyl) pyridin-3-yl) -4-((tetrahydro-2H-piperan-3-yl) amino) quinoline-3-carboxylic acid

將(S)-6-(6-(甲氧基甲基)吡啶-3-基)-4-((四氫-2H-哌喃-3-基)胺基)喹啉-3-甲酸乙酯(1.64g，3.89mmol)在THF(40mL)中的溶液用在水(20mL)中的氫氧化鈉(0.311g，7.78mmol)處理，並且將溶液在60℃下攪拌3小時，然後允許冷卻。將有機溶劑在減壓下去除，並且用2M HCl將水溶液調節至pH 1，給出沈澱物，將該沈澱物濾出，用水和Et2O充分洗滌，然後乾燥，以得到呈白色固體的(S)-6-(6-(甲氧基甲基)吡啶-3-基)-4-((四氫-2H-哌喃-3-基)胺基)喹啉-3-甲酸(1.080g，70.5%)。質譜：m/z(ES+)[M+H]+=394。 ( S ) -6- (6- (methoxymethyl) pyridin-3-yl) -4-((tetrahydro-2H-piperan-3-yl) amino) quinoline-3-carboxylic acid ethyl A solution of the ester (1.64 g, 3.89 mmol) in THF (40 mL) was treated with sodium hydroxide (0.311 g, 7.78 mmol) in water (20 mL), and the solution was stirred at 60 ° C. for 3 hours, and then allowed to cool . The organic solvent was removed under reduced pressure, and the aqueous solution was adjusted to pH 1 with 2M HCl to give a precipitate, which was filtered off, washed thoroughly with water and Et 2 O, and then dried to obtain a white solid ( S ) -6- (6- (methoxymethyl) pyridin-3-yl) -4-((tetrahydro-2H-piperan-3-yl) amino) quinoline-3-carboxylic acid (1.080g , 70.5%). Mass spectrum: m / z (ES +) [M + H] + = 394.

(S)-6-(6-(甲氧基甲基)吡啶-3-基)-4-((四氫-2H-哌喃-3-基)胺( S ) -6- (6- (methoxymethyl) pyridin-3-yl) -4-((tetrahydro-2H-piperan-3-yl) amine 基)喹啉-3-甲酸乙酯Yl) quinoline-3-carboxylic acid ethyl ester

向4-氯-6-(6-(甲氧基甲基)吡啶-3-基)喹啉-3-甲酸乙酯(2g，5.61mmol)和(S)-四氫-2H-哌喃-3-胺鹽酸鹽(0.926g，6.73mmol)在DMF(10mL)中的混合物中添加DIPEA(3.5mL，20.04mmol)，並且將溶液在80℃下攪拌3小時，然後允許冷卻。將該反應混合物與水(100mL)一起攪拌，並且濾出固體，用水充分洗滌並吸乾。將粗產物藉由FCC進行純化，洗提梯度為在DCM中的0至3% 2N甲醇胺，並且將純級分蒸發至乾燥，以得到呈白色固體的(S)-6-(6-(甲氧基甲基)吡啶-3-基)-4-((四氫-2H-哌喃-3-基)胺基)喹啉-3-甲酸乙酯(1.640g，69.4%)。NMR譜： 1H NMR(500MHz,DMSO-d6)δ 1.36(3H,t),1.47-1.64(1H,m),1.67-1.94(2H,m),1.96-2.2(1H,m),3.40(3H,s),3.56(1H,dd),3.61(2H,t),3.87(1H,dd),4.22(1H,dd),4.36(2H,q),4.56(2H,s),7.54(1H,dd),7.95(1H,d),8.10(1H,dd),8.19(1H,dd),8.40(1H,d),8.91(1H,s),8.92-8.93(1H,m),8.94(1H,dd)。質譜：m/z(ES+)[M+H]+=422。 To 4-chloro-6- (6- (methoxymethyl) pyridin-3-yl) quinoline-3-carboxylic acid ethyl ester (2g, 5.61mmol) and ( S ) -tetrahydro-2H-piran- To a mixture of 3-amine hydrochloride (0.926 g, 6.73 mmol) in DMF (10 mL), DIPEA (3.5 mL, 20.04 mmol) was added, and the solution was stirred at 80 ° C. for 3 hours, and then allowed to cool. The reaction mixture was stirred with water (100 mL), and the solid was filtered off, washed thoroughly with water and sucked dry. The crude product was purified by FCC with an elution gradient of 0 to 3% 2N methanolamine in DCM, and the pure fraction was evaporated to dryness to give ( S ) -6- (6- ( Methoxymethyl) pyridin-3-yl) -4-((tetrahydro-2H-piperan-3-yl) amino) quinoline-3-carboxylic acid ethyl ester (1.640 g, 69.4%). NMR spectrum: 1 H NMR (500MHz, DMSO-d6) δ 1.36 (3H, t), 1.47-1.64 (1H, m), 1.67-1.94 (2H, m), 1.96-2.2 (1H, m), 3.40 ( 3H, s), 3.56 (1H, dd), 3.61 (2H, t), 3.87 (1H, dd), 4.22 (1H, dd), 4.36 (2H, q), 4.56 (2H, s), 7.54 (1H , dd), 7.95 (1H, d), 8.10 (1H, dd), 8.19 (1H, dd), 8.40 (1H, d), 8.91 (1H, s), 8.92-8.93 (1H, m), 8.94 ( 1H, dd). Mass spectrum: m / z (ES +) [M + H] + = 422.

4-氯-6-(6-(甲氧基甲基)吡啶-3-基)喹啉-3-甲酸乙酯4-chloro-6- (6- (methoxymethyl) pyridin-3-yl) quinoline-3-carboxylic acid ethyl ester

在氮氣下，將四(三苯基膦)鈀(0)(2.204g，1.91mmol)添加到在1,4-二(100mL)和水(20.00mL)中的6- 溴-4-氯喹啉-3-甲酸乙酯(6.0g，19.07mmol)、2-(甲氧基甲基)-5-(4,4,5,5-四甲基-1,3,2-二氧雜環戊硼烷-2-基)吡啶(10.30g，24.80mmol)以及碳酸銫(12.43g，38.15mmol)中。將所得的懸浮液在120℃下攪拌3小時。將粗產物藉由FCC進行純化，洗提梯度為在庚烷中的25%至100% EtOAc。將純級分蒸發至乾燥，以得到呈奶油色固體的4-氯-6-(6-(甲氧基甲基)吡啶-3-基)喹啉-3-甲酸乙酯(3.10g，45.6%)。NMR譜： 1H NMR(400MHz,CDCl3)δ 1.48(3H,t),3.54(3H,s),4.52(2H,q),4.68(2H,s),7.59(1H,d),8.02-8.09(2H,m),8.26(1H,d),8.58(1H,d),8.94(1H,d),9.22(1H,s)。質譜：m/z(ES+)[M+H]+=357。 Under nitrogen, tetrakis (triphenylphosphine) palladium (0) (2.204g, 1.91mmol) was added to 1,4-bis (100 mL) and 6-bromo-4-chloroquinoline-3-carboxylic acid ethyl ester (6.0 g, 19.07 mmol), 2- (methoxymethyl) -5- (4,4, 5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) pyridine (10.30 g, 24.80 mmol) and cesium carbonate (12.43 g, 38.15 mmol). The resulting suspension was stirred at 120 ° C for 3 hours. The crude product was purified by FCC with an elution gradient of 25% to 100% EtOAc in heptane. The pure fractions were evaporated to dryness to give 4-chloro-6- (6- (methoxymethyl) pyridin-3-yl) quinoline-3-carboxylic acid ethyl ester (3.10 g, 45.6) as a cream solid %). NMR spectrum: 1 H NMR (400MHz, CDCl 3 ) δ 1.48 (3H, t), 3.54 (3H, s), 4.52 (2H, q), 4.68 (2H, s), 7.59 (1H, d), 8.02- 8.09 (2H, m), 8.26 (1H, d), 8.58 (1H, d), 8.94 (1H, d), 9.22 (1H, s). Mass spectrum: m / z (ES +) [M + H] + = 357.

1-(3,3-二甲基四氫-2H-哌喃-4-基)-8-(6-(甲氧基甲基)吡啶-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮1- (3,3-Dimethyltetrahydro-2H-piperan-4-yl) -8- (6- (methoxymethyl) pyridin-3-yl) -1H-imidazo [4,5 -c] quinoline-2 (3H) -one

在0℃下，將三氯異氰尿酸(0.567g，2.44mmol)分批添加到4-((3,3-二甲基四氫-2H-哌喃-4-基)胺基)-6-(6-(甲氧基甲基)吡啶-3-基)喹啉-3-甲醯胺(2.05g，4.88mmol)和1,8-二氮雜二環[5.4.0]十一-7-烯(1.458mL，9.75mmol)在MeOH(2mL)中的攪拌懸浮液中。將所得的懸浮液攪拌至室溫持續4小時。將該反應混合物蒸發至乾燥並再溶於DCM(75mL)中，並用水(50mL)洗滌。將有機層用相分離柱乾燥並蒸發，以得到粗產物。將該物質不經進一步純化進行下一步驟。質譜：m/z(ES+)[M+H]+=419。 At 0 ° C, trichloroisocyanuric acid (0.567 g, 2.44 mmol) was added portionwise to 4-((3,3-dimethyltetrahydro-2H-piperan-4-yl) amino) -6 -(6- (methoxymethyl) pyridin-3-yl) quinoline-3-carboxamide (2.05g, 4.88mmol) and 1,8-diazabicyclo [5.4.0] undec- 7-ene (1.458 mL, 9.75 mmol) in a stirred suspension in MeOH (2 mL). The resulting suspension was stirred to room temperature for 4 hours. The reaction mixture was evaporated to dryness and redissolved in DCM (75 mL), and washed with water (50 mL). The organic layer was dried with a phase separation column and evaporated to obtain a crude product. This material was carried on to the next step without further purification. Mass spectrum: m / z (ES +) [M + H] + = 419.

4-((3,3-二甲基四氫-2H-哌喃-4-基)胺基)-6-(6-(甲氧基甲基)吡啶-3-基)喹啉-3-甲醯胺4-((3,3-dimethyltetrahydro-2H-piperan-4-yl) amino) -6- (6- (methoxymethyl) pyridin-3-yl) quinoline-3- Formamide

將3,3-二甲基四氫-2H-哌喃-4-胺(0.694g，5.37mmol)緩慢添加到在DMF(11mL)中的4-氯-6-(6-(甲氧基甲基)吡啶-3-基)喹啉-3-甲醯胺(2g，4.88mmol)和三乙胺(0.408mL，2.93mmol)中。將所得的漿液在90℃下攪拌4小時。添加另外的300mg的3,3-二甲基四氫-2H-哌喃-4-胺並再攪拌16小時過夜。在減壓下去除DMF並將棕色油狀物用DCM(50mL)稀釋。將有機物用水(50mL)洗滌並經相分離器乾燥。在減壓下去除溶劑，以得到棕色固體。物質不經進一步純化即可進行。質譜：m/z(ES+)[M+H]+=421。 3,3-Dimethyltetrahydro-2H-piperan-4-amine (0.694 g, 5.37 mmol) was slowly added to 4-chloro-6- (6- (methoxymethoxy) in DMF (11 mL) Yl) pyridin-3-yl) quinoline-3-carboxamide (2g, 4.88mmol) and triethylamine (0.408mL, 2.93mmol). The resulting slurry was stirred at 90 ° C for 4 hours. An additional 300 mg of 3,3-dimethyltetrahydro-2H-piperan-4-amine was added and stirred for another 16 hours overnight. DMF was removed under reduced pressure and the brown oil was diluted with DCM (50 mL). The organics were washed with water (50 mL) and dried through a phase separator. The solvent was removed under reduced pressure to obtain a brown solid. The material can be carried on without further purification. Mass spectrum: m / z (ES +) [M + H] + = 421.

4-氯-6-(6-(甲氧基甲基)吡啶-3-基)喹啉-3-甲醯胺4-chloro-6- (6- (methoxymethyl) pyridin-3-yl) quinoline-3-carboxamide

將4-氯-6-(6-(甲氧基甲基)吡啶-3-基)喹啉-3-碳醯氯(15.62g，45mmol)在MeCN(200mL)中的漿液分批添加至氫氧化銨(58.4mL，450.00mmol)中，保持內部溫度在0℃下並攪拌1小時。過濾所沈澱的固體，用水洗滌並乾燥，以得到呈米色固體的粗的4-氯-6-(6-(甲氧基甲基)吡啶-3-基)喹啉-3-甲醯胺(10.90g，73.9%)。質譜：m/z(ES+)[M+H]+=328。 A slurry of 4-chloro-6- (6- (methoxymethyl) pyridin-3-yl) quinoline-3-carbamide chloride (15.62 g, 45 mmol) in MeCN (200 mL) was added portionwise to hydrogen In ammonium oxide (58.4 mL, 450.00 mmol), the internal temperature was kept at 0 ° C and stirred for 1 hour. The precipitated solid was filtered, washed with water and dried to obtain crude 4-chloro-6- (6- (methoxymethyl) pyridin-3-yl) quinoline-3-carboxamide as a beige solid ( 10.90g, 73.9%). Mass spectrum: m / z (ES +) [M + H] + = 328.

4-氯-6-(6-(甲氧基甲基)吡啶-3-基)喹啉-3-碳醯氯4-chloro-6- (6- (methoxymethyl) pyridin-3-yl) quinoline-3-carbamide

在室溫下，將DMF(0.352mL，4.54mmol)添加到6-(6-(甲氧基甲基)吡啶-3-基)-4-側氧基-1,4-二氫喹啉-3-甲酸(14.1g，45.44mmol)和亞硫醯氯(66.3mL，908.79mmol)中。將所得的溶液在75℃下攪拌持續小時，伴隨著氣體藉由水洗滌器。將所得的混合物蒸發至乾燥並與甲苯(2 x 100mL)共沸，以得到粗的呈棕色固體的4-氯-6-(6-(甲氧基甲基)吡啶-3-基)喹啉-3-碳醯氯，將其不經進一步純化用於下一反應中。 At room temperature, DMF (0.352 mL, 4.54 mmol) was added to 6- (6- (methoxymethyl) pyridin-3-yl) -4-oxo-1,4-dihydroquinoline- 3-Formic acid (14.1 g, 45.44 mmol) and sulfenyl chloride (66.3 mL, 908.79 mmol). The resulting solution was stirred at 75 ° C for an hour, accompanied by gas through a water scrubber. The resulting mixture was evaporated to dryness and azeotroped with toluene (2 x 100 mL) to give crude 4-chloro-6- (6- (methoxymethyl) pyridin-3-yl) quinoline as a brown solid -3-Carboacetyl chloride, which was used in the next reaction without further purification.

6-(6-(甲氧基甲基)吡啶-3-基)-4-側氧基-1,4-二氫喹啉-3-甲酸6- (6- (methoxymethyl) pyridin-3-yl) -4-oxo-1,4-dihydroquinoline-3-carboxylic acid

將氫氧化鈉(40.0mL，80.00mmol)添加到6-(6-(甲氧基甲基)吡啶-3-基)-4-側氧基-1,4-二氫喹啉-3-甲酸乙酯(5.41g，16mmol)在乙醇(50mL)中的攪拌懸浮液中。將所得的懸浮液在100℃下攪拌30分鐘。添加水(50mL)，然後添加HCl(2N)以獲得pH 3。過濾沈澱物，用水洗滌並在真空下乾燥，以得到呈灰色固體的6-(6-(甲氧基甲基)吡啶-3-基)-4-側氧基-1,4-二氫喹啉-3-甲酸(4.15g，84%)。質譜：m/z(ES+)[M+H]+=311。 Sodium hydroxide (40.0 mL, 80.00 mmol) was added to 6- (6- (methoxymethyl) pyridin-3-yl) -4-oxo-1,4-dihydroquinoline-3-carboxylic acid Ethyl ester (5.41 g, 16 mmol) in a stirred suspension in ethanol (50 mL). The resulting suspension was stirred at 100 ° C for 30 minutes. Water (50 mL) was added, followed by HCl (2N) to obtain pH 3. The precipitate was filtered, washed with water and dried under vacuum to give 6- (6- (methoxymethyl) pyridin-3-yl) -4-oxo-1,4-dihydroquino as a gray solid Porphyrin-3-carboxylic acid (4.15g, 84%). Mass spectrum: m / z (ES +) [M + H] + = 311.

6-(6-(甲氧基甲基)吡啶-3-基)-4-側氧基-1,4-二氫喹啉-3-甲酸乙酯6- (6- (methoxymethyl) pyridin-3-yl) -4-oxo-1,4-dihydroquinoline-3-carboxylic acid ethyl ester

在氮氣下，將四氯鈀酸鈉(II)(2.68g，9.12mmol)和3-(二-三級丁基膦基)丙烷-1-磺酸(4.89g，18.24mmol)一次性添加至在充分脫氣的二(500mL)和水(125mL)中的6-溴-4-側氧基-1,4-二氫喹啉-3-甲酸乙酯(54g，182.36mmol)、2-(甲氧基甲基)-5-(4,4,5,5-四甲基-1,3,2-二氧雜環戊硼烷-2-基)吡啶(83g，200.60mmol)以及K2CO3(76g，547.08mmol)中。將所得的混合物在85℃下攪拌2小時，然後冷卻至室溫。過濾沈澱物並用水洗滌，以給出粗的呈黃色固體的6-(6-(甲氧基甲基)吡啶-3-基)-4-側氧基-1,4-二氫喹啉-3-甲酸乙酯(38.1g，61.7%)。靜置時，濾液沈澱出更多量的固體物質，將其過濾，以給出呈白色固體的6-(6-(甲氧基甲基)吡啶-3-基)-4-側氧基-1,4-二氫喹啉-3-甲酸乙酯(12g，19.45%)。這兩個樣品都不經進一步純化而使用。質譜：m/z(ES+)[M+H]+=339。 Under nitrogen, sodium tetrachloropalladium (II) (2.68 g, 9.12 mmol) and 3- (di-tertiary butylphosphino) propane-1-sulfonic acid (4.89 g, 18.24 mmol) were added at once to In the fully degassed two (500mL) and 6-bromo-4-oxo-1,4-dihydroquinoline-3-carboxylic acid ethyl ester (54g, 182.36mmol) and 2- (methoxymethyl) in water (125mL) -5- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) pyridine (83g, 200.60mmol) and K 2 CO 3 (76g, 547.08 mmol). The resulting mixture was stirred at 85 ° C for 2 hours, and then cooled to room temperature. The precipitate was filtered and washed with water to give crude 6- (6- (methoxymethyl) pyridin-3-yl) -4-oxo-1,4-dihydroquinoline- as a yellow solid Ethyl 3-formate (38.1g, 61.7%). When standing, the filtrate precipitated a greater amount of solid material, which was filtered to give 6- (6- (methoxymethyl) pyridin-3-yl) -4-oxo- as a white solid 1,4-Dihydroquinoline-3-carboxylic acid ethyl ester (12g, 19.45%). Both samples were used without further purification. Mass spectrum: m / z (ES +) [M + H] + = 339.

6-溴-4-側氧基-1H-喹啉-3-甲酸乙酯6-Bromo-4-oxo-1H-quinoline-3-carboxylic acid ethyl ester

將二苯醚(870mL)加熱至240℃，然後分批添加{[(4-溴苯基)胺基]亞甲基}丙二酸二乙酯(75g，219.18mmol)。將該混合物在裝有迪安-斯達克(dean-stark)裝置的燒瓶中在240℃下攪拌60分鐘。冷卻後，將該混合物用Et2O稀釋，並且將固體藉由過濾收集，用Et2O洗滌並乾燥，以得到呈米色結晶固體的6-溴-4-側 氧基-1H-喹啉-3-甲酸乙酯(59.9g)，將其不進行純化或表徵而使用。 Diphenyl ether (870 mL) was heated to 240 ° C., and then {[((4-bromophenyl) amino] methylene} malonic acid diethyl ester (75 g, 219.18 mmol) was added in portions. The mixture was stirred at 240 ° C for 60 minutes in a flask equipped with a dean-stark device. After cooling, the mixture was diluted with Et 2 O, and the solid was collected by filtration, washed with Et 2 O and dried to obtain 6-bromo-4-oxo-1H-quinoline- as a beige crystalline solid 3-Ethyl formate (59.9 g), which was used without purification or characterization.

{[(4-溴苯基)胺基]亞甲基}丙二酸二乙酯{[(4-Bromophenyl) amino] methylene} diethyl malonate

將2-(乙氧基亞甲基)丙二酸二乙酯(71.5mL，354mmol)添加至在EtOH(420mL)中的4-溴苯胺(42g，244mmol)中，並且將所得的混合物在80℃下攪拌過夜。在冷卻至10℃之後，將白色固體藉由過濾收集，用庚烷洗滌並且乾燥，以得到呈白色結晶固體的{[(4-溴苯基)胺基]亞甲基}丙二酸二乙酯(75g，90%)。 1 H NMR譜(500MHz,DMSO-d6)δ 1.25(6H,s),4.10-4.27(4H,m),7.38(2H,d),7.57(2H,d),8.37(1H,br s)。質譜：m/z(ES+)[M+H]+=342,344。 Diethyl 2- (ethoxymethylene) malonate (71.5 mL, 354 mmol) was added to 4-bromoaniline (42 g, 244 mmol) in EtOH (420 mL), and the resulting mixture was added at 80 Stir overnight at ℃. After cooling to 10 ° C, the white solid was collected by filtration, washed with heptane and dried to give {[(4-bromophenyl) amino] methylene} malonic acid diethyl as a white crystalline solid Ester (75g, 90%). 1 H NMR spectrum (500 MHz, DMSO-d6) δ 1.25 (6H, s), 4.10-4.27 (4H, m), 7.38 (2H, d), 7.57 (2H, d), 8.37 (1H, br s). Mass spectrum: m / z (ES +) [M + H] + = 342,344.

生物學測定 Biological assay

以下測定用於測量本發明的該等化合物的效果：a)ATM細胞效價測定；b)PI3K細胞效價測定；c)mTOR細胞效價測定；d)ATR細胞效價測定；e)：DNAPK細胞效價測定。在測定的描述中，通常： The following assays are used to measure the effect of the compounds of the invention: a) ATM cell titer assay; b) PI3K cell titer assay; c) mTOR cell titer assay; d) ATR cell titer assay; e): DNAPK Cell titer determination. In the description of the determination, usually:

i.使用以下縮寫：4NQO=4-硝基喹啉N-氧化物；Ab=抗體；BSA=牛血清白蛋白；CO2=二氧化碳；DMEM=杜氏改良的伊格爾氏培養基(Dulbecco's Modified Eagle Medium)；DMSO= 二甲基亞碸；EDTA=乙二胺四乙酸；EGTA=乙二醇四乙酸；ELISA=酶聯免疫吸附測定；EMEM=伊格爾氏極限必需培養基(Eagle's Minimal Essential Medium)；FBS=胎牛血清；h=小時；HRP=辣根過氧化物酶；i.p.=腹膜內的；PBS=磷酸鹽緩衝鹽水；PBST=磷酸鹽緩衝鹽水/吐溫；TRIS=三(羥基甲基)胺基甲烷；MTS試劑：[3-(4,5-二甲基噻唑-2-基)-5-(3-羧基甲氧基苯基)-2-(4-磺基苯基)-2H-四氮唑，內鹽，以及電子偶聯劑(啡硫酸甲酯)PMS；s.c.皮下。 i. Use the following abbreviations: 4NQO = 4-nitroquinoline N -oxide; Ab = antibody; BSA = bovine serum albumin; CO 2 = carbon dioxide; DMEM = Dulbecco's Modified Eagle Medium ); DMSO = dimethyl sulfoxide; EDTA = ethylenediaminetetraacetic acid; EGTA = ethylene glycol tetraacetic acid; ELISA = enzyme-linked immunosorbent assay; EMEM = Eagle's Minimal Essential Medium; FBS = fetal bovine serum; h = hour; HRP = horseradish peroxidase; ip = intraperitoneal; PBS = phosphate buffered saline; PBST = phosphate buffered saline / Tween; TRIS = tris (hydroxymethyl) Aminomethane; MTS reagent: [3- (4,5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2- (4-sulfophenyl) -2H -Tetrazole, internal salt, and electron coupling agent (coffee Methyl sulfate) PMS; sc subcutaneously.

ii.使用Genedata智慧擬合模型計算IC50值。該IC50值係抑制50%生物學活性的測試化合物的濃度。 II. IC 50 values are calculated using Genedata wisdom fit model. The IC 50 values based inhibitory concentration of the test compound in 50% of the biological activity.

測定a)：ATM細胞效價 Measurement a): ATM cell titer

基本原理：Fundamental:

細胞輻照誘導DNA雙股斷裂和絲胺酸1981的快速分子間自磷酸化，這導致二聚體解離並引發細胞ATM激酶活性。在輻照劑量低至0.5Gy後，細胞中的大多數ATM分子在此位點上快速磷酸化，並且磷酸特異性抗體的結合係在細胞中引入只有少數DNA雙股斷裂後可檢測的。 Cellular irradiation induces DNA double-strand breaks and rapid intermolecular autophosphorylation of serine 1981, which results in dissociation of the dimer and triggers cellular ATM kinase activity. After the irradiation dose is as low as 0.5Gy, most ATM molecules in the cell are rapidly phosphorylated at this site, and the binding system of the phosphate-specific antibody is detectable after introducing a few DNA double-strand breaks into the cell.

pATM測定的基本原理係識別細胞中ATM的抑制劑。先於X射線輻照，將HT29細胞用測試化合物培養1小時。1小時後，將該等細胞固定並用pATM(Ser1981)染色。在測定掃描成像平臺上讀取螢光。 The basic principle of pATM measurement is to identify inhibitors of ATM in cells. Prior to X-ray irradiation, HT29 cells were incubated with the test compound for 1 hour. After 1 hour, the cells were fixed and stained with pATM (Ser1981). Read the fluorescence on the measurement scanning imaging platform.

方法細節：Method details:

將HT29細胞(ECACC #85061109)以每孔3500個細胞的密度接種於384孔測定板(科斯塔#3712)中包含1% L-穀胺醯胺和10% FBS的40μl EMEM培養基中且允許其粘附過夜。次日早晨藉由聲學分配將於100% DMSO中的具有化學式(I)之化合物添加到測定板中。在37℃下和5% CO2下培養1小時後，使用相當於約600cGy的X-RAD 320儀器(PXi)對該等板(多至一次6個)進行輻照。將板返回培養箱中再培養1小時。然後將細胞藉由添加在PBS溶液中的20μl的3.7%甲醛並且在室溫下培養20分鐘來進行固定，之後使用伯騰(Biotek)EL405洗板機，每孔用50μl PBS進行洗滌。然後添加在PBS中的20μl的0.1% Triton X100，並且在室溫下培養20分鐘以透化細胞。接著使用伯騰EL405洗板機，每孔用50μl PBS洗滌該等板一次。 HT29 cells (ECACC # 85061109) were seeded at a density of 3500 cells per well in a 384-well assay plate (Costa # 3712) in 40 μl EMEM medium containing 1% L-glutamine and 10% FBS and allowed to Stick overnight. The next morning the compound of formula (I) in 100% DMSO was added to the assay plate by acoustic distribution. After incubation at 37 ° C. and 5% CO 2 for 1 hour, the plates (up to 6 at a time) were irradiated using an X-RAD 320 instrument (PXi) equivalent to approximately 600 cGy. Return the plate to the incubator for another hour. The cells were then fixed by adding 20 μl of 3.7% formaldehyde in PBS solution and incubated at room temperature for 20 minutes, after which a Biotek EL405 plate washer was used and washed with 50 μl PBS per well. Then 20 μl of 0.1% Triton X100 in PBS was added and incubated at room temperature for 20 minutes to permeabilize the cells. Next, using a Boten EL405 plate washer, the plates were washed once with 50 μl PBS per well.

將磷酸-ATM Ser1981抗體(密理博(Millipore)#MAB3806)稀釋10000倍於包含0.05%聚山梨醇酯/吐溫和3% BSA的PBS中，並且將20μl添加至每個孔，並且在室溫下培養過夜。次日早晨使用伯騰EL405洗板機，每孔用50μl PBS洗滌板三次，並且然後添加20μl在PBS中的二級抗體溶液，該二級抗體溶液包含500倍稀釋的Alexa Fluor® 488山羊抗兔IgG(生命技術公司(Life Technologies)，A11001)以及0.002mg/ml Hoeschst染料(生命技術公司#H-3570)，該PBS包含0.05%聚山梨醇酯/吐溫和3% BSA。在室溫下培養1小時之後，使用伯騰EL405洗板機，每孔用50μl PBS洗滌板三次，並且將板密封並在4℃保持在PBS中直至讀取。使用ArrayScan VTI儀器，使用具有10×物鏡的XF53濾光器來讀取板。使用雙雷射設置來分析細胞核的Hoeschst(405nM)染色和二級抗 體的pSer1981(488nM)染色。 Phospho-ATM Ser1981 antibody (Millipore # MAB3806) was diluted 10,000 times in PBS containing 0.05% polysorbate / Tween and 3% BSA, and 20 μl was added to each well, and at room temperature Incubate overnight. The next morning, using the Boten EL405 plate washer, the plate was washed three times with 50 μl PBS per well, and then 20 μl of a secondary antibody solution in PBS containing 500-fold diluted Alexa Fluor® 488 goat anti-rabbit was added IgG (Life Technologies, A11001) and 0.002 mg / ml Hoeschst dye (Life Technologies # H-3570), the PBS contains 0.05% polysorbate / Tween and 3% BSA. After incubating at room temperature for 1 hour, the plate was washed with a Boten EL405 plate washer, and the plate was washed three times with 50 μl PBS per well, and the plate was sealed and kept in PBS at 4 ° C until reading. Using an ArrayScan VTI instrument, an XF53 filter with a 10 × objective lens was used to read the plate. A dual laser setup was used to analyze Hoeschst (405 nM) staining of nuclei and pSer1981 (488 nM) staining of secondary antibodies.

測定b)：ATR細胞效價 Measurement b): ATR cell titer

基本原理：Fundamental:

ATR係PI 3-激酶-相關的激酶，其響應於DNA損傷或複製阻滯磷酸化絲胺酸或蘇胺酸殘基上的多個底物。Chk1(ATR的下游蛋白激酶)在DNA損傷檢查點控制中起重要作用。Chk1的啟動涉及Ser317和Ser345的磷酸化(後者視為藉由ATR磷酸化/啟動的優先目標)。這係基於細胞的分析，用於藉由在用具有化學式(I)之化合物和UV模擬劑4NQO(西格瑪#N8141)處理之後測量HT29細胞中Chk1(Ser 345)的磷酸化減少，來測量ATR激酶的抑制。 ATR is a PI 3-kinase-related kinase that blocks multiple substrates on phosphorylated serine or threonine residues in response to DNA damage or replication. Chk1 (downstream protein kinase of ATR) plays an important role in DNA damage checkpoint control. The activation of Chk1 involves phosphorylation of Ser317 and Ser345 (the latter is regarded as a priority target for phosphorylation / activation by ATR). This is a cell-based analysis for measuring ATR kinase by measuring the phosphorylation reduction of Chk1 (Ser 345) in HT29 cells after treatment with a compound of formula (I) and a UV mimic 4NQO (Sigma # N8141) Of inhibition.

方法細節：Method details:

將HT29細胞(ECACC #85061109)以每孔6000個細胞的密度接種於384孔測定板(科斯塔#3712)中包含1% L-穀胺醯胺和10% FBS的40μl EMEM培養基中且允許其粘附過夜。次日早晨藉由聲學分配將於100% DMSO中的具有化學式(I)之化合物添加到測定板中。在37℃下和5% CO2下培養1小時後，藉由聲學分配將在100% DMSO中的40nl的3mM 4NQO添加至全部孔中，除了未用4NQO處理以產生空反應對照的最小對照孔。將板返回培養箱中再培養1小時。然後將細胞藉由添加20μl的在PBS溶液中的3.7%甲醛並在室溫下培養20分鐘來固定。然後添加在PBS中的20μl的0.1% Triton X100，並且在室溫下培養10分鐘以透化細胞。接著使 用伯騰EL405洗板機，每孔用50μl PBS洗滌該等板一次。 HT29 cells (ECACC # 85061109) were seeded at a density of 6000 cells per well in a 384-well assay plate (Costa # 3712) in 40 μl EMEM medium containing 1% L-glutamine and 10% FBS and allowed to Stick overnight. The next morning the compound of formula (I) in 100% DMSO was added to the assay plate by acoustic distribution. After incubation at 37 ° C and 5% CO 2 for 1 hour, 40nl of 3mM 4NQO in 100% DMSO was added to all wells by acoustic partitioning, except for the smallest control wells that were not treated with 4NQO to generate empty control . Return the plate to the incubator for another hour. The cells were then fixed by adding 20 μl of 3.7% formaldehyde in PBS solution and incubating at room temperature for 20 minutes. Then 20 μl of 0.1% Triton X100 in PBS was added and incubated at room temperature for 10 minutes to permeabilize the cells. Next, using a Boten EL405 plate washer, the plates were washed once with 50 μl PBS per well.

將磷酸-Chk1 Ser 345抗體(細胞信號傳導技術公司(Cell Signalling Technology)#2348)稀釋150倍於包含0.05%聚山梨醇酯/吐溫的PBS中，並且將15μl添加至每個孔並且在室溫下培養過夜。次日早晨使用伯騰EL405洗板機，每孔用50μl PBS洗滌板三次，且接著添加20μl於PBST中的二級抗體溶液(包含500倍稀釋的Alexa Fluor 488山羊抗兔IgG(分子探針(Molecular Probes)#A-11008)和0.002mg/ml Hoeschst染料(分子探針#H-3570))。在室溫下培養2小時之後，使用伯騰EL405洗板機，每孔用50μl PBS洗滌板三次，並且然後將板用黑色板密封物密封直至讀取。使用ArrayScan VTI儀器，使用具有10×物鏡的XF53濾光器來讀取板。使用雙雷射設置來分析細胞核的Hoeschst(405nm)染色和二級抗體的pChk1(488nm)染色。 Phospho-Chk1 Ser 345 antibody (Cell Signalling Technology # 2348) was diluted 150-fold in PBS containing 0.05% polysorbate / Tween, and 15 μl was added to each well and in the chamber Incubate overnight at temperature. The next morning, using the Boten EL405 plate washer, the plate was washed three times with 50 μl PBS per well, and then 20 μl of a secondary antibody solution (containing 500-fold diluted Alexa Fluor 488 goat anti-rabbit IgG (molecular probe ( Molecular Probes) # A-11008) and 0.002 mg / ml Hoeschst dye (Molecular Probes # H-3570)). After incubation at room temperature for 2 hours, the plate was washed with a Boten EL405 plate washer, 50 μl PBS per well three times, and then the plate was sealed with a black plate sealer until read. Using an ArrayScan VTI instrument, an XF53 filter with a 10 × objective lens was used to read the plate. A dual laser setup was used to analyze Hoeschst (405 nm) staining of nuclei and pChk1 (488 nm) staining of secondary antibodies.

測定c)：PI3K細胞效價 Measurement c): PI3K cell titer

基本原理：Fundamental:

這一測定用於測量細胞中的PI3K-α抑制。PDK1被鑒定為蛋白激酶B(Akt1)的上游啟動環激酶，其對PKB的啟動係必需的。脂質激酶磷酸肌醇3激酶(PI3K)的啟動對於藉由PDK1啟動PKB係至關重要的。 This assay is used to measure PI3K-α inhibition in cells. PDK1 was identified as an upstream promoter loop kinase of protein kinase B (Akt1), which is required for the promoter system of PKB. The activation of the lipid kinase phosphoinositide 3 kinase (PI3K) is essential for the activation of the PKB line by PDK1.

受體酪胺酸激酶配位基刺激後，PI3K被啟動，它將PIP2轉換為PIP3，PIP3由PDK1的PH結構域結合，這導致PDK1向細胞膜募集，在此處在啟動環中的Thr308處磷酸化AKT。 After stimulation by the receptor tyrosine kinase ligand, PI3K is activated, which converts PIP2 to PIP3. PIP3 is bound by the PH domain of PDK1, which causes PDK1 to recruit to the cell membrane, where it is phosphorylated at Thr308 in the activation loop AKT.

這種基於細胞的作用方式測定的目的是識別抑制PDK活性或藉由抑制PI3K活性而導致PDK1向細胞膜募集的化合物。用化合物處理2小時後BT474c細胞中磷酸-Akt(T308)的磷酸化係PDK1的直接度量並且係PI3K活性的間接度量。 The purpose of this cell-based assay is to identify compounds that inhibit PDK activity or lead to the recruitment of PDK1 to the cell membrane by inhibiting PI3K activity. Phospho-Akt (T308) phosphorylation in BT474c cells is a direct measure of PDK1 and an indirect measure of PI3K activity after 2 hours of treatment with the compound.

方法細節：Method details:

將BT474細胞(人類乳腺管癌，ATCC HTB-20)以每孔5600個細胞的密度接種於黑色384孔板(科斯塔，#3712)中包含10% FBS和1%穀胺醯胺的DMEM中並且允許其粘附過夜。 BT474 cells (human breast tube carcinoma, ATCC HTB-20) were seeded at a density of 5600 cells per well in DMEM containing 10% FBS and 1% glutamine in a black 384-well plate (Costa, # 3712) And allow it to stick overnight.

次日早晨藉由聲學分配將於100% DMSO中的化合物的添加到測定板中。在37℃和5% CO2下培養2小時之後，抽吸培養基並且用包含25mM Tris、3mM EDTA、3mM EGTA、50mM氟化鈉、2mM原釩酸鈉、0.27M蔗糖、10mM β-甘油磷酸鹽、5mM焦磷酸鈉、0.5% Triton X-100以及康普利特(complete)蛋白酶抑制劑混合片劑(羅氏(Roche)#04 693 116 001，每50ml溶解緩衝液使用1片)的緩衝液溶解該等細胞。 The next morning the compound in 100% DMSO will be added to the assay plate by acoustic distribution. After incubation at 37 ° C and 5% CO 2 for 2 hours, the medium was aspirated and contained with 25mM Tris, 3mM EDTA, 3mM EGTA, 50mM sodium fluoride, 2mM sodium orthovanadate, 0.27M sucrose, 10mM β-glycerol phosphate , 5mM sodium pyrophosphate, 0.5% Triton X-100, and a complete protease inhibitor mixed tablet (Roche # 04 693 116 001, 1 tablet per 50ml dissolution buffer) Such cells.

20分鐘後，將細胞溶解物轉移到已預塗布於PBS緩衝液中的抗所有AKT抗體的ELISA板(葛萊娜(Greiner)#781077)中，並且用於包含0.05%吐溫20的PBS中的1% BSA阻斷非特異性結合。在4℃下將板培養過夜。次日用包含0.05%吐溫20的PBS緩衝液洗滌該等板並且再與小鼠單克隆抗磷酸AKT T308一起培養2小時。再次如上洗滌板，隨後添加馬抗小鼠HRP結合的二級抗體。在室溫下培養2小時後，洗滌板並且向每一孔中添加QuantaBlu底物工作溶液(賽默科技公司(Thermo Scientific)#15169，根據供 應商說明書製備)。60分鐘後藉由向孔中添加停止溶液使已發展的螢光產物停止。使用帝肯(Tecan)Safire讀板儀分別使用325nm激發波長和420nm發射波長讀取板。除非有所說明，否則在這一ELISA測定中使用來自細胞信號傳導公司(Cell Signalling)(#7144)的Path Scan磷酸AKT(Thr308)夾心ELISA套組中所含的試劑。 After 20 minutes, the cell lysate was transferred to an ELISA plate (Greiner # 781077) that had been pre-coated in PBS buffer against all AKT antibodies, and used in PBS containing 0.05% Tween 20 Of 1% BSA blocks non-specific binding. The plates were incubated overnight at 4 ° C. The following day the plates were washed with PBS buffer containing 0.05% Tween 20 and incubated with mouse monoclonal anti-phosphate AKT T308 for another 2 hours. The plate was washed again as above, followed by the addition of horse anti-mouse HRP-conjugated secondary antibody. After incubating at room temperature for 2 hours, the plate was washed and QuantaBlu substrate working solution (Thermo Scientific # 15169, prepared according to the supplier's instructions) was added to each well. After 60 minutes, the developed fluorescent product was stopped by adding a stop solution to the well. A Tecan Safire plate reader was used to read the plate using an excitation wavelength of 325 nm and an emission wavelength of 420 nm, respectively. Unless stated otherwise, the reagents included in the Path Scan Phosphate AKT (Thr308) sandwich ELISA kit from Cell Signalling (# 7144) were used in this ELISA assay.

測定d)：mTOR細胞效價 Measurement d): mTOR cell titer

基本原理：Fundamental:

在MDA-MB-468細胞系(PTEN無效乳癌人細胞系)中進行磷酸-AKTser473細胞測定。作為缺乏PTEN的結果，pAKT被組成型活化，這消除了誘導磷酸化的刺激的需要。 Phospho-AKTser473 cell assay was performed in MDA-MB-468 cell line (PTEN null breast cancer human cell line). As a result of the lack of PTEN, pAKT is constitutively activated, which eliminates the need for stimulation that induces phosphorylation.

方法細節：Method details:

將MDA-MB-468細胞在細胞培養基中培養，該細胞培養基由DMEM(杜氏改良的伊格爾氏培養基#D6546)、10%(v/v)胎牛血清和1%(v/v)L-穀胺醯胺組成。收穫之後，將細胞分配至黑色384-孔Costar板(#3712，康寧公司(Corning))以給出40μl細胞培養基總體積中每孔1500個細胞，並且在旋轉培養箱中在37℃、90%相對濕度以及5% CO2下培養過夜。然後藉由兩個測定方案A或B中的一種測試該等化合物： The MDA-MB-468 cells were cultured in a cell culture medium consisting of DMEM (Duller's modified Eagle's medium # D6546), 10% (v / v) fetal bovine serum, and 1% (v / v) L -Composition of glutamine. After harvesting, cells were distributed to black 384-well Costar plates (# 3712, Corning) to give 1500 cells per well in a total volume of 40 μl of cell culture medium, and in a rotary incubator at 37 ° C, 90% Incubate overnight at relative humidity and 5% CO2. These compounds are then tested by one of two assay schemes A or B:

方案A：Option A:

然後將該等細胞板在37℃下培養2小時，之後藉由添加在PBS/A(1.2%最終濃度)中的20μl 3.7%甲醛進行固定，隨後 是40分鐘室溫培養，並且然後使用BioTek Elx406洗板機用150μl PBS/A(磷酸鹽緩衝鹽水)洗滌2x。使細胞滲透化並且將細胞在室溫下用20μl的測定緩衝液(在PBS/A中的0.5%吐溫20+1%奶粉)阻斷1小時，並且然後用50μl PBS/A洗滌1x。將初始的磷-AKT(絲胺酸473)736E11兔單克隆抗體(#3787，細胞傳訊科技公司(Cell Signaling Technology))以1：500稀釋於測定緩衝液中，每孔添加20μl，並將該等板在4℃下培養過夜。將細胞板用200μl PBS/T(含有0.05%吐溫20的磷酸鹽緩衝鹽水)洗滌3x，然後向每孔中添加20μl在Alexa Fluor® 488山羊抗兔IgG二級抗體(#A11008，分子探針公司，生命科技公司)的測定緩衝液中的1：1000稀釋液，以及Hoechst 33342的1：5000稀釋液。在室溫下培養2小時後，將板用200μl PBS/T洗滌3x，並每孔添加40μl PBS/A。 The iso-cell plate was then incubated at 37 ° C for 2 hours, after which it was fixed by adding 20 μl 3.7% formaldehyde in PBS / A (1.2% final concentration), followed by 40 minutes incubation at room temperature, and then using BioTek Elx406 The plate washer was washed 2 × with 150 μl PBS / A (phosphate buffered saline). The cells were permeabilized and the cells were blocked with 20 μl of assay buffer (0.5% Tween 20 + 1% milk powder in PBS / A) for 1 hour at room temperature, and then washed 1 × with 50 μl of PBS / A. The initial phosphorus-AKT (serine 473) 736E11 rabbit monoclonal antibody (# 3787, Cell Signaling Technology) was diluted 1: 500 in assay buffer, 20 μl per well was added, and this The plates were incubated overnight at 4 ° C. The cell plate was washed 3x with 200 μl PBS / T (phosphate buffered saline containing 0.05% Tween 20), and then 20 μl of Alexa Fluor® 488 goat anti-rabbit IgG secondary antibody (# A11008, molecular probe) was added to each well Company, Life Science and Technology Co., Ltd.) 1: 1000 dilution in the assay buffer, and 1: 5000 dilution in Hoechst 33342. After incubating at room temperature for 2 hours, the plate was washed 3 × with 200 μl PBS / T, and 40 μl PBS / A was added per well.

將染色的細胞板用黑色密封件覆蓋，並且然後在Acumen(TTP萊伯泰科公司(TTP Labtech))讀板器上讀數。主要通道(綠色螢光，488nm)用於設置最大/最小截止的強度設置以允許染色的每週變化，並且將‘AKT+：物體數量(No)’數據用於分析。分析數據並且使用Genedata Screener®軟體計算IC50。 The stained cell plate was covered with a black seal, and then read on an Acumen (TTP Labtech) plate reader. The main channel (green fluorescence, 488nm) is used to set the intensity setting of the maximum / minimum cutoff to allow weekly changes in staining, and the 'AKT +: number of objects (No)' data is used for analysis. Analyze the data and calculate IC50 using Genedata Screener® software.

方案B：Option B:

然後將該等細胞板在37℃下培養2小時，之後藉由添加在PBS/A(1.2%最終濃度)中的20μl 3.7%甲醛進行固定，隨後是30分鐘室溫培養，並且然後使用BioTek Elx406洗板機用150μl PBS/A洗滌2x。使細胞滲透化並且將細胞在室溫下用20μl的測定緩衝液(在PBS/A中的0.1% Triton X-100+1% BSA)阻斷1h，並 且然後用50μl PBS/A洗滌1x。將初始的磷-AKT(絲胺酸473)D9E Xp®兔單克隆抗體(#4060，細胞傳訊科技公司(Cell Signaling Technology))以1：200稀釋於測定緩衝液中，每孔添加20μl，並將該等板在4℃下培養過夜。將細胞板用200μl PBS/T洗滌3x，然後向每孔中添加20μl的於Alexa Fluor® 488山羊抗兔IgG二級抗體(#A11008，分子探針公司，生命科技公司)的測定緩衝液中的1：750稀釋液，以及Hoechst 33342的1：5000稀釋液。在室溫下培養1小時後，將板用200μl PBS/T洗滌3x，並將40μl PBS w/o Ca、Mg以及Na Bicarb(Gibco #14190-094)添加至每孔中。 The iso-cell plate was then incubated at 37 ° C for 2 hours, after which it was fixed by adding 20 μl 3.7% formaldehyde in PBS / A (1.2% final concentration), followed by 30-minute incubation at room temperature, and then using BioTek Elx406 The plate washer was washed 2x with 150 μl PBS / A. The cells were permeabilized and the cells were blocked with 20 μl of assay buffer (0.1% Triton X-100 + 1% BSA in PBS / A) at room temperature for 1 h, and then washed 1 × with 50 μl of PBS / A. The initial phosphorus-AKT (serine 473) D9E Xp® rabbit monoclonal antibody (# 4060, Cell Signaling Technology) was diluted 1: 200 in assay buffer, 20 μl per well was added, and The plates were incubated overnight at 4 ° C. The cell plate was washed 3x with 200 μl PBS / T, and then 20 μl of the assay buffer in Alexa Fluor® 488 goat anti-rabbit IgG secondary antibody (# A11008, Molecular Probe, Life Science and Technology) was added to each well. 1: 750 dilution, and Hoechst 33342's 1: 5000 dilution. After incubating for 1 hour at room temperature, the plate was washed 3x with 200 μl PBS / T, and 40 μl PBS w / o Ca, Mg, and Na Bicarb (Gibco # 14190-094) were added to each well.

將染色的細胞板用黑色密封件覆蓋，並且然後在細胞透視成像平臺(Cell Insight imaging platform)(賽默科技公司)上用10x物鏡讀數。使用主要通道(Hoechst藍色螢光405nM，BGRFR_386_23)自動對焦並計數事件數目(這將提供關於所測試的化合物的細胞毒性的資訊)。第二通道(綠色488nM，BGRFR_485_20)測量pAKT染色。分析數據並且使用Genedata Screener®軟體計算IC50。 The stained cell plate was covered with a black seal, and then read with a 10x objective lens on a Cell Insight imaging platform (Thermo Technologies). Use the main channel (Hoechst blue fluorescent 405nM, BGRFR_386_23) to autofocus and count the number of events (this will provide information about the cytotoxicity of the compounds tested). The second channel (green 488nM, BGRFR_485_20) measures pAKT staining. Analysis of the data and IC50 was calculated using Genedata Screener ® software.

測定e)：DNAPK細胞效價 Measurement e): DNAPK cell titer

化合物處理：Compound treatment:

使用Echo 555聲學分配器(Labcyte IncTM)，將用於DNAPK細胞ELISA測定的所有化合物或DMSO(二甲亞碸)從包含100%(v/v)DMSO或100% DMSO中10mM化合物的來源板直接分配進測定板。將10mM化合物原料使用固定-尖96-頭安捷倫V製備型液體處理器(fixed-tip 96-head Agilent VPrep liquid handler) (加利福尼亞聖克拉拉安捷倫科技公司(Agilent Technologies))以1：100稀釋以給出四種中間體稀釋液(10mM、100μM、1μM、10nM)。藉由Echo將該中間板用於將化合物和DMSO直接分配至具有12點劑量範圍(30μM、10μM、3.125μM、1.25μM、0.3μM、0.1μM、0.03125μM、0.0125μM、0.003μM、0.001μM、0.0003125μM、0.00003μM)的細胞板中計算化合物IC50，其中測定中總DMSO濃度係0.3%。 Using an Echo 555 acoustic dispenser (Labcyte Inc ™ ), all compounds or DMSO (dimethyl sulfoxide) used for DNAPK cell ELISA assays were taken from a source plate containing 10 mM compounds in 100% (v / v) DMSO or 100% DMSO Distribute directly into the measuring plate. The 10 mM compound raw material was diluted 1: 100 using a fixed-tip 96-head Agilent VPrep liquid handler (Agilent Technologies, Santa Clara, California) to give Four intermediate dilutions (10 mM, 100 μM, 1 μM, 10 nM) were produced. The intermediate plate is used by Echo to dispense compounds and DMSO directly to have a 12-point dose range (30 μM, 10 μM, 3.125 μM, 1.25 μM, 0.3 μM, 0.1 μM, 0.03125 μM, 0.0125 μM, 0.003 μM, 0.001 μM, calculating compound IC 50 0.0003125μM, 0.00003μM) of the cell plate, wherein the total concentration of DMSO-based assay of 0.3%.

方法細節：Method details:

在HT29結腸直腸癌細胞系中進行DNA-PK細胞ELISA測定。將HT29細胞在細胞培養基中培養，該細胞培養基由MEM(最低必需培養基Eagle Sigma # M2279)、10%(v/v)胎牛血清和1%(v/v)200mM L-穀胺醯胺組成。收穫之後，將細胞分配至黑色384-孔Costar板(#3712，康寧公司(Corning))以給出40μl細胞培養基總體積中每孔15,000個細胞，並且在旋轉培養箱中在37℃、90%相對濕度以及5% CO2下培養過夜。將Greiner 781077全黑色高結合的384孔ELISA板用PBS中的0.5μg/mL DNA-PK抗體(Abcam # ab1832)在4℃下包被過夜。第二天，將Greiner ELISA板用PBS-T洗滌3x，並用3% BSA/PBS封閉約2小時，然後用PBS-T再洗滌3x。使用Labcyte Echo 555聲學分配器將測試化合物和參比對照組直接計量加入至細胞板。然後將細胞板在37℃下培養1小時，之後接受8Gy的輻射劑量(XRAD 320，台高度65)。將細胞再培養1小時，之後去除細胞培養基。以25μl/孔分配裂解緩衝液(添加有蛋白酶抑制劑混合物片劑的內部製劑，Roche # 04 693 116 001)，並且將板在4℃下培養15-20分鐘。使用CyBio Felix液體處理平臺將細胞裂解液(20μl/孔)轉移到DNA-PK抗體包被的ELISA板上，並將ELISA板在4℃下培養過夜。第二天，將ELISA板用PBS-T洗滌3x，並以20μl/孔分配內部pS2056-DNA-PK抗體(0.5μg/mL在3% BSA/PBS中)。將板與抗體在室溫(RT)下溫育1.5小時，之後用PBS-T洗滌3x。以20μl/孔分配山羊抗兔HRP二級抗體(1：2000稀釋於3% BSA/PBS中；細胞傳訊公司(Cell Signaling)# 7074)，並將板在室溫下培養1小時，然後用PBS-T洗滌3x。以20μl/孔分配QuantaBlu工作底物溶液(賽默飛科技公司(Thermo Scientific)# 15169，根據製造商的說明書製備)，並將板在室溫下培養1小時，然後用套組(賽默飛科技公司#15169)中提供的QuantaBlu終止溶液再分配20μl/孔。使用珀金埃爾默公司(PerkinElmer)EnVision讀板器測定單獨孔的螢光強度。分析數據並且使用Genedata Screener®軟體計算IC50。 DNA-PK cell ELISA assay was performed in HT29 colorectal cancer cell line. HT29 cells were cultured in a cell culture medium consisting of MEM (Minimal Essential Medium Eagle Sigma # M2279), 10% (v / v) fetal bovine serum, and 1% (v / v) 200 mM L-glutamine . After harvesting, cells were distributed to black 384-well Costar plates (# 3712, Corning) to give 15,000 cells per well in a total volume of 40 μl of cell culture medium, and in a rotary incubator at 37 ° C, 90% Incubate overnight at relative humidity and 5% CO 2 . Greiner 781077 all black high binding 384-well ELISA plates were coated with 0.5 μg / mL DNA-PK antibody (Abcam # ab1832) in PBS at 4 ° C overnight. The next day, the Greiner ELISA plate was washed 3x with PBS-T and blocked with 3% BSA / PBS for about 2 hours, and then washed with PBS-T for another 3x. The Labcyte Echo 555 acoustic dispenser was used to meter the test compound and the reference control group directly into the cell plate. The cell plate was then incubated at 37 ° C for 1 hour, after which it received a radiation dose of 8 Gy (XRAD 320, stage height 65). The cells were cultured for another hour, after which the cell culture medium was removed. The lysis buffer (internal preparation with the addition of protease inhibitor cocktail tablets, Roche # 04 693 116 001) was dispensed at 25 μl / well, and the plate was incubated at 4 ° C for 15-20 minutes. The cell lysate (20 μl / well) was transferred to a DNA-PK antibody-coated ELISA plate using CyBio Felix liquid processing platform, and the ELISA plate was incubated at 4 ° C overnight. The next day, the ELISA plate was washed 3x with PBS-T and the internal pS2056-DNA-PK antibody (0.5 μg / mL in 3% BSA / PBS) was dispensed at 20 μl / well. The plate was incubated with antibody at room temperature (RT) for 1.5 hours, after which it was washed 3x with PBS-T. Dispense goat anti-rabbit HRP secondary antibody (diluted 1: 2000 in 3% BSA / PBS; Cell Signaling # 7074) at 20 μl / well, and incubate the plate at room temperature for 1 hour, then use PBS -T wash 3x. QuantaBlu working substrate solution (Thermo Scientific # 15169, prepared according to the manufacturer's instructions) was dispensed at 20 μl / well, and the plate was incubated at room temperature for 1 hour, and then set (thermofei The QuantaBlu stop solution provided in Technology Company # 15169) was dispensed by another 20 μl / well. PerkinElmer EnVision plate reader was used to measure the fluorescence intensity of individual wells. Analyze the data and calculate IC50 using Genedata Screener® software.

§使用測定d)方案A得到的結果 §Results obtained by using measurement d) Scheme A

*使用測定d)方案B得到的結果 * Results obtained using measurement d) Scheme B

表5示出在測定a)至e)中針對CN 102399218 A和CN 102372711 A的某些化合物的比較性數據。 Table 5 shows comparative data for certain compounds of CN 102399218 A and CN 102372711 A in assays a) to e).

§使用測定d)方案A得到的結果 §Results obtained by using measurement d) Scheme A

Claims (15)

一種具有化學式 (I)之化合物： 或其藥學上可接受的鹽，其中： R 1 係四氫哌喃-3-基或3,3-二甲基四氫哌喃-4-基； R 2 係甲基或氫； R 3 係氫或氟； R 4 係氫或氟；並且 R 5 係甲基或氫。 A compound of formula (I) : Or a pharmaceutically acceptable salt thereof, wherein: R 1 is tetrahydropiperan-3-yl or 3,3-dimethyltetrahydropiperan-4-yl; R 2 is methyl or hydrogen; R 3 is Hydrogen or fluorine; R 4 is hydrogen or fluorine; and R 5 is methyl or hydrogen. 如申請專利範圍第1項所述之具有化學式 (I)之化合物或其藥學上可接受的鹽，其中 R 1 係四氫哌喃-3-基。 The compound of formula (I) or a pharmaceutically acceptable salt thereof as described in item 1 of the patent application scope, wherein R 1 is tetrahydropiperan-3-yl. 如申請專利範圍第1項所述之具有化學式 (I)之化合物或其藥學上可接受的鹽，其中 R 1 係3,3-二甲基四氫哌喃-4-基。 The compound of formula (I) or a pharmaceutically acceptable salt thereof as described in item 1 of the patent scope, wherein R 1 is 3,3-dimethyltetrahydropiperan-4-yl. 如前述申請專利範圍中任一項所述之具有化學式 (I)之化合物或其藥學上可接受的鹽，其中 R 2 係甲基。 The compound of formula (I) or a pharmaceutically acceptable salt thereof as described in any of the foregoing patent applications, wherein R 2 is methyl. 如前述申請專利範圍中任一項所述之具有化學式 (I)之化合物或其藥學上可接受的鹽，其中 R 3 係氫。 The compound of formula (I) or a pharmaceutically acceptable salt thereof as described in any of the foregoing patent applications, wherein R 3 is hydrogen. 如前述申請專利範圍中任一項所述之具有化學式 (I)之化合物或其藥學上可接受的鹽，其中 R 4 係氫。 The compound of formula (I) or a pharmaceutically acceptable salt thereof as described in any of the foregoing patent applications, wherein R 4 is hydrogen. 如申請專利範圍第1項所述之具有化學式 (I)之化合物或其藥學上可接受的鹽，其中： R 1 係四氫哌喃-3-基； R 2 係甲基或氫； R 3 係氫或氟； R 4 係氫或氟；並且 R 5 係甲基。 The compound of formula (I) or a pharmaceutically acceptable salt thereof as described in item 1 of the patent scope, wherein: R 1 is tetrahydropiperan-3-yl; R 2 is methyl or hydrogen; R 3 Is hydrogen or fluorine; R 4 is hydrogen or fluorine; and R 5 is methyl. 如申請專利範圍第1項所述之具有化學式 (I)之化合物或其藥學上可接受的鹽，其中該化合物選自：( R)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮；( S)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮；( S)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮；( R)-7-氟-8-(2-氟-6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮；( S)-7-氟-8-(2-氟-6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮；( R)-8-(2-氟-6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮；( S)-8-(2-氟-6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3- 基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮；( R)-7-氟-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1-(四氫-2H-哌喃-3-基)-1H-咪唑并[4,5-c]喹啉-2(3H)-酮；外消旋-1-(3,3-二甲基四氫-2H-哌喃-4-基)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮；( R)-1-(3,3-二甲基四氫-2H-哌喃-4-基)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮；( S)-1-(3,3-二甲基四氫-2H-哌喃-4-基)-8-(6-(甲氧基甲基)吡啶-3-基)-3-甲基-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮；以及( S)-8-(6-(甲氧基甲基)吡啶-3-基)-1-(四氫-2H-哌喃-3-基)-1,3-二氫-2H-咪唑并[4,5-c]喹啉-2-酮。 The compound of formula (I) or a pharmaceutically acceptable salt thereof as described in item 1 of the patent application scope, wherein the compound is selected from: ( R ) -8- (6- (methoxymethyl) pyridine- 3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5-c] quinolin-2 (3H) -one; ( S ) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5- c] Quinolin-2 (3H) -one; ( S ) -7-fluoro-8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro- 2H-piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one; ( R ) -7-fluoro-8- (2-fluoro- 6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5-c] quinoline -2 (3H) -one; ( S ) -7-fluoro-8- (2-fluoro-6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro- 2H-piperan-3-yl) -1H-imidazo [4,5-c] quinolin-2 (3H) -one; ( R ) -8- (2-fluoro-6- (methoxymethyl ) Pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5-c] quinolin-2 (3H) -one; ( S ) -8- (2-fluoro-6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H- Imidazo [4,5-c] quinolin-2 (3H) -one; ( R ) -7-fluoro -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1- (tetrahydro-2H-piperan-3-yl) -1H-imidazo [4,5- c] quinoline-2 (3H) -one; racemic-1- (3,3-dimethyltetrahydro-2H-piperan-4-yl) -8- (6- (methoxymethyl ) Pyridin-3-yl) -3-methyl-1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one; ( R ) -1- (3,3-di Methyltetrahydro-2H-piperan-4-yl) -8- (6- (methoxymethyl) pyridin-3-yl) -3-methyl-1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one; ( S ) -1- (3,3-dimethyltetrahydro-2H-piperan-4-yl) -8- (6- (methoxy Methyl) pyridin-3-yl) -3-methyl-1,3-dihydro-2H-imidazo [4,5-c] quinolin-2-one; and ( S ) -8- (6- (Methoxymethyl) pyridin-3-yl) -1- (tetrahydro-2H-piperan-3-yl) -1,3-dihydro-2H-imidazo [4,5-c] quinoline -2-one. 一種醫藥組成物，該醫藥組成物包括如申請專利範圍第1至8項中任一項所述之具有化學式 (I)之化合物或其藥學上可接受的鹽，以及至少一種藥學上可接受的賦形劑。 A pharmaceutical composition comprising the compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 8 of the patent application, and at least one pharmaceutically acceptable excipient. 如申請專利範圍第1至8項中任一項所述之具有化學式 (I)之化合物或其藥學上可接受的鹽，用於在療法中使用。 The compound of formula (I) or a pharmaceutically acceptable salt thereof as described in any one of claims 1 to 8 of the patent application scope is used in therapy. 如申請專利範圍第1至8項中任一項所述之具有化學式 (I)之化合物或其藥學上可接受的鹽，用於在癌症的治療中使用。 The compound of formula (I) or a pharmaceutically acceptable salt thereof as described in any one of claims 1 to 8 of the patent application scope is used in the treatment of cancer. 一種如申請專利範圍第11項所述之、用於在癌症的治療中使用的具有化學式 (I)之化合物或其藥學上可接受的鹽，其中該具有化學式 (I)之化合物與放射療法被同時地、分別地或順序地給予。 A compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer as described in item 11 of the patent application scope, wherein the compound of formula (I) and radiation therapy are Give at the same time, separately or sequentially. 一種如申請專利範圍第11項所述之、用於在癌症的治療中使用 的具有化學式 (I)之化合物或其藥學上可接受的鹽，其中該具有化學式 (I)之化合物與至少一種另外的抗腫瘤物質被同時地、分別地或順序地給予，該另外的抗腫瘤物質選自多柔比星、伊立替康、托泊替康、依託泊苷、絲裂黴素、苯達莫司汀、苯丁酸氮芥、環磷醯胺、異環磷醯胺、卡莫司汀、美法侖以及博萊黴素。 A compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer as described in item 11 of the patent application scope, wherein the compound of formula (I) and at least one additional The anti-tumor substance is administered simultaneously, separately or sequentially, the additional anti-tumor substance is selected from doxorubicin, irinotecan, topotecan, etoposide, mitomycin, bendamus Ting, chlorambucil, cyclophosphamide, ifosfamide, carmustine, melphalan and bleomycin. 如申請專利範圍第1至8項中任一項所述之具有化學式 (I)之化合物或其藥學上可接受的鹽在生產用於治療癌症的藥物中之用途。 The use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as described in any one of items 1 to 8 of the patent application scope in the production of a medicament for treating cancer. 一種用於在需要此類治療的溫血動物中治療癌症之方法，該方法包括向所述溫血動物給予治療有效量的如申請專利範圍第1至8項中任一項所述之具有化學式 (I)之化合物或其藥學上可接受的鹽。 A method for treating cancer in a warm-blooded animal in need of such treatment, the method comprising administering to the warm-blooded animal a therapeutically effective amount of a chemical formula as described in any one of claims 1 to 8 The compound of (I) or a pharmaceutically acceptable salt thereof.
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* Cited by examiner, † Cited by third party
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CN115232122A (en) * 2021-04-23 2022-10-25 石药集团中奇制药技术（石家庄）有限公司 Alkyne compound and preparation and application thereof

Families Citing this family (5)

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CN115003672A (en) * 2020-01-09 2022-09-02 南京明德新药研发有限公司 Quinoline imidazole compound and application thereof
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EP3992191A1 (en) 2020-11-03 2022-05-04 Deutsches Krebsforschungszentrum Imidazo[4,5-c]quinoline compounds and their use as atm kinase inhibitors
WO2022128833A1 (en) 2020-12-15 2022-06-23 Merck Patent Gmbh Solid transition metal-ligand complexes

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* Cited by examiner, † Cited by third party
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TR199801530T2 (en) 1996-02-13 1998-11-23 Zeneca Limited Quinazoline derivatives as VEGF inhibitors.
JP4464466B2 (en) 1996-03-05 2010-05-19 アストラゼネカ・ユーケイ・リミテッド 4-anilinoquinazoline derivatives
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CN102372711B (en) 2010-08-18 2014-09-17 山东轩竹医药科技有限公司 Imidazo quinoline PI3K and mTOR (mammalian target of rapamycin) dual inhibitor
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Cited By (1)

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