TW201518301A - Spiropyrazolopyridine derivatives and uses thereof - Google Patents

Spiropyrazolopyridine derivatives and uses thereof Download PDF

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TW201518301A
TW201518301A TW103113267A TW103113267A TW201518301A TW 201518301 A TW201518301 A TW 201518301A TW 103113267 A TW103113267 A TW 103113267A TW 103113267 A TW103113267 A TW 103113267A TW 201518301 A TW201518301 A TW 201518301A
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compound
pharmaceutically acceptable
acceptable salt
pyridine
dione
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TW103113267A
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Wai Ling Chan
Mei Ding
Bin Zou
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Novartis Ag
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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
    • C07D471/12—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 three hetero rings
    • C07D471/20—Spiro-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses

Abstract

A compound of Formula (I) is provided that has been shown to be useful for treating a disease caused by a viral infection: wherein R1, R2, R3, R4, R5, R6, R7, R8, X1, X2, X3 and X4, are as defined herein.

Description

螺吡唑并吡啶衍生物及其用途 Spiropyrazolopyridine derivatives and uses thereof

本申請案主張2013年4月11日申請之國外新加坡專利申請案第201302813-9號之優先權，其內容以全文引用的方式併入本文中。 The present application claims priority to Japanese Patent Application No. 201302813-9, filed on Apr. 11, 2013, the content of which is incorporated herein in its entirety.

本發明係關於螺吡唑并吡啶衍生物、其醫藥調配物及其用於治療病毒感染、尤其為由登革熱病毒(dengue virus)引起之病毒感染的用途。 The present invention relates to spiropyrazolopyridine derivatives, pharmaceutical formulations thereof and their use for the treatment of viral infections, in particular viral infections caused by dengue virus.

登革熱(Dengue fever)係由屬於黃病毒科(Flaviviridae)家族之四種登革熱病毒血清型DEN-1、DEN-2、DEN-3及DEN-4之一引起的發熱性疾病。此病毒主要由埃及斑紋(Aedes aegypti)(一種以人類為食之蚊子)傳播給人類。 Dengue fever is a febrile disease caused by one of the four dengue virus serotypes DEN-1, DEN-2, DEN-3, and DEN-4 belonging to the Flaviviridae family. The virus is mainly transmitted to humans by the Egyptian ( Aedes aegypti ), a mosquito that feeds on humans.

感染產生多種臨床表現，由較輕度之流感樣症狀至更嚴重且有時致命之出血性疾病。典型症狀包括發熱、嚴重頭痛、肌肉及關節痛以及發疹。更嚴重之疾病形式係登革出血熱(dengue haemorrhagic fever，DHF)及登革休克症候群(dengue shock syndrome，DSS)。根據WHO，DHF存在四種主要的臨床表現：(1)高熱、(2)出血現象、(3)血小板減少及(4)血漿滲漏。DSS定義為DHF伴有弱而速之脈搏及脈壓窄或低血壓伴有感冒、皮膚濕冷及躁動。藉由早期偵測及介入可使DHF之嚴重性降低，但休克之個體存在高死亡風險。 Infections produce a variety of clinical manifestations, ranging from milder flu-like symptoms to more severe and sometimes fatal bleeding disorders. Typical symptoms include fever, severe headache, muscle and joint pain, and rash. More serious forms of disease are dengue haemorrhagic fever (DHF) and dengue shock syndrome (DSS). According to WHO, there are four major clinical manifestations of DHF: (1) hyperthermia, (2) hemorrhage, (3) thrombocytopenia, and (4) plasma leakage. DSS is defined as DHF with weak and rapid pulse and narrow pulse or low blood pressure with cold, cold skin and irritability. The severity of DHF can be reduced by early detection and intervention, but individuals with shock have a high risk of death.

登革熱係熱帶地區之地方病，尤其在亞洲、非洲及拉丁美洲，且估計有25億人生活在存在感染風險的區域。每年大約有4000萬登革熱病例及數十萬DHF病例。在新加坡，2005年的流行病導致12000例以上之登革熱。 Dengue is a local disease in the tropics, especially in Asia, Africa and Latin America, and an estimated 2.5 billion people live in areas where there is a risk of infection. There are approximately 40 million cases of dengue fever and hundreds of thousands of DHF cases each year. In Singapore, the epidemic in 2005 caused more than 12,000 cases of dengue fever.

儘管定期發作，但先前受感染之人體仍易受感染，因為存在四種不同之登革熱病毒血清型且具有該等血清型之一之感染僅對彼血清型提供免疫。咸信在患有繼發性登革熱感染之個體中更易於發生DHF。正在尋求對於登革熱、DHF及DSS之有效治療。 Despite regular onset, previously infected humans are still susceptible to infection because there are four different dengue virus serotypes and infection with one of these serotypes only immunizes the serotype. Xianxin is more prone to DHF in individuals with secondary dengue infection. Effective treatment for dengue, DHF and DSS is being sought.

黃病毒基因體係長度約11kb之單股、正義RNA。其含有側接單一開讀框之5'及3'未轉譯區(UTR)。開讀框編碼與轉譯同時或在轉譯後由病毒及細胞蛋白酶加工為三個結構蛋白(蛋白殼[C]、前膜[prM]及包膜[E])及七個非結構蛋白(NS1、NS2A、NS2B、NS3、NS4A、NS4B及NS5)之長多聚蛋白。在病毒進入及組裝中涉及結構蛋白。在病毒RNA複製、固有免疫反應逃避及病毒組裝中涉及非結構蛋白。 The flavivirus gene system is a single strand, sense RNA of approximately 11 kb in length. It contains 5' and 3' untranslated regions (UTR) flanked by a single open reading frame. The open reading frame coding and translation are processed by virus and cellular protease into three structural proteins (protein shell [C], anterior membrane [prM] and envelope [E]) and seven non-structural proteins (NS1). Long polyproteins of NS2A, NS2B, NS3, NS4A, NS4B and NS5). Structural proteins are involved in viral entry and assembly. Non-structural proteins are involved in viral RNA replication, innate immune response evasion, and viral assembly.

醣蛋白NS1在病毒RNA複製之早期步驟中對病毒RNA複製具有作用。NS3充當具有輔因子NS2B、RNA三磷酸酶及RNA解旋酶之病毒絲胺酸蛋白酶。NS5充當甲基轉移酶及RNA依賴性RNA聚合酶(RdRp)。關於小的疏水性蛋白NS2A、NS4A及NS4B，該等跨膜蛋白將病毒複製複合物錨定至內質網(ER)膜。 Glycoprotein NS1 has an effect on viral RNA replication in the early steps of viral RNA replication. NS3 acts as a viral serine protease with the cofactor NS2B, RNA triphosphatase and RNA helicase. NS5 acts as a methyltransferase and an RNA-dependent RNA polymerase (RdRp). With respect to the small hydrophobic proteins NS2A, NS4A and NS4B, these transmembrane proteins anchor the viral replication complex to the endoplasmic reticulum (ER) membrane.

黃熱病毒(黃熱病毒，YFV)、西尼羅病毒(West Nile virus，WNV)、日本腦炎病毒(Japanese encephalitis virus，JEV)、布氏病毒(Powassan virus，POWV)、蜱傳腦炎病毒、庫京病毒(Kunjin virus)、墨累谷腦炎(Murray Valley encephalitis)、聖路易腦炎(St Louis encephalitis)、鄂木斯克出血性熱病毒(Omsk hemorrhagic fever virus)、牛病毒性腹瀉病毒、寨卡病毒(Zika virus)及C型肝炎病毒(Hepatitis C virus，HCV)亦屬於黃病毒科家族。 Yellow fever virus (Yellow fever virus, YFV), West Nile virus (WNV), Japanese encephalitis virus (JEV), Powassan virus (POWV), tick-borne encephalitis virus , Kunjin virus, Murray Valley encephalitis, St Louis encephalitis, Omsk hemorrhagic fever virus, bovine viral diarrhea virus, Zika virus and Hepatitis C virus (HCV) also belong to the family Flaviviridae.

WNV可無症狀，或在某些個體中可引起流感樣症狀。在某些情形下，其引起神經障礙、腦炎，且在嚴重情形下可導致死亡。WNV亦由蚊子來傳播。YFV係另一種蚊媒病毒，其可引起受感染個體之嚴重症狀。JEV亦由蚊子傳播且無症狀或引起流感樣症狀，某些情形下發展成腦炎。疾病之急性腦炎階段具有抽搐、頸強直及其他症狀特徵。HCV係由血液接觸傳播之血液傳染病毒。在疾病之初始(急性)階段，大多數個體不展現任何症狀。即使在慢性階段(亦即，疾病持續6個月以上)，個體之間之症狀嚴重程度亦可不同。長遠而言，某些受感染人體可發展成肝硬化及肝癌。目前對於HCV之治療涉及干擾素α與利巴韋林(ribavirin，一種抗病毒藥物)之組合。亦正在尋求對於由該等黃病毒科病毒引起之感染的有效治療。 WNV can be asymptomatic or can cause flu-like symptoms in some individuals. In some cases, it causes neurological disorders, encephalitis, and in severe cases can lead to death. WNV is also spread by mosquitoes. YFV is another mosquito-borne virus that causes severe symptoms in infected individuals. JEV is also transmitted by mosquitoes and is asymptomatic or causes flu-like symptoms, and in some cases develops into encephalitis. The acute encephalitis stage of the disease has convulsions, neck stiffness and other symptomatic features. HCV is a blood-borne virus that is transmitted by blood contact. Most individuals do not exhibit any symptoms during the initial (acute) phase of the disease. Even in the chronic phase (ie, the disease lasts for more than 6 months), the severity of symptoms between individuals can vary. In the long run, some infected humans can develop into cirrhosis and liver cancer. Current treatments for HCV involve a combination of interferon alpha and ribavirin, an antiviral drug. Effective treatment for infections caused by such Flaviviridae viruses is also being sought.

現已令人驚訝地發現，如本文所揭示之螺吡唑并吡啶衍生物適用於治療病毒感染，諸如由黃病毒科家族之病毒(尤其為登革熱病毒及黃熱病毒)引起之彼等病毒感染。 It has now surprisingly been found that the spiropyrazolopyridine derivatives as disclosed herein are suitable for the treatment of viral infections, such as those caused by viruses of the Flaviviridae family, in particular dengue virus and yellow fever virus. .

本文所述之化合物已展示適用於預防及/或治療病毒感染。 The compounds described herein have been shown to be useful for the prevention and/or treatment of viral infections.

本發明之一態樣提供式(I)化合物 One aspect of the invention provides a compound of formula (I)

其中 R1為F、Cl、Br、CF3、OCH3、(C1-C6)烷基或環烷基；R2為H、F、Cl、Br、(C1-C6)烷基或環烷基；或R1與R2連同其所連接之芳族碳原子一起形成稠合1,3-間二氧雜環戊烯基；R3為H、(C1-C6)烷基或環烷基；R4為H、(C1-C6)烷基、環烷基或-C(=O)-NH2；R5為H、(C1-C6)烷基、環烷基或苄基；R6為H、(C1-C6)烷基或環烷基；R7為H、F、Cl或OCH3；R8為F、Cl、Br、CF3、-OCHF2、OCF3、OCH3或(C1-C6)烷基；X1為CH或N；X2為CH或N；X3為CH或N；X4為=O或H2；R1、R2、R3、R4、R5、R6、R7及R8各自視情況獨立地經一或多個獨立的R300取代基取代；R300係選自由H、(C1-C6)烷基、環烷基、羥基、胺基、F、Cl、Br、苯基、雜環及雜芳基組成之群；或其醫藥學上可接受之鹽。 Wherein R 1 is F, Cl, Br, CF 3 , OCH 3 , (C 1 -C 6 )alkyl or cycloalkyl; and R 2 is H, F, Cl, Br, (C 1 -C 6 )alkyl Or a cycloalkyl group; or R 1 and R 2 together with the aromatic carbon atom to which they are attached form a fused 1,3-dioxolyl; R 3 is H, (C 1 -C 6 ) alkane Or a cycloalkyl group; R 4 is H, (C 1 -C 6 )alkyl, cycloalkyl or -C(=O)-NH 2 ; R 5 is H, (C 1 -C 6 )alkyl, a cycloalkyl or benzyl group; R 6 is H, (C 1 -C 6 )alkyl or cycloalkyl; R 7 is H, F, Cl or OCH 3 ; R 8 is F, Cl, Br, CF 3 , -OCHF 2 , OCF 3 , OCH 3 or (C 1 -C 6 )alkyl; X 1 is CH or N; X 2 is CH or N; X 3 is CH or N; X 4 is =O or H 2 ; R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 and R 8 are each optionally substituted by one or more independent R 300 substituents; R 300 is selected from H, (C) a group consisting of 1 -C 6 )alkyl, cycloalkyl, hydroxy, amine, F, Cl, Br, phenyl, heterocyclic and heteroaryl; or a pharmaceutically acceptable salt thereof.

在一實施例中，式(I)化合物或其醫藥學上可接受之鹽，其中R1為Cl。在另一實施例中，式(I)化合物或其醫藥學上可接受之鹽，其中R2為Cl或F。在又一實施例中，式(I)化合物或其醫藥學上可接受之鹽，其中R3為H或環丙基。 In one embodiment, a compound of formula acceptable (I) or a pharmaceutically acceptable salt thereof, wherein R 1 is Cl. In another embodiment, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R 2 is Cl or F. In yet another embodiment, a compound of formula acceptable (I) or a pharmaceutically acceptable salt thereof, wherein R 3 is H or cyclopropyl.

在另一實施例中，式(I)化合物或其醫藥學上可接受之鹽，其中R4為H。在又一實施例中，式(I)化合物或其醫藥學上可接受之鹽，其中R5為H、甲基或異丙基。在又一實施例中，式(I)化合物或其醫藥學上可接受之鹽，其中R6為H或甲基。 In another embodiment, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R 4 is H. In yet another embodiment, Formula (I) compound or a pharmaceutically acceptable salt thereof, wherein R 5 is H, methyl or isopropyl. In yet another embodiment, Formula (I) compound or a pharmaceutically acceptable salt thereof, wherein R 6 is H or methyl.

在另一實施例中，式(I)化合物或其醫藥學上可接受之鹽，其中R7為H或Cl。在又一實施例中，式(I)化合物或其醫藥學上可接受之鹽，R8為Cl、甲基、異丙基或第三丁基。 In another embodiment, a compound of formula acceptable (I) or a pharmaceutically acceptable salt thereof, wherein R 7 is H or Cl. In yet another embodiment, Formula (I) compound or a pharmaceutically acceptable salt, R 8 is Cl, methyl, isopropyl or tert-butyl.

在另一實施例中，式(I)化合物或其醫藥學上可接受之鹽，其中X1為CH。在又一實施例中，式(I)化合物或其醫藥學上可接受之鹽，其中X2為N。在又一實施例中，式(I)化合物或其醫藥學上可接受之鹽，其中X3為CH。在又一實施例中，式(I)化合物或其醫藥學上可接受之鹽，其中X4為=O。 In another embodiment, the pharmaceutically formula (I), or a pharmaceutically acceptable salt thereof, wherein X 1 is CH. In a further embodiment, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein X 2 is N. In yet another embodiment, Formula (I) compound or a pharmaceutically acceptable salt thereof, wherein X 3 is CH. In still another embodiment, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein X 4 is =0.

代表性式(I)化合物或其醫藥學上可接受之鹽呈現在下表1中： Representative compounds of formula (I) or pharmaceutically acceptable salts thereof are presented in Table 1 below:

其他代表性式(I)化合物或其醫藥學上可接受之鹽呈現於下表2中： Other representative compounds of formula (I) or pharmaceutically acceptable salts thereof are presented in Table 2 below:

受到特定關注之化合物或其醫藥學上可接受之鹽呈現於下表3中： Compounds of particular interest or pharmaceutically acceptable salts thereof are presented in Table 3 below:

受到特定關注之其他化合物或其醫藥學上可接受之鹽呈現於下 表4中： Other compounds of particular interest or pharmaceutically acceptable salts thereof are presented in Table 4 below:

本發明之另一態樣包括一種醫藥組合物，其包含包括上文所述實施例中任一者之式(I)化合物或其醫藥學上可接受之鹽及醫藥學上可接受之載劑或賦形劑。醫藥組合物可進一步包含至少一種其他醫藥劑。其他醫藥劑之實例包括(但不限於)西戈斯韋(Celgosivir)。 Another aspect of the invention includes a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, including any of the above-described embodiments Or an excipient. The pharmaceutical composition may further comprise at least one other pharmaceutical agent. Examples of other pharmaceutical agents include, but are not limited to, Celgosivir.

在本發明之又一態樣中，提供一種治療由病毒感染引起之疾病的方法，其包含向有此需要之個體(特定而言為人類)投與治療有效量之式(I)化合物(包括本文所述之任何實施例)或其醫藥學上可接受之鹽的步驟。在一特定適用實施例中，病毒感染由選自由登革熱病毒及黃熱病毒組成之群之病毒引起。在一更特定適用實施例中，病毒感染由登革熱病毒引起。化合物可以本文所述之醫藥組合物形式來投與。 In a further aspect of the invention, a method of treating a disease caused by a viral infection comprising administering to a subject in need thereof, in particular a human, a therapeutically effective amount of a compound of formula (I), Any of the embodiments described herein) or a pharmaceutically acceptable salt thereof. In a particularly suitable embodiment, the viral infection is caused by a virus selected from the group consisting of dengue virus and yellow fever virus. In a more specific suitable embodiment, the viral infection is caused by a dengue virus. The compounds can be administered in the form of a pharmaceutical composition as described herein.

本發明之另一態樣包括一種式(I)化合物(包含上文所述實施例中 之任一者)或其醫藥學上可接受之鹽，其係用作藥物(例如，使用式(I)化合物(包含上文所述實施例中之任一者)或其醫藥學上可接受之鹽製造用於治療由病毒感染引起之疾病的藥物)。在一特定適用實施例中，病毒感染由選自由登革熱病毒及黃熱病毒組成之群之病毒引起。在一更特定適用實施例中，病毒感染由登革熱病毒引起。本發明之另一態樣包括一種式(I)化合物(包含上文所述實施例中之任一者)或其醫藥學上可接受之鹽，其中化合物為NS4B抑制劑。 Another aspect of the invention includes a compound of formula (I) (including the examples described above) Or a pharmaceutically acceptable salt thereof for use as a medicament (for example, using a compound of formula (I), including any of the embodiments described above, or a pharmaceutically acceptable thereof The salt is used to manufacture a drug for treating a disease caused by a viral infection). In a particularly suitable embodiment, the viral infection is caused by a virus selected from the group consisting of dengue virus and yellow fever virus. In a more specific suitable embodiment, the viral infection is caused by a dengue virus. Another aspect of the invention comprises a compound of formula (I), including any of the above-described embodiments, or a pharmaceutically acceptable salt thereof, wherein the compound is an NS4B inhibitor.

定義definition

片語「治療有效量」意謂(i)治療或預防特定疾病、病狀或病症，(ii)減輕、改善或消除特定疾病、病狀或病症之一或多種症狀，或(iii)預防或延遲本文所述之特定疾病、病狀或病症之一或多種症狀發作的本發明化合物之量。 The phrase "therapeutically effective amount" means (i) treating or preventing a particular disease, condition or disorder, (ii) reducing, ameliorating or eliminating one or more symptoms of a particular disease, condition or condition, or (iii) preventing or An amount of a compound of the invention that delays the onset of one or more symptoms of a particular disease, condition or disorder described herein.

如本文所用，若個體將在生物學、醫學或在生活品質方面受益於治療，則此個體「需要」此治療(較佳為人類)。 As used herein, an individual "needs" the treatment (preferably a human) if the individual will benefit from treatment in biology, medicine, or quality of life.

如本文所用，術語「抑制(inhibit)」、「抑制(inhibition)」或「抑制(inhibiting)」係指減輕或遏止既定病狀、症狀或病症或疾病，或顯著減小生物活性或過程之基線活性。 As used herein, the terms "inhibit", "inhibition" or "inhibiting" mean reducing or arresting a given condition, symptom or condition or disease, or significantly reducing the baseline of biological activity or process. active.

如本文所用，術語「個體」係指動物。動物通常為哺乳動物。 個體亦係指例如靈長類動物(例如人類)、奶牛、綿羊、山羊、馬、狗、貓、兔、大鼠、小鼠、魚、鳥及其類似動物。在某些實施例中，個體為靈長類動物。又在其他實施例中，個體為人類。 As used herein, the term "individual" refers to an animal. Animals are usually mammals. An individual also refers to, for example, a primate (eg, a human), a cow, a sheep, a goat, a horse, a dog, a cat, a rabbit, a rat, a mouse, a fish, a bird, and the like. In certain embodiments, the individual is a primate. In still other embodiments, the individual is a human.

如本文所用，在一實施例中，術語「治療(treat)」、「治療(treating)」或「治療(treatment)」任何疾病或病症係指緩解疾病或病症(亦即，減緩或阻止或減慢疾病或其至少一種臨床症狀進展)。在另一實施例中，「治療」係指減輕或緩解至少一個身體參數，包括個體不可辨別之身體參數。在又一實施例中，「治療」係指在身體上(例如穩定可辨別之症狀)、生理上(例如穩定身體參數)或兩方面調節疾病或病症。在又一實施例中，「治療」係指預防或延遲疾病或病症的發作或發展或進展。 As used herein, in one embodiment, the term "treat", "treating" or "treatment" refers to alleviating a disease or condition (ie, slowing or preventing or reducing). Slow disease or progression of at least one of its clinical symptoms). In another embodiment, "treating" refers to alleviating or alleviating at least one physical parameter, including an individual's indistinguishable physical parameter. In yet another embodiment, "treating" refers to modulating a disease or condition either physically (eg, to stabilize a discernible symptom), physiologically (eg, to stabilize a body parameter), or both. In yet another embodiment, "treating" refers to preventing or delaying the onset or progression or progression of a disease or condition.

片語「醫藥學上可接受」表示物質或組合物須與構成調配物之其他成份及/或用其治療之哺乳動物在化學上及/或毒理學上相容。 The phrase "pharmaceutically acceptable" means that the substance or composition is chemically and/or toxicologically compatible with the other ingredients which comprise the formulation and/or the mammal to which it is treated.

術語「本發明之化合物」(除非另外明確規定)係指式(I)化合物及其鹽以及所有立體異構體(包括非對映異構體及對映異構體)、旋轉異構體、互變異構體及同位素標記化合物(包括氘取代)以及固有形成之部分(例如，多晶型物、溶劑合物及/或水合物)。為本發明之目的，溶劑合物及水合物一般認為係組合物。 The term "compound of the invention" (unless otherwise specifically indicated) refers to a compound of formula (I) and salts thereof, and all stereoisomers (including diastereomers and enantiomers), rotamers, Tautomers and isotopically labeled compounds (including deuterium substitutions) as well as intrinsically formed moieties (eg, polymorphs, solvates, and/or hydrates). For the purposes of the present invention, solvates and hydrates are generally considered to be compositions.

除非本文另外指示或上下文中明顯矛盾，否則本發明上下文中(尤其申請專利範圍上下文中)所用之術語「一(a)」、「一(an)」及「該(the)」及類似術語應解釋為涵蓋單數及複數形式。 The terms "a", "an" and "the" and the like are used in the context of the present invention (especially in the context of the claims) unless the context clearly indicates otherwise. Interpreted as covering both singular and plural forms.

如本文所用，術語「烷基」係指通式CnH2n+1之烴基。烷烴基團可為直鏈或分支鏈。舉例而言，術語「(C1-C6)烷基」係指含有1至6個碳原子之單價直鏈或分支鏈脂族基團(例如甲基、乙基、正丙基、異丙基、正丁基、異丁基、第二丁基、第三丁基、正戊基、1-甲基丁基、2-甲基丁基、3-甲基丁基、新戊基、3,3-二甲基丙基、己基、2- 甲基戊基及其類似基團)。類似地，烷氧基、醯基(例如，烷醯基)、烷胺基、二烷胺基及烷硫基之烷基部分(alkyl portion)(亦即，烷基部分(alkyl moiety))具有如上文相同之定義。 As used herein, the term "alkyl" means a hydrocarbyl group of formula C n H 2n + 1. The alkane group can be a straight or branched chain. For example, the term "(C 1 -C 6 )alkyl" refers to a monovalent straight or branched aliphatic group containing from 1 to 6 carbon atoms (eg, methyl, ethyl, n-propyl, isopropyl) Base, n-butyl, isobutyl, t-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, 3 , 3-dimethylpropyl, hexyl, 2-methylpentyl and the like. Similarly, an alkyl moiety (i.e., an alkyl moiety) having an alkoxy group, a fluorenyl group (e.g., an alkyl fluorenyl group), an alkylamino group, a dialkylamino group, and an alkylthio group has The same definition as above.

術語「環烷基」係指完全氫化且以單環形式存在之非芳族碳環。除非另外規定，否則碳環一般為3員至8員環。舉例而言，完全飽和之環烷基包括諸如環丙基、環丁基、環戊基、環己基及其類似基團之基團。 The term "cycloalkyl" refers to a non-aromatic carbocyclic ring that is fully hydrogenated and is present in a single ring form. Unless otherwise specified, carbocycles are typically from 3 to 8 membered rings. For example, fully saturated cycloalkyl groups include groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

術語「雜芳基」係指含有5至14個環原子之芳環結構，其中至少一個環原子為雜原子(亦即，氧、氮或硫)，剩餘環原子係獨立地選自由碳、氧、氮及硫組成之群。雜芳基可為單環或2或3稠環。雜芳基取代基之實例包括6員環取代基，諸如吡啶基、吡嗪基、嘧啶基及嗒嗪基；5員環取代基，諸如***基、咪唑基、呋喃基、噻吩基、吡唑基、噁唑基、異噁唑基、噻唑基、1,2,3-、1,2,4-、1,2,5-或1,3,4-噁二唑基及異噻唑基；6/5員稠環取代基，諸如苯并硫呋喃基、異苯并硫呋喃基、苯并異噁唑基、苯并噁唑基、嘌呤基及苯鄰甲內醯胺基；及6/6員稠環，諸如喹啉基、異喹啉基、啉基、喹唑啉基及1,4-苯并噁嗪基。在具有雜芳基取代基之基團中，與該基團結合之雜芳基取代基之環原子可為至少一個雜原子，或其可為環碳原子，其中環碳原子可與至少一個雜原子在同一個環中，或其中環碳原子可與至少一個雜原子在不同環中。類似地，若雜芳基取代基又經基團或取代基取代，則該基團或取代基可與至少一個雜原子結合，或可與環碳原子結合，其中環碳原子可與至少一個雜原子在同一環中或其中環碳原子可與至少一個雜原子在不同環中。術語「雜芳基」亦包括吡啶基N-氧化物及含有吡啶N-氧化物環之基團。 The term "heteroaryl" refers to an aromatic ring structure containing from 5 to 14 ring atoms, wherein at least one ring atom is a hetero atom (ie, oxygen, nitrogen or sulfur) and the remaining ring atoms are independently selected from carbon and oxygen. , a group of nitrogen and sulfur. The heteroaryl group can be a single ring or a 2 or 3 fused ring. Examples of heteroaryl substituents include 6-membered ring substituents such as pyridyl, pyrazinyl, pyrimidinyl and pyridazinyl; 5-membered ring substituents such as triazolyl, imidazolyl, furyl, thienyl, pyridyl Azyl, oxazolyl, isoxazolyl, thiazolyl, 1,2,3-, 1,2,4-, 1,2,5- or 1,3,4-oxadiazolyl and isothiazolyl 6/5 member fused ring substituents such as benzothiofuranyl, isobenzothiofuranyl, benzoisoxazolyl, benzoxazolyl, fluorenyl and benzopyridinium; and 6 /6 member fused ring, such as quinolyl, isoquinolinyl, A phenyl group, a quinazolinyl group, and a 1,4-benzoxazinyl group. In the group having a heteroaryl substituent, the ring atom of the heteroaryl substituent bonded to the group may be at least one hetero atom, or it may be a ring carbon atom, wherein the ring carbon atom may be at least one hetero Atoms are in the same ring, or wherein a ring carbon atom can be in a different ring than at least one heteroatom. Similarly, if a heteroaryl substituent is substituted by a group or a substituent, the group or substituent may be bonded to at least one hetero atom or may be bonded to a ring carbon atom, wherein the ring carbon atom may be bonded to at least one Atoms may be in the same ring or in which a ring carbon atom may be in a different ring than at least one heteroatom. The term "heteroaryl" also includes pyridyl N-oxides and groups containing a pyridine N-oxide ring.

單環雜芳基之實例包括呋喃基、二氫呋喃基、四氫呋喃基、噻吩基(亦稱為「硫代呋喃基」)、二氫噻吩基、四氫噻吩基、吡咯基、 異吡咯基、吡咯啉基、吡咯啶基、咪唑基、異咪唑基、咪唑啉基、咪唑啶基、吡唑基、吡唑啉基、吡唑啶基、***基、四唑基、二硫醇基、氧硫唑基、噁唑基、異噁唑基、噻唑基、異噻唑基、噻唑啉基、異噻唑啉基、噻唑啶基、異噻唑啶基、噻二唑基、噁噻唑基、噁二唑基(包括噁二唑基、1,2,4-噁二唑基(亦稱為「唑肟基(azoximyl)」)、1,2,5-噁二唑基(亦稱為「呋呫基」)或1,3,4-噁二唑基)、噁***基(包括1,2,3,4-噁***基或1,2,3,5-噁***基)、二噁唑基(包括1,2,3-二噁唑基、1,2,4-二噁唑基、1,3,2-二噁唑基或1,3,4-二噁唑基)、噁噻唑基、氧硫唑基、氧硫基、哌喃基(包括1,2-哌喃基或1,4-哌喃基)、二氫哌喃基、吡啶基(亦稱為「嗪基」)、哌啶基、二嗪基(包括嗒嗪基(亦稱為「1,2-二嗪基」)、嘧啶基(pyrimidinyl)(亦稱為「1,3-二嗪基」或「嘧啶基(pyrimidyl)」)或吡嗪基(亦稱為「1,4-二嗪基」))、哌嗪基、三嗪基(包括s-三嗪基(亦稱為「1,3,5-三嗪基」)、as-三嗪基(亦稱為1,2,4-三嗪基)及v-三嗪基(亦稱為「1,2,3-三嗪基」))、噁嗪基(包括1,2,3-噁嗪基、1,3,2-噁嗪基、1,3,6-噁嗪基(亦稱為「戊噁唑基」)、1,2,6-噁嗪基或1,4-噁嗪基)、異噁嗪基(包括鄰異噁嗪基或對異噁嗪基)、噁唑啶基、異噁唑啶基、噁噻嗪基(包括1,2,5-噁噻嗪基或1,2,6-噁噻嗪基)、噁二嗪基(包括1,4,2-噁二嗪基或1,3,5,2-噁二嗪基)、嗎啉基、氮呯基、氧呯基、二苯并硫雜七環基(thiepinyl)及二氮呯基。 Examples of monocyclic heteroaryl groups include furyl, dihydrofuranyl, tetrahydrofuranyl, thienyl (also known as "thiofuranyl"), dihydrothienyl, tetrahydrothiophenyl, pyrrolyl, isopyrrolyl, Pyrrolinyl, pyrrolidinyl, imidazolyl, isoimidazolyl, imidazolinyl, imidazolidinyl, pyrazolyl, pyrazolinyl, pyrazolyl, triazolyl, tetrazolyl, dithiol, Oxazozolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, thiazolinyl, isothiazolinyl, thiazolidinyl, isothiazolidinyl, thiadiazolyl, oxathiazolyl, cacao Azolyl (including oxadiazolyl, 1,2,4-oxadiazolyl (also known as "azoximyl"), 1,2,5-oxadiazolyl (also known as "furazan"") or 1,3,4-oxadiazolyl), oxatriazole (including 1,2,3,4-oxatriazolyl or 1,2,3,5-oxatriazole), Oxazolyl (including 1,2,3-bisoxazolyl, 1,2,4-bisoxazolyl, 1,3,2-bisoxazolyl or 1,3,4-bisoxazolyl), Oxythiazolyl, oxazolidine, oxysulfide , piperidyl (including 1,2-piperidyl or 1,4-piperidyl), dihydropentanyl, pyridyl (also known as "azinyl"), piperidinyl, diazinyl ( Including pyridazinyl (also known as "1,2-diazino"), pyrimidinyl (also known as "1,3-diazino" or "pyrimidyl") or pyrazinyl (also known as "1,4-diazinyl")), piperazinyl, triazinyl (including s-triazinyl (also known as "1,3,5-triazinyl"), as-three Azinyl (also known as 1,2,4-triazinyl) and v-triazinyl (also known as "1,2,3-triazinyl")),oxazinyl (including 1,2,3) -oxazinyl, 1,3,2-oxazinyl, 1,3,6-oxazinyl (also known as "pentazozolyl"), 1,2,6-oxazinyl or 1,4- Oxazinyl), isoxazinyl (including o-isooxazinyl or p-oxazinyl), oxazolidinyl, isoxazolidinyl, oxathiazinyl (including 1,2,5-oxazine) Or 1,2,6-oxathiazinyl), oxadiazinyl (including 1,4,2-oxadiazinyl or 1,3,5,2-oxadiazinyl), morpholinyl, nitrogen Sulfhydryl, oxonyl, dibenzothiazepine (thiepinyl) and diazinyl.

2-稠環雜芳基之實例包括吲嗪基、吡啶基、哌喃并吡咯基、4H-喹嗪基、嘌呤基、啶基、吡啶并吡啶基(包括吡啶并[3,4-b]-吡啶基、吡啶并[3,2-b]-吡啶基或吡啶并[4,3-b]-吡啶基)及喋啶基、吲哚基、異吲哚基、假吲哚基(indoleninyl)、異吲唑基、苯并吖嗪基(benzazinyl)、呔嗪基、喹喏啉基、喹唑啉基、苯并二嗪基、苯并哌喃基、苯并硫哌喃基、苯并噁唑基、吲哚噁嗪基、苯鄰甲內醯胺基、苯并間二氧雜環戊烯基、苯并二氧雜環己烷基、苯并噁二唑基、苯并 呋喃基、異苯并呋喃基、苯并噻吩基、異苯并噻吩基、苯并噻唑基、苯并噻二唑基、苯并咪唑基、苯并***基、苯并噁嗪基、苯并異噁嗪基及四氫異喹啉基。 Examples of the 2-fused ring heteroaryl group include pyridazinyl, pyridyl, piperacyryryl, 4H-quinazinyl, fluorenyl, Pyridyl, pyridopyridyl (including pyrido[3,4-b]-pyridyl, pyrido[3,2-b]-pyridyl or pyrido[4,3-b]-pyridyl) and anthracene Pyridyl, fluorenyl, isodecyl, indoleninyl, isoxazolyl, benzazinyl, pyridazinyl, quinoxalinyl, quinazolinyl, benzo Diazido, benzopipetanyl, benzothiopyranyl, benzoxazolyl, oxazinyl, benzo-indolylamine, benzodioxolyl, benzo Dioxanyl, benzooxadiazolyl, benzofuranyl, isobenzofuranyl, benzothienyl, isobenzothiophenyl, benzothiazolyl, benzothiadiazolyl, benzene And imidazolyl, benzotriazolyl, benzoxazinyl, benzisooxazinyl and tetrahydroisoquinolinyl.

3-稠環雜芳基或雜環烷基之實例包括5,6-二氫-4H-咪唑并[4,5,1-ij]喹啉、4,5-二氫咪唑并[4,5,1-hi]吲哚、4,5,6,7-四氫咪唑并[4,5,1-jk][1]苯并氮呯及二苯并呋喃基。 Examples of 3-fused ring heteroaryl or heterocycloalkyl include 5,6-dihydro-4H-imidazo[4,5,1-ij]quinoline, 4,5-dihydroimidazo[4,5 , 1-hi] 吲哚, 4,5,6,7-tetrahydroimidazo[4,5,1-jk][1]benzazepine and dibenzofuranyl.

稠環雜芳基之其他實例包括苯并-稠合雜芳基，諸如吲哚基、異吲哚基(isoindolyl)(亦稱為「異吲哚基(isobenzazolyl)」或「假異吲哚基」)、假吲哚基(indoleninyl)(亦稱為「假吲哚基(pseudoindolyl)」)、異吲唑基(亦稱為「苯并吡唑基」)、苯并吖嗪基(包括喹啉基(亦稱為「1-苯并吖嗪基」)或異喹啉基(亦稱為「2-苯并吖嗪基」))、呔嗪基、喹喏啉基、喹唑啉基、苯并二嗪基(包括啉基(亦稱為「1,2-苯并二嗪基」)或喹唑啉基(亦稱為「1,3-苯并二嗪基」))、苯并哌喃基(包括「基」或「異基」)、苯并硫哌喃基(亦稱為「硫基」)、苯并噁唑基、吲嗪基(indoxazinyl)(亦稱為「苯并異噁唑基」)、苯鄰甲內醯胺基、苯并間二氧雜環戊烯基、苯并二氧雜環己烷基、苯并噁二唑基、苯并呋喃基(亦稱為「薰草酮基(coumaronyl)」)、異苯并呋喃基、苯并噻吩基(benzothienyl)(亦稱為「苯并噻吩基(benzothiophenyl)」、「硫萘次甲基」或「苯并硫呋喃基」)、異苯并噻吩基(亦稱為「異苯并噻吩基」、「異硫萘次甲基」或「異苯并硫呋喃基」)、苯并噻唑基、苯并噻二唑基、苯并咪唑基、苯并***基、苯并噁嗪基(包括1,3,2-苯并噁嗪基、1,4,2-苯并噁嗪基、2,3,1-苯并噁嗪基或3,1,4-苯并噁嗪基)、苯并異噁嗪基(包括1,2-苯并異噁嗪基或1,4-苯并異噁嗪基)、四氫異喹啉基、咔唑基、基及吖啶基。 Other examples of fused ring heteroaryl groups include benzo-fused heteroaryl groups such as fluorenyl, isoindolyl (also known as "isobenzazolyl" or "false isodecyl""), indoleninyl (also known as "pseudoindolyl"), isoxazolyl (also known as "benzopyrazolyl"), benzoxazinyl (including quinolin) Lolinyl (also known as "1-benzoxazinyl") or isoquinolinyl (also known as "2-benzoxazinyl"), pyridazinyl, quinoxalinyl, quinazolinyl Benzodiazinyl (including Lolinyl (also known as "1,2-benzodiozinyl") or quinazolinyl (also known as "1,3-benzodiazinyl")), benzopyranyl (including " Base or different Benzo), benzothiopyranyl (also known as "sulfur ")", benzoxazolyl, indoxazinyl (also known as "benzoxazole"), benzopyridinium, benzodioxolane, benzene And dioxanyl, benzooxadiazolyl, benzofuranyl (also known as "coumaronyl"), isobenzofuranyl, benzothienyl (also Known as "benzothiophenyl", "thionaphthalenyl" or "benzothiofuranyl", isobenzothiophenyl (also known as "isobenzothiophenyl", "isothionaphthalene"Hypomethyl" or "isobenzothiofuranyl"), benzothiazolyl, benzothiadiazolyl, benzimidazolyl, benzotriazolyl, benzoxazinyl (including 1,3,2 - benzoxazinyl, 1,4,2-benzoxazinyl, 2,3,1-benzoxazinyl or 3,1,4-benzoxazinyl), benzoisooxazinyl (including 1,2-benzisoxazinyl or 1,4-benzoxazinyl), tetrahydroisoquinolinyl, carbazolyl, Base and acridinyl.

術語「雜環」係指含有總計3至14個環原子之飽和或部分飽和環結構。至少一個環原子為雜原子(亦即氧、氮或硫)，其餘環原子係獨 立地選自由碳、氧、氮及硫組成之群。或者，雜環可包含2或3個稠合在一起之環，其中至少一個該環含有雜原子作為環原子(例如，氮、氧或硫)。在具有雜環烷基取代基之基團中，與該基團結合之雜環取代基之環原子可為至少一個雜原子，或可為環碳原子，其中環碳原子可與至少一個雜原子在同一個環中，或其中環碳原子可與至少一個雜原子在不同環中。類似地，若雜環取代基又經基團或取代基取代，則該基團或取代基可與至少一個雜原子結合，或可與環碳原子結合，其中環碳原子可與至少一個雜原子在同一環中或其中環碳原子可與至少一個雜原子在不同環中。 The term "heterocycle" refers to a saturated or partially saturated ring structure containing a total of from 3 to 14 ring atoms. At least one ring atom is a hetero atom (ie, oxygen, nitrogen or sulfur), and the remaining ring atoms are independent The site is selected from the group consisting of carbon, oxygen, nitrogen and sulfur. Alternatively, the heterocyclic ring may contain 2 or 3 rings fused together, at least one of which contains a hetero atom as a ring atom (for example, nitrogen, oxygen or sulfur). In the group having a heterocyclic alkyl substituent, the ring atom of the heterocyclic substituent bonded to the group may be at least one hetero atom, or may be a ring carbon atom, wherein the ring carbon atom may be bonded to at least one hetero atom In the same ring, or wherein the ring carbon atom may be in a different ring than the at least one hetero atom. Similarly, if a heterocyclic substituent is substituted by a group or a substituent, the group or substituent may be bonded to at least one hetero atom or may be bonded to a ring carbon atom, wherein the ring carbon atom may be bonded to at least one hetero atom. The ring carbon atoms may be in a different ring from the at least one heteroatom in the same ring or in the ring.

併入本說明書中且構成說明書之一部分的附圖說明實施例且與【實施方式】一起說明所揭示之化合物、組合物、方法及用途。 BRIEF DESCRIPTION OF THE DRAWINGS The accompanying drawings, which are incorporated in the specification of the claims

圖1展示對本發明之代表性化合物在各種劑量下之活體內小鼠功效研究。 Figure 1 shows an in vivo mouse efficacy study of representative compounds of the invention at various doses.

圖2展示本發明之代表性化合物抑制病毒RNA合成。 Figure 2 shows a representative compound of the invention inhibiting viral RNA synthesis.

圖3展示DENV-1 NS4B中之I64V取代對本發明之代表性化合物之活性的作用。 Figure 3 shows the effect of I64V substitution in DENV-1 NS4B on the activity of representative compounds of the invention.

本發明提供適用於治療由病毒感染(特定而言為由登革熱病毒引起之病毒感染)引起之疾病的化合物及其醫藥組合物。以下列舉之實施例包括於本發明中。 The present invention provides compounds and pharmaceutical compositions thereof suitable for use in the treatment of diseases caused by viral infections, in particular viral infections caused by dengue viruses. The examples listed below are included in the present invention.

1.一種式(I)之化合物： 1. A compound of formula (I):

其中R1為F、Cl、Br、CF3、OCH3、(C1-C6)烷基或環烷基；R2為H、F、Cl、Br、(C1-C6)烷基或環烷基；或R1與R2連同其所連接之芳族碳原子一起形成稠合1,3-間二氧雜環戊烯基； R3為H、(C1-C6)烷基或環烷基；R4為H、(C1-C6)烷基、環烷基或-C(=O)-NH2；R5為H、(C1-C6)烷基、環烷基或苄基；R6為H、(C1-C6)烷基或環烷基；R7為H、F、Cl或OCH3；R8為F、Cl、Br、CF3、-OCHF2、OCF3、OCH3或(C1-C6)烷基；X1為CH或N；X2為CH或N；X3為CH或N；X4為=O或H2；R1、R2、R3、R4、R5、R6、R7及R8各自視情況獨立地經一或多個獨立的R300取代基取代；R300係選自由H、(C1-C6)烷基、環烷基、羥基、胺基、F、Cl、Br、苯基、雜環及雜芳基組成之群；或其醫藥學上可接受之鹽。 Wherein R 1 is F, Cl, Br, CF 3 , OCH 3 , (C 1 -C 6 )alkyl or cycloalkyl; and R 2 is H, F, Cl, Br, (C 1 -C 6 )alkyl Or a cycloalkyl group; or R 1 and R 2 together with the aromatic carbon atom to which they are attached form a fused 1,3-dioxolyl; R 3 is H, (C 1 -C 6 ) alkane Or a cycloalkyl group; R 4 is H, (C 1 -C 6 )alkyl, cycloalkyl or -C(=O)-NH 2 ; R 5 is H, (C 1 -C 6 )alkyl, a cycloalkyl or benzyl group; R 6 is H, (C 1 -C 6 )alkyl or cycloalkyl; R 7 is H, F, Cl or OCH 3 ; R 8 is F, Cl, Br, CF 3 , -OCHF 2 , OCF 3 , OCH 3 or (C 1 -C 6 )alkyl; X 1 is CH or N; X 2 is CH or N; X 3 is CH or N; X 4 is =O or H 2 ; R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 and R 8 are each optionally substituted by one or more independent R 300 substituents; R 300 is selected from H, (C) a group consisting of 1 -C 6 )alkyl, cycloalkyl, hydroxy, amine, F, Cl, Br, phenyl, heterocyclic and heteroaryl; or a pharmaceutically acceptable salt thereof.

2.實施例1之化合物或其醫藥學上可接受之鹽，其中R1為Cl。 2. The embodiment of the compound or pharmaceutically Example of a pharmaceutically salt thereof, wherein R 1 is Cl.

3.實施例1或實施例2之化合物或其醫藥學上可接受之鹽，其中R2為Cl或F。 3. The compound of Embodiment 1 or Embodiment 2, or a pharmaceutically acceptable salt thereof, wherein R 2 is Cl or F.

4.前述實施例中任一者之化合物或其醫藥學上可接受之鹽，其中R3為H或環丙基。 The compound of any of the preceding embodiments, wherein R 3 is H or cyclopropyl, or a pharmaceutically acceptable salt thereof.

5.前述實施例中任一者之化合物或其醫藥學上可接受之鹽，其中R4為H。 The compound of any of the preceding embodiments, wherein R 4 is H, or a pharmaceutically acceptable salt thereof.

6.前述實施例中任一者之化合物或其醫藥學上可接受之鹽，其中R5為H、甲基或異丙基。 The compound of any of the preceding embodiments, wherein R 5 is H, methyl or isopropyl, or a pharmaceutically acceptable salt thereof.

7.前述實施例中任一者之化合物或其醫藥學上可接受之鹽，其中R6為H或甲基。 The compound of any of the preceding embodiments, wherein R 6 is H or methyl, or a pharmaceutically acceptable salt thereof.

8.前述實施例中任一者之化合物或其醫藥學上可接受之鹽，其中R7為H或Cl。 The compound of any one of the preceding embodiments, wherein R 7 is H or Cl, or a pharmaceutically acceptable salt thereof.

9.前述實施例中任一者之化合物或其醫藥學上可接受之鹽，其中R8為Cl、甲基、異丙基或第三丁基。 The compound of any of the preceding embodiments, wherein R 8 is Cl, methyl, isopropyl or tert-butyl, or a pharmaceutically acceptable salt thereof.

10.前述實施例中任一者之化合物或其醫藥學上可接受之鹽，其中X1為CH。 The compound of any of the preceding embodiments, or a pharmaceutically acceptable salt thereof, wherein X 1 is CH.

11.前述實施例中任一者之化合物或其醫藥學上可接受之鹽，其中X2為N。 The compound of any one of the preceding embodiments, or a pharmaceutically acceptable salt thereof, wherein X 2 is N.

12.前述實施例中任一者之化合物或其醫藥學上可接受之鹽，其中X3為CH。 12. A compound of the acceptable one, or a pharmaceutically acceptable salt of any of embodiment, wherein X 3 is CH.

13.前述實施例中任一者之化合物或其醫藥學上可接受之鹽，其中X4為=O。 13. The foregoing embodiments of a compound according to any one pharmaceutically or the pharmaceutically salt thereof, wherein X 4 is = O.

14.前述實施例中任一者之化合物或其醫藥學上可接受之鹽，其係選自由以下組成之群： 14. A compound of any of the preceding embodiments, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

15.前述實施例中任一者之化合物或其醫藥學上可接受之鹽，其係選自由以下組成之群： 15. A compound of any of the preceding embodiments, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

16.前述實施例中任一者之化合物或其醫藥學上可接受之鹽，其係選自由以下組成之群： 16. A compound of any of the preceding embodiments, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

17.前述實施例中任一者之化合物或其醫藥學上可接受之鹽，其係(R)-5'-氯-3-((S)-1-(4-氯苯基)乙基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮 17. A compound of any of the preceding embodiments, or a pharmaceutically acceptable salt thereof, which is (R)-5'-chloro-3-((S)-1-(4-chlorophenyl)ethyl -4,6-Dihydrospiro[[1,2,3]triazolo[4,5-b]pyridine-7,3'-carboline]-2',5(3H)-dione

18.一種醫藥組合物，其包含治療有效量之如請求項1至17中任一項之化合物或其醫藥學上可接受之鹽及醫藥學上可接受之載劑或賦形劑。 A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

19.實施例18之醫藥組合物，其進一步包含至少一種其他醫藥劑。 19. The pharmaceutical composition of embodiment 18, further comprising at least one other pharmaceutical agent.

20.實施例18或實施例19之醫藥組合物，其中該至少一種其他醫藥劑為西戈斯韋。 20. The pharmaceutical composition of embodiment 18 or embodiment 19, wherein the at least one other pharmaceutical agent is celgosivir.

21.一種治療由病毒感染引起之疾病的方法，其包含向有此需要之個體投與治療有效量之實施例1至17中任一者之化合物或其醫藥學上可接受之鹽的步驟。 21. A method of treating a disease caused by a viral infection comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of any of embodiments 1 to 17 or a pharmaceutically acceptable salt thereof.

22.實施例21之方法，其中該個體係人類。 22. The method of embodiment 21 wherein the system is human.

23.實施例21或實施例22之方法，其中該病毒感染係由登革熱病毒引起。 23. The method of embodiment 21 or embodiment 22, wherein the viral infection is caused by a dengue virus.

24.實施例1至17中任一者所主張之化合物或其醫藥學上可接受之鹽，其係用作藥物。 24. The compound as claimed in any one of embodiments 1 to 17, or a pharmaceutically acceptable salt thereof, for use as a medicament.

25.實施例1至17中任一者所主張之化合物或其醫藥學上可接受之鹽的用途，其係用於製造用以治療由病毒感染引起之疾病的藥物。 25. Use of a compound as claimed in any one of embodiments 1 to 17, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a disease caused by a viral infection.

26.實施例25之用途，其中該病毒感染係由登革熱病毒引起。 26. The use of embodiment 25, wherein the viral infection is caused by a dengue virus.

27.實施例1至17中任一者所主張之化合物或其醫藥學上可接受之鹽，其係NS4B抑制劑。 27. The compound as claimed in any one of embodiments 1 to 17, or a pharmaceutically acceptable salt thereof, which is an NS4B inhibitor.

本發明之化合物可由包括與熟習此項技術者熟知類似之方法的合成途徑來合成，尤其就本文所含之描述而言。起始物質通常可自諸如Aldrich Chemicals(Milwaukee,Wis.)之商業來源獲得或易於使用熟習此項技術者熟知之方法來製備(例如，藉由一般在Louis F.Fieser及Mary Fieser,Reagents for Organic Synthesis，第1至19卷，Wiley,New York(1967-1999版)或Beilsteins Handbuch der organischen Chemie,4,Aufl.編Springer-Verlag,Berlin，包括增刊(亦可經由Beilstein線上資料庫獲得)中描述之方法來製備)。 The compounds of the present invention can be synthesized by synthetic routes including those well known to those skilled in the art, especially as described herein. The starting materials are typically available from commercial sources such as Aldrich Chemicals (Milwaukee, Wis.) or are readily prepared by methods well known to those skilled in the art (e.g., by generally in Louis F. Fieser and Mary Fieser, Reagents for Organic). Synthesis, Volumes 1 to 19, Wiley, New York (1967-1999 edition) or Beilsteins Handbuch der organischen Chemie, 4, Aufl. Edited by Springer-Verlag, Berlin, including supplements (also available through the Beilstein online database) Method to prepare).

為說明目的，下文所述之反應流程提供用於合成本發明之化合物以及重要中間物之可能途徑。關於個別反應步驟之更詳細描述，參見下文實例章節。熟習此項技術者將瞭解，其他合成途徑可用於合成本發明之化合物。儘管流程中描述且下文論述特定起始物質及試劑，但其他起始物質及試劑可易於替代以提供多種衍生物及/或反應條件。另外，藉由下文所述方法製備之許多化合物可使用熟習此項技術者熟知之習知化學方法根據本發明來進一步改質。 For illustrative purposes, the reaction schemes described below provide a possible route for the synthesis of the compounds of the invention as well as important intermediates. For a more detailed description of the individual reaction steps, see the Examples section below. Those skilled in the art will appreciate that other synthetic routes can be used to synthesize the compounds of the present invention. Although specific starting materials and reagents are described in the schemes and discussed below, other starting materials and reagents can be readily substituted to provide a variety of derivatives and/or reaction conditions. In addition, many of the compounds prepared by the methods described below can be further modified in accordance with the present invention using conventional chemical methods well known to those skilled in the art.

在本發明化合物之製備中，可有必要保護中間物之遠端官能基。對此保護之需要將視遠端官能基之性質及製備方法之條件而變化。對此保護之需要易於由熟習此項技術者來確定。關於保護基及其使用之一般性描述，參見T.W.Greene,Protective Groups in Organic Synthesis,John Wiley & Sons,New York,1991。 In the preparation of the compounds of the invention, it may be necessary to protect the distal functional groups of the intermediate. The need for this protection will vary depending on the nature of the remote functional group and the conditions of the preparation process. The need for this protection is readily determined by those skilled in the art. For a general description of protecting groups and their use, see T. W. Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991.

流程1至3(下文)描述用於生產式(I)化合物之可能途徑。可藉由使用實質上光學純之起始物質或藉由分離層析、再結晶或此項技術中熟知之其他分離技術來製備實質上光學純之式(I)化合物。關於更詳細描述，參見下文實例章節。 Schemes 1 to 3 (below) describe possible routes for the production of compounds of formula (I). Substantially optically pure compounds of formula (I) can be prepared by using substantially optically pure starting materials or by separation chromatography, recrystallization, or other separation techniques well known in the art. For a more detailed description, see the example section below.

在流程1下，使經由下文流程2中所示之各種途徑製得之胺基酯1在鹼性條件下水解以提供相應酸2。酸2、經取代之靛紅3及麥氏酸(Meldrum's acid)衍生物4在乙酸存在下之三組分縮合提供縮合產物5，將其藉由對掌性HPLC進一步純化以產生R-螺-對映異構體產物6。 Under Scheme 1, the aminoester 1 prepared via the various routes shown in Scheme 2 below is hydrolyzed under basic conditions to provide the corresponding acid 2 . The three-component condensation of acid 2 , substituted eosin 3 and Meldrum's acid derivative 4 in the presence of acetic acid provides a condensation product 5 which is further purified by palmitic HPLC to produce R -spiro- Enantiomer product 6 .

流程2展示藉由四種合成途徑來製備胺基酯1。在途徑I)下，使起 始物質7轉化為肼8，其隨後經歷與(E)-2-氰基-3-乙氧基丙烯酸乙酯9縮合反應以提供胺基-羧酸酯10。在途徑II)下，使起始物質11轉化為醇12，隨後醇12與二氮烯13進行光延反應(Mitsunobu reaction)提供中間物14，隨後將其在HCl存在下去保護以提供肼15。肼15與9縮合以產生化合物16。在途徑III)下，使起始物質17轉化為疊氮化物18，隨後經歷與2-氰基乙酸乙酯縮合反應以提供胺基-***19。在途徑IV)下，使酮20還原為醇21，將其轉化為中間物22。22隨後經轉化為肼23，其隨後經歷與9縮合反應以產生24。使胺24封端提供化合物25。 Scheme 2 shows the preparation of the aminoester 1 by four synthetic routes. The starting material 7 is converted to the indole 8 under route I), which is subsequently subjected to a condensation reaction with ( E )-2-cyano-3-ethoxyethyl acrylate 9 to provide the amino-carboxylate 10 . Under route II), starting material 11 is converted to alcohol 12 , followed by a Mitsunobu reaction of alcohol 12 with diazene 13 to provide intermediate 14 , which is then protected in the presence of HCl to provide hydrazine 15 . Condensation of hydrazine 15 and 9 to give compound 16 . The starting material 17 is converted to azide 18 under route III) and subsequently subjected to a condensation reaction with ethyl 2-cyanoacetate to provide the amine-triazole 19 . Under route IV), the ketone 20 is reduced to the alcohol 21 which is converted to the intermediate 22 . 22 is then converted to 肼23 , which then undergoes a condensation reaction with 9 to produce 24 . The amine 24 is capped to provide compound 25 .

在流程3下，在BH3 ．Me2S存在下還原吲哚酮5(如根據流程1所製備)提供化合物26，隨後將其藉由對掌性HPLC純化以產生所需R-異構體27。 Under Process 3, at BH 3 . Me 2 S in the presence of reducing indolinone 5 (e.g., prepared according to Scheme 1) to provide compound 26, which was then purified by chiral HPLC, to yield the desired R- isomer 27.

化合物及中間物可經分離且以化合物本身或其鹽之形式使用。如本文所用，術語「鹽」係指本發明之化合物或中間物之酸加成或鹼加成鹽。「鹽」特定而言包括「醫藥學上可接受之鹽」。術語「醫藥學 上可接受之鹽」係指保留本發明化合物之生物有效性及特性且通常在生物學或其他方面合乎需要之鹽。 The compound and the intermediate can be isolated and used in the form of the compound itself or a salt thereof. As used herein, the term "salt" refers to an acid or base addition salt of a compound or intermediate of the invention. "Salt" specifically includes "pharmaceutically acceptable salts". The term "medicine An "acceptable salt" refers to a salt that retains the biological effectiveness and properties of the compounds of the invention and is generally biologically or otherwise desirable.

醫藥學上可接受之酸加成鹽可由無機酸及有機酸來形成，例如乙酸鹽、天冬胺酸鹽、苯甲酸鹽、苯磺酸鹽、溴化物/氫溴酸鹽、碳酸氫鹽/碳酸鹽、硫酸氫鹽/硫酸鹽、樟腦磺酸鹽、氯化物/鹽酸鹽、氯茶鹼鹽、檸檬酸鹽、乙烷二磺酸鹽、反丁烯二酸鹽、葡庚糖酸鹽、葡糖酸鹽、葡萄糖醛酸鹽、馬尿酸鹽、氫碘酸鹽/碘化物、羥乙磺酸鹽、乳酸鹽、乳二酸鹽、月桂基硫酸鹽、蘋果酸鹽、順丁烯二酸鹽、丙二酸鹽、杏仁酸鹽、甲磺酸鹽、甲基硫酸鹽、萘甲酸鹽、萘磺酸鹽、菸鹼酸鹽、硝酸鹽、十八酸鹽、油酸鹽、草酸鹽、棕櫚酸鹽、雙羥萘酸鹽、磷酸鹽/磷酸氫鹽/磷酸二氫鹽、聚半乳糖醛酸鹽、丙酸鹽、硬脂酸鹽、丁二酸鹽、磺基水楊酸鹽、酒石酸鹽、甲苯磺酸鹽及三氟乙酸鹽。 Pharmaceutically acceptable acid addition salts can be formed from inorganic acids and organic acids, such as acetates, aspartates, benzoates, besylate, bromide/hydrobromide, bicarbonate /carbonate, hydrogen sulfate/sulfate, camphor sulfonate, chloride/hydrochloride, chlorophylline, citrate, ethane disulfonate, fumarate, glucoheptonic acid Salt, gluconate, glucuronate, hippurate, hydroiodide/iodide, isethionate, lactate, lactate, lauryl sulfate, malate, butene Diacid salt, malonate, mandelate, methanesulfonate, methyl sulfate, naphthoate, naphthalene sulfonate, nicotinic acid salt, nitrate salt, octadecanoate, oleate, Oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogen phosphate, polygalacturonate, propionate, stearate, succinate, sulfohydrate Salicylate, tartrate, tosylate and trifluoroacetate.

可衍生鹽之無機酸包括例如鹽酸、氫溴酸、硫酸、硝酸、磷酸及其類似酸。 Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like.

可衍生鹽之有機酸包括例如乙酸、丙酸、乙醇酸、草酸、順丁烯二酸、丙二酸、丁二酸、反丁烯二酸、酒石酸、檸檬酸、苯甲酸、杏仁酸、甲烷磺酸、乙烷磺酸、甲苯磺酸、磺基水楊酸及其類似酸。醫藥學上可接受之鹼加成鹽可由無機及有機鹼來形成。 Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methane Sulfonic acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic acid and the like. Pharmaceutically acceptable base addition salts can be formed from inorganic and organic bases.

可衍生鹽之無機鹼包括例如銨鹽及來自週期表之第I行至第XII行之金屬。在某些實施例中，鹽係衍生自鈉、鉀、銨、鈣、鎂、鐵、銀、鋅及銅；特定而言適合之鹽包括銨鹽、鉀鹽、鈉鹽、鈣鹽及鎂鹽。 Inorganic bases from which salts can be derived include, for example, ammonium salts and metals from rows I to XII of the periodic table. In certain embodiments, the salt is derived from sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc, and copper; particularly suitable salts include ammonium, potassium, sodium, calcium, and magnesium salts. .

可衍生鹽之有機鹼包括例如一級胺、二級胺及三級胺；經取代之胺，包括天然產生之經取代之胺；環胺；鹼性離子交換樹脂及其類似物。某些有機胺包括異丙胺、苄星(benzathine)、膽茶鹼 (cholinate)、二乙醇胺、二乙胺、離胺酸、葡甲胺(meglumine)、哌嗪及緩血酸胺(tromethamine)。 Organic bases from which salts can be derived include, for example, primary amines, secondary amines, and tertiary amines; substituted amines, including naturally occurring substituted amines; cyclic amines; basic ion exchange resins and the like. Some organic amines include isopropylamine, benzathine, and catechin (cholinate), diethanolamine, diethylamine, lysine, meglumine, piperazine and tromethamine.

本發明之醫藥學上可接受之鹽可藉由習知化學方法自母體化合物(鹼性或酸性部分)來合成。一般而言，此等鹽可藉由使此等化合物之游離酸形式與化學計量之量的適當鹼(諸如Na、Ca、Mg或K之氫氧化物、碳酸鹽、碳酸氫鹽或其類似物)反應或藉由使此等化合物之游離鹼形式與化學計量之量的適當酸反應來製備。該等反應通常在水中或在有機溶劑中或在二者之混合物中進行。一般而言，若可實行，則需要使用非水性介質，如***、乙酸乙酯、乙醇、異丙醇或乙腈。其他適合鹽之清單可見於例如「Remington's Pharmaceutical Sciences」，第20版，Mack Publishing Company,Easton,Pa.,(1985)；及Stahl與Wermuth之「Handbook of Pharmaceutical Salts：Properties,Selection,and Use」(Wiley-VCH,Weinheim,Germany,2002)中。 The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound (alkaline or acidic moiety) by conventional chemical methods. In general, such salts can be obtained by subjecting the free acid form of such compounds to a stoichiometric amount of a suitable base such as a hydroxide, carbonate, bicarbonate or the like of Na, Ca, Mg or K. The reaction is prepared or by reacting the free base form of such compounds with a stoichiometric amount of the appropriate acid. These reactions are usually carried out in water or in an organic solvent or a mixture of the two. In general, if practicable, a non-aqueous medium such as diethyl ether, ethyl acetate, ethanol, isopropanol or acetonitrile is required. A list of other suitable salts can be found, for example, in "Remington's Pharmaceutical Sciences", 20th Edition, Mack Publishing Company, Easton, Pa., (1985); and "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" by Stahl and Wermuth ( Wiley-VCH, Weinheim, Germany, 2002).

本文中給出之任何式亦意欲表示化合物之未經標記形式以及經同位素標記形式。經同位素標記之化合物具有由本文中給出之式描繪之結構，但一或多個原子經具有選定原子質量或質量數之原子置換。可併入本發明化合物中之同位素之實例包括氫、碳、氮、氧、磷、氟及氯之同位素，諸如分別為2H、3H、11C、13C、14C、15N、18F31P、32P、35S、36Cl、125I。本發明包括如本文所定義之各種經同位素標記之化合物，例如其中存在諸如3H、13C及14C之放射性同位素之彼等化合物。該等經同位素標記之化合物適用於代謝研究(使用14C)；反應動力學研究(例如使用2H或3H)；偵測或成像技術，諸如正電子發射斷層攝影法(PET)或單光子發射電腦斷層攝影法(SPECT)，包括藥物或受質組織分佈檢定；或對患者之放射性治療。特定而言，18F或經標記之化合物可尤其適用於PET或SPECT研究。本發明之經同位素標記之化合物一般可藉由進行流程或下文所述實例及製備中所揭示之程序，藉 由以易於獲得之經同位素標記之試劑取代未經同位素標記之試劑來製備。 Any formula given herein is also intended to indicate unlabeled forms of the compounds as well as isotopically labeled forms. An isotopically labeled compound has a structure depicted by the formula given herein, but one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes which may be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as 2 H, 3 H, 11 C, 13 C, 14 C, 15 N, 18, respectively. F 31 P, 32 P, 35 S, 36 Cl, 125 I. The invention includes various isotopically labeled compounds as defined herein, such as compounds in which radioisotopes such as 3 H, 13 C and 14 C are present. Such isotopically labeled compounds are suitable for metabolic studies (using 14 C); reaction kinetic studies (eg using 2 H or 3 H); detection or imaging techniques such as positron emission tomography (PET) or single photons Launch computed tomography (SPECT), including drug or matrix distribution assays; or radiotherapy for patients. In particular, 18 F or labeled compounds may be particularly useful for PET or SPECT studies. The isotopically-labeled compounds of the present invention can generally be prepared by substituting an isotope-labeled reagent with a readily available isotopically-labeled reagent by procedures or procedures disclosed in the Examples and Preparations described below.

另外，用較重同位素，尤其為氘(亦即2H或D)進行取代可提供由較大代謝穩定性所致之某些治療優勢，例如活體內半衰期增加、劑量需求降低、Cyp450抑制作用(競爭性或時間依賴性)降低或治療指數改良。舉例而言，以氘進行取代可調節未氘化化合物之不良副作用，諸如競爭性之Cyp450抑制作用、時間依賴性Cyp450鈍化等。應瞭解，本文中之氘應視為本發明化合物中之取代基(包括二聚體之單體及連接子部分)。該較重同位素(詳言之為氘)之濃度可由同位素富集因數來定義。如本文所用之術語「同位素富集因素」意謂指定同位素之同位素豐度與天然豐度之間的比率。若本發明化合物中之取代基表示為氘，則該化合物中各指定氘原子之同位素富集因數為至少3500(各指定氘原子上52.5%氘併入)、至少4000(60%氘併入)、至少4500(67.5%氘併入)、至少5000(75%氘併入)、至少5500(82.5%氘併入)、至少6000(90%氘併入)、至少6333.3(95%氘併入)、至少6466.7(97%氘併入)、至少6600(99%氘併入)或至少6633.3(99.5%氘併入)。 Further, substitution with heavier isotopes, particularly deuterium (i.e., 2 H or D) may be substituted for certain therapeutic advantages resulting from greater metabolic stability provided due to the, for example, increased in vivo half-life, reduced dosage requirements, CYP450 inhibition ( Competitive or time dependent) reduction or treatment index improvement. For example, substitution with hydrazine can modulate undesirable side effects of the undeuterated compound, such as competitive Cyp450 inhibition, time-dependent Cyp450 passivation, and the like. It will be understood that the oxime herein is to be considered as a substituent (including a monomer and a linker moiety of the dimer) in the compounds of the present invention. The concentration of the heavier isotope (detailed in 氘) can be defined by the isotope enrichment factor. The term "isotopic enrichment factor" as used herein means the ratio between the isotope abundance of a given isotope and the natural abundance. If the substituent in the compound of the invention is represented by hydrazine, the isotopic enrichment factor for each of the specified ruthenium atoms in the compound is at least 3500 (52.5% 氘 on each designated ruthenium atom), at least 4000 (60% 氘 incorporation) At least 4500 (67.5% 氘 incorporation), at least 5000 (75% 氘 incorporation), at least 5500 (82.5% 氘 incorporation), at least 6000 (90% 氘 incorporation), at least 6333.3 (95% 氘 incorporation) At least 6466.7 (97% 氘 incorporation), at least 6600 (99% 氘 incorporation) or at least 6633.3 (99.5% 氘 incorporation).

本發明之經同位素標記之化合物一般可由熟習此項技術者已知之習知技術或藉由與隨附實例及製備中所述類似之方法，使用適當經同位素標記之試劑代替先前使用之未經標記之試劑來製備。 Isotopically labeled compounds of the present invention can generally be replaced by conventionally known isotopically labeled reagents by conventional techniques known to those skilled in the art or by methods analogous to those described in the accompanying Examples and Preparations. The reagent is prepared.

本發明之醫藥學上可接受之溶劑合物包括其中結晶化溶劑可經同位素(例如，D2O、d6-丙酮、d6-DMSO)取代之彼等溶劑合物。 The pharmaceutically acceptable solvates of the present invention include those in which the crystallization solvent can be substituted with isotopes (e.g., D 2 O, d 6 -acetone, d 6 -DMSO).

熟習此項技術者將意識到，本發明之化合物可含有對掌性中心且因此可存在不同異構形式。如本文所用，術語「異構體」係指具有相同分子式但原子之排列及組態不同之不同化合物。亦如本文所用，術語「光學異構體」或「立體異構體」係指各種立體異構組態中之任一者，其可在本發明之既定化合物中存在且包括幾何異構體。應瞭 解，取代基可連接在碳原子之對掌性中心處。因此，本發明包括化合物之對映異構體、非對映異構體或外消旋體。 Those skilled in the art will recognize that the compounds of the present invention may contain a palmitic center and thus may exist in different isomeric forms. As used herein, the term "isomer" refers to a different compound having the same molecular formula but differing in the arrangement and configuration of the atoms. Also as used herein, the term "optical isomer" or "stereoisomer" refers to any of a variety of stereoisomeric configurations which may be present in the intended compounds of the invention and include geometric isomers. Should The substituent can be attached to the palm center of the carbon atom. Thus, the invention includes enantiomers, diastereomers or racemates of the compounds.

「對映異構體」係一對彼此呈不重迭鏡像之立體異構體。一對對映異構體之1：1混合物為「外消旋」混合物。適當時，該術語用以表示外消旋混合物。「非對映異構體」為具有至少兩個不對稱原子，但其彼此不為鏡像之立體異構體。根據Cahn- lngold- Prelog R-S系統來規定絕對立體化學。當化合物為純對映異構體時，每個對掌性碳處之立體化學可由R或S規定。絕對組態未知之解析化合物可視其在鈉D線之波長下使平面偏振光旋轉之方向(右旋或左旋)而指定為(+)或(-)。或者，可經由對掌性HPLC由相應對映異構體/非對映異構體各自之滯留時間來定義解析化合物。 An "enantiomer" is a pair of stereoisomers that are not overlapping each other. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture. The term is used to denote a racemic mixture, where appropriate. A "diastereomer" is a stereoisomer that has at least two asymmetric atoms but which are not mirror images of each other. Absolute stereochemistry is specified according to the Cahn-lngold-Prelog RS system. When the compound is a pure enantiomer, the stereochemistry of each pair of palmitic carbons can be specified by R or S. The absolute configuration of an unknown analytical compound can be specified as (+) or (-) depending on the direction in which the plane polarized light is rotated (right-handed or left-handed) at the wavelength of the sodium D-line. Alternatively, the analytical compound can be defined by the respective retention times of the respective enantiomers/diastereomers by palmitic HPLC.

本文所述之某些化合物含有一或多個不對稱中心或軸，且因此可產生可根據絕對立體化學定義為(R)-或(S)-之對映異構體、非對映異構體及其他立體異構形式。除非另外規定，否則本發明之化合物意欲包括所有此等可能之異構體，包括外消旋混合物、光學純形式及中間物混合物。光學活性(R)-及(S)-異構體可使用對掌性合成子或對掌性試劑來製備，或使用習知技術來解析(例如，在對掌性SFC或HPLC層析管柱(諸如可由DAICEL公司獲得之CHIRALPAK®及CHIRALCEL®)上，使用適當溶劑或溶劑混合物分離來達成良好分離)。若化合物含有雙鍵，則取代基可為E或Z組態。若化合物含有經二取代之環烷基，則環烷基取代基可具有順式或反式組態。亦意欲包括所有互變異構形式。 Certain compounds described herein contain one or more asymmetric centers or axes and, therefore, may give rise to enantiomers, diastereoisomers of (R)- or (S)-, as defined by absolute stereochemistry. Body and other stereoisomeric forms. Unless otherwise specified, the compounds of the invention are intended to include all such possible isomers, including racemic mixtures, optically pure forms, and intermediate mixtures. The optically active (R)- and (S)-isomers can be prepared using a palmitic synthon or a palmitic reagent, or resolved using conventional techniques (eg, in a palmitic SFC or HPLC chromatography column). (For example, CHIRALPAK® and CHIRALCEL® available from DAICEL), use a suitable solvent or solvent mixture to separate for good separation). If the compound contains a double bond, the substituent can be in an E or Z configuration. If the compound contains a disubstituted cycloalkyl group, the cycloalkyl substituent can have a cis or trans configuration. It is also intended to include all tautomeric forms.

含有能夠充當氫鍵供體及/或受體之基團的本發明之化合物能夠與適合之共晶體形成體形成共晶體。該等共晶體可藉由已知共晶體形成程序由本發明之化合物來製備。該等程序包括碾磨、加熱、共昇華、共熔融或在溶液中使本發明化合物與共晶體形成體在結晶條件下 接觸並分離由此形成之共晶體。適合之共晶體形成體包括WO 2004/078163中所述之彼等。因此，本發明另外提供包含本發明化合物之共晶體。 A compound of the invention containing a group capable of acting as a hydrogen bond donor and/or acceptor is capable of forming a co-crystal with a suitable eutectic former. Such co-crystals can be prepared from the compounds of the invention by known co-crystal formation procedures. Such procedures include milling, heating, co-subliming, co-melting or bringing the compound of the invention and the eutectic formation in solution under crystallization conditions The co-crystal thus formed is contacted and separated. Suitable co-crystal formers include those described in WO 2004/078163. Accordingly, the invention further provides co-crystals comprising a compound of the invention.

本發明之化合物通常以醫藥組合物(例如，本發明之化合物及至少一種醫藥學上可接受之載劑)之形式使用。如本文所用，術語「醫藥學上可接受之載劑」包括普遍認為安全(GRAS)之溶劑、分散介質、界面活性劑、抗氧化劑、防腐劑(例如抗細菌劑、抗真菌劑)、等張劑、鹽、防腐劑、藥物穩定劑、緩衝劑(例如，順丁烯二酸、酒石酸、乳酸、檸檬酸、乙酸、碳酸氫鈉、磷酸鈉及其類似物)及其類似物及其組合，如熟習此項技術者所知(參見例如Remington's Pharmaceutical Sciences，第18版Mack Printing Company,1990，第1289-1329頁)。除了任何習知載劑與活性成份不相容之情形以外，涵蓋其在治療性或醫藥組合物中之用途。為本發明之目的，考慮包含本發明化合物與溶劑(亦即，溶劑合物)或水(亦即，水合物)之醫藥組合物形式之溶劑合物及水合物。 The compounds of the invention are typically employed in the form of a pharmaceutical composition (e.g., a compound of the invention and at least one pharmaceutically acceptable carrier). As used herein, the term "pharmaceutically acceptable carrier" includes solvents, dispersion media, surfactants, antioxidants, preservatives (eg, antibacterial, antifungal agents), isotonics that are generally considered safe (GRAS). Agents, salts, preservatives, drug stabilizers, buffers (for example, maleic acid, tartaric acid, lactic acid, citric acid, acetic acid, sodium hydrogencarbonate, sodium phosphate, and the like), and the like, and combinations thereof, As is known to those skilled in the art (see, for example, Remington's Pharmaceutical Sciences, 18th Edition, Mack Printing Company, 1990, pp. 1289-1329). In addition to any conventional carrier incompatible with the active ingredient, its use in therapeutic or pharmaceutical compositions is contemplated. For the purposes of the present invention, solvates and hydrates in the form of pharmaceutical compositions comprising a compound of the invention and a solvent (i.e., a solvate) or water (i.e., a hydrate) are contemplated.

調配物可使用習知溶解及混合程序來製備。舉例而言，在上文所述一或多種賦形劑存在下，將大塊藥物物質(亦即，本發明之化合物或穩定形式之化合物(例如，與環糊精衍生物或其他已知複合劑之複合物))溶解於適合溶劑中。本發明之化合物通常調配成醫藥劑型以提供可易於控制劑量之藥物及給與患者美觀且可易於處理之產品。 Formulations can be prepared using conventional dissolution and mixing procedures. For example, a bulk drug substance (ie, a compound of the invention or a stable form of a compound (eg, with a cyclodextrin derivative or other known compound) in the presence of one or more excipients as described above The complex of the agent)) is dissolved in a suitable solvent. The compounds of the present invention are typically formulated into pharmaceutical dosage forms to provide a drug which can be easily controlled in dosage and which is aesthetically pleasing and easy to handle.

視用於投與藥物之方法而定，可以多種方式來封裝醫藥組合物(或調配物)以供施用。一般而言，供分配用之物件包括其中存放有適當形式之醫藥調配物之容器。適合容器為熟習此項技術者所熟知且包括諸如瓶(塑料瓶及玻璃瓶)、安瓿、塑料袋、金屬圓筒及其類似物之材料。容器亦可包括防干擾裝配以防止輕易獲取封裝之內含物。另外，容器上附有描述容器內含物之標籤。該標籤亦可包括適當警告。 Depending on the method used to administer the drug, the pharmaceutical composition (or formulation) can be encapsulated for administration in a variety of ways. In general, articles for dispensing include containers in which a suitable form of pharmaceutical formulation is stored. Suitable containers are well known to those skilled in the art and include materials such as bottles (plastic bottles and glass bottles), ampoules, plastic bags, metal cylinders and the like. The container may also include an interference proof assembly to prevent easy access to the contents of the package. In addition, a label describing the contents of the container is attached to the container. The label can also include appropriate warnings.

在某些情況下，有利地與至少一種其他醫藥劑(或治療劑)組合來投與本發明之化合物。本發明之化合物可與一或多種其他治療劑同時、或在其之前或之後投與。或者，本發明之化合物可藉由相同或不同投藥途徑單獨投與，或與其他藥劑於同一醫藥組合物中一起投與。 In certain instances, the compounds of the invention are advantageously administered in combination with at least one other pharmaceutical agent (or therapeutic agent). The compounds of the invention may be administered simultaneously with, or before or after, one or more other therapeutic agents. Alternatively, the compounds of the invention may be administered separately by the same or different routes of administration, or together with other agents in the same pharmaceutical composition.

適合之其他醫藥劑包括(但不限於)干擾素、利巴韋林及利巴韋林類似物、親環素結合劑、HCV NS3蛋白酶抑制劑、HCV NS5a抑制劑、P7抑制劑、進入抑制劑、NS4b抑制劑、α-葡萄糖苷酶抑制劑、宿主蛋白酶抑制劑、免疫調節劑、用於嚴重登革熱之誘發細胞激素或趨化激素之激酶抑制劑、諸如用於血漿滲漏等之症狀緩解劑、表面受體(諸如CLEC5A及DC-SIGN)、核苷及非核苷NS5b抑制劑。 Other pharmaceutical agents suitable for use include, but are not limited to, interferon, ribavirin and ribavirin analogs, cyclophilin binding agents, HCV NS3 protease inhibitors, HCV NS5a inhibitors, P7 inhibitors, entry inhibitors , NS4b inhibitor, α-glucosidase inhibitor, host protease inhibitor, immunomodulator, kinase inhibitor for cytokine or chemokine for severe dengue, such as symptom relief agent for plasma leakage Surface receptors (such as CLEC5A and DC-SIGN), nucleosides and non-nucleoside NS5b inhibitors.

本發明之化合物或其醫藥組合物用於人類通常以治療劑量經口投與。 The compounds of the invention or pharmaceutical compositions thereof are used in humans for oral administration, usually at therapeutic doses.

應瞭解，欲用於治療病毒感染之本發明化合物之劑量範圍視熟習此項技術者已知之因數而定，包括欲治療病狀之宿主、性質及嚴重程度、投藥模式及欲採用之特定物質。 It will be appreciated that the dosage range of the compound of the invention to be used in the treatment of a viral infection will depend on factors known to those skilled in the art, including the host to be treated, the nature and severity, the mode of administration, and the particular substance to be employed.

本發明化合物之每日劑量將隨所採用之化合物、投藥模式、所需之治療及指定之疾病以及醫學領域已知之其他因數(諸如個體之年齡、體重、一般健康狀況、病狀、先前醫藥史及性別及其類似因數)而變化。舉例而言，以每公斤體重約0.1mg至每公斤體重約20mg範圍內，例如每公斤體重約1mg至每公斤體重約10mg範圍內之每日劑量投與本發明之化合物。對於70kg人類而言，通常在以約0.001g至約10g，例如不超過約1公克，例如約0.1g至約0.5g之每日劑量每天至多4次投與本發明之化合物時，可獲得令人滿意之結果。 The daily dosage of a compound of the invention will depend on the compound employed, the mode of administration, the treatment desired and the condition specified, and other factors known in the medical arts (such as the age, weight, general health, condition, prior medical history of the individual) And gender and its similar factors). For example, a compound of the invention is administered at a daily dose ranging from about 0.1 mg per kilogram of body weight to about 20 mg per kilogram of body weight, such as from about 1 mg per kilogram of body weight to about 10 mg per kilogram of body weight. For a 70 kg human, the compound of the invention is typically administered at a daily dose of from about 0.001 g to about 10 g, such as no more than about 1 gram, for example from about 0.1 g to about 0.5 g, up to 4 times a day. The result of satisfaction.

此外，分數次給藥以及交錯給藥可每日或相繼投與，或給藥可為連續輸注或可為快速注射。此外，如治療或預防情形之緊急性所示，本發明化合物之劑量可呈比例性增加或減少。 In addition, fractional administration and staggered administration may be administered daily or sequentially, or administration may be continuous infusion or may be rapid injection. Furthermore, the dosage of the compounds of the invention may be proportionally increased or decreased as indicated by the urgency of the therapeutic or prophylactic situation.

一般而言，化合物、醫藥組合物或其組合之治療有效劑量視個體之物種、體重、年齡及所治療之個別病狀、病症或疾病或其嚴重程度而定。一般熟練之醫師、藥師、臨床醫生或獸醫可容易地確定預防、治療或抑制病症或疾病進程所必需之各活性成份的有效量。 In general, a therapeutically effective amount of a compound, pharmaceutical composition, or combination thereof will depend on the species, weight, age, and individual condition, disorder, or disease or severity of the individual being treated. A generally skilled physician, pharmacist, clinician or veterinarian can readily determine the effective amount of each active ingredient necessary to prevent, treat or inhibit the progression of a condition or disease.

本發明之另一態樣係一種合併製劑形式之包含本發明之化合物及至少一種其他治療劑(或醫藥劑)之產品，其用於同時、單獨或相繼用於治療患有由病毒感染引起之疾病之個體的療法中。 Another aspect of the invention is a product comprising a compound of the invention and at least one other therapeutic agent (or pharmaceutical agent) in the form of a combined preparation for simultaneous, separate or sequential use in the treatment of a disease caused by a viral infection In the treatment of individuals with diseases.

在本發明之組合療法中，本發明之化合物與其他治療劑可由相同或不同製造商來製造及/或調配。此外，本發明之化合物與其他治療劑(或醫藥劑)可一起置於組合療法中：(i)在將組合產品讓與醫師之前(例如，在包含本發明之化合物及其他治療劑或固定劑量組合物之套組的情形下)；(ii)在投藥之前不久由醫師本身(或在醫師指導下)將其置於組合療法中；(iii)在患者本身中，例如在相繼投與本發明之化合物及其他治療劑期間。 In the combination therapies of the invention, the compounds of the invention may be made and/or formulated with other therapeutic agents by the same or different manufacturers. Furthermore, the compounds of the invention may be placed in combination therapy with other therapeutic agents (or pharmaceutical agents): (i) prior to delivery of the combination product to a physician (eg, comprising a compound of the invention and other therapeutic agents or fixed doses) In the case of a kit of compositions); (ii) placed in combination therapy by the physician itself (or under the direction of a physician) shortly before administration; (iii) in the patient itself, for example, in succession of the present invention During the period of compounds and other therapeutic agents.

將非經腸組合物調配成易於投藥且劑量均一之單位劑型係尤其有利的。如本文所用之單位劑型係指適合單位劑量之物理離散單位，每個單位含有經計算與所需醫藥載劑結合產生所需治療作用之預定量的活性成分。該等單位劑型之實例為錠劑(包括加刻痕之錠劑或塗佈錠劑)、膠囊、藥丸、散劑袋、薄片、栓劑、可注射溶液或懸浮液及其類似形式以及其多種隔離形式。 It is especially advantageous to formulate parenteral compositions in unit dosage forms for ease of administration and uniformity. Dosage unit form, as used herein, refers to physically discrete units suitable as unit dosages, each unit containing a predetermined amount of active ingredient calculated to produce the desired therapeutic effect in association with the desired pharmaceutical carrier. Examples of such unit dosage forms are lozenges (including scented lozenges or coated lozenges), capsules, pills, powder sachets, tablets, suppositories, injectable solutions or suspensions, and the like, and various forms thereof .

關於所用其他治療劑之每日劑量將例如視所用化合物、宿主、投藥模式及待治療病狀之嚴重程度而變化。由於可使用之其他治療劑的不同類型，其量可大幅變化，且如上文所述可由常規實驗來確定。 The daily dosage of the other therapeutic agent used will vary, for example, depending on the compound employed, the host, the mode of administration, and the severity of the condition being treated. The amount can vary widely due to the different types of other therapeutic agents that can be used, and can be determined by routine experimentation as described above.

本發明之化合物及至少一種其他治療劑(或醫藥劑)可由任何習知途徑來投與，特定而言經腸(例如以可飲溶液、錠劑或膠囊之形式經口投與)或非經腸(例如以可注射溶液或懸浮液之形式)。 The compound of the present invention and at least one other therapeutic agent (or pharmaceutical agent) can be administered by any conventional means, in particular, enterally (for example, orally in the form of a drinkable solution, lozenge or capsule) or non-menstrual Intestine (for example in the form of an injectable solution or suspension).

以下實例說明本發明之實施例。然而應瞭解，本發明之實施例不限於該等實例之特定詳情，因為根據本發明，一般熟習此項技術者將已知或顯而易見其其他變化形式。 The following examples illustrate embodiments of the invention. However, it should be understood that the embodiments of the invention are not limited to the specific details of the examples, as other variations of the invention will be apparent to those skilled in the art.

在本發明中，參考各種公開案。該等公開案之揭示內容以其全文引用的方式併入本發明中以更全面地描述其所屬之技術水平。 In the present invention, reference is made to various publications. The disclosures of the publications are hereby incorporated by reference in their entirety in their entirety in their entirety in the extent of the extent of the disclosure.

實例Instance

除非另外規定，否則起始物質一般購自非排他性之商業來源，諸如TCI Fine Chemicals(Japan)、Shanghai Chemhere Co.,Ltd.(Shanghai,China)、Aurora Fine Chemicals LLC(San Diego,CA)、FCH Group(Ukraine)、Aldrich Chemicals Co.(Milwaukee,Wis.)、Lancaster Synthesis,Inc.(Windham,N.H.)、Acros Organics(Fairlawn,N.J.)、Maybridge Chemical Company,Ltd.(Cornwall,England)、Tyger Scientific(Princeton,N.J.)、AstraZeneca Pharmaceuticals(London,England)、Chembridge Corporation(USA)、Matrix Scientific(USA)、Conier Chem & Pharm Co.、Ltd(China)、Enamine Ltd(Ukraine)、Combi-Blocks,Inc.(San Diego,USA)、Oakwood Products,Inc.(USA)、Apollo Scientific Ltd.(UK)、Allichem LLC.(USA)及Ukrorgsyntez Ltd(Latvia)。 Unless otherwise specified, starting materials are generally purchased from non-exclusive commercial sources such as TCI Fine Chemicals (Japan), Shanghai Chemhere Co., Ltd. (Shanghai, China), Aurora Fine Chemicals LLC (San Diego, CA), FCH. Group (Ukraine), Aldrich Chemicals Co. (Milwaukee, Wis.), Lancaster Synthesis, Inc. (Windham, NH), Acros Organics (Fairlawn, NJ), Maybridge Chemical Company, Ltd. (Cornwall, England), Tyger Scientific ( Princeton, NJ), AstraZeneca Pharmaceuticals (London, England), Chembridge Corporation (USA), Matrix Scientific (USA), Conier Chem & Pharm Co., Ltd (China), Enamine Ltd (Ukraine), Combi-Blocks, Inc. ( San Diego, USA), Oakwood Products, Inc. (USA), Apollo Scientific Ltd. (UK), Allichem LLC. (USA), and Ukrorgsyntez Ltd (Latvia).

下文中使用之以下縮寫具有相應含義： The following abbreviations used below have corresponding meanings:

實例1Example 1 製備(R)-5-氯-1'-((5-氯吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 Preparation of (R)-5-chloro-1'-((5-chloropyridin-2-yl)methyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[ 3,4-b]pyridine]-2,6'(1'H)-dione

步驟1：製備(5-氯吡啶-2-基)甲醇 Step 1: Preparation of (5-chloropyridin-2-yl)methanol

將5-氯吡啶甲酸甲酯(2g，11.7mmol)於MeOH(20ml)及硼氫化鈉(0.89g，23.3mmol)中之溶液在室溫下攪拌1h。反應完成後，蒸發 溶劑。添加蒸餾水且以二氯甲烷(3×30ml)萃取含水層。將經合併之有機層經鹽水洗滌、經無水Na2SO4乾燥且在減壓下濃縮以提供1.6g(95%產率)呈無水液體狀之(5-氯吡啶-2-基)甲醇。 A solution of methyl 5-chloropicolinate (2 g, 11.7 mmol) in EtOAc (EtOAc) After the reaction was completed, the solvent was evaporated. Distilled water was added and the aqueous layer was extracted with dichloromethane (3×30 mL). The organic layers were washed with brine the combined, dried over anhydrous Na 2 SO 4 dried and concentrated to afford 1.6g (95% yield) of anhydrous liquid (5-chloro-pyridin-2-yl) methanol under reduced pressure.

1H NMR(400MHz,CDCl3)：δ ppm 3.29(br s,1H),4.75(br s,2H),7.25(1H，與CDCl3合併)，7.67(d,J=7.0Hz,1H),8.52(s,1H)。 1 H NMR (400MHz, CDCl 3 ): δ ppm 3.29 (br s, 1H), 4.75 (br s, 2H), 7.25 (1H, merged with CDCl 3), 7.67 (d, J = 7.0Hz, 1H), 8.52 (s, 1H).

LC-MS：m/z 144.0(M+H)+。 LC-MS: m/z 144.0 (M+H) + .

步驟2：製備1-((5-氯吡啶-2-基)甲基)肼-1,2-二甲酸二第三丁酯 Step 2: Preparation of di-t-butyl 1-((5-chloropyridin-2-yl)methyl)indole-1,2-dicarboxylate

在0℃下，向(5-氯吡啶-2-基)甲醇(1.6g，11.2mmol)於THF中之溶液中添加三苯基膦(1.6g，11.188mmol)及肼基甲酸二第三丁酯(4.4g，16.79mmol)且將反應混合物在室溫下攪拌16h。在真空下移除溶劑且藉由使用以20% EtOAc/石油醚(Pet-ether)溶離之100-200矽膠網的管柱層析純化以產生3.5g呈淺黃色液體狀之1-((5-氯吡啶-2-基)甲基)肼-1,2-二甲酸二第三丁酯(76%產率)。 To a solution of (5-chloropyridin-2-yl)methanol (1.6 g, 11.2 mmol) in THF was added triphenylphosphine (1.6 g, 11.188 mmol) and dimethyl carbazate at 0 ° C. Ester (4.4 g, 16.79 mmol) and the mixture was stirred at room temperature for 16 h. The solvent was removed under vacuum and purified by column chromatography using 100-200 EtOAc EtOAc EtOAc EtOAc EtOAc EtOAc EtOAc -Chloropyridin-2-yl)methyl)indole-1,2-dicarboxylic acid di-t-butyl ester (76% yield).

1H NMR(400MHz,CDCl3)：δ ppm 1.46(br s,18H),4.74(br s,2H),7.26(1H，與CDCl3合併)，7.60-7.65(m,1H),8.49(d,J=2.2Hz,1H)。 1 H NMR (400MHz, CDCl 3 ): δ ppm 1.46 (br s, 18H), 4.74 (br s, 2H), 7.26 (1H, merged with CDCl 3), 7.60-7.65 (m, 1H) , 8.49 (d , J = 2.2 Hz, 1H).

LC-MS：m/z 358.1(M+H)+。 LC-MS: m/z 358.1 (M+H) + .

步驟3：製備5-氯-2-(肼基甲基)吡啶鹽酸鹽 Step 3: Preparation of 5-chloro-2-(indolylmethyl)pyridine hydrochloride

在0℃下，向1-((5-氯吡啶-2-基)甲基)肼-1,2-二甲酸二第三丁酯(3.5g，9.8mmol)於二噁烷(20mL)中之溶液中添加二噁烷-HCl(35mL)，繼而添加2滴水。將反應混合物在室溫下攪拌3h直至反應完成，且在真空下移除溶劑並以***濕磨以提供1.3g呈奶白色固體狀之5-氯-2-(肼基甲基)吡啶鹽酸鹽(69%產率)。 To a solution of di-tert-butyl 1-((5-chloropyridin-2-yl)methyl)indole-1,2-dicarboxylate (3.5 g, 9.8 mmol) in dioxane (20 mL) Dioxane-HCl (35 mL) was added to the solution, followed by the addition of 2 drops of water. The reaction mixture was stirred at room temperature for 3 h until the reaction was completed, and the solvent was evaporated and evaporated to dryness with diethyl ether to afford <RTI ID=0.>> Salt (69% yield).

1H NMR(400MHz,DMSO-d6)：δ ppm 4.18(br s,2H),7.53(d,J=8.3Hz,1H),7.95-8.0(m,1H),8.62(d,J=2.2Hz,1H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm 4.18 (br s, 2H), 7.53 (d, J = 8.3Hz, 1H), 7.95-8.0 (m, 1H), 8.62 (d, J = 2.2 Hz, 1H).

LC-MS：m/z 158.0(M+H)+。 LC-MS: m/z 158.0 (M+H) + .

步驟4：製備5-胺基-1-((5-氯吡啶-2-基)甲基)-1H-吡唑-4-甲酸乙酯 Step 4: Preparation of ethyl 5-amino-1-((5-chloropyridin-2-yl)methyl)-1H-pyrazole-4-carboxylate

在室溫下，向5-氯-2-(肼基甲基)吡啶鹽酸鹽(1.3g，8.4mmol)及(E)-2-氰基-3-乙氧基丙烯酸乙酯(1.4g，8.4mmol)於乙酸(6.5ml)中之溶液中添加水(1.3ml)，繼而添加乙酸鈉(1.49g，18.2mmol)，且使所得反應混合物在110℃下維持16h。將反應混合物冷卻至室溫，且在減壓下移除溶劑且藉由使用100-200矽膠網之管柱層析純化。將產物在20% EtOAc/石油醚中溶離以產生800mg呈奶白色固體狀之5-胺基-1-((5-氯吡啶-2-基)甲基)-1H-吡唑-4-甲酸乙酯(57%產率)。 To 5-chloro-2-(indolylmethyl)pyridine hydrochloride (1.3 g, 8.4 mmol) and ( E )-2-cyano-3-ethoxyethyl acrylate (1.4 g) at room temperature Water (1.3 ml) was added to a solution of EtOAc (EtOAc) (EtOAc,EtOAc. The reaction mixture was cooled to room temperature, and the solvent was removed under reduced pressure and purified by column chromatography using a 100-200 silica gel. The product was dissolved in 20% EtOAc / pet ether to afford <RTI ID=0.0> Ethyl ester (57% yield).

1H NMR(400MHz,CDCl3)：δ ppm 1.32(t,J=7.0Hz,3H),4.25 (q,J=7.1Hz,2H),5.18(s,2H),5.73(br s,2H),7.30(d,J=7.9Hz,1H),7.60(s,1H),7.65-7.70(m,1H),8.51(d,J=2.2Hz,1H)。 1 H NMR (400 MHz, CDCl 3 ): δ ppm 1.32 (t, J = 7.0 Hz, 3H), 4.25 (q, J = 7.1 Hz, 2H), 5.18 (s, 2H), 5.73 (br s, 2H) , 7.30 (d, J = 7.9 Hz, 1H), 7.60 (s, 1H), 7.65-7.70 (m, 1H), 8.51 (d, J = 2.2 Hz, 1H).

LC-MS：m/z 281.1(M+H)+。 LC-MS: m/z 281.1 (M+H) + .

步驟5：製備5-胺基-1-((5-氯吡啶-2-基)甲基)-1H-吡唑-4-甲酸鈉 Step 5: Preparation of 5-amino-1-((5-chloropyridin-2-yl)methyl)-1H-pyrazole-4-carboxylate

在室溫下，向5-胺基-1-((5-氯吡啶-2-基)甲基)-1H-吡唑-4-甲酸乙酯(800mg，2.86mmol)於乙醇(8mL)中之溶液中添加水(8mL)，繼而添加氫氧化鈉(228mg，5.7mmol)，且將所得反應混合物在90℃下維持4h。將所得反應混合物冷卻至室溫且濃縮至乾燥以產生5-胺基-1-(3-氯苄基)-1H-吡唑-4-甲酸鈉，其不經進一步純化即用於下一步驟。 Ethyl 5-amino-1-((5-chloropyridin-2-yl)methyl)-1H-pyrazole-4-carboxylate (800 mg, 2.86 mmol) in ethanol (8 mL) Water (8 mL) was added to the solution, followed by sodium hydroxide (228 mg, 5.7 mmol), and the obtained mixture was maintained at 90 ° C for 4 h. The resulting reaction mixture was cooled to EtOAc EtOAc (EtOAc m.

步驟6：製備6-氯-1'-((5-氯吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 Step 6: Preparation of 6-chloro-1 '-((5-chloropyridin-2-yl)methyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3 ,4-b]pyridine]-2,6'(1'H)-dione

在室溫下，向上文之5-胺基-1-(3-氯苄基)-1H-吡唑-4-甲酸鈉於乙酸中之溶液中添加5-氯-靛紅(73mg，0.365mmol)及麥氏酸(52.6mg，0.365mmol)且回流3h。將反應混合物冷卻至室溫且在減壓下移除溶劑。向反應混合物中添加水且在室溫下攪拌15min。將經過濾獲得之固體以水(2×15ml)洗滌且在真空下乾燥。藉由使用二氯甲烷中8%甲醇之溶劑梯度作為溶離劑經矽膠(100-200目)管柱層析純化所得固體以提供200mg呈奶白色固體狀之6-氯-1'-((5-氯吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮(17%產率，對於 兩個步驟而言)。 Add 5-chloro-indigo (73 mg, 0.365 mmol) to a solution of sodium 5-amino-1-(3-chlorobenzyl)-1H-pyrazole-4-carboxylate in acetic acid at room temperature. And McDonald's acid (52.6 mg, 0.365 mmol) and refluxed for 3 h. The reaction mixture was cooled to room temperature and the solvent was removed under reduced pressure. Water was added to the reaction mixture and stirred at room temperature for 15 min. The solid obtained by filtration was washed with water (2×15 mL) and dried under vacuum. The resulting solid was purified by column chromatography on silica gel (100-200 mesh) using solvent gradient of 8% methanol in dichloromethane to afford 200 mg of 6-chloro-1'- (5 -chloropyridin-2-yl)methyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1 'H)-dione (17% yield, for In terms of two steps).

1H NMR(400MHz,DMSO-d6)：δ ppm 2.54(d,J=16.2Hz,1H，與DMSO峰部分重疊)，3.12(d,J=16.2Hz,1H),5.35(q,J=16.2Hz,2H),6.82(s,1H),6.92(d,J=8.3Hz,1H),7.00(d,J=8.3Hz,1H),7.33-7.30(m,2H),7.90(d,J=8.3Hz,1H),8.60(br,s,1H),10.60(br,s,1H),11.10(br,s,1H)。 1 H NMR (400 MHz, DMSO-d 6 ): δ ppm 2.54 (d, J = 16.2 Hz, 1H, partially overlapping with DMSO peak), 3.12 (d, J = 16.2 Hz, 1H), 5.35 (q, J = 16.2 Hz, 2H), 6.82 (s, 1H), 6.92 (d, J = 8.3 Hz, 1H), 7.00 (d, J = 8.3 Hz, 1H), 7.33-7.30 (m, 2H), 7.90 (d, J = 8.3 Hz, 1H), 8.60 (br, s, 1H), 10.60 (br, s, 1H), 11.10 (br, s, 1H).

LC-MS：414.0(M+H)+。 LC-MS: 414.0 (M+H) + .

步驟7：製備(R)-5-氯-1'-((5-氯吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮(實例1) Step 7: Preparation of (R)-5-chloro-1'-((5-chloropyridin-2-yl)methyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyridyl Zoxao[3,4-b]pyridine]-2,6'(1'H)-dione (Example 1)

使500mg 6-氯-1'-((5-氯吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮經受對掌性製備型HPLC(管柱：Chiralpak AS-H(4.6×250mm)5μ；移動相：己烷：EtOH：異丙胺(60：40：0.1)；流速：1.0mL/min；稀釋劑：移動相；UV：258nm)以提供呈白色固體狀之兩種對映異構體：250mg(R)-5-氯-1'-((5-氯吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮(實例1)(峰-1，6.17min於對掌性HPLC中，100% ee)與155mg(S)-5-氯-1'-((5-氯吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮(峰-2，10.48min於對掌性HPLC中，99.1% ee)。實例1之特徵資料展示如下：[α]25.8 D：-99.1(c=0.53、CHCl3)。 500 mg of 6-chloro-1 '-((5-chloropyridin-2-yl)methyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4 -b]pyridine]-2,6'(1'H)-dione was subjected to palmar preparative HPLC (column: Chiralpak AS-H (4.6 x 250 mm) 5μ; mobile phase: hexane: EtOH: isopropylamine (60:40:0.1); flow rate: 1.0 mL/min; diluent: mobile phase; UV: 258 nm) to provide two enantiomers as a white solid: 250 mg (R)-5-chloro-1 '-((5-Chloropyridin-2-yl)methyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2 , 6'(1'H)-dione (Example 1) (peak-1, 6.17 min in palmitic HPLC, 100% ee) and 155 mg ( S )-5-chloro-1'-((5- Chloropyridin-2-yl)methyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1' H)-dione (peak -2, 10.48 min in palmitic HPLC, 99.1% ee). The characteristics of Example 1 are shown below: [α] 25.8 D : -99.1 (c = 0.53, CHCl 3 ).

1H NMR(400MHz,DMSO-d6)：δ ppm 2.54(d,J=16.2Hz,1H，與DMSO峰部分重疊)，3.12(d,J=16.2Hz,1H),5.35(q,J=16.2Hz, 2H),6.82(s,1H),6.92(d,J=8.3Hz,1H),7.00(d,J=8.3Hz,1H),7.33-7.30(m,2H),7.90(d,J=8.3Hz,1H),8.60(br,s,1H),10.60(br,s,1H),11.10(br,s,1H)。 1 H NMR (400 MHz, DMSO-d 6 ): δ ppm 2.54 (d, J = 16.2 Hz, 1H, partially overlapping with DMSO peak), 3.12 (d, J = 16.2 Hz, 1H), 5.35 (q, J = 16.2 Hz, 2H), 6.82 (s, 1H), 6.92 (d, J = 8.3 Hz, 1H), 7.00 (d, J = 8.3 Hz, 1H), 7.33-7.30 (m, 2H), 7.90 (d, J = 8.3 Hz, 1H), 8.60 (br, s, 1H), 10.60 (br, s, 1H), 11.10 (br, s, 1H).

LC-MS：414.0(M+H)+。 LC-MS: 414.0 (M+H) + .

實例2Example 2 製備(R)-5-氯-1'-(2,4-二氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 Preparation of (R)-5-chloro-1'-(2,4-dichlorobenzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4- b]pyridine]-2,6'(1'H)-dione

步驟1：製備(2,4-二氯苄基)肼 Step 1: Preparation of (2,4-dichlorobenzyl) hydrazine

在室溫下，向水合肼(25.2g，503.8mmol)於乙醇(80mL)中之溶液中添加2,4-二氯-1-(氯甲基)苯(10g，50.4mmol)於乙醇(50mL)中之溶液，將所得反應團攪拌14h。反應完成後，在減壓下蒸發溶劑。將殘餘物置於水中之50%氫氧化鈉(50mL)中，以***(3×100mL)萃取水溶液。將經合併之有機層以水(100mL)洗滌、經無水Na2SO4乾燥、過濾且在減壓下濃縮以提供呈無色液體狀之(2,4-二氯苄基)肼(7g，72%產率)。 To a solution of hydrazine hydrate (25.2 g, 503.8 mmol) in ethanol (80 mL) was added 2,4-dichloro-1-(chloromethyl)benzene (10 g, 50.4 mmol) in ethanol (50 mL) The solution was stirred for 14 h. After the reaction was completed, the solvent was evaporated under reduced pressure. The residue was taken up in 50% EtOAc (50 mL)EtOAc. The combined organic layers with water (100 mL), dried over anhydrous Na 2 SO 4, filtered and concentrated to afford a colorless liquid of (2,4-dichlorobenzyl) hydrazine (7g under reduced pressure, 72 %Yield).

1H-NMR(CDCl3)：H-NMR(CDCl3 400MHz,CDCl3)：δ ppm 2.8-3.5(br s,3H),3.99(s,2H),7.3-7.2(m,1H),7.45-7.34(m,2H)。 1 H-NMR (CDCl 3 ): H-NMR (CDCl 3 400 MHz, CDCl 3 ): δ ppm 2.8-3.5 (br s, 3H), 3.99 (s, 2H), 7.3-7.2 (m, 1H), 7.45 -7.34 (m, 2H).

MS：m/z 191.0(M+H)+。 MS: m/z 191.0 (M + H) + .

步驟2：製備5-胺基-1-(2,4-二氯苄基)-1H-吡唑-4-甲酸乙酯 Step 2: Preparation of ethyl 5-amino-1-(2,4-dichlorobenzyl)-1H-pyrazole-4-carboxylate

在室溫下，向(2,4-二氯苄基)肼(7g，36.65mmol)於乙醇(80mL)中之攪拌溶液中添加(E)-2-氰基-3-乙氧基丙烯酸乙酯(6.2g，36.65mmol)。將所得反應混合物在80℃下維持4h。將反應混合物冷卻至室溫、在減壓下移除溶劑。藉由使用石油醚中20-25%乙酸乙酯之溶劑梯度作為溶離劑，經矽膠(100-200目)管柱層析純化粗化合物以提供7g(60.8%產率)呈淺黃色膠狀物之5-胺基-1-(2,4-二氯苄基)-1H-吡唑-4-甲酸乙酯。 Add (E)-2-cyano-3-ethoxy acrylate B to a stirred solution of (2,4-dichlorobenzyl) hydrazine (7 g, 36.65 mmol) in ethanol (80 mL). Ester (6.2 g, 36.65 mmol). The resulting reaction mixture was maintained at 80 ° C for 4 h. The reaction mixture was cooled to room temperature and the solvent was removed under reduced pressure. The crude compound was purified by silica gel (100-200 mesh) column chromatography using a solvent gradient of 20-25% ethyl acetate in petroleum ether as a solvent to afford 7 g (60.8% yield) as a pale yellow gum. 5-Amino-1-(2,4-dichlorobenzyl)-1H-pyrazole-4-carboxylic acid ethyl ester.

1H NMR(400MHz,DMSO-d6)：δ ppm 1.26(t,J=7.0Hz,3H)4.18(q,J=7.0Hz,2H),5.22(s,2H),6.47(br s,2H),6.64(d,J=8.3Hz,1H),7.41(d,J=2.2Hz,1H),7.55(s,1H),7.67(d,J=1.7Hz,1H)。 1 H NMR (400MHz, DMSO- d6 ): δ ppm 1.26 (t, J = 7.0Hz, 3H) 4.18 (q, J = 7.0Hz, 2H), 5.22 (s, 2H), 6.47 (br s, 2H) , 6.64 (d, J = 8.3 Hz, 1H), 7.41 (d, J = 2.2 Hz, 1H), 7.55 (s, 1H), 7.67 (d, J = 1.7 Hz, 1H).

MS：m/z 314.0(M+H)+。 MS: m/z 314.0 (M + H) + .

步驟3：製備5-氯-1'-(2,4-二氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 Step 3: Preparation of 5-chloro-1'-(2,4-dichlorobenzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b Pyridine]-2,6'(1'H)-dione

在室溫下，向5-胺基-1-(2,4-二氯苄基)-1H-吡唑-4-甲酸乙酯(3g，9.6mmol)於乙醇(15mL)中之溶液中添加水(15mL)，繼而添加氫氧化鈉(1.15g，28.75mmol)，且將反應混合物在80℃下維持4h。將 反應混合物冷卻至室溫且濃縮至乾燥以產生5-胺基-1-(2,4-二氯苄基)-1H-吡唑-4-甲酸鈉。在室溫下，向乙酸(15mL)中之所得鹽中添加5-氯吲哚啉-2,3-二酮(1.74g，9.585mmol)及麥氏酸(1.4g，9.585mmol)且在100℃下維持4h。將反應混合物冷卻至室溫且在減壓下移除溶劑。向反應混合物中添加冰水且在室溫下攪拌15min。將經過濾獲得之固體以水(2×15mL)洗滌且在真空下乾燥。藉由以丙酮(2×20mL)洗滌純化所得固體以提供3g(71.4%)呈白色固體狀之5-氯-1'-(2,4-二氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮。 Add to a solution of ethyl 5-amino-1-(2,4-dichlorobenzyl)-1H-pyrazole-4-carboxylate (3 g, 9.6 mmol) in ethanol (15 mL) Water (15 mL) followed by sodium hydroxide (1.15 g, 28.75 mmol) and the reaction mixture was maintained at &lt The reaction mixture was cooled to room temperature and concentrated to dryness to give 5-amino-l-(2,4-dichlorobenzyl)-1H-pyrazole-4-carboxylate . 5-Chloroporphyrin-2,3-dione (1.74 g, 9.585 mmol) and McDonald's (1.4 g, 9.585 mmol) were added to the resulting salt in acetic acid (15 mL) at rt. Maintain at °C for 4h. The reaction mixture was cooled to room temperature and the solvent was removed under reduced pressure. Ice water was added to the reaction mixture and stirred at room temperature for 15 min. The solid obtained by filtration was washed with water (2×15 mL) and dried under vacuum. The obtained solid was purified by washing with acetone (2×20 mL) to give 3 g (71.4%) of 5-chloro-1'-(2,4-dichlorobenzyl)-5',7'- Hydrogen spiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione .

1H NMR(400MHz,DMSO-d6)：δ ppm 2.55(d,J=15.8Hz,1H),3.14(d,J=15.8Hz,1H),5.36(q,J=16.7Hz,2H),6.78(d,J=8.4Hz,1H),6.85(s,1H),6.92(d,J=8.4Hz,1H),7.32(dd,J=1.7,6.2Hz,1H),7.35(s,1H),7.46(dd,J=1.8,6.5Hz,1H),7.68(d,J=1.8Hz,1H),10.64(s,1H),11.10(s,1H)。 1 H NMR (400MHz, DMSO- d6 ): δ ppm 2.55 (d, J = 15.8Hz, 1H), 3.14 (d, J = 15.8Hz, 1H), 5.36 (q, J = 16.7Hz, 2H), 6.78 (d, J = 8.4 Hz, 1H), 6.85 (s, 1H), 6.92 (d, J = 8.4 Hz, 1H), 7.32 (dd, J = 1.7, 6.2 Hz, 1H), 7.35 (s, 1H) , 7.46 (dd, J = 1.8, 6.5 Hz, 1H), 7.68 (d, J = 1.8 Hz, 1H), 10.64 (s, 1H), 11.10 (s, 1H).

MS=447.0(M+H)+。 MS = 447.0 (M+H) + .

步驟4：製備(R)-5-氯-1'-(2,4-二氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 Step 4: Preparation of (R)-5-chloro-1'-(2,4-dichlorobenzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3 ,4-b]pyridine]-2,6'(1'H)-dione

使2g 5-氯-1'-(2,4-二氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮經受對掌性製備型HPLC(管柱：Chiralpak IA(4.6×250mm)5μ；移動相：己烷：EtOH：CHCl3：TFA(70：15：15：0.1)；流速：1.0mL/min；稀釋劑：移動相；UV：255nm)以提供呈白色固體狀之兩種對映異構體：208mg(R)-5-氯-1'-(2,4-二氯 苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮(實例2)(峰-1，8.43min於對掌性HPLC中，98.8% ee)與180mg(S)-5-氯-1'-(2,4-二氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮(峰-2，12.63min於對掌性HPLC中，99% ee)。實例2之特徵資料展示如下：[α]27.5 D：-123.1(c=0.56,DMSO)。 2g 5-chloro-1'-(2,4-dichlorobenzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine ]-2,6'(1'H)-dione was subjected to palmar preparative HPLC (column: Chiralpak IA (4.6 x 250 mm) 5μ; mobile phase: hexane: EtOH: CHCl 3 : TFA (70:15) : 15:0.1); flow rate: 1.0 mL/min; diluent: mobile phase; UV: 255 nm) to provide two enantiomers as a white solid: 208 mg (R)-5-chloro-1'- (2,4-Dichlorobenzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1' H)-dione (Example 2) (peak-1, 8.43 min in palmitic HPLC, 98.8% ee) and 180 mg (S)-5-chloro-1'-(2,4-dichlorobenzyl) -5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione (peak-2, 12.63 min in palmar HPLC, 99% ee). The characteristics of Example 2 are shown below: [α] 27.5 D : -123.1 (c = 0.56, DMSO).

1H NMR(400MHz,DMSO-d6)：δ ppm 2.55(d,J=15.8.0Hz,1H),3.14(d,J=15.8Hz,1H),5.36(q,J=16.7Hz,2H),6.78(d,J=8.4Hz,1H),6.85(s,1H),6.92(d,J=8.4Hz,1H),7.32(dd,J=1.7,6.2Hz,1H),7.35(s,1H),7.46(dd,J=1.8,6.5Hz,1H),7.68(d,J=1.8Hz,1H),10.64(s,1H),11.10(s,1H)。 1 H NMR (400MHz, DMSO- d6 ): δ ppm 2.55 (d, J = 15.8.0Hz, 1H), 3.14 (d, J = 15.8Hz, 1H), 5.36 (q, J = 16.7Hz, 2H), 6.78 (d, J = 8.4 Hz, 1H), 6.85 (s, 1H), 6.92 (d, J = 8.4 Hz, 1H), 7.32 (dd, J = 1.7, 6.2 Hz, 1H), 7.35 (s, 1H) ), 7.46 (dd, J = 1.8, 6.5 Hz, 1H), 7.68 (d, J = 1.8 Hz, 1H), 10.64 (s, 1H), 11.10 (s, 1H).

MS=447.0(M+H)+。 MS = 447.0 (M+H) + .

實例3-1Example 3-1 製備(R)-5-氯-1'-(4-氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 Preparation of (R)-5-chloro-1'-(4-chlorobenzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine ]-2,6'(1'H)-dione

步驟1：製備(4-氯苄基)肼 Step 1: Preparation of (4-chlorobenzyl) hydrazine

將1-氯-4-(氯甲基)苯(6g，37.0mmol)於乙醇(30mL)中之溶液逐滴添加至水合肼(18.5g，370mmol)於乙醇(40mL)中之溶液中且在室 溫下攪拌24h。反應完成後，蒸發過量乙醇及肼。向上述反應混合物中添加50%氫氧化鈉溶液(50mL)於水中之溶液。以***(3×40mL)萃取水溶液。使經合併之有機層經無水Na2SO4乾燥且在減壓下濃縮以提供呈無色液體狀之(4-氯苄基)肼，其不經任何進一步純化即用於下一步驟。 A solution of 1-chloro-4-(chloromethyl)benzene (6 g, 37.0 mmol) in ethanol (30 mL) was added dropwise to a solution of hydrazine hydrate (18.5 g, 370 mmol) in ethanol (40 mL) and Stir at room temperature for 24 h. After the reaction was completed, excess ethanol and hydrazine were evaporated. A solution of 50% sodium hydroxide solution (50 mL) in water was added to the above reaction mixture. The aqueous solution was extracted with diethyl ether (3×40 mL). The combined so that the organic layer was dried over anhydrous Na 2 SO 4 and concentrated to provide a colorless liquid of (4-chlorobenzyl) hydrazine under reduced pressure, which was used without any further purification in the next step.

步驟2：製備5-胺基-1-(4-氯苄基)-1H-吡唑-4-甲酸乙酯 Step 2: Preparation of ethyl 5-amino-1-(4-chlorobenzyl)-1H-pyrazole-4-carboxylate

將(E)-2-氰基-3-乙氧基丙烯酸乙酯(1.64g，9.708mmol)及(4-氯苄基)肼(2g，9.708mmol)於乙醇中之溶液在80℃下攪拌隔夜。在減壓下移除溶劑。藉由使用石油醚中20%乙酸乙酯之溶劑梯度作為溶離劑，經矽膠(100-200目)管柱層析純化粗化合物以提供400mg(11%)呈奶白色固體狀之5-胺基-1-(4-氯苄基)-1H-吡唑-4-甲酸乙酯。 A solution of (E)-2-cyano-3-ethoxy acrylate (1.64 g, 9.708 mmol) and (4-chlorobenzyl) hydrazine (2 g, 9.708 mmol) in ethanol was stirred at 80 ° C. Overnight. The solvent was removed under reduced pressure. By using a solvent gradient of 20% ethyl acetate in petroleum ether as the eluent, dried over silica gel (100-200 mesh) The crude compound was purified by column chromatography to provide 400mg (11%) as a white solid of 5-amino-milk Ethyl 1-(4-chlorobenzyl)-1H-pyrazole-4-carboxylate .

1H NMR(400MHz,DMSO-d6)：1.24(t,J=7.0Hz,3H),4.16(q,J=7.0Hz,2H),5.17(s,2H),6.41(br s,2H),7.16(d,J=8.4Hz,2H),7.40(d,J=8.4Hz,2H),7.50(s,1H)。 1 H NMR (400MHz, DMSO- d 6): 1.24 (t, J = 7.0Hz, 3H), 4.16 (q, J = 7.0Hz, 2H), 5.17 (s, 2H), 6.41 (br s, 2H) , 7.16 (d, J = 8.4 Hz, 2H), 7.40 (d, J = 8.4 Hz, 2H), 7.50 (s, 1H).

LC-MS：m/z 280.1(M+H)+。 LC-MS: m / z 280.1 (M + H) +.

步驟3：製備5-氯-1'-(4-氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 Step 3: Preparation of 5-chloro-1 '-(4-chlorobenzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine] -2,6'(1'H)-dione

在室溫下，向5-胺基-1-(4-氯苄基)-1H-吡唑-4-甲酸乙酯(400 mg，1.433mmol)於乙醇(5mL)中之溶液中添加水(5mL)，繼而添加氫氧化鈉(150mg，2.867mmol)，且將所得反應混合物在90℃下維持6h。將所得反應混合物冷卻至室溫且濃縮至乾燥以產生5-胺基-1-(4-氯苄基)-1H-吡唑-4-甲酸鈉。在室溫下，向乙酸中之所得鹽中添加5-氯吲哚啉-2,3-二酮(240mg，1.576mmol)及麥氏酸(237mg，1.648mmol)且在100℃下攪拌3h。將反應混合物冷卻至室溫且在減壓下移除溶劑。向反應混合物中添加水且在室溫下攪拌15min。將經過濾獲得之固體以水(2×15mL)洗滌且在真空下乾燥。以丙酮洗滌所得固體以產生純的5-氯-1'-(4-氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮550mg(92%)。 To 5-Amino-1-(4-chlorobenzyl)-1H-pyrazole-4-carboxylic acid ethyl ester (400 at room temperature) Water (5 mL) was added to a solution of EtOAc (EtOAc) (EtOAc). The resulting reaction mixture was cooled to room temperature and concentrated to dryness to give 5-amino-l-(4-chlorobenzyl)-1H-pyrazole-4-carboxylate. 5-Chloroporphyrin-2,3-dione (240 mg, 1.576 mmol) and McDonald's (237 mg, 1.648 mmol) were added to the obtained salt in acetic acid at room temperature and stirred at 100 ° C for 3 h. The reaction mixture was cooled to room temperature and the solvent was removed under reduced pressure. Water was added to the reaction mixture and stirred at room temperature for 15 min. The solid obtained by filtration was washed with water (2×15 mL) and dried under vacuum. The resulting solid was washed with acetone to give pure 5-chloro-1 '-(4-chlorobenzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazole[3,4 -b]pyridine]-2,6'(1'H)-dione 550 mg (92%).

1H NMR(400MHz,DMSO-d6)：δ ppm 2.51(d,J=13.6Hz,1H),3.12(d,J=15.8Hz,1H),5.28(q,J=15.4Hz,10.6Hz,2H),6.79(s,1H),6.9(d,J=7.9Hz,1H),7.35-7.2(m,4H),7.42(d,J=8.3Hz,2H),10.59(br s,1H),11.08(br s,1H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm 2.51 (d, J = 13.6Hz, 1H), 3.12 (d, J = 15.8Hz, 1H), 5.28 (q, J = 15.4Hz, 10.6Hz, 2H), 6.79 (s, 1H), 6.9 (d, J = 7.9 Hz, 1H), 7.35-7.2 (m, 4H), 7.42 (d, J = 8.3 Hz, 2H), 10.59 (br s, 1H) , 11.08 (br s, 1H).

LC-MS：m/z 413.0(M+H)+。 LC-MS: m / z 413.0 (M + H) +.

步驟4：製備(R)-5-氯-1'-(4-氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 Step 4: Preparation of (R)-5-chloro-1'-(4-chlorobenzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4- b]pyridine]-2,6'(1'H)-dione

使500mg 5-氯-1'-(4-氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮經受對掌性製備型HPLC(管柱：Chiralpak IA(4.6×250mm)5μ；移動相：己烷：EtOH：DEA(60：40：0.1)；流速：1.0mL/min；UV：255nm)以提供呈白色固體狀之兩種對映異構體： 110mg(R)-5-氯-1'-(4-氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮(實例3-1)(峰-1，6.87min於對掌性HPLC中，99.9% ee)與150mg(S)-5-氯-1'-(4-氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮(峰-2，8.16min於對掌性HPLC中，99% ee)。實例3-1之特徵資料展示如下：[α]25.4 D：-121.7(c=0.25,MeOH)。 500 mg 5-chloro-1 '-(4-chlorobenzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2 , 6'(1'H)-dione was subjected to palmar preparative HPLC (column: Chiralpak IA (4.6 x 250 mm) 5 μ; mobile phase: hexane: EtOH: DEA (60: 40: 0.1); flow rate: 1.0 mL/min; UV: 255 nm) to provide the two enantiomers as a white solid: 110 mg (R)-5-chloro-1 '-(4-chlorobenzyl)-5', 7'- Dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione (Example 3-1) (peak-1, 6.87) Min in palmitic HPLC, 99.9% ee) and 150 mg (S)-5-chloro-1'-(4-chlorobenzyl)-5',7'-dihydrospiro[porphyrin-3,4 '-Pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione (peak-2, 8.16 min in palmitic HPLC, 99% ee). The characteristics of Example 3-1 are shown below: [α] 25.4 D : -121.7 (c = 0.25, MeOH).

1H NMR(400MHz,DMSO-d6)：δ ppm 2.51(d,J=13.6Hz,1H),3.12(d,J=15.8Hz,1H),5.28(q,J=15.4Hz,10.6Hz,2H),6.79(s,1H),6.9(d,J=7.9Hz,1H),7.35-7.2(m,4H),7.42(d,J=8.3Hz,2H),10.59(br s,1H),11.08(br s,1H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm 2.51 (d, J = 13.6Hz, 1H), 3.12 (d, J = 15.8Hz, 1H), 5.28 (q, J = 15.4Hz, 10.6Hz, 2H), 6.79 (s, 1H), 6.9 (d, J = 7.9 Hz, 1H), 7.35-7.2 (m, 4H), 7.42 (d, J = 8.3 Hz, 2H), 10.59 (br s, 1H) , 11.08 (br s, 1H).

LC-MS：m/z 413.0(M+H)+。 LC-MS: m / z 413.0 (M + H) +.

使用通用程序以及上述實例之程序，由適當起始物質來製備以下化合物。 The following compounds were prepared from the appropriate starting materials using the general procedures and procedures of the above examples.

實例3-2Example 3-2 (R)-5-氯-1'-(4-(三氟甲基)苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-chloro-1'-(4-(trifluoromethyl)benzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4 -b]pyridine]-2,6'(1'H)-dione

1H-NMR(DMSO-d6)：δ ppm 2.66(d,1H，與DMSO合併)。3.12-3.16(d,J=15.8Hz,1H),5.40(q,J=16.2Hz,2H),6.82(s,1H),6.91(d,J=8.3Hz,1H),7.29-7.33(m,2H),7.42(d,J=7.92Hz,1H),7.74(d,J=7.9Hz,2H),10.62(s,1H),11.14(br s,1H)。 1 H-NMR (DMSO-d 6 ): δ ppm 2.66 (d, 1H, combined with DMSO). 3.12-3.16(d, J = 15.8 Hz, 1H), 5.40 (q, J = 16.2 Hz, 2H), 6.82 (s, 1H), 6.91 (d, J = 8.3 Hz, 1H), 7.29-7.33 (m , 2H), 7.42 (d, J = 7.92 Hz, 1H), 7.74 (d, J = 7.9 Hz, 2H), 10.62 (s, 1H), 11.14 (br s, 1H).

MS：m/z 445.0(M-H)-。 MS: m/z 445.0 (MH) - .

實例3-3-1Example 3-3-1 (R)-5-氯-1'-(1-(4-氯苯基)乙基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，異構體1。 (R)-5-chloro-1'-(1-(4-chlorophenyl)ethyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3, 4-b]pyridine]-2,6'(1'H)-dione, isomer 1.

1H NMR(400MHz,DMSO-d6)：δ ppm 1.72(d,J=7.0Hz,3H),2.52(d,J=16.3Hz,1H),3.07(d,J=16.6Hz,1H),5.66(d,J=7.0Hz,1H),6.83(s,1H),6.90(d,J=8.8Hz,1H),7.28-7.42(m,6H),10.59(s,1H),11.05(br s,1H)。 1 H NMR (400MHz, DMSO- d6 ): δ ppm 1.72 (d, J = 7.0Hz, 3H), 2.52 (d, J = 16.3Hz, 1H), 3.07 (d, J = 16.6Hz, 1H), 5.66 (d, J = 7.0 Hz, 1H), 6.83 (s, 1H), 6.90 (d, J = 8.8 Hz, 1H), 7.28-7.42 (m, 6H), 10.59 (s, 1H), 11.05 (br s , 1H).

MS：m/z 427.0(M+H)+。 MS: m/z 427.0 (M + H) + .

實例3-3-2Example 3-3-2 (S)-5-氯-1'-(1-(4-氯苯基)乙基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，異構體1。 (S)-5-chloro-1'-(1-(4-chlorophenyl)ethyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3, 4-b]pyridine]-2,6'(1'H)-dione, isomer 1.

1H NMR(400MHz,DMSO-d6)：δ ppm 1.72(d,J=7.0Hz,3H),2.52(d,J=16.3Hz,1H),3.07(d,J=16.6Hz,1H),5.66(d,J=7.0Hz,1H),6.83(s,1H),6.90(d,J=8.8Hz,1H),7.28-7.42(m,6H),10.59(s,1H),11.05(br s,1H)。 1 H NMR (400MHz, DMSO- d6 ): δ ppm 1.72 (d, J = 7.0Hz, 3H), 2.52 (d, J = 16.3Hz, 1H), 3.07 (d, J = 16.6Hz, 1H), 5.66 (d, J = 7.0 Hz, 1H), 6.83 (s, 1H), 6.90 (d, J = 8.8 Hz, 1H), 7.28-7.42 (m, 6H), 10.59 (s, 1H), 11.05 (br s , 1H).

MS：m/z 427.0(M+H)+。 MS: m/z 427.0 (M + H) + .

藉由對掌性HPLC(管柱：Acquity；UPLC；BEH；C18；100×2.1mm、1.7μ；移動相：於水中之0.025% TFA及於CAN中之0.025% TFA；流速：0.4ml/min；稀釋劑：ACN；UV：214nm、254nm)，由5-氯-1'-(1-(4-氯苯基)乙基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮之外消旋混合物分別來解析實例3-3-1與3-3-2。 By palmar HPLC (column: Acquity; UPLC; BEH; C18; 100 x 2.1 mm, 1.7 μ; mobile phase: 0.025% TFA in water and 0.025% TFA in CAN; flow rate: 0.4 ml/min Diluent: ACN; UV: 214 nm, 254 nm) from 5-chloro-1'-(1-(4-chlorophenyl)ethyl)-5',7'-dihydrospiro[porphyrin-3 , 4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione racemic mixture was used to resolve Examples 3-3-1 and 3-3-2, respectively.

實例3-4Example 3-4 (R)-1'-(4-氯苄基)-5-(三氟甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-1'-(4-chlorobenzyl)-5-(trifluoromethyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4 -b]pyridine]-2,6'(1'H)-dione

1H-NMR(DMSO-d6)：δ ppm 2.57(d,1H，與DMSO合併)，3.23(d,J=16.0Hz,1H),5.27(q,J=16.0Hz,2H),6.77(s,1H),7.07(d,J=7.6Hz,1H),7.26(d,J=8.4Hz,2H),7.42(d,J=8.4Hz,2H),7.62(d,J=9.6Hz,2H),10.8(s,1H),11.1(br s,1H)。 1 H-NMR (DMSO-d 6 ): δ ppm 2.57 (d, 1H, combined with DMSO), 3.23 (d, J = 16.0 Hz, 1H), 5.27 (q, J = 16.0 Hz, 2H), 6.77 ( s, 1H), 7.07 (d, J = 7.6 Hz, 1H), 7.26 (d, J = 8.4 Hz, 2H), 7.42 (d, J = 8.4 Hz, 2H), 7.62 (d, J = 9.6 Hz, 2H), 10.8 (s, 1H), 11.1 (br s, 1H).

MS：m/z 447.0(M+H)+。 MS: m/z 447.0 (M + H) + .

實例3-5Example 3-5 (R)-5,6-二氯-1'-(4-氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5,6-dichloro-1'-(4-chlorobenzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b Pyridine]-2,6'(1'H)-dione

1H NMR(400MHz,DMSO-d6)：δ ppm 11.09(s,1H),10.72(s,1H),7.55(s,1H),7.43(d,J=8.8Hz,2H),7.26(d,J=8.4Hz,2H),7.08(s,1H),6.85(s,1H),5.27(q,J=14.6Hz,2H),3.18(d,J=16.0Hz,1H),2.54(d,J=16.0Hz,1H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm 11.09 (s, 1H), 10.72 (s, 1H), 7.55 (s, 1H), 7.43 (d, J = 8.8Hz, 2H), 7.26 (d , J = 8.4 Hz, 2H), 7.08 (s, 1H), 6.85 (s, 1H), 5.27 (q, J = 14.6 Hz, 2H), 3.18 (d, J = 16.0 Hz, 1H), 2.54 (d , J = 16.0 Hz, 1H).

MS：m/z 449.0(M+H)+。 MS: m/z 449.0 (M + H) + .

實例3-6Example 3-6 (R)-5-溴-1'-(4-氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-bromo-1'-(4-chlorobenzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine] -2,6'(1'H)-dione

1H NMR(400MHz,DMSO-d6)：δ ppm 2.52(d,J=14.1Hz,1H),3.12(d,J=15.8Hz,1H),5.27(q,J=15.8Hz,2H),6.79(s,1H),6.86(d,J=8.8Hz,1H),7.25(d,J=8.4Hz,2H),7.43-7.41(m,4H),10.59(s,1H),11.00(br s,1H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm 2.52 (d, J = 14.1Hz, 1H), 3.12 (d, J = 15.8Hz, 1H), 5.27 (q, J = 15.8Hz, 2H), 6.79 (s, 1H), 6.86 (d, J = 8.8 Hz, 1H), 7.25 (d, J = 8.4 Hz, 2H), 7.43 - 7.41 (m, 4H), 10.59 (s, 1H), 11.00 (br) s, 1H).

MS：m/z 457.0(M+H)+。 MS: m/z 457.0 (M + H) + .

實例3-7Example 3-7 (R)-1'-(4-氯苄基)-5-(三氟甲氧基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-1'-(4-chlorobenzyl)-5-(trifluoromethoxy)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3, 4-b]pyridine]-2,6'(1'H)-dione

1H-NMR(DMSO-d6)：δ ppm 2.56(d,1H，與DMSO合併)，3.16(d,J=15.8Hz,1H),5.27(q,J=15.9Hz,2H),6.76(s,1H),6.97(d,J=8.4Hz,1H),7.26(d,J=8.3Hz,2H),7.31(s,2H),7.43(d,J=8.4Hz,1H),10.66(s,1H),11.11(br s,1H)。 1 H-NMR (DMSO-d 6 ): δ ppm 2.56 (d, 1H, combined with DMSO), 3.16 (d, J = 15.8 Hz, 1H), 5.27 (q, J = 15.9 Hz, 2H), 6.76 ( s, 1H), 6.97 (d, J = 8.4 Hz, 1H), 7.26 (d, J = 8.3 Hz, 2H), 7.31 (s, 2H), 7.43 (d, J = 8.4 Hz, 1H), 10.66 ( s, 1H), 11.11 (br s, 1H).

MS m/z：463.3(M+H)+。 MS m/z: 463.3 (M + H) + .

實例3-8Example 3-8 (R)-6-氯-1'-(4-氯苄基)-5-氟-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-6-chloro-1'-(4-chlorobenzyl)-5-fluoro-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4- b]pyridine]-2,6'(1'H)-dione

1H NMR(400MHz,DMSO-d6)：δ ppm 11.10(s,1H),10.60(s,1H),7.42(m,3H),7.26(d,J=8.4Hz,2H),7.01(d,J=6Hz,1H),6.83(s,1H),5.27(q,J=14.2Hz,2H),3.15(d,J=16.0Hz,1H),2.55(d,1H，與DMSO合併)。 1 H NMR (400MHz, DMSO- d 6): δ ppm 11.10 (s, 1H), 10.60 (s, 1H), 7.42 (m, 3H), 7.26 (d, J = 8.4Hz, 2H), 7.01 (d , J = 6 Hz, 1H), 6.83 (s, 1H), 5.27 (q, J = 14.2 Hz, 2H), 3.15 (d, J = 16.0 Hz, 1H), 2.55 (d, 1H, combined with DMSO).

MS：m/z 432.0(M+H)+。 MS: m/z 432.0 (M + H) + .

實例3-9Example 3-9 (R)-5-氯-1'-(4-氯苄基)-7'-異丙基-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-chloro-1'-(4-chlorobenzyl)-7'-isopropyl-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3 ,4-b]pyridine]-2,6'(1'H)-dione

1H NMR(400MHz,DMSO-d6)：δ ppm1.39(m,6H),2.5(d，與DMSO合併，1H),3.21(d,J=15.6Hz,1H),3.92(m,1H),5.43(q,J=17.2Hz,2H),6.91-6.93(m,2H),7.13(d,J=8.4Hz,2H),7.34-7.31(m,2H),7.46(d,J=8.4Hz,2H),10.66(s,1H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm1.39 (m, 6H), 2.5 (d, merged with DMSO, 1H), 3.21 (d, J = 15.6Hz, 1H), 3.92 (m, 1H ), 5.43 (q, J = 17.2 Hz, 2H), 6.91-6.93 (m, 2H), 7.13 (d, J = 8.4 Hz, 2H), 7.34 - 7.31 (m, 2H), 7.46 (d, J = 8.4 Hz, 2H), 10.66 (s, 1H).

MS：m/z 455.1(M+H)+。 MS: m/z 455.1 (M + H) + .

實例3-10Example 3-10 (R)-5-氯-1'-(4-氯苄基)-7'-甲基-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-chloro-1'-(4-chlorobenzyl)-7'-methyl-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3, 4-b]pyridine]-2,6'(1'H)-dione

1H NMR(400MHz,DMSO-d6)：δ ppm2.63(d,J=15.8Hz,1H),3.21(d,J=15.4Hz,1H),3.26(s,3H),5.54(s,2H),6.95-6.90(m,2H),7.17(d,J=7.5Hz,2H),7.35-7.31(m,2H),7.44(d,J=7.9Hz,2H),10.68(s,1H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm2.63 (d, J = 15.8Hz, 1H), 3.21 (d, J = 15.4Hz, 1H), 3.26 (s, 3H), 5.54 (s, 2H), 6.95-6.90 (m, 2H), 7.17 (d, J = 7.5 Hz, 2H), 7.35-7.31 (m, 2H), 7.44 (d, J = 7.9 Hz, 2H), 10.68 (s, 1H) ).

MS：m/z 427.1(M+H)+。 MS: m/z 427.1 (M + H) + .

實例3-11Example 3-11 (R)-5,6-二氯-1'-(4-(三氟甲基)苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5,6-dichloro-1'-(4-(trifluoromethyl)benzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[ 3,4-b]pyridine]-2,6'(1'H)-dione

[α]25.9 D：-104.9(c=0.54,CHCl3)。 [α] 25.9 D : -104.9 (c = 0.54, CHCl 3 ).

1H-NMR(400MHz,DMSO-d6)：δ ppm 2.44(d,1H，與DMSO合併)，3.19(d,J=15.6Hz,1H),5.38(q,J=16.0Hz,2H),6.88(s,1H),7.09(s,1H),7.42(d,J=8.0Hz,2H),7.57(s,1H),7.74(d,J=8.4Hz,2H),10.74(s,1H),11.13(br s,1H)。 1 H-NMR (400MHz, DMSO -d 6): δ ppm 2.44 (d, 1H, merged with DMSO), 3.19 (d, J = 15.6Hz, 1H), 5.38 (q, J = 16.0Hz, 2H), 6.88 (s, 1H), 7.09 (s, 1H), 7.42 (d, J = 8.0 Hz, 2H), 7.57 (s, 1H), 7.74 (d, J = 8.4 Hz, 2H), 10.74 (s, 1H) ), 11.13 (br s, 1H).

MS：m/z 479.0(M-H)-。 MS: m/z 479.0 (MH) - .

實例3-12Example 3-12 (R)-5-(三氟甲基)-1'-(4-(三氟甲基)苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-(trifluoromethyl)-1'-(4-(trifluoromethyl)benzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazole And [3,4-b]pyridine]-2,6'(1'H)-dione

[α]25.0 D：-109.2(c=0.5,DMSO)。 [α] 25.0 D : -109.2 (c = 0.5, DMSO).

1H-NMR(400MHz,DMSO-d6)：δ ppm 2.46(d,1H，與DMSO合併)，3.24(d,J=15.6Hz及1H),5.41(q,J=16.4Hz,2H),6.81(s,1H),7.08(d,J=8.0Hz,1H),7.43(d,J=8.0Hz),7.627-7.647(m,2H),10.88(s,1H),11.14(br s,1H)。 1 H-NMR (400MHz, DMSO -d 6): δ ppm 2.46 (d, 1H, merged with DMSO), 3.24 (d, J = 15.6Hz and 1H), 5.41 (q, J = 16.4Hz, 2H), 6.81 (s, 1H), 7.08 (d, J = 8.0 Hz, 1H), 7.43 (d, J = 8.0 Hz), 7.627-7.647 (m, 2H), 10.88 (s, 1H), 11.14 (br s, 1H).

MS：m/z 481.1(M+H)+。 MS: m/z 481.1 (M + H) + .

實例3-13Example 3-13 (R)-5-氯-6-氟-1'-(4-(三氟甲基)苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-chloro-6-fluoro-1'-(4-(trifluoromethyl)benzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazole [3,4-b]pyridine]-2,6'(1'H)-dione

[α]28.3 D：-103.2(c=0.53,MeOH：CHCl3=1：1)。 [α] 28.3 D : -103.2 (c = 0.53, MeOH: CHCl 3 = 1:1).

1H-NMR(400MHz,DMSO-d6)：δ ppm 2.59(d,1H，與DMSO合併)。3.18(d,J=16.4Hz,1H),5.38(q,J=16.4Hz,2H),6.85(s,1H),6.93(d,J=9.6Hz,1H),748(m,3H),7.74(d,J=8.4Hz,2H),10.74(s,1H),11.11(br s,1H)。 1 H-NMR (400 MHz, DMSO-d 6 ): δ </ RTI></RTI> 2.59 (d, 1H, combined with DMSO). 3.18(d, J = 16.4 Hz, 1H), 5.38 (q, J = 16.4 Hz, 2H), 6.85 (s, 1H), 6.93 (d, J = 9.6 Hz, 1H), 748 (m, 3H), 7.74 (d, J = 8.4 Hz, 2H), 10.74 (s, 1H), 11.11 (br s, 1H).

MS：m/z 465.1(M+H)+。 MS: m/z 465.1 (M + H) + .

實例3-14Example 3-14 (R)-5,6-二氯-1'-((5-氯吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5,6-Dichloro-1'-((5-chloropyridin-2-yl)methyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazole And [3,4-b]pyridine]-2,6'(1'H)-dione

1H NMR(400MHz,DMSO-d6)：δ ppm,2.56(d,1H，與DMSO合併)，3.19(d,J=15.9Hz,1H),5.40(q,J=16.3Hz,2H),6.87(s,1H),7.06(d,J=8.3Hz,1H),7.10(s,1H),7.57(s,1H),7.94(dd,J=2.6、8.3 Hz,1H),8.6(d,J=1.8Hz,1H),10.7(s,1H),11.07(br s,1H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm, 2.56 (d, 1H, merged with DMSO), 3.19 (d, J = 15.9Hz, 1H), 5.40 (q, J = 16.3Hz, 2H), 6.87 (s, 1H), 7.06 (d, J = 8.3 Hz, 1H), 7.10 (s, 1H), 7.57 (s, 1H), 7.94 (dd, J = 2.6, 8.3 Hz, 1H), 8.6 (d , J = 1.8 Hz, 1H), 10.7 (s, 1H), 11.07 (br s, 1H).

MS：m/z 448.0(M+H)+。 MS: m / z 448.0 (M + H) +.

實例3-15Example 3-15 (R)-5-氯-1'-((5-氯吡啶-2-基)甲基)-6-氟-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-chloro-1'-((5-chloropyridin-2-yl)methyl)-6-fluoro-5',7'-dihydrospiro[porphyrin-3,4'-pyridyl Oxazo[3,4-b]pyridine]-2,6'(1'H)-dione

1H NMR(400MHz,DMSO-d 6)d ppm 11.06(s,1H),10.77(s,1H),8.62(d,J=2.4Hz,1H),7.94(dd,J=2.4,8.4Hz,1H),7.51(d,J=7.2Hz,1H),7.06(d,J=8.8Hz,1H),6.94(d,J=9.2Hz,1 H),6.85(s,1H),5.40(q,J=14.8Hz,2H),3.18(d,J=16Hz,1H),2.55(d，與DMSO溶劑峰重疊)。 1 H NMR (400 MHz, DMSO- d 6 ) d ppm 11.06 (s, 1H), 10.77 (s, 1H), 8.62 (d, J = 2.4 Hz, 1H), 7.94 (dd, J = 2.4, 8.4 Hz, 1H), 7.51 (d, J = 7.2 Hz, 1H), 7.06 (d, J = 8.8 Hz, 1H), 6.94 (d, J = 9.2 Hz, 1 H), 6.85 (s, 1H), 5.40 (q) , J = 14.8 Hz, 2H), 3.18 (d, J = 16 Hz, 1H), 2.55 (d, overlap with the DMSO solvent peak).

實例3-16Example 3-16 (R)-1'-((5-氯吡啶-2-基)甲基)-5-(三氟甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-1'-((5-chloropyridin-2-yl)methyl)-5-(trifluoromethyl)-5',7'-dihydrospiro[porphyrin-3,4'- Pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione

1H NMR(400MHz,DMSO-d6)：δ ppm 2.58(d,1H，與DMSO合併)，3.22(d,J=15.8Hz,1H),5.42(q,J=16.4Hz,2H),6.80(s,1H),7.08 (d,J=7.9Hz,2H),7.63(d,J=8.8Hz,2H),7.94(d,J=2.8Hz,1H),8.61(br,s,1H),10.89(br,s,1H),δ 11.07(br,s,1H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm 2.58 (d, 1H, merged with DMSO), 3.22 (d, J = 15.8Hz, 1H), 5.42 (q, J = 16.4Hz, 2H), 6.80 (s, 1H), 7.08 (d, J = 7.9 Hz, 2H), 7.63 (d, J = 8.8 Hz, 2H), 7.94 (d, J = 2.8 Hz, 1H), 8.61 (br, s, 1H) , 10.89 (br, s, 1H), δ 11.07 (br, s, 1H).

MS：m/z 448.1(M+H)+。 MS: m/z 448.1 (M + H) + .

實例3-17Example 3-17 (R)-5-氯-1'-((5-氯吡啶-2-基)甲基)-1-環丙基-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-chloro-1'-((5-chloropyridin-2-yl)methyl)-1-cyclopropyl-5',7'-dihydrospiro[porphyrin-3,4' -pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione

1H NMR(400MHz,DMSO-d6)：δ ppm 0.7-0.84(m,2H),0.87-0.99(m,2H),2.57(d,1H，與DMSO合併)，2.70(m,1H),3.10(d,J=16.4Hz,1H),5.35(q,J=9.6Hz,2H),6.78(s,1H),7.05(d,J=8.4Hz,1H),7.18(d,J=8.4Hz,1H),7.35(d,J=1.8Hz,1H),7.41-7.45(m,2H),7.92-7.94(dd,J=2.0,1.2Hz,1H)8.60(s,1H),δ 11.05(br s,1H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm 0.7-0.84 (m, 2H), 0.87-0.99 (m, 2H), 2.57 (d, 1H, merged with DMSO), 2.70 (m, 1H) , 3.10(d, J = 16.4 Hz, 1H), 5.35 (q, J = 9.6 Hz, 2H), 6.78 (s, 1H), 7.05 (d, J = 8.4 Hz, 1H), 7.18 (d, J = 8.4) Hz, 1H), 7.35 (d, J = 1.8 Hz, 1H), 7.41-7.45 (m, 2H), 7.92-7.94 (dd, J = 2.0, 1.2 Hz, 1H) 8.60 (s, 1H), δ 11.05 (br s, 1H).

MS：m/z 454.1(M+H)+。 MS: m/z 454.1 (M + H) + .

實例4-1、4-2、4-3及4-4Examples 4-1, 4-2, 4-3, and 4-4 製備5'-氯-3-(1-(4-氯苯基)乙基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮之四種異構體。 Preparation of 5'-chloro-3-(1-(4-chlorophenyl)ethyl)-4,6-dihydrospiro[[1,2,3]triazolo[4,5-b]pyridine-7 , 4'-porphyrin]-2', 5 (3H)-dione of four isomers.

步驟1：製備1-(4-氯苯基)乙醇Step 1: Preparation of 1-(4-chlorophenyl)ethanol

在0℃下，向4'-氯苯乙酮(20g，129.37mmol)於甲醇(250mL)中之冷卻溶液中以小份添加NaBH4(5.87g，155.24mmol)，且將所得混合物在室溫下攪拌4h。將反應混合物以1N HCl(100mL)中止且以CH2Cl2(3×200mL)萃取。使經合併之有機層經鹽水洗滌、經無水Na2SO4乾燥且在減壓下濃縮以提供19g呈無色液體狀之1-(4-氯苯基)乙醇 1(94%產率)。 At 0 ℃, to 4'-chloroacetophenone (20g, 129.37mmol) in methanol (250 mL) to the cooled solution in small portions was added NaBH 4 (5.87g, 155.24mmol), and the resulting mixture was at room temperature Stir under 4h. The reaction mixture was quenched and extracted with CH 2 Cl 2 (3 × 200mL ) in 1N HCl (100mL). The organic layers were washed with brine so that the combined sum, dried over anhydrous Na 2 SO 4 dried and concentrated to provide 19g of colorless liquid of 1- (4-Chlorophenyl) ethanol 1 (yield 94%) under reduced pressure.

1H-NMR(400MHz,CDCl3)；δ 7.31(s,4H),4.85-4.92(m,1H),1.79(br s,1H),1.47(d,J=6.8Hz,3H)。 1 H-NMR (400 MHz, CDCl 3 ); δ 7.31 (s, 4H), 4.85 - 4.92 (m, 1H), 1.79 (br s, 1H), 1.47 (d, J = 6.8 Hz, 3H).

步驟2：製備1-(1-溴乙基)-4-氯苯 Step 2: Preparation of 1-(1-bromoethyl)-4-chlorobenzene

在0℃下，向1-(4-氯苯基)乙醇(19g，121.33mmol)於CH2Cl2(200mL)中之冷卻溶液中緩慢添加三溴化磷(13.8mL，145.59mmol)且使所得混合物在室溫下攪拌4h。將反應團以CH2Cl2(500mL)稀釋，且以 水、飽和NaHCO3洗滌，繼而以水及鹽水洗滌。使有機層經無水Na2SO4乾燥且在減壓下濃縮以提供23.4g(87%)呈無色液體狀之1-(1-溴乙基)-4-氯苯。 At 0 ℃, a solution of 1- (4-chlorophenyl) ethanol (19g, 121.33mmol) in the in CH 2 Cl 2 (200mL) was cooled was slowly added phosphorous tribromide (13.8mL, 145.59mmol) and that the The resulting mixture was stirred at room temperature for 4 h. The reaction was diluted with radical CH 2 Cl 2 (500mL), and water, washed with saturated NaHCO 3, then with water and brine. The organic layer was dried over anhydrous Na 2 SO 4 and concentrated to provide 23.4g (87%) as a colorless liquid of 1- (1-bromoethyl) -4-chlorobenzene under reduced pressure.

1H-NMR(400MHz,DMSO-d 6 )；δ 7.52-7.55(m,2H),7.45-7.39(m,2H),5.51(q,J=6.9Hz,1H),1.97(d,J=6.8Hz,3H)。 1 H-NMR (400MHz, DMSO- d 6); δ 7.52-7.55 (m, 2H), 7.45-7.39 (m, 2H), 5.51 (q, J = 6.9Hz, 1H), 1.97 (d, J = 6.8 Hz, 3H).

步驟3：製備1-(1-疊氮基乙基)-4-氯苯 Step 3: Preparation of 1-(1-azidoethyl)-4-chlorobenzene

向1-(1-溴乙基)-4-氯苯(23.4g，106.6mmol)於DMSO(250mL)中之溶液中添加疊氮化鈉(6.93g，266.5mmol)且將所得混合物在室溫下攪拌5h。以水中止反應混合物且以乙酸乙酯(3×300mL)萃取。以水、鹽水洗滌經合併之有機層、經無水Na2SO4乾燥且在減壓下濃縮以提供16.5g(85%)呈無色液體狀之1-(1-疊氮基乙基)-4-氯苯。 Add sodium azide (6.93 g, 266.5 mmol) to a solution of 1-(1-bromoethyl)-4-chlorobenzene (23.4 g, 106.6 mmol) in EtOAc (250 mL). Stir under 5h. The reaction mixture was quenched with water and extracted with EtOAc EtOAc. Washed with water, brine and the organic layers were combined, dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure to provide 16.5g (85%) as a colorless liquid of 1- (1-azido-ethyl) -4 - Chlorobenzene .

1H-NMR(400MHz,DMSO-d 6 )；δ 7.38-7.48(m,4H),4.87(q,J=6.5Hz,1H),1.44(d,J=6.8Hz,3H)。 1 H-NMR (400 MHz, DMSO- d 6 ); δ 7.38-7.48 (m, 4H), 4.87 (q, J = 6.5 Hz, 1H), 1.44 (d, J = 6.8 Hz, 3H).

步驟4：製備5-胺基-1-(1-(4-氯苯基)乙基)-1H-1,2,3-***-4-甲酸乙酯 Step 4: Preparation of ethyl 5-amino-1-(1-(4-chlorophenyl)ethyl)-1H-1,2,3-triazole-4-carboxylate

向1-(1-疊氮基乙基)-4-氯苯(1g，5,51mmol)及氰基乙酸乙酯(0.88mL，8.26mmol)於乙醇(10mL)中之冷卻溶液中添加乙氧化鈉(21%於乙醇中)(4.1mL，12.66mmol)且將所得混合物在回流溫度下攪拌5h。在減壓下濃縮反應混合物且將殘餘物分配在乙酸乙酯與水之間。使經 合併之有機層經無水Na2SO4乾燥且濃縮。藉由使用石油醚中40-45% EtOAc之溶劑梯度作為溶離劑，經矽膠(100-200目)管柱層析純化粗產物以提供350mg(21%)呈黃色固體狀之5-胺基-1-(1-(4-氯苯基)乙基)-1H-1,2,3-***-4-甲酸乙酯。 Add ethoxylation to a cooled solution of 1-(1-azidoethyl)-4-chlorobenzene (1 g, 5,51 mmol) and ethyl cyanoacetate (0.88 mL, 8.26 mmol) in ethanol (10 mL) Sodium (21% in ethanol) (4.1 mL, 12.66 mmol) and mixture was stirred at reflux temperature for 5 h. The reaction mixture was concentrated under reduced pressure and the residue was partitioned betweenEtOAc and water. The combined so that the organic layer was dried over anhydrous Na 2 SO 4 and concentrated. By using a solvent gradient of 40-45% EtOAc in petroleum ether as the eluent, dried over silica gel (100-200 mesh) The crude product was purified by column chromatography to provide 350mg (21%) as a yellow solid of 5-amino - Ethyl 1-(1-(4-chlorophenyl)ethyl)-1H-1,2,3-triazole-4-carboxylate .

1H-NMR(400MHz,DMSO-d 6 )；δ 7.42(d,J=8.8Hz,2H),7.26(d,J=8.8Hz,2H),6.5(s,2H),5.72(q,J=7.0Hz,1H),4.25(q,J=7.2Hz,2H),1.84(d,J=7.2Hz,3H),1.27(t,J=7.0Hz,3H)；MS：m/z 295.0[M+H]+。 1 H-NMR (400 MHz, DMSO- d 6 ); δ 7.42 (d, J = 8.8 Hz, 2H), 7.26 (d, J = 8.8 Hz, 2H), 6.5 (s, 2H), 5.72 (q, J) =7.0 Hz, 1H), 4.25 (q, J = 7.2 Hz, 2H), 1.84 (d, J = 7.2 Hz, 3H), 1.27 (t, J = 7.0 Hz, 3H); MS: m/z 295.0 [ M+H] + .

步驟5：製備5-胺基-1-(1-(4-氯苯基)乙基)-1H-1,2,3-***-4-甲酸 Step 5: Preparation of 5-amino-1-(1-(4-chlorophenyl)ethyl)-1H-1,2,3-triazole-4-carboxylic acid

向5-胺基-1-(1-(4-氯苯基)乙基)-1H-1,2,3-***-4-甲酸乙酯(320mg，1.08mmol)於乙醇(3mL)中之溶液中添加NaOH(130mg，3.26mmol)於水(2mL)中之溶液且將所得混合物在60℃下攪拌4h。在減壓下濃縮反應混合物且將殘餘物溶於冷水(10mL)中且以飽和硫酸氫鈉酸化至pH約為6。藉由過濾收集所得固體、以水洗滌、繼而以己烷洗滌且在減壓下乾燥以提供290mg(85%)呈奶白色固體狀之5-胺基-1-(1-(4-氯苯基)乙基)-1H-1,2,3-***-4-甲酸。 To 5-Amino-1-(1-(4-chlorophenyl)ethyl)-1H-1,2,3-triazole-4-carboxylic acid ethyl ester (320 mg, 1.08 mmol) in ethanol (3 mL) A solution of NaOH (130 mg, 3.26 mmol) in water (2 mL) was added and the mixture was stirred at 60 &lt;0&gt;C for 4 h. The reaction mixture was concentrated under reduced pressure and the residue was crystallisjjjjjjjj The obtained solid was collected by filtration, washed with water, then washed with hexane and dried under reduced pressure to afford 290 mg (85%) of 5-amino-1-(1-(4-chlorobenzene ) as a creamy white solid. Ethyl)ethyl)-1H-1,2,3-triazole-4-carboxylic acid .

1H-NMR(400MHz,DMSO-d 6 )；δ 12.38(brs,1H),7.42(d,J=8.4Hz,2H),7.27(d,J=8.4Hz,2H),6.42(s,2H),5.71(q,J=7.2Hz,1H),1.83(d,J=7.2Hz,3H)；MS：m/z 267.0[M+H]+。 1 H-NMR (400 MHz, DMSO- d 6 ); δ 12.38 (brs, 1H), 7.42 (d, J = 8.4 Hz, 2H), 7.27 (d, J = 8.4 Hz, 2H), 6.42 (s, 2H) ), 5.71 (q, J = 7.2 Hz, 1H), 1.83 (d, J = 7.2 Hz, 3H); MS: m/z 267.0 [M+H] + .

步驟6：製備1-(1-(4-氯苯基)乙基)-1H-1,2,3-***-5-胺之兩種對映異構 體，E1及E2 Step 6: Preparation of two enantiomers of 1-(1-(4-chlorophenyl)ethyl)-1H-1,2,3-triazol-5-amine, E1 and E2

將5-胺基-1-(1-(4-氯苯基)乙基)-1H-1,2,3-***-4-甲酸(1.5g，5.62mmol)與N,N-二甲基苯胺(8mL)之混合物在200℃預熱油浴中攪拌15min。直接藉由使用石油醚中100% EtOAc之溶劑梯度作為溶離劑，經矽膠(100-200目)管柱層析純化經合併之反應團以提供850mg(34%)呈奶白色固體狀之1-(1-(4-氯苯基)乙基)-1H-1,2,3-***-5-胺，將其藉由對掌性製備型HPLC純化且分離兩種對映異構體E1與E2。 5-Amino-1-(1-(4-chlorophenyl)ethyl)-1H-1,2,3-triazole-4-carboxylic acid (1.5 g, 5.62 mmol) with N,N-dimethyl A mixture of the aniline (8 mL) was stirred in a 200 ° C preheated oil bath for 15 min. Directly by the use of 100% solvent gradient of EtOAc in petroleum ether as eluent, after silica gel (100-200 mesh) was purified by column chromatography were combined to provide a group of reaction 850mg (34%) of a white solid milk 1- (1-(4-Chlorophenyl)ethyl)-1H-1,2,3-triazol-5-amine , which was purified by preparative HPLC for separation and separation of two enantiomers E1 With E2 .

(S)-1-(1-(4-氯苯基)乙基)-1H-1,2,3-***-5-胺(E1) 1H-NMR(400MHz,DMSO-d 6 )；δ 7.39(d,J=8.4Hz,2H),7.23(d,J=8.4Hz,2H),6.8(s,1H),5.60(q,J=6.8Hz,1H),5.47(s,2H),1.81(d,J=6.8Hz,3H)；LCMS(ESI)：m/z 223.1[M+H]+；對掌性HPLC滯留時間：10.54min (S)-1-(1-(4-Chlorophenyl)ethyl)-1H-1,2,3-triazol-5-amine (E1) 1 H-NMR (400 MHz, DMSO- d 6 ); δ 7.39 (d, J = 8.4 Hz, 2H), 7.23 (d, J = 8.4 Hz, 2H), 6.8 (s, 1H), 5.60 (q, J = 6.8 Hz, 1H), 5.47 (s, 2H) , 1.81 (d, J = 6.8 Hz, 3H); LCMS (ESI): m/z 223.1 [M+H] + ;

(R)-1-(1-(4-氯苯基)乙基)-1H-1,2,3-***-5-胺(E2) 1H-NMR(400MHz,DMSO-d 6 )；δ 7.39(d,J=8.4Hz,2H),7.23(d,J=8.4Hz,2H),6.8(s,1H),5.60(q,J=6.8Hz,1H),5.47(s,2H),1.81(d,J=6.8Hz,3H)；LCMS(ESI)：m/z 223.1[M+H]+；對掌性HPLC滯留時間：12.37 min (R)-1-(1-(4-Chlorophenyl)ethyl)-1H-1,2,3-triazole-5-amine (E2) 1 H-NMR (400 MHz, DMSO- d 6 ); δ 7.39 (d, J = 8.4 Hz, 2H), 7.23 (d, J = 8.4 Hz, 2H), 6.8 (s, 1H), 5.60 (q, J = 6.8 Hz, 1H), 5.47 (s, 2H) , 1.81 (d, J = 6.8 Hz, 3H); LCMS (ESI): m/z 223.1 [M+H] + ;

步驟7：製備實例4-1及4-4Step 7: Prepare Examples 4-1 and 4-4

將1-(1-(4-氯苯基)乙基)-1H-1,2,3-***-5-胺 E1(1g，4.49mmol)、5-氯靛紅(0.816g，4.49mmol)及麥氏酸(0.647g，4.49mmol)於乙酸(30mL)中之混合物在100℃預熱油浴中攪拌4h。濃縮反應團且向殘餘物中添加水。藉由過濾收集所得固體，以水洗滌、繼而以己烷洗滌且在減壓下乾燥。藉由使用石油醚中35-50% EtOAc之溶劑梯度作為溶離劑，經矽膠(100-200目)管柱層析純化粗化合物(非對映異構體之混合物)以產生140mg(7%)呈奶白色固體狀之實例4-4，且以石油醚中60-100% EtOAc之溶劑梯度作為溶離劑純化產生220mg(11%)呈奶白色固體狀之實例4-1。 1-(1-(4-Chlorophenyl)ethyl)-1H-1,2,3-triazole-5-amine E1 (1 g, 4.49 mmol), 5-chloroindole (0.816 g, 4.49 mmol) And a mixture of the crude acid (0.647 g, 4.49 mmol) in acetic acid (30 mL) was stirred in a 100 ° C preheated oil bath for 4 h. The reaction mass was concentrated and water was added to the residue. The obtained solid was collected by filtration, washed with water, then washed with hexane and dried under reduced pressure. The crude compound (mixture of diastereomers) was purified by silica gel column chromatography (100-200 mesh) using a solvent gradient of 35-50% EtOAc in petroleum ether as solvent. Example 4-4, as a milky white solid, was purified eluting with EtOAc (EtOAc)

實例4-1之特徵資料展示如下： [α]25 D：-102.7(c=0.15,MeOH)。 The characteristics of Example 4-1 are shown below: [α] 25 D : -102.7 (c = 0.15, MeOH).

1H-NMR(400MHz,DMSO-d 6 )；δ 11.20(s,1H),10.69(s,1H),7.46(d,J=8.4Hz,2H),7.36(d,J=8.4Hz,2H),7.32(dd,J=2.4Hz,6.0Hz,2H),6.92(d,J=8.4Hz,1H),5.85(q,J=6.9Hz,1H),3.33(d,J=16.4Hz,1H),2.62(d,J=16.4Hz,1H),1.87(d,J=7.2Hz,3H)；LCMS(ESI)：m/z 427.9[M+H]+；r.t.=3.488；HPLC純度：在254nm下為97.82；對掌性HPLC純度：在254nm下為97.14。 1 H-NMR (400 MHz, DMSO- d 6 ); δ 11.20 (s, 1H), 10.69 (s, 1H), 7.46 (d, J = 8.4 Hz, 2H), 7.36 (d, J = 8.4 Hz, 2H) ), 7.32 (dd, J = 2.4 Hz, 6.0 Hz, 2H), 6.92 (d, J = 8.4 Hz, 1H), 5.85 (q, J = 6.9 Hz, 1H), 3.33 (d, J = 16.4 Hz, 1H), 2.62 (d, J = 16.4 Hz, 1H), 1.87 (d, J = 7.2 Hz, 3H); LCMS (ESI): m/z 427.9 [M+H] + rt=3.488; HPLC purity: 97.82 at 254 nm; palmitic HPLC purity: 97.14 at 254 nm.

實例4-4之特徵資料展示如下： [α]25 D：+31.2(c=0.16,MeOH)。 The characteristics of Example 4-4 are shown below: [α] 25 D : +31.2 (c = 0.16, MeOH).

1H-NMR(400MHz,DMSO-d 6 )；δ 11.29(s,1H),10.73(s,1H),7.46(d,J=8.8Hz,2H),7.38(d,J=8.8Hz,2H),7.32(dd,J=2.0Hz,7.2Hz,2H),6.93(d,J=8.8Hz,1H),5.80(q,J=6.8Hz,1H),3.23(d,J=16.4Hz,1H),2.73(d,J=16.0Hz,1H),1.87(d,J=7.2Hz,3H)；LCMS(ESI)：m/z 426[M-H]-；r.t.=1.94；HPLC純度：在254nm下為95.98。 1 H-NMR (400MHz, DMSO- d 6); δ 11.29 (s, 1H), 10.73 (s, 1H), 7.46 (d, J = 8.8Hz, 2H), 7.38 (d, J = 8.8Hz, 2H ), 7.32 (dd, J = 2.0 Hz, 7.2 Hz, 2H), 6.93 (d, J = 8.8 Hz, 1H), 5.80 (q, J = 6.8 Hz, 1H), 3.23 (d, J = 16.4 Hz, 1H), 2.73 (d, J = 16.0 Hz, 1H), 1.87 (d, J = 7.2 Hz, 3H); LCMS (ESI): m/z 426 [MH] - ; rt = 1.94; HPLC purity: 254 nm The next is 95.98.

使用類似於步驟7之程序，合成且分離實例4-2與實例4-3。 Example 4-2 and Example 4-3 were synthesized and separated using a procedure similar to that of Step 7.

實例4-2之特徵資料展示如下： [α]25 D：-24.0(c=0.30,MeOH)。 The characteristics of Example 4-2 are shown below: [α] 25 D : -24.0 (c = 0.30, MeOH).

1H NMR(400MHz,DMSO-d6)：δ ppm 1.86(d,J=6.8Hz,3H),2.72(d,J=16.4Hz,1H),3.21(d,J=16.4Hz,1H),5.80(q,J=6.8Hz,1H),6.93(d,J=9.2Hz,1H),7.33-7.31(m,2H),7.38(d,J=8.4Hz,2H),7.45(d,J=8.4Hz,2H),10.71(s,1H),11.27(s,1H)。MS：m/z 428.1(M+H)+。 1 H NMR (400MHz, DMSO- d 6): δ ppm 1.86 (d, J = 6.8Hz, 3H), 2.72 (d, J = 16.4Hz, 1H), 3.21 (d, J = 16.4Hz, 1H), 5.80 (q, J = 6.8 Hz, 1H), 6.93 (d, J = 9.2 Hz, 1H), 7.33 - 7.31 (m, 2H), 7.38 (d, J = 8.4 Hz, 2H), 7.45 (d, J = 8.4 Hz, 2H), 10.71 (s, 1H), 11.27 (s, 1H). MS: m/z 428.1 (M + H) + .

實例4-3之特徵資料展示如下： [α]25 D：+102.7(c=0.30,MeOH)。 The characteristics of Example 4-3 are shown below: [α] 25 D : +102.7 (c = 0.30, MeOH).

1H-NMR(400MHz,DMSO-d 6 )；δ 11.2(s,1H),10.69(s,1H),7.46(d,J=8.4Hz,2H),7.36(d,J=8.4Hz,2H),7.32(dd,J=2.4Hz,6.0Hz,2H),6.92(d,J=8.4Hz,1H),5.85(q,J=6.9Hz,1H),3.33(d,J= 16.4Hz,1H),2.62(d,J=16.4Hz,1H),1.87(d,J=7.2Hz,3H)；LCMS(ESI)：m/z 427.9[M+H]+ 1 H-NMR (400 MHz, DMSO- d 6 ); δ 11.2 (s, 1H), 10.69 (s, 1H), 7.46 (d, J = 8.4 Hz, 2H), 7.36 (d, J = 8.4 Hz, 2H) ), 7.32 (dd, J = 2.4 Hz, 6.0 Hz, 2H), 6.92 (d, J = 8.4 Hz, 1H), 5.85 (q, J = 6.9 Hz, 1H), 3.33 (d, J = 16.4 Hz, 1H), 2.62 (d, J = 16.4 Hz, 1H), 1.87 (d, J = 7.2 Hz, 3H); LCMS (ESI): m/z 427.9 [M+H] +

實例5Example 5 製備(R)-5'-氯-3-((5-氯吡啶-2-基)甲基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮 Preparation of (R)-5'-chloro-3-((5-chloropyridin-2-yl)methyl)-4,6-dihydrospiro[[1,2,3]triazolo[4,5- b]pyridine-7,3'-carboline]-2',5(3H)-dione

步驟1：製備(5-氯吡啶-2-基)甲醇 Step 1: Preparation of (5-chloropyridin-2-yl)methanol

在0℃下，向5-氯吡啶甲酸(5g，31.7mmol)於四氫呋喃(50mL)中之溶液中添加硼烷-二甲硫複合物(10mL)且將所得反應混合物在室溫下維持16h。將反應團冷卻至0℃，以甲醇(15mL)中止過量硼烷-二甲硫複合物，回流1h，濃縮反應，在乙酸乙酯(150mL)中稀釋，以水(2×50mL)、鹽水(50mL)洗滌，經無水硫酸鈉乾燥且濃縮以提供3g(66.6%產率)呈白色固體狀之(5-氯吡啶-2-基)甲醇。 To a solution of 5-chloropicolinic acid (5 g, 31.7 mmol) in tetrahydrofuran (50 mL) was added borane-dimethylsulfide complex (10 mL) and the obtained mixture was maintained at room temperature for 16 h. The reaction mixture was cooled to 0 ° C, and the excess borane-dimethylsulfide complex was quenched with methanol (15 mL), refluxed for 1 h, concentrated, and diluted in ethyl acetate (150 mL), water (2×50 mL), brine After washing with 50 mL), EtOAc (EtOAc m.

1H NMR(400MHz,CDCl3)：δ ppm 3.75(br.s,1H),4.75(s,2H),7.25(d,J=8.8Hz,1H),7.67(dd,J=2.2、6.1Hz 1H),8.53(d,J=8.8Hz,1H)。 1 H NMR (400 MHz, CDCl 3 ): δ ppm 3.75 (br.s, 1H), 4.75 (s, 2H), 7.25 (d, J = 8.8 Hz, 1H), 7.67 (dd, J = 2.2, 6.1 Hz 1H), 8.53 (d, J = 8.8 Hz, 1H).

步驟2：製備2-(溴甲基)-5-氯吡啶 Step 2: Preparation of 2-(bromomethyl)-5-chloropyridine

在0℃下，向(5-氯吡啶-2-基)甲醇(1.5g，10.45mmol)於***(20mL)中之溶液中添加三溴膦(2.83g，10.45mmol)於***中之溶液。將所得反應混合物在室溫下攪拌16h。將反應混合物傾入冰冷卻之水(50mL)中，使用固體碳酸氫鈉鹼化至ph-7，在***(2×75mL)中萃取，合併有機層，以水(2×70mL)、鹽水(30mL)洗滌，經無水Na2SO4乾燥且濃縮以提供1.6g(76%產率)呈粉色液體狀之2-(溴甲基)-5-氯吡啶。 A solution of tribromophosphine (2.83 g, 10.45 mmol) in diethyl ether was added to a solution of (5-chloropyridin-2-yl)methanol (1.5 g, 10.45 mmol) in diethyl ether (20 mL). The resulting reaction mixture was stirred at room temperature for 16 h. The reaction mixture was poured into EtOAc (EtOAc) (EtOAc) 30 mL), dried over anhydrous Na 2 SO 4 dried and concentrated to afford 1.6g (76% yield) as a pink liquid of 2- (bromomethyl) -5-chloropyridine.

1H NMR(400MHz,CDCl3)：δ ppm 4.52(s,2H),7.40(d,J=8.4Hz,1H),7.67(dd,J=2.2,6.1Hz 1H),8.53(d,J=2.7Hz,1H)。 1 H NMR (400MHz, CDCl 3 ): δ ppm 4.52 (s, 2H), 7.40 (d, J = 8.4Hz, 1H), 7.67 (dd, J = 2.2,6.1Hz 1H), 8.53 (d, J = 2.7 Hz, 1H).

步驟3：製備2-(疊氮基甲基)-5-氯吡啶 Step 3: Preparation of 2-(azidomethyl)-5-chloropyridine

在室溫下，向2-(溴甲基)-5-氯吡啶(750mg，3.64mmol)於二甲亞碸(8mL)中之溶液中添加疊氮化鈉(590mg，9.10mmol)且在相同溫度下攪拌2h。將反應混合物以***(50mL)稀釋，以水(2×40mL)、鹽水(30mL)洗滌，經無水Na2SO4乾燥且濃縮以提供0.5g(82%產率)呈無色液體狀之2-(疊氮基甲基)-5-氯吡啶。 Add sodium azide (590 mg, 9.10 mmol) to a solution of 2-(bromomethyl)-5-chloropyridine (750 mg, 3.64 mmol) in dimethyl hydrazine (8 mL) at rt. Stir at temperature for 2 h. The reaction mixture was diluted with diethyl ether (50mL), washed with water (2 × 40mL), brine (30mL), dried over anhydrous Na 2 SO 4 and concentrated to provide 0.5g (82% yield) as a colorless liquid of 2- (azidomethyl)-5-chloropyridine.

1H NMR(400MHz,CDCl3)：δ ppm 4.48(s,2H),7.31(d,J=8.3Hz,1H),7.71(dd,J=2.2,6.2Hz,1H),8.56(d,J=2.6Hz,1H)。 1 H NMR (400 MHz, CDCl 3 ): δ ppm 4.48 (s, 2H), 7.31 (d, J = 8.3 Hz, 1H), 7.71 (dd, J = 2.2, 6.2 Hz, 1H), 8.56 (d, J =2.6 Hz, 1H).

步驟4：製備5-胺基-1-((5-氯吡啶-2-基)甲基)-1H-1,2,3-***-4-甲酸乙酯 Step 4: Preparation of ethyl 5-amino-1-((5-chloropyridin-2-yl)methyl)-1H-1,2,3-triazole-4-carboxylate

在室溫下，向2-(疊氮基甲基)-5-氯吡啶(1.3g，7.71mmol)於乙醇中之溶液中添加氰基乙酸乙酯(1.31g，11.57mmol)及乙氧化鈉溶液(6.6mL，17.71mmol)。將所得反應混合物在80℃下維持4h。將反應混合物冷卻至室溫、濃縮、於乙酸乙酯(150mL)中稀釋、以水(2×50mL)及鹽水(50mL)洗滌、經無水Na2SO4乾燥且濃縮。藉由使用石油醚中30%乙酸乙酯之溶劑梯度作為溶離劑，經矽膠(100-200目)管柱層析純化所得固體以提供600mg(77.9%產率)呈白色固體狀之5-胺基-1-((5-氯吡啶-2-基)甲基)-1H-1,2,3-***-4-甲酸乙酯。 To a solution of 2-(azidomethyl)-5-chloropyridine (1.3 g, 7.71 mmol) in ethanol, ethyl cyanoacetate (1.31 g, 11.57 mmol) and sodium sulphate Solution (6.6 mL, 17.71 mmol). The resulting reaction mixture was maintained at 80 ° C for 4 h. The reaction mixture was cooled to room temperature, concentrated, (150 mL) diluted in ethyl acetate, washed with water (2 × 50mL) and brine (50mL), dried over anhydrous Na 2 SO 4 dried and concentrated. The resulting solid was purified by silica gel (100-200 mesh) column chromatography eluting eluting eluting eluting eluting Ethyl-1-((5-chloropyridin-2-yl)methyl)-1H-1,2,3-triazole-4-carboxylate.

1H NMR(400MHz,DMSO-d6)：δ ppm 1.24(t,J=8.0Hz,3H),4.26(q,J=6.8Hz,2H),5.56(s,2H),6.58(br s,2H),7.21(d,J=8.0Hz,1H),7.95(dd,J=2.4,6.0Hz,1H),8.58(d,J=1.7Hz,1H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm 1.24 (t, J = 8.0Hz, 3H), 4.26 (q, J = 6.8Hz, 2H), 5.56 (s, 2H), 6.58 (br s, 2H), 7.21 (d, J = 8.0 Hz, 1H), 7.95 (dd, J = 2.4, 6.0 Hz, 1H), 8.58 (d, J = 1.7 Hz, 1H).

步驟5：製備5-胺基-1-((5-氯吡啶-2-基)甲基)-1H-1,2,3-***-4-甲酸 Step 5: Preparation of 5-amino-1-((5-chloropyridin-2-yl)methyl)-1H-1,2,3-triazole-4-carboxylic acid

在室溫下，向5-胺基-1-((5-氯吡啶-2-基)甲基)-1H-1,2,3-***-4-甲酸乙酯(600mg，2.14mmol)於乙醇(3mL)中之溶液中添加水(3mL)，繼而添加氫氧化鈉(256mg，6.41mmol)且將所得反應混合物在60℃下維持3h。將所得反應混合物冷卻至室溫、濃縮以移除乙醇、使用飽和硫酸氫鈉酸化至pH 6，以水(2×15mL)洗滌經過濾獲得之固 體且在真空下乾燥以獲得350mg(64.8%產率)呈白色固體狀之5-胺基-1-((5-氯吡啶-2-基)甲基)-1H-1,2,3-***-4-甲酸。 To 5-Amino-1-((5-chloropyridin-2-yl)methyl)-1H-1,2,3-triazole-4-carboxylic acid ethyl ester (600 mg, 2.14 mmol) Water (3 mL) was added to a solution in EtOAc (3 mL). The resulting reaction mixture was cooled to room temperature, concentrated to remove ethanol, acidified to pH 6 using saturated sodium hydrogen sulfate, washed with water (2×15 mL) Drying under vacuum to give 350 mg (64.8% yield) of 5-amino-1-((5-chloropyridin-2-yl)methyl)-1H-1,2,3- Triazole-4-carboxylic acid.

1H NMR(400MHz,DMSO-d6)：5.55(s,2H),6.52(br s,2H),7.20(d,J=8.3Hz,1H),7.95(dd,J=1.8,6.6Hz,1H),8.59(d,J=2.2Hz,1H),12.48(s,1H)。 1 H NMR (400MHz, DMSO- d 6): 5.55 (s, 2H), 6.52 (br s, 2H), 7.20 (d, J = 8.3Hz, 1H), 7.95 (dd, J = 1.8,6.6Hz, 1H), 8.59 (d, J = 2.2 Hz, 1H), 12.48 (s, 1H).

MS：m/z 254.3(M+H)+。 MS: m/z 254.3 (M + H) + .

步驟6：製備1-((5-氯吡啶-2-基)甲基)-1H-1,2,3-***-5-胺 Step 6: Preparation of 1-((5-chloropyridin-2-yl)methyl)-1H-1,2,3-triazole-5-amine

將5-胺基-1-((5-氯吡啶-2-基)甲基)-1H-1,2,3-***-4-甲酸(300mg，1.19mmol)於N,N-二甲基苯胺(2mL)中之懸浮液在200℃下維持20min。將反應團冷卻至室溫且以石油醚洗滌以移除N,N-二甲基苯胺。藉由使用石油醚中50%乙酸乙酯之溶劑梯度作為溶離劑，經矽膠(100-200目)管柱層析純化所得固體以提供110mg(44%產率)呈白色固體狀之1-((5-氯吡啶-2-基)甲基)-1H-1,2,3-***-5-胺。 5-Amino-1-((5-chloropyridin-2-yl)methyl)-1H-1,2,3-triazole-4-carboxylic acid (300 mg, 1.19 mmol) in N , N -dimethyl The suspension in the aniline (2 mL) was maintained at 200 ° C for 20 min. The reaction mass was cooled to room temperature and washed with petroleum ether to remove N , N -dimethylaniline. The resulting solid was purified by silica gel (100-200 mesh) column chromatography eluting with EtOAc (EtOAc) elute (5-Chloropyridin-2-yl)methyl)-1H-1,2,3-triazole-5-amine.

1H NMR(400MHz,DMSO-d6)：5.46(s,2H),5.62(br s,2H),6.85(s,1H),7.03(d,J=8.8Hz,1H),7.94(dd,J=2.2,6.1Hz,1H),8.6(d,J=2.7Hz,1H)。 1 H NMR (400MHz, DMSO- d 6): 5.46 (s, 2H), 5.62 (br s, 2H), 6.85 (s, 1H), 7.03 (d, J = 8.8Hz, 1H), 7.94 (dd, J = 2.2, 6.1 Hz, 1H), 8.6 (d, J = 2.7 Hz, 1H).

MS：m/z 210.3(M+H)+。 MS: m/z 210.3 (M + H) + .

步驟7：製備5'-氯-3-((5-氯吡啶-2-基)甲基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮 Step 7: Preparation of 5'-chloro-3-((5-chloropyridin-2-yl)methyl)-4,6-dihydrospiro[[1,2,3]triazolo[4,5-b Pyridine-7,3'-carboline]-2',5(3H)-dione

在室溫下，向1-((5-氯吡啶-2-基)甲基)-1H-1,2,3-***-5-胺(110mg，0.53mmol)於乙酸中之溶液中添加5-氯吲哚啉-2,3-二酮(95.6mg，0.53mmol)、麥氏酸(75.7mg，0.53mmol)且在100℃下維持2h。將反應混合物冷卻至室溫且在減壓下移除溶劑。向反應混合物中添加冰水且在室溫下攪拌15min。以水(2×5mL)洗滌經過濾獲得之固體且在真空下乾燥。藉由使用DCM中10% MeOH之溶劑梯度作為溶離劑，經矽膠(100-200目)管柱層析純化所得固體以提供32mg(16%產率)呈淺黃色固體狀之5'-氯-3-((5-氯吡啶-2-基)甲基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮。 Add to a solution of 1-((5-chloropyridin-2-yl)methyl)-1H-1,2,3-triazole-5-amine (110 mg, 0.53 mmol) in acetic acid at room temperature 5-Chloroporphyrin-2,3-dione (95.6 mg, 0.53 mmol), Malt's acid (75.7 mg, 0.53 mmol) and maintained at 100 ° C for 2 h. The reaction mixture was cooled to room temperature and the solvent was removed under reduced pressure. Ice water was added to the reaction mixture and stirred at room temperature for 15 min. The solid obtained by filtration was washed with water (2 x 5 mL) and dried under vacuum. The resulting solid was purified by silica gel (100-200 mesh) column chromatography eluting eluting eluting eluting eluting 3-((5-chloropyridin-2-yl)methyl)-4,6-dihydrospiro[[1,2,3]triazolo[4,5-b]pyridine-7,3'-oxime Porphyrin]-2',5(3H)-dione.

1H NMR(400MHz,DMSO-d6)：δ ppm 2.73(d,J=16.3Hz,1H),3.30(d,J=16.3Hz,1H),5.66(s,2H),6.94(d,J=8.0Hz,1H),7.30-7.40(m,3H),8.00(dd,J=2.7Hz and 5.7Hz,1H),8.63(d,J=2.2Hz,1H),10.77(s,1H),11.29(br s,1H)。 1 H NMR (400 MHz, DMSO- d6 ): δ ppm 2.73 (d, J = 16.3 Hz, 1H), 3.30 (d, J = 16.3 Hz, 1H), 5.66 (s, 2H), 6.94 (d, J = 8.0 Hz, 1H), 7.30-7.40 (m, 3H), 8.00 (dd, J = 2.7 Hz and 5.7 Hz, 1H), 8.63 (d, J = 2.2 Hz, 1H), 10.77 (s, 1H), 11.29 (br s, 1H).

MS：m/z 415.0(M+H)+。 MS: m/z 415.0 (M + H) + .

步驟8：製備(R)-5'-氯-3-((5-氯吡啶-2-基)甲基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮 Step 8: Preparation of (R)-5'-chloro-3-((5-chloropyridin-2-yl)methyl)-4,6-dihydrospiro[[1,2,3]triazolo[4 ,5-b]pyridine-7,3'-carboline]-2',5(3H)-dione

可藉由使用上述實例之程序(例如，對掌性製備型HPLC)分離5'-氯-3-((5-氯吡啶-2-基)甲基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮來製備實例5，(R)-5'-氯-3-((5-氯吡啶-2-基)甲基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮。 5'-Chloro-3-((5-chloropyridin-2-yl)methyl)-4,6-dihydrospiro[[] can be isolated by using the procedure of the above examples (for example, for preparative HPLC). 1,2,3] Triazolo[4,5-b]pyridine-7,3'-carboline]-2',5(3H)-dione to prepare Example 5, (R)-5'- Chloro-3-((5-chloropyridin-2-yl)methyl)-4,6-dihydrospiro[[1,2,3]triazolo[4,5-b]pyridine-7,3' -Porphyrin]-2',5(3H)-dione.

實例6Example 6 製備(R)-5'-氯-3-(4-氯苄基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮 Preparation of (R)-5'-chloro-3-(4-chlorobenzyl)-4,6-dihydrospiro[[1,2,3]triazolo[4,5-b]pyridine-7,3 '-Porphyrin-2',5(3H)-dione

步驟1：製備1-(疊氮基甲基)-4-氯苯 Step 1: Preparation of 1-(azidomethyl)-4-chlorobenzene

在室溫下，向1-氯-4-(氯甲基)苯(5g，31.05mmol)於二甲亞碸(40mL)中之溶液中添加疊氮化鈉(5.06g，77.63mmol)且在相同溫度下攪拌5h。將反應混合物以乙酸乙酯(100mL)稀釋，以水(2×70mL)、鹽水(30mL)洗滌，經無水硫酸鈉乾燥且濃縮以提供4.5g(86%產率)呈無色液體狀之1-(疊氮基甲基)-4-氯苯。 Add sodium azide (5.06 g, 77.63 mmol) to a solution of 1-chloro-4-(chloromethyl)benzene (5 g, 31.05 mmol) in dimethylhydrazine (40 mL) at rt. Stir at the same temperature for 5 h. The reaction mixture was diluted with ethyl acetate (100 mL), water (2 × 70mL), brine (30 mL), dried over anhydrous sodium sulfate and concentrated to afford 4.5g (86% yield) as a colorless liquid of 1- (azidomethyl)-4-chlorobenzene .

1H NMR(400MHz,CDCl3)：δ ppm 4.32(s,2H),7.25-7.26(d,2H,與CDCl3合併)，7.36(d,J=8.4Hz,2H)。 1 H NMR (400 MHz, CDCl 3 ): δ ppm 4.32 (s, 2H), 7.25-7.26 (d, 2H, combined with CDCl 3 ), 7.36 (d, J = 8.4 Hz, 2H).

步驟2：製備5-胺基-1-(4-氯苄基)-1H-1,2,3-***-4-甲酸 Step 2: Preparation of 5-amino-1-(4-chlorobenzyl)-1H-1,2,3-triazole-4-carboxylic acid

在室溫下，向鈉(550mg，23.88mmol)於乙醇中之溶液中添加2-氰基乙酸(1.01g，11.94mmol)及1-(疊氮基甲基)-4-氯苯(2g，11.94mmol)。將所得反應混合物在80℃下維持16h。將反應混合物冷卻至0℃；向反應混合物中添加冰水且在室溫下攪拌15min、以HCl水溶液酸化至pH 1。將經過濾獲得之固體以水(2×15mL)、***(2×10mL)洗滌且在真空下乾燥以提供720mg(25%產率)呈白色固體狀之5-胺基-1-(4-氯苄基)-1H-1,2,3-***-4-甲酸。 To a solution of sodium (550 mg, 23.88 mmol) in ethanol was added 2-cyanoacetic acid (1.01 g, 11.94 mmol) and 1-(azidomethyl)-4-chlorobenzene (2 g, 11.94 mmol). The resulting reaction mixture was maintained at 80 ° C for 16 h. The reaction mixture was cooled to 0 ° C; ice water was added and the mixture was stirred at room temperature for 15 min. The solid was filtered to obtain the water (2 × 15mL), ether (2 × 10mL), and washed and dried in vacuo to provide 720mg (25% yield) as a white solid of 5-amino-1- (4 Chlorobenzyl)-1H-1,2,3-triazole-4-carboxylic acid .

1H NMR(400MHz,DMSO-d6)：δ ppm 5.42(s,2H),6.55(br s,2H),7.22(d,J=7.5Hz,2H),7.43(d,J=7.1Hz,2H),12.49(br s,1H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm 5.42 (s, 2H), 6.55 (br s, 2H), 7.22 (d, J = 7.5Hz, 2H), 7.43 (d, J = 7.1Hz, 2H), 12.49 (br s, 1H).

MS：m/z 253.3(M+H)+。 MS: m/z 253.3 (M + H) + .

步驟3：製備1-(4-氯苄基)-1H-1,2,3-***-5-胺 Step 3: Preparation of 1-(4-chlorobenzyl)-1H-1,2,3-triazole-5-amine

將5-胺基-1-(4-氯苄基)-1H-1,2,3-***-4-甲酸(500mg，1.98mmol)於N,N-二甲基苯胺中之懸浮液在230℃下維持30min。將反應團冷卻至室溫且以石油醚洗滌以移除N,N-二甲基苯胺。藉由使用石油醚 中50%乙酸乙酯之溶劑梯度作為溶離劑，經矽膠(100-200目)管柱層析純化所得固體以提供130mg(31%產率)呈白色固體狀之1-(4-氯苄基)-1H-1,2,3-***-5-胺。 A suspension of 5-amino-1-(4-chlorobenzyl)-1H-1,2,3-triazole-4-carboxylic acid (500 mg, 1.98 mmol) in N , N -dimethylaniline Maintain at 230 ° C for 30 min. The reaction mass was cooled to room temperature and washed with petroleum ether to remove N , N -dimethylaniline. By using a solvent gradient of 50% ethyl acetate in petroleum ether as the eluent, dried over silica gel (100-200 mesh) The resulting solid was purified by column chromatography to provide 130mg (31% yield) as a white solid of 1- ( 4-chlorobenzyl)-1H-1,2,3-triazole-5-amine .

1H NMR(400MHz,DMSO-d6)：δ ppm 5.32(s,2H),6.16(br s,2H),7.20(d,J=8.3Hz,2H),7.41(d,J=8.3Hz,2H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm 5.32 (s, 2H), 6.16 (br s, 2H), 7.20 (d, J = 8.3Hz, 2H), 7.41 (d, J = 8.3Hz, 2H).

步驟4：製備5'-氯-3-(4-氯苄基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮 Step 4: Preparation of 5'-chloro-3-(4-chlorobenzyl)-4,6-dihydrospiro[[1,2,3]triazolo[4,5-b]pyridine-7,3' -porphyrin]-2',5(3H)-dione

在室溫下，向1-(4-氯苄基)-1H-1,2,3-***-5-胺(130mg，0.625mmol)於乙酸中之溶液中添加5-氯吲哚啉-2,3-二酮(113mg，0.625mmol)、麥氏酸(90mg，0.625mmol)且在100℃下維持2h。將反應混合物冷卻至室溫且在減壓下移除溶劑。向反應混合物中添加冰水且在室溫下攪拌15min。將經過濾獲得之固體以水(2×5mL)洗滌且在真空下乾燥。以丙酮洗滌所得固體以提供80mg(31%產率)呈淺黃色固體狀之5'-氯-3-(4-氯苄基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮。 Add 5-chloroporphyrin to a solution of 1-(4-chlorobenzyl)-1H-1,2,3-triazole-5-amine (130 mg, 0.625 mmol) in acetic acid at room temperature. 2,3-Dione (113 mg, 0.625 mmol), Malt's acid (90 mg, 0.625 mmol) and maintained at 100 ° C for 2 h. The reaction mixture was cooled to room temperature and the solvent was removed under reduced pressure. Ice water was added to the reaction mixture and stirred at room temperature for 15 min. The solid obtained by filtration was washed with water (2×5 mL) and dried under vacuum. The resulting solid was washed with EtOAc (EtOAc) (EtOAc:EtOAc : Triazolo[4,5-b]pyridine-7,3'-carboline]-2 ' ,5(3H)-dione .

1H-NMR(DMSO-d6)：1H NMR(400MHz,DMSO-d6)：δ ppm 2.68(d,J=16.3Hz,1H),3.30(d，與水分合併，1H),5.53(s,2H),6.93(d,J=8.4Hz,1H),7.30-7.42(m,4H),7.48(d,J=8.3Hz,2H),10.75(s,1H),11.38(s,1H)。 1 H-NMR (DMSO-d 6 ): 1 H NMR (400 MHz, DMSO- d6 ): δ ppm 2.68 (d, J = 16.3 Hz, 1H), 3.30 (d, combined with water, 1H), 5.53 (s) , 2H), 6.93 (d, J = 8.4 Hz, 1H), 7.30-7.42 (m, 4H), 7.48 (d, J = 8.3 Hz, 2H), 10.75 (s, 1H), 11.38 (s, 1H) .

LC/MS：m/z：414.0(M+H)+。 LC / MS: m / z: 414.0 (M + H) +.

步驟5：製備(R)-5'-氯-3-(4-氯苄基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮 Step 5: Preparation of (R)-5'-chloro-3-(4-chlorobenzyl)-4,6-dihydrospiro[[1,2,3]triazolo[4,5-b]pyridine- 7,3'-porphyrin]-2',5(3H)-dione

可藉由使用上述實例之程序(例如，對掌性製備型HPLC)分離5'-氯-3-(4-氯苄基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮來製備實例6，(R)-5'-氯-3-(4-氯苄基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮。 5'-Chloro-3-(4-chlorobenzyl)-4,6-dihydrospiro[[1,2,3]3 can be isolated by using the procedure of the above examples (for example, for preparative HPLC) Preparation of oxazo[4,5-b]pyridine-7,3'-carboline]-2',5(3H)-dione to give Example 6 , (R)-5'-chloro-3-(4- Chlorobenzyl)-4,6-dihydrospiro[[1,2,3]triazolo[4,5-b]pyridine-7,3'-carboline]-2',5(3H)- Dione.

使用通用程序以及上述實例之程序，由適當起始物質來製備以下化合物。 The following compounds were prepared from the appropriate starting materials using the general procedures and procedures of the above examples.

實例7-1Example 7-1 (R)-5-氯-1'-(4-氯-2-氟苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-chloro-1'-(4-chloro-2-fluorobenzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4- b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為5-氯-1'-(4-氯-2-氟苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H NMR(400MHz,DMSO-d6)：δ ppm 2.53(d,J=20.7Hz,1H),3.12(d,J=15.9Hz,1H),5.33(q,J=15.9Hz,2H),6.91(d,J=7.8Hz 1H),6.80(s,1H),7.07(t,J=8.4Hz 1H),7.27-7.33(m,3H),7.47(dd,J=1.7,1.7Hz,1H),10.6(s,1H),11.08(br s,1H)。 The above-mentioned title compound racemate, which is 5-chloro-1 '-(4-chloro-2-fluorobenzyl)-5',7'-dihydrospiro, is prepared using the general procedure and the procedure of the above example. Porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristics are: 1 H NMR (400 MHz, DMSO-d 6 ): δ ppm 2.53 (d, J = 20.7 Hz, 1H), 3.12 (d, J = 15.9 Hz, 1H), 5.33 (q, J = 15.9 Hz, 2H), 6.91 (d, J = 7.8 Hz 1H) , 6.80 (s, 1H), 7.07 (t, J = 8.4 Hz 1H), 7.27-7.33 (m, 3H), 7.47 (dd, J = 1.7, 1.7 Hz, 1H), 10.6 (s, 1H), 11.08 (br s, 1H).

MS：m/z 431.0(M+H)+。 MS: m/z 431.0 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-2Example 7-2 (R)-5-氯-1'-(4-(三氟甲氧基)苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-chloro-1'-(4-(trifluoromethoxy)benzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3, 4-b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為5-氯-1'-(4-(三氟甲氧基)苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H NMR(400MHz,DMSO-d6)：δ ppm 2.54(d,J=16.5Hz,1H),3.15(d,J=16.3Hz,1H),5.35(ABq,J=15.8Hz,2H),6.80(s,1H),6.91(d,J=7.9Hz,1H),7.35(m,6H),10.60(s,1H),11.1(br s,1H)。 The above-mentioned title compound racemate, which is 5-chloro-1 '-(4-(trifluoromethoxy)benzyl)-5',7'-dihydrogen, was prepared using the general procedure and procedure of the above example. Spirulina [porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristics are: 1 H NMR (400 MHz, DMSO- D6 ): δ ppm 2.54 (d, J = 16.5 Hz, 1H), 3.15 (d, J = 16.3 Hz, 1H), 5.35 (ABq, J = 15.8 Hz, 2H), 6.80 (s, 1H), 6.91 ( d, J = 7.9 Hz, 1H), 7.35 (m, 6H), 10.60 (s, 1H), 11.1 (br s, 1H).

MS：m/z 463.1(M+H)+。 MS: m/z 463.1 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-3Example 7-3 (R)-5-氯-1'-(4-甲氧基苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-chloro-1'-(4-methoxybenzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b] Pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為5-氯-1'-(4-甲氧基苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H NMR(400MHz,DMSO-d6)：δ ppm 2.56(s,1H，合併於DMSO中)，3.08(d,J=16.2Hz,1H),3.73(s,3H),5.20(ABq,J=15.0Hz,2H),6.74(s,1H),6.90(d,J=6.6Hz,1H),7.2-7.3(m,4H),10.58(br,s,1H),11.08(br,s,1H)。 The above-mentioned title compound racemate, which is 5-chloro-1 '-(4-methoxybenzyl)-5',7'-dihydrospiro[吲哚, is prepared using the general procedure and procedures of the above examples. -3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristics are: 1 H NMR (400 MHz, DMSO- d6 ): δ Ppm 2.56 (s, 1H, combined in DMSO), 3.08 (d, J = 16.2 Hz, 1H), 3.73 (s, 3H), 5.20 (ABq, J = 15.0 Hz, 2H), 6.74 (s, 1H) , 6.90 (d, J = 6.6 Hz, 1H), 7.2-7.3 (m, 4H), 10.58 (br, s, 1H), 11.08 (br, s, 1H).

MS：m/z 407.1(M-H)-。 MS: m/z 407.1 (MH) - .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-4Example 7-4 (R)-5-氯-1'-(4-(二氟甲氧基)苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-chloro-1'-(4-(difluoromethoxy)benzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3, 4-b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為5-氯-1'-(4-(二氟甲氧基)苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為： 1H-NMR(400MHz,DMSO-d6)：δ ppm 2.53(d,1H，與DMSO合併)，3.13(d,J=16.3Hz,1H),5.27(ABq,J=15.8Hz,2H),6.78(s,1H),6.91(d,J=8.30Hz,1H),7.16(d,J=8.4Hz,2H),7.21(t,J=74.2Hz,1H),7.28-7.34(m,4H),10.60(s,1H),11.10(br s,1H)。 The above-mentioned title compound racemate, which is 5-chloro-1 '-(4-(difluoromethoxy)benzyl)-5',7'-dihydrogen, was prepared using the general procedure and procedure of the above example. Spiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristics are: 1 H-NMR (400MHz, DMSO -d 6 ): δ ppm 2.53 (d, 1H, combined with DMSO), 3.13 (d, J = 16.3 Hz, 1H), 5.27 (ABq, J = 15.8 Hz, 2H), 6.78 (s, 1H), 6.91 (d, J = 8.30 Hz, 1H), 7.16 (d, J = 8.4 Hz, 2H), 7.21 (t, J = 74.2 Hz, 1H), 7.28-7.34 (m, 4H), 10.60 (s, 1H) , 11.10 (br s, 1H).

MS：m/z 445.1(M+H)+。 MS: m/z 445.1 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-5Example 7-5 (R)-5-氯-1'-(2,4-二甲氧基苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-chloro-1'-(2,4-dimethoxybenzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4 -b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為5-氯-1'-(2,4-二甲氧基苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H NMR(400MHz,DMSO-d 6)δ ppm 2.56(d,J=15.97Hz,1 H),3.11(d,J=15.97Hz,1 H),3.63(s,3 H),3.76(s,3 H),5.15-5.27(m,2 H),6.23(d,J=3.00Hz,1 H),6.79(s,1 H),6.82(dd,J=3.08,3.04Hz,1 H),6.93(t,J=9.10,2 H),7.29(s,1 H),7.31(d,J=2.16Hz,1 H),10.62(s,1 H),10.95(s,1 H)。 The above-mentioned title compound racemate, which is 5-chloro-1 '-(2,4-dimethoxybenzyl)-5',7'-dihydrospiro, was prepared using the general procedure and procedures of the above examples. [Porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristics are: 1 H NMR (400 MHz, DMSO- d 6 ) δ ppm 2.56 (d, J = 15.97 Hz, 1 H), 3.11 (d, J = 15.97 Hz, 1 H), 3.63 (s, 3 H), 3.76 (s, 3 H), 5.15-5.27 ( m, 2 H), 6.23 (d, J = 3.00 Hz, 1 H), 6.79 (s, 1 H), 6.82 (dd, J = 3.08, 3.04 Hz, 1 H), 6.93 (t, J = 9.10, 2 H), 7.29 (s, 1 H), 7.31 (d, J = 2.16 Hz, 1 H), 10.62 (s, 1 H), 10.95 (s, 1 H).

ES-MS：439.1(M+H)+。 ES-MS: 439.1 (M+H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-6Example 7-6 (R)-1'-(4-氯苄基)-5-甲氧基-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-1'-(4-chlorobenzyl)-5-methoxy-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b] Pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為1'-(4-氯苄基)-5-甲氧基-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H NMR(400MHz,DMSO-d6)：δ ppm2.50(d，質子與DMSO合併，1H),3.06(d,J=15.8Hz,1H),5.27(ABq,J=15.8Hz,2H),5.30(d,J=15.8Hz,1H),6.75(s,1H),6.81(s,2H),6.86(s,1H),7.27(d,J=7.9Hz,2H),7.42(d,J=8.4Hz,2H),10.28(s,1H),11.05(br s,1H)。 The above-mentioned title compound racemate, which is 1'-(4-chlorobenzyl)-5-methoxy-5',7'-dihydrospiro[吲哚, is prepared using the general procedure and the procedure of the above example. -3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristics are: 1 H NMR (400 MHz, DMSO-d 6 ): δ ppm 2.50 (d, proton and DMSO combined, 1H), 3.06 (d, J = 15.8 Hz, 1H), 5.27 (ABq, J = 15.8 Hz, 2H), 5.30 (d, J = 15.8 Hz, 1H) , 6.75 (s, 1H), 6.81 (s, 2H), 6.86 (s, 1H), 7.27 (d, J = 7.9 Hz, 2H), 7.42 (d, J = 8.4 Hz, 2H), 10.28 (s, 1H), 11.05 (br s, 1H).

MS：m/z 409.1(M+H)+。 MS: m/z 409.1 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-7Example 7-7 (S)-5'-氯-1-(4-氯苄基)-5,7-二氫螺[吡唑并[3,4-b]吡啶-4,3'-吡咯并[2,3-b]吡啶]-2',6(1H,1'H)-二酮 (S)-5'-chloro-1-(4-chlorobenzyl)-5,7-dihydrospiro[pyrazolo[3,4-b]pyridine-4,3'-pyrrolo[2,3 -b]pyridine]-2',6(1H,1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為5'-氯-1-(4-氯苄基)-5,7-二氫螺[吡唑并[3,4-b]吡啶-4,3'-吡咯并[2,3-b]吡啶]-2',6(1H,1'H)-二酮，且特徵資料為：1H NMR(400MHz,DMSO-d6)：δ ppm 2.61(d,J=16.2Hz,1H)。3.21(d,J=15.8Hz,1H),5.28(ABq,J=15.8Hz,2H),6.91(s,1H),7.26(dd,J=8.4Hz,2H),7.43(dd,J=8.0Hz,2H),7.82(d,J=1.7Hz,1H),8.20(d,J=1.8Hz,1H),11.15(s,1H),11.31(br s,1H)。 The above-mentioned title compound racemate, which is 5'-chloro-1-(4-chlorobenzyl)-5,7-dihydrospiro[pyrazolo[3, 4-b]pyridine-4,3'-pyrrolo[2,3-b]pyridine]-2',6(1H,1'H)-dione, and the characteristics are: 1 H NMR (400 MHz, DMSO -d 6 ): δ ppm 2.61 (d, J = 16.2 Hz, 1H). 3.21 (d, J = 15.8 Hz, 1H), 5.28 (ABq, J = 15.8 Hz, 2H), 6.91 (s, 1H), 7.26 (dd, J = 8.4 Hz, 2H), 7.43 (dd, J = 8.0) Hz, 2H), 7.82 (d, J = 1.7 Hz, 1H), 8.20 (d, J = 1.8 Hz, 1H), 11.15 (s, 1H), 11.31 (br s, 1H).

MS：m/z 414.0(M+H)+。 MS: m/z 414.0 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-8Example 7-8 (R)-5-氯-1'-(4-氯苄基)-1-環丙基-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-chloro-1'-(4-chlorobenzyl)-1-cyclopropyl-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3, 4-b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為5-氯-1'-(4-氯苄基)-1-環丙基-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H NMR(400MHz,DMSO-d6)：δ ppm 0.77(d,J=18.0Hz,2H),0.97(d,J=6.2Hz,2H),2.55(d,1H，與DMSO合併)，2.67-2.70(m,1H),3.12(d,J=16.3Hz,1H),5.27(ABq,J=15.9Hz,2H),6.75(s,1H),7.18(d,J=8.4Hz,1H),7.26(d,J=8.4Hz,2H),7.36(d,J=1.6Hz,1H),7.42-7.44(m,3H),11.12(br s,1H)。 The above-mentioned title compound racemate, which is 5-chloro-1 '-(4-chlorobenzyl)-1-cyclopropyl-5',7'-dihydrogen, was prepared using the general procedure and procedure of the above example. Spirulina [porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristics are: 1 H NMR (400 MHz, DMSO- d 6 ): δ ppm 0.77 (d, J = 18.0 Hz, 2H), 0.97 (d, J = 6.2 Hz, 2H), 2.55 (d, 1H, combined with DMSO), 2.67-2.70 (m, 1H), 3.12 (d, J = 16.3 Hz, 1H), 5.27 (ABq, J = 15.9 Hz, 2H), 6.75 (s, 1H), 7.18 (d, J = 8.4 Hz, 1H), 7.26 (d, J = 8.4) Hz, 2H), 7.36 (d, J = 1.6 Hz, 1H), 7.42-7.44 (m, 3H), 11.12 (br s, 1H).

MS：m/z 453.0(M+H)+。 MS: m/z 453.0 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-9Example 7-9 (R)-1'-(4-氯苄基)-5',7'-二氫螺[[1,3]間二氧雜環戊烯并[4,5-f]吲哚-7,4'-吡唑并[3,4-b]吡啶]-6,6'(1'H,5H)-二酮 (R)-1'-(4-chlorobenzyl)-5',7'-dihydrospiro[[1,3]dioxole[4,5-f]indole-7, 4'-pyrazolo[3,4-b]pyridine]-6,6'(1'H,5H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為1'-(4-氯苄基)-5',7'-二氫螺[[1,3]間二氧雜環戊烯并[4,5-f]吲哚-7,4'-吡唑并[3,4-b]吡啶]-6,6'(1'H,5H)-二酮，且特徵資料為：1H NMR(400MHz,DMSO-d 6)d ppm 2.42(d,J=15.8Hz,1 H),3.05(d,J=15.8Hz,1 H),5.27(d,J=10.8Hz,2H),5.90-6.02(m,2H),6.55(s,1H),6.78(s,1H),6.88(s,1H),7.27(d,J=8.5Hz,2 H),7.38-7.48(m,2 H),10.26(s,1 H),11.03(s,1 H)。 The above-mentioned title compound racemate, which is 1'-(4-chlorobenzyl)-5',7'-dihydrospiro[[1,3]dioxane, was prepared using the general procedure and the procedure of the above example. Heterocyclopenta[4,5-f]indole-7,4'-pyrazolo[3,4-b]pyridine]-6,6'(1'H,5H)-dione, and characterized The data are: 1 H NMR (400 MHz, DMSO- d 6 ) d ppm 2.42 (d, J = 15.8 Hz, 1 H), 3.05 (d, J = 15.8 Hz, 1 H), 5.27 (d, J = 10.8 Hz) , 2H), 5.90-6.02 (m, 2H), 6.55 (s, 1H), 6.78 (s, 1H), 6.88 (s, 1H), 7.27 (d, J = 8.5 Hz, 2 H), 7.38-7.48 (m, 2 H), 10.26 (s, 1 H), 11.03 (s, 1 H).

MS：m/z 423.1(M+H)+。 MS: m/z 423.1 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-10Example 7-10 (R)-5-溴-1'-(4-氯苄基)-6-氟-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-bromo-1'-(4-chlorobenzyl)-6-fluoro-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4- b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為5-溴-6-氯-1'-(4-氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H NMR(400MHz,DMSO-d 6)δ ppm 2.52(d,J=15.97Hz,1H),3.15(d,J=15.97Hz,1H),5.22-5.33(m,2H),6.82(s,1H),6.92(d,J=8.92Hz,1H),7.26(d,J=8.44,2H),7.42(d,J=8.44、2H),7.57(d,J=6.88、1H),10.77(s,1H),11.04(s,1H)。 The above-mentioned title compound racemate, which is 5-bromo-6-chloro-1 '-(4-chlorobenzyl)-5',7'-dihydrospiro, was prepared using the general procedure and the procedure of the above example. Porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristics are: 1 H NMR (400 MHz, DMSO- d 6 ) δ ppm 2.52 (d, J = 15.97 Hz, 1H), 3.15 (d, J = 15.97 Hz, 1H), 5.22-5.33 (m, 2H), 6.82 (s, 1H), 6.92 (d, J = 8.92) Hz, 1H), 7.26 (d, J = 8.44, 2H), 7.42 (d, J = 8.44, 2H), 7.57 (d, J = 6.88, 1H), 10.77 (s, 1H), 11.04 (s, 1H) ).

ES-MS：m/z 476.9(M+H+) ES-MS: m/z 476.9 (M+H + )

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-11Example 7-11 5-氯-1'-(4-氯苄基)-5'-甲基-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮之外消旋(R,R)與(S,S)或外消旋(R,S)與(S,R) 5-Chloro-1'-(4-chlorobenzyl)-5'-methyl-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b] Pyridine]-2,6'(1'H)-dione racemic (R,R) and (S,S) or racemic (R,S) and (S,R)

使用通用程序以及上述實例之程序來製備5-氯-1'-(4-氯苄基)-5'-甲基-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮之外消旋(R,R)與(S,S)或外消旋(R,S)與(S,R)，且特徵資料為： 1H NMR(400MHz,DMSO-d6)：δ ppm 0.75(d,J=6.6Hz,3H),3.0(d,J=6.6Hz,1H),5.28(q,J=16.2Hz,2H),6.85(s,1H),6.93(d,J=8.3Hz,1H),6.97(s,1H),7.22(d,J=7.9Hz,2H),7.28(d,J=8.4Hz,2H),7.41(d,J=7.9Hz,1H),10.89(s,1H),11.19(s,1H)。 Preparation of 5-chloro-1'-(4-chlorobenzyl)-5'-methyl-5',7'-dihydrospiro[porphyrin-3,4'- using the general procedure and the procedure of the above examples. Pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione racemic (R,R) with (S,S) or racemic (R,S) (S, R), and the characteristic data are: 1 H NMR (400 MHz, DMSO-d 6 ): δ ppm 0.75 (d, J = 6.6 Hz, 3H), 3.0 (d, J = 6.6 Hz, 1H), 5.28 (q, J = 16.2 Hz, 2H), 6.85 (s, 1H), 6.93 (d, J = 8.3 Hz, 1H), 6.97 (s, 1H), 7.22 (d, J = 7.9 Hz, 2H), 7.28 (d, J = 8.4 Hz, 2H), 7.41 (d, J = 7.9 Hz, 1H), 10.89 (s, 1H), 11.19 (s, 1H).

MS：m/z 427.0(M+H)+。 MS: m/z 427.0 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備個別(R,R)、(S,S)、(R,S)及(S,R)對映異構體。 Individual (R,R), (S,S), (R,S) and (S,R) enantiomers can be prepared from the racemate by a procedure similar to the above examples. .

實例7-12Example 7-12 (R)-5-氯-1',7'-雙(4-氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-chloro-1',7'-bis(4-chlorobenzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4- b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為5-氯-1',7'-雙(4-氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H NMR(400MHz,DMSO-d6)：δ ppm 2.77(d,J=15.4Hz,1H),3.47(d,J=15.4Hz,1H),4.92(d,J=17.6Hz,1H),5.13(d,J=17.6Hz,1H),5.19(s,2H),6.84(d,J=8.4Hz,2H),6.94(s,1H),6.96(s,1H),7.27-7.4(m,7H),10.75(s,1H)。 The above-mentioned title compound racemate, which is 5-chloro-1',7'-bis(4-chlorobenzyl)-5',7'-dihydrospiro, was prepared using the general procedure and the procedure of the above example. Porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristics are: 1 H NMR (400 MHz, DMSO- d6 ) : δ ppm 2.77 (d, J = 15.4 Hz, 1H), 3.47 (d, J = 15.4 Hz, 1H), 4.92 (d, J = 17.6 Hz, 1H), 5.13 (d, J = 17.6 Hz, 1H) , 5.19 (s, 2H), 6.84 (d, J = 8.4 Hz, 2H), 6.94 (s, 1H), 6.96 (s, 1H), 7.27-7.4 (m, 7H), 10.75 (s, 1H).

MS：m/z 537.1(M+H)+。 MS: m/z 537.1 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-13Example 7-13 5-氯-1'-(1-(4-氯苯基)乙基)-7'-甲基-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮之外消旋(R,R)與(S,S)或外消旋(R,S)與(S,R) 5-Chloro-1'-(1-(4-chlorophenyl)ethyl)-7'-methyl-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[ 3,4-b]pyridine]-2,6'(1'H)-dione racemic (R,R) with (S,S) or racemic (R,S) and (S,R )

使用通用程序以及上述實例之程序來製備5-氯-1'-(1-(4-氯苯基)乙基)-7'-甲基-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮之外消旋(R,R)與(S,S)或外消旋(R,S)與(S,R)，且特徵資料為：H-NMR(400MHz,DMSO-d6)：δ ppm 1.81(d,J=6.6Hz,3H),2.61(d,J=14.9Hz,1H),3.14(d,J=15.3Hz,1H),3.36(s,3H),5.90(m,1H),6.91(d,J=8.4Hz,1H),6.93(s,1H),7.19(d,J=8.3Hz,2H),7.27(s,1H),7.32(d,J=9.2Hz,1H),7.41(d,J=8.4Hz,2H),10.7(s,1H)。 Preparation of 5-chloro-1'-(1-(4-chlorophenyl)ethyl)-7'-methyl-5',7'-dihydrospiro[porphyrin using a general procedure and the procedure of the above examples -3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione racemic (R,R) and (S,S) or racemic (R, S) and (S, R), and the characteristic data are: H-NMR (400MHz, DMSO-d 6 ): δ ppm 1.81 (d, J = 6.6Hz, 3H), 2.61 (d, J = 14.9 Hz, 1H), 3.14 (d, J = 15.3 Hz, 1H), 3.36 (s, 3H), 5.90 (m, 1H), 6.91 (d, J = 8.4 Hz, 1H), 6.93 (s, 1H), 7.19 (d, J = 8.3 Hz, 2H), 7.27 (s, 1H), 7.32 (d, J = 9.2 Hz, 1H), 7.41 (d, J = 8.4 Hz, 2H), 10.7 (s, 1H).

MS：m/z 441.0(M+H)+。 MS: m/z 441.0 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備外消旋混合物之個別(R,R)、(S,S)、(R,S)及(S,R)對映異構體。 The individual (R, R), (S, S), (R, S) and (S, R) of the racemic mixture can be prepared from the racemate by the procedure similar to the above examples. Enantiomer.

實例7-14Example 7-14 (R)-5-溴-1'-(4-(三氟甲基)苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-bromo-1'-(4-(trifluoromethyl)benzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4 -b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為5-溴-1'-(4-(三氟甲基)苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H-NMR(400MHz,DMSO-d6)：δ ppm 2.54(d,1H，與DMSO合併)，3.14(d,J=16.3Hz,1H),5.39(ABq,J=15.9Hz,2H),6.82(s,1H),6.87(d,J=8.4Hz,1H),7.50-7.40(m,4H),7.74(d,J=8.4Hz,2H),10.62(s,1H),11.10(br s,1H)。 The above-mentioned title compound racemate, which is 5-bromo-1 '-(4-(trifluoromethyl)benzyl)-5',7'-dihydrospiro, is prepared using the general procedure and the procedure of the above example. [Porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristics are: 1 H-NMR (400 MHz, DMSO- d 6 ): δ ppm 2.54 (d, 1H, combined with DMSO), 3.14 (d, J = 16.3 Hz, 1H), 5.39 (ABq, J = 15.9 Hz, 2H), 6.82 (s, 1H), 6.87 ( d, J = 8.4 Hz, 1H), 7.50-7.40 (m, 4H), 7.74 (d, J = 8.4 Hz, 2H), 10.62 (s, 1H), 11.10 (br s, 1H).

MS m/z：491.0(M+H)+。 MS m/z: 491.0 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-15Example 7-15 (R)-5-氯-1'-((5-氯吡啶-2-基)甲基)-7'-甲基-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-chloro-1'-((5-chloropyridin-2-yl)methyl)-7'-methyl-5',7'-dihydrospiro[porphyrin-3,4' -pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外 消旋物，其為5-氯-1'-((5-氯吡啶-2-基)甲基)-7'-甲基-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H NMR(400MHz,CD3OD)：δ ppm 2.74(d,J=15.4Hz,1H)。3.20(d,J=15.8Hz,1H),3.45(s,3H),5.63(s,2H),6.96(s,1H),6.97(d,J=5.7Hz,1H),7.19(d,J=8.8Hz,1H),7.28(s,1H),7.31(d,J=8.3Hz,1H),7.85(d,J=6.6Hz,1H),8.55(s,1H)。 The above-mentioned title compound racemate, which is 5-chloro-1 '-((5-chloropyridin-2-yl)methyl)-7'-methyl-5, was prepared using the general procedure and procedure of the above example. ',7'-Dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristic data are: 1 H NMR (400 MHz, CD 3 OD): δ δ 2.74 (d, J = 15.4 Hz, 1H). 3.20 (d, J = 15.8 Hz, 1H), 3.45 (s, 3H), 5.63 (s, 2H), 6.96 (s, 1H), 6.97 (d, J = 5.7 Hz, 1H), 7.19 (d, J = 8.8 Hz, 1H), 7.28 (s, 1H), 7.31 (d, J = 8.3 Hz, 1H), 7.85 (d, J = 6.6 Hz, 1H), 8.55 (s, 1H).

MS：m/z 428.0(M+H)+。 MS: m/z 428.0 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-16Example 7-16 (R)-5-氯-1'-((5-(三氟甲基)吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-chloro-1'-((5-(trifluoromethyl)pyridin-2-yl)methyl)-5',7'-dihydrospiro[porphyrin-3,4'- Pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為5-氯-1'-((5-(三氟甲基)吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H-NMR(400MHz,DMSO-d6)：δ ppm 2.51(d，與DMSO合併)，1H),3.13(d,J=16.0Hz,1H),5.52(ABq,J=16.8、2H),6.85(s,1H),6.92(d,J=7.6Hz,1H),7.19-7.34(m,3H),8.23(d,J=6.4Hz,1H),8.971(s,1H),10.638(s,1H),11.06(br s,1H)。 The above-mentioned title compound racemate, which is 5-chloro-1 '-((5-(trifluoromethyl)pyridin-2-yl)methyl)-5', was prepared using the general procedure and procedure of the above example. , 7'-Dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristic data is: 1 H -NMR (400MHz, DMSO-d 6 ): δ ppm 2.51 (d, merged with DMSO), 1H), 3.13 (d , J = 16.0Hz, 1H), 5.52 (ABq, J = 16.8,2H), 6.85 ( s, 1H), 6.92 (d, J = 7.6 Hz, 1H), 7.19-7.34 (m, 3H), 8.23 (d, J = 6.4 Hz, 1H), 8.971 (s, 1H), 10.638 (s, 1H) ), 11.06 (br s, 1H).

MS：m/z 448.0(M+H)+。 MS: m / z 448.0 (M + H) +.

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-17Example 7-17 (R)-5-氯-1'-((5-甲氧基吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-chloro-1'-((5-methoxypyridin-2-yl)methyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazole [3,4-b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為5-氯-1'-((5-甲氧基吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H NMR(400MHz,DMSO-d6)：δ ppm 3.12(d,J=15.8Hz,1H),3.56(br s,1H),3.81(s,3H),5.32(ABq,J=15.9Hz,2H),6.77(s,1H),6.91(d,J=8.4Hz,1H),7.03(d,J=8.8Hz,1H),7.29-7.42(m,3H),8.26(d,J=2.7Hz,1H),10.62(s,1H),11.03(br s,1H)。 The above-mentioned title compound racemate, which is 5-chloro-1 '-((5-methoxypyridin-2-yl)methyl)-5', 7', was prepared using the general procedure and procedure of the above example. - Dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristics are: 1 H NMR (400 MHz , DMSO- d6 ): δ ppm 3.12 (d, J = 15.8 Hz, 1H), 3.56 (br s, 1H), 3.81 (s, 3H), 5.32 (ABq, J = 15.9 Hz, 2H), 6.77 (s , 1H), 6.91 (d, J = 8.4 Hz, 1H), 7.03 (d, J = 8.8 Hz, 1H), 7.29-7.42 (m, 3H), 8.26 (d, J = 2.7 Hz, 1H), 10.62 (s, 1H), 11.03 (br s, 1H).

MS：m/z 410.1(M+H)+。 MS: m/z 410.1 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-18Example 7-18 (R)-5-(三氟甲氧基)-1'-((5-(三氟甲基)吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-(trifluoromethoxy)-1'-((5-(trifluoromethyl)pyridin-2-yl)methyl)-5',7'-dihydrospiro[porphyrin -3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為5-(三氟甲氧基)-1'-((5-(三氟甲基)吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H-NMR(400MHz,DMSO-d6)：δ ppm 2.67(d,J=20.6Hz,1H),3.30(d,J=15.8Hz,1H),5.52(d,J=6.6Hz,2H),6.81(s,1H),6.9(s,1H),7.00(s,1H),7.27(d,J=8.3Hz,1H),7.31(s,2H),8.21(dd,J=2.2Hz and 2.2Hz,1H),8.96(s,1H),10.60(s,1H),11.07(br s,1H)。 The above-mentioned title compound racemate, which is 5-(trifluoromethoxy)-1'-((5-(trifluoromethyl)pyridin-2-yl), was prepared using the general procedure and procedure of the above example. Methyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and The characterization data are: 1 H-NMR (400 MHz, DMSO-d 6 ): δ ppm 2.67 (d, J = 20.6 Hz, 1H), 3.30 (d, J = 15.8 Hz, 1H), 5.52 (d, J = 6.6 Hz, 2H), 6.81 (s, 1H), 6.9 (s, 1H), 7.00 (s, 1H), 7.27 (d, J = 8.3 Hz, 1H), 7.31 (s, 2H), 8.21 (dd, J =2.2 Hz and 2.2 Hz, 1H), 8.96 (s, 1H), 10.60 (s, 1H), 11.07 (br s, 1H).

MS：m/z 498.0(M+H)+。 MS: m/z 498.0 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-19Example 7-19 (R)-5-(三氟甲基)-1'-((5-(三氟甲基)吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-(trifluoromethyl)-1'-((5-(trifluoromethyl)pyridin-2-yl)methyl)-5',7'-dihydrospiro[porphyrin- 3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為5-(三氟甲基)-1'-((5-(三氟甲基)吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H-NMR(400MHz,DMSO-d6)：δ ppm 2.61(d,J=16.4Hz,1H),3.22(d,J=16.4Hz,1H),5.53(ABq,J=16.4Hz,2H),6.83(s,1H),7.09(d,J=7.6Hz,1H),7.23(d,J=8.4Hz,1H),7.641(m,2H),8.23(d,J=7.6Hz,1H),8.96(s,1H),10.91(s,1H),11.09(br s,1H)。 The above-mentioned title compound racemate, which is 5-(trifluoromethyl)-1'-((5-(trifluoromethyl)pyridin-2-yl), was prepared using the general procedure and procedure of the above example. -5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and characterized The data are: 1 H-NMR (400 MHz, DMSO-d 6 ): δ ppm 2.61 (d, J = 16.4 Hz, 1H), 3.22 (d, J = 16.4 Hz, 1H), 5.53 (ABq, J = 16.4 Hz , 2H), 6.83 (s, 1H), 7.09 (d, J = 7.6 Hz, 1H), 7.23 (d, J = 8.4 Hz, 1H), 7.641 (m, 2H), 8.23 (d, J = 7.6 Hz) , 1H), 8.96 (s, 1H), 10.91 (s, 1H), 11.09 (br s, 1H).

MS：m/z 480.1(M-H)-。 MS: m/z 480.1 (MH) - .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-20Example 7-20 (R)-1'-((5-氯吡啶-2-基)甲基)-5-乙基-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-1'-((5-chloropyridin-2-yl)methyl)-5-ethyl-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[ 3,4-b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為1'-((5-氯吡啶-2-基)甲基)-5-乙基-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H NMR(400MHz,DMSO-d6)：δ ppm1.14(t,J=7.6、3H),2.5(m,3H、與DMSO合併)，2.98(d,J=15.6Hz,1H),5.40(ABq,J=16.4Hz,2H),6.7(s,1H),6.819(d,J=7.5Hz,1H),7.07(m,3H),7.94(t,1H,J=1.6Hz),8.62(s,1H),10.42(s,1H),11.03(br s,1H)。 The above-mentioned title compound racemate, which is 1'-((5-chloropyridin-2-yl)methyl)-5-ethyl-5',7'-, was prepared using the general procedure and the procedure of the above example. Dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristics are: 1 H NMR (400 MHz, DMSO-d 6 ): δ ppm 1.14 (t, J = 7.6, 3H), 2.5 (m, 3H, combined with DMSO), 2.98 (d, J = 15.6 Hz, 1H), 5.40 (ABq, J = 16.4) Hz, 2H), 6.7 (s, 1H), 6.819 (d, J = 7.5 Hz, 1H), 7.07 (m, 3H), 7.94 (t, 1H, J = 1.6 Hz), 8.62 (s, 1H), 10.42 (s, 1H), 11.03 (br s, 1H).

MS：m/z 408.0(M+H)+。 MS: m/z 408.0 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-21Example 7-21 (R)-5-溴-1'-((5-氯吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-bromo-1'-((5-chloropyridin-2-yl)methyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3 ,4-b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為5-溴-1'-((5-氯吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H-NMR(400MHz,DMSO-d6)：δ ppm 2.56(d,1H，與DMSO合併)，3.12(d,J=6.4Hz,1H),5.42(ABq,J=16.4Hz,2H),6.82(s,1H),6.87(d,J=8.8Hz,1H),7.06(d,J=8.4,1H),7.43(d,J=6.8Hz),7.93(dd,J=2.4,2.0Hz,1H),8.61(s,1H),10.63(s,1H),11.03(br s,1H)。 The above-mentioned title compound racemate, which is 5-bromo-1 '-((5-chloropyridin-2-yl)methyl)-5', 7'-di, was prepared using the general procedure and procedure of the above example. Hydrogen spiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristics are: 1 H-NMR (400 MHz, DMSO-d 6 ): δ ppm 2.56 (d, 1H, combined with DMSO), 3.12 (d, J = 6.4 Hz, 1H), 5.42 (ABq, J = 16.4 Hz, 2H), 6.82 (s, 1H), 6.87 (d, J = 8.8 Hz, 1H), 7.06 (d, J = 8.4, 1H), 7.43 (d, J = 6.8 Hz), 7.93 (dd, J = 2.4, 2.0 Hz, 1H), 8.61 (s) , 1H), 10.63 (s, 1H), 11.03 (br s, 1H).

MS：m/z 458.0(M+H)+。 MS: m/z 458.0 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-22Example 7-22 (R)-1'-((5-氯吡啶-2-基)甲基)-5-(三氟甲氧基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-1'-((5-chloropyridin-2-yl)methyl)-5-(trifluoromethoxy)-5',7'-dihydrospiro[porphyrin-3,4' -pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為1'-((5-氯吡啶-2-基)甲基)-5-(三氟甲氧基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H-NMR(400MHz,DMSO-d6)：δ 2.50(d,J=1.6Hz,1H),3.157(d,J=6.2Hz,1H),5.40(ABq,J=16.3Hz,2H),6.78(s,1H),6.98(d J=8.4Hz,1H)7.07(d J=8.40Hz 1H),7.28(m,2H),7.93(dd J=10.8Hz,1H),8.615(d,J=2.4Hz,1H),10.32(s,1H),11.09(br s,1H)。 The above-mentioned title compound racemate, which is 1'-((5-chloropyridin-2-yl)methyl)-5-(trifluoromethoxy)-5, was prepared using the general procedure and procedure ',7'-Dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristic data are: 1 H-NMR (400MHz, DMSO- d 6): δ 2.50 (d, J = 1.6Hz, 1H), 3.157 (d, J = 6.2Hz, 1H), 5.40 (ABq, J = 16.3Hz, 2H), 6.78 (s, 1H), 6.98 (d J = 8.4 Hz, 1H) 7.07 (d J = 8.40 Hz 1H), 7.28 (m, 2H), 7.93 (dd J = 10.8 Hz, 1H), 8.615 (d, J = 2.4 Hz, 1H), 10.32 (s, 1H), 11.09 (br s, 1H).

MS：m/z 464.2(M+H)+。 MS: m/z 464.2 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-23Example 7-23 (R)-6-氯-1'-((5-氯吡啶-2-基)甲基)-5-氟-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-6-chloro-1'-((5-chloropyridin-2-yl)methyl)-5-fluoro-5',7'-dihydrospiro[porphyrin-3,4'-pyridyl Oxazo[3,4-b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為6-氯-1'-((5-氯吡啶-2-基)甲基)-5-氟-5',7'-二氫螺[吲哚啉- 3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H-NMR(400MHz,DMSO-d6)：δ ppm 2.54(d,1H，與DMSO合併)，3.16(d,J=16.2Hz,1H),5.40(d,J=11.0Hz,2H),6.85(s,1H),7.01-7.17(m,2H),7.43(d,J=8.8Hz,1H),7.93(dd,J=2.6,2.7Hz,1H),8.61(d,J=2.2Hz,1H),10.61(br s,1H),11.04(br s,1H)。 The above-mentioned title compound racemate, which is 6-chloro-1 '-((5-chloropyridin-2-yl)methyl)-5-fluoro-5', was prepared using the general procedure and the procedure of the above example. 7'-Dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristic data is: 1 H- NMR (400 MHz, DMSO-d 6 ): δ ppm 2.54 (d, 1H, combined with DMSO), 3.16 (d, J = 16.2 Hz, 1H), 5.40 (d, J = 11.0 Hz, 2H), 6.85 (s) , 1H), 7.01-7.17 (m, 2H), 7.43 (d, J = 8.8 Hz, 1H), 7.93 (dd, J = 2.6, 2.7 Hz, 1H), 8.61 (d, J = 2.2 Hz, 1H) , 10.61 (br s, 1H), 11.04 (br s, 1H).

MS：m/z 432.0(M+H)+。 MS: m/z 432.0 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-24Example 7-24 (R)-5-氯-1'-((3,5-二氯吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-chloro-1'-((3,5-dichloropyridin-2-yl)methyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazole And [3,4-b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為5-氯-1'-((3,5-二氯吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H-NMR(400MHz,DMSO-d6)：δ ppm 2.55(d,1H，與DMSO合併)，3.01(d,J=15.6Hz,1H),5.46(s,2H),6.74(s,1H),6.89(d,J=8.4Hz,1H),7.23-7.26(m,2H),8.26(s,1H),8.53(s,1H),10.6(s,1H),10.99(br s,1H)。 The above-mentioned title compound racemate, which is 5-chloro-1 '-((3,5-dichloropyridin-2-yl)methyl)-5',7, was prepared using the general procedure and procedure of the above example. '-Dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristics are: 1 H-NMR (400MHz, DMSO-d 6 ): δ ppm 2.55 (d, 1H, combined with DMSO), 3.01 (d, J = 15.6 Hz, 1H), 5.46 (s, 2H), 6.74 (s, 1H), 6.89 ( d, J = 8.4 Hz, 1H), 7.23 - 7.26 (m, 2H), 8.26 (s, 1H), 8.53 (s, 1H), 10.6 (s, 1H), 10.99 (br s, 1H).

MS m/z：446.2(M+H)+。 MS m/z: 446.2 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來 製備標題化合物。 A program similar to the above example can be used to analyze the palmarity by the racemate. The title compound was prepared.

實例7-25Example 7-25 (R)-5-氯-1'-(4-氯苄基)-1-甲基-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮 (R)-5-chloro-1'-(4-chlorobenzyl)-1-methyl-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4 -b]pyridine]-2,6'(1'H)-dione

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為5-氯-1'-(4-氯苄基)-1-甲基-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮，且特徵資料為：1H NMR(400MHz,DMSO-d6)：δ ppm 2.54(d，峰與DMSO合併，1H),3.10-3.20(m,4H),5.28(ABq,J=15.9Hz,2H),6.77(s,1H),7.1(d,J=8.3Hz,1H),7.25(d,J=7.9Hz,2H),7.35-7.45(m,4H),11.11(br s,1H)。 The above-mentioned title compound racemate, which is 5-chloro-1 '-(4-chlorobenzyl)-1-methyl-5',7'-dihydrospiro, is prepared using the general procedure and the procedure of the above example. [Porphyrin-3,4'-pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione, and the characteristics are: 1 H NMR (400 MHz, DMSO- d6) ): δ ppm 2.54 (d, peak combined with DMSO, 1H), 3.10-3.20 (m, 4H), 5.28 (ABq, J = 15.9 Hz, 2H), 6.77 (s, 1H), 7.1 (d, J = 8.3 Hz, 1H), 7.25 (d, J = 7.9 Hz, 2H), 7.35-7.45 (m, 4H), 11.11 (br s, 1H).

MS：m/z 427.0(M+H)+。 MS: m/z 427.0 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例7-26Example 7-26 (R)-5-氯-1'-(4-氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-6'(1'H)-酮 (R)-5-chloro-1'-(4-chlorobenzyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b]pyridine] -6'(1'H)-ketone

使用通用程序以及上述實例之程序來製備以上標題化合物之外消旋物，其為5-氯-1'-(4-氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-6'(1'H)-酮，且特徵資料為：1H NMR(400MHz,CD3OD)：δ 2.78(AB q,J=16.4Hz,2H),3.50(AB q,J=9.2Hz,2H),5.28(s,2H),6.65(d,J=8.0Hz,1H),6.90(d,J=2.0Hz,1H),7.02(dd,J=2.0Hz,1H),7.15(d,J=8.4Hz,2H),7.23(s,1H),7.35(d,J=8.4Hz,2H)。 The above-mentioned title compound racemate, which is 5-chloro-1 '-(4-chlorobenzyl)-5',7'-dihydrospiro[porphyrin-, was prepared using the general procedure and procedure of the above example. 3,4'-pyrazolo[3,4-b]pyridine]-6'(1'H)-one, and the characteristics are: 1 H NMR (400 MHz, CD 3 OD): δ 2.78 (AB q, J = 16.4 Hz, 2H), 3.50 (AB q, J = 9.2 Hz, 2H), 5.28 (s, 2H), 6.65 (d, J = 8.0 Hz, 1H), 6.90 (d, J = 2.0 Hz, 1H) ), 7.02 (dd, J = 2.0 Hz, 1H), 7.15 (d, J = 8.4 Hz, 2H), 7.23 (s, 1H), 7.35 (d, J = 8.4 Hz, 2H).

MS：m/z 399.0(M+H)+。 MS: m/z 399.0 (M + H) + .

可使用與上述實例類似之程序，經由對掌性解析由外消旋物來製備標題化合物。 The title compound can be prepared from the racemate by parsing the palms using procedures similar to those described above.

實例8-1及實例8-2Example 8-1 and Example 8-2 製備5'-氯-3-(1-(4-(三氟甲基)苯基)乙基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮之(R,R)與(S,S)外消旋混合物或(R,S)與(S,R)外消旋混合物(實例8-1)；及製備5'-氯-3-(1-(4-(三氟甲基)苯基)乙基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮之(R,S)與(S,R)外消旋混合物或(R,R)與(S,S)外消旋混合物(實例8-2) Preparation of 5'-chloro-3-(1-(4-(trifluoromethyl)phenyl)ethyl)-4,6-dihydrospiro[[1,2,3]triazolo[4,5- b] pyridine-7,3'-carboline]-2',5(3H)-dione (R,R) and (S,S) racemic mixture or (R,S) and (S, R) racemic mixture (Example 8-1); and preparation of 5'-chloro-3-(1-(4-(trifluoromethyl)phenyl)ethyl)-4,6-dihydrospiro[[ 1,2,3]triazolo[4,5-b]pyridine-7,3'-carboline]-2',5(3H)-dione (R,S) and (S,R) Racemic mixture or (R, R) and (S, S) racemic mixture (Example 8-2)

步驟1：製備1-(4-(三氟甲基)苯基)乙醇 Step 1: Preparation of 1-(4-(trifluoromethyl)phenyl)ethanol

在0℃下，向4'-(三氟甲基)苯乙酮(10g，53.15mmol)於乙醇(55mL)中之冷卻溶液中以小份添加NaBH4且將所得混合物在室溫下攪拌4h。以1N HCl(50mL)中止反應混合物且以CH2Cl2(3×100mL)萃取。將經合併之有機層以飽和NaHCO3、鹽水洗滌，經無水Na2SO4乾燥且在減壓下濃縮以提供9.5g(94%)呈淺黃色液體狀之1-(4-(三氟甲基)苯基)乙醇。 Add NaBH 4 in small portions to a cooled solution of 4'-(trifluoromethyl)acetophenone (10 g, 53.15 mmol) in ethanol (55 mL). . In 1N HCl (50mL) and the reaction mixture was quenched in CH 2 Cl 2 (3 × 100mL ) and extracted. The combined organic layers were washed with saturated NaHCO 3, brine, dried over anhydrous Na 2 SO 4 dried and concentrated to afford 9.5g (94%) as a pale yellow liquid of 1- (4- (trifluoromethyl under reduced pressure Base) phenyl)ethanol .

1H-NMR(400MHz,CDCl3)；δ 7.6(d,J=8.0Hz,2H),7.49(d,J=8.4Hz,2H),4.94-5.0(m,1H),1.86(s,1H),1.51(d,J=6.8Hz,3H)。 1 H-NMR (400 MHz, CDCl 3 ); δ 7.6 (d, J = 8.0 Hz, 2H), 7.49 (d, J = 8.4 Hz, 2H), 4.94-5.0 (m, 1H), 1.86 (s, 1H) ), 1.51 (d, J = 6.8 Hz, 3H).

步驟2：製備1-(1-溴乙基)-4-(三氟甲基)苯 Step 2: Preparation of 1-(1-bromoethyl)-4-(trifluoromethyl)benzene

在0℃下，向1-(4-(三氟甲基)苯基)乙醇(2g，10.52mmol)於CH2Cl2(30mL)中之冷卻溶液中緩慢添加三溴化磷(1.1mL，11.58mmol)且使所得混合物在室溫下攪拌2h。將反應團以CH2Cl2(100mL) 稀釋且以水、飽和NaHCO3、水及鹽水洗滌。經無水Na2SO4乾燥所得有機層且在減壓下濃縮以提供2.6g(98%)呈淺黃色液體狀之1-(1-溴乙基)-4-(三氟甲基)苯。 At 0 ℃, phosphorus tribromide was slowly added a solution of 1- (4- (trifluoromethyl) phenyl) ethanol (2g, 10.52mmol) in CH 2 Cl 2 (30mL) in a cool solution (1.1mL, 11.58 mmol) and the resulting mixture was stirred at room temperature for 2 h. The reaction was diluted with radical CH 2 Cl 2 (100mL) and water, washed with saturated NaHCO 3, water and brine. The resulting organic layers were dried over anhydrous Na 2 SO and concentrated to provide 2.6g (98%) as a pale yellow liquid of 1- (1-bromoethyl) -4- (trifluoromethyl) benzene under reduced pressure.

1H-NMR(400MHz,CDCl3)：δ 7.6(d,J=8.4Hz,2H),7.55(d,J=8.4Hz,2H),5.19(q,J=6.9Hz,1H),2.05(d,J=6.8Hz,3H)。 1 H-NMR (400MHz, CDCl 3): δ 7.6 (d, J = 8.4Hz, 2H), 7.55 (d, J = 8.4Hz, 2H), 5.19 (q, J = 6.9Hz, 1H), 2.05 ( d, J = 6.8 Hz, 3H).

步驟3：製備1-(1-疊氮基乙基)-4-(三氟甲基)苯 Step 3: Preparation of 1-(1-azidoethyl)-4-(trifluoromethyl)benzene

向1-(1-溴乙基)-4-(三氟甲基)苯(13g，51.37mmol)於DMSO(130mL)中之溶液中添加疊氮化鈉(3.34g，128.46mmol)且將所得混合物在室溫下攪拌5h。以水中止反應團且以乙酸乙酯(3×200mL)萃取。將經合併之有機層以水、鹽水洗滌，經無水Na2SO4乾燥且在減壓下濃縮以提供10g(90%)呈無色液體狀之1-(1-疊氮基乙基)-4-(三氟甲基)苯。 Add sodium azide (3.34 g, 128.46 mmol) to a solution of 1-(1-bromoethyl)-4-(trifluoromethyl)benzene (13 g, 51.37 mmol) in EtOAc (EtOAc) The mixture was stirred at room temperature for 5 h. The reaction mass was quenched with water and extracted with ethyl acetate (3×200 mL). The combined organic layers were washed with water, brine, dried over anhydrous Na 2 SO 4 dried and concentrated to afford 10g (90%) as a colorless liquid of 1- (1-azido-ethyl) -4 under reduced pressure -(Trifluoromethyl)benzene .

1H-NMR(400MHz,CDCl3)：δ 7.64(d,J=8.0Hz,2H),7.44(d,J=8.4Hz,2H),4.68(q,J=6.8Hz,1H),1.55(d,J=6.4Hz,3H)。 1 H-NMR (400 MHz, CDCl 3 ): δ 7.64 (d, J = 8.0 Hz, 2H), 7.44 (d, J = 8.4 Hz, 2H), 4.68 (q, J = 6.8 Hz, 1H), 1.55 ( d, J = 6.4 Hz, 3H).

步驟4：製備5-胺基-1-(1-(4-(三氟甲基)苯基)乙基)-1H-1,2,3-***-4-甲酸 Step 4: Preparation of 5-amino-1-(1-(4-(trifluoromethyl)phenyl)ethyl)-1H-1,2,3-triazole-4-carboxylic acid

向冷卻至0℃之Na(0.214g，9.29mmol)於乙醇(15mL)中之透明溶液中添加氰基乙酸(0.395g，4.65mmol)、1-(1-疊氮基乙基)-4-(三氟 甲基)苯(1g，4.65mmol)且將所得混合物在回流溫度下攪拌24h。以冰中止反應混合物且在減壓下餾出溶劑。將殘餘物溶解於水中且以濃HCl酸化溶液。藉由過濾收集所得固體、以***洗滌且在減壓下乾燥以提供200mg(12%基於LCMS至85%)呈奶白色固體狀之5-胺基-1-(1-(4-(三氟甲基)苯基)乙基)-1H-1,2,3-***-4-甲酸。 Add cyanoacetic acid (0.395 g, 4.65 mmol), 1-(1-azidoethyl)-4- to a clear solution of Na (0.214 g, 9.29 mmol) in EtOAc (15 mL). (Trifluoromethyl )benzene (1 g, 4.65 mmol) and the mixture was stirred at reflux temperature for 24 h. The reaction mixture was quenched with ice and the solvent was evaporated under reduced pressure. The residue was dissolved in water and the solution was acidified with cone. HCl. The resulting solid was collected by filtration, washed with ether and dried under reduced pressure, and to provide 200mg (12% to 85% based on the LCMS) as a white solid milk of 5-amino-1- (1- (4- (trifluoromethyl Methyl)phenyl)ethyl)-1H-1,2,3-triazole-4-carboxylic acid .

1H-NMR(400MHz,DMSO-d 6 )：δ 12.25(br s,1H),7.73(d,J=8.4Hz,2H),7.45(d,J=8.0Hz,2H),6.46(s,2H),5.81(q,J=7.2Hz,1H),1.88(d,J=6.8Hz,3H)；LCMS(ESI)：m/z 301.0[M+H]+。 1 H-NMR (400MHz, DMSO- d 6): δ 12.25 (br s, 1H), 7.73 (d, J = 8.4Hz, 2H), 7.45 (d, J = 8.0Hz, 2H), 6.46 (s, 2H), 5.81 (q, J = 7.2 Hz, 1H), 1.88 (d, J = 6.8 Hz, 3H); LCMS (ESI): m/z 301.0 [M+H] + .

步驟5：製備1-(1-(4-(三氟甲基)苯基)乙基)-1H-1,2,3-***-5-胺 Step 5: Preparation of 1-(1-(4-(trifluoromethyl)phenyl)ethyl)-1H-1,2,3-triazole-5-amine

將5-胺基-1-(1-(4-(三氟甲基)苯基)乙基)-1H-1,2,3-***-4-甲酸(1g，3.33mmol)與N,N-二甲基苯胺(7mL)之混合物在200℃之預熱油浴中攪拌15min。直接藉由使用石油醚中35-50% EtOAc之溶劑梯度作為溶離劑，經矽膠(100-200目)管柱層析純化反應團以提供200mg(23%)呈奶白色固體狀之1-(1-(4-(三氟甲基)苯基)乙基)-1H-1,2,3-***-5-胺。 5-Amino-1-(1-(4-(trifluoromethyl)phenyl)ethyl)-1H-1,2,3-triazole-4-carboxylic acid (1 g, 3.33 mmol) with N , A mixture of N -dimethylaniline (7 mL) was stirred in a preheated oil bath at 200 °C for 15 min. Directly by the use of a solvent gradient of 35-50% EtOAc in petroleum ether as the eluent, dried over silica gel (100-200 mesh) and purified by column chromatography to provide a radical reaction 200mg (23%) of a white solid milk 1- ( 1-(4-(Trifluoromethyl)phenyl)ethyl)-1H-1,2,3-triazole-5-amine .

1H-NMR(400MHz,DMSO-d 6 )：δ 7.71(d,J=8.0Hz,2H),7.41(d,J=8.0Hz,2H),6.83(s,1H),5.71(q,J=6.9Hz,1H),5.51(s,2H),1.85(d,J=7.2Hz,3H)；LCMS(ESI)：m/z 257.0[M+H]+。 1 H-NMR (400 MHz, DMSO- d 6 ): δ 7.71 (d, J = 8.0 Hz, 2H), 7.41 (d, J = 8.0 Hz, 2H), 6.83 (s, 1H), 5.71 (q, J = 6.9 Hz, 1H), 5.51 (s, 2H), 1.85 (d, J = 7.2 Hz, 3H); LCMS (ESI): m/z 257.0 [M+H] + .

步驟6：製備5'-氯-3-(1-(4-(三氟甲基)苯基)乙基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮 Step 6: Preparation of 5'-chloro-3-(1-(4-(trifluoromethyl)phenyl)ethyl)-4,6-dihydrospiro[[1,2,3]triazolo[4 ,5-b]pyridine-7,3'-carboline]-2',5(3H)-dione

將1-(1-(4-(三氟甲基)苯基)乙基)-1H-1,2,3-***-5-胺(200mg，0.78mmol)、5-氯靛紅(142mg，0.78mmol)及麥氏酸(112mg，0.78mmol)於乙酸(10mL)中之混合物在100℃之預熱油浴中攪拌4h。濃縮反應團且向殘餘物中添加水。藉由過濾收集所得固體，以水洗滌、繼而以己烷洗滌且在減壓下乾燥。對於D1而言使用石油醚中40-50% EtOAc之溶劑梯度作為溶離劑，且對於D2而言使用石油醚中60-80% EtOAc之溶劑梯度作為溶離劑，藉由經矽膠(100-200目)管柱層析分離非對映異構體。此外，以CH2Cl2/戊烷洗滌D1且以甲醇洗滌D2以提供30mg呈紅色固體狀之5'-氯-3-(1-(4-(三氟甲基)苯基)乙基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮(實例8-1)及35mg呈奶白色固體狀之5'-氯-3-(1-(4-(三氟甲基)苯基)乙基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮(實例8-2)。 1-(1-(4-(Trifluoromethyl)phenyl)ethyl)-1H-1,2,3-triazole-5-amine (200 mg, 0.78 mmol), 5-chloroindole (142 mg) , 0.78 mmol) and a mixture of the crude acid (112 mg, 0.78 mmol) in acetic acid (10 mL) was stirred in a preheated oil bath at 100 ° C for 4 h. The reaction mass was concentrated and water was added to the residue. The obtained solid was collected by filtration, washed with water, then washed with hexane and dried under reduced pressure. A solvent gradient of 40-50% EtOAc in petroleum ether was used as the eluent for D1, and a solvent gradient of 60-80% EtOAc in petroleum ether was used as the dissolving agent for D2 by gelatin (100-200 mesh) Column chromatography separates the diastereomers. Further, D1 was washed with CH 2 Cl 2 /pentane and D2 was washed with methanol to give 30 mg of 5'-chloro-3-(1-(4-(trifluoromethyl)phenyl)ethyl) as a red solid. -4,6-dihydrospiro[[1,2,3]triazolo[4,5-b]pyridine-7,3'-carboline]-2',5(3H)-dione (example) 8-1) and 35 mg of 5'-chloro-3-(1-(4-(trifluoromethyl)phenyl)ethyl)-4,6-dihydrospiro[[1,2] , 3] Triazolo[4,5-b]pyridine-7,3'-carboline]-2',5(3H)-dione (Example 8-2).

實例8-1之特徵資料展示如下：1H NMR(400MHz,DMSO-d6)：δ ppm 1.91(d,J=6.8Hz,3H),2.61(d,J=16.0Hz,1H),3.35(d,1H，與DMSO-水合併)，5.95(q,J=6.8Hz,1H),6.93(d,J=8.0Hz,1H),7.36-7.31(m,2H),7.54(d,J=8.4Hz,2H),7.78(d,J=8.4Hz,2H),10.70(s,1H),11.22(s,1H)。 The characteristics of Example 8-1 are shown below: 1 H NMR (400 MHz, DMSO-d 6 ): δ ppm 1.91 (d, J = 6.8 Hz, 3H), 2.61 (d, J = 16.0 Hz, 1H), 3.35 ( d, 1H, combined with DMSO-water), 5.95 (q, J = 6.8 Hz, 1H), 6.93 (d, J = 8.0 Hz, 1H), 7.36-7.31 (m, 2H), 7.54 (d, J = 8.4 Hz, 2H), 7.78 (d, J = 8.4 Hz, 2H), 10.70 (s, 1H), 11.22 (s, 1H).

MS：m/z 462.0(M+H)+。 MS: m/z 462.0 (M + H) + .

可藉由使用上述實例之程序分離外消旋混合物來製備實例8-1之個別(R,R)、(S,S)、(R,S)及(S,R)對映異構體。 The individual (R, R), (S, S), (R, S) and (S, R) enantiomers of Example 8-1 can be prepared by isolating the racemic mixture using the procedures of the above examples.

實例8-2之特徵資料展示如下：1H NMR(400MHz,DMSO-d6)：δ ppm 1.90(d,J=7.2Hz,3H),2.72(d,J=16.0Hz,1H),3.19(d,J=17.6Hz,1H),5.90(q,J=7.2Hz,1H),6.93(d,J=8.8Hz,1H),7.34-7.32(m,2H),7.56(d,J=8.4Hz,2H),7.77(d,J=8.0Hz,2H),10.71(s,1H),11.30(s,1H)。 The characteristics of Example 8-2 are shown below: 1 H NMR (400 MHz, DMSO-d 6 ): δ ppm 1.90 (d, J = 7.2 Hz, 3H), 2.72 (d, J = 16.0 Hz, 1H), 3.19 ( d, J = 17.6 Hz, 1H), 5.90 (q, J = 7.2 Hz, 1H), 6.93 (d, J = 8.8 Hz, 1H), 7.34 - 7.32 (m, 2H), 7.56 (d, J = 8.4 Hz, 2H), 7.77 (d, J = 8.0 Hz, 2H), 10.71 (s, 1H), 11.30 (s, 1H).

MS：m/z 462.0(M+H)+。 MS: m/z 462.0 (M + H) + .

可藉由使用上述實例之程序分離外消旋混合物來製備實例8-2之個別(R,S)、(S,R)、(R,R)及(S,S)對映異構體。 The individual (R, S), (S, R), (R, R) and (S, S) enantiomers of Example 8-2 can be prepared by isolating the racemic mixture using the procedures of the above examples.

使用通用程序以及上述實例之程序，由適當起始物質來製備以下化合物。 The following compounds were prepared from the appropriate starting materials using the general procedures and procedures of the above examples.

實例8-3Example 8-3 (5'-氯-3-(1-(4-氯苯基)丙基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮之(R,R)與(S,S)外消旋混合物或(R,S)與(S,R)外消旋混合物 (5'-Chloro-3-(1-(4-chlorophenyl)propyl)-4,6-dihydrospiro[[1,2,3]triazolo[4,5-b]pyridine-7 , 3'-porphyrin]-2',5(3H)-dione (R,R) and (S,S) racemic mixture or (R,S) and (S,R) racemic mixture

外消旋(R,R)與(S,S)或外消旋(R,S)與(S,R) Racemic (R, R) and (S, S) or racemic (R, S) and (S, R)

1H NMR(400MHz,DMSO-d6)：δ ppm 0.81(t,J=7.6Hz,3H),2.24-2.16(m,1H),2.43-2.32(m,1H),2.60(d,J=16.0Hz,1H),3.33(d,J=16.8Hz,1H),5.58(t,J=7.6Hz,1H),6.92(d,J=8.4Hz,1H),7.34-7.30(m,2H),7.49-7.43(m,4H),10.68(s,1H),11.22(s,1H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm 0.81 (t, J = 7.6Hz, 3H), 2.24-2.16 (m, 1H), 2.43-2.32 (m, 1H), 2.60 (d, J = 16.0 Hz, 1H), 3.33 (d, J = 16.8 Hz, 1H), 5.58 (t, J = 7.6 Hz, 1H), 6.92 (d, J = 8.4 Hz, 1H), 7.34-7.30 (m, 2H) , 7.49-7.43 (m, 4H), 10.68 (s, 1H), 11.22 (s, 1H).

MS：m/z 442.0(M+H)+。 MS: m/z 442.0 (M + H) + .

實例8-4Example 8-4 (5'-氯-3-(1-(4-氯苯基)丙基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮之(R,S)與(S,R)外消旋混合物或(R,R)與(S,S)外消旋混合物 (5'-Chloro-3-(1-(4-chlorophenyl)propyl)-4,6-dihydrospiro[[1,2,3]triazolo[4,5-b]pyridine-7 , 3'-porphyrin]-2',5(3H)-dione (R,S) and (S,R) racemic mixture or (R,R) and (S,S) racemic mixture

外消旋(R,S)與(S,R)或外消旋(R,R)與(S,S) Racemic (R, S) and (S, R) or racemic (R, R) and (S, S)

1H NMR(400MHz,DMSO-d6)：δ ppm 0.83(t,J=7.6Hz,3H),2.24-2.16(m,1H),2.40-2.32(m,1H),2.71(d,J=16.8Hz,1H),3.27(d,J=16.8Hz,1H),5.52(t,J=7.6Hz,1H),6.93(d,J=8.0Hz,1H),7.34-7.30(m,2H),7.49-7.43(m,4H),10.70(s,1H),11.30(s,1H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm 0.83 (t, J = 7.6Hz, 3H), 2.24-2.16 (m, 1H), 2.40-2.32 (m, 1H), 2.71 (d, J = 16.8 Hz, 1H), 3.27 (d, J = 16.8 Hz, 1H), 5.52 (t, J = 7.6 Hz, 1H), 6.93 (d, J = 8.0 Hz, 1H), 7.34-7.30 (m, 2H) , 7.49-7.43 (m, 4H), 10.70 (s, 1H), 11.30 (s, 1H).

MS：m/z 442.0(M+H)+。 MS: m/z 442.0 (M + H) + .

實例8-5Example 8-5 5-氯-1'-(1-(4-氯苯基)丙基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮之(R,R)與(S,S)外消旋混合物或(R,S)與(S,R)外消旋混合物 5-chloro-1'-(1-(4-chlorophenyl)propyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b] a racemic mixture of (R,R) with (S,S) or a racemic mixture of (R,S) and (S,R) of pyridine]-2,6'(1'H)-dione

外消旋(R,R)與(S,S)或外消旋(R,S)與(S,R) Racemic (R, R) and (S, S) or racemic (R, S) and (S, R)

1H NMR(400MHz,DMSO-d6)：δ ppm 0.78(t,J=7.6Hz,3H),2.05-2.00(m,1H),2.30-2.24(m,1H),2.44(d,J=15.6Hz,1H),3.12(d, J=16.0Hz,1H),5.47-5.41(m,1H),6.82(s,1H),6.91-6.89(m,1H),7.30-7.26(m,2H),7.44-7.38(m,4H),10.54(s,1H),10.97(s,1H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm 0.78 (t, J = 7.6Hz, 3H), 2.05-2.00 (m, 1H), 2.30-2.24 (m, 1H), 2.44 (d, J = 15.6 Hz, 1H), 3.12 (d, J = 16.0 Hz, 1H), 5.47-5.41 (m, 1H), 6.82 (s, 1H), 6.91-6.89 (m, 1H), 7.30-7.26 (m, 2H) ), 7.44 - 7.38 (m, 4H), 10.54 (s, 1H), 10.97 (s, 1H).

MS：m/z 441.3(M+H)+。 MS: m/z 441.3 (M + H) + .

實例8-6Example 8-6 5-氯-1'-(1-(4-氯苯基)丙基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮之(R,S)與(S,R)外消旋混合物或(R,R)與(S,S)外消旋混合物 5-chloro-1'-(1-(4-chlorophenyl)propyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4-b] a racemic mixture of (R,S) and (S,R) or a racemic mixture of (R,R) and (S,S) of pyridine]-2,6'(1'H)-dione

外消旋(R,S)與(S,R)或外消旋(R,R)與(S,S) Racemic (R, S) and (S, R) or racemic (R, R) and (S, S)

1H NMR(400MHz,DMSO-d6)：δ ppm 0.78(t,J=7.6Hz,3H),2.00-2.08(m,1H),2.26-2.19(m,1H),2.54(d,J=16.4Hz,1H),3.08(d,J=16.4Hz,1H),5.39-5.34(m,1H),6.84(s,1H),6.90(d,J=8.8Hz,1H),7.32-7.28(m,2H),7.40(d,J=8.4Hz,2H),7.47(d,J=8.4Hz,2H),10.57(s,1H),11.05(s,1H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm 0.78 (t, J = 7.6Hz, 3H), 2.00-2.08 (m, 1H), 2.26-2.19 (m, 1H), 2.54 (d, J = 16.4 Hz, 1H), 3.08 (d, J = 16.4 Hz, 1H), 5.39-5.34 (m, 1H), 6.84 (s, 1H), 6.90 (d, J = 8.8 Hz, 1H), 7.32 - 7.28 ( m, 2H), 7.40 (d, J = 8.4 Hz, 2H), 7.47 (d, J = 8.4 Hz, 2H), 10.57 (s, 1H), 11.05 (s, 1H).

MS：m/z 441.3(M+H)+。 MS: m/z 441.3 (M + H) + .

實例8-7Example 8-7 5-氯-1'-(1-(4-氯苯基)-2-甲基丙基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮之(R,R)與(S,S)外消旋混合物或(R,S)與(S,R)外消旋混合物 5-Chloro-1'-(1-(4-chlorophenyl)-2-methylpropyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3 , (b, S) racemic mixture of (B, pyridine)-2,6'(1'H)-dione, or (R,S) and (S,R) Spin mixture

外消旋(R,R)與(S,S)或外消旋(R,S)與(S,R) Racemic (R, R) and (S, S) or racemic (R, S) and (S, R)

1H NMR(400MHz,DMSO-d6)：δ ppm 0.74(t,J=6.8Hz,6H),2.43(J=16.0Hz,1H),2.61-2.55(m,1H),3.12(d,J=16.0Hz,1H),5.12(d,J=10.4Hz,1H),6.80(s,1H),6.90-6.88(m,1H),7.29-7.26(m,2H),7.43(d,J=8.4Hz,2H),7.55(d,J=8.4Hz,2H),10.52(s,1H),11.0(s,1H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm 0.74 (t, J = 6.8Hz, 6H), 2.43 (J = 16.0Hz, 1H), 2.61-2.55 (m, 1H), 3.12 (d, J =16.0 Hz, 1H), 5.12 (d, J = 10.4 Hz, 1H), 6.80 (s, 1H), 6.90-6.88 (m, 1H), 7.29-7.26 (m, 2H), 7.43 (d, J = 8.4 Hz, 2H), 7.55 (d, J = 8.4 Hz, 2H), 10.52 (s, 1H), 11.0 (s, 1H).

MS：m/z 454.9(M+H)+。 MS: m/z 454.9 (M + H) + .

實例8-8Example 8-8 5-氯-1'-(1-(4-氯苯基)-2-甲基丙基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮之(R,S)與(S,R)外消旋混合物或(R,R)與(S,S)外消旋混合物 5-Chloro-1'-(1-(4-chlorophenyl)-2-methylpropyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3 , (b, S) and (S, R) racemic mixtures or (R, R) and (S, S) external eliminations of 4-(4-pyridyl)-2,6'(1'H)-dione Spin mixture

外消旋(R,S)與(S,R)或外消旋(R,R)與(S,S) Racemic (R, S) and (S, R) or racemic (R, R) and (S, S)

1H NMR(400MHz,DMSO-d6)：δ ppm 0.75(t,J=7.2Hz,6H),2.62-2.54(m,2H),3.07(d,J=15.6Hz,1H),5.02(d,J=10.0Hz,1H),6.82(s,1H),6.89(d,J=8.4Hz,1H),7.32-7.26(m,1H),7.42(d,J=8.4Hz,2H),7.65(d,J=8.0Hz,2H),10.56(s,1H),11.08(s,1H)。 1 H NMR (400MHz, DMSO- d 6): δ ppm 0.75 (t, J = 7.2Hz, 6H), 2.62-2.54 (m, 2H), 3.07 (d, J = 15.6Hz, 1H), 5.02 (d , J =10.0 Hz, 1H), 6.82 (s, 1H), 6.89 (d, J = 8.4 Hz, 1H), 7.32-7.26 (m, 1H), 7.42 (d, J = 8.4 Hz, 2H), 7.65 (d, J = 8.0 Hz, 2H), 10.56 (s, 1H), 11.08 (s, 1H).

MS：m/z 454.9(M+H)+。 MS: m/z 454.9 (M + H) + .

實例8-9Example 8-9 5-氯-1'-((4-氯苯基)(環丙基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮之(R,R)與(S,S)外消旋混合物或(R,S)與(S,R)外消旋混合物 5-chloro-1'-((4-chlorophenyl)(cyclopropyl)methyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4 a b-pyridine mixture of (R,R) and (S,S) or a racemic mixture of (R,S) and (S,R) of pyridine]-2,6'(1'H)-dione

外消旋(R,R)與(S,S)或外消旋(R,S)與(S,R) Racemic (R, R) and (S, S) or racemic (R, S) and (S, R)

1H NMR(400MHz,DMSO-d6)δ 10.88(br.s.,1H),10.60(br.s.,1H),7.37-7.52(m,4H),7.22-7.36(m,2H),6.87-6.97(m,1H),6.81(s,1H),4.82(d,J=9.54Hz,1H),3.14(d,J=15.89Hz,1H),2.45(s,1H),1.69-1.94(m,1H),0.52-0.83(m,2H),0.28-0.49(m,2H)；MS：m/z 454.0(M+H)+。 1 H NMR (400MHz, DMSO- d6) δ 10.88 (br.s., 1H), 10.60 (br.s., 1H), 7.37-7.52 (m, 4H), 7.22-7.36 (m, 2H), 6.87 -6.97 (m, 1H), 6.81 (s, 1H), 4.82 (d, J = 9.54 Hz, 1H), 3.14 (d, J = 15.89 Hz, 1H), 2.45 (s, 1H), 1.69-1.94 ( m, 1H), 0.52 - 0.83 (m, 2H), 0.28 - 0.49 (m, 2H); MS: m/z 454.0 (M+H) + .

實例8-10Example 8-10 5-氯-1'-((4-氯苯基)(環丙基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮之(R,S)與(S,R)外消旋混合物或(R,R)與(S,S)外消旋混合物 5-chloro-1'-((4-chlorophenyl)(cyclopropyl)methyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3,4 a b-pyridine mixture of (R,S) and (S,R) or a racemic mixture of (R,R) and (S,S) of pyridine]-2,6'(1'H)-dione

外消旋(R,S)與(S,R)或外消旋(R,R)與(S,S) Racemic (R, S) and (S, R) or racemic (R, R) and (S, S)

1H NMR(400MHz,DMSO-d6)δ 10.94(br.s.,1H),10.59(br.s.,1H),7.39-7.53(m,4H),7.28-7.37(m,2H),6.91(d,J=8.31Hz,1H),6.83(s,1H),4.81(d,J=9.54Hz,1H),3.13(d,J=16.14Hz,1H),2.57(s,1H),1.68-1.84(m,1H),0.52-0.78(m,2H),0.24-0.50(m,2H) 1 H NMR (400MHz, DMSO- d6) δ 10.94 (br.s., 1H), 10.59 (br.s., 1H), 7.39-7.53 (m, 4H), 7.28-7.37 (m, 2H), 6.91 (d, J = 8.31 Hz, 1H), 6.83 (s, 1H), 4.81 (d, J = 9.54 Hz, 1H), 3.13 (d, J = 16.14 Hz, 1H), 2.57 (s, 1H), 1.68 -1.84 (m, 1H), 0.52-0.78 (m, 2H), 0.24-0.50 (m, 2H)

MS：m/z 453.9(M+H)+。 MS: m/z 453.9 (M + H) + .

實例8-11Example 8-11 5-氯-1'-(1-(4-氯苯基)-2,2-二甲基丙基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮之(R,R)與(S,S)外消旋混合物或(R,S)與(S,R)外消旋混合物 5-Chloro-1'-(1-(4-chlorophenyl)-2,2-dimethylpropyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazole And [3,4-b]pyridine]-2,6'(1'H)-dione of (R,R) with (S,S) racemic mixture or (R,S) and (S,R Racemic mixture

外消旋(R,R)與(S,S)或外消旋(R,S)與(S,R) Racemic (R, R) and (S, S) or racemic (R, S) and (S, R)

1H NMR(400MHz,DMSO-d 6)δ 11.01(s,1H),10.55(s,1H),7.70-7.63(m,2H),7.47-7.40(m,2H),7.31-7.26(m,2H),6.94-6.88(m,1H),6.85(s,1H),5.36(s,1H),3.12(d,J=16.0Hz,1H),2.44(d,J=16.0Hz,1H),1.23(s,2H),0.93(s,10H)。 1 H NMR (400MHz, DMSO- d 6) δ 11.01 (s, 1H), 10.55 (s, 1H), 7.70-7.63 (m, 2H), 7.47-7.40 (m, 2H), 7.31-7.26 (m, 2H), 6.94-6.88 (m, 1H), 6.85 (s, 1H), 5.36 (s, 1H), 3.12 (d, J = 16.0 Hz, 1H), 2.44 (d, J = 16.0 Hz, 1H), 1.23 (s, 2H), 0.93 (s, 10H).

MS：m/z 470.0(M+H)+。 MS: m/z 470.0 (M + H) + .

實例8-12Example 8-12 5-氯-1'-(1-(4-氯苯基)-2,2-二甲基丙基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮之(R,S)與(S,R)外消旋混合物或(R,R)與(S,S)外消旋混合物 5-Chloro-1'-(1-(4-chlorophenyl)-2,2-dimethylpropyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazole And [3,4-b]pyridine]-2,6'(1'H)-dione (R,S) and (S,R) racemic mixture or (R,R) and (S,S Racemic mixture

外消旋(R,S)與(S,R)或外消旋(R,R)與(S,S) Racemic (R, S) and (S, R) or racemic (R, R) and (S, S)

灰白色固體。1H NMR(400MHz,DMSO-d 6)δ 11.07(s,1H),10.59(s,1H),7.80-7.71(m,2H),7.46-7.38(m,2H),7.30(dd,J=8.2,2.2Hz,2H),7.27(d,J=2.3Hz,2H),6.91(d,J=8.3Hz,2H),6.87(s,1H),5.31(s,1H),3.06(d,J=16.0Hz,1H),2.55(s,1H),1.23(s,1H),0.93(s,10H)。 Off-white solid. 1 H NMR (400MHz, DMSO- d 6) δ 11.07 (s, 1H), 10.59 (s, 1H), 7.80-7.71 (m, 2H), 7.46-7.38 (m, 2H), 7.30 (dd, J = 8.2, 2.2 Hz, 2H), 7.27 (d, J = 2.3 Hz, 2H), 6.91 (d, J = 8.3 Hz, 2H), 6.87 (s, 1H), 5.31 (s, 1H), 3.06 (d, J = 16.0 Hz, 1H), 2.55 (s, 1H), 1.23 (s, 1H), 0.93 (s, 10H).

MS：m/z 470.0(M+H)+. MS: m/z 470.0 (M+H) + .

對於實例8-3至8-12而言，可藉由使用上述實例之程序分離外消旋混合物來製備外消旋混合物之個別(R,R)、(S,S)、(R,S)、(S,R)對映異構體。 For Examples 8-3 to 8-12, the individual (R, R), (S, S), (R, S) of the racemic mixture can be prepared by isolating the racemic mixture using the procedure of the above examples. , (S, R) enantiomer.

藥理學資料Pharmacological data

可藉由使用下文所述任一種檢定來證明本發明化合物之效用。 The utility of the compounds of the invention can be demonstrated by the use of any of the assays described below.

藉由複製子檢定測定登革熱病毒ECDetermination of dengue virus EC by replicon assay 5050

將BHK21 DENV-2複製子細胞以每孔10,000個細胞之密度接種於96孔微量滴定盤中。在37℃下5% CO2培育隔夜之後，以化合物處理細胞。在培育48h之後使用Promega之EndurRen活細胞受質量測螢光素酶活性。在螢光素酶活性量測之後，向每個孔中添加Promega之CellTiter-Glo試劑來測定化合物之細胞毒性。在Clarity發光微量滴定盤讀取器(BioTek)中讀取培養盤。由平均光單位相對於測試化合物濃度之對數繪製劑量反應曲線。藉由非線性回歸分析來計算50%有效濃度(EC50)，即測試化合物使螢光素酶活性減少50%之濃度。 BHK21 DENV-2 replicon cells were seeded at a density of 10,000 cells per well in 96-well microtiter dishes. After overnight incubation at 5% CO2 at 37 °C, the cells were treated with the compound. The EndurRen live cells of Promega were subjected to mass luciferase activity after 48 h of incubation. After luciferase activity assay, Promega's CellTiter-Glo reagent was added to each well to determine the cytotoxicity of the compound. Plates were read in a Clarity luminescent microtiter plate reader (BioTek). A dose response curve is plotted from the log of the average light unit relative to the concentration of the test compound. Nonlinear regression analysis by calculating 50% effective concentration (EC 50), i.e. a test compound of luciferase activity was reduced by 50% of the concentration.

藉由CFI檢定測定登革熱病毒ECDetermination of dengue virus EC by CFI assay 5050

使BHK21細胞胰蛋白酶化且在含有2% FBS之培養基中稀釋至每毫升中2×105個細胞之濃度。以每孔100μl細胞懸浮液(2×104個細胞)施配於一個96孔組織培養盤中(Nunc，96孔透明平底、無菌、Nunclone △表面)。使細胞在37℃、5% CO2下之培養基中生長隔夜，且隨後在37℃、5% CO2下，在不同濃度之測試化合物存在下，由登革熱病毒以MOI(感染多重性)=0.3感染1h。移除病毒接種物，以含有測試化合物之新鮮培養基替代且在37℃、5% CO2下培育48h。將細胞以PBS洗滌一次且用冷甲醇固定10min。以PBS洗滌兩次之後，將固定細胞在室溫下以含有1% FBS及0.05%吐溫20(Tween-20)之PBS阻斷1h。隨後添加一級抗體(4G2)溶液且培育3h。將細胞以PBS洗滌三次，繼而以辣根過氧化物酶(horseradish peroxidase，HRP)結合之抗小鼠IgG培育1h。以PBS洗滌三次之後，向每個孔中添加3,3',5,5'-四甲基聯苯胺(TMB)受質溶液且藉由添加0.5M硫酸終止反應。在450nM下在Tecan Safire II培養盤讀取器中讀取培養盤以用於病毒抗原定量。由平均吸光度相對於測試化合物濃度之對數繪製劑量反應曲線。藉由非線性回歸分析來計算50%有效濃度(EC50)，即測試化合物使病毒E蛋白產量減少50%之濃度。 BHK21 cells were trypsinized and diluted to a concentration of 2 x 10 5 cells per ml in a medium containing 2% FBS. 100 μl of cell suspension (2 × 10 4 cells) per well was dispensed into a 96-well tissue culture dish (Nunc, 96-well clear flat bottom, sterile, Nunclone Δ surface). The cells were grown overnight in medium at 37 ° C, 5% CO 2 , and then infected with dengue virus at MOI (infection multiplicity) = 0.3 at 37 ° C, 5% CO 2 in the presence of different concentrations of test compound. . The virus inoculum was removed, replaced with fresh medium containing the test compound and incubated for 48 h at 37 ° C, 5% CO 2 . The cells were washed once with PBS and fixed with cold methanol for 10 min. After washing twice with PBS, the fixed cells were blocked with PBS containing 1% FBS and 0.05% Tween 20 (Tween-20) for 1 h at room temperature. A primary antibody (4G2) solution was then added and incubated for 3 h. The cells were washed three times with PBS and then incubated with horseradish peroxidase (HRP)-conjugated anti-mouse IgG for 1 h. After washing three times with PBS, a 3,3',5,5'-tetramethylbenzidine (TMB) substrate solution was added to each well and the reaction was stopped by the addition of 0.5 M sulfuric acid. Plates were read in a Tecan Safire II plate reader at 450 nM for viral antigen quantification. A dose response curve is plotted from the log of the average absorbance relative to the concentration of the test compound. Nonlinear regression analysis by calculating 50% effective concentration (EC 50), i.e., reduce the concentration of the test compound of 50% of the E virus protein yield.

下表5中展示本發明之一些代表性化合物之登革熱EC50資料。 Table 5 below show some representative compounds of the present invention dengue The EC 50 data.

以上資料表明本發明之化合物對登革熱病毒具有活性。 The above data indicates that the compounds of the present invention are active against dengue virus.

(R)-5-氯-1'-((5-氯吡啶-2-基)甲基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮(實例1)之活體內小鼠功效研究(R)-5-chloro-1'-((5-chloropyridin-2-yl)methyl)-5',7'-dihydrospiro[porphyrin-3,4'-pyrazolo[3 , in vivo mouse efficacy study of 4-b]pyridine]-2,6'(1'H)-dione (Example 1)

AG129小鼠(缺乏IFN-α/β及IFN-γ受體)由B&K Universal獲得。將小鼠圈養於個別通風之籠中(TechniPlast)且在7週至9週齡時使用。在第0天時小鼠經腹膜內注射2×106PFU之DENV-2(菌株TSV01)(每組6隻動物)。感染後即刻對受感染之小鼠每天兩次以每公斤體重50及100mg或每天一次以每公斤體重100mg經口給予媒劑或以上標題化合物 歷時3天。在感染後第3天，在***(ketamine)及賽拉嗪(xylazine)麻醉下藉由眼球後穿刺獲得血液樣本。將血液樣本收集於含有檸檬酸鈉之管中至最終濃度為0.4%且即刻以6000g離心3min以獲得血漿。隨後，將20μL血漿稀釋於780μL RPMI 1640培養基中且在液氮中快速冷凍以用於溶菌斑檢定分析。簡言之，將BHK-21細胞接種於24孔培養盤(Nunc)中且使其生長至匯合。將細胞層以連續稀釋之病毒樣本培育1h且覆蓋以0.8%甲基纖維素。將培養盤在37℃下培育5天、於10%甲醛中固定且以水中1%之結晶紫染色20min。對溶菌斑以視覺計數。使用單因子ANOVA Tukey-Kramer檢驗來進行比較經不同處理之小鼠組與媒劑對照組之平均值的統計學分析。 AG129 mice (lacking IFN-α/β and IFN-γ receptors) were obtained from B&K Universal. Mice were housed in individual ventilated cages (TechniPlast) and used at 7 weeks to 9 weeks of age. Injected intraperitoneally on day 0 the mice 2 × 10 6 PFU of DENV-2 (strain TSV01) (6 animals per group). Immediately after infection, the infected mice were orally administered with vehicle or the above title compound at a dose of 50 mg and 100 mg/kg of body weight per day for 100 mg/kg of body weight per day for 3 days. On the third day after infection, blood samples were obtained by post-ocular puncture under ketamine and xylazine anesthesia. Blood samples were collected in tubes containing sodium citrate to a final concentration of 0.4% and immediately centrifuged at 6000 g for 3 min to obtain plasma. Subsequently, 20 μL of plasma was diluted in 780 μL of RPMI 1640 medium and snap frozen in liquid nitrogen for plaque assay analysis. Briefly, BHK-21 cells were seeded in 24-well plates (Nunc) and grown to confluence. The cell layer was incubated with serially diluted virus samples for 1 h and covered with 0.8% methylcellulose. The plates were incubated at 37 ° C for 5 days, fixed in 10% formaldehyde and stained with 1% crystal violet in water for 20 min. The plaque is visually counted. Statistical analysis of the mean of the differently treated mice and vehicle controls was performed using a one-way ANOVA Tukey-Kramer test.

以上標題化合物之活體內功效研究的結果呈現於圖1中。其明確表明，此標題化合物在以各種劑量(例如每天兩次50mg/Kg或100mg/Kg，或每天一次100mg/kg)投與小鼠時展現顯著之病毒血症減輕。 The results of the in vivo efficacy studies of the above title compounds are presented in Figure 1. It is clearly shown that this title compound exhibits significant viremia relief when administered to mice at various doses (e.g., 50 mg/Kg or 100 mg/Kg twice daily or 100 mg/kg once daily).

細胞毒性測定Cytotoxicity assay

可使用以下通用方案來測定本發明之代表性化合物的細胞毒性。 The following general scheme can be used to determine the cytotoxicity of representative compounds of the invention.

HepG2細胞中之CCK8細胞毒性檢定CCK8 cytotoxicity assay in HepG2 cells

使HepG2細胞胰蛋白酶化、洗滌、計數且在補充有10%胎牛血清(Fetal bovine serum，FBS)、1%青黴素/鏈黴素、2mM HEPES、1mM丙酮酸鈉、10mM半乳糖及2mM麩醯胺酸之DMEM-W/O葡萄糖中稀釋至每毫升1.6×104個細胞。在透明之384孔組織培養盤中每孔施配25μl含有400個細胞之培養基且在室溫下培育30分鐘。隨後轉移培養盤且置於37℃、5% CO2加濕培育器中隔夜。次日，製備連續稀釋之化合物培養盤且隨後向組織培養孔中施配125nl各種濃度之化合物(200倍稀釋)。隨後將培養盤轉移至37℃、5% CO2加濕培育器中再歷時96 小時。藉由CCK-8檢定來量測細胞毒性。簡言之，將CCK-8在實驗台上部解凍且以生長培養基稀釋2.5倍。隨後向每孔中引入35μl經預稀釋之CCK-8且隨後將培養盤在37℃、5% CO2加濕培育器中再培育3h。由Envision在450nm下讀取吸光度。如前述章節中計算劑量反應曲線。以抑制50%信號之化合物濃度來評估CC50。使用陽性對照(嘌呤黴素)以確保資料之品質。 HepG2 cells were trypsinized, washed, counted and supplemented with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin, 2 mM HEPES, 1 mM sodium pyruvate, 10 mM galactose and 2 mM bran The amino acid DMEM-W/O glucose was diluted to 1.6 × 10 4 cells per ml. 25 μl of medium containing 400 cells was dispensed per well in a transparent 384-well tissue culture dish and incubated at room temperature for 30 minutes. The plates were then transferred and placed overnight at 37 ° C in a 5% CO 2 humidified incubator. On the next day, serially diluted compound culture dishes were prepared and then 125 nl of various concentrations of compound (200-fold dilution) were dispensed into the tissue culture wells. The plates were then transferred to a 37 ° C, 5% CO 2 humidification incubator for a further 96 hours. Cytotoxicity was measured by CCK-8 assay. Briefly, CCK-8 was thawed on top of the bench and diluted 2.5 fold in growth medium. 35 μl of pre-diluted CCK-8 was then introduced into each well and the plates were then incubated for an additional 3 h at 37 ° C in a 5% CO 2 humidified incubator. The absorbance was read by Envision at 450 nm. The dose response curve was calculated as in the previous section. Concentration of compound to inhibit 50% of the evaluated signal CC 50. A positive control (puromycin) is used to ensure the quality of the data.

THP-1細胞中之CCK8細胞毒性檢定CCK8 cytotoxicity assay in THP-1 cells

使用THP-1細胞之4天細胞毒性檢定：對生長於懸浮液中之THP-1細胞計數且在補充有10%胎牛血清(FBS)及1%青黴素/鏈黴素之RPMI-1640培養基中稀釋至每毫升8×104個細胞。向384孔組織培養盤中施配25μl由2000個細胞組成之含有THP-1之培養基且在室溫下預培育30分鐘，繼而在37℃、5% CO2加濕培育器中培育隔夜。次日，製備連續稀釋之化合物培養盤且隨後向組織培養孔中施配125nl各種濃度之化合物(200倍稀釋)。隨後將培養盤轉移至37℃、5% CO2加濕培育器中再歷時96小時。隨後將培養盤轉移至37℃、5% CO2加濕培育器中再歷時96小時。藉由CCK-8檢定來量測細胞毒性。簡言之，將CCK-8在實驗台上部解凍且以生長培養基稀釋2.5倍。隨後向每孔中引入35μl經預稀釋之CCK-8且隨後將培養盤在37℃、5% CO2加濕培育器中再培育3小時。由Envision在450nm下讀取吸光度。如前述章節中計算劑量反應曲線(n=2)。以抑制50%信號之化合物濃度來評估CC50。使用陽性對照(嘌呤黴素)以確保資料之品質。 4-day cytotoxicity assay using THP-1 cells: THP-1 cells grown in suspension were counted and in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin Dilute to 8 x 10 4 cells per ml. 25 μl of a medium containing THP-1 consisting of 2000 cells was administered to a 384-well tissue culture dish and pre-incubated for 30 minutes at room temperature, followed by overnight incubation at 37 ° C in a 5% CO 2 humidified incubator. On the next day, serially diluted compound culture dishes were prepared and then 125 nl of various concentrations of compound (200-fold dilution) were dispensed into the tissue culture wells. The plates were then transferred to a 37 ° C, 5% CO 2 humidification incubator for a further 96 hours. The plates were then transferred to a 37 ° C, 5% CO 2 humidification incubator for a further 96 hours. Cytotoxicity was measured by CCK-8 assay. Briefly, CCK-8 was thawed on top of the bench and diluted 2.5 fold in growth medium. 35 μl of pre-diluted CCK-8 was then introduced into each well and the plates were then incubated for an additional 3 hours at 37 ° C in a 5% CO 2 humidified incubator. The absorbance was read by Envision at 450 nm. The dose response curve (n=2) was calculated as in the previous section. Concentration of compound to inhibit 50% of the evaluated signal CC 50. A positive control (puromycin) is used to ensure the quality of the data.

下表6中展示一些代表性化合物之細胞毒性資料。資料表明，本發明之化合物具有良好選擇性，因為儘管其對受登革熱感染之細胞具有活性，但其對正常細胞無毒。 The cytotoxicity data for some representative compounds are shown in Table 6 below. The data indicate that the compounds of the present invention have good selectivity because, although they are active against dengue infected cells, they are not toxic to normal cells.

表6Table 6

活體內藥物動力學(PK)研究In vivo pharmacokinetic (PK) study

在分別以5及25mg/kg之劑量經靜脈內(i.v.)及經口(p.o.)投與本發明之代表性化合物之後，在CD-1雌性小鼠中測定藥物動力學概況。 經靜脈內及經口使用之調配物分別如下：於NMP：血漿(10：90v/v)中之溶液及於磷酸鹽緩衝液(pH 6.8)中之甲基纖維素(0.5% w/v)吐溫80(0.5% v/v)中之懸浮液。在給藥後0.02(僅靜脈內給藥)、0.08(僅經口給藥)、0.25、0.75、1.5、3、8及24h時收集血液樣本。由LC/MS/MS量測所測試化合物之血漿濃度。藉由使用WinNonLin軟體(Pharsight)之非分割方法計算藥物動力學參數且展示於下表7中。 The pharmacokinetic profile was determined in CD-1 female mice after intravenous (i.v.) and oral (p.o.) administration of representative compounds of the invention at doses of 5 and 25 mg/kg, respectively. The formulations for intravenous and oral use were as follows: solution in NMP: plasma (10:90 v/v) and methylcellulose in phosphate buffer (pH 6.8) (0.5% w/v) Suspension in Tween 80 (0.5% v/v). Blood samples were collected at 0.02 (intravenous administration), 0.08 (oral administration only), 0.25, 0.75, 1.5, 3, 8 and 24 h after administration. The plasma concentration of the test compound was measured by LC/MS/MS. The pharmacokinetic parameters were calculated by using the non-segmentation method of WinNonLin software (Pharsight) and are shown in Table 7 below.

Cmax：測試化合物在血漿中之最大濃度 Cmax: the maximum concentration of test compound in plasma

AUC0-24：外推至無限大之曲線下面積 AUC 0-24 : Extrapolation to the area under the curve of infinity

F%：經口生物可用性 F%: oral bioavailability

作用機制(MoA)：Mechanism of action (MoA):

使用下文所示結構之本發明之代表性化合物(R)-5-氯-1'-(4-氯苄基)-5',7'-二氫螺[吲哚啉-3,4'-吡唑并[3,4-b]吡啶]-2,6'(1'H)-二酮(「測試化合物」)來研究針對登革熱病毒之作用機制。 A representative compound of the present invention (R)-5-chloro-1'-(4-chlorobenzyl)-5',7'-dihydrospiro[porphyrin-3,4'- using the structure shown below. Pyrazolo[3,4-b]pyridine]-2,6'(1'H)-dione ("test compound") was used to study the mechanism of action against dengue virus.

為確定測試化合物是否抑制病毒轉譯及/或RNA合成，使用DENV-2之螢光素酶-報導複製子進行瞬時轉染檢定。複製子經活體外轉錄且轉染至BHK21細胞中，繼而以所示濃度之測試化合物或DMSO進行處理(參見圖2)。監控轉染後各個時間點之螢光素酶活性。在經DMSO處理之細胞(作為對照)中，螢光素酶活性在轉染後4h內達到峰 值且之後逐漸減小。在轉染後16h時，螢光素酶活性再次增加且在轉染後33h時達到峰值。螢光素酶信號之4-h峰代表輸入複製子RNA轉譯，而33-h峰代表RNA合成。在經測試化合物處理之細胞中，測試化合物展示在轉染2h及4h後對螢光素酶信號無作用，而在轉染24h及33h後以劑量-反應性方式遏止螢光素酶活性(參見圖2)。結果表明測試化合物抑制病毒RNA合成。 To determine whether the test compound inhibits viral translation and/or RNA synthesis, transient transfection assays were performed using DENV-2 luciferase-reporter replicons. Replicons were transcribed in vitro and transfected into BHK21 cells, which were then treated with the indicated concentrations of test compound or DMSO (see Figure 2). Luciferase activity was monitored at various time points after transfection. In DMSO-treated cells (as a control), luciferase activity peaked within 4 h after transfection The value then decreases gradually. At 16 h after transfection, luciferase activity increased again and peaked at 33 h after transfection. The 4-h peak of the luciferase signal represents the input replicon RNA translation, while the 33-h peak represents RNA synthesis. In the cells treated with the test compound, the test compound showed no effect on luciferase signal after 2 h and 4 h of transfection, and inhibited luciferase activity in a dose-reactive manner after 24 h and 33 h of transfection (see figure 2). The results indicate that the test compound inhibits viral RNA synthesis.

為識別抗病毒標靶，藉由在遞增濃度之測試化合物存在下培養野生型(WT)病毒來選擇測試化合物抗性DENV。進行六次獨立選擇。對於每次選擇而言，總計進行11次繼代，其中在27nM測試化合物下選擇前3次繼代(P1至P3)，在80nM測試化合物下選擇接下來之4次繼代(P4至P7)且在150nM測試化合物下選擇最後4次繼代(P8至P11)。經由比較來自經模擬處理之感染的病毒效價與來自經測試化合物處理之感染的病毒效價(在感染後72h收集)來檢定來自每次繼代之病毒的抗性。分離能夠在150nM測試化合物(對應於高出約20倍之EC50)存在下生長之P11病毒，使其傳播且檢驗化合物敏感性。對來自六次獨立選擇之P11病毒的完整基因體定序以識別潛在之抗性突變。所有選擇已積聚在7012及/或7013位之核苷酸變化，導致NS4B蛋白中63位之胺基酸Val取代為Ala、Leu、Met、Ser或Thr(參見表8：選定突變病毒之抗性概況)。 To identify antiviral targets, test compound resistant DENV is selected by culturing wild-type (WT) virus in the presence of increasing concentrations of test compound. Make six independent choices. For each selection, a total of 11 passages were performed, with the first 3 passages (P1 to P3) selected under the 27 nM test compound and the next 4 passages (P4 to P7) under the 80 nM test compound. The last 4 passages (P8 to P11) were selected under 150 nM test compound. Resistance from each subcultured virus was assayed by comparing the viral titer from the mock treated infection to the viral titer from the tested compound treated infection (collected 72 h post infection). In the test compounds can be separated 150nM (corresponding to about 20-fold higher than the EC 50) under the presence of P11 viral growth, propagation and so the sensitivity test compound. Complete genome sequencing from six independently selected P11 viruses to identify potential resistance mutations. All selections have accumulated nucleotide changes at positions 7012 and/or 7013, resulting in the substitution of the amino acid Val at position 63 of the NS4B protein for Ala, Leu, Met, Ser or Thr (see Table 8: Resistance of selected mutant viruses) Overview).

為證實V63處之胺基酸取代致使對測試化合物賦予抗性，個別地將NS4B區中之每個突變工程化為DENV之感染性cDNA純系。由工程 化cDNA純系回收之感染性病毒展示對測試化合物處理具有抗性，與親本抗性分離株類似(參見表9：NS4B中殘基63處之胺基酸取代對測試化合物抵抗DENV-2之效能的影響)。 To confirm that the amino acid substitution at V63 confers resistance to the test compound, each of the NS4B regions was individually engineered into an infectious cDNA pure line of DENV. By engineering The infectious virus display recovered from the pure cDNA system is resistant to the treatment of the test compound, similar to the parent resistant isolate (see Table 9: Efficacy of amino acid substitution at residue 63 in NS4B against test compound against DENV-2 Impact).

NS4B之序列對準表明，V63在DENV-2與-3之間絕對保守，但在DENV-1與-4及其他黃病毒之間不同。顯然，突變之63 Ile殘基在DENV-1中作為WT胺基酸存在；突變之63Leu殘基在DENV-4、TBEV及YFV NS4B蛋白中作為WT胺基酸存在；且突變之63Thr殘基在JEV及WNV NS4B蛋白中作為WT胺基酸存在。NS4B之胺基酸63的該等差異可導致測試化合物在抑制DENV-2與-3中之選擇性。將DENV-1 NS4B蛋白之I64殘基工程化為Val，且檢驗DENV-1之抗性表型變為測試化合物。如圖3中所示，I64V突變使DENV-1對測試化合物處理之感受性增加。該等結果表明，NS4B之殘基V63處之突變產生測試化合物抗性。 Sequence alignment of NS4B indicates that V63 is absolutely conserved between DENV-2 and -3, but differs between DENV-1 and -4 and other flaviviruses. Apparently, the mutated 63 Ile residue is present as a WT amino acid in DENV-1; the mutated 63Leu residue is present as a WT amino acid in the DENV-4, TBEV and YFV NS4B proteins; and the mutated 63Thr residue is The JEV and WNV NS4B proteins are present as WT amino acids. These differences in the amino acid 63 of NS4B can result in the selectivity of the test compound in inhibiting DENV-2 and -3. The I64 residue of the DENV-1 NS4B protein was engineered to Val and the resistance phenotype of DENV-1 was tested to become the test compound. As shown in Figure 3, the I64V mutation increased the sensitivity of DENV-1 to the treatment of test compounds. These results indicate that mutations at residue V63 of NS4B result in test compound resistance.

總而言之，如上文所述之突變及遺傳分析表明測試化合物藉由與NS4B蛋白相互作用而抑制DENV複製。因此，本發明之化合物係NS4B抑制劑。 In summary, mutations and genetic analyses as described above indicate that the test compound inhibits DENV replication by interacting with the NS4B protein. Thus, the compounds of the invention are NS4B inhibitors.

Claims (25)

一種式(I)化合物， 其中R1為F、Cl、Br、CF3、OCH3、(C1-C6)烷基或環烷基；R2為H、F、Cl、Br、(C1-C6)烷基或環烷基；或R1與R2連同其所連接之芳族碳原子一起形成稠合1,3-間二氧雜環戊烯基；R3為H、(C1-C6)烷基或環烷基；R4為H、(C1-C6)烷基、環烷基或-C(=O)-NH2；R5為H、(C1-C6)烷基、環烷基或苄基；R6為H、(C1-C6)烷基或環烷基；R7為H、F、Cl或OCH3；R8為F、Cl、Br、CF3、-OCHF2、OCF3、OCH3或(C1-C6)烷基；X1為CH或N；X2為CH或N；X3為CH或N；X4為=O或H2； R1、R2、R3、R4、R5、R6、R7及R8各自視情況獨立地經一或多個獨立R300取代基取代；R300係選自由H、(C1-C6)烷基、環烷基、羥基、胺基、F、Cl、Br、苯基、雜環及雜芳基組成之群；或其醫藥學上可接受之鹽。 a compound of formula (I), Wherein R 1 is F, Cl, Br, CF 3 , OCH 3 , (C 1 -C 6 )alkyl or cycloalkyl; and R 2 is H, F, Cl, Br, (C 1 -C 6 )alkyl Or a cycloalkyl group; or R 1 and R 2 together with the aromatic carbon atom to which they are attached form a fused 1,3-dioxolyl; R 3 is H, (C 1 -C 6 ) alkane Or a cycloalkyl group; R 4 is H, (C 1 -C 6 )alkyl, cycloalkyl or -C(=O)-NH 2 ; R 5 is H, (C 1 -C 6 )alkyl, a cycloalkyl or benzyl group; R 6 is H, (C 1 -C 6 )alkyl or cycloalkyl; R 7 is H, F, Cl or OCH 3 ; R 8 is F, Cl, Br, CF 3 , -OCHF 2 , OCF 3 , OCH 3 or (C 1 -C 6 )alkyl; X 1 is CH or N; X 2 is CH or N; X 3 is CH or N; X 4 is =O or H 2 ; R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 and R 8 are each optionally independently substituted with one or more independent R 300 substituents; R 300 is selected from H, (C 1 a group of -C 6 )alkyl, cycloalkyl, hydroxy, amine, F, Cl, Br, phenyl, heterocyclic and heteroaryl; or a pharmaceutically acceptable salt thereof. 如請求項1之化合物或其醫藥學上可接受之鹽，其中R1為Cl。 The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 is Cl. 如請求項1或2之化合物或其醫藥學上可接受之鹽，其中R2為Cl或F。 A compound according to claim 1 or 2, wherein R 2 is Cl or F, or a pharmaceutically acceptable salt thereof. 如請求項1或2之化合物或其醫藥學上可接受之鹽，其中R3為H或環丙基。 A compound according to claim 1 or 2, wherein R 3 is H or cyclopropyl, or a pharmaceutically acceptable salt thereof. 如請求項1或2之化合物或其醫藥學上可接受之鹽，其中R4為H。 The compound of claim 1 or 2, wherein R 4 is H, or a pharmaceutically acceptable salt thereof. 如請求項1或2之化合物或其醫藥學上可接受之鹽，其中R5為H、甲基或異丙基。 The compound of claim 1 or 2, wherein R 5 is H, methyl or isopropyl, or a pharmaceutically acceptable salt thereof. 如請求項1或2之化合物或其醫藥學上可接受之鹽，其中R6為H或甲基。 The compound of claim 1 or 2, wherein R 6 is H or methyl, or a pharmaceutically acceptable salt thereof. 如請求項1或2之化合物或其醫藥學上可接受之鹽，其中R7為H或Cl。 The compound of claim 1 or 2, wherein R 7 is H or Cl, or a pharmaceutically acceptable salt thereof. 如請求項1或2之化合物或其醫藥學上可接受之鹽，其中R8為Cl、甲基、異丙基或第三丁基。 The compound of claim 1 or 2, wherein R 8 is Cl, methyl, isopropyl or tert-butyl, or a pharmaceutically acceptable salt thereof. 如請求項1或2之化合物或其醫藥學上可接受之鹽，其中X1為CH。 The compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein X 1 is CH. 如請求項1或2之化合物或其醫藥學上可接受之鹽，其中X2為N。 The compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein X 2 is N. 如請求項1或2之化合物或其醫藥學上可接受之鹽，其中X3為CH。 The compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein X 3 is CH. 如請求項1或2之化合物或其醫藥學上可接受之鹽，其中X4為=O。 The compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein X 4 is =0. 如請求項1或2之化合物或其醫藥學上可接受之鹽，其係選自由以下組成之群： A compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, selected from the group consisting of: 如請求項1或2之化合物或其醫藥學上可接受之鹽，其係選自由以下組成之群： A compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, selected from the group consisting of: 如請求項1或2之化合物或其醫藥學上可接受之鹽，其係選自由以下組成之群： A compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, selected from the group consisting of: 如請求項1或2之化合物或其醫藥學上可接受之鹽，其係(R)-5'-氯-3-((S)-1-(4-氯苯基)乙基)-4,6-二氫螺[[1,2,3]***并[4,5-b]吡啶-7,3'-吲哚啉]-2',5(3H)-二酮 A compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, which is (R)-5'-chloro-3-((S)-1-(4-chlorophenyl)ethyl)-4 ,6-Dihydrospiro[[1,2,3]triazolo[4,5-b]pyridine-7,3'-carboline]-2',5(3H)-dione 一種醫藥組合物，其包含治療有效量之如請求項1至17中任一項之化合物或其醫藥學上可接受之鹽及醫藥學上可接受之載劑或賦形劑。 A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient. 如請求項18之醫藥組合物，其進一步包含至少一種其他醫藥劑。 The pharmaceutical composition of claim 18, which further comprises at least one other pharmaceutical agent. 如請求項18或請求項19之醫藥組合物，其中該至少一種其他醫藥劑係西戈斯韋(Celgosivir)。 The pharmaceutical composition of claim 18 or claim 19, wherein the at least one other pharmaceutical agent is Celgosivir. 如請求項1或2之化合物或其醫藥學上可接受之鹽，其係用作藥物。 A compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, for use as a medicament. 一種如請求項1至17中任一項之化合物或其醫藥學上可接受之鹽之用途，其係用於製造用以治療有此需要之個體之由病毒感染 引起之疾病的藥物。 Use of a compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, for the manufacture of a virus for the treatment of an individual in need thereof The drug that causes the disease. 如請求項22之用途，其中該病毒感染係由登革熱病毒(dengue virus)引起。 The use of claim 22, wherein the viral infection is caused by a dengue virus. 如請求項22之用途，其中該個體係人類。 The use of claim 22, wherein the system is human. 如請求項1或2之化合物或其醫藥學上可接受之鹽，其係NS4B抑制劑。 A compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, which is an NS4B inhibitor.
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