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WO2018019297A1 - Isoquinolinone compound and use thereof in preparation of antiviral drugs - Google Patents

Isoquinolinone compound and use thereof in preparation of antiviral drugs Download PDF

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Publication number
WO2018019297A1
WO2018019297A1 PCT/CN2017/094946 CN2017094946W WO2018019297A1 WO 2018019297 A1 WO2018019297 A1 WO 2018019297A1 CN 2017094946 W CN2017094946 W CN 2017094946W WO 2018019297 A1 WO2018019297 A1 WO 2018019297A1
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compound
alkyl
group
mmol
rac
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PCT/CN2017/094946
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French (fr)
Chinese (zh)
Inventor
陈力
翟培彬
邵庆
武进
江涛
李晓闻
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Ginkgo Pharma Co ltd
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Ginkgo Pharma Co ltd
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Priority claimed from CN201610671491.1A external-priority patent/CN107759585A/en
Application filed by Ginkgo Pharma Co ltd filed Critical Ginkgo Pharma Co ltd
Priority to AU2017304952A priority Critical patent/AU2017304952A1/en
Priority to EP17833600.4A priority patent/EP3492467A4/en
Priority to US16/319,777 priority patent/US20190381014A1/en
Priority to CA3031021A priority patent/CA3031021A1/en
Priority to KR1020197002649A priority patent/KR20190022795A/en
Priority to JP2019503692A priority patent/JP2019523261A/en
Publication of WO2018019297A1 publication Critical patent/WO2018019297A1/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4375—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having nitrogen as a ring heteroatom, e.g. quinolizines, naphthyridines, berberine, vincamine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5365—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines ortho- or peri-condensed with heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/54—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
    • A61K31/542—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame ortho- or peri-condensed with heterocyclic ring 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
    • A61P31/20—Antivirals for DNA viruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D455/00—Heterocyclic compounds containing quinolizine ring systems, e.g. emetine alkaloids, protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine
    • C07D455/03—Heterocyclic compounds containing quinolizine ring systems, e.g. emetine alkaloids, protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine containing quinolizine ring systems directly condensed with at least one six-membered carbocyclic ring, e.g. protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine
    • C07D455/04—Heterocyclic compounds containing quinolizine ring systems, e.g. emetine alkaloids, protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine containing quinolizine ring systems directly condensed with at least one six-membered carbocyclic ring, e.g. protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine containing a quinolizine ring system condensed with only one six-membered carbocyclic ring, e.g. julolidine
    • C07D455/06—Heterocyclic compounds containing quinolizine ring systems, e.g. emetine alkaloids, protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine containing quinolizine ring systems directly condensed with at least one six-membered carbocyclic ring, e.g. protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine containing a quinolizine ring system condensed with only one six-membered carbocyclic ring, e.g. julolidine containing benzo [a] quinolizine ring systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/04—Ortho-condensed systems
    • C07D491/044—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • C07D491/048—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring the oxygen-containing ring being five-membered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/04—Ortho-condensed systems
    • C07D491/056—Ortho-condensed systems with two or more oxygen atoms as ring hetero atoms in the oxygen-containing ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D498/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
    • C07D513/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
    • C07D513/04—Ortho-condensed systems

Definitions

  • nucleoside drugs reduce hepatitis B virus by inhibiting the synthesis of viral DNA and have no effect on viral RNA.
  • Nucleoside drugs can not cure hepatitis B and can only suppress the replication of hepatitis B virus. Therefore, a drug that simultaneously inhibits the novel mechanism of action of viral DNA and RNA is required to cure hepatitis B.
  • the newly developed core protein inhibitor can simultaneously inhibit the DNA and RNA of the virus, and the therapeutic effect similar to entecavir can be achieved by using the drug alone.
  • hepatitis B In the design of new drugs for the purpose of curing hepatitis B, the mechanism of the newly designed compounds is the same as that of interferon. It is necessary to reactivate the body's own immune system and rely on its own immune system to identify and remove infected liver cells, thus completely curing hepatitis B. Hepatitis B surface antigen and other viral antigens secreted by hepatocytes from patients with chronic hepatitis B interfere with the immune system through signal transduction systems, block the recognition of viruses by immune cells and further limit their antiviral function. In addition, persistent and excessive hepatitis B surface antigens can inactivate the immune system, delete T-cells, and perform functional damage.
  • nucleoside drugs can not reduce hepatitis B surface antigen, and it is necessary to design a new mechanism of action, combined with potent nucleoside drugs, and effectively remove hepatitis B surface antigen and viral DNA in the blood, activate and restore itself. Immune function, which may eventually cure hepatitis B.
  • An object of the present invention is to provide novel isoquinolinone compounds which have an extremely strong activity for inhibiting hepatitis B DNA and a potent inhibitory activity against hepatitis B surface antigen.
  • the structure of this class of compounds will block the pathway of P450 oxidation, increase the bioavailability of the compound, and reduce the toxicity of the compound.
  • These highly active compounds will be combined with nucleoside compounds and TLR7 agonists, which may significantly improve the therapeutic effect and cure rate of hepatitis B in the clinic.
  • the present invention adopts the following technical solutions:
  • R 2 is selected from the group consisting of halogen, C 1-3 alkoxy, deuterated C 1-3 alkoxy, C 1-6 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy , C 4-8 cycloheteroalkyl C 1-6 alkyl, halo C 1-3 alkyloxy, halo C 3-6 cycloalkyl, C 3-6 cycloalkyl C 1-6 alkane a group or R 2 and R 3 are bonded by a carbon atom to form a ring;
  • R 5 , R 5 ' independently selected from hydrogen, hydrazine, halogen, methyl, methoxy or R 5 , R 5 'forms a carbocyclic or heterocyclic ring; or R 5 , R 6 form a carbocyclic or heterocyclic ring ring;
  • W is N or CR 7 , wherein R 7 is selected from the group consisting of hydrogen, hydrazine, hydroxy, halogen, C 1-3 alkyl, C 1-6 alkoxy, C 3-6 cycloalkyloxy, ester , carboxyl or cyano;
  • the present invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising the isoquinolinone compound of the formula (I) according to the present invention, a stereoisomer, a pharmaceutically acceptable salt, a solvate or a crystal thereof, and a pharmaceutically acceptable An acceptable carrier or excipient.
  • the invention provides novel isoquinolinone compounds, which have strong inhibition of hepatitis B DNA activity, EC 50 can be less than 5 nanomolar, and have strong activity for inhibiting hepatitis B surface antigen, and EC 50 is About 10 nanomoles.
  • such compounds have excellent pharmacokinetic properties.
  • the compounds of the invention will block the pathway of P450 oxidation, increase the bioavailability of the compounds, and reduce the toxicity of the compounds.
  • These highly active compounds will be combined with nucleoside compounds and TLR7 agonists, which may significantly improve the therapeutic effect and cure rate of hepatitis B in the clinic.
  • hydrocarbyl refers to a straight, branched or cyclic, saturated or unsaturated substituent consisting essentially of carbon and hydrogen. It is preferably 1 to 20 carbon atoms, more preferably 1 to 12 carbon atoms.
  • alkyl refers to a straight, branched or cyclic saturated hydrocarbon group.
  • cycloalkyl refers to a saturated and/or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon group.
  • a single ring can include from 3 to 10 carbon atoms.
  • monocyclic cycloalkane groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl and the like.
  • Polycyclic cycloalkyl groups include spiro, fused, and bridged cycloalkyl groups.
  • the cycloalkyl group includes an unsubstituted group and a substituent.
  • aryl refers to a 6-10 membered all carbon monocyclic or polycyclic aromatic group including phenyl, naphthyl, biphenyl, and the like.
  • the aryl group can be substituted and unsubstituted.
  • ⁇ is an isotope of hydrogen with an atomic mass twice that of the latter and a stronger binding to carbon.
  • Deuterated “and” ⁇ means that hydrogen is replaced with deuterium at the specified position.
  • a “deuterated substituent” is a substituent in which at least one hydrogen is replaced by deuterium enriched at a specified percentage.
  • heterocyclyl refers to a cyclic group containing at least one heteroatom, wherein the heteroatoms are nitrogen, oxygen, sulfur, sulfur, and the like.
  • the heterocyclic group includes a monoheterocyclic group and a polyheterocyclic group.
  • DIBAL-H Diisobutylaluminum hydride
  • PhN(OTf) 2 N-phenyl bis(trifluoromethanesulfonimide)
  • Compound 26a-rac Compound 22i-rac (186 mg, 0.52 mmol) was dissolved in 20 mL of DMF, and Intermediate 8d (100mg, 0.52mmol) and potassium carbonate (142mg, 1.04mmol). The mixture was heated and stirred at 85 ° C for 3 hours, cooled to room temperature, then added with 60 mL of water, and then extracted with ethyl acetate (50 mL ⁇ 3). The organic layer was combined, washed with saturated sodium chloride, dried and concentrated The next step is to react.
  • Compound 28c-rac Compound 28b-rac (3.40 g, 10.9 mmol) was dissolved in 50 mL of dioxane, and toluene (2.51 g, 54.5 mmol). After cooling to room temperature, 50 mL of a saturated aqueous solution of sodium hydrogencarbonate was added, and then ethyl acetate (50 mL ⁇ 3), and the organic layer was combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate and evaporated. In the next step.
  • Compound 28f-rac Compound 28e-rac (6.40 g, 13.8 mmol) was dissolved in 40 mL of ethylene glycol dimethyl ether, and tetrachlorophenylhydrazine (3.40 g, 13.8 mmol) was added, and the mixture was stirred under heating at 55 ° C for 3 hours. The mixture was added to a silica gel and purified by column chromatography to give a product 3.20 g. Yield: 50.2%.
  • Compound 30a-rac Compound 28g-rac (85mg, 0.23mmol), compound 6c (89mg, 0.46mmol) and potassium carbonate (63mg, 0.46mmol) were added to 5mL DMF, replaced with nitrogen three times, heated and stirred at 90 °C Reaction for 3 hours. After completion of the reaction, the mixture was diluted with EtOAc (EtOAc m.
  • Compound 31a-rac Compound 28g-rac (65mg, 0.18mmol), compound 7e (55mg, 0.26mmol) and potassium carbonate (50mg, 0.36mmol) were added to 5mL DMF, replaced with nitrogen three times, heated and stirred at 90 °C Reaction for 3 hours. After completion of the reaction, the mixture was diluted with EtOAc (EtOAc m.
  • Compound 52f-rac Compound 52e (0.63 g, 1.84 mmol) was dissolved in 5 mL of dioxane, and 1.0 mL of formic acid and 0.5 mL of triethyl orthoformate were added, and the mixture was stirred under reflux for 48 hours. After completion of the reaction, the solvent was evaporated to dryness.
  • Compound 52h-rac Compound 52g-rac (0.58g, 1.64mmol) and ethyl 2-ethoxymethyleneacetate (0.92g, 4.92mmol) were dissolved in 6mL of ethanol, 2mL water was added, and the reaction was stirred and heated under reflux. After 48 hours, after completion of the reaction, the reaction solvent was dried to give 1.17 g of crude product.
  • Compound 52i-rac Compound 52h-rac (1.17 g, 2.36 mmol) and tetrachlorophenylhydrazine (0.35 g, 1.42 mmol) were dissolved in 15 mL of ethylene glycol dimethyl ether and stirred under reflux for 3 hours. The reaction solvent was spin-dried, and the product was purified by silica gel column chromatography. Yield: 40.9%.
  • Compound 54a-rac Compound 52j-rac (100 mg, 0.25 mmol), Compound 6c (97 mg, 0.50 mmol) and potassium carbonate (103 mg, 0.75 mmol) After adding to 5 mL of DMF, the nitrogen gas was replaced three times, and the reaction was stirred under heating at 90 ° C for 5 hours. After completion of the reaction, the mixture was diluted with EtOAc (EtOAc)EtOAc.
  • Compound I-54-rac Compound 54a-rac (110 mg, 0.22 mmol) was dissolved in 3 mL of tetrahydrofuran, 40 mg of sodium hydroxide and 1 mL of water were added, and the reaction was stirred at 35 ° C for 2 hours. After completion of the reaction, the pH was adjusted to about 2-3 with 1N hydrochloric acid, and extracted with dichloromethane (20 mL ⁇ 3). The organic phase was combined and dried over anhydrous sodium sulfate. Yield: 26.1%.
  • Compound 56a-rac Compound 52j-rac (100 mg, 0.25 mmol), compound 8d (96 mg, 0.50 mmol) and potassium carbonate (103 mg, 0.75 mmol) were added to 5 mL of DMF, replaced with nitrogen three times and heated and stirred at 90 °C. Reaction for 5 hours. After completion of the reaction, it was diluted with water and extracted with ethyl acetate (20 mL ⁇ 3). The organic phase was combined and dried over anhydrous sodium sulfate. Yield: 40.0%.
  • Compound I-56-rac Compound 56a-rac (60 mg, 0.1 mmol) was dissolved in 4 mL of tetrahydrofuran, 25 mg of sodium hydroxide and 1 mL of water were added, and the reaction was stirred at 35 ° C for 2 hours. After the completion of the reaction, the pH was adjusted to about 2-3 with 1N hydrochloric acid, and extracted with dichloromethane (20 mL ⁇ 3). The organic phase was combined, dried over anhydrous sodium sulfate, Yield: 84.7%.
  • the preparations for administration were freshly prepared on the day of administration, and samples were taken for the determination of the actual concentration of the preparation for administration.
  • Group A SD rats were given a single intravenous injection (IV) with a dose of 2 mg ⁇ kg -1
  • group B SD rats were given a single dose (PO) with a dose of 10 mg ⁇ kg -1
  • 0.15 mL of blood was collected from the jugular vein before administration and 5 minutes after administration (intravenous group only), 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours and 24 hours.
  • All whole blood samples were centrifuged (5500 rpm) for 10 minutes, and the plasma was separated and stored in a refrigerator at -30 to -10 °C.
  • the concentration of the test compound in the plasma of SD rats was determined by LC-MS/MS analysis.
  • the corresponding pharmacokinetic parameters were calculated using a non-compartmental model in Pharsight Phoenix 7.0. See Tables 2a and 2b for the results.

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Abstract

Disclosed is an isoquinolinone compound as shown in formula (I) or a stereoisomer, pharmaceutically acceptable salt, solvate or crystal thereof, and a preparation thereof and a use thereof in the preparation of drugs for treating or preventing viral infectious diseases caused by the hepatitis B virus (HBV) and other viruses, in particular, a use thereof in drugs for HBV Surface antigen inhibitors and HBV DNA production inhibitors. The compound has a significant activity for inhibiting the hepatitis B surface antigen and hepatitis B DNA, and it is possible to significantly improve the probability of curing hepatitis B by administration of same in combination with nucleoside drugs or other drugs in the future, and same has good clinical application prospects.

Description

异喹啉酮类化合物及作为抗病毒药物的应用Isoquinolinone compounds and their use as antiviral drugs 技术领域Technical field

本发明属于医药化学领域，具体涉及一类新型异喹啉酮类化合物或其立体异构体，含有前述异喹啉酮类化合物或其立体异构体的药物组合物以及它们作为抗病毒药物的用途，特别是作为乙肝表面抗原抑制剂(HBV Surface antigen inhibitors)和乙肝DNA抑制剂(HBV DNA production inhibitors)的药物用于治疗和/或预防感染乙肝病毒的用途，特别是指这类化合物和TLR7抑制剂以及核苷类药物可以作为治愈乙肝的药物组合物的用途。The invention belongs to the field of medical chemistry, and particularly relates to a novel class of isoquinolinone compounds or stereoisomers thereof, a pharmaceutical composition containing the aforementioned isoquinolinone compounds or stereoisomers thereof and their use as antiviral drugs. Use, especially as a drug for hepatitis B surface antigen inhibitors and HBV DNA production inhibitors, for the treatment and/or prevention of infection with hepatitis B virus, especially for such compounds and TLR7 Inhibitors and nucleoside drugs can be used as a pharmaceutical composition for curing hepatitis B.

背景技术Background technique

慢性乙型肝炎在全球大约有3.5亿感染，在2011年有78万人因为乙型肝炎死亡，其中中国的乙肝患者占全球乙肝患者总数的约三分之一。中国目前每年用于乙肝治疗的费用超过1000亿，是全球最大的乙肝药物市场。尽管乙肝疫苗已经广泛应用，中国的乙肝病人仍以每年平均250万人左右的速度迅速增长，美国的乙肝病人也以15.4％的速度快速增长。乙肝病毒的携带者大约有25％的病人转变为慢性乙肝，10-30％的慢性乙肝发展为肝硬化或肝癌。慢性乙肝是导致肝硬化的主要因素之一。There are about 350 million infections in chronic hepatitis B worldwide. In 2011, 780,000 people died of hepatitis B. Among them, hepatitis B patients in China accounted for about one-third of the total number of hepatitis B patients worldwide. China currently spends more than 100 billion yuan a year on hepatitis B treatment, making it the world's largest market for hepatitis B drugs. Although hepatitis B vaccine has been widely used, hepatitis B patients in China are growing at an average rate of 2.5 million per year, and hepatitis B patients in the United States are also growing at a rate of 15.4%. About 25% of carriers of hepatitis B virus are converted to chronic hepatitis B, and 10-30% of chronic hepatitis B develops into cirrhosis or liver cancer. Chronic hepatitis B is one of the main factors leading to cirrhosis.

目前FDA批准的治疗乙肝的药物有7个，他们分别是干扰素-α，聚乙二醇化干扰素-α，拉米夫定，恩替卡韦，替比夫定，阿德福韦酯和替诺福韦酯。TAF(tenofovir alafenamide fumarate，替诺福韦艾拉酚胺富马酸)已经完成三期临床，等待FDA批准。所有的这些药物都不能有效的治愈乙肝，需要长期服药。干扰素类药物通过刺激人体的免疫系统抑制病毒的DNA和RNA，使用干扰素耐药性较少，有一定的乙肝表面抗原消失和血清转换，缺点是应答率低，需要注射和严重的副作用。拉米夫定和替比夫定容易产生耐药性，不能长期服用。例如服用拉米夫定在第一年有20％的病人产生耐药性，第二年有70％的病人产生耐药性。阿德福韦酯由于耐受性和不良反应逐渐退出一线药物。目前WHO推荐治疗乙肝的一线药物为替诺福韦酯(TDF)和恩替卡韦。特别是替诺福韦酯在连续服用5年没有发现耐药性，并且病毒清除率达95％-100％，副作用小。所有这些核苷类药物都是通过抑制病毒DNA的合成来降低乙肝病毒，对于病毒的RNA没有作用。核苷类药物也不能治愈乙肝，只能压制乙肝病毒的复制。因此需要同时抑制病毒DNA和RNA的全新作用机理的药物治愈乙肝。最新开发的核心蛋白抑制剂可以同时抑制病毒的DNA和RNA，单独用药可以达到和恩替卡韦相似的治疗效果。其中NVR-3-778是一种有效的衣壳抑制剂，但是没有乙肝表面抗原(HBsAg)消失的数据。从现有的数据分析，衣壳抑制剂也不能治愈乙肝，因为这类化合物不能有效的作用于乙肝的cccDNA，也没有数据支持衣壳抑制剂可以缩短cccDNA的半衰期，二期临床研究的衣壳抑制剂需要和核苷类药物或干扰素合用。Currently, there are 7 drugs approved by the FDA for the treatment of hepatitis B. They are interferon-α, pegylated interferon-α, lamivudine, entecavir, telbivudine, adefovir dipivoxil and tenofovir. Wei ester. TAF (tenofovir alafenamide fumarate, tenofovir alafenamide fumarate) has completed Phase III clinical trials, awaiting FDA approval. All of these drugs are not effective in curing hepatitis B and require long-term medication. Interferon drugs inhibit the virus's DNA and RNA by stimulating the body's immune system. They use less interferon resistance, have certain hepatitis B surface antigen disappearance and seroconversion. The disadvantage is that the response rate is low, requiring injection and serious side effects. Lamivudine and telbivudine are prone to drug resistance and cannot be taken for long periods of time. For example, in the first year, 20% of patients taking lamivudine developed drug resistance, and in the second year, 70% of patients developed drug resistance. Adefovir dipivoxil gradually withdraws from first-line drugs due to tolerance and adverse reactions. The first line of WHO recommended for the treatment of hepatitis B is tenofovir (TDF) and entecavir. In particular, tenofovir disoproxil did not find drug resistance for 5 consecutive years, and the virus clearance rate was 95%-100%, and the side effects were small. All of these nucleoside drugs reduce hepatitis B virus by inhibiting the synthesis of viral DNA and have no effect on viral RNA. Nucleoside drugs can not cure hepatitis B and can only suppress the replication of hepatitis B virus. Therefore, a drug that simultaneously inhibits the novel mechanism of action of viral DNA and RNA is required to cure hepatitis B. The newly developed core protein inhibitor can simultaneously inhibit the DNA and RNA of the virus, and the therapeutic effect similar to entecavir can be achieved by using the drug alone. Among them, NVR-3-778 is an effective capsid inhibitor, but there is no data on the disappearance of hepatitis B surface antigen (HBsAg). From the existing data analysis, capsid inhibitors can not cure hepatitis B because these compounds can not effectively act on cccDNA of hepatitis B, and there is no data to support capsid inhibitors can shorten the half-life of cccDNA, the capsid of the second phase clinical study Inhibitors need to be combined with nucleoside drugs or interferons.

以治愈乙肝为目的的新药设计，新设计化合物的机理要求和干扰素一样，需要重新激活人体自身的免疫系统，依靠自身的免疫系统识别和清除感染的肝细胞，从而彻底治愈乙肝。慢性乙肝病人的肝细胞分泌的乙肝表面抗原和其他病毒抗原，通过信号转导系统，干扰免疫系统，阻断免疫细胞对病毒的识别和进一步限制其发挥抗病毒功能。另外持续和过多的乙肝表面抗原可以使免疫系统钝化，T-细胞缺失和进行功能损伤。乙肝表面抗原还可以直接压制免疫细胞对病毒的清除功能。基于以上的原因，开发针对抑制乙肝表面抗原分泌的药物可以有效的恢复免疫细胞的功能，减轻免疫系统的压力，使免疫系统识别并清除感染的肝细胞，达到直接治愈乙肝的目的。另外，乙肝表面抗原减少也是慢性乙肝好转的生物指标，乙肝表面抗原消失和血清转换表明乙肝已经功能性治愈。目前核苷类药物不能减少乙肝表面抗原，必需设计新的作用机理的药物，和强效的核苷类药物联用，同时有效的清除乙肝表面抗原和血液中的病毒DNA，激活并恢复自身的免疫功能，从而最终可能治愈乙肝。In the design of new drugs for the purpose of curing hepatitis B, the mechanism of the newly designed compounds is the same as that of interferon. It is necessary to reactivate the body's own immune system and rely on its own immune system to identify and remove infected liver cells, thus completely curing hepatitis B. Hepatitis B surface antigen and other viral antigens secreted by hepatocytes from patients with chronic hepatitis B interfere with the immune system through signal transduction systems, block the recognition of viruses by immune cells and further limit their antiviral function. In addition, persistent and excessive hepatitis B surface antigens can inactivate the immune system, delete T-cells, and perform functional damage. Hepatitis B surface antigen can also directly suppress the clearance function of immune cells by viruses. Based on the above reasons, the development of drugs for inhibiting the secretion of hepatitis B surface antigen can effectively restore the function of immune cells, reduce the pressure of the immune system, enable the immune system to recognize and remove infected liver cells, and achieve the goal of directly curing hepatitis B. In addition, the reduction of hepatitis B surface antigen is also a biological indicator of the improvement of chronic hepatitis B. The disappearance of hepatitis B surface antigen and seroconversion indicate that hepatitis B has been functionally cured. At present, nucleoside drugs can not reduce hepatitis B surface antigen, and it is necessary to design a new mechanism of action, combined with potent nucleoside drugs, and effectively remove hepatitis B surface antigen and viral DNA in the blood, activate and restore itself. Immune function, which may eventually cure hepatitis B.

发明内容Summary of the invention

本发明的目的在于提供新型的异喹啉酮类化合物，该类化合物有极强的抑制乙肝DNA的活性，并具有强效的抑制乙肝表面抗原的活性。此外，该类化合物的结构将阻断P450氧化的途径，提高化合物的生物利用度，降低化合物的毒性。这些高活性的化合物将和核苷类化合物以及TLR7激动剂联合用药，在临床上可能显著提高乙肝的治疗效果和治愈率。SUMMARY OF THE INVENTION An object of the present invention is to provide novel isoquinolinone compounds which have an extremely strong activity for inhibiting hepatitis B DNA and a potent inhibitory activity against hepatitis B surface antigen. In addition, the structure of this class of compounds will block the pathway of P450 oxidation, increase the bioavailability of the compound, and reduce the toxicity of the compound. These highly active compounds will be combined with nucleoside compounds and TLR7 agonists, which may significantly improve the therapeutic effect and cure rate of hepatitis B in the clinic.

为实现上述目的，本发明采用如下技术方案：To achieve the above object, the present invention adopts the following technical solutions:

一种式(I)所示的异喹啉酮类化合物或其立体异构体、可药用盐、溶剂化物或结晶， An isoquinolinone compound of the formula (I) or a stereoisomer thereof, a pharmaceutically acceptable salt, a solvate or a crystal thereof,

Figure PCTCN2017094946-appb-000001
Figure PCTCN2017094946-appb-000001

其中：among them:

(1)R1选自H、氘、C1-6烷基、氰基、卤素、羧基、酯基、C3-6环烷基、C4-8杂环烷基、卤代C1-6烷基或C6-10芳基；(1) R 1 is selected from the group consisting of H, hydrazine, C 1-6 alkyl, cyano, halogen, carboxyl, ester, C 3-6 cycloalkyl, C 4-8 heterocycloalkyl, halogen C 1- 6 alkyl or C 6-10 aryl;

(2)R2选自卤素、C1-3烷氧基、氘代C1-3烷氧基、C1-6烷基、C3-6环烷基、C3-6环烷基氧基、C4-8环杂烷基C1-6烷基、卤代C1-3烷基氧基、卤代C3-6环烷基、C3-6环烷基C1-6烷基或R2和R3以碳原子连接形成环；(2) R 2 is selected from the group consisting of halogen, C 1-3 alkoxy, deuterated C 1-3 alkoxy, C 1-6 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy , C 4-8 cycloheteroalkyl C 1-6 alkyl, halo C 1-3 alkyloxy, halo C 3-6 cycloalkyl, C 3-6 cycloalkyl C 1-6 alkane a group or R 2 and R 3 are bonded by a carbon atom to form a ring;

(3)R3为(a)具有环结构和/或不饱和键的C4-12烃基，该C4-12烃基的氢原子未被取代，或为选自氘、卤素、氰基、羟基、巯基中的一个或多个取代基取代，且该C4-12烃基未被杂原子间断，或被O，S，NH，C＝O，C＝S，O＝S＝O中的一个或多个所间断，所述杂原子选自氧、硫或氮；或者，(b)R2和R3以碳原子连接形成环；(3) R 3 is (a) a C 4-12 hydrocarbon group having a ring structure and/or an unsaturated bond, the hydrogen atom of the C 4-12 hydrocarbon group is unsubstituted, or is selected from the group consisting of ruthenium, halogen, cyano, and hydroxy Substituting one or more substituents in the fluorenyl group, and the C 4-12 hydrocarbon group is not interrupted by a hetero atom, or is one of O, S, NH, C=O, C=S, O=S=O or a plurality of discontinuities, the hetero atom is selected from oxygen, sulfur or nitrogen; or, (b) R 2 and R 3 are bonded by a carbon atom to form a ring;

(4)R4选自氢、氘、卤素、氰基、酯基或C1-3烷基；(4) R 4 is selected from the group consisting of hydrogen, hydrazine, halogen, cyano, ester or C 1-3 alkyl;

(5)R5，R5′独立的选自氢、氘、卤素、甲基、甲氧基或R5，R5′形成碳环或杂环；或R5，R6形成碳环或杂环；(5) R 5 , R 5 ' independently selected from hydrogen, hydrazine, halogen, methyl, methoxy or R 5 , R 5 'forms a carbocyclic or heterocyclic ring; or R 5 , R 6 form a carbocyclic or heterocyclic ring ring;

(6)M为CH或N；(6) M is CH or N;

(7)R6选自C1-6烷基、C1-6烷氧C1-6烷基、羟基C1-6烷基、芳基、卤代C1-6烷基或C3-6环烷基C1-6烷基；(7) R 6 is selected from C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, hydroxy C 1-6 alkyl, aryl, halogen C 1-6 alkyl or C 3- 6 cycloalkyl C 1-6 alkyl;

(8)W为N或CR7，其中R7选自氢、氘、羟基、卤素、C1-3烷基、C1-6烷氧基、C3-6环烷基氧基、酯基、羧基或氰基；(8) W is N or CR 7 , wherein R 7 is selected from the group consisting of hydrogen, hydrazine, hydroxy, halogen, C 1-3 alkyl, C 1-6 alkoxy, C 3-6 cycloalkyloxy, ester , carboxyl or cyano;

(9)R8选自羧基、酯基、C1-6烷基、C3-6环烷基、C1-6烷基炔基或C3-6环烷基炔基，其中所述酯基的烷基部分选自C1-6烷基、C3-8环烷基、C3-8环烷基炔基、C1-6烷基炔基、苄基、C1-6烷基C(O)O-C1-3烷基、C1-6烷基-OC(O)O-C1-3烷基。(9) R 8 is selected from a carboxyl group, an ester group, a C 1-6 alkyl group, a C 3-6 cycloalkyl group, a C 1-6 alkyl alkynyl group or a C 3-6 cycloalkyl alkynyl group, wherein the ester The alkyl moiety of the group is selected from the group consisting of C 1-6 alkyl, C 3-8 cycloalkyl, C 3-8 cycloalkyl alkynyl, C 1-6 alkyl alkynyl, benzyl, C 1-6 alkyl C(O)OC 1-3 alkyl, C 1-6 alkyl-OC(O)OC 1-3 alkyl.

根据本发明，所述被O，S，NH，C＝O，C＝S，O＝S＝O中的一个或多个所间断的烃基是指烃基的相邻接的二个碳原子或该烃基与其所连接的碳原子之间被这些原子或基团所间断，在满足有机化合物的成键规则的前提下，对于间断的位置没有特别限制，被多个原子间断时，这些间隔原子或基团可以是相邻位置或间隔开的，当间隔原子或基团为多个时，它们可以是多个相同的原子或基团，也可以是不同的原子或基团，当间隔原子为多个不同的原子或基团且它们处于相邻位置时，可形成新的间隔基团，例如COO(酯基)、酰胺基(CONH)、SO2NH(磺酰胺基)等。例如，被O，S，NH，C＝O，C＝S，O＝S＝O中的一个所间断的丙基可以是例如OCH2CH2CH3；CH2OCH2CH3；CH2SCH2CH3；CH2NHCH2CH3，CH2COCH2CH3，CH2COCH2CH3，CH2SO2CH2CH3；例如被O，S，NH，C＝O，C＝S，O＝S＝O中的二个所间断的丙基可以是例如CH2COOCH2CH3，CH2COCH2OCH3，CH2CONHCH2CH3，CH2C＝OCHNHCH3，CH2SO2NHCH2CH3等。According to the present invention, the one or more interrupted hydrocarbon groups of O, S, NH, C=O, C=S, and O=S=O refer to two adjacent carbon atoms of the hydrocarbon group or The hydrocarbon group and the carbon atom to which it is attached are interrupted by these atoms or groups. Under the premise that the bonding rules of the organic compound are satisfied, there is no particular limitation on the position of the discontinuity. When the atoms are interrupted by a plurality of atoms, the spacer atoms or groups are The clusters may be adjacent or spaced apart. When the spacer atoms or groups are plural, they may be a plurality of identical atoms or groups, or may be different atoms or groups, when the spacer atoms are multiple When different atoms or groups are present and they are in adjacent positions, new spacer groups such as COO (ester group), amide group (CONH), SO 2 NH (sulfonamide group) and the like can be formed. For example, a propyl interrupted by one of O, S, NH, C=O, C=S, O=S=O may be, for example, OCH 2 CH 2 CH 3 ; CH 2 OCH 2 CH 3 ; CH 2 SCH 2 CH 3 ; CH 2 NHCH 2 CH 3 , CH 2 COCH 2 CH 3 , CH 2 COCH 2 CH 3 , CH 2 SO 2 CH 2 CH 3 ; for example by O, S, NH, C=O, C=S, The two interrupted propyl groups in O=S=O may be, for example, CH 2 COOCH 2 CH 3 , CH 2 COCH 2 OCH 3 , CH 2 CONHCH 2 CH 3 , CH 2 C=OCHNHCH 3 , CH 2 SO 2 NHCH 2 CH 3 and so on.

进一步地，所述(a)中，所述环结构优选为3～8元环，更优选为3～6元环；所述不饱和键可以为双键或三键。Further, in the above (a), the ring structure is preferably a 3- to 8-membered ring, more preferably a 3- to 6-membered ring; and the unsaturated bond may be a double bond or a triple bond.

优选地，所述(a)中，所述的环结构为饱和环。Preferably, in the above (a), the ring structure is a saturated ring.

优选地，所述(a)中，所述环结构、不饱和键的个数分别为1～2个。Preferably, in the above (a), the number of the ring structure and the unsaturated bond is 1 to 2, respectively.

优选地，所述(a)具有一个3～8元饱和碳环或3～8元饱和杂环、至少一个杂原子或至少一个双键或三键。Preferably, the (a) has a 3 to 8 membered saturated carbocyclic ring or a 3 to 8 membered saturated heterocyclic ring, at least one hetero atom or at least one double or triple bond.

进一步优选地，所述(a)中，所述环结构、不饱和键以及杂原子中的至少二者同时存在。Further preferably, in the above (a), at least two of the ring structure, the unsaturated bond, and the hetero atom are simultaneously present.

根据本发明的一个具体且优选方面，所述(a)为满足如下任意一项所述条件的基团：According to a particular and preferred aspect of the invention, the (a) is a group which satisfies the conditions described in any one of the following:

a1)同时具有所述的环结构和碳碳不饱和键，且环结构和碳碳不饱和键分别有且只有一个；A1) having both the ring structure and the carbon-carbon unsaturated bond, and having one and only one ring structure and carbon-carbon unsaturated bond;

a2)同时具有所述的环结构和1～3个杂原子，且所述杂原子中至少一个为氧，且该氧原子通过单键与式(I)中苯环连接；A2) having both the ring structure and 1 to 3 hetero atoms, and at least one of the hetero atoms is oxygen, and the oxygen atom is bonded to the benzene ring in the formula (I) through a single bond;

a3)同时具有不饱和键和1～3个杂原子，其中不饱和键为碳碳双键，碳碳三键或者碳氧双键。优选地，不饱和键为碳碳双键，碳碳三键时，它们的一端通过单键与式(I)中苯环连接。A3) has both an unsaturated bond and 1 to 3 hetero atoms, wherein the unsaturated bond is a carbon-carbon double bond, a carbon-carbon triple bond or a carbon-oxygen double bond. Preferably, the unsaturated bond is a carbon-carbon double bond, and at the carbon-carbon triple bond, one end thereof is bonded to the benzene ring in the formula (I) through a single bond.

根据本发明的一个具体方面，R3为C5-11双环烷基；C3-6环烷基炔基；C3-6环烷基烯基；C1-3烷氧C1-6烷基炔基；C1-3烷氧C1-6烷基烯基；C4-8环杂烷基；或者， According to a particular aspect of the invention, R 3 is C 5-11 bicycloalkyl; C 3-6 cycloalkylalkynyl; C 3-6 cycloalkylalkenyl; C 1-3 alkoxy C 1-6 alkane Alkynyl; C 1-3 alkoxy C 1-6 alkylalkenyl; C 4-8 cycloheteroalkyl; or

R3为RA-O-，其中RA选自C3-8环烷基；C5-11双环烷基；氘代C1-6烷基；C4-8环杂烷基；C1-6烷基羰基C1-6烷基；氘代C1-3烷氧基C1-6烷基；C1-3烷氧基C3-8环烷基；C1-3烷氧基C3-8环烷基C1-6烷基；C3-8杂环烷基；C1-3烷氧基C1-6烷基，其中烷基被C3-8环烷烃或C4-8杂环烷烃取代，杂环烷烃的杂原子选自氧、硫或氮；当RA选自C1-3烷氧基C1-6烷基时，R5、R5′独立选自氘、氟、氯、羟基、氰基，且W为N或CR7，其中R7选自氘、氟、氯、羟基、氰基。R 3 is R A -O-, wherein R A is selected from C 3-8 cycloalkyl; C 5-11 bicycloalkyl; deuterated C 1-6 alkyl; C 4-8 cycloheteroalkyl; C 1 -6 alkylcarbonyl C 1-6 alkyl; deuterated C 1-3 alkoxy C 1-6 alkyl; C 1-3 alkoxy C 3-8 cycloalkyl; C 1-3 alkoxy C 3-8 cycloalkyl C 1-6 alkyl; C 3-8 heterocycloalkyl; C 1-3 alkoxy C 1-6 alkyl, wherein alkyl is C 3-8 cycloalkane or C 4 a heterocyclic alkane substituted, the hetero atom of the heterocycloalkane is selected from oxygen, sulfur or nitrogen; when R A is selected from C 1-3 alkoxy C 1-6 alkyl, R 5 , R 5 ' are independently selected from Anthracene, fluorine, chlorine, hydroxyl, cyano, and W is N or CR 7 , wherein R 7 is selected from the group consisting of hydrazine, fluorine, chlorine, hydroxyl, and cyano.

根据本发明的一个优选方面，所述R3选自C3-8环烷氧基、C3-8杂环烷氧基、C1-3烷氧C3-8环烷氧基、C1-3烷氧C3-8环烷基C1-6烷氧基、C3-8杂环烷基、C1-3烷氧C2-9烯基、C1-3烷氧C2-9炔基、C3-8环烷基C2-9烯基、C3-8环烷基C2-9炔基。According to a preferred aspect of the invention, said R 3 is selected from the group consisting of C 3-8 cycloalkoxy, C 3-8 heterocycloalkoxy, C 1-3 alkoxy C 3-8 cycloalkoxy, C 1 -3 alkoxy C 3-8 cycloalkyl C 1-6 alkoxy, C 3-8 heterocycloalkyl, C 1-3 alkoxy C 2-9 alkenyl, C 1-3 alkoxy C 2- 9 alkynyl, C 3-8 cycloalkyl C 2-9 alkenyl, C 3-8 cycloalkyl C 2-9 alkynyl.

根据本发明，所述R２优选为选自C1-3烷氧基、卤素、C3-6环烷基、苄基。According to the invention, the R 2 is preferably selected from the group consisting of C 1-3 alkoxy, halogen, C 3-6 cycloalkyl, benzyl.

根据本发明的一个具体方面，R6选自甲基、乙基、异丙基、丁基、异丁基、甲氧基甲基、甲氧基乙基、甲氧基异丙基、甲氧基丁基、甲氧基异丁基、乙氧基甲基、乙氧基乙基、乙氧基异丙基、乙氧基丁基、乙氧基异丁基、羟基甲基、羟基乙基、羟基异丙基、羟基丁基、羟基异丁基。According to a particular aspect of the invention, R 6 is selected from the group consisting of methyl, ethyl, isopropyl, butyl, isobutyl, methoxymethyl, methoxyethyl, methoxyisopropyl, methoxy Butyl, methoxyisobutyl, ethoxymethyl, ethoxyethyl, ethoxy isopropyl, ethoxybutyl, ethoxyisobutyl, hydroxymethyl, hydroxyethyl , hydroxyisopropyl, hydroxybutyl, hydroxyisobutyl.

根据本发明，除去活泼氢之外，其余所有的氢原子都可以分别独立地被氘取代。According to the present invention, all of the hydrogen atoms except the active hydrogen can be independently substituted by hydrazine.

根据本发明，典型的异喹啉酮类化合物如下列化合物：According to the invention, typical isoquinolinone compounds are as follows:

Figure PCTCN2017094946-appb-000002
Figure PCTCN2017094946-appb-000002

Figure PCTCN2017094946-appb-000003
Figure PCTCN2017094946-appb-000003

Figure PCTCN2017094946-appb-000004
Figure PCTCN2017094946-appb-000004

本发明还提供一种制备本发明所述的式(I)所示的异喹啉酮类化合物、其立体异构体、可药用盐、溶剂化物或结晶的中间体，该中间体如下式(II)所示：The present invention also provides an intermediate of the isoquinolinone compound represented by the formula (I), a stereoisomer, a pharmaceutically acceptable salt, a solvate or a crystal thereof, which is prepared according to the invention. (II):

Figure PCTCN2017094946-appb-000005
式(II)中，所述R1、R２、R4、R5、R5’、R6、R8、W和N的定义同前。
Figure PCTCN2017094946-appb-000005
In the formula (II), R 1 , R 2 , R 4 , R 5 , R 5 ', R 6 , R 8 , W and N have the same meanings as defined above.

根据本发明的一个具体优选方面，式(II)所示的中间体为化合物10或其异构体或消旋体According to a particularly preferred aspect of the invention, the intermediate of formula (II) is compound 10 or an isomer or racemate thereof

Figure PCTCN2017094946-appb-000006
Figure PCTCN2017094946-appb-000006

本发明还进一步提供一种本发明所述的式(I)所示的异喹啉酮类化合物、其立体异构体、可药用盐、溶剂化物或结晶(以下统称本发明化合物)的制备方法，其采用如下式(II)所示的中间体： The present invention still further provides a preparation of an isoquinolinone compound represented by the formula (I), a stereoisomer thereof, a pharmaceutically acceptable salt, a solvate or a crystal thereof (hereinafter collectively referred to as a compound of the present invention). The method employs an intermediate represented by the following formula (II):

Figure PCTCN2017094946-appb-000007
式(II)中，所述R1、R2、R4、R5、R5’、R6、R8、W和N的定义同前。
Figure PCTCN2017094946-appb-000007
In the formula (II), R 1 , R 2 , R 4 , R 5 , R 5 ', R 6 , R 8 , W and N have the same meanings as defined above.

进一步地，所述方法包括使式(II)所示的中间体与RAOH、RAOMs或RABr反应，其中，当反应物为RAOH，所述反应采用Mitsunobu反应，在脱水剂为三苯基膦和/或偶氮二甲酸二乙酯存在下进行；当反应物为RAOMs或RABr，所述反应为SN2反应，在碱为碳酸钾和/或碳酸铯以及催化量KI存在下进行。Further, the method comprises reacting an intermediate represented by the formula (II) with R A OH, R A OMs or R A Br , wherein when the reactant is R A OH, the reaction is carried out by using a Mitsunobu reaction. The agent is carried out in the presence of triphenylphosphine and/or diethyl azodicarboxylate; when the reactant is R A OMs or R A Br, the reaction is SN2 reaction, and the base is potassium carbonate and/or cesium carbonate and The catalytic amount is carried out in the presence of KI.

根据本发明的一个具体方面，式(II)所示的中间体为化合物10或其异构体或消旋体According to a particular aspect of the invention, the intermediate of formula (II) is compound 10 or an isomer or racemate thereof

Figure PCTCN2017094946-appb-000008
Figure PCTCN2017094946-appb-000008

本发明还提供一种药物组合物，其含有本发明所述的式(I)所示的异喹啉酮类化合物、其立体异构体、可药用盐、溶剂化物或结晶，以及药学上可接受的载体或赋形剂。The present invention also provides a pharmaceutical composition comprising the isoquinolinone compound of the formula (I) according to the present invention, a stereoisomer, a pharmaceutically acceptable salt, a solvate or a crystal thereof, and a pharmaceutically acceptable An acceptable carrier or excipient.

优选地，所述药物组合物为抗病毒药物组合物，其中还包含一种或多种治疗剂，所述治疗剂治选自以下组成的群：核苷类药物、利巴韦林、干扰素、HBV衣壳抑制剂(capsid inhibitor)、cccDNA形成抑制剂、cccDNA表观遗传修饰剂或乙肝RNAi药物、TLR7激动剂。Preferably, the pharmaceutical composition is an antiviral pharmaceutical composition, further comprising one or more therapeutic agents, the therapeutic agent being selected from the group consisting of nucleoside drugs, ribavirin, interferon , HBV capsid inhibitor, cccDNA formation inhibitor, cccDNA epigenetic modifier or hepatitis B RNAi drug, TLR7 agonist.

本发明还涉及本发明所述的式(I)所示的异喹啉酮类化合物、其立体异构体、可药用盐、溶剂化物或结晶或其与一种或多种选自核苷类药物、利巴韦林、干扰素、HBV衣壳抑制剂(capsid inhibitor)、cccDNA形成抑制剂、cccDNA表观遗传修饰剂或乙肝RNAi药物、TLR7激动剂的治疗剂的组合在制备预防和/或治疗病毒感染疾病药物，和/或乙肝表面抗原抑制剂(HBV Surface antigen inhibitors)和乙肝DNA抑制剂(HBV DNA production inhibitors)药物中的应用，所述病毒感染包括HBV或HDV的感染。The present invention also relates to the isoquinolinone compound of the formula (I) according to the present invention, a stereoisomer, a pharmaceutically acceptable salt, a solvate or a crystal thereof or one or more selected from the group consisting of nucleosides Combination of drugs, ribavirin, interferon, HBV capsid inhibitor, cccDNA formation inhibitor, cccDNA epigenetic modifier or hepatitis B RNAi drug, TLR7 agonist therapeutic agent in the preparation of prophylaxis and / Or in the treatment of a virus-infected disease drug, and/or a hepatitis B surface antigen inhibitor (HBV), and a hepatitis B virus production inhibitor (HBV DNA production inhibitors), the virus infection including an infection of HBV or HDV.

本发明同时还提供所述药物组合物在制备用于治疗或预防乙肝和乙肝病毒感染药物中的应用以及采用所述药物组合物预防或减缓乙肝和乙肝病毒感染患者疾病的方法。The present invention also provides a method of using the pharmaceutical composition for the preparation of a medicament for treating or preventing hepatitis B and hepatitis B virus infection, and a method for preventing or slowing the disease of a patient infected with hepatitis B and hepatitis B virus using the pharmaceutical composition.

根据本发明的药物组合物，其中本发明所述化合物优选以治疗有效量存在。A pharmaceutical composition according to the invention wherein the compound of the invention is preferably present in a therapeutically effective amount.

上述药物组合物中药学上可接受的载体，如药学上可接受的稀释剂、赋型剂、填充剂、粘合剂、崩解剂、吸收促进剂、表面活性剂、润滑剂、香味剂、甜味剂等。A pharmaceutically acceptable carrier, such as a pharmaceutically acceptable diluent, excipient, filler, binder, disintegrant, absorption enhancer, surfactant, lubricant, fragrance, Sweeteners, etc.

以本发明化合物为活性成分制备的药物可以是片剂、粉剂、胶囊、粒剂、口服液以及注射制剂等多种形式。药物组合物的剂型优选为片剂、胶囊或针剂。The drug prepared by using the compound of the present invention as an active ingredient may be in various forms such as a tablet, a powder, a capsule, a granule, an oral solution, and an injection preparation. The dosage form of the pharmaceutical composition is preferably a tablet, capsule or injection.

上述各种剂型的药物均可以按药学领域的常规方法制备。The above various dosage forms of the drug can be prepared by a conventional method in the pharmaceutical field.

本发明还提供本发明化合物在制备预防或治疗病毒感染疾病中的用途，优选其中病毒感染疾病为HBV病毒感染。The invention also provides the use of a compound of the invention in the manufacture of a medicament for the prevention or treatment of a viral infection, preferably wherein the viral infection is an HBV viral infection.

本发明的药物组合物组成可以由下配比构成：The composition of the pharmaceutical composition of the present invention may be composed of the following ratio:

Figure PCTCN2017094946-appb-000009
Figure PCTCN2017094946-appb-000009

由于以上技术方案的实施，本发明与现有技术相比存在如下优势：Due to the implementation of the above technical solutions, the present invention has the following advantages compared with the prior art:

本发明提供了新型的异喹啉酮类化合物，该类化合物有极强的抑制乙肝DNA的活性，EC50可低于5纳摩尔，并具有强效的抑制乙肝表面抗原的活性，EC50在10纳摩尔左右。此外，该类化合物具有优异的药代动力学特性。进一步地，本发明化合物将阻断P450氧化的途径，提高化合物的生物利用度，降低化合物的毒性。这些高活性的化合物将和核苷类化合物以及TLR7激动剂联合用药，在临床上可能显著提高乙肝的治疗效果和治愈率。The invention provides novel isoquinolinone compounds, which have strong inhibition of hepatitis B DNA activity, EC 50 can be less than 5 nanomolar, and have strong activity for inhibiting hepatitis B surface antigen, and EC 50 is About 10 nanomoles. In addition, such compounds have excellent pharmacokinetic properties. Further, the compounds of the invention will block the pathway of P450 oxidation, increase the bioavailability of the compounds, and reduce the toxicity of the compounds. These highly active compounds will be combined with nucleoside compounds and TLR7 agonists, which may significantly improve the therapeutic effect and cure rate of hepatitis B in the clinic.

术语定义Definition of Terms

除非另外定义，本文使用的所有技术和科学术语具有与本发明所属领域的普通技术人员通常理解的相同的含义。术语“立体异构体”是指由分子中原子在空间上排列方式不同所产生的异构体。包括顺反异构体、对映异构体和构象异构体。所有立体异构体均属于本发明的范围。本发明的化合物可以为单独立体异构体或其它异构体的混合例如外消旋体，或者所有其它立体异构体的混合。Unless otherwise defined, all technical and scientific terms used herein have the same meaning meaning meaning The term "stereoisomer" refers to an isomer produced by the different arrangement of atoms in a molecule in space. These include cis and trans isomers, enantiomers and conformational isomers. All stereoisomers are within the scope of the invention. The compounds of the invention may be a single stereoisomer or a mixture of other isomers such as a racemate, or a mixture of all other stereoisomers.

术语“盐”是指本发明所述的化合物与酸形成的药学上可接受的盐，所述的酸可以是有机酸或无机酸，具体可选自：磷酸、硫酸、盐酸、氢溴酸、柠檬酸、马来酸、丙二酸、扁桃酸、琥珀酸、富马酸、醋酸、乳酸、硝酸、磺酸、对甲苯磺酸、苹果酸、甲烷磺酸或其类似物。The term "salt" refers to a pharmaceutically acceptable salt of a compound of the invention formed with an acid, which may be an organic or inorganic acid, specifically selected from the group consisting of phosphoric acid, sulfuric acid, hydrochloric acid, hydrobromic acid, Citric acid, maleic acid, malonic acid, mandelic acid, succinic acid, fumaric acid, acetic acid, lactic acid, nitric acid, sulfonic acid, p-toluenesulfonic acid, malic acid, methanesulfonic acid or the like.

术语“溶剂化物”是指通过与溶剂分子配位形成固态或液态的配合物的本发明化合物的形式。水合物是溶剂合物的特殊形式，其中与水发生配位。在本发明范围内，溶剂合物优选是水合物。The term "solvate" refers to a form of a compound of the invention that forms a solid or liquid complex by coordination with a solvent molecule. Hydrates are a special form of solvates in which coordination occurs with water. Within the scope of the invention, the solvate is preferably a hydrate.

术语“结晶”是指本发明所述的化合物形成的各种固体形态，包括晶型、无定形。The term "crystalline" refers to the various solid forms formed by the compounds described herein, including crystalline forms, amorphous forms.

术语“烃基”是指直链、支链或环状的饱和或不饱和主要由碳和氢构成的取代基。优选1-20个碳原子，更优选1-12个碳原子。术语“烷基”是指直链、支链或环状的饱和烃基。烷基具体包括甲基、乙基、正丙基、异丙基、环丙基、正丁基、异丁基、叔丁基、环丁基、正戊基、异戊基、新戊基、环己基、正己基、异己基、2，2，-甲基丁基和2，3-二甲基丁基、16-烷基、18-烷基。术语“C1-20烷基”是指含有1-20个碳原子的直链、支链或环状的饱和烃基。烷基包括取代的和没有取代的烷基。当烷基被取代时，取代基可以在任何可使用的连接点上取代，取代基可以是单取代或多取代。取代基独立的选自烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、氘、卤素、硫醇、羟基、硝基、羧基、酯基、氰基，环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、氧代，在命名时取代基通常置于烷基之前，例如C1-3烷氧基C3-8环烷基C1-6烷基指C1-6烷基，其被C3-8环烷基取代，而该C3-8环烷基又为C1-3烷氧基取代，举例：甲氧基环丁基甲基的结构式为：The term "hydrocarbyl" refers to a straight, branched or cyclic, saturated or unsaturated substituent consisting essentially of carbon and hydrogen. It is preferably 1 to 20 carbon atoms, more preferably 1 to 12 carbon atoms. The term "alkyl" refers to a straight, branched or cyclic saturated hydrocarbon group. The alkyl group specifically includes methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, tert-butyl, cyclobutyl, n-pentyl, isopentyl, neopentyl, Cyclohexyl, n-hexyl, isohexyl, 2,2,-methylbutyl and 2,3-dimethylbutyl, 16-alkyl, 18-alkyl. The term "C 1-20 alkyl" refers to a straight, branched or cyclic saturated hydrocarbon group containing from 1 to 20 carbon atoms. Alkyl groups include substituted and unsubstituted alkyl groups. When an alkyl group is substituted, the substituent may be substituted at any available point of attachment, and the substituent may be mono- or poly-substituted. The substituents are independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, anthracene, halogen, thiol, hydroxy, nitro, carboxy, ester, cyano, cycloalkyl , aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, oxo, where the substituent is usually placed before the alkyl group, for example C 1-3 alkoxy C 3- The 8 -cycloalkyl C 1-6 alkyl group means a C 1-6 alkyl group which is substituted by a C 3-8 cycloalkyl group, and the C 3-8 cycloalkyl group is further substituted by a C 1-3 alkoxy group, for example The structural formula of methoxycyclobutylmethyl is:

Figure PCTCN2017094946-appb-000010
Figure PCTCN2017094946-appb-000010

术语“烯基”和“炔基”分别是指直链、支链或环状的含有双键和三键的不饱和烃基，优选2-20个碳原子，更优选2-12个碳原子。烯基、炔基包括取代的和没有取代的烯基、炔基。当被取代时，取代基可以在任何可使用的连接点上取代，取代基可以是单取代或多取代。取代基独立的选自烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、氘、卤素、硫醇、羟基、硝基、羧基、酯基、氰基，环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、氧代，在命名时取代基通常置于烯基、炔基之前。The terms "alkenyl" and "alkynyl" mean, respectively, a straight-chain, branched or cyclic unsaturated hydrocarbon group containing a double bond and a triple bond, preferably 2 to 20 carbon atoms, more preferably 2 to 12 carbon atoms. Alkenyl, alkynyl groups include substituted and unsubstituted alkenyl, alkynyl groups. When substituted, the substituent may be substituted at any available point of attachment, and the substituent may be mono- or poly-substituted. The substituents are independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, anthracene, halogen, thiol, hydroxy, nitro, carboxy, ester, cyano, cycloalkyl , aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, oxo, the substituent is usually placed before the alkenyl, alkynyl group.

术语“环烷基”指饱和和/或部分不饱和单环或多环环烃基。单环可包括3-10个碳原子。单环环烷烃基的非限制实例包括环丙基、环丁基、环戊基、环戊烯基、环己基、环己烯基、环己二烯基、环庚基等。多环环烷基包括螺环、稠环和桥环的环烷基。环烷基包括无取代基和含有取代基。取代基选自一个或多个取代基团，包括但不仅限于以下基团，独立的选自烷基、环烷基、烷氧基、卤素、羧基、酯基、氨基、酰胺基、羟基、氰基、硝基、芳基、杂芳基。The term "cycloalkyl" refers to a saturated and/or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon group. A single ring can include from 3 to 10 carbon atoms. Non-limiting examples of monocyclic cycloalkane groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl and the like. Polycyclic cycloalkyl groups include spiro, fused, and bridged cycloalkyl groups. The cycloalkyl group includes an unsubstituted group and a substituent. The substituent is selected from one or more substituent groups including, but not limited to, the following groups, independently selected from the group consisting of alkyl, cycloalkyl, alkoxy, halogen, carboxyl, ester, amino, amide, hydroxy, cyano Base, nitro, aryl, heteroaryl.

术语“芳基”指6-10元全碳单环或多环的芳香基团，包括苯基，奈基，联苯基等。芳基可以是取代的和未取代的。取代基独立的选自烷基、环烷基(环丙烷基、环丁烷基和环戊烷基等)、烯基、炔基、叠氮、氨基、氘、烷氧基、烷硫基、烷基氨基，卤素、硫醇、羟基，硝基、杂环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基、烷基硅基等。The term "aryl" refers to a 6-10 membered all carbon monocyclic or polycyclic aromatic group including phenyl, naphthyl, biphenyl, and the like. The aryl group can be substituted and unsubstituted. The substituents are independently selected from the group consisting of alkyl, cycloalkyl (cyclopropyl, cyclobutane, cyclopentyl, etc.), alkenyl, alkynyl, azide, amino, hydrazine, alkoxy, alkylthio, Alkylamino, halogen, thiol, hydroxy, nitro, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, alkyl Silicon based and so on.

术语“杂芳基”指包含1-10个杂原子的杂芳香体系的基团。杂原子包括氧，硫，氮，磷等。其中单杂环基包括但不限于呋喃、噻吩、吡咯、噻唑、咪唑、1，2，3-三氮唑、1，2，4-三氮唑、1，2，3-噻二唑，噁唑、1，2，4-噁二唑、1，3，4-噁二唑、吡啶、嘧啶、哒嗪、吡嗪、四氢呋喃、四氢吡咯、哌啶、哌嗪、吗啉、异噁唑啉等。稠杂环基包括但不限于喹啉、异喹啉、吲哚、苯并呋喃、苯并噻吩、嘌呤、吖啶、咔唑、芴、色烯酮、芴酮、喹喔啉、3，4-二氢萘酮、二苯并呋喃、氢化二苯并呋喃、苯并噁唑基等。杂芳基可以是取代的和未取代的。取代基独立的选自取代基独立的选自烷基、环烷基(环丙烷基、环丁烷基和环戊烷基等)、烯基、炔基、叠氮、氨基、氘、烷 氧基、烷硫基、烷基氨基，卤素、硫醇、羟基，硝基、杂环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基、烷基硅基等。The term "heteroaryl" refers to a radical of a heteroaromatic system containing from 1 to 10 heteroatoms. Heteroatoms include oxygen, sulfur, nitrogen, phosphorus, and the like. Wherein the monoheterocyclic group includes, but is not limited to, furan, thiophene, pyrrole, thiazole, imidazole, 1,2,3-triazole, 1,2,4-triazole, 1,2,3-thiadiazole, cacao Azole, 1,2,4-oxadiazole, 1,3,4-oxadiazole, pyridine, pyrimidine, pyridazine, pyrazine, tetrahydrofuran, tetrahydropyrrole, piperidine, piperazine, morpholine, isoxazole Porphyrin and the like. The fused heterocyclic group includes, but is not limited to, quinoline, isoquinoline, indole, benzofuran, benzothiophene, anthracene, acridine, oxazole, indole, chromenone, anthrone, quinoxaline, 3, 4 - dihydronaphthalenone, dibenzofuran, hydrogenated dibenzofuran, benzoxazolyl, and the like. Heteroaryl groups can be substituted and unsubstituted. The substituents are independently selected from the group consisting of alkyl, cycloalkyl (cyclopropyl, cyclobutyl, and cyclopentyl, etc.), alkenyl, alkynyl, azide, amino, anthracene, and alkane. Oxyl, alkylthio, alkylamino, halogen, thiol, hydroxy, nitro, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, hetero A cycloalkylthio group, an alkylsilyl group or the like.

术语“卤素”是指氟、氯、溴、碘，优选为氟、氯、溴。The term "halogen" means fluoro, chloro, bromo, iodo, preferably fluoro, chloro, bromo.

术语“氘”是氢的同位素，原子质量是后者的2倍，与碳的结合更强。氘化“和”氘“表示氢在指定位置被替换为氘。一个“氘化的取代基”是取代基，其中至少一个氢被以指定的百分比富集的氘取代。The term "氘" is an isotope of hydrogen with an atomic mass twice that of the latter and a stronger binding to carbon. Deuterated "and" 氘 means that hydrogen is replaced with deuterium at the specified position. A "deuterated substituent" is a substituent in which at least one hydrogen is replaced by deuterium enriched at a specified percentage.

术语“卤代烷基”是指至少被一个卤素原子取代的烷基。The term "haloalkyl" refers to an alkyl group substituted with at least one halogen atom.

术语“杂环基”是指至少含有一个杂原子的环状基团，其中杂原子为氮、氧、硫、硫等。杂环基包括单杂环基和多杂环基。The term "heterocyclyl" refers to a cyclic group containing at least one heteroatom, wherein the heteroatoms are nitrogen, oxygen, sulfur, sulfur, and the like. The heterocyclic group includes a monoheterocyclic group and a polyheterocyclic group.

具体实施方式Detailed ways

以下实施例可以使本专业技术人员更全面地理解本发明，但不以任何方式限制本发明。所有化合物的结构均经1H NMR或MS所确定。The following examples are intended to provide a more complete understanding of the invention, and are not intended to limit the invention. The structure of all compounds was determined by 1 H NMR or MS.

实施例中用到的化合物名称缩写如下：The compound names used in the examples are abbreviated as follows:

DCM：二氯甲烷DCM: dichloromethane

EtOAc：乙酸乙酯EtOAc: ethyl acetate

THF：四氢呋喃THF: tetrahydrofuran

DME：乙二醇二甲醚DME: ethylene glycol dimethyl ether

Dioxane：1，4-二氧六环Dioxane: 1,4-dioxane

Pd2(dba)3：三(二亚苄基丙酮)二钯Pd 2 (dba) 3 : tris(dibenzylideneacetone) dipalladium

Xantphos：4，5-双二苯基膦-9，9-二甲基氧杂葸Xantphos: 4,5-bis-diphenylphosphino-9,9-dimethyloxazepine

tBuONa：叔丁醇钠tBuONa: sodium tert-butoxide

POCl3：三氯氧磷POCl 3 : phosphorus oxychloride

NH4OAc：醋酸铵NH 4 OAc: ammonium acetate

NaBH3CN：氰基硼氢化钠NaBH 3 CN: sodium cyanoborohydride

p-chloranil：四氯苯醌P-chloranil: tetrachlorophenylhydrazine

MsCl：甲基磺酰氯MsCl: Methanesulfonyl chloride

Et3N：三乙胺Et 3 N: triethylamine

BnBr：苄溴BnBr: benzyl bromide

DIBAL-H：二异丁基氢化铝DIBAL-H: Diisobutylaluminum hydride

PPh3：三苯基膦PPh 3 : triphenylphosphine

DEAD：偶氮二甲酸二乙酯DEAD: diethyl azodicarboxylate

PhN(OTf)2：N-苯基双(三氟甲烷磺酸亚胺)PhN(OTf) 2 : N-phenyl bis(trifluoromethanesulfonimide)

B(OMe)3：硼酸三甲酯B(OMe) 3 : Trimethyl borate

nBuLi：正丁基锂nBuLi: n-butyllithium

Pd(dppf)Cl2：[1，1′-双(二苯基膦基)二茂铁]二氯化钯Pd(dppf)Cl 2 :[1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride

下面结合具体实施例，对本发明做进一步的说明：The present invention will be further described below in conjunction with specific embodiments:

化合物10的制备： Preparation of Compound 10:

Figure PCTCN2017094946-appb-000011
Figure PCTCN2017094946-appb-000011

化合物2的制备：在氮气保护下，化合物1(350g，1.72mol)，苄溴(352g，2.06mol)和碳酸钾(368g，2.66mol)的混合物在乙腈(4.5L)和丙酮(4.3L)的混合溶剂中回流18小时。反应液冷却，过滤，滤液浓缩后残余物加入乙酸乙酯稀释，有机相依次用水和饱和食盐水洗涤，无水硫酸钠干燥，过滤，浓缩，柱层析纯化得到化合物2(437g，86.4％)。1HNMR(CDCl3，300MHz)δ：7.33-7.48(m，5H)，7.04-7.08(m，2H)，7.77(s，1H)，5.12(s，2H)，3.86(s，3H)。Preparation of compound 2: a mixture of compound 1 (350 g, 1.72 mol), benzyl bromide (352 g, 2.06 mol) and potassium carbonate (368 g, 2.66 mol) in acetonitrile (4.5 L) and acetone (4.3 L) under nitrogen. The mixture was refluxed for 18 hours. The reaction mixture was cooled, filtered, and the filtrate was evaporated. EtOAcjjjjjjjjj . 1 H NMR (CDCl 3 , 300 MHz) δ: 7.33-7.48 (m, 5H), 7.04-7.08 (m, 2H), 7.77 (s, 1H), 5.12 (s, 2H), 3.86 (s, 3H).

化合物3的制备：化合物2(330g，1.12mol)溶于四氢呋喃(2.8L)，然后加入3-甲基-2-丁酮(145g，1.69mol)，Pd2(dba)3(三(二亚苄基丙酮)二钯，10g，11.2mmol)，Xantphos(4，5-双二苯基膦-9，9-二甲基氧杂葸，19g，33.6mmol)和叔丁醇钠(162g，1.69mol)。反应体系用氮气置换三次，然后在氮气保护下于55℃反应6小时。旋干，剩余物中加入水(800mL)和乙酸乙酯(1000mL)并搅拌30分钟。静置分层，水相用乙酸乙酯(500mL×2)萃取。合并有机相，用饱和食盐水洗涤。无水硫酸钠干燥，过滤，滤液旋干后柱层析纯化得到化合物3(187g，55.6％)。1HNMR(CDCl3，300MHz)δ：7.46-7.28(m，5H)，6.88-6.76(m，3H)，5.15(s，2H)，3.89(s，3H)，3.63(s，2H)，2.68-2.63(m，1H)，1.05(d，J＝6.9Hz，6H)。Preparation of Compound 3: Compound 2 (330 g, 1.12 mol) was dissolved in tetrahydrofuran (2.8 L), then 3-methyl-2-butanone (145 g, 1.69 mol), Pd 2 (dba) 3 (three (2) Benzylacetone) dipalladium, 10 g, 11.2 mmol), Xantphos (4,5-bisdiphenylphosphino-9,9-dimethyloxaxan, 19 g, 33.6 mmol) and sodium tert-butoxide (162 g, 1.69) Mol). The reaction system was replaced with nitrogen three times, and then reacted at 55 ° C for 6 hours under a nitrogen atmosphere. It was dried to dryness, and water (800 mL) and ethyl acetate (1000 mL) were added and stirred for 30 minutes. The layers were allowed to stand and the aqueous phase was extracted with ethyl acetate (500 mL×2). The organic phases were combined and washed with brine. The residue was dried over anhydrous sodium sulfate, filtered, and then evaporated to dryness. 1 H NMR (CDCl 3 , 300 MHz) δ: 7.46-7.28 (m, 5H), 6.88-6.76 (m, 3H), 5.15 (s, 2H), 3.89 (s, 3H), 3.63 (s, 2H), 2.68 -2.63 (m, 1H), 1.05 (d, J = 6.9 Hz, 6H).

化合物4的制备：往甲醇(1.4L)中加入化合物3(180g，0.6mol)和醋酸铵(465g，6.0mol)并用冰水浴冷却，然后往里面分批加入氰基硼氢化钠(30g，0.48mol)，于室温下搅拌10分钟，然后50℃搅拌21小时。冷却后加入水(490mL)和10N氢氧化钠溶液(120mL，1.2mol)，室温下搅拌1小时。然后用二氯甲烷(500mL×3)萃取，合并有机相，用饱和食盐水洗涤，无水硫酸钠干燥后旋干得到化合物4(176g，97.4％)。1HNMR(CDCl3，300MHz)δ：7.46-7.28(m，5H)，6.86-6.76(m，3H)，5.16(s，2H)，3.88(s，3H)，2.76-2.72(m，2H)，2.37-2.29(m，1H)，1.63-1.54(m，3H)，0.94(d，J＝6.9Hz，6H)。Preparation of compound 4: To a solution of methanol (1.4 L), compound 3 (180 g, 0.6 mol) and ammonium acetate (465 g, 6.0 mol) were added and cooled in an ice water bath, and then sodium cyanoborohydride (30 g, 0.48) was added portionwise. Mol), stirred at room temperature for 10 minutes and then stirred at 50 ° C for 21 hours. After cooling, water (490 mL) and a 10N sodium hydroxide solution (120 mL, 1.2 mol) were added and stirred at room temperature for 1 hour. Then, it was extracted with dichloromethane (500 mL × 3), and the organic phase was combined, washed with saturated brine, dried over anhydrous sodium sulfate and then evaporated to afford compound 4 (176 g, 97.4%). 1 H NMR (CDCl 3 , 300 MHz) δ: 7.46-7.28 (m, 5H), 6.86-6.76 (m, 3H), 5.16 (s, 2H), 3.88 (s, 3H), 2.76-2.72 (m, 2H) , 2.37-2.29 (m, 1H), 1.63-1.54 (m, 3H), 0.94 (d, J = 6.9 Hz, 6H).

化合物5的制备：化合物4(3.0g，10mmol)在乙腈(84mL)中搅拌30分钟，然后加入R-扁桃酸(0.84g，5.5mmol)并在55℃搅拌过夜。冷至室温，过滤，固体用乙腈(16mL)洗涤后干燥，得1.6g。重新加到水(8mL)中，加入1M碳酸钾水溶液(4.2mL，4.2mmol)，在室温下搅拌2小时至澄清。往体系中加入二氯甲烷(15mL)和食盐固体(0.4g)，分出有机相，饱和食盐水洗，干燥，旋干后得到化合物5(0.80g，26.6％，98％ee)。Preparation of Compound 5: Compound 4 (3.0 g, 10 mmol) was stirred in EtOAc (EtOAc) (EtOAc) It was cooled to room temperature, filtered, and the solid was washed with acetonitrile (16 mL). It was re-added to water (8 mL), and a 1M aqueous potassium carbonate solution (4.2 mL, 4.2 mmol) was added and stirred at room temperature for 2 hr. Dichloromethane (15 mL) and a salt solid (0.4 g) were added to the system, and the organic phase was separated, washed with saturated brine, dried and evaporated to give compound 5 (0.80 g, 26.6%, 98% ee).

化合物6的制备：甲酸(86.2g，1.87mol)加入到化合物5(75g，0.25mol)的甲基四氢呋喃(670mL)溶液中，回流12小时。冷却后浓缩，剩余物溶于乙酸乙酯，然后用水洗，饱和食盐水洗，无水硫酸钠干燥，旋干得到化合物6粗品(70g)，直接进行下一步反应。Preparation of Compound 6: Formic acid (86.2 g, 1.87 mol) was added to a solution of compound 5 (75 g, 0.25 mol) in methyltetrahydrofuran (670 mL) and refluxed for 12 hr. After cooling, the residue was dissolved in ethyl acetate. EtOAc (EtOAc m.

化合物7的制备：化合物6(70g，0.21mol)溶于乙腈(622mL)中，加入三氯氧磷(41g，0.27mol)，然后在90℃搅拌2小时。反应结束后冷却，在冰水浴下加入水淬灭反应，浓缩，加入1N氢氧化钠调节至pH＝10，然后用二氯 甲烷萃取(200mL×3)，有机相合并，饱和食盐水洗涤，干燥浓缩后经柱层析纯化得到化合物7(57g，86.1％)。1HNMR(CDCl3，300MHz)δ：8.39(s，1H)，7.47-7.34(m，5H)，6.92(s，1H)，6.74(s，1H)，5.22(s，2H)，3.93(s，3H)，3.42-3.39(m，1H)，2.67-2.63(m，2H)，2.11-2.09(m，1H)，1.11-1.01(m，6H)；ESI-MS m/z 310.2(M+H)+。Preparation of Compound 7: Compound 6 (70 g, 0.21 mol) was dissolved in acetonitrile (622 mL), and phosphorus oxychloride (41 g, 0.27 mol) was added, followed by stirring at 90 ° C for 2 hours. After the reaction was completed, it was cooled, and the mixture was quenched with water, and the mixture was concentrated with water, and then concentrated, and then, with 1N sodium hydroxide, adjusted to pH=10, then extracted with dichloromethane (200 mL×3), and the organic phases were combined, washed with brine and dried. After concentration, purification by column chromatography gave Compound 7 (57 g, 86.1%). 1 H NMR (CDCl 3 , 300 MHz) δ: 8.39 (s, 1H), 7.47-7.34 (m, 5H), 6.92 (s, 1H), 6.74 (s, 1H), 5.22 (s, 2H), 3.93 (s) , 3H), 3.42-3.39 (m, 1H), 2.67-2.63 (m, 2H), 2.11-2.09 (m, 1H), 1.11-1.01 (m, 6H); ESI-MS m/z 310.2 (M+ H) + .

化合物8的制备：化合物7(12.5g，40.5mmol)，2-乙氧基甲基乙酰乙酸乙酯(22.7g，0.12mol)加入到水(118mL)中，于85℃搅拌29小时。反应结束后，冷至室温，用乙酸乙酯萃取，合并有机相，饱和食盐水洗，干燥，浓缩得到24.5g粗产物。Preparation of Compound 8: Compound 7 (12.5 g, 40.5 mmol), ethyl 2-ethoxymethylacetoacetate (22.7 g, 0.12 mol) was added to water (118 mL) and stirred at 85 ° C for 29 hours. After completion of the reaction, the mixture was cooled to room temperature and extracted with ethyl acetate.

化合物9的制备：化合物8(50g，0.11mol)和四氯苯醌(27.6g，0.11mol)加入到乙二醇二甲醚(470mL)中，于70℃搅拌3小时。反应结束后过滤，滤液浓缩，然后倒入水中，二氯甲烷萃取(200mL×3)。有机相合并，饱和食盐水洗，干燥浓缩后粗产品经柱层析纯化得到化合物9(23.1g，46.2％)。1HNMR(CDCl3，300MHz)δ：8.17(s，1H)，7.46-7.34(m，5H)，7.20(s，1H)，6.92(s，1H)，6.72(s，1H)，5.22(s，2H)，4.41(q，2H)，3.94(s，3H)，3.72-2.69(m，1H)，3.29-3.23(m，1H)，2.98-2.93(m，1H)，1.80-1.72(m，1H)，1.41(m，3H)，0.89-0.78(m，6H)。Preparation of Compound 9: Compound 8 (50 g, 0.11 mol) and tetrachlorophenylhydrazine (27.6 g, 0.11 mol) were added to ethylene glycol dimethyl ether (470 mL), and stirred at 70 ° C for 3 hours. After completion of the reaction, the mixture was filtered, and the filtrate was concentrated, then poured into water, and extracted with dichloromethane (200 mL × 3). The organic phases were combined, washed with saturated brine, dried and evaporated, then evaporated 1 H NMR (CDCl 3 , 300 MHz) δ: 8.17 (s, 1H), 7.46-7.34 (m, 5H), 7.20 (s, 1H), 6.92 (s, 1H), 6.72 (s, 1H), 5.22 (s) , 2H), 4.41 (q, 2H), 3.94 (s, 3H), 3.72-2.69 (m, 1H), 3.29-3.23 (m, 1H), 2.98-2.93 (m, 1H), 1.80-1.72 (m , 1H), 1.41 (m, 3H), 0.89-0.78 (m, 6H).

化合物10的制备：Pd/C(6g)加入到化合物9(23.0g，51.4mmol)的乙醇溶液(250mL)中，反应在氢气下搅拌15小时。过滤，滤液旋干，然后用二氯甲烷重结晶得到化合物10(10.0g，54.0％)。1HNMR(CDCl3，300MHz)：δ8.28(s，1H)，7.17(s，1H)，7.07(s，1H)，6.87(s，1H)，4.41(q，J＝6.9Hz，2H)，3.94(s，3H)，3.85-3.81(m，1H)，3.33-3.26(m，1H)，3.04-2.97(m，1H)，1.84-1.76(m，1H)，1.41(t，J＝6.9Hz，3H)，0.94-0.92(m，3H)，0.84-0.82(m，3H)；ESI-MS m/z 358.1(M+H)+。Preparation of Compound 10: Pd/C (6 g) was added to a solution of compound 9 (23.0 g, 51.4 mmol) in ethanol (250 mL). Filtration, the filtrate was spun dry and then recrystallized from dichloromethane to afford compound 10 (10.0 g, 54.0%). 1 H NMR (CDCl 3 , 300 MHz): δ 8.28 (s, 1H), 7.17 (s, 1H), 7.07 (s, 1H), 6.87 (s, 1H), 4.41 (q, J = 6.9 Hz, 2H) , 3.94(s,3H), 3.85-3.81(m,1H),3.33-3.26(m,1H),3.04-2.97(m,1H),1.84-1.76(m,1H),1.41(t,J= 6.9 Hz, 3H), 0.94 - 0.92 (m, 3H), 0.84-0.82 (m, 3H); ESI-MS m/z 358.1 (M+H) + .

用同样的方法，消旋的化合物7-rac和10-rac可以用未拆分的原料制得。In the same manner, the racemic compounds 7-rac and 10-rac can be obtained from unpurified starting materials.

实施例1.制备6-异丙基-10-甲氧基-2-氧代-9-(((S)-四氢呋喃-3-基)氧基)-6，7-二氢-2H-吡啶并[2，1-α]异喹啉-3-羧酸(I-1-rac)Example 1. Preparation of 6-isopropyl-10-methoxy-2-oxo-9-(((S)-tetrahydrofuran-3-yl)oxy)-6,7-dihydro-2H-pyridine And [2,1-α]isoquinoline-3-carboxylic acid (I-1-rac)

Figure PCTCN2017094946-appb-000012
Figure PCTCN2017094946-appb-000012

化合物1b的制备：化合物1a(0.20g，2.27mmol)溶于6mL二氯甲烷，并在冰水浴中冷却，加入三乙胺(1mL，7.19mmol)，缓慢加入甲磺酰氯(0.39g，3.42mmol)，混合物于室温搅拌3小时，旋干，粗品溶于乙酸乙酯(60mL)，水洗，饱和碳酸钠洗，饱和食盐水洗，无水硫酸钠干燥，过滤，滤液旋干，得粗产品0.36g。Preparation of compound 1b: Compound 1a (0.20 g, 2.27 mmol) was dissolved in 6 mL of dichloromethane and cooled in ice-water bath, triethylamine (1 mL, 7.19 mmol) was added, and methanesulfonyl chloride (0.39 g, 3.42 mmol) was slowly added. The mixture was stirred at room temperature for 3 hours, and then dried. EtOAc EtOAcjjjjjjjjjjjjj .

化合物1c-rac的制备：化合物10-rac(100mg，0.28mmol)，化合物1b(93mg，0.56mmol)和碳酸钾(116mg，0.84mmol)加入5mL N，N-二甲基甲酰胺中，然后于90℃搅拌18小时。反应结束后倒入水中，乙酸乙酯(40mL×3)萃取，合并有机相，依次用水洗和饱和食盐水洗，无水硫酸钠干燥，过滤，滤液旋干，得到粗品0.128g。Preparation of compound 1c-rac: compound 10-rac (100 mg, 0.28 mmol), compound 1b (93 mg, 0.56 mmol) and potassium carbonate (116 mg, 0.84 mmol) in 5 mL of N,N-dimethylformamide, then Stir at 90 ° C for 18 hours. After the completion of the reaction, the mixture was poured into water, and ethyl acetate (40 mL × 3) was evaporated.

化合物I-1-rac的制备：往化合物1c-rac(0.128g，0.30mmol)的四氢呋喃(5mL)溶液中加入1N的氢氧化钠水溶液(1.8mL，1.80mmol)，于室温反应18小时。反应结束后加入1N盐酸调至pH＝1-2，二氯甲烷萃取(30mL×3)，合并有机层，无水硫酸钠干燥，过滤，滤液旋干，粗品用乙酸乙酯与甲基叔丁基醚重结晶得到化合物I-1-rac(34mg，28.3％)。1H NMR(400MHz，CDCl3)δ：8.44(s，1H)，7.19(s，1H)，7.05(s，1H)，6.70-6.67(m，1H)，5.06-5.00(m，1H)，4.06-4.00(m，3H)，3.97-3.92(m，1H)，3.91(s，3H)，3.88-3.84(m，1H)，3.36-3.31(m，1H)，3.21(s，3H)，3.08-3.06(m，1H)，3.04(s，1H)，3.03-3.02(m，1H)，2.27-2.18(m，2H)，1.86-1.77(m，1H)，0.94-0.93(d，J＝6.4Hz，3H)，0.84-0.82(d，J＝6.8Hz，3H)；ESI-MS m/z 400.2(M+H)+。Preparation of the compound I-1-rac: To a solution of the compound 1c- s (0.128 g, 0.30 mmol) in THF (5 mL). After the reaction, 1N hydrochloric acid was added to adjust to pH = 1-2, dichloromethane (30 mL × 3), and the organic layer was combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. Recrystallization of the ether gave the compound I-1-rac (34 mg, 28.3%). 1 H NMR (400MHz, CDCl 3 ) δ: 8.44 (s, 1H), 7.19 (s, 1H), 7.05 (s, 1H), 6.70-6.67 (m, 1H), 5.06-5.00 (m, 1H), 4.06-4.00 (m, 3H), 3.97-3.92 (m, 1H), 3.91 (s, 3H), 3.88-3.84 (m, 1H), 3.36-3.31 (m, 1H), 3.21 (s, 3H), 3.08-3.06(m,1H), 3.04(s,1H), 3.03-3.02(m,1H), 2.27-2.18(m,2H),1.86-1.77(m,1H),0.94-0.93(d,J = 6.4 Hz, 3H), 0.84 - 0.82 (d, J = 6.8 Hz, 3H); ESI-MS m/z 400.2 (M+H) + .

实施例2.制备(S)-6-异丙基-10-甲氧基-2-氧代-9-(((R)-四氢呋喃-3-基)氧基)-6，7-二氢-2H-吡啶并[2，1-α]异喹啉-3- 羧酸(I-2)Example 2. Preparation of (S)-6-isopropyl-10-methoxy-2-oxo-9-(((R)-tetrahydrofuran-3-yl)oxy)-6,7-dihydrogen -2H-pyrido[2,1-α]isoquinoline-3- Carboxylic acid (I-2)

化合物2b的制备：化合物2a(0.20g，2.27mmol)溶于二氯甲烷(5mL)中，加入三乙胺(1mL，7.19mmol)，缓慢加入甲磺酰氯(0.39g，3.41mmol)，混合物于室温搅拌3小时，旋干，粗品溶于乙酸乙酯(50mL)，然后依次用水，饱和碳酸钠溶液和饱和食盐水洗，无水硫酸钠干燥，过滤，滤液旋干得到化合物2b粗品(0.36g)。Preparation of compound 2b: Compound 2a (0.20 g, 2.27 mmol) was dissolved in dichloromethane (5 mL), triethylamine (1 mL, 7. The mixture was stirred at room temperature for 3 hr, EtOAc EtOAc (EtOAc)EtOAc. .

化合物2c的制备：化合物10(0.16g，0.45mmol)，化合物2b(0.149g，0.90mmol)溶于3mL N，N-二甲基甲酰胺，加入碳酸钾(0.186g，1.34mmol)，在90℃反应18小时。反应结束后倒入水中，乙酸乙酯萃取(40ml×3)，合并乙酸乙酯层，依次用水洗和饱和食盐水洗，无水硫酸钠干燥，过滤，滤液真空浓缩，得到化合物2c粗品(0.287g)。Preparation of Compound 2c: Compound 10 (0.16 g, 0.45 mmol), Compound 2b (0.149 g, 0.90 mmol) dissolved in 3 mL of N,N-dimethylformamide, and potassium carbonate (0.186 g, 1.34 mmol) The reaction was carried out at ° C for 18 hours. After the reaction was completed, the mixture was poured into EtOAc EtOAc (EtOAc m. ).

化合物I-2的制备：化合物2c(0.287g，0.67mmol)溶于5mL四氢呋喃中，加入1N氢氧化钠水溶液(4.0mL，4.0mmol)，于室温反应18小时。反应结束后加入1N盐酸调节至pH＝1-2，二氯甲烷萃取(30ml×3)，合并有机层，无水硫酸钠干燥，过滤，滤液旋干，粗品用乙酸乙酯与甲基叔丁基醚重结晶得到化合物I-2(0.113g，63.4％)。1H NMR(400MHz，CDCl3)δ：8.47(s，1H)，7.20(s，1H)，7.07(s，1H)，6.69(s，1H)，5.09-5.01(m，1H)，4.10-3.99(m，3H)，3.92(s，3H)，3.85-3.40(m，2H)，3.36(dd，J1＝16.4Hz，J2＝5.6Hz，1H)，3.11-3.04(m，1H)，2.29-2.23(m，2H)，1.87-1.78(m，1H)，0.95(d，J＝6.8Hz，3H)，0.85(d，J＝6.8Hz，3H)；ESI-MS m/z 400.2(M+H)+。Preparation of Compound I-2: Compound 2c (0.287 g, 0.67 mmol) was dissolved in 5 mL of THF. After the reaction was completed, 1N hydrochloric acid was added to adjust to pH = 1-2, dichloromethane (30 ml × 3), the organic layer was combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. The ether was recrystallized to give Compound I-2 (0.113 g, 63.4%). 1 H NMR (400MHz, CDCl 3 ) δ: 8.47 (s, 1H), 7.20 (s, 1H), 7.07 (s, 1H), 6.69 (s, 1H), 5.09-5.01 (m, 1H), 4.10- 3.99 (m, 3H), 3.92 (s, 3H), 3.85-3.40 (m, 2H), 3.36 (dd, J 1 = 16.4 Hz, J 2 = 5.6 Hz, 1H), 3.11-3.04 (m, 1H) , 2.29-2.23 (m, 2H), 1.87-1.78 (m, 1H), 0.95 (d, J = 6.8 Hz, 3H), 0.85 (d, J = 6.8 Hz, 3H); ESI-MS m/z 400.2 (M+H) + .

实施例3.制备6-异丙基-10-甲氧基-2-氧代-9-(((R)-四氢呋喃-3-基)氧基)-6，7-二氢-2H-吡啶并[2，1-α]异喹啉-3-羧酸(I-2-rac)Example 3. Preparation of 6-isopropyl-10-methoxy-2-oxo-9-(((R)-tetrahydrofuran-3-yl)oxy)-6,7-dihydro-2H-pyridine And [2,1-α]isoquinoline-3-carboxylic acid (I-2-rac)

Figure PCTCN2017094946-appb-000014
Figure PCTCN2017094946-appb-000014

按照实施例2的方法，得到I-2-rac，1H NMR(400MHz，CDCl3)δ：8.47(s，1H)，7.20(s，1H)，7.07(s，1H)，6.69(s，1H)，5.09-5.01(m，1H)，4.10-3.99(m，3H)，3.92(s，3H)，3.85-3.40(m，2H)，3.36(dd，J1＝16.4Hz，J2＝5.6Hz，1H)，3.11-3.04(m，1H)，2.29-2.23(m，2H)，1.87-1.78(m，1H)，0.95(d，J＝6.8Hz，3H)，0.85(d，J＝6.8Hz，3H)；ESI-MS m/z 400.2(M+H)+。I-2-rac, 1 H NMR (400 MHz, CDCl 3 ) δ: 8.47 (s, 1H), 7.20 (s, 1H), 7.07 (s, 1H), 6.69 (s, 1H), 5.09-5.01 (m, 1H), 4.10-3.99 (m, 3H), 3.92 (s, 3H), 3.85-3.40 (m, 2H), 3.36 (dd, J 1 = 16.4 Hz, J 2 = 5.6 Hz, 1H), 3.11-3.04 (m, 1H), 2.29-2.23 (m, 2H), 1.87-1.78 (m, 1H), 0.95 (d, J = 6.8 Hz, 3H), 0.85 (d, J) = 6.8 Hz, 3H); ESI-MS m/z 400.2 (M+H) + .

实施例4.制备6-异丙基-10-甲氧基-2-氧代-9-((四氢-2H-吡喃-4-基)氧基)-6，7-二氢-2H-吡啶并[2，1-α]异喹啉-3-羧酸(I-3-rac)Example 4. Preparation of 6-isopropyl-10-methoxy-2-oxo-9-((tetrahydro-2H-pyran-4-yl)oxy)-6,7-dihydro-2H -pyrido[2,1-α]isoquinoline-3-carboxylic acid (I-3-rac)

Figure PCTCN2017094946-appb-000015
Figure PCTCN2017094946-appb-000015

化合物3a-rac的制备：化合物10-rac(100mg，0.28mmol)和4-溴四氢吡喃(94mg，0.56mmol)溶于5mL N，N二甲基甲酰胺，加入碳酸钾(78mg，0.56mmol)，然后在90℃反应40小时。反应结束后倒入水中，乙酸乙酯萃取(40mL×3)，合并乙酸乙酯层，依次用水洗和饱和食盐水洗，无水硫酸钠干燥，过滤，滤液真空浓缩，粗产品经制备板纯化得化合物3a-rac(65mg，56.1％)。 Preparation of compound 3a-rac: Compound 10-rac (100 mg, 0.28 mmol) and 4-bromotetrahydropyran (94 mg, 0.56 mmol) were dissolved in 5 mL of N, N dimethylformamide and potassium carbonate (78 mg, 0.56) Methyl), then reacted at 90 ° C for 40 hours. After the reaction was completed, the mixture was poured into water and ethyl acetate (40 mL×3). Compound 3a-rac (65 mg, 56.1%).

化合物I-3-rac的制备：化合物3a-rac(65mg，0.15mmol)溶于四氢呋喃(5mL)中，加入1N氢氧化钠水溶液(1.0mL，1.0mmol)，于室温反应18小时。反应结束后加入1N盐酸调节至pH＝1-2，二氯甲烷萃取(30ml×3)，合并有机层，无水硫酸钠干燥，过滤，滤液旋干，粗品经制备板纯化得化合物I-3-rac(2mg，3.2％)。1H NMR(400MHz，CDCl3)δ：15.94(s，1H)，8.44(s，1H)，7.19(s，1H)，7.06(s，1H)，6.77(s，1H)，4.57(m，2H)，4.03-4.02(m，2H)，3.92(s，3H)，3.87-3.84(m，1H)，3.61-3.55(m，2H)，3.35-3.30(m，1H)，3.07-3.03(m，1H)，2.07-2.04(m，1H)，1.81-1.93(m，4H)，0.94-0.93(d，J＝6.4Hz，3H)，0.84-0.82(d，J＝6.8Hz，3H)；ESI-MS m/z 414.2(M+H)+。Preparation of Compound I-3-rac: Compound 3a-rac (65 mg, 0.15 mmol) was dissolved in THF (5 mL). After the completion of the reaction, 1N hydrochloric acid was added to adjust to pH = 1-2, dichloromethane (30 ml × 3), the organic layer was combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. -rac (2mg, 3.2%). 1 H NMR (400MHz, CDCl 3 ) δ: 15.94 (s, 1H), 8.44 (s, 1H), 7.19 (s, 1H), 7.06 (s, 1H), 6.77 (s, 1H), 4.57 (m, 2H), 4.03-4.02 (m, 2H), 3.92 (s, 3H), 3.87-3.84 (m, 1H), 3.61-3.55 (m, 2H), 3.35-3.30 (m, 1H), 3.07-3.03 ( m, 1H), 2.07-2.04 (m, 1H), 1.81-1.93 (m, 4H), 0.94-0.93 (d, J = 6.4 Hz, 3H), 0.84-0.82 (d, J = 6.8 Hz, 3H) ;ESI-MS m/z 414.2 (M+H) + .

实施例5.制备6-异丙基-10-甲氧基-9-(氧杂环丁烷-3-基氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-4-rac)Example 5. Preparation of 6-isopropyl-10-methoxy-9-(oxetan-3-yloxy)-2-oxo-6,7-dihydro-2H-pyridine and [ 2,1-a]isoquinoline-3-carboxylic acid (I-4-rac)

Figure PCTCN2017094946-appb-000016
Figure PCTCN2017094946-appb-000016

化合物4a-rac的制备：化合物10-rac(100mg，0.28mmol)和3-溴氧杂环丁烷(75mg，0.56mmol)溶于N，N-二甲基甲酰胺(5mL)，加入碳酸钾(78mg，0.56mmol)，在90℃搅拌40小时。反应结束后倒入水中，乙酸乙酯萃取(40mL×3)，合并乙酸乙酯层，依次用水洗和饱和食盐水洗，无水硫酸钠干燥，过滤，滤液旋干，粗品经制备板分离得到化合物4a-rac(51mg，42.9％)。Preparation of compound 4a-rac: Compound 10-rac (100 mg, 0.28 mmol) and 3-bromo oxetane (75 mg, 0.56 mmol) dissolved in N,N-dimethylformamide (5 mL) (78 mg, 0.56 mmol), stirred at 90 ° C for 40 hours. After the reaction was completed, the mixture was poured into water, ethyl acetate (40 mL × 3), and the ethyl acetate layer was combined, washed successively with water and brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. 4a-rac (51 mg, 42.9%).

化合物I-4-rac的制备：化合物4a-rac(50mg，0.12mmol)溶于四氢呋喃(5mL)，加入1N氢氧化钠水溶液(1.0mL，1.0mmol)，于室温反应18小时。反应结束后加入1N盐酸调节至pH＝1-2，二氯甲烷萃取(30ml×3)，合并有机层，无水硫酸钠干燥，过滤，滤液旋干，粗品经制备板纯化得化合物I-4-rac(7mg，15.1％)。1H NMR(400MHz，CDCl3)δ：15.87(s，1H)，8.45(s，1H)，7.21(s，1H)，7.06(s 1H)，6.33(s，1H)，4.98(s，2H)，4.82(s，2H)，3.94(s，5H)，3.30(s，1H)，3.03(s，1H)，1.78(s，1H)，0.92(s，3H)，0.82(s，3H)；ESI-MS m/z 386.2(M+H)+。Preparation of Compound I-4-rac: Compound 4a-rac (50 mg, 0.12 mmol) was dissolved in THF (5 mL). After the completion of the reaction, 1N hydrochloric acid was added to adjust to pH = 1-2, dichloromethane (30 ml × 3), the organic layer was combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. -rac (7mg, 15.1%). 1 H NMR (400MHz, CDCl 3 ) δ: 15.87 (s, 1H), 8.45 (s, 1H), 7.21 (s, 1H), 7.06 (s 1H), 6.33 (s, 1H), 4.98 (s, 2H ), 4.82 (s, 2H), 3.94 (s, 5H), 3.30 (s, 1H), 3.03 (s, 1H), 1.78 (s, 1H), 0.92 (s, 3H), 0.82 (s, 3H) ;ESI-MS m/z 386.2 (M+H) + .

实施例6.制备(S)-6-异丙基-10-甲氧基-9-((1R，3S)-3-甲氧基环丁氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-α]异喹啉-3-羧酸(I-5)Example 6. Preparation of (S)-6-isopropyl-10-methoxy-9-((1R,3S)-3-methoxycyclobutoxy)-2-oxo-6,7- Dihydro-2H-pyrido[2,1-α]isoquinoline-3-carboxylic acid (I-5)

Figure PCTCN2017094946-appb-000017
Figure PCTCN2017094946-appb-000017

化合物5b的制备：化合物5a(0.70g，3.64mmol)溶于100mL甲醇，加入Pd/C(0.12g)和1滴浓盐酸，氢气置换3次，室温加氢18小时，过滤，滤液旋干得到化合物5b粗品0.43g。Preparation of compound 5b: Compound 5a (0.70 g, 3.64 mmol) was dissolved in 100 mL of methanol, Pd/C (0.12 g) and 1 drop of concentrated hydrochloric acid were added, and hydrogen was replaced three times, hydrogenated at room temperature for 18 hours, filtered, and the filtrate was dried. The crude compound 5b was 0.43 g.

化合物5c的制备：化合物5b(0.43g，4.22mmol)溶于8mL二氯甲烷中，加入三乙胺(1.5mL，8.63mmol)，缓慢加入甲磺酰氯(0.72g，6.32mmol)，混合物于室温搅拌3小时，旋干，粗品溶于乙酸乙酯(50mL)，然后依次用水，饱和碳酸钠溶液和饱和食盐水洗，无水硫酸钠干燥，过滤，滤液旋干得到化合物5c粗品0.70g。Preparation of compound 5c: Compound 5b (0.43 g, 4.22 mmol) was dissolved in dichloromethane (3 mL), triethylamine (1.5 mL, 8.63 mmol), methanesulfonyl chloride (0.72 g, 6.32 mmol) After stirring for 3 hours, the mixture was evaporated to dryness crystals crystals crystals crystals crystals

化合物5d的制备：化合物10(0.160g，0.45mmol)和化合物5c(0.161g，0.90mmol)溶于N，N-二甲基甲酰胺(5mL)，加入碳酸钾(0.185g，1.34mmol)，在90℃搅拌18小时，倒入水中，乙酸乙酯萃取(30mL×4)，合并乙酸乙酯层，水洗，饱和食盐水洗，无水硫酸钠干燥，过滤，滤液旋干得到油状物0.21g，为化合物5d。 Preparation of compound 5d: Compound 10 (0.160 g, 0.45 mmol) and compound 5c (0.161 g, 0.90 mmol) were dissolved in N,N-dimethylformamide (5 mL), and potassium carbonate (0.185 g, 1. The mixture was stirred at 90 ° C for 18 hours, poured into water, ethyl acetate (30 mL × 4), ethyl acetate layer, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and evaporated. Is compound 5d.

化合物I-5的制备：化合物5d(0.210g，0.48mmol)溶于四氢呋喃(5mL)，加入1N氢氧化钠水溶液(2.85mL，2.85mmol)，室温反应18小时。反应结束后加入1N盐酸调节至pH＝1-2，二氯甲烷萃取(30ml×3)，合并有机层，无水硫酸钠干燥，过滤，滤液旋干，粗品经制备板纯化得化合物I-5(64mg，32.2％)。1H NMR(400MHz，CDCl3)δ：8.46(s，1H)，7.18(s，1H)，7.07(s，1H)，6.54(s，1H)，5.00-4.90(m，1H)，4.20-4.13(m，1H)，3.94(s，3H)，3.91-3.84(m，1H)，3.52-3.34(m，1H)，3.31(s，3H)，2.59-2.46(m，1H)，2.59-2.48(m，4H)，1.88-1.78(m，1H)，0.95(d，J＝6.8Hz，3H)，0.84(d，J＝6.8Hz，3H)；ESI-MS m/z 414.2(M+H)+。Preparation of Compound I-5: Compound 5d (0.210 g, 0.48 mmol) was dissolved in THF (5 mL). After the completion of the reaction, 1N hydrochloric acid was added to adjust to pH = 1-2, dichloromethane (30 ml × 3), the organic layer was combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. (64 mg, 32.2%). 1 H NMR (400MHz, CDCl 3 ) δ: 8.46 (s, 1H), 7.18 (s, 1H), 7.07 (s, 1H), 6.54 (s, 1H), 5.00-4.90 (m, 1H), 4.20- 4.13 (m, 1H), 3.94 (s, 3H), 3.91-3.84 (m, 1H), 3.52-3.34 (m, 1H), 3.31 (s, 3H), 2.59-2.46 (m, 1H), 2.59- 2.48 (m, 4H), 1.88-1.78 (m, 1H), 0.95 (d, J = 6.8 Hz, 3H), 0.84 (d, J = 6.8 Hz, 3H); ESI-MS m/z 414.2 (M+ H) + .

实施例7.制备6-异丙基-10-甲氧基-9-((1R，3S)-3-甲氧基环丁氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-α]异喹啉-3-羧酸(I-5-rac)Example 7. Preparation of 6-isopropyl-10-methoxy-9-((1R,3S)-3-methoxycyclobutoxy)-2-oxo-6,7-dihydro-2H -pyrido[2,1-α]isoquinoline-3-carboxylic acid (I-5-rac)

Figure PCTCN2017094946-appb-000018
Figure PCTCN2017094946-appb-000018

按照实施例6的方法，得到化合物I-5-rac，1H NMR(400MHz，CDCl3)δ：8.46(s，1H)，7.18(s，1H)，7.07(s，1H)，6.54(s，1H)，5.00-4.90(m，1H)，4.20-4.13(m，1H)，3.94(s，3H)，3.91-3.84(m，1H)，3.52-3.34(m，1H)，3.31(s，3H)，2.59-2.46(m，1H)，2.59-2.48(m，4H)，1.88-1.78(m，1H)，0.95(d，J＝6.8Hz，3H)，0.84(d，J＝6.8Hz，3H)；ESI-MS m/z414.2(M+H)+。The compound I-5-rac, 1 H NMR (400 MHz, CDCl 3 ) δ: 8.46 (s, 1H), 7.18 (s, 1H), 7.07 (s, 1H), 6.54 (s) , 1H), 5.00-4.90 (m, 1H), 4.20-4.13 (m, 1H), 3.94 (s, 3H), 3.91-3.84 (m, 1H), 3.52-3.34 (m, 1H), 3.31 (s , 3H), 2.59-2.46 (m, 1H), 2.59-2.48 (m, 4H), 1.88-1.78 (m, 1H), 0.95 (d, J = 6.8 Hz, 3H), 0.84 (d, J = 6.8 Hz, 3H); ESI-MS m/z 414.2 (M+H) + .

实施例8制备(S)-6-异丙基-10-甲氧基-9-((1-(甲氧基甲基)环丙基)甲氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-α]异喹啉-3-羧酸(I-6)Example 8 Preparation of (S)-6-isopropyl-10-methoxy-9-((1-(methoxymethyl)cyclopropyl)methoxy)-2-oxo-6,7 -Dihydro-2H-pyrido[2,1-α]isoquinoline-3-carboxylic acid (I-6)

Figure PCTCN2017094946-appb-000019
Figure PCTCN2017094946-appb-000019

化合物6b的制备：化合物6a(0.50g，4.9mmol)，碳酸钾(1.02g，7.4mmol)，碘甲烷(1.04g，7.4mmol)和4-氟苯硼酸(69mg，0.49mmol)在N，N-二甲基甲酰胺(5mL)中室温搅拌16小时。往反应液中加入30mL水，然后用乙酸乙酯萃取(30ml×3)。有机相合并，用饱和食盐水洗一次，无水硫酸钠干燥，旋干后得到200mg化合物6b的粗品。直接用于下一步反应。1H NMR(400MHz，CDCl3)δ：3.54(d，J＝5.2Hz，2H)，3.38(s，2H)，3.37(s，3H)，2.51(t，J＝5.2Hz，1H)，0.50-0.55(m，4H)。Preparation of compound 6b: Compound 6a (0.50 g, 4.9 mmol), potassium carbonate (1.02 g, 7.4 mmol), iodomethane (1.04 g, 7.4 mmol) and 4-fluorophenylboronic acid (69 mg, 0.49 mmol) in N, N -Methylformamide (5 mL) was stirred at room temperature for 16 hours. 30 mL of water was added to the reaction mixture, followed by extraction with ethyl acetate (30 ml × 3). The organic phases were combined, washed once with brine, dried over anhydrous sodium sulfate and evaporated Used directly in the next step. 1 H NMR (400MHz, CDCl 3 ) δ: 3.54 (d, J = 5.2Hz, 2H), 3.38 (s, 2H), 3.37 (s, 3H), 2.51 (t, J = 5.2Hz, 1H), 0.50 -0.55 (m, 4H).

化合物6c的制备：化合物6b(0.20g，1.72mmol)溶于10mL二氯甲烷并用冰水浴冷却。然后依次加入三乙胺(0.35 g，3.44mmol)和甲基磺酰氯(0.30g，2.59mmol)。在室温下反应2小时后，加入二氯甲烷(20mL)稀释，并用1N稀盐酸(10mL)洗涤。水相用二氯甲烷(20mL×2)萃取。合并有机相，饱和食盐水洗涤，无水硫酸钠干燥之后旋干得到0.30g黄色油状物，为化合物6c。Preparation of compound 6c: Compound 6b (0.20 g, 1.72 mmol) was dissolved in 10 mL dichloromethane and cooled with ice water. Then add triethylamine (0.35) g, 3.44 mmol) and methanesulfonyl chloride (0.30 g, 2.59 mmol). After reacting for 2 hours at room temperature, it was diluted with dichloromethane (20 mL) and washed with 1 N diluted hydrochloric acid (10 mL). The aqueous phase was extracted with dichloromethane (20 mL x 2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate

化合物6d的制备：化合物10(0.15g，0.42mmol)，化合物6c(0.30g，1.54mmol)和无水碳酸钾(0.12g，0.84mmol)在5mL N，N-二甲基甲酰胺中于90℃搅拌16小时。反应结束后冷至室温，加入30mL水稀释，并用乙酸乙酯萃取(20mL×2)。合并有机相，饱和食盐水洗，无水硫酸钠干燥后浓缩得到0.18g化合物6d的粗品，直接用于下一步反应。Preparation of compound 6d: compound 10 (0.15 g, 0.42 mmol), compound 6c (0.30 g, 1.54 mmol) and anhydrous potassium carbonate (0.12 g, 0.84 mmol) in 5 mL of N,N-dimethylformamide Stir at °C for 16 hours. After the reaction was completed, it was cooled to room temperature, diluted with water (30 mL), and extracted with ethyl acetate (20 mL×2). The organic phase was combined, washed with brine, dried over anhydrous sodium sulfate and evaporated.

化合物I-6的制备：往化合物6d(0.18g，0.40mmol)的甲醇(10mL)溶液中加入1N氢氧化钠水溶液(1.6mL，1.60mmol)，于50℃反应1.5小时。反应结束后加入1N盐酸调节至pH＝1-2，二氯甲烷萃取(20mL×3)，合并有机层，无水硫酸钠干燥，过滤，滤液旋干，粗品经制备板纯化得产物化合物I-6(26mg，15.2％)。1H NMR(400MHz，CDCl3)δ：8.47(s，1H)，7.17(s，1H)，7.07(s，1H)，6.76(s，1H)，3.98(m，2H)，3.91(s，3H)，3.88(m，1H)，3.40(m，2H)，3.35(s，3H)，3.29-3.34(m，1H)，3.02-3.06(m，1H)，1.82(m，1H)，0.92(d，J＝6.8Hz，3H)，0.81(d，J＝6.8Hz，3H)，0.67(m，2H)，0.64(m，2H)；ESI-MS m/z 428.2(M+H)+。Preparation of Compound I-6: To a solution of compound 6d (0.18 g, 0.40 mmol) in methanol (10 mL) After the reaction, 1N hydrochloric acid was added to adjust to pH = 1-2, dichloromethane (20 mL × 3), and the organic layer was combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. 6 (26 mg, 15.2%). 1 H NMR (400MHz, CDCl 3 ) δ: 8.47 (s, 1H), 7.17 (s, 1H), 7.07 (s, 1H), 6.76 (s, 1H), 3.98 (m, 2H), 3.91 (s, 3H), 3.88 (m, 1H), 3.40 (m, 2H), 3.35 (s, 3H), 3.29-3.34 (m, 1H), 3.02-3.06 (m, 1H), 1.82 (m, 1H), 0.92 (d, J = 6.8 Hz, 3H), 0.81 (d, J = 6.8 Hz, 3H), 0.67 (m, 2H), 0.64 (m, 2H); ESI-MS m/z 428.2 (M+H) + .

实施例9.制备6-异丙基-10-甲氧基-9-((1-(甲氧基甲基)环丙基)甲氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-α]异喹啉-3-羧酸(I-6-rac)Example 9. Preparation of 6-isopropyl-10-methoxy-9-((1-(methoxymethyl)cyclopropyl)methoxy)-2-oxo-6,7-dihydro -2H-pyrido[2,1-α]isoquinoline-3-carboxylic acid (I-6-rac)

Figure PCTCN2017094946-appb-000020
Figure PCTCN2017094946-appb-000020

按照实施例8的方法，得到化合物I-6-rac，1H NMR(400MHz，CDCl3)δ：8.47(s，1H)，7.17(s，1H)，7.07(s，1H)，6.76(s，1H)，3.98(m，2H)，3.91(s，3H)，3.88(m，1H)，3.40(m，2H)，3.35(s，3H)，3.29-3.34(m，1H)，3.02-3.06(m，1H)，1.82(m，1H)，0.92(d，J＝6.8Hz，3H)，0.81(d，J＝6.8Hz，3H)，0.67(m，2H)，0.64(m，2H)；ESI-MS m/z 428.2(M+H)+。The compound I-6-rac, 1 H NMR (400 MHz, CDCl 3 ) δ: 8.47 (s, 1H), 7.17 (s, 1H), 7.07 (s, 1H), 6.76 (s) , 1H), 3.98 (m, 2H), 3.91 (s, 3H), 3.88 (m, 1H), 3.40 (m, 2H), 3.35 (s, 3H), 3.29-3.34 (m, 1H), 3.02- 3.06 (m, 1H), 1.82 (m, 1H), 0.92 (d, J = 6.8 Hz, 3H), 0.81 (d, J = 6.8 Hz, 3H), 0.67 (m, 2H), 0.64 (m, 2H) ESI-MS m/z 428.2 (M+H) + .

实施例10.制备(S)-6-异丙基-10-甲氧基-9-((3-(甲氧基甲基)氧杂环丁烷-3-基)甲氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-α]异喹啉-3-羧酸(I-7)Example 10. Preparation of (S)-6-isopropyl-10-methoxy-9-((3-(methoxymethyl)oxetan-3-yl)methoxy)-2 -oxo-6,7-dihydro-2H-pyrido[2,1-α]isoquinoline-3-carboxylic acid (I-7)

Figure PCTCN2017094946-appb-000021
Figure PCTCN2017094946-appb-000021

化合物7b的制备：化合物7a(0.27g，2.3mmol)，无水碳酸钾(0.48g，3.5mmol)，苄溴(0.59g，3.5mmol)和 4-氟苯硼酸(32mg，0.23mmol)在2mL N，N-二甲基甲酰胺中室温搅拌两天。往反应液中加入20mL水，然后用乙酸乙酯萃取(20mL×3)。有机相合并，用饱和食盐水洗，然后用无水硫酸钠干燥后旋干。粗品用柱层析分离得到0.28g油状产物，即为化合物7b。1H NMR(400MHz，CDCl3)δ：7.31-7.36(m，5H)，4.56(s，2H)，4.48(d，J＝6.4Hz，2H)，4.42(d，J＝6.0Hz，2H)，3.93(d，J＝4.4Hz，2H)，3.80(s，2H)，2.34(t，J＝5.2Hz，1H)。Preparation of compound 7b: Compound 7a (0.27 g, 2.3 mmol), anhydrous potassium carbonate (0.48 g, 3.5 mmol), benzyl bromide (0.59 g, 3.5 mmol) and 4-fluorophenylboronic acid (32 mg, 0.23 mmol) in 2 mL N,N-dimethylformamide was stirred at room temperature for two days. 20 mL of water was added to the reaction mixture, followed by extraction with ethyl acetate (20 mL × 3). The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate and dried. The crude product was isolated by column chromatography to give 0.28 g of oily product. 1 H NMR (400MHz, CDCl 3 ) δ: 7.31-7.36 (m, 5H), 4.56 (s, 2H), 4.48 (d, J = 6.4Hz, 2H), 4.42 (d, J = 6.0Hz, 2H) , 3.93 (d, J = 4.4 Hz, 2H), 3.80 (s, 2H), 2.34 (t, J = 5.2 Hz, 1H).

化合物7c的制备：化合物7b(0.28g，1.35mmol)溶于干燥的N，N-二甲基甲酰胺(5mL)并用冰水浴冷却。往此溶液中加入NaH(65mg，2.7mmol)并在冰水浴中搅拌40分钟。然后加入碘甲烷(0.48g，3.38mmol)并在冰水浴中搅拌5小时。往反应液中加入30mL水淬灭，然后用乙酸乙酯萃取(20mL×3)。合并有机相，饱和食盐水洗，无水硫酸钠干燥之后旋干。粗品用柱层析分离得到无色油状物化合物7c(0.18g，60.1％)。1H NMR(400MHz，CDCl3)δ：7.29-7.36(m，5H)，4.56(s，2H)，4.48(m，4H)，3.67(s，2H)，3.62(s，2H)，3.38(s，3H)。Preparation of compound 7c: Compound 7b (0.28 g, 1.35 mmol) was dissolved in dry <RTI ID=0.0> To this solution was added NaH (65 mg, 2.7 mmol) and stirred for 40 min. Methyl iodide (0.48 g, 3.38 mmol) was then added and stirred in an ice water bath for 5 h. The reaction solution was quenched by the addition of 30 mL of water, and then ethyl acetate (20mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate and evaporated. The crude product was purified by column chromatography eluting elut elut eluting 1 H NMR (400MHz, CDCl 3 ) δ: 7.29-7.36 (m, 5H), 4.56 (s, 2H), 4.48 (m, 4H), 3.67 (s, 2H), 3.62 (s, 2H), 3.38 ( s, 3H).

化合物7d的制备：化合物7c(0.18g，0.81mmol)溶于10mL甲醇，加入Pd/C(18mg)。反应在氢气气氛中反应16小时。然后加入1滴浓盐酸，继续在氢气气氛中反应4小时。过滤，滤液旋干，并用柱层析分离得到无色液体，即为化合物7d(0.11g，100％)。1H NMR(400MHz，CDCl3)δ：4.49(d，J＝6.4Hz，2H)，4.43(d，J＝6.0Hz，2H)，3.94(m，2H)，3.74(s，2H)，3.40(s，3H)。Preparation of compound 7d: Compound 7c (0.18 g, 0.81 mmol) was dissolved in 10 mL methanol and Pd/C (18 mg). The reaction was allowed to react in a hydrogen atmosphere for 16 hours. Then, 1 drop of concentrated hydrochloric acid was added, and the reaction was continued for 4 hours in a hydrogen atmosphere. Filtration, the filtrate was spun dry, and separated by column chromatography to give a colorless liquid, Compound 7d (0.11 g, 100%). 1 H NMR (400MHz, CDCl 3 ) δ: 4.49 (d, J = 6.4Hz, 2H), 4.43 (d, J = 6.0Hz, 2H), 3.94 (m, 2H), 3.74 (s, 2H), 3.40 (s, 3H).

化合物7e的制备：化合物7d(0.11g，0.83mmol)溶于10mL二氯甲烷并用冰水浴冷却。然后依次加入三乙胺(0.168g，1.66mmol)和甲基磺酰氯(0.143g，1.25mmol)。在室温下反应2小时后，加入10mL二氯甲烷稀释，并用1N稀盐酸洗涤。水相用二氯甲烷萃取(20mL×3)。合并有机相，饱和食盐水洗涤，无水硫酸钠干燥之后旋干得到0.13g黄色油状物，即为化合物7e。Preparation of compound 7e: Compound 7d (0.11 g, 0.83 mmol) was dissolved in 10 mL dichloromethane and cooled with ice water. Then triethylamine (0.168 g, 1.66 mmol) and methanesulfonyl chloride (0.143 g, 1.25 mmol) were added sequentially. After reacting for 2 hours at room temperature, it was diluted with 10 mL of dichloromethane and washed with 1N diluted hydrochloric acid. The aqueous phase was extracted with dichloromethane (20 mL x 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate

化合物7f的制备：化合物10(0.15g，0.42mmol)，化合物7e(0.13g，0.63mmol)和无水碳酸钾(0.12g，0.84mmol)在5mL DMF中于90℃搅拌16小时。反应结束后冷至室温，加入30mL水稀释，并用乙酸乙酯萃取(20mL×3)。合并有机相，饱和食盐水洗，无水硫酸钠干燥后浓缩。粗产品经薄层层析分离后得到白色固体，为化合物7f(0.16g，76.2％)。Preparation of compound 7f: Compound 10 (0.15 g, 0.42 mmol), Compound 7e (0.13 g, 0.63 mmol) and anhydrous potassium carbonate (0.12 g, 0.84 mmol) were stirred in 5 mL DMF at 90 ° C for 16 hours. After the reaction was completed, it was cooled to room temperature, diluted with water (30 mL) and extracted with ethyl acetate (20 mL×3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate and evaporated. The crude product was purified by EtOAc (EtOAc:EtOAc)

化合物I-7的制备：往化合物7f(0.16g，0.32mmol)的甲醇(10mL)溶液中加入1N氢氧化钠水溶液(1.3mL，1.30mmol)，于50℃反应2小时。反应结束后加入1N盐酸调节至pH＝1-2，二氯甲烷萃取(20mL×3)，合并有机层，无水硫酸钠干燥，过滤，滤液旋干得到白色固体化合物I-7(94mg，66.2％)。1H NMR(400MHz，CDCl3)δ：8.45(s，1H)，7.18(s，1H)，7.06(s，1H)，6.82(s，1H)，4.56-4.63(m，4H)，4.28(m，2H)，3.90(s，3H)，3.86(m，1H)，3.77(m，2H)，3.40(s，3H)，3.32-3.36(m，1H)，3.05-3.09(m，1H)，1.84(m，1H)，0.95(d，J＝6.8Hz，3H)，0.83(d，J＝6.8Hz，3H)；ESI-MS m/z 444.2(M+H)+。Preparation of Compound I-7: To a solution of Compound 7f (0.16 g, 0.32 mmol) in methanol (10 mL). After the reaction was completed, 1N hydrochloric acid was added to adjust to pH = 1-2, dichloromethane (20 mL × 3), and the organic layer was combined, dried over anhydrous sodium sulfate, filtered and evaporated to give a white solid compound I-7 (94 mg, 66.2 %). 1 H NMR (400MHz, CDCl 3 ) δ: 8.45 (s, 1H), 7.18 (s, 1H), 7.06 (s, 1H), 6.82 (s, 1H), 4.56-4.63 (m, 4H), 4.28 ( m, 2H), 3.90 (s, 3H), 3.86 (m, 1H), 3.77 (m, 2H), 3.40 (s, 3H), 3.32-3.36 (m, 1H), 3.05-3.09 (m, 1H) , 1.84 (m, 1H), 0.95 (d, J = 6.8 Hz, 3H), 0.83 (d, J = 6.8 Hz, 3H); ESI-MS m/z 444.2 (M+H) + .

实施例11.制备6-异丙基-10-甲氧基-9-((3-(甲氧基甲基)氧杂环丁烷-3-基)甲氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-α]异喹啉-3-羧酸(I-7-rac)Example 11. Preparation of 6-isopropyl-10-methoxy-9-((3-(methoxymethyl)oxetan-3-yl)methoxy)-2-oxo- 6,7-Dihydro-2H-pyrido[2,1-α]isoquinoline-3-carboxylic acid (I-7-rac)

Figure PCTCN2017094946-appb-000022
Figure PCTCN2017094946-appb-000022

按照实施例10的方法，得到I-7-rac。1H NMR(400MHz，CDCl3)δ：8.45(s，1H)，7.18(s，1H)，7.06(s，1H)，6.82(s，1H)，4.56-4.63(m，4H)，4.28(m，2H)，3.90(s，3H)，3.86(m，1H)，3.77(m，2H)，3.40(s，3H)，3.32-3.36(m，1H)，3.05-3.09(m，1H)，1.84(m，1H)，0.95(d，J＝6.8Hz，3H)，0.83(d，J＝6.8Hz，3H)；ESI-MS m/z 444.2(M+H)+。 According to the method of Example 10, I-7-rac was obtained. 1 H NMR (400MHz, CDCl 3 ) δ: 8.45 (s, 1H), 7.18 (s, 1H), 7.06 (s, 1H), 6.82 (s, 1H), 4.56-4.63 (m, 4H), 4.28 ( m, 2H), 3.90 (s, 3H), 3.86 (m, 1H), 3.77 (m, 2H), 3.40 (s, 3H), 3.32-3.36 (m, 1H), 3.05-3.09 (m, 1H) , 1.84 (m, 1H), 0.95 (d, J = 6.8 Hz, 3H), 0.83 (d, J = 6.8 Hz, 3H); ESI-MS m/z 444.2 (M+H) + .

实施例12.制备(S)-6-异丙基-10-甲氧基-9-((3-甲氧基环丁基)甲氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-8)Example 12. Preparation of (S)-6-isopropyl-10-methoxy-9-((3-methoxycyclobutyl)methoxy)-2-oxo-6,7-dihydrogen -2H-pyrido[2,1-a]isoquinoline-3-carboxylic acid (I-8)

Figure PCTCN2017094946-appb-000023
Figure PCTCN2017094946-appb-000023

化合物8b的制备：将化合物8a(5.0g，38mmol)溶于50mL四氢呋喃中，冰浴条件下分批次加入氢化钠(1.84g，76mmol)，加毕继续搅拌反应30分钟，缓慢滴加碘甲烷(2.87mL，46mmol)。滴毕室温搅拌反应3小时，TLC监测原料反应完毕。将反应液倒入饱和氯化铵溶液中淬灭，乙酸乙酯萃取(30mL×3)。合并有机相，无水硫酸钠干燥，硅胶柱层析(乙酸乙酯∶石油醚＝1∶5)得化合物8b(1.90g，34.2％)。1H NMR(400MHz，CDCl3)δ：3.81-3.73(m，1H)，3.66(s，3H)，3.21(s，3H)，2.66-2.58(m，1H)，2.51-2.45(m，2H)，2.21-2.13(m，2H)。Preparation of compound 8b: Compound 8a (5.0 g, 38 mmol) was dissolved in 50 mL of tetrahydrofuran, and sodium hydride (1.84 g, 76 mmol) was added in portions under ice-cooling, and stirring was continued for 30 minutes, and iodomethane was slowly added dropwise. (2.87 mL, 46 mmol). The reaction was stirred at room temperature for 3 hours, and the reaction of the starting material was monitored by TLC. The reaction solution was poured into a saturated aqueous solution of ammonium chloride and evaporated, and ethyl acetate (30mL, 3). The organic phase was combined, dried over anhydrous sodium sulfate EtOAcjjjjjj 1 H NMR (400MHz, CDCl 3 ) δ: 3.81-3.73 (m, 1H), 3.66 (s, 3H), 3.21 (s, 3H), 2.66-2.58 (m, 1H), 2.51-2.45 (m, 2H ), 2.21-2.13 (m, 2H).

化合物8c的制备：将化合物8b(1.90g，13mmol)溶于20mL无水四氢呋喃中，冰浴条件下，氮气置换气3次，缓慢滴加1.5M DIBAL-H的四氢呋喃溶液(26.4mL，39mmol)。滴毕继续搅拌反应过夜，TLC监测反应完毕，加入2N盐酸(20mL)淬灭反应，乙酸乙酯(30mL×3)萃取，合并有机相，饱和食盐水洗涤，无水硫酸钠干燥，浓缩蒸干硅胶柱层析(乙酸乙酯∶石油醚＝1∶3)得化合物8c(1.00g，66.2％)。1H NMR(400MHz，CDCl3)δ：3.80-3.73(m，1H)，3.62-3.57(m，2H)，3.21(s，3H)，2.37-2.30(m，2H)，2.08-2.02(m，1H)，1.67-1.60(m，2H)。Preparation of compound 8c: Compound 8b (1.90 g, 13 mmol) was dissolved in 20 mL of anhydrous tetrahydrofuran, and the gas was replaced with nitrogen three times in ice bath, and a solution of 1.5 M DIBAL-H in tetrahydrofuran (26.4 mL, 39 mmol) was slowly added dropwise. . After the completion of the dropwise addition, the reaction was stirred overnight, and the reaction was completed by EtOAc. EtOAc (EtOAc) (EtOAc) Silica gel column chromatography (ethyl acetate: petroleum ether = 1 : 3) gave Compound 8c (1.00 g, 66.2%). 1 H NMR (400MHz, CDCl 3 ) δ: 3.80-3.73 (m, 1H), 3.62-3.57 (m, 2H), 3.21 (s, 3H), 2.37-2.30 (m, 2H), 2.08-2.02 (m , 1H), 1.67-1.60 (m, 2H).

化合物8d的制备：将化合物8c(1.00g，8.61mmol)溶于15mL二氯甲烷中，加入三乙胺(2.61g，25.83mmol)，冰浴条件下缓慢滴加甲磺酰氯(1.48g，12.92mmol)，滴毕，搅拌2小时，TLC监测反应完毕，加入50mL饱和碳酸氢钠溶液，二氯甲烷萃取(30mL×3)，合并有机相，饱和食盐水洗涤，无水硫酸钠干燥，浓缩蒸干备用。Preparation of compound 8d: Compound 8c (1.00 g, 8.61 mmol) was dissolved in 15 mL of dichloromethane, triethylamine (2.61 g, 25.83 mmol) was added, and methanesulfonyl chloride (1.48 g, 12.92) was slowly added dropwise in an ice bath. After the completion of the dropwise addition, the mixture was stirred for 2 hours, and the reaction was completed by TLC. 50 mL of saturated sodium bicarbonate solution and dichloromethane (30mL×3) were added, and the organic phase was combined, washed with saturated brine, dried over anhydrous sodium sulfate and evaporated. Dry spare.

化合物8e的制备：将化合物10(0.46g，1.29mmol)，化合物8d(0.50g，2.58mmol)和碳酸钾(0.53g，3.87mmol)加入10mL无水N，N-二甲基甲酰胺中，加热至90℃反应5小时。TLC监测反应完毕，将反应液倒入水中，乙酸乙酯(30mL×3)萃取，合并有机相，饱和食盐水洗涤，无水硫酸钠干燥，浓缩蒸干得化合物8e粗品0.50g。Preparation of compound 8e: Compound 10 (0.46 g, 1.29 mmol), compound 8d (0.50 g, 2.58 mmol) and potassium carbonate (0.53 g, 3.87 mmol) were added to 10 mL of anhydrous N,N-dimethylformamide. The reaction was heated to 90 ° C for 5 hours. After the reaction was completed by TLC, the reaction mixture was poured into water, ethyl acetate (30 mL×3), and the organic phase was combined, washed with saturated brine, dried over anhydrous sodium sulfate and evaporated to dryness.

化合物I-8的制备：将化合物8e(0.50g，1.10mmol)溶于15mL四氢呋喃中，加入1N氢氧化钠水溶液(6.6mL，6.60mmol)，在35℃条件下反应3小时，TLC监测反应完毕。用1N盐酸调至pH＝1-2，有固体析出，过滤，用乙醚和乙醇重结晶得化合物I-8产品(270mg，57.3％)。1H NMR(400MHz，CDCl3)δ：8.44(s，1H)，7.17(s，1H)，7.05(s，1H)，6.72(s，1H)，4.07-4.05(m，2H)，3.91(s，3H)，3.88-3.82(m，2H)，3.36-3.31(dd，J1＝6Hz，J2＝16.8Hz，1H)，3.25(s，3H)，3.08-3.03(m，1H)，2.53-2.48(m，2H)，2.42-2.38(m，1H)，1.83-1.77(m，3H)，0.94(d，J＝6.4Hz，3H)，0.82(d，J＝6.8Hz，3H)；ESI-MS m/z 428.2(M+H)+。Preparation of compound I-8: Compound 8e (0.50 g, 1.10 mmol) was dissolved in 15 mL of tetrahydrofuran, 1N aqueous sodium hydroxide (6.6 mL, 6.60 mmol) was added, and the reaction was carried out at 35 ° C for 3 hours, and the reaction was completed by TLC. . The mixture was adjusted to pH = 1-2 with 1N hydrochloric acid, and then solid crystals, filtered, and recrystallized from diethyl ether and ethyl alcohol to give the product of compound I-8 (270 mg, 57.3%). 1 H NMR (400MHz, CDCl 3 ) δ: 8.44 (s, 1H), 7.17 (s, 1H), 7.05 (s, 1H), 6.72 (s, 1H), 4.07-4.05 (m, 2H), 3.91 ( s, 3H), 3.88-3.82 (m, 2H), 3.36-3.31 (dd, J 1 = 6 Hz, J 2 = 16.8 Hz, 1H), 3.25 (s, 3H), 3.08-3.03 (m, 1H), 2.53-2.48 (m, 2H), 2.42 - 2.38 (m, 1H), 1.83-1.77 (m, 3H), 0.94 (d, J = 6.4 Hz, 3H), 0.82 (d, J = 6.8 Hz, 3H) ;ESI-MS m/z 428.2 (M+H) + .

实施例13.制备6-异丙基-10-甲氧基-9-((3-甲氧基环丁基)甲氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-8-rac)Example 13. Preparation of 6-isopropyl-10-methoxy-9-((3-methoxycyclobutyl)methoxy)-2-oxo-6,7-dihydro-2H-pyridine And [2,1-a]isoquinoline-3-carboxylic acid (I-8-rac)

Figure PCTCN2017094946-appb-000024
Figure PCTCN2017094946-appb-000024

Figure PCTCN2017094946-appb-000025
Figure PCTCN2017094946-appb-000025

按照实施例12的方法，得到化合物I-8-rac.1H NMR(400MHz，CDCl3)δ：8.44(s，1H)，7.17(s，1H)，7.05(s，1H)，6.72(s，1H)，4.07-4.05(m，2H)，3.91(s，3H)，3.88-3.82(m，2H)，3.36-3.31(dd，J1＝6Hz，J2＝16.8Hz，1H)，3.25(s，3H)，3.08-3.03(m，1H)，2.53-2.48(m，2H)，2.42-2.38(m，1H)，1.83-1.77(m，3H)，0.94(d，J＝6.4Hz，3H)，0.82(d，J＝6.8Hz，3H)；ESI-MS m/z 428.2(M+H)+。The compound I-8-rac. 1 H NMR (400 MHz, CDCl 3 ) δ: 8.44 (s, 1H), 7.17 (s, 1H), 7.05 (s, 1H), 6.72 (s) , 1H), 4.07-4.05 (m, 2H), 3.91 (s, 3H), 3.88-3.82 (m, 2H), 3.36-3.31 (dd, J 1 = 6 Hz, J 2 = 16.8 Hz, 1H), 3.25 (s, 3H), 3.08-3.03 (m, 1H), 2.53-2.48 (m, 2H), 2.42-2.38 (m, 1H), 1.83-1.77 (m, 3H), 0.94 (d, J = 6.4 Hz) , 3H), 0.82 (d, J = 6.8 Hz, 3H); ESI-MS m/z 428.2 (M+H) + .

实施例14.制备(S)-6-异丙基-10-甲氧基-2-氧代-9-((4-氧代戊基)氧基)-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-9)Example 14. Preparation of (S)-6-isopropyl-10-methoxy-2-oxo-9-((4-oxopentyl)oxy)-6,7-dihydro-2H- Pyrido[2,1-a]isoquinoline-3-carboxylic acid (I-9)

Figure PCTCN2017094946-appb-000026
Figure PCTCN2017094946-appb-000026

化合物9a的制备：将化合物10(0.36g，1.0mmol)溶于10mL四氢呋喃中，加入5-羟基-2-戊酮(0.20g，2.0mmol)和三苯基膦(0.53g，2.0mmol)。氮气置换4次，置于冰水浴中搅拌30分钟，将偶氮二甲酸二乙酯(0.35g，2.0mmol)滴入上述体系中，室温搅拌16小时。加入硅胶拌样，柱层析纯化得产物(0.25g，57.0％)。Preparation of compound 9a: Compound 10 (0.36 g, 1.0 mmol) was dissolved in 10 mL of tetrahydrofuran, and 5-hydroxy-2-pentanone (0.20 g, 2.0 mmol) and triphenylphosphine (0.53 g, 2.0 mmol). The mixture was replaced with nitrogen for 4 times, stirred in an ice water bath for 30 minutes, and diethyl azodicarboxylate (0.35 g, 2.0 mmol) was added dropwise to the above system, and stirred at room temperature for 16 hours. The mixture was added to a silica gel and purified by column chromatography to yield (0.25 g, 57.0%).

化合物I-9的制备：将化合物9a(0.25g，0.57mmol)溶于10mL四氢呋喃和5mL水中，加入1N氢氧化钠水溶液(2.3mL，2.3mmol)，在35℃条件下反应3小时，TLC监测反应完毕。用1N盐酸调至pH＝1-2，加入20mL二氯甲烷，分液，水层用二氯甲烷萃取(30mL×2)，合并有机层，用饱和氯化钠洗，无水硫酸钠干燥，过滤后浓缩得粗品300mg。将该粗品溶于2mL乙醇中，加热回流20分钟后冷却至室温，加入30mL乙醚，搅拌30分钟。产品抽滤烘干，即得化合物I-9(130mg，55.1％)。1H NMR(400MHz，CDCl3)δ：16.01(s，1H)，8.45(s，1H)，7.16(s，1H)，7.05(s，1H)，6.76(s，1H)，4.08-4.14(m，2H)，3.91(s，3H)，3.85-3.89(m，1H)，3.30-3.36(m，1H)，3.06(d，J＝15.6Hz，1H)，2.69(t，J＝6.8Hz，2H)，2.78(s，3H)，2.10-2.15(m，2H)，1.81(m，1H)，2.27(m，2H)，1.58(m，2H)，0.94(d，J＝6.4Hz，3H)，0.81(d，J＝6.4Hz，3H)；ESI-MS m/z 414.2(M+H)+。Preparation of compound I-9: Compound 9a (0.25 g, 0.57 mmol) was dissolved in 10 mL of tetrahydrofuran and 5 mL of water, and 1N aqueous sodium hydroxide (2.3 mL, 2.3 mmol) was added and reacted at 35 ° C for 3 hours, TLC monitoring The reaction is completed. The mixture was adjusted to pH 1-2 with 1N EtOAc. EtOAc (EtOAc m. After filtration, it was concentrated to give a crude product (300 mg). The crude product was dissolved in 2 mL of ethanol, heated to reflux for 20 min and then cooled to room temperature. The product was dried by suction filtration to give Compound I-9 (130 mg, 55.1%). 1 H NMR (400MHz, CDCl 3 ) δ: 16.01 (s, 1H), 8.45 (s, 1H), 7.16 (s, 1H), 7.05 (s, 1H), 6.76 (s, 1H), 4.08-4.14 ( m, 2H), 3.91 (s, 3H), 3.85-3.89 (m, 1H), 3.30-3.36 (m, 1H), 3.06 (d, J = 15.6 Hz, 1H), 2.69 (t, J = 6.8 Hz) , 2H), 2.78 (s, 3H), 2.10-2.15 (m, 2H), 1.81 (m, 1H), 2.27 (m, 2H), 1.58 (m, 2H), 0.94 (d, J = 6.4 Hz, 3H), 0.81 (d, J = 6.4 Hz, 3H); ESI-MS m/z 414.2 (M+H) + .

实施例15.制备-6-异丙基-10-甲氧基-2-氧代-9-((4-氧代戊基)氧基)-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-9-rac)Example 15. Preparation of -6-isopropyl-10-methoxy-2-oxo-9-((4-oxopentyl)oxy)-6,7-dihydro-2H-pyridine and [ 2,1-a]isoquinoline-3-carboxylic acid (I-9-rac)

Figure PCTCN2017094946-appb-000027
Figure PCTCN2017094946-appb-000027

按照实施例14的方法，得到化合物I-9-rac。1H NMR(400MHz，CDCl3)δ：16.01(s，1H)，8.45(s，1H)，7.16(s，1H)，7.05(s，1H)，6.76(s，1H)，4.08-4.14(m，2H)，3.91(s，3H)，3.85-3.89(m，1H)，3.30-3.36(m，1H)，3.06(d，J＝15.6Hz，1H)，2.69(t，J＝6.8Hz，2H)，2.78(s，3H)，2.10-2.15(m，2H)，1.81(m，1H)，2.27(m，2H)，1.58(m，2H)，0.94(d，J＝6.4Hz，3H)，0.81(d，J＝6.4Hz，3H)；ESI-MS m/z 414.2(M+H)+。According to the method of Example 14, the compound I-9-rac was obtained. 1 H NMR (400MHz, CDCl 3 ) δ: 16.01 (s, 1H), 8.45 (s, 1H), 7.16 (s, 1H), 7.05 (s, 1H), 6.76 (s, 1H), 4.08-4.14 ( m, 2H), 3.91 (s, 3H), 3.85-3.89 (m, 1H), 3.30-3.36 (m, 1H), 3.06 (d, J = 15.6 Hz, 1H), 2.69 (t, J = 6.8 Hz) , 2H), 2.78 (s, 3H), 2.10-2.15 (m, 2H), 1.81 (m, 1H), 2.27 (m, 2H), 1.58 (m, 2H), 0.94 (d, J = 6.4 Hz, 3H), 0.81 (d, J = 6.4 Hz, 3H); ESI-MS m/z 414.2 (M+H) + .

实施例16.制备(S)-9-(环戊氧基)-6-异丙基-10-甲氧基-2-氧代-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-10)Example 16. Preparation of (S)-9-(cyclopentyloxy)-6-isopropyl-10-methoxy-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylic acid (I-10)

Figure PCTCN2017094946-appb-000028
Figure PCTCN2017094946-appb-000028

化合物10a的制备：将化合物10(180mg，0.50mmol)溶于20mLN，N-二甲基甲酰胺中，加入溴代环戊烷(100mg，1.5mmol)和碳酸钾(138mg，1.0mmol)。在85℃条件下反应3小时，冷却至室温，加入60mL水和50mL乙酸乙酯，分液，水层用乙酸乙酯萃取(30mL×3)，合并有机层，用饱和氯化钠洗一次，浓缩得化合物10a粗品300mg，直接用于下一步反应。Preparation of Compound 10a: Compound 10 (180 mg, 0.50 mmol) was dissolved in 20 mL of N,N-dimethylformamide, bromocyclopentane (100 mg, 1.5 mmol) and potassium carbonate (138 mg, 1.0 mmol). The reaction was carried out at 85 ° C for 3 hours, cooled to room temperature, 60 mL of water and 50 mL of ethyl acetate were added, and the mixture was separated, and the aqueous layer was extracted with ethyl acetate (30 mL × 3). The crude compound 10a was concentrated to give 300 mg, which was directly used for the next reaction.

化合物I-10的制备：将化合物10a(300mg，0.71mmol)溶于10mL四氢呋喃和5mL水中，加入1N氢氧化钠水溶液(2.8mL，2.8mmol)，室温搅拌16小时，TLC监测反应完毕。用1N盐酸调至pH＝1-2，加入20mL二氯甲烷，分液，水层用二氯甲烷萃取(30mL×2)，合并有机层，用饱和氯化钠洗，无水硫酸钠干燥，过滤后浓缩得粗品300mg。将该粗品溶于2mL乙醇中，加热回流20分钟后冷却至室温，加入30mL乙醚，搅拌30分钟。抽滤，烘干得化合物I-10(110mg，39.4％)。1H NMR(400MHz，CDCl3)δ：8.44(s，1H)，7.15(s，1H)，7.04(s，1H)，6.72(s，1H)，4.82-4.87(m，1H)，3.90(s，3H)，3.84-3.89(m，1H)，3.31-3.36(m，1H)，3.03-3.07(d，J＝16Hz，1H)，1.81-2.00(m，6H)，1.61-1.67(m，4H)，0.94(d，J＝6.8Hz，3H)，0.82(d，J＝6.8Hz，3H)；ESI-MS m/z 398.2(M+H)+。Preparation of Compound I-10: Compound 10a (300 mg, 0.71 mmol) was dissolved in 10 mL of THF and 5 mL of water, and 1N sodium hydroxide aqueous solution (2.8 mL, 2.8 mmol) was added and stirred at room temperature for 16 hours. The mixture was adjusted to pH 1-2 with 1N EtOAc. EtOAc (EtOAc m. After filtration, it was concentrated to give a crude product (300 mg). The crude product was dissolved in 2 mL of ethanol, heated to reflux for 20 min and then cooled to room temperature. Filtration and drying gave Compound I-10 (110 mg, 39.4%). 1 H NMR (400MHz, CDCl 3 ) δ: 8.44 (s, 1H), 7.15 (s, 1H), 7.04 (s, 1H), 6.72 (s, 1H), 4.82-4.87 (m, 1H), 3.90 ( s, 3H), 3.84-3.89 (m, 1H), 3.31-3.36 (m, 1H), 3.03-3.07 (d, J = 16 Hz, 1H), 1.81-2.00 (m, 6H), 1.61-1.67 (m , 4H), 0.94 (d, J = 6.8 Hz, 3H), 0.82 (d, J = 6.8 Hz, 3H); ESI-MS m/z 398.2 (M+H) + .

实施例17.制备9-(环戊氧基)-6-异丙基-10-甲氧基-2-氧代-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-10-rac)Example 17. Preparation of 9-(cyclopentyloxy)-6-isopropyl-10-methoxy-2-oxo-6,7-dihydro-2H-pyrido[2,1-a] Quinoline-3-carboxylic acid (I-10-rac)

Figure PCTCN2017094946-appb-000029
Figure PCTCN2017094946-appb-000029

按照实施例16的方法，得到I-10-rac。1HNMR(400MHz，CDCl3)δ：8.44(s，1H)，7.15(s，1H)，7.04(s，1H)，6.72(s，1H)，4.82-4.87(m，1H)，3.90(s，3H)，3.84-3.89(m，1H)，3.31-3.36(m，1H)，3.03-3.07(d，J＝16Hz，1H)，1.81-2.00(m，6H)，1.61-1.67(m，4H)，0.94(d，J＝6.8Hz，3H)，0.82(d，J＝6.8Hz，3H)；ESI-MS m/z 398.2(M+H)+。According to the method of Example 16, I-10-rac was obtained. 1 H NMR (400 MHz, CDCl 3 ) δ: 8.44 (s, 1H), 7.15 (s, 1H), 7.04 (s, 1H), 6.72 (s, 1H), 4.82-4.87 (m, 1H), 3.90 (s) , 3H), 3.84-3.89 (m, 1H), 3.31-3.36 (m, 1H), 3.03-3.07 (d, J = 16 Hz, 1H), 1.81-2.00 (m, 6H), 1.61-1.67 (m, 4H), 0.94 (d, J = 6.8 Hz, 3H), 0.82 (d, J = 6.8 Hz, 3H); ESI-MS m/z 398.2 (M+H) + .

实施例18.制备(S)-9-(环丙基乙炔基)-6-异丙基-10-甲氧基-2-氧代-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-11)Example 18. Preparation of (S)-9-(cyclopropylethynyl)-6-isopropyl-10-methoxy-2-oxo-6,7-dihydro-2H-pyrido[2, 1-a]isoquinoline-3-carboxylic acid (I-11)

Figure PCTCN2017094946-appb-000030
Figure PCTCN2017094946-appb-000030

化合物11a的制备：化合物10(0.50g，1.40mmol)溶于10mL二氯甲烷，加入三乙胺(0.42g，4.2mmol)，滴加N-苯基双(三氟甲烷磺酸亚胺)(0.75g，2.1mmol)的二氯甲烷溶液(6mL)，约3分钟滴完，在0℃搅拌30分钟后回到室温，在室温下搅拌2小时，真空浓缩，粗品经过柱层析纯化得到化合物11a(0.60g，87.6％)。Preparation of compound 11a: Compound 10 (0.50 g, 1.40 mmol) was dissolved in 10 mL dichloromethane, triethylamine (0.42 g, 4.2 mmol) was added, and N-phenylbis(trifluoromethanesulfonimide) was added dropwise ( A solution of 0.75 g (2.1 mmol) in dichloromethane (6 mL) was added dropwise over a period of 3 minutes, stirred at 0 ° C for 30 min, then returned to room temperature, stirred at room temperature for 2 h, concentrated in vacuo and purified 11a (0.60g, 87.6%).

化合物11c的制备：化合物11b(1.32g，20mmol)溶于40mL无水四氢呋喃中，氮气置换3次，冷却至-78℃，搅拌1小时，加入硼酸三甲酯(4.67g，45mmol)，在-78℃搅拌1小时后回到-20℃，在-20℃搅拌1小时，加入饱和氟氢化钾饱和溶液(14.06g，0.18mol)，剧烈搅拌1小时，回到室温搅拌1小时后，溶剂在减压条件除去，得到白色固体，真空干燥，产品溶于40mL热丙酮，过滤，滤液旋干，固体溶于5mL丙酮，加热至回流，加入50mL乙醚，冷却至0℃，过滤，滤饼真空干燥得到化合物11c(1.72g，50％)。Preparation of compound 11c: Compound 11b (1.32 g, 20 mmol) was dissolved in 40 mL of anhydrous THF. After stirring at 78 ° C for 1 hour, return to -20 ° C, stir at -20 ° C for 1 hour, add saturated potassium fluorohydride solution (14.06 g, 0.18 mol), stir vigorously for 1 hour, return to room temperature and stir for 1 hour, the solvent is After removal under reduced pressure, a white solid was obtained, dried in vacuo, evaporated, evaporated, evaporated, evaporated, evaporated, evaporated, evaporated, evaporated, evaporated, evaporated, evaporated. Compound 11c (1.72 g, 50%) was obtained.

化合物11d的制备：将化合物11a(0.25g，0.51mmol)，化合物11c(0.12g，0.66mmol)，Pd(dppf)Cl2(37mg，0.051 mmol)和碳酸钠(0.11g，1.02mmol)加入到12mL甲基叔丁基醚和3mL水的混合溶剂中，氮气置换3次，加热至100℃反应3小时，TLC显示反应完全，反应液直接柱层析得到化合物11d(98mg，47.1％)。Preparation of compound 11d: Compound 11a (0.25 g, 0.51 mmol), compound 11c (0.12 g, 0.66 mmol), Pd (dppf) Cl 2 (37 mg, 0.051 mmol) and sodium carbonate (0.11 g, 1.02 mmol) In a mixed solvent of 12 mL of methyl tert-butyl ether and 3 mL of water, the mixture was replaced with nitrogen three times, and heated to 100 ° C for 3 hours. TLC showed the reaction was completed, and the reaction mixture was directly subjected to column chromatography to give compound 11d (98 mg, 47.1%).

化合物I-11的制备：化合物11d(98mg，0.24mmol)溶于3mL四氢呋喃，加入1N氢氧化钠水溶液(1.3mL，1.3mmol)，室温反应18小时。加入1N盐酸调节至pH＝1-2，然后二氯甲烷萃取(30mL×2)，合并有机层，无水硫酸钠干燥，过滤，滤液旋干，粗品用乙酸乙酯与甲基叔丁基醚重结晶得到化合物I-11(32mg，35.3％)。1H NMR(400MHz，CDCl3)δ：8.46(s，1H)，7.25(s，1H)，7.16-7.11(m，2H)，3.93(s，3H)，3.88-3.82(m，1H)，3.51-3.44(m，1H)，3.30-3.21(m，1H)，3.10-3.03(m，1H)，1.57-1.49(m，1H)，1.29-1.23(m，1H)，0.96-0.78(m，10H)；ESI-MS m/z 378.2(M+H)+。Preparation of Compound I-11: Compound 11d (98 mg, 0.24 mmol) was dissolved in 3 mL of THF, and 1N aqueous sodium hydroxide (1.3 mL, 1.3 mmol). Add 1N hydrochloric acid to adjust to pH = 1-2, then extract with dichloromethane (30 mL × 2), the organic layer is combined, dried over anhydrous sodium sulfate, filtered, and evaporated. Recrystallization gave Compound I-11 (32 mg, 35.3%). 1 H NMR (400MHz, CDCl 3 ) δ: 8.46 (s, 1H), 7.25 (s, 1H), 7.16-7.11 (m, 2H), 3.93 (s, 3H), 3.88-3.82 (m, 1H), 3.51-3.44 (m, 1H), 3.30-3.21 (m, 1H), 3.10-3.03 (m, 1H), 1.57-1.49 (m, 1H), 1.29-1.23 (m, 1H), 0.96-0.78 (m) , 10H); ESI-MS m/z 378.2 (M+H) + .

实施例19.制备9-(环丙基乙炔基)-6-异丙基-10-甲氧基-2-氧代-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-11-rac)Example 19. Preparation of 9-(cyclopropylethynyl)-6-isopropyl-10-methoxy-2-oxo-6,7-dihydro-2H-pyrido[2,1-a] Isoquinoline-3-carboxylic acid (I-11-rac)

Figure PCTCN2017094946-appb-000031
Figure PCTCN2017094946-appb-000031

按照实施例18的方法，得到化合物I-11-rac。1H NMR(400MHz，CDCl3)δ：8.46(s，1H)，7.25(s，1H)，7.16-7.11(m，2H)，3.93(s，3H)，3.88-3.82(m，1H)，3.51-3.44(m，1H)，3.30-3.21(m，1H)，3.10-3.03(m，1H)，1.57-1.49(m，1H)，1.29-1.23(m，1H)，0.96-0.78(m，10H)；ESI-MS m/z 378.2(M+H)+。According to the method of Example 18, the compound I-11-rac was obtained. 1 H NMR (400MHz, CDCl 3 ) δ: 8.46 (s, 1H), 7.25 (s, 1H), 7.16-7.11 (m, 2H), 3.93 (s, 3H), 3.88-3.82 (m, 1H), 3.51-3.44 (m, 1H), 3.30-3.21 (m, 1H), 3.10-3.03 (m, 1H), 1.57-1.49 (m, 1H), 1.29-1.23 (m, 1H), 0.96-0.78 (m) , 10H); ESI-MS m/z 378.2 (M+H) + .

实施例20.制备(S)-6-异丙基-10-甲氧基-9-(4-甲氧基丁-1-炔-1-基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-α]异喹啉-3-羧酸(I-12)Example 20. Preparation of (S)-6-isopropyl-10-methoxy-9-(4-methoxybut-1-yn-1-yl)-2-oxo-6,7-di Hydrogen-2H-pyrido[2,1-α]isoquinoline-3-carboxylic acid (I-12)

Figure PCTCN2017094946-appb-000032
Figure PCTCN2017094946-appb-000032

化合物12b的制备：化合物12a(1.30g，15.0mmol)溶于30mL无水四氢呋喃中，氮气置换3次，冷却至-78℃，搅拌1小时，加入硼酸三甲酯(2.34g，22.0mmol)，在-78℃搅拌1小时后回到-20℃，在-20℃搅拌1小时，加入氟氢化钾饱和溶液(7.03g，90.0mmol)，剧烈搅拌1小时，回到室温搅拌1小时后，溶剂在减压条件去除，得到白色固体，真空干燥，产品溶于40mL热丙酮，过滤，滤液旋干，固体溶于5mL丙酮，加热至回流，加入50mL乙醚，冷却至0℃，过滤，滤饼真空干燥得到固体(0.32g，11.2％)，即为化合物12b。Preparation of compound 12b: Compound 12a (1.30 g, 15.0 mmol) was dissolved in 30 mL of anhydrous THF. After stirring at -78 ° C for 1 hour, return to -20 ° C, stir at -20 ° C for 1 hour, add potassium hydride saturated solution (7.03 g, 90.0 mmol), stir vigorously for 1 hour, return to room temperature and stir for 1 hour, solvent After removal under reduced pressure, a white solid was obtained, which was dried in vacuo. The product was dissolved in 40 mL of hot acetone, filtered, and the filtrate was evaporated to dryness. The solid was dissolved in 5 mL of acetone, heated to reflux, 50 mL diethyl ether was added, cooled to 0 ° C, filtered, vacuum cake Drying gave a solid (0.32 g, 11.2%) as compound 12b.

化合物12c的制备：将化合物11a(120mg，0.25mmol)，化合物12b(71mg，0.37mmol)，Pd(dppf)Cl2(18mg，0.025mmol)，碳酸钠(78mg，0.74mmol)加入到9mL乙二醇二甲醚和2mL水中，氮气置换3次，加热至100℃，反应3小时，TLC显示反应完全，反应液直接柱层析得到化合物12c(55mg，52.0％)。Preparation of compound 12c: Compound 11a (120 mg, 0.25 mmol), compound 12b (71 mg, 0.37 mmol), Pd (dppf) Cl 2 (18 mg, 0.025 mmol), sodium carbonate (78 mg, 0.74 mmol) Alcohol dimethyl ether and 2 mL of water were replaced with nitrogen three times, heated to 100 ° C, and reacted for 3 hours. TLC showed the reaction was completed, and the reaction mixture was directly subjected to column chromatography to give compound 12c (55 mg, 52.0%).

化合物I-12的制备：往化合物12c(55mg，0.13mmol)的THF(3mL)溶液中加入1N氢氧化钠水溶液(1.28mL，1.28mmol)，室温反应18小时。加入1N盐酸调节至pH＝1-2，二氯甲烷萃取(30mL×2)，合并有机层，无水硫酸钠干燥，过滤，滤液旋干，粗品用制备色谱纯化得到化合物I-12(11mg，21.4％)。1H NMR(400MHz，CDCl3)δ：8.49-8.48(m，1H)，7.33(s，1H)，7.18-7.17(m，2H)，3.96(s，3H)，3.92-3.85(m，1H)，3.64(t，J＝7.2Hz，2H)，3.44(s，3H)， 3.32-3.25(m，1H)，3.14-3.07(m，1H)，2.80(t，J＝6.4Hz，2H)，0.95(d，J＝6.4Hz，3H)，0.91(d，J＝6.4Hz，3H)；ESI-MS m/z 396.2(M+H)+。Preparation of Compound I-12: To a solution of EtOAc (3 mL, EtOAc. After adding 1N hydrochloric acid, the mixture was adjusted to pH = 1-2, dichloromethane (30 mL × 2), the organic layer was combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. 21.4%). 1 H NMR (400MHz, CDCl 3 ) δ: 8.49-8.48 (m, 1H), 7.33 (s, 1H), 7.18-7.17 (m, 2H), 3.96 (s, 3H), 3.92-3.85 (m, 1H ), 3.64 (t, J = 7.2 Hz, 2H), 3.44 (s, 3H), 3.32-3.25 (m, 1H), 3.14 - 3.07 (m, 1H), 2.80 (t, J = 6.4 Hz, 2H) , 0.95 (d, J = 6.4 Hz, 3H), 0.91 (d, J = 6.4 Hz, 3H); ESI-MS m/z 396.2 (M+H) + .

实施例21.制备(S)-6-异丙基-10-甲氧基-9-((1s，3R)-3-甲氧基环丁氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-α]异喹啉-3-羧酸(I-19)Example 21. Preparation of (S)-6-isopropyl-10-methoxy-9-((1s,3R)-3-methoxycyclobutoxy)-2-oxo-6,7- Dihydro-2H-pyrido[2,1-α]isoquinoline-3-carboxylic acid (I-19)

Figure PCTCN2017094946-appb-000033
Figure PCTCN2017094946-appb-000033

化合物19b：化合物19a(1.60g，8.99mmol)，对硝基苯甲酸(1.50g，8.99mmol)和三苯基膦(4.24g，16.18mmol)混合溶于10mL无水THF，氮气置换3次后，冰水浴下滴加偶氮二甲酸二乙酯(2.35g，13.48mmol)到体系中，室温搅拌19小时，旋干得到粗品，直接柱层析(PE→PE∶EA＝50∶1)得到固体2.43g，收率：83.0％。Compound 19b: Compound 19a (1.60 g, 8.99 mmol), p-nitrobenzoic acid (1.50 g, 8.99 mmol) and triphenylphosphine (4.24 g, 16.18 mmol) were dissolved in 10 mL of anhydrous THF. Ethyl azodicarboxylate (2.35 g, 13.48 mmol) was added dropwise to the system under ice-water bath, stirred at room temperature for 19 hours, and dried to give a crude product, which was obtained by column chromatography (PE: PE: EA = 50:1). Solid 2.43 g, yield: 83.0%.

化合物19c：化合物31b(2.43g，7.43mmol)溶于THF(3ml)和甲醇(1ml)，加入氢氧化锂(1.873g，44.6mmol)的水溶液(10mL)，室温搅拌4小时，减压浓缩有机溶剂，乙酸乙酯萃取(100ml×3)，合并乙酸乙酯层，依次用水和饱和食盐水洗，无水硫酸钠干燥，过滤，滤液真空浓缩得到粗产品1.11g。Compound 19c: Compound 31b (2.43 g, 7.43 mmol) eluted eluted eluted The solvent was extracted with EtOAc (EtOAc mEtOAcEtOAcEtOAc.

化合物19d：化合物3c(1.11g，6.21mmol)溶于15mL无水THF，加入氢化钠(0.30g，12.42mmol)，搅拌0.5小时后，加入碘甲烷(1.15g，8.07mmol)，加料完毕后，室温搅拌2小时，然后加入10mL水淬灭，加入乙酸乙酯萃取(30ml×3)，合并乙酸乙酯层，无水硫酸钠干燥，滤液直接拌硅胶柱层析(PE∶EA＝30∶1)得到1.10g化合物。收率：92.1％。Compound 19d: Compound 3c (1.11 g, 6.21 mmol) was dissolved in 15 mL of dry THF, and then sodium hydride (0.30 g, 12.42 mmol) was added, and after stirring for 0.5 hour, iodomethane (1.15 g, 8.07 mmol) was added, after the addition, After stirring at room temperature for 2 hours, it was then quenched with 10 mL of water and extracted with ethyl acetate (30 ml×3). The ethyl acetate layer was combined and dried over anhydrous sodium sulfate. The filtrate was directly applied to silica gel column chromatography (PE:EA=30:1) ) 1.10 g of compound was obtained. Yield: 92.1%.

化合物19e：化合物31d(1.10g，5.72mmol)溶于10mL甲醇，加入Pd/C(0.325g)，加入1滴浓盐酸，氢气真空置换3次，室温加氢18小时，过滤，滤液真空浓缩得到产品0.56g。收率：95.9％。Compound 19e: Compound 31d (1.10 g, 5.72 mmol) was dissolved in 10 mL of methanol, Pd/C (0.325 g) was added, 1 drop of concentrated hydrochloric acid was added, and hydrogen was replaced by vacuum three times, and hydrogenated at room temperature for 18 hours, filtered, and the filtrate was concentrated in vacuo. The product is 0.56g. Yield: 95.9%.

化合物19f：化合物31e(0.560g，5.49mmol)溶于8mL二氯甲烷中，加入三乙胺(1.39g，13.72mmol)，冰水浴下缓慢加入甲磺酰氯(0.94g，8.23mmol)，混合物室温搅拌2小时，旋干，加入30mL水，乙酸乙酯萃取(50ml×2)，有机相合并，依次用水洗和饱和食盐水洗，无水硫酸钠干燥，滤液旋干，得到粗产品0.910g。Compound 19f: Compound 31e (0.560 g, 5.49 mmol) was dissolved in dichloromethane (3 mL), triethylamine (1.39 g, 13.72 mmol), and methanesulfonyl chloride (0.94 g, 8.23 mmol). After stirring for 2 hours, the mixture was stirred with EtOAc EtOAc (EtOAc)EtOAc.

化合物19g：化合物10(0.30g，0.84mmol)和化合物19f(0.23g，1.26mmol)溶于6mL DMF中，加入碳酸钾(0.35g，2.52mmol)，在90℃反应18小时。反应结束后倒入水中，乙酸乙酯萃取(50ml×4)，合并乙酸乙酯层，依次用水洗和饱和食盐水洗，无水硫酸钠干燥，过滤，滤液真空浓缩，得到粗产品0.42g。Compound 19g: Compound 10 (0.30 g, 0.84 mmol) and Compound 19f (0.23 g, 1.26 mmol) were dissolved in 6 mL of DMF, and potassium carbonate (0.35 g, 2.52 mmol) was added and reacted at 90 ° C for 18 hours. After completion of the reaction, the mixture was poured into EtOAc EtOAc m.

化合物I-19：化合物19g(0.42g，0.95mmol)溶于5mL THF中，加入10％氢氧化钠水溶液(0.30g，7.50mmol)，反应18小时，用1N盐酸将pH调至1-2，用二氯甲烷萃取(40mL×2)，合并有机层，用饱和氯化钠洗，干燥浓缩后，粗品用乙酸乙酯与甲基叔丁基醚重结晶得到产品0.170g。1HNMR(400MHz，CDCl3)δ：8.47(s，1H)，7.18(s，1H)，7.07(s，1H)，6.59(s，1H)，4.46-4.37(m，1H)，3.91-3.85(m，1H)，3.75-3.66(m，1H)，3.37-3.31(m，1H)，3.29(s，3H)，3.07(d，J＝15.6Hz，1H)，2.98-2.91(m，2H)，2.32-2.20(m，2H)，1.87-1.79(m，1H)，0.95(d，J＝6.4Hz，3H)，0.84(d，J＝6.8Hz，3H).ESI-MS m/z 414.2(M+H)+ Compound I-19: Compound 19 g (0.42 g, 0.95 mmol) was dissolved in 5 mL of THF, and then 10% aqueous sodium hydroxide (0.30 g, 7.50 mmol) was reacted for 18 hours, and the pH was adjusted to 1-2 with 1N hydrochloric acid. The organic layer was combined with EtOAc (3 mL, EtOAc). 1 H NMR (400 MHz, CDCl 3 ) δ: 8.47 (s, 1H), 7.18 (s, 1H), 7.07 (s, 1H), 6.59 (s, 1H), 4.46 - 4.37 (m, 1H), 3.91-3.85 (m, 1H), 3.75-3.66 (m, 1H), 3.37-3.31 (m, 1H), 3.29 (s, 3H), 3.07 (d, J = 15.6 Hz, 1H), 2.98-2.91 (m, 2H) ), 2.32-2.20 (m, 2H), 1.87-1.79 (m, 1H), 0.95 (d, J = 6.4 Hz, 3H), 0.84 (d, J = 6.8 Hz, 3H). ESI-MS m/z 414.2(M+H) +

实施例22.制备(S)-9-环丁氧基-6-异丙基-10-甲氧基-2-氧代-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-20) Example 22. Preparation of (S)-9-cyclobutoxy-6-isopropyl-10-methoxy-2-oxo-6,7-dihydro-2H-pyrido[2,1-a Isoquinoline-3-carboxylic acid (I-20)

Figure PCTCN2017094946-appb-000034
Figure PCTCN2017094946-appb-000034

化合物20a：化合物10(0.100g，0.28mmol)和溴代环丁烷(0.075g，0.56mmol)溶于DMF(4ml)，加入碳酸钾(0.116g，0.84mmol)，在90℃反应18小时。反应结束后倒入水中，乙酸乙酯萃取(50ml×4)，合并乙酸乙酯层，依次用水洗和饱和食盐水洗，无水硫酸钠干燥，过滤，滤液真空浓缩，得到粗产品63mg。Yield：54.6％。Compound 20a: Compound 10 (0.100 g, 0.28 mmol) and bromocyclobutane (0.075 g, 0.56 mmol) were dissolved in DMF (4 ml), and potassium carbonate (0.116 g, 0.84 mmol) was added and reacted at 90 ° C for 18 hours. After the completion of the reaction, the mixture was poured into EtOAc EtOAc m. Yield: 54.6%.

化合物I-20：化合物20a(0.063g，0.15mmol)溶于3mL THF，加入10％氢氧化钠水溶液(0.200g，5mmol)，室温反应18小时，用1N盐酸将pH调至1-2，用二氯甲烷萃取(40mL×2)，合并有机层，用饱和氯化钠洗，干燥浓缩后，粗品用乙酸乙酯与甲基叔丁基醚重结晶得到产品15mg。收率：25.5％。1HNMR(400MHz，CDCl3)δ：8.50(d，J＝0.8Hz，1H)，7.16(s，1H)，7.06(s，1H)，6.57(s，1H)，4.77-4.67(m，1H)，4.64-4.48(m，2H)，3.92(s，3H)，3.41-3.26(m，1H)，3.09-2.98(m，1H)，2.56-2.44(m，2H)，2.37-3.20(m，2H)，1.97-1.85(m，2H)，1.75-1.68(m，1H)，0.93(d，J＝6.4Hz，3H)，0.81(d，J＝6.4Hz，3H).ESI-MS m/z 384.2(M+H)+ Compound I-20: Compound 20a (0.063 g, 0.15 mmol) was dissolved in 3 mL of THF, then 10% aqueous sodium hydroxide (0.200 g, 5 mmol) was added and allowed to react at room temperature for 18 hours, and the pH was adjusted to 1-2 with 1N hydrochloric acid. The organic layer was combined with EtOAc (3 mL). Yield: 25.5%. 1 HNMR (400MHz, CDCl3) δ : 8.50 (d, J = 0.8Hz, 1H), 7.16 (s, 1H), 7.06 (s, 1H), 6.57 (s, 1H), 4.77-4.67 (m, 1H) , 4.64-4.48 (m, 2H), 3.92 (s, 3H), 3.41-3.26 (m, 1H), 3.09-2.98 (m, 1H), 2.56-2.44 (m, 2H), 2.37-3.20 (m, 2H), 1.97-1.85 (m, 2H), 1.75-1.68 (m, 1H), 0.93 (d, J = 6.4 Hz, 3H), 0.81 (d, J = 6.4 Hz, 3H). ESI-MS m/ z 384.2(M+H) +

实施例23.制备10-氯-6-异丙基-2-氧代-9-(((R)-四氢呋喃-3-基)氧基)-6，7-二氢-2H-吡啶并[2，1-α]异喹啉-3-羧酸(I-22-rac)Example 23. Preparation of 10-chloro-6-isopropyl-2-oxo-9-(((R)-tetrahydrofuran-3-yl)oxy)-6,7-dihydro-2H-pyridine[ 2,1-α]isoquinoline-3-carboxylic acid (I-22-rac)

Figure PCTCN2017094946-appb-000035
Figure PCTCN2017094946-appb-000035

化合物22b：将化合物1(2.07g，10mmol0溶于20mL乙腈中，加入苄溴(2.05g，12mmol)和碳酸钾(2.76g，20mmol)。于80℃加热搅拌16小时，反应结束后加入50mL水和50mL乙酸乙酯，分液，水层用50mL乙酸乙酯萃取，合并有机层，用饱和食盐水洗，干燥后浓缩，柱层析纯化得产物3.00g。收率：100％。Compound 22b: Compound 1 (2.07 g, 10 mmol) was dissolved in 20 mL of acetonitrile, benzyl bromide (2.05 g, 12 mmol) and potassium carbonate (2.76 g, 20 mmol) were added. The mixture was stirred under heating at 80 ° C for 16 hours, and 50 mL of water was added after completion of the reaction. The mixture was separated with 50 mL of ethyl acetate. EtOAc was evaporated.

化合物22c：将化合物22b(3.00g，10mmol)溶于50mL THF中，加入Pd2(dba)3(185mg，0.2mmol)，Xantphos(234mg，0.4mmol)和叔丁醇钠(1.55g，16.2mmol)。氮气置换3次，加入4-甲基2-丁酮(1.74g，20.2mmol)，于60℃加热搅拌6小时。加入硅胶拌样，色谱柱纯化得产物2.70g。收率：89.4％。Compound 22c: Compound 22b (3.00 g, 10 mmol) was dissolved in 50 mL THF. Pd 2 (dba) 3 (185 mg, 0.2 mmol), Xantphos (234 mg, 0.4 mmol) and sodium tert-butoxide (1.55 g, 16.2 mmol) ). After replacing with nitrogen three times, 4-methyl-2-butanone (1.74 g, 20.2 mmol) was added, and the mixture was stirred under heating at 60 ° C for 6 hours. The mixture was added to a silica gel and the column was purified to give 2.70 g of the product. Yield: 89.4%.

化合物22d-rac：将化合物22c(2.70g，8.94mmol)溶于50mL甲醇中，加入醋酸铵(6.90g，89.4mmol)和氰基硼氢化钠(1.12g，17.88mmol)，室温搅拌16小时。浓缩甲醇，加入NaOH水溶液(1.40g氢氧化钠溶于60mL水)和50mLDCM，搅拌20分钟。分液，水层用DCM萃取(50mL×2)，合并有机层，用饱和氯化钠洗，无水硫酸钠干燥，浓缩得粗产品2.70g，直接用于下一步反应。 Compound 22d-rac: Compound 22c (2.70 g, 8. <RTI ID=0.0></RTI> </RTI> <RTIgt; </RTI> <RTIgt; </RTI> <RTIgt; </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> <RTIgt; Methanol was concentrated, and aqueous NaOH (1.40 g of sodium hydroxide dissolved in 60 mL of water) and 50 m LDCM were added and stirred for 20 minutes. The organic layer was combined with EtOAc (EtOAc).

化合物22e-rac：将化合物22d-rac(2.70g，8.94mmol)溶于30mL二氧六环中，加入甲酸(3.50g，44.7mmol)，加热回流3小时。冷却至室温，加入50mL饱和碳酸氢钠水溶液和50mL乙酸乙酯，分液，水层用乙酸乙酯萃取(50mL×2)，合并有机层，用饱和氯化钠洗，无水硫酸钠干燥，浓缩得粗品3.20g，直接用于下一步反应。Compound 22e-rac: Compound 22d-rac (2.70 g, 8.94 mmol) was dissolved in 30 mL of dioxane, and toluene (3.50 g, 44.7 mmol). After cooling to room temperature, 50 mL of a saturated aqueous solution of sodium bicarbonate and 50 mL of ethyl acetate was added, and the mixture was evaporated. The crude product was concentrated to 3.20 g and used directly for the next reaction.

化合物22f-rac：将化合物22e-rac(3.20g，8.86mmol)溶于50mL乙腈中，加入三氯氧磷(1.63g，10.64mmol)，室温搅拌16小时。将反应液缓慢倒入50mL水中，浓缩乙腈，用氨水将pH调至8-9，加入50mLDCM，分液，水层用DCM萃取(50mL×2)，合并有机层，用饱和氯化钠洗，浓缩，加入硅胶拌样，色谱柱纯化得产物1.80g。收率：64.8％。Compound 22f-rac: Compound 22e-rac (3.20 g, 8.86 mmol) was dissolved in 50 mL of acetonitrile, and phosphorus oxychloride (1.63 g, 10.64 mmol) was added and stirred at room temperature for 16 hours. The reaction mixture was slowly poured into 50 mL of water, acetonitrile was concentrated, the pH was adjusted to 8-9 with aqueous ammonia, 50 m LDCM was added, the mixture was separated, and the aqueous layer was extracted with DCM (50 mL×2), and the organic layer was combined and washed with saturated sodium chloride. Concentrated, added to silica gel and sampled, and purified by column chromatography to obtain 1.80 g of product. Yield: 64.8%.

化合物22g-rac：将化合物22f-rac(1.80g，5.74mmol)溶于20mL乙醇和3mL水中，加入2-乙氧亚甲基乙酰乙酸乙酯(3.20g，17.2mmol)，于80℃加热搅拌4小时。浓缩乙醇和水得粗品3.60g，直接用于下一步反应。Compound 22g-rac: Compound 22f-rac (1.80 g, 5.74 mmol) was dissolved in 20 mL of ethanol and 3 mL of water, and ethyl 2-ethoxymethyleneacetate (3.20 g, 17.2 mmol) was added and stirred at 80 ° C. 4 hours. Concentrated ethanol and water gave 3.60 g of crude material which was directly used for the next reaction.

化合物22h-rac：将化合物22g-rac(3.60g，7.96mmol)溶于40mL乙二醇二甲醚中，加入四氯苯醌(1.96g，7.96mmol)，置于55℃加热搅拌3小时。加入硅胶拌样，色谱柱纯化得产品2.20g。收率：61.2％。Compound 22h-rac: Compound 22g-rac (3.60 g, 7.96 mmol) was dissolved in 40 mL of ethylene glycol dimethyl ether, and tetrachlorophenylhydrazine (1.96 g, 7.96 mmol) was added, and the mixture was heated and stirred at 55 ° C for 3 hours. The mixture was added to a silica gel and purified by a column to obtain 2.20 g of a product. Yield: 61.2%.

化合物22i-rac：将化合物22h-rac(2.10g，4.65mmol)溶于30mL干燥的二氯甲烷中，置于冰水浴中冷却，缓慢滴加三溴化硼(2.33g，9.29mmol)，冷却搅拌2小时。将反应液缓慢倒入50mL冰水中，加入50mL二氯甲烷，分液，水层用二氯甲烷萃取(50mL×2)，合并有机层，依次用饱和碳酸氢钠和饱和食盐水洗。干燥后浓缩得粗品900mg，直接用于下一步反应。Compound 22i-rac: Compound 22h-rac (2.10 g, 4.65 mmol) was dissolved in 30 mL of dry dichloromethane, cooled in ice water bath, and boron tribromide (2.33 g, 9.29 mmol) was slowly added dropwise and cooled. Stir for 2 hours. The reaction solution was poured slowly into 50 mL of ice water, and 50 mL of dichloromethane was added, and the mixture was separated, and the aqueous layer was extracted with dichloromethane (50 mL × 2), and the organic layer was combined and washed with saturated sodium hydrogen carbonate and brine. After drying, it was concentrated to give a crude material (yield: 900 mg).

化合物22j-rac：将化合物22i-rac(218mg，0.60mmol)溶于20mLDMF中，加入中间体2b(100mg，0.60mmol)和碳酸钾(166mg，1.20mmol)。于85℃加热搅拌3小时，冷却至室温，加入60mL水和50mL乙酸乙酯，分液，水层用乙酸乙酯萃取(50mL×2)，合并有机层，用饱和氯化钠洗，干燥后浓缩得粗品300mg，直接用于下一步反应。Compound 22j-rac: Compound 22i-rac (218 mg, 0.60 mmol) was dissolved in 20 mL of DMF, and Intermediate 2b (100 mg, 0.60 mmol) and potassium carbonate (166 mg, 1.20 mmol). The mixture was heated and stirred at 85 ° C for 3 hours, cooled to room temperature, and then added with 60 mL of water and 50 mL of ethyl acetate. The crude product was concentrated to 300 mg and used directly for the next reaction.

化合物I-22-rac：将化合物22j-rac(200mg，0.46mmol)溶于10mL四氢呋喃和5mL水中，加入氢氧化钠(74mg，1.85mmol)，室温搅拌16小时。用1N盐酸将pH调至1到2，加入20mL二氯甲烷，分液，水层用二氯甲烷萃取(30mL×2)，合并有机层，用饱和氯化钠洗一次，干燥后浓缩，粗品经制备板纯化得产物34mg。收率：18.3％.1H NMR(400MHz，CDCl3)δ：8.47(s，1H)，7.77(s，1H)，7.04(s，1H)，6.74-6.76(d，J＝6.0Hz，1H)，5.05(s，1H)，3.91-4.10(m，4H)，3.34-3.40(m，1H)，3.11-3.15(d，J＝15.6Hz，1H)，2.21-2.31(m，2H)，1.72-1.82(m，2H)，1.25(s，1H)，0.94-0.95(d，J＝6.4Hz，3H)，0.82-0.84(d，J＝6.8Hz，3H).ESI-MS m/z 404.1(M+H)+.Compound I-22-rac: Compound 22j-rac (200 mg, 0.46 mmol) was dissolved in 10 mL of THF and 5 mL of water, and sodium hydroxide (74 mg, 1.85 mmol) was added and stirred at room temperature for 16 hours. The pH was adjusted to 1 to 2 with 1N hydrochloric acid, 20 mL of dichloromethane was added, the mixture was separated, and the aqueous layer was extracted with dichloromethane (30 mL×2). The organic layer was combined, washed once with saturated sodium chloride, dried and concentrated. The product was purified by preparative plate to give 34 mg. Yield: 18.3%. 1 H NMR (400 MHz, CDCl 3 ) δ: 8.47 (s, 1H), 7.77 (s, 1H), 7.04 (s, 1H), 6.74-6.76 (d, J = 6.0 Hz, 1H) ), 5.05 (s, 1H), 3.91-4.10 (m, 4H), 3.34 - 3.40 (m, 1H), 3.11-3.15 (d, J = 15.6 Hz, 1H), 2.21-2.31 (m, 2H), 1.72-1.82 (m, 2H), 1.25 (s, 1H), 0.94-0.95 (d, J = 6.4 Hz, 3H), 0.82-0.84 (d, J = 6.8 Hz, 3H). ESI-MS m/z 404.1 (M+H) + .

实施例24.制备10-氯-6-异丙基-9-((1R，3R)-3-甲氧基环丁氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-23-rac)Example 24. Preparation of 10-chloro-6-isopropyl-9-((1R,3R)-3-methoxycyclobutoxy)-2-oxo-6,7-dihydro-2H-pyridine And [2,1-a]isoquinoline-3-carboxylic acid (I-23-rac)

Figure PCTCN2017094946-appb-000036
Figure PCTCN2017094946-appb-000036

化合物23a-rac：将化合物22i-rac(80mg，0.25mmol)溶于5mLDMF中，加入中间体5c(90mg，0.50mmol)和碳酸钾(166mg，1.20mmol)。于85℃加热搅拌3小时，冷却至室温，加入60mL水和50mL乙酸乙酯，分液，水层用乙酸乙酯萃取(50mL×2)，合并有机层，用饱和氯化钠洗，干燥后浓缩得粗品100mg，直接用于下一步反应。Compound 23a-rac: Compound 22i-rac (80 mg, 0.25 mmol) was dissolved in 5 mL of DMF, and Intermediate 5c (90 mg, 0.50 mmol) and potassium carbonate (166 mg, 1.20 mmol). The mixture was heated and stirred at 85 ° C for 3 hours, cooled to room temperature, and then added with 60 mL of water and 50 mL of ethyl acetate. The crude product was concentrated to 100 mg and used directly for the next reaction.

化合物1-23-rac：将化合物23a-rac(100mg，0.22mmol)溶于10mL四氢呋喃和5mL水中，加入氢氧化钠(40mg，1.00mmol)，室温搅拌16小时。用1N盐酸将pH调至1到2，加入20mL二氯甲烷，分液，水层用二氯甲烷萃取(30mL×2)，合并有机层，用饱和氯化钠洗一次，干燥后浓缩，粗品经制备板纯化得产物17mg。收率：18.5％.1H NMR(400MHz，CDCl3)δ：8.47(s，1H)，7.75(s，1H)，7.04(s，1H)，6.89(s，1H)，4.95(m，1H)，4.15(m，1H)，3.90(m，1H)，3.33(m，4H，overlap)，3.13(m，1H)，2.50(m，4H)，1.77(m，1H)，0.95(d，J＝6.8Hz，3H)，0.84(d，J＝6.8Hz，3H)；ESI-MS m/z 418.2(M+H)+. Compound 1-2-3-rac: Compound 23a-rac (100 mg, 0.22 mmol) was dissolved in 10 mL of THF and 5 mL of water, and sodium hydroxide (40 mg, 1.00 mmol) was added and stirred at room temperature for 16 hours. The pH was adjusted to 1 to 2 with 1N hydrochloric acid, 20 mL of dichloromethane was added, the mixture was separated, and the aqueous layer was extracted with dichloromethane (30 mL×2). The organic layer was combined, washed once with saturated sodium chloride, dried and concentrated. The product was purified by preparative plate to give 17 mg. Yield: 18.5%. 1 H NMR (400MHz, CDCl 3 ) δ: 8.47 (s, 1H), 7.75 (s, 1H), 7.04 (s, 1H), 6.89 (s, 1H), 4.95 (m, 1H) ), 4.15 (m, 1H), 3.90 (m, 1H), 3.33 (m, 4H, overlap), 3.13 (m, 1H), 2.50 (m, 4H), 1.77 (m, 1H), 0.95 (d, J = 6.8 Hz, 3H), 0.84 (d, J = 6.8 Hz, 3H); ESI-MS m/z 418.2 (M+H) + .

实施例25.制备10-氯-6-异丙基-9-((1-(甲氧基甲基)环丙基)甲氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-24-rac)Example 25. Preparation of 10-chloro-6-isopropyl-9-((1-(methoxymethyl)cyclopropyl)methoxy)-2-oxo-6,7-dihydro-2H -pyrido[2,1-a]isoquinoline-3-carboxylic acid (I-24-rac)

Figure PCTCN2017094946-appb-000037
Figure PCTCN2017094946-appb-000037

化合物24a-rac：将化合物22i-rac(80mg，0.25mmol)溶于5mLDMF中，加入中间体6c(90mg，0.50mmol)和碳酸钾(138mg，1.00mmol)。于85℃加热搅拌3小时，冷却至室温，加入60mL水和50mL乙酸乙酯，分液，水层用乙酸乙酯萃取(50mL×2)，合并有机层，用饱和氯化钠洗，干燥后浓缩得粗品110mg，直接用于下一步反应。Compound 24a-rac: Compound 22i-rac (80 mg, 0.25 mmol) was dissolved in 5 mL DMF, and Intermediate 6c (90 mg, 0.50 mmol) and potassium carbonate (138 mg, 1.00 mmol). The mixture was heated and stirred at 85 ° C for 3 hours, cooled to room temperature, and then added with 60 mL of water and 50 mL of ethyl acetate. The crude product was concentrated to 110 mg and used directly for the next reaction.

化合物I-24-rac：将化合物24a-rac(110mg，0.24mmol)溶于10mL四氢呋喃和5mL水中，加入氢氧化钠(40mg，1.00mmol)，室温搅拌16小时。用1N盐酸将pH调至1到2，加入20mL二氯甲烷，分液，水层用二氯甲烷萃取(30mL×2)，合并有机层，用饱和氯化钠洗一次，干燥后浓缩，粗品经制备板纯化得产物23mg。收率：22.2％.1H NMR(400MHz，CDCl3)δ：8.45(s，1H)，7.75(s，1H)，7.03(s，1H)，6.81(s，1H)，4.04(m，2H)，3.90(m，1H)，3.42(m.2H)，3.37(s，3H)，3.33(m，1H)，3.13-3.09(m，1H)，1.77(m，1H)，0.95(d，J＝6.8Hz，3H)，0.82(d，J＝6.4Hz，3H)，0.72(m，2H)，0.66(m，2H)；ESI-MS m/z 432.1(M+H)+.Compound I-24-rac: Compound 24a-rac (110 mg, 0.24 mmol) was dissolved in 10 mL of tetrahydrofuran and 5 mL water, and sodium hydroxide (40 mg, 1.00 mmol) was added and stirred at room temperature for 16 hours. The pH was adjusted to 1 to 2 with 1N hydrochloric acid, 20 mL of dichloromethane was added, the mixture was separated, and the aqueous layer was extracted with dichloromethane (30 mL×2). The organic layer was combined, washed once with saturated sodium chloride, dried and concentrated. The product was purified by preparative plate to give 23 mg. Yield: 22.2%. 1 H NMR (400MHz, CDCl 3 ) δ: 8.45 (s, 1H), 7.75 (s, 1H), 7.03 (s, 1H), 6.81 (s, 1H), 4.04 (m, 2H) ), 3.90 (m, 1H), 3.42 (m. 2H), 3.37 (s, 3H), 3.33 (m, 1H), 3.13 - 3.09 (m, 1H), 1.77 (m, 1H), 0.95 (d, J = 6.8 Hz, 3H), 0.82 (d, J = 6.4 Hz, 3H), 0.72 (m, 2H), 0.66 (m, 2H); ESI-MS m/z 432.1 (M+H) + .

实施例26.制备10-氯-6-异丙基-9-((3-(甲氧基甲基)氧杂环丁烷-3-基)甲氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-α]异喹啉-3-羧酸(I-25-rac)Example 26. Preparation of 10-chloro-6-isopropyl-9-((3-(methoxymethyl)oxetan-3-yl)methoxy)-2-oxo-6, 7-Dihydro-2H-pyrido[2,1-α]isoquinoline-3-carboxylic acid (I-25-rac)

Figure PCTCN2017094946-appb-000038
Figure PCTCN2017094946-appb-000038

化合物25a-rac：将化合物22i-rac(80mg，0.25mmol)溶于5mLDMF中，加入中间体7e(105mg，0.50mmol)和碳酸钾(136mg，1.00mmol)。于85℃加热搅拌3小时，冷却至室温，加入60mL水和50mL乙酸乙酯，分液，水层用乙酸乙酯萃取(50mL×2)，合并有机层，用饱和氯化钠洗，干燥后浓缩得粗品105mg，直接用于下一步反应。Compound 25a-rac: Compound 22i-rac (80 mg, 0.25 mmol) was dissolved in 5 mL of DMF, and Intermediate 7e (105 mg, 0.50 mmol) and potassium carbonate (136 mg, 1.00 mmol). The mixture was heated and stirred at 85 ° C for 3 hours, cooled to room temperature, and then added with 60 mL of water and 50 mL of ethyl acetate. The crude product was concentrated to 105 mg, which was used directly for the next reaction.

化合物I-25-rac：将化合物25a-rac(105mg，0.22mmol)溶于10mL四氢呋喃和5mL水中，加入氢氧化钠(40mg，1.00mmol)，室温搅拌16小时。用1N盐酸将pH调至1到2，加入20mL二氯甲烷，分液，水层用二氯甲烷萃取(30mL×2)，合并有机层，用饱和氯化钠洗一次，干燥后浓缩，粗品经制备板纯化得产物21mg。收率：21.3％.1H NMR(400MHz，CDCl3)δ：8.48(s，1H)，7.76(s，1H)，7.04(s，1H)，6.89(s，1H)，4.62(m，2H)，4.55(m，2H)，4.30(q，J＝8.8Hz，2H)，3.95(m，1H)，3.79(m，2H)，3.65(s，3H)，3.37(m，1H)，3.13(m，1H)，1.70(m，1H)，0.95(d，J＝6.4Hz，3H)，0.82(d，J＝6.8Hz，3H).ESI-MS m/z 448.1(M+H)+.Compound I-25-rac: Compound 25a-rac (105 mg, 0.22 mmol) was dissolved in 10 mL of tetrahydrofuran and 5 mL water, and sodium hydroxide (40 mg, 1.00 mmol) was added and stirred at room temperature for 16 hours. The pH was adjusted to 1 to 2 with 1N hydrochloric acid, 20 mL of dichloromethane was added, the mixture was separated, and the aqueous layer was extracted with dichloromethane (30 mL×2). The organic layer was combined, washed once with saturated sodium chloride, dried and concentrated. The product was purified by preparative plate to give 21 mg. Yield: 21.3%. 1 H NMR (400MHz, CDCl 3 ) δ: 8.48 (s, 1H), 7.76 (s, 1H), 7.04 (s, 1H), 6.89 (s, 1H), 4.62 (m, 2H) ), 4.55 (m, 2H), 4.30 (q, J = 8.8 Hz, 2H), 3.95 (m, 1H), 3.79 (m, 2H), 3.65 (s, 3H), 3.37 (m, 1H), 3.13 (m, 1H), 1.70 (m, 1H), 0.95 (d, J = 6.4 Hz, 3H), 0.82 (d, J = 6.8 Hz, 3H). ESI-MS m/z 448.1 (M+H) + .

实施例27.制备10-氯-6-异丙基-9-((3-甲氧基环丁基)甲氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-26-rac)Example 27. Preparation of 10-chloro-6-isopropyl-9-((3-methoxycyclobutyl)methoxy)-2-oxo-6,7-dihydro-2H-pyridine and [ 2,1-a]isoquinoline-3-carboxylic acid (I-26-rac)

Figure PCTCN2017094946-appb-000039
Figure PCTCN2017094946-appb-000039

化合物26a-rac：将化合物22i-rac(186mg，0.52mmol)溶于20mLDMF中，加入中间体8d(100mg，0.52mmol)和碳酸钾(142mg，1.04mmol)。于85℃加热搅拌3小时，冷却至室温，加入60mL水，然后用乙酸乙酯萃取(50mL×3)，合并有机层，用饱和氯化钠洗一次，干燥后浓缩得粗品200mg，直接用于下一步反应。 Compound 26a-rac: Compound 22i-rac (186 mg, 0.52 mmol) was dissolved in 20 mL of DMF, and Intermediate 8d (100mg, 0.52mmol) and potassium carbonate (142mg, 1.04mmol). The mixture was heated and stirred at 85 ° C for 3 hours, cooled to room temperature, then added with 60 mL of water, and then extracted with ethyl acetate (50 mL × 3). The organic layer was combined, washed with saturated sodium chloride, dried and concentrated The next step is to react.

化合物I-26-rac：将化合物26a-rac(200mg，0.44mmol)溶于10mLTHF和5mL水中，加入氢氧化钠(70mg，1.74mmol)，室温搅拌16小时。浓缩THF，用1N盐酸将pH调至1到2，用DCM萃取(30mL×3)，合并有机层，用饱和氯化钠洗一次，干燥后浓缩。粗产品经制备板纯化得产物12mg。收率：6.32％。1H NMR(400MHz，CDCl3)δ8.47(s，1H)，7.74(s，1H)，7.02(s，1H)，6.79(s，1H)，4.06-4.10(m，2H)，3.92-3.95(m，1H)，3.83-3.87(m，1H)，3.34-3.39(m，1H)，3.25(s，1H)，3.10-3.14(d，J＝16.4Hz，1H)，2.37-2.52(m，3H)，1.80-1.89(m，2H)，1.21(s，1H)，0.94-0.95(d，J＝6.4Hz，3H)，0.81-0.83(d，J＝6.8Hz，3H).ESI-MS m/z 432.2(M+H)+。Compound I-26-rac: Compound 26a-rac (200 mg, 0.44 mmol) was dissolved in 10 mL THF and 5 mL water, and sodium hydroxide (70 mg, 1.74 mmol) was added and stirred at room temperature for 16 hours. The THF was concentrated, the pH was adjusted to 1 to 2 with 1N hydrochloric acid, and the mixture was evaporated. The crude product was purified on a preparative plate to afford 12 mg. Yield: 6.32%. 1 H NMR (400MHz, CDCl 3 ) δ8.47 (s, 1H), 7.74 (s, 1H), 7.02 (s, 1H), 6.79 (s, 1H), 4.06-4.10 (m, 2H), 3.92- 3.95 (m, 1H), 3.83-3.87 (m, 1H), 3.34-3.39 (m, 1H), 3.25 (s, 1H), 3.10-3.14 (d, J = 16.4 Hz, 1H), 2.37-2.52 ( m, 3H), 1.80-1.89 (m, 2H), 1.21 (s, 1H), 0.94-0.95 (d, J = 6.4 Hz, 3H), 0.81 - 0.83 (d, J = 6.8 Hz, 3H). - MS m/z 432.2 (M+H) + .

实施例28.制备6-(叔丁基)-10-甲氧基-2-氧代-9-(((R)-四氢呋喃-3-基)氧基)-6，7-二氢-2H-吡啶并[2，1-α]异喹啉-3-羧酸(I-28-rac)Example 28. Preparation of 6-(tert-butyl)-10-methoxy-2-oxo-9-(((R)-tetrahydrofuran-3-yl)oxy)-6,7-dihydro-2H -pyrido[2,1-α]isoquinoline-3-carboxylic acid (I-28-rac)

Figure PCTCN2017094946-appb-000040
Figure PCTCN2017094946-appb-000040

化合物28a：将化合物2(4.40g，15mmol)溶于四氢呋喃(50mL)中，加入Pd2(dba)3(275mg，0.3mmol)，Xantphos(347mg，0.6mmol)和叔丁醇钠(2.30g，24mmol)。氮气置换3次，加入频哪酮(3.00g，15mmol)，于60℃加热搅拌6小时。加入硅胶拌样，柱纯化得产物3.30g。收率：70.7％。Compound 28a: Compound 2 (4.40 g, 15 mmol) was dissolved in tetrahydrofuran (50 mL), Pd 2 (dba) 3 (275 mg, 0.3 mmol), Xantphos (347 mg, 0.6 mmol) and sodium tert-butoxide (2.30 g, 24mmol). The mixture was replaced with nitrogen three times, and pinacolone (3.00 g, 15 mmol) was added, and the mixture was stirred under heating at 60 ° C for 6 hours. The mixture was added to a silica gel and the column was purified to give 3.30 g. Yield: 70.7%.

化合物28b-rac：将化合物28a(3.30g，10.6mmol)溶于50mL甲醇中，加入醋酸铵(8.14g，0.11mol)和氰基硼氢化钠(995mg，15.8mmol)，温搅拌16小时。浓缩甲醇，加入NaOH水溶液(1.40g氢氧化钠溶于60mL水)和50mLDCM，搅拌20分钟。分液，水层用DCM萃取(50mL×2)，合并有机层，用饱和氯化钠洗一次，无水硫酸钠干燥，浓缩得粗产品3.40g，直接用于下一步反应。Compound 28b-rac: Compound 28a (3.30 g, 10.6 mmol) was dissolved in 50 mL of MeOH. EtOAc (EtOAc, EtOAc, EtOAc. Methanol was concentrated, and aqueous NaOH (1.40 g of sodium hydroxide dissolved in 60 mL of water) and 50 m LDCM were added and stirred for 20 minutes. The organic layer was combined and washed with saturated sodium chloride, dried over anhydrous sodium sulfate and evaporated.

化合物28c-rac：将化合物28b-rac(3.40g，10.9mmol)溶于50mL二氧六环中，加入甲酸(2.51g，54.5mmol)，加热回流3小时。冷却至室温，加入50mL饱和碳酸氢钠水溶液，然后用乙酸乙酯萃取(50mL×3)，合并有机层，用饱和氯化钠洗一次，无水硫酸钠干燥，浓缩得粗品3.40g，直接用于下一步反应。Compound 28c-rac: Compound 28b-rac (3.40 g, 10.9 mmol) was dissolved in 50 mL of dioxane, and toluene (2.51 g, 54.5 mmol). After cooling to room temperature, 50 mL of a saturated aqueous solution of sodium hydrogencarbonate was added, and then ethyl acetate (50 mL × 3), and the organic layer was combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate and evaporated. In the next step.

化合物28d-rac：将化合物28c-rac(3.40g，10mmol)溶于50mL乙腈中，加入三氯氧磷(1.85g，12mmol)，室温搅拌16小时。将反应液缓慢倒入50mL水中，浓缩乙腈，用氨水将pH调至8-9，然后用DCM萃取(50mL×3)，合并有机层，用饱和氯化钠洗，无水硫酸钠干燥，浓缩得粗品3.20g，直接用于下一步反应。Compound 28d-rac: Compound 28c-rac (3.40 g, 10 mmol) was dissolved in 50 mL of acetonitrile, and phosphorus oxychloride (1.85 g, 12 mmol) was added and stirred at room temperature for 16 hours. The reaction mixture was poured into 50 mL of water, and the mixture was concentrated with EtOAc (EtOAc). A crude product of 3.20 g was obtained and used directly for the next reaction.

化合物28e-rac：将化合物28d-rac(3.20g，9.9mmol)溶于40mL乙醇和5mL水中，加入2-乙氧亚甲基乙酰乙酸乙酯(5.50g，29.7mmol)，于80℃加热搅拌4小时。浓缩乙醇和水得粗品6.40g，直接用于下一步反应。Compound 28e-rac: Compound 28d-rac (3.20 g, 9.9 mmol) was dissolved in 40 mL of ethanol and 5 mL of water, and ethyl 2-acetoxymethylacetate (5.50 g, 29.7 mmol) was added and stirred at 80 ° C. 4 hours. The ethanol and water were concentrated to give a crude product, 6.40 g, which was directly used for the next reaction.

化合物28f-rac：将化合物28e-rac(6.40g，13.8mmol)溶于40mL乙二醇二甲醚中，加入四氯苯醌(3.40g，13.8mmol)，于55℃加热搅拌3小时。加入硅胶拌样，柱层析纯化得产品3.20g。收率：50.2％。Compound 28f-rac: Compound 28e-rac (6.40 g, 13.8 mmol) was dissolved in 40 mL of ethylene glycol dimethyl ether, and tetrachlorophenylhydrazine (3.40 g, 13.8 mmol) was added, and the mixture was stirred under heating at 55 ° C for 3 hours. The mixture was added to a silica gel and purified by column chromatography to give a product 3.20 g. Yield: 50.2%.

化合物28g-rac：将化合物28f-rac(3.2g，6.9mmol)溶于50mL甲醇中，加入200mg Pd/C。氢气置换4次，室温搅 拌16小时。垫硅藻土抽滤，浓缩得粗产品2.40g。Compound 28g-rac: Compound 28f-rac (3.2 g, 6.9 mmol) was dissolved in 50 mL of methanol, and 200 mg of Pd/C was added. Hydrogen replacement 4 times, stirring at room temperature Mix for 16 hours. The padded diatomaceous earth was suction filtered and concentrated to yield 2.40 g.

化合物28h-rac：将化合物28g-rac(223mg，0.60mmol)溶于20mL DMF中，加入中间体2b(100mg，0.60mmol)和碳酸钾(166mg，1.20mmol)。于85℃加热搅拌3小时，冷却至室温，加入60mL水和50mL乙酸乙酯，分液，水层用50mL乙酸乙酯萃取，合并有机层，用饱和氯化钠洗一次，浓缩得粗品300mg，直接用于下一步反应。Compound 28h-rac: Compound 28 g- rac (223 mg, 0.60 mmol) was dissolved in 20 mL DMF, and Intermediate 2b (100 mg, 0.60 mmol) and potassium carbonate (166 mg, 1.20 mmol). The mixture was heated and stirred at 85 ° C for 3 hours, cooled to room temperature, then added with 60 mL of water and 50 mL of ethyl acetate. The mixture was separated and evaporated. Used directly in the next step.

化合物I-28-rac：将化合物28h-rac(300mg，0.68mmol)溶于10mL THF和5mL水中，加入氢氧化钠(109mg，2.72mmol)，室温搅拌16小时。用1N盐酸将pH调至1到2，用DCM萃取(30mL×3)，合并有机层，用饱和氯化钠洗一次，干燥后浓缩，粗产品经制备板纯化得产物100mg。收率：35.5％。1H NMR(400MHz，CDCl3)δ：8.47(s，1H)，7.16(s，1H)，7.07(s，1H)，6.65-6.66(d，J＝4.8Hz，1H)，5.04(s，1H)，4.01-4.06(m，4H)，3.91(s，3H)，3.39-3.45(m，1H)，3.14-3.18(d，J＝16.8Hz，1H)，2.21-2.26(m，2H)，1.25(s，1H)，0.81(s，9H).ESI-MS m/z 414.2(M+H)+.Compound I-28-rac: Compound 28h-rac (300 mg, 0.68 mmol) was dissolved in 10 mL THF and 5 mL water, and sodium hydroxide (109 mg, 2.72 mmol) was added and stirred at room temperature for 16 hours. The pH was adjusted to 1 to 2 with 1N hydrochloric acid and extracted with DCM (30 mL×3). The organic layer was combined, washed once with saturated sodium chloride, dried and concentrated. Yield: 35.5%. 1 H NMR (400MHz, CDCl 3 ) δ: 8.47 (s, 1H), 7.16 (s, 1H), 7.07 (s, 1H), 6.65-6.66 (d, J = 4.8Hz, 1H), 5.04 (s, 1H), 4.01-4.06 (m, 4H), 3.91 (s, 3H), 3.39-3.45 (m, 1H), 3.14 - 3.18 (d, J = 16.8 Hz, 1H), 2.21-2.26 (m, 2H) , 1.25 (s, 1H), 0.81 (s, 9H). ESI-MS m/z 414.2 (M+H) + .

实施例29.制备6-(叔丁基)-10-甲氧基-9-((1R，3R)-3-甲氧基环丁氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-α]异喹啉-3-羧酸(I-29-rac)Example 29. Preparation of 6-(tert-butyl)-10-methoxy-9-((1R,3R)-3-methoxycyclobutoxy)-2-oxo-6,7-dihydro -2H-pyrido[2,1-α]isoquinoline-3-carboxylic acid (I-29-rac)

Figure PCTCN2017094946-appb-000041
Figure PCTCN2017094946-appb-000041

化合物29a-rac：将化合物28g-rac(65mg，0.18mmol)，化合物5c(47mg，0.26mmol)和碳酸钾(50mg，0.36mmol)加入到5mL DMF中，置换氮气3次，于90℃加热搅拌反应3小时。反应完毕后加水稀释，用乙酸乙酯萃取(20mL×3)，合并有机相，无水硫酸钠干燥，浓缩得粗品75mg。Compound 29a-rac: Compound 28g-rac (65mg, 0.18mmol), compound 5c (47mg, 0.26mmol) and potassium carbonate (50mg, 0.36mmol) were added to 5mL DMF, replaced with nitrogen three times, heated and stirred at 90 °C Reaction for 3 hours. After completion of the reaction, the mixture was diluted with EtOAc (EtOAc m.

化合物I-29-rac：将化合物29a-rac(75mg，0.16mmol)溶于4mL四氢呋喃中，加入氢氧化钠45mg和水1mL，于50℃搅拌反应3小时。反应完毕后用1N盐酸调pH至2-3左右，二氯甲烷萃取(20mL×3)，合并有机相，无水硫酸钠干燥，浓缩后薄层层析得产品10mg。收率：14.6％。1H-NMR(CDCl3，400MHz)δ：8.45(s，1H)，7.14(s，1H)，7.05(s，1H)，6.51(s，1H)，4.94(m，1H)，4.16(m，1H)，4.01(m，1H)，3.92(s，3H)，3.37-3.43(m，1H)，3.29(s，3H)，3.16(m，1H)，2.48-2.56(m，3H)，0.82(s，9H)；ESI-MS m/z 428.2(M+H)+ Compound I-29-rac: Compound 29a-rac (75 mg, 0.16 mmol) was dissolved in 4 mL of tetrahydrofuran, and sodium hydroxide 45 mg and water 1 mL were added, and the reaction was stirred at 50 ° C for 3 hours. After the completion of the reaction, the pH was adjusted to about 2-3 with 1N hydrochloric acid, and extracted with dichloromethane (20 mL×3). The organic phase was combined and dried over anhydrous sodium sulfate. Yield: 14.6%. 1 H-NMR (CDCl 3 , 400 MHz) δ: 8.45 (s, 1H), 7.14 (s, 1H), 7.05 (s, 1H), 6.51 (s, 1H), 4.94 (m, 1H), 4.16 (m) , 1H), 4.01 (m, 1H), 3.92 (s, 3H), 3.37-3.43 (m, 1H), 3.29 (s, 3H), 3.16 (m, 1H), 2.48-2.56 (m, 3H), 0.82 (s, 9H); ESI-MS m/z 428.2 (M+H) +

实施例30.制备6-(叔丁基)-10-甲氧基-9-((1-(甲氧基甲基)环丙基)甲氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-α]异喹啉-3-羧酸(I-30-rac)Example 30. Preparation of 6-(tert-butyl)-10-methoxy-9-((1-(methoxymethyl)cyclopropyl)methoxy)-2-oxo-6,7- Dihydro-2H-pyrido[2,1-α]isoquinoline-3-carboxylic acid (I-30-rac)

Figure PCTCN2017094946-appb-000042
Figure PCTCN2017094946-appb-000042

化合物30a-rac：将化合物28g-rac(85mg，0.23mmol)，化合物6c(89mg，0.46mmol)和碳酸钾(63mg，0.46mmol)加入到5mL DMF中，置换氮气3次，于90℃加热搅拌反应3小时。反应完毕后加水稀释，用乙酸乙酯萃取(20mL×3)，合并有机相，无水硫酸钠干燥，浓缩得粗品97mg。Compound 30a-rac: Compound 28g-rac (85mg, 0.23mmol), compound 6c (89mg, 0.46mmol) and potassium carbonate (63mg, 0.46mmol) were added to 5mL DMF, replaced with nitrogen three times, heated and stirred at 90 °C Reaction for 3 hours. After completion of the reaction, the mixture was diluted with EtOAc (EtOAc m.

化合物I-30-rac：将化合物29a-rac(97mg，0.21mmol)溶于4mL四氢呋喃中，加入氢氧化钠45mg和水1mL，于50℃搅拌反应3小时。反应完毕后用1N盐酸调pH至2-3左右，二氯甲烷萃取(20mL×3)，合并有机相，无水硫酸钠干燥，浓缩后薄层层析得产品26mg。收率：28.1％。1H-NMR(CDCl3，400MHz)δ：8.45(s，1H)，7.13(s，1H)，7.05(s，1H)，6.74(s，1H)，4.00(m，3H)，3.91(s，3H)，3.41(s，2H)，3.36(s，3H)，3.15(m，1H)，2.92(m，1H)，0.82(s，9H)，0.69-0.65(m，4H)；ESI-MS m/z 442.2(M+H)+。Compound I-30-rac: Compound 29a-rac (97 mg, 0.21 mmol) was dissolved in 4 mL of tetrahydrofuran, and sodium hydroxide (45 mg) and water (1 mL) were added, and the mixture was stirred at 50 ° C for 3 hours. After completion of the reaction, the pH was adjusted to about 2-3 with 1N hydrochloric acid and extracted with dichloromethane (20 mL×3). The organic phase was combined and dried over anhydrous sodium sulfate. Yield: 28.1%. 1 H-NMR (CDCl 3 , 400 MHz) δ: 8.45 (s, 1H), 7.13 (s, 1H), 7.05 (s, 1H), 6.74 (s, 1H), 4.00 (m, 3H), 3.91 (s) , 3H), 3.41 (s, 2H), 3.36 (s, 3H), 3.15 (m, 1H), 2.92 (m, 1H), 0.82 (s, 9H), 0.69-0.65 (m, 4H); MS m/z 442.2 (M + H) + .

实施例31.制备6-(叔丁基)-10-甲氧基-9-((3-(甲氧基甲基)氧杂环丁烷-3-基)甲氧基)-2-氧代-6，7-二氢-2H-吡啶并 [2，1-α]异喹啉-3-羧酸(I-31-rac)Example 31. Preparation of 6-(tert-butyl)-10-methoxy-9-((3-(methoxymethyl)oxetan-3-yl)methoxy)-2-oxo -6,7-dihydro-2H-pyridine [2,1-α]isoquinoline-3-carboxylic acid (I-31-rac)

Figure PCTCN2017094946-appb-000043
Figure PCTCN2017094946-appb-000043

化合物31a-rac：将化合物28g-rac(65mg，0.18mmol)，化合物7e(55mg，0.26mmol)和碳酸钾(50mg，0.36mmol)加入到5mL DMF中，置换氮气3次，于90℃加热搅拌反应3小时。反应完毕后加水稀释，用乙酸乙酯萃取(20mL×3)，合并有机相，无水硫酸钠干燥，浓缩得粗品87mg。Compound 31a-rac: Compound 28g-rac (65mg, 0.18mmol), compound 7e (55mg, 0.26mmol) and potassium carbonate (50mg, 0.36mmol) were added to 5mL DMF, replaced with nitrogen three times, heated and stirred at 90 °C Reaction for 3 hours. After completion of the reaction, the mixture was diluted with EtOAc (EtOAc m.

化合物I-31-rac：将化合物31a-rac(87mg，0.18mmol)溶于4mL四氢呋喃中，加入氢氧化钠45mg和水1mL，于50℃搅拌反应3小时。反应完毕后用1N盐酸调pH至2-3左右，二氯甲烷萃取(20mL×3)，合并有机相，无水硫酸钠干燥，浓缩后薄层层析得产品10mg。收率：12.1％。1H-NMR(CDCl3，400MHz)δ：8.46(s，1H)，7.15(s，1H)，7.07(s，1H)，6.79(s，1H)，4.61(m，4H)，4.27(m，2H)，4.01(m，1H)，3.89(s，3H)，3.78(m，2H)，3.41(s，3H)，3.33(m，1H)，3.16(m，1H)，0.83(s，9H)；ESI-MS m/z 458.2(M+H)+。Compound I-31-rac: Compound 31a-rac (87 mg, 0.18 mmol) was dissolved in 4 mL of tetrahydrofuran, and sodium hydroxide 45 mg and water 1 mL were added, and the reaction was stirred at 50 ° C for 3 hours. After the completion of the reaction, the pH was adjusted to about 2-3 with 1N hydrochloric acid, and extracted with dichloromethane (20 mL×3). The organic phase was combined and dried over anhydrous sodium sulfate. Yield: 12.1%. 1 H-NMR (CDCl 3 , 400 MHz) δ: 8.46 (s, 1H), 7.15 (s, 1H), 7.07 (s, 1H), 6.79 (s, 1H), 4.61 (m, 4H), 4.27 (m) , 2H), 4.01 (m, 1H), 3.89 (s, 3H), 3.78 (m, 2H), 3.41 (s, 3H), 3.33 (m, 1H), 3.16 (m, 1H), 0.83 (s, 9H); ESI-MS m/z 458.2 (M+H) + .

实施例32.制备6-(叔丁基)-10-甲氧基-9-((3-甲氧基环丁基)甲氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-32-rac)Example 32. Preparation of 6-(tert-butyl)-10-methoxy-9-((3-methoxycyclobutyl)methoxy)-2-oxo-6,7-dihydro-2H -pyrido[2,1-a]isoquinoline-3-carboxylic acid (I-32-rac)

Figure PCTCN2017094946-appb-000044
Figure PCTCN2017094946-appb-000044

化合物32a-rac：将化合物28g-rac(191mg，0.52mmol)，化合物8d(100mg，0.52mmol)和碳酸钾(142mg，1.03mmol)加入到10mL DMF中，置换氮气3次，于90℃加热搅拌反应5小时。反应完毕后加水稀释，用乙酸乙酯萃取(50mL×3)，合并有机相，无水硫酸钠干燥，浓缩后得粗产品200mg。Compound 32a-rac: Compound 28g-rac (191mg, 0.52mmol), compound 8d (100mg, 0.52mmol) and potassium carbonate (142mg, 1.03mmol) were added to 10mL DMF, replaced with nitrogen 3 times, heated and stirred at 90 °C Reaction for 5 hours. After the completion of the reaction, the mixture was diluted with water and evaporated with ethyl acetate (50 mL×3).

化合物I-32-rac：将化合物32a-rac(200mg，0.43mmo)溶于10mL四氢呋喃中，加入氢氧化钠102mg和水5mL，于35℃搅拌反应2小时。反应完毕后用1N盐酸调pH至2-3左右，二氯甲烷萃取(20mL×3)，合并有机相，无水硫酸钠干燥，浓缩后薄层层析得产品70mg。收率：36.8％。1H-NMR(CDCl3，400MHz)δ：8.48(s，1H)，7.15(s，1H)，7.07(s，1H)，6.71(s，1H)，4.03-4.08(m，3H)，3.92(s，3H)，3.40-3.46(m，1H)，3.15-3.19(d，J＝15.2Hz，1H)，2.22-2.55(m，3H)，1.78-1.82(m，2H)，1.26(s，1H)，0.82(s，9H).ESI-MS m/z442.2(M+H)+.Compound I-32-rac: Compound 32a-rac (200 mg, 0.43 mmol) was dissolved in 10 mL of tetrahydrofuran, and sodium hydroxide 102 mg and water 5 mL were added, and the reaction was stirred at 35 ° C for 2 hours. After completion of the reaction, the pH was adjusted to about 2-3 with 1N hydrochloric acid, and extracted with dichloromethane (20 mL×3). The organic phase was combined and dried over anhydrous sodium sulfate. Yield: 36.8%. 1 H-NMR (CDCl 3 , 400 MHz) δ: 8.48 (s, 1H), 7.15 (s, 1H), 7.07 (s, 1H), 6.71 (s, 1H), 4.03-4.08 (m, 3H), 3.92 (s, 3H), 3.40-3.46 (m, 1H), 3.15-3.19 (d, J = 15.2 Hz, 1H), 2.22-2.55 (m, 3H), 1.78-1.82 (m, 2H), 1.26 (s) , 1H), 0.82 (s, 9H). ESI-MS m/z 442.2 (M+H) + .

实施例33.制备10-甲氧基-6-(1-甲氧基-2-甲基丙-2-基)-2-氧代-9-(((R)-四氢呋喃-3-基)氧基)-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-52-rac)Example 33. Preparation of 10-methoxy-6-(1-methoxy-2-methylpropan-2-yl)-2-oxo-9-(((R)-tetrahydrofuran-3-yl) Oxy)-6,7-dihydro-2H-pyrido[2,1-a]isoquinoline-3-carboxylic acid (I-52-rac)

Figure PCTCN2017094946-appb-000045
Figure PCTCN2017094946-appb-000045

Figure PCTCN2017094946-appb-000046
Figure PCTCN2017094946-appb-000046

化合物52b：将化合物52a(10.0g，0.12mol)溶于18mL三氟乙酸中，加入3.48g多聚甲醛，80℃条件下搅拌反应7小时，冷却后加入500mL饱和碳酸氢钠水溶液搅拌反应6小时。然后二氯甲烷萃取(100mL×5)，合并有机相，无水硫酸钠干燥，浓缩拌样，硅胶柱层析(PE∶EA＝5∶1)得产品5.0g。收率：35.8％。Compound 52b: Compound 52a (10.0 g, 0.12 mol) was dissolved in 18 mL of trifluoroacetic acid, 3.48 g of paraformaldehyde was added, and the reaction was stirred at 80 ° C for 7 hours. After cooling, 500 mL of a saturated aqueous solution of sodium hydrogencarbonate was added and the reaction was stirred for 6 hours. . Then, it was extracted with dichloromethane (100 mL × 5), and the organic phase was combined, dried over anhydrous sodium sulfate, and the mixture was concentrated to silica gel column chromatography (PE: EA = 5:1) to obtain 5.0 g of product. Yield: 35.8%.

化合物52c：往化合物52b(5.0g，0.043mol)中加入硫酸二甲酯5.5mL，然后加入20N氢氧化钠水溶液3mL，40℃搅拌反应16小时。TLC监测原料反应完毕，用乙醚萃取(30mL×3)，合并有机相，无水硫酸钠干燥，浓缩蒸干得1.00g粗产品。Compound 52c: To a compound 52b (5.0 g, 0.043 mol), 5.5 mL of dimethyl sulfate was added, and then 3 mL of a 20 N aqueous sodium hydroxide solution was added thereto, and the mixture was stirred at 40 ° C for 16 hours. The reaction of the material was monitored by TLC, eluted with diethyl ether (30mL×3), and the organic phase was combined, dried over anhydrous sodium sulfate and evaporated to dryness

化合物52d：将化合物2(1.00g，3.41mmol)，化合物52c(0.89g，6.82mmol)，醋酸钯(12mg，1.5％mol)，Xphos(2-二环己基磷-2’，4’，6’-三异丙基联苯，49mg，3％mol)和1M LiHMDS(10.2mL，10.2mmol)溶于10mL二氧六环中，氮气保护下置换气3次，70℃条件下加热反应3小时，反应完毕后旋干反应溶剂，硅胶柱层析PE∶EA＝5∶1得产品0.62g。收率：53.4％。Compound 52d: Compound 2 (1.00 g, 3.41 mmol), Compound 52c (0.89 g, 6.82 mmol), palladium acetate (12 mg, 1.5% mol), Xphos (2-dicyclohexylphosphon-2', 4', 6 '-Triisopropyl biphenyl, 49 mg, 3% mol) and 1 M LiHMDS (10.2 mL, 10.2 mmol) were dissolved in 10 mL of dioxane, replaced with nitrogen three times under nitrogen, and heated at 70 ° C for 3 hours. After the reaction was completed, the reaction solvent was spin-dried, and silica gel column chromatography PE:EA=5:1 gave the product 0.62 g. Yield: 53.4%.

化合物52e：将化合物52d(0.62g，1.82mmol)溶于5mL甲醇中，加入醋酸铵(1.40g，18.2mmol)搅拌反应30分钟，加入氰基硼氢化钠(0.23g，3.64mmol)，于60℃搅拌反应16小时。反应完毕后加入2N氢氧化钠溶液50mL，二氯甲烷萃取(30mL×3)，合并有机相，无水硫酸钠干燥，浓缩蒸干得0.63g粗产品。Compound 52e: Compound 52d (0.62 g, 1.82 mmol) was dissolved in 5 mL of MeOH. EtOAc (1. <RTIgt; The reaction was stirred at ° C for 16 hours. After completion of the reaction, 50 mL of a 2N sodium hydroxide solution was added, and dichloromethane (30 mL×3) was evaporated. The organic phase was combined, dried over anhydrous sodium sulfate and evaporated to dryness.

化合物52f-rac：将化合物52e(0.63g，1.84mmol)溶于5mL二氧六环中，加入1.0mL甲酸，0.5mL原甲酸三乙酯，加热回流搅拌反应48小时。反应结束后旋干反应溶剂，二氯甲烷萃取(20mL×3)，合并有机相，无水硫酸钠干燥，浓缩蒸干得0.60g粗产品。Compound 52f-rac: Compound 52e (0.63 g, 1.84 mmol) was dissolved in 5 mL of dioxane, and 1.0 mL of formic acid and 0.5 mL of triethyl orthoformate were added, and the mixture was stirred under reflux for 48 hours. After completion of the reaction, the solvent was evaporated to dryness.

化合物52g-rac：将化合物52f-rac(0.60g，1.62mmol)溶于5mL乙腈中，冰浴条件下缓慢滴加三氯氧磷(0.30mL，3.24mmol)，滴毕油浴60℃反应1小时。反应完毕后旋干反应溶剂，二氯甲烷萃取(20mL×3)，有机相合并，饱和碳酸氢钠溶液洗，无水硫酸钠干燥，浓缩得到0.58g粗产品。Compound 52g-rac: Compound 52f-rac (0.60 g, 1.62 mmol) was dissolved in 5 mL of acetonitrile, and phosphorus oxychloride (0.30 mL, 3.24 mmol) was slowly added dropwise under ice bath, and the reaction was carried out in an oil bath at 60 ° C. hour. After completion of the reaction, the solvent was evaporated to dryness.

化合物52h-rac：将化合物52g-rac(0.58g，1.64mmol)和2-乙氧亚甲基乙酰乙酸乙酯(0.92g，4.92mmol)溶于6mL乙醇中，加入2mL水，加热回流搅拌反应48小时，反应完毕后旋干反应溶剂得1.17g粗产品。 Compound 52h-rac: Compound 52g-rac (0.58g, 1.64mmol) and ethyl 2-ethoxymethyleneacetate (0.92g, 4.92mmol) were dissolved in 6mL of ethanol, 2mL water was added, and the reaction was stirred and heated under reflux. After 48 hours, after completion of the reaction, the reaction solvent was dried to give 1.17 g of crude product.

化合物52i-rac：将化合物52h-rac(1.17g，2.36mmol)和四氯苯醌(0.35g，1.42mmol)溶于15mL乙二醇二甲醚中，加热回流搅拌反应3小时，反应完毕后旋干反应溶剂，硅胶柱层析得产品475mg。收率：40.9％。Compound 52i-rac: Compound 52h-rac (1.17 g, 2.36 mmol) and tetrachlorophenylhydrazine (0.35 g, 1.42 mmol) were dissolved in 15 mL of ethylene glycol dimethyl ether and stirred under reflux for 3 hours. The reaction solvent was spin-dried, and the product was purified by silica gel column chromatography. Yield: 40.9%.

化合物52j-rac：将化合物52i-rac(475mg，0.97mmol)溶于15mL甲醇中，加入钯碳50mg，氢气保护下置换气3次。室温过夜搅拌反应，TLC监测反应完毕，抽滤，滤液蒸干得产品300mg。收率：77.3％。1H-NMR(CDCl3，400MHz)：δ：8.35(d，1H，J＝13.6Hz)，7.11(s，1H)，6.91(s，1H)，6.80(s，1H)，4.37(d，2H，J＝6.4Hz)，3.90(s，3H)，3.86(s，1H)，3.34(s，3H)，3.33-3.28(m，1H)，3.03(s，1H)，2.99-2.94(m，1H)，2.88-2.85(m，1H)，1.38-1.24(m，3H)，0.95(d，3H，J＝4Hz)，0.39(d，3H，J＝6.8Hz)Compound 52j-rac: Compound 52i-rac (475 mg, 0.97 mmol) was dissolved in 15 mL of methanol, 50 mg of palladium carbon was added, and the solvent was replaced with hydrogen for 3 times. The reaction was stirred at room temperature overnight, the reaction was completed by TLC, suction filtered, and the filtrate evaporated to dryness. Yield: 77.3%. 1H-NMR (CDCl 3 , 400MHz): δ: 8.35 (d, 1H, J = 13.6 Hz), 7.11 (s, 1H), 6.91 (s, 1H), 6.80 (s, 1H), 4.37 (d, 2H) , J=6.4Hz), 3.90(s,3H), 3.86(s,1H), 3.34(s,3H),3.33-3.28(m,1H),3.03(s,1H),2.99-2.94(m, 1H), 2.88-2.85 (m, 1H), 1.38-1.24 (m, 3H), 0.95 (d, 3H, J = 4 Hz), 0.39 (d, 3H, J = 6.8 Hz)

化合物52k-rac：将化合物52j-rac(100mg，0.25mmol)，化合物2b(83mg，0.50mmol)和碳酸钾(103mg，0.75mmol)加入到5mL DMF中，置换氮气3次，于90℃加热搅拌反应5小时。反应完毕后加水稀释，用乙酸乙酯萃取(20mL×3)，合并有机相，无水硫酸钠干燥，薄层层析得产品75mg。收率：40.0％。Compound 52k-rac: Compound 52j-rac (100 mg, 0.25 mmol), compound 2b (83 mg, 0.50 mmol) and potassium carbonate (103 mg, 0.75 mmol) were added to 5 mL of DMF, replaced with nitrogen three times, heated and stirred at 90 °C. Reaction for 5 hours. After completion of the reaction, it was diluted with water and extracted with ethyl acetate (20 mL×3). Yield: 40.0%.

化合物I-52-rac：将化合物52k-rac(75mg，0.1mmol)溶于3mL四氢呋喃中，加入氢氧化钠32mg和水1mL，于35℃搅拌反应2小时。反应完毕后用1N盐酸调pH至2-3左右，二氯甲烷萃取(20mL×3)，合并有机相，无水硫酸钠干燥，浓缩后薄层层析得产品30mg。收率：67.5％。1H-NMR(CDCl3，400MHz)δ：8.58(s，1H)，7.15(s，1H)，7.06(s，1H)，6.66(s，1H)，5.05-5.03(m，1H)，4.52-4.51(m，1H)，4.05-4.01(m，3H)，3.96-3.92(m，1H)，3.90(s，3H)，3.44-3.37(m，1H)，3.35(s，3H)，3.11-3.06(m，1H)，2.91(s，2H)，2.24-2.23(m，2H)，0.97(s，3H)，0.42(s，3H)。ESI-MS m/z444.2(M+H)+ Compound I-52-rac: Compound 52k-rac (75 mg, 0.1 mmol) was dissolved in 3 mL of tetrahydrofuran, and sodium hydroxide 32 mg and water 1 mL were added, and the reaction was stirred at 35 ° C for 2 hours. After the completion of the reaction, the pH was adjusted to about 2-3 with 1N hydrochloric acid and extracted with dichloromethane (20 mL×3). The organic phase was combined, dried over anhydrous sodium sulfate, Yield: 67.5%. 1 H-NMR (CDCl 3 , 400 MHz) δ: 8.58 (s, 1H), 7.15 (s, 1H), 7.06 (s, 1H), 6.66 (s, 1H), 5.05-5.03 (m, 1H), 4.52 4.51 (m, 1H), 4.05-4.01 (m, 3H), 3.96-3.92 (m, 1H), 3.90 (s, 3H), 3.44-3.37 (m, 1H), 3.35 (s, 3H), 3.11 3.06 (m, 1H), 2.91 (s, 2H), 2.24 - 2.23 (m, 2H), 0.97 (s, 3H), 0.42 (s, 3H). ESI-MS m/z 444.2 (M+H) +

实施例34.制备10-甲氧基-6-(1-甲氧基-2-甲基丙-2-基)-9-((1R，3R)-3-甲氧基环丁氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-53-rac)Example 34. Preparation of 10-methoxy-6-(1-methoxy-2-methylpropan-2-yl)-9-((1R,3R)-3-methoxycyclobutoxy) 2-oxo-6,7-dihydro-2H-pyrido[2,1-a]isoquinoline-3-carboxylic acid (I-53-rac)

Figure PCTCN2017094946-appb-000047
Figure PCTCN2017094946-appb-000047

化合物53a-rac：将化合物52j-rac(100mg，0.25mmol)，化合物5c(90mg，0.50mmol)和碳酸钾(103mg，0.75mmol)加入到5mL DMF中，置换氮气3次，于90℃加热搅拌反应5小时。反应完毕后加水稀释，用乙酸乙酯萃取(20mL×3)，合并有机相，无水硫酸钠干燥，浓缩得粗产品120mg。Compound 53a-rac: Compound 52j-rac (100 mg, 0.25 mmol), compound 5c (90 mg, 0.50 mmol) and potassium carbonate (103 mg, 0.75 mmol) were added to 5 mL of DMF, replaced with nitrogen three times and heated and stirred at 90 °C. Reaction for 5 hours. After completion of the reaction, the mixture was diluted with EtOAc (EtOAc m.

化合物I-53-rac：将化合物53a-rac(120mg，0.25mmol)溶于3mL四氢呋喃中，加入氢氧化钠32mg和水1mL，于35℃搅拌反应2小时。反应完毕后用1N盐酸调pH至2-3左右，二氯甲烷萃取(20mL×3)，合并有机相，无水硫酸钠干燥，浓缩后薄层层析得产品20mg。收率：17.5％。1H-NMR(CDCl3，400MHz)δ：8.58(s，1H)，7.15(s，1H)，7.06(s，1H)，6.52(s，1H)，5.00-4.90(m，1H)，4.52-4.50(m，1H)，4.20(m，1H)，3.97(m，1H)，3.93(s，3H)，3.64(m，1H)，3.36(s，3H)，3.31(s，3H)，3.08(m，1H)，2.91(m，2H)，2.59(m，4H)，0.88(s，3H)，0.41(s，3H)。ESI-MS m/z 458.2(M+H)+ Compound I-53-rac: Compound 53a-rac (120 mg, 0.25 mmol) was dissolved in 3 mL of tetrahydrofuran, and sodium hydroxide 32 mg and water 1 mL were added, and the reaction was stirred at 35 ° C for 2 hours. After the completion of the reaction, the pH was adjusted to about 2-3 with 1N hydrochloric acid, and extracted with dichloromethane (20 mL×3). The organic phase was combined and dried over anhydrous sodium sulfate. Yield: 17.5%. 1 H-NMR (CDCl 3 , 400 MHz) δ: 8.58 (s, 1H), 7.15 (s, 1H), 7.06 (s, 1H), 6.52 (s, 1H), 5.00 - 4.90 (m, 1H), 4.52 4.50 (m, 1H), 4.20 (m, 1H), 3.97 (m, 1H), 3.93 (s, 3H), 3.64 (m, 1H), 3.36 (s, 3H), 3.31 (s, 3H), 3.08 (m, 1H), 2.91 (m, 2H), 2.59 (m, 4H), 0.88 (s, 3H), 0.41 (s, 3H). ESI-MS m/z 458.2(M+H) +

实施例35.制备10-甲氧基-6-(1-甲氧基-2-甲基丙-2-基)-9-((1-(甲氧基甲基)环丙基)甲氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-54-rac)Example 35. Preparation of 10-methoxy-6-(1-methoxy-2-methylpropan-2-yl)-9-((1-(methoxymethyl)cyclopropyl)methoxy 2-oxo-6,7-dihydro-2H-pyrido[2,1-a]isoquinoline-3-carboxylic acid (I-54-rac)

Figure PCTCN2017094946-appb-000048
Figure PCTCN2017094946-appb-000048

化合物54a-rac：将化合物52j-rac(100mg，0.25mmol)，化合物6c(97mg，0.50mmol)和碳酸钾(103mg，0.75mmol) 加入到5mL DMF中，置换氮气3次，于90℃加热搅拌反应5小时。反应完毕后加水稀释，用乙酸乙酯萃取(20mL×3)，合并有机相，无水硫酸钠干燥，浓缩得粗产品110mg。Compound 54a-rac: Compound 52j-rac (100 mg, 0.25 mmol), Compound 6c (97 mg, 0.50 mmol) and potassium carbonate (103 mg, 0.75 mmol) After adding to 5 mL of DMF, the nitrogen gas was replaced three times, and the reaction was stirred under heating at 90 ° C for 5 hours. After completion of the reaction, the mixture was diluted with EtOAc (EtOAc)EtOAc.

化合物I-54-rac：将化合物54a-rac(110mg，0.22mmol)溶于3mL四氢呋喃中，加入氢氧化钠40mg和水1mL，于35℃搅拌反应2小时。反应完毕后用1N盐酸调pH至2-3左右，二氯甲烷萃取(20mL×3)，合并有机相，无水硫酸钠干燥，浓缩后薄层层析得产品27mg。收率：26.1％。1H-NMR(CDCl3，400MHz)δ：8.57(s，1H)，7.13(s，1H)，7.05(s，1H)，6.74(s，1H)，4.49(d，J＝6.8Hz，1H)，3.99(s，2H)，3.95(d，J＝9.2Hz，1H)，3.90(s，3H)，3.41(s，2H)，3.36(d，J＝4.8Hz，6H)，3.07(d，J＝17.6Hz，1H)，2.91(s，2H)，0.97(s，3H)，0.71-0.63(m，4H)，0.43(s，3H)；ESI-MS m/z 472.2(M+H)+ Compound I-54-rac: Compound 54a-rac (110 mg, 0.22 mmol) was dissolved in 3 mL of tetrahydrofuran, 40 mg of sodium hydroxide and 1 mL of water were added, and the reaction was stirred at 35 ° C for 2 hours. After completion of the reaction, the pH was adjusted to about 2-3 with 1N hydrochloric acid, and extracted with dichloromethane (20 mL×3). The organic phase was combined and dried over anhydrous sodium sulfate. Yield: 26.1%. 1 H-NMR (CDCl 3 , 400 MHz) δ: 8.57 (s, 1H), 7.13 (s, 1H), 7.05 (s, 1H), 6.74 (s, 1H), 4.49 (d, J = 6.8 Hz, 1H) ), 3.99 (s, 2H), 3.95 (d, J = 9.2 Hz, 1H), 3.90 (s, 3H), 3.41 (s, 2H), 3.36 (d, J = 4.8 Hz, 6H), 3.07 (d) , J = 17.6 Hz, 1H), 2.91 (s, 2H), 0.97 (s, 3H), 0.71 - 0.63 (m, 4H), 0.43 (s, 3H); ESI-MS m/z 472.2 (M+H ) +

实施例36.制备10-甲氧基-6-(1-甲氧基-2-甲基丙-2-基)-9-((3-(甲氧基甲基)氧杂环丁烷-3-基)甲氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-55-rac)Example 36. Preparation of 10-methoxy-6-(1-methoxy-2-methylpropan-2-yl)-9-((3-(methoxymethyl)oxetane- 3-yl)methoxy)-2-oxo-6,7-dihydro-2H-pyrido[2,1-a]isoquinoline-3-carboxylic acid (I-55-rac)

Figure PCTCN2017094946-appb-000049
Figure PCTCN2017094946-appb-000049

化合物55a-rac：将化合物67j-rac(54mg，0.13mmol)，化合物7e(56mg，0.26mmol)和碳酸钾(56mg，0.39mmol)加入到5mL DMF中，置换氮气3次，于90℃加热搅拌反应5小时。反应完毕后加水稀释，用乙酸乙酯萃取(20mL×3)，合并有机相，无水硫酸钠干燥，浓缩得粗品100mg。Compound 55a-rac: Compound 67j-rac (54 mg, 0.13 mmol), compound 7e (56 mg, 0.26 mmol) and potassium carbonate (56 mg, 0.39 mmol) were added to 5 mL of DMF, replaced with nitrogen three times and heated and stirred at 90 °C. Reaction for 5 hours. After completion of the reaction, the mixture was diluted with EtOAc (EtOAc m.

化合物I-55-rac：将化合物55a-rac(100mg，0.19mmol)溶于4mL四氢呋喃中，加入氢氧化钠46mg和水1mL，于35℃搅拌反应2小时。反应完毕后用1N盐酸调pH至2-3左右，二氯甲烷萃取(20mL×3)，合并有机相，无水硫酸钠干燥，浓缩后薄层层析得产品10mg。收率：10.8％。1H-NMR(CDCl3，400MHz)δ：8.58(s，1H)，7.15(s，1H)，7.05(s，1H)，6.79(s，1H)，4.64-4.51(m，5H)，4.30-4.24(m，2H)，3.89(s，3H)，3.81-3.74(m，2H)，3.39(d，7H，J＝18.8Hz)，3.12-3.08(m，1H)，2.92(s，2H)，0.98(s，3H)，0.43(s，3H)。ESI-MS m/z 488.2(M+H)+ Compound I-55-rac: Compound 55a-rac (100 mg, 0.19 mmol) was dissolved in 4 mL of tetrahydrofuran, and sodium hydroxide (46 mg) and water (1 mL) were added, and the mixture was stirred at 35 ° C for 2 hours. After the completion of the reaction, the pH was adjusted to about 2-3 with 1N hydrochloric acid, and extracted with dichloromethane (20 mL×3). The organic phase was combined and dried over anhydrous sodium sulfate. Yield: 10.8%. 1 H-NMR (CDCl 3 , 400 MHz) δ: 8.58 (s, 1H), 7.15 (s, 1H), 7.05 (s, 1H), 6.79 (s, 1H), 4.64 - 4.51 (m, 5H), 4.30 -4.24 (m, 2H), 3.89 (s, 3H), 3.81-3.74 (m, 2H), 3.39 (d, 7H, J = 18.8 Hz), 3.12-3.08 (m, 1H), 2.92 (s, 2H) ), 0.98 (s, 3H), 0.43 (s, 3H). ESI-MS m/z 488.2(M+H) +

实施例37.制备10-甲氧基-6-(1-甲氧基-2-甲基丙-2-基)-9-((3-甲氧基环丁基)甲氧基)-2-氧代-6，7-二氢-2H-吡啶并[2，1-a]异喹啉-3-羧酸(I-56-rac)Example 37. Preparation of 10-methoxy-6-(1-methoxy-2-methylpropan-2-yl)-9-((3-methoxycyclobutyl)methoxy)-2 -oxo-6,7-dihydro-2H-pyrido[2,1-a]isoquinoline-3-carboxylic acid (I-56-rac)

Figure PCTCN2017094946-appb-000050
Figure PCTCN2017094946-appb-000050

化合物56a-rac：将化合物52j-rac(100mg，0.25mmol)，化合物8d(96mg，0.50mmol)和碳酸钾(103mg，0.75mmol)加入到5mL DMF中，置换氮气3次，于90℃加热搅拌反应5小时。反应完毕后加水稀释，用乙酸乙酯萃取(20mL×3)，合并有机相，无水硫酸钠干燥，浓缩后薄层层析得产品60mg。收率：40.0％。Compound 56a-rac: Compound 52j-rac (100 mg, 0.25 mmol), compound 8d (96 mg, 0.50 mmol) and potassium carbonate (103 mg, 0.75 mmol) were added to 5 mL of DMF, replaced with nitrogen three times and heated and stirred at 90 °C. Reaction for 5 hours. After completion of the reaction, it was diluted with water and extracted with ethyl acetate (20 mL×3). The organic phase was combined and dried over anhydrous sodium sulfate. Yield: 40.0%.

化合物I-56-rac：将化合物56a-rac(60mg，0.1mmol)溶于4mL四氢呋喃中，加入氢氧化钠25mg和水1mL，于35℃搅拌反应2小时。反应完毕后用1N盐酸调pH至2-3左右，二氯甲烷萃取(20mL×3)，合并有机相，无水硫酸钠干燥，浓缩后薄层层析得产品40mg。收率：84.7％。1H-NMR(CDCl3，400MHz)δ：8.57(s，1H)，7.13(s，1H)，7.04(s，1H)，6.70(s，1H)，4.51-4.50(m，1H)，4.07-4.05(m，2H)，3.90(s，3H)，3.84(t，J＝7.2Hz，1H)，3.36(s，3H)，3.25(s，3H)，3.06(m，1H)，2.91(s，2H)，2.53-2.48(m，1H)，2.42-2.38(m，1H)，2.25-2.21(m，1H)，1.82-1.77(m，2H)，0.98(s，3H)，0.42(s，3H)；ESI-MS m/z 472.2(M+H)+ Compound I-56-rac: Compound 56a-rac (60 mg, 0.1 mmol) was dissolved in 4 mL of tetrahydrofuran, 25 mg of sodium hydroxide and 1 mL of water were added, and the reaction was stirred at 35 ° C for 2 hours. After the completion of the reaction, the pH was adjusted to about 2-3 with 1N hydrochloric acid, and extracted with dichloromethane (20 mL×3). The organic phase was combined, dried over anhydrous sodium sulfate, Yield: 84.7%. 1 H-NMR (CDCl 3 , 400 MHz) δ: 8.57 (s, 1H), 7.13 (s, 1H), 7.04 (s, 1H), 6.70 (s, 1H), 4.51-4.50 (m, 1H), 4.07- 4.05 (m, 2H), 3.90 (s, 3H), 3.84 (t, J = 7.2 Hz, 1H), 3.36 (s, 3H), 3.25 (s, 3H), 3.06 (m, 1H), 2.91 (s) , 2H), 2.53-2.48 (m, 1H), 2.42 - 2.38 (m, 1H), 2.25 - 2.21 (m, 1H), 1.82-1.77 (m, 2H), 0.98 (s, 3H), 0.42 (s) , 3H); ESI-MS m/z 472.2 (M+H) +

实施例38.体外生物活性研究 Example 38. In vitro biological activity study

待测化合物：Test compound:

本发明的化合物：化合物I-2、化合物I-5、化合物I-6、化合物I-7、化合物I-8、化合物I-9、化合物I-10、化合物I-11、化合物I-12、化合物I-19、化合物I-20、化合物I-1-rac、化合物I-2-rac、化合物I-3-rac、化合物I-4-rac、化合物I-5-rac、化合物I-6-rac、化合物I-7-rac、化合物I-8-rac、化合物I-9-rac、化合物I-10-rac、化合物I-11-rac、化合物I-22-rac、化合物I-23-rac、化合物I-24-rac、化合物I-25-rac、化合物I-26-rac、化合物I-28-rac、化合物I-29-rac、化合物I-30-rac、化合物I-31-rac、化合物I-32-rac、化合物I-52-rac、化合物I-53-rac、化合物I-54-rac、化合物I-55-rac、化合物I-56-rac；Compounds of the invention: Compound 1-2, Compound 1-5, Compound 1-6, Compound -7, Compound VIII, Compound -9, Compound 1-10, Compound 1-1, Compound 1-1, Compound I-19, Compound I-20, Compound I-1-rac, Compound I-2-rac, Compound I-3-rac, Compound I-4-rac, Compound I-5-rac, Compound I-6- Rac, compound I-7-rac, compound I-8-rac, compound I-9-rac, compound I-10-rac, compound I-11-rac, compound I-22-rac, compound I-23-rac , compound I-24-rac, compound I-25-rac, compound I-26-rac, compound I-28-rac, compound I-29-rac, compound I-30-rac, compound I-31-rac, Compound I-32-rac, compound I-52-rac, compound I-53-rac, compound I-54-rac, compound I-55-rac, compound I-56-rac;

对照化合物：A、A-rac(结构式如下所示)：Control compound: A, A-rac (structure is shown below):

Figure PCTCN2017094946-appb-000051
Figure PCTCN2017094946-appb-000051

试验方法：在96孔板内，将HepG2.2.15细胞系按照1.5×104个细胞/孔铺板。第二天加入化合物处理细胞，待测化合物3倍稀释，8个浓度点，平行测定2复孔。培养液中DMSO的终浓度为0.5％。第五天更换新的含化合物的培养液，第八天收集上清，用ELISA法检测细胞上清中HBsAg。相对空白对照计算抑制百分率。结果参见表1。Test method: HepG2.2.15 cell line was plated at 1.5 x 104 cells/well in a 96-well plate. The compound was treated with the compound on the next day, and the test compound was diluted 3 times, 8 concentration points, and 2 duplicate wells were determined in parallel. The final concentration of DMSO in the culture broth was 0.5%. On the fifth day, a new culture solution containing the compound was replaced, and on the eighth day, the supernatant was collected, and HBsAg in the cell supernatant was detected by ELISA. Percent inhibition was calculated relative to the blank control. See Table 1 for the results.

表1.化合物对于HBsAg的抑制活性Table 1. Inhibitory activity of compounds against HBsAg

化合物Compound EC50(nM)EC 50 (nM) 化合物Compound EC50(nM)EC 50 (nM) 化合物Compound EC50(nM)EC 50 (nM) 化合物Compound EC50(nM)EC 50 (nM) I-2I-2 1.821.82 I-1-racI-1-rac 5.745.74 I-22-racI-22-rac 5.465.46 I-53-racI-53-rac 6.656.65 I-5I-5 1.611.61 I-2-racI-2-rac 3.263.26 I-23-racI-23-rac 6.836.83 I-54-racI-54-rac 5.345.34 I-6I-6 3.153.15 I-3-racI-3-rac 7.047.04 I-24-racI-24-rac 5.785.78 I-55-racI-55-rac 7.237.23 I-7I-7 3.823.82 I-4-racI-4-rac 3.623.62 I-25-racI-25-rac 6.976.97 I-56-racI-56-rac 5.955.95 I-8I-8 1.541.54 I-5-racI-5-rac 3.563.56 I-26-racI-26-rac 7.567.56 AA 4.064.06 I-9I-9 3.263.26 I-6-racI-6-rac 4.124.12 I-28-racI-28-rac 5.785.78 A-racA-rac 8.218.21 I-10I-10 3.013.01 I-7-racI-7-rac 4.684.68 I-29-racI-29-rac 6.456.45     I-11I-11 2.572.57 I-8-racI-8-rac 5.595.59 I-30-racI-30-rac 6.746.74     I-12I-12 3.833.83 I-9-racI-9-rac 3.713.71 I-31-racI-31-rac 5.985.98     I-19I-19 2.892.89 I-10-racI-10-rac 3.783.78 I-32-racI-32-rac 7.317.31     I-20I-20 2.562.56 I-11-racI-11-rac 2.612.61 I-52-racI-52-rac 7.577.57    

结论：大多数合成的化合物具有优秀的抑制HBsAg的活性，活性低于10nM。Conclusion: Most of the synthesized compounds have excellent activity in inhibiting HBsAg with an activity lower than 10 nM.

实施例39.大鼠PK研究Example 39. Rat PK study

静脉注射给药制剂配制：准确称量样品2～3mg，加入适量N，N-二甲基乙酰胺(DMA)，涡旋振荡使固体物质完全溶解；再加入适量体积的30％solutol HS-15水溶液，涡旋振荡后再加入saline，使得DMA∶30％solutol HS-15∶saline＝20∶20∶60(v/v/v)，涡旋振荡使液体混合均匀，并过滤，得浓度为0.4mg.mL-1的给药制剂。Formulation for intravenous administration: accurately weigh 2 to 3 mg of the sample, add appropriate amount of N,N-dimethylacetamide (DMA), vortex to completely dissolve the solid matter; then add an appropriate volume of 30% solutol HS-15 The aqueous solution was added to the saline after vortexing, so that DMA: 30% solutol HS-15:saline=20:20:60 (v/v/v), the liquid was uniformly mixed by vortexing, and filtered to obtain a concentration of 0.4. A preparation for administration of mg.mL -1 .

口服给药制剂配制：准确称量样品10mg，加入适量体积的0.5％CMC-Na水溶液，涡旋振荡后超声至混合均匀，得浓度为1mg.mL-1的给药制剂。Formulation for oral administration: 10 mg of the sample was accurately weighed, an appropriate volume of 0.5% CMC-Na aqueous solution was added, and the mixture was vortexed and ultrasonicated until homogeneously mixed to obtain a preparation having a concentration of 1 mg.mL -1 .

给药制剂均于给药当天新鲜配制，并进行留样，对留样进行给药制剂实际浓度的测定。The preparations for administration were freshly prepared on the day of administration, and samples were taken for the determination of the actual concentration of the preparation for administration.

A组S-D大鼠分别单次静脉注射(IV)给予2mg·kg-1的给药制剂；B组S-D大鼠单次灌胃(PO)给予10mg·kg-1的给药制剂。分别于给药前及给药后5分钟(仅静脉注射组)、15分钟、30分钟、1小时、2小时、4小时、8小时、12小时和24小时，由颈静脉采血0.15mL，置于EDTA-K2抗凝血管中。所有全血样品离心(5500转/分)10分钟后分离血浆，保存在-30～-10℃的冰箱中。利用LC-MS/MS分析方法，测定S-D大鼠血浆中待测化合物的浓度。采用Pharsight Phoenix 7.0中的非房室模型计算相应的药代动力学参数。结果参见表2a和2b。 Group A SD rats were given a single intravenous injection (IV) with a dose of 2 mg·kg -1 , and group B SD rats were given a single dose (PO) with a dose of 10 mg·kg -1 . 0.15 mL of blood was collected from the jugular vein before administration and 5 minutes after administration (intravenous group only), 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours and 24 hours. In the EDTA-K2 anticoagulation tube. All whole blood samples were centrifuged (5500 rpm) for 10 minutes, and the plasma was separated and stored in a refrigerator at -30 to -10 °C. The concentration of the test compound in the plasma of SD rats was determined by LC-MS/MS analysis. The corresponding pharmacokinetic parameters were calculated using a non-compartmental model in Pharsight Phoenix 7.0. See Tables 2a and 2b for the results.

表2a.测试化合物的PK参数(静脉注射)Table 2a. PK parameters of test compounds (intravenous)

Figure PCTCN2017094946-appb-000052
Figure PCTCN2017094946-appb-000052

表2b 测试化合物的PK参数(灌胃)Table 2b PK parameters of test compounds (gavage)

Figure PCTCN2017094946-appb-000053
Figure PCTCN2017094946-appb-000053

结论：综合静脉注射和灌胃给药的PK数据，本发明化合物(尤其是I-2，I-5，I-6，I-8)具有良好的PK特性，显示很好的开发前景。可能是由于在化学结构上侧链的环化改善了化合物的内在代谢稳定性。Conclusion: The compounds of the present invention (especially I-2, I-5, I-6, I-8) have good PK characteristics, and show good development prospects, combined with PK data for intravenous and intragastric administration. This may be due to the fact that the cyclization of the side chains on the chemical structure improves the intrinsic metabolic stability of the compound.

以上实施例的说明只是用于帮助理解本发明的方法及其核心思想。应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以对本发明进行若干改进和修饰，这些改进和修饰也落入本发明权利要求的保护范围内。 The above description of the embodiments is merely to assist in understanding the method of the present invention and its core idea. It should be noted that those skilled in the art can make various modifications and changes to the present invention without departing from the spirit and scope of the invention.

Claims (19)

一种式(I)所示的异喹啉酮类化合物或其立体异构体、可药用盐、溶剂化物或结晶，An isoquinolinone compound of the formula (I) or a stereoisomer thereof, a pharmaceutically acceptable salt, a solvate or a crystal thereof,
Figure PCTCN2017094946-appb-100001
Figure PCTCN2017094946-appb-100001
 其中：among them: (1)R1选自H、氘、C1-6烷基、氰基、卤素、羧基、酯基、C3-6环烷基、C4-8杂环烷基、卤代C1-6烷基或C6-10芳基；(1) R 1 is selected from the group consisting of H, hydrazine, C 1-6 alkyl, cyano, halogen, carboxyl, ester, C 3-6 cycloalkyl, C 4-8 heterocycloalkyl, halogen C 1- 6 alkyl or C 6-10 aryl; (2)R2选自卤素、C1-3烷氧基、氘代C1-3烷氧基、C1-6烷基、C3-6环烷基、C3-6环烷基氧基、C4-8环杂烷基C1-6烷基、卤代C1-3烷基氧基、卤代C3-6环烷基、C3-6环烷基C1-6烷基或R2和R3以碳原子连接形成环；(2) R 2 is selected from the group consisting of halogen, C 1-3 alkoxy, deuterated C 1-3 alkoxy, C 1-6 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyloxy , C 4-8 cycloheteroalkyl C 1-6 alkyl, halo C 1-3 alkyloxy, halo C 3-6 cycloalkyl, C 3-6 cycloalkyl C 1-6 alkane a group or R 2 and R 3 are bonded by a carbon atom to form a ring; (3)R3为(a)具有环结构和/或不饱和键的C4-12烃基，该C4-12烃基的氢原子未被取代，或为选自氘、卤素、氰基、羟基、巯基中的一个或多个取代基取代，且该C4-12烃基未被杂原子间断，或被O，S，NH，C＝O，C＝S，O＝S＝O中的一个或多个所间断，所述杂原子选自氧、硫或氮；或者，(b)R2和R3以碳原子连接形成环；(3) R 3 is (a) a C 4-12 hydrocarbon group having a ring structure and/or an unsaturated bond, the hydrogen atom of the C 4-12 hydrocarbon group is unsubstituted, or is selected from the group consisting of ruthenium, halogen, cyano, and hydroxy Substituting one or more substituents in the fluorenyl group, and the C 4-12 hydrocarbon group is not interrupted by a hetero atom, or is one of O, S, NH, C=O, C=S, O=S=O or a plurality of discontinuities, the hetero atom is selected from oxygen, sulfur or nitrogen; or, (b) R 2 and R 3 are bonded by a carbon atom to form a ring; (4)R4选自氢、氘、卤素、氰基、酯基或C1-3烷基；(4) R 4 is selected from the group consisting of hydrogen, hydrazine, halogen, cyano, ester or C 1-3 alkyl; (5)R5，R5′独立的选自氢、氘、卤素、甲基、甲氧基或R5，R5′形成碳环或杂环；或R5，R6形成碳环或杂环；(5) R 5 , R 5 ' independently selected from hydrogen, hydrazine, halogen, methyl, methoxy or R 5 , R 5 'forms a carbocyclic or heterocyclic ring; or R 5 , R 6 form a carbocyclic or heterocyclic ring ring; (6)M为CH或N；(6) M is CH or N; (7)R6选自C1-6烷基、C1-6烷氧C1-6烷基、羟基C1-6烷基、芳基、卤代C1-6烷基或C3-6环烷基C1-6烷基；(7) R 6 is selected from C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, hydroxy C 1-6 alkyl, aryl, halogen C 1-6 alkyl or C 3- 6 cycloalkyl C 1-6 alkyl; (8)W为N或CR7，其中R7选自氢、氘、羟基、卤素、C1-3烷基、C1-6烷氧基、C3-6环烷基氧基、酯基、羧基或氰基；(8) W is N or CR 7 , wherein R 7 is selected from the group consisting of hydrogen, hydrazine, hydroxy, halogen, C 1-3 alkyl, C 1-6 alkoxy, C 3-6 cycloalkyloxy, ester , carboxyl or cyano; (9)R8选自羧基、酯基、C1-6烷基、C3-6环烷基、C1-6烷基炔基或C3-6环烷基炔基，其中所述酯基的烷基部分选自C1-6烷基、C3-8环烷基、C3-8环烷基炔基、C1-6烷基炔基、苄基、C1-6烷基C(O)O-C1-3烷基、C1-6烷基-OC(O)O-C1-3烷基。(9) R 8 is selected from a carboxyl group, an ester group, a C 1-6 alkyl group, a C 3-6 cycloalkyl group, a C 1-6 alkyl alkynyl group or a C 3-6 cycloalkyl alkynyl group, wherein the ester The alkyl moiety of the group is selected from the group consisting of C 1-6 alkyl, C 3-8 cycloalkyl, C 3-8 cycloalkyl alkynyl, C 1-6 alkyl alkynyl, benzyl, C 1-6 alkyl C(O)OC 1-3 alkyl, C 1-6 alkyl-OC(O)OC 1-3 alkyl. 根据权利要求1所述的异喹啉酮类化合物或其立体异构体、可药用盐、溶剂化物或结晶，其特征在于，所述(a)中，所述环结构为3～8元环；所述不饱和键为双键或三键。The isoquinolinone compound or a stereoisomer, pharmaceutically acceptable salt, solvate or crystal thereof according to claim 1, wherein in the (a), the ring structure is 3 to 8 yuan. a ring; the unsaturated bond is a double bond or a triple bond. 根据权利要求1所述的异喹啉酮类化合物或其立体异构体、可药用盐、溶剂化物或结晶，其特征在于，所述(a)中，所述的环结构为饱和环。The isoquinolinone compound or a stereoisomer thereof, a pharmaceutically acceptable salt, a solvate or a crystal thereof according to claim 1, wherein in the above (a), the ring structure is a saturated ring. 根据权利要求1所述的异喹啉酮类化合物或其立体异构体、可药用盐、溶剂化物或结晶，其特征在于，所述(a)中，所述环结构、不饱和键的个数分别为1～2个。The isoquinolinone compound or a stereoisomer, pharmaceutically acceptable salt, solvate or crystal thereof according to claim 1, wherein in the above (a), the ring structure, an unsaturated bond The number is 1 to 2 respectively. 根据权利要求1所述的异喹啉酮类化合物或其立体异构体、可药用盐、溶剂化物或结晶，其特征在于，所述(a)具有一个3～8元饱和碳环或3～8元饱和杂环、至少一个杂原子或至少一个双键或三键。The isoquinolinone compound or a stereoisomer, pharmaceutically acceptable salt, solvate or crystal thereof according to claim 1, wherein the (a) has a 3 to 8 membered saturated carbocyclic ring or 3 ～8-membered saturated heterocyclic ring, at least one hetero atom or at least one double or triple bond. 根据权利要求1至5中任一项权利要求所述的异喹啉酮类化合物或其立体异构体、可药用盐、溶剂化物或结晶，其特征在于，所述(a)中，所述环结构、不饱和键以及杂原子中的至少二者同时存在。The isoquinolinone compound or a stereoisomer, pharmaceutically acceptable salt, solvate or crystal thereof according to any one of claims 1 to 5, wherein in (a), At least two of the ring structure, the unsaturated bond, and the hetero atom are present at the same time. 根据权利要求6所述的异喹啉酮类化合物或其立体异构体、可药用盐、溶剂化物或结晶，其特征在于，所述(a)为满足如下任意一项所述条件的基团：The isoquinolinone compound or a stereoisomer, pharmaceutically acceptable salt, solvate or crystal thereof according to claim 6, wherein the (a) is a group satisfying the conditions described in any one of the following group: a1)同时具有所述的环结构和碳碳不饱和键，且环结构和碳碳不饱和键分别有且只有一个；A1) having both the ring structure and the carbon-carbon unsaturated bond, and having one and only one ring structure and carbon-carbon unsaturated bond; a2)同时具有所述的环结构和1～3个杂原子，且所述杂原子中至少一个为氧，且该氧原子通过单键与式(I)中 苯环连接；A2) having both the ring structure and 1 to 3 hetero atoms, and at least one of the hetero atoms is oxygen, and the oxygen atom is passed through a single bond with the formula (I) Benzene ring connection; a3)同时具有不饱和键和1～3个杂原子，其中不饱和键为碳碳双键，碳碳三键或者碳氧双键，当不饱和键为碳碳双键或碳碳三键时，它们的一端优选通过单键与式(I)中苯环连接。A3) has both an unsaturated bond and 1 to 3 heteroatoms, wherein the unsaturated bond is a carbon-carbon double bond, a carbon-carbon triple bond or a carbon-oxygen double bond, when the unsaturated bond is a carbon-carbon double bond or a carbon-carbon triple bond One end of them is preferably linked to the benzene ring of the formula (I) by a single bond. 根据权利要求1所述的异喹啉酮类化合物或其立体异构体、可药用盐、溶剂化物或结晶，其特征在于，R3为C5-11双环烷基；C3-6环烷基炔基；C3-6环烷基烯基；C1-3烷氧C1-6烷基炔基；C1-3烷氧C1-6烷基烯基；C4-8环杂烷基；或者，The isoquinolinone compound or a stereoisomer, pharmaceutically acceptable salt, solvate or crystal thereof according to claim 1, wherein R 3 is a C 5-11 bicycloalkyl group; C 3-6 ring Alkynyl group; C 3-6 cycloalkylalkenyl; C 1-3 alkoxy C 1-6 alkyl alkynyl; C 1-3 alkoxy C 1-6 alkylalkenyl; C 4-8 ring Heteroalkyl; or, R3为RA-O-，其中RA选自C3-8环烷基；C5-11双环烷基；氘代C1-6烷基；C4-8环杂烷基；C1-6烷基羰基C1-6烷基；氘代C1-3烷氧基C1-6烷基；C1-3烷氧基C3-8环烷基；C1-3烷氧基C3-8环烷基C1-6烷基；C3-8杂环烷基；C1-3烷氧基C1-6烷基，其中烷基被C3-8环烷烃或C4-8杂环烷烃取代，杂环烷烃的杂原子选自氧、硫或氮；当RA选自C1-3烷氧基C1-6烷基时，R5、R5′独立选自氘、氟、氯、羟基、氰基，且W为N或CR7，其中R7选自氘、氟、氯、羟基、氰基。R 3 is R A -O-, wherein R A is selected from C 3-8 cycloalkyl; C 5-11 bicycloalkyl; deuterated C 1-6 alkyl; C 4-8 cycloheteroalkyl; C 1 -6 alkylcarbonyl C 1-6 alkyl; deuterated C 1-3 alkoxy C 1-6 alkyl; C 1-3 alkoxy C 3-8 cycloalkyl; C 1-3 alkoxy C 3-8 cycloalkyl C 1-6 alkyl; C 3-8 heterocycloalkyl; C 1-3 alkoxy C 1-6 alkyl, wherein alkyl is C 3-8 cycloalkane or C 4 a heterocyclic alkane substituted, the hetero atom of the heterocycloalkane is selected from oxygen, sulfur or nitrogen; when R A is selected from C 1-3 alkoxy C 1-6 alkyl, R 5 , R 5 ' are independently selected from Anthracene, fluorine, chlorine, hydroxyl, cyano, and W is N or CR 7 , wherein R 7 is selected from the group consisting of hydrazine, fluorine, chlorine, hydroxyl, and cyano. 根据权利要求1所述的异喹啉酮类化合物或其立体异构体、可药用盐、溶剂化物或结晶，其特征在于，所述R3选自C3-8环烷氧基、C3-8杂环烷氧基、C1-3烷氧C3-8环烷氧基、C1-3烷氧C3-8环烷基C1-6烷氧基、C3-8杂环烷基、C1-3烷氧C2-9烯基、C1-3烷氧C2-9炔基、C3-8环烷基C2-9烯基、C3-8环烷基C2-9炔基。The isoquinolinone compound or a stereoisomer, pharmaceutically acceptable salt, solvate or crystal thereof according to claim 1, wherein the R 3 is selected from the group consisting of C 3-8 cycloalkoxy, C 3-8 heterocycloalkoxy, C 1-3 alkoxy C 3-8 cycloalkoxy, C 1-3 alkoxy C 3-8 cycloalkyl C 1-6 alkoxy, C 3-8 Cycloalkyl, C 1-3 alkoxy C 2-9 alkenyl, C 1-3 alkoxy C 2-9 alkynyl, C 3-8 cycloalkyl C 2-9 alkenyl, C 3-8 naphthenic Alkyl C 2-9 alkynyl. 根据权利要求1所述的异喹啉酮类化合物或其立体异构体、可药用盐、溶剂化物或结晶，其特征在于，所述R2选自C1-3烷氧基、卤素、C3-6环烷基、苄基。The isoquinolinone compound or a stereoisomer, pharmaceutically acceptable salt, solvate or crystal thereof according to claim 1, wherein the R 2 is selected from the group consisting of C 1-3 alkoxy, halogen, C 3-6 cycloalkyl, benzyl. 根据权利要求1所述的异喹啉酮类化合物或其立体异构体、可药用盐、溶剂化物或结晶，其特征在于，R6选自甲基、乙基、异丙基、丁基、异丁基、甲氧基甲基、甲氧基乙基、甲氧基异丙基、甲氧基丁基、甲氧基异丁基、乙氧基甲基、乙氧基乙基、乙氧基异丙基、乙氧基丁基、乙氧基异丁基、羟基甲基、羟基乙基、羟基异丙基、羟基丁基、羟基异丁基。The isoquinolinone compound or a stereoisomer, pharmaceutically acceptable salt, solvate or crystal thereof according to claim 1, wherein R 6 is selected from the group consisting of methyl, ethyl, isopropyl and butyl. , isobutyl, methoxymethyl, methoxyethyl, methoxy isopropyl, methoxybutyl, methoxyisobutyl, ethoxymethyl, ethoxyethyl, B Oxypropyl, ethoxybutyl, ethoxyisobutyl, hydroxymethyl, hydroxyethyl, hydroxyisopropyl, hydroxybutyl, hydroxyisobutyl. 根据权利要求1所述的异喹啉酮类化合物或其立体异构体、可药用盐、溶剂化物或结晶，其特征在于，除去活泼氢之外，其余所有的氢原子都可以分别独立地被氘取代。The isoquinolinone compound or a stereoisomer thereof, a pharmaceutically acceptable salt, solvate or crystal thereof according to claim 1, wherein all of the hydrogen atoms except the active hydrogen are independently and independently Replaced by 氘. 根据权利要求1所述的异喹啉酮类化合物或其立体异构体、可药用盐、溶剂化物或结晶，其特征在于，所述的异喹啉酮类化合物选自如下化合物：The isoquinolinone compound or a stereoisomer, pharmaceutically acceptable salt, solvate or crystal thereof according to claim 1, wherein the isoquinolinone compound is selected from the group consisting of the following compounds:
Figure PCTCN2017094946-appb-100002
Figure PCTCN2017094946-appb-100002
Figure PCTCN2017094946-appb-100003
Figure PCTCN2017094946-appb-100003
Figure PCTCN2017094946-appb-100004
Figure PCTCN2017094946-appb-100004
 一种药物组合物，其特征在于：含有如权利要求1至13中任一项权利要求所述的式(I)所示的异喹啉酮类化合物、其立体异构体、可药用盐、溶剂化物或结晶，以及药学上可接受的载体或赋形剂，药物组合物的剂型优选为片剂、胶囊或针剂。A pharmaceutical composition comprising the isoquinolinone compound of the formula (I) according to any one of claims 1 to 13, a stereoisomer thereof, and a pharmaceutically acceptable salt thereof The solvate or crystallization, and the pharmaceutically acceptable carrier or excipient, the dosage form of the pharmaceutical composition is preferably a tablet, capsule or injection. 根据权利要求14所述的药物组合物，其特征在于：所述药物组合物为抗病毒药物组合物，其中还包含一种或多种治疗剂，所述治疗剂治选自以下组成的群：核苷类药物、利巴韦林、干扰素、HBV衣壳抑制剂(capsid inhibitor)、cccDNA形成抑制剂、cccDNA表观遗传修饰剂或乙肝RNAi药物、TLR7激动剂。The pharmaceutical composition according to claim 14, wherein the pharmaceutical composition is an antiviral pharmaceutical composition further comprising one or more therapeutic agents, the therapeutic agent being selected from the group consisting of: Nucleoside drugs, ribavirin, interferon, HBV capsid inhibitor, cccDNA formation inhibitor, cccDNA epigenetic modifier or hepatitis B RNAi drug, TLR7 agonist. 如权利要求1至13中任一项权利要求所述的式(I)所示的异喹啉酮类化合物、其立体异构体、可药用盐、溶剂化物或结晶或其与一种或多种选自核苷类药物、利巴韦林、干扰素、HBV衣壳抑制剂(capsid inhibitor)、cccDNA形成抑制剂、cccDNA表观遗传修饰剂或乙肝RNAi药物、TLR7激动剂的治疗剂的组合在制备预防和/或治疗病毒感染疾病药物，和/或乙肝表面抗原抑制剂(HBV Surface antigeninhibitors)和乙肝DNA抑制剂(HBV DNA production inhibitors)药物中的应用，所述病毒感染包括HBV或HDV的感染。The isoquinolinone compound of the formula (I), a stereoisomer, a pharmaceutically acceptable salt, a solvate or a crystal thereof, or a compound thereof, according to any one of claims 1 to 13 a plurality of therapeutic agents selected from the group consisting of nucleoside drugs, ribavirin, interferon, HBV capsid inhibitor, cccDNA formation inhibitor, cccDNA epigenetic modifier or hepatitis B RNAi drug, TLR7 agonist The use in combination for the preparation of a medicament for preventing and/or treating a viral infection, and/or a hepatitis B surface antigen inhibitor (HBV Surface antigen inhibitor) and a hepatitis B DNA production inhibitor (HBV DNA production inhibitors), the virus infection including HBV or HDV Infection. 一种制备权利要求1至13中任一项权利要求所述的式(I)所示的异喹啉酮类化合物、其立体异构体、可药用盐、溶剂化物或结晶的中间体，其特征在于：所述中间体如下式(II)所示：An isoquinolinone compound of the formula (I) according to any one of claims 1 to 13, a stereoisomer, a pharmaceutically acceptable salt, a solvate or a crystalline intermediate thereof, It is characterized in that the intermediate is represented by the following formula (II):
Figure PCTCN2017094946-appb-100005
式(II)中，所述R1、R2、R4、R5、R5’、R6、R8、W和N的定义同前述权利要求。
Figure PCTCN2017094946-appb-100005
In the formula (II), R 1 , R 2 , R 4 , R 5 , R 5 ', R 6 , R 8 , W and N are as defined in the preceding claims. 根据权利要求17所述的中间体，其特征在于：式(II)所示的中间体为化合物10或其异构体或消旋体The intermediate according to claim 17, wherein the intermediate represented by the formula (II) is the compound 10 or an isomer or racemate thereof.
Figure PCTCN2017094946-appb-100006
Figure PCTCN2017094946-appb-100006
 一种制备权利要求1至13中任一项权利要求所述的式(I)所示的异喹啉酮类化合物、其立体异构体、可药用盐、溶剂化物或结晶的制备方法，其特征在于：所述方法包括采取如权利要求17或18所述的式(II)所示的中间体，并使式(II)所示的中间体与RAOH、RAOMs或RABr反应，其中，当反应物为RAOH，所述反应采用Mitsunobu反应，在脱水剂为三苯基磷和/或偶氮二甲酸二异丙酯存在下进行；当反应物为RAOMs或RABr，所述反应为SN2反应，在碱为碳酸钾和/或碳酸铯以及催化量KI存在下进行。 A process for preparing an isoquinolinone compound represented by the formula (I) according to any one of claims 1 to 13, a stereoisomer, a pharmaceutically acceptable salt, a solvate or a crystal thereof, Characterized in that the method comprises the intermediate represented by formula (II) according to claim 17 or 18, and the intermediate represented by formula (II) and R A OH, R A OMs or R A Br reaction, wherein when the reactant is R A OH, the reaction is carried out using a Mitsunobu reaction in the presence of a dehydrating agent of triphenylphosphine and/or diisopropyl azodicarboxylate; when the reactant is R A OMs Or R A Br, the reaction is an SN 2 reaction, carried out in the presence of a base of potassium carbonate and/or cesium carbonate and a catalytic amount of KI.
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