WO2016206651A1 - 2-substituted benzimidazole-4-carboxamide compound and preparation method and use thereof - Google Patents

2-substituted benzimidazole-4-carboxamide compound and preparation method and use thereof Download PDF

Info

Publication number
WO2016206651A1
WO2016206651A1 PCT/CN2016/087368 CN2016087368W WO2016206651A1 WO 2016206651 A1 WO2016206651 A1 WO 2016206651A1 CN 2016087368 W CN2016087368 W CN 2016087368W WO 2016206651 A1 WO2016206651 A1 WO 2016206651A1
Authority
WO
WIPO (PCT)
Prior art keywords
substituted
unsubstituted
compound
mmol
nmr
Prior art date
Application number
PCT/CN2016/087368
Other languages
French (fr)
Chinese (zh)
Inventor
杨春皓
缪泽鸿
陈旭星
宦霞娟
王玉勤
谭村
陈奕
丁健
Original Assignee
中国科学院上海药物研究所
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 中国科学院上海药物研究所 filed Critical 中国科学院上海药物研究所
Priority to CN201680037353.6A priority Critical patent/CN107922430B/en
Publication of WO2016206651A1 publication Critical patent/WO2016206651A1/en

Links

Classifications

    • 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
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
    • C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D495/04—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

  • the invention belongs to the fields of medicinal chemistry and pharmacotherapy. Specifically relates to a class of 2-(thiophene or thiazolotetrahydropyridin-2-yl)-1H-benzimidazole-4-carboxamides or pharmaceutically acceptable salts thereof, processes for their preparation and their preparation and treatment
  • PARP poly ADP-ribose polymerase
  • the main DNA repair pathways include base-excision repair (BER), Nucleotide-excision repair (NER), mismatch repair (MMR), and homologous recombination (Homologous recombination, HR) and Nonhomologous end joining (NHEJ) and the like.
  • BER, NER and MMR are the main DNA single-strand break (SSB) repair pathways.
  • HR and NHEJ are the main DNA double-strand break (DSB) repair pathways.
  • HR is a reliable and accurate repair pathway, while NHEJ is unstable and easy. An error occurred.
  • some DNA damage can be directly repaired by the corresponding enzymes.
  • guanine methylation can be repaired by demethylation of O-6-methylguanine-DNA methyltransferase (MGMT).
  • MGMT O-6-methylguanine-DNA methyltransferase
  • PARP Poly(ADP)-ribose polymerase
  • PARP family members At present, at least 17 members of the PARP family have been found, and all PARP family proteins have similar catalytic domain sequences. In fact, only a small part of the PARP family members synthesize poly(adenosine diphosphate-ribose) (PAR) with Nicotinamide adenine dinucleotide (NAD + ) as a substrate. It is transferred to a receptor protein to regulate the function of related proteins.
  • PARPAR poly(adenosine diphosphate-ribose)
  • NAD + Nicotinamide adenine dinucleotide
  • PARP1 In the PARP family, only PARP1/2 can be activated by DNA single-strand breaks and mediate poly ADP ribosylation, and participate in the repair of DNA damage via the base excision repair pathway.
  • Human PARP1 is a polypeptide chain with a molecular weight of 113 kDa and contains three functional domains.
  • the DNA binding domain (DBD) at the N-terminus contains two zinc finger fingers that recognize DNA breaks. The first zinc finger recognizes DNA fragmentation and activation of PARP1 activity, its damage will completely inactivate PARP1, and the second zinc finger is only involved in the recognition of DNA single-strand breaks.
  • PARP1 In the middle part of the Automodification domain (AMD), PARP1 binds to the poly ADP ribose group through this region, and undergoes self-poly ADP ribosylation to regulate its interaction with DNA or protein.
  • the C-terminal catalytic domain the most strictly conserved part of PARP1, catalyzes the synthesis of PAR and the domain that phosphatizes the target protein into ADP.
  • Human PARP2 is a polypeptide chain with a molecular weight of 62 kDa. Unlike PARP1, the N-terminus of PARP2 does not contain a zinc finger structure, so the effectiveness of PARP2 for SSB recognition is reduced, which in turn identifies gaps in the DNA damage chain due to nucleotide loss.
  • PARP2 is substantially similar to the catalytic domain of PARP1, with 69% homology, but the subtle differences in structure reflect differences in their function.
  • PARP1 inhibitors were used as chemosensitizers to kill tumor cells in combination with radiotherapy and chemotherapy.
  • PARP1 inhibitors specifically inhibit/kill breast cancer genes BRCA1 and BRCA2 deletion cells.
  • DNA double-stranded repairs include HR and NHEJ, but NHEJ may have errors that lead to genomic instability.
  • HR is mainly mediated by two key genes, BRCA1 and BRCA2.
  • BRCA-deficient cells repairing DNA double-strand breaks requires the NHEJ pathway, which leads to the onset of tumors.
  • BRCA1/2 deficiency is clinically prone to breast cancer, ovarian cancer, prostate cancer, pancreatic cancer, etc. This HR pathway inactivated tumor is called "BRCAness".
  • the structure-activity relationship of PARP inhibitors mainly has the following aspects: maintenance activity and binding force must contain at least one hydrogen free formamide structure, which can be fused to a bicyclic structure or a "pseudobicyclic" system, and simultaneously becomes a hydrogen bond. Receptors and hydrogen bond donors; bulky hydrophobic groups for altering activity and physicochemical properties, and the like.
  • AZD2281 has been approved for marketing.
  • most PARP1/2 inhibitors have the disadvantage of low bioavailability and poor subtype selectivity.
  • An object of the present invention is to provide a 2-substituted benzimidazole-4-carboxamide compound, a preparation method and application thereof.
  • R is hydrogen, a linear or branched alkyl group of C1-C4;
  • R 1 is hydrogen, a substituted or unsubstituted C1-C4 linear or branched alkyl group, or a substituted or unsubstituted C3-C6 cycloalkyl group;
  • R 2 is a free, halogen, substituted or unsubstituted C1-C4 linear or branched alkyl group, or a substituted or unsubstituted C3-C6 cycloalkyl group;
  • Y 1 , Y 2 are independently selected from substituted or unsubstituted methylene, substituted or unsubstituted ethylene, substituted or unsubstituted propylene or substituted or unsubstituted butylene;
  • X is CR 6 or N, wherein R 6 is hydrogen, a substituted or unsubstituted C1-C4 linear or branched alkyl group, a substituted or unsubstituted C1-C4 linear or branched alkoxy group, halogen or a substituted or unsubstituted C3-C6 cycloalkyl group;
  • each substitution independently refers to having from 1 to 3 substituents selected from the group consisting of hydroxy, halo, C1-C6 straight or branched alkyl, C1-C4 straight or branched alkoxy, C3- a cycloalkyl group, a C6-C10 aryl group or a carboxyl group of C6; the C1-C6 linear or branched alkyl group in the substituent may be further amino, hydroxy, methoxy, ethoxy, propoxy or iso Propyl, -CO 2 Me, -CO 2 Et, -CO 2 Pr or -CO 2 Pr-i are substituted.
  • the invention provides a compound of formula I, an R-isomer, an S-isomer, or a pharmaceutically acceptable salt thereof:
  • R 1 is hydrogen, a substituted or unsubstituted C1-C4 linear or branched alkyl group, or a substituted or unsubstituted C3-C6 cycloalkyl group;
  • R 2 is a free, halogen, substituted or unsubstituted C1-C4 linear or branched alkyl group, or a substituted or unsubstituted C3-C6 cycloalkyl group;
  • Y 1 , Y 2 are independently selected from substituted or unsubstituted methylene, substituted or unsubstituted ethylene, substituted or unsubstituted propylene or substituted or unsubstituted butylene;
  • X is CR 6 or N, wherein R 6 is hydrogen, a substituted or unsubstituted C1-C4 linear or branched alkyl group or a substituted or unsubstituted C3-C6 cycloalkyl group;
  • each substitution independently refers to having from 1 to 3 substituents selected from the group consisting of hydroxy, halo, C1-C6 straight or branched alkyl, C1-C4 straight or branched alkoxy, C3- a cycloalkyl group of C6, a C6-C10 aryl group, or a carboxyl group.
  • R is hydrogen, methyl, ethyl.
  • R 1 is hydrogen, a C1-C4 linear or branched alkyl group, a halogen-substituted C1-C4 linear or branched alkyl group or a C3-C6 cycloalkyl group.
  • R 1 is hydrogen, methyl, ethyl, monofluoromethyl, trifluoromethyl or difluoromethyl.
  • R 2 is hydrogen, fluorine, chlorine, bromine, a C1-C4 linear or branched alkyl group, or a C3-C6 cycloalkyl group.
  • R 2 is none, fluorine, chlorine, methyl, ethyl.
  • R 3 is hydrogen, methyl, ethyl, acetyl, cyclopropanoyl, benzoyl, methylsulfonyl, -COCH 2 OCH 2 CH 2 CH 3 .
  • X is CR 6 or N, wherein R 6 is hydrogen, a C1-C4 linear or branched alkyl group or a C3-C6 cycloalkyl group.
  • X is CH, C(CH 3 ), C(CH 2 CH 3 ) or N.
  • Y 1 is a substituted or unsubstituted methylene group, a Y 2 -substituted or unsubstituted ethylene group; or Y 1 is a substituted or unsubstituted ethylene group, and Y 2 is a substituted or unsubstituted group.
  • Methylene of which
  • Each of the substituents independently means having 1-2 substituents selected from the group consisting of hydroxy, halogen, C1-C4 straight or branched alkyl, C1-C4 straight or branched alkoxy, C3-C6 a cycloalkyl group; wherein the C1-C4 straight or branched alkyl group may be further amino, hydroxy, methoxy, ethoxy, propoxy, isopropoxy, -CO 2 Me, -CO 2 Et , -CO 2 Pr or -CO 2 Pr-i substituted.
  • Y 1 is a substituted or unsubstituted methylene group, a Y 2 -substituted or unsubstituted ethylene group; or Y 1 is a substituted or unsubstituted ethylene group, and Y 2 is a substituted or unsubstituted group.
  • a methylene group wherein the substituent independently means having 1 to 2 substituents selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tertiary.
  • the compound is any one of the compounds I-1 to I-33, I-36 to I-45 prepared in the examples.
  • a process for the preparation of a compound according to the first aspect which comprises the step of obtaining a compound of the formula Ia by aminolysis of a compound of the formula IIa:
  • R, R 1 , R 2 , Y 1 , Y 2 , R 3 , X are as defined in the first aspect, and R 7 is selected from a linear or branched alkyl group of C1-C4.
  • the above method comprises the steps of: subjecting the compound of Formula II to aminolysis to give a compound of Formula I:
  • R 1 , R 2 , Y 1 , Y 2 , R 3 , X are as defined in the first aspect.
  • a third aspect of the invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising the compound of the first aspect, the R-isomer, the S-isomer, or a pharmaceutically acceptable salt thereof;
  • a pharmaceutically acceptable carrier is selected from:
  • a fourth aspect of the invention provides the use of the compound of the first aspect or the pharmaceutical composition of the third aspect for the preparation of:
  • a drug for preventing and/or treating a disease associated with PARP is a drug for preventing and/or treating a disease associated with PARP.
  • R 1 , R 2 , Y 1 , Y 2 , R 3 , X are as defined in the first aspect, and R 7 is selected from C1-C4 straight or branched alkyl groups, preferably from methyl, ethyl and undertiary Butyl.
  • the compound of Formula IIa is as shown in Formula II:
  • R 1 , R 2 , Y 1 , Y 2 , R 3 , and X are as described in the first aspect.
  • the inventors of the present application have extensively and intensively studied to design and synthesize a class of benzimidazole-4-carboxamide PARP1/2 inhibitors containing sulfur atoms for the first time.
  • the compounds of the present invention have a well-defined structure-activity relationship and exhibit excellent anti-tumor activity in vitro and in vivo.
  • This new type of PARP1/2 inhibitor is expected to become a new anti-tumor drug.
  • the present invention has been completed on this basis.
  • alkyl means a saturated linear or branched hydrocarbon moiety, such as -CH 3 or -CH (CH 3) 2.
  • alkoxy denotes an -O-(alkyl) group.
  • cycloalkyl denotes a saturated cyclic hydrocarbyl moiety such as cyclopropyl, cyclohexyl.
  • aryl refers to a hydrocarbyl moiety containing one or more aromatic rings. Examples of the aryl moiety include phenyl (Ph), naphthyl and the like.
  • alkyl, alkoxy, cycloalkyl and aryl groups described herein include both substituted and unsubstituted moieties.
  • Possible substituents on alkyl, alkoxy, cycloalkyl and aryl groups include, but are not limited to, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C10 cycloalkyl, C1 -C20 heterocycloalkyl, C1-C10 alkoxy, C6-C10 aryl, amino, C1-C10 alkylamino, C1-C20 dialkylamino, cyano, nitro and carboxy.
  • the compound of the formula Ia provided by the present invention has the following structure:
  • R, R 1 , R 2 , Y 1 , Y 2 , R 3 and X are as defined above.
  • the compounds of the invention have asymmetric centers, chiral axes and chiral planes, and can be racemic, R- The form of the isomer or S-isomer is present. Those skilled in the art will be able to resolve the R-isomer and/or the S-isomer from the racemate by conventional techniques.
  • a pharmaceutically acceptable salt of a compound of formula Ia in particular a compound of formula Ia, which is reacted with an inorganic or organic acid to form a conventional pharmaceutically acceptable salt.
  • a conventional pharmaceutically acceptable salt can be prepared by reacting a compound of the formula Ia with an inorganic or organic acid, including hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, aminosulfonic acid, phosphoric acid, and the like, and The organic acid includes citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, naphthalenesulfonic acid, ethanesulfonic acid, naphthalene disulfonic acid, maleic acid, malic acid, Malonic acid, fumaric acid, succinic acid, propionic acid, oxalic acid, trifluor
  • the invention provides a compound of formula I, an R-isomer, an S-isomer, or a pharmaceutically acceptable salt thereof:
  • R 1 , R 2 , Y 1 , Y 2 , R 3 and X are as indicated above.
  • the o-phenylenedicarboxylate compound 1 and the aldehyde derivative 2 are condensed and cyclized in the presence of sodium hydrogen sulfite or other Lewis acid and oxidized to form the compound 3; the compound 3 is removed in the presence of an acid such as trifluoroacetic acid.
  • the Boc protecting group generates the compound 4, and the compound 4 is aminolyzed by the aminolysis reagent to give the compound I-1, I-4, I-6 to I-8, I-15, I-17 to I-29, I-36 to I. -40.
  • Compound 3 is firstly aminolyzed by an aminolysis reagent to form compound 5, and compound 5 is obtained by removing the Boc protecting group under the action of an acid such as hydrochloric acid or trifluoroacetic acid to obtain I-1, I-4, I-6 to I-8, I. -15, I-17 to I-29, I-36 to I-40.
  • an acid such as hydrochloric acid or trifluoroacetic acid
  • the o-phenylenediaminecarboxylate compound 1 is reacted with an aminolysis reagent to obtain a compound 6, which is condensed and cyclized in the presence of sodium hydrogen sulfite or other Lewis acid to form a compound 5, and the compound 5 is passed through a trifluorocarbon.
  • the Boc protecting group is deactivated by the action of an acid such as acetic acid or hydrochloric acid to give the compound I-1, I-4, I-6 to I-8, I-15, I-17 to I-29, and I-36 to I-40.
  • Compound 3 and R 1 L or sodium difluorochloroacetate form compound 7 under the action of a base, and compound 7 is decarboxylated to form compound 8 under the action of an acid such as trifluoroacetic acid or hydrochloric acid, and compound 8 is under the action of an aminolysis reagent.
  • Compounds I-30 to I-33 and I-41 to 43 are produced, wherein the L is a halogen.
  • Compound 5 and R 1 L or sodium difluorochloroacetate form compound 9 under the action of a base, and compound 9 is decarboxylated to form compounds I-30 to I-33 and I- under the action of an acid such as trifluoroacetic acid or hydrochloric acid. 41 to 43, wherein L is a halogen.
  • R, R 1 , R 2 , R 3 , X, Y 1 and Y 2 are as defined above; the amino reagent is selected from the group consisting of ammonia methanol solution and ammonia ethanol solution.
  • intermediate 1 is commercially available or is prepared as follows:
  • the ruthenium compound 11 is nitrated to form a nitro compound, and the nitro compound is oxidized by Baeyer-Villiger in the presence of a peroxide such as hydrogen peroxide and hydrolyzed and opened under basic conditions to form a carboxylic acid compound, and the carboxylic acid compound is esterified under acid catalysis. Forming an ester, which is reduced by palladium hydrocarbonation or other reducing agent such as iron powder, zinc powder or the like to form compound 1;
  • the aldehyde intermediate 2 can be obtained commercially or by the following procedure:
  • the invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising an active ingredient in a safe and effective amount, together with a pharmaceutically acceptable carrier.
  • the "active ingredient” as used in the present invention means a 2-substituted benzimidazole-4-carboxamide compound represented by the formula I according to the present invention.
  • the 2-substituted benzimidazole-4-carboxamides and pharmaceutical compositions of the present invention can be used as PARP1 inhibitors or PARP2 inhibitors; can be used for the preparation of prophylactic and/or therapeutic agents with poly ADP-ribose polymerase A drug for a PARP-related disease; a drug which can be used for the preparation of a prophylactic and/or therapeutic tumor; and an anti-inflammatory drug.
  • the disease associated with poly ADP-ribose polymerase PARP is tumor, inflammation, and ischemic-reperfusion complicated diseases such as cardiovascular disease, diabetes, rheumatoid arthritis, endotoxin Shock, stroke, etc.
  • the tumor is ovarian cancer, breast cancer, gastric cancer, prostate cancer, lung cancer, advanced solid tumor, glioma, melanoma, colon cancer, pancreatic cancer, recurrent hematological cancer, Ewing's sarcoma, Pancreatic cancer, endometrial cancer, etc.
  • the tumor is a tumor defective in homologous recombination repair, that is, a tumor with BRCA1 or BRCA2 deletion or mutation, such as ovarian cancer, breast cancer, prostate cancer, gastric cancer, pancreatic cancer, cervical cancer, glial Tumor, Ewing's sarcoma, etc.
  • a tumor with BRCA1 or BRCA2 deletion or mutation such as ovarian cancer, breast cancer, prostate cancer, gastric cancer, pancreatic cancer, cervical cancer, glial Tumor, Ewing's sarcoma, etc.
  • the pharmaceutical compositions contain from 1 to 2000 mg of active ingredient per dose, more preferably from 10 to 200 mg of active ingredient per dose.
  • the "one dose” is a tablet.
  • “Pharmaceutically acceptable carrier” means: one or more compatible solid or liquid fillers or gel materials which are suitable for human use and which must be of sufficient purity and of sufficiently low toxicity.
  • “compatibility” it is meant herein that the components of the composition are capable of intermingling with the active ingredients of the present invention and with respect to each other without significantly reducing the efficacy of the active ingredients.
  • pharmaceutically acceptable carriers are cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid).
  • magnesium stearate magnesium stearate
  • calcium sulfate vegetable oil (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyol (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifier Wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.
  • the 2-substituted benzimidazole-4-carboxamide compound of the formula I of the present invention can form a complex with a macromolecular compound or a polymer by non-bonding.
  • the 2-substituted benzimidazole-4-carboxamide compound of the formula I of the present invention can be linked as a small molecule to a macromolecular compound or a polymer by a chemical bond.
  • the macromolecular compound may be a biological macromolecule such as a polysaccharide, a protein, a nucleic acid, a polypeptide, or the like.
  • the administration form of the active ingredient or the pharmaceutical composition of the present invention is not particularly limited, and representative administration forms include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous) and the like.
  • Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.
  • the active ingredient is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or mixed with: (a) a filler or compatibilizer, for example, Starch, lactose, sucrose, glucose, sweet (l) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerin; (d) collapse Decomposing agents, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) a slow solvent such as paraffin; (f) an absorption accelerator, for example, a quaternary amine a compound; (g) a wetting agent such as cetyl alcohol and glyceryl monostearate; (h) an adsorbent such as kaolin; and (
  • the solid dosage forms can also be prepared with coatings and shell materials, such as casings and other materials known in the art. They may contain opacifying agents and the release of the active ingredient in such compositions may be released in a portion of the digestive tract in a delayed manner. Examples of embedding components that can be employed are polymeric and waxy materials.
  • Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or elixirs.
  • the liquid dosage form may contain inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or a mixture of these substances.
  • the compositions may contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and perfumes.
  • the suspension may contain suspending agents, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these and the like.
  • suspending agents for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these and the like.
  • compositions for parenteral injection may comprise a physiologically acceptable sterile aqueous or nonaqueous solution, dispersion, suspension or emulsion, and a sterile powder for reconstitution into a sterile injectable solution or dispersion.
  • Suitable aqueous and nonaqueous vehicles, diluents, solvents or vehicles include water, ethanol, polyols, and suitable mixtures thereof.
  • the compounds of the invention may be administered alone or in combination with other therapeutic agents, such as chemotherapeutic agents.
  • a safe and effective amount of a compound of the invention is administered to a mammal (e.g., a human) in need of treatment wherein the dosage is a pharmaceutically effective effective dosage, for a 60 kg body weight
  • the dose to be administered is usually from 1 to 2000 mg, preferably from 20 to 500 mg.
  • specific doses should also consider factors such as the route of administration, the health of the patient, etc., which are within the skill of the skilled physician.
  • 1 H NMR was recorded by a Varian Mercury-300 or Varian Mercury-400 nuclear magnetic resonance apparatus
  • 13 C NMR was recorded by a Varian Mercury-400 or Varian Mercury-500 or Varian Mercury-600 nuclear magnetic resonance apparatus, chemistry.
  • the displacement is expressed in ⁇ (ppm);
  • the mass spectrum is recorded by Finnigan/MAT-95 (EI) and Finnigan LCQ/DECA and Micromass Ultra Q-TOF (ESI) mass spectrometer; the separation silica gel is 200-300 mesh.
  • 4,5,6,7-Tetrahydrothiophene [3,2-c]pyridine hydrochloride (10 g, 56.9 mmol) was weighed in a reaction flask (500 mL) and dichloromethane (300 mL). Triethylamine (8.7 mL) was added under ice-cooling, and a solution of di-tert-butyl dicarbonate (13.7 g, 62.8 mmol) in dichloromethane (20 mL). After the completion of the dropwise addition, the reaction was followed for 1 hour, and the reaction of the starting material was completed.
  • 6-fluoro-2-(4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole-4 was dissolved at 0 °C - acetyl chloride (57 mg, 0.72 mmol) was slowly added dropwise to a solution of methyl formate (198 mg, 0.60 mmol) and triethylamine (121 mg, 1.20 mmol) in anhydrous DMF (4 mL). In a mixture of ice and water, a white solid was precipitated, filtered, and dried in vacuo to give 197 mg.
  • N-(2-(thien-3-yl)ethyl)acetamide (497 mg, 2.9 mmol) was dissolved in 3 mL of phosphorus oxychloride, heated to 60 ° C, reacted for two hours, cooled, and the reaction solution was poured into The mixture was neutralized with a saturated Na 2 CO 3 solution to dryness, and extracted with ethyl acetate (50 mL ⁇ 3). The organic layer was washed successively with water and brine, and the organic layer was dried. concentrated under pressure to give a yellow oil.
  • N-(2-(thiophen-3-yl)ethyl)propanamide (16-2) (543 mg, 3.0 mmol) was used as the starting material to obtain 529 mg of colorless oil.
  • N-(2-(thien-3-yl)ethyl)butanamide (16-3) (585 mg, 3.0) Methyl) 532 mg of a colorless oil was obtained from the crude material.
  • N-(2-(thiophen-3-yl)ethyl)pentanamide (16-4) (627 mg, 3.0 mmol)
  • N-(2-(thiophen-3-yl)ethyl)cyclopropylformamide (16-7) (627 mg, 3.0 mmol)
  • the separation conditions are as in S-4-13.
  • Di-tert-butyl dicarbonate (1.78 g, 10.0 mmol) was slowly added to the organic layer at 0 ° C, and the mixture was stirred at room temperature for 1 hour, and most of the solvent was evaporated under reduced pressure. The yield was 55%.
  • N-tert-butoxycarbonyl-4-piperidone (22) (9.95 g, 50.0 mmol) in cyclohexane (25 mL) was added pyrrolidine (4.35 mL, 52.0 mmol) and p-toluene
  • the acid 48 mg, 2.5 mmol was refluxed with a water separator for 50 minutes, cooled to room temperature, evaporated to dryness under reduced pressure, and then redissolved in 15 mL of anhydrous methanol, and then 1.60 g of sulfur powder was added to the mixture, and stirred at room temperature for 20 minutes.
  • the free I-30 (100 mg) was dissolved in a mixed solvent of dichloromethane (100 ml) and methanol (10 ml), and then slowly stirred for 30 minutes HCl (g) in an ice bath, then stirred at room temperature overnight, concentrated to remove most Solvent, filter and dry to give the I-30 hydrochloride.
  • Step 1 ((5-Bromothiophen-3-yl)methyl)(methyl)carbamic acid tert-butyl ester:
  • Step 2 ((5-Formylthiophen-2-yl)methyl)(methyl)carbamic acid tert-butyl ester:
  • step b) The product from step b) (156 mg, 0.42 mmol) was added to tetrahydrofuran (10 ml), and then di-tert-butyl dicarbonate (109 mg, 0.5 mmol) was added to react under reflux until the main material disappeared.
  • Step 6 7-(2-(tert-Butoxycarbonylamino)propyl-2-yl)-2-(4-carbamoyl-6-fluoro-1H-benzo[d]imidazol-2-yl)- 4,5-dihydrothiophene [2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (36-6)
  • Step 1 2-(4-carbamoyl-6-fluoro-1-methyl-1H-benzo[d]imidazol-2-yl)-7-(2-methoxyisopropyl-2-yl) -4,5-dihydrothieno[2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (41-1) and 2-(6-fluoro-1-methyl-4(methylamine) Formyl)-1H-benzo[d]imidazol-2-yl)-7-(2-methoxyisopropyl-2-yl)-4,5-dihydrothieno[2,3-c]pyridine -6(7H)-tert-butyl carboxylic acid (41-2)
  • Step 1 2-(4-Carboxamido-1-difluoromethyl)-6-fluoro-1H-benzo[d]imidazol-2-yl)-7-(2-methoxyisopropyl-2 -yl)-4,5-dihydrothieno[2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (43-1)
  • the principle is to coat the substrate histone in an adsorption 96-well plate, add PARP1/2 recombinase, substrate NAD + , activated DNA to enzymatically react PARP1/2, PARP enzyme can catalyze the cleavage of substrate DAD + amide bond to form nicotinamide and ADP ribose, and synthesize poly(ADP) ribose (PAR) on target protein (Histone) with ADP ribose as substrate, then add anti-PAR (anti-PAR) antibody, detect The intensity of the product PAR on the histones coated on the 96-well plate reflects the PARP enzyme activity.
  • Histone is an important substrate recognized by PARP.
  • Histone (1 ng/mL) was coated with 96-well microtiter plate with potassium-free PBS (10 mM sodium phosphate buffer, 150 mM NaCl, pH 7.2-7.4), and coated overnight at 37 ° C; discarded in the well liquid.
  • the plate was washed 5 times with 120 ⁇ L/well of T-PBS (PBS containing 0.1% Tween-20), and dried in an oven at 37 °C.
  • OPD O-Phenylenediamine Dihydrochloride
  • the degree of inhibition of PARP1 enzyme activity by the drug was calculated according to the following formula:
  • Inhibition rate (%) [(OD positive control well- OD negative control well ) - (OD administration well - OD negative control well )] / (OD positive control well - OD negative control well ) ⁇ 100%
  • the experimental procedure was the same as the inhibition of the PARP1 enzyme activity at the molecular level.
  • the detailed data of the added enzyme and substrate were: addition of NAD + (final concentration 80 ⁇ M), DNA (100 ng/well), and PARP 2 (10 ng/well).
  • CCK8 Cell Counting Kit-8
  • CCK-8 reagent contains WST-8: chemical name: 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfonate Acid benzene)-2H-tetrazole monosodium salt, which is reduced by dehydrogenase in cell mitochondria under the action of electron carrier 1-methoxy-5-methylphenazine dimethyl sulfate (1-Methoxy PMS) It is a yellow formazan product (Formazan) with high water solubility. The amount of formazan produced is proportional to the number of living cells. The more the cells proliferate, the darker the color; the greater the cytotoxicity, the lighter the color. For the same cells, the color depth and the number of cells are linear. The light absorption value was measured by a microplate reader at a wavelength of 450 nm, which indirectly reflected the number of living cells, thereby calculating the cell inhibition rate.
  • the specific method is as follows: the cells in the logarithmic growth phase are inoculated into the 96-well culture plate at a density of 2000 cells/well of V79 cells and 4000 cells/well of VC8 cells, 90 ⁇ L per well, and cultured overnight, and then added at different concentrations (final concentration from 10 ⁇ L of the drug was started at 10 ⁇ M, and 5 ⁇ L of 6 gradients were applied for 72 h. Three wells were set for each concentration, and the corresponding concentrations of physiological saline vehicle control and cell-free zero-adjusted wells were set. After the end of the action, 10 ⁇ L of CCK8 reagent was added, and the culture was continued at 37 ° C for 4 hours, and the OD450 value was measured by a microplate reader.
  • Inhibition rate (%) (OD control well- OD administration well ) / OD control well ⁇ 100%
  • the drug concentration at the 50% inhibition rate that is, the IC 50 value, was calculated according to the Logit method.
  • the inhibitory effect of 3 compounds on PARP1 in Table 2 was comparable to that of the positive compound AZD2281, IC 50 (nM): I-37 (18.03 nM), I-39 (33.50 nM), I-44 (17.78 nM).
  • the intensity of action on PARP2 was weaker than that of the positive control AZD2281, but some compounds exerted stronger effects on PARP2 than PARP1, indicating selectivity for PARP2.
  • compound I-35 adopts a ring-opening strategy, and its inhibitory activity against PARP1 is also decreased, but the activity of PARP2 is enhanced.
  • Animals BALB/cA nude mice, female, 4-5 weeks old, weighing 19 ⁇ 2 g, provided by Shanghai Institute of Materia Medica, Chinese Academy of Sciences, production license number: SCXK (Shanghai) 2013-001. Use certificate number: SYXK (Shanghai) 2013-0049.
  • Cell line Human breast cancer MDA-MB-436 cell line was inoculated subcutaneously into the right axilla of nude mice, and the amount of cells inoculated was 5 ⁇ 10 6 /piece. After the transplanted tumor was formed, it was used in nude mice for 2 generations.
  • the tumor tissue in the vigorous growth period was cut into 1.5 mm 3 and inoculated subcutaneously in the right axilla of nude mice under aseptic conditions.
  • the diameter of the transplanted tumor was measured with a vernier caliper in nude mice.
  • the animals were randomly divided into groups.
  • the I-30 monohydrochloride 100 mg/kg and 20 mg/kg groups were orally administered once a day for 21 consecutive days.
  • the positive control drug AZD2281 30 mg/kg was orally administered once a day for 21 days.
  • the solvent control was given an equal amount of water for injection.
  • the diameter of the transplanted tumor was measured twice a week during the entire experiment, and the body weight of the mice was weighed.
  • the formula for calculating tumor volume (TV) is:

Abstract

The present invention relates to a 2-substituted benzimidazole-4-carboxamide compound and the preparation method and use thereof. The structure of the 2-substituted benzimidazole-4-carboxamide compound of the present invention is as shown in the general formula Ia, wherein the definitions of R, R1, R2, Y1, Y2, R3 and X are as shown in the claims and the description. Also disclosed is a pharmaceutical composition, comprising a 2-substituted benzimidazole-4-carboxamide compound or a pharmaceutically acceptable salt thereof. The 2-substituted benzimidazole-4-carboxamide compound and the pharmaceutical composition of the present invention can be used in the preparation of drugs for the treatment of diseases associated with polyadenosine diphosphate ribose polymerases, such as malignant tumors.

Description

2-取代苯并咪唑-4-甲酰胺类化合物及制备方法和应用2-substituted benzimidazole-4-carboxamide compound, preparation method and application thereof 技术领域Technical field
本发明属于药物化学和药物治疗学领域。具体涉及一类2-(噻吩或噻唑并四氢吡啶-2-基)-1H-苯并咪唑-4-甲酰胺类化合物或其药学上可接受的盐，其制备方法及它们在制备治疗与聚腺苷二磷酸核糖聚合酶(PARP)相关的疾病如恶性肿瘤的药物中的应用。The invention belongs to the fields of medicinal chemistry and pharmacotherapy. Specifically relates to a class of 2-(thiophene or thiazolotetrahydropyridin-2-yl)-1H-benzimidazole-4-carboxamides or pharmaceutically acceptable salts thereof, processes for their preparation and their preparation and treatment The use of poly ADP-ribose polymerase (PARP)-related diseases such as malignant tumors.
背景技术Background technique
人体内每天大约有1013个细胞受到数万次DNA损伤，暴露于恶劣的环境中(例如：紫外辐射、离子辐射等)、正常的细胞代谢副产物、细胞毒药物都会导致DNA损伤。这些损伤包括碱基修饰、单链断裂(Single strand breaks,SSB)、双链断裂(Double strand breaks,DSB)、交叉连接等。DNA损伤严重影响基因的复制和转录，甚至造成基因组畸变和肿瘤的产生。为了维持基因组的完整稳定性，人体系统存在多个途径检测DNA损伤并每时每刻对DNA损伤进行修复。主要的DNA修复途径包括碱基切除修复(Base-excision repair,BER)、核苷酸切除修复(Nucleotide-excision repair,NER)、错配修复(Mismatch repair,MMR)、同源重组(Homologous recombination,HR)及非同源末端连接(Nonhomologous end joining,NHEJ)等。其中BER、NER、MMR是主要的DNA单链断裂(SSB)修复途径，HR和NHEJ是主要的DNA双链断裂(DSB)修复途径，HR是可靠准确的修复途径，而NHEJ并不稳定，容易出错。另外有些DNA损伤可以被相应的酶直接修复，例如鸟嘌呤甲基化可被O-6-甲基鸟嘌呤-DNA甲基转移酶(MGMT)去甲基化修复。癌症患者在使用细胞毒药物后，肿瘤细胞DNA损伤修复增加，造成细胞凋亡耐受，是肿瘤耐药性产生的重要原因之一。About 10 13 cells in the human body are damaged by tens of thousands of DNA every day. Exposure to harsh environments (such as ultraviolet radiation, ionizing radiation, etc.), normal cellular metabolic by-products, and cytotoxic drugs can cause DNA damage. These damages include base modifications, single strand breaks (SSB), double strand breaks (DSB), cross-linking, and the like. DNA damage seriously affects the replication and transcription of genes, and even causes genomic aberrations and tumors. In order to maintain the complete stability of the genome, there are multiple pathways in the human system to detect DNA damage and repair DNA damage at all times. The main DNA repair pathways include base-excision repair (BER), Nucleotide-excision repair (NER), mismatch repair (MMR), and homologous recombination (Homologous recombination, HR) and Nonhomologous end joining (NHEJ) and the like. Among them, BER, NER and MMR are the main DNA single-strand break (SSB) repair pathways. HR and NHEJ are the main DNA double-strand break (DSB) repair pathways. HR is a reliable and accurate repair pathway, while NHEJ is unstable and easy. An error occurred. In addition, some DNA damage can be directly repaired by the corresponding enzymes. For example, guanine methylation can be repaired by demethylation of O-6-methylguanine-DNA methyltransferase (MGMT). Cancer patients use cytotoxic drugs, tumor cell DNA damage repair increased, resulting in apoptosis tolerance, is one of the important reasons for tumor resistance.
聚腺苷二磷酸核糖聚合酶(Poly(ADP)-ribose polymerase，PARP)是一类存在于多数真核细胞中的细胞核酶。目前，至少已有17个PARP家族成员被发现，所有PARP家族蛋白都有相似的催化结构域序列。事实上，PARP家族成员中只有少部分是以烟酰胺腺嘌呤二核苷酸(Nicotinamide adenine dinucleotide,NAD+)为底物合成聚腺苷二磷酸核糖(Poly(adenosine diphosphate-ribose)，PAR)，并将其转移至受体蛋白，调控相关蛋白的功能。在PARP家族中，只有PARP1/2能够被DNA单链断裂激活并介导聚ADP核糖化，经由碱基切除修复途径参与DNA损伤的修复。人类PARP1是一条分子量为113kDa的多肽链，包含3个功能性结构域。位于N-端的DNA结合结构域(DNA binding domain,DBD)，含有两个锌指结构(Zinc finger)，识别DNA断裂。第1个锌指识别DNA断裂和PARP1活性的激活，它的损伤将彻底使PARP1丧失活性，第2个锌指只参与DNA单链断裂的识别。中间部分的自修饰结构域(Automodification domain,AMD)，PARP1通过该区与聚ADP核糖基结合，发生自身聚ADP核糖化，调节自身与DNA或蛋白的相互作用。C-端的催化结构域(Catalytic domain)，是PARP1最严格保守的部分，催化合成PAR以及将目标蛋白聚ADP核糖化的结构域。人类PARP2是一条分子量为 62kDa的多肽链。与PARP1不同，PARP2的N-端不含锌指结构，因此PARP2对SSB识别的有效性降低，转而识别DNA损伤链上由于核苷酸丢失而形成的空隙。PARP2与PARP1的催化结构域基本相似，具有69％的同源性，但结构的细微差别还是反应出它们功能的差异。为了对抗肿瘤耐药性的产生以及降低细胞坏死而产生的炎症反应，早期，PARP1抑制剂被用作化学增敏剂，与放、化疗联合作用杀灭肿瘤细胞。Poly(ADP)-ribose polymerase (PARP) is a class of nuclear enzymes found in most eukaryotic cells. At present, at least 17 members of the PARP family have been found, and all PARP family proteins have similar catalytic domain sequences. In fact, only a small part of the PARP family members synthesize poly(adenosine diphosphate-ribose) (PAR) with Nicotinamide adenine dinucleotide (NAD + ) as a substrate. It is transferred to a receptor protein to regulate the function of related proteins. In the PARP family, only PARP1/2 can be activated by DNA single-strand breaks and mediate poly ADP ribosylation, and participate in the repair of DNA damage via the base excision repair pathway. Human PARP1 is a polypeptide chain with a molecular weight of 113 kDa and contains three functional domains. The DNA binding domain (DBD) at the N-terminus contains two zinc finger fingers that recognize DNA breaks. The first zinc finger recognizes DNA fragmentation and activation of PARP1 activity, its damage will completely inactivate PARP1, and the second zinc finger is only involved in the recognition of DNA single-strand breaks. In the middle part of the Automodification domain (AMD), PARP1 binds to the poly ADP ribose group through this region, and undergoes self-poly ADP ribosylation to regulate its interaction with DNA or protein. The C-terminal catalytic domain, the most strictly conserved part of PARP1, catalyzes the synthesis of PAR and the domain that phosphatizes the target protein into ADP. Human PARP2 is a polypeptide chain with a molecular weight of 62 kDa. Unlike PARP1, the N-terminus of PARP2 does not contain a zinc finger structure, so the effectiveness of PARP2 for SSB recognition is reduced, which in turn identifies gaps in the DNA damage chain due to nucleotide loss. PARP2 is substantially similar to the catalytic domain of PARP1, with 69% homology, but the subtle differences in structure reflect differences in their function. In order to combat the development of tumor resistance and reduce the inflammatory response caused by cell necrosis, early, PARP1 inhibitors were used as chemosensitizers to kill tumor cells in combination with radiotherapy and chemotherapy.
直至2005年，两个不同的课题组同时在Nature发表了PARP研究领域的一个重大突破：PARP1抑制剂特异性抑制/杀伤乳腺癌基因BRCA1和BRCA2缺失的细胞。如前所述，DNA双链修复包括HR和NHEJ，但NHEJ有可能会出现错误，导致基因组的不稳定。HR主要由BRCA1和BRCA2两个关键基因介导，在BRCA缺失的细胞中，修复DNA双链断裂需要通过NHEJ途径，因而导致容易发生肿瘤。BRCA1/2缺失临床上易表现为乳腺癌，卵巢癌，前列腺癌，胰腺癌等，这种HR途径失活的肿瘤被称作“BRCAness”。在HR缺陷的细胞中，抑制PARP1将导致DNA单链断裂蓄积增加，转化为致死性DSBs。致死性的DSBs导致染色体畸变以及基因组的不稳定性，产生细胞死亡。即，一个基因功能的缺失将导致细胞敏感(如PARP1或者BRCA1/2的缺失)，但是两个基因功能的缺失却是致命的(如PARP1和BRCA1/2同时缺失)。BRCAness与PARP1/2抑制共同杀伤细胞的这种现象称为协同致死(Synthetic lethality)。协同致死理论为恶性肿瘤的治疗提供了新策略和新思路，自此，PARP1抑制剂的研究进入了一个全新的时代。Until 2005, two different research groups simultaneously published a major breakthrough in the field of PARP research in Nature: PARP1 inhibitors specifically inhibit/kill breast cancer genes BRCA1 and BRCA2 deletion cells. As mentioned earlier, DNA double-stranded repairs include HR and NHEJ, but NHEJ may have errors that lead to genomic instability. HR is mainly mediated by two key genes, BRCA1 and BRCA2. In BRCA-deficient cells, repairing DNA double-strand breaks requires the NHEJ pathway, which leads to the onset of tumors. BRCA1/2 deficiency is clinically prone to breast cancer, ovarian cancer, prostate cancer, pancreatic cancer, etc. This HR pathway inactivated tumor is called "BRCAness". In HR-deficient cells, inhibition of PARP1 results in increased accumulation of DNA single-strand breaks and conversion to lethal DSBs. Lethal DSBs cause chromosomal aberrations as well as genomic instability, resulting in cell death. That is, a loss of gene function will result in cell sensitivity (such as the deletion of PARP1 or BRCA1/2), but the loss of function of the two genes is fatal (such as the simultaneous deletion of PARP1 and BRCA1/2). This phenomenon in which BRCAness and PARP1/2 inhibit co-killing cells is called Synthetic lethality. The theory of synergistic lethality provides new strategies and new ideas for the treatment of malignant tumors. Since then, the research of PARP1 inhibitors has entered a new era.
现有的PARP1/2抑制剂主要通过模拟PARP的底物NAD+，竞争性抑制PARP活性。大批不同骨架PARP抑制剂X-ray共晶研究为PARP1抑制剂构效关系提供了充分的支持。概括起来PARP抑制剂构效关系主要有以下几个方面：维持活性和结合力必须至少含有一个氢游离的甲酰胺结构，它可以稠合到双环结构或者“假双环”体系中，同时成为氢键受体和氢键给体；用于改变活性和理化性质的体积较大的疏水性基团等等。目前，已有多个PARP1/2抑制剂进入临床试验，其中AZD2281已被批准上市。然而，大多数PARP1/2抑制剂都存在生物利用度低，亚型选择性差的缺点。Existing PARP1/2 inhibitors competitively inhibit PARP activity primarily by mimicking the substrate NAD + of PARP. A large number of different backbone PARP inhibitor X-ray eutectic studies provide sufficient support for the structure-activity relationship of PARP1 inhibitors. In summary, the structure-activity relationship of PARP inhibitors mainly has the following aspects: maintenance activity and binding force must contain at least one hydrogen free formamide structure, which can be fused to a bicyclic structure or a "pseudobicyclic" system, and simultaneously becomes a hydrogen bond. Receptors and hydrogen bond donors; bulky hydrophobic groups for altering activity and physicochemical properties, and the like. Currently, several PARP1/2 inhibitors have entered clinical trials, of which AZD2281 has been approved for marketing. However, most PARP1/2 inhibitors have the disadvantage of low bioavailability and poor subtype selectivity.
综上，本领域尚需进一步研发PARP1/2抑制剂。In summary, there is a need in the art to further develop PARP1/2 inhibitors.
发明内容Summary of the invention
本发明的目的在于提供一种2-取代苯并咪唑-4-甲酰胺类化合物及其制备方法和应用。An object of the present invention is to provide a 2-substituted benzimidazole-4-carboxamide compound, a preparation method and application thereof.
本发明的第一方面，提供一种如通式Ia所示的化合物、R-异构体、S-异构体、或其药学上可接受的盐：According to a first aspect of the invention, there is provided a compound of the formula Ia, an R-isomer, an S-isomer, or a pharmaceutically acceptable salt thereof:
Figure PCTCN2016087368-appb-000001
Figure PCTCN2016087368-appb-000001
其中，among them,
R为氢、C1-C4的直链或支链烷基；R is hydrogen, a linear or branched alkyl group of C1-C4;
R1为氢、取代或未取代的C1-C4的直链或支链烷基、或取代或未取代的C3-C6的环烷基；R 1 is hydrogen, a substituted or unsubstituted C1-C4 linear or branched alkyl group, or a substituted or unsubstituted C3-C6 cycloalkyl group;
R2为无、卤素、取代或未取代的C1-C4的直链或支链烷基、或取代或未取代的C3-C6的环烷基；R 2 is a free, halogen, substituted or unsubstituted C1-C4 linear or branched alkyl group, or a substituted or unsubstituted C3-C6 cycloalkyl group;
Y1、Y2独立地选自取代或未取代的亚甲基、取代或未取代的亚乙基、取代或未取代的亚丙基或取代或未取代的亚丁基；Y 1 , Y 2 are independently selected from substituted or unsubstituted methylene, substituted or unsubstituted ethylene, substituted or unsubstituted propylene or substituted or unsubstituted butylene;
R3为氢、取代或未取代的C1-C4的直链或支链烷基、-C(＝O)R4、-SO2R5或取代或未取代的C3-C6的环烷基，其中，R4、R5独立地为取代或未取代的C1-C4的直链或支链烷基、取代或未取代的C3-C6的环烷基或C6-C10芳基；R 3 is hydrogen, a substituted or unsubstituted C1-C4 linear or branched alkyl group, -C(=O)R 4 , -SO 2 R 5 or a substituted or unsubstituted C3-C6 cycloalkyl group, Wherein R 4 and R 5 are independently a substituted or unsubstituted C1-C4 linear or branched alkyl group, a substituted or unsubstituted C3-C6 cycloalkyl group or a C6-C10 aryl group;
X为CR6或N，其中R6为氢、取代或未取代的C1-C4的直链或支链烷基、取代或未取代的C1-C4的直链或支链烷氧基、卤素或取代或未取代的C3-C6的环烷基；X is CR 6 or N, wherein R 6 is hydrogen, a substituted or unsubstituted C1-C4 linear or branched alkyl group, a substituted or unsubstituted C1-C4 linear or branched alkoxy group, halogen or a substituted or unsubstituted C3-C6 cycloalkyl group;
其中所述各取代独立地指具有选自下组的1-3个取代基：羟基、卤素、C1-C6直链或支链烷基、C1-C4直链或支链烷氧基、C3-C6的环烷基、C6-C10芳基、羧基；所述取代基中的C1-C6直链或支链烷基可进一步被氨基、羟基、甲氧基、乙氧基、丙氧基、异丙氧基、-CO2Me、-CO2Et、-CO2Pr或-CO2Pr-i取代。Wherein each substitution independently refers to having from 1 to 3 substituents selected from the group consisting of hydroxy, halo, C1-C6 straight or branched alkyl, C1-C4 straight or branched alkoxy, C3- a cycloalkyl group, a C6-C10 aryl group or a carboxyl group of C6; the C1-C6 linear or branched alkyl group in the substituent may be further amino, hydroxy, methoxy, ethoxy, propoxy or iso Propyl, -CO 2 Me, -CO 2 Et, -CO 2 Pr or -CO 2 Pr-i are substituted.
在具体的实施方式中，本发明提供一种通式I所示的化合物、R-异构体、S-异构体、或其药学上可接受的盐：In a specific embodiment, the invention provides a compound of formula I, an R-isomer, an S-isomer, or a pharmaceutically acceptable salt thereof:
Figure PCTCN2016087368-appb-000002
Figure PCTCN2016087368-appb-000002
其中，among them,
R1为氢、取代或未取代的C1-C4的直链或支链烷基、或取代或未取代的C3-C6的环烷基；R 1 is hydrogen, a substituted or unsubstituted C1-C4 linear or branched alkyl group, or a substituted or unsubstituted C3-C6 cycloalkyl group;
R2为无、卤素、取代或未取代的C1-C4的直链或支链烷基、或取代或未取代的C3-C6的环烷基；R 2 is a free, halogen, substituted or unsubstituted C1-C4 linear or branched alkyl group, or a substituted or unsubstituted C3-C6 cycloalkyl group;
Y1、Y2独立地选自取代或未取代的亚甲基、取代或未取代的亚乙基、取代或未取代的亚丙基或取代或未取代的亚丁基；Y 1 , Y 2 are independently selected from substituted or unsubstituted methylene, substituted or unsubstituted ethylene, substituted or unsubstituted propylene or substituted or unsubstituted butylene;
R3为氢、取代或未取代的C1-C4的直链或支链烷基、-C(＝O)R4、-SO2R5或取代或未取代的C3-C6的环烷基，其中，R4、R5独立地为取代或未取代的C1-C4的直链或支链烷基、取代或未取代的C3-C6的环烷基或C6-C10芳基；R 3 is hydrogen, a substituted or unsubstituted C1-C4 linear or branched alkyl group, -C(=O)R 4 , -SO 2 R 5 or a substituted or unsubstituted C3-C6 cycloalkyl group, Wherein R 4 and R 5 are independently a substituted or unsubstituted C1-C4 linear or branched alkyl group, a substituted or unsubstituted C3-C6 cycloalkyl group or a C6-C10 aryl group;
X为CR6或N，其中R6为氢、取代或未取代的C1-C4的直链或支链烷基或取代或未取代的C3-C6的环烷基； X is CR 6 or N, wherein R 6 is hydrogen, a substituted or unsubstituted C1-C4 linear or branched alkyl group or a substituted or unsubstituted C3-C6 cycloalkyl group;
其中所述各取代独立地指具有选自下组的1-3个取代基：羟基、卤素、C1-C6直链或支链烷基、C1-C4直链或支链烷氧基、C3-C6的环烷基、C6-C10芳基、羧基。Wherein each substitution independently refers to having from 1 to 3 substituents selected from the group consisting of hydroxy, halo, C1-C6 straight or branched alkyl, C1-C4 straight or branched alkoxy, C3- a cycloalkyl group of C6, a C6-C10 aryl group, or a carboxyl group.
在另一优选例中，R为氢、甲基、乙基。In another preferred embodiment, R is hydrogen, methyl, ethyl.
在另一优选例中，R1为氢、C1-C4的直链或支链烷基、卤素取代的C1-C4的直链或支链烷基或C3-C6的环烷基。In another preferred embodiment, R 1 is hydrogen, a C1-C4 linear or branched alkyl group, a halogen-substituted C1-C4 linear or branched alkyl group or a C3-C6 cycloalkyl group.
在另一优选例中，R1为氢、甲基、乙基、单氟甲基、三氟甲基或二氟甲基。In another preferred embodiment, R 1 is hydrogen, methyl, ethyl, monofluoromethyl, trifluoromethyl or difluoromethyl.
在另一优选例中，R2为氢、氟、氯、溴、C1-C4的直链或支链烷基、或C3-C6的环烷基。In another preferred embodiment, R 2 is hydrogen, fluorine, chlorine, bromine, a C1-C4 linear or branched alkyl group, or a C3-C6 cycloalkyl group.
在另一优选例中，R2为无、氟、氯、甲基、乙基。In another preferred embodiment, R 2 is none, fluorine, chlorine, methyl, ethyl.
在另一优选例中，R3为氢、C1-C4的直链或支链烷基、-C(＝O)R4或-SO2R5，其中，R4、R5独立地为取代或未取代的C1-C4的直链或支链烷基、C3-C6的环烷基或苯基，所述取代是指具有选自下组的1-2个取代基：羟基、卤素、C1-C4直链或支链烷氧基。In another preferred embodiment, R 3 is hydrogen, a C1-C4 linear or branched alkyl group, -C(=O)R 4 or -SO 2 R 5 , wherein R 4 and R 5 are independently substituted. Or an unsubstituted C1-C4 linear or branched alkyl group, a C3-C6 cycloalkyl group or a phenyl group, said substituent having 1-2 substituents selected from the group consisting of hydroxy, halogen, C1 -C4 linear or branched alkoxy group.
在另一优选例中，R3为氢、甲基、乙基、乙酰基、环丙甲酰基、苯甲酰基、甲磺酰基、-COCH2OCH2CH2CH3。In another preferred embodiment, R 3 is hydrogen, methyl, ethyl, acetyl, cyclopropanoyl, benzoyl, methylsulfonyl, -COCH 2 OCH 2 CH 2 CH 3 .
在另一优选例中，X为CR6或N，其中R6为氢、C1-C4的直链或支链烷基或C3-C6的环烷基。In another preferred embodiment, X is CR 6 or N, wherein R 6 is hydrogen, a C1-C4 linear or branched alkyl group or a C3-C6 cycloalkyl group.
在另一优选例中，X为CH、C(CH3)、C(CH2CH3)或N。In another preferred embodiment, X is CH, C(CH 3 ), C(CH 2 CH 3 ) or N.
在另一优选例中，Y1为取代或未取代的亚甲基、Y2取代或未取代的亚乙基；或者Y1为取代或未取代的亚乙基、Y2为取代或未取代的亚甲基，其中In another preferred embodiment, Y 1 is a substituted or unsubstituted methylene group, a Y 2 -substituted or unsubstituted ethylene group; or Y 1 is a substituted or unsubstituted ethylene group, and Y 2 is a substituted or unsubstituted group. Methylene, of which
所述各取代独立地指具有选自下组的1-2个取代基：羟基、卤素、C1-C4直链或支链烷基、C1-C4直链或支链烷氧基、C3-C6的环烷基；其中的C1-C4直链或支链烷基可进一步被氨基、羟基、甲氧基、乙氧基、丙氧基、异丙氧基、-CO2Me、-CO2Et、-CO2Pr或-CO2Pr-i取代。Each of the substituents independently means having 1-2 substituents selected from the group consisting of hydroxy, halogen, C1-C4 straight or branched alkyl, C1-C4 straight or branched alkoxy, C3-C6 a cycloalkyl group; wherein the C1-C4 straight or branched alkyl group may be further amino, hydroxy, methoxy, ethoxy, propoxy, isopropoxy, -CO 2 Me, -CO 2 Et , -CO 2 Pr or -CO 2 Pr-i substituted.
在另一优选例中，Y1为取代或未取代的亚甲基、Y2取代或未取代的亚乙基；或者Y1为取代或未取代的亚乙基、Y2为取代或未取代的亚甲基，其中，所述取代基独立地指具有选自下组的1-2个取代基：甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、环丙基、环丁基、-C(CH3)2NH2、-C(CH3)2OH、-C(CH3)2CO2CH3、-C(CH3)2OC2H5或-C(CH3)2OCH3。在另一优选例中，所述化合物为实施例制备的I-1到I-33，I-36到I-45中的任一化合物。In another preferred embodiment, Y 1 is a substituted or unsubstituted methylene group, a Y 2 -substituted or unsubstituted ethylene group; or Y 1 is a substituted or unsubstituted ethylene group, and Y 2 is a substituted or unsubstituted group. a methylene group, wherein the substituent independently means having 1 to 2 substituents selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tertiary. Butyl, cyclopropyl, cyclobutyl, -C(CH 3 ) 2 NH 2 , -C(CH 3 ) 2 OH, -C(CH 3 ) 2 CO 2 CH 3 , -C(CH 3 ) 2 OC 2 H 5 or -C(CH 3 ) 2 OCH 3 . In another preferred embodiment, the compound is any one of the compounds I-1 to I-33, I-36 to I-45 prepared in the examples.
本发明的第二方面，提供第一方面所述的化合物的制备方法，包括通式IIa所述的化合物发生氨解得到通式Ia所述的化合物的步骤：According to a second aspect of the present invention, there is provided a process for the preparation of a compound according to the first aspect, which comprises the step of obtaining a compound of the formula Ia by aminolysis of a compound of the formula IIa:
Figure PCTCN2016087368-appb-000003
Figure PCTCN2016087368-appb-000003
Figure PCTCN2016087368-appb-000004
Figure PCTCN2016087368-appb-000004
其中，R、R1、R2、Y1、Y2、R3、X的定义如第一方面所述，R7选自C1-C4的直链或支链烷基。Wherein R, R 1 , R 2 , Y 1 , Y 2 , R 3 , X are as defined in the first aspect, and R 7 is selected from a linear or branched alkyl group of C1-C4.
在具体的实施方式中，上述方法包括包括通式II所述的化合物发生氨解得到通式I所述的化合物的步骤：In a specific embodiment, the above method comprises the steps of: subjecting the compound of Formula II to aminolysis to give a compound of Formula I:
Figure PCTCN2016087368-appb-000005
Figure PCTCN2016087368-appb-000005
其中，R1、R2、Y1、Y2、R3、X的定义如第一方面所述。Wherein R 1 , R 2 , Y 1 , Y 2 , R 3 , X are as defined in the first aspect.
本发明的第三方面，提供一种药物组合物，包括第一方面所述的化合物、R-异构体、S-异构体、或其药学上可接受的盐；以及A third aspect of the invention provides a pharmaceutical composition comprising the compound of the first aspect, the R-isomer, the S-isomer, or a pharmaceutically acceptable salt thereof;
药学上可接受的载体。A pharmaceutically acceptable carrier.
本发明的第四方面，提供第一方面所述的化合物或第三方面所述的药物组合物的用途，用于制备：A fourth aspect of the invention provides the use of the compound of the first aspect or the pharmaceutical composition of the third aspect for the preparation of:
(1)PARP1抑制剂；(1) a PARP1 inhibitor;
(2)PARP2抑制剂；(2) PARP2 inhibitors;
(3)预防和/或治疗肿瘤的药物；(3) drugs for preventing and/or treating tumors;
(4)抗炎药物；和/或(4) anti-inflammatory drugs; and/or
(5)预防和/或治疗与PARP相关疾病的药物。(5) A drug for preventing and/or treating a disease associated with PARP.
本发明的第五方面，提供一种第一方面所述的化合物的中间体，结构如通式IIa所示：According to a fifth aspect of the invention, there is provided an intermediate of the compound of the first aspect, which has the structure shown in the formula IIa:
Figure PCTCN2016087368-appb-000006
Figure PCTCN2016087368-appb-000006
其中，among them,
R1、R2、Y1、Y2、R3、X的定义如第一方面所述，R7选自C1-C4的直链或支链烷基，优选自甲基、乙基和叔丁基。R 1 , R 2 , Y 1 , Y 2 , R 3 , X are as defined in the first aspect, and R 7 is selected from C1-C4 straight or branched alkyl groups, preferably from methyl, ethyl and untertiary Butyl.
在具体的实施方式中，式IIa所示化合物如通式II所示： In a specific embodiment, the compound of Formula IIa is as shown in Formula II:
Figure PCTCN2016087368-appb-000007
Figure PCTCN2016087368-appb-000007
其中，among them,
R1、R2、Y1、Y2、R3、X的定义如第一方面所述。The definitions of R 1 , R 2 , Y 1 , Y 2 , R 3 , and X are as described in the first aspect.
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。It is to be understood that within the scope of the present invention, the various technical features of the present invention and the various technical features specifically described hereinafter (as in the embodiments) may be combined with each other to constitute a new or preferred technical solution. Due to space limitations, we will not repeat them here.
具体实施方式detailed description
本申请的发明人经过广泛而深入地研究，首次设计并合成了一类含硫原子的苯并咪唑-4-甲酰胺类PARP1/2抑制剂。本发明所涉及化合物具有明确的构效关系，且显示了很好的体内外抗肿瘤活性。该类新型PARP1/2抑制剂有望成为新型抗肿瘤药物。在此基础上完成了本发明。The inventors of the present application have extensively and intensively studied to design and synthesize a class of benzimidazole-4-carboxamide PARP1/2 inhibitors containing sulfur atoms for the first time. The compounds of the present invention have a well-defined structure-activity relationship and exhibit excellent anti-tumor activity in vitro and in vivo. This new type of PARP1/2 inhibitor is expected to become a new anti-tumor drug. The present invention has been completed on this basis.
术语the term
本发明的上下文中，术语“烷基”表示饱和的线性或支链烃部分，例如-CH3或-CH(CH3)2。术语“烷氧基”表示-O-(烷基)基团。术语“环烷基”表示饱和的环状烃基部分，例如环丙基、环己基。术语“芳基”表示包含一个或多个芳环的烃基部分。芳基部分的例子包括苯基(Ph)、萘基等。In the context of this invention, the term "alkyl" means a saturated linear or branched hydrocarbon moiety, such as -CH 3 or -CH (CH 3) 2. The term "alkoxy" denotes an -O-(alkyl) group. The term "cycloalkyl" denotes a saturated cyclic hydrocarbyl moiety such as cyclopropyl, cyclohexyl. The term "aryl" refers to a hydrocarbyl moiety containing one or more aromatic rings. Examples of the aryl moiety include phenyl (Ph), naphthyl and the like.
除非另外说明，本文所述的烷基、烷氧基、环烷基和芳基同时包括取代的和未取代的部分。烷基、烷氧基、环烷基和芳基上可能的取代基包括，但不限于：C1-C6烷基、C2-C6烯基、C2-C6炔基、C3-C10环烷基、C1-C20杂环烷基、C1-C10烷氧基、C6-C10芳基、氨基、C1-C10烷基氨基、C1-C20二烷基氨基、氰基、硝基和羧基。Unless otherwise stated, the alkyl, alkoxy, cycloalkyl and aryl groups described herein include both substituted and unsubstituted moieties. Possible substituents on alkyl, alkoxy, cycloalkyl and aryl groups include, but are not limited to, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C10 cycloalkyl, C1 -C20 heterocycloalkyl, C1-C10 alkoxy, C6-C10 aryl, amino, C1-C10 alkylamino, C1-C20 dialkylamino, cyano, nitro and carboxy.
通式Ia所示的化合物a compound of formula Ia
本发明提供的通式Ia所示的化合物，结构如下：The compound of the formula Ia provided by the present invention has the following structure:
Figure PCTCN2016087368-appb-000008
Figure PCTCN2016087368-appb-000008
R、R1、R2、Y1、Y2、R3、X的定义如前所示。The definitions of R, R 1 , R 2 , Y 1 , Y 2 , R 3 and X are as defined above.
本发明的化合物具有不对称中心、手性轴和手性平面，并且可以以外消旋体、R- 异构体或S-异构体的形式存在。本领域技术人员能够采用常规技术手段由外消旋体拆分获得R-异构体和/或S-异构体。The compounds of the invention have asymmetric centers, chiral axes and chiral planes, and can be racemic, R- The form of the isomer or S-isomer is present. Those skilled in the art will be able to resolve the R-isomer and/or the S-isomer from the racemate by conventional techniques.
本发明提供还通式Ia化合物的药学上可而接受的盐，具体地为通式Ia化合物与无机酸或有机酸反应形成常规的药学上可接受的盐。例如，常规的药学上可接受的盐可通过通式Ia化合物与无机酸或有机酸反应制得，所述无机酸包括盐酸、氢溴酸、硫酸、硝酸、胺基磺酸和磷酸等，以及所述有机酸包括柠檬酸、酒石酸、乳酸、丙酮酸、乙酸、苯磺酸、对甲苯磺酸、甲磺酸、萘磺酸、乙磺酸、萘二磺酸、马来酸、苹果酸、丙二酸、富马酸、琥珀酸、丙酸、草酸、三氟乙酸、硬酯酸、扑酸、羟基马来酸、苯乙酸、苯甲酸、水杨酸、谷氨酸、抗坏血酸、对胺基苯磺酸、2-乙酰氧基苯甲酸和羟乙磺酸等；或者通式Ia化合物与无机碱形成的钠盐、钾盐、钙盐、铝盐或铵盐；或者通式Ia化合物与有机碱形成的甲胺盐、乙胺盐或乙醇胺盐。The invention provides a pharmaceutically acceptable salt of a compound of formula Ia, in particular a compound of formula Ia, which is reacted with an inorganic or organic acid to form a conventional pharmaceutically acceptable salt. For example, a conventional pharmaceutically acceptable salt can be prepared by reacting a compound of the formula Ia with an inorganic or organic acid, including hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, aminosulfonic acid, phosphoric acid, and the like, and The organic acid includes citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, naphthalenesulfonic acid, ethanesulfonic acid, naphthalene disulfonic acid, maleic acid, malic acid, Malonic acid, fumaric acid, succinic acid, propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pamoic acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, salicylic acid, glutamic acid, ascorbic acid, p-amine a base, a potassium salt, a calcium salt, an aluminum salt or an ammonium salt formed from a compound of the formula Ia with an inorganic base; or a compound of the formula Ia with a benzsulfonic acid, 2-acetoxybenzoic acid and isethionate; A methylamine salt, ethylamine salt or ethanolamine salt formed by an organic base.
在具体的实施方式中，本发明提供通式I所示的化合物、R-异构体、S-异构体、或其药学上可接受的盐：In a specific embodiment, the invention provides a compound of formula I, an R-isomer, an S-isomer, or a pharmaceutically acceptable salt thereof:
Figure PCTCN2016087368-appb-000009
Figure PCTCN2016087368-appb-000009
R1、R2、Y1、Y2、R3、X的定义如前所示。The definitions of R 1 , R 2 , Y 1 , Y 2 , R 3 and X are as indicated above.
制备方法Preparation
(1)当R1为氢，R3为氢时，本发明式I-1，I-4，I-6～I-8，I-15，I-17～I-29和I-36～I-40表示的化合物的合成方案如下。(1) When R 1 is hydrogen and R 3 is hydrogen, the formulae I-1, I-4, I-6 to I-8, I-15, I-17 to I-29 and I-36 of the present invention The synthesis scheme of the compound represented by I-40 is as follows.
Figure PCTCN2016087368-appb-000010
Figure PCTCN2016087368-appb-000010
邻苯二胺甲酸酯类化合物1和醛类衍生物2在亚硫酸氢钠或者其他路易斯酸的存在下发生缩合环化并氧化生成化合物3；化合物3在三氟醋酸等酸的存在下经脱Boc保护基生成化合物4，化合物4通过氨解试剂氨解生成化合物I-1，I-4，I-6～I-8，I-15，I-17～I-29，I-36～I-40。The o-phenylenedicarboxylate compound 1 and the aldehyde derivative 2 are condensed and cyclized in the presence of sodium hydrogen sulfite or other Lewis acid and oxidized to form the compound 3; the compound 3 is removed in the presence of an acid such as trifluoroacetic acid. The Boc protecting group generates the compound 4, and the compound 4 is aminolyzed by the aminolysis reagent to give the compound I-1, I-4, I-6 to I-8, I-15, I-17 to I-29, I-36 to I. -40.
(2)当R1为氢，R3为氢时，本发明式I-1，I-4，I-6～I-8，I-15，I-17～I-29和I-36～I-40 表示的化合物也可通过如下合成方案制备。(2) When R 1 is hydrogen and R 3 is hydrogen, the formulas I-1, I-4, I-6 to I-8, I-15, I-17 to I-29 and I-36 of the present invention The compound represented by I-40 can also be produced by the following synthesis scheme.
Figure PCTCN2016087368-appb-000011
Figure PCTCN2016087368-appb-000011
化合物3先通过氨解试剂氨解生成化合物5，化合物5在盐酸或者三氟醋酸等酸的作用下经脱Boc保护基制得I-1，I-4，I-6～I-8，I-15，I-17～I-29，I-36～I-40。Compound 3 is firstly aminolyzed by an aminolysis reagent to form compound 5, and compound 5 is obtained by removing the Boc protecting group under the action of an acid such as hydrochloric acid or trifluoroacetic acid to obtain I-1, I-4, I-6 to I-8, I. -15, I-17 to I-29, I-36 to I-40.
(3)当R1为氢，R3为氢时，本发明式I-1，I-4，I-6～I-8，I-15，I-17～I-29和I-36～I-40表示的化合物也可通过如下合成方案制备。(3) When R 1 is hydrogen and R 3 is hydrogen, the formulas I-1, I-4, I-6 to I-8, I-15, I-17 to I-29 and I-36 of the present invention The compound represented by I-40 can also be produced by the following synthetic scheme.
Figure PCTCN2016087368-appb-000012
Figure PCTCN2016087368-appb-000012
邻苯二胺甲酸酯类化合物1与氨解试剂反应得到化合物6，化合物6与化合物2在亚硫酸氢钠或者其他路易斯酸的存在下发生缩合环化并氧化生成化合物5，化合物5通过三氟乙酸或盐酸等酸的作用下脱Boc保护基生成化合物I-1，I-4，I-6～I-8，I-15，I-17～I-29，I-36～I-40。The o-phenylenediaminecarboxylate compound 1 is reacted with an aminolysis reagent to obtain a compound 6, which is condensed and cyclized in the presence of sodium hydrogen sulfite or other Lewis acid to form a compound 5, and the compound 5 is passed through a trifluorocarbon. The Boc protecting group is deactivated by the action of an acid such as acetic acid or hydrochloric acid to give the compound I-1, I-4, I-6 to I-8, I-15, I-17 to I-29, and I-36 to I-40.
(4)当R1不为氢，R3为氢时，化合物I-30～I-33和I-41～43的制备方法如下：(4) When R 1 is not hydrogen and R 3 is hydrogen, the preparation methods of the compounds I-30 to I-33 and I-41 to 43 are as follows:
Figure PCTCN2016087368-appb-000013
Figure PCTCN2016087368-appb-000013
化合物3与R1L或二氟氯乙酸钠在碱作用下生成化合物7，化合物7在三氟乙酸或盐酸等酸的作用下经脱Boc保护基生成化合物8，化合物8在氨解试剂作用下生成化合物I-30～I-33和I-41～43，其中所述L为卤素。Compound 3 and R 1 L or sodium difluorochloroacetate form compound 7 under the action of a base, and compound 7 is decarboxylated to form compound 8 under the action of an acid such as trifluoroacetic acid or hydrochloric acid, and compound 8 is under the action of an aminolysis reagent. Compounds I-30 to I-33 and I-41 to 43 are produced, wherein the L is a halogen.
(5)当R1不为氢，R3为氢时，化合物I-30～I-33和I-41～43的制备也可以通过如下方法实施：(5) When R 1 is not hydrogen and R 3 is hydrogen, the preparation of the compounds I-30 to I-33 and I-41 to 43 can also be carried out by the following method:
Figure PCTCN2016087368-appb-000014
Figure PCTCN2016087368-appb-000014
化合物5与R1L或二氟氯乙酸钠在碱作用下生成化合物9，化合物9在三氟乙酸或盐酸等酸的作用下经脱Boc保护基生成化合物I-30～I-33和I-41～43，其中所述L为卤素。Compound 5 and R 1 L or sodium difluorochloroacetate form compound 9 under the action of a base, and compound 9 is decarboxylated to form compounds I-30 to I-33 and I- under the action of an acid such as trifluoroacetic acid or hydrochloric acid. 41 to 43, wherein L is a halogen.
(6)当R1为氢，R3不为氢时，化合物I-2，I-3，I-5，I-9～I-14，I-16表示的化合物的合成方案如下。(6) When R 1 is hydrogen and R 3 is not hydrogen, the synthesis scheme of the compound represented by the compound I-2, I-3, I-5, I-9 to I-14, and I-16 is as follows.
Figure PCTCN2016087368-appb-000015
Figure PCTCN2016087368-appb-000015
化合物4与R3L反应得到化合物9，再氨解生成化合物I-2，I-3，I-5，I-9～I-14，I-16。Compound 4 is reacted with R 3 L to give compound 9, which is then aminolyzed to give compound I-2, I-3, I-5, I-9 to I-14, I-16.
上述(1)～(6)反应中，R、R1﹑R2﹑R3﹑X﹑Y1和Y2如前定义所述；所述氨基试剂选自氨的甲醇溶液﹑氨的乙醇溶液﹑氨的水溶液﹑甲胺水溶液﹑甲胺醇溶液﹑乙胺水溶液或乙胺醇溶液。In the above reactions (1) to (6), R, R 1 , R 2 , R 3 , X, Y 1 and Y 2 are as defined above; the amino reagent is selected from the group consisting of ammonia methanol solution and ammonia ethanol solution. An aqueous solution of ammonia, an aqueous solution of methylamine, a solution of methylamine, an aqueous solution of ethylamine or a solution of ethylamine.
其中中间体1可由商业获得或如下步骤制备：Wherein intermediate 1 is commercially available or is prepared as follows:
Figure PCTCN2016087368-appb-000016
Figure PCTCN2016087368-appb-000016
靛红类化合物11硝化生成硝基化合物，硝基化合物在过氧化物如双氧水的存在下发生Baeyer-Villiger氧化并碱性条件下水解开环生成羧酸化合物，在酸的催化下，羧酸化合物酯化生成酯，经钯碳氢化或其他还原剂如铁粉，锌粉等存在下还原生成化合物1；The ruthenium compound 11 is nitrated to form a nitro compound, and the nitro compound is oxidized by Baeyer-Villiger in the presence of a peroxide such as hydrogen peroxide and hydrolyzed and opened under basic conditions to form a carboxylic acid compound, and the carboxylic acid compound is esterified under acid catalysis. Forming an ester, which is reduced by palladium hydrocarbonation or other reducing agent such as iron powder, zinc powder or the like to form compound 1;
醛类中间体2可由商业获得或者通过如下步骤制备：The aldehyde intermediate 2 can be obtained commercially or by the following procedure:
Figure PCTCN2016087368-appb-000017
Figure PCTCN2016087368-appb-000017
化合物13与NBS反应得到化合物12，化合物12在正丁基锂或叔丁基锂/无水DMF/无水THF的条件下得到化合物2。Compound 13 is reacted with NBS to give compound 12, which is obtained under the conditions of n-butyllithium or tert-butyllithium/anhydrous DMF/anhydrous THF.
除非另行定义，文中所使用的所有专业与科学用语与本领域熟练人员所熟悉的意义相同。此外，任何与所记载内容相似或均等的方法及材料皆可应用于本发明方法中。文中所述的较佳实施方法与材料仅作示范之用。 Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those skilled in the art. In addition, any methods and materials similar or equivalent to those described may be employed in the methods of the invention. The preferred embodiments and materials described herein are for illustrative purposes only.
药物组合物Pharmaceutical composition
本发明还提供了一种药物组合物，它包含安全有效量范围内的活性成分，以及药学上可接受的载体。The invention also provides a pharmaceutical composition comprising an active ingredient in a safe and effective amount, together with a pharmaceutically acceptable carrier.
本发明所述的“活性成分”是指本发明所述的通式I所示的2-取代苯并咪唑-4-甲酰胺类化合物。The "active ingredient" as used in the present invention means a 2-substituted benzimidazole-4-carboxamide compound represented by the formula I according to the present invention.
本发明的2-取代苯并咪唑-4-甲酰胺类化合物和药物组合物，可以用作PARP1抑制剂或PARP2抑制剂；可以用于制备预防和/或治疗与聚腺苷二磷酸核糖聚合酶PARP相关疾病的药物；可以用于制备预防和/或治疗肿瘤的药物；还可以用于制备抗炎药物。The 2-substituted benzimidazole-4-carboxamides and pharmaceutical compositions of the present invention can be used as PARP1 inhibitors or PARP2 inhibitors; can be used for the preparation of prophylactic and/or therapeutic agents with poly ADP-ribose polymerase A drug for a PARP-related disease; a drug which can be used for the preparation of a prophylactic and/or therapeutic tumor; and an anti-inflammatory drug.
在另一优选例中，所述与聚腺苷二磷酸核糖聚合酶PARP相关疾病为肿瘤、炎症以及局部缺血-再灌注并发的疾病如心血管疾病，糖尿病，风湿性关节炎，内毒素性休克，中风等。In another preferred embodiment, the disease associated with poly ADP-ribose polymerase PARP is tumor, inflammation, and ischemic-reperfusion complicated diseases such as cardiovascular disease, diabetes, rheumatoid arthritis, endotoxin Shock, stroke, etc.
在另一优选例中，所述肿瘤为卵巢癌、乳腺癌、胃癌、前列腺癌、肺癌、晚期实体瘤、神经胶质瘤、黑色素瘤、结肠癌、胰腺癌、复发血液癌、尤文氏肉瘤、胰腺癌、子宫内膜癌等。In another preferred embodiment, the tumor is ovarian cancer, breast cancer, gastric cancer, prostate cancer, lung cancer, advanced solid tumor, glioma, melanoma, colon cancer, pancreatic cancer, recurrent hematological cancer, Ewing's sarcoma, Pancreatic cancer, endometrial cancer, etc.
在另一优选例中，所述肿瘤为同源重组修复缺陷的肿瘤，即BRCA1或BRCA2缺失或突变的肿瘤，如卵巢癌、乳腺癌、前列腺癌、胃癌、胰腺癌、宫颈癌、神经胶质瘤、尤文氏肉瘤等。In another preferred embodiment, the tumor is a tumor defective in homologous recombination repair, that is, a tumor with BRCA1 or BRCA2 deletion or mutation, such as ovarian cancer, breast cancer, prostate cancer, gastric cancer, pancreatic cancer, cervical cancer, glial Tumor, Ewing's sarcoma, etc.
“安全有效量”指的是：活性成分的量足以明显改善病情，而不至于产生严重的副作用。通常，药物组合物含有1-2000mg活性成分/剂，更佳地，含有10-200mg活性成分/剂。较佳地，所述的“一剂”为一个药片。By "safe and effective amount" is meant that the amount of active ingredient is sufficient to significantly improve the condition without causing serious side effects. In general, the pharmaceutical compositions contain from 1 to 2000 mg of active ingredient per dose, more preferably from 10 to 200 mg of active ingredient per dose. Preferably, the "one dose" is a tablet.
“药学上可接受的载体”指的是：一种或多种相容性固体或液体填料或凝胶物质，它们适合于人使用，而且必须有足够的纯度和足够低的毒性。“相容性”在此指的是组合物中各组份能和本发明的活性成分以及它们之间相互掺和，而不明显降低活性成分的药效。药学上可以接受的载体部分例子有纤维素及其衍生物(如羧甲基纤维素钠、乙基纤维素钠、纤维素乙酸酯等)、明胶、滑石、固体润滑剂(如硬脂酸、硬脂酸镁)、硫酸钙、植物油(如豆油、芝麻油、花生油、橄榄油等)、多元醇(如丙二醇、甘油、甘露醇、山梨醇等)、乳化剂
Figure PCTCN2016087368-appb-000018
润湿剂(如十二烷基硫酸钠)、着色剂、调味剂、稳定剂、抗氧化剂、防腐剂、无热原水等。"Pharmaceutically acceptable carrier" means: one or more compatible solid or liquid fillers or gel materials which are suitable for human use and which must be of sufficient purity and of sufficiently low toxicity. By "compatibility" it is meant herein that the components of the composition are capable of intermingling with the active ingredients of the present invention and with respect to each other without significantly reducing the efficacy of the active ingredients. Examples of pharmaceutically acceptable carriers are cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid). , magnesium stearate), calcium sulfate, vegetable oil (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyol (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifier
Figure PCTCN2016087368-appb-000018
Wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.
在另一优选例中，本发明通式I所示的2-取代苯并咪唑-4-甲酰胺类化合物可与大分子化合物或高分子通过非键合作用形成复合物。在另一优选例中，本发明通式I所示的2-取代苯并咪唑-4-甲酰胺类化合物作为小分子还可通过化学键与大分子化合物或高分子相连接。所述大分子化合物可以是生物大分子如高聚糖、蛋白、核酸、多肽等。In another preferred embodiment, the 2-substituted benzimidazole-4-carboxamide compound of the formula I of the present invention can form a complex with a macromolecular compound or a polymer by non-bonding. In another preferred embodiment, the 2-substituted benzimidazole-4-carboxamide compound of the formula I of the present invention can be linked as a small molecule to a macromolecular compound or a polymer by a chemical bond. The macromolecular compound may be a biological macromolecule such as a polysaccharide, a protein, a nucleic acid, a polypeptide, or the like.
本发明的活性成分或药物组合物的施用方式没有特别限制，代表性的施用方式包括(但并不限于)：口服、瘤内、直肠、肠胃外(静脉内、肌肉内或皮下)等。The administration form of the active ingredient or the pharmaceutical composition of the present invention is not particularly limited, and representative administration forms include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous) and the like.
用于口服给药的固体剂型包括胶囊剂、片剂、丸剂、散剂和颗粒剂。Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.
在这些固体剂型中，活性成分与至少一种常规惰性赋形剂(或载体)混合，如柠檬酸钠或磷酸二钙，或与下述成分混合：(a)填料或增容剂，例如，淀粉、乳糖、蔗糖、葡萄糖、甘 露醇和硅酸；(b)粘合剂，例如，羟甲基纤维素、藻酸盐、明胶、聚乙烯基吡咯烷酮、蔗糖和阿拉伯胶；(c)保湿剂，例如，甘油；(d)崩解剂，例如，琼脂、碳酸钙、马铃薯淀粉或木薯淀粉、藻酸、某些复合硅酸盐、和碳酸钠；(e)缓溶剂，例如石蜡；(f)吸收加速剂，例如，季胺化合物；(g)润湿剂，例如鲸蜡醇和单硬脂酸甘油酯；(h)吸附剂，例如，高岭土；和(i)润滑剂，例如，滑石、硬脂酸钙、硬脂酸镁、固体聚乙二醇、十二烷基硫酸钠，或其混合物。胶囊剂、片剂和丸剂中，剂型也可包含缓冲剂。In these solid dosage forms, the active ingredient is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or mixed with: (a) a filler or compatibilizer, for example, Starch, lactose, sucrose, glucose, sweet (l) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerin; (d) collapse Decomposing agents, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) a slow solvent such as paraffin; (f) an absorption accelerator, for example, a quaternary amine a compound; (g) a wetting agent such as cetyl alcohol and glyceryl monostearate; (h) an adsorbent such as kaolin; and (i) a lubricant such as talc, calcium stearate, magnesium stearate , solid polyethylene glycol, sodium lauryl sulfate, or a mixture thereof. In capsules, tablets and pills, the dosage form may also contain a buffer.
所述的固体剂型还可采用包衣和壳材制备，如肠衣和其它本领域公知的材料。它们可包含不透明剂，并且，这种组合物中活性成分的释放可以延迟的方式在消化道内的某一部分中释放。可采用的包埋组分的实例是聚合物质和蜡类物质。The solid dosage forms can also be prepared with coatings and shell materials, such as casings and other materials known in the art. They may contain opacifying agents and the release of the active ingredient in such compositions may be released in a portion of the digestive tract in a delayed manner. Examples of embedding components that can be employed are polymeric and waxy materials.
用于口服给药的液体剂型包括药学上可接受的乳液、溶液、悬浮液、糖浆或酊剂。除了活性成分外，液体剂型可包含本领域中常规采用的惰性稀释剂，如水或其它溶剂，增溶剂和乳化剂，例知，乙醇、异丙醇、碳酸乙酯、乙酸乙酯、丙二醇、1,3-丁二醇、二甲基甲酰胺以及油，特别是棉籽油、花生油、玉米胚油、橄榄油、蓖麻油和芝麻油或这些物质的混合物等。除了这些惰性稀释剂外，组合物也可包含助剂，如润湿剂、乳化剂和悬浮剂、甜味剂、矫味剂和香料。Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or elixirs. In addition to the active ingredient, the liquid dosage form may contain inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or a mixture of these substances. In addition to these inert diluents, the compositions may contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and perfumes.
除了活性成分外，悬浮液可包含悬浮剂，例如，乙氧基化异十八烷醇、聚氧乙烯山梨醇和脱水山梨醇酯、微晶纤维素、甲醇铝和琼脂或这些物质的混合物等。In addition to the active ingredient, the suspension may contain suspending agents, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these and the like.
用于肠胃外注射的组合物可包含生理上可接受的无菌含水或无水溶液、分散液、悬浮液或乳液，和用于重新溶解成无菌的可注射溶液或分散液的无菌粉末。适宜的含水和非水载体、稀释剂、溶剂或赋形剂包括水、乙醇、多元醇及其适宜的混合物。Compositions for parenteral injection may comprise a physiologically acceptable sterile aqueous or nonaqueous solution, dispersion, suspension or emulsion, and a sterile powder for reconstitution into a sterile injectable solution or dispersion. Suitable aqueous and nonaqueous vehicles, diluents, solvents or vehicles include water, ethanol, polyols, and suitable mixtures thereof.
本发明化合物可以单独给药，或者与其他治疗药物(如化疗药)联合给药。The compounds of the invention may be administered alone or in combination with other therapeutic agents, such as chemotherapeutic agents.
使用药物组合物时，是将安全有效量的本发明化合物适用于需要治疗的哺乳动物(如人)，其中施用时剂量为药学上认为的有效给药剂量，对于60kg体重的人而言，日给药剂量通常为1～2000mg，优选20～500mg。当然，具体剂量还应考虑给药途径、病人健康状况等因素，这些都是熟练医师技能范围之内的。When a pharmaceutical composition is used, a safe and effective amount of a compound of the invention is administered to a mammal (e.g., a human) in need of treatment wherein the dosage is a pharmaceutically effective effective dosage, for a 60 kg body weight, The dose to be administered is usually from 1 to 2000 mg, preferably from 20 to 500 mg. Of course, specific doses should also consider factors such as the route of administration, the health of the patient, etc., which are within the skill of the skilled physician.
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件如Sambrook等人，分子克隆：实验室手册(New York:Cold Spring Harbor Laboratory Press,1989)中所述的条件，或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数按重量计算。The invention is further illustrated below in conjunction with specific embodiments. It is to be understood that the examples are not intended to limit the scope of the invention. The experimental methods in the following examples which do not specify the specific conditions are usually carried out according to the conditions described in conventional conditions such as Sambrook et al., Molecular Cloning: Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the manufacturer. The suggested conditions. Percentages and parts are by weight unless otherwise stated.
所有实施例中，1H NMR由Varian Mercury-300或Varian Mercury-400型核磁共振仪记录，13C NMR由Varian Mercury-400或Varian Mercury-500型或Varian Mercury-600型核磁共振仪记录，化学位移以δ(ppm)表示；质谱由Finnigan/MAT-95(EI)与Finnigan LCQ/DECA and Micromass Ultra Q-TOF(ESI)型质谱仪记录；分离用硅胶为200-300目。 In all examples, 1 H NMR was recorded by a Varian Mercury-300 or Varian Mercury-400 nuclear magnetic resonance apparatus, and 13 C NMR was recorded by a Varian Mercury-400 or Varian Mercury-500 or Varian Mercury-600 nuclear magnetic resonance apparatus, chemistry. The displacement is expressed in δ (ppm); the mass spectrum is recorded by Finnigan/MAT-95 (EI) and Finnigan LCQ/DECA and Micromass Ultra Q-TOF (ESI) mass spectrometer; the separation silica gel is 200-300 mesh.
实施例1. 2-(4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-1的制备：Example 1. 2-(4,5,6,7-Tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide, I-1 Preparation:
步骤1. 6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯：Step 1. 6,7-Dihydrothieno[3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000019
Figure PCTCN2016087368-appb-000019
称取4,5,6,7-四氢噻吩[3,2-c]吡啶盐酸盐(10g，56.9mmol)于反应瓶(500mL)中，加二氯甲烷(300mL)。冰浴下加三乙胺(8.7mL)，缓慢地加二碳酸二叔丁酯(13.7g，62.8mmol)的二氯甲烷(20mL)溶液。滴加完毕后，1小时跟踪反应，原料反应完全。将反应液依次用水洗、饱和NaCl水溶液洗涤，保留有机层，用无水Na2SO4干燥，过滤减压浓缩，粗产品经柱层析(石油醚：乙酸乙酯＝20：1)得无色透明油状物，后固化得13.5g白色固体，产率为99％。4,5,6,7-Tetrahydrothiophene [3,2-c]pyridine hydrochloride (10 g, 56.9 mmol) was weighed in a reaction flask (500 mL) and dichloromethane (300 mL). Triethylamine (8.7 mL) was added under ice-cooling, and a solution of di-tert-butyl dicarbonate (13.7 g, 62.8 mmol) in dichloromethane (20 mL). After the completion of the dropwise addition, the reaction was followed for 1 hour, and the reaction of the starting material was completed. The reaction mixture was successively washed with water saturated aqueous NaCl solution, the organic layer was retained, dried over anhydrous Na 2 SO 4, filtered and concentrated under reduced pressure and the crude product purified by column chromatography (petroleum ether: ethyl acetate = 20: 1) to give None A clear oily solid which was post-solidified to give 13.5 g of a white solid.
1H NMR(300MHz,Chloroform-d)δ7.12(d,J＝5.1Hz,1H),6.78(d,J＝5.1Hz,1H),4.50(s,2H),3.71(t,J＝5.3Hz,2H),2.84(t,J＝5.3Hz,2H),1.48(s,9H)；MS(ESI):m/z 240[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ7.12 (d, J = 5.1Hz, 1H), 6.78 (d, J = 5.1Hz, 1H), 4.50 (s, 2H), 3.71 (t, J = 5.3 Hz, 2H), 2.84 (t, J = 5.3 Hz, 2H), 1.48 (s, 9H); MS (ESI): m/z 240 [M+H] + .
步骤2. 2-溴-6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯：Step 2. 2-Bromo-6,7-dihydrothieno[3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000020
Figure PCTCN2016087368-appb-000020
向6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯(2.39g，10mmol)的乙腈(40mL)溶液中缓慢加入N-溴代丁二酰亚胺(1.87g，10.5mmol)，继续搅拌1小时。反应液用乙酸乙酯萃取(50mL×3)，有机层依次用水、饱和食盐水洗涤，保留有机层，用无水硫酸钠干燥，过滤减压浓缩，粗产品经柱层析(石油醚：乙酸乙酯＝20：1)得2.9g无色透明油状物，产率为92％。Slow addition of N-bromosuccinyl to a solution of 6,7-dihydrothieno[3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl ester (2.39 g, 10 mmol) in acetonitrile (40 mL) The imine (1.87 g, 10.5 mmol) was stirred for an additional 1 hour. The reaction mixture was extracted with EtOAc (EtOAc (EtOAc)EtOAc. Ethyl ester = 20:1) 2.9 g of mp.
1H NMR(300MHz,Chloroform-d)δ6.74(s,1H),4.41(s,2H),3.69(t,J＝5.7Hz,2H),2.73(t,J＝5.8Hz,2H),1.48(s,9H)；MS(ESI):m/z 318[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ6.74 (s, 1H), 4.41 (s, 2H), 3.69 (t, J = 5.7Hz, 2H), 2.73 (t, J = 5.8Hz, 2H), 1.48 (s, 9H); MS (ESI): m/z 318 [M+H] + .
步骤3. 2-甲酰基-6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯：Step 3. 2-Formyl-6,7-dihydrothieno[3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000021
Figure PCTCN2016087368-appb-000021
-78℃，在氮气保护下，往2-溴-6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯(1g，3.75mmol)的无水四氢呋喃(30mL)溶液中缓慢滴加1.8mL正丁基锂的四氢呋喃溶液(2.5M)，继续搅拌1小时。往反应液中加入330mg无水N,N-二甲基甲酰胺，搅拌1小时后，加入1mL水淬灭。反应液用乙酸乙酯萃取(50mL×3)，有机层依次用水洗，饱和食盐水洗，保留有机层，用无水硫酸钠干燥，过滤减压浓缩，粗产品经柱层析(石油醚：乙酸乙酯＝10：1)得无色透明油状物841mg，产率为84％。To a residue of 2-bromo-6,7-dihydrothieno[3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl ester (1 g, 3.75 mmol) at -78 ° C under nitrogen. A solution of 1.8 mL of n-butyllithium in tetrahydrofuran (2.5 M) was slowly added dropwise to a solution of tetrahydrofuran (30 mL), and stirring was continued for one hour. 330 mg of anhydrous N,N-dimethylformamide was added to the reaction mixture, and the mixture was stirred for 1 hour, and then quenched with 1 mL of water. The reaction mixture was extracted with ethyl acetate (50 mL×3). EtOAc. Ethyl ester = 10:1) 841 mg of colorless transparent oil, yield 84%.
1H NMR(300MHz,Chloroform-d)δ9.82(s,1H),7.47(s,1H),4.52(s,2H),3.73(t,J＝5.5Hz,2H),2.91(t,J＝5.4Hz,2H),1.49(s,9H)；MS(ESI):m/z 268[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 9.82 (s, 1H), 7.47 (s, 1H), 4.52 (s, 2H), 3.73 (t, J = 5.5 Hz, 2H), 2.91 (t, J = 5.4 Hz, 2H), 1.49 (s, 9H); MS (ESI): m/z 268[M+H] + .
步骤4. 2-(4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-6,7-二氢噻吩并[3,2-c]吡啶 -5(4H)-羧酸叔丁酯：Step 4. 2-(4-(Methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-6,7-dihydrothieno[3,2-c]pyridine -5(4H)-tert-butyl carboxylic acid:
Figure PCTCN2016087368-appb-000022
Figure PCTCN2016087368-appb-000022
往2-甲酰基-6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯(830mg，3.1mmol)的乙醇(15mL)溶液中加入2,3-二氨基苯甲酸甲酯(540mg，3.1mmol)以及NaHSO3(810mg，7.8mmol)，加热至60℃，反应过夜。减压蒸去大部分乙醇，残留物用乙酸乙酯萃取(50mL×3)，有机层依次用水洗，饱和食盐水洗，保留有机层，用无水硫酸钠干燥，过滤减压浓缩，粗产品经柱层析(石油醚：乙酸乙酯＝4：1)得黄色粉末1.26g，产率为98％。Add 2,3 to a solution of 2-formyl-6,7-dihydrothieno[3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl ester (830 mg, 3.1 mmol) in ethanol (15 mL) - diamino benzoic acid methyl ester (540mg, 3.1mmol) and NaHSO 3 (810mg, 7.8mmol), was heated to 60 ℃, overnight. The organic layer was extracted with EtOAc (EtOAc) (EtOAc) Column chromatography (petroleum ether: ethyl acetate = 4:1) gave a white powder, 1.26 g, yield 98%.
1H NMR(300MHz,DMSO-d6)δ12.44(s,1H),8.04(s,1H),7.87(d,J＝7.8Hz,1H),7.80(d,J＝7.8Hz,1H),7.27(t,J＝7.8Hz,1H),4.47(s,2H),3.96(s,3H),3.66(t,J＝5.9Hz,2H),2.84(t,J＝5.9Hz,2H),1.42(s,9H)；MS(ESI)414[M+H]+。 1 H NMR (300MHz, DMSO- d 6) δ12.44 (s, 1H), 8.04 (s, 1H), 7.87 (d, J = 7.8Hz, 1H), 7.80 (d, J = 7.8Hz, 1H) , 7.27 (t, J = 7.8 Hz, 1H), 4.47 (s, 2H), 3.96 (s, 3H), 3.66 (t, J = 5.9 Hz, 2H), 2.84 (t, J = 5.9 Hz, 2H) , 1.42 (s, 9H); MS (ESI) 414 [M+H] + .
步骤5. 2-(4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 5. 2-(4,5,6,7-Tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxylic acid methyl ester:
Figure PCTCN2016087368-appb-000023
Figure PCTCN2016087368-appb-000023
将2-(4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯(4.6g，11.1mmol)溶于二氯甲烷(20mL)中，缓慢滴加三氟乙酸(5mL)，室温搅拌1小时。减压蒸去大部分溶剂，往残留物中加入饱和Na2CO3溶液(10mL)，用乙酸乙酯萃取(50mL×3)，有机层依次用水洗，饱和食盐水洗，保留有机层，用无水硫酸钠干燥，过滤减压浓缩，粗产品经柱层析(二氯甲烷：甲醇＝50：1)得黄色泡沫状固体3.0g，产率为87％。2-(4-(Methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-6,7-dihydrothieno[3,2-c]pyridine-5(4H)-carboxylate The acid tert-butyl ester (4.6 g, 11.1 mmol) was dissolved in dichloromethane (20 mL). Most of the solvent was distilled off under reduced pressure, to the residue was added saturated Na 2 CO 3 solution (10 mL), extracted with ethyl acetate (50mL × 3). The organic layer was washed with water and brine, the organic layer was retained, with no The organic layer was dried over sodium sulfate (MgSO4).
1H NMR(300MHz,DMSO-d6)δ12.63(brs,1H),9.24(brs,1H),8.07(s,1H),7.88(d,J＝7.9Hz,1H),7.81(d,J＝7.8Hz,1H),7.30(t,J＝7.8Hz,1H),4.27(s,2H),3.95(s,3H),3.47(t,J＝5.9Hz,2H),3.11(t,J＝5.4Hz,2H)；MS(ESI)314[M+H]+。 1 H NMR (300 MHz, DMSO-d 6 ) δ 12.63 (brs, 1H), 9.24 (brs, 1H), 8.07 (s, 1H), 7.78 (d, J = 7.9 Hz, 1H), 7.81 (d, J = 7.8 Hz, 1H), 7.30 (t, J = 7.8 Hz, 1H), 4.27 (s, 2H), 3.95 (s, 3H), 3.47 (t, J = 5.9 Hz, 2H), 3.11 (t, J = 5.4 Hz, 2H); MS (ESI) 314 [M+H] + .
步骤6. 2-(4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-1)：Step 6. 2-(4,5,6,7-Tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide (I-1) :
Figure PCTCN2016087368-appb-000024
Figure PCTCN2016087368-appb-000024
向2-(4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(110mg，0.35mmol)中加入氨气的饱和甲醇溶液(10mL)，80℃封管反应20小时。将反应液 冷却至室温，减压蒸去大部分溶剂，粗产品经柱层析(二氯甲烷：甲醇＝10：1)得白色粉末77mg，产率为74％。To methyl 2-(4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxylate (110 mg, 0.35 mmol) A saturated methanol solution of ammonia (10 mL) was added thereto, and the mixture was sealed at 80 ° C for 20 hours. Reaction solution After cooling to room temperature, most of the solvent was evaporated under reduced pressure, and the crude product was purified by column chromatography (dichloromethane:methanol = 10:1) to yield white powder (yield: 74%).
1H NMR(300MHz,DMSO-d6)δ9.09(brs,1H),7.83(dd,J＝7.6,1.1Hz,1H),7.73(brs,1H),7.68(dd,J＝8.0,1.1Hz,1H),7.63(s,1H),7.31(t,J＝7.8Hz,1H),3.81(s,2H),3.00(t,J＝5.6Hz,2H),2.76(t,J＝5.6Hz,2H)；13C NMR(125MHz,DMSO-d6)δ166.75,148.54,138.68,136.96,135.53,128.84,126.66,126.66,123.10,122.49,122.04,115.62,45.30,43.40,26.09；HRMS(ESI):m/z Calcd.For C15H15N4OS[M+H]+:299.0967；Found:299.0959。 1 H NMR (300MHz, DMSO- d 6) δ9.09 (brs, 1H), 7.83 (dd, J = 7.6,1.1Hz, 1H), 7.73 (brs, 1H), 7.68 (dd, J = 8.0,1.1 Hz, 1H), 7.63 (s, 1H), 7.31 (t, J = 7.8 Hz, 1H), 3.81 (s, 2H), 3.00 (t, J = 5.6 Hz, 2H), 2.76 (t, J = 5.6) Hz, 2H); 13 C NMR (125MHz, DMSO-d 6 ) δ 166.75, 148.54, 138.68, 136.96, 135.53, 128.84, 126.66, 126.66, 123.10, 122.49, 122.04, 115.62, 45.30, 43.40, 26.09; HRMS (ESI) : m/z Calcd. For C 15 H 15 N 4 OS [M+H] + : 299.0967; Found: 299.0959.
实施例2. 2-(5-甲基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-2的制备：Example 2. 2-(5-Methyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-methyl Preparation of amide, I-2:
步骤1. 2-(5-甲基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 1. 2-(5-Methyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxylic acid A ester:
Figure PCTCN2016087368-appb-000025
Figure PCTCN2016087368-appb-000025
将2-(4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(198mg，0.63mmol)溶于甲酸(3mL)中，加甲醛溶液(37％，0.3mL)。将反应液升至95℃下反应4小时至原料反应完全。减压蒸去大部分溶剂，往残留物中加入Na2CO3饱和水溶液调节pH至碱性，用乙酸乙酯萃取(50mL×3)，有机层依次用水、饱和食盐水洗涤，保留有机层，用无水硫酸钠干燥，过滤减压浓缩，粗产品经柱层析(二氯甲烷：甲醇＝80：1)得黄色泡固体128mg，产率为62％。Methyl 2-(4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxylate (198 mg, 0.63 mmol) Soluble in formic acid (3 mL), add formaldehyde solution (37%, 0.3 mL). The reaction solution was raised to 95 ° C for 4 hours until the starting material was completely reacted. Most of the solvent was distilled off under reduced pressure, to the residue was added Na 2 CO 3 saturated aqueous solution was adjusted to basic pH and extracted with ethyl acetate (50mL × 3), the organic layer was successively washed with water, saturated brine, the organic layer was retained, The organic layer was dried over anhydrous sodium sulfate (MgSO4).
1H NMR(300MHz,DMSO-d6)δ7.79(s,1H),7.77(d,J＝7.1Hz,1H),7.69(d,J＝7.0Hz,1H),7.16(t,J＝7.5Hz,1H),3.91(s,3H),3.42(s,2H),2.84(t,J＝5.9Hz,2H),2.66(t,J＝5.9Hz,2H),2.36(s,3H)；MS(ESI)328[M+H]+。 1 H NMR (300MHz, DMSO- d 6) δ7.79 (s, 1H), 7.77 (d, J = 7.1Hz, 1H), 7.69 (d, J = 7.0Hz, 1H), 7.16 (t, J = 7.5 Hz, 1H), 3.91 (s, 3H), 3.42 (s, 2H), 2.84 (t, J = 5.9 Hz, 2H), 2.66 (t, J = 5.9 Hz, 2H), 2.36 (s, 3H) ;MS (ESI) 328 [M+H] + .
步骤2. 2-(5-甲基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-2)Step 2. 2-(5-Methyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide (I-2)
Figure PCTCN2016087368-appb-000026
Figure PCTCN2016087368-appb-000026
按照制备化合物I-1的操作方法，以2-(5-甲基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(128mg，0.39mmol)为原料氨解得到白色粉末94mg，产率为77％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 2-(5-methyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[ d] Methyl imidazole-4-carboxylate (128 mg, 0.39 mmol) was obtained as a crude material to give a white powder (yield: 77%, yield: 77% (dichloromethane:methanol = 10:1).
1H NMR(300MHz,DMSO-d6)δ9.49(brs,1H),7.74(dd,J＝7.5,1.2Hz,1H),7.65 (dd,J＝7.9,1.2Hz,1H),7.59(brs,1H),7.57(s,1H),7.18(t,J＝7.7Hz,1H),3.47(s,2H),2.90(t,J＝5.6Hz,2H),2.71(t,J＝5.6Hz,2H),2.41(s,3H)；13C NMR(125MHz,DMSO-d6)δ167.58,151.40,143.15,140.55,135.75,135.59,133.04,125.06,121.69,121.20,120.59,116.90,54.71,52.43,45.66,25.74；HRMS(ESI):m/z Calcd.For C16H17N4OS[M+H]+:313.1123；Found:313.1114。 1 H NMR (300MHz, DMSO- d 6) δ9.49 (brs, 1H), 7.74 (dd, J = 7.5,1.2Hz, 1H), 7.65 (dd, J = 7.9,1.2Hz, 1H), 7.59 ( Brs, 1H), 7.57 (s, 1H), 7.18 (t, J = 7.7 Hz, 1H), 3.47 (s, 2H), 2.90 (t, J = 5.6 Hz, 2H), 2.71 (t, J = 5.6 Hz, 2H), 2.41 (s, 3H); 13 C NMR (125MHz, DMSO-d 6 ) δ 167.58, 151.40, 143.15, 140.55, 135.75, 135.59, 133.04, 125.06, 121.69, 121.20, 120.59, 116.90, 54.71, 52.43 , 45.66, 25.74; HRMS (ESI): m/z Calcd. For C 16 H 17 N 4 OS [M+H] + : 313.1123; Found: 313.1114.
实施例3. 2-(5-乙基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-3的制备：Example 3. 2-(5-Ethyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-methyl Preparation of amide, I-3:
步骤1. 2-(5-乙基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 1. 2-(5-Ethyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxylic acid A ester:
Figure PCTCN2016087368-appb-000027
Figure PCTCN2016087368-appb-000027
将2-(4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(160mg，0.51mmol)溶于无水DMF(5mL)中，加入Cs2CO3(183mg，0.56mmol)，溴乙烷(67mg，0.61mmol)后，升温至80℃下反应1小时。将反应液倒入水中，乙酸乙酯萃取(50mL×3)，有机层依次用水洗，饱和食盐水洗，保留有机层，用无水硫酸钠干燥，过滤减压浓缩，粗产品经柱层析(二氯甲烷：甲醇＝80：1)得浅黄色胶状物150mg，产率为86％。Methyl 2-(4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxylate (160 mg, 0.51 mmol) After dissolving in anhydrous DMF (5 mL), Cs 2 CO 3 (183 mg, 0.56 mmol), bromoethane (67 mg, 0.61 mmol), and the mixture was heated to 80 ° C for 1 hour. The reaction mixture was poured into water and extracted with ethyl acetate (50 mL×3). Dichloromethane:methanol = 80:1) gave a pale yellow gum, 150 mg, yield 86%.
1H NMR(300MHz,DMSO-d6)δ12.42(brs,1H),7.97(s,1H),7.86(d,J＝7.9Hz,1H),7.78(d,J＝7.7Hz,1H),7.28(t,J＝7.8Hz,1H),3.96(s,3H),3.54(s,2H),2.86(brs,2H),2.77(brs,2H),2.58(q,J＝6.9Hz,2H),1.10(t,J＝7.1Hz,3H)；MS(ESI)342[M+H]+。 1 H NMR (300MHz, DMSO- d 6) δ12.42 (brs, 1H), 7.97 (s, 1H), 7.86 (d, J = 7.9Hz, 1H), 7.78 (d, J = 7.7Hz, 1H) , 7.28 (t, J = 7.8 Hz, 1H), 3.96 (s, 3H), 3.54 (s, 2H), 2.86 (brs, 2H), 2.77 (brs, 2H), 2.58 (q, J = 6.9 Hz, 2H), 1.10 (t, J = 7.1 Hz, 3H); MS (ESI) 342 [M+H] + .
步骤2. 2-(5-乙基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-3)：Step 2. 2-(5-Ethyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide (I-3):
Figure PCTCN2016087368-appb-000028
Figure PCTCN2016087368-appb-000028
按照制备化合物I-1的操作方法，以2-(5-乙基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(150mg，0.44mmol)为原料氨解得到浅黄色固体87mg，产率为61％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 2-(5-ethyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[ d] Methyl imidazole-4-carboxylate (150 mg, 0.44 mmol) was obtained as a crude material (yield: 87 mg,yield: 61% (dichloromethane:methanol = 10:1).
1H NMR(500MHz,DMSO-d6)δ13.44(brs,1H),9.14(brs,1H),7.84(d,J＝7.6Hz,1H),7.78(brs,1H),7.68(d,J＝8.0Hz,1H),7.64(s,1H),7.32(t,J＝7.7Hz,1H),3.66(s,2H),2.92(s,2H),2.89(brs,2H),2.67(brs,2H),1.14(t,J＝7.2Hz,3H)；13C NMR (125MHz,DMSO-d6)δ166.02,147.64,141.29,137.31,135.04,129.40,126.13,122.83,122.23,121.93,114.62,51.36,50.76,49.46,24.89,11.86；HRMS(ESI):m/z Calcd.For C17H19N4OS[M+H]+:327.1280；Found:327.1273。 1 H NMR (500MHz, DMSO- d 6) δ13.44 (brs, 1H), 9.14 (brs, 1H), 7.84 (d, J = 7.6Hz, 1H), 7.78 (brs, 1H), 7.68 (d, J = 8.0 Hz, 1H), 7.64 (s, 1H), 7.32 (t, J = 7.7 Hz, 1H), 3.66 (s, 2H), 2.92 (s, 2H), 2.89 (brs, 2H), 2.67 ( Brs, 2H), 1.14 (t, J = 7.2 Hz, 3H); 13 C NMR (125MHz, DMSO-d 6 ) δ 166.02, 147.64, 141.29, 137.31, 135.04, 129.40, 126.13, 122.83, 122.23, 121.93, 114.62, 51.36, 50.76, 49.46, 24.89, 11.86; HRMS (ESI): m/z Calcd. For C 17 H 19 N 4 OS [M+H] + : 327.1280; Found: 327.1273.
实施例4. 2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-4的制备：Example 4. 2-(4,5,6,7-Tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide, I-4 Preparation:
步骤1. 4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 1. 4,7-Dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000029
Figure PCTCN2016087368-appb-000029
按照实施例1中步骤1的操作方法，以4,5,6,7-四氢噻吩[2,3-c]吡啶盐酸盐(1.00g，5.7mmol)为原料经保护得白色固体1.32g，产率为97％(石油醚：乙酸乙酯＝20：1)。According to the procedure of Step 1 in Example 1, 4,5,6,7-tetrahydrothiophene [2,3-c]pyridine hydrochloride (1.00 g, 5.7 mmol) was used as a material to afford white solid 1.32 g. The yield was 97% (petroleum ether: ethyl acetate = 20:1).
1H NMR(300MHz,Chloroform-d)δ7.13(d,J＝5.0Hz,1H),6.79(d,J＝5.1Hz,1H),4.62(s,2H),3.67(t,J＝5.4Hz,2H),2.70(t,J＝5.4Hz,2H),1.8(s,9H)；MS(ESI)240[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ7.13 (d, J = 5.0Hz, 1H), 6.79 (d, J = 5.1Hz, 1H), 4.62 (s, 2H), 3.67 (t, J = 5.4 Hz, 2H), 2.70 (t, J = 5.4 Hz, 2H), 1.8 (s, 9H); MS (ESI) 240 [M+H] + .
步骤2. 2-溴-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 2. 2-Bromo-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000030
Figure PCTCN2016087368-appb-000030
按照实施例1中步骤2的操作方法，以4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(1.32g，5.5mmol)为原料经溴代得无色透明油状物1.65g，产率为94％(石油醚：乙酸乙酯＝20：1)。Bromine according to the procedure of Step 2 in Example 1, using 4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (1.32 g, 5.5 mmol) as the starting material A colorless transparent oil of 1.65 g was obtained in a yield of 94% ( petroleum ether: ethyl acetate = 20:1).
1H NMR(300MHz,Chloroform-d)δ6.74(s,1H),4.50(s,2H),3.64(t,J＝5.7Hz,2H),2.63(t,J＝5.8Hz,2H),1.47(s,9H)；MS(ESI):m/z 318[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ6.74 (s, 1H), 4.50 (s, 2H), 3.64 (t, J = 5.7Hz, 2H), 2.63 (t, J = 5.8Hz, 2H), 1.47 (s, 9H); MS (ESI): m/z 318 [M+H] + .
步骤3. 2-甲酰基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 3. 2-Formyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000031
Figure PCTCN2016087368-appb-000031
按照实施例1中步骤3的操作方法，以2-溴-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(1.65g，5.2mmol)为原料制备得到浅棕色胶状物1.12g，产率为81％(石油醚：乙酸乙酯＝10：1)。Following the procedure of Step 3 in Example 1, 2-bromo-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (1.65 g, 5.2 mmol) 1.12 g of a light brown gum was obtained as a starting material, yield 81% ( petroleum ether: ethyl acetate = 10:1).
1H NMR(300MHz,Chloroform-d)δ9.82(s,1H),7.47(s,1H),4.67(s,2H),3.68(t,J＝5.6Hz,2H),2.75(d,J＝5.3Hz,2H),1.47(s,9H)；MS(ESI):m/z 268[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 9.82 (s, 1H), 7.47 (s, 1H), 4.67 (s, 2H), 3.68 (t, J = 5.6 Hz, 2H), 2.75 (d, J) = 5.3 Hz, 2H), 1.47 (s, 9H); MS (ESI): m/z 268 [M+H] + .
步骤4. 2-(4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 4. 2-(4-(Methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7-dihydrothieno[2,3-c]pyridine-6(5H) - tert-butyl carboxylate:
Figure PCTCN2016087368-appb-000032
Figure PCTCN2016087368-appb-000032
按照实施例1中步骤4的操作方法，以2-甲酰基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(1.12g，4.2mmol)和2,3-二氨基苯甲酸甲酯(0.70g，4.2mmol)为原料 制备得到黄色粉末1.53g，产率为88％(石油醚：乙酸乙酯＝3：1)。According to the procedure of Step 4 in Example 1, 2-formyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (1.12 g, 4.2 mmol) And 2,3-diaminobenzoic acid methyl ester (0.70 g, 4.2 mmol) as raw materials A yellow powder of 1.53 g was obtained in a yield of 88% (petrole ether: ethyl acetate = 3:1).
1H NMR(300MHz,Chloroform-d)δ10.46(brs,1H),7.95(d,J＝7.8Hz,1H),7.87(d,J＝7.7Hz,1H),7.37(s,1H),7.29(t,J＝7.8Hz,1H),4.68(brs,2H),4.01(s,3H),3.73(t,J＝5.6Hz,2H),2.76(t,J＝5.6Hz,2H),1.50(s,9H)；MS(ESI)414[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ10.46 (brs, 1H), 7.95 (d, J = 7.8Hz, 1H), 7.87 (d, J = 7.7Hz, 1H), 7.37 (s, 1H), 7.29 (t, J = 7.8 Hz, 1H), 4.68 (brs, 2H), 4.01 (s, 3H), 3.73 (t, J = 5.6 Hz, 2H), 2.76 (t, J = 5.6 Hz, 2H), 1.50 (s, 9H); MS (ESI) 414 [M+H] + .
步骤5. 2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 5. 2-(4,5,6,7-Tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxylic acid methyl ester:
Figure PCTCN2016087368-appb-000033
Figure PCTCN2016087368-appb-000033
按照实施例1中步骤5的操作方法，以2-(4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(1.53g，3.7mmol)为原料去保护基得到浅黄色粉末0.93g，产率为80％(二氯甲烷：甲醇＝50：1)。Following the procedure of Step 5 in Example 1, 2-(4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7-dihydrothieno[2,3 -c] pyridine-6(5H)-carboxylic acid tert-butyl ester (1.53 g, 3.7 mmol) as a starting material deprotecting group afforded a pale yellow powder (yield:::::::::::::::: .
1H NMR(300MHz,Chloroform-d)δ10.46(brs,1H),7.94(d,J＝8.0Hz,1H),7.85(d,J＝7.8Hz,1H),7.37(s,1H),7.28(t,J＝7.9Hz,1H),4.08(brs,2H),4.00(s,3H),3.15(t,J＝5.8Hz,2H),2.69(t,J＝5.8Hz,2H)；MS(ESI)314[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ10.46 (brs, 1H), 7.94 (d, J = 8.0Hz, 1H), 7.85 (d, J = 7.8Hz, 1H), 7.37 (s, 1H), 7.28 (t, J = 7.9 Hz, 1H), 4.08 (brs, 2H), 4.00 (s, 3H), 3.15 (t, J = 5.8 Hz, 2H), 2.69 (t, J = 5.8 Hz, 2H); MS (ESI) 314 [M + H] + .
步骤6. 2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-4)：Step 6. 2-(4,5,6,7-Tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide (I-4) :
Figure PCTCN2016087368-appb-000034
Figure PCTCN2016087368-appb-000034
按照制备化合物I-1的操作方法，以2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(158mg，0.50mmol)为原料氨解得到白色固体130mg，产率为87％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4 Methyl formate (158 mg, 0.50 mmol) was obtained as a crude material to give a white solid (yield: <RTIgt;</RTI>
1H NMR(500MHz,DMSO-d6)δ9.12(brs,1H),7.82(d,J＝7.5Hz,1H),7.75(s,1H),7.69(dd,J＝7.9,0.9Hz,1H),7.66(brs,1H),7.31(t,J＝7.8Hz,1H),3.92(s,2H),2.96(t,J＝5.7Hz,2H),2.62(t,J＝5.4Hz,2H)；13C NMR(125MHz,DMSO-d6)δ166.71,148.52,141.84,139.75,136.01,128.92,123.15,122.53,115.26,44.83,42.98,26.57；HRMS(ESI):m/z Calcd.For C15H15N4OS[M+H]+:299.0967；Found:299.0959。 1 H NMR (500MHz, DMSO- d 6) δ9.12 (brs, 1H), 7.82 (d, J = 7.5Hz, 1H), 7.75 (s, 1H), 7.69 (dd, J = 7.9,0.9Hz, 1H), 7.66 (brs, 1H), 7.31 (t, J = 7.8 Hz, 1H), 3.92 (s, 2H), 2.96 (t, J = 5.7 Hz, 2H), 2.62 (t, J = 5.4 Hz, 2H); 13 C NMR (125MHz, DMSO-d 6 ) δ 166.71, 148.52, 141.84, 139.75, 136.01, 128.92, 123.15, 122.53, 115.26, 44.83, 42.98, 26.57; HRMS (ESI): m/z Calcd.For C 15 H 15 N 4 OS [M+H] + : 299.0967; Found: 299.0959.
实施例5. 2-(6-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-5的制备：Example 5. 2-(6-Methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-methyl Preparation of amide, I-5:
步骤1. 2-(6-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯： Step 1. 2-(6-Methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxylic acid A Ester:
Figure PCTCN2016087368-appb-000035
Figure PCTCN2016087368-appb-000035
按照实施例2中步骤1的操作方法，以2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(158mg，0.50mmol)为原料，经甲基化得棕黄色泡沫状固体134mg，产率为82％(二氯甲烷：甲醇＝30：1)。Following the procedure of Step 1 in Example 2, 2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole- Methyl 4-formic acid (158 mg, 0.50 mmol) was obtained from EtOAc (EtOAc: EtOAc: EtOAc:
1H NMR(400MHz,Chloroform-d)δ10.47(brs,1H),7.96(d,J＝7.9Hz,1H),7.88(d,J＝7.7Hz,1H),7.41(s,1H),7.31(t,J＝7.8Hz,1H),4.04(s,3H),3.71(s,2H),2.87-2.82(m,2H),2.81-2.76(m,2H),2.54(s,3H)；MS(ESI)328[M+H]+。 1 H NMR (400MHz, Chloroform- d) δ10.47 (brs, 1H), 7.96 (d, J = 7.9Hz, 1H), 7.88 (d, J = 7.7Hz, 1H), 7.41 (s, 1H), 7.31 (t, J = 7.8 Hz, 1H), 4.04 (s, 3H), 3.71 (s, 2H), 2.87-2.82 (m, 2H), 2.81-2.76 (m, 2H), 2.54 (s, 3H) ;MS (ESI) 328 [M+H] + .
步骤2. 2-(6-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-5)：Step 2. 2-(6-Methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide (I-5):
Figure PCTCN2016087368-appb-000036
Figure PCTCN2016087368-appb-000036
按照制备化合物I-1的操作方法，以2-(6-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-羧酸甲酯(134mg，0.41mmol)为原料氨解得到灰白色固体102mg，产率为80％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 2-(6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[ d] Methyl imidazole-4-carboxylate (134 mg, 0.41 mmol) was obtained as a crude material (yield: EtOAc (EtOAc)
1H NMR(500MHz,DMSO-d6)δ9.15(brs,1H),7.84(d,J＝6.3Hz,1H),7.77(brs,1H),7.69(d,J＝7.9Hz,1H),7.65(brs,1H),7.32(t,J＝7.4Hz,1H),3.60(s,2H),2.73(t,J＝5.0Hz,2H),2.67(t,J＝5.5Hz,2H),2.39(s,3H)；13C NMR(125MHz,DMSO-d6)δ166.54,148.28,141.82,137.62,135.55,135.14,129.45,128.26,123.31,122.71,122.43,115.12,53.74,52.01,45.42,25.67；HRMS(ESI):m/z Calcd.For C16H17N4OS[M+H]+:313.1123；Found:313.1118。 1 H NMR (500MHz, DMSO- d 6) δ9.15 (brs, 1H), 7.84 (d, J = 6.3Hz, 1H), 7.77 (brs, 1H), 7.69 (d, J = 7.9Hz, 1H) , 7.65 (brs, 1H), 7.32 (t, J = 7.4 Hz, 1H), 3.60 (s, 2H), 2.73 (t, J = 5.0 Hz, 2H), 2.67 (t, J = 5.5 Hz, 2H) , 2.39(s,3H); 13 C NMR (125MHz, DMSO-d 6 ) δ 166.54, 148.28, 141.82, 137.62, 135.55, 135.14, 129.45, 128.26, 123.31, 122.71, 122.43, 115.12, 53.74, 52.01, 45.42, 25.67 HRMS (ESI): m/z Calcd. for C 16 H 17 N 4 OS [M+H] + : 313.1123; Found: 313.1118.
实施例6. 6-氯-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-6的制备：Example 6. 6-Chloro-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide , Preparation of I-6:
步骤1. 5-氯-7-硝基二氢吲哚-2,3-二酮：Step 1. 5-Chloro-7-nitroindoline-2,3-dione:
Figure PCTCN2016087368-appb-000037
Figure PCTCN2016087368-appb-000037
-5℃下，向溶有5-氯靛红(1.81g，10mmol)的浓硫酸(6mL)溶液中缓慢滴加发烟硝酸(1mL)，搅拌1小时，反应液倒入冰水中，析出固体，过滤水洗，干燥得黄色固体1.77g，产率为78％。To a solution of 5-chloroindole (1.81 g, 10 mmol) in concentrated sulfuric acid (6 mL) was slowly added dropwise fuming nitric acid (1 mL) at -5 ° C, stirred for 1 hour, and poured into ice water to precipitate solids. It was filtered, washed with water and dried to give a white solid.
1H NMR(300MHz,DMSO-d6)δ11.80(s,1H),8.32(d,J＝2.2Hz,1H),8.02(d,J＝ 2.3Hz,1H),4.48(s,1H)；MS(ESI)227[M+H]+。 1 H NMR (300MHz, DMSO- d 6) δ11.80 (s, 1H), 8.32 (d, J = 2.2Hz, 1H), 8.02 (d, J = 2.3Hz, 1H), 4.48 (s, 1H) ;MS (ESI) 227 [M+H] + .
步骤2. 2-氨基-5-氯-3-硝基苯甲酸：Step 2. 2-Amino-5-chloro-3-nitrobenzoic acid:
Figure PCTCN2016087368-appb-000038
Figure PCTCN2016087368-appb-000038
0℃下，将5-氯-7-硝基二氢吲哚-2,3-二酮(1.77g，7.8mmol)加至氢氧化钠水溶液(5N，15mL)中形成混悬液，再往混悬液中缓慢滴加30％过氧化氢溶液(2mL)，滴加完毕后移至室温继续搅拌5小时，用2N盐酸中和至pH＝4.0，析出沉淀过滤，水洗，干燥得黄色固体1.17g，产率为69％。Add 5-chloro-7-nitroindoline-2,3-dione (1.77 g, 7.8 mmol) to aqueous sodium hydroxide (5N, 15 mL) at 0 ° C to form a suspension. 30% hydrogen peroxide solution (2mL) was slowly added dropwise to the suspension. After the addition was completed, it was moved to room temperature and stirred for 5 hours. It was neutralized with 2N hydrochloric acid to pH=4.0, and the precipitate was precipitated, washed with water and dried to give a yellow solid. g, yield was 69%.
1H NMR(300MHz,DMSO-d6)δ13.92(brs,1H),8.46(brs,2H),8.28(d,J＝2.7Hz,1H),8.11(d,J＝2.6Hz,1H)；MS(ESI)217[M+H]+。 1 H NMR (300MHz, DMSO- d 6) δ13.92 (brs, 1H), 8.46 (brs, 2H), 8.28 (d, J = 2.7Hz, 1H), 8.11 (d, J = 2.6Hz, 1H) ;MS (ESI) 217 [M+H] + .
步骤3. 2-氨基-5-氯-3-硝基苯甲酸甲酯：Step 3. Methyl 2-amino-5-chloro-3-nitrobenzoate:
Figure PCTCN2016087368-appb-000039
Figure PCTCN2016087368-appb-000039
往2-氨基-5-氯-3-硝基苯甲酸(1.17g，5.38mmol)的甲醇(60mL)溶液中加入一水合对甲苯磺酸(1.02g，5.38mmol)，加热回流36小时。冷却反应液，减压蒸除溶剂，粗产品经柱层析(石油醚：乙酸乙酯＝8：1)得黄色固体0.53g，产率为43％。To a solution of 2-amino-5-chloro-3-nitrobenzoic acid (1.17 g, 5.38 mmol) in MeOH (60 mL), EtOAc. The reaction mixture was cooled, and the solvent was evaporated evaporated evaporated.
1H NMR(300MHz,Chloroform-d)δ8.43(brs,2H),8.38(d,J＝2.6Hz,1H),8.20(d,J＝2.6Hz,1H),3.93(s,3H)；MS(ESI)231[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ8.43 (brs, 2H), 8.38 (d, J = 2.6Hz, 1H), 8.20 (d, J = 2.6Hz, 1H), 3.93 (s, 3H); MS (ESI) 231 [M + H] + .
步骤4. 2,3-二氨基-5-氯苯甲酸甲酯：Step 4. Methyl 2,3-diamino-5-chlorobenzoate:
Figure PCTCN2016087368-appb-000040
Figure PCTCN2016087368-appb-000040
往2-氨基-5-氯-3-硝基苯甲酸甲酯(530mg，2.31mmol)的甲醇(30mL)溶液中加入10％的钯炭50mg，置换氮气三次，再置换氢气三次，在一个大气压的氢气氛围下搅拌5小时，过滤，减压蒸去大部分溶剂，粗产品经柱层析(石油醚：乙酸乙酯＝4：1)得灰白色固体51mg，产率为11％。To a solution of methyl 2-amino-5-chloro-3-nitrobenzoate (530 mg, 2.31 mmol) in methanol (30 mL), 50% of 10% palladium charcoal, replaced with nitrogen three times, and then replaced hydrogen three times at one atmosphere The mixture was stirred for 5 hours under a hydrogen atmosphere, filtered, and evaporated to dryness, and the solvent was evaporated to dryness (yield: ethyl ether: ethyl acetate = 4:1).
1H NMR(300MHz,Chloroform-d)δ7.27(d,J＝1.9Hz,1H),6.70(d,J＝1.9Hz,1H),4.46(brs,4H),3.86(s,3H),2.21(s,3H)；MS(ESI)201[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ7.27 (d, J = 1.9Hz, 1H), 6.70 (d, J = 1.9Hz, 1H), 4.46 (brs, 4H), 3.86 (s, 3H), 2.21 (s, 3H); MS (ESI) 201 [M+H] + .
步骤5. 2-(6-氯-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯： Step 5. 2-(6-Chloro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7-dihydrothieno[2,3-c]pyridine- 6(5H)-tert-butyl carboxylic acid:
Figure PCTCN2016087368-appb-000041
Figure PCTCN2016087368-appb-000041
按照实施例1中步骤4的操作方法，以2,3-二氨基-5-氯苯甲酸甲酯(51mg，0.25mmol)和2-甲酰基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(67mg，0.25mmol)为原料制备得到淡黄色粉末73mg，产率为65％(石油醚：乙酸乙酯＝3：1)。Following the procedure of Step 4 in Example 1, methyl 2,3-diamino-5-chlorobenzoate (51 mg, 0.25 mmol) and 2-formyl-4,7-dihydrothieno[2,3 -c] Pyridyl-6(5H)-carboxylic acid tert-butyl ester (67 mg, 0.25 mmol) was obtained as a crude material (yield: 73 mg,yield: 65% (ethyl ether: ethyl acetate = 3:1).
1H NMR(300MHz,Chloroform-d)δ10.49(brs,1H),7.88(d,J＝1.8Hz,1H),7.82(d,J＝1.7Hz,1H),7.38(s,1H),4.67(s,2H),4.00(s,3H),3.71(t,J＝5.0Hz,2H),2.75(t,J＝5.3Hz,2H),1.50(s,9H)；MS(ESI)448[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ10.49 (brs, 1H), 7.88 (d, J = 1.8Hz, 1H), 7.82 (d, J = 1.7Hz, 1H), 7.38 (s, 1H), 4.67 (s, 2H), 4.00 (s, 3H), 3.71 (t, J = 5.0 Hz, 2H), 2.75 (t, J = 5.3 Hz, 2H), 1.50 (s, 9H); MS (ESI) 448 [M+H] + .
步骤6. 6-氯-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 6. 6-Chloro-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxylic acid methyl ester :
Figure PCTCN2016087368-appb-000042
Figure PCTCN2016087368-appb-000042
按照实施例1中步骤5的操作方法，以2-(6-氯-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(73mg，0.16mmol)为原料去保护基得到淡棕色粉末48mg，产率为86％(二氯甲烷：甲醇＝50：1)。Following the procedure of Step 5 in Example 1, 2-(6-chloro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7-dihydrothiophene [2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (73 mg, 0.16 mmol) was obtained as a starting material to afford the title compound as a pale brown powder (yield: 86 mg, yield: 86%) 1).
1H NMR(300MHz,Chloroform-d)δ10.50(brs,1H),7.89(d,J＝1.9Hz,1H),7.82(d,J＝1.9Hz,1H),7.41(s,1H),6.86(s,1H),4.10(s,2H),4.02(s,3H),3.16(t,J＝5.8Hz,2H),2.72(t,J＝5.6Hz,2H)；MS(ESI)348[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ10.50 (brs, 1H), 7.89 (d, J = 1.9Hz, 1H), 7.82 (d, J = 1.9Hz, 1H), 7.41 (s, 1H), 6.86 (s, 1H), 4.10 (s, 2H), 4.02 (s, 3H), 3.16 (t, J = 5.8 Hz, 2H), 2.72 (t, J = 5.6 Hz, 2H); MS (ESI) 348 [M+H] + .
步骤7. 6-氯-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-6)：Step 7. 6-Chloro-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide ( I-6):
Figure PCTCN2016087368-appb-000043
Figure PCTCN2016087368-appb-000043
按照制备化合物I-1的操作方法，以6-氯-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(48mg，0.14mmol)为原料氨解得到淡棕色固体30mg，产率为65％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 6-chloro-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d The imidazole-4-carboxylic acid methyl ester (48 mg, 0.14 mmol) was obtained as a crude material to give 30% of pale brown solid (yield: methylene chloride:methanol = 10:1).
1H NMR(500MHz,DMSO-d6)δ8.96(brs,1H),7.94(brs,1H),7.83(s,1H),7.74(d,J＝2.2Hz,1H)7.73(d,J＝2.2Hz,1H),4.13(s,2H),3.15(t,J＝6.2Hz,2H),2.76(t,J＝6.2Hz,2H)；13C NMR(125MHz,DMSO-d6)δ165.49,149.53,136.28,135.26,129.80,129.21,126.88,122.73,115.09,43.52,41.95,29.47；HRMS(ESI):m/z Calcd.For C15H14ClN4OS[M+H]+:333.0577；Found:333.0570。 1 H NMR (500MHz, DMSO- d 6) δ8.96 (brs, 1H), 7.94 (brs, 1H), 7.83 (s, 1H), 7.74 (d, J = 2.2Hz, 1H) 7.73 (d, J =2.2 Hz, 1H), 4.13 (s, 2H), 3.15 (t, J = 6.2 Hz, 2H), 2.76 (t, J = 6.2 Hz, 2H); 13 C NMR (125 MHz, DMSO-d 6 ) δ 165 .49,149.53,136.28,135.26,129.80,129.21,126.88,122.73,115.09,43.52,41.95,29.47;HRMS(ESI): m/z Calcd.For C 15 H 14 ClN 4 OS[M+H] + :333.0577; Found: 333.0570.
实施例7. 6-甲基-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰 胺，I-7的制备：Example 7. 6-Methyl-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-methyl Acyl Preparation of amines, I-7:
步骤1. 5-甲基-7-硝基二氢吲哚-2,3-二酮：Step 1. 5-Methyl-7-nitroindoline-2,3-dione:
Figure PCTCN2016087368-appb-000044
Figure PCTCN2016087368-appb-000044
按照实施例6中步骤1的操作方法，以5-甲基靛红(2.49g，20mmol)为原料，硝化制得黄色固体1.50g，产率为37％。According to the procedure of Step 1 in Example 6, 5-methyl-ruthenium (2.49 g, 20 mmol) was used as a starting material to obtain a yellow solid 1.50 g, yield 37%.
1H NMR(300MHz,DMSO-d6)δ11.58(brs,1H),8.13(s,1H),7.77(s,1H),2.35(s,3H)；MS(ESI)207[M+H]+。 1 H NMR (300 MHz, DMSO-d 6 ) δ 11.58 (brs, 1H), 8.13 (s, 1H), 7.77 (s, 1H), 2.35 (s, 3H); MS (ESI) 207 [M+H ] + .
步骤2. 2-氨基-5-甲基-3-硝基苯甲酸：Step 2. 2-Amino-5-methyl-3-nitrobenzoic acid:
Figure PCTCN2016087368-appb-000045
Figure PCTCN2016087368-appb-000045
按照实施例6中步骤2的操作方法，以5-甲基-7-硝基二氢吲哚-2,3-二酮(1.5g，7.4mmol)为原料制得黄色固体1.23g，产率为85％。Prepare a yellow solid 1.23 g of 5-methyl-7-nitroindoline-2,3-dione (1.5 g, 7.4 mmol) according to the procedure of Step 2 in Example 6. It is 85%.
1H NMR(300MHz,DMSO-d6)δ13.50(brs,1H),8.33(brs,2H),8.14(s,1H),8.07(s,1H),2.24(s,3H)；MS(ESI)197[M+H]+。 1 H NMR (300MHz, DMSO- d 6) δ13.50 (brs, 1H), 8.33 (brs, 2H), 8.14 (s, 1H), 8.07 (s, 1H), 2.24 (s, 3H); MS ( ESI) 197 [M+H] + .
步骤3. 2-氨基-5-甲基-3-硝基苯甲酸甲酯：Step 3. Methyl 2-amino-5-methyl-3-nitrobenzoate:
Figure PCTCN2016087368-appb-000046
Figure PCTCN2016087368-appb-000046
按照实施例6中步骤3的操作方法，以2-氨基-5-甲基-3-硝基苯甲酸(1.23g，6.3mmol)为原料制得黄色固体0.50g，产率为38％(石油醚：乙酸乙酯＝8：1)。According to the procedure of Step 3 in Example 6, 2-amino-5-methyl-3-nitrobenzoic acid (1.23 g, 6.3 mmol) was used as a starting material to obtain 0.50 g of a yellow solid. Ether: ethyl acetate = 8:1).
1H NMR(300MHz,Chloroform-d)δ8.21(s,1H),8.08(s,1H),7.71(brs,2H),3.91(s,3H),2.29(s,3H)；MS(ESI)211[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ8.21 (s, 1H), 8.08 (s, 1H), 7.71 (brs, 2H), 3.91 (s, 3H), 2.29 (s, 3H); MS (ESI ) 211 [M+H] + .
步骤4. 2,3-二氨基-5-甲基苯甲酸甲酯：Step 4. Methyl 2,3-diamino-5-methylbenzoate:
Figure PCTCN2016087368-appb-000047
Figure PCTCN2016087368-appb-000047
按照实施例6中步骤4的操作方法，以2-氨基-5-甲基-3-硝基苯甲酸甲酯(500mg，2.4mmol)为原料制得棕色固体90mg，产率为21％(石油醚：乙酸乙酯＝4：1)。According to the procedure of Step 4 in Example 6, a brown solid (yield: <RTI ID=0.0>> Ether: ethyl acetate = 4:1).
1H NMR(300MHz,Chloroform-d)δ7.27(d,J＝1.9Hz,1H),6.70(d,J＝1.9Hz,1H),4.46(brs,4H),3.86(s,3H),2.21(s,3H)；MS(ESI)181[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ7.27 (d, J = 1.9Hz, 1H), 6.70 (d, J = 1.9Hz, 1H), 4.46 (brs, 4H), 3.86 (s, 3H), 2.21 (s, 3H); MS (ESI) 181 [M+H] + .
步骤5. 2-(4-(甲氧基羰基)-6-甲基-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡 啶-6(5H)-羧酸叔丁酯：Step 5. 2-(4-(Methoxycarbonyl)-6-methyl-1H-benzo[d]imidazol-2-yl)-4,7-dihydrothieno[2,3-c]pyridin Pyridin-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000048
Figure PCTCN2016087368-appb-000048
按照实施例1中步骤4的操作方法，以2,3-二氨基-5-甲基苯甲酸甲酯(90mg，0.5mmol)和2-甲酰基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(134mg，0.5mmol)为原料制备得到淡黄色粉末96mg，产率为45％(石油醚：乙酸乙酯＝3：1)。Following the procedure of Step 4 in Example 1, methyl 2,3-diamino-5-methylbenzoate (90 mg, 0.5 mmol) and 2-formyl-4,7-dihydrothiophene [2, 3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (134 mg, 0.5 mmol) was obtained as a crude material (yield: EtOAc, EtOAc: EtOAc:
1H NMR(300MHz,Chloroform-d)δ10.40(brs,1H),7.72(s,1H),7.69(s,1H),7.35(s,1H),4.66(s,2H),3.98(s,3H),3.70(t,J＝5.3Hz,2H),2.73(t,J＝4.9Hz,2H),2.48(s,3H),1.49(s,9H)；MS(ESI)428[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ10.40 (brs, 1H), 7.72 (s, 1H), 7.69 (s, 1H), 7.35 (s, 1H), 4.66 (s, 2H), 3.98 (s , 3H), 3.70 (t, J = 5.3 Hz, 2H), 2.73 (t, J = 4.9 Hz, 2H), 2.48 (s, 3H), 1.49 (s, 9H); MS (ESI) 428 [M+ H] + .
步骤6. 6-甲基-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 6. 6-Methyl-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxylic acid ester:
Figure PCTCN2016087368-appb-000049
Figure PCTCN2016087368-appb-000049
按照实施例1中步骤5的操作方法，以2-(4-(甲氧基羰基)-6-甲基-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(96mg，0.23mmol)为原料去保护基得到淡棕色粉末59mg，产率为79％(二氯甲烷：甲醇＝50：1)。Following the procedure of Step 5 in Example 1, 2-(4-(methoxycarbonyl)-6-methyl-1H-benzo[d]imidazol-2-yl)-4,7-dihydrothiophene And [2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (96 mg, 0.23 mmol) was used as the starting material to give the title compound as a pale brown powder (yield: 79 mg, yield: 79%) :1).
1H NMR(300MHz,Chloroform-d)δ10.34(s,1H),7.74(dd,J＝1.5,0.8Hz,1H),7.69(dd,J＝1.5,0.7Hz,1H),7.36(s,1H),4.09(s,2H),4.00(s,3H),3.16(t,J＝5.8Hz,2H),2.70(t,J＝5.9Hz,2H),2.50(s,3H)；MS(ESI)328[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ10.34 (s, 1H), 7.74 (dd, J = 1.5,0.8Hz, 1H), 7.69 (dd, J = 1.5,0.7Hz, 1H), 7.36 (s , 1H), 4.09 (s, 2H), 4.00 (s, 3H), 3.16 (t, J = 5.8 Hz, 2H), 2.70 (t, J = 5.9 Hz, 2H), 2.50 (s, 3H); (ESI) 328 [M+H] + .
步骤7. 6-甲基-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-7)：Step 7. 6-Methyl-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide (I-7):
Figure PCTCN2016087368-appb-000050
Figure PCTCN2016087368-appb-000050
按照制备化合物I-1的操作方法，以6-甲基-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-羧酸甲酯(59mg，0.15mmol)为原料氨解得到棕色固体48mg，产率为67％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 6-methyl-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[ d] Methyl imidazole-4-carboxylate (59 mg, 0.15 mmol) was obtained as a crude material to give a brown solid (yield: 48%) (yield: methylene chloride:methanol = 10:1).
1H NMR(500MHz,DMSO-d6)δ9.09(brs,1H),7.79(s,1H),7.75(brs,1H),7.64(s,1H),7.49(s,1H),4.22(s,2H),3.22(t,J＝5.8Hz,2H),2.84((t,J＝5.8Hz,2H),2.44(s,3H)；13C NMR(125MHz,DMSO-d6)δ166.75,147.45,139.87,135.96,134.71,133.37,132.29,130.91,128.18,124.58,121.66,115.19,42.77,41.46,23.69,21.67；HRMS(ESI): m/z Calcd.For C16H17N4OS[M+H]+:313.1123；Found:313.1119。 1 H NMR (500MHz, DMSO- d 6) δ9.09 (brs, 1H), 7.79 (s, 1H), 7.75 (brs, 1H), 7.64 (s, 1H), 7.49 (s, 1H), 4.22 ( s, 2H), 3.22 (t, J = 5.8 Hz, 2H), 2.84 ((t, J = 5.8 Hz, 2H), 2.44 (s, 3H); 13 C NMR (125 MHz, DMSO-d 6 ) δ 166. 75,147.45,139.87,135.96,134.71,133.37,132.29,130.91,128.18,124.58,121.66,115.19,42.77,41.46,23.69,21.67;HRMS(ESI): m/z Calcd.For C 16 H 17 N 4 OS[M +H] + : 313.1123; Found: 313.1119.
实施例8. 6-氟-2-(4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-8的制备：Example 8. 6-Fluoro-2-(4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide , Preparation of I-8:
步骤1. 5-氟-7-硝基二氢吲哚-2,3-二酮：Step 1. 5-Fluoro-7-nitroindoline-2,3-dione:
Figure PCTCN2016087368-appb-000051
Figure PCTCN2016087368-appb-000051
按照实施例6中步骤1的操作方法，以5-氟靛红(10.0g，60.6mmol)为原料，硝化制得黄色固体10.6g，产率为83％。According to the procedure of Step 1 in Example 6, 5-fluoroindigo (10.0 g, 60.6 mmol) was used as a raw material to obtain 10.4 g of a yellow solid.
1H NMR(300MHz,DMSO-d6)δ11.71(s,1H),8.21(dd,J＝9.1,2.7Hz,1H),7.97(dd,J＝6.4,2.7Hz,1H)；MS(ESI)211[M+H]+。 1 H NMR (300 MHz, DMSO-d 6 ) δ 11.71 (s, 1H), 8.21. (dd, J = 9.1, 2.7 Hz, 1H), 7.97 (dd, J = 6.4, 2.7 Hz, 1H); ESI) 211 [M+H] + .
步骤2. 2-氨基-5-氟-3-硝基苯甲酸：Step 2. 2-Amino-5-fluoro-3-nitrobenzoic acid:
Figure PCTCN2016087368-appb-000052
Figure PCTCN2016087368-appb-000052
按照实施例6中步骤2的操作方法，以5-氟-7-硝基二氢吲哚-2,3-二酮(10.6g，50.0mmol)为原料制得黄色固体7.0g，产率为70％。According to the procedure of Step 2 in Example 6, a yellow solid (7.0 g) was obtained from 5-fluoro-7-nitroindoline-2,3-dione (10.6 g, 50.0 mmol). 70%.
1H NMR(300MHz,DMSO-d6)δ8.33(brs,2H),8.17(dd,J＝8.8,3.3Hz,1H),8.04(dd,J＝8.6,3.3Hz,1H)；MS(ESI)201[M+H]+。 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.33 (brs, 2H), 8.17 (dd, J = 8.8, 3.3 Hz, 1H), 8.04 (dd, J = 8.6, 3.3 Hz, 1H); ESI) 201 [M+H] + .
步骤3. 2-氨基-5-氟-3-硝基苯甲酸甲酯：Step 3. Methyl 2-amino-5-fluoro-3-nitrobenzoate:
Figure PCTCN2016087368-appb-000053
Figure PCTCN2016087368-appb-000053
按照实施例6中步骤3的操作方法，以2-氨基-5-氟-3-硝基苯甲酸(6.0g，30.0mmol)为原料制得黄色固体2.5g，产率为39％(石油醚：乙酸乙酯＝4：1)。According to the procedure of Step 3 in Example 6, 2-amino-5-fluoro-3-nitrobenzoic acid (6.0 g, 30.0 mmol) was used as the starting material to obtain 2.5 g of a yellow solid, yield 39% ( petroleum ether : ethyl acetate = 4:1).
1H NMR(300MHz,Chloroform-d)δ8.31(brs,2H),8.14(dd,J＝8.4,3.2Hz,1H),8.02(dd,J＝8.3,3.2Hz,1H)；MS(ESI)215[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ8.31 (brs, 2H), 8.14 (dd, J = 8.4,3.2Hz, 1H), 8.02 (dd, J = 8.3,3.2Hz, 1H); MS (ESI ) 215 [M+H] + .
步骤3. 2,3-二氨基-5-氟苯甲酸甲酯：Step 3. Methyl 2,3-diamino-5-fluorobenzoate:
Figure PCTCN2016087368-appb-000054
Figure PCTCN2016087368-appb-000054
按照实施例6中步骤4的操作方法，以2-氨基-5-氟-3-硝基苯甲酸甲酯(2.5g，11.7mmol)为原料制得灰白色固体1.5g，产率为70％(石油醚：乙酸乙酯＝4：1)。1.5 g of an off-white solid was obtained from the methyl 2-amino-5-fluoro-3-nitrobenzoate (2.5 g, 11.7 mmol). Petroleum ether: ethyl acetate = 4:1).
1H NMR(300MHz,Chloroform-d)δ7.11(dd,J＝9.8,2.9Hz,1H),6.62(dd,J＝9.1, 2.9Hz,1H),5.32(brs,2H),3.87(s,3H),3.51(brs,2H)；MS(ESI):185[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 7.11 (dd, J = 9.8, 2.9 Hz, 1H), 6.62 (dd, J = 9.1, 2.9 Hz, 1H), 5.32 (brs, 2H), 3.87 (s) , 3H), 3.51 (brs, 2H); MS (ESI): 185 [M+H] + .
步骤4. 2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯：Step 4. 2-(6-Fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-6,7-dihydrothieno[3,2-c]pyridine- 5(4H)-tert-butyl carboxylic acid:
Figure PCTCN2016087368-appb-000055
Figure PCTCN2016087368-appb-000055
按照实施例1中步骤4的操作方法，以2-甲酰基-6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯(726mg，2.72mmol)和2,3-二氨基-5-氟苯甲酸甲酯(500mg，2.72mmol)为原料制备得到白色粉末1.06g，产率为90％(石油醚：乙酸乙酯＝3：1)。Following the procedure of Step 4 in Example 1, 2-formyl-6,7-dihydrothieno[3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl ester (726 mg, 2.72 mmol) A white powder of 1.06 g (yield: petroleum ether: ethyl acetate = 3:1) was obtained from methylene chloride (2,3-diamino-5-fluorobenzoic acid) (500 mg, 2.72 mmol).
1H NMR(400MHz,Chloroform-d)δ10.40(brs,1H),7.63(dd,J＝8.8,2.4Hz,1H),7.59(dd,J＝9.6,2.4Hz,1H),7.38(s,1H),4.55(s,2H),4.02(s,3H),3.77(t,J＝5.1Hz,2H),2.92(t,J＝5.2Hz,2H),1.50(s,9H)；MS(ESI):432[M+H]+。 1 H NMR (400 MHz, Chloroform-d) δ 10.40 (brs, 1H), 7.63 (dd, J = 8.8, 2.4 Hz, 1H), 7.59 (dd, J = 9.6, 2.4 Hz, 1H), 7.38 (s) , 1H), 4.55 (s, 2H), 4.02 (s, 3H), 3.77 (t, J = 5.1 Hz, 2H), 2.92 (t, J = 5.2 Hz, 2H), 1.50 (s, 9H); (ESI): 432 [M+H] + .
步骤5. 6-氟-2-(4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 5. 6-Fluoro-2-(4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxylic acid methyl ester :
Figure PCTCN2016087368-appb-000056
Figure PCTCN2016087368-appb-000056
按照实施例1中步骤5的操作方法，以2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯(1.0g，2.3mmol)为原料去保护基得到黄色泡沫状固体670mg，产率为88％(二氯甲烷：甲醇＝50：1)。Following the procedure of Step 5 in Example 1, 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-6,7-dihydrothiophene [3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl ester (1.0 g, 2.3 mmol) was used as a starting material to afford the title compound to afford y. 50:1).
1H NMR(300MHz,DMSO-d6)δ7.97(s,1H),7.74(d,J＝9.2Hz,1H),7.54(d,J＝9.2Hz,1H),3.99(s,3H),3.82(s,2H),3.00(t,J＝4.8Hz,2H),2.76(t,J＝4.8Hz,2H)；MS(ESI):332[M+H]+。 1 H NMR (300MHz, DMSO- d 6) δ7.97 (s, 1H), 7.74 (d, J = 9.2Hz, 1H), 7.54 (d, J = 9.2Hz, 1H), 3.99 (s, 3H) , 3.82 (s, 2H), 3.00 (t, J = 4.8 Hz, 2H), 2.76 (t, J = 4.8 Hz, 2H); MS (ESI): 332 [M+H] + .
步骤6. 6-氟-2-(4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-8)：Step 6. 6-Fluoro-2-(4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide ( I-8):
Figure PCTCN2016087368-appb-000057
Figure PCTCN2016087368-appb-000057
按照制备化合物I-1的操作方法，以6-氟-2-(4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(100mg，0.30mmol)为原料氨解得到棕色固体78mg，产率为82％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 6-fluoro-2-(4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d Ammonium imidazole-4-carboxylate (100 mg, 0.30 mmol) was obtained as a crude material to give a brown solid (yield: 78%) (yield: methylene chloride:methanol = 10:1).
1H NMR(500MHz,DMSO-d6)δ9.02(brs,1H),7.88(brs,1H),7.61(s,1H),7.52(dd,J＝10.0,2.5Hz,1H),7.50(dd,J＝8.0,2.5Hz,1H),3.79(s,2H),2.99(t,J＝5.2Hz, 2H),2.74(t,J＝5.2Hz,2H)；13C NMR(125MHz,DMSO-d6)δ165.69,158.58(d,J＝236.0Hz,C-F),149.91,138.63,137.79,136.69,128.92,126.80,122.55,110.05(d,J＝26.5Hz,CH-CF),102.57(d,J＝25.2Hz,CH-CF),79.63,45.19,43.30,25.93；HRMS(ESI):m/z Calcd.For C15H14FN4OS[M+H]+:317.0872；Found:317.0867。 1 H NMR (500 MHz, DMSO-d 6 ) δ 9.02 (brs, 1H), 7.78 (brs, 1H), 7.61 (s, 1H), 7.52 (dd, J = 10.0, 2.5 Hz, 1H), 7.50 ( Dd, J = 8.0, 2.5 Hz, 1H), 3.79 (s, 2H), 2.99 (t, J = 5.2 Hz, 2H), 2.74 (t, J = 5.2 Hz, 2H); 13 C NMR (125 MHz, DMSO) -d 6 ) δ 165.69, 158.58 (d, J = 236.0 Hz, CF), 149.91, 138.63, 137.79, 136.69, 128.92, 126.80, 122.55, 110.05 (d, J = 26.5 Hz, CH-CF), 102.57 (d, J = 25.2 Hz, CH-CF), 79.63, 45.19, 43.30, 25.93; HRMS (ESI): m/z Calcd. For C 15 H 14 FN 4 OS [M+H] + : 317.0872; Found: 317.0867.
实施例9. 6-氟-2-(5-甲基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-9的制备：Example 9. 6-Fluoro-2-(5-methyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole Preparation of 4-carboxamide, I-9:
步骤1. 6-氟-2-(5-甲基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 1. 6-Fluoro-2-(5-methyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole- 4-methyl formate:
Figure PCTCN2016087368-appb-000058
Figure PCTCN2016087368-appb-000058
按照实施例2中步骤1的操作方法，以6-氟-2-(4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(198mg，0.60mmol)为原料，经甲基化得黄色固体200mg，产率为97％(二氯甲烷：甲醇＝30：1)。Following the procedure of Step 1 in Example 2, 6-fluoro-2-(4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[ d] Methyl imidazole-4-carboxylate (198 mg, 0.60 mmol) was obtained as a material.
1H NMR(400MHz,Chloroform-d)δ7.63(dd,J＝8.9,2.4Hz,1H),7.58(dd,J＝9.7,2.4Hz,1H),7.34(s,1H),4.02(s,3H),3.55(s,2H),2.99(t,J＝5.7Hz,2H),2.79(t,J＝5.7Hz,2H),2.51(s,3H)；MS(ESI)346[M+H]+。 1 H NMR (400 MHz, Chloroform-d) δ 7.63 (dd, J = 8.9, 2.4 Hz, 1H), 7.58 (dd, J = 9.7, 2.4 Hz, 1H), 7.34 (s, 1H), 4.02 (s) , 3H), 3.55 (s, 2H), 2.99 (t, J = 5.7 Hz, 2H), 2.79 (t, J = 5.7 Hz, 2H), 2.51 (s, 3H); MS (ESI) 346 [M+ H] + .
步骤2. 6-氟-2-(5-甲基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-9)：Step 2. 6-Fluoro-2-(5-methyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole- 4-formamide (I-9):
Figure PCTCN2016087368-appb-000059
Figure PCTCN2016087368-appb-000059
按照制备化合物I-1的操作方法，以6-氟-2-(5-甲基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(100mg，0.29mmol)为原料氨解得到浅黄色粉末58mg，产率为61％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 6-fluoro-2-(5-methyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H Methyl benzo[d]imidazole-4-carboxylate (100 mg, 0.29 mmol) was obtained as a crude material (yield: 58 mg,yield: 61% (dichloromethane:methanol = 10:1).
1H NMR(400MHz,DMSO-d6)δ9.08(brs,1H),7.95(s,1H),7.65(s,1H),7.58–7.50(m,2H),3.57(s,1H),2.92(t,J＝5.1Hz,2H),2.79(t,J＝5.0Hz,2H),2.45(s,3H)；13C NMR(125MHz,DMSO-d6)δ165.35,158.78(d,J＝237.0Hz,C-F),149.18,138.48,137.68,136.04,135.24,129.62,126.82,123.20,110.38(d,J＝24.0Hz,CH-CF),101.78(d,J＝25.0Hz,CH-CF),54.18,52.00,45.11,25.31；HRMS(ESI):m/z Calcd.For C16H16FN4OS[M+H]+:331.1029；Found:331.1022。 1 H NMR (400MHz, DMSO- d 6) δ9.08 (brs, 1H), 7.95 (s, 1H), 7.65 (s, 1H), 7.58-7.50 (m, 2H), 3.57 (s, 1H), 2.92 (t, J = 5.1 Hz, 2H), 2.79 (t, J = 5.0 Hz, 2H), 2.45 (s, 3H); 13 C NMR (125 MHz, DMSO-d 6 ) δ 165.35, 158.78 (d, J = 237.0 Hz, CF), 149.18, 138.48, 137.68, 136.04, 135.24, 129.62, 126.82, 123.20, 110.38 (d, J = 24.0 Hz, CH-CF), 101.78 (d, J = 25.0 Hz, CH-CF), 54.18, 52.00, 45.11, 25.31; HRMS (ESI): m/z Calcd. For C 16 H 16 FN 4 OS [M+H] + : 331.1029; Found: 331.022.
实施例10. 2-(5-乙酰基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑 -4-甲酰胺，I-10的制备：Example 10. 2-(5-Acetyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-6-fluoro-1H-benzo[d]imidazole Preparation of 4-carboxamide, I-10:
步骤1. 2-(5-乙酰基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯：Step 1. 2-(5-Acetyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-6-fluoro-1H-benzo[d]imidazole- 4-methyl formate:
Figure PCTCN2016087368-appb-000060
Figure PCTCN2016087368-appb-000060
0℃下，向溶有6-氟-2-(4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(198mg，0.60mmol)与三乙胺(121mg，1.20mmol)的无水DMF(4mL)溶液中缓慢滴加乙酰氯(57mg，0.72mmol)，30分钟后，将反应液倒入冰水混合物中，析出白色固体，过滤，真空干燥得197mg，产率为88％。6-fluoro-2-(4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d]imidazole-4 was dissolved at 0 °C - acetyl chloride (57 mg, 0.72 mmol) was slowly added dropwise to a solution of methyl formate (198 mg, 0.60 mmol) and triethylamine (121 mg, 1.20 mmol) in anhydrous DMF (4 mL). In a mixture of ice and water, a white solid was precipitated, filtered, and dried in vacuo to give 197 mg.
1H NMR(400MHz,Chloroform-d)构象异构体1：δ10.50(brs,1H),7.65–7.57(m,2H),7.40(s,1H),4.71(s,2H),4.02(s,3H),3.79(t,J＝5.7Hz,2H),2.99(t,J＝5.6Hz,2H),2.22(s,3H)；构象异构体2：δ10.43(brs,1H),7.65–7.57(m,2H),7.40(s,1H),4.59(s,2H),4.02(s,3H),3.96(t,J＝5.7Hz,2H),2.93(t,J＝5.6Hz,2H),2.20(s,3H)；MS(ESI)374[M+H]+。 1 H NMR (400MHz, Chloroform- d) conformational isomers 1: δ10.50 (brs, 1H) , 7.65-7.57 (m, 2H), 7.40 (s, 1H), 4.71 (s, 2H), 4.02 ( s, 3H), 3.79 (t, J = 5.7 Hz, 2H), 2.99 (t, J = 5.6 Hz, 2H), 2.22 (s, 3H); conformation 2: δ 10.43 (brs, 1H) , 7.65–7.57 (m, 2H), 7.40 (s, 1H), 4.59 (s, 2H), 4.02 (s, 3H), 3.96 (t, J = 5.7 Hz, 2H), 2.93 (t, J = 5.6 Hz, 2H), 2.20 (s, 3H); MS (ESI) 374 [M+H] + .
步骤2. 2-(5-乙酰基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺(I-10)Step 2. 2-(5-Acetyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-6-fluoro-1H-benzo[d]imidazole- 4-carboxamide (I-10)
Figure PCTCN2016087368-appb-000061
Figure PCTCN2016087368-appb-000061
按照制备化合物I-1的操作方法，以2-(5-乙酰基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯(197mg，0.53mmol)为原料氨解得到白色粉末182mg，产率为96％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 2-(5-acetyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-6-fluoro-1H - Benzo[d]imidazole-4-carboxylic acid methyl ester (197 mg, 0.53 mmol) was obtained as a crude material (yield: 182 mg,yield: 96% (dichloromethane:methanol = 10:1).
1H NMR(300MHz,DMSO-d6)构象异构体1：δ9.05(brs,1H),7.92(s,1H),7.71(brs,1H),7.60–7.50(m,2H),4.63(s,2H),3.75(m,2H),2.82(brs,2H),2.10(s,3H)；构象异构体2：δ9.05(brs,1H),7.92(s,1H),7.71(brs,1H),7.60–7.50(m,2H),4.58(s,2H),7.82–7.71(m,2H),2.95(brs,2H),2.12(s,3H)；13C NMR(125MHz,DMSO-d6)构象异构体1：δ169.44,165.52,158.68(d,J＝229.0Hz,C-F),149.23,138.18,137.75,134.39,134.06,130.01,126.65(d,J＝25.2Hz,CH-CF),110.38(d,J＝25.2Hz,CH-CF),45.90,43.75,25.04,22.31；构象异构体2：δ169.38,165.52,158.68(d,J＝229.0Hz,C-F),149.23,138.18,137.75,134.39,134.06,130.01,126.65(d,J＝25.2Hz,CH-CF),110.38(d,J＝25.2Hz,CH-CF),45.90,41.93,25.77,21.87；HRMS(ESI):m/z Calcd.For C17H16FN4O2S [M+H]+:359.0978；Found:359.0973。 1 H NMR (300 MHz, DMSO-d 6 ). </ RTI></RTI></RTI> δ 9.05 (brs, 1H), 7.92 (s, 1H), 7.71 (brs, 1H), 7.60 - 7.50 (m, 2H), 4.63 (s, 2H), 3.75 (m, 2H), 2.82 (brs, 2H), 2.10 (s, 3H); conformation 2: δ 9.05 (brs, 1H), 7.92 (s, 1H), 7.71 (brs, 1H), 7.60–7.50 (m, 2H), 4.58 (s, 2H), 7.82–7.71 (m, 2H), 2.95 (brs, 2H), 2.12 (s, 3H); 13 C NMR (125 MHz) , DMSO-d 6 ) conformational isomer 1: δ 169.44, 165.52, 158.68 (d, J = 229.0 Hz, CF), 149.23, 138.18, 137.75, 134.39, 134.06, 130.01, 126.65 (d, J = 25.2 Hz, CH -CF), 110.38 (d, J = 25.2 Hz, CH-CF), 45.90, 43.75, 25.04, 22.31; conformational isomer 2: δ 169.38, 165.52, 158.68 (d, J = 229.0 Hz, CF), 149.23, 138.18, 137.75, 134.39, 134.06, 130.01, 126.65 (d, J = 25.2 Hz, CH-CF), 110.38 (d, J = 25.2 Hz, CH-CF), 45.90, 41.93, 25.77, 21.87; HRMS (ESI) : m/z Calcd. For C 17 H 16 FN 4 O 2 S [M+H] + : 359.0978; Found: 359.0973.
实施例11. 2-(5-(环丙基羰基)-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺，I-11的制备：Example 11. 2-(5-(Cyclopropylcarbonyl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-6-fluoro-1H-benzo [d] Preparation of imidazole-4-carboxamide, I-11:
步骤1. 2-(5-(环丙基羰基)-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯：Step 1. 2-(5-(Cyclopropylcarbonyl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-6-fluoro-1H-benzo[ d] Methyl imidazole-4-carboxylate:
Figure PCTCN2016087368-appb-000062
Figure PCTCN2016087368-appb-000062
按照实施例10中步骤1的操作方法，以6-氟-2-(4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(182mg，0.60mmol)和环丙基甲酰氯(75mg，0.72mmol)为原料制得白色粉末182mg，产率为76％。Following the procedure of Step 1 in Example 10, 6-fluoro-2-(4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[ d] Methyl imidazole-4-carboxylate (182 mg, 0.60 mmol) and cyclopropylcarbonyl chloride (75 mg, 0.72 mmol) were obtained as a white powder (yield: 182 mg).
1H NMR(300MHz,CD3OD)构象异构体1：δ7.89(s,1H),7.73(s,1H),7.59(dd,J＝9.8,2.3Hz,1H),7.52(dd,J＝8.5,2.3Hz,1H),4.92(s,2H),4.02(s,3H),3.93(t,J＝5.9Hz,2H),2.90(t,J＝5.7Hz,2H),2.10(m,1H),0.96–0.85(m,4H)；构象异构体2：δ7.89(s,1H),7.73(s,1H),7.59(dd,J＝9.8,2.3Hz,1H),7.52(dd,J＝8.5,2.3Hz,1H),4.68(s,2H),4.09(t,J＝5.9Hz,2H),4.02(s,3H),3.04(t,J＝5.8Hz,2H),2.10(m,1H),0.96–0.85(m,4H)；MS(ESI)400[M+H]+。 1 H NMR (300 MHz, CD 3 OD) conformer 1 : δ 7.89 (s, 1H), 7.73 (s, 1H), 7.59 (dd, J = 9.8, 2.3 Hz, 1H), 7.52 (dd, J = 8.5, 2.3 Hz, 1H), 4.92 (s, 2H), 4.02 (s, 3H), 3.93 (t, J = 5.9 Hz, 2H), 2.90 (t, J = 5.7 Hz, 2H), 2.10 ( m,1H), 0.96–0.85 (m, 4H); conformational isomer 2: δ 7.89 (s, 1H), 7.73 (s, 1H), 7.59 (dd, J = 9.8, 2.3 Hz, 1H), 7.52 (dd, J = 8.5, 2.3 Hz, 1H), 4.68 (s, 2H), 4.09 (t, J = 5.9 Hz, 2H), 4.02 (s, 3H), 3.04 (t, J = 5.8 Hz, 2H) ), 2.10 (m, 1H), 0.96 - 0.85 (m, 4H); MS (ESI) 400 [M+H] + .
步骤2. 2-(5-(环丙基羰基)-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺(I-11)：Step 2. 2-(5-(Cyclopropylcarbonyl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-6-fluoro-1H-benzo[ d] Imidazole-4-carboxamide (I-11):
Figure PCTCN2016087368-appb-000063
Figure PCTCN2016087368-appb-000063
按照制备化合物I-1的操作方法，以2-(5-(环丙基羰基)-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯(182mg，0.46mmol)为原料氨解得到浅棕色固体106mg，产率为60％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 2-(5-(cyclopropylcarbonyl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-6 -Fluoro-1H-benzo[d]imidazole-4-carboxylic acid methyl ester (182 mg, 0.46 mmol) was obtained as a crude material to give a pale brown solid (yield: 60%) (yield: methylene chloride:methanol = 10:1).
1H NMR(400MHz,DMSO-d6)构象异构体1：δ9.06(brs,1H),7.94(s,1H),7.72(brs,1H),7.60–7.52(m,2H),4.88(s,2H),3.82(brs,2H),2.83(brs,2H),2.10(m,1H),0.75(m,4H)；构象异构体2：δ9.06(brs,1H),7.94(s,1H),7.72(brs,1H),7.60–7.52(m,2H),4.60(s,2H),4.01(brs,2H),2.98(brs,2H),2.10(m,1H),0.77(m,4H)；13C NMR(125MHz,DMSO-d6)构象异构体1：δ172.17,165.39,158.74(d,J＝236.9Hz,C-F),149.18,138.31,137.84,136.01,134.55,134.17,130.10,126.66(d,J＝25.2Hz,CH-CF),123.41,110.54(d,J＝26.0Hz,CH-CF),102.01,45.21,43.07,26.17,11.34,7.86；构象异构 体2：δ172.17,165.39,158.74(d,J＝236.9Hz,C-F),149.18,138.31,137.84,136.01,134.55,134.17,130.10,126.66(d,J＝25.2Hz,CH-CF),123.41,110.54(d,J＝26.0Hz,CH-CF),102.01,49.05,42.75,25.01,11.06,7.49；HRMS(ESI):m/z Calcd.For C19H18FN4O2S[M+H]+:385.1134；Found:385.1125。 1 H NMR (400MHz, DMSO- d 6) conformational isomers 1: δ9.06 (brs, 1H) , 7.94 (s, 1H), 7.72 (brs, 1H), 7.60-7.52 (m, 2H), 4.88 (s, 2H), 3.82 (brs, 2H), 2.83 (brs, 2H), 2.10 (m, 1H), 0.75 (m, 4H); conformation 2: δ 9.06 (brs, 1H), 7.94 (s, 1H), 7.72 (brs, 1H), 7.60 - 7.52 (m, 2H), 4.60 (s, 2H), 4.01 (brs, 2H), 2.98 (brs, 2H), 2.10 (m, 1H), 0.77 (m, 4H); 13 C NMR (125MHz, DMSO-d 6 ). </ RTI></RTI></RTI> δ 172.17, 165.39, 158.74 (d, J = 236.9 Hz, CF), 149.18, 138.31, 137.84, 136.01, 134.55, 134.17, 130.10, 126.66 (d, J = 25.2 Hz, CH-CF), 123.41, 110.54 (d, J = 26.0 Hz, CH-CF), 102.01, 45.21, 43.07, 26.17, 11.34, 7.86; conformational isomer 2: δ 172.17, 165.39, 158.74 (d, J = 236.9 Hz, CF), 149.18, 138.31, 137.84, 136.01, 134.55, 134.17, 130.10, 126.66 (d, J = 25.2 Hz, CH-CF), 123.41, 110.54 ( d, J = 26.0 Hz, CH-CF), 102.01, 49.05, 42.75, 25.01, 11.06, 7.49; HRMS (ESI): m/z Calcd. For C 19 H 18 FN 4 O 2 S[M+H] + :385.1134;Found:385.1125.
实施例12. 6-氟-2-(5-(2-丙氧基乙酰基)-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-12的制备：Example 12. 6-Fluoro-2-(5-(2-propoxyacetyl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H -Benzo[d]imidazole-4-carboxamide, preparation of I-12:
步骤1. 6-氟-2-(5-(2-丙氧基乙酰基)-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 1. 6-Fluoro-2-(5-(2-propoxyacetyl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H- Benzo[d]imidazole-4-carboxylic acid methyl ester:
Figure PCTCN2016087368-appb-000064
Figure PCTCN2016087368-appb-000064
按照实施例10中步骤1的操作方法，以6-氟-2-(4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(182mg，0.60mmol)和2-丙氧基乙酰氯(98mg，0.72mmol)为原料制得白色粉末228mg，产率为88％。Following the procedure of Step 1 in Example 10, 6-fluoro-2-(4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[ d] Methyl imidazole-4-carboxylate (182 mg, 0.60 mmol) and 2-propoxyacetyl chloride (98 mg, 0.72 mmol) were obtained as a white powder (yield: 228 mg).
1H NMR(300MHz,DMSO-d6)δ8.09(s,1H),7.77(d,J＝8.7Hz,1H),7.56(d,J＝9.3Hz,1H),4.60(s,2H),4.23(s,2H),3.99(s,3H),3.78(t,J＝5.1Hz,2H),3.41(t,J＝5.1Hz,2H),2.90(brs,2H),1.60–1.46(m,2H),0.92–0.82(m,3H)；MS(ESI)432[M+H]+。 1 H NMR (300MHz, DMSO- d 6) δ8.09 (s, 1H), 7.77 (d, J = 8.7Hz, 1H), 7.56 (d, J = 9.3Hz, 1H), 4.60 (s, 2H) , 4.23 (s, 2H), 3.99 (s, 3H), 3.78 (t, J = 5.1 Hz, 2H), 3.41 (t, J = 5.1 Hz, 2H), 2.90 (brs, 2H), 1.60 - 1.46 ( m, 2H), 0.92 - 0.82 (m, 3H); MS (ESI) 432 [M+H] + .
步骤2. 6-氟-2-(5-(2-丙氧基乙酰基)-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-12)：Step 2. 6-Fluoro-2-(5-(2-propoxyacetyl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H- Benzo[d]imidazole-4-carboxamide (I-12):
Figure PCTCN2016087368-appb-000065
Figure PCTCN2016087368-appb-000065
按照制备化合物I-1的操作方法，以6-氟-2-(5-(2-丙氧基乙酰基)-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(228mg，0.53mmol)为原料氨解得到浅棕色固体207mg，产率为94％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 6-fluoro-2-(5-(2-propoxyacetyl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridine The methyl group of 2-yl)-1H-benzo[d]imidazole-4-carboxylate (228 mg, 0.53 mmol) was obtained as a crude material (yield: 207 mg,yield: 94%, m. 1).
1H NMR(500MHz,DMSO-d6)δ8.96(brs,1H),7.90(s,1H),7.77–7.43(m,3H),4.57(s,2H),4.21(s,2H),3.85–3.69(m,2H),3.39(s,2H),3.05–2.75(m,2H),1.51(q,J＝6.2Hz,2H),0.85(t,J＝6.5Hz,3H)；13C NMR(125MHz,DMSO-d6)构象异构体1：δ168.55,165.64,158.67(d,J＝236.9Hz,C-F),149.38,138.57,137.54,136.29,133.99,130.21,126.70(d,J＝27.7Hz,CH-CF),110.29(d,J＝25.2Hz,CH-CF),102.68,72.65,70.10,44.51,42.49,25.85,22.85,10.99；构象异构体2：δ8.96(brs,1H),7.90(s,1H), 7.77–7.43(m,3H),4.57(s,2H),4.21(s,2H),3.85–3.69(m,2H),3.39(s,2H),3.05–2.75(m,2H),1.51(q,J＝6.2Hz,2H),0.85(t,J＝6.5Hz,3H)；HRMS(ESI):m/z Calcd.For C20H22FN4O3S[M+H]+:417.1397；Found:417.1398。 1 H NMR (500MHz, DMSO- d 6) δ8.96 (brs, 1H), 7.90 (s, 1H), 7.77-7.43 (m, 3H), 4.57 (s, 2H), 4.21 (s, 2H), 3.85-3.69 (m, 2H), 3.39 (s, 2H), 3.05-2.75 (m, 2H), 1.51 (q, J = 6.2Hz, 2H), 0.85 (t, J = 6.5Hz, 3H); 13 C NMR (125 MHz, DMSO-d 6 ). </ RTI></RTI> δ 168.55, 165.64, 158.67 (d, J = 236.9 Hz, CF), 149.38, 138.57, 137.54, 136.29, 133.99, 130.21, 126.70 (d, J = 27.7 Hz, CH-CF), 110.29 (d, J = 25.2 Hz, CH-CF), 102.68, 72.65, 70.10, 44.51, 42.49, 25.85, 22.85, 10.99; conformational isomer 2: δ 8.96 (brs, 1H), 7.90 (s, 1H), 7.77 - 7.43 (m, 3H), 4.57 (s, 2H), 4.21 (s, 2H), 3.85 - 3.69 (m, 2H), 3.39 (s, 2H), 3.05 -2.75 (m, 2H), 1.51 (q, J = 6.2 Hz, 2H), 0.85 (t, J = 6.5 Hz, 3H); HRMS (ESI): m/z Calcd. For C 20 H 22 FN 4 O 3 S[M+H] + : 417.1397; Found: 417.1398.
实施例13. 2-(5-苯甲酰基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺，I-13的制备：Example 13. 2-(5-Benzoyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-6-fluoro-1H-benzo[d] Preparation of imidazole-4-carboxamide, I-13:
步骤1. 2-(5-苯甲酰基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯：Step 1. 2-(5-Benzoyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-6-fluoro-1H-benzo[d]imidazole -4-methyl formate:
Figure PCTCN2016087368-appb-000066
Figure PCTCN2016087368-appb-000066
按照实施例10中步骤1的操作方法，以6-氟-2-(4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(182mg，0.60mmol)和苯甲酰氯(101mg，0.72mmol)为原料制得白色粉末238mg，产率为91％。Following the procedure of Step 1 in Example 10, 6-fluoro-2-(4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[ d] Imidazole-4-carboxylic acid methyl ester (182 mg, 0.60 mmol) and benzoyl chloride (101 mg, 0.72 mmol) were used as starting materials to yield 238 mg of white powder, yield 91%.
1H NMR(400MHz,Chloroform-d)构象异构体1：δ10.5(brs,1H),8.12(d,J＝8.1Hz,1H),7.69–7.57(m,2H),7.50–7.43(m,5H),4.87(s,2H),4.01(s,3H),3.75(brs,2H),2.97(brs,2H)；构象异构体2：δ10.5(brs,1H),8.12(d,J＝8.1Hz,1H),7.69–7.57(m,2H),7.50–7.43(m,5H),4.57(s,2H),4.12(brs,2H),4.01(s,3H),3.06(brs,2H)；MS(ESI)436[M+H]+。 1 H NMR (400 MHz, Chloroform-d) conformation 1 : δ 10.5 (brs, 1H), 8.12 (d, J = 8.1 Hz, 1H), 7.69 - 7.57 (m, 2H), 7.50 - 7.43 ( m,5H), 4.87 (s, 2H), 4.01 (s, 3H), 3.75 (brs, 2H), 2.97 (brs, 2H); conformation 2: δ 10.5 (brs, 1H), 8.12 ( d, J = 8.1 Hz, 1H), 7.69 - 7.57 (m, 2H), 7.50 - 7.43 (m, 5H), 4.57 (s, 2H), 4.12 (brs, 2H), 4.01 (s, 3H), 3.06 (brs, 2H); MS (ESI) 436 [M+H] + .
步骤2. 2-(5-苯甲酰基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺(I-13)：Step 2. 2-(5-Benzoyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-6-fluoro-1H-benzo[d]imidazole 4-carboxamide (I-13):
Figure PCTCN2016087368-appb-000067
Figure PCTCN2016087368-appb-000067
按照制备化合物I-1的操作方法，以2-(5-苯甲酰基-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯(238mg，0.55mmol)为原料氨解得到白色固体159mg，产率为69％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 2-(5-benzoyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-6-fluoro- A solution of 1H-benzo[d]imidazole-4-carboxylic acid methyl ester (238 mg, 0.55 mmol) was obtained as a white solid (yield: 159 mg, yield: 69% (dichloromethane:methanol = 10:1).
1H NMR(400MHz,DMSO-d6)构象异构体1：δ9.07(brs,1H),7.96(s,1H),7.75(brs,1H),7.60–7.52(m,2H),7.52–7.43(m,5H),4.75(s,2H),3.64(brs,2H),2.95(brs,2H)；构象异构体2：δ9.07(brs,1H),7.96(s,1H),7.75(brs,1H),7.60–7.52(m,2H),7.52–7.43(m,5H),4.55(s,2H),3.98(brs,2H),2.95(brs,2H)；13C NMR(125MHz,DMSO-d6)δ170.42,165.32,158.82(d,J＝238.0Hz,C-F),149.01,138.47,137.70,136.46,135.96,133.85,130.24,129.02,127.22,126.75,123.38,110.52(d,J＝23.0Hz,CH-CF), 101.88(d,J＝23.0Hz,CH-CF),45.12,42.69,25.83；HRMS(ESI):m/z Calcd.For C22H18FN4O2S[M+H]+:421.1134；Found:421.1128。 1 H NMR (400MHz, DMSO- d 6) conformational isomers 1: δ9.07 (brs, 1H) , 7.96 (s, 1H), 7.75 (brs, 1H), 7.60-7.52 (m, 2H), 7.52 -7.43 (m, 5H), 4.75 (s, 2H), 3.64 (brs, 2H), 2.95 (brs, 2H); conformation 2: δ 9.07 (brs, 1H), 7.96 (s, 1H) , 7.75 (brs, 1H), 7.60–7.52 (m, 2H), 7.52–7.43 (m, 5H), 4.55 (s, 2H), 3.98 (brs, 2H), 2.95 (brs, 2H); 13 C NMR (125MHz, DMSO-d 6 ) δ 170.42, 165.32, 158.82 (d, J = 238.0 Hz, CF), 149.01, 138.47, 137.70, 136.46, 135.96, 133.85, 130.24, 129.02, 127.22, 126.75, 123.38, 110.52 (d, J = 23.0 Hz, CH-CF), 101.88 (d, J = 23.0 Hz, CH-CF), 45.12, 42.69, 25.83; HRMS (ESI): m/z Calcd. For C 22 H 18 FN 4 O 2 S [M+H] + : 421.1134; Found: 421.1128.
实施例14. 6-氟-2-(5-(甲磺酰基)-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-14的制备：Example 14. 6-Fluoro-2-(5-(methylsulfonyl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[ d] Preparation of imidazole-4-carboxamide, I-14:
步骤1. 6-氟-2-(5-(甲磺酰基)-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 1. 6-Fluoro-2-(5-(methylsulfonyl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d Methyl imidazole-4-carboxylate:
Figure PCTCN2016087368-appb-000068
Figure PCTCN2016087368-appb-000068
按照实施例10中步骤1的操作方法，以6-氟-2-(4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(182mg，0.60mmol)和甲磺酰氯(82mg，0.72mmol)为原料制得白色粉末135mg，产率为55％。Following the procedure of Step 1 in Example 10, 6-fluoro-2-(4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[ d] Methyl imidazole-4-carboxylate (182 mg, 0.60 mmol) and methanesulfonyl chloride (82 mg, 0.72 mmol) were obtained as a white powder, 135 mg, yield 55%.
1H NMR(400MHz,DMSO-d6)δ12.69(brs,1H),8.08(s,1H),7.78(dd,J＝9.0,2.0Hz,1H),7.56(dd,J＝10.0,2.3Hz,1H),4.36(s,2H),3.99(s,3H),3.54(t,J＝5.6Hz,2H),3.00(brs,5H)；MS(ESI)436[M+H]+。 1 H NMR (400MHz, DMSO- d 6) δ12.69 (brs, 1H), 8.08 (s, 1H), 7.78 (dd, J = 9.0,2.0Hz, 1H), 7.56 (dd, J = 10.0,2.3 Hz, 1H), 4.36 (s, 2H), 3.99 (s, 3H), 3.54 (t, J = 5.6 Hz, 2H), 3.00 (brs, 5H); MS (ESI) 436 [M+H] + .
步骤2. 6-氟-2-(5-(甲磺酰基)-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-14)：Step 2. 6-Fluoro-2-(5-(methylsulfonyl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-1H-benzo[d Imidazole-4-carboxamide (I-14):
Figure PCTCN2016087368-appb-000069
Figure PCTCN2016087368-appb-000069
按照制备化合物I-1的操作方法，以6-氟-2-(5-(甲磺酰基)-4,5,6,7-四氢噻吩并[3,2-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(135mg，0.33mmol)为原料氨解得到白色粉末105mg，产率为81％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 6-fluoro-2-(5-(methylsulfonyl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl The methyl ester of -1H-benzo[d]imidazole-4-carboxylate (135 mg, 0.33 mmol) was obtained as a white powder to give a white powder (yield: 81%) (yield: methylene chloride:methanol = 10:1).
1H NMR(300MHz,DMSO-d6)δ8.98(brs,1H),7.92(s,1H),7.73(brs,1H),7.62–7.48(m,2H),4.36(s,2H),3.53(t,J＝5.6Hz,2H),2.99(brs,5H)；13C NMR(125MHz,DMSO-d6)δ165.45,158.73(d,J＝236.0Hz,C-F),149.15,144.29,137.33,133.00,132.99,130.30,126.58,123.27(d,J＝24.2Hz,CH-CF),110.44(d,J＝25.2Hz,CH-CF),102.10,45.39,43.35,35.96,25.37；HRMS(ESI):m/z Calcd.For C16H16FN4O3S2[M]+:395.0648；Found:395.0639。 1 H NMR (300MHz, DMSO- d 6) δ8.98 (brs, 1H), 7.92 (s, 1H), 7.73 (brs, 1H), 7.62-7.48 (m, 2H), 4.36 (s, 2H), 3.53 (t, J = 5.6 Hz, 2H), 2.99 (brs, 5H); 13 C NMR (125MHz, DMSO-d 6 ) δ 165.45, 158.73 (d, J = 236.0 Hz, CF), 149.15, 144.29, 137.33, 133.00, 132.99, 130.30, 126.58, 123.27 (d, J = 24.2 Hz, CH-CF), 110.44 (d, J = 25.2 Hz, CH-CF), 102.10, 45.39, 43.35, 35.96, 25.37; HRMS (ESI) : m/z Calcd. For C 16 H 16 FN 4 O 3 S 2 [M] + : 395.0648; Found: 395.0639.
实施例15. 6-氟-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-15的制备：Example 15. 6-Fluoro-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide Preparation of I-15:
步骤1. 2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶 -6(5H)-羧酸叔丁酯：Step 1. 2-(6-Fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7-dihydrothieno[2,3-c]pyridine -6(5H)-tert-butyl carboxylic acid:
Figure PCTCN2016087368-appb-000070
Figure PCTCN2016087368-appb-000070
按照实施例1中步骤4的操作方法，以2-甲酰基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(726mg，2.72mmol)和2,3-二氨基-5-氟苯甲酸甲酯(500mg，2.72mmol)为原料制备得到黄色泡沫状固体973mg，产率为83％(石油醚：乙酸乙酯＝3：1)。Following the procedure of Step 4 in Example 1, 2-formyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (726 mg, 2.72 mmol) The title compound was obtained as a yellow foamy solid 973 mg (yield: petroleum ether: ethyl acetate = 3:1).
1H NMR(300MHz,DMSO-d6)δ12.60(brs,1H),8.05(s,1H),7.75(dd,J＝9.0,2.1Hz,1H),7.53(dd,J＝9.5,2.1Hz,1H),4.61(s,2H),3.96(s,3H),3.62(t,J＝5.7Hz,2H),2.69(t,J＝5.7Hz,2H),1.42(s,9H)；MS(ESI)432[M+H]+。 1 H NMR (300MHz, DMSO- d 6) δ12.60 (brs, 1H), 8.05 (s, 1H), 7.75 (dd, J = 9.0,2.1Hz, 1H), 7.53 (dd, J = 9.5,2.1 Hz, 1H), 4.61 (s, 2H), 3.96 (s, 3H), 3.62 (t, J = 5.7 Hz, 2H), 2.69 (t, J = 5.7 Hz, 2H), 1.42 (s, 9H); MS (ESI) 432 [M + H] + .
步骤2. 6-氟-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 2. 6-Fluoro-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxylic acid methyl ester :
Figure PCTCN2016087368-appb-000071
Figure PCTCN2016087368-appb-000071
按照实施例1中步骤5的操作方法，以2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(500mg，1.16mmol)为原料去保护基得到浅黄色泡沫状固体303mg，产率为79％(二氯甲烷：甲醇＝50：1)。Following the procedure of Step 5 in Example 1, 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7-dihydrothiophene [2,3-c]pyridin-6(5H)-carboxylic acid tert-butyl ester (500 mg, 1.16 mmol) was obtained as the starting material to give the title compound as a pale yellow foamy solid 303 mg (yield: 50:1).
1H NMR(300MHz,DMSO-d6)δ8.00(brs,1H),7.72(dd,J＝9.4,2.1Hz,1H),7.52(dd,J＝9.9,2.1Hz,1H),3.96(s,3H),3.90(s,2H),2.94(t,J＝5.7Hz,2H),2.59(t,J＝5.6Hz,2H)；MS(ESI)332[M+H]+。 1 H NMR (300MHz, DMSO- d 6) δ8.00 (brs, 1H), 7.72 (dd, J = 9.4,2.1Hz, 1H), 7.52 (dd, J = 9.9,2.1Hz, 1H), 3.96 ( s, 3H), 3.90 (s, 2H), 2.94 (t, J = 5.7 Hz, 2H), 2.59 (t, J = 5.6 Hz, 2H); MS (ESI) 332 [M+H] + .
步骤3. 6-氟-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-15)：Step 3. 6-Fluoro-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide ( I-15):
Figure PCTCN2016087368-appb-000072
Figure PCTCN2016087368-appb-000072
按照制备化合物I-1的操作方法，以6-氟-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(100mg，0.30mmol)为原料氨解得到白色粉末72mg，产率为76％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 6-fluoro-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d] Ammonium imidazole-4-carboxylate (100 mg, 0.30 mmol) was obtained as a crude material to give a white powder (yield: 76%) (yield: methylene chloride:methanol = 10:1).
1H NMR(500MHz,DMSO-d6)δ9.05(brs,1H),7.94(s,1H),7.72(brs,1H),7.61–7.54(m,2H),3.96(s,2H),2.99(brs,2H),2.65(brs,2H)；13C NMR(125MHz,DMSO-d6)δ165.62,158.60(d,J＝234.0Hz,C-F),149.79,139.68,135.98,129.11,128.95 (d,J＝26.0Hz,CH-CF),110.12(d,J＝25.8Hz,CH-CF),44.77,42.92,26.47；HRMS(ESI):m/z Calcd.For C15H14FN4OS[M+H]+:317.0872；Found:317.0861。 1 H NMR (500MHz, DMSO- d 6) δ9.05 (brs, 1H), 7.94 (s, 1H), 7.72 (brs, 1H), 7.61-7.54 (m, 2H), 3.96 (s, 2H), 2.99 (brs, 2H), 2.65 (brs, 2H); 13 C NMR (125MHz, DMSO-d 6 ) δ 165.62, 158.60 (d, J = 234.0 Hz, CF), 149.79, 139.68, 135.98, 129.11, 128.95 (d , J = 26.0 Hz, CH-CF), 110.12 (d, J = 25.8 Hz, CH-CF), 44.77, 42.92, 26.47; HRMS (ESI): m/z Calcd. For C 15 H 14 FN 4 OS [ M+H] + : 317.0872; Found: 317.0861.
实施例16. 6-氟-2-(6-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-16的制备：Example 16. 6-Fluoro-2-(6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole Preparation of 4-carboxamide, I-16:
步骤1. 6-氟-2-(6-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 1. 6-Fluoro-2-(6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole- 4-methyl formate:
Figure PCTCN2016087368-appb-000073
Figure PCTCN2016087368-appb-000073
按照实施例2中步骤1的操作方法，以6-氟-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(100mg，0.30mmol)为原料，经甲基化得浅黄色固体77mg，产率为74％(二氯甲烷：甲醇＝30：1)。According to the procedure of Step 1 in Example 2, 6-fluoro-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[ d] Methyl imidazole-4-carboxylate (100 mg, 0.30 mmol) was obtained as a material.
1H NMR(300MHz,CD3OD)δ7.68(s,1H),7.54(dd,J＝9.9,2.5Hz,1H),7.48(dd,J＝8.7,2.5Hz,1H),4.01(s,3H),3.76(s,2H),2.86(brs,4H),2.54(s,3H)；MS(ESI)346[M+H]+。 1 H NMR (300 MHz, CD 3 OD) δ 7.68 (s, 1H), 7.54 (dd, J = 9.9, 2.5 Hz, 1H), 7.48 (dd, J = 8.7, 2.5 Hz, 1H), 4.01 (s) , 3H), 3.76 (s, 2H), 2.86 (brs, 4H), 2.54 (s, 3H); MS (ESI) 346 [M+H] + .
步骤2. 6-氟-2-(6-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-16)：Step 2. 6-Fluoro-2-(6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole- 4-formamide (I-16):
Figure PCTCN2016087368-appb-000074
Figure PCTCN2016087368-appb-000074
按照制备化合物I-1的操作方法，以6-氟-2-(6-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(77mg，0.22mmol)为原料氨解得到灰白色粉末36mg，产率为55％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 6-fluoro-2-(6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H Methyl benzo[d]imidazole-4-carboxylate (77 mg, 0.22 mmol) was obtained as a crude material (yield: 36 mg, yield: 55% (dichloromethane:methanol = 10:1).
1H NMR(500MHz,DMSO-d6)δ9.08(brs,1H),7.94(s,1H),7.68(brs,1H),7.59–7.52(m,2H),3.60(s,2H),2.73(t,J＝5.3Hz,2H),2.67(t,J＝5.6Hz,2H),2.39(s,3H)；13C NMR(125MHz,DMSO-d6)δ165.49,158.72(d,J＝235.6Hz,C-F),149.43,137.74,136.05,135.16,129.23,128.59,122.98,110.32(d,J＝26.5Hz,CH-CF),101.83,53.72,51.99,45.40,25.65；HRMS(ESI):m/z Calcd.For C16H16FN4OS[M+H]+:331.1029；Found:331.1020。 1 H NMR (500MHz, DMSO- d 6) δ9.08 (brs, 1H), 7.94 (s, 1H), 7.68 (brs, 1H), 7.59-7.52 (m, 2H), 3.60 (s, 2H), 2.73 (t, J = 5.3 Hz, 2H), 2.67 (t, J = 5.6 Hz, 2H), 2.39 (s, 3H); 13 C NMR (125 MHz, DMSO-d 6 ) δ 165.49, 158.72 (d, J = 235.6 Hz, CF), 149.43, 137.74, 136.05, 135.16, 129.23, 128.59, 122.98, 110.32 (d, J = 26.5 Hz, CH-CF), 101.83, 53.72, 51.99, 45.40, 25.65; HRMS (ESI): m /z Calcd.For C 16 H 16 FN 4 OS [M+H] + : 331.1029; Found: 331.1020.
实施例17. 6-氟-2-(7-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-17的制备：Example 17. 6-Fluoro-2-(7-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole Preparation of 4-carboxamide, I-17:
步骤1. N-(2-(噻吩-3-基)乙基)乙酰胺： Step 1. N-(2-(Thien-3-yl)ethyl)acetamide:
Figure PCTCN2016087368-appb-000075
Figure PCTCN2016087368-appb-000075
0℃下，向溶有2-(噻吩-3-基)乙基胺盐酸盐(492mg，3.0mmol)与三乙胺(9.0mmol)的二氯甲烷溶液中缓慢滴加乙酰氯(283mg，3.6mmol)，室温搅拌30分钟。反应液用二氯甲烷萃取(50mL×3)，有机层依次用水洗，饱和食盐水洗，保留有机层，用无水硫酸钠干燥，过滤减压浓缩得浅黄色油状物497mg，产率为98％，直接进入下一步反应。To a solution of 2-(thien-3-yl)ethylamine hydrochloride (492 mg, 3.0 mmol) and triethylamine (9.0 mmol) in dichloromethane, acetyl chloride (283 mg, 3.6 mmol), stirred at room temperature for 30 minutes. The reaction mixture was extracted with methylene chloride (50 mL×3). EtOAcjjjjjjjjj Go directly to the next step.
1H NMR(300MHz,Chloroform-d)δ7.32–7.32(m,1H),7.01(s,1H),6.95(d,J＝4.6Hz,1H),5.02(brs,1H),3.55–3.46(m,2H),2.85(t,J＝6.9Hz,2H),1.95(s,3H)；MS(ESI)170[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 7.32 - 7.32 (m, 1H), 7.01 (s, 1H), 6.95 (d, J = 4.6 Hz, 1H), 5.02 (brs, 1H), 3.55 - 3.46 (m, 2H), 2.85 (t, J = 6.9 Hz, 2H), 1.95 (s, 3H); MS (ESI) 170 [M+H] + .
步骤2. 7-甲基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 2. 7-Methyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000076
Figure PCTCN2016087368-appb-000076
将N-(2-(噻吩-3-基)乙基)乙酰胺(497mg，2.9mmol)溶于3mL三氯氧磷中，加热至60℃，反应两小时，冷却，将反应液倒入至冰水中，用饱和Na2CO3溶液中和至中性，用乙酸乙酯萃取(50mL×3)，有机层依次用水洗，饱和食盐水洗，保留有机层，用无水硫酸钠干燥，过滤减压浓缩得到黄色油状物，将该油状物溶于5mL无水甲醇中，0℃下，加入NaBH4(220mg，5.8mmol)，室温搅拌1小时，反应液用乙酸乙酯萃取(50mL×3)，有机层依次用水洗，饱和食盐水洗，保留有机层，用无水硫酸钠干燥，过滤减压浓缩得到黄色油状物，将该油状物溶于5mL二氯甲烷中，0℃下，加入二碳酸二叔丁酯(632mg，2.9mmol)，室温搅拌30分钟，减压浓缩得粗产品，经柱层析(石油醚：乙酸乙酯＝30：1)得到无色油状物506mg，三步总产率为69％。N-(2-(thien-3-yl)ethyl)acetamide (497 mg, 2.9 mmol) was dissolved in 3 mL of phosphorus oxychloride, heated to 60 ° C, reacted for two hours, cooled, and the reaction solution was poured into The mixture was neutralized with a saturated Na 2 CO 3 solution to dryness, and extracted with ethyl acetate (50 mL×3). The organic layer was washed successively with water and brine, and the organic layer was dried. concentrated under pressure to give a yellow oil. the oil was dissolved in 5mL of anhydrous methanol at 0 deg.] C, was added NaBH 4 (220mg, 5.8mmol), stirred for 1 hour at room temperature, the reaction solution was extracted with ethyl acetate (50mL × 3) The organic layer was washed successively with water and brine, and brine, evaporated, evaporated, evaporated, evaporated Di-tert-butyl ester (632 mg, 2.9 mmol), stirred at room temperature for 30 min, EtOAc (EtOAc) The rate is 69%.
1H NMR(300MHz,Chloroform-d)δ7.13(d,J＝5.0Hz,1H),6.75(d,J＝5.1Hz,1H),5.28(s,1H),4.30(s,1H),3.03(t,J＝12.6Hz,1H),2.77–2.54(m,2H),1.48(s,9H),1.46(d,J＝6.7Hz,3H)；MS(ESI)276[M+Na]+。 1 H NMR (300MHz, Chloroform- d) δ7.13 (d, J = 5.0Hz, 1H), 6.75 (d, J = 5.1Hz, 1H), 5.28 (s, 1H), 4.30 (s, 1H), 3.03 (t, J = 12.6 Hz, 1H), 2.77 - 2.54 (m, 2H), 1.48 (s, 9H), 1.46 (d, J = 6.7 Hz, 3H); MS (ESI) 276 [M+Na] + .
步骤3. 2-溴-7-甲基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 3. 2-Bromo-7-methyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000077
Figure PCTCN2016087368-appb-000077
按照实施例1中步骤2的操作方式，以7-甲基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(506mg，2.0mmol)为原料经溴代制得465mg油状物，产率为70％(石油醚：乙酸乙酯＝30：1)。Following the procedure of Step 2 in Example 1, 7-methyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (506 mg, 2.0 mmol) 465 mg of an oil was obtained by bromination of the starting material in a yield of 70% ( petroleum ether: ethyl acetate = 30:1).
1H NMR(300MHz,Chloroform-d)δ6.71(s,1H),5.16(brs,1H),4.25(brs,1H), 3.00(t,J＝11.0Hz,1H),2.73–2.43(m,2H),1.48(s,9H),1.41(d,J＝6.7Hz,3H)；MS(ESI)354[M+Na]+。 1 H NMR (300 MHz, Chloroform-d) δ 6.71 (s, 1H), 5.16 (brs, 1H), 4.25 (brs, 1H), 3.00 (t, J = 11.0 Hz, 1H), 2.73 - 2.43 (m) , 2H), 1.48 (s, 9H), 1.41 (d, J = 6.7 Hz, 3H); MS (ESI) 354 [M+Na] + .
步骤4. 2-甲酰基-7-甲基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 4. 2-Formyl-7-methyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000078
Figure PCTCN2016087368-appb-000078
按照实施例1中步骤3的操作方法，以2-溴-7-甲基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(12-3)(465mg，1.4mmol)为原料，用叔丁基锂正己烷溶液(1.6M)代替正丁基锂四氢呋喃溶液，制备得到淡黄色油状物295mg，产率为75％(石油醚：乙酸乙酯＝8：1)。Following the procedure of Step 3 in Example 1, 2-bromo-7-methyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (12 -3) (465 mg, 1.4 mmol) was used as a starting material, and a solution of n-butyllithium tetrahydrofuran was used to replace the n-butyllithium tetrahydrofuran solution to obtain 295 mg of a pale yellow oil (yield: 75%). Ethyl ester = 8:1).
1H NMR(300MHz,Chloroform-d)δ9.83(s,1H),7.44(s,1H),5.33(brs,1H),4.33(brs,1H),3.02(t,J＝12.8Hz,1H),2.70(m,2H),1.49(d,J＝6.7Hz,3H),1.49(s,9H)；MS(ESI)304[M+Na]+。 1 H NMR (300 MHz, Chloroform-d) δ 9.83 (s, 1H), 7.44 (s, 1H), 5.33 (brs, 1H), 4.33 (brs, 1H), 3.02 (t, J = 12.8 Hz, 1H) ), 2.70 (m, 2H), 1.49 (d, J = 6.7 Hz, 3H), 1.49 (s, 9H); MS (ESI) 304 [M+Na] + .
步骤5. 2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-7-甲基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 5. 2-(6-Fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-7-methyl-4,7-dihydrothieno[2,3 -c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000079
Figure PCTCN2016087368-appb-000079
按照实施例1中步骤4的操作方法，以2-甲酰基-7-甲基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(2-3)(295mg，1.05mmol)和2,3-二氨基-5-氟苯甲酸甲酯(193mg，1.05mmol)为原料制备得到淡黄色粉末262mg，产率为59％(石油醚：乙酸乙酯＝3：1)。According to the procedure of Step 4 in Example 1, 2-formyl-7-methyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester ( 2-3) (295 mg, 1.05 mmol) and 2,3-diamino-5-fluorobenzoic acid methyl ester (193 mg, 1.05 mmol) were used as starting materials to obtain 262 mg of pale yellow powder in a yield of 59% ( petroleum ether: acetic acid Ethyl ester = 3:1).
1H NMR(300MHz,Chloroform-d)δ7.63(dd,J＝8.9,2.5Hz,1H),7.59(dd,J＝9.5,2.5Hz,1H),7.35(s,1H),5.34(brs,1H),4.34(brs,1H),4.02(s,3H),3.06(t,J＝11.2Hz,1H),2.86–2.54(m,1H),1.51(d,J＝6.8Hz,3H),1.51(s,9H)；MS(ESI)446[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 7.63 (dd, J = 8.9, 2.5 Hz, 1H), 7.59 (dd, J = 9.5, 2.5 Hz, 1H), 7.35 (s, 1H), 5.34 (brs) , 1H), 4.34 (brs, 1H), 4.02 (s, 3H), 3.06 (t, J = 11.2 Hz, 1H), 2.86 - 2.54 (m, 1H), 1.51 (d, J = 6.8 Hz, 3H) , 1.51 (s, 9H); MS (ESI) 446 [M+H] + .
步骤6. 6-氟-2-(7-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 6. 6-Fluoro-2-(7-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole- 4-methyl formate:
Figure PCTCN2016087368-appb-000080
Figure PCTCN2016087368-appb-000080
按照实施例1中步骤5的操作方法，以2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2- 基)-7-甲基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(3-7)(262mg，0.59mmol)为原料去保护基得到白色粉末123mg，产率为60％(二氯甲烷：甲醇＝50：1)。Following the procedure of Step 5 in Example 1, 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazole-2- Base - 7-methyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (3-7) (262 mg, 0.59 mmol) as a starting material for protection The base obtained 123 mg of a white powder, yield 60% (dichloromethane:methanol = 50:1).
1H NMR(300MHz,Methanol-d4)δ7.64(s,1H),7.53(dd,J＝9.9,2.4Hz,1H),7.47(dd,J＝8.8,2.5Hz,1H),4.14(q,J＝6.9Hz,1H),4.00(s,3H),3.32–3.28(m,1H),2.97(ddd,J＝12.8,9.8,5.1Hz,1H),2.86–2.60(m,2H),1.49(d,J＝6.7Hz,3H)；MS(ESI)346[M+H]+。 1 H NMR (300MHz, Methanol- d 4) δ7.64 (s, 1H), 7.53 (dd, J = 9.9,2.4Hz, 1H), 7.47 (dd, J = 8.8,2.5Hz, 1H), 4.14 ( q, J = 6.9 Hz, 1H), 4.00 (s, 3H), 3.32 - 3.28 (m, 1H), 2.97 (ddd, J = 12.8, 9.8, 5.1 Hz, 1H), 2.86 - 2.60 (m, 2H) , 1.49 (d, J = 6.7 Hz, 3H); MS (ESI) 346 [M+H] + .
步骤7. 6-氟-2-(7-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-17)：Step 7. 6-Fluoro-2-(7-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole- 4-carboxamide (I-17):
Figure PCTCN2016087368-appb-000081
Figure PCTCN2016087368-appb-000081
按照制备化合物I-1的操作方法，以6-氟-2-(7-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(4-7)(123mg，0.35mmol)为原料氨解得到淡黄色粉末102mg，产率为87％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 6-fluoro-2-(7-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H - Benzo[d]imidazole-4-carboxylic acid methyl ester (4-7) (123 mg, 0.35 mmol) was obtained as a crude material to give a pale yellow powder (yield: 87%, yield: 87% (dichloromethane:methanol = 10:1) .
1H NMR(300MHz,DMSO-d6)δ8.96(brs,1H),7.90(brs,1H),7.68(s,1H),7.54(dd,J＝6.3,3.0Hz,1H),7.51(dd,J＝4.4,3.0Hz,1H),4.17(q,J＝6.6Hz,2H),3.28-3.17(m,1H),2.99-2.86(m,1H),2.73-2.58(m,2H),1.41(d,J＝6.6Hz,3H)；13C NMR(125MHz,DMSO-d6)δ165.88,158.92(d,J＝237.4Hz,C-F),149.34,143.87,135.76,129.20,129.02,122.86,110.32(d,J＝26.9Hz,CH-CF),102.04(d,J＝25.6Hz,CH-CF),50.37,41.89,25.74,22.74；HRMS(ESI):m/z Calcd.For C16H16FN4OS[M+H]+:331.1029；Found:331.1023. 1 H NMR (300MHz, DMSO- d 6) δ8.96 (brs, 1H), 7.90 (brs, 1H), 7.68 (s, 1H), 7.54 (dd, J = 6.3,3.0Hz, 1H), 7.51 ( Dd, J=4.4, 3.0 Hz, 1H), 4.17 (q, J=6.6 Hz, 2H), 3.28-3.17 (m, 1H), 2.99-2.86 (m, 1H), 2.73-2.58 (m, 2H) , 1.41 (d, J = 6.6 Hz, 3H); 13 C NMR (125MHz, DMSO-d 6 ) δ 165.88, 158.92 (d, J = 237.4 Hz, CF), 149.34, 143.87, 135.76, 129.20, 129.02, 122.86, 110.32 (d, J = 26.9 Hz, CH-CF), 102.04 (d, J = 25.6 Hz, CH-CF), 50.37, 41.89, 25.74, 22.74; HRMS (ESI): m/z Calcd. For C 16 H 16 FN 4 OS[M+H] + : 331.1029; Found: 331.1023.
实施例18. 2-(7-乙基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺，I-18的制备：Example 18. 2-(7-Ethyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H-benzo[d]imidazole Preparation of 4-carboxamide, I-18:
步骤1. N-(2-(噻吩-3-基)乙基)丙酰胺：Step 1. N-(2-(Thien-3-yl)ethyl)propanamide:
Figure PCTCN2016087368-appb-000082
Figure PCTCN2016087368-appb-000082
按照制备16-1的操作方法，以2-(噻吩-3-基)乙基胺盐酸盐(492mg，3.0mmol)，丙酰氯(333mg，3.6mmol)为原料制得浅黄色油状物543mg，产率为99％，直接进入下一步反应。543 mg of pale yellow oil was obtained from 2-(thiophen-3-yl)ethylamine hydrochloride (492 mg, 3.0 mmol), propionyl chloride (333 mg, 3.6 mmol). The yield was 99% and proceeded directly to the next reaction.
1H NMR(300MHz,Chloroform-d)δ7.29(dd,J＝5.0,2.9Hz,1H),7.02–6.99(m,1H),6.95(dd,J＝4.9,1.3Hz,1H),5.46(brs,1H),3.52(dd,J＝12.8,6.8Hz,2H),2.85(t,J＝6.9Hz,2H),2.17(q,J＝7.6Hz,2H),1.13(t,J＝7.6Hz,3H)；MS(ESI)184[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 7.29 (dd, J = 5.0, 2.9 Hz, 1H), 7.02 - 6.99 (m, 1H), 6.95 (dd, J = 4.9, 1.3 Hz, 1H), 5.46 (brs, 1H), 3.52 (dd, J = 12.8, 6.8 Hz, 2H), 2.85 (t, J = 6.9 Hz, 2H), 2.17 (q, J = 7.6 Hz, 2H), 1.13 (t, J = 7.6 Hz, 3H); MS (ESI) 184 [M+H] + .
步骤2. 7-乙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 2. 7-Ethyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000083
Figure PCTCN2016087368-appb-000083
按照制备14-3的操作方法，以N-(2-(噻吩-3-基)乙基)丙酰胺(16-2)(543mg，3.0mmol)为原料制得无色油状物529mg，三步总产率为66％(石油醚：乙酸乙酯＝30：1)。According to the procedure of Preparation 14-3, N-(2-(thiophen-3-yl)ethyl)propanamide (16-2) (543 mg, 3.0 mmol) was used as the starting material to obtain 529 mg of colorless oil. The total yield was 66% (petroleum ether: ethyl acetate = 30:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ7.13(d,J＝5.0Hz,1H),6.76(d,J＝5.0Hz,1H),5.22(brs,1H),4.28–4.07(m,1H),3.15–2.89(m,2H),2.57–2.51(m,1H),1.90–1.74(m,2H),1.47(s,9H),1.03(t,J＝7.4Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ7.13(d,J＝5.0Hz,1H),6.76(d,J＝5.0Hz,1H),5.10(brs,1H),4.49–4.28(m,1H),2.86–2.64(m,2H),2.62–2.58(m,1H),1.90–1.74(m,2H),1.47(s,9H),1.03(t,J＝7.4Hz,3H)；MS(ESI)290[M+Na]+。Conformation isomer 1 : 1 H NMR (300 MHz, Chloroform-d) δ 7.13 (d, J = 5.0 Hz, 1H), 6.76 (d, J = 5.0 Hz, 1H), 5.22 (brs, 1H), 4.28 –4.07(m,1H), 3.15–2.89(m,2H), 2.57–2.51(m,1H), 1.90–1.74(m,2H), 1.47(s,9H),1.03(t,J=7.4Hz , 3H); conformational isomer 2: 1 H NMR (300 MHz, Chloroform-d) δ 7.13 (d, J = 5.0 Hz, 1H), 6.76 (d, J = 5.0 Hz, 1H), 5.10 (brs, 1H), 4.49–4.28 (m, 1H), 2.86–2.64 (m, 2H), 2.62–2.58 (m, 1H), 1.90–1.74 (m, 2H), 1.47 (s, 9H), 1.03 (t, J = 7.4 Hz, 3H); MS (ESI) 290 [M+Na] + .
步骤3. 2-溴-7-乙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 3. 2-Bromo-7-ethyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000084
Figure PCTCN2016087368-appb-000084
按照制备12-3的操作方式，以7-乙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(14-4)(529mg，2.0mmol)为原料经溴代制得408mg白色固体，产率为60％(石油醚：乙酸乙酯＝30：1)。According to the procedure of Preparation 12-3, 4-ethyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (14-4) (529 mg, 2.0 mmol) was obtained by bromination of 408 mg of a white solid (yield: 60% ( petroleum ether: ethyl acetate = 30:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ6.72(s,1H),5.06(brs,1H),4.23–4.00(m,1H),3.19–2.86(m,2H),2.45(d,J＝3.9Hz,1H),1.77(p,J＝7.3Hz,2H),1.47(s,9H),1.00(t,J＝7.4Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ6.72(s,1H),4.97(brs,1H),4.46–4.23(m,1H),4.23–4.00(m,1H),2.80–2.55(m,2H),2.51(d,J＝4.1Hz,1H),1.77(p,J＝7.3Hz,2H),1.47(s,9H),1.00(t,J＝7.4Hz,3H)；MS(ESI)368[M+Na]+。Conformation isomer 1 : 1 H NMR (300MHz, Chloroform-d) δ 6.72 (s, 1H), 5.06 (brs, 1H), 4.23 - 4.00 (m, 1H), 3.19 - 2.86 (m, 2H), 2.45 (d, J = 3.9 Hz, 1H), 1.77 (p, J = 7.3 Hz, 2H), 1.47 (s, 9H), 1.00 (t, J = 7.4 Hz, 3H); conformation 2: 1 H NMR (300MHz, Chloroform-d) δ 6.72 (s, 1H), 4.97 (brs, 1H), 4.46 - 4.23 (m, 1H), 4.23 - 4.00 (m, 1H), 2.80 - 2.55 (m, 2H) ), 2.51 (d, J = 4.1 Hz, 1H), 1.77 (p, J = 7.3 Hz, 2H), 1.47 (s, 9H), 1.00 (t, J = 7.4 Hz, 3H); MS (ESI) 368 [M+Na] + .
步骤4. 7-乙基-2-甲酰基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 4. 7-Ethyl-2-formyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000085
Figure PCTCN2016087368-appb-000085
按照制备化合物2-3的操作方法，以2-溴-7-乙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(12-4)(408mg，1.2mmol)为原料制备得到淡黄色油状物329mg，产率为93％(石油醚：乙酸乙酯＝8：1)。 According to the procedure for the preparation of compound 2-3, 2-bromo-7-ethyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (12- 4) (408 mg, 1.2 mmol) was obtained as a crude material (yield: 329 mg,yield: 93%) (yield: petroleum ether: ethyl acetate = 8:1).
1H NMR(300MHz,Chloroform-d)δ9.84(s,1H),7.45(s,1H),5.40–5.01(m,1H),4.60–4.10(m,1H),3.17–2.51(m,3H),1.96–1.75(m,2H),1.48(d,J＝1.6Hz,9H),1.04(t,J＝7.4Hz,3H)；MS(ESI)318[M+Na]+。 1 H NMR (300 MHz, Chloroform-d) δ 9.84 (s, 1H), 7.45 (s, 1H), 5.40 - 5.01 (m, 1H), 4.60 - 4.10 (m, 1H), 3.17 - 2.51 (m, 3H), 1.96 - 1.75 (m, 2H), 1.48 (d, J = 1.6 Hz, 9H), 1.04 (t, J = 7.4 Hz, 3H); MS (ESI) 318 [M+Na] + .
步骤5. 7-乙基-2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 5. 7-Ethyl-2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7-dihydrothieno[2,3 -c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000086
Figure PCTCN2016087368-appb-000086
按照制备化合物3-7的操作方法，以7-乙基-2-甲酰基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(2-4)(329mg，1.12mmol)和2,3-二氨基-5-氟苯甲酸甲酯(206mg，1.12mmol)为原料制备得到淡黄色粉末329mg，产率为64％(石油醚：乙酸乙酯＝3：1)。According to the procedure for the preparation of compound 3-7, 7-ethyl-2-formyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (2 -4) (329 mg, 1.12 mmol) and 2,3-diamino-5-fluorobenzoic acid methyl ester (206 mg, 1.12 mmol) were used as starting materials to give 339 mg of pale yellow powder, yield 64% ( petroleum ether: acetic acid Ester = 3:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ10.41(s,1H),7.63(dd,J＝8.9,2.5Hz,1H),7.59(dd,J＝9.7,2.4Hz,1H),7.36(s,1H),5.30(brs,1H),4.33–4.20(m,1H),4.01(s,3H),3.13–2.94(m,2H),2.64–2.55(m,1H),1.94–1.78(m,2H),1.74–1.56(m,2H),1.07(t,J＝7.4Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ10.41(s,1H),7.63(dd,J＝8.9,2.5Hz,1H),7.59(dd,J＝9.7,2.4Hz,1H),7.36(s,1H),5.16(brs,1H),4.52–4.36(m,1H),4.01(s,3H),2.86–2.71(m,2H),2.68–2.64(m,1H),1.94–1.78(m,2H),1.74–1.56(m,2H),1.07(t,J＝7.4Hz,3H)；MS(ESI)460[M+H]+。Conformation isomer 1 : 1 H NMR (300 MHz, Chloroform-d) δ 10.41 (s, 1H), 7.63 (dd, J = 8.9, 2.5 Hz, 1H), 7.59 (dd, J = 9.7, 2.4 Hz, 1H), 7.36 (s, 1H), 5.30 (brs, 1H), 4.33 - 4.20 (m, 1H), 4.01 (s, 3H), 3.13 - 2.94 (m, 2H), 2.64 - 2.55 (m, 1H) , 1.94 - 1.78 (m, 2H), 1.74 - 1.56 (m, 2H), 1.07 (t, J = 7.4 Hz, 3H); conformation 2: 1 H NMR (300 MHz, Chloroform-d) δ 10.41 (s, 1H), 7.63 (dd, J = 8.9, 2.5 Hz, 1H), 7.59 (dd, J = 9.7, 2.4 Hz, 1H), 7.36 (s, 1H), 5.16 (brs, 1H), 4.52– 4.36 (m, 1H), 4.01 (s, 3H), 2.86 - 2.71 (m, 2H), 2.68 - 2.64 (m, 1H), 1.94 - 1.78 (m, 2H), 1.74 - 1.56 (m, 2H), 1.07 (t, J = 7.4 Hz, 3H); MS (ESI) 460 [M+H] + .
步骤6. 2-(7-乙基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯：Step 6. 2-(7-Ethyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H-benzo[d]imidazole- 4-methyl formate:
Figure PCTCN2016087368-appb-000087
Figure PCTCN2016087368-appb-000087
按照制备化合物4-7的操作方法，以7-乙基-2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(3-8)(329mg，0.72mmol)为原料去保护基得到白色粉末144mg，产率为56％(二氯甲烷：甲醇＝50：1)。According to the procedure for the preparation of compound 4-7, 7-ethyl-2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7- Dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (3-8) (329 mg, 0.72 mmol) was used as a starting material to afford the title compound 144 mg (yield: 56%) Dichloromethane: methanol = 50:1).
1H NMR(300MHz,Chloroform-d)δ10.40(brs,1H),7.62(dd,J＝9.2,1.3Hz,1H),7.59(dd,J＝9.5,1.3Hz,1H),7.37(s,1H),4.01(s,3H),3.36(ddd,J＝12.7,5.3,3.3Hz,1H),3.09–2.93(m,1H),2.82–2.59(m,2H),2.07–1.85(m,2H),1.10(t,J＝7.3Hz,3H)；MS(ESI)360[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 10.40 (brs, 1H), 7.62 (dd, J = 9.2, 1.3 Hz, 1H), 7.59 (dd, J = 9.5, 1.3 Hz, 1H), 7.37 (s) , 1H), 4.01 (s, 3H), 3.36 (ddd, J = 12.7, 5.3, 3.3 Hz, 1H), 3.09 - 2.93 (m, 1H), 2.82 - 2.59 (m, 2H), 2.07 - 1.85 (m , 2H), 1.10 (t, J = 7.3 Hz, 3H); MS (ESI) 360 [M+H] + .
步骤7. 2-(7-乙基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲 酰胺(I-18)：Step 7. 2-(7-Ethyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H-benzo[d]imidazole- 4-A Amide (I-18):
Figure PCTCN2016087368-appb-000088
Figure PCTCN2016087368-appb-000088
按照制备化合物I-1的操作方法，以2-(7-乙基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯(4-8)(144mg，0.40mmol)为原料氨解得到淡黄色粉末119mg，产率为86％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 2-(7-ethyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H - Benzo[d]imidazole-4-carboxylic acid methyl ester (4-8) (144 mg, 0.40 mmol) was obtained as a crude material (yield: 119 mg,yield: 86% (methylene chloride:methanol = 10:1) .
1H NMR(500MHz,DMSO-d6)δ9.00(brs,1H),7.97(brs,1H),7.78(s,1H),7.56(dd,J＝10.7,2.7Hz,1H),7.53(dd,J＝8.3,2.7Hz,1H),4.23(t,J＝6.0Hz,1H),3.34(dt,J＝12.4,4.1Hz,1H),3.06(dt,J＝12.4,4.1Hz,1H),2.87–2.71(m,2H),1.99-1.90(m,1H),1.89–1.76(m,1H),1.08(t,J＝7.3Hz,3H)；13C NMR(125MHz,DMSO-d6)δ165.56,158.71(d,J＝238.1Hz,C-F),149.09,140.19,135.63,130.15,128.89,122.95,110.39(d,J＝26.3Hz,CH-CF),102.48(d,J＝27.1Hz,CH-CF),55.63,41.39,28.94,24.58,10.69；HRMS(ESI):m/z Calcd.For C17H18FN4OS[M]+:345.1185；Found:345.1178。 1 H NMR (500MHz, DMSO- d 6) δ9.00 (brs, 1H), 7.97 (brs, 1H), 7.78 (s, 1H), 7.56 (dd, J = 10.7,2.7Hz, 1H), 7.53 ( Dd, J = 8.3, 2.7 Hz, 1H), 4.23 (t, J = 6.0 Hz, 1H), 3.34 (dt, J = 12.4, 4.1 Hz, 1H), 3.06 (dt, J = 12.4, 4.1 Hz, 1H) ), 2.87 - 2.71 (m, 2H), 1.99-1.90 (m, 1H), 1.89 - 1.76 (m, 1H), 1.08 (t, J = 7.3 Hz, 3H); 13 C NMR (125 MHz, DMSO-d) 6 ) δ165.56,158.71 (d, J=238.1 Hz, CF), 149.09, 140.19, 135.63, 130.15, 128.89, 122.95, 110.39 (d, J=26.3 Hz, CH-CF), 102.48 (d, J=27.1 Hz) , CH-CF), 55.63, 41.39, 28.94, 24.58, 10.69; HRMS (ESI): m/z Calcd. For C 17 H 18 FN 4 OS [M] + : 345.1185; Found: 345.1178.
实施例19. 6-氟-2-(7-丙基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-19的制备：Example 19. 6-Fluoro-2-(7-propyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole Preparation of 4-carboxamide, I-19:
步骤1. N-(2-(噻吩-3-基)乙基)丁酰胺：Step 1. N-(2-(Thien-3-yl)ethyl)butanamide:
Figure PCTCN2016087368-appb-000089
Figure PCTCN2016087368-appb-000089
按照制备16-1的操作方法，以2-(噻吩-3-基)乙基胺盐酸盐(492mg，3.0mmol)，丁酰氯(383mg，3.6mmol)为原料制得浅黄色油状物585mg，产率为99％，直接进入下一步反应。585 mg of light yellow oil was obtained from 2-(thiophen-3-yl)ethylamine hydrochloride (492 mg, 3.0 mmol), butyryl chloride (383 mg, 3.6 mmol). The yield was 99% and proceeded directly to the next reaction.
1H NMR(300MHz,Chloroform-d)δ7.32–7.28(m,1H),7.02–6.98(m,1H),6.95(d,J＝4.8Hz,1H),5.43(brs,1H),3.57–3.48(m,2H),2.85(t,J＝6.8Hz,2H),2.11(dd,J＝8.3,6.7Hz,2H),1.70–1.60(m,2H),0.92(t,J＝7.4Hz,3H)；MS(ESI)198[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 7.32 - 7.28 (m, 1H), 7.02 - 6.98 (m, 1H), 6.95 (d, J = 4.8 Hz, 1H), 5.43 (brs, 1H), 3.57 –3.48(m,2H), 2.85 (t, J=6.8Hz, 2H), 2.11 (dd, J=8.3, 6.7Hz, 2H), 1.70–1.60 (m, 2H), 0.92 (t, J=7.4) Hz, 3H); MS (ESI) 198 [M+H] + .
步骤2. 7-丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 2. 7-Propyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000090
Figure PCTCN2016087368-appb-000090
按照制备14-3的操作方法，以N-(2-(噻吩-3-基)乙基)丁酰胺(16-3)(585mg，3.0 mmol)为原料制得无色油状物532mg，三步总产率为63％(石油醚：乙酸乙酯＝30：1)。According to the procedure of Preparation 14-3, N-(2-(thien-3-yl)ethyl)butanamide (16-3) (585 mg, 3.0) Methyl) 532 mg of a colorless oil was obtained from the crude material. The crude product was obtained in three steps ( petroleum ether: ethyl acetate = 30:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ7.13(d,J＝5.1Hz,1H),6.76(d,J＝5.1Hz,1H),5.29(brs,1H),4.25–4.09(m,1H),3.18–2.91(m,2H),2.54(d,J＝4.2Hz,1H),1.84–1.64(m,2H),1.55–1.42(m,2H),1.47(s,9H),0.96(t,J＝7.3Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ7.13(d,J＝5.1Hz,1H),6.76(d,J＝5.1Hz,1H),5.17(brs,1H),4.44–4.30(m,1H),2.86–2.65(m,2H),2.59(d,J＝3.9Hz,1H),1.84–1.64(m,2H),1.55–1.42(m,2H),1.47(s,9H),0.96(t,J＝7.3Hz,3H)；MS(ESI)304[M+Na]+。Conformation isomer 1 : 1 H NMR (300 MHz, Chloroform-d) δ 7.13 (d, J = 5.1 Hz, 1H), 6.76 (d, J = 5.1 Hz, 1H), 5.29 (brs, 1H), 4.25 –4.09 (m,1H), 3.18–2.91 (m, 2H), 2.54 (d, J=4.2 Hz, 1H), 1.84–1.64 (m, 2H), 1.55–1.42 (m, 2H), 1.47 (s) , 9H), 0.96 (t, J = 7.3 Hz, 3H); conformational isomer 2: 1 H NMR (300 MHz, Chloroform-d) δ 7.13 (d, J = 5.1 Hz, 1H), 6.76 (d, J=5.1 Hz, 1H), 5.17 (brs, 1H), 4.44–4.30 (m, 1H), 2.86–2.65 (m, 2H), 2.59 (d, J=3.9 Hz, 1H), 1.84–1.64 (m) , 2H), 1.55 - 1.42 (m, 2H), 1.47 (s, 9H), 0.96 (t, J = 7.3 Hz, 3H); MS (ESI) 304 [M+Na] + .
步骤3. 2-溴-7-丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 3. 2-Bromo-7-propyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000091
Figure PCTCN2016087368-appb-000091
按照制备12-3的操作方式，以7-丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(14-5)(532mg，1.9mmol)为原料经溴代制得512mg无色晶体，产率为76％(石油醚：乙酸乙酯＝30：1)。According to the procedure of Preparation 12-3, 4-propyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (14-5) (532 mg, 1.9 mmol) was obtained by bromination of 512 mg of colorless crystals, yield 76% ( petroleum ether: ethyl acetate = 30:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ6.71(s,1H),5.24–5.10(m,1H),4.21–4.03(m,1H),3.18–2.84(m,2H),2.49–2.37(m,1H),1.85–1.61(m,2H),1.57–1.39(m,2H),1.47(s,9H),0.95(t,J＝7.3Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ6.71(s,1H),5.11–4.95(m,1H),4.42–4.26(m,1H),2.84–2.56(m,2H),2.54–2.47(m,1H),1.85–1.61(m,2H),1.57–1.39(m,2H),1.47(s,9H),0.95(t,J＝7.3Hz,3H)；MS(ESI)382[M+Na]+。Conformation isomer 1 : 1 H NMR (300 MHz, Chloroform-d) δ 6.71 (s, 1H), 5.24 - 5.10 (m, 1H), 4.21 - 4.03 (m, 1H), 3.18 - 2.84 (m, 2H) ), 2.49–2.37 (m, 1H), 1.85–1.61 (m, 2H), 1.57–1.39 (m, 2H), 1.47 (s, 9H), 0.95 (t, J = 7.3 Hz, 3H); Construct 2: 1 H NMR (300 MHz, Chloroform-d) δ 6.71 (s, 1H), 5.11 - 4.95 (m, 1H), 4.42 - 4.26 (m, 1H), 2.84 - 2.56 (m, 2H), 2.54–2.47 (m, 1H), 1.85–1.61 (m, 2H), 1.57–1.39 (m, 2H), 1.47 (s, 9H), 0.95 (t, J = 7.3 Hz, 3H); MS (ESI) 382 [M+Na] + .
步骤4. 2-甲酰基-7-丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 4. 2-Formyl-7-propyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000092
Figure PCTCN2016087368-appb-000092
按照制备化合物2-3的操作方法，以2-溴-7-丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(12-5)(512mg，1.4mmol)为原料制备得到淡黄色油状物426mg，产率为96％(石油醚：乙酸乙酯＝8：1)。According to the procedure for the preparation of compound 2-3, 2-bromo-7-propyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (12- 5) (512 mg, 1.4 mmol) was obtained as a crude material (yield: 426 mg,yield: 96%) (yield: petroleum ether: ethyl acetate = 8:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ9.83(s,1H),7.44(s,1H),5.49–5.29(m,1H),4.30–4.16(m,1H),3.18–2.90(m,2H),2.63–2.52(m,1H),1.91–1.64(m,2H),1.48(s,9H),0.97(t,J＝7.3Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ9.83(s,1H),7.44(s,1H),5.29–5.10(m,1H),4.50–4.34(m,1H), 2.87–2.69(m,2H),2.68–2.61(m,1H),1.91–1.64(m,2H),1.48(s,9H),0.97(t,J＝7.3Hz,3H)；MS(ESI)332[M+Na]+。Conformation isomer 1 : 1 H NMR (300 MHz, Chloroform-d) δ 9.83 (s, 1H), 7.44 (s, 1H), 5.49 - 5.29 (m, 1H), 4.30 - 4.16 (m, 1H), 3.18–2.90 (m, 2H), 2.63–2.52 (m, 1H), 1.91–1.64 (m, 2H), 1.48 (s, 9H), 0.97 (t, J = 7.3 Hz, 3H); conformational isomer 2: 1 H NMR (300 MHz, Chloroform-d) δ 9.83 (s, 1H), 7.44 (s, 1H), 5.29 - 5.10 (m, 1H), 4.50 - 4.34 (m, 1H), 2.87 - 2.69 ( m, 2H), 2.68 - 2.61 (m, 1H), 1.91 - 1.64 (m, 2H), 1.48 (s, 9H), 0.97 (t, J = 7.3 Hz, 3H); MS (ESI) 332 [M+ Na] + .
步骤5. 2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-7-丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 5. 2-(6-Fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-7-propyl-4,7-dihydrothieno[2,3 -c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000093
Figure PCTCN2016087368-appb-000093
按照制备化合物3-7的操作方法，以2-甲酰基-7-丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(2-5)(426mg，1.38mmol)和2,3-二氨基-5-氟苯甲酸甲酯(254mg，1.38mmol)为原料制备得到淡黄色粉末562mg，产率为86％(石油醚：乙酸乙酯＝3：1)。According to the procedure for the preparation of compound 3-7, 2-formyl-7-propyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (2 -5) (426 mg, 1.38 mmol) and 2,3-diamino-5-fluorobenzoic acid methyl ester (254 mg, 1.38 mmol) were used as starting materials to obtain 562 mg of pale yellow powder, yield 86% (petrole ether: acetic acid Ester = 3:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ10.42(s,1H),7.63(dd,J＝9.3,2.4Hz,1H),7.58(dd,J＝9.4,2.4Hz,1H),7.35(s,1H),5.36(brs,1H),4.31–4.16(m,1H),4.01(s,3H),2.88–2.68(m,2H),2.64–2.55(m,1H),1.90–1.72(m,4H),1.64–1.39(m,2H),1.49(s,9H),0.98(t,J＝7.3Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ10.42(s,1H),7.63(dd,J＝9.3,2.4Hz,1H),7.58(dd,J＝9.4,2.4Hz,1H),7.35(s,1H),5.21(brs,1H),4.50–4.34(m,1H),3.21–2.93(m,2H),2.69–2.61(m,1H),1.90–1.72(m,4H),1.64–1.39(m,2H),1.49(s,9H),0.98(t,J＝7.3Hz,3H)；MS(ESI)474[M+H]+。Conformational isomer 1: 1 H NMR (300 MHz, Chloroform-d) δ 10.42 (s, 1H), 7.63 (dd, J = 9.3, 2.4 Hz, 1H), 7.58 (dd, J = 9.4, 2.4 Hz, 1H), 7.35 (s, 1H), 5.36 (brs, 1H), 4.31 - 4.16 (m, 1H), 4.01 (s, 3H), 2.88 - 2.68 (m, 2H), 2.64 - 2.55 (m, 1H) , 1.90 - 1.72 (m, 4H), 1.64 - 1.39 (m, 2H), 1.49 (s, 9H), 0.98 (t, J = 7.3 Hz, 3H); conformation 2: 1 H NMR (300 MHz, Chloroform-d) δ 10.42 (s, 1H), 7.63 (dd, J = 9.3, 2.4 Hz, 1H), 7.58 (dd, J = 9.4, 2.4 Hz, 1H), 7.35 (s, 1H), 5.21. Brs, 1H), 4.50–4.34 (m, 1H), 3.21–2.93 (m, 2H), 2.69–2.61 (m, 1H), 1.90–1.72 (m, 4H), 1.64–1.39 (m, 2H), 1.49 (s, 9H), 0.98 (t, J = 7.3 Hz, 3H); MS (ESI) 474 [M+H] + .
步骤6. 6-氟-2-(7-丙基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 6. 6-Fluoro-2-(7-propyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole- 4-methyl formate:
Figure PCTCN2016087368-appb-000094
Figure PCTCN2016087368-appb-000094
按照制备化合物4-7的操作方法，以2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-7-丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(3-9)(329mg，0.72mmol)为原料去保护基得到淡黄色固体237mg，产率为88％(二氯甲烷：甲醇＝50：1)。According to the procedure for the preparation of compound 4-7, 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-7-propyl-4,7- Dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (3-9) (329 mg, 0.72 mmol) was used as the starting material to afford the residue to afford 237mg as pale yellow solid. (Dichloromethane: methanol = 50:1).
1H NMR(300MHz,Chloroform-d)δ10.39(brs,1H),7.62(dd,J＝8.8,2.4Hz,1H),7.58(dd,J＝9.7,2.4Hz,1H),7.38(s,1H),4.12–4.03(m,1H),4.02(s,3H),3.40–3.30(m,1H),3.09–2.93(m,1H),2.79–2.60(m,2H),1.95–1.69(m,2H),1.63–1.47(m,2H),1.00(t,J＝7.3Hz,3H)；MS(ESI)374[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ10.39 (brs, 1H), 7.62 (dd, J = 8.8,2.4Hz, 1H), 7.58 (dd, J = 9.7,2.4Hz, 1H), 7.38 (s , 1H), 4.12–4.03 (m, 1H), 4.02 (s, 3H), 3.40–3.30 (m, 1H), 3.09–2.93 (m, 1H), 2.79–2.60 (m, 2H), 1.95–1.69 (m, 2H), 1.63 - 1.47 (m, 2H), 1.00 (t, J = 7.3 Hz, 3H); MS (ESI) 374 [M+H] + .
步骤7. 6-氟-2-(7-丙基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲 酰胺(I-19)：Step 7. 6-Fluoro-2-(7-propyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole- 4-A Amide (I-19):
Figure PCTCN2016087368-appb-000095
Figure PCTCN2016087368-appb-000095
按照制备化合物I-1的操作方法，以6-氟-2-(7-丙基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(4-9)(237mg，0.63mmol)为原料氨解得到白色粉末137mg，产率为61％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 6-fluoro-2-(7-propyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H - Benzo[d]imidazole-4-carboxylic acid methyl ester (4-9) (237 mg, 0.63 mmol) was obtained as a crude material (yield: 137 mg,yield: 61% (dichloromethane:methanol = 10:1).
1H NMR(400MHz,DMSO-d6)δ8.99(brs,1H),7.93(brs,1H),7.69(s,1H),7.54(dd,J＝10.9,2.5Hz,1H),7.51(dd,J＝8.6,2.6Hz,1H),4.12(dd,J＝8.1,4.6Hz,1H),3.23(dt,J＝12.6,4.8Hz,1H),2.94(ddd,J＝13.2,8.9,5.4Hz,1H),2.78–2.57(m,2H),1.84-1.62(m,2H),1.59–1.41(m,2H),0.93(t,J＝7.3Hz,3H)；13C NMR(100MHz,DMSO-d6)δ165.47,158.64(d,J＝236.1Hz,C-F),149.21,142.22,135.84,129.42,128.92,122.91,110.29(d,J＝25.9Hz,CH-CF),102.02(d,J＝26.3Hz,CH-CF),54.22,41.62,38.79,25.44,19.09,14.31；HRMS(ESI):m/z Calcd.For C18H20FN4OS[M+H]+:359.1342；Found:359.1334。 1 H NMR (400MHz, DMSO- d 6) δ8.99 (brs, 1H), 7.93 (brs, 1H), 7.69 (s, 1H), 7.54 (dd, J = 10.9,2.5Hz, 1H), 7.51 ( Dd, J = 8.6, 2.6 Hz, 1H), 4.12 (dd, J = 8.1, 4.6 Hz, 1H), 3.23 (dt, J = 12.6, 4.8 Hz, 1H), 2.94 (ddd, J = 13.2, 8.9, 5.4 Hz, 1H), 2.78–2.57 (m, 2H), 1.84-1.62 (m, 2H), 1.59–1.41 (m, 2H), 0.93 (t, J = 7.3 Hz, 3H); 13 C NMR (100 MHz) , DMSO-d 6 ) δ 165.47, 158.64 (d, J = 236.1 Hz, CF), 149.21, 142.22, 135.84, 129.42, 128.92, 122.91, 110.29 (d, J = 25.9 Hz, CH-CF), 102.02 (d, J=26.3 Hz, CH-CF), 54.22, 41.62, 38.79, 25.44, 19.09, 14.31; HRMS (ESI): m/z Calcd. For C 18 H 20 FN 4 OS [M+H] + :359.1342;Found :359.1334.
实施例20. 2-(7-丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺，I-20的制备：Example 20. 2-(7-Butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H-benzo[d]imidazole Preparation of 4-carboxamide, I-20:
步骤1. N-(2-(噻吩-3-基)乙基)戊酰胺：Step 1. N-(2-(Thien-3-yl)ethyl)pentanamide:
Figure PCTCN2016087368-appb-000096
Figure PCTCN2016087368-appb-000096
按照制备16-1的操作方法，以2-(噻吩-3-基)乙基胺盐酸盐(492mg，3.0mmol)，戊酰氯(434mg，3.6mmol)为原料制得浅黄色油状物627mg，产率为99％，直接进入下一步反应。A pale yellow oil 627 mg was obtained from 2-(thiophen-3-yl)ethylamine hydrochloride (492 mg, 3.0 mmol), pentanoyl chloride (434 mg, 3.6 mmol). The yield was 99% and proceeded directly to the next reaction.
1H NMR(300MHz,Chloroform-d)δ7.32–7.27(m,1H),7.02–6.99(m,1H),6.98–6.94(m,1H),5.46(brs,1H),3.56–3.48(m,1H),3.03–2.93(m,1H),2.88–2.76(m,1H),2.16–2.09(m,1H),1.59–1.52(m,2H),1.37–1.28(m,2H),0.90(t,J＝7.3Hz,3H)；MS(ESI)212[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 7.32 - 7.27 (m, 1H), 7.02 - 6.99 (m, 1H), 6.98 - 6.94 (m, 1H), 5.46 (brs, 1H), 3.56 - 3.48 ( m, 1H), 3.03–2.93 (m, 1H), 2.88–2.76 (m, 1H), 2.16–2.09 (m, 1H), 1.59–1.52 (m, 2H), 1.37–1.28 (m, 2H), 0.90 (t, J = 7.3 Hz, 3H); MS (ESI) 212 [M+H] + .
步骤2. 7-丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯： Step 2. 7-Butyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000097
Figure PCTCN2016087368-appb-000097
按照制备14-3的操作方法，以N-(2-(噻吩-3-基)乙基)戊酰胺(16-4)(627mg，3.0mmol)为原料制得淡黄色油状物354mg，三步总产率为40％(石油醚：乙酸乙酯＝30：1)。According to the procedure of Preparation 14-3, N-(2-(thiophen-3-yl)ethyl)pentanamide (16-4) (627 mg, 3.0 mmol) The total yield was 40% (petroleum ether: ethyl acetate = 30:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ7.13(d,J＝5.0Hz,1H),6.76(d,J＝5.0Hz,1H),5.29(brs,1H),4.28–4.00(m,1H),3.16–2.87(m,2H),2.57–2.45(m,1H),1.86–1.67(m,2H),1.47(s,9H),1.45–1.21(m,4H),0.91(t,J＝6.8Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ7.13(d,J＝5.0Hz,1H),6.76(d,J＝5.0Hz,1H),5.16(brs,1H),4.48–4.28(m,1H),2.85–2.62(m,2H),2.62–2.57(m,1H),1.86–1.67(m,2H),1.47(s,9H),1.45–1.21(m,4H),0.91(t,J＝6.8Hz,3H)；MS(ESI)318[M+H]+。Conformation isomer 1 : 1 H NMR (300 MHz, Chloroform-d) δ 7.13 (d, J = 5.0 Hz, 1H), 6.76 (d, J = 5.0 Hz, 1H), 5.29 (brs, 1H), 4.28 – 4.00 (m, 1H), 3.16–2.87 (m, 2H), 2.57–2.45 (m, 1H), 1.86–1.67 (m, 2H), 1.47 (s, 9H), 1.45–1.21 (m, 4H) , 0.91 (t, J = 6.8 Hz, 3H); conformational isomer 2: 1 H NMR (300 MHz, Chloroform-d) δ 7.13 (d, J = 5.0 Hz, 1H), 6.76 (d, J = 5.0) Hz, 1H), 5.16 (brs, 1H), 4.48–4.28 (m, 1H), 2.85–2.62 (m, 2H), 2.62–2.57 (m, 1H), 1.86–1.67 (m, 2H), 1.47 ( s, 9H), 1.45 - 1.21 (m, 4H), 0.91 (t, J = 6.8 Hz, 3H); MS (ESI) 318 [M+H] + .
步骤3. 2-溴-7-丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 3. 2-Bromo-7-butyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000098
Figure PCTCN2016087368-appb-000098
按照制备12-3的操作方式，以7-丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(14-6)(354mg，1.2mmol)为原料经溴代制得347mg无色油状物，产率为77％(石油醚：乙酸乙酯＝30：1)。According to the procedure of Preparation 12-3, 4-butyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (14-6) (354 mg, 1.2 mmol) was obtained by bromination of 347 mg of a colorless oil, yield 77% ( petroleum ether: ethyl acetate = 30:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ6.71(s,1H),5.22–5.12(m,1H),4.21–4.04(m,1H),3.10–2.89(m,2H),2.45(d,J＝4.0Hz,1H),1.84–1.63(m,2H),1.47(s,9H),1.43–1.29(m,4H),0.91(t,J＝6.9Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ6.71(s,1H),5.09–4.94(m,1H),4.40–4.23(m,1H),2.75–2.56(m,2H),2.50(d,J＝3.3Hz,1H),1.84–1.63(m,2H),1.47(s,9H),1.43–1.29(m,4H),0.91(t,J＝6.9Hz,3H)；MS(ESI)396[M+Na]+。Conformation isomer 1 : 1 H NMR (300 MHz, Chloroform-d) δ 6.71 (s, 1H), 5.22 - 5.12 (m, 1H), 4.21 - 4.04 (m, 1H), 3.10 - 2.89 (m, 2H) ), 2.45 (d, J = 4.0 Hz, 1H), 1.84 - 1.63 (m, 2H), 1.47 (s, 9H), 1.43 - 1.29 (m, 4H), 0.91 (t, J = 6.9 Hz, 3H) Conformational isomer 2: 1 H NMR (300 MHz, Chloroform-d) δ 6.71 (s, 1H), 5.09 - 4.94 (m, 1H), 4.40 - 4.23 (m, 1H), 2.75 - 2.56 (m, 2H), 2.50 (d, J = 3.3 Hz, 1H), 1.84 - 1.63 (m, 2H), 1.47 (s, 9H), 1.43 - 1.29 (m, 4H), 0.91 (t, J = 6.9 Hz, 3H) MS (ESI) 396 [M+Na] + .
步骤4. 7-丁基-2-甲酰基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 4. 7-Butyl-2-formyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000099
Figure PCTCN2016087368-appb-000099
按照制备化合物2-3的操作方法，以2-溴-7-丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)- 羧酸叔丁酯(12-6)(347mg，0.9mmol)为原料制备得到无色油状物240mg，产率为81％(石油醚：乙酸乙酯＝8：1)。According to the procedure for the preparation of compound 2-3, 2-bromo-7-butyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)- Preparation of tert-butyl carboxylic acid tert-butyl ester (12-6) (347 mg, 0.9 mmol) afforded 240 mg (yield: 81%) (yield: petroleum ether: ethyl acetate = 8:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ9.83(s,1H),7.44(s,1H),5.42–5.28(m,1H),4.31–4.17(m,1H),3.15–2.90(m,2H),2.63–2.55(m,1H),1.79(dd,J＝6.5,5.1Hz,2H),1.48(s,9H),1.45–1.24(m,4H),0.92(t,J＝6.8Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ9.83(s,1H),7.44(s,1H),5.28–5.09(m,1H),4.50–4.33(m,1H),2.84–2.68(m,2H),2.69–2.61(m,1H),1.79(dd,J＝6.5,5.1Hz,2H),1.48(s,9H),1.45–1.24(m,4H),0.92(t,J＝6.8Hz,3H)；MS(ESI)346[M+Na]+。Conformation isomer 1 : 1 H NMR (300 MHz, Chloroform-d) δ 9.83 (s, 1H), 7.44 (s, 1H), 5.42 - 5.28 (m, 1H), 4.31 - 4.17 (m, 1H), 3.15–2.90 (m, 2H), 2.63–2.55 (m, 1H), 1.79 (dd, J=6.5, 5.1 Hz, 2H), 1.48 (s, 9H), 1.45–1.24 (m, 4H), 0.92 ( t, J = 6.8 Hz, 3H); conformational isomer 2: 1 H NMR (300 MHz, Chloroform-d) δ 9.83 (s, 1H), 7.44 (s, 1H), 5.28 - 5.09 (m, 1H) , 4.50–4.33 (m, 1H), 2.84–2.68 (m, 2H), 2.69–2.61 (m, 1H), 1.79 (dd, J=6.5, 5.1 Hz, 2H), 1.48 (s, 9H), 1.45 - 1.24 (m, 4H), 0.92 (t, J = 6.8 Hz, 3H); MS (ESI) 346 [M+Na] + .
步骤5. 7-丁基-2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 5. 7-Butyl-2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7-dihydrothieno[2,3 -c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000100
Figure PCTCN2016087368-appb-000100
按照制备化合物3-7的操作方法，以2-甲酰基-7-丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(2-6)(426mg，1.38mmol)和2,3-二氨基-5-氟苯甲酸甲酯(254mg，1.38mmol)为原料制备得到淡黄色粉末524mg，产率为78％(石油醚：乙酸乙酯＝3：1)。According to the procedure for the preparation of compound 3-7, 2-formyl-7-butyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (2 -6) (426 mg, 1.38 mmol) and 2,3-diamino-5-fluorobenzoic acid methyl ester (254 mg, 1.38 mmol) were used as starting materials to give 524 mg of pale yellow powder (yield: 78%). Ester = 3:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ10.42(brs,1H),7.64(dd,J＝8.9,2.5Hz,1H),7.61(dd,J＝9.8,2.5Hz,1H),7.35(s,1H),5.43–5.27(m,1H),4.32–4.17(m,1H),4.02(s,3H),3.21–2.92(m,2H),2.64–2.55(m,1H),1.83(q,J＝7.3Hz,2H),1.50(s,9H),1.47–1.20(m,4H),0.93(t,J＝7.0Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ10.42(brs,1H),7.64(dd,J＝8.9,2.5Hz,1H),7.61(dd,J＝9.8,2.5Hz,1H),7.35(s,1H),5.26–5.13(m,1H),4.50–4.33(m,1H),4.02(s,3H),2.87–2.70(m,2H),2.70–2.63(m,1H),1.83(q,J＝7.3Hz,2H),1.50(s,9H),1.47–1.20(m,4H),0.93(t,J＝7.0Hz,3H)；MS(ESI)488[M+H]+。Conformation isomer 1 : 1 H NMR (300 MHz, Chloroform-d) δ 10.42 (brs, 1H), 7.64 (dd, J = 8.9, 2.5 Hz, 1H), 7.61 (dd, J = 9.8, 2.5 Hz, 1H), 7.35 (s, 1H), 5.43 - 5.27 (m, 1H), 4.32 - 4.17 (m, 1H), 4.02 (s, 3H), 3.21 - 2.92 (m, 2H), 2.64 - 2.55 (m, 1H), 1.83 (q, J = 7.3 Hz, 2H), 1.50 (s, 9H), 1.47 - 1.20 (m, 4H), 0.93 (t, J = 7.0 Hz, 3H); conformation 2: 1 H NMR (300 MHz, Chloroform-d) δ 10.42 (brs, 1H), 7.64 (dd, J = 8.9, 2.5 Hz, 1H), 7.61 (dd, J = 9.8, 2.5 Hz, 1H), 7.35 (s, 1H), 5.26–5.13 (m, 1H), 4.50–4.33 (m, 1H), 4.02 (s, 3H), 2.87–2.70 (m, 2H), 2.70–2.63 (m, 1H), 1.83 (q, J = 7.3 Hz, 2H), 1.50 (s, 9H), 1.47 - 1.20 (m, 4H), 0.93 (t, J = 7.0 Hz, 3H); MS (ESI) 488 [M+H] + .
步骤6. 6-氟-2-(7-丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 6. 6-Fluoro-2-(7-butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole- 4-methyl formate:
Figure PCTCN2016087368-appb-000101
Figure PCTCN2016087368-appb-000101
按照制备化合物4-7的操作方法，以7-丁基-2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪 唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(3-10)(524mg，1.07mmol)为原料去保护基得到白色粉末240mg，产率为58％(二氯甲烷：甲醇＝50：1)。According to the procedure for the preparation of compound 4-7, 7-butyl-2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d] Zin-2-yl)-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (3-10) (524 mg, 1.07 mmol) as a starting material deprotecting group A white powder of 240 mg was obtained in a yield of 58% (dichloromethane:methanol = 50:1).
1H NMR(300MHz,Chloroform-d)δ10.40(brs,1H),7.62(dd,J＝9.0,2.5Hz,1H),7.58(dd,J＝9.6,2.3Hz,1H),7.38(s,1H),4.14–4.03(m,1H),4.01(s,3H),3.44–3.30(m,1H),3.10–2.95(m,1H),2.82–2.62(m,1H),1.84–1.65(m,2H),1.61–1.43(m,1H),1.46–1.33(m,2H),0.94(t,J＝7.1Hz,3H)；MS(ESI)388[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 10.40 (brs, 1H), 7.62 (dd, J = 9.0, 2.5 Hz, 1H), 7.58 (dd, J = 9.6, 2.3 Hz, 1H), 7.38 (s) , 1H), 4.14–4.03 (m, 1H), 4.01 (s, 3H), 3.44–3.30 (m, 1H), 3.10–2.95 (m, 1H), 2.82–2.62 (m, 1H), 1.84–1.65 (m, 2H), 1.61 - 1.43 (m, 1H), 1.46 - 1.33 (m, 2H), 0.94 (t, J = 7.1 Hz, 3H); MS (ESI) 388 [M+H] + .
步骤7. 2-(7-丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺(I-20)：Step 7. 2-(7-Butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H-benzo[d]imidazole- 4-carboxamide (I-20):
Figure PCTCN2016087368-appb-000102
Figure PCTCN2016087368-appb-000102
按照制备化合物I-1的操作方法，以6-氟-2-(7-丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(4-10)(240mg，0.62mmol)为原料氨解得到淡黄色粉末178mg，产率为77％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 6-fluoro-2-(7-butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H -Benzyl[d]imidazole-4-carboxylic acid methyl ester (4-10) (240 mg, 0.62 mmol) was obtained as a crude material (yield: 178 mg,yield: 77% (dichloromethane:methanol = 10:1) .
1H NMR(500MHz,DMSO-d6)δ9.03(brs,1H),7.93(brs,1H),7.67(s,1H),7.55(dd,J＝14.8,2.4,1H),7.53(dd,J＝12.8,2.4,1H),7.52(d,J＝2.4Hz,1H),3.98-3.89(m,1H),3.15(dt,J＝12.4,4.4Hz,1H),2.83(ddd,J＝13.1,8.8,5.0Hz,1H),2.67-2.54(m,2H),1.82–1.71(m,1H),1.69-1.60(m,1H),1.52-1.41(m,2H),1.41–1.27(m,2H),0.90(t,J＝7.3Hz,3H)；13C NMR(125MHz,DMSO-d6)δ165.57,158.66(d,J＝237.0Hz,C-F),149.55,144.43,136.38,129.17,128.78,122.97,110.24(d,J＝26.2Hz,CH-CF),102.18(d,J＝27.9Hz,CH-CF),54.61,42.05,37.09,28.12,26.55,22.66,14.45；HRMS(ESI):m/z Calcd.For C19H22FN4OS[M+H]+:373.1498；Found:373.1490。 1 H NMR (500 MHz, DMSO-d 6 ) δ 9.03 (brs, 1H), 7.93 (brs, 1H), 7.67 (s, 1H), 7.55 (dd, J = 14.8, 2.4, 1H), 7. , J = 12.8, 2.4, 1H), 7.52 (d, J = 2.4 Hz, 1H), 3.98-3.89 (m, 1H), 3.15 (dt, J = 12.4, 4.4 Hz, 1H), 2.83 (ddd, J =13.1, 8.8, 5.0 Hz, 1H), 2.67-2.54 (m, 2H), 1.82–1.71 (m, 1H), 1.69-1.60 (m, 1H), 1.52-1.41 (m, 2H), 1.41–1.27 (m, 2H), 0.90 (t, J = 7.3 Hz, 3H); 13 C NMR (125MHz, DMSO-d 6 ) δ 165.57, 158.66 (d, J = 237.0 Hz, CF), 149.55, 144.43, 136.38, 129. , 128.78, 122.97, 110.24 (d, J = 26.2 Hz, CH-CF), 102.18 (d, J = 27.9 Hz, CH-CF), 54.61, 42.05, 37.09, 28.12, 26.55, 22.66, 14.45; HRMS (ESI) ): m/z Calcd. For C 19 H 22 FN 4 OS [M+H] + : 373.1498; Found: 373.1490.
实施例21. 6-氟-2-(7-异丙基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-21的制备：Example 21. 6-Fluoro-2-(7-isopropyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d] Preparation of imidazole-4-carboxamide, I-21:
步骤1. N-(2-(噻吩-3-基)乙基)异丁酰胺：Step 1. N-(2-(Thien-3-yl)ethyl)isobutyramide:
Figure PCTCN2016087368-appb-000103
Figure PCTCN2016087368-appb-000103
按照制备16-1的操作方法，以2-(噻吩-3-基)乙基胺盐酸盐(492mg，3.0mmol)，异丁酰氯(383mg，3.6mmol)为原料制得浅黄色油状物585mg，产率为99％，直接进入下一步反应。Depending on the procedure of Preparation 16-1, 2-(thien-3-yl)ethylamine hydrochloride (492 mg, 3.0 mmol), isobutyryl chloride (383 mg, 3.6 mmol) The yield was 99% and went directly to the next reaction.
1H NMR(300MHz,Chloroform-d)δ7.29(dd,J＝4.9,3.0Hz,1H),7.01–6.98(m,1H),6.96(dd,J＝4.9,1.3Hz,1H),5.44(brs,1H),3.55–3.47(m,2H),2.85(t,J＝6.9Hz,2 H),2.29(hept,J＝6.8Hz,1H),1.11(d,J＝6.9Hz,6H)；MS(ESI)198[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ7.29 (dd, J = 4.9,3.0Hz, 1H), 7.01-6.98 (m, 1H), 6.96 (dd, J = 4.9,1.3Hz, 1H), 5.44 (brs, 1H), 3.55–3.47 (m, 2H), 2.85 (t, J = 6.9 Hz, 2 H), 2.29 (hept, J = 6.8 Hz, 1H), 1.11 (d, J = 6.9 Hz, 6H) MS (ESI) 198 [M+H] + .
步骤2. 7-异丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 2. 7-Isopropyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000104
Figure PCTCN2016087368-appb-000104
按照制备14-3的操作方法，以N-(2-(噻吩-3-基)乙基)异丁酰胺(16-5)(585mg，3.0mmol)为原料制得白色固体611mg，三步总产率为72％(石油醚：乙酸乙酯＝30：1)。According to the procedure of Preparation 14-3, N-(2-(thien-3-yl)ethyl)isobutyramide (16-5) (585 mg, 3.0 mmol) was used as a starting material to obtain 611 mg of white solid, three-step total The yield was 72% (petroleum ether: ethyl acetate = 30:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ7.14(d,J＝5.0Hz,1H),6.79(d,J＝5.0Hz,1H),5.07–4.97(m,1H),4.28–4.13(m,1H),3.21–2.95(m,1H),2.82–2.64(m,2H),2.06(dt,J＝13.3,6.7Hz,1H),1.47(s,9H),1.14(d,J＝6.7Hz,3H),0.97(d,J＝6.9Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ7.14(d,J＝5.0Hz,1H),6.79(d,J＝5.0Hz,1H),4.94–4.85(m,1H),4.50–4.36(m,1H),3.21–2.95(m,1H),2.82–2.64(m,2H),2.06(dt,J＝13.3,6.7Hz,1H),1.47(s,9H),1.14(d,J＝6.7Hz,3H),0.97(d,J＝6.9Hz,3H)；MS(ESI)304[M+Na]+。Conformation isomer 1 : 1 H NMR (300 MHz, Chloroform-d) δ 7.14 (d, J = 5.0 Hz, 1H), 6.79 (d, J = 5.0 Hz, 1H), 5.07 - 4.97 (m, 1H) , 4.28–4.13 (m, 1H), 3.21–2.95 (m, 1H), 2.82–2.64 (m, 2H), 2.06 (dt, J = 13.3, 6.7 Hz, 1H), 1.47 (s, 9H), 1.14 (d, J = 6.7 Hz, 3H), 0.97 (d, J = 6.9 Hz, 3H); conformational isomer 2: 1 H NMR (300 MHz, Chloroform-d) δ 7.14 (d, J = 5.0 Hz, 1H), 6.79 (d, J = 5.0 Hz, 1H), 4.94 - 4.85 (m, 1H), 4.50 - 4.36 (m, 1H), 3.21 - 2.95 (m, 1H), 2.82 - 2.64 (m, 2H) , 2.06 (dt, J = 13.3, 6.7 Hz, 1H), 1.47 (s, 9H), 1.14 (d, J = 6.7 Hz, 3H), 0.97 (d, J = 6.9 Hz, 3H); MS (ESI) 304 [M+Na] + .
步骤3. 2-溴-7-异丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 3. 2-Bromo-7-isopropyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000105
Figure PCTCN2016087368-appb-000105
按照制备12-3的操作方式，以7-异丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(14-7)(611mg，2.2mmol)为原料经溴代制得672mg白色固体，产率为85％(石油醚：乙酸乙酯＝30：1)。According to the procedure of Preparation 12-3, 4-isopropyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (14-7) (611 mg) </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> <RTIgt;
构象异构体1：1H NMR(300MHz,Chloroform-d)δ6.74(s,1H),4.96–4.84(m,1H),4.50–4.31(m,1H),2.79–2.57(m,2H),2.10–1.89(m,2H),1.46(s,9H),1.11(d,J＝6.7Hz,3H),0.96(d,J＝6.9Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ6.74(s,1H),4.81–4.71(m,1H),4.24–4.05(m,1H),3.26–2.90(m,2H),2.10–1.89(m,2H),1.46(s,9H),1.11(d,J＝6.7Hz,3H),0.96(d,J＝6.9Hz,3H)；MS(ESI)382[M+Na]+。Conformation isomer 1 : 1 H NMR (300 MHz, Chloroform-d) δ 6.74 (s, 1H), 4.96 - 4.84 (m, 1H), 4.50 - 4.31 (m, 1H), 2.79 - 2.57 (m, 2H) ), 2.10–1.89 (m, 2H), 1.46 (s, 9H), 1.11 (d, J = 6.7 Hz, 3H), 0.96 (d, J = 6.9 Hz, 3H); conformation 2: 1 H NMR (300MHz, Chloroform-d) δ 6.74 (s, 1H), 4.81 - 4.71 (m, 1H), 4.24 - 4.05 (m, 1H), 3.26 - 2.90 (m, 2H), 2.10 - 1.89 (m, 2H), 1.46 (s, 9H), 1.11 (d, J = 6.7 Hz, 3H), 0.96 (d, J = 6.9 Hz, 3H); MS (ESI) 382 [M+Na] + .
步骤4. 2-甲酰基-7-异丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 4. 2-Formyl-7-isopropyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000106
Figure PCTCN2016087368-appb-000106
按照制备化合物2-3的操作方法，以2-溴-7-异丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(12-7)(672mg，1.9mmol)为原料制备得到无色油状物240mg， 产率为40％(石油醚：乙酸乙酯＝8：1)。According to the procedure for the preparation of compound 2-3, 2-bromo-7-isopropyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (12 -7) (672 mg, 1.9 mmol) was prepared as a starting material to give 240 mg of colorless oil. The yield was 40% (petroleum ether: ethyl acetate = 8:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ9.84(s,1H),7.47(s,1H),5.15–5.07(m,1H),4.31–4.20(m,1H),3.19–2.94(m,2H),2.17–1.90(m,2H),1.48(s,9H),1.16(d,J＝6.7Hz,3H),0.98(d,J＝6.9Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ9.84(s,1H),7.47(s,1H),4.99–4.91(m,1H),4.54–4.43(m,1H),2.87–2.58(m,2H),2.17–1.90(m,2H),1.48(s,9H),1.16(d,J＝6.7Hz,4H),1.16(d,J＝6.7Hz,3H),0.98(d,J＝6.9Hz,3H)；MS(ESI)332[M+Na]+。Conformation isomer 1 : 1 H NMR (300 MHz, Chloroform-d) δ 9.84 (s, 1H), 7.47 (s, 1H), 5.15 - 5.07 (m, 1H), 4.31 - 4.20 (m, 1H), 3.19–2.94 (m, 2H), 2.17–1.90 (m, 2H), 1.48 (s, 9H), 1.16 (d, J = 6.7 Hz, 3H), 0.98 (d, J = 6.9 Hz, 3H); Isomer 2: 1 H NMR (300 MHz, Chloroform-d) δ 9.84 (s, 1H), 7.47 (s, 1H), 4.99 - 4.91 (m, 1H), 4.54 - 4.43 (m, 1H), 2.87 – 2.58 (m, 2H), 2.17–1.90 (m, 2H), 1.48 (s, 9H), 1.16 (d, J = 6.7 Hz, 4H), 1.16 (d, J = 6.7 Hz, 3H), 0.98 ( d, J = 6.9 Hz, 3H); MS (ESI) 332 [M+Na] + .
步骤5. 2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-7-异丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 5. 2-(6-Fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-7-isopropyl-4,7-dihydrothieno[2, 3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000107
Figure PCTCN2016087368-appb-000107
按照制备化合物3-7的操作方法，以2-甲酰基-7-异丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(2-7)(240mg，0.78mmol)和2,3-二氨基-5-氟苯甲酸甲酯(143mg，0.78mmol)为原料制备得到黄色油状物333mg，产率为90％(石油醚：乙酸乙酯＝3：1)。According to the procedure for the preparation of compound 3-7, 2-formyl-7-isopropyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester ( 2-7) (240 mg, 0.78 mmol) and 2,3-diamino-5-fluorobenzoic acid methyl ester (143 mg, 0.78 mmol) were used as starting materials to give 333 mg of yellow oil (yield: 90%) Ethyl ester = 3:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ10.51(s,1H),7.74–7.47(m,2H),7.38(s,1H),5.15–5.04(m,1H),4.33–4.21(m,1H),4.00(s,3H),3.21–2.99(m,1H),2.89–2.54(m,2H),2.25–2.06(m,1H),1.18(d,J＝6.6Hz,3H),1.02(d,J＝6.8Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ10.51(s,1H),7.74–7.47(m,2H),7.38(s,1H),5.00–4.90(m,1H),4.57–4.41(m,1H),4.00(s,3H),3.21–2.99(m,1H),2.89–2.54(m,2H),2.25–2.06(m,1H),1.18(d,J＝6.6Hz,3H),1.02(d,J＝6.8Hz,3H)；MS(ESI)475[M+H]+。Conformation isomer 1 : 1 H NMR (300 MHz, Chloroform-d) δ 10.51 (s, 1H), 7.74 - 7.47 (m, 2H), 7.38 (s, 1H), 5.15 - 5.04 (m, 1H), 4.33–4.21 (m, 1H), 4.00 (s, 3H), 3.21–2.99 (m, 1H), 2.89–2.54 (m, 2H), 2.25–2.06 (m, 1H), 1.18 (d, J=6.6) Hz, 3H), 1.02 (d, J = 6.8 Hz, 3H); conformational isomer 2: 1 H NMR (300 MHz, Chloroform-d) δ 10.51 (s, 1H), 7.74 - 7.47 (m, 2H) , 7.38 (s, 1H), 5.00–4.90 (m, 1H), 4.57–4.41 (m, 1H), 4.00 (s, 3H), 3.21–2.99 (m, 1H), 2.89–2.54 (m, 2H) , 2.25 - 2.06 (m, 1H), 1.18 (d, J = 6.6 Hz, 3H), 1.02 (d, J = 6.8 Hz, 3H); MS (ESI) 475 [M+H] + .
步骤6. 6-氟-2-(7-异丙基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 6. 6-Fluoro-2-(7-isopropyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole -4-methyl formate:
Figure PCTCN2016087368-appb-000108
Figure PCTCN2016087368-appb-000108
按照制备化合物4-7的操作方法，以2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-7-异丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(3-11)(333mg，0.89mmol)为原料去保护基得到淡黄色粉末248mg，产率为75％(二氯甲烷：甲醇＝50：1)。According to the procedure for the preparation of compound 4-7, 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-7-isopropyl-4,7 -Dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (3-11) (333 mg, 0.89 mmol) as a starting material deprotecting group afforded 248 mg of pale yellow powder, yield 75 % (dichloromethane: methanol = 50:1).
1H NMR(300MHz,Chloroform-d)δ10.42(brs,1H),7.62(dd,J＝14.0,2.3Hz,1H), 7.59(dd,J＝14.5,2.3Hz,1H),7.39(s,1H),4.02(s,3H),3.39(ddd,J＝13.0,5.3,2.7Hz,1H),3.06–2.90(m,1H),2.81–2.58(m,2H),2.25–2.10(m,1H),1.16(d,J＝7.0Hz,3H),0.95(d,J＝6.9Hz,3H)；MS(ESI)374[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ10.42 (brs, 1H), 7.62 (dd, J = 14.0,2.3Hz, 1H), 7.59 (dd, J = 14.5,2.3Hz, 1H), 7.39 (s , 1H), 4.02 (s, 3H), 3.39 (ddd, J = 13.0, 5.3, 2.7 Hz, 1H), 3.06 - 2.90 (m, 1H), 2.81 - 2.58 (m, 2H), 2.25 - 2.10 (m , 1H), 1.16 (d, J = 7.0 Hz, 3H), 0.95 (d, J = 6.9 Hz, 3H); MS (ESI) 374 [M+H] + .
步骤7. 6-氟-2-(7-异丙基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-21)：Step 7. 6-Fluoro-2-(7-isopropyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole 4-carboxamide (I-21):
Figure PCTCN2016087368-appb-000109
Figure PCTCN2016087368-appb-000109
按照制备化合物I-1的操作方法，以6-氟-2-(7-异丙基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(4-11)(248mg，0.66mmol)为原料氨解得到淡棕色粉末166mg，产率为70％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 6-fluoro-2-(7-isopropyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)- 1H-benzo[d]imidazole-4-carboxylic acid methyl ester (4-11) (248 mg, 0.66 mmol) was obtained by aminolysis to give 166 mg of pale brown powder, yield 70% (dichloromethane:methanol = 10:1) ).
1H NMR(500MHz,DMSO-d6)δ9.08(brs,1H),7.97(brs,1H),7.76(s,1H),7.55(t,J＝10.8Hz,2H),4.24–4.17(m,1H),3.35–3.28(m,1H),3.03–2.95(m,1H),2.85-2.75(m,1H),2.74–2.65(m,1H),2.23-2.15(m,1H),1.11(d,J＝6.9Hz,3H),0.95(d,J＝6.9Hz,3H)；13C NMR(125MHz,DMSO-d6)δ165.33,158.77(d,J＝236.9Hz),149.04,138.46,136.85,136.09,129.99,129.05,123.34,110.44(d,J＝24.4Hz),101.82(d,J＝26.9Hz),59.80,42.35,33.49,29.49,19.62,17.32.HRMS(ESI):m/z Calcd.For C18H20FN4OS[M+H]+:359.1342；Found:359.1336。 1 H NMR (500MHz, DMSO- d 6) δ9.08 (brs, 1H), 7.97 (brs, 1H), 7.76 (s, 1H), 7.55 (t, J = 10.8Hz, 2H), 4.24-4.17 ( m,1H), 3.35–3.28 (m,1H), 3.03–2.95 (m,1H), 2.85-2.75 (m,1H), 2.74–2.65 (m,1H), 2.23-2.15 (m,1H), 1.11 (d, J = 6.9 Hz, 3H), 0.95 (d, J = 6.9 Hz, 3H); 13 C NMR (125 MHz, DMSO-d 6 ) δ 165.33, 158.77 (d, J = 236.9 Hz), 149.04, 138.46 , 136.85, 136.09, 129.99, 129.05, 123.34, 110.44 (d, J = 24.4 Hz), 101.82 (d, J = 26.9 Hz), 59.80, 42.35, 33.49, 29.49, 19.62, 17.32. HRMS (ESI): m/ z Calcd. For C 18 H 20 FN 4 OS [M+H] + :359.1342; Found: 359.1336.
实施例22. 2-(7-环丙基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺，I-22的制备：Example 22. 2-(7-Cyclopropyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H-benzo[d] Preparation of imidazole-4-carboxamide, I-22:
步骤1. N-(2-(噻吩-3-基)乙基)环丙基甲酰胺：Step 1. N-(2-(Thien-3-yl)ethyl)cyclopropyl formamide:
Figure PCTCN2016087368-appb-000110
Figure PCTCN2016087368-appb-000110
按照制备16-1的操作方法，以2-(噻吩-3-基)乙基胺盐酸盐(492mg，3.0mmol)，环丙基甲酰氯(376mg，3.6mmol)为原料制得浅黄色油状物579mg，产率为99％，直接进入下一步反应。According to the procedure of Preparation 16-1, 2-(thiophen-3-yl)ethylamine hydrochloride (492 mg, 3.0 mmol), cyclopropylcarbonyl chloride (376 mg, 3.6 mmol) The product was 579 mg in a yield of 99% and directly proceeded to the next reaction.
1H NMR(300MHz,Chloroform-d)δ7.32–7.27(m,1H),7.02(s,1H),6.96(d,J＝4.8Hz,1H),5.64(brs,0H),3.54(dd,J＝6.7,6.5Hz,2H),2.86(t,J＝6.7Hz,2H),1.32–1.21(m,1H),0.99–0.92(m,2H),0.75–0.66(m,2H)；MS(ESI)218[M+Na]+。 1 H NMR (300 MHz, Chloroform-d) δ 7.32 - 7.27 (m, 1H), 7.02 (s, 1H), 6.96 (d, J = 4.8 Hz, 1H), 5.64 (brs, 0H), 3.54 (dd , J = 6.7, 6.5 Hz, 2H), 2.86 (t, J = 6.7 Hz, 2H), 1.32 - 1.21 (m, 1H), 0.99 - 0.92 (m, 2H), 0.75 - 0.66 (m, 2H); MS (ESI) 218 [M+Na] + .
步骤2. 7-环丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯： Step 2. 7-Cyclopropyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000111
Figure PCTCN2016087368-appb-000111
按照制备14-3的操作方法，以N-(2-(噻吩-3-基)乙基)环丙基甲酰胺(16-6)(579mg，3.0mmol)为原料制得白色固体373mg，三步总产率为45％(石油醚：乙酸乙酯＝30：1)。According to the procedure of Preparation 14-3, N-(2-(thiophen-3-yl)ethyl)cyclopropylcarboxamide (16-6) (579 mg, 3.0 mmol) The total yield in steps was 45% (petroleum ether: ethyl acetate = 30:1).
1H NMR(300MHz,Chloroform-d)δ7.13(d,J＝5.1Hz,1H),6.77(d,J＝5.1Hz,1H),4.62–4.25(m,1H),3.26–3.06(m,1H),2.81–2.52(m,2H),1.47(d,J＝0.7Hz,9H),1.32–1.18(m,1H),0.83–0.61(m,2H),0.59–0.47(m,2H)；MS(ESI)302[M+Na]+。 1 H NMR (300 MHz, Chloroform-d) δ 7.13 (d, J = 5.1 Hz, 1H), 6.77 (d, J = 5.1 Hz, 1H), 4.62 - 4.25 (m, 1H), 3.26 - 3.06 (m) , 1H), 2.81–2.52 (m, 2H), 1.47 (d, J = 0.7 Hz, 9H), 1.32–1.18 (m, 1H), 0.83–0.61 (m, 2H), 0.59–0.47 (m, 2H) ); MS (ESI) 302 [M+Na] + .
步骤3. 2-溴-7-环丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 3. 2-Bromo-7-cyclopropyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000112
Figure PCTCN2016087368-appb-000112
按照制备12-3的操作方式，以7-环丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(14-8)(373mg，1.4mmol)为原料经溴代制得382mg无色油状物，产率为79％(石油醚：乙酸乙酯＝30：1)。According to the procedure of Preparation 12-3, 7-cyclopropyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (14-8) (373 mg) </RTI> </RTI> </RTI> <RTI ID=0.0></RTI> </RTI> </RTI>
1H NMR(300MHz,Chloroform-d)δ6.73(s,1H),4.42–4.29(m,2H),3.22–3.07(m,1H),2.73–2.58(m,1H),2.57–2.44(m,1H),1.46(s,9H),1.31–1.17(m,1H),0.80–0.60(m,2H),0.61–0.38(m,2H)；MS(ESI)380[M+Na]+。 1 H NMR (300 MHz, Chloroform-d) δ 6.73 (s, 1H), 4.42 - 4.29 (m, 2H), 3.22 - 3.07 (m, 1H), 2.73 - 2.58 (m, 1H), 2.57 - 2.44 ( m,1H), 1.46 (s, 9H), 1.31 - 1.17 (m, 1H), 0.80 - 0.60 (m, 2H), 0.61 - 0.38 (m, 2H); MS (ESI) 380 [M+Na] + .
步骤4. 7-环丙基-2-甲酰基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 4. 7-Cyclopropyl-2-formyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000113
Figure PCTCN2016087368-appb-000113
按照制备化合物2-3的操作方法，以2-溴-7-环丙基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(12-8)(382mg，1.1mmol)为原料制备得到淡黄色油状物302mg，产率为89％(石油醚：乙酸乙酯＝8：1)。According to the procedure for the preparation of compound 2-3, 2-bromo-7-cyclopropyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (12 -8) (382 mg, 1.1 mmol) was obtained as a crude material (yield: EtOAc: EtOAc: EtOAc:
1H NMR(300MHz,Chloroform-d)δ9.84(s,1H),7.45(s,1H),4.53(brs,1H),4.40(brs,1H),3.27–3.00(m,1H),2.87–2.58(m,2H),1.67–1.57(m,1H),1.47(s,9H),1.34–1.15(m,1H),0.91–0.64(m,2H),0.65–0.49(m,2H)；MS(ESI)330[M+Na]+。 1 H NMR (300 MHz, Chloroform-d) δ 9.84 (s, 1H), 7.45 (s, 1H), 4.53 (brs, 1H), 4.40 (brs, 1H), 3.27 - 3.00 (m, 1H), 2.87 –2.58 (m, 2H), 1.67–1.57 (m, 1H), 1.47 (s, 9H), 1.34–1.15 (m, 1H), 0.91–0.64 (m, 2H), 0.65–0.49 (m, 2H) ;MS (ESI) 330 [M+Na] + .
步骤5. 7-环丙基-2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯： Step 5. 7-Cyclopropyl-2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7-dihydrothieno[2, 3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000114
Figure PCTCN2016087368-appb-000114
按照制备化合物3-7的操作方法，以7-环丙基-2-甲酰基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(2-8)(302mg，0.98mmol)和2,3-二氨基-5-氟苯甲酸甲酯(181mg，0.98mmol)为原料制备得到白色固体438mg，产率为95％(石油醚：乙酸乙酯＝3：1)。According to the procedure for the preparation of compound 3-7, 7-cyclopropyl-2-formyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester ( 2-8) (302 mg, 0.98 mmol) and 2,3-diamino-5-fluorobenzoic acid methyl ester (181 mg, 0.98 mmol) were used as starting materials to give 438 mg of white solid (yield: 95%) Ester = 3:1).
1H NMR(300MHz,Chloroform-d)δ10.42(brs,1H),7.63(dd,J＝8.9,2.4Hz,1H),7.58(dd,J＝9.6,2.4Hz,1H),7.36(s,1H),4.53(brs,1H),4.43(brs,2H),4.01(s,3H),3.29–3.10(m,1H),2.86–2.58(m,2H),1.49(s,9H),0.92–0.67(m,2H),0.65–0.53(m,2H)；MS(ESI)472[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 10.42 (brs, 1H), 7.63 (dd, J = 8.9, 2.4 Hz, 1H), 7.58 (dd, J = 9.6, 2.4 Hz, 1H), 7.36 (s) , 1H), 4.53 (brs, 1H), 4.43 (brs, 2H), 4.01 (s, 3H), 3.29 - 3.10 (m, 1H), 2.86 - 2.58 (m, 2H), 1.49 (s, 9H), 0.92 - 0.67 (m, 2H), 0.65 - 0.53 (m, 2H); MS (ESI) 472 [M+H] + .
步骤6. 2-(7-环丙基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯：Step 6. 2-(7-Cyclopropyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H-benzo[d]imidazole -4-methyl formate:
Figure PCTCN2016087368-appb-000115
Figure PCTCN2016087368-appb-000115
按照制备化合物4-7的操作方法，以7-环丙基-2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(3-12)(438mg，0.80mmol)为原料去保护基得到白色粉末298mg，产率为86％(二氯甲烷：甲醇＝50：1)。According to the procedure for the preparation of compound 4-7, 7-cyclopropyl-2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7 -Dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (3-12) (438 mg, 0.80 mmol) as a starting material deprotecting group afforded 298 mg of white powder, yield 86% (Dichloromethane: methanol = 50:1).
1H NMR(300MHz,Chloroform-d)δ10.41(s,1H),7.63(dd,J＝8.9,2.3Hz,1H),7.57(d,J＝9.7Hz,1H),7.38(s,1H),4.01(s,3H),3.38(ddd,J＝12.4,5.6,2.7Hz,1H),3.14(dd,J＝9.5,2.2Hz,1H),2.99(ddd,J＝12.3,10.0,4.7Hz,1H),2.87–2.72(m,1H),2.71–2.60(m,1H),1.20–1.05(m,1H),0.84–0.61(m,2H),0.60–0.41(m,2H)；MS(ESI)372[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 10.41 (s, 1H), 7.63 (dd, J = 8.9, 2.3 Hz, 1H), 7.57 (d, J = 9.7 Hz, 1H), 7.38 (s, 1H) ), 4.01 (s, 3H), 3.38 (ddd, J = 12.4, 5.6, 2.7 Hz, 1H), 3.14 (dd, J = 9.5, 2.2 Hz, 1H), 2.99 (ddd, J = 12.3, 10.0, 4.7) Hz, 1H), 2.87 - 2.72 (m, 1H), 2.71 - 2.60 (m, 1H), 1.20 - 1.05 (m, 1H), 0.84 - 0.61 (m, 2H), 0.60 - 0.41 (m, 2H); MS (ESI) 372 [M + H] + .
步骤7. 2-(7-环丙基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺(I-22)：Step 7. 2-(7-Cyclopropyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H-benzo[d]imidazole 4-carboxamide (I-22):
Figure PCTCN2016087368-appb-000116
Figure PCTCN2016087368-appb-000116
按照制备化合物I-1的操作方法，以2-(7-环丙基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯(4-12)(298mg，0.60mmol)为原料氨解得到淡 棕色粉末260mg，产率为91％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 2-(7-cyclopropyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro- 1H-benzo[d]imidazole-4-carboxylic acid methyl ester (4-12) (298 mg, 0.60 mmol) was obtained as a raw material. The brown powder was 260 mg in a yield of 91% (dichloromethane:methanol = 10:1).
1H NMR(300MHz,DMSO-d6)δ9.00(brs,1H),7.88(brs,1H),7.68(s,1H),7.53(dd,J＝11.0,2.6Hz,1H),7.49(dd,J＝9.6,2.6Hz,1H),3.28-3.17(m,2H),2.92–2.76(m,1H),2.76–2.54(m,2H),1.18–0.98(m,2H),0.75-0.62(m,1H),0.6–0.40(m,2H)；13C NMR(125MHz,DMSO-d6)δ165.56,158.67(d,J＝236.6Hz,C-F),149.54,143.14,135.92,129.37,128.80,122.83,110.24(d,J＝26.2Hz,CH-CF),102.16(d,J＝24.6Hz,CH-CF),60.43,42.54,29.50,25.92,18.06,4.30,3.84；HRMS(ESI):m/z Calcd.For C18H18FN4OS[M+H]+:357.1185；Found:357.1179。 1 H NMR (300MHz, DMSO- d 6) δ9.00 (brs, 1H), 7.88 (brs, 1H), 7.68 (s, 1H), 7.53 (dd, J = 11.0,2.6Hz, 1H), 7.49 ( Dd, J=9.6, 2.6 Hz, 1H), 3.28-3.17 (m, 2H), 2.92–2.76 (m, 1H), 2.76–2.54 (m, 2H), 1.18–0.98 (m, 2H), 0.75- 0.62 (m, 1H), 0.6 - 0.40 (m, 2H); 13 C NMR (125MHz, DMSO-d 6 ) δ 165.56, 158.67 (d, J = 236.6 Hz, CF), 149.54, 143.14, 135.92, 129.37, 128.80 , 122.83, 110.24 (d, J = 26.2 Hz, CH-CF), 102.16 (d, J = 24.6 Hz, CH-CF), 60.43, 42.54, 29.50, 25.92, 18.06, 4.30, 3.84; HRMS (ESI): m/z Calcd. For C 18 H 18 FN 4 OS [M+H] + : 357.1185; Found: 357.1179.
实施例23. 2-(7-叔丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺，I-23的制备：Example 23. 2-(7-tert-Butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H-benzo[d] Preparation of imidazole-4-carboxamide, I-23:
步骤1. N-(2-(噻吩-3-基)乙基)特戊酰胺：Step 1. N-(2-(Thien-3-yl)ethyl)pivalamide:
Figure PCTCN2016087368-appb-000117
Figure PCTCN2016087368-appb-000117
按照制备16-1的操作方法，以2-(噻吩-3-基)乙基胺盐酸盐(492mg，3.0mmol)，特戊酰氯(434mg，3.6mmol)为原料制得浅黄色油状物627mg，产率为99％，直接进入下一步反应。According to the procedure of Preparation 16-1, 2-(thiophen-3-yl)ethylamine hydrochloride (492 mg, 3.0 mmol), pivaloyl chloride (434 mg, 3.6 mmol) The yield was 99% and went directly to the next reaction.
1H NMR(300MHz,Chloroform-d)δ7.31–7.27(m,1H),7.01–6.93(m,2H),5.6(brs,1H),3.53–3,45(m,,2H),2.85(t,J＝6.8Hz,2H),1.14(s,9H)；MS(ESI)212[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 7.31 - 7.27 (m, 1H), 7.01 - 6.93 (m, 2H), 5.6 (brs, 1H), 3.53 - 3, 45 (m,, 2H), 2.85 (t, J = 6.8 Hz, 2H), 1.14 (s, 9H); MS (ESI) 212 [M+H] + .
步骤2. 7-叔丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 2. 7-tert-Butyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000118
Figure PCTCN2016087368-appb-000118
按照制备14-3的操作方法，以N-(2-(噻吩-3-基)乙基)环丙基甲酰胺(16-7)(627mg，3.0mmol)为原料制得白色固体697mg，三步总产率为79％(石油醚：乙酸乙酯＝30：1)。According to the procedure of Preparation 14-3, N-(2-(thiophen-3-yl)ethyl)cyclopropylformamide (16-7) (627 mg, 3.0 mmol) The total yield of the step was 79% (petroleum ether: ethyl acetate = 30:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ7.13(dd,J＝7.3,5.1Hz,1H),6.79(dd,J＝9.4,5.1Hz,1H),5.14(s,1H),4.24(dd,J＝13.8,5.7Hz,1H),3.32–3.09(m,1H),2.82–2.48(m,2H),1.46(s,9H),1.08(s,6H),1.07(s,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ7.13(dd,J＝7.3,5.1Hz,1H),6.79(dd,J＝9.4,5.1Hz,1H),4.96(s,1H),4.47(dd,J＝13.6,6.1Hz,1H),3.32–3.09(m,1H),2.82–2.48(m,2H),1.46(s,9H),1.08(s,6H),1.07(s,3H)；MS(ESI)318[M+Na]+。Conformation isomer 1 : 1 H NMR (300 MHz, Chloroform-d) δ 7.13 (dd, J = 7.3, 5.1 Hz, 1H), 6.79 (dd, J = 9.4, 5.1 Hz, 1H), 5.14 (s, 1H), 4.24 (dd, J = 13.8, 5.7 Hz, 1H), 3.32 - 3.09 (m, 1H), 2.82 - 2.48 (m, 2H), 1.46 (s, 9H), 1.08 (s, 6H), 1.07 (s, 3H); conformational isomer 2: 1 H NMR (300 MHz, Chloroform-d) δ 7.13 (dd, J = 7.3, 5.1 Hz, 1H), 6.79 (dd, J = 9.4, 5.1 Hz, 1H) ), 4.96 (s, 1H), 4.47 (dd, J = 13.6, 6.1 Hz, 1H), 3.32 - 3.09 (m, 1H), 2.82 - 2.48 (m, 2H), 1.46 (s, 9H), 1.08 ( s, 6H), 1.07 (s, 3H); MS (ESI) 318 [M+Na] + .
步骤3. 2-溴-7-叔丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯： Step 3. 2-Bromo-7-tert-butyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000119
Figure PCTCN2016087368-appb-000119
按照制备12-3的操作方式，以7-叔丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(14-9)(697mg，2.4mmol)为原料经溴代制得694mg无色油状物，产率为77％(石油醚：乙酸乙酯＝30：1)。According to the procedure of Preparation 12-3, tert-butyl 7-tert-butyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylate (14-9) (697 mg) </ RTI> </ RTI> </ RTI> </RTI> </RTI> </RTI> </ RTI> </ RTI> </ RTI>
构象异构体1：1H NMR(300MHz,Chloroform-d)δ6.77(s,1H),5.02(s,1H),4.42(dd,J＝13.7,6.4Hz,1H),3.33–3.05(m,1H),2.76–2.39(m,2H),1.46(s,9H),1.06(s,9H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ6.74(s,1H),4.83(s,1H),4.20(dd,J＝13.9,6.4Hz,1H),3.33–3.05(m,1H),2.76–2.39(m,2H),1.46(s,9H),1.06(s,9H)；MS(ESI)396[M+Na]+。Conformation isomer 1 : 1 H NMR (300 MHz, Chloroform-d) δ 6.77 (s, 1H), 5.02 (s, 1H), 4.42 (dd, J = 13.7, 6.4 Hz, 1H), 3.33 - 3.05 ( m, 1H), 2.76 - 2.39 (m, 2H), 1.46 (s, 9H), 1.06 (s, 9H); conformation 2: 1 H NMR (300 MHz, Chloroform-d) δ 6.74 (s, 1H), 4.83 (s, 1H), 4.20 (dd, J = 13.9, 6.4 Hz, 1H), 3.33 - 3.05 (m, 1H), 2.76 - 2.39 (m, 2H), 1.46 (s, 9H), 1.06 (s, 9H); MS (ESI) 396 [M+Na] + .
步骤4. 7-叔丁基-2-甲酰基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 4. 7-tert-Butyl-2-formyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000120
Figure PCTCN2016087368-appb-000120
按照制备化合物2-3的操作方法，以2-溴-7-叔丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(12-9)(694mg，1.9mmol)为原料制备得到淡黄色油状物223mg，产率为37％(石油醚：乙酸乙酯＝8：1)。According to the procedure for the preparation of compound 2-3, 2-bromo-7-tert-butyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (12) -9) (694 mg, 1.9 mmol) was obtained as a crude material (yield: 223 mg,yield: 37%) (yield: petroleum ether: ethyl acetate = 8:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ9.85(s,1H),7.49(s,1H),5.20(s,1H),4.50(dd,J＝13.7,6.2Hz,1H),3.37–3.08(m,1H),2.86–2.57(m,2H),1.47(s,9H),1.09(s,9H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ9.84(s,1H),7.47(s,1H),5.00(s,1H),4.29(dd,J＝14.1,5.6Hz,1H),3.37–3.08(m,1H),2.86–2.57(m,2H),1.47(s,9H),1.09(s,9H)；MS(ESI)346[M+Na]+。Conformational isomer 1: 1 H NMR (300 MHz, Chloroform-d) δ 9.85 (s, 1H), 7.49 (s, 1H), 5.20 (s, 1H), 4.50 (dd, J = 13.7, 6.2 Hz, 1H), 3.37–3.08 (m, 1H), 2.86–2.57 (m, 2H), 1.47 (s, 9H), 1.09 (s, 9H); conformation 2: 1 H NMR (300 MHz, Chloroform-d ) δ 9.84 (s, 1H), 7.47 (s, 1H), 5.00 (s, 1H), 4.29 (dd, J = 14.1, 5.6 Hz, 1H), 3.37 - 3.08 (m, 1H), 2.86 - 2.57 (m, 2H), 1.47 (s, 9H), 1.09 (s, 9H); MS (ESI) 346 [M+Na] + .
步骤5. 7-叔丁基-2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 5. 7-tert-Butyl-2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7-dihydrothieno[2, 3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000121
Figure PCTCN2016087368-appb-000121
按照制备化合物3-7的操作方法，以7-叔丁基-2-甲酰基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(2-9)(223mg，0.69mmol)和2,3-二氨基-5-氟苯甲酸甲酯(127mg，0.69mmol)为原料制备得到淡棕色泡沫状固体313mg，产率为93％(石油醚：乙酸乙酯＝3：1)。 According to the procedure for the preparation of compound 3-7, 7-tert-butyl-2-formyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester ( 2-9) (223 mg, 0.69 mmol) and 2,3-diamino-5-fluorobenzoic acid methyl ester (127 mg, 0.69 mmol) were used as starting materials to give 313 mg of pale brown foamy solid (yield: 93%) : ethyl acetate = 3:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ10.42(s,1H),7.64(d,J＝8.9Hz,1H),7.60(d,J＝9.6Hz,1H),7.40(s,1H),5.21(s,1H),4.52(dd,J＝12.2,7.3Hz,1H),4.02(s,3H),3.38–3.09(m,1H),2.88–2.53(m,2H),1.48(s,9H),1.13(s,9H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ10.42(s,1H),7.64(d,J＝8.9Hz,1H),7.60(d,J＝9.6Hz,1H),7.40(s,1H),5.01(s,1H),4.30(dd,J＝13.3,4.7Hz,1H),4.02(s,3H),3.38–3.09(m,1H),2.88–2.53(m,2H),1.48(s,9H),1.13(s,9H)；MS(ESI)488[M+H]+。Conformation isomer 1 : 1 H NMR (300 MHz, Chloroform-d) δ 10.42 (s, 1H), 7.64 (d, J = 8.9 Hz, 1H), 7.60 (d, J = 9.6 Hz, 1H), 7.40 (s, 1H), 5.21 (s, 1H), 4.52 (dd, J = 12.2, 7.3 Hz, 1H), 4.02 (s, 3H), 3.38 - 3.09 (m, 1H), 2.88 - 2.53 (m, 2H) ), 1.48 (s, 9H), 1.13 (s, 9H); conformational isomer 2: 1 H NMR (300 MHz, Chloroform-d) δ 10.42 (s, 1H), 7.64 (d, J = 8.9 Hz, 1H), 7.60 (d, J = 9.6 Hz, 1H), 7.40 (s, 1H), 5.01 (s, 1H), 4.30 (dd, J = 13.3, 4.7 Hz, 1H), 4.02 (s, 3H), 3.38–3.09 (m, 1H), 2.88–2.53 (m, 2H), 1.48 (s, 9H), 1.13 (s, 9H); MS (ESI) 488 [M+H] + .
步骤6. 2-(7-叔丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯：Step 6. 2-(7-tert-Butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H-benzo[d]imidazole -4-methyl formate:
Figure PCTCN2016087368-appb-000122
Figure PCTCN2016087368-appb-000122
按照制备化合物4-7的操作方法，以7-叔丁基-2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(3-13)(313mg，0.64mmol)为原料去保护基得到黄褐色粉末234mg，产率为94％(二氯甲烷：甲醇＝50：1)。According to the procedure for the preparation of compound 4-7, 7-tert-butyl-2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7 -Dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (3-13) (313 mg, 0.64 mmol) as a starting material deprotecting group afforded 234 mg of a tan powder, yield 94 % (dichloromethane: methanol = 50:1).
1H NMR(300MHz,Chloroform-d)δ10.40(s,1H),7.63(dd,J＝15.5,1.8Hz,1H),7.60(dd,J＝15.8,1.7Hz,1H),7.39(s,1H),4.02(s,3H),3.84(s,1H),3.43–3.31(m,1H),2.98–2.85(m,1H),2.80–2.58(m,2H),1.13(s,9H)；MS(ESI)388[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ10.40 (s, 1H), 7.63 (dd, J = 15.5,1.8Hz, 1H), 7.60 (dd, J = 15.8,1.7Hz, 1H), 7.39 (s , 1H), 4.02 (s, 3H), 3.84 (s, 1H), 3.43 - 3.31 (m, 1H), 2.98 - 2.85 (m, 1H), 2.80 - 2.58 (m, 2H), 1.13 (s, 9H) MS (ESI) 388 [M+H] + .
步骤7. 2-(7-叔丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺(I-23)：Step 7. 2-(7-tert-Butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H-benzo[d]imidazole 4-carboxamide (I-23):
Figure PCTCN2016087368-appb-000123
Figure PCTCN2016087368-appb-000123
按照制备化合物I-1的操作方法，以2-(7-叔丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯(4-13)(234mg，0.60mmol)为原料氨解得到黄褐色粉末144mg，产率为65％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 2-(7-tert-butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro- 1H-benzo[d]imidazole-4-carboxylic acid methyl ester (4-13) (234 mg, 0.60 mmol) was used as a starting material to give a yellow brown powder 144 mg (yield: methylene chloride:methanol = 10:1). ).
1H NMR(300MHz,DMSO-d6)δ9.08(brs,1H),7.88(brs,1H),7.67(brs,1H),7.56(dd,J＝8.4,2.5Hz,1H),7.52(dd,J＝8.0,2.5Hz,1H),3.73(s,1H),3.21(t,J＝4.0Hz,1H),2.84–2.70(m,1H),2.66–2.54(m,2H),1.07(s,9H)；13C NMR(125MHz,DMSO-d6)δ165.40,158.71(d,J＝235.0Hz,C-F),149.29,140.61,139.06,138.61,136.02,128.94,123.26,110.34(d,J＝30.3Hz,CH-CF),101.80(d,J＝25.4Hz,CH-CF),64.77,43.02,35.89,27.42,27.35；HRMS(ESI):m/z Calcd.For C19H22FN4OS[M+H]+:373.1498；Found: 373.1490。 1 H NMR (300MHz, DMSO- d 6) δ9.08 (brs, 1H), 7.88 (brs, 1H), 7.67 (brs, 1H), 7.56 (dd, J = 8.4,2.5Hz, 1H), 7.52 ( Dd, J = 8.0, 2.5 Hz, 1H), 3.73 (s, 1H), 3.21 (t, J = 4.0 Hz, 1H), 2.84 - 2.70 (m, 1H), 2.66 - 2.54 (m, 2H), 1.07 (s, 9H); 13 C NMR (125MHz, DMSO-d 6 ) δ 165.40, 158.71 (d, J = 235.0 Hz, CF), 149.29, 140.61, 139.06, 138.61, 136.02, 128.94, 123.26, 110.34 (d, J =30.3 Hz, CH-CF), 101.80 (d, J = 25.4 Hz, CH-CF), 64.77, 43.02, 35.89, 27.42, 27.35; HRMS (ESI): m/z Calcd. For C 19 H 22 FN 4 OS [M+H] + : 373.1498; Found: 373.1490.
实施例24. (S)-2-(7-叔丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺，I-24的制备：Example 24. (S)-2-(7-tert-Butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H-benzene And [d] imidazole-4-carboxamide, preparation of I-24:
步骤1. (S)-2-(7-叔丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯：Step 1. (S)-2-(7-tert-Butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H-benzo [d] Methyl imidazole-4-carboxylate:
Figure PCTCN2016087368-appb-000124
Figure PCTCN2016087368-appb-000124
将2-(7-叔丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯(4-13)经手性制备液相分离得到白色粉末状固体，其绝对构型由单晶X射线衍射确定。分离条件为：色谱柱型号：Chiralpak As；色谱柱规格：0.46cm I.D.×25cm L；进样量：5μL；流动相：CO2/IPA/DEA＝60/40/0.1(v/v/v)；流速：2.5ml/min；检测条件：UVλ＝254nm；柱温：35℃。2-(7-tert-Butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H-benzo[d]imidazole-4 Methyl formate (4-13) was isolated by chiral preparative liquid phase to give a white powdery solid whose absolute configuration was determined by single crystal X-ray diffraction. The separation conditions were: column type: Chiralpak As; column size: 0.46 cm ID × 25 cm L; injection volume: 5 μL; mobile phase: CO 2 /IPA/DEA=60/40/0.1 (v/v/v) Flow rate: 2.5 ml/min; detection conditions: UV λ = 254 nm; column temperature: 35 ° C.
1H NMR(300MHz,Chloroform-d)δ10.40(s,1H),7.63(dd,J＝15.5,1.8Hz,1H),7.60(dd,J＝15.8,1.7Hz,1H),7.39(s,1H),4.02(s,3H),3.84(s,1H),3.43–3.31(m,1H),2.98–2.85(m,1H),2.80–2.58(m,2H),1.13(s,9H)；MS(ESI)388[M+H]+；[α]20D–169(c,0.2,MeOH)；tR＝3.60min。 1 H NMR (300MHz, Chloroform- d) δ10.40 (s, 1H), 7.63 (dd, J = 15.5,1.8Hz, 1H), 7.60 (dd, J = 15.8,1.7Hz, 1H), 7.39 (s , 1H), 4.02 (s, 3H), 3.84 (s, 1H), 3.43 - 3.31 (m, 1H), 2.98 - 2.85 (m, 1H), 2.80 - 2.58 (m, 2H), 1.13 (s, 9H) ); MS (ESI) 388 [ M + H] +; [α] 20D-169 (c, 0.2, MeOH); t R = 3.60min.
步骤2. (S)-2-(7-叔丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺(I-24)：Step 2. (S)-2-(7-tert-Butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H-benzo [d]Imidazole-4-carboxamide (I-24):
Figure PCTCN2016087368-appb-000125
Figure PCTCN2016087368-appb-000125
按照制备化合物I-1的操作方法，以(S)-2-(7-叔丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯(S-4-13)(200mg，0.52mmol)为原料氨解得到白色粉末125mg，产率为65％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, (S)-2-(7-tert-butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)- 6-Fluoro-1H-benzo[d]imidazole-4-carboxylic acid methyl ester (S-4-13) (200 mg, 0.52 mmol) was obtained as a crude material to give a white powder (yield: Methanol = 10:1).
1H NMR(300MHz,DMSO-d6)δ9.08(brs,1H),7.88(brs,1H),7.67(brs,1H),7.56(dd,J＝8.4,2.5Hz,1H),7.52(dd,J＝8.0,2.5Hz,1H),3.73(s,1H),3.21(t,J＝4.0Hz,1H),2.84–2.70(m,1H),2.66–2.54(m,2H),1.07(s,9H)；13C NMR(125MHz,DMSO-d6)δ165.40,158.71(d,J＝235.0Hz,C-F),149.29,140.61,139.06,138.61,136.02,128.94,123.26,110.34(d,J＝30.3Hz,CH-CF),101.80(d,J＝25.4Hz,CH-CF),64.77,43.02,35.89,27.42,27.35；HRMS(ESI):m/z Calcd.For C19H22FN4OS[M+H]+:373.1498；Found: 373.1490；[α]20D–191(c,0.08,MeOH)；ee>99％；tR＝12.69min,色谱条件为：色谱柱型号：Chiralpak AD-H；色谱柱规格：0.40cm I.D.×25cm L；进样量：5μL；流动相：Hexane/IPA/DEA＝70/30/0.05(v/v/v)；流速：0.40ml/min；检测条件：UVλ＝254nm；柱温：20℃。 1 H NMR (300MHz, DMSO- d 6) δ9.08 (brs, 1H), 7.88 (brs, 1H), 7.67 (brs, 1H), 7.56 (dd, J = 8.4,2.5Hz, 1H), 7.52 ( Dd, J = 8.0, 2.5 Hz, 1H), 3.73 (s, 1H), 3.21 (t, J = 4.0 Hz, 1H), 2.84 - 2.70 (m, 1H), 2.66 - 2.54 (m, 2H), 1.07 (s, 9H); 13 C NMR (125MHz, DMSO-d 6 ) δ 165.40, 158.71 (d, J = 235.0 Hz, CF), 149.29, 140.61, 139.06, 138.61, 136.02, 128.94, 123.26, 110.34 (d, J =30.3 Hz, CH-CF), 101.80 (d, J = 25.4 Hz, CH-CF), 64.77, 43.02, 35.89, 27.42, 27.35; HRMS (ESI): m/z Calcd. For C 19 H 22 FN 4 OS[M+H] + :373.1498;Found: 373.1490;[α]20D–191(c,0.08,MeOH); ee>99%; t R =12.69 min, chromatographic conditions: column type: Chiralpak AD- H; column size: 0.40 cm ID × 25 cm L; injection amount: 5 μL; mobile phase: Hexane / IPA / DEA = 70 / 30 / 0.05 (v / v / v); flow rate: 0.40 ml / min; detection conditions : UV λ = 254 nm; column temperature: 20 ° C.
实施例25. (R)-2-(7-叔丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺，I-25的制备：Example 25. (R)-2-(7-tert-Butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H-benzene And [d] imidazole-4-carboxamide, preparation of I-25:
步骤1. (R)-2-(7-叔丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯：Step 1. (R)-2-(7-tert-Butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H-benzo [d] Methyl imidazole-4-carboxylate:
Figure PCTCN2016087368-appb-000126
Figure PCTCN2016087368-appb-000126
分离条件如S-4-13。The separation conditions are as in S-4-13.
1H NMR(300MHz,Chloroform-d)δ10.40(s,1H),7.63(dd,J＝15.5,1.8Hz,1H),7.60(dd,J＝15.8,1.7Hz,1H),7.39(s,1H),4.02(s,3H),3.84(s,1H),3.43–3.31(m,1H),2.98–2.85(m,1H),2.80–2.58(m,2H),1.13(s,9H)；MS(ESI)388[M+H]+；[α]20D 141(c,0.2,MeOH)；tR＝1.93min。 1 H NMR (300MHz, Chloroform- d) δ10.40 (s, 1H), 7.63 (dd, J = 15.5,1.8Hz, 1H), 7.60 (dd, J = 15.8,1.7Hz, 1H), 7.39 (s , 1H), 4.02 (s, 3H), 3.84 (s, 1H), 3.43 - 3.31 (m, 1H), 2.98 - 2.85 (m, 1H), 2.80 - 2.58 (m, 2H), 1.13 (s, 9H) ); MS (ESI) 388 [ M + H] +; [α] 20D 141 (c, 0.2, MeOH); t R = 1.93min.
步骤2. (R)-2-(7-叔丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺(I-25)：Step 2. (R)-2-(7-tert-Butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1H-benzo [d]Imidazole-4-carboxamide (I-25):
Figure PCTCN2016087368-appb-000127
Figure PCTCN2016087368-appb-000127
按照制备化合物I-1的操作方法，以(R)-2-(7-叔丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯(R-4-13)(200mg，0.52mmol)为原料氨解得到白色粉末125mg，产率为65％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, (R)-2-(7-tert-butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)- 6-Fluoro-1H-benzo[d]imidazole-4-carboxylic acid methyl ester (R-4-13) (200 mg, 0.52 mmol) was obtained as a crude material to give a white powder (yield: Methanol = 10:1).
1H NMR(300MHz,DMSO-d6)δ9.08(brs,1H),7.88(brs,1H),7.67(brs,1H),7.56(dd,J＝8.4,2.5Hz,1H),7.52(dd,J＝8.0,2.5Hz,1H),3.73(s,1H),3.21(t,J＝4.0Hz,1H),2.84–2.70(m,1H),2.66–2.54(m,2H),1.07(s,9H)；13C NMR(125MHz,DMSO-d6)δ165.40,158.71(d,J＝235.0Hz,C-F),149.29,140.61,139.06,138.61,136.02,128.94,123.26,110.34(d,J＝30.3Hz,CH-CF),101.80(d,J＝25.4Hz,CH-CF),64.77,43.02,35.89,27.42,27.35；HRMS(ESI):m/z Calcd.For C19H22FN4OS[M+H]+:373.1498；Found:373.1490；[α]20D 190(c,0.09,MeOH)；ee＝98％；tR＝13.42min,色谱条件为：色谱柱型 号：Chiralpak AD-H；色谱柱规格：0.40cm I.D.×25cm L；进样量：5μL；流动相：Hexane/IPA/DEA＝70/30/0.05(v/v/v)；流速：0.40ml/min；检测条件：UVλ＝254nm；柱温：20℃。 1 H NMR (300MHz, DMSO- d 6) δ9.08 (brs, 1H), 7.88 (brs, 1H), 7.67 (brs, 1H), 7.56 (dd, J = 8.4,2.5Hz, 1H), 7.52 ( Dd, J = 8.0, 2.5 Hz, 1H), 3.73 (s, 1H), 3.21 (t, J = 4.0 Hz, 1H), 2.84 - 2.70 (m, 1H), 2.66 - 2.54 (m, 2H), 1.07 (s, 9H); 13 C NMR (125MHz, DMSO-d 6 ) δ 165.40, 158.71 (d, J = 235.0 Hz, CF), 149.29, 140.61, 139.06, 138.61, 136.02, 128.94, 123.26, 110.34 (d, J =30.3 Hz, CH-CF), 101.80 (d, J = 25.4 Hz, CH-CF), 64.77, 43.02, 35.89, 27.42, 27.35; HRMS (ESI): m/z Calcd. For C 19 H 22 FN 4 OS[M+H] + : 373.1498; Found: 373.1490; [α] 20D 190 (c, 0.09, MeOH); ee = 98%; t R = 13.42 min, chromatographic conditions: column type: Chiralpak AD-H Column specification: 0.40 cm ID × 25 cm L; injection volume: 5 μL; mobile phase: Hexane/IPA/DEA = 70/30/0.05 (v/v/v); flow rate: 0.40 ml/min; detection conditions: UV λ = 254 nm; column temperature: 20 ° C.
实施例26. 6-氟-2-(7-异丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-26的制备：Example 26. 6-Fluoro-2-(7-isobutyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d] Preparation of imidazole-4-carboxamide, I-26:
步骤1. 3-甲基-N-(2-(噻吩-3-基)乙基)丁酰胺：Step 1. 3-Methyl-N-(2-(thien-3-yl)ethyl)butanamide:
Figure PCTCN2016087368-appb-000128
Figure PCTCN2016087368-appb-000128
按照制备16-1的操作方法，以2-(噻吩-3-基)乙基胺盐酸盐(492mg，3.0mmol)，3-甲基丁酰氯(434mg，3.6mmol)为原料制得浅黄色油状物627mg，产率为99％，直接进入下一步反应。According to the procedure of Preparation 16-1, 2-(thiophen-3-yl)ethylamine hydrochloride (492 mg, 3.0 mmol), 3-methylbutyryl chloride (434 mg, 3.6 mmol) The oil was 627 mg in a yield of 99% and was directly passed to the next reaction.
1H NMR(300MHz,Chloroform-d)δ7.28(dd,J＝4.9,3.0Hz,1H),7.01–6.98(m,1H),6.95(dd,J＝1.3Hz,1H),5.44(brs,1H),3.57–3.48(m,2H),2.84(t,J＝6.8Hz,2H),2.19–2.01(m,1H),2.01–1.96(m,2H),0.92(d,J＝6.4Hz,6H)；MS(ESI)212[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 7.28 (dd, J = 4.9, 3.0 Hz, 1H), 7.01 - 6.98 (m, 1H), 6.95 (dd, J = 1.3 Hz, 1H), 5.44 (brs) , 1H), 3.57–3.48 (m, 2H), 2.84 (t, J = 6.8 Hz, 2H), 2.19–2.01 (m, 1H), 2.01–1.96 (m, 2H), 0.92 (d, J=6.4) Hz, 6H); MS (ESI) 212 [M+H] + .
步骤2. 7-异丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 2. 7-Isobutyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000129
Figure PCTCN2016087368-appb-000129
按照制备14-3的操作方法，以N-(2-(噻吩-3-基)乙基)异丁酰胺(16-8)(627mg，3.0mmol)为原料制得白色固体816mg，三步总产率为92％(石油醚：乙酸乙酯＝30：1)。According to the procedure of Preparation 14-3, N-(2-(thiophen-3-yl)ethyl)isobutyramide (16-8) (627 mg, 3.0 mmol) was used as the starting material to obtain 816 mg of white solid, three-step total The yield was 92% (petroleum ether: ethyl acetate = 30:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ7.13(d,J＝5.1Hz,1H),6.76(d,J＝5.1Hz,1H),5.48–5.33(m,1H),4.23–4.04(m,1H),3.13–2.95(m,2H),1.89–1.66(m,3H),1.47(s,9H),1.47(d,J＝1.6Hz,10H),1.04(d,J＝6.2Hz,3H),0.96(d,J＝6.3Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ7.13(d,J＝5.1Hz,1H),6.76(d,J＝5.1Hz,1H),5.29–5.15(m,1H),4.43–4.27(m,1H),2.84–2.64(m,2H),1.89–1.66(m,3H),1.47(s,9H),1.04(d,J＝6.2Hz,3H),1.04(d,J＝6.2Hz,3H),0.96(d,J＝6.3Hz,3H)；MS(ESI)318[M+Na]+。Conformational isomer 1: 1 H NMR (300 MHz, Chloroform-d) δ 7.13 (d, J = 5.1 Hz, 1H), 6.76 (d, J = 5.1 Hz, 1H), 5.48 - 5.33 (m, 1H) , 4.23–4.04 (m, 1H), 3.13–2.95 (m, 2H), 1.89–1.66 (m, 3H), 1.47 (s, 9H), 1.47 (d, J=1.6 Hz, 10H), 1.04 (d) , J = 6.2 Hz, 3H), 0.96 (d, J = 6.3 Hz, 3H); conformational isomer 2: 1 H NMR (300 MHz, Chloroform-d) δ 7.13 (d, J = 5.1 Hz, 1H) , 6.76 (d, J = 5.1 Hz, 1H), 5.29 - 5.15 (m, 1H), 4.43 - 4.27 (m, 1H), 2.84 - 2.64 (m, 2H), 1.89 - 1.66 (m, 3H), 1.47 (s, 9H), 1.04 (d, J = 6.2 Hz, 3H), 1.04 (d, J = 6.2 Hz, 3H), 0.96 (d, J = 6.3 Hz, 3H); MS (ESI) 318 [M+ Na] + .
步骤3. 2-溴-7-异丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯： Step 3. 2-Bromo-7-isobutyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000130
Figure PCTCN2016087368-appb-000130
按照制备12-3的操作方式，以7-异丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(14-10)(816mg，2.8mmol)为原料经溴代制得664mg无色油状物，产率为64％(石油醚：乙酸乙酯＝30：1)。According to the procedure of Preparation 12-3, 7-isobutyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (14-10) (816 mg) </RTI> 2.8 mmol) 264 mg of EtOAc (yield: EtOAc (EtOAc)
构象异构体1：1H NMR(300MHz,Chloroform-d)δ6.71(s,1H),5.38–5.20(m,1H),4.20–3.92(m,1H),3.19–2.89(m,2H),1.73(hept,J＝6.3Hz,1H),1.47(s,9H),1.02(d,J＝6.3Hz,3H),0.95(d,J＝6.4Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ6.71(s,1H),5.16–4.97(m,1H),4.42–4.23(m,1H),2.80–2.42(m,2H),1.73(hept,J＝6.3Hz,1H),1.47(s,9H),1.02(d,J＝6.3Hz,3H),0.95(d,J＝6.4Hz,3H)；MS(ESI)396[M+Na]+。Conformation isomer 1 : 1 H NMR (300 MHz, Chloroform-d) δ 6.71 (s, 1H), 5.38 - 5.20 (m, 1H), 4.20 - 3.92 (m, 1H), 3.19 - 2.89 (m, 2H) ), 1.73 (hept, J = 6.3 Hz, 1H), 1.47 (s, 9H), 1.02 (d, J = 6.3 Hz, 3H), 0.95 (d, J = 6.4 Hz, 3H); conformation 2 : 1 H NMR (300 MHz, Chloroform-d) δ 6.71 (s, 1H), 5.16 - 4.97 (m, 1H), 4.42 - 4.23 (m, 1H), 2.80 - 2.42 (m, 2H), 1.73 (hept , J = 6.3 Hz, 1H), 1.47 (s, 9H), 1.02 (d, J = 6.3 Hz, 3H), 0.95 (d, J = 6.4 Hz, 3H); MS (ESI) 396 [M+Na] + .
步骤4. 2-甲酰基-7-异丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 4. 2-Formyl-7-isobutyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000131
Figure PCTCN2016087368-appb-000131
按照制备化合物2-3的操作方法，以2-溴-7-异丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(12-10)(664mg，1.8mmol)为原料制备得到黄色油状物400mg，产率为69％(石油醚：乙酸乙酯＝8：1)。According to the procedure for the preparation of compound 2-3, 2-bromo-7-isobutyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (12) -10) (664 mg, 1.8 mmol) was obtained as a crude material (yield:ield:
构象异构体1：1H NMR(300MHz,Chloroform-d)δ9.83(s,1H),7.44(s,1H),5.45(brs,1H),4.24–4.12(m,1H),3.19–2.70(m,2H),2.68–2.52(m,1H),1.86–1.70(m,2H),1.48(s,9H),1.05(d,J＝6.4Hz,3H),0.97(d,J＝6.4Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ9.83(s,1H),7.44(s,1H),5.25(brs,1H),4.44–4.33(m,1H),3.19–2.70(m,2H),2.68–2.52(m,1H),1.86–1.70(m,2H),1.48(s,9H),1.05(d,J＝6.4Hz,3H),0.97(d,J＝6.4Hz,3H)；MS(ESI)346[M+Na]+。Conformation isomer 1 : 1 H NMR (300 MHz, Chloroform-d) δ 9.83 (s, 1H), 7.44 (s, 1H), 5.45 (brs, 1H), 4.24 - 4.12 (m, 1H), 3.19 - 2.70 (m, 2H), 2.68 - 2.52 (m, 1H), 1.86 - 1.70 (m, 2H), 1.48 (s, 9H), 1.05 (d, J = 6.4 Hz, 3H), 0.97 (d, J = 6.4 Hz, 3H); conformational isomer 2: 1 H NMR (300 MHz, Chloroform-d) δ 9.83 (s, 1H), 7.44 (s, 1H), 5.25 (brs, 1H), 4.44 - 4.33 (m) , 1H), 3.19–2.70 (m, 2H), 2.68–2.52 (m, 1H), 1.86–1.70 (m, 2H), 1.48 (s, 9H), 1.05 (d, J = 6.4 Hz, 3H), 0.97 (d, J = 6.4 Hz, 3H); MS (ESI) 346 [M+Na] + .
步骤5. 2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-7-异丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 5. 2-(6-Fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-7-isobutyl-4,7-dihydrothieno[2, 3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000132
Figure PCTCN2016087368-appb-000132
按照制备化合物3-7的操作方法，以2-甲酰基-7-异丁基-4,7-二氢噻吩并[2,3-c]吡啶 -6(5H)-羧酸叔丁酯(2-10)(400mg，1.24mmol)和2,3-二氨基-5-氟苯甲酸甲酯(228mg，1.24mmol)为原料制备得到淡黄色固体553mg，产率为91％(石油醚：乙酸乙酯＝3：1)。According to the procedure for the preparation of compound 3-7, 2-formyl-7-isobutyl-4,7-dihydrothieno[2,3-c]pyridine -6(5H)-tert-butyl carboxylic acid tert-butyl ester (2-10) (400 mg, 1.24 mmol) and 2,3-diamino-5-fluorobenzoic acid methyl ester (228 mg, 1.24 mmol) were obtained as a pale yellow solid. 553 mg, yield 91% (petroleum ether: ethyl acetate = 3:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ7.64(dd,J＝9.2,2.4Hz,1H),7.62(dd,J＝9.2,2.4Hz,1H),7.39(s,1H),5.45(brs,1H),4.30–4.12(m,1H),4.02(s,3H),2.91–2.70(m,2H),2.69–2.54(m,1H),1.95–1.68(m,2H),1.67–1.50(m,1H),1.50(s,9H),1.06(d,J＝6.4Hz,3H),1.01(d,J＝6.5Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ7.64(dd,J＝9.2,2.4Hz,1H),7.62(dd,J＝9.2,2.4Hz,1H),7.39(s,1H),5.25(brs,1H),4.50–4.32(m,1H),4.02(s,3H),3.20–2.96(m,2H),2.69–2.54(m,1H),1.95–1.68(m,2H),1.67–1.50(m,1H),1.50(s,9H),1.06(d,J＝6.4Hz,3H),1.01(d,J＝6.5Hz,3H)；MS(ESI)488[M+H]+。Conformational isomer 1: 1 H NMR (300 MHz, Chloroform-d) δ 7.64 (dd, J = 9.2, 2.4 Hz, 1H), 7.62 (dd, J = 9.2, 2.4 Hz, 1H), 7.39 (s, 1H), 5.45 (brs, 1H), 4.30–4.12 (m, 1H), 4.02 (s, 3H), 2.91–2.70 (m, 2H), 2.69–2.54 (m, 1H), 1.95–1.68 (m, 2H), 1.67 - 1.50 (m, 1H), 1.50 (s, 9H), 1.06 (d, J = 6.4 Hz, 3H), 1.01 (d, J = 6.5 Hz, 3H); conformation 2: 1 H NMR (300 MHz, Chloroform-d) δ 7.64 (dd, J = 9.2, 2.4 Hz, 1H), 7.62 (dd, J = 9.2, 2.4 Hz, 1H), 7.39 (s, 1H), 5.25 (brs, 1H), 4.50–4.32 (m, 1H), 4.02 (s, 3H), 3.20–2.96 (m, 2H), 2.69–2.54 (m, 1H), 1.95–1.68 (m, 2H), 1.67–1.50 ( m, 1H), 1.50 (s, 9H), 1.06 (d, J = 6.4 Hz, 3H), 1.01 (d, J = 6.5 Hz, 3H); MS (ESI) 488 [M+H] + .
步骤6. 6-氟-2-(7-异丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 6. 6-Fluoro-2-(7-isobutyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole -4-methyl formate:
Figure PCTCN2016087368-appb-000133
Figure PCTCN2016087368-appb-000133
按照制备化合物4-7的操作方法，以2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-7-异丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(3-14)(553mg，1.13mmol)为原料去保护基得到白色粉末374mg，产率为85％(二氯甲烷：甲醇＝50：1)。According to the procedure for the preparation of compound 4-7, 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-7-isobutyl-4,7 -Dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (3-14) (553 mg, 1.13 mmol) as a starting material deprotecting group afforded white powder 374mg, yield 85% (Dichloromethane: methanol = 50:1).
1H NMR(300MHz,Chloroform-d)δ10.42(brs,1H),7.62(dd,J＝13.3,2.4Hz,1H),7.59(dd,J＝14.1,2.4Hz,1H),7.38(s,1H),4.17–4.03(m,1H),4.02(s,3H),3.34(ddd,J＝13.0,5.3,3.7Hz,1H),3.01(ddd,J＝13.0,8.7,5.6Hz,1H),2.75–2.65(m,2H),2.11–1.91(m,1H),1.67–1.62(m,2H),1.02(d,J＝4.2Hz,3H),1.00(d,J＝4.3Hz,3H)；MS(ESI)388[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ10.42 (brs, 1H), 7.62 (dd, J = 13.3,2.4Hz, 1H), 7.59 (dd, J = 14.1,2.4Hz, 1H), 7.38 (s , 1H), 4.17 - 4.03 (m, 1H), 4.02 (s, 3H), 3.34 (ddd, J = 13.0, 5.3, 3.7 Hz, 1H), 3.01 (ddd, J = 13.0, 8.7, 5.6 Hz, 1H ), 2.75–2.65 (m, 2H), 2.11–1.91 (m, 1H), 1.67–1.62 (m, 2H), 1.02 (d, J=4.2 Hz, 3H), 1.00 (d, J=4.3 Hz, 3H); MS (ESI) 388 [M+H] + .
步骤7. 6-氟-2-(7-异丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-26)：Step 7. 6-Fluoro-2-(7-isobutyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole -4-carboxamide (I-26):
Figure PCTCN2016087368-appb-000134
Figure PCTCN2016087368-appb-000134
按照制备化合物I-1的操作方法，以6-氟-2-(7-异丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(4-14)(374mg，0.97mmol)为原料氨解得到淡黄色粉末241mg，产率为67％(二氯甲烷：甲醇＝10：1)。 According to the procedure for the preparation of compound I-1, 6-fluoro-2-(7-isobutyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)- 1H-Benzo[d]imidazole-4-carboxylic acid methyl ester (4-14) (374 mg, 0.97 mmol) was obtained as a crude material to give a pale yellow powder 241 mg (yield: 67%) ).
1H NMR(500MHz,DMSO-d6)δ9.01(brs,1H),7.92(brs,1H),7.62(s,1H),7.53(dd,J＝10.7,2.6Hz,1H),7.50(dd,J＝8.5,2.6Hz,1H),3.94(dd,J＝10.1,4.3Hz,1H),3.11(dt,J＝13.1,5.4Hz,1H),2.81(ddd,J＝13.4,8.7,5.4Hz,1H),2.63-2.52(m,2H),1.99–1.89(m,1H),1.58(ddd,J＝14.2,9.7,4.7Hz,1H),1.48(ddd,J＝13.6,9.2,4.3Hz,1H),0.93(d,J＝6.6Hz,3H),0.92(d,J＝6.6Hz,3H)；13C NMR(125MHz,DMSO-d6)δ165.46,158.6(d,J＝237.5Hz,C-F),149.45,145.03,136.18,129.01,128.49,122.76,110.11(d,J＝25.8Hz,CH-CF),102.11(d,J＝23.4Hz,CH-CF),52.54,46.85,41.76,26.57,24.56,23.87,21.91；HRMS(ESI):m/z Calcd.For C19H22FN4OS[M+H]+:373.1498；Found:373.1490。 1 H NMR (500MHz, DMSO- d 6) δ9.01 (brs, 1H), 7.92 (brs, 1H), 7.62 (s, 1H), 7.53 (dd, J = 10.7,2.6Hz, 1H), 7.50 ( Dd, J = 8.5, 2.6 Hz, 1H), 3.94 (dd, J = 10.1, 4.3 Hz, 1H), 3.11 (dt, J = 13.1, 5.4 Hz, 1H), 2.81 (ddd, J = 13.4, 8.7, 5.4 Hz, 1H), 2.63-2.52 (m, 2H), 1.99–1.89 (m, 1H), 1.58 (ddd, J=14.2, 9.7, 4.7 Hz, 1H), 1.48 (ddd, J=13.6, 9.2, 4.3 Hz, 1H), 0.93 (d, J = 6.6 Hz, 3H), 0.92 (d, J = 6.6 Hz, 3H); 13 C NMR (125 MHz, DMSO-d 6 ) δ 165.46, 158.6 (d, J = 237.5) Hz, CF), 149.45, 145.03, 136.18, 129.01, 128.49, 122.76, 110.11 (d, J = 25.8 Hz, CH-CF), 102.11 (d, J = 23.4 Hz, CH-CF), 52.54, 46.85, 41.76 , 26.57, 24.56, 23.87, 21.91; HRMS (ESI): m/z Calcd. For C 19 H 22 FN 4 OS [M+H] + : 373.1498; Found: 373.1490.
实施例27. 6-氟-2-(5-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-27的制备：Example 27. 6-Fluoro-2-(5-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole Preparation of 4-carboxamide, I-27:
步骤1. 3-(2-硝基丙基-1-烯-1-基)噻吩：Step 1. 3-(2-Nitropropyl-1-en-1-yl)thiophene:
Figure PCTCN2016087368-appb-000135
Figure PCTCN2016087368-appb-000135
在冰浴下，向噻吩-3-甲醛(18)(2.24g，20.0mmol)和硝基乙烷(60mL)的混合物中加入醋酸铵(2.00g，26.0mmol)，回流两小时，将反应液减压浓缩，重新溶于50mL乙酸乙酯中，依次水洗，饱和食盐水洗，无水硫酸钠干燥，过滤减压浓缩制得淡棕色固体2.87g，产率为85％。Ammonium acetate (2.00 g, 26.0 mmol) was added to a mixture of thiophene-3-carbaldehyde (18) (2.24 g, 20.0 mmol) and nitroethane (60 mL), and the mixture was refluxed for two hours. The organic layer was concentrated under reduced pressure. EtOAc was evaporated, evaporated, evaporated.
1H NMR(300MHz,Chloroform-d)δ8.09(s,1H),7.60(brs,1H),7.44(dd,J＝5.0,3.0Hz,1H),7.27(d,J＝4.6Hz,1H),2.50(s,3H)；MS(ESI)170[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 8.09 (s, 1H), 7.60 (brs, 1H), 7.44 (dd, J = 5.0, 3.0 Hz, 1H), 7.27 (d, J = 4.6 Hz, 1H) ), 2.50 (s, 3H); MS (ESI) 170 [M+H] + .
步骤2. (1-(噻吩-3-基)丙基-2-基)氨基甲酸叔丁酯：Step 2. (1-(Thien-3-yl)propyl-2-yl)carbamic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000136
Figure PCTCN2016087368-appb-000136
0℃下，往溶有3-(2-硝基丙基-1-烯-1-基)噻吩(19)(1.70g，10.0mmol)中的无水四氢呋喃(10mL)溶液中加入氢化铝锂四氢呋喃溶液(2.5M)(12.5mL)，缓慢升至60℃反应1小时，冰浴冷却，缓慢加入五水硫酸钠淬灭，二氯甲烷萃取(50mL×3)，水洗，饱和食盐水洗，保留有机层。0℃下往有机层中缓慢加入二碳酸二叔丁酯(1.78g，10.0mmol)，移至室温搅拌1小时，减压蒸去大部分溶剂，经柱层析得淡黄色油状物1.33g，产率为55％。Add lithium aluminum hydride to a solution of 3-(2-nitropropyl-1-en-1-yl)thiophene (19) (1.70 g, 10.0 mmol) in anhydrous tetrahydrofuran (10 mL) at 0 ° C Tetrahydrofuran solution (2.5M) (12.5mL), slowly rise to 60 ° C for 1 hour, cooled in an ice bath, slowly added with sodium sulfate pentahydrate, quenched, extracted with dichloromethane (50mL × 3), washed with water, washed with saturated brine, retained Organic layer. Di-tert-butyl dicarbonate (1.78 g, 10.0 mmol) was slowly added to the organic layer at 0 ° C, and the mixture was stirred at room temperature for 1 hour, and most of the solvent was evaporated under reduced pressure. The yield was 55%.
1H NMR(300MHz,Chloroform-d)δ7.26(dd,J＝4.7,2.9Hz,1H),7.00–6.96(m,1H),6.94(dd,J＝4.9,1.1Hz,1H),4.36(brs,1H),3.91(brs,1H),2.78(qd,J＝14.1,6.1Hz,1H),1.43(s,9H),1.09(d,J＝6.6Hz,3H)；MS(ESI)264[M+Na]+。 1 H NMR (300MHz, Chloroform- d) δ7.26 (dd, J = 4.7,2.9Hz, 1H), 7.00-6.96 (m, 1H), 6.94 (dd, J = 4.9,1.1Hz, 1H), 4.36 (brs, 1H), 3.91 (brs, 1H), 2.78 (qd, J = 14.1, 6.1 Hz, 1H), 1.43 (s, 9H), 1.09 (d, J = 6.6 Hz, 3H); MS (ESI) 264[M+Na] + .
步骤3. 5-甲基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯： Step 3. 5-Methyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000137
Figure PCTCN2016087368-appb-000137
向溶有叔丁基(1-(噻吩-3-基)丙基-2-基)氨基甲酸酯(21)(500mg，2.07mmol)的甲苯溶液中加入多聚甲醛(150mg，5.00mmol)，一水合对甲苯磺酸(19mg，0.10mmol)，回流过夜，减压蒸去大部分溶剂，粗产品经柱层析得到淡黄色油状物257mg，产率为49％。Add paraformaldehyde (150 mg, 5.00 mmol) to a solution of tert-butyl (1-(thien-3-yl)propyl-2-yl)carbamate (21) (500 mg, 2.07 mmol) in toluene The p-toluenesulfonic acid monohydrate (19 mg, 0.10 mmol) was refluxed overnight.
1H NMR(300MHz,Chloroform-d)δ7.15(d,J＝5.0Hz,1H),6.77(d,J＝5.0Hz,1H),4.97(d,J＝16.9Hz,1H),4.75(brs,1H),4.19(d,J＝16.8Hz,1H),2.93(dd,J＝16.0,6.5Hz,1H),2.50(d,J＝15.6Hz,1H),1.48(s,9H),1.12(d,J＝6.9Hz,3H)；MS(ESI)276[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ7.15 (d, J = 5.0Hz, 1H), 6.77 (d, J = 5.0Hz, 1H), 4.97 (d, J = 16.9Hz, 1H), 4.75 ( Brs, 1H), 4.19 (d, J = 16.8 Hz, 1H), 2.93 (dd, J = 16.0, 6.5 Hz, 1H), 2.50 (d, J = 15.6 Hz, 1H), 1.48 (s, 9H), 1.12 (d, J = 6.9 Hz, 3H); MS (ESI) 276 [M+H] + .
步骤4. 2-溴-5-甲基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 4. 2-Bromo-5-methyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000138
Figure PCTCN2016087368-appb-000138
按照制备12-3的操作方式，以5-甲基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(14-11)(257mg，1.02mmol)为原料经溴代制得249mg淡黄色油状物，产率为75％(石油醚：乙酸乙酯＝30：1)。According to the procedure of Preparation 12-3, 5-methyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (14-11) (257 mg, 1.02 mmol) was obtained by bromination of 249 mg of pale yellow oil (yield: petroleum ether: ethyl acetate = 30:1).
1H NMR(300MHz,Chloroform-d)δ6.73(s,1H),4.84(d,J＝17.2Hz,1H),4.70(brs,1H),4.07(d,J＝17.0Hz,1H),2.87(ddt,J＝16.1,5.5,2.4Hz,1H),2.40(dt,J＝15.9,2.4Hz,1H),1.47(s,9H),1.12(d,J＝6.9Hz,3H)；MS(ESI)354[M+Na]+。 1 H NMR (300MHz, Chloroform- d) δ6.73 (s, 1H), 4.84 (d, J = 17.2Hz, 1H), 4.70 (brs, 1H), 4.07 (d, J = 17.0Hz, 1H), 2.87 (ddt, J = 16.1, 5.5, 2.4 Hz, 1H), 2.40 (dt, J = 15.9, 2.4 Hz, 1H), 1.47 (s, 9H), 1.12 (d, J = 6.9 Hz, 3H); MS (ESI) 354 [M+Na] + .
步骤5. 2-甲酰基-5-甲基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 5. 2-Formyl-5-methyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000139
Figure PCTCN2016087368-appb-000139
按照制备化合物2-3的操作方法，以2-溴-5-甲基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(12-11)(249mg，0.98mmol)为原料制备得到淡黄色油状物134mg，产率为48％(石油醚：乙酸乙酯＝8：1)。According to the procedure for the preparation of compound 2-3, 2-bromo-5-methyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (12- 11) (249 mg, 0.98 mmol) was obtained as a crude material (yield: 134 mg,yield: 48%).
1H NMR(300MHz,Chloroform-d)δ9.85(s,1H),7.48(s,1H),5.04(d,J＝18.3Hz,1H),4.78(brs,1H),4.24(d,J＝18.3Hz,1H),2.97(dd,J＝15.8,5.9Hz,1H),2.54(d,J＝15.9Hz,1H),1.49(s,9H),1.12(d,J＝6.9Hz,3H)；MS(ESI)282[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 9.85 (s, 1H), 7.48 (s, 1H), 5.04 (d, J = 18.3 Hz, 1H), 4.78 (brs, 1H), 4.24 (d, J) =18.3 Hz, 1H), 2.97 (dd, J = 15.8, 5.9 Hz, 1H), 2.54 (d, J = 15.9 Hz, 1H), 1.49 (s, 9H), 1.12 (d, J = 6.9 Hz, 3H) MS (ESI) 282 [M+H] + .
步骤6. 2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-5-甲基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯： Step 6. 2-(6-Fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-5-methyl-4,7-dihydrothieno[2,3 -c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000140
Figure PCTCN2016087368-appb-000140
按照制备化合物3-7的操作方法，以2-甲酰基-5-甲基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(2-11)(134mg，0.47mmol)和2,3-二氨基-5-氟苯甲酸甲酯(86mg，0.47mmol)为原料制备得到淡黄色固体187mg，产率为89％(石油醚：乙酸乙酯＝3：1)。According to the procedure for the preparation of compound 3-7, 2-formyl-5-methyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (2 -11) (134 mg, 0.47 mmol) and 2,3-diamino-5-fluorobenzoic acid methyl ester (86 mg, 0.47 mmol) were used as starting materials to give 187 mg of pale yellow solid (yield: 89%). Ester = 3:1).
1H NMR(300MHz,Chloroform-d)δ10.40(s,1H),7.63(dd,J＝13.4,2.1Hz,1H),7.60(dd,J＝14.4,2.3Hz,1H),7.38(s,1H),5.11-4.97(d,J＝16.0,1H),4.80(brs,1H),4.31-4.19(m,1H),4.02(s,3H),2.97(d,J＝15.8,5.7Hz,1H),2.55(d,J＝15.6Hz,1H),1.50(s,9H),1.16(d,J＝7.0Hz,3H)；MS(ESI)446[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ10.40 (s, 1H), 7.63 (dd, J = 13.4,2.1Hz, 1H), 7.60 (dd, J = 14.4,2.3Hz, 1H), 7.38 (s , 1H), 5.11-4.97 (d, J = 16.0, 1H), 4.80 (brs, 1H), 4.31-4.19 (m, 1H), 4.02 (s, 3H), 2.97 (d, J = 15.8, 5.7 Hz , 1H), 2.55 (d, J = 15.6 Hz, 1H), 1.50 (s, 9H), 1.16 (d, J = 7.0 Hz, 3H); MS (ESI) 446 [M+H] + .
步骤7. 6-氟-2-(5-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 7. 6-Fluoro-2-(5-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole- 4-methyl formate:
Figure PCTCN2016087368-appb-000141
Figure PCTCN2016087368-appb-000141
按照制备化合物4-7的操作方法，以2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-5-甲基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(3-15)(187mg，0.42mmol)为原料去保护基得到白色粉末119mg，产率为82％(二氯甲烷：甲醇＝50：1)。According to the procedure for the preparation of compound 4-7, 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-5-methyl-4,7- Dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (3-15) (187 mg, 0.42 mmol) was used as a starting material to give a white powder, y. Dichloromethane: methanol = 50:1).
1H NMR(300MHz,Chloroform-d)δ10.38(brs,1H),7.63(dd,J＝12.5,2.3Hz,1H),7.59(dd,J＝14.0,2.3Hz,1H),7.37(s,1H),4.16(s,2H),4.02(s,3H),3.13-3.00(m,1H),2.79(dd,J＝16.2,4.0Hz,1H),2.38(dd,J＝16.1,10.1Hz,1H),1.30(d,J＝6.4Hz,3H)；MS(ESI)346[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ10.38 (brs, 1H), 7.63 (dd, J = 12.5,2.3Hz, 1H), 7.59 (dd, J = 14.0,2.3Hz, 1H), 7.37 (s , 1H), 4.16 (s, 2H), 4.02 (s, 3H), 3.13-3.00 (m, 1H), 2.79 (dd, J = 16.2, 4.0 Hz, 1H), 2.38 (dd, J = 16.1, 10.1 Hz, 1H), 1.30 (d, J = 6.4 Hz, 3H); MS (ESI) 346 [M+H] + .
步骤8. 6-氟-2-(5-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-27)：Step 8. 6-Fluoro-2-(5-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole- 4-carboxamide (I-27):
Figure PCTCN2016087368-appb-000142
Figure PCTCN2016087368-appb-000142
按照制备化合物I-1的操作方法，以6-氟-2-(5-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶 -2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(4-15)(119mg，0.34mmol)为原料氨解得到淡黄色粉末82mg，产率为73％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 6-fluoro-2-(5-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridine The methyl group of 2-yl)-1H-benzo[d]imidazole-4-carboxylate (4-15) (119 mg, 0.34 mmol) was obtained as a crude material to give a pale yellow powder (yield: :Methanol = 10:1).
1H NMR(300MHz,DMSO-d6)δ8.96(brs,1H),7.89(brs,1H),7.67(s,1H),7.57–7.45(m,2H),4.06(d,J＝28.9,1H),4.01(d,J＝29.1,1H),2.99(brs,1H),2.74(dd,J＝16.3,3.9Hz,1H),2.41–2.25(m,1H),1.19(d,J＝6.3Hz,3H)；HRMS(ESI):m/z Calcd.For C16H16FN4OS[M+H]+:331.1029；Found:311.1023。 1 H NMR (300MHz, DMSO- d 6) δ8.96 (brs, 1H), 7.89 (brs, 1H), 7.67 (s, 1H), 7.57-7.45 (m, 2H), 4.06 (d, J = 28.9 , 1H), 4.01 (d, J = 29.1, 1H), 2.99 (brs, 1H), 2.74 (dd, J = 16.3, 3.9 Hz, 1H), 2.41 - 2.25 (m, 1H), 1.19 (d, J) = 6.3 Hz, 3H); HRMS (ESI): m/z Calcd. For C 16 H 16 FN 4 OS [M+H] + : 331.1029; Found: 311.1023.
实施例28. 6-氟-2-(3-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-28的制备：Example 28. 6-Fluoro-2-(3-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole Preparation of 4-carboxamide, I-28:
步骤1. 5-叔丁基3-乙基2-氨基-6,7-二氢噻吩并[3,2-c]吡啶-3,5(4H)-二羧酸酯：Step 1. 5-tert-Butyl 3-ethyl 2-amino-6,7-dihydrothieno[3,2-c]pyridine-3,5(4H)-dicarboxylate:
Figure PCTCN2016087368-appb-000143
Figure PCTCN2016087368-appb-000143
向溶有N-叔丁氧羰基-4-哌啶酮(22)(9.95g，50.0mmol)和氰基乙酸乙酯(5.65g，50.0mmol)的无水乙醇(250mL)溶液中加入1.60g硫粉和4.35g吗啡啉，回流1小时，减压浓缩，粗产品在乙醇中经重结晶得到浅棕色固体13.7g，产率为84％。To a solution of N-tert-butoxycarbonyl-4-piperidone (22) (9.95 g, 50.0 mmol) and ethyl cyanoacetate (5.65 g, 50.0 mmol) in dry ethanol (250 mL) The sulfur powder and 4.35 g of morpholine were refluxed for 1 hour, and concentrated under reduced pressure. The crude material was crystallised from ethanol to give a pale brown solid (13.7 g).
1H NMR(300MHz,Chloroform-d)δ6.01(brs,2H),4.35(brs,2H),4.26(q,J＝7.1Hz,2H),3.61(t,J＝5.9Hz,2H),2.80(t,J＝5.9Hz,2H),1.47(s,9H),1.33(t,J＝7.1Hz,3H)；MS(ESI)327[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ6.01 (brs, 2H), 4.35 (brs, 2H), 4.26 (q, J = 7.1Hz, 2H), 3.61 (t, J = 5.9Hz, 2H), 2.80 (t, J = 5.9 Hz, 2H), 1.47 (s, 9H), 1.33 (t, J = 7.1 Hz, 3H); MS (ESI) 327 [M+H] + .
步骤2. 5-叔丁基3-乙基6,7-二氢噻吩并[3,2-c]吡啶-3,5(4H)-二羧酸酯：Step 2. 5-tert-Butyl 3-ethyl 6,7-dihydrothieno[3,2-c]pyridine-3,5(4H)-dicarboxylate:
Figure PCTCN2016087368-appb-000144
Figure PCTCN2016087368-appb-000144
0℃下，往溶有5-叔丁基3-乙基2-氨基-6,7-二氢噻吩并[3,2-c]吡啶-3,5(4H)-二羧酸酯(23)(6.10g，18.7mmol)的四氢呋喃(30mL)溶液中缓慢滴加亚硝酸异戊酯(5.54mL，41.2mmol)，滴加完毕后移至室温搅拌30分钟，继续升温至回流，反应两小时，降至室温，减压浓缩，粗产品经柱层析(石油醚：乙酸乙酯＝10：1)得到白色固体1.88g，产率为32％。At 0 ° C, there was dissolved 5-tert-butyl 3-ethyl 2-amino-6,7-dihydrothieno[3,2-c]pyridine-3,5(4H)-dicarboxylate (23 (6.10g, 18.7mmol) in tetrahydrofuran (30mL) solution was slowly added dropwise isoamyl nitrite (5.54mL, 41.2mmol), after the addition was completed, the mixture was stirred at room temperature for 30 minutes, and the temperature was further increased to reflux for two hours. The mixture was cooled to room temperature and concentrated under reduced pressure. EtOAc (EtOAc:EtOAc:EtOAc
1H NMR(300MHz,Chloroform-d)δ7.97(s,1H),4.60(s,2H),4.29(q,J＝7.1Hz,2H),3.66(t,J＝5.8Hz,2H),2.98(t,J＝5.9Hz,2H),1.48(s,9H),1.35(m,J＝7.1Hz,3H)；MS(ESI)334[M+Na]+。 1 H NMR (300MHz, Chloroform- d) δ7.97 (s, 1H), 4.60 (s, 2H), 4.29 (q, J = 7.1Hz, 2H), 3.66 (t, J = 5.8Hz, 2H), 2.98 (t, J = 5.9 Hz, 2H), 1.48 (s, 9H), 1.35 (m, J = 7.1 Hz, 3H); MS (ESI) 334 [M+Na] + .
步骤3. 3-(羟甲基)-6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯：Step 3. 3-(Hydroxymethyl)-6,7-dihydrothieno[3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000145
Figure PCTCN2016087368-appb-000145
向硼氢化锂(530mg，24.0mmol)的无水四氢呋喃(5mL)混悬液中缓慢滴加5-叔丁基3-乙基6,7-二氢噻吩并[3,2-c]吡啶-3,5(4H)-二羧酸酯(24)(1.88g，6.0mmol)的甲苯(10mL)溶液，加热至100℃回流5小时。减压浓缩，粗产品经柱层析(石油醚：乙酸乙酯＝4：1)得到无色油状物1.02g，产率为63％。To a suspension of lithium borohydride (530 mg, 24.0 mmol) in anhydrous tetrahydrofuran (5 mL) was slowly added dropwise 5-tert-butyl 3-ethyl 6,7-dihydrothieno[3,2-c]pyridine- A solution of 3,5(4H)-dicarboxylate (24) (1.88 g, 6.0 mmol) in toluene (10 mL). The organic layer was concentrated under reduced pressure. EtOAc (EtOAc:EtOAc:
1H NMR(300MHz,Chloroform-d)δ7.08(s,1H),4.61(s,2H),4.58(d,J＝4.6Hz,2H),3.69(t,J＝5.8Hz,2H),2.68(t,J＝5.8Hz,2H),1.48(s,9H)；MS(ESI)292[M+Na]+。 1 H NMR (300 MHz, Chloroform-d) δ 7.08 (s, 1H), 4.61 (s, 2H), 4.58 (d, J = 4.6 Hz, 2H), 3.69 (t, J = 5.8 Hz, 2H), 2.68 (t, J = 5.8 Hz, 2H), 1.48 (s, 9H); MS (ESI) 292 [M+Na] + .
步骤4. 3-(氯甲基)-6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯：Step 4. 3-(Chloromethyl)-6,7-dihydrothieno[3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000146
Figure PCTCN2016087368-appb-000146
0℃下，往溶有3-(羟甲基)-6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯(25)(1.02g，3.8mmol)和三乙胺(1.06mL，7.6mmol)的二氯甲烷(10mL)溶液中缓慢滴加甲磺酰氯(866mg，7.6mmol)，室温搅拌5小时。减压浓缩，粗产品经柱层析(石油醚：乙酸乙酯＝8：1)得到白色固体817mg，产率为75％。At 0 ° C, 3-(hydroxymethyl)-6,7-dihydrothieno[3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl ester (25) (1.02 g, 3.8) was dissolved. Methanesulfonyl chloride (866 mg, 7.6 mmol) was slowly added dropwise to a solution of EtOAc (EtOAc) (EtOAc) The organic layer was concentrated under reduced pressure. EtOAc (EtOAc:EtOAc:EtOAc
1H NMR(300MHz,Chloroform-d)δ7.15(s,1H),4.60(s,2H),4.50(s,2H),3.71(t,J＝5.8Hz,2H),2.71(t,J＝5.7Hz,2H),1.48(s,9H)；MS(ESI)310[M+Na]+。 1 H NMR (300MHz, Chloroform- d) δ7.15 (s, 1H), 4.60 (s, 2H), 4.50 (s, 2H), 3.71 (t, J = 5.8Hz, 2H), 2.71 (t, J = 5.7 Hz, 2H), 1.48 (s, 9H); MS (ESI) 310 [M+Na] + .
步骤5. 3-甲基-6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯：Step 5. 3-Methyl-6,7-dihydrothieno[3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000147
Figure PCTCN2016087368-appb-000147
往3-(氯甲基)-6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯(26)(817mg，2.8mmol)的甲醇(10mL)溶液中加入雷尼镍(80mg)，氮气保护，置换氢气三次，在氢气氛围中加热至60℃，回流4小时。冷却至室温，过滤，减压浓缩，粗产品经柱层析(石油醚：乙酸乙酯＝20：1)得到无色油状物636mg，产率为90％。To 3-(chloromethyl)-6,7-dihydrothieno[3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl ester (26) (817 mg, 2.8 mmol) in methanol (10 mL) Raney nickel (80 mg) was added to the solution, protected with nitrogen, and the hydrogen was replaced three times, heated to 60 ° C in a hydrogen atmosphere, and refluxed for 4 hours. The mixture was cooled to room temperature, filtered, and evaporated.
1H NMR(300MHz,Chloroform-d)δ6.76(s,1H),4.60(s,2H),3.69(t,J＝5.7Hz,2H),2.56(t,J＝5.7Hz,2H),2.11(s,3H),1.48(s,9H)；MS(ESI)276[M+Na]+。 1 H NMR (300MHz, Chloroform- d) δ6.76 (s, 1H), 4.60 (s, 2H), 3.69 (t, J = 5.7Hz, 2H), 2.56 (t, J = 5.7Hz, 2H), 2.11 (s, 3H), 1.48 (s, 9H); MS (ESI) 276 [M+Na] + .
步骤6. 2-溴-3-甲基-6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯：Step 6. 2-Bromo-3-methyl-6,7-dihydrothieno[3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000148
Figure PCTCN2016087368-appb-000148
按照制备12-3的操作方式，以3-甲基-6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯(14-12)(636mg，2.5mmol)为原料经溴代制得586mg淡黄色固体，产率为70％(石油醚：乙酸乙酯＝30：1)。According to the procedure of Preparation 12-3, 3-methyl-6,7-dihydrothieno[3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl ester (14-12) (636 mg, 2.5 mmol) was obtained by bromination of 586 mg of pale yellow solid (yield: petroleum ether: ethyl acetate = 30:1).
1H NMR(300MHz,Chloroform-d)δ4.49(s,2H),3.67(t,J＝5.6Hz,2H),2.53(t,J ＝5.5Hz,2H),2.04(s,3H),1.48(s,9H)；MS(ESI)354[M+Na]+。 1 H NMR (300MHz, Chloroform- d) δ4.49 (s, 2H), 3.67 (t, J = 5.6Hz, 2H), 2.53 (t, J = 5.5Hz, 2H), 2.04 (s, 3H), 1.48 (s, 9H); MS (ESI) 354 [M+Na] + .
步骤7. 2-甲酰基-3-甲基-6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯：Step 7. 2-Formyl-3-methyl-6,7-dihydrothieno[3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000149
Figure PCTCN2016087368-appb-000149
按照制备化合物2-3的操作方法，以2-溴-3-甲基-6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯(12-12)(586mg，1.77mmol)为原料制备得到黄色油状物379mg，产率为7％(石油醚：乙酸乙酯＝8：1)。According to the procedure for the preparation of compound 2-3, 2-bromo-3-methyl-6,7-dihydrothieno[3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl ester (12- 12) (586 mg, 1.77 mmol) was obtained as a crude material (yield: 379 g,yield
1H NMR(300MHz,Chloroform-d)δ10.02(s,1H),4.64(s,2H),3.70(t,J＝6.0Hz,2H),2.60(t,J＝5.9Hz,2H),2.42(s,3H),1.48(s,9H)；MS(ESI)304[M+Na]+。 1 H NMR (300MHz, Chloroform- d) δ10.02 (s, 1H), 4.64 (s, 2H), 3.70 (t, J = 6.0Hz, 2H), 2.60 (t, J = 5.9Hz, 2H), 2.42 (s, 3H), 1.48 (s, 9H); MS (ESI) <RTIgt ;
步骤8. 2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-3-甲基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 8. 2-(6-Fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-3-methyl-4,7-dihydrothieno[2,3 -c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000150
Figure PCTCN2016087368-appb-000150
按照制备化合物3-7的操作方法，以2-甲酰基-3-甲基-6,7-二氢噻吩并[3,2-c]吡啶-5(4H)-羧酸叔丁酯(2-12)(379mg，1.35mmol)和2,3-二氨基-5-氟苯甲酸甲酯(248mg，1.35mmol)为原料制备得到白色固体445mg，产率为74％(石油醚：乙酸乙酯＝3：1)。According to the procedure for the preparation of compound 3-7, 2-formyl-3-methyl-6,7-dihydrothieno[3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl ester (2 -12) (379 mg, 1.35 mmol) and 2,3-diamino-5-fluorobenzoic acid methyl ester (248 mg, 1.35 mmol) were obtained as a white solid 445 mg (yield: 74%) =3:1).
1H NMR(300MHz,Chloroform-d)δ10.39(brs,1H),7.66(dd,J＝8.9,2.4Hz,1H),7.58(dd,J＝9.6,2.4Hz,1H),4.66(s,2H),4.02(s,3H),3.74(t,J＝5.6Hz,2H),2.64(t,J＝5.6Hz,2H),2.52(s,3H),1.50(s,9H)；MS(ESI)446[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 10.39 (brs, 1H), 7.66 (dd, J = 8.9, 2.4 Hz, 1H), 7.58 (dd, J = 9.6, 2.4 Hz, 1H), 4.66 (s) , 2H), 4.02 (s, 3H), 3.74 (t, J = 5.6 Hz, 2H), 2.64 (t, J = 5.6 Hz, 2H), 2.52 (s, 3H), 1.50 (s, 9H); (ESI) 446 [M+H] + .
步骤9. 6-氟-2-(3-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 9. 6-Fluoro-2-(3-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole- 4-methyl formate:
Figure PCTCN2016087368-appb-000151
Figure PCTCN2016087368-appb-000151
按照制备化合物4-7的操作方法，以2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-3-甲基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(3-16)(445mg，1.00mmol)为原料去保护基得到白色粉末226mg，产率为65％(二氯甲烷：甲醇＝50：1)。According to the procedure for the preparation of compound 4-7, 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-3-methyl-4,7- Dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (3-16) (445 mg, 1.00 mmol) was used as a starting material to give a white powder 226 mg (yield: 65%) Dichloromethane: methanol = 50:1).
1H NMR(300MHz,Chloroform-d)δ10.39(brs,1H),7.65(dd,J＝8.9,2.3Hz,1H),7.57(dd,J＝9.7,2.4Hz,1H),4.07(s,2H),4.01(s,3H),3.18(t,J＝5.8Hz,2H),2.59(t,J ＝5.8Hz,2H),2.51(s,3H)；MS(ESI)346[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 10.39 (brs, 1H), 7.65 (dd, J = 8.9, 2.3 Hz, 1H), 7.57 (dd, J = 9.7, 2.4 Hz, 1H), 4.07 (s) , 2H), 4.01 (s, 3H), 3.18 (t, J = 5.8 Hz, 2H), 2.59 (t, J = 5.8 Hz, 2H), 2.51 (s, 3H); MS (ESI) 346 [M+ H] + .
步骤10. 6-氟-2-(3-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-28)：Step 10. 6-Fluoro-2-(3-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole- 4-formamide (I-28):
Figure PCTCN2016087368-appb-000152
Figure PCTCN2016087368-appb-000152
按照制备化合物I-1的操作方法，以6-氟-2-(3-甲基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(4-16)(226mg，0.65mmol)为原料氨解得到淡黄色粉末151mg，产率为70％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 6-fluoro-2-(3-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H -Benzyl[d]imidazole-4-carboxylic acid methyl ester (4-16) (226 mg, 0.65 mmol) was obtained by aminolysis to give 151 mg of pale yellow powder, yield 70% (dichloromethane:methanol = 10:1) .
1H NMR(300MHz,DMSO-d6)δ9.14(brs,1H),7.92(brs,1H),7.55(dd,J＝10.7,2.6Hz,1H),7.47(dd,J＝8.5,2.6Hz,1H),3.92(s,2H),3.17(s,2H),3.00(t,J＝5.7Hz,2H),2.46(s,3H)；13C NMR(100MHz,DMSO-d6)δ165.65,158.43(d,J＝234.4Hz,C-F),149.83,138.36,138.23,137.78,136.51,122.68,110.12(d,J＝26.1Hz,CH-CF),102.30(d,J＝27.0Hz,CH-CF),44.56,42.63,25.27,13.76；HRMS(ESI):m/z Calcd.For C16H16FN4OS[M+H]+:331.1029；Found:331.1023。 1 H NMR (300MHz, DMSO- d 6) δ9.14 (brs, 1H), 7.92 (brs, 1H), 7.55 (dd, J = 10.7,2.6Hz, 1H), 7.47 (dd, J = 8.5,2.6 Hz, 1H), 3.92 (s, 2H), 3.17 (s, 2H), 3.00 (t, J = 5.7 Hz, 2H), 2.46 (s, 3H); 13 C NMR (100 MHz, DMSO-d 6 ) δ 165 .65,158.43 (d, J=234.4 Hz, CF), 149.83, 138.36, 138.23, 137.78, 136.51, 122.68, 110.12 (d, J = 26.1 Hz, CH-CF), 102.30 (d, J = 27.0 Hz, CH- CF), 44.56, 42.63, 25.27, 13.76; HRMS (ESI): m/z Calcd. For C 16 H 16 FN 4 OS [M+H] + : 331.1029; Found: 331.1023.
实施例29. 6-氟-2-(4,5,6,7-四氢噻唑并[5,4-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-29的制备：Example 29. 6-Fluoro-2-(4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide Preparation of I-29:
步骤1. 2-氨基-6,7-二氢噻唑[4,5-c]吡啶-5(4H)-羧酸叔丁酯：Step 1. 2-Amino-6,7-dihydrothiazole [4,5-c]pyridine-5(4H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000153
Figure PCTCN2016087368-appb-000153
向溶有N-叔丁氧羰基-4-哌啶酮(22)(9.95g，50.0mmol)的环己烷(25mL)溶液中加入吡咯烷(4.35mL，52.0mmol)和一水合对甲苯磺酸(48mg，2.5mmol)，接分水器回流50分钟，冷却至室温，减压蒸干环己烷，重新溶于15mL无水甲醇，再向混合物中加入1.60g硫粉，室温搅拌20分钟，水浴冷却，缓慢向反应液中加入4.2mL的50％氰胺水溶液，水浴下继续搅拌3小时，移除水浴锅继续搅拌17小时，析出白色固体，加水40mL，过滤，水洗得8.00g白色固体，产率为63％。To a solution of N-tert-butoxycarbonyl-4-piperidone (22) (9.95 g, 50.0 mmol) in cyclohexane (25 mL) was added pyrrolidine (4.35 mL, 52.0 mmol) and p-toluene The acid (48 mg, 2.5 mmol) was refluxed with a water separator for 50 minutes, cooled to room temperature, evaporated to dryness under reduced pressure, and then redissolved in 15 mL of anhydrous methanol, and then 1.60 g of sulfur powder was added to the mixture, and stirred at room temperature for 20 minutes. Cool in a water bath, slowly add 4.2 mL of 50% aqueous cyanamide solution to the reaction solution, continue stirring for 3 hours in a water bath, remove the water bath and continue stirring for 17 hours, precipitate a white solid, add 40 mL of water, filter, and wash with water to obtain 8.00 g of white solid. The yield was 63%.
1H NMR(300MHz,DMSO-d6)δ6.78(s,2H),4.28(d,J＝2.4Hz,2H),3.55(t,J＝5.6Hz,2H),2.41(d,J＝6.3Hz,2H),1.40(s,9H)；MS(ESI)256[M+H]+。 1 H NMR (300MHz, DMSO- d 6) δ6.78 (s, 2H), 4.28 (d, J = 2.4Hz, 2H), 3.55 (t, J = 5.6Hz, 2H), 2.41 (d, J = 6.3 Hz, 2H), 1.40 (s, 9H); MS (ESI) 256 [M+H] + .
步骤2. 2-溴-6,7-二氢噻唑并[4,5-c]吡啶-5(4H)-羧酸叔丁酯：Step 2. 2-Bromo-6,7-dihydrothiazolo[4,5-c]pyridine-5(4H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000154
Figure PCTCN2016087368-appb-000154
0℃下，往2-氨基-6,7-二氢噻唑[4,5-c]吡啶-5(4H)-羧酸叔丁酯(27)(1.62g，5.0mmol)的N,N-二甲基甲酰胺(15mL)溶液中，加入溴化铜(1.70g，7.6mmol)，搅拌5分钟，在该温度下，再向反应液中缓慢滴加1.5mL亚硝酸异戊酯，滴加完毕后，缓慢升至室温继续搅拌4小时。反应液用二氯甲烷萃取(50mL×3)，有机层依次用水洗，饱和食盐水洗，保留有机层，用无水硫酸钠干燥，过滤减压浓缩，粗产品经柱层析(石油醚：乙酸乙酯＝20：1)得到淡黄色液体648mg，产率为40％。N,N- to 2-amino-6,7-dihydrothiazole [4,5-c]pyridine-5(4H)-carboxylic acid tert-butyl ester (27) (1.62 g, 5.0 mmol) at 0 °C In a solution of dimethylformamide (15 mL), copper bromide (1.70 g, 7.6 mmol) was added and stirred for 5 minutes. At this temperature, 1.5 mL of isoamyl nitrite was slowly added dropwise to the reaction solution, and the mixture was added dropwise. After completion, slowly warm to room temperature and continue to stir for 4 hours. The reaction mixture was extracted with methylene chloride (50 mL×3). The organic layer was washed with EtOAc. Ethyl ester = 20:1) 648 mg of a pale yellow liquid was obtained with a yield of 40%.
1H NMR(300MHz,Chloroform-d)δ4.56(brs,2H),3.72(t,J＝5.6Hz,2H),2.85(t,J＝5.6Hz,2H),1.48(s,9H)；MS(ESI)319[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ4.56 (brs, 2H), 3.72 (t, J = 5.6Hz, 2H), 2.85 (t, J = 5.6Hz, 2H), 1.48 (s, 9H); MS (ESI) 319 [M + H] + .
步骤3. 2-甲酰基-6,7-二氢噻唑并[4,5-c]吡啶-5(4H)-羧酸叔丁酯：Step 3. 2-Formyl-6,7-dihydrothiazolo[4,5-c]pyridine-5(4H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000155
Figure PCTCN2016087368-appb-000155
-78℃，在氮气保护下，往2-溴-6,7-二氢噻唑并[4,5-c]吡啶-5(4H)-羧酸叔丁酯(12-13)(648mg，2.0mmol)的无水四氢呋喃(30mL)溶液中缓慢滴加1.0mL正丁基锂的四氢呋喃溶液(2.5M)，继续搅拌5分钟。往反应液中加入176mg无水N,N-二甲基甲酰胺，搅拌1小时后，加入1mL水淬灭。反应液用乙酸乙酯萃取(50mL×3)，有机层依次用水、饱和食盐水洗涤，保留有机层，用无水硫酸钠干燥，过滤减压浓缩，粗产品经柱层析(石油醚：乙酸乙酯＝10：1)得无色透明油状物415mg，产率为77％。To a 2-bromo-6,7-dihydrothiazolo[4,5-c]pyridine-5(4H)-carboxylic acid tert-butyl ester (12-13) under a nitrogen atmosphere at -78 ° C (648 mg, 2.0) A solution of 1.0 mL of n-butyllithium in tetrahydrofuran (2.5 M) was slowly added dropwise to aq. After adding 176 mg of anhydrous N,N-dimethylformamide to the reaction mixture, the mixture was stirred for 1 hour and then quenched with 1 mL of water. The reaction mixture was extracted with EtOAc (EtOAc (EtOAc)EtOAc. Ethyl ester = 10:1) 415 mg of a colorless transparent oil, yield 77%.
1H NMR(300MHz,Chloroform-d)δ9.92(s,1H),4.75(s,2H),3.79(t,J＝5.8Hz,2H),2.99(t,J＝5.9Hz,2H),1.49(s,9H)；MS(ESI)269[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ9.92 (s, 1H), 4.75 (s, 2H), 3.79 (t, J = 5.8Hz, 2H), 2.99 (t, J = 5.9Hz, 2H), 1.49 (s, 9H); MS (ESI) 269 [M+H] + .
步骤4. 2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-6,7-二氢噻唑并[5,4-c]吡啶-5(4H)-羧酸叔丁酯：Step 4. 2-(6-Fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-6,7-dihydrothiazolo[5,4-c]pyridine- 5(4H)-tert-butyl carboxylic acid:
Figure PCTCN2016087368-appb-000156
Figure PCTCN2016087368-appb-000156
按照制备化合物3-7的操作方法，以2-甲酰基-6,7-二氢噻唑并[4,5-c]吡啶-5(4H)-羧酸叔丁酯(2-13)(415mg，1.55mmol)和2,3-二氨基-5-氟苯甲酸甲酯(285mg，1.55mmol)为原料制备得到白色固体228mg，产率为34％(石油醚：乙酸乙酯＝2：1)。According to the procedure for the preparation of compound 3-7, 2-formyl-6,7-dihydrothiazolo[4,5-c]pyridine-5(4H)-carboxylic acid tert-butyl ester (2-13) (415 mg) Preparation of 228 mg of a white solid in a yield of 34% ( petroleum ether: ethyl acetate = 2:1). .
1H NMR(300MHz,Chloroform-d)δ11.04(brs,1H),7.71(dd,J＝4.5,2.5Hz,1H),7.68(dd,J＝3.9,2.5Hz,1H),4.75(s,2H),4.04(s,3H),3.81(t,J＝5.5Hz,2H),2.96(t,J＝5.5Hz,2H),1.51(s,9H)；MS(ESI)433[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 11.04 (brs, 1H), 7.71 (dd, J = 4.5, 2.5 Hz, 1H), 7.68 (dd, J = 3.9, 2.5 Hz, 1H), 4.75 (s) , 2H), 4.04 (s, 3H), 3.81 (t, J = 5.5 Hz, 2H), 2.96 (t, J = 5.5 Hz, 2H), 1.51 (s, 9H); MS (ESI) 433 [M+ H] + .
步骤5. 6-氟-2-(4,5,6,7-四氢噻唑并[5,4-c]吡啶-2-基)-1H-苯并[d]咪唑-4-羧酸甲酯： Step 5. 6-Fluoro-2-(4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxylic acid A Ester:
Figure PCTCN2016087368-appb-000157
Figure PCTCN2016087368-appb-000157
按照制备化合物4-7的操作方法，以2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-6,7-二氢噻唑并[5,4-c]吡啶-5(4H)-羧酸叔丁酯(3-17)(228mg，0.53mmol)为原料去保护基得到淡棕色粉末103mg，产率为59％(二氯甲烷：甲醇＝20：1)。According to the procedure for the preparation of compound 4-7, 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-6,7-dihydrothiazolo[ 5,4-c]pyridine-5(4H)-carboxylic acid tert-butyl ester (3-17) (228 mg, 0.53 mmol) was used as a starting material to give the title compound as a pale brown powder (yield: Methanol = 20:1).
1H NMR(300MHz,Chloroform-d)δ7.71(dd,J＝4.9,2.4Hz,1H),7.68(dd,J＝3.9,2.4Hz,1H),4.17(brs,2H),4.04(s,3H),3.26(t,J＝5.8Hz,2H),2.93(t,J＝5.8Hz,2H)；MS(ESI)333[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 7.71 (dd, J = 4.9, 2.4 Hz, 1H), 7.68 (dd, J = 3.9, 2.4 Hz, 1H), 4.17 (brs, 2H), 4.04 (s) , 3H), 3.26 (t, J = 5.8 Hz, 2H), 2.93 (t, J = 5.8 Hz, 2H); MS (ESI) 333 [M+H] + .
步骤6. 6-氟-2-(4,5,6,7-四氢噻唑并[5,4-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-29)：Step 6. 6-Fluoro-2-(4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide ( I-29):
Figure PCTCN2016087368-appb-000158
Figure PCTCN2016087368-appb-000158
按照制备化合物I-1的操作方法，以6-氟-2-(4,5,6,7-四氢噻唑并[5,4-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(4-17)(103mg，0.31mmol)为原料氨解得到淡棕色粉末65mg，产率为66％(二氯甲烷：甲醇＝8：1)。According to the procedure for the preparation of compound I-1, 6-fluoro-2-(4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)-1H-benzo[d The imidazole-4-carboxylic acid methyl ester (4-17) (103 mg, 0.31 mmol) was obtained as a crude material to give a pale brown powder (yield: 66%) (yield: methylene chloride:methanol = 8:1).
1H NMR(300MHz,DMSO-d6)δ8.86(brs,1H),8.04(brs,1H),7.64(dd,J＝10.6,2.6Hz,1H),7.49(dd,J＝8.4,2.6Hz,1H),4.39(brs,2H),3.41(t,J＝6.2Hz,2H),3.06(t,J＝5.9Hz,2H)；13C NMR(101MHz,TFA-d)δ167.73,161.60(d,J＝253.6Hz,C-F),150.70,147.41,142.78,132.11(d,J＝12.8Hz,C-CH-CF),129.52,126.60,119.47(d,J＝10.8Hz,C-CH-CF),116.01,105.65(d,J＝28.4Hz,CH-CF),42.37,28.98,22.73；HRMS(ESI):m/z Calcd.For C14H13FN5OS[M+H]+:318.0825；Found:318.0820。 1 H NMR (300MHz, DMSO- d 6) δ8.86 (brs, 1H), 8.04 (brs, 1H), 7.64 (dd, J = 10.6,2.6Hz, 1H), 7.49 (dd, J = 8.4,2.6 Hz, 1H), 4.39 (brs, 2H), 3.41 (t, J = 6.2 Hz, 2H), 3.06 (t, J = 5.9 Hz, 2H); 13 C NMR (101 MHz, TFA-d) δ 167.73, 161.60 ( d, J = 253.6 Hz, CF), 150.70, 147.41, 142.78, 132.11 (d, J = 12.8 Hz, C-CH-CF), 129.52, 126.60, 119.47 (d, J = 10.8 Hz, C-CH-CF ), 116.01, 105.65 (d, J = 28.4 Hz, CH-CF), 42.37, 28.98, 22.73; HRMS (ESI): m/z Calcd. For C 14 H 13 FN 5 OS [M+H] + : 318.0825 Found: 318.0820.
实施例30. 6-氟-1-甲基-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-30的制备：Example 30. 6-Fluoro-1-methyl-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole Preparation of 4-carboxamide, I-30:
步骤1. 2-(6-氟-4-(甲氧基羰基)-1-甲基-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 1. 2-(6-Fluoro-4-(methoxycarbonyl)-1-methyl-1H-benzo[d]imidazol-2-yl)-4,7-dihydrothieno[2,3 -c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000159
Figure PCTCN2016087368-appb-000159
往2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(500mg，1.16mmol)的丙酮(5mL)溶液中，加入K2CO3(240mg，1.74mmol)和碘甲烷(198mg，1.39mmol)，搅拌过夜，反应液用乙酸乙酯萃取(20mL×3)，有机层依次用水洗，饱和食盐水洗，保留有机层，用无水硫酸钠干燥，过滤减压浓缩，粗产品经柱层析(石油醚：乙酸乙酯＝3：1)得淡黄色固体222mg，产率为43％。To 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7-dihydrothieno[2,3-c]pyridine-6 ( 5H) - carboxylate (500mg, 1.16mmol) in acetone (5mL) was added K 2 CO 3 (240mg, 1.74mmol ) and iodomethane (198mg, 1.39mmol), stirred overnight, the reaction solution was neutralized with acetic acid The ethyl ester was extracted (20 mL×3), and the organic layer was washed with water and brine, and then evaporated. : 1) 222 mg of a pale yellow solid, yield 43%.
1H NMR(300MHz,Chloroform-d)δ7.72(dd,J＝10.2,2.4Hz,1H),7.37(s,1H),7.24(dd,J＝9.8,2.4Hz,1H),4.69(s,2H),4.04(s,3H),3.96(s,3H),3.71(t,J＝5.8Hz,2H),2.77(t,J＝6.0Hz,2H),1.50(s,9H)；MS(ESI)446[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ7.72 (dd, J = 10.2,2.4Hz, 1H), 7.37 (s, 1H), 7.24 (dd, J = 9.8,2.4Hz, 1H), 4.69 (s , 2H), 4.04 (s, 3H), 3.96 (s, 3H), 3.71 (t, J = 5.8 Hz, 2H), 2.77 (t, J = 6.0 Hz, 2H), 1.50 (s, 9H); (ESI) 446 [M+H] + .
步骤2. 6-氟-1-甲基-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 2. 6-Fluoro-1-methyl-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole- 4-methyl formate:
Figure PCTCN2016087368-appb-000160
Figure PCTCN2016087368-appb-000160
按照制备化合物4-7的操作方法，以2-(6-氟-4-(甲氧基羰基)-1-甲基-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(222mg，0.50mmol)为原料去保护基得到白色粉末154mg，产率为89％(二氯甲烷：甲醇＝20：1)。According to the procedure for the preparation of compound 4-7, 2-(6-fluoro-4-(methoxycarbonyl)-1-methyl-1H-benzo[d]imidazol-2-yl)-4,7- Dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (222 mg, 0.50 mmol) was used as a starting material to give a white powder 154 mg (yield: 89%) =20:1).
1H NMR(300MHz,Chloroform-d)δ7.69(dd,J＝10.2,2.5Hz,1H),7.35(s,1H),7.20(dd,J＝8.0,2.5Hz,1H),4.09(brs,2H),4.03(s,3H),3.94(s,3H),3.16(t,J＝5.8Hz,2H),2.71(t,J＝5.8Hz,2H),1.24(s,9H)；MS(ESI)346[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ7.69 (dd, J = 10.2,2.5Hz, 1H), 7.35 (s, 1H), 7.20 (dd, J = 8.0,2.5Hz, 1H), 4.09 (brs , 2H), 4.03 (s, 3H), 3.94 (s, 3H), 3.16 (t, J = 5.8 Hz, 2H), 2.71 (t, J = 5.8 Hz, 2H), 1.24 (s, 9H); (ESI) 346 [M+H] + .
步骤3. 6-氟-1-甲基-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-30)：Step 3. 6-Fluoro-1-methyl-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole- 4-formamide (I-30):
Figure PCTCN2016087368-appb-000161
Figure PCTCN2016087368-appb-000161
按照制备化合物I-1的操作方法，以6-氟-1-甲基-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(154mg，0.45mmol)为原料氨解得到白色粉末116mg，产率为79％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 6-fluoro-1-methyl-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H Methyl benzo[d]imidazole-4-carboxylate (154 mg, 0.45 mmol) was obtained as a white powder to afford white powder (yield: &lt;RTIgt;
1H NMR(500MHz,DMSO-d6)δ9.05(brs,1H),8.00(brs,1H),7.81(d,J＝8.6Hz,1H),7.60(s,1H),7.58(d,J＝12.9Hz,1H),4.01(brs,5H),3.03(brs,2H),2.68(brs,2H)；13C NMR(125MHz,DMSO-d6)δ165.22,158.77(d,J＝237.0Hz,C-F),149.48,138.10(d,J＝12.9Hz,C-CH-CF),136.97,136.02,130.17,127.91,123.36(d,J＝8.6Hz,C-CH-CF),110.93(d,J＝26.3Hz,CH-CF),101.49(d,J＝28.6Hz,CH-CF),44.28,42.66,32.80,26.01； HRMS(ESI):m/z Calcd.For C16H16FN4OS[M]+:331.1029；Found:331.1024。 1 H NMR (500MHz, DMSO- d 6) δ9.05 (brs, 1H), 8.00 (brs, 1H), 7.81 (d, J = 8.6Hz, 1H), 7.60 (s, 1H), 7.58 (d, J = 12.9 Hz, 1H), 4.01 (brs, 5H), 3.03 (brs, 2H), 2.68 (brs, 2H); 13 C NMR (125 MHz, DMSO-d 6 ) δ 165.22, 158.77 (d, J = 237.0 Hz) , CF), 149.48, 138.10 (d, J = 12.9 Hz, C-CH-CF), 136.97, 136.02, 130.17, 127.91, 123.36 (d, J = 8.6 Hz, C-CH-CF), 110.93 (d, J=26.3 Hz, CH-CF), 101.49 (d, J=28.6 Hz, CH-CF), 44.28, 42.66, 32.80, 26.01; HRMS (ESI): m/z Calcd.For C 16 H 16 FN 4 OS [M] + : 331.1029; Found: 331.1024.
I-30盐酸盐的制备方案：Preparation scheme of I-30 hydrochloride:
将游离态的I-30(100mg)溶于二氯甲烷(100ml)和甲醇(10ml)混合溶剂中，冰浴下随后缓慢充30分钟HCl(g),随后与室温下搅拌过夜，浓缩除去大部分溶剂，过滤，干燥得到I-30盐酸盐。The free I-30 (100 mg) was dissolved in a mixed solvent of dichloromethane (100 ml) and methanol (10 ml), and then slowly stirred for 30 minutes HCl (g) in an ice bath, then stirred at room temperature overnight, concentrated to remove most Solvent, filter and dry to give the I-30 hydrochloride.
实施例31. 1-乙基-6-氟-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺，I-31的制备：Example 31. 1-Ethyl-6-fluoro-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole Preparation of 4-carboxamide, I-31:
步骤1. 2-(1-乙基-6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 1. 2-(1-Ethyl-6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7-dihydrothieno[2,3 -c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000162
Figure PCTCN2016087368-appb-000162
往2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(500mg，1.16mmol)的N,N-二甲基甲酰胺(5mL)溶液中，加入Cs2CO3(567mg，1.74mmol)和溴乙烷(253mg，2.32mmol)，90℃反应过夜，反应液用乙酸乙酯萃取(30mL×3)，有机层依次用水、饱和食盐水洗涤，保留有机层，用无水硫酸钠干燥，过滤减压浓缩，粗产品经柱层析(石油醚：乙酸乙酯＝3：1)得淡黄色固体245mg，产率为46％。To 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7-dihydrothieno[2,3-c]pyridine-6 ( 5H) - carboxylate (500mg, 1.16mmol) in N, N- dimethylformamide (5mL) was added Cs 2 CO 3 (567mg, 1.74mmol ) and bromoethane (253mg, 2.32mmol The reaction was carried out at 90 ° C overnight, and the mixture was extracted with ethyl acetate (30 mL×3). Chromatography (petroleum ether: ethyl acetate = 3:1) afforded 245 mg of pale yellow solid.
1H NMR(300MHz,Chloroform-d)δ7.71(dd,J＝10.4,2.0Hz,1H),7.37(s,1H),7.24(dd,J＝10.1,2.4Hz,1H),4.68(s,2H),4.41(d,J＝7.2Hz,2H),4.04(s,3H),3.72(t,J＝5.1Hz,2H),2.75(t,J＝5.1Hz,2H),1.50(s,9H)；MS(ESI)460[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ7.71 (dd, J = 10.4,2.0Hz, 1H), 7.37 (s, 1H), 7.24 (dd, J = 10.1,2.4Hz, 1H), 4.68 (s , 2H), 4.41 (d, J = 7.2 Hz, 2H), 4.04 (s, 3H), 3.72 (t, J = 5.1 Hz, 2H), 2.75 (t, J = 5.1 Hz, 2H), 1.50 (s , 9H); MS (ESI) 460 [M+H] + .
步骤2. 1-乙基-6-氟-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 2. 1-Ethyl-6-fluoro-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole- 4-methyl formate:
Figure PCTCN2016087368-appb-000163
Figure PCTCN2016087368-appb-000163
按照制备化合物4-7的操作方法，以2-(1-乙基-6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(6-2)(245mg，0.53mmol)为原料去保护基得到棕色固体156mg，产率为82％(二氯甲烷：甲醇＝30：1)。According to the procedure for the preparation of compound 4-7, 2-(1-ethyl-6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7- Dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (6-2) (245 mg, 0.53 mmol) was used as the starting material to afford the title compound to afford 156 mg as a brown solid. Dichloromethane: methanol = 30:1).
1H NMR(300MHz,Chloroform-d)δ7.71(dd,J＝10.2,2.4Hz,1H),7.36(d,J＝2.0Hz,1H),7.24(dd,J＝9.1,3.1Hz,1H),4.40(q,J＝7.3Hz,2H),4.12(s,2H),4.04(s,3H),3.20(t,J＝5.8Hz,2H),2.75(t,J＝5.6Hz,2H),1.49(t,J＝7.3Hz,3H)；MS(ESI)360[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 7.71 (dd, J = 10.2, 2.4 Hz, 1H), 7.36 (d, J = 2.0 Hz, 1H), 7.24 (dd, J = 9.1, 3.1 Hz, 1H) ), 4.40 (q, J = 7.3 Hz, 2H), 4.12 (s, 2H), 4.04 (s, 3H), 3.20 (t, J = 5.8 Hz, 2H), 2.75 (t, J = 5.6 Hz, 2H) ), 1.49 (t, J = 7.3 Hz, 3H); MS (ESI) 360 [M+H] + .
步骤3. 1-乙基-6-氟-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-31)：Step 3. 1-Ethyl-6-fluoro-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole- 4-formamide (I-31):
Figure PCTCN2016087368-appb-000164
Figure PCTCN2016087368-appb-000164
按照制备化合物I-1的操作方法，以1-乙基-6-氟-2-(4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(156mg，0.43mmol)为原料氨解得到白色粉末120mg，产率为81％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 1-ethyl-6-fluoro-2-(4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1H - Benzo[d]imidazole-4-carboxylic acid methyl ester (156 mg, 0.43 mmol) was obtained as a crude material to give a white powder (yield: 120%) (yield: methylene chloride:methanol = 10:1).
1H NMR(500MHz,DMSO-d6)δ9.04(brs,1H),8.03(brs,1H),7.88(dd,J＝8.9,2.6Hz,1H),7.60(dd,J＝9.2,2.4Hz,1H),7.59(s,1H),4.54(q,J＝7.2Hz,2H),4.10(s,2H),3.11(t,J＝5.7Hz,2H),2.76(t,J＝5.7Hz,2H),1.40(t,J＝7.1Hz,3H)；13C NMR(125MHz,DMSO-d6)δ165.11,158.87(d,J＝237.1Hz,C-F),148.49,137.47,137.08(d,J＝8.8Hz,C-CH-CF),137.07,135.83,129.43,128.15,123.60(d,J＝8.4Hz,C-CH-CF),111.16(d,J＝26.6Hz,CH-CF),101.56(d,J＝28.2Hz,CH-CF),43.70,42.21,29.44,25.22,15.17；HRMS(ESI):m/z Calcd.For C17H18FN4OS[M+H]+:345.1185；Found:345.1177。 1 H NMR (500 MHz, DMSO-d 6 ) δ 9.04 (brs, 1H), 8.03 (brs, 1H), 7.78 (dd, J = 8.9, 2.6 Hz, 1H), 7.60 (dd, J = 9.2, 2.4 Hz, 1H), 7.59 (s, 1H), 4.54 (q, J = 7.2 Hz, 2H), 4.10 (s, 2H), 3.11 (t, J = 5.7 Hz, 2H), 2.76 (t, J = 5.7) Hz, 2H), 1.40 (t, J = 7.1 Hz, 3H); 13 C NMR (125MHz, DMSO-d 6 ) δ 165.11, 158.87 (d, J = 237.1 Hz, CF), 148.49, 137.47, 137.08 (d, J = 8.8 Hz, C-CH-CF), 137.07, 135.83, 129.43, 128.15, 123.60 (d, J = 8.4 Hz, C-CH-CF), 111.16 (d, J = 26.6 Hz, CH-CF), 101.56 (d, J = 28.2 Hz, CH-CF), 43.70, 42.21, 29.44, 25.22, 15.17; HRMS (ESI): m/z Calcd. For C 17 H 18 FN 4 OS [M+H] + :345.1185 Found: 345.1177.
实施例32. 6-氟-2-(7-异丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1-甲基-1H-苯并[d]咪唑-4-甲酰胺，I-32的制备：Example 32. 6-Fluoro-2-(7-isobutyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1-methyl-1H- Preparation of benzo[d]imidazole-4-carboxamide, I-32:
步骤1. 2-(6-氟-4-(甲氧基羰基)-1-甲基-1H-苯并[d]咪唑-2-基)-7-异丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 1. 2-(6-Fluoro-4-(methoxycarbonyl)-1-methyl-1H-benzo[d]imidazol-2-yl)-7-isobutyl-4,7-dihydro Thio[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000165
Figure PCTCN2016087368-appb-000165
按照实施例30中步骤1的操作方式，以2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-7-异丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(386mg，0.79mmol)为原料制备得到白色固体263mg，产率为66％(石油醚：乙酸乙酯＝8：1)。Following the procedure of Step 1 in Example 30, 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-7-isobutyl-4, Preparation of 7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (386 mg, 0.79 mmol) as a white solid 263 mg (yield: 66%) Ester = 8:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ7.71(dd,J＝10.2,2.5Hz,1H),7.30(s,1H),7.23(dd,J＝7.9,2.5Hz,1H),5.53–5.42(m,1H),4.26–4.12(m,1H),4.04(s,3H),3.96(s,3H),3.19–2.96(m,2H),2.63–2.55(m,1H),1.96–1.73(m,1H),1.63–1.54(m,2H),1.49(s,9H),1.05(d,J＝6.3Hz,3H),0.99(d,J＝6.5Hz,3H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ7.71(dd,J＝10.2,2.5Hz,1H),7.30(s,1H),7.23(dd,J＝7.9,2.5Hz,1H),5.34–5.21(m,1H),4.47–4.33(m,1H),4.04(s,3H),3.96(s,3H),2.93–2.69(m,2H),2.69–2.62(m,1H),2.63–2.55(m,1H),1.96–1.73(m,1H),1.63–1.54 (m,2H),1.49(s,9H),1.05(d,J＝6.3Hz,3H),0.99(d,J＝6.5Hz,3H)；MS(ESI)502[M+H]+。Conformational isomer 1: 1 H NMR (300 MHz, Chloroform-d) δ 7.71 (dd, J = 10.2, 2.5 Hz, 1H), 7.30 (s, 1H), 7.23 (dd, J = 7.9, 2.5 Hz, 1H), 5.53–5.42 (m, 1H), 4.26–4.12 (m, 1H), 4.04 (s, 3H), 3.96 (s, 3H), 3.19–2.96 (m, 2H), 2.63–2.55 (m, 1H), 1.96–1.73 (m, 1H), 1.63–1.54 (m, 2H), 1.49 (s, 9H), 1.05 (d, J = 6.3 Hz, 3H), 0.99 (d, J = 6.5 Hz, 3H) Conformation isomer 2: 1 H NMR (300 MHz, Chloroform-d) δ 7.71 (dd, J = 10.2, 2.5 Hz, 1H), 7.30 (s, 1H), 7.23 (dd, J = 7.9, 2.5) Hz, 1H), 5.34–5.21 (m, 1H), 4.47–4.33 (m, 1H), 4.04 (s, 3H), 3.96 (s, 3H), 2.93–2.69 (m, 2H), 2.69–2.62 ( m, 1H), 2.63–2.55 (m, 1H), 1.96–1.73 (m, 1H), 1.63–1.54 (m, 2H), 1.49 (s, 9H), 1.05 (d, J = 6.3 Hz, 3H) , 0.99 (d, J = 6.5 Hz, 3H); MS (ESI) 502 [M+H] + .
步骤2. 6-氟-2-(7-异丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1-甲基-1H-苯并[d]咪唑-4-甲酸甲酯：Step 2. 6-Fluoro-2-(7-isobutyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1-methyl-1H-benzene And [d]imidazole-4-carboxylic acid methyl ester:
Figure PCTCN2016087368-appb-000166
Figure PCTCN2016087368-appb-000166
按照制备化合物4-7的操作方法，以2-(6-氟-4-(甲氧基羰基)-1-甲基-1H-苯并[d]咪唑-2-基)-7-异丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(263mg，0.52mmol)为原料去保护基得到白色固体192mg，产率为91％(二氯甲烷：甲醇＝50：1)。According to the procedure for the preparation of compound 4-7, 2-(6-fluoro-4-(methoxycarbonyl)-1-methyl-1H-benzo[d]imidazol-2-yl)-7-isobutyl Benzyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (263 mg, 0.52 mmol) was obtained as a starting material to give a white solid. (Dichloromethane: methanol = 50:1).
1H NMR(400MHz,DMSO-d6)δ7.72(dd,J＝8.8,2.6Hz,1H),7.36(s,1H),7.21(d,J＝10.4,2.5Hz,1H),4.01(s,3H),3.20(brs,1H),2.83(brs,1H),2.61(brs,2H),1.95(s,1H),1.66–1.55(m,1H),1.54-1.48(m,1H),0.99(d,J＝6.1Hz,3H),0.97(d,J＝6.1Hz,3H)；MS(ESI)402[M+H]+。 1 H NMR (400 MHz, DMSO-d 6 ) δ 7.72 (dd, J = 8.8, 2.6 Hz, 1H), 7.36 (s, 1H), 7.21. (d, J = 10.4, 2.5 Hz, 1H), 4.01 ( s, 3H), 3.20 (brs, 1H), 2.83 (brs, 1H), 2.61 (brs, 2H), 1.95 (s, 1H), 1.66 - 1.55 (m, 1H), 1.54-1.48 (m, 1H) , 0.99 (d, J = 6.1 Hz, 3H), 0.97 (d, J = 6.1 Hz, 3H); MS (ESI) 402 [M+H] + .
步骤3. 6-氟-2-(7-异丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1-甲基-1H-苯并[d]咪唑-4-甲酰胺(I-32)：Step 3. 6-Fluoro-2-(7-isobutyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-1-methyl-1H-benzene And [d]imidazole-4-carboxamide (I-32):
Figure PCTCN2016087368-appb-000167
Figure PCTCN2016087368-appb-000167
按照制备化合物I-1的操作方法，以6-氟-2-(7-异丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-1-甲基-1H-苯并[d]咪唑-4-甲酸甲酯(192mg，0.48mmol)为原料氨解得到白色粉末128mg，产率为69％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 6-fluoro-2-(7-isobutyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)- The methyl ester of 1-methyl-1H-benzo[d]imidazole-4-carboxylate (192 mg, 0.48 mmol) was obtained as a white powder to give a white powder (yield: 69 mg, yield: 69% (dichloromethane:methanol = 10:1) .
1H NMR(400MHz,DMSO-d6)δ9.04(brs,1H),7.96(brs,1H),7.79(dd,J＝8.8,2.6Hz,1H),7.56(s,1H),7.55(d,J＝10.4,2.5Hz,1H),4.00(s,3H),3.13(brs,1H),2.81(brs,1H),2.59(brs,2H),1.94(s,1H),1.66–1.54(m,1H),1.53-1.48(m,1H),0.94(d,J＝6.1Hz,3H),0.92(d,J＝6.1Hz,3H)；13C NMR(100MHz,DMSO-d6)δ165.15,158.70(d,J＝236.9Hz,C-F),149.56,138.07(d,J＝13.4Hz,C-CH-CF),136.97,136.64,130.38,127.38,123.30(d,J＝8.5Hz,C-CH-CF),110.84(d,J＝26.3Hz,CH-CF),101.43(d,J＝28.2Hz,CH-CF),52.48,47.06,32.77,24.59,24.04,21.86；HRMS(ESI):m/z Calcd.For C20H24FN4OS[M+H]+:387.1655；Found:387.1649。 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.04 (brs, 1H), 7.96 (brs, 1H), 7.79 (dd, J = 8.8, 2.6 Hz, 1H), 7.56 (s, 1H), 7.55 ( d, J = 10.4, 2.5 Hz, 1H), 4.00 (s, 3H), 3.13 (brs, 1H), 2.81 (brs, 1H), 2.59 (brs, 2H), 1.94 (s, 1H), 1.66 - 1.54 (m, 1H), 1.53-1.48 (m, 1H), 0.94 (d, J = 6.1 Hz, 3H), 0.92 (d, J = 6.1 Hz, 3H); 13 C NMR (100 MHz, DMSO-d 6 ) δ 165.15, 158.70 (d, J = 236.9 Hz, CF), 149.56, 138.07 (d, J = 13.4 Hz, C-CH-CF), 136.97, 136.64, 130.38, 127.38, 123.30 (d, J = 8.5 Hz, C -CH-CF), 110.84 (d, J = 26.3 Hz, CH-CF), 101.43 (d, J = 28.2 Hz, CH-CF), 52.48, 47.06, 32.77, 24.59, 24.04, 21.86; HRMS (ESI) : m/z Calcd. For C 20 H 24 FN 4 OS [M+H] + : 387.1655; Found: 387.1649.
实施例33. 2-(7-叔丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1-甲基-1H-苯并[d]咪唑-4-甲酰胺，I-33的制备： Example 33. 2-(7-tert-Butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1-methyl-1H- Preparation of benzo[d]imidazole-4-carboxamide, I-33:
步骤1. 7-叔丁基-2-(6-氟-4-(甲氧基羰基)-1-甲基-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 1. 7-tert-Butyl-2-(6-fluoro-4-(methoxycarbonyl)-1-methyl-1H-benzo[d]imidazol-2-yl)-4,7-dihydro Thio[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000168
Figure PCTCN2016087368-appb-000168
按照实施例30中步骤1的操作方式，以2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-7-异丁基-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(459mg，0.94mmol)为原料制备得到白色固体292mg，产率为62％(石油醚：乙酸乙酯＝8：1)。Following the procedure of Step 1 in Example 30, 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-7-isobutyl-4, 7-Dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (459 mg, 0.94 mmol) was obtained as a starting material to give 292 mg of white solid (yield: 62%). Ester = 8:1).
构象异构体1：1H NMR(300MHz,Chloroform-d)δ7.72(dd,J＝10.3,2.5Hz,1H),7.37(dd,J＝11.5,2.5Hz,1H),7.23(s,1H),5.22(brs,1H),4.30(dd,J＝13.5,5.6Hz,1H),4.04(s,3H),3.97(s,3H),3.40–3.14(m,2H),2.66–2.57(m,1H),1.48(s,9H),1.12(s,9H)；构象异构体2：1H NMR(300MHz,Chloroform-d)δ7.72(dd,J＝10.3,2.5Hz,1H),7.37(d,J＝11.5Hz,1H),7.23(s,1H),5.02(brs,1H),4.51(dd,J＝13.7,6.1Hz,1H),4.04(s,3H),3.98(s,3H),2.71–2.64(m,1H),2.66–2.57(m,1H),1.48(s,9H),1.12(s,9H)；MS(ESI)502[M+H]+。Conformational isomer 1: 1 H NMR (300 MHz, Chloroform-d) δ 7.72 (dd, J = 10.3, 2.5 Hz, 1H), 7.37 (dd, J = 11.5, 2.5 Hz, 1H), 7.23 (s, 1H), 5.22 (brs, 1H), 4.30 (dd, J = 13.5, 5.6 Hz, 1H), 4.04 (s, 3H), 3.97 (s, 3H), 3.40 - 3.14 (m, 2H), 2.66 - 2.57 (m, 1H), 1.48 (s, 9H), 1.12 (s, 9H); conformational isomer 2: 1 H NMR (300 MHz, Chloroform-d) δ 7.72 (dd, J = 10.3, 2.5 Hz, 1H) ), 7.37 (d, J = 11.5 Hz, 1H), 7.23 (s, 1H), 5.02 (brs, 1H), 4.51 (dd, J = 13.7, 6.1 Hz, 1H), 4.04 (s, 3H), 3.98 (s, 3H), 2.71 - 2.64 (m, 1H), 2.66 - 2.57 (m, 1H), 1.48 (s, 9H), 1.12 (s, 9H); MS (ESI) 502 [M+H] + .
步骤2. 2-(7-叔丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1-甲基-1H-苯并[d]咪唑-4-甲酸甲酯：Step 2. 2-(7-tert-Butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1-methyl-1H-benzene And [d]imidazole-4-carboxylic acid methyl ester:
Figure PCTCN2016087368-appb-000169
Figure PCTCN2016087368-appb-000169
按照制备化合物4-7的操作方法，以7-叔丁基-2-(6-氟-4-(甲氧基羰基)-1-甲基-1H-苯并[d]咪唑-2-基)-4,7-二氢噻吩并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(292mg，0.58mmol)为原料去保护基得到白色固体196mg，产率为84％(二氯甲烷：甲醇＝50：1)。According to the procedure for the preparation of compound 4-7, 7-tert-butyl-2-(6-fluoro-4-(methoxycarbonyl)-1-methyl-1H-benzo[d]imidazol-2-yl 4,7-Dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (292 mg, 0.58 mmol) was used as a starting material to give the title compound as a white solid. (Dichloromethane: methanol = 50:1).
1H NMR(300MHz,Chloroform-d)δ7.72(dd,J＝10.2,2.4Hz,1H),7.35(s,1H),7.23(dd,J＝7.95,2.4Hz,1H),4.05(s,3H),3.98(s,3H),3.86(s,1H),3.37(ddd,J＝12.4,4.9,2.8Hz,1H),2.91(ddd,J＝12.3,9.5,5.3Hz,1H),2.75-2.66(m,2H),1.14(s,9H)；MS(ESI)402[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 7.72 (dd, J = 10.2, 2.4 Hz, 1H), 7.35 (s, 1H), 7.23 (dd, J = 7.95, 2.4 Hz, 1H), 4.05 (s) , 3H), 3.98 (s, 3H), 3.86 (s, 1H), 3.37 (ddd, J = 12.4, 4.9, 2.8 Hz, 1H), 2.91 (ddd, J = 12.3, 9.5, 5.3 Hz, 1H), 2.75-2.66 (m, 2H), 1.14 (s, 9H); MS (ESI) 402 [M+H] + .
步骤3. 2-(7-叔丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1-甲基-1H-苯并[d]咪唑-4-甲酰胺(I-33)： Step 3. 2-(7-tert-Butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro-1-methyl-1H-benzene And [d]imidazole-4-carboxamide (I-33):
Figure PCTCN2016087368-appb-000170
Figure PCTCN2016087368-appb-000170
按照制备化合物I-1的操作方法，以2-(7-叔丁基-4,5,6,7-四氢噻吩并[2,3-c]吡啶-2-基)-6-氟-1-甲基-1H-苯并[d]咪唑-4-甲酸甲酯(196mg，0.49mmol)为原料氨解得到白色粉末132mg，产率为70％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 2-(7-tert-butyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-6-fluoro- Methyl 1-methyl-1H-benzo[d]imidazole-4-carboxylate (196 mg, 0.49 mmol) was obtained as a crude material to give a white powder (132 mg, yield: 70% (dichloromethane:methanol = 10:1) .
1H NMR(400MHz,TFA-d)δ8.20–7.52(m,3H),4.74(brs,1H),4.18(s,3H),4.06(brs,1H),3.57(brs,1H),3.21(brs,2H),1.32(s,9H)；13C NMR(101MHz,TFA-d)δ167.72,159.74,146.42,137.47,136.93,135.38,134.55,125.74,119.76,118.95,104.22,103.92,65.46,42.79,35.06,32.31,24.56,21.62；HRMS(ESI):m/z Calcd.For C20H24FN4OS[M+H]+:387.1655；Found:387.1648。 1 H NMR (400MHz, TFA- d) δ8.20-7.52 (m, 3H), 4.74 (brs, 1H), 4.18 (s, 3H), 4.06 (brs, 1H), 3.57 (brs, 1H), 3.21 (brs, 2H), 1.32 (s, 9H); 13 C NMR (101MHz, TFA-d) δ 167.72, 159.74, 146.42, 137.47, 136.93, 135.38, 134.55, 125.74, 119.76, 118.95, 104.22, 103.92, 65.46, 42.79 , 35.06, 32.31, 24.56, 21.62; HRMS (ESI): m/z Calcd. For C 20 H 24 FN 4 OS [M+H] + : 387.1655; Found: 387.1648.
实施例34. 6-氟-2-(4,5,6,7-四氢呋喃并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-34)：Example 34. 6-Fluoro-2-(4,5,6,7-tetrahydrofuro[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide (I -34):
步骤1. 4,7-二氢呋喃并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 1. 4,7-Dihydrofuro[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000171
Figure PCTCN2016087368-appb-000171
参照文献制备(参考文献：Haginoya,N.；Kobayashi,S.；Komoriya,S.；Yoshino,T.；Suzuki,M.；Shimada,T.；Watanabe,K.；Hirokawa,Y.；Furugori,T.and Nagahara,T.Synthesis and Conformational Analysis of a Non-Amidine Factor Xa Inhibitor That Incorporates 5-Methyl-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridine as S4Binding Element.J.Med.Chem.2004,47,5167-5182.)。Reference preparation (References: Haginoya, N.; Kobayashi, S.; Komoriya, S.; Yoshino, T.; Suzuki, M.; Shimada, T.; Watanabe, K.; Hirokawa, Y.; Furugori, T .and Nagahara, T.Synthesis and Conformational Analysis of a Non-Amidine Factor Xa Inhibitor That Incorporates 5-Methyl-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridine as S4Binding Element.J.Med.Chem .2004, 47, 5167-5182.).
1H NMR(300MHz,Chloroform-d)δ7.39(d,J＝1.6Hz,1H),6.24(d,J＝1.6Hz,1H),4.44(s,2H),3.63(t,J＝5.3Hz,2H),2.51(t,J＝5.3Hz,2H),1.48(s,9H)；MS(ESI)246[M+Na]+。 1 H NMR (300MHz, Chloroform- d) δ7.39 (d, J = 1.6Hz, 1H), 6.24 (d, J = 1.6Hz, 1H), 4.44 (s, 2H), 3.63 (t, J = 5.3 Hz, 2H), 2.51 (t, J = 5.3 Hz, 2H), 1.48 (s, 9H); MS (ESI) 246 [M+Na] + .
步骤2. 2-甲酰基-4,7-二氢呋喃并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 2. 2-Formyl-4,7-dihydrofuro[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000172
Figure PCTCN2016087368-appb-000172
按照制备化合物2-3的操作方法，以4,7-二氢呋喃并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(14-13)(165mg，0.74mmol)为原料制备得到浅黄色液体33mg，产率为18％(石油醚：乙酸乙酯＝10：1)。According to the procedure for the preparation of compound 2-3, 4,7-dihydrofuro[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (14-13) (165 mg, 0.74 mmol) was used. The starting material was obtained to give a pale yellow liquid (yield: 33 mg) (yield: petroleum ether: ethyl acetate = 10:1).
1H NMR(300MHz,Chloroform-d)δ9.55(s,1H),7.09(s,1H),4.54(s,2H),3.67(t, J＝5.2Hz,2H),2.60(t,J＝5.2Hz,2H),1.48(s,9H)；MS(ESI)274[M+Na]+。 1 H NMR (300 MHz, Chloroform-d) δ 9.55 (s, 1H), 7.09 (s, 1H), 4.54 (s, 2H), 3.67 (t, J = 5.2 Hz, 2H), 2.60 (t, J) = 5.2 Hz, 2H), 1.48 (s, 9H); MS (ESI) 274 [M+Na] + .
步骤3. 2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢呋喃并[2,3-c]吡啶-6(5H)-羧酸叔丁酯：Step 3. 2-(6-Fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7-dihydrofuro[2,3-c]pyridine- 6(5H)-tert-butyl carboxylic acid:
Figure PCTCN2016087368-appb-000173
Figure PCTCN2016087368-appb-000173
按照制备化合物3-7的操作方法，以2-甲酰基-4,7-二氢呋喃并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(2-14)(33mg，0.13mmol)和2,3-二氨基-5-氟苯甲酸甲酯(24mg，0.13mmol)为原料制备得到淡黄色固体46mg，产率为86％(石油醚：乙酸乙酯＝3：1)。According to the procedure for the preparation of compound 3-7, 2-formyl-4,7-dihydrofuro[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (2-14) (33 mg) , 0.13 mmol) and 2,3-diamino-5-fluorobenzoic acid methyl ester (24 mg, 0.13 mmol) as a starting material to give 46 mg of pale yellow solid (yield: 86%) ( petroleum ether: ethyl acetate = 3:1) ).
1H NMR(300MHz,Chloroform-d)δ7.82(dd,J＝8.0,1.9Hz,1H),7.69(dd,J＝9.1,1.9Hz,1H),7.56-7.46(m,2H),4.57(s,2H),4.06(s,3H),3.768t,J＝5.8Hz,2H),2.63(t,J＝5.8Hz,2H),1.51(s,9H)；MS(ESI)416[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 7.82 (dd, J = 8.0, 1.9 Hz, 1H), 7.69 (dd, J = 9.1, 1.9 Hz, 1H), 7.56-7.46 (m, 2H), 4.57 (s, 2H), 4.06 (s, 3H), 3.768t, J = 5.8 Hz, 2H), 2.63 (t, J = 5.8 Hz, 2H), 1.51 (s, 9H); MS (ESI) 416 [M +H] + .
步骤4. 6-氟-2-(4,5,6,7-四氢呋喃并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯：Step 4. 6-Fluoro-2-(4,5,6,7-tetrahydrofuro[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxylic acid methyl ester:
Figure PCTCN2016087368-appb-000174
Figure PCTCN2016087368-appb-000174
按照制备化合物4-的操作方法，以2-(6-氟-4-(甲氧基羰基)-1H-苯并[d]咪唑-2-基)-4,7-二氢呋喃并[2,3-c]吡啶-6(5H)-羧酸叔丁酯(3-18)(46mg，0.11mmol)为原料去保护基得到白色固体28mg，产率为80％(二氯甲烷：甲醇＝50：1)。According to the procedure for the preparation of compound 4-, 2-(6-fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-4,7-dihydrofuran[2] , 3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (3-18) (46 mg, 0.11 mmol) was obtained as a starting material to give the title compound as a white solid (yield: 28 mg, m. 50:1).
1H NMR(300MHz,Chloroform-d)δ10.50(brs,1H),7.61(dd,J＝7.6,2.4Hz,1H),7.59((dd,J＝8.4,2.4Hz,1H),7.08(s,1H),4.02(s,3H),3.98(s,2H),3.10(t,J＝5.6Hz,2H),2.59(t,J＝5.3Hz,2H)；MS(ESI)316[M+H]+。 1 H NMR (300MHz, Chloroform- d) δ10.50 (brs, 1H), 7.61 (dd, J = 7.6,2.4Hz, 1H), 7.59 ((dd, J = 8.4,2.4Hz, 1H), 7.08 ( s, 1H), 4.02 (s, 3H), 3.98 (s, 2H), 3.10 (t, J = 5.6 Hz, 2H), 2.59 (t, J = 5.3 Hz, 2H); MS (ESI) 316 [M +H] + .
步骤5. 6-氟-2-(4,5,6,7-四氢呋喃并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-34)：Step 5. 6-Fluoro-2-(4,5,6,7-tetrahydrofuro[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole-4-carboxamide (I- 34):
Figure PCTCN2016087368-appb-000175
Figure PCTCN2016087368-appb-000175
按照制备化合物I-1的操作方法，以6-氟-2-(4,5,6,7-四氢呋喃并[2,3-c]吡啶-2- 基)-1H-苯并[d]咪唑-4-甲酸甲酯(4-18)(28mg，0.09mmol)为原料氨解得到灰白色粉末20mg，产率为70％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 6-fluoro-2-(4,5,6,7-tetrahydrofuro[2,3-c]pyridine-2- Methyl)-1H-benzo[d]imidazole-4-carboxylic acid methyl ester (4-18) (28 mg, 0.09 mmol) was obtained as a crude material to give 20 mg of an off-white powder (yield: 70%) :1).
1H NMR(300MHz,DMSO-d6)δ9.09(brs,1H),7.96(brs,1H),7.56(dd,J＝10.7,2.5Hz,1H),7.51(dd,J＝8.4,2.4Hz,1H),7.34(s,1H),4.16(s,2H),3.17(t,J＝5.0Hz,2H),2.68(t,J＝5.0Hz,2H)；13C NMR(151MHz,DMSO-d6)δ172.01,165.47,158.78(d,J＝237.0Hz,C-F),157.99,149.26,145.61,143.50,118.56,113.29,110.51(d,J＝27.0Hz,CH-CF),102.11,49.05,42.21,41.57；HRMS(ESI):m/z Calcd.For C15H13FN4O2[M+H]+:300.1023；Found:301.1093。 1 H NMR (300MHz, DMSO- d 6) δ9.09 (brs, 1H), 7.96 (brs, 1H), 7.56 (dd, J = 10.7,2.5Hz, 1H), 7.51 (dd, J = 8.4,2.4 Hz, 1H), 7.34 (s, 1H), 4.16 (s, 2H), 3.17 (t, J = 5.0 Hz, 2H), 2.68 (t, J = 5.0 Hz, 2H); 13 C NMR (151 MHz, DMSO) -d 6 ) δ 172.01, 165.47, 158.78 (d, J = 237.0 Hz, CF), 157.99, 149.26, 145.61, 143.50, 118.56, 113.29, 110.51 (d, J = 27.0 Hz, CH-CF), 102.11, 49.05, 42.21, 41.57; HRMS (ESI): m/z Calcd. For C 15 H 13 FN 4 O 2 [M+H] + : 300.1023; Found: 301.1093.
实施例35. 6-氟-2-(5-((甲氨基)甲基)噻吩-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-35)：Example 35. 6-Fluoro-2-(5-((methylamino)methyl)thiophen-2-yl)-1H-benzo[d]imidazole-4-carboxamide (I-35):
步骤1. ((5-溴噻吩-3-基)甲基)(甲基)氨基甲酸叔丁基酯：Step 1. ((5-Bromothiophen-3-yl)methyl)(methyl)carbamic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000176
Figure PCTCN2016087368-appb-000176
以5-溴噻吩-3-甲醛为原料，参照文献的方法经还原胺化得到的中间体经保护制得目标化合物(参考文献：Pyun,S.Y.；Lee,D.C.；Seung,Y.J.and Cho,B.R.Elimination Reactions of N-Alkyl-N-chlorothenylamines Promoted by MeONa-MeOH and Et2NH-MeCN.Effect of theβ-Aryl Group on the Imine-Forming Transition State.J.Org.Chem.2005,70,5327-5330.)。Using 5-bromothiophene-3-carbaldehyde as a raw material, the intermediate obtained by reductive amination according to the literature method is protected to obtain the target compound (Reference: Pyun, SY; Lee, DC; Seung, YJand Cho, BRElimination) Reactions of N-Alkyl-N-chlorothenylamines Promoted by MeONa-MeOH and Et2NH-MeCN. Effect of the β-Aryl Group on the Imine-Forming Transition State. J. Org. Chem. 2005, 70, 5327-5330.).
1H NMR(300MHz,Chloroform-d)δ6.87(d,J＝3.7Hz,1H),6.68(d,J＝3.7Hz,1H),4.44(s,2H),2.84(s,3H),1.49(s,9H)；MS(ESI)328[M+Na]+。 1 H NMR (300MHz, Chloroform- d) δ6.87 (d, J = 3.7Hz, 1H), 6.68 (d, J = 3.7Hz, 1H), 4.44 (s, 2H), 2.84 (s, 3H), 1.49 (s, 9H); MS (ESI) 328 [M+Na] + .
步骤2. ((5-甲酰基噻吩-2-基)甲基)(甲基)氨基甲酸叔丁酯：Step 2. ((5-Formylthiophen-2-yl)methyl)(methyl)carbamic acid tert-butyl ester:
Figure PCTCN2016087368-appb-000177
Figure PCTCN2016087368-appb-000177
按照制备化合物2-3的操作方法，以((5-溴噻吩-3-基)甲基)(甲基)氨基甲酸叔丁酯(12-14)(273mg，0.89mmol)为原料制备得到浅黄色液体208mg，产率为92％(石油醚：乙酸乙酯＝10：1)。Prepared according to the procedure for the preparation of compound 2-3, using ((5-bromothiophen-3-yl)methyl)(methyl)carbamic acid tert-butyl ester (12-14) (273 mg, 0.89 mmol) as a starting material The yellow liquid was 208 mg in a yield of 92% (petroleum ether: ethyl acetate = 10:1).
1H NMR(300MHz,Chloroform-d)δ9.86(s,1H),7.64(d,J＝3.8Hz,1H),7.02(d,J＝3.7Hz,1H),4.57(s,2H),2.89(s,3H),1.49(s,9H)；MS(ESI)278[M+Na]+。 1 H NMR (300 MHz, Chloroform-d) δ 9.86 (s, 1H), 7.64 (d, J = 3.8 Hz, 1H), 7.02 (d, J = 3.7 Hz, 1H), 4.57 (s, 2H), 2.89 (s, 3H), 1.49 (s, 9H); MS (ESI) 278 [M+Na] + .
步骤3. 2-(5-(((叔丁氧羰基)(甲基)氨基)甲基)噻吩-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯：Step 3. 2-(5-(((tert-Butoxycarbonyl)(methyl)amino)methyl)thiophen-2-yl)-6-fluoro-1H-benzo[d]imidazole-4-carboxylic acid methyl ester :
Figure PCTCN2016087368-appb-000178
Figure PCTCN2016087368-appb-000178
按照制备化合物3-7的操作方法，以((5-甲酰基噻吩-2-基)甲基)(甲基)氨基甲酸叔丁酯(2-15)(208mg，0.82mmol)和2,3-二氨基-5-氟苯甲酸甲酯(150mg，0.82mmol)为原料制备得到淡黄色固体333mg，产率为97％(石油醚：乙酸乙酯＝3：1)。According to the procedure for the preparation of compound 3-7, tert-butyl ((5-formylthiophen-2-yl)methyl)(methyl)carbamate (2-15) (208 mg, 0.82 mmol) and 2,3 Methyl 2-amino-5-fluorobenzoate (150 mg, 0.82 mmol) was obtained as a crude material (yield: 333 mg,yield: 97% (ethyl ether: ethyl acetate = 3:1).
1H NMR(300MHz,Chloroform-d)δ10.44(brs,1H),7.64(dd,J＝8.9,2.2Hz,1H),7.60(dd,J＝9.6,2.4Hz,1H),7.54(d,J＝3.7Hz,1H),7.00(d,J＝2.9Hz,1H),4.60(s,2H),4.02(s,3H),2.91(s,3H),1.50(s,9H)；MS(ESI)420[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 10.44 (brs, 1H), 7.64 (dd, J = 8.9, 2.2 Hz, 1H), 7.60 (dd, J = 9.6, 2.4 Hz, 1H), 7.54 (d) , J = 3.7 Hz, 1H), 7.00 (d, J = 2.9 Hz, 1H), 4.60 (s, 2H), 4.02 (s, 3H), 2.91 (s, 3H), 1.50 (s, 9H); (ESI) 420 [M+H] + .
步骤4. 6-氟-2-(5-((甲氨基)甲基)噻吩-2-基)-1H-苯并[d]咪唑-4-羧酸甲酯：Step 4. Methyl 6-fluoro-2-(5-((methylamino)methyl)thiophen-2-yl)-1H-benzo[d]imidazole-4-carboxylate:
Figure PCTCN2016087368-appb-000179
Figure PCTCN2016087368-appb-000179
按照制备化合物4-7的操作方法，以2-(5-(((叔丁氧羰基)(甲基)氨基)甲基)噻吩-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酸甲酯(3-19)(333mg，0.79mmol)为原料去保护基得到白色固体216mg，产率为86％(二氯甲烷：甲醇＝50：1)。According to the procedure for the preparation of compound 4-7, 2-(5-(((tert-butoxycarbonyl))(methyl)amino)methyl)thiophen-2-yl)-6-fluoro-1H-benzo[d] The imidazole-4-carboxylic acid methyl ester (3-19) (333 mg, 0.79 mmol) was obtained as a starting material to give a white solid 216 mg (yield: methylene chloride:methanol = 50:1).
1H NMR(300MHz,Chloroform-d)δ7.60(dd,J＝15.6,2.4Hz,1H),7.57(dd,J＝16.2,2.4Hz,1H),7.53(d,J＝3.4Hz,1H),6.97(d,J＝3.7Hz,1H),3.99(brs,5H),2.50(s,3H)；MS(ESI)320[M+H]+。 1 H NMR (300 MHz, Chloroform-d) δ 7.60 (dd, J = 15.6, 2.4 Hz, 1H), 7.57 (dd, J = 16.2, 2.4 Hz, 1H), 7.53 (d, J = 3.4 Hz, 1H) ), 6.97 (d, J = 3.7 Hz, 1H), 3.99 (brs, 5H), 2.50 (s, 3H); MS (ESI) 320 [M+H] + .
步骤5. 6-氟-2-(5-((甲氨基)甲基)噻吩-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-35)：Step 5. 6-Fluoro-2-(5-((methylamino)methyl)thiophen-2-yl)-1H-benzo[d]imidazole-4-carboxamide (I-35):
Figure PCTCN2016087368-appb-000180
Figure PCTCN2016087368-appb-000180
按照制备化合物I-1的操作方法，以6-氟-2-(4,5,6,7-四氢呋喃并[2,3-c]吡啶-2-基)-1H-苯并[d]咪唑-4-甲酸甲酯(4-19)(28mg，0.09mmol)为原料氨解得到灰白色粉末20mg，产率为70％(二氯甲烷：甲醇＝10：1)。According to the procedure for the preparation of compound I-1, 6-fluoro-2-(4,5,6,7-tetrahydrofuro[2,3-c]pyridin-2-yl)-1H-benzo[d]imidazole The methyl ester of 4-methyl formate (4-19) (28 mg, 0.09 mmol) was obtained as a crude material to give 20 mg of an off-white powder (yield: methylene chloride:methanol = 10:1).
1H NMR(300MHz,DMSO-d6)δ9.00(brs,1H),7.90(brs,1H),7.79(brs,1H),7.56-7.50(m,2H),7.08(d,J＝3.9Hz,1H),3.90(s,2H),2.33(s,3H)；13C NMR(125MHz,DMSO-d6)δ165.09,158.17(d,J＝236.6Hz,C-F),149.06,148.66,130.63,127.95,126.09,122.22,109.76(d,J＝26.2Hz,CH-CF),101.94(d,J＝27.1Hz,CH-CF),49.43,35.13；HRMS(ESI):m/z Calcd.For C14H14FN4OS[M+H]+:305.0872；Found:305.0863。 1 H NMR (300MHz, DMSO- d 6) δ9.00 (brs, 1H), 7.90 (brs, 1H), 7.79 (brs, 1H), 7.56-7.50 (m, 2H), 7.08 (d, J = 3.9 Hz, 1H), 3.90 (s, 2H), 2.33 (s, 3H); 13 C NMR (125MHz, DMSO-d 6 ) δ 165.09, 158.17 (d, J = 236.6 Hz, CF), 149.06, 148.66, 130.63, 127.95, 126.09, 122.22, 109.76 (d, J = 26.2 Hz, CH-CF), 101.94 (d, J = 27.1 Hz, CH-CF), 49.43, 35.13; HRMS (ESI): m/z Calcd. For C 14 H 14 FN 4 OS [M+H] + : 305.0872; Found: 305.0863.
实施例36. 2-(7-(2-氨基丙基-2-基)-4,5,6,7氢噻吩并[2,3-c]哌啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺(I-36)Example 36. 2-(7-(2-Aminopropyl-2-yl)-4,5,6,7-hydrothieno[2,3-c]piperidin-2-yl)-6-fluoro- 1H-benzo[d]imidazole-4-carboxamide (I-36)
步骤1. 2,3-二氨基-5-氟苯甲酰胺(36-1) Step 1. 2,3-Diamino-5-fluorobenzamide (36-1)
Figure PCTCN2016087368-appb-000181
Figure PCTCN2016087368-appb-000181
将2,3-二氨基-5-氟苯甲酸甲酯(920mg,0.5mmol)和NH3/MeOH溶液(8mol/L,50ml)加入到高压反应釜中，随后升温到100℃反应24h。冷却至室温，打开反应釜，将反应体系转移到单口瓶中浓缩除去甲醇得到粗产品。粗产品经柱层析(二氯甲烷：甲醇＝20：1)得430mg化合物36-1，产率为50％。The methyl 2,3-diamino-5-fluorobenzene (920mg, 0.5mmol) and NH 3 / MeOH solution (8mol / L, 50ml) was added to the autoclave, and then the reaction was heated to 100 deg.] C 24h. After cooling to room temperature, the reaction vessel was opened, and the reaction system was transferred to a single-mouth flask to concentrate to remove methanol to obtain a crude product. The crude product was subjected to column chromatography (dichloromethane:methanol = 20:1) to afford 430 g of Compound 36-1.
1H NMR(300MHz,CD3OD)δ6.72(dd,J＝2.1Hz,9.6Hz,1H),6.58(dd,J＝2.1Hz,9.6Hz,1H)；MS(ESI):m/z 170.1[M+H]+。 1 H NMR (300 MHz, CD 3 OD) δ 6.72 (dd, J = 2.1 Hz, 9.6 Hz, 1H), 6.58 (dd, J = 2.1 Hz, 9.6 Hz, 1H); MS (ESI): m/z 170.1 [M+H] + .
步骤2. 2-N-芴甲氧羰基氨基-2-甲基-N-(3-噻吩乙氨基)丙酰胺(36-2)Step 2. 2-N-Indolylmethoxycarbonylamino-2-methyl-N-(3-thienylethylamino)propanamide (36-2)
Figure PCTCN2016087368-appb-000182
Figure PCTCN2016087368-appb-000182
氮气保护下，将3-噻吩乙胺(440mg,3.5mmol)﹑2-N-芴甲氧羰基氨基-2-甲基丙酸(参见：Tetrahedron Letters,44(14),2807-2811；2003)(1.2g,3.8mmol)﹑HBTU(5.1g,13.4mmol)和碳酸钾(3.4g,24.4mmol)加入到无水N,N-二甲基甲酰胺(20ml)中，随后缓慢滴加三乙胺(1.35g,13.4mmol)于室温下搅拌直到主原料消失。将反应体系倒入到冰水中，用乙酸乙酯萃取的乙酸乙酯相，并干燥，浓缩得粗品。粗品快速过(二氯甲烷：甲醇＝20：1)得到900mg化合物36-2，收率60％。MS(ESI):m/z 434.9[M+H]+。3-Thienylethylamine (440 mg, 3.5 mmol), 2-N-fluorenylmethoxycarbonylamino-2-methylpropanoic acid under nitrogen (see: Tetrahedron Letters, 44 (14), 2807-2811; 2003) (1.2 g, 3.8 mmol), HBTU (5.1 g, 13.4 mmol) and potassium carbonate (3.4 g, 24.4 mmol) were added to anhydrous N,N-dimethylformamide (20 ml), followed by slow dropwise addition of triethyl The amine (1.35 g, 13.4 mmol) was stirred at room temperature until the main starting material disappeared. The reaction system was poured into ice water, and the ethyl acetate phase extracted with ethyl acetate. The crude product was quickly passed (dichloromethane:methanol = 20:1) to afford 900 mg of compound 36-2. MS (ESI): m / z 434.9 [M+H] + .
步骤3. 4-(2-(9-芴甲氧羰酰基氨基)丙-2-基)-6,7-二氢噻吩[3,2-c]吡啶-5(4H)-羧酸叔丁酯(36-3)Step 3. 4-(2-(9-Indolylmethoxycarbonylamino)propan-2-yl)-6,7-dihydrothiophene[3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl Ester (36-3)
Figure PCTCN2016087368-appb-000183
Figure PCTCN2016087368-appb-000183
按照制备化合物14-3的操作方法，以化合物2-N-芴甲氧羰基氨基-2-甲基-N-(3-噻吩乙氨基)丙酰胺(36-2)(8.3g,19.1mmol)为原料制得4.0g化合物36-3,直接用于下一步反应。The compound 2-N-fluorenylmethoxycarbonylamino-2-methyl-N-(3-thienylethylamino)propanamide (36-2) (8.3 g, 19.1 mmol) 4.0 g of compound 36-3 was obtained as a starting material and used directly for the next reaction.
步骤4. 2-溴-7-(2-(叔丁氧羰基氨基)丙基-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)-羧酸叔丁酯(36-4)Step 4. 2-Bromo-7-(2-(tert-butoxycarbonylamino)propyl-2-yl)-4,5-dihydrothieno[2,3-c]pyridine-6(7H)-carboxylate Tert-butyl acid ester (36-4)
Figure PCTCN2016087368-appb-000184
Figure PCTCN2016087368-appb-000184
a)将4-(2-(9-芴甲氧羰酰基氨基)丙-2-基)-6,7-二氢噻吩[3,2-c]吡啶-5(4H)-羧酸叔 丁酯(36-3)(1.0g,2.0mmol)和NBS(390.0mg,2.2mmol)加入到二氯甲烷中于室温下反应直到主原料消失。快速过柱(PE:EA＝10:1)得到900mg溴代物。b)将上述a)步溴代物(300mg,0.5mmol)溶于二氯甲烷中,随后加入三乙胺(0.5当量)于回流下反应直到完全脱除9-芴甲氧羰酰基。快速过柱(DCM:MeOH＝100～20:1)得到156mg中间体，MS(ESI)374.8/376.9(1:1)[M+H]+。c)将b)步产物(156mg,0.42mmol)加入到四氢呋喃(10ml)中，随后加入二碳酸二叔丁酯(109mg,0.5mmol)于回流状态下反应直到主原料消失。将反应体系浓缩，快速过柱(石油醚：乙酸乙酯＝10：1)得到160mg化合物36-4。MS(ESI):m/z497.2,499.1[M+Na]+。a) 4-(2-(9-fluorenylmethoxycarbonylamino)propan-2-yl)-6,7-dihydrothiophene [3,2-c]pyridine-5(4H)-carboxylic acid tert-butyl Ester (36-3) (1.0 g, 2.0 mmol) and NBS (390.0 mg, 2.2 mmol) were added to dichloromethane and reacted at room temperature until the main material disappeared. Fast column (PE: EA = 10:1) gave 900 mg of bromo. b) The bromo compound of the above step a) (300 mg, 0.5 mmol) was dissolved in dichloromethane, then triethylamine (0.5 eq.) was added and refluxed until reflux. The column was quickly passed (DCM: MeOH = 100 to 20:1) to afford 156 mg of Intermediate, MS (ESI) 374.8/376.9 (1:1) [M+H] + . c) The product from step b) (156 mg, 0.42 mmol) was added to tetrahydrofuran (10 ml), and then di-tert-butyl dicarbonate (109 mg, 0.5 mmol) was added to react under reflux until the main material disappeared. The reaction system was concentrated and rapidly passed through a column (peel ether: ethyl acetate = 10:1) to afford 160 mg of Compound 36-4. MS (ESI): m / z497.2,499.1 [M + Na] +.
步骤5. 7-(2-(叔丁氧羰基氨基)丙基-2-基)-2-甲酰基-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)-羧酸叔丁酯(36-5)Step 5. 7-(2-(tert-Butoxycarbonylamino)propyl-2-yl)-2-formyl-4,5-dihydrothieno[2,3-c]pyridine-6(7H)- Tert-butyl carboxylate (36-5)
Figure PCTCN2016087368-appb-000185
Figure PCTCN2016087368-appb-000185
按照制备化合物2-3的操作方法,以2-溴-7-(2-(叔丁氧羰基氨基)丙基-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)-羧酸叔丁酯(36-4)(100.0mg,0.21mmol)为原料制得40mg化合物36-5，收率44％。MS(ESI):m/z 446.9[M+Na]+。According to the procedure for the preparation of compound 2-3, 2-bromo-7-(2-(tert-butoxycarbonylamino)propyl-2-yl)-4,5-dihydrothieno[2,3-c] Pyridine-6(7H)-carboxylic acid tert-butyl ester (36-4) (100.0 mg, 0.21 mmol) was used as a starting material to afford 40 mg of Compound 36-5. MS (ESI): m/z 446.9 [M+Na] + .
步骤6. 7-(2-(叔丁氧羰基氨基)丙基-2-基)-2-(4-氨甲酰基-6-氟-1H-苯并[d]咪唑-2-基)-4,5-二氢噻吩[2,3-c]吡啶-6(7H)-羧酸叔丁酯(36-6)Step 6. 7-(2-(tert-Butoxycarbonylamino)propyl-2-yl)-2-(4-carbamoyl-6-fluoro-1H-benzo[d]imidazol-2-yl)- 4,5-dihydrothiophene [2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (36-6)
Figure PCTCN2016087368-appb-000186
Figure PCTCN2016087368-appb-000186
将2,3-二氨基-5-氟苯甲酰胺(36-1)(68mg,0.36mmol)、化合物36-5(152.0mg,0.36mmol)和亚硫酸氢钠(684mg,3.6mmol)加入到乙醇(20.0ml)中于60℃下反应直到原料反应完全。将反应体系浓缩除去乙醇。随后加入水和二氯甲烷进行搅拌溶解，静置、萃取、分液的二氯甲烷相。二氯甲烷相干燥、浓缩得粗品。粗产品经柱层析(二氯甲烷：甲醇＝20：1)得168mg化合物36-6，收率81.5％。1H NMR(300MHz,CD3OD)δ7.65(dd,J＝2.7,10.8Hz),7.50(s,1H),7.39(dd,J＝2.7,8.4Hz),6.25(s,1H),4.46～4.35(m,1H),3.17(brs,1H),2.70～2.65(m,2H),1.56(s,9H),1.54(s,9H),1.29(s,6H)；MS(ESI):m/z574.36[M+H]+。2,3-Diamino-5-fluorobenzamide (36-1) (68 mg, 0.36 mmol), compound 36-5 (152.0 mg, 0.36 mmol) and sodium hydrogen sulfite (684 mg, 3.6 mmol) were added Ethanol (20.0 ml) was reacted at 60 ° C until the starting material was completely reacted. The reaction system was concentrated to remove ethanol. Subsequently, water and dichloromethane were added to stir and dissolve, and the dichloromethane phase was allowed to stand, extracted, and separated. The methylene chloride phase was dried and concentrated to give a crude material. The crude product was subjected to column chromatography (dichloromethane:methanol = 20:1) to yield 168 mg of Compound 36-6. 1 H NMR (300 MHz, CD 3 OD) δ 7.65 (dd, J = 2.7, 10.8 Hz), 7.50 (s, 1H), 7.39 (dd, J = 2.7, 8.4 Hz), 6.25 (s, 1H), 4.46 to 4.35 (m, 1H), 3.17 (brs, 1H), 2.70 to 2.65 (m, 2H), 1.56 (s, 9H), 1.54 (s, 9H), 1.29 (s, 6H); MS (ESI) :m/z574.36[M+H] + .
步骤7. 2-(7-(2-氨基丙基-2-基)-4,5,6,7氢噻吩并[2,3-c]哌啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺(I-36) Step 7. 2-(7-(2-Aminopropyl-2-yl)-4,5,6,7-hydrothieno[2,3-c]piperidin-2-yl)-6-fluoro-1H -benzo[d]imidazole-4-carboxamide (I-36)
Figure PCTCN2016087368-appb-000187
Figure PCTCN2016087368-appb-000187
将7-(2-(叔丁氧羰基氨基)丙基-2-基)-2-(4-氨甲酰基-6-氟-1H-苯并[d]咪唑-2-基)-4,5-二氢噻吩[2,3-c]吡啶-6(7H)-羧酸叔丁酯(36-6)(76.0mg,0.13mmol)和三氟乙酸(0.5mL)加入到二氯甲烷(20.0mL)中于室温下搅拌直到原料反应完全。将反应体系浓缩的粗产品，粗产品经柱层析(二氯甲烷：甲醇＝20：1)得24mg化合物I-36,收率48％。1H NMR(300MHz,d6-DMSO)δ7.95(brs,1H),7.62(s,1H),7.58(s,1H),5.1(s,1H),3.60(m,1H),3.26～3.12(m,2H),2.98(m,1H),1.70(s,3H),1.65(s,3H)；MS(ESI):m/z 374.36[M+H]+ 7-(2-(tert-Butoxycarbonylamino)propyl-2-yl)-2-(4-carbamoyl-6-fluoro-1H-benzo[d]imidazol-2-yl)-4, 5-Dihydrothiophene [2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (36-6) (76.0 mg, 0.13 mmol) and trifluoroacetic acid (0.5 mL) were added to dichloromethane ( Stir at room temperature in 20.0 mL) until the starting material was completely reacted. The crude product which was concentrated in the reaction system was purified by column chromatography (dichloromethane:methanol = 20:1) to afford 24 mg of Compound I-36. 1 H NMR (300MHz, d 6 -DMSO) δ7.95 (brs, 1H), 7.62 (s, 1H), 7.58 (s, 1H), 5.1 (s, 1H), 3.60 (m, 1H), 3.26 ~ 3.12 (m, 2H), 2.98 (m, 1H), 1.70 (s, 3H), 1.65 (s, 3H); MS (ESI): m/z 374.36 [M+H] +
实施例37. 6-氟-2-(7-(2-羟基丙基-2-基))-4,5,6,7-四氢噻吩并[2,3-c]哌啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-37)Example 37. 6-Fluoro-2-(7-(2-hydroxypropyl-2-yl))-4,5,6,7-tetrahydrothieno[2,3-c]piperidin-2- -1H-benzo[d]imidazole-4-carboxamide (I-37)
步骤1. 2-甲基-1-氧代-1-(2-(噻吩-3基)乙胺基)丙-2基-4-甲氧基苯甲酸(37-1)Step 1. 2-Methyl-1-oxo-1-(2-(thiophen-3-yl)ethylamino)propan-2-yl-4-methoxybenzoic acid (37-1)
Figure PCTCN2016087368-appb-000188
Figure PCTCN2016087368-appb-000188
氮气保护下，将3-噻吩乙胺(1.55g,12.2mmol)﹑2-((4-甲氧基苯甲酰氧基)2-甲基丙酸(2.9g,12.2mmol)﹑HBTU(5.1g,13.4mmol)和碳酸钾(3.4g,24.4mmol)加入到无水N,N-二甲基甲酰胺(20ml)中，随后缓慢滴加三乙胺(1.35g,13.4mmol)于室温下搅拌直到主原料消失。将反应体系倒入到冰水中，用乙酸乙酯萃取的乙酸乙酯相，并干燥，浓缩得粗品。粗品快速过(二氯甲烷：甲醇＝20：1)得到2.4g化合物37-1，收率57％。MS(ESI):m/z 348.2[M+H]+ 3-Thienylethylamine (1.55 g, 12.2 mmol), 2-((4-methoxybenzoyloxy) 2-methylpropionic acid (2.9 g, 12.2 mmol), HBTU (5.1) g, 13.4 mmol) and potassium carbonate (3.4 g, 24.4 mmol) were added to anhydrous N,N-dimethylformamide (20 ml), then triethylamine (1.35 g, 13.4 mmol) was slowly added dropwise at room temperature Stirring until the main starting material disappeared. The reaction system was poured into ice water, and the ethyl acetate phase was extracted with ethyl acetate, and dried and concentrated to give a crude product. The crude product was quickly passed (dichloromethane:methanol = 20:1) to give 2.4 g. Compound 37-1, yield 57%. MS (ESI): m/z 348.2 [M+H] +
步骤2. 7-(2-(4-甲氧苯甲酰氧基)-丙-2-基)-4,5-二氢噻吩并(2,3-c)吡啶-6(7H)-羧酸叔丁酯(37-2)Step 2. 7-(2-(4-Methoxybenzoyloxy)-propan-2-yl)-4,5-dihydrothieno(2,3-c)pyridine-6(7H)-carboxylate Tert-butyl acid (37-2)
Figure PCTCN2016087368-appb-000189
Figure PCTCN2016087368-appb-000189
按照制备化合物14-3的操作方法，以2-甲基-1-氧代-1-(2-(噻吩-3基)乙胺基)丙-2基-4-甲氧基苯甲酸(37-1)(1.3g,3.7mmol)为原料得到1.1g化合物37-2，收率62.5％。1H NMR(300MHz,CDCl3)δ7.38(d,J＝81.Hz,2H),7.21(d,J＝5.1Hz,1H),6.93(d,J＝8.1Hz,2H),6.79(d,J＝5.1Hz,1H),6.20(s,1H),4.00(d,J＝12.3Hz,1H),3.84(s,3H),3.62～3.77(m,1H),2.61(brs,2H),1.82(s,3H),1.56(s,3H),1.45(s,9H)；MS(ESI):m/z 454[M+Na]+。 According to the procedure for the preparation of compound 14-3, 2-methyl-1-oxo-1-(2-(thiophen-3-yl)ethylamino)propan-2-yl-4-methoxybenzoic acid (37) -1) (1.3 g, 3.7 mmol) was used as a starting material to obtain 1.1 g of Compound 37-2, yield 62.5%. 1 H NMR (300MHz, CDCl 3 ) δ7.38 (d, J = 81.Hz, 2H), 7.21 (d, J = 5.1Hz, 1H), 6.93 (d, J = 8.1Hz, 2H), 6.79 ( d, J = 5.1 Hz, 1H), 6.20 (s, 1H), 4.00 (d, J = 12.3 Hz, 1H), 3.84 (s, 3H), 3.62 to 3.77 (m, 1H), 2.61 (brs, 2H) ), 1.82 (s, 3H), 1.56 (s, 3H), 1.45 (s, 9H); MS (ESI): m/z 454 [M+Na] + .
步骤3. 2-溴-7-(2-(4-甲氧苯甲酰氧基)-丙-2-基)-4,5-二氢噻吩并(2,3-c)吡啶-6(7H)-羧酸叔丁酯(37-3)Step 3. 2-Bromo-7-(2-(4-methoxybenzoyloxy)-propan-2-yl)-4,5-dihydrothieno(2,3-c)pyridine-6 ( 7H)-tert-butyl carboxylic acid (37-3)
Figure PCTCN2016087368-appb-000190
Figure PCTCN2016087368-appb-000190
按照制备化合物12-3的制备方法，以7-(2-(4-甲氧苯甲酰氧基)-丙-2-基)-4,5-二氢噻吩并(2,3-c)吡啶-6(7H)-羧酸叔丁酯(37-2)(1.2g,2.78mmol)为原料得到1.35g化合物37-3，收率95％。1H NMR(400MHz,CDCl3)δ7.37(d,J＝10.4Hz,2H),6.93(d,J＝10.4Hz,2H),6.75(s,1H),6.07(s,1H),4.00(d,1H),3.60～3.67(m,1H),2.52(brs,2H),1.76(s,3H),1.57(s,1H),1.47(s,9H)；MS(ESI):m/z 532.1,534.1[M+Na]+。According to the preparation method of the compound 12-3, 7-(2-(4-methoxybenzoyloxy)-propan-2-yl)-4,5-dihydrothieno(2,3-c) Pyridine-6(7H)-carboxylic acid tert-butyl ester (37-2) (1.2 g, 2.78 mmol) was used as a starting material to give 1.35 g of Compound 37-3. 1 H NMR (400MHz, CDCl 3 ) δ7.37 (d, J = 10.4Hz, 2H), 6.93 (d, J = 10.4Hz, 2H), 6.75 (s, 1H), 6.07 (s, 1H), 4.00 (d, 1H), 3.60 to 3.67 (m, 1H), 2.52 (brs, 2H), 1.76 (s, 3H), 1.57 (s, 1H), 1.47 (s, 9H); MS (ESI): m/ z 532.1, 534.1 [M+Na] + .
步骤4. 2-甲酰基-7-(2-羟基丙基-2-基)-4,5-二氢噻吩并[2,3,-c]吡啶-6(7H)甲酸叔丁酯(37-4)Step 4. 2-Formyl-7-(2-hydroxypropyl-2-yl)-4,5-dihydrothieno[2,3,-c]pyridine-6(7H)carboxylic acid tert-butyl ester (37 -4)
Figure PCTCN2016087368-appb-000191
Figure PCTCN2016087368-appb-000191
-78℃，在氮气保护下，往2-溴-7-(2-(4-甲氧苯甲酰氧基)-丙-2-基)-4,5-二氢噻吩并(2,3-c)吡啶-6(7H)-羧酸叔丁酯(37-3)(255.0mg，0.5mmol)的无水四氢呋喃(20mL)溶液中缓慢滴加3.8mL正丁基锂的四氢呋喃溶液(1.6M)，继续搅拌1小时。往反应液中加入无水N,N-二甲基甲酰胺(0.3mL)，搅拌1小时后，加入1mL水淬灭。反应液用乙酸乙酯萃取(50mL×3)，有机层依次用水洗，饱和食盐水洗，保留有机层，用无水硫酸钠干燥，过滤减压浓缩，粗产品经柱层析(石油醚：乙酸乙酯＝10：1)得43mg化合物37-4，产率为18％。1H NMR(400MHz,CDCl3)δ9.85(s,1H),7.50(s,1H),5.28(s,0.5H),5.12(s,0.5H),4.50～4.23(m,0.5H),3.91～3.75(m,1H),3.36～3.21(m,0.5H),2.68(m,2H),1.48(s,9H),1.46(s,3H),1.25(s,3H)。MS(ESI):m/z 347.9[M+Na]+。2-bromo-7-(2-(4-methoxybenzoyloxy)-propan-2-yl)-4,5-dihydrothieno(2,3) at -78 ° C under N2 -c) pyridine hydrochloride-6(7H)-carboxylic acid tert-butyl ester (37-3) (255.0 mg, 0.5 mmol) in anhydrous tetrahydrofuran (20 mL) was slowly added dropwise 3.8 mL of n-butyllithium in tetrahydrofuran (1.6) M), continue to stir for 1 hour. Anhydrous N,N-dimethylformamide (0.3 mL) was added to the reaction mixture, and stirred for 1 hour, then quenched with 1 mL of water. The reaction mixture was extracted with ethyl acetate (50 mL×3). EtOAc. Ethyl ester = 10:1) gave 43 mg of compound 37-4 in 18% yield. 1 H NMR (400 MHz, CDCl 3 ) δ 9.85 (s, 1H), 7.50 (s, 1H), 5.28 (s, 0.5H), 5.12 (s, 0.5H), 4.50 to 4.23 (m, 0.5H) , 3.91 to 3.75 (m, 1H), 3.36 to 3.21 (m, 0.5H), 2.68 (m, 2H), 1.48 (s, 9H), 1.46 (s, 3H), 1.25 (s, 3H). MS (ESI): m / z 347.9 [M + Na] +.
步骤5. 2-(6-氟-4-(甲氧羰基)-1H-苯并[d]咪唑-2-基)-7-(2-羟基丙基-2-基)-4,5-二氢噻吩并[2,3,-c]吡啶-6(7H)甲酸叔丁酯(37-5)Step 5. 2-(6-Fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-7-(2-hydroxypropyl-2-yl)-4,5- Dihydrothieno[2,3,-c]pyridine-6(7H)carboxylic acid tert-butyl ester (37-5)
Figure PCTCN2016087368-appb-000192
Figure PCTCN2016087368-appb-000192
按照制备化合物3-7的操作方法，以2-甲酰基-7-(2-羟基丙基-2-基)-4,5-二氢噻吩并[2,3,-c]吡啶-6(7H)甲酸叔丁酯(37-4)(130.0mg,0.4mmol)和2,3-二氨基-5-氟苯甲酸甲酯(73.5mg，0.4mmol)为原料得到131mg固体化合物37-5，收率66.9％。1H NMR(300MHz, d4-CD3OD)δ7.74(s,1H),7.62(dd,J＝1.8,7.2Hz),7.55(d,J＝6.3Hz,1H),5.25(s,0.5H),5.16(s,0.5H),4.42～4.29(m,1H),4.09(s,3H),3.43～3.38(m,0.5H),3.28～3.40(m,0.5H),2.72(m,2H),1.51(s,9H),1.45(d,J＝5.4Hz,3H),1.19(s,3H)；MS(ESI):m/z 489.9[M+H]+。According to the procedure for the preparation of compound 3-7, 2-formyl-7-(2-hydroxypropyl-2-yl)-4,5-dihydrothieno[2,3,-c]pyridine-6 ( 7H) tert-butyl formate (37-4) (130.0 mg, 0.4 mmol) and 2,3-diamino-5-fluorobenzoic acid methyl ester (73.5 mg, 0.4 mmol) The yield was 66.9%. 1 H NMR (300MHz, d 4 -CD 3 OD) δ7.74 (s, 1H), 7.62 (dd, J = 1.8,7.2Hz), 7.55 (d, J = 6.3Hz, 1H), 5.25 (s, 0.5H), 5.16 (s, 0.5H), 4.42 to 4.29 (m, 1H), 4.09 (s, 3H), 3.43 to 3.38 (m, 0.5H), 3.28 to 3.40 (m, 0.5H), 2.72 ( m, 2H), 1.51 (s, 9H), 1.45 (d, J = 5.4 Hz, 3H), 1.19 (s, 3H); MS (ESI): m/z 489.9 [M+H] + .
步骤6. 6-氟-2-(7-(2-羟基丙基-2-基))-4,5,6,7-四氢噻吩并[2,3-c]哌啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-37)Step 6. 6-Fluoro-2-(7-(2-hydroxypropyl-2-yl))-4,5,6,7-tetrahydrothieno[2,3-c]piperidin-2-yl )-1H-benzo[d]imidazole-4-carboxamide (I-37)
Figure PCTCN2016087368-appb-000193
Figure PCTCN2016087368-appb-000193
将2-(6-氟-4-(甲氧羰基)-1H-苯并[d]咪唑-2-基)-7(2-羟基丙-2-基)-4,5-二氢噻吩并[2,3,-c]吡啶-6(7H)甲酸叔丁酯(37-5)(125.0mg,0.25mmol)和氨的甲醇溶液(10mol/L,30mL)加入到高压反应釜中于100℃下反应直到原料完全反应。冷却，浓缩得到粗品酰胺。将粗品酰胺溶于二氯甲烷中，随后加入三氟乙酸(2mL)于室温下反应直到原料完全消失，浓缩，粗品快速过(二氯甲烷：甲醇＝20：1)得到25mg化合物I-37，收率26％。1H NMR(300MHz,d4-CD3OD)δ7.64(dd,J＝10.4Hz,J＝1.8Hz,1H),7.51(s,1H),7.39(dd,J＝8.8Hz,J＝1.8Hz,1H),4.18(s,1H),3.45(m,1H),3.11～3.04(m,1H),2.90～2.77(m,2H),1.41(s,3H),1.26(s,3H)；MS(ESI):m/z 374.9[M+H]+。2-(6-Fluoro-4-(methoxycarbonyl)-1H-benzo[d]imidazol-2-yl)-7(2-hydroxypropan-2-yl)-4,5-dihydrothiophene [2,3,-c]pyridine-6(7H)carboxylic acid tert-butyl ester (37-5) (125.0 mg, 0.25 mmol) and ammonia in methanol (10 mol/L, 30 mL) were added to a high pressure reaction kettle at 100 The reaction was carried out at ° C until the starting material was completely reacted. Cool and concentrate to give the crude amide. The crude amide was dissolved in dichloromethane, then trifluoroacetic acid (2 mL) was added at room temperature until the material was completely evaporated, concentrated, and the crude product was purified (dichloromethane:methanol = 20:1) The yield was 26%. 1 H NMR (300MHz, d 4 -CD 3 OD) δ7.64 (dd, J = 10.4Hz, J = 1.8Hz, 1H), 7.51 (s, 1H), 7.39 (dd, J = 8.8Hz, J = 1.8 Hz, 1H), 4.18 (s, 1H), 3.45 (m, 1H), 3.11 to 3.04 (m, 1H), 2.90 to 2.77 (m, 2H), 1.41 (s, 3H), 1.26 (s, 3H) MS (ESI): m/z 374.9 [M+H] + .
实施例38. 2-(2-(4-氨基甲酰基-6-氟-1H-苯并[d]咪唑-2-基)-4,5,6,7-四氢噻吩并[2,3-c]哌啶-7-基)-2-异丙酸甲酯(I-38)Example 38. 2-(2-(4-Aminoformyl-6-fluoro-1H-benzo[d]imidazol-2-yl)-4,5,6,7-tetrahydrothieno[2,3 -c] piperidine-7-yl)-2-isopropylate (I-38)
步骤1. 2,2-二甲基-3-氧代-3-(2-(噻吩-3-基)二乙胺基)丙酸甲酯(38-1)Step 1. 2,2-Dimethyl-3-oxo-3-(2-(thien-3-yl)diethylamino)propanoic acid methyl ester (38-1)
Figure PCTCN2016087368-appb-000194
Figure PCTCN2016087368-appb-000194
按照制备化合物37-1的操作方法，以3-噻吩乙胺(870.0mg,6.84mmol)和2,2-二甲基丙二酸单甲酯(1.0g,6.84mmol)为原料制得1.4g油状化合物38-1，收率80％。1H NMR(300MHz,CDCl3)δ7.27(d,J＝5.1Hz,1H),6.98(s,1H),6.93(d,J＝5.1Hz,1H),6.34(brs,1H),3.67(s,3H),3.49(q,J＝6.6Hz,2H),2.84(t,J＝6.6Hz,2H),1.40(s,6H)；MS(ESI):m/z256.1[M+H]+。1.4 g of 3-thienoethylamine (870.0 mg, 6.84 mmol) and 2,2-dimethylmalonic acid monomethyl ester (1.0 g, 6.84 mmol) were prepared according to the procedure for the preparation of compound 37-1. Oily compound 38-1, yield 80%. 1 H NMR (300MHz, CDCl 3 ) δ7.27 (d, J = 5.1Hz, 1H), 6.98 (s, 1H), 6.93 (d, J = 5.1Hz, 1H), 6.34 (brs, 1H), 3.67 (s, 3H), 3.49 (q, J = 6.6 Hz, 2H), 2.84 (t, J = 6.6 Hz, 2H), 1.40 (s, 6H); MS (ESI): m/z 256.1 [M+ H] + .
步骤2. 7-(1-甲氧基-2-甲基-1-氧代丙基-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)羧酸叔丁酯(38-2)Step 2. 7-(1-Methoxy-2-methyl-1-oxopropyl-2-yl)-4,5-dihydrothieno[2,3-c]pyridine-6(7H) Tert-butyl carboxylate (38-2)
Figure PCTCN2016087368-appb-000195
Figure PCTCN2016087368-appb-000195
按照制备化合物14-3的操作方法，以2,2-二甲基-3-氧代-3-(2-(噻吩-3-基)二乙胺基)丙酸甲酯(38-1)(1.27g,5.0mmol)为原料制得170mg化合物38-2，收率10％。1H NMR(300MHz,CDCl3)δ7.13(d,J＝4.8Hz,1H),6.78(d,J＝4.8Hz,1H),5.82(s,1H),4.36(m,1H),3.75(s,3H),3.15(m,1H),2.61(m,2H),1.48(s,9H),1.35(s,3H),1.19(s,3H)；MS(ESI):m/z 240.1[M+H]+.According to the procedure for the preparation of compound 14-3, methyl 2,2-dimethyl-3-oxo-3-(2-(thien-3-yl)diethylamino)propanoate (38-1) (1.27 g, 5.0 mmol) was used as a starting material to obtain 170 mg of compound 38-2 in a yield of 10%. 1 H NMR (300MHz, CDCl 3 ) δ7.13 (d, J = 4.8Hz, 1H), 6.78 (d, J = 4.8Hz, 1H), 5.82 (s, 1H), 4.36 (m, 1H), 3.75 (s, 3H), 3.15 (m, 1H), 2.61 (m, 2H), 1.48 (s, 9H), 1.35 (s, 3H), 1.19 (s, 3H); MS (ESI): m/z 240.1 [M+H] + .
步骤3. 2-溴-7-(1-甲氧基-2-甲基-1-氧代丙基-2基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)羧酸叔丁酯(38-3)Step 3. 2-Bromo-7-(1-methoxy-2-methyl-1-oxopropyl-2yl)-4,5-dihydrothieno[2,3-c]pyridine-6 (7H) tert-butyl carboxylate (38-3)
Figure PCTCN2016087368-appb-000196
Figure PCTCN2016087368-appb-000196
按照制备化合物12-3的制备方法,以7-(1-甲氧基-2-甲基-1-氧代丙基-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)羧酸叔丁酯(38-2)(170mg,0.5mmol)为原料制得132mg化合物38-3，收率63％。MS(ESI):m/z 440.1；442.1[M+Na]+。According to the preparation method of the compound 12-3, 7-(1-methoxy-2-methyl-1-oxopropyl-2-yl)-4,5-dihydrothieno[2,3- c] Pyridyl-6(7H)carboxylic acid tert-butyl ester (38-2) (170 mg, 0.5 mmol) was obtained as a material. MS (ESI): m / z 440.1; 442.1 [M + Na] +.
步骤4. 2-甲酰基-7-(1-甲氧基-2-甲基-1-氧代丙基-2基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)羧酸叔丁酯(38-4)Step 4. 2-Formyl-7-(1-methoxy-2-methyl-1-oxopropyl-2yl)-4,5-dihydrothieno[2,3-c]pyridine- 6(7H)carboxylic acid tert-butyl ester (38-4)
Figure PCTCN2016087368-appb-000197
Figure PCTCN2016087368-appb-000197
按照制备化合物2-3的操作方法,以2-溴-7-(1-甲氧基-2-甲基-1-氧代丙基-2基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)羧酸叔丁酯(38-3)(132.0mg,0.32mmol)为原料制得55mg化合物38-4，收率47％。1H NMR(300MHz,CDCl3)δ9.80(s,1H,CHO),7.45(s,1H),5.87(s,1H),4.50～4.26(m,1H),3.75(s,3H),3.16(m,1H),2.65(m,2H),1.46(s,9H),1.35(s,3H),1.17(s,3H)；MS(ESI):m/z 380.8[M+Na]+.According to the procedure for the preparation of compound 2-3, 2-bromo-7-(1-methoxy-2-methyl-1-oxopropyl-2yl)-4,5-dihydrothieno[2] , 3-c]pyridine-6(7H)carboxylic acid tert-butyl ester (38-3) (132.0 mg, 0.32 mmol) was obtained. 1 H NMR (300MHz, CDCl 3 ) δ9.80 (s, 1H, CHO), 7.45 (s, 1H), 5.87 (s, 1H), 4.50 ~ 4.26 (m, 1H), 3.75 (s, 3H), 3.16(m,1H), 2.65(m,2H), 1.46(s,9H), 1.35(s,3H), 1.17(s,3H);MS(ESI):m/z 380.8[M+Na] + .
步骤5. 2-(4-氨甲酰基-6-氟-1H-苯并[d]咪唑-2-基)-7-(1-甲氧基-2-甲基-1-氧代丙基-2基)-4,5-二氢噻吩[2,3,c]吡啶-6(7H)羧酸叔丁酯(38-5)Step 5. 2-(4-carbamoyl-6-fluoro-1H-benzo[d]imidazol-2-yl)-7-(1-methoxy-2-methyl-1-oxopropyl -2yl)-4,5-dihydrothiophene [2,3,c]pyridine-6(7H)carboxylic acid tert-butyl ester (38-5)
Figure PCTCN2016087368-appb-000198
Figure PCTCN2016087368-appb-000198
将2,3-二氨基-5-氟苯甲酰胺(36-1)(30.4mg,0.18mmol)、化合物(38-4)(55.0mg,0.15mmol)和亚硫酸氢钠(156.0mg,1.5mmol)加入到乙醇(20.0ml)中于60℃下反应直到原料反应完全。将反应体系浓缩除去乙醇。随后加入水和二氯甲烷进行搅拌溶解，静置、萃取、分液的二氯甲烷相。二氯甲烷相干燥、浓缩得粗品。粗产品经柱层析(二氯 甲烷：甲醇＝20：1)得68mg化合物38-5，收率87.7％。1H NMR(300MHz,CDCl3)δ11.94(brs,1H,NH2),9.72(brs,1H,NH2),7.74(brs,1H),7.43(s,1H),7.26(brs,1H),6.29(brs,1H,NH),5.81(brs,1H),4.35(s,1H),3.68(s,3H),3.18(m,1H),2.56(m,2H),1.46(s,9H),1.35(s,3H),1.20(s,3H)；MS(ESI):m/z 316.8[M+H]+。2,3-Diamino-5-fluorobenzamide (36-1) (30.4 mg, 0.18 mmol), compound (38-4) (55.0 mg, 0.15 mmol) and sodium hydrogen sulfite (156.0 mg, 1.5) Methyl) was added to ethanol (20.0 ml) at 60 ° C until the starting material was completely reacted. The reaction system was concentrated to remove ethanol. Subsequently, water and dichloromethane were added to stir and dissolve, and the dichloromethane phase was allowed to stand, extracted, and separated. The methylene chloride phase was dried and concentrated to give a crude material. The crude product was subjected to column chromatography (dichloromethane:methanol = 20:1) to yield 68 mg of Compound 38-5. 1 H NMR (300MHz, CDCl 3 ) δ11.94 (brs, 1H, NH 2), 9.72 (brs, 1H, NH 2), 7.74 (brs, 1H), 7.43 (s, 1H), 7.26 (brs, 1H ), 6.29 (brs, 1H, NH), 5.81 (brs, 1H), 4.35 (s, 1H), 3.68 (s, 3H), 3.18 (m, 1H), 2.56 (m, 2H), 1.46 (s, 9H), 1.35 (s, 3H), 1.20 (s, 3H); MS (ESI): m/z 316.8 [M+H] + .
步骤6. 2-(2-(4-氨基甲酰基-6-氟-1H-苯并[d]咪唑-2-基)-4,5,6,7-四氢噻吩并[2,3-c]哌啶-7-基)-2-异丙酸甲酯(I-38)Step 6. 2-(2-(4-Aminoformyl-6-fluoro-1H-benzo[d]imidazol-2-yl)-4,5,6,7-tetrahydrothieno[2,3- c] methyl piperidine-7-yl)-2-isopropylate (I-38)
Figure PCTCN2016087368-appb-000199
Figure PCTCN2016087368-appb-000199
将2-(4-氨甲酰基-6-氟-1H-苯并[d]咪唑-2-基)-7-(1-甲氧基-2-甲基-1-氧代丙基-2基)-4,5-二氢噻吩[2,3,c]吡啶-6(7H)羧酸叔丁酯(38-5)(68.0mg,0.13mmol)和三氟乙酸(0.5mL)加入到二氯甲烷(20.0mL)中于室温下搅拌直到原料反应完全，将反应体系浓缩的粗产品。粗产品经柱层析(二氯甲烷：甲醇＝20：1)得20mg化合物I-38,收率36％。1H NMR(300MHz,CD3OD)δ7.64(d,1H),7.47(s,1H),7.34(d,1H),4.55(s,1H),3.80(s,3H),3.35(s,1H),2.97～2.90(m,1H),2.79～2.67(m,2H),1.26(s,6H)；MS(ESI):m/z 417.2[M+H]+。2-(4-carbamoyl-6-fluoro-1H-benzo[d]imidazol-2-yl)-7-(1-methoxy-2-methyl-1-oxopropyl-2 Tert-butyl-4,5-dihydrothiophene [2,3,c]pyridine-6(7H)carboxylate (38-5) (68.0 mg, 0.13 mmol) and trifluoroacetic acid (0.5 mL) The crude product was concentrated in dichloromethane (20.0 mL) at room temperature until the starting material was completely reacted. The crude product was subjected to column chromatography (dichloromethane:methanol = 20:1) to yield 20 mg of Compound I-38. 1 H NMR (300MHz, CD 3 OD) δ7.64 (d, 1H), 7.47 (s, 1H), 7.34 (d, 1H), 4.55 (s, 1H), 3.80 (s, 3H), 3.35 (s , 1H), 2.97 to 2.90 (m, 1H), 2.79 to 2.67 (m, 2H), 1.26 (s, 6H); MS (ESI): m/z 417.2 [M+H] + .
实施例39. 2-(7-(2-乙氧基丙基-2-基)-4,5,6,7氢噻吩并[2,3-c]哌啶-2-基)-6-氟-1H-苯并[d]咪唑-4-甲酰胺(I-39)：Example 39. 2-(7-(2-Ethoxypropyl-2-yl)-4,5,6,7-hydrothieno[2,3-c]piperidin-2-yl)-6- Fluor-1H-benzo[d]imidazole-4-carboxamide (I-39):
步骤1. 2-乙氧基-2-甲基-N-(2(噻吩-3-基)乙基)丙酰胺(39-1)Step 1. 2-Ethoxy-2-methyl-N-(2(thien-3-yl)ethyl)propanamide (39-1)
Figure PCTCN2016087368-appb-000200
Figure PCTCN2016087368-appb-000200
按照制备化合物37-1的操作方法，以3-噻吩乙胺(952.0mg,7.5mmol)和2-乙氧基-2-甲基丙酸(1.09g,0.83mmol)为原料制得1.2g油状化合物39-1，收率70％。MS(ESI):m/z 242.0[M+1]+。According to the procedure for the preparation of compound 37-1, 1-dithiophenethylamine (952.0 mg, 7.5 mmol) and 2-ethoxy-2-methylpropanoic acid (1.09 g, 0.83 mmol) were used as the starting material to obtain 1.2 g of oil. Compound 39-1, yield 70%. MS (ESI): m / z 242.0 [M + 1] +.
步骤2. 7-(2-乙氧基丙基-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)-羧酸叔丁酯(39-2)Step 2. 7-(2-Ethoxypropyl-2-yl)-4,5-dihydrothieno[2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (39-2 )
Figure PCTCN2016087368-appb-000201
Figure PCTCN2016087368-appb-000201
按照制备化合物14-3的操作方法，以2-乙氧基-2-甲基-N-(2(噻吩-3-基)乙基)丙酰胺(39-1)(964mg,4mmol)为原料制得360mg化合物39-2，收率27％。1H NMR(300MHz,CDCl3)δ7.17(d,J＝6.6Hz,1H),6.79(d,J＝6.6Hz,1H),5.41(s,0.5H),5.12(s,0.5H), 4.46(m,0.5H),4.24(m,0.5H),3.53(m,2H),3.13(m,1H),2.69(m,1H),2.60(s,1H),1.47(s,9H),1.35(s,3H),1.26(t,3H),1.07(d,J＝7.8Hz,3H)；MS(ESI):m/z 347.9[M+Na]+。Starting from 2-ethoxy-2-methyl-N-(2(thiophen-3-yl)ethyl)propanamide (39-1) (964 mg, 4 mmol) according to the procedure for the preparation of compound 14-3 360 mg of compound 39-2 was obtained in a yield of 27%. 1 H NMR (300MHz, CDCl 3 ) δ7.17 (d, J = 6.6Hz, 1H), 6.79 (d, J = 6.6Hz, 1H), 5.41 (s, 0.5H), 5.12 (s, 0.5H) , 4.46 (m, 0.5H), 4.24 (m, 0.5H), 3.53 (m, 2H), 3.13 (m, 1H), 2.69 (m, 1H), 2.60 (s, 1H), 1.47 (s, 9H) ), 1.35 (s, 3H), 1.26 (t, 3H), 1.07 (d, J = 7.8 Hz, 3H); MS (ESI): m/z 347.9 [M+Na] + .
步骤3. 2-溴-7-(乙氧基丙基-2-基)-4,5-二氢噻吩[2,3-c]吡啶-6(7H)-羧酸叔丁酯(39-3)Step 3. 2-Bromo-7-(ethoxypropyl-2-yl)-4,5-dihydrothiophene[2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (39- 3)
Figure PCTCN2016087368-appb-000202
Figure PCTCN2016087368-appb-000202
按照制备化合物12-3的制备方法,以7-(2-乙氧基丙基-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)-羧酸叔丁酯(39-2)(300mg,0.92mmol)为原料制得100mg化合物39-3。MS(ESI):m/z 425.7/427.8(1:1)[M+Na]+。According to the preparation method of the compound 12-3, 7-(2-ethoxypropyl-2-yl)-4,5-dihydrothieno[2,3-c]pyridine-6(7H)-carboxylate The tert-butyl acid ester (39-2) (300 mg, 0.92 mmol) was used as a starting material to obtain 100 mg of Compound 39-3. MS (ESI): m / z 425.7 / 427.8 (1: 1) [M + Na] +.
步骤4. 7-(2-乙氧基丙基-2-基)-2-甲酰基-4,5-二氢噻吩[2,3-c]吡啶-6(7H)-羧酸叔丁酯(39-4)Step 4. 7-(2-Ethoxypropyl-2-yl)-2-formyl-4,5-dihydrothiophene[2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (39-4)
Figure PCTCN2016087368-appb-000203
Figure PCTCN2016087368-appb-000203
按照制备化合物2-3的操作方法,以2-溴-7-(乙氧基丙基-2-基)-4,5-二氢噻吩[2,3-c]吡啶-6(7H)-羧酸叔丁酯(39-3)(100.0mg,0.25mmol)为原料制得38mg化合物39-4，收率43％。1H NMR(300MHz,CDCl3)δ9.82(s,1H),7.46(s,1H),5.41(s,0.5H),5.18(s,0.5H),4.48(dd,J＝3.3,10.2Hz,0.5H),4.26(dd,J＝2.1,10.2Hz,0.5H),3.51(q,J＝5.4Hz,2H),3.22～3.01(m,1H),2.76～2.58(m,2H),1.47(s,9H),1.34(s,3H),1.28(m,3H),1.05(s,3H)；MS(ESI):m/z 375.9[M+Na]+。According to the procedure for the preparation of compound 2-3, 2-bromo-7-(ethoxypropyl-2-yl)-4,5-dihydrothiophene[2,3-c]pyridine-6(7H)- Starting from tert-butyl carboxylate (39-3) (100.0 mg, 0.25 mmol), 38 mg of Compound 39-4 was obtained, yield 43%. 1 H NMR (300MHz, CDCl 3 ) δ9.82 (s, 1H), 7.46 (s, 1H), 5.41 (s, 0.5H), 5.18 (s, 0.5H), 4.48 (dd, J = 3.3,10.2 Hz, 0.5H), 4.26 (dd, J=2.1, 10.2 Hz, 0.5H), 3.51 (q, J = 5.4 Hz, 2H), 3.22 to 3.01 (m, 1H), 2.76 to 2.58 (m, 2H) , 1.47 (s, 9H), 1.34 (s, 3H), 1.28 (m, 3H), 1.05 (s, 3H); MS (ESI): m/z 375.9 [M+Na] + .
步骤5. 2-(4-氨甲酰基-6-氟-1H-苯并[d]咪唑-2-基)-7-(2-乙氧基丙基-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)-羧酸叔丁酯(39-5)Step 5. 2-(4-carbamoyl-6-fluoro-1H-benzo[d]imidazol-2-yl)-7-(2-ethoxypropyl-2-yl)-4,5- Dihydrothieno[2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (39-5)
Figure PCTCN2016087368-appb-000204
Figure PCTCN2016087368-appb-000204
按照制备化合物36-6的制备方法,以2,3-二氨基-5-氟苯甲酰胺(36-1)(20mg,0.12mmol)、化合物39-4(38mg,0.10mmol)为原料制得40mg化合物39-5，收率82.6％。1H NMR(300MHz,CD3OD)δ7.26(d,J＝8.4Hz,1H),7.46(s,1H),7.35(d,J＝6.3Hz),5.33(s,0.5H),5.22(s,0.5H),4.36(dd,J＝10.8Hz,1H),3.58(q,J＝5.1Hz,2H),3.23～3.12(m,1H),2.67(m,2H),1.5(s,9H),1.28(m,6H),1.13(s,3H)；MS(ESI):m/z 502.9[M+H]+。According to the preparation method of the compound 36-6, 2,3-diamino-5-fluorobenzamide (36-1) (20 mg, 0.12 mmol), and compound 39-4 (38 mg, 0.10 mmol) were used as raw materials. 40 mg of compound 39-5, yield 82.6%. 1 H NMR (300MHz, CD 3 OD) δ7.26 (d, J = 8.4Hz, 1H), 7.46 (s, 1H), 7.35 (d, J = 6.3Hz), 5.33 (s, 0.5H), 5.22 (s, 0.5H), 4.36 (dd, J = 10.8 Hz, 1H), 3.58 (q, J = 5.1 Hz, 2H), 3.23 to 3.12 (m, 1H), 2.67 (m, 2H), 1.5 (s) , 9H), 1.28 (m, 6H), 1.13 (s, 3H); MS (ESI): m/z 502.9 [M+H] + .
步骤6. 2-(7-(2-乙氧基丙基-2-基)-4,5,6,7氢噻吩并[2,3-c]哌啶-2-基)-6-氟-1H-苯并 [d]咪唑-4-甲酰胺(I-39)Step 6. 2-(7-(2-Ethoxypropyl-2-yl)-4,5,6,7-hydrothieno[2,3-c]piperidin-2-yl)-6-fluoro -1H-benzo [d]Imidazole-4-carboxamide (I-39)
Figure PCTCN2016087368-appb-000205
Figure PCTCN2016087368-appb-000205
按照制备化合物I-1的制备方法,以化合物39-5(40mg,0.08mmol)为原料制得16mg化合物1-39。1H NMR(300MHz,CD3OD)δ7.63(d,J＝7.2Hz,1H),7.48(s,1H),7.38(d,J＝5.1Hz,1H),4.15(s,1H),3.67～3.60(m,2H),3.40～3,38(m,1H),3,01～2.94(m,1H),2,84～2.73(m,2H),1.34(t,J＝5.4Hz,3H),1.26(s,3H),1,15(s,3H)；MS(ESI):m/z 403.2[M+H]+。According to the preparation method of the compound I-1, 16 mg of the compound 1-39 was obtained from the compound 39-5 (40 mg, 0.08 mmol). 1 H NMR (300MHz, CD 3 OD) δ7.63 (d, J = 7.2Hz, 1H), 7.48 (s, 1H), 7.38 (d, J = 5.1Hz, 1H), 4.15 (s, 1H), 3.67 to 3.60 (m, 2H), 3.40 to 3, 38 (m, 1H), 3, 01 to 2.94 (m, 1H), 2, 84 to 2.73 (m, 2H), 1.34 (t, J = 5.4 Hz) , 3H), 1.26 (s, 3H), 1, 15 (s, 3H); MS (ESI): m/z 403.2 [M+H] + .
实施例40. 6-氟-2-(7-(2-甲氧基丙基-2-基)-4,5,6,7氢噻吩并[2,3-c]哌啶-2-基)1H-苯并[d]咪唑-4-甲酰胺(I-40)Example 40. 6-Fluoro-2-(7-(2-methoxypropyl-2-yl)-4,5,6,7-hydrothieno[2,3-c]piperidin-2-yl ) 1H-benzo[d]imidazole-4-carboxamide (I-40)
步骤1：2-甲氧基-2-甲基-N-(2(噻吩-3-基)乙基)丙酰胺(40-1)Step 1: 2-Methoxy-2-methyl-N-(2(thien-3-yl)ethyl)propanamide (40-1)
Figure PCTCN2016087368-appb-000206
Figure PCTCN2016087368-appb-000206
按照制备化合物37-1的操作方法，以3-噻吩乙胺(605.0mg,4.8mmol)和2-甲氧基-2-甲基丙酸(562.0mmol,4.8mmol)为原料制的760.0mg油状化合物40-1，收率70％。MS(ESI):m/z 228.1[M+1]+。According to the procedure for the preparation of compound 37-1, 760.0 mg of oil was obtained from 3-thiophenethylamine (605.0 mg, 4.8 mmol) and 2-methoxy-2-methylpropanoic acid (562.0 mmol, 4.8 mmol). Compound 40-1, yield 70%. MS (ESI): m / z 228.1 [M + 1] +.
步骤2. 7-(2-甲氧基丙基-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)-羧酸叔丁酯(40-2)Step 2. 7-(2-Methoxypropyl-2-yl)-4,5-dihydrothieno[2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (40-2 )
Figure PCTCN2016087368-appb-000207
Figure PCTCN2016087368-appb-000207
按照制备化合物14-3的操作方法，以2-甲氧基-2-甲基-N-(2(噻吩-3-基)乙基)丙酰胺(40-1)(681mg,3mmol)为原料制得450mg化合物40-2，收率48％。1H NMR(300MHz,CDCl3)δ7.16(d,J＝10.5Hz,1H),6.79(d,J＝10.5Hz,1H),5.27(s,1H),4.50～4.20(m,1H),3.30(s,3H),3.23～3.03(m,1H),2.75～2.55(m,2H),1.47(s,9H),1.35(s,3H),1.08(s,3H)；MS(ESI):m/z 333.9[M+Na]+。According to the procedure for the preparation of compound 14-3, 2-methoxy-2-methyl-N-(2(thiophen-3-yl)ethyl)propanamide (40-1) (681 mg, 3 mmol) was used. 450 mg of compound 40-2 was obtained in a yield of 48%. 1 H NMR (300MHz, CDCl 3 ) δ 7.16 (d, J = 10.5 Hz, 1H), 6.79 (d, J = 10.5 Hz, 1H), 5.27 (s, 1H), 4.50 to 4.20 (m, 1H) , 3.30 (s, 3H), 3.23 to 3.03 (m, 1H), 2.75 to 2.55 (m, 2H), 1.47 (s, 9H), 1.35 (s, 3H), 1.08 (s, 3H); MS (ESI) ): m/z 333.9 [M+Na] + .
步骤3. 2-溴-7-(甲氧基丙基-2-基)-4,5-二氢噻吩[2,3-c]吡啶-6(7H)-羧酸叔丁酯(40-3)Step 3. 2-Bromo-7-(methoxypropyl-2-yl)-4,5-dihydrothiophene[2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (40- 3)
Figure PCTCN2016087368-appb-000208
Figure PCTCN2016087368-appb-000208
按照制备化合物12-3的制备方法,以7-(2-乙氧基丙基-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)-羧酸叔丁酯(40-2)(440mg,14mmol)为原料制得400mg化合物40-3。MS(ESI):m/z 411.7/413.7(1:1)[M+Na]+。According to the preparation method of the compound 12-3, 7-(2-ethoxypropyl-2-yl)-4,5-dihydrothieno[2,3-c]pyridine-6(7H)-carboxylate The acid tert-butyl ester (40-2) (440 mg, 14 mmol) was used as a starting material to obtain 400 mg of Compound 40-3. MS (ESI): m / z 411.7 / 413.7 (1: 1) [M + Na] +.
步骤4. 2-甲酰基-7-(2-甲氧基丙基-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)羧酸叔丁酯(40-4)Step 4. 2-Formyl-7-(2-methoxypropyl-2-yl)-4,5-dihydrothieno[2,3-c]pyridine-6(7H)carboxylic acid tert-butyl ester (40-4)
Figure PCTCN2016087368-appb-000209
Figure PCTCN2016087368-appb-000209
按照制备化合物2-3的操作方法,以2-溴-7-(甲氧基丙基-2-基)-4,5-二氢噻吩[2,3-c]吡啶-6(7H)-羧酸叔丁酯(40-3)(850.0mg,2.3mmol)为原料制得493mg化合物40-4，收率67％。1H NMR(300MHz,CDCl3)δ9.84(s,1H),7.46(s,1H),5.30(s,1H),4.52～4.25(m,1H),3.39(s,3H),3.23～3.02(m,1H),2.72～2.60(m,2H),1.48(s,9H),1.36(s,3H),1.07(s,3H)；MS(ESI):m/z 361.9[M+Na]+。According to the procedure for the preparation of compound 2-3, 2-bromo-7-(methoxypropyl-2-yl)-4,5-dihydrothiophene[2,3-c]pyridine-6(7H)- Starting from tert-butyl carboxylate (40-3) (850.0 mg, 2.3 mmol), 493 mg of Compound 40-4 was obtained. 1 H NMR (300MHz, CDCl 3 ) δ 9.84 (s, 1H), 7.46 (s, 1H), 5.30 (s, 1H), 4.52 to 4.25 (m, 1H), 3.39 (s, 3H), 3.23 3.02 (m, 1H), 2.72 to 2.60 (m, 2H), 1.48 (s, 9H), 1.36 (s, 3H), 1.07 (s, 3H); MS (ESI): m/z 361.9 [M+Na ] + .
步骤5. 2-(4-氨甲酰基-6-氟-1H-苯并[d]咪唑-2基)-7-(2-甲氧基丙基-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)-羧酸叔丁酯(40-5)Step 5. 2-(4-carbamoyl-6-fluoro-1H-benzo[d]imidazol-2-yl)-7-(2-methoxypropyl-2-yl)-4,5-di Hydrothieno[2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (40-5)
Figure PCTCN2016087368-appb-000210
Figure PCTCN2016087368-appb-000210
按照制备化合物36-6的制备方法,以2,3-二氨基-5-氟苯甲酰胺(36-1)(23.6mg,0.14mmol)和化合物40-4(45.0mg,0.13mmol)为原料制得55mg化合物40-5，收率84.8％。1H NMR(300MHz,CD3OD)δ7.54(d,1H),7.37(s,1H),7.24(d,1H),5.18(s,1H),4.33～4.19(m,1H),3.25(s,3H),3.18～3.05(m,1H),2.58(d,2H),1.40(s,9H),1.28(s,3H),1.04(s,3H)；MS(ESI):m/z 489.2[M+H]+。According to the preparation method of the compound 36-6, 2,3-diamino-5-fluorobenzamide (36-1) (23.6 mg, 0.14 mmol) and the compound 40-4 (45.0 mg, 0.13 mmol) were used as the starting materials. 55 mg of compound 40-5 was obtained in a yield of 84.8%. 1 H NMR (300 MHz, CD 3 OD) δ 7.54 (d, 1H), 7.37 (s, 1H), 7.24 (d, 1H), 5.18 (s, 1H), 4.33 to 4.19 (m, 1H), 3.25 (s, 3H), 3.18 to 3.05 (m, 1H), 2.58 (d, 2H), 1.40 (s, 9H), 1.28 (s, 3H), 1.04 (s, 3H); MS (ESI): m/ z 489.2[M+H] + .
步骤6. 6-氟-2-(7-(2-甲氧基异丙-2-基)-4,5,6,7氢噻吩并[2,3-c]哌啶-2-基)1H-苯并[d]咪唑-4-甲酰胺(I-40)Step 6. 6-Fluoro-2-(7-(2-methoxyisopropyl-2-yl)-4,5,6,7-hydrothieno[2,3-c]piperidin-2-yl) 1H-benzo[d]imidazole-4-carboxamide (I-40)
Figure PCTCN2016087368-appb-000211
Figure PCTCN2016087368-appb-000211
将2-(4-氨甲酰基-6-氟-1H-苯并[d]咪唑-2基)-7-(2-甲氧基异丙-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)-羧酸叔丁酯(40-5)(30.0mg,0.06mmol)和三氟乙酸(0.5mL)加入到二氯甲烷(20.0mL)中于室温下搅拌直到原料反应完全。将反应体系浓缩的粗产品.粗产品经柱层析(二氯甲烷：甲醇＝20：1)得18mg化合物I-40.1H NMR(300MHz,CD3OD) δ7.63(d,1H),7.49(s,1H),7.37(d,1H),4.06(s,1H),3.39(s,3H),3.35～3.33(m,1H),2.95～2.88(m,1H),2.80～2.64(m,2H),1.36(s 3H),1.10(s,3H)；MS(ESI):m/z 389.2[M+H]+ 2-(4-carbamoyl-6-fluoro-1H-benzo[d]imidazol-2-yl)-7-(2-methoxyisopropyl-2-yl)-4,5-dihydrothiophene [2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (40-5) (30.0 mg, 0.06 mmol) and trifluoroacetic acid (0.5 mL) were added to dichloromethane (20.0 mL) Stir at room temperature until the starting material is completely reacted. The crude product was concentrated. The crude product was purified by column chromatography (dichloromethanol:methanol = 20:1) to give 18 mg of compound I-40. 1 H NMR (300 MHz, CD 3 OD) δ 7.63 (d, 1H) , 7.49 (s, 1H), 7.37 (d, 1H), 4.06 (s, 1H), 3.39 (s, 3H), 3.35 to 3.33 (m, 1H), 2.95 to 2.88 (m, 1H), 2.80 to 2.64 (m, 2H), 1.36 (s 3H), 1.10 (s, 3H); MS (ESI): m/z 389.2 [M+H] +
实施例41. 6-氟-2-(7-(2-甲氧基异丙-2-基)-4,5,6,7-四氢噻吩[2,3-c]哌啶-2-基)-1-甲基-1H-苯并[d]咪唑-4-甲酰胺(I-41)：Example 41. 6-Fluoro-2-(7-(2-methoxyisopropyl-2-yl)-4,5,6,7-tetrahydrothiophene[2,3-c]piperidin-2- 1-methyl-1H-benzo[d]imidazole-4-carboxamide (I-41):
步骤1：2-(4-氨基甲酰基-6-氟-1-甲基-1H-苯并[d]咪唑-2-基)-7-(2-甲氧基异丙-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)-羧酸叔丁酯(41-1)和2-(6-氟-1-甲基-4(甲胺甲酰基)-1H-苯并[d]咪唑-2-基)-7-(2-甲氧基异丙-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)-羧酸叔丁酯(41-2)Step 1: 2-(4-carbamoyl-6-fluoro-1-methyl-1H-benzo[d]imidazol-2-yl)-7-(2-methoxyisopropyl-2-yl) -4,5-dihydrothieno[2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (41-1) and 2-(6-fluoro-1-methyl-4(methylamine) Formyl)-1H-benzo[d]imidazol-2-yl)-7-(2-methoxyisopropyl-2-yl)-4,5-dihydrothieno[2,3-c]pyridine -6(7H)-tert-butyl carboxylic acid (41-2)
Figure PCTCN2016087368-appb-000212
Figure PCTCN2016087368-appb-000212
氮气保护下,将2-(4-氨甲酰基-6-氟-1H-苯并[d]咪唑-2基)-7-(2-甲氧基异丙-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)-羧酸叔丁酯(40-5)(55.0mg,0.11mmol)溶于无水DMF(8.0mL)中.随后冷却到0℃左右缓慢加入氢化钠(60％W/W,7.2mg,0.18mmol).加入完毕,升温到40℃左右搅拌1h。随后冷却到室温缓慢加入碘甲烷(38mg,0.28mmol)于室温下搅拌1小时原料反应物完全.将反应体系倒入到冰水中,用二氯甲烷萃取得到有机相，有机相干燥,浓缩得到粗品。粗产品经柱层析(二氯甲烷：甲醇＝50：1)得12mg化合物41-1和24mg化合物41-2.2-(4-carbamoyl-6-fluoro-1H-benzo[d]imidazol-2-yl)-7-(2-methoxyisopropyl-2-yl)-4,5 under nitrogen -Dihydrothieno[2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (40-5) (55.0 mg, 0.11 mmol) dissolved in anhydrous DMF (8.0 mL). Sodium hydride (60% W/W, 7.2 mg, 0.18 mmol) was slowly added at about 0 ° C. After the addition was completed, the temperature was raised to about 40 ° C and stirred for 1 h. After cooling to room temperature, iodomethane (38 mg, 0.28 mmol) was slowly added and stirred at room temperature for 1 hour. The reaction mixture was completed. The reaction mixture was poured into ice water and extracted with dichloromethane to give an organic phase. . The crude product was subjected to column chromatography (dichloromethane:methanol = 50:1) to give 12 mg of compound 41-1 and 24 mg of compound 41-2.
化合物41-1：1H NMR(300MHz,CD3OD)7.67(dd,1H),7.46(s,1H),7.44(dd,1H),5.36(brs,0.5H),5.24(brs,0.5H),4.37(m,1H),3.99(s,3H),3.34(s,3H),3.31(m,1H),2.71(m,2H),1.52(s,9H),1.39(s,3H),1.15(s,3H).MS(ESI):m/z 503.2[M+H]+.Compound 41-1: 1 H NMR (300 MHz, CD 3 OD) 7.67 (dd, 1H), 7.46 (s, 1H), 7.44 (dd, 1H), 5.36 (brs, 0.5H), 5.24 (brs, 0.5H) ), 4.37 (m, 1H), 3.99 (s, 3H), 3.34 (s, 3H), 3.31 (m, 1H), 2.71 (m, 2H), 1.52 (s, 9H), 1.39 (s, 3H) , 1.15 (s, 3H). MS (ESI): m/z 503.2 [M+H] + .
化合物41-2：1H NMR(300MHz,CD3OD)7.46(dd,1H),7.30(d,1H),7.21(dd,1H),5.33(s,0.5H),5.23(s,0.5H),4.35(m,1H),3.75(s,3H),3.35(m,1H),3.33(s,3H),2.96(s,3H),2.67(d,2H),1.52(s,9H),1.38(s,3H),1.13(s,3H).MS(ESI):m/z 517.3[M+H]+.Compound 41-2: 1 H NMR (300MHz, CD 3 OD) 7.46 (dd, 1H), 7.30 (d, 1H), 7.21. (dd, 1H), 5.33 (s, 0.5H), 5.23 (s, 0.5H) ), 4.35 (m, 1H), 3.75 (s, 3H), 3.35 (m, 1H), 3.33 (s, 3H), 2.96 (s, 3H), 2.67 (d, 2H), 1.52 (s, 9H) , 1.38 (s, 3H), 1.13 (s, 3H). MS (ESI): m/z 517.3 [M+H] + .
步骤2. 6-氟-2-(7-(2-甲氧基异丙-2-基)-4,5,6,7-四氢噻吩[2,3-c]哌啶-2-基)-1-甲基苯-1-H并[d]咪唑-4-甲酰胺(I-41)Step 2. 6-Fluoro-2-(7-(2-methoxyisopropyl-2-yl)-4,5,6,7-tetrahydrothiophene[2,3-c]piperidin-2-yl )-1-methylphenyl-1-H-[d]imidazole-4-carboxamide (I-41)
Figure PCTCN2016087368-appb-000213
Figure PCTCN2016087368-appb-000213
将2-(4-氨基甲酰基-6-氟-1-甲基-1H-苯并[d]咪唑-2-基)-7-(2-甲氧基异丙-2- 基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)-羧酸叔丁酯(41-1)(12.0mg,0.023mmol)溶于乙酸乙酯(20ml)中,随后在0℃左右充入HCl(gas)0.5h.于室温下反应过夜.加入饱和碳酸氢钠水溶液调PH到中性。静置、分液得到有机相。干燥、浓缩得到粗品，粗产品经柱层析(二氯甲烷：甲醇＝20：1)得4.4mg I-41。1H NMR(300MHz,CD3OD)7.71(dd,1H),7.63(s,1H),7.10(dd,1H),5.37(s,1H),4.10(s,1H),4.01(s,3H),3.37(s,3H),2.96(m,1H),2.73(m,2H),1.36(s,3H),1.12(s,3H)。MS(ESI):m/z 403.2[M+H]+。2-(4-carbamoyl-6-fluoro-1-methyl-1H-benzo[d]imidazol-2-yl)-7-(2-methoxyisopropyl-2-yl)-4 , 5-dihydrothieno[2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (41-1) (12.0 mg, 0.023 mmol) dissolved in ethyl acetate (20 ml) HCl (gas) was added at about 0 ° C for 0.5 h. The reaction was allowed to proceed overnight at room temperature. A saturated aqueous sodium hydrogencarbonate solution was added to adjust the pH to neutral. The organic phase was obtained by standing and liquid separation. Drying and concentrating gave a crude material. EtOAc m. 1 H NMR (300MHz, CD 3 OD) 7.71 (dd, 1H), 7.63 (s, 1H), 7.10 (dd, 1H), 5.37 (s, 1H), 4.10 (s, 1H), 4.01 (s, 3H ), 3.37 (s, 3H), 2.96 (m, 1H), 2.73 (m, 2H), 1.36 (s, 3H), 1.12 (s, 3H). MS (ESI): m / z 403.2 [M + H] +.
实施例42. 6-氟-2-(7-(2-2-甲氧基异丙-2-基)-4,5,6,7-四氢噻吩[2,3-c]哌啶-2-基)-N,1-二甲基-1-H并[d]咪唑-4-甲酰胺(I-42)Example 42. 6-Fluoro-2-(7-(2-2-methoxyisopropyl-2-yl)-4,5,6,7-tetrahydrothiophene [2,3-c]piperidine- 2-yl)-N,1-dimethyl-1-H-[d]imidazole-4-carboxamide (I-42)
Figure PCTCN2016087368-appb-000214
Figure PCTCN2016087368-appb-000214
按实施例41步骤2的方法制备I-42：以24mg,0.046mmol的2-(6-氟-1-甲基-4(甲胺甲酰基)-1H-苯并[d]咪唑-2-基)-7-(2-甲氧基异丙-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)-羧酸叔丁酯(41-2)为原料得到12.6mg化合物I-42。1H NMR(300MHz,CD3OD)7.65(dd,1H),7.40(s,1H),7.36(dd,1H),4.10(s,1H),3.96(s,3H),3.38(s,3H),3.35(m,1H),3.07(s,3H),2.96(m,1H),2.74(m,2H),1.37(s,3H),1.14(s,3H)；MS(ESI):m/z 417.2[M+H]+.1-42 was prepared as in Step 2 of Example 41: 24 mg, 0.046 mmol of 2-(6-fluoro-1-methyl-4(methylcarbamoyl)-1H-benzo[d]imidazole-2- -7-(2-Methoxyisopropyl-2-yl)-4,5-dihydrothieno[2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (41-2 ) 12.6 mg of compound I-42 was obtained as a starting material. 1 H NMR (300MHz, CD 3 OD) 7.65 (dd, 1H), 7.40 (s, 1H), 7.36 (dd, 1H), 4.10 (s, 1H), 3.96 (s, 3H), 3.38 (s, 3H ), 3.35 (m, 1H), 3.07 (s, 3H), 2.96 (m, 1H), 2.74 (m, 2H), 1.37 (s, 3H), 1.14 (s, 3H); MS (ESI): m /z 417.2[M+H] + .
实施例43. 1-二氟甲基-6-氟-2(7-(2-甲氧基异丙-2-基)-4,5,6,7-四氢噻吩[2,3-c]哌啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-43)Example 43. 1-Difluoromethyl-6-fluoro-2(7-(2-methoxyisopropyl-2-yl)-4,5,6,7-tetrahydrothiophene [2,3-c ]piperidin-2-yl)-1H-benzo[d]imidazole-4-carboxamide (I-43)
步骤1：2-(4-甲酰胺基-1-二氟甲基)-6-氟-1H-苯并[d]咪唑-2-基)-7-(2-甲氧基异丙-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)-羧酸叔丁酯(43-1)Step 1: 2-(4-Carboxamido-1-difluoromethyl)-6-fluoro-1H-benzo[d]imidazol-2-yl)-7-(2-methoxyisopropyl-2 -yl)-4,5-dihydrothieno[2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (43-1)
Figure PCTCN2016087368-appb-000215
Figure PCTCN2016087368-appb-000215
将2-(4-氨甲酰基-6-氟-1H-苯并[d]咪唑-2基)-7-(2-甲氧基异丙-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)-羧酸叔丁酯(40-5)(40.0mg,0.082mmol)、二氟氯乙酸钠(25.0mg，0.164mmol)和碳酸钾(23.0mg,0.164mmol)加入到N,N-二甲基甲酰胺(10.0ml)中。随后升温到90℃下反应直到主原料反应完全。冷却，将反应体系倒入到冰水中，用乙酸乙酯萃取得乙酸乙酯相。干燥，浓缩的粗品。粗产品经柱层析(石油醚：乙酸乙酯＝4：1)得32mg化合物43-1。1H NMR(300MHz,CD3OD)δ8.73(s,1H),7.79(m,1H),7.71(d,1H),7.38(s,1H),5.37(d,0.5H),4.77(s,0.5H),4.58(s,2H),3.34(s,3H),2.72(m,2H),1.50(s,9H),1.38(d,3H),1.15(s,3H).MS(ESI):m/z 539.1[M+H]+. 2-(4-carbamoyl-6-fluoro-1H-benzo[d]imidazol-2-yl)-7-(2-methoxyisopropyl-2-yl)-4,5-dihydrothiophene And [2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (40-5) (40.0 mg, 0.082 mmol), sodium difluorochloroacetate (25.0 mg, 0.164 mmol) and potassium carbonate (23.0) Mg, 0.164 mmol) was added to N,N-dimethylformamide (10.0 mL). The temperature is then raised to 90 ° C until the main starting material is completely reacted. After cooling, the reaction system was poured into ice water and extracted with ethyl acetate to give ethyl acetate. Dry, concentrated crude. The crude product was purified by column chromatography (EtOAc:EtOAc:EtOAc 1 H NMR (300MHz, CD 3 OD) δ8.73 (s, 1H), 7.79 (m, 1H), 7.71 (d, 1H), 7.38 (s, 1H), 5.37 (d, 0.5H), 4.77 ( s, 0.5H), 4.58 (s, 2H), 3.34 (s, 3H), 2.72 (m, 2H), 1.50 (s, 9H), 1.38 (d, 3H), 1.15 (s, 3H). ESI): m/z 539.1 [M+H] + .
步骤2. 1-二氟甲基-6-氟-2(7-(2-甲氧基异丙-2-基)-4,5,6,7-四氢噻吩[2,3-c]哌啶-2-基)-1H-苯并[d]咪唑-4-甲酰胺(I-43)Step 2. 1-Difluoromethyl-6-fluoro-2(7-(2-methoxyisopropyl-2-yl)-4,5,6,7-tetrahydrothiophene [2,3-c] Piperidin-2-yl)-1H-benzo[d]imidazole-4-carboxamide (I-43)
Figure PCTCN2016087368-appb-000216
Figure PCTCN2016087368-appb-000216
将2-(4-甲酰胺基-1-二氟甲基)-6-氟-1H-苯并[d]咪唑-2-基)-7-(2-甲氧基异丙-2-基)-4,5-二氢噻吩并[2,3-c]吡啶-6(7H)-羧酸叔丁酯(43-1)(32.0mg,0.06mmol)加入到乙腈(10.0mL)中溶解并冷却到0℃，随后加入三甲基碘硅烷(23.8mg,0.12mmoL)反应知道原料完全反应。将反应体系倒入到冰水中，随后用乙酸乙酯进行萃取得乙酸乙酯相。干燥，浓缩的粗品。粗产品经柱层析(二氯甲烷：甲醇＝20：1)得10.0mg白色固体I-43.1H NMR(300MHz,d6-DMSO)δ8.79(s,1H,NH),8.16(s,1H),7.85(d,1H),7.71(d,1H),7.43(s,1H),3.95(s,1H),3.27(s,3H),3.17(m,2H),2.63(m,2H),1.26(s,3H),1.00(s,3H)；MS(ESI):m/z439.1[M+H]+。2-(4-Carboxamido-1-difluoromethyl)-6-fluoro-1H-benzo[d]imidazol-2-yl)-7-(2-methoxyisopropyl-2-yl -4,5-Dihydrothieno[2,3-c]pyridine-6(7H)-carboxylic acid tert-butyl ester (43-1) (32.0 mg, 0.06 mmol) was dissolved in acetonitrile (10.0 mL) The mixture was cooled to 0 ° C, followed by the addition of trimethylsilyl iodide (23.8 mg, 0.12 mmol) to know the complete reaction of the starting material. The reaction system was poured into ice water, followed by extraction with ethyl acetate to give an ethyl acetate phase. Dry, concentrated crude. The crude product was purified by column chromatography. (Dichloromethane: methanol = 20: 1) to give 10.0mg as a white solid I-43 1 H NMR (300MHz , d 6 -DMSO) δ8.79 (s, 1H, NH), 8.16 ( s, 1H), 7.85 (d, 1H), 7.71 (d, 1H), 7.43 (s, 1H), 3.95 (s, 1H), 3.27 (s, 3H), 3.17 (m, 2H), 2.63 (m) , 2H), 1.26 (s, 3H), 1.00 (s, 3H); MS (ESI): m/z 439.1 [M+H] + .
实施例44. (+)-6-氟-2-(7-(2-甲氧基异丙-2-基)-4,5,6,7氢噻吩并[2,3-c]哌啶-2-基)1H-苯并[d]咪唑-4-甲酰胺(I-44)和(-)-6-氟-2-(7-(2-甲氧基异丙-2-基)-4,5,6,7氢噻吩并[2,3-c]哌啶-2-基)1H-苯并[d]咪唑-4-甲酰胺(I-45)Example 44. (+)-6-Fluoro-2-(7-(2-methoxyisopropyl-2-yl)-4,5,6,7-hydrothieno[2,3-c]piperidine -2-yl)1H-benzo[d]imidazole-4-carboxamide (I-44) and (-)-6-fluoro-2-(7-(2-methoxyisopropyl-2-yl) -4,5,6,7-hydrothieno[2,3-c]piperidin-2-yl)1H-benzo[d]imidazole-4-carboxamide (I-45)
将消旋体6-氟-2-(7-(2-甲氧基异丙-2-基)-4,5,6,7氢噻吩并[2,3-c]哌啶-2-基)1H-苯并[d]咪唑-4-甲酰胺(I-40)(140mg,0.36mmol)经手性制备液相分离得到60.0mg化合物(I-44))和60.0mg化合物(I-45)。分离条件为：色谱柱型号：Superchiral S-OZ；色谱柱规格：0.40cm I.D.×25cm L；进样量：5uL；流动相：CO2/MeOH/DEA＝60/40/0.05(v/v/v)；流速：2.4ml/min；检测条件：UVλ＝220nm；柱温：35℃。The racemate 6-fluoro-2-(7-(2-methoxyisopropyl-2-yl)-4,5,6,7-hydrothieno[2,3-c]piperidin-2-yl 1H-benzo[d]imidazole-4-carboxamide (I-40) (140 mg, 0.36 mmol) was isolated by chiral preparative liquid phase to give 60.0 mg of compound (I-44)) and 60.0 mg of compound (I-45) . The separation conditions were: column type: Superchiral S-OZ; column size: 0.40 cm ID × 25 cm L; injection amount: 5 uL; mobile phase: CO 2 / MeOH / DEA = 60 / 40 / 0.05 (v / v / v); flow rate: 2.4 ml/min; detection conditions: UV λ = 220 nm; column temperature: 35 ° C.
I-44：1H NMR(300MHz,CDCl3)δ7.41(s,1H),7.26(s,2H),4.07(s,1H),3.50(s,1H),3.36(s,3H),3.00(s,1H),2.67(m,2H),1.32(s,3H),1.19(s,3H)；MS(ESI)m/z:389.2[M+H]+。[α]20D+1.675(c,2.042,MeOH)；ee>99％；tR＝4.12min,I-44: 1 H NMR (300MHz, CDCl 3 ) δ 7.41 (s, 1H), 7.26 (s, 2H), 4.07 (s, 1H), 3.50 (s, 1H), 3.36 (s, 3H), 3.00 (s, 1H), 2.67 (m, 2H), 1.32 (s, 3H), 1.19 (s, 3H); MS (ESI) m/z: 389.2 [M+H] + . [α]20D+1.675 (c, 2.042, MeOH); ee >99%; tR=4.12 min,
I-45：1H NMR(300MHz,CDCl3)δ7.47(s,1H),7.26(s,2H),4.05(s,1H),3.49(s,1H),3.36(s,3H),2.99(s,1H),2.55～2.68(m,2H),1.34(s,3H),1.08(s,3H)；MS(ESI):m/z389.2[M+H]+。[α]20D-1.560(c,2.024,MeOH)；ee>99％；tR＝5.38min。I-45: 1 H NMR (300MHz, CDCl 3 ) δ 7.47 (s, 1H), 7.26 (s, 2H), 4.05 (s, 1H), 3.49 (s, 1H), 3.36 (s, 3H), 2.99 (s, 1H), 2.55 to 2.68 (m, 2H), 1.34 (s, 3H), 1.08 (s, 3H); MS (ESI): m/z 389.2 [M+H] + . [α] 20D-1.560 (c, 2.024, MeOH); ee >99%; tR = 5.38 min.
活性测试试验Activity test
分子水平PARP酶活性抑制实验Molecular level PARP enzyme activity inhibition experiment
实验方法：酶联免疫吸附法(Enzyme-Linked Immunosorbent Assay,ELISA)(参考文献：Decker,P.；Miranda,E.A.；de Murcia,G.and Muller,S.An improved nonisotopic test to screen a large series of new inhibitor molecules of poly(ADP-ribose)polymerase activity  for therapeutic applications.Clin.Cancer Res.1999,5,1169-1172.)。其原理是将底物组蛋白包被在吸附性的96孔板上，加入PARP1/2重组酶、底物NAD+、激活的DNA使PARP1/2发生酶反应，PARP酶可以催化切割底物DAD+的酰胺键，生成烟酰胺和ADP核糖，并以ADP核糖为底物在靶蛋白(Histone)上合成聚腺苷二磷酸核糖(PAR)，然后加入抗PAR(anti-PAR)的抗体，检测96孔板上所包被的组蛋白上的产物PAR的强度，就可以反映出PARP酶活性。Experimental method: Enzyme-Linked Immunosorbent Assay (ELISA) (Reference: Decker, P.; Miranda, EA; de Murcia, G. and Muller, S. An improved nonisotopic test to screen a large series of New inhibitor molecules of poly(ADP-ribose) polymerase activity for therapeutic applications. Clin. Cancer Res. 1999, 5, 1169-1172.). The principle is to coat the substrate histone in an adsorption 96-well plate, add PARP1/2 recombinase, substrate NAD + , activated DNA to enzymatically react PARP1/2, PARP enzyme can catalyze the cleavage of substrate DAD + amide bond to form nicotinamide and ADP ribose, and synthesize poly(ADP) ribose (PAR) on target protein (Histone) with ADP ribose as substrate, then add anti-PAR (anti-PAR) antibody, detect The intensity of the product PAR on the histones coated on the 96-well plate reflects the PARP enzyme activity.
具体方法如下：The specific method is as follows:
1.组蛋白是PARP公认的重要底物。将Histone(1ng/mL)其用无钾离子的PBS(10mM磷酸钠缓冲液，150mM NaCl,pH 7.2-7.4)包被96孔酶标板,置37℃摇床包被过夜；弃去孔中液体。用120μL/孔的T-PBS(含0.1％Tween-20的PBS)洗板5次，于37℃烘箱中干燥。1. Histone is an important substrate recognized by PARP. Histone (1 ng/mL) was coated with 96-well microtiter plate with potassium-free PBS (10 mM sodium phosphate buffer, 150 mM NaCl, pH 7.2-7.4), and coated overnight at 37 ° C; discarded in the well liquid. The plate was washed 5 times with 120 μL/well of T-PBS (PBS containing 0.1% Tween-20), and dried in an oven at 37 °C.
2.加入NAD+(终浓度8μM)，DNA(100ng/孔)，PARP1(10ng/孔)(用PARP1酶反应缓冲液稀释，缓冲液含50mM Tris(三羟甲基氨基甲烷)，2mM MgCl2,pH 8.0)，每孔加入一定稀释浓度(终浓度从10μM开始，10倍稀释6个梯度)10μL的抑制剂(阳性对照化合物为：AZD2281，商品名Olaparib，购自LC，Laboratories公司；ABT-888，商品名Veliparib，购自上海瀚香生物科技有限公司；2-(4-羟基苯基)-1H-苯并[d]咪唑-4-甲酰胺，编号为D-1，参照文献方法制备(参考文献：White,A.W.；Almassy,R.；Calvert,A.H.；Curtin,N.J.；Griffin,R.J.；Hostomsky,Z.；Maegley,K.；Newell,D.R.；Srinivasan,S.and Golding,B.T.Resistance-modifying agents.9.Synthesis and biological properties of benzimidazole inhibitors of the DNA repair enzyme poly(ADP-ribose)polymerase.J.Med.Chem.2000,43,4084-4097.)，1H NMR(300MHz,Methanol-d4)δ8.03(d,J＝8.7Hz,2H),7.89(dd,J＝7.6,1.1Hz,1H),7.68(dd,J＝8.1,1.1Hz,1H),7.29(dd,J＝8.3,7.4Hz,1H),6.94(d,J＝8.7Hz,2H)；MS(ESI)254[M+H]+。)，每个浓度设置2个重复，反应体系共100μL/孔(用前述PARP1酶反应缓冲液补足)，37℃摇床反应1h，设置空白、阳性、阴性对照(以只加反应缓冲稀释液的为空白对照孔，不加抑制剂加酶孔为阳性对照孔，以不加抑制剂的为阴性对照孔)。启动反应，置37℃摇床反应1小时。2. Add NAD + (final concentration 8 μM), DNA (100 ng/well), PARP1 (10 ng/well) (diluted with PARP1 enzyme reaction buffer, 50 mM Tris (tris), 2 mM MgCl 2 , pH 8.0), add a certain dilution concentration per well (final concentration from 10 μM, 10 times dilution of 6 gradients) 10 μL of inhibitor (positive control compound: AZD2281, trade name Olaparib, purchased from LC, Laboratories; ABT- 888, trade name Veliparib, purchased from Shanghai Muxiang Biotechnology Co., Ltd.; 2-(4-hydroxyphenyl)-1H-benzo[d]imidazole-4-carboxamide, numbered D-1, prepared by reference method (References: White, AW; Almassy, R.; Calvert, AH; Curtin, NJ; Griffin, RJ; Hostomsky, Z.; Maegley, K.; Newell, DR; Srinivasan, S. and Golding, BT Resistance-modifying agents .9.Synthesis and biological properties of benzimidazole inhibitors of the DNA repair enzyme poly(ADP-ribose)polymerase.J.Med.Chem.2000,43,4084-4097.), 1 H NMR (300MHz, Methanol-d 4 ) δ 8.03 (d, J = 8.7 Hz, 2H), 7.89 (dd, J = 7.6, 1.1 Hz, 1H), 7.68 (dd, J = 8.1, 1.1 Hz, 1H), 7.29 (dd, J = 8.3, 7.4 Hz, 1H), 6.94 (d, J = 8.7 Hz, 2H) MS (ESI) 254 [M+H] + .), 2 replicates per concentration, 100 μL/well of the reaction system (complemented with the aforementioned PARP1 enzyme reaction buffer), shaken at 37 ° C for 1 h, set blank, Positive and negative controls (only blank control wells with reaction buffer dilution, no inhibitor plus enzyme wells as positive control wells, negative control wells without inhibitor). The reaction was started and allowed to react at 37 ° C for 1 hour.
3.用PBS-T洗板三遍，加入一抗Anti-PAR Polyclonal Antibody(Rabbit)(1:6000,用含5mg/mL BSA的PBS-T稀释)，100μL/孔,37℃摇床孵育1h；3. Wash the plate three times with PBS-T, add anti-Anti-PAR Polyclonal Antibody (Rabbit) (1:6000, diluted with PBS-T containing 5 mg/mL BSA), 100 μL/well, incubate for 1 h at 37 ° C shaker. ;
4.用PBS-T洗板三遍,加入过氧化物酶标记的二抗(抗兔抗体)(1:2000,用含5μg/mL BSA的PBS-T稀释)，100μL/孔,37℃摇床反应30分钟；4. Wash the plate three times with PBS-T, add peroxidase-labeled secondary antibody (anti-rabbit antibody) (1:2000, diluted with PBS-T containing 5 μg/mL BSA), 100 μL/well, shake at 37 °C Bed reaction for 30 minutes;
5.加入2mg/mL的OPD(O-Phenylenediamine Dihydrochloride)显色液100μL/孔(用含有0.1％H2O2的0.1M柠檬酸－柠檬酸钠缓冲液(pH＝5.4)稀释)，25℃避光反应15分钟(OPD溶解时需用超声，显色液需现配现用)。5. Add 2 mg/mL OPD (O-Phenylenediamine Dihydrochloride) coloring solution 100 μL/well (diluted with 0.1 M citric acid-sodium citrate buffer (pH=5.4) containing 0.1% H 2 O 2 ), 25 ° C The reaction is protected from light for 15 minutes (ultrasound is required for the dissolution of OPD, and the coloring solution needs to be used now).
6.用50μL/孔的2M H2SO4终止反应，用Molecular Devices酶标仪(购自美国分子仪器公司(MDC)，型号为SpectraMax 190Microplate Reader(90V to 240V))读数，波长490nm测OD值。 6. Stop the reaction with 50 μL/well of 2M H 2 SO 4 and read with a Molecular Devices microplate reader (available from Molecular Instruments, Inc., model SpectraMax 190 Microplate Reader (90V to 240V)). OD at 490 nm. .
按照以下公式计算药物对PARP1酶活性的抑制程度：The degree of inhibition of PARP1 enzyme activity by the drug was calculated according to the following formula:
抑制率(％)＝[(OD阳性对照孔-OD阴性对照孔)-(OD给药孔-OD阴性对照孔)]/(OD阳性对照孔-OD阴性对照孔)×100％Inhibition rate (%) = [(OD positive control well- OD negative control well ) - (OD administration well - OD negative control well )] / (OD positive control well - OD negative control well ) × 100%
并据此按Logit法计算达到50％抑制率时的药物浓度，即IC50值。分子水平PARP2酶活性抑制实验According to this, the drug concentration at the 50% inhibition rate, that is, the IC 50 value, was calculated according to the Logit method. Molecular level PARP2 enzyme activity inhibition experiment
实验过程同分子水平PARP1酶活性抑制实验，所加酶和底物详细数据为：加入NAD+(终浓度80μM)，DNA(100ng/孔)，PARP2(10ng/孔)。The experimental procedure was the same as the inhibition of the PARP1 enzyme activity at the molecular level. The detailed data of the added enzyme and substrate were: addition of NAD + (final concentration 80 μM), DNA (100 ng/well), and PARP 2 (10 ng/well).
体外细胞增殖抑制实验In vitro cell proliferation inhibition assay
细胞株：中国仓鼠肺成纤维细胞株VC8(BRCA2-/-)Wiegant,W.W.；Overmeer,R.M.；Godthelp,B.C.；van Buul,P.and Zdzienicka,M.Z.Chinese hamster cell mutant,V-C8,a model for analysis of Brca2function.Mutation research 2006,600,79-88.和野生型细胞株V79(由荷兰莱顿大学Malgorzata Z.Zdzienicka教授赠送)。Cell line: Chinese hamster lung fibroblast cell line VC8 (BRCA2 -/- ) Wiegant, WW; Overmeer, RM; Godthelp, BC; van Buul, P. and Zdzienicka, MZ Chinese hamster cell mutant, V-C8, a model for analysis Of Brca2function.Mutation research 2006, 600, 79-88. and wild-type cell line V79 (given by Professor Malgorzata Z. Zdzienicka, Leiden University, The Netherlands).
实验方法：CCK8(Cell Counting Kit-8)法。CCK-8试剂中含有WST–8：化学名：2-(2-甲氧基-4-硝基苯基)-3-(4-硝基苯基)-5-(2,4-二磺酸苯)-2H-四唑单钠盐，它在电子载体1-甲氧基-5-甲基吩嗪硫酸二甲酯(1-Methoxy PMS)的作用下被细胞线粒体中的脱氢酶还原为具有高度水溶性的黄色甲臜产物(Formazan)。生成的甲臜物的数量与活细胞的数量成正比。细胞增殖越多越快，则颜色越深；细胞毒性越大，则颜色越浅。对于同样的细胞，颜色的深浅和细胞数目呈线性关系。用酶标仪在450nm波长处测定其光吸收值，可间接反映活细胞数量，从而计算出细胞抑制率。Experimental method: CCK8 (Cell Counting Kit-8) method. CCK-8 reagent contains WST-8: chemical name: 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfonate Acid benzene)-2H-tetrazole monosodium salt, which is reduced by dehydrogenase in cell mitochondria under the action of electron carrier 1-methoxy-5-methylphenazine dimethyl sulfate (1-Methoxy PMS) It is a yellow formazan product (Formazan) with high water solubility. The amount of formazan produced is proportional to the number of living cells. The more the cells proliferate, the darker the color; the greater the cytotoxicity, the lighter the color. For the same cells, the color depth and the number of cells are linear. The light absorption value was measured by a microplate reader at a wavelength of 450 nm, which indirectly reflected the number of living cells, thereby calculating the cell inhibition rate.
具体方法如下：将处于对数生长期的细胞按V79细胞2000个/孔，VC8细胞4000个/孔的密度接种至96孔培养板，每孔90μL，培养过夜后，加入不同浓度(终浓度从10μM开始，5倍稀释6个梯度)的药物10μL作用72h，每个浓度设三复孔，并设相应浓度的生理盐水溶媒对照及无细胞调零孔。作用结束后，加入10μL CCK8试剂，37℃继续培养4小时，酶标仪测OD450值。The specific method is as follows: the cells in the logarithmic growth phase are inoculated into the 96-well culture plate at a density of 2000 cells/well of V79 cells and 4000 cells/well of VC8 cells, 90 μL per well, and cultured overnight, and then added at different concentrations (final concentration from 10 μL of the drug was started at 10 μM, and 5 μL of 6 gradients were applied for 72 h. Three wells were set for each concentration, and the corresponding concentrations of physiological saline vehicle control and cell-free zero-adjusted wells were set. After the end of the action, 10 μL of CCK8 reagent was added, and the culture was continued at 37 ° C for 4 hours, and the OD450 value was measured by a microplate reader.
按照以下公式计算药物对细胞增殖抑制程度：Calculate the degree of inhibition of cell proliferation by the following formula:
抑制率(％)＝(OD对照孔-OD给药孔)/OD对照孔×100％Inhibition rate (%) = (OD control well- OD administration well ) / OD control well × 100%
并据此按Logit法计算达到50％抑制率时的药物浓度，即IC50值。According to this, the drug concentration at the 50% inhibition rate, that is, the IC 50 value, was calculated according to the Logit method.
实验结果表明：表1中发明的如下6个化合物对PARP1的抑制活性与阳性对照AZD2281相当，比阳性对照ABT-888活性要好，分别是I-4(14.04nM)，I-15(16.87nM)，I-22(14.33nM)，I-24(15.06nM)，I-28(7.52nM)，I-30(18.47nM)，I-32(19.68nM)。而且大部分化合物对PARP1表现出一定的选择性，对PARP1抑制作用强于PARP2，不同于AZD-2281与ABT-888。表2中化合物有3个化合物对PARP1的抑制作用于阳性化合物AZD2281相当，IC50(nM)分别为:I-37(18.03nM)，I-39(33.50nM)，I-44(17.78nM)；对PARP2的作用强度均弱于阳性对照AZD2281，但部分化合物对PARP2的作用强度强于PARP1，显示对PARP2的选择性。体外增殖实验表明：表1和2中化合物对中国仓鼠肺成纤维细胞株VC8(BRCA2-/-)具有良好的抑制活性，而 对野生型无缺陷的中国仓鼠肺细胞株VC79(BRCA2+/+)无抑制活性，表明所设计化合物对BRCA缺陷的细胞具有高度的选择性。The experimental results showed that the following six compounds invented in Table 1 had the same inhibitory activity against PARP1 as the positive control AZD2281, which was better than the positive control ABT-888, which was I-4 (14.04 nM) and I-15 (16.87 nM). I-22 (14.33 nM), I-24 (15.06 nM), I-28 (7.52 nM), I-30 (18.47 nM), I-32 (19.68 nM). Moreover, most of the compounds showed some selectivity for PARP1, and PARP1 had stronger inhibition than PARP2, unlike AZD-2281 and ABT-888. The inhibitory effect of 3 compounds on PARP1 in Table 2 was comparable to that of the positive compound AZD2281, IC 50 (nM): I-37 (18.03 nM), I-39 (33.50 nM), I-44 (17.78 nM). The intensity of action on PARP2 was weaker than that of the positive control AZD2281, but some compounds exerted stronger effects on PARP2 than PARP1, indicating selectivity for PARP2. In vitro proliferation experiments showed that the compounds in Tables 1 and 2 had good inhibitory activity against Chinese hamster lung fibroblast cell line VC8 (BRCA2 -/- ), whereas the wild-type non-defective Chinese hamster lung cell line VC79 (BRCA2 +/+) There is no inhibitory activity, indicating that the designed compound is highly selective for BRCA-deficient cells.
表1.部分苯并咪唑类化合物的分子水平和细胞水平生物活性结果Table 1. Molecular and cellular levels of biological activity of some benzimidazoles
Figure PCTCN2016087368-appb-000217
Figure PCTCN2016087368-appb-000217
表2.化合物I-36～I-45的分子水平和细胞水平生物活性结果Table 2. Molecular and Cellular Bioactivity Results of Compounds I-36 to I-45
Figure PCTCN2016087368-appb-000218
Figure PCTCN2016087368-appb-000218
Figure PCTCN2016087368-appb-000219
Figure PCTCN2016087368-appb-000219
注：表中“--”表示最高浓度无效或无计算结果。Note: “--” in the table indicates that the highest concentration is invalid or there is no calculation result.
从表1中可知，当用氧原子代替硫原子时(化合物I-34对化合物I-15)，由于分子内不存在硫原子与苯并咪唑氮原子之间的n0—σ*非键作用，导致化合物I-34对PARP1的活性急剧下降。It can be seen from Table 1 that when an oxygen atom is used in place of a sulfur atom (compound I-34 to compound I-15), n 0 —σ* non-bonding between a sulfur atom and a benzimidazole nitrogen atom is absent in the molecule. This results in a sharp decrease in the activity of compound I-34 against PARP1.
化合物I-35与I-15相比，采用了开环策略，其对PARP1的抑制活性也有所下降，但对PARP2的活性则有所增强。Compared with I-15, compound I-35 adopts a ring-opening strategy, and its inhibitory activity against PARP1 is also decreased, but the activity of PARP2 is enhanced.
化合物I-30单盐酸盐对人乳腺癌MDA-MB-436裸小鼠皮下移植瘤的生长抑制作用如下：The inhibitory effect of Compound I-30 monohydrochloride on the growth of human breast cancer MDA-MB-436 nude mice was as follows:
实验方法experimental method
动物：BALB/cA裸小鼠，雌性，4-5周龄，体重19±2g，由中国科学院上海药物研究所提供，生产许可证编号：SCXK(沪)2013-001。使用合格证编号：SYXK(沪)2013-0049。每组动物数：阴性对照组12只，给药组6只。细胞株：人乳腺癌MDA-MB-436细胞株接种至裸小鼠右侧腋窝皮下，细胞接种量为5×106/只，形成移植瘤后再在裸小鼠体内传2代后使用。Animals: BALB/cA nude mice, female, 4-5 weeks old, weighing 19±2 g, provided by Shanghai Institute of Materia Medica, Chinese Academy of Sciences, production license number: SCXK (Shanghai) 2013-001. Use certificate number: SYXK (Shanghai) 2013-0049. The number of animals in each group: 12 in the negative control group and 6 in the drug-administered group. Cell line: Human breast cancer MDA-MB-436 cell line was inoculated subcutaneously into the right axilla of nude mice, and the amount of cells inoculated was 5×10 6 /piece. After the transplanted tumor was formed, it was used in nude mice for 2 generations.
取生长旺盛期的瘤组织剪切成1.5mm3左右，在无菌条件下，接种于裸小鼠右侧腋窝皮下。裸小鼠皮下移植瘤用游标卡尺测量移植瘤直径，待肿瘤平均体积生长至200mm3左右后，将动物随机分组。I-30单盐酸盐100mg/kg和20mg/kg组，每天口服给药一次，连续给药21天。阳性对照药物AZD2281 30mg/kg，每天口服给药一次，连续给药21天。溶剂对照给等量注射用水。The tumor tissue in the vigorous growth period was cut into 1.5 mm 3 and inoculated subcutaneously in the right axilla of nude mice under aseptic conditions. The diameter of the transplanted tumor was measured with a vernier caliper in nude mice. After the average tumor volume grew to about 200 mm 3 , the animals were randomly divided into groups. The I-30 monohydrochloride 100 mg/kg and 20 mg/kg groups were orally administered once a day for 21 consecutive days. The positive control drug AZD2281 30 mg/kg was orally administered once a day for 21 days. The solvent control was given an equal amount of water for injection.
整个实验过程中，每周2次测量移植瘤直径，同时称量小鼠体重。The diameter of the transplanted tumor was measured twice a week during the entire experiment, and the body weight of the mice was weighed.
肿瘤体积(tumor volume,TV)的计算公式为：The formula for calculating tumor volume (TV) is:
TV＝1/2×a×b2 TV = 1/2 × a × b 2
其中a、b分别表示长、宽。Where a and b represent length and width, respectively.
根据测量的结果计算出相对肿瘤体积(relative tumor volume，RTV)，计算公式为：Calculate the relative tumor volume (RTV) based on the measured results, and the formula is:
RTV＝Vt/V0 RTV=V t /V 0
其中V0为分笼给药时(即d0)测量所得肿瘤体积，Vt为每一次测量时的肿瘤体积。Where V 0 is the measured tumor volume at the time of sub-cage administration (i.e., d 0 ), and V t is the tumor volume at each measurement.
抗肿瘤活性的评价指标为：The evaluation index of antitumor activity is:
1)相对肿瘤增殖率T/C(％)，计算公式如下：1) Relative tumor growth rate T/C (%), the formula is as follows:
T/C(％)＝(TRTV/CRTV)×100％T/C(%)=(T RTV /C RTV )×100%
TRTV：治疗组RTV；CRTV：阴性对照组RTV；T RTV : treatment group RTV; C RTV : negative control group RTV;
2)肿瘤体积增长抑制率GI％，计算公式如下：2) Tumor volume growth inhibition rate GI%, calculated as follows:
GI％＝[1-(TVt-TV0)/(CVt-CV0)]×100％GI%=[1-(TVt-TV 0 )/(CVt-CV 0 )]×100%
TVt为治疗组每次测量的瘤体积；TV0为治疗组分笼给药时所得瘤体积；CVt为对照组每次测量的瘤体积；CV0为对照组分笼给药时所得瘤体积； TVt is the tumor volume measured in each treatment group; TV 0 is the tumor volume obtained when the therapeutic component is administered in cage; CVt is the tumor volume measured in each control group; CV 0 is the tumor volume obtained when the control group is administered in cages;
3)瘤重抑制率，计算公式如下：3) The tumor weight inhibition rate is calculated as follows:
瘤重抑制率％＝(Wc-WT)/Wc×100％Tumor weight inhibition rate%=(Wc-W T )/Wc×100%
Wc：对照组瘤重，WT：治疗组瘤重。Wc: The control group was heavier, W T : the tumor weight of the treatment group.
结果如表3所示，I-30单盐酸盐每天口服给药100mg/kg或20mg/kg，一周后肿瘤生长明显减缓，连续给药三周后能显著抑制人乳腺癌MDA-MB-436裸小鼠移植瘤的生长，第21天的T/C为2.12％和26.24％，其抑制肿瘤生长的效果优于AZD2281 30mg/kg组。表2显示，I-30单盐酸盐每天口服给药100mg/kg或20mg/kg对裸小鼠的体重影响较小。The results are shown in Table 3. I-30 monohydrochloride was orally administered 100 mg/kg or 20 mg/kg per day. Tumor growth was significantly slowed after one week. After three weeks of continuous administration, human breast cancer MDA-MB-436 was significantly inhibited. The growth of nude mice xenografts was 2.12% and 26.24% on day 21, and its effect on inhibiting tumor growth was better than that of AZD2281 30 mg/kg group. Table 2 shows that oral administration of 100 mg/kg or 20 mg/kg per day of I-30 monohydrochloride has less effect on the body weight of nude mice.
表3. I-30单盐酸盐对人乳腺癌MDA-MB-436裸小鼠移植瘤的相对肿瘤增殖率Table 3. Relative tumor growth rate of I-30 monohydrochloride for human breast cancer MDA-MB-436 nude mice xenografts
Figure PCTCN2016087368-appb-000220
Figure PCTCN2016087368-appb-000220
(*p<0.05 **p<0.001)(*p<0.05 **p<0.001)
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。 All documents mentioned in the present application are hereby incorporated by reference in their entirety in their entireties in the the the the the the the the In addition, it should be understood that various modifications and changes may be made by those skilled in the art in the form of the appended claims.

Claims (12)

  1. 一种如通式Ia所示的化合物、R-异构体、S-异构体、或其药学上可接受的盐：A compound of the formula Ia, an R-isomer, an S-isomer, or a pharmaceutically acceptable salt thereof:
    Figure PCTCN2016087368-appb-100001
    Figure PCTCN2016087368-appb-100001
    其中，among them,
    R为氢、C1-C4的直链或支链烷基；R is hydrogen, a linear or branched alkyl group of C1-C4;
    R1为氢、取代或未取代的C1-C4的直链或支链烷基、或取代或未取代的C3-C6的环烷基；R 1 is hydrogen, a substituted or unsubstituted C1-C4 linear or branched alkyl group, or a substituted or unsubstituted C3-C6 cycloalkyl group;
    R2为无、卤素、取代或未取代的C1-C4的直链或支链烷基、或取代或未取代的C3-C6的环烷基；R 2 is a free, halogen, substituted or unsubstituted C1-C4 linear or branched alkyl group, or a substituted or unsubstituted C3-C6 cycloalkyl group;
    Y1、Y2独立地选自取代或未取代的亚甲基、取代或未取代的亚乙基、取代或未取代的亚丙基或取代或未取代的亚丁基；Y 1 , Y 2 are independently selected from substituted or unsubstituted methylene, substituted or unsubstituted ethylene, substituted or unsubstituted propylene or substituted or unsubstituted butylene;
    R3为氢、取代或未取代的C1-C4的直链或支链烷基、-C(＝O)R4、-SO2R5或取代或未取代的C3-C6的环烷基，其中，R4、R5独立地为取代或未取代的C1-C4的直链或支链烷基、取代或未取代的C3-C6的环烷基或C6-C10芳基；R 3 is hydrogen, a substituted or unsubstituted C1-C4 linear or branched alkyl group, -C(=O)R 4 , -SO 2 R 5 or a substituted or unsubstituted C3-C6 cycloalkyl group, Wherein R 4 and R 5 are independently a substituted or unsubstituted C1-C4 linear or branched alkyl group, a substituted or unsubstituted C3-C6 cycloalkyl group or a C6-C10 aryl group;
    X为CR6或N，其中R6为氢、取代或未取代的C1-C4的直链或支链烷基、取代或未取代的C1-C4的直链或支链烷氧基、卤素或取代或未取代的C3-C6的环烷基；X is CR 6 or N, wherein R 6 is hydrogen, a substituted or unsubstituted C1-C4 linear or branched alkyl group, a substituted or unsubstituted C1-C4 linear or branched alkoxy group, halogen or a substituted or unsubstituted C3-C6 cycloalkyl group;
    其中所述各取代独立地指具有选自下组的1-3个取代基：羟基、卤素、C1-C6直链或支链烷基、C1-C4直链或支链烷氧基、C3-C6的环烷基、C6-C10芳基、羧基；所述取代基中的C1-C6直链或支链烷基可进一步被氨基、羟基、甲氧基、乙氧基、丙氧基、异丙氧基、-CO2Me、-CO2Et、-CO2Pr或-CO2Pr-i取代。Wherein each substitution independently refers to having from 1 to 3 substituents selected from the group consisting of hydroxy, halo, C1-C6 straight or branched alkyl, C1-C4 straight or branched alkoxy, C3- a cycloalkyl group, a C6-C10 aryl group or a carboxyl group of C6; the C1-C6 linear or branched alkyl group in the substituent may be further amino, hydroxy, methoxy, ethoxy, propoxy or iso Propyl, -CO 2 Me, -CO 2 Et, -CO 2 Pr or -CO 2 Pr-i are substituted.
  2. 如权利要求1所述的化合物、R-异构体、S-异构体、或其药学上可接受的盐，其特征在于，所述化合物如通式I所示：The compound, R-isomer, S-isomer, or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is as shown in Formula I:
    Figure PCTCN2016087368-appb-100002
    Figure PCTCN2016087368-appb-100002
    其中，among them,
    R1为氢、取代或未取代的C1-C4的直链或支链烷基、或取代或未取代的C3-C6的环烷基；R 1 is hydrogen, a substituted or unsubstituted C1-C4 linear or branched alkyl group, or a substituted or unsubstituted C3-C6 cycloalkyl group;
    R2为无、卤素、取代或未取代的C1-C4的直链或支链烷基、或取代或未取代的C3-C6的环烷基；R 2 is a free, halogen, substituted or unsubstituted C1-C4 linear or branched alkyl group, or a substituted or unsubstituted C3-C6 cycloalkyl group;
    Y1、Y2独立地选自取代或未取代的亚甲基、取代或未取代的亚乙基、取代或未取 代的亚丙基或取代或未取代的亚丁基；Y 1 , Y 2 are independently selected from substituted or unsubstituted methylene, substituted or unsubstituted ethylene, substituted or unsubstituted propylene or substituted or unsubstituted butylene;
    R3为氢、取代或未取代的C1-C4的直链或支链烷基、-C(＝O)R4、-SO2R5或取代或未取代的C3-C6的环烷基，其中，R4、R5独立地为取代或未取代的C1-C4的直链或支链烷基、取代或未取代的C3-C6的环烷基或C6-C10芳基；R 3 is hydrogen, a substituted or unsubstituted C1-C4 linear or branched alkyl group, -C(=O)R 4 , -SO 2 R 5 or a substituted or unsubstituted C3-C6 cycloalkyl group, Wherein R 4 and R 5 are independently a substituted or unsubstituted C1-C4 linear or branched alkyl group, a substituted or unsubstituted C3-C6 cycloalkyl group or a C6-C10 aryl group;
    X为CR6或N，其中R6为氢、取代或未取代的C1-C4的直链或支链烷基或取代或未取代的C3-C6的环烷基；X is CR 6 or N, wherein R 6 is hydrogen, a substituted or unsubstituted C1-C4 linear or branched alkyl group or a substituted or unsubstituted C3-C6 cycloalkyl group;
    其中所述各取代独立地指具有选自下组的1-3个取代基：羟基、卤素、C1-C6直链或支链烷基、C1-C4直链或支链烷氧基、C3-C6的环烷基、C6-C10芳基、羧基。Wherein each substitution independently refers to having from 1 to 3 substituents selected from the group consisting of hydroxy, halo, C1-C6 straight or branched alkyl, C1-C4 straight or branched alkoxy, C3- a cycloalkyl group of C6, a C6-C10 aryl group, or a carboxyl group.
  3. 如权利要求2所述的化合物，其特征在于，R1为氢、C1-C4的直链或支链烷基、卤素取代的C1-C4的直链或支链烷基或C3-C6的环烷基。The compound according to claim 2, wherein R 1 is hydrogen, a C1-C4 linear or branched alkyl group, a halogen-substituted C1-C4 linear or branched alkyl group or a C3-C6 ring. alkyl.
  4. 如权利要求2所述的化合物，其特征在于，R2为无、氟、氯、溴、C1-C4的直链或支链烷基、或C3-C6的环烷基。The compound according to claim 2, wherein R 2 is a free or fluorine, chlorine, bromine, C1-C4 linear or branched alkyl group, or a C3-C6 cycloalkyl group.
  5. 如权利要求2所述的化合物，其特征在于，R4、R5独立地为取代或未取代的C1-C4的直链或支链烷基、C3-C6的环烷基或苯基，所述取代是指具有选自下组的1-2个取代基：羟基、卤素、C1-C4直链或支链烷氧基。The compound according to claim 2, wherein R 4 and R 5 are independently a substituted or unsubstituted C1-C4 linear or branched alkyl group, a C3-C6 cycloalkyl group or a phenyl group. By substitution is meant having one or two substituents selected from the group consisting of hydroxy, halo, C1-C4 straight or branched alkoxy.
  6. 如权利要求2所述的化合物，其特征在于，R6为氢、C1-C4的直链或支链烷基或C3-C6的环烷基。The compound according to claim 2, wherein R 6 is hydrogen, a C1-C4 linear or branched alkyl group or a C3-C6 cycloalkyl group.
  7. 如权利要求2所述的化合物，其特征在于，Y1为取代或未取代的亚甲基、Y2取代或未取代的亚乙基；或者Y1为取代或未取代的亚乙基、Y2为取代或未取代的亚甲基，其中The compound according to claim 2, wherein Y 1 is a substituted or unsubstituted methylene group, a Y 2 -substituted or unsubstituted ethylene group; or Y 1 is a substituted or unsubstituted ethylene group, Y 2 is a substituted or unsubstituted methylene group, wherein
    所述各取代独立地指具有选自下组的1-2个取代基：羟基、卤素、C1-C4直链或支链烷基、C1-C4直链或支链烷氧基、C3-C6的环烷基；其中的C1-C4直链或支链烷基可进一步被氨基、羟基、甲氧基、乙氧基、丙氧基、异丙氧基、-CO2Me、-CO2Et、-CO2Pr或-CO2Pr-i取代。Each of the substituents independently means having 1-2 substituents selected from the group consisting of hydroxy, halogen, C1-C4 straight or branched alkyl, C1-C4 straight or branched alkoxy, C3-C6 a cycloalkyl group; wherein the C1-C4 straight or branched alkyl group may be further amino, hydroxy, methoxy, ethoxy, propoxy, isopropoxy, -CO 2 Me, -CO 2 Et , -CO 2 Pr or -CO 2 Pr-i substituted.
  8. 如权利要求1所述的化合物，其特征在于，所述化合物选自下组： The compound of claim 1 wherein said compound is selected from the group consisting of:
    Figure PCTCN2016087368-appb-100003
    Figure PCTCN2016087368-appb-100003
    Figure PCTCN2016087368-appb-100004
    Figure PCTCN2016087368-appb-100004
  9. 一种如通式(IIa)所示的化合物，a compound of the formula (IIa),
    Figure PCTCN2016087368-appb-100005
    Figure PCTCN2016087368-appb-100005
    其中，R1、R2、Y1、Y2、R3、X的定义如权利要求1所述，R7为C1-C4的直链或支链烷基。Wherein R 1 , R 2 , Y 1 , Y 2 , R 3 , X are as defined in claim 1, and R 7 is a C1-C4 linear or branched alkyl group.
  10. 如权利要求9所述的化合物，其特征在于，所述化合物如式II所示：The compound of claim 9 wherein said compound is as shown in formula II:
    Figure PCTCN2016087368-appb-100006
    Figure PCTCN2016087368-appb-100006
  11. 一种药物组合物，其特征在于，所述药物组合物包括权利要求1所述的化合物、R-异构体、S-异构体、或其药学上可接受的盐；以及A pharmaceutical composition comprising the compound according to claim 1, the R-isomer, the S-isomer, or a pharmaceutically acceptable salt thereof;
    药学上可接受的载体。A pharmaceutically acceptable carrier.
  12. 如权利要求1所述的化合物或如权利要求11所述的药物组合物的用途，其特征在于，用于制备：(1)PARP1抑制剂；(2)PARP2抑制剂；(3)预防和/或治疗肿瘤的药物；(4)抗炎药物；和/或(5)预防和/或治疗与PARP相关疾病的药物。 Use of a compound according to claim 1 or a pharmaceutical composition according to claim 11 for the preparation of: (1) a PARP1 inhibitor; (2) a PARP2 inhibitor; (3) prevention and/or Or a drug for treating a tumor; (4) an anti-inflammatory drug; and/or (5) a drug for preventing and/or treating a disease associated with PARP.
PCT/CN2016/087368 2015-06-26 2016-06-27 2-substituted benzimidazole-4-carboxamide compound and preparation method and use thereof WO2016206651A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201680037353.6A CN107922430B (en) 2015-06-26 2016-06-27 2-substituted benzimidazole-4-formamide compound and preparation method and application thereof

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201510363842.8A CN106317069A (en) 2015-06-26 2015-06-26 2-substituted-benzimidazole-4-formamide compound, preparation method, and application thereof
CN201510363842.8 2015-06-26

Publications (1)

Publication Number Publication Date
WO2016206651A1 true WO2016206651A1 (en) 2016-12-29

Family

ID=57584689

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2016/087368 WO2016206651A1 (en) 2015-06-26 2016-06-27 2-substituted benzimidazole-4-carboxamide compound and preparation method and use thereof

Country Status (2)

Country Link
CN (2) CN106317069A (en)
WO (1) WO2016206651A1 (en)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001021615A1 (en) * 1999-09-17 2001-03-29 Yamanouchi Pharmaceutical Co., Ltd. Benzimidazole derivatives
CN101155797A (en) * 2005-04-11 2008-04-02 艾博特公司 1h-benzimidazole-4-carboxamides substituted with a quaternary carbon at the 2-position are potent parp inhibitors

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
HU0301154D0 (en) * 2003-04-28 2003-07-28 Hideg Kalman Dr Pharmaceutical composition

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001021615A1 (en) * 1999-09-17 2001-03-29 Yamanouchi Pharmaceutical Co., Ltd. Benzimidazole derivatives
CN101155797A (en) * 2005-04-11 2008-04-02 艾博特公司 1h-benzimidazole-4-carboxamides substituted with a quaternary carbon at the 2-position are potent parp inhibitors

Also Published As

Publication number Publication date
CN107922430B (en) 2020-09-04
CN106317069A (en) 2017-01-11
CN107922430A (en) 2018-04-17

Similar Documents

Publication Publication Date Title
TWI697486B (en) 3-aryl-4-amido-bicyclic [4,5,0] hydroxamic acids as hdac inhibitors
TW201014855A (en) Compounds for the treatment of hepatitis C
WO2020259432A1 (en) Kras-g12c inhibitor
EA014956B1 (en) Heterocyclic compounds useful for the treatment of inflammatory and allergic disorders
WO2019020070A1 (en) Piperazine heteroaryl derivative, preparation method therefor and use of same in medicine
WO2001083456A1 (en) Condensed heteroaryl derivatives
WO2023274383A1 (en) Kras g12d inhibitor and use thereof
JP2021519308A (en) Imidazopiperazin inhibitor of transcriptional activation protein
TW201636329A (en) Bicyclic [4,6,0] hydroxamic acids as HDAC inhibitors
JP2019065020A (en) Kynurenine-3-monooxygenase inhibitor, pharmaceutical composition and application method therefor
WO2019161803A1 (en) Peptidylarginine deiminase inhibitor and use thereof
WO2023098832A1 (en) Pyridopyrimidine derivatives serving as small gtp enzyme kras mutation inhibitors
JP2016504299A (en) Use of maleimide derivatives to prevent and treat leukemia
WO2013107326A1 (en) NEW-TYPE IMIDAZO [1, 2-a] PYRIDINE COMPOUNDS, PREPARATION METHOD THEREFOR, PHARMACEUTICAL COMPOSITION CONTAINING SAME AND USE THEREOF
AU2021208915B2 (en) Novel pyrazole derivative
CN103298791B (en) Viral polymerase inhibitors
TWI804295B (en) Compounds as methionine adenosyltransferase inhibitors, preparation methods and applications thereof
CN107922430B (en) 2-substituted benzimidazole-4-formamide compound and preparation method and application thereof
CN111171041B (en) 20-substituted camptothecin derivative and preparation method and application thereof
WO2022171198A1 (en) Influenza virus inhibitor and use thereof
TW202208374A (en) Substituted tricyclic amides, analogues thereof, and methods using same
WO2015114660A1 (en) Novel heterobicyclic compounds as kappa opioid agonists
US20190135828A1 (en) Hepatitis c virus inhibitor and uses thereof
WO2023246837A1 (en) Class of compounds having pyrimido-six-membered ring structure, pharmaceutical compositions comprising same, and use thereof
TW201109335A (en) Heterocyclic compounds

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 16813767

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 16813767

Country of ref document: EP

Kind code of ref document: A1