WO2021121396A1 - Tricyclic compound that acts as plasma kallikrein inhibitor and use thereof - Google Patents

Tricyclic compound that acts as plasma kallikrein inhibitor and use thereof Download PDF

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WO2021121396A1
WO2021121396A1 PCT/CN2020/137640 CN2020137640W WO2021121396A1 WO 2021121396 A1 WO2021121396 A1 WO 2021121396A1 CN 2020137640 W CN2020137640 W CN 2020137640W WO 2021121396 A1 WO2021121396 A1 WO 2021121396A1
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methyl
compound
benzo
solvate
dihydro
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Chinese (zh)
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强晓明
万剑飞
刘婷
吴红丽
柯潇
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成都康弘药业集团股份有限公司
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    • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • the present invention relates to tricyclic compounds with selective inhibitory activity on plasma kallikrein and uses thereof.
  • Plasma kallikrein is a member of the serine protease family and was first discovered in the plasma of mammals. It is encoded by a single gene (KLKB1) located on the 4q35 chromosome. It is mainly synthesized in the liver and is abundantly present in the blood circulation in the form of plasma kallikrein (PPK). PPK further cleaves its intrinsic Arg- by coagulation factor XIIa. IIe key activation is transformed into PK (Yousef, GMet al. An overview of the kallikrein gene families in humans and other species: Emerging candidates tumor markers. Clin. Biochem. 2003, 36, 443-452).
  • PK is a key enzyme of the kallikrein-kinin system (KKS), which can act on high molecular weight kininogen (KH) to activate the release of small-molecule bradykinin (BK), and then act on bradykinin Peptide receptors are involved in biological processes such as blood coagulation, fibrinolysis, complement activation and inflammation.
  • KS kallikrein-kinin system
  • hereditary angioedema which is an autosomal dominant inheritance
  • HAE hereditary angioedema
  • Icaratide and Lanadelumab have been marketed clinically for the treatment of hereditary angioedema, with significant curative effects.
  • Biocryst developed the oral plasma kallikrein inhibitor BCX4161 (Collis, P.et al., BCX4161, an oral kallikrein inhibitor: safety and pharmacokinetic results of a Phase 1 study in healthy volunteers.J.Allergy Clin.Immunol 2014, 133, 133, Issue 2, Supplement, 2014, AB39), but its clinical administration dose is 400 mg each time, three times a day.
  • the benzamidine or guanidine group constitutes the pharmacodynamic group of BCX4161 in the structure, its poor physical and chemical properties may affect the bioavailability of the drug, so it needs to be administered in a larger dose.
  • the existing plasma kallikrein inhibitors also have the problem of poor selectivity to related enzymes such as KLK1, thrombin and other serine proteases. So far, no small molecule plasma kallikrein inhibitor has been approved for marketing. Therefore, there is still a need to study new types of selective plasma kallikrein inhibitors with stronger effects and fewer side effects.
  • the present invention relates to a plasma kallikrein inhibitor compound, a pharmaceutical composition of the compound, the pharmaceutical use of the compound, and a treatment method of the compound.
  • the present invention provides a compound of formula (I), a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof:
  • A is selected from a 5-membered or 6-membered aromatic ring or an aromatic heterocyclic ring, the aromatic heterocyclic ring containing 1-3 heteroatoms selected from N, O and S, and the aromatic ring or aromatic heterocyclic ring is optionally Substituent substitution: halogen, alkyl, alkoxy, haloalkyl, OH, CN, COOR 1 , CONR 1 R 2 , NR 1 R 2 , NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1-3 OR 1 and C(R 1 )(R 2 )NH 2 ;
  • Or A is selected from a fused 6,5- or 6,6-aromatic heterobicyclic ring containing N and optionally additionally independently 1-2 heteroatoms selected from N, O and S, so
  • the heterobicyclic ring is optionally substituted by the following substituents: halogen, alkyl, alkoxy, haloalkyl, OH, CN, COOR 1 , CONR 1 R 2 , NR 1 R 2 , NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1-3 OR 1 and -C(R 1 )(R 2 )NH 2 ;
  • L 1 is selected from (CR 3 R 4 ) m or (CR 3 R 4 ) m O, where m is 0, 1, 2;
  • X 1 and X 2 are CR 5 or N, and are not N at the same time; wherein R 5 is selected from H, OH, halogen, alkyl, alkoxy, haloalkyl, cycloalkyl, CN, COOR 1 , CONR 1 R 2.
  • L 2 is selected from bond, (CH 2 ) n , (CH 2 ) n NH, (CH 2 ) n O, where n is 1 or 2;
  • B is selected from a 5-6 membered aromatic ring or an aromatic heterocyclic ring, wherein the aromatic heterocyclic ring contains 1-3 heteroatoms selected from N, O or S.
  • the aromatic ring or aromatic heterocyclic ring is alkylated by an alkyl group. , Alkoxy, OH, halogen, CN, ketone, COOR 1 , CONR 1 R 2 , NR 1 R 2 , NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1- 3 OR 1 and -C(R 1 )(R 2 )NH 2 are substituted;
  • Or B is selected from a saturated or unsaturated 5-10 membered monocyclic or fused polycyclic non-aromatic ring system, the non-aromatic ring system optionally contains 0-3 selected from N, O and S Heteroatoms, the non-aromatic ring system is optionally substituted by the following substituents: halogen, alkyl, alkoxy, haloalkyl, keto, OH, CN, COOR 1 , CONR 1 R 2 , NR 1 R 2. NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1-3 OR 1 and -C(R 1 )(R 2 )NH 2 ;
  • R 1 and R 2 are independently selected from H and alkyl, or R 1 and R 2 together with the carbon to which they are attached form a cycloalkyl;
  • R 3 and R 4 are independently selected from H and alkyl, or R 3 and R 4 together with the carbon to which they are attached form a cycloalkyl.
  • A is selected from wherein, X 3 is CR 10 or N, R 6 , R 7 , R 8 , R 9 , R 10 are independently selected from hydrogen, halogen, alkyl, alkoxy, haloalkyl, OH, CN, COOR 1 , CONR 1 R 2 , NR 1 R 2 , NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1-3 OR 1 and C(R 1 )(R 2 )NH 2 ; R 1 And R 2 are independently selected from H and an alkyl group, or R 1 and R 2 together with the carbon to which they are attached form a cycloalkyl group.
  • R 8 is NH 2 or C(R 1 )(R 2 )NH 2 , wherein R 1 and R 2 are independently selected from H and C 1-3
  • the alkyl group, or R 1 and R 2 together with the carbon to which they are attached form a 3-membered or 4-membered cycloalkyl group.
  • R 8 is NH 2 .
  • R 8 is CH 2 NH 2 .
  • R 8 is C(R 1 )(R 2 )NH 2 , wherein R 1 and R 2 together with the carbon to which they are attached form a cyclopropyl group.
  • R 6 , R 7 , R 9 , R 10 are independently selected from H, halogen, alkyl, alkoxy, haloalkyl.
  • R 6 , R 7 , R 9 , R 10 are independently selected from H, halogen, or CH 3 .
  • A is selected from:
  • A is selected from isoquinoline and 1H-pyrrole [2,3-b]pyridine, wherein isoquinoline and 1H-pyrrole [2,3-b] Pyridine is optionally substituted by halogen, alkyl, alkoxy, haloalkyl, OH, CN, COOR 1 , CONR 1 R 2 , NR 1 R 2 , NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1-3 OR 1 and C(R 1 )(R 2 )NH 2 substitution; R 1 and R 2 are independently selected from H and alkyl, or R 1 and R 2 and the carbon to which they are attached Together to form a cycloalkyl group.
  • A is selected from isoquinoline and 1H-pyrrole [2,3-b]pyridine, wherein isoquinoline and 1H-pyrrole [2,3-b] Pyridine is optionally substituted with halogen, alkyl, alkoxy, haloalkyl, and amino.
  • A is selected from isoquinoline and 1H-pyrrole [2,3-b]pyridine, wherein isoquinoline and 1H-pyrrole [2,3-b] Pyridine is optionally substituted with halogen, alkyl, alkoxy, haloalkyl, and amino.
  • A is selected from:
  • L 1 is selected from bond, CH 2 , (CH 2 ) 2 O.
  • L 1 is selected from CH 2 .
  • X 2 is CR 5 , wherein R 5 is selected from H, alkyl, haloalkyl, and alkoxy.
  • X 2 is CR 5 , wherein R 5 is selected from H, CH 3 , CF 3 , CH 2 OCH 3 .
  • L 2 is selected from bond, O, NH, CH 2 , CH 2 NH, CH 2 O.
  • L 2 is selected from CH 2 .
  • B is selected from:
  • the compound of formula (I) is selected from:
  • the second object of the present invention is to provide a pharmaceutical composition
  • a pharmaceutical composition comprising the compound according to the first object of the present invention, a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof, and a pharmaceutical composition. Acceptable excipients.
  • the third object of the present invention is to provide the compound according to the first object of the present invention, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug, which is used in the treatment or prevention of plasma kallikrein. Use in medicine for active diseases or conditions.
  • the fourth object of the present invention is to provide the compound according to the first object of the present invention, its solvate, tautomer or pharmaceutically acceptable salt for the treatment or prevention of diseases or diseases involving plasma kallikrein activity in mammals
  • the method of disease includes administering a therapeutically effective amount of the compound described in the first object of the present invention to a mammal in need thereof.
  • the disease involving plasma kallikrein activity is inflammation.
  • the disease involving plasma kallikrein activity is selected from the group consisting of impaired vision, diabetic retinopathy, diabetic macular edema, hereditary angioedema, diabetes, pancreatitis, cerebral hemorrhage, nephropathy, cardiomyopathy, neuropathy, Inflammatory bowel disease, arthritis, septic shock, hypotension, cancer, adult respiratory distress syndrome, diffuse intravascular coagulation, cardiopulmonary bypass surgery and bleeding after surgery.
  • the disease involving plasma kallikrein activity is a retinal vascular permeability disease associated with diabetic retinopathy and diabetic macular edema.
  • the disease involving plasma kallikrein activity is diabetic macular edema.
  • the disease involving plasma kallikrein activity is hereditary angioedema.
  • Co-administration therapy includes a compound of formula (I) and one or more other therapeutic agents, and the other therapeutic agents are selected from: therapeutic agents that inhibit platelet-derived factor (PDGF), vascular endothelial growth factor (VEGF), integrin alpha 5 ⁇ 1 , steroids, other therapeutic agents that inhibit plasma kallikrein, and other inflammation inhibitors.
  • PDGF platelet-derived factor
  • VEGF vascular endothelial growth factor
  • integrin alpha 5 ⁇ 1 a compound of formula (I) and one or more other therapeutic agents
  • steroids other therapeutic agents that inhibit plasma kallikrein, and other inflammation inhibitors.
  • the compound of the present invention and the combined therapeutic agent may be present in the same or different pharmaceutical compositions, and may be administered separately, sequentially or simultaneously.
  • the compound of the present invention can be administered in combination with laser treatment of the retina.
  • Laser photocoagulation combined with anti-VEGF drugs such as ranibizumab
  • ranibizumab in the treatment of diabetic macular edema is well known in the art and has been proven to have significant efficacy.
  • alkyl by itself or as part of another substituent means (unless otherwise stated) a saturated linear or branched hydrocarbon group having the specified number of carbon atoms. Including: up to 10 carbon atoms (C 1 -C 10 ), or up to 6 carbon atoms (C 1 -C 6 ), or up to 4 carbon atoms (C 1 -C 4 ), or up to 3 carbon atoms ( C 1 -C 3 ) linear group; or 3 to 10 carbon atoms (C 3 -C 10 ), or up to 7 carbon atoms (C 3 -C 7 ), or up to 4 carbon atoms (C 3 -C 4 ) branched chain group.
  • alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n- Sinki.
  • alkoxy represents the aforementioned alkyl group having a specified number of carbon atoms connected by an oxygen bridge. Including: 1 to 6 carbon atoms (C 1 -C 6 ), or 1 to 4 carbon atoms (C 1 -C 4 ) linear groups; -3 to 6 carbon atoms (C 3 -C 6 ) , Or a branched group with 3 to 4 carbon atoms (C 3 -C 4 ).
  • alkoxy groups include, but are not limited to: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy and S- Pentoxy.
  • cycloalkyl is a monocyclic saturated hydrocarbon group of 3 to 7 carbon atoms.
  • the cycloalkyl group may contain 3 to 7 carbon atoms or 3 to 6 carbon atoms or 3 to 5 carbon atoms or 3 to 4 carbon atoms.
  • monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.
  • haloalkyl is a group formed by partially or completely replacing hydrogen atoms on an alkyl group with halogen atoms as described above; unless otherwise specified, halogen is selected from Cl, F, Br, and I.
  • aromatic ring refers to a monocyclic, bicyclic or polycyclic aromatic hydrocarbon group, such as benzene, naphthalene.
  • aryl also includes two or more ring systems in which two or more carbons are shared by two adjacent rings (the ring is a "fused ring"), and at least one ring is an aromatic hydrocarbon , Other rings can be, for example, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and/or heterocyclic group.
  • the term “aryl” refers to a phenyl group.
  • aryl has 6 to 10 carbon atoms.
  • aromatic heterocyclic ring refers to monocyclic, bicyclic and polycyclic aromatic groups having 3 to 12 total atoms in the ring structure, including one or more heteroatoms such as nitrogen, oxygen or sulfur.
  • exemplary aromatic heterocyclic groups include azaindolyl, benzo(b)thienyl, benzimidazolyl, benzofuranyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzene O-triazolyl, benzoxadiazolyl, furyl, imidazolyl, imidazopyridyl, indolyl, indolinyl, indazolyl, isoindolinyl, isoxazolyl, isothiazolyl , Isoquinolinyl, oxazolyl, purinyl, pyranyl, pyrazinyl, pyrazolyl, pyridinyl, pyrimidinyl,
  • aromatic heterocyclic ring also includes a polycyclic ring system having two or more rings, in which two or more carbons are shared by two adjacent rings (the ring is a "fused ring"), in which at least one ring It is an aromatic group having one or more heteroatoms in the ring structure, for example, other rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and/or heterocyclic group.
  • aromatic heterobicyclic ring refers to a bicyclic aromatic group having 3 to 12 total atoms in the ring structure, including one or more heteroatoms such as nitrogen, oxygen, or sulfur.
  • exemplary heterobicyclic rings include azaindolyl, benzo(b)thienyl, benzimidazolyl, benzofuranyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzo Triazolyl, benzoxadiazolyl, imidazopyridyl, indolyl, indolinyl, indazolyl, isoindolinyl, isoxazolyl, isothiazolyl, isoquinolinyl, pyrrole And [2,3-d]pyrimidinyl, pyrrolo[2,3-b]pyridine, pyrazolo[3,4-d]pyrimidinyl, quinolinyl, quinazoline, etc.
  • aromatic heterocyclic ring also includes a polycyclic ring system having two or more rings, in which two or more carbons are shared by two adjacent rings (the ring is a "fused ring"), in which at least one ring It is an aromatic group having one or more heteroatoms in the ring structure, for example, other rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and/or heterocyclic group.
  • the term "monocyclic or fused polycyclic non-aromatic ring system” refers to excluding all ring systems other than aromatic or heteroaromatic, including saturated or unsaturated monocyclic or fused polycyclic (including bicyclic ), which may optionally contain 0-3 heteroatoms independently selected from N, O, and S.
  • the heteroatom-containing group does not have aromaticity and has 5-10 total atoms in the ring structure.
  • the "monocyclic or fused polycyclic non-aromatic ring system” may include but is not limited to: 5-membered monocyclic ring, such as tetrahydrofuryl, tetrahydrothienyl, dioxolyl, pyrrolidinyl, imidazolidine Group, pyrazolidinyl, oxazolidinyl, pyrrolinyl; or 6-membered monocyclic ring, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithiaalkyl, thiomorpholinyl, piperazine Group or trithiaalkyl; or 7-membered but ring, such as diazeppanyl.
  • 5-membered monocyclic ring such as tetrahydrofuryl, tetrahydrothienyl, dioxolyl, pyrrolidinyl, imidazolidine Group, pyrazolidinyl, ox
  • the "monocyclic or fused polycyclic non-aromatic ring system” may be benzo-fused, such as but not limited to dihydroisoquinolinyl.
  • the "monocyclic or fused polycyclic non-aromatic ring system” may be bicyclic, such as but not limited to azabicyclo[3.1.0]hexane, hexahydrocyclopenta[c]pyrrole-2(1H) -Yl ring, hexahydropyrrolo[1,2-a]pyrazine-2(1H)-yl ring.
  • substituted means that any one or more hydrogen atoms on a specific atom is replaced by a substituent, and may include deuterium and hydrogen variants, as long as the valence of the specific atom is normal and after substitution The compound is stable.
  • it means that two hydrogen atoms are replaced.
  • Carbonyl substitution does not occur on aromatic groups.
  • optionally substituted by the following substituents means that it may be substituted or unsubstituted. Unless otherwise specified, the type and number of the substituents may be arbitrary on the basis that they can be chemically realized.
  • the wavy line that intersects the bond in the chemical structure indicates the attachment point of the bond, and its wavy bond in the chemical structure intersects the rest of the molecule.
  • salts include salts derived from inorganic or organic acids including but not limited to hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, phosphoric acid , Formic acid, acetic acid, lactic acid, maleic acid, fumaric acid, succinic acid, tartaric acid, glycolic acid, salicylic acid, citric acid, methanesulfonic acid, benzenesulfonic acid, benzoic acid, malonic acid, trifluoroacetic acid, three Chloroacetic acid, naphthalene-2-sulfonic acid and other acids.
  • the pharmaceutically acceptable salt includes the case where the molar ratio of the compound base to the salt is not 1:1.
  • the salt may contain two molecules of organic acid or inorganic acid per base molecule.
  • the salt may contain 1/2 molecule of organic acid or inorganic acid per base molecule.
  • solvate refers to the physical association of a compound of the present application with one or more solvent molecules; this physical association involves various degrees of ionic and covalent bonds, including hydrogen bonds; in some cases below, for example, when one or more solvent molecules are introduced into the crystal lattice of a crystalline solid, the solvate will be able to be separated; “solvate” encompasses both solution phase and isolatable solvate; non-limiting examples of suitable solvents include But not limited to isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, ethanolamine, etc.; "hydrate” is a solvate in which the solvent molecule is H 2 O.
  • the structure described herein is also meant to include all isomeric (such as enantiomers, diastereomers, and geometric (or conformational)) forms of the structure; for example, each asymmetry
  • all tautomeric forms of the compounds of the present invention belong to the scope of the present invention.
  • the present invention includes all pharmaceutically acceptable isotopically-labeled compounds, that is, one or more atoms in the compound of formula (I) are replaced by atoms having the same atomic number but usually different from the naturally-occurring atomic mass or mass number.
  • isotopes in the compounds of the present invention include, but are not limited to: isotopes of hydrogen, such as 2 H and 3 H; isotopes of carbon, such as 11 C, 13 C, and 14 C; isotopes of chlorine, such as 36 Cl; isotopes of fluorine, For example 18 F; isotopes of iodine, such as 123 I and 125 I; isotopes of nitrogen, such as 13 N and 15 N; isotopes of oxygen, such as 15 O, 17 O, and 18 O; isotopes of phosphorus, such as 32 P; and sulfur Isotope of, for example 35 S.
  • isotopes of hydrogen such as 2 H and 3 H
  • isotopes of carbon such as 11 C, 13 C, and 14 C
  • isotopes of chlorine such as 36 Cl
  • isotopes of fluorine For example 18 F
  • isotopes of iodine such as 123 I and
  • prodrug refers to a compound that can be converted into a compound of the invention in vivo by means of metabolism (for example, by hydrolysis, reduction, or oxidation).
  • pharmaceutically acceptable carrier refers to any preparation or carrier medium that can deliver an effective amount of the active compound of the present invention, does not interfere with the biological activity of the active compound, and has no toxic or side effects to the host or patient, including any preparation or carrier medium known to those of ordinary skill in the art Solvents, dispersion media, coating materials, surfactants, antioxidants, preservatives (such as antibacterial agents, antifungal agents), isotonic agents, absorption delaying agents, salts, preservatives, drugs, drug stabilizers, viscosity Mixtures, excipients, disintegrants, lubricants, sweeteners, correctives, dyes, etc. and combinations thereof
  • excipient is used herein to describe any ingredient other than the compound of the present invention, which can impart functionality (such as drug release rate control) and/or non-functionality (such as processing aids or Thinner).
  • functionality such as drug release rate control
  • non-functionality such as processing aids or Thinner.
  • the choice of excipients largely depends on factors such as the specific mode of administration, the effect of the excipients on solubility and stability, and the nature of the dosage form.
  • Exemplary excipients may include, but are not limited to, buffers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacity agents, glidants, processing aids, One or more of colorants, sweeteners, fragrances, flavors, diluents and other known additives.
  • the compounds of the present invention can be administered in any convenient use form, such as tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches and the like.
  • Such compositions may contain conventional components in pharmaceutical preparations, such as diluents, carriers, pH adjusters, sweeteners, fillers, and additional active agents.
  • the active compounds of the present invention can also be formulated into sustained-release dosage forms.
  • the compounds of the present invention can be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal and dural Externally and intranasally, and, if necessary, for topical treatment, intralesional administration.
  • Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal, intracerebral, intraocular, intralesional, or subcutaneous administration.
  • the compound of the present invention may be in a form suitable for injection into the intraocular region of a patient, especially, in a form suitable for intravitreal injection Apply.
  • therapeutically effective amount of the compound of the present invention refers to the amount of the compound of the present invention that causes a patient's biological or medical response, such as reducing or inhibiting enzyme or protein activity or improving symptoms, alleviating symptoms, slowing down or delaying disease progression Or prevent disease, etc.
  • therapeutically effective amount refers to the amount of the compound of the present invention that is capable of at least partially reducing or inhibiting plasma kallikrein when administered to cells or tissues or non-cellular biological substances or mediators. Enzyme activity; or at least partially reduce or inhibit plasma kallikrein expression.
  • the therapeutically effective dose of the compound, pharmaceutical composition or combination depends on the patient's species, weight, age and individual condition, disease or condition, or the severity of the need for treatment.
  • the total daily dosage of the compounds and compositions of the present invention will be determined by the attending physician within the scope of reasonable medical judgment.
  • the compounds are named manually or by Chemdraw software, and the commercially available compounds use the supplier catalog name.
  • a series of tricyclic plasma kallikrein inhibitor compounds with novel structure, good activity and high selectivity are provided, which can be widely used in the prevention or treatment of diseases related to plasma kallikrein activity.
  • Figure 1 shows the fluorescein leakage scores of compounds 22, 6, 71 and 73 in a CA-1 stimulated vascular permeability model in rats;
  • Figure 2 shows the changes in retinal thickness of compounds 1, 6, 22 and 73 in a CA-1 stimulated vascular permeability model in rats;
  • Figure 3 shows the toxicity of some compounds to ARPE19 of retinal pigment epithelial cells.
  • Methanesulfonyl chloride (2.85mL, 36.794mmol) was added dropwise to (4-((3-bromo-5-methyl-1H-pyrazol-1-yl)methyl)-2-nitrobenzyl alcohol (7.5g , 22.996mmol) and triethylamine (6.4mL, 45.992mmol) in dichloromethane (70mL) solution at 35°C for 4h.
  • reaction solution was diluted with ice water (200mL) and dichloromethane (150mL ⁇ 3) Extract, combine the organic layers, wash with saturated sodium chloride aqueous solution (100mL ⁇ 3), dry with anhydrous sodium sulfate, filter, and evaporate the filtrate under reduced pressure to obtain 4-((3-bromo-5-methyl-1H -Pyrazol-1-yl)methyl)-2-nitrobenzyl alcohol methanesulfonate, used directly in the next reaction without purification.
  • reaction solution was Quench with aqueous ammonium chloride solution (10 mL) at 20°C, extract with dichloromethane (80 mL ⁇ 2), combine the organic layers, wash with saturated aqueous sodium chloride solution (50 mL ⁇ 2), dry with anhydrous sodium sulfate, filter, and reduce the filtrate. It was evaporated to dryness to obtain a white solid with a crude yield of 93.2%. It was directly used in the next reaction without purification, ESI-MS (m/z): 474.0 [M+H] + .
  • reaction solution was quenched with ammonium chloride aqueous solution (100mL) at 0°C, extracted with ethyl acetate (200mL ⁇ 2), and combined The organic layer was washed with saturated aqueous sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure.
  • Methanesulfonyl chloride (0.786mL, 10.146mmol) was added dropwise to 4-((1H-pyrazol-1-yl)methyl)-2-nitrobenzyl alcohol (1.5g, 6.431mmol) and triethylamine (1.79 mL, 12.862mmol) in dichloromethane (15mL) solution, react at 35°C for 8h.
  • the operation process is the same as in Example 2, except that the starting material 1H-pyrazole in step a is replaced by 3-azabicyclo[3.1.0]hexan-2-one, potassium carbonate is replaced by NaH, and acetonitrile is replaced by tetrahydrofuran.
  • Methyl 4-bromo-2-nitrobenzoate (5.0g, 19.228mmol), aniline (1.97g, 21.151mmol), cesium carbonate (9.40g, 28.842mmol), dicyclohexyl[3,6-dimethyl Oxy-2',4',6'-triisopropyl[1,1'-biphenyl]-2-yl]phosphine (BrettPhos, 516mg, 0.961mmol) and Pd 2 (dba) 3 (880mg, 0.961 mmol) was added to the reaction flask, argon replaced the air in the reaction flask and then anhydrous tetrahydrofuran (100mL) was added.
  • the operation process is the same as in Example 2, except that the 1H-pyrazole in step a is replaced by 4-methyl-1H-pyrazole, and the 5-(aminomethyl)-4,6-dimethyl in step i is replaced by 4-methyl-1H-pyrazole.
  • the operation process is the same as in Example 2, except that the 1H-pyrazole in step a is replaced by 4-fluoro-1H-pyrazole, and the 5-(aminomethyl)-4,6-lutidine in step i is replaced by 4-fluoro-1H-pyrazole.
  • Diaza Tert-Butyl-2-carboxylate (350mg, 0.890mmol) and 5N hydrochloric acid (0.71mL) were added to the reaction flask, heated to 45°C and reacted for 6h. After the reaction, the solvent was evaporated under reduced pressure, and the crude product obtained was directly used in the next reaction , The yield is 86.6%; ESI-MS (m/z): 338.1 [M+H] + .
  • step b in implementation 22 Take 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4] Diaza -2-carboxylic acid and (1H-pyrrole[2,3-b]pyridin-5-yl)methylamine as raw materials, the operation process is the same as step b in implementation 22 to obtain N-((1H-pyrrole[2,3 -b]pyridin-5-yl)methyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole [1,2-a][1,4] Diaza -2-carboxamide; ESI-MS (m/z): 453.3 [M+H] + .
  • Methanesulfonyl chloride (6.21mL, 54.209mmol) was added dropwise to (4-((3-bromo-5-methyl-1H-pyrazol-1-yl)methyl)-2-nitrobenzyl alcohol (8.0g , 24.529mmol) and triethylamine (10.2mL, 73.587mmol) in dichloromethane (100mL) solution at 25°C for 12h.
  • reaction solution is diluted with ice water (100mL) and dichloromethane (100mL) ⁇ 3) Extract, combine the organic layers, wash with saturated aqueous sodium chloride solution (80mL ⁇ 3), dry with anhydrous sodium sulfate, filter, and evaporate the filtrate under reduced pressure to obtain 3-bromo-1-(4-(chloromethyl) -3-nitrobenzyl)-5-methyl-1H-pyrazole, used directly in the next reaction without purification; ESI-MS (m/z): 344.0 [M+H] + .
  • reaction solution is heated with ice Dilute with water (100mL), extract with ethyl acetate (100mL ⁇ 2), combine the organic layers, wash with saturated sodium chloride aqueous solution (80mL ⁇ 3), dry with anhydrous sodium sulfate, filter, and evaporate the filtrate under reduced pressure to obtain 1- (4-((3-Bromo-5-methyl-1H-pyrazol-1-yl)methyl)-2-nitrobenzyl)-1H-pyrrole-2,4-dicarboxylate, without purification It was directly used in the next reaction; ESI-MS (m/z): 519.1 [M+H] + .
  • Step ef in 2 gives 8-((1H-pyrazol-1-yl)methyl)-11-oxo-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a ][1,4]diazepine Ethyl-2-carboxylate; ESI-MS (m/z): 351.2 [M+H] + .
  • the reaction mixture Add dichloromethane (100mL) to dilute, wash with 1N hydrochloric acid aqueous solution (100mL), saturated sodium chloride aqueous solution (100mL) successively, dry with anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure.
  • the ester mixed solvent (9:1, v/v, 50 mL) was slurried, filtered, and dried under reduced pressure to obtain an off-white solid with a yield of 90.5%; ESI-MS (m/z): 709.4 [M+H] + .
  • the method reported by HCHemker et al. (Handbook of Synthetic Substrates) was used with some improvements.
  • the method reported by HCHemker et al. (Handbook_of_Synthetic_Subst) was used with some improvements.
  • Add 10 ⁇ L of 15nM human coagulation factor XIa (commercially available from Enzyme Research Laboratories) and 5 ⁇ L of the test compound solution to the 384 microwell plate. After adding and mixing, incubate at 37°C for 15min, and add 5 ⁇ L of 400 ⁇ M reaction substrate N to each well.
  • 3nM human coagulation factor Xa commercially available from Enzyme Research Laboratories
  • 5 ⁇ L of the test compound solution was added to the 384 microwell plate. After adding and mixing, incubate at 37°C for 15min, and add 5 ⁇ L of 400 ⁇ M reaction substrate N to each well.
  • Human and animal microsomes were purchased from Corning or Xenotech and stored in a refrigerator at -80°C.
  • NADPH reduced nicotinamide adenine dinucleotide phosphate
  • T60 incubation plate and NCF60 incubation plate prepare two 96-well incubation plates, named T60 incubation plate and NCF60 incubation plate, respectively.
  • test substance or control compound solution After the pre-incubation, add 5 ⁇ L of the test substance or control compound solution to the T60 incubation plate and the NCF60 incubation plate, and mix well. Add 50 ⁇ L of potassium phosphate buffer to each well of the NCF60 incubation plate to start the reaction; add 180 ⁇ L of stop solution (containing 200ng/mL tolbutamide and 200ng/mL labetalol in acetonitrile solution) and 6uL of NADPH regeneration system working solution to the T0 stop plate , Take 54 ⁇ L sample from T60 incubation plate to T0 stop plate (T0 sample generation). Add 44 ⁇ L of NADPH regeneration system working solution to each well of the T60 incubation plate to start the reaction.
  • stop solution containing 200ng/mL tolbutamide and 200ng/mL labetalol in acetonitrile solution
  • NADPH regeneration system working solution Take 54 ⁇ L sample from T60 incubation plate to T0 stop plate (T0 sample generation).
  • the final concentration of the compound, testosterone, diclofenac and propafenone in the reaction is 1 ⁇ M
  • the concentration of liver microsomes is 0.5mg/mL
  • the final concentration of DMSO and acetonitrile in the reaction system are 0.01% (v/v) and 0.99% (v/v), respectively.
  • stop solution acetonitrile solution containing 200ng/mL tolbutamide and 200ng/mL labetalol
  • All sample plates were shaken well and centrifuged at 3220 ⁇ g for 20 minutes, and then 80 ⁇ L of supernatant was diluted into 240 ⁇ L of pure water from each well for liquid chromatography tandem mass spectrometry analysis.
  • Liquid chromatography tandem mass spectrometry analyzes all samples, using in vitro metabolic half-life (T 1/2 , min) and intrinsic clearance (CL int ) as indicators to investigate the metabolic stability of the compounds. The results are shown in Table 5. :
  • Mass spectrometer Sciex API4000TM Ultra high performance liquid chromatography LC30-AD, Shimadzu; autosampler SIL30-ACMP, Shimadzu
  • test compound is dissolved in DMSO and prepared into a 10mM DMSO stock solution;
  • Working concentration preparation Dissolve an appropriate amount of K 2 HPO 4 ⁇ 3H 2 O (analytical purity) and KH 2 PO 4 (analytical purity) in an appropriate amount of ultrapure water, and adjust the pH to 7.40 ⁇ with H 3 PO 4 or KOH 0.10, formulated into a buffer with a concentration of 100 mM, to obtain potassium phosphate buffer (PB).
  • K 2 HPO 4 ⁇ 3H 2 O analytical purity
  • KH 2 PO 4 analytical purity
  • Weight at the time of purchase 250 ⁇ 20g
  • Group design blank control group (normal saline + normal saline Sham group), model control group (normal saline + CA-I group), test product group (compounds 22, 6, 71, 73, a total of 6 groups); , CA-I is rat carbonic anhydrase 1 (purchased from Sigma-Aldrich).
  • Compound tropikamide eye drops were used to dilate pupils, and 1% pentobarbital sodium was used for intraperitoneal injection of general anesthesia.
  • the rats were fixed on the operating table under a microscope, and tobramycin and dexamethasone eye drops were used to flush the conjunctival sac.
  • the needle was inserted vertically at 1mm of the corneoscleral limbus, and 2uL normal saline or test compound (compounds 22, 6, 71, 73) was injected into the vitreous body. 30 minutes later, 2uL normal saline or CA-I (60ng/eye) was injected into the vitreous for the second time. ) Modeling (monocular).
  • SD rats purchased from Zhejiang Weitong Lihua Laboratory Animal Technology Co., Ltd.
  • Group design blank control group (normal saline + normal saline Sham group), model control group (normal saline + CA-I group), test product group (compounds 1, 6, 22, 73, a total of 6 groups); , CA-I is rat carbonic anhydrase 1 (purchased from Sigma-Aldrich).
  • the animals were anesthetized by intramuscular injection of Zoletil and Xylazine at doses of 5-15 mg/kg and 2-12 mg/kg, and Oxybuprocaine eye drops were instilled into the conjunctival sac. Table anesthesia. Place the animal on the operating table under a stereo microscope, and inject 5uL/eye normal saline or test compound (compounds 1, 6, 22, 73) into the vitreous body. After 30 minutes, inject the second time 5uL/eye normal saline or CA-I (100ng/eye) model (monocular). Intramuscular injection of Idzoxan Hydrochloride injection made the animals wake up normally. They were then returned to their cages and antibiotic eye drops were given 3 times a day.
  • OCT optical coherence tomography
  • OCT Inspection method Perform an OCT (Blue Spectrails OCT, Heidelberg) inspection on the eyes of all animals before injection, and then perform an OCT inspection about 48 hours after modeling, and record the retinal thickness ( ⁇ M) of the two OCT inspections.
  • OCT Blue Spectrails OCT, Heidelberg

Abstract

Provided is a tricyclic plasma kallikrein inhibitor compound having a novel structure, good activity and high selectivity, which can be widely used in the prevention or treatment of diseases related to a plasma kallikrein activity.

Description

作为血浆激肽释放酶抑制剂的三环类化合物及其用途Tricyclic compounds as plasma kallikrein inhibitors and their uses 技术领域Technical field
本发明涉及对血浆激肽释放酶具有选择性抑制活性的三环类化合物及其用途。The present invention relates to tricyclic compounds with selective inhibitory activity on plasma kallikrein and uses thereof.
背景技术Background technique
血浆激肽释放酶(plasma kallikrein,PK)属丝氨酸蛋白酶家族的一员，最早是在哺乳动物的血浆中被发现。它由位于4q35染色体上单个基因(KLKB1)编码，主要在肝脏中合成并以血浆激肽释放酶原(PPK)形式大量存在于血液循环中，PPK进一步通过凝血因子XIIa剪切其内在的Arg-IIe键激活转化成PK(Yousef,G.M.et al.An overview of the kallikrein gene families in humans and other species:Emerging candidate tumour markers.Clin.Biochem.2003,36,443-452)。PK是激肽释放酶-激肽系统(KKS)的一个关键酶，可作用于高分子量激肽原(KH)，从而使其活化释放小分子缓激肽(BK)，进而通过作用于缓激肽受体参与凝血、纤维蛋白溶解、补体激活以及炎症发生等生物学过程。Plasma kallikrein (PK) is a member of the serine protease family and was first discovered in the plasma of mammals. It is encoded by a single gene (KLKB1) located on the 4q35 chromosome. It is mainly synthesized in the liver and is abundantly present in the blood circulation in the form of plasma kallikrein (PPK). PPK further cleaves its intrinsic Arg- by coagulation factor XIIa. IIe key activation is transformed into PK (Yousef, GMet al. An overview of the kallikrein gene families in humans and other species: Emerging candidates tumor markers. Clin. Biochem. 2003, 36, 443-452). PK is a key enzyme of the kallikrein-kinin system (KKS), which can act on high molecular weight kininogen (KH) to activate the release of small-molecule bradykinin (BK), and then act on bradykinin Peptide receptors are involved in biological processes such as blood coagulation, fibrinolysis, complement activation and inflammation.
近年来，随着对血浆激肽释放酶的基因学、分子学和药理学的研究更加深入，人们对其生理和病理角色有了深入的认识。研究表明，血浆激肽释放酶与炎症性疾病、肿瘤、心血管疾病、肾病、中枢神经系统疾病、视网膜病以及糖尿病视网膜疾病等多种疾病密切相关(Costa-Neto,C.M.et al.Participation of kallikrein-kinin system in different pathologies.Int.Immunopharmacol.2008,8,135-142)。例如遗传性血管水肿(HAE)，它是一种常染色体显性遗传性，主要是由于患者体内C1-INH缺乏，导致其对血浆激肽释放酶的抑制作用减弱，不受控制的激活KKS系统，释放血管活性物质，使血管通透性增加引起典型的肿胀(Farkas,H.Orphan drugs for the treatment of hereditary angioedema.Expert Opinion on Orphan Drugs,2015,1,141-156)。目前已有艾卡拉肽和Lanadelumab等大分子血浆激肽释放酶抑制剂上市在临床上用于治疗遗传性血管水肿，疗效显著。又例如，在糖尿病黄斑水肿患者眼部玻璃体中，发现其KKS系统过度激活，导致视网膜血管通透性增加和视网膜增厚。近年来已公开多项资料表明血浆激肽释放酶抑制剂可以降低视网膜血管通透性，用于治疗糖尿病视网膜疾病和糖尿病黄斑水肿(Feener,E.P.Plasma kallikrein and diabetic macular edema.Curr.Diab.Rep.2010,10,270-275；Liu J.et al.Plasma kallikrein-kinin system and diabetic retinopathy.Biol.Chem.2013,394,319-328)。目前已有小分子(NCT03466099)和多肽类(NCT03511898)血浆激肽释放酶抑制剂用于治疗糖尿病黄斑水肿进入临床研究。In recent years, as the genetics, molecular and pharmacology of plasma kallikrein have been studied more deeply, people have gained in-depth understanding of its physiological and pathological roles. Studies have shown that plasma kallikrein is closely related to inflammatory diseases, tumors, cardiovascular diseases, nephropathy, central nervous system diseases, retinopathy, and diabetic retinopathy. Participation of kallikrein -kinin system in different pathologies.Int.Immunopharmacol.2008,8,135-142). For example, hereditary angioedema (HAE), which is an autosomal dominant inheritance, is mainly due to the lack of C1-INH in the patient's body, which leads to a weakened inhibitory effect on plasma kallikrein and uncontrolled activation of the KKS system , Release vasoactive substances, increase vascular permeability and cause typical swelling (Farkas, H. Orphan drugs for the treatment of hereditary angioedema. Expert Opinion on Orphan Drugs, 2015, 1, 141-156). At present, large-molecule plasma kallikrein inhibitors such as Icaratide and Lanadelumab have been marketed clinically for the treatment of hereditary angioedema, with significant curative effects. For another example, in the ocular vitreous of patients with diabetic macular edema, it is found that the KKS system is over-activated, which leads to increased retinal blood vessel permeability and thickening of the retina. In recent years, a number of data have been published showing that plasma kallikrein inhibitors can reduce retinal vascular permeability, and are used to treat diabetic retinopathy and diabetic macular edema (Feener, EPPlasma kallikrein and diabetic macular edema.Curr.Diab.Rep. 2010, October, 270-275; Liu J.et al. Plasma Kallikrein-kinin system and diabetic retinopathy. Biol. Chem. 2013, 394, 319-328). At present, small molecules (NCT03466099) and peptides (NCT03511898) plasma kallikrein inhibitors have been used in the treatment of diabetic macular edema into clinical research.
然而，现有报道的多肽和小分子类血浆激肽释放酶抑制剂又存在诸多限制。如艾卡拉肽被报道存在引起过敏性反应的风险(Banta,E.et al.Overview of ecallantide in the treatment of hereditary angioedema types I and II.Therapy.2010,7,565-571)。而现有报道的绝大部分小分子血浆激肽释放酶抑制剂具有高度极性和可离子化的胍基或脒基官能团，这被认为难以透过生物膜导致口服生物利用度非常差(Tamie,J.et al.ASP-634:an oral drug candidate for diabetic macular edema.ARVO Annual Meeting Abstract,2012,53,2240)。Biocryst开发了口服血浆激肽酶抑制剂BCX4161(Collis,P.et al.,BCX4161,an oral kallikrein inhibitor:safety and pharmacokinetic results of a Phase 1study in healthy volunteers.J.Allergy Clin.Immunol 2014,133,Volume 133,Issue 2,Supplement,2014,AB39)，但是其临床给药剂量为400mg每次，每日三次给药。结构中苯甲脒或胍基虽构成BCX4161的药效基团，但其较差的理化性质可能影响了药物的生物利用度，因此需以较大剂量给药。此外现有的血浆激肽释放酶抑制剂还存在 对相关酶如KLK1、凝血酶和其他丝氨酸蛋白酶选择性较差的问题。至今还没有小分子的血浆激肽释放酶抑制剂被批准上市。因此，研究新型、作用力更强、副作用更小的选择性血浆激肽释放酶抑制剂仍有需求。However, the peptides and small molecule plasma kallikrein inhibitors reported currently have many limitations. For example, Ikaratide is reported to have the risk of causing allergic reactions (Banta, E. et al. Overview of ecallantide in the treatment of hereditary angioedema types I and II. Therapy. 2010, 7,565-571). However, most of the small-molecule plasma kallikrein inhibitors reported currently have highly polar and ionizable guanidino or amidino functional groups, which are considered to be difficult to penetrate biofilms and lead to poor oral bioavailability (Tamie ,J.et al.ASP-634:an oral drug candidate for diabetes macular edema.ARVO Annual Meeting Abstract,2012,53,2240). Biocryst developed the oral plasma kallikrein inhibitor BCX4161 (Collis, P.et al., BCX4161, an oral kallikrein inhibitor: safety and pharmacokinetic results of a Phase 1 study in healthy volunteers.J.Allergy Clin.Immunol 2014, 133, 133, Issue 2, Supplement, 2014, AB39), but its clinical administration dose is 400 mg each time, three times a day. Although the benzamidine or guanidine group constitutes the pharmacodynamic group of BCX4161 in the structure, its poor physical and chemical properties may affect the bioavailability of the drug, so it needs to be administered in a larger dose. In addition, the existing plasma kallikrein inhibitors also have the problem of poor selectivity to related enzymes such as KLK1, thrombin and other serine proteases. So far, no small molecule plasma kallikrein inhibitor has been approved for marketing. Therefore, there is still a need to study new types of selective plasma kallikrein inhibitors with stronger effects and fewer side effects.
发明内容Summary of the invention
本发明涉及血浆激肽释放酶抑制剂化合物、所述化合物的药物组合物、所述化合物的制药用途以及化合物的治疗方法。The present invention relates to a plasma kallikrein inhibitor compound, a pharmaceutical composition of the compound, the pharmaceutical use of the compound, and a treatment method of the compound.
在某一方面，本发明提供式(I)的化合物，其药学上可接受的盐、溶剂化物、立体异构体或前药：In a certain aspect, the present invention provides a compound of formula (I), a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof:
Figure PCTCN2020137640-appb-000001
Figure PCTCN2020137640-appb-000001
其中，among them,
A选自5元或6元芳环或芳杂环，所述芳杂环含有1-3个选自N、O和S的杂原子，所述芳环或芳杂环任选地被下述取代基取代：卤素、烷基、烷氧基、卤代烷基、OH、CN、COOR 1、CONR 1R 2、NR 1R 2、NR 1COR 2、(CH 2) 1-3NR 1R 2、(CH 2) 1-3OR 1和C(R 1)(R 2)NH 2； A is selected from a 5-membered or 6-membered aromatic ring or an aromatic heterocyclic ring, the aromatic heterocyclic ring containing 1-3 heteroatoms selected from N, O and S, and the aromatic ring or aromatic heterocyclic ring is optionally Substituent substitution: halogen, alkyl, alkoxy, haloalkyl, OH, CN, COOR 1 , CONR 1 R 2 , NR 1 R 2 , NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1-3 OR 1 and C(R 1 )(R 2 )NH 2 ;
或者A选自稠合的6，5-或6，6-芳杂双环，所述芳杂双环含有N和任选的另外独立地1-2个选自N、O和S的杂原子，所述芳杂双环任选地被下述取代基取代：卤素、烷基、烷氧基、卤代烷基、OH、CN、COOR 1、CONR 1R 2、NR 1R 2、NR 1COR 2、(CH 2) 1-3NR 1R 2、(CH 2) 1-3OR 1和-C(R 1)(R 2)NH 2； Or A is selected from a fused 6,5- or 6,6-aromatic heterobicyclic ring containing N and optionally additionally independently 1-2 heteroatoms selected from N, O and S, so The heterobicyclic ring is optionally substituted by the following substituents: halogen, alkyl, alkoxy, haloalkyl, OH, CN, COOR 1 , CONR 1 R 2 , NR 1 R 2 , NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1-3 OR 1 and -C(R 1 )(R 2 )NH 2 ;
L 1选自(CR 3R 4) m或(CR 3R 4) mO，其中m为0、1、2； L 1 is selected from (CR 3 R 4 ) m or (CR 3 R 4 ) m O, where m is 0, 1, 2;
X 1和X 2为CR 5或N，且不同时为N；其中R 5选自H、OH、卤素、烷基、烷氧基、卤代烷基、环烷基、CN、COOR 1、CONR 1R 2、NR 1R 2、NR 1COR 2、(CH 2) 1-3NR 1R 2、(CH 2) 1-3OR 1和-C(R 1)(R 2)NH 2； X 1 and X 2 are CR 5 or N, and are not N at the same time; wherein R 5 is selected from H, OH, halogen, alkyl, alkoxy, haloalkyl, cycloalkyl, CN, COOR 1 , CONR 1 R 2. NR 1 R 2 , NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1-3 OR 1 and -C(R 1 )(R 2 )NH 2 ;
-C-D-选自-NH-CH 2-、-N＝CH-、-NCH 3-CH 2-、-NHCO-、-CH＝CH-或-CH 2-CH 2-； -CD- is selected from -NH-CH 2 -, -N=CH-, -NCH 3 -CH 2 -, -NHCO-, -CH=CH- or -CH 2 -CH 2 -;
L 2选自键、(CH 2) n、(CH 2) nNH、(CH 2) nO，其中n为1或2； L 2 is selected from bond, (CH 2 ) n , (CH 2 ) n NH, (CH 2 ) n O, where n is 1 or 2;
B选自5-6元芳环或者芳杂环，其中所述芳杂环含有1-3选自N、O或S的杂原子，任选地，所述芳环或者芳杂环被烷基、烷氧基、OH、卤素、CN、酮基、COOR 1、CONR 1R 2、NR 1R 2、NR 1COR 2、(CH 2) 1-3NR 1R 2、(CH 2) 1-3OR 1和-C(R 1)(R 2)NH 2取代； B is selected from a 5-6 membered aromatic ring or an aromatic heterocyclic ring, wherein the aromatic heterocyclic ring contains 1-3 heteroatoms selected from N, O or S. Optionally, the aromatic ring or aromatic heterocyclic ring is alkylated by an alkyl group. , Alkoxy, OH, halogen, CN, ketone, COOR 1 , CONR 1 R 2 , NR 1 R 2 , NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1- 3 OR 1 and -C(R 1 )(R 2 )NH 2 are substituted;
或者B选自饱和的或不饱和的5-10元单环或稠合多环非芳族环系，所述非芳族环系任选地含有0-3个选自N、O和S的杂原子，所述非芳族环系任选地被下述取代基取代：卤素、烷基、烷氧基、卤代烷基、酮基、OH、CN、COOR 1、CONR 1R 2、NR 1R 2、NR 1COR 2、(CH 2) 1-3NR 1R 2、(CH 2) 1-3OR 1和-C(R 1)(R 2)NH 2； Or B is selected from a saturated or unsaturated 5-10 membered monocyclic or fused polycyclic non-aromatic ring system, the non-aromatic ring system optionally contains 0-3 selected from N, O and S Heteroatoms, the non-aromatic ring system is optionally substituted by the following substituents: halogen, alkyl, alkoxy, haloalkyl, keto, OH, CN, COOR 1 , CONR 1 R 2 , NR 1 R 2. NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1-3 OR 1 and -C(R 1 )(R 2 )NH 2 ;
R 1和R 2独立地选自H和烷基，或者R 1和R 2与它们所连接的碳一起形成环烷基； R 1 and R 2 are independently selected from H and alkyl, or R 1 and R 2 together with the carbon to which they are attached form a cycloalkyl;
R 3和R 4独立地选自H和烷基，或者R 3和R 4与它们所连接的碳一起形成环烷基。 R 3 and R 4 are independently selected from H and alkyl, or R 3 and R 4 together with the carbon to which they are attached form a cycloalkyl.
在另一实施方案中，在式(I)的化合物中，A选自
Figure PCTCN2020137640-appb-000002
其中，X 3为CR 10或N，R 6、R 7、R 8、R 9、R 10独立地选自氢、卤素、烷基、烷氧基、卤代烷基、OH、CN、COOR 1、CONR 1R 2、NR 1R 2、NR 1COR 2、(CH 2) 1-3NR 1R 2、(CH 2) 1-3OR 1和C(R 1)(R 2)NH 2；R 1和R 2独立地选自H和烷基，或者R 1和R 2与它们所连接的碳一起形成环烷基。 In another embodiment, in the compound of formula (I), A is selected from
Figure PCTCN2020137640-appb-000002
Wherein, X 3 is CR 10 or N, R 6 , R 7 , R 8 , R 9 , R 10 are independently selected from hydrogen, halogen, alkyl, alkoxy, haloalkyl, OH, CN, COOR 1 , CONR 1 R 2 , NR 1 R 2 , NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1-3 OR 1 and C(R 1 )(R 2 )NH 2 ; R 1 And R 2 are independently selected from H and an alkyl group, or R 1 and R 2 together with the carbon to which they are attached form a cycloalkyl group.
在另一实施方案中，在式(I)的化合物中，R 8为NH 2或者C(R 1)(R 2)NH 2，其中R 1和R 2独立地选自H和C 1-3的烷基，或者R 1和R 2与它们所连接的碳一起形成3元或4元环烷基。 In another embodiment, in the compound of formula (I), R 8 is NH 2 or C(R 1 )(R 2 )NH 2 , wherein R 1 and R 2 are independently selected from H and C 1-3 The alkyl group, or R 1 and R 2 together with the carbon to which they are attached form a 3-membered or 4-membered cycloalkyl group.
在另一实施方案中，在式(I)的化合物中，R 8为NH 2。 In another embodiment, in the compound of formula (I), R 8 is NH 2 .
在另一实施方案中，在式(I)的化合物中，R 8为CH 2NH 2。 In another embodiment, in the compound of formula (I), R 8 is CH 2 NH 2 .
在另一实施方案中，在式(I)的化合物中，R 8为C(R 1)(R 2)NH 2，其中R 1和R 2与它们所连接的碳一起形成环丙基。 In another embodiment, in the compound of formula (I), R 8 is C(R 1 )(R 2 )NH 2 , wherein R 1 and R 2 together with the carbon to which they are attached form a cyclopropyl group.
在另一实施方案中，在式(I)的化合物中，R 6、R 7、R 9、R 10独立地选自H、卤素、烷基、烷氧基、卤代烷基。 In another embodiment, in the compound of formula (I), R 6 , R 7 , R 9 , R 10 are independently selected from H, halogen, alkyl, alkoxy, haloalkyl.
在另一实施方案中，在式(I)的化合物中，R 6、R 7、R 9、R 10立地选自H、卤素或者CH 3。 In another embodiment, in the compound of formula (I), R 6 , R 7 , R 9 , R 10 are independently selected from H, halogen, or CH 3 .
在另一实施方案中，在式(I)的化合物中，A选自：In another embodiment, in the compound of formula (I), A is selected from:
Figure PCTCN2020137640-appb-000003
Figure PCTCN2020137640-appb-000003
在另一实施方案中，在式(I)的化合物中，A选自异喹啉和1H-吡咯[2,3-b]吡啶，其中异喹啉和1H-吡咯[2,3-b]吡啶任选地被卤素、烷基、烷氧基、卤代烷基、OH、CN、COOR 1、CONR 1R 2、NR 1R 2、NR 1COR 2、(CH 2) 1-3NR 1R 2、(CH 2) 1-3OR 1和C(R 1)(R 2)NH 2取代；R 1和R 2独立地选自H和烷基，或者R 1和R 2与它们所连接的碳一起形成环烷基。 In another embodiment, in the compound of formula (I), A is selected from isoquinoline and 1H-pyrrole [2,3-b]pyridine, wherein isoquinoline and 1H-pyrrole [2,3-b] Pyridine is optionally substituted by halogen, alkyl, alkoxy, haloalkyl, OH, CN, COOR 1 , CONR 1 R 2 , NR 1 R 2 , NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1-3 OR 1 and C(R 1 )(R 2 )NH 2 substitution; R 1 and R 2 are independently selected from H and alkyl, or R 1 and R 2 and the carbon to which they are attached Together to form a cycloalkyl group.
在另一实施方案中，在式(I)的化合物中，A选自异喹啉和1H-吡咯[2,3-b]吡啶，其中异喹啉和1H-吡咯[2,3-b]吡啶任选地被卤素、烷基、烷氧基、卤代烷基和氨基取代。In another embodiment, in the compound of formula (I), A is selected from isoquinoline and 1H-pyrrole [2,3-b]pyridine, wherein isoquinoline and 1H-pyrrole [2,3-b] Pyridine is optionally substituted with halogen, alkyl, alkoxy, haloalkyl, and amino.
在另一实施方案中，在式(I)的化合物中，A选自异喹啉和1H-吡咯[2,3-b]吡啶，其中异喹啉和1H-吡咯[2,3-b]吡啶任选地被卤素、烷基、烷氧基、卤代烷基和氨基取代。In another embodiment, in the compound of formula (I), A is selected from isoquinoline and 1H-pyrrole [2,3-b]pyridine, wherein isoquinoline and 1H-pyrrole [2,3-b] Pyridine is optionally substituted with halogen, alkyl, alkoxy, haloalkyl, and amino.
在另一实施方案中，在式(I)的化合物中，A选自：In another embodiment, in the compound of formula (I), A is selected from:
Figure PCTCN2020137640-appb-000004
Figure PCTCN2020137640-appb-000004
在另一实施方案中，在式(I)的化合物中，L 1选自键、CH 2、(CH 2) 2O。 In another embodiment, in the compound of formula (I), L 1 is selected from bond, CH 2 , (CH 2 ) 2 O.
在另一实施方案中，在式(I)的化合物中，L 1选自CH 2。 In another embodiment, in the compound of formula (I), L 1 is selected from CH 2 .
在另一实施方案中，在式(I)的化合物中，X 2为CR 5，其中R 5选自H、烷基、卤代烷基和烷氧基。 In another embodiment, in the compound of formula (I), X 2 is CR 5 , wherein R 5 is selected from H, alkyl, haloalkyl, and alkoxy.
在另一实施方案中，在式(I)的化合物中，X 2为CR 5，其中R 5选自H、CH 3、CF 3、 CH 2OCH 3。 In another embodiment, in the compound of formula (I), X 2 is CR 5 , wherein R 5 is selected from H, CH 3 , CF 3 , CH 2 OCH 3 .
在另一实施方案中，在式(I)的化合物中，-C-D-选自-NH-CH 2-、-N＝CH-、-NHCO-。 In another embodiment, the compound of formula (I) in, -CD- is selected from -NH-CH 2 -, - N = CH -, - NHCO-.
在另一实施方案中，在式(I)的化合物中，L 2选自键、O、NH、CH 2、CH 2NH、CH 2O。 In another embodiment, in the compound of formula (I), L 2 is selected from bond, O, NH, CH 2 , CH 2 NH, CH 2 O.
在另一实施方案中，在式(I)的化合物中，L 2选自CH 2。 In another embodiment, in the compound of formula (I), L 2 is selected from CH 2 .
在另一实施方案中，在式(I)的化合物中，B选自：In another embodiment, in the compound of formula (I), B is selected from:
Figure PCTCN2020137640-appb-000005
Figure PCTCN2020137640-appb-000005
在另一实施方案中，式(I)的化合物选自：In another embodiment, the compound of formula (I) is selected from:
Figure PCTCN2020137640-appb-000006
Figure PCTCN2020137640-appb-000006
Figure PCTCN2020137640-appb-000007
Figure PCTCN2020137640-appb-000007
Figure PCTCN2020137640-appb-000008
Figure PCTCN2020137640-appb-000008
本发明的第二目的在于提供一种药物组合物，所述组合物包含本发明第一目的所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，和药学上可接受的赋形剂。The second object of the present invention is to provide a pharmaceutical composition comprising the compound according to the first object of the present invention, a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof, and a pharmaceutical composition. Acceptable excipients.
本发明的第三目的在于提供本发明第一目的所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，在制备用于治疗或预防涉及血浆激肽释放酶活性的疾病或病症的药物中的用途。The third object of the present invention is to provide the compound according to the first object of the present invention, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug, which is used in the treatment or prevention of plasma kallikrein. Use in medicine for active diseases or conditions.
本发明的第四目的在于提供本发明第一目的所述的化合物，其溶剂化物、互变异构体或药学上可接受的盐治疗或预防哺乳动物中涉及血浆激肽释放酶活性的疾病或病症的方法，包括向有需要的哺乳动物施用治疗有效量的本发明第一目的所述的化合物。The fourth object of the present invention is to provide the compound according to the first object of the present invention, its solvate, tautomer or pharmaceutically acceptable salt for the treatment or prevention of diseases or diseases involving plasma kallikrein activity in mammals The method of disease includes administering a therapeutically effective amount of the compound described in the first object of the present invention to a mammal in need thereof.
在另一实施方案中，涉及血浆激肽释放酶活性的疾病为炎症。In another embodiment, the disease involving plasma kallikrein activity is inflammation.
在另一实施方案中，涉及血浆激肽释放酶活性的疾病选自视力受损、糖尿病视网膜病变、糖尿病黄斑水肿、遗传性血管水肿、糖尿病、胰腺炎、脑出血、肾病、心肌病、神经病、炎性肠病、关节炎、感染性休克、低血压、癌症、成人呼吸窘迫综合征、弥散性血管内凝血、心肺旁路手术和外科手术后出血。In another embodiment, the disease involving plasma kallikrein activity is selected from the group consisting of impaired vision, diabetic retinopathy, diabetic macular edema, hereditary angioedema, diabetes, pancreatitis, cerebral hemorrhage, nephropathy, cardiomyopathy, neuropathy, Inflammatory bowel disease, arthritis, septic shock, hypotension, cancer, adult respiratory distress syndrome, diffuse intravascular coagulation, cardiopulmonary bypass surgery and bleeding after surgery.
在另一实施方案中，涉及血浆激肽释放酶活性的疾病是与糖尿病视网膜病变和糖尿病黄斑水肿相关的视网膜血管通透性疾病。In another embodiment, the disease involving plasma kallikrein activity is a retinal vascular permeability disease associated with diabetic retinopathy and diabetic macular edema.
在另一实施方案中，涉及血浆激肽释放酶活性的疾病为糖尿病黄斑水肿。In another embodiment, the disease involving plasma kallikrein activity is diabetic macular edema.
在另一实施方案中，涉及血浆激肽释放酶活性的疾病为遗传性血管水肿。In another embodiment, the disease involving plasma kallikrein activity is hereditary angioedema.
本发明的化合物可以与其他的治疗剂联合施用。联合施用疗法包括式(I)化合物和一种或多种其他的治疗剂，其他的治疗剂选自：抑制血小板衍生因子(PDGF)的治疗剂，血管内皮生长因子(VEGF)，整联蛋白α 5β 1，类固醇，抑制血浆激肽释放酶的其他治疗剂以及其他的炎症抑制剂。 The compounds of the present invention can be administered in combination with other therapeutic agents. Co-administration therapy includes a compound of formula (I) and one or more other therapeutic agents, and the other therapeutic agents are selected from: therapeutic agents that inhibit platelet-derived factor (PDGF), vascular endothelial growth factor (VEGF), integrin alpha 5 β 1 , steroids, other therapeutic agents that inhibit plasma kallikrein, and other inflammation inhibitors.
当采用联合疗法时，本发明化合物和所述联用的治疗剂可以存在于相同或不同的药物组合物中，并且可以单独地、顺序地或同时地施用。When a combination therapy is used, the compound of the present invention and the combined therapeutic agent may be present in the same or different pharmaceutical compositions, and may be administered separately, sequentially or simultaneously.
另一方面，本发明的化合物可以联合视网膜的激光治疗给药。激光光凝联合抗VEGF药物(如雷珠单抗)治疗糖尿病黄斑水肿为本领域所熟知且被验证疗效显著。On the other hand, the compound of the present invention can be administered in combination with laser treatment of the retina. Laser photocoagulation combined with anti-VEGF drugs (such as ranibizumab) in the treatment of diabetic macular edema is well known in the art and has been proven to have significant efficacy.
定义definition
如本文所用，术语“烷基”自身或作为另一取代基的部分意指(除非另外说明)具有指定碳原子数目的饱和直链或支链烃基。包括：至多10个碳原子(C 1-C 10)，或至多6个碳原子(C 1-C 6)，或至多4个碳原子(C 1-C 4)，或至多3个碳原子(C 1-C 3)的直链基团；或者3至10个碳原子(C 3-C 10)，或至多7个碳原子(C 3-C 7)，或至多4个碳原子(C 3-C 4)的支链基团。烷基的实例包括但不限于甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基、仲丁基、正戊基、正己基、正庚基、正辛基。 As used herein, the term "alkyl" by itself or as part of another substituent means (unless otherwise stated) a saturated linear or branched hydrocarbon group having the specified number of carbon atoms. Including: up to 10 carbon atoms (C 1 -C 10 ), or up to 6 carbon atoms (C 1 -C 6 ), or up to 4 carbon atoms (C 1 -C 4 ), or up to 3 carbon atoms ( C 1 -C 3 ) linear group; or 3 to 10 carbon atoms (C 3 -C 10 ), or up to 7 carbon atoms (C 3 -C 7 ), or up to 4 carbon atoms (C 3 -C 4 ) branched chain group. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n- Sinki.
如本文所用，“烷氧基”代表通过氧桥连接的具有特定数目碳原子的上述烷基。包括：1 至6个碳原子(C 1-C 6)，或1至4个碳原子(C 1-C 4)的直链基团；-3至6个碳原子(C 3-C 6)，或3至4个碳原子(C 3-C 4)的支链基团。烷氧基的实例包括但不限于：甲氧基、乙氧基、正丙氧基、异丙氧基、正丁氧基、仲丁氧基、叔丁氧基、正戊氧基和S-戊氧基。 As used herein, "alkoxy" represents the aforementioned alkyl group having a specified number of carbon atoms connected by an oxygen bridge. Including: 1 to 6 carbon atoms (C 1 -C 6 ), or 1 to 4 carbon atoms (C 1 -C 4 ) linear groups; -3 to 6 carbon atoms (C 3 -C 6 ) , Or a branched group with 3 to 4 carbon atoms (C 3 -C 4 ). Examples of alkoxy groups include, but are not limited to: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy and S- Pentoxy.
如本文所用，“环烷基”是3至7个碳原子的单环饱和烃基。环烷基可以含有3至7个碳原子或3至6个碳原子或3至5个碳原子或3至4个碳原子。单环环烷基的实例包括环丙基、环丁基、环戊基、环己基和环庚基。As used herein, "cycloalkyl" is a monocyclic saturated hydrocarbon group of 3 to 7 carbon atoms. The cycloalkyl group may contain 3 to 7 carbon atoms or 3 to 6 carbon atoms or 3 to 5 carbon atoms or 3 to 4 carbon atoms. Examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.
如本文所用，“卤代烷基”是如上述的烷基上氢原子部分或全部被卤素原子取代形成的基团；除非另有说明，卤素选自Cl、F、Br和I。As used herein, "haloalkyl" is a group formed by partially or completely replacing hydrogen atoms on an alkyl group with halogen atoms as described above; unless otherwise specified, halogen is selected from Cl, F, Br, and I.
如本文所用，术语“芳环”是指单环、双环或多环芳烃基团，例如苯、萘。术语“芳基”还包括具有两个或多个环系，其中两个或多个碳为两个相邻环共用(所述环为“稠合环”)，其中至少一个环为芳族烃，其他的环可以是如环烷基、环烯基、环炔基、芳基、杂芳基和/或杂环基。在某些实施例中，术语“芳基”是指苯基基团。在某些实施例中，“芳基”具有6至10个碳原子。As used herein, the term "aromatic ring" refers to a monocyclic, bicyclic or polycyclic aromatic hydrocarbon group, such as benzene, naphthalene. The term "aryl" also includes two or more ring systems in which two or more carbons are shared by two adjacent rings (the ring is a "fused ring"), and at least one ring is an aromatic hydrocarbon , Other rings can be, for example, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and/or heterocyclic group. In certain embodiments, the term "aryl" refers to a phenyl group. In certain embodiments, "aryl" has 6 to 10 carbon atoms.
如本文所用，术语“芳杂环”是指单环、双环和多环芳族基团，在环结构中具有3至12个总原子，其中包括一个或多个杂原子，如氮、氧或硫。示例性芳杂环团包括氮杂吲哚基、苯并(b)噻吩基、苯并咪唑基、苯并呋喃基、苯并恶唑基、苯并噻唑基、苯并噻二唑基、苯并三唑基、苯并恶二唑基、呋喃基、咪唑基、咪唑并吡啶基、吲哚基、吲哚啉基、吲唑基、异吲哚啉基、异恶唑基、异噻唑基、异喹啉基、恶唑基、嘌呤基、吡喃基、吡嗪基、吡唑基、吡啶基、嘧啶基、吡咯基、吡咯并[2,3-d]嘧啶基、吡唑并[3,4-d]嘧啶基、喹啉基、喹唑啉基、三唑基、噻唑基、噻吩基、四氢吲哚基、四唑基、噻二唑基、噻吩基、硫代吗啉基、三唑基或托烷基等等。术语“芳杂环”还包括具有两个或多个环的多环系，其中两个或多个碳为两个相邻环共用(所述环是“稠合环”)，其中至少一个环是在环结构中具有一个或多个杂原子的芳族基团，例如，其他的环可以是环烷基、环烯基、环炔基、芳基、杂芳基和/或杂环基。As used herein, the term "aromatic heterocyclic ring" refers to monocyclic, bicyclic and polycyclic aromatic groups having 3 to 12 total atoms in the ring structure, including one or more heteroatoms such as nitrogen, oxygen or sulfur. Exemplary aromatic heterocyclic groups include azaindolyl, benzo(b)thienyl, benzimidazolyl, benzofuranyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzene O-triazolyl, benzoxadiazolyl, furyl, imidazolyl, imidazopyridyl, indolyl, indolinyl, indazolyl, isoindolinyl, isoxazolyl, isothiazolyl , Isoquinolinyl, oxazolyl, purinyl, pyranyl, pyrazinyl, pyrazolyl, pyridinyl, pyrimidinyl, pyrrolyl, pyrrolo[2,3-d]pyrimidinyl, pyrazolo[ 3,4-d]pyrimidinyl, quinolinyl, quinazolinyl, triazolyl, thiazolyl, thienyl, tetrahydroindolyl, tetrazolyl, thiadiazolyl, thienyl, thiomorpholine Group, triazolyl or tropyl group and so on. The term "aromatic heterocyclic ring" also includes a polycyclic ring system having two or more rings, in which two or more carbons are shared by two adjacent rings (the ring is a "fused ring"), in which at least one ring It is an aromatic group having one or more heteroatoms in the ring structure, for example, other rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and/or heterocyclic group.
如本文所用，“芳杂双环”指双环芳族基团，在环结构中具有3至12个总原子，其中包括一个或多个杂原子，如氮、氧或硫。示例性芳杂双环包括氮杂吲哚基、苯并(b)噻吩基、苯并咪唑基、苯并呋喃基、苯并恶唑基、苯并噻唑基、苯并噻二唑基、苯并三唑基、苯并恶二唑基、咪唑并吡啶基、吲哚基、吲哚啉基、吲唑基、异吲哚啉基、异恶唑基、异噻唑基、异喹啉基、吡咯并[2,3-d]嘧啶基、吡咯并[2,3-b]吡啶、吡唑并[3,4-d]嘧啶基、喹啉基、喹唑啉等等。术语“芳杂环”还包括具有两个或多个环的多环系，其中两个或多个碳为两个相邻环共用(所述环是“稠合环”)，其中至少一个环是在环结构中具有一个或多个杂原子的芳族基团，例如，其他的环可以是环烷基、环烯基、环炔基、芳基、杂芳基和/或杂环基。As used herein, "aromatic heterobicyclic ring" refers to a bicyclic aromatic group having 3 to 12 total atoms in the ring structure, including one or more heteroatoms such as nitrogen, oxygen, or sulfur. Exemplary heterobicyclic rings include azaindolyl, benzo(b)thienyl, benzimidazolyl, benzofuranyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzo Triazolyl, benzoxadiazolyl, imidazopyridyl, indolyl, indolinyl, indazolyl, isoindolinyl, isoxazolyl, isothiazolyl, isoquinolinyl, pyrrole And [2,3-d]pyrimidinyl, pyrrolo[2,3-b]pyridine, pyrazolo[3,4-d]pyrimidinyl, quinolinyl, quinazoline, etc. The term "aromatic heterocyclic ring" also includes a polycyclic ring system having two or more rings, in which two or more carbons are shared by two adjacent rings (the ring is a "fused ring"), in which at least one ring It is an aromatic group having one or more heteroatoms in the ring structure, for example, other rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and/or heterocyclic group.
如本文所用，术语“单环或稠合多环非芳族环系”是指排除芳族或杂芳族以外的所有环系，包括饱和或不饱和的单环或稠合多环(包括双环)，其可以任选包含0-3个独立选自N、O和S的杂原子，含杂原子的基团不具有芳香性，在环结构中具有5-10个总原子。所述“单环或稠合多环非芳族环系”可以包括但不限于：5元单环，如四氢呋喃基、四氢噻吩基、二氧杂环戊烯基、吡咯烷基、咪唑烷基、吡唑烷基、恶唑烷基，吡咯啉基；或6元单环，如四氢吡喃基、哌啶基、吗啉基、二噻烷基、硫代吗啉基、哌嗪基或三噻烷基；或7元但环，如二氮杂环庚烷基。任选地，所述“单环或稠合多环非芳族环系”可以是苯并稠合的，例如但不限于二氢异喹啉基。所述“单环或稠合多环非芳族环系”可以是双环的，例如但不限于氮杂双环[3.1.0]己烷，六氢环戊并[c]吡咯-2(1H)-基环，六氢吡咯并[1,2-a]吡嗪-2(1H)-基环。As used herein, the term "monocyclic or fused polycyclic non-aromatic ring system" refers to excluding all ring systems other than aromatic or heteroaromatic, including saturated or unsaturated monocyclic or fused polycyclic (including bicyclic ), which may optionally contain 0-3 heteroatoms independently selected from N, O, and S. The heteroatom-containing group does not have aromaticity and has 5-10 total atoms in the ring structure. The "monocyclic or fused polycyclic non-aromatic ring system" may include but is not limited to: 5-membered monocyclic ring, such as tetrahydrofuryl, tetrahydrothienyl, dioxolyl, pyrrolidinyl, imidazolidine Group, pyrazolidinyl, oxazolidinyl, pyrrolinyl; or 6-membered monocyclic ring, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithiaalkyl, thiomorpholinyl, piperazine Group or trithiaalkyl; or 7-membered but ring, such as diazeppanyl. Optionally, the "monocyclic or fused polycyclic non-aromatic ring system" may be benzo-fused, such as but not limited to dihydroisoquinolinyl. The "monocyclic or fused polycyclic non-aromatic ring system" may be bicyclic, such as but not limited to azabicyclo[3.1.0]hexane, hexahydrocyclopenta[c]pyrrole-2(1H) -Yl ring, hexahydropyrrolo[1,2-a]pyrazine-2(1H)-yl ring.
如本文所用，术语“被取代的”是指特定原子上的任意一个或多个氢原子被取代基取代，可以包括重氢和氢的变体，只要特定原子的价态是正常的并且取代后的化合物是稳定的。当取代基为羰基(即＝O)时，意味着两个氢原子被取代。羰基取代不会发生在芳香基上。术语“任选地被下述取代基取代”是指可以被取代，也可以不被取代，除非另有规定，取代基的种类和数目在化学上可以实现的基础上可以是任意的。As used herein, the term "substituted" means that any one or more hydrogen atoms on a specific atom is replaced by a substituent, and may include deuterium and hydrogen variants, as long as the valence of the specific atom is normal and after substitution The compound is stable. When the substituent is a carbonyl group (ie, =0), it means that two hydrogen atoms are replaced. Carbonyl substitution does not occur on aromatic groups. The term "optionally substituted by the following substituents" means that it may be substituted or unsubstituted. Unless otherwise specified, the type and number of the substituents may be arbitrary on the basis that they can be chemically realized.
如本文所用，在化学结构中与键相交的波浪线指示键的附连点，其在化学结构中的波浪键与分子的其余部分相交。As used herein, the wavy line that intersects the bond in the chemical structure indicates the attachment point of the bond, and its wavy bond in the chemical structure intersects the rest of the molecule.
如本文所用，“药学上可接受的盐”，包括衍生自无机酸或有机酸的盐，所述无机酸或有机酸包括但不限于盐酸、氢溴酸、硫酸、硝酸、高氯酸、磷酸、甲酸、乙酸、乳酸、马来酸、富马酸、琥珀酸、酒石酸、乙醇酸、水杨酸、柠檬酸、甲磺酸、苯磺酸、苯甲酸、丙二酸、三氟乙酸、三氯乙酸、萘-2-磺酸和其他酸。药学上可接受的盐包括化合物碱与盐的分子摩尔比例不是1:1的情形。作为一个实例，盐可以包含相对于每个碱分子有两个分子的有机酸或者无机酸。作为另一个实例，盐可以包含相对于每个碱分子有1/2个分子的有机酸或者无机酸。As used herein, "pharmaceutically acceptable salts" include salts derived from inorganic or organic acids including but not limited to hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, phosphoric acid , Formic acid, acetic acid, lactic acid, maleic acid, fumaric acid, succinic acid, tartaric acid, glycolic acid, salicylic acid, citric acid, methanesulfonic acid, benzenesulfonic acid, benzoic acid, malonic acid, trifluoroacetic acid, three Chloroacetic acid, naphthalene-2-sulfonic acid and other acids. The pharmaceutically acceptable salt includes the case where the molar ratio of the compound base to the salt is not 1:1. As an example, the salt may contain two molecules of organic acid or inorganic acid per base molecule. As another example, the salt may contain 1/2 molecule of organic acid or inorganic acid per base molecule.
如本文所用，“溶剂化物”是指本申请化合物与一个或多个溶剂分子的物理缔合；该物理缔合涉及各种程度的离子键和共价键，其包括氢键；在某些情况下，例如当将一个或多个溶剂分子引入到结晶固体的晶格中时，溶剂化物将能够被分离；“溶剂化物”涵盖溶液相和可分离的溶剂化物；合适的溶剂非限制性实例包括但不限于异丙醇、乙醇、甲醇、DMSO、乙酸乙酯、乙酸和乙醇胺等；“水合物”为其中溶剂分子为H 2O的溶剂化物。 As used herein, "solvate" refers to the physical association of a compound of the present application with one or more solvent molecules; this physical association involves various degrees of ionic and covalent bonds, including hydrogen bonds; in some cases Below, for example, when one or more solvent molecules are introduced into the crystal lattice of a crystalline solid, the solvate will be able to be separated; “solvate” encompasses both solution phase and isolatable solvate; non-limiting examples of suitable solvents include But not limited to isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, ethanolamine, etc.; "hydrate" is a solvate in which the solvent molecule is H 2 O.
除非另有规定，本文所描绘的结构也意味着包括该结构的所有异构(例如对映异构、非对映异构和几何异构(或构象异构))形式；例如每一不对称中心的R与S构型，(Z)与(E)双键异构体，和(Z)与(E)构象异构体。因此，这些化合物的单一立体化学异构体以及对映异构、非对映异构和几何异构(或构象异构)混合物都属于本发明的范围。除非另有规定，本发明化合物的所有互变异构形式都属于本发明的范围。Unless otherwise specified, the structure described herein is also meant to include all isomeric (such as enantiomers, diastereomers, and geometric (or conformational)) forms of the structure; for example, each asymmetry The R and S configurations of the center, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers. Therefore, single stereochemical isomers and enantiomeric, diastereomeric, and geometric (or conformational) mixtures of these compounds fall within the scope of the present invention. Unless otherwise specified, all tautomeric forms of the compounds of the present invention belong to the scope of the present invention.
本发明包括所有可药用的同位素标记的化合物，即式(I)化合物中一或多个原子被具有相同原子数但通常与自然形成的原子质量或质量数不同的原子替代。本发明化合物中适合的同位素实例包括但不限于：氢的同位素，例如 2H和 3H；碳的同位素，例如 11C、 13C和 14C；氯的同位素，例如 36Cl；氟的同位素，例如 18F；碘的同位素，例如 123I和 125I；氮的同位素，例如 13N和 15N；氧的同位素，例如 15O、 17O和 18O；磷的同位素，例如 32P；和硫的同位素，例如 35S。 The present invention includes all pharmaceutically acceptable isotopically-labeled compounds, that is, one or more atoms in the compound of formula (I) are replaced by atoms having the same atomic number but usually different from the naturally-occurring atomic mass or mass number. Examples of suitable isotopes in the compounds of the present invention include, but are not limited to: isotopes of hydrogen, such as 2 H and 3 H; isotopes of carbon, such as 11 C, 13 C, and 14 C; isotopes of chlorine, such as 36 Cl; isotopes of fluorine, For example 18 F; isotopes of iodine, such as 123 I and 125 I; isotopes of nitrogen, such as 13 N and 15 N; isotopes of oxygen, such as 15 O, 17 O, and 18 O; isotopes of phosphorus, such as 32 P; and sulfur Isotope of, for example 35 S.
如本文所用，“前药”是指在体内通过代谢方式(例如，通过水解、还原或氧化)而可转化成本发明化合物的化合物。As used herein, "prodrug" refers to a compound that can be converted into a compound of the invention in vivo by means of metabolism (for example, by hydrolysis, reduction, or oxidation).
术语“药学上可接受的载体”是指能够递送本发明有效量活性化合物、不干扰活性化合物生物活性并且对宿主或者患者无毒副作用的任何制剂或载体介质，包括任何本领域普通技术人员已知的溶剂、分散介质、包衣材料、表面活性剂、抗氧剂、防腐剂(例如抗菌剂、抗真菌剂)、等张剂、吸收延迟剂、盐、防腐剂、药物、药物稳定剂、粘合剂、赋形剂、崩解剂、润滑剂、甜味剂、矫味剂、染料等及其组合The term "pharmaceutically acceptable carrier" refers to any preparation or carrier medium that can deliver an effective amount of the active compound of the present invention, does not interfere with the biological activity of the active compound, and has no toxic or side effects to the host or patient, including any preparation or carrier medium known to those of ordinary skill in the art Solvents, dispersion media, coating materials, surfactants, antioxidants, preservatives (such as antibacterial agents, antifungal agents), isotonic agents, absorption delaying agents, salts, preservatives, drugs, drug stabilizers, viscosity Mixtures, excipients, disintegrants, lubricants, sweeteners, correctives, dyes, etc. and combinations thereof
术语“赋形剂”在本文中用来描述不同于本发明的化合物的任何成分，其可以对所述制剂赋予功能性(如药物释放速率控制)和/或非功能性(如加工助剂或稀释剂)。赋形剂的选择很大 程度上取决于诸如具体给药方式、赋形剂对溶解度和稳定性的影响、以及剂型的性质等因素。示例性的赋形剂可以包括但不限于缓冲剂、稳定剂、表面活性剂、湿润剂、润滑剂、乳化剂、悬浮剂、防腐剂、抗氧化剂、不透明剂、助流剂、加工助剂、着色剂、增甜剂、芳香剂、调味剂、稀释剂和其它已知的添加剂中的一种或多种。The term "excipient" is used herein to describe any ingredient other than the compound of the present invention, which can impart functionality (such as drug release rate control) and/or non-functionality (such as processing aids or Thinner). The choice of excipients largely depends on factors such as the specific mode of administration, the effect of the excipients on solubility and stability, and the nature of the dosage form. Exemplary excipients may include, but are not limited to, buffers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacity agents, glidants, processing aids, One or more of colorants, sweeteners, fragrances, flavors, diluents and other known additives.
本发明化合物可以以任何便利的使用形式被施用，如片剂、粉末、胶囊、溶液、分散体、悬浮液、糖浆剂、喷雾剂、栓剂、凝胶剂、乳剂、贴剂等。此类组合物可以包含药物制剂中常规的组分，如稀释剂、载体、pH调节剂、增甜剂、填充剂和另外的活性剂。本发明的活性化合物也可以被配制成持续释放的剂型。The compounds of the present invention can be administered in any convenient use form, such as tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches and the like. Such compositions may contain conventional components in pharmaceutical preparations, such as diluents, carriers, pH adjusters, sweeteners, fillers, and additional active agents. The active compounds of the present invention can also be formulated into sustained-release dosage forms.
本发明化合物可以通过任何合适的方式施用，包括口服、局部(包括颊部和舌下)、直肠、阴道、透皮、肠胃外、皮下、腹膜内、肺内、皮肤内、鞘内和硬膜外以及鼻内，并且，如果需要用于局部治疗，病灶内施用。肠胃外输注包括肌肉内、静脉内、动脉内、腹膜内、大脑内、眼内、病灶内或皮下施用。The compounds of the present invention can be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal and dural Externally and intranasally, and, if necessary, for topical treatment, intralesional administration. Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal, intracerebral, intraocular, intralesional, or subcutaneous administration.
对于病症如与糖尿病视网膜病变和糖尿病黄斑水肿相关的视网膜血管通透性疾病的治疗，本发明的化合物可以以适于注射到患者眼内部区域的形式，尤其是，以适于玻璃体内注射的形式施用。For the treatment of disorders such as retinal vascular permeability diseases related to diabetic retinopathy and diabetic macular edema, the compound of the present invention may be in a form suitable for injection into the intraocular region of a patient, especially, in a form suitable for intravitreal injection Apply.
本发明化合物术语“治疗有效的量”指引起患者生物学或医学反应的本发明化合物的量，所述反应为例如降低或抑制酶或蛋白活性或者改善症状、缓解病症、减慢或延迟疾病发展或者预防疾病等。在一个非限制性实施方案中，术语“治疗有效的量”指当给药于细胞或组织或非细胞性生物学物质或介质时，本发明化合物的量能够至少部分降低或抑制血浆激肽释放酶活性；或至少部分降低或抑制血浆激肽释放酶表达。化合物、药用组合物或结合的治疗有效剂量取决于患者的种属、体重、年龄和个体情况、疾病或病症或者需要治疗的严重性。本发明化合物和组合物总的每日用量将由主治医师在合理的医学判断范围内决定。The term "therapeutically effective amount" of the compound of the present invention refers to the amount of the compound of the present invention that causes a patient's biological or medical response, such as reducing or inhibiting enzyme or protein activity or improving symptoms, alleviating symptoms, slowing down or delaying disease progression Or prevent disease, etc. In a non-limiting embodiment, the term "therapeutically effective amount" refers to the amount of the compound of the present invention that is capable of at least partially reducing or inhibiting plasma kallikrein when administered to cells or tissues or non-cellular biological substances or mediators. Enzyme activity; or at least partially reduce or inhibit plasma kallikrein expression. The therapeutically effective dose of the compound, pharmaceutical composition or combination depends on the patient's species, weight, age and individual condition, disease or condition, or the severity of the need for treatment. The total daily dosage of the compounds and compositions of the present invention will be determined by the attending physician within the scope of reasonable medical judgment.
化合物经手工或者Chemdraw软件命名，市售化合物采用供应商目录名称。The compounds are named manually or by Chemdraw software, and the commercially available compounds use the supplier catalog name.
与现有技术相比，本发明的主要优点在于：Compared with the prior art, the main advantages of the present invention are:
提供了一系列结构新颖、活性好、选择性高的的三环类血浆激肽释放酶抑制剂化合物，可广泛用于涉及血浆激肽释放酶活性相关的疾病的预防或治疗。A series of tricyclic plasma kallikrein inhibitor compounds with novel structure, good activity and high selectivity are provided, which can be widely used in the prevention or treatment of diseases related to plasma kallikrein activity.
附图说明Description of the drawings
图1显示化合物22、6、71和73化合物对大鼠中CA-1刺激的血管通透性模型中的荧光素渗漏评分；Figure 1 shows the fluorescein leakage scores of compounds 22, 6, 71 and 73 in a CA-1 stimulated vascular permeability model in rats;
图2显示化合物1、6、22和73化合物对大鼠中CA-1刺激的血管通透性模型中的视网膜厚度变化；Figure 2 shows the changes in retinal thickness of compounds 1, 6, 22 and 73 in a CA-1 stimulated vascular permeability model in rats;
图3显示部分化合物对视网膜色素上皮细胞ARPE19的毒性。Figure 3 shows the toxicity of some compounds to ARPE19 of retinal pigment epithelial cells.
具体实施方式Detailed ways
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，通常按照常规条件或按照制造厂商所建议的条件。实施例(制备)中描述的化学反应可按照本领域熟知的方法制备本发明的其它化合物，并且用于制备本发明化合物的其他方法属于本发明的范围。The present invention will be further explained below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods that do not indicate specific conditions in the following examples are usually in accordance with conventional conditions or in accordance with the conditions recommended by the manufacturer. The chemical reactions described in the examples (preparation) can be used to prepare other compounds of the present invention according to methods well known in the art, and other methods for preparing the compounds of the present invention fall within the scope of the present invention.
实施例1Example 1
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000009
-2-甲酰胺(化合物1)的制备 N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11 -Dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000009
Preparation of -2-carboxamide (Compound 1)
Figure PCTCN2020137640-appb-000010
Figure PCTCN2020137640-appb-000010
步骤a)：2-氨基-2-(羟基亚氨基)乙酸乙酯的制备Step a): Preparation of ethyl 2-amino-2-(hydroxyimino)acetate
将水(110mL)缓慢加入氰基甲酸乙酯(30.0g，0.303mol)、盐酸羟胺(31.6g，0.455mol)和碳酸钠(80.3g，0.758mol)的乙醇(200mL)混合液中，加毕，反应液20℃搅拌反应10h，反应结束后减压蒸除溶剂，残余物以乙酸乙酯(200mL×3)萃取，合并有机层，无水硫酸钠干燥，过滤，滤液减压蒸干，得2-氨基-2-(羟基亚氨基)乙酸乙酯，收率65.0％， 1H NMR(400MHz,DMSO-d 6)δ10.66-9.12(m,1H),8.24(s,2H),5.79-5.31(m,2H),4.07-3.97(m,2H),1.18-1.13(m,3H)，ESI-MS(m/z)：133.2[M+H] +。 Slowly add water (110mL) to ethyl cyanoformate (30.0g, 0.303mol), hydroxylamine hydrochloride (31.6g, 0.455mol) and sodium carbonate (80.3g, 0.758mol) in ethanol (200mL) mixture, and the addition is complete. The reaction solution was stirred at 20°C for 10 hours. After the reaction, the solvent was evaporated under reduced pressure. The residue was extracted with ethyl acetate (200mL×3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated under reduced pressure to obtain 2-Amino-2-(hydroxyimino) ethyl acetate, yield 65.0%, 1 H NMR (400MHz, DMSO-d 6 ) δ 10.66-9.12 (m, 1H), 8.24 (s, 2H), 5.79 -5.31 (m, 2H), 4.07-3.97 (m, 2H), 1.18-1.13 (m, 3H), ESI-MS (m/z): 133.2 [M+H] + .
步骤b)：3-(((1-氨基-2-乙氧基-2-氧亚乙基)氨基)氧基)丙烯酸叔丁酯的制备Step b): Preparation of 3-(((1-amino-2-ethoxy-2-oxyethylene)amino)oxy) tert-butyl acrylate
将2-氨基-2-(羟基亚氨基)乙酸乙酯(9.0g，68.120mmol)、丙炔酸叔丁酯(8.59g，68.120mmol)、三乙胺(10.4mL，74.932mmol)和乙醇(90mL)加入反应瓶中，35℃搅拌反应 10h，反应结束后，减压蒸除溶剂，残余物加水(150mL)稀释，用乙酸乙酯(150mL×3)萃取，合并有机层，饱和氯化钠水溶液洗涤，无水硫酸钠干燥，过滤，滤液减压蒸干，所得粗品经硅胶柱层析纯化(洗脱剂：石油醚/乙酸乙酯＝3/1)，得3-(((1-氨基-2-乙氧基-2-氧亚乙基)氨基)氧基)丙烯酸叔丁酯，收率62.5％， 1H NMR(400MHz,CDCl 3)δ7.85(d,J＝12.4Hz,1H),5.56(d,J＝12.4Hz,1H),5.37(br s,2H),4.39(q,J＝7.2Hz,2H),1.47(s,9H),1.42-1.37(m,3H)，ESI-MS(m/z)：297.2[M+K] +。 Combine 2-amino-2-(hydroxyimino) ethyl acetate (9.0g, 68.120mmol), tert-butyl propiolate (8.59g, 68.120mmol), triethylamine (10.4mL, 74.932mmol) and ethanol ( 90mL) was added to the reaction flask, and the reaction was stirred at 35°C for 10h. After the reaction, the solvent was evaporated under reduced pressure. The residue was diluted with water (150mL) and extracted with ethyl acetate (150mL×3). The organic layers were combined and saturated sodium chloride. Wash with aqueous solution, dry with anhydrous sodium sulfate, filter, and evaporate the filtrate under reduced pressure. The crude product obtained is purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate = 3/1) to give 3-(((1- Amino-2-ethoxy-2-oxyethylene)amino)oxy) tert-butyl acrylate, the yield is 62.5%, 1 H NMR (400MHz, CDCl 3 ) δ 7.85 (d, J = 12.4 Hz, 1H), 5.56 (d, J = 12.4 Hz, 1H), 5.37 (br s, 2H), 4.39 (q, J = 7.2 Hz, 2H), 1.47 (s, 9H), 1.42-1.37 (m, 3H) , ESI-MS (m/z): 297.2 [M+K] + .
步骤c)：1H-咪唑-2-甲酸乙酯-4-甲酸叔丁酯的制备Step c): Preparation of 1H-imidazole-2-carboxylic acid ethyl ester-4-carboxylic acid tert-butyl ester
将3-(((1-氨基-2-乙氧基-2-氧亚乙基)氨基)氧基)丙烯酸叔丁酯(2.0g，7.744mmol)和二甲苯(15mL)加入反应瓶中，155℃微波反应3h，反应结束后，减压蒸除溶剂，残余物经硅胶柱层析纯化(洗脱剂：石油醚/乙酸乙酯＝3/1)，得黄色固体，收率89.2％， 1H NMR(400MHz,CDCl 3)δ7.85-7.57(m,1H),4.41-4.32(m,2H),1.51(d,J＝2.4Hz,9H),1.38-1.32(m,3H)。 Add 3-(((1-amino-2-ethoxy-2-oxyethylene)amino)oxy) tert-butyl acrylate (2.0g, 7.744mmol) and xylene (15mL) into the reaction flask, Microwave reaction at 155°C for 3 hours. After the reaction, the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate = 3/1) to obtain a yellow solid with a yield of 89.2%. 1 H NMR (400MHz, CDCl 3 ) δ 7.85-7.57 (m, 1H), 4.41-4.32 (m, 2H), 1.51 (d, J=2.4 Hz, 9H), 1.38-1.32 (m, 3H).
步骤d)：4-甲基-2-硝基-苯甲酰氯的制备Step d): Preparation of 4-methyl-2-nitro-benzoyl chloride
将草酰氯(200mL)在15℃条件下滴加至4-甲基-2-硝基-苯甲酸(43.0g，0.237mol)和二氯甲烷(200mL)的混合液中，滴毕，升温至45℃反应5h，反应结束后，减压浓缩，得4-甲基-2-硝基-苯甲酰氯，无需进一步纯化直接用于下一步反应。Add oxalyl chloride (200mL) dropwise to the mixture of 4-methyl-2-nitro-benzoic acid (43.0g, 0.237mol) and dichloromethane (200mL) at 15°C. The reaction was conducted at 45°C for 5 hours. After the reaction was completed, it was concentrated under reduced pressure to obtain 4-methyl-2-nitro-benzoyl chloride, which was directly used in the next reaction without further purification.
步骤e)：4-甲基-2-硝基-苯甲酸甲酯的制备Step e): Preparation of 4-methyl-2-nitro-benzoic acid methyl ester
将4-甲基-2-硝基-苯甲酰氯(50.0g，0.248mol)的二氯甲烷(100mL)溶液在15℃缓慢滴入甲醇(200mL)和三乙胺(51.7mL，0.372mol)的混合液中，滴毕，15℃搅拌反应5h，反应结束后，减压蒸除溶剂，残余物加水(500mL)稀释，用乙酸乙酯萃取(600mL×3)，合并有机层，饱和氯化钠水溶液(600mL×2)洗涤，无水硫酸钠干燥，过滤，滤液减压蒸干，得黄色固体，所得粗品直接用于下一步反应，粗品收率82.6％， 1H NMR(400MHz,CDCl 3)δ7.69-7.56(m,2H),7.44(br d,J＝8.0Hz,1H),3.91-3.85(m,3H),2.56-2.27(m,3H)。 A solution of 4-methyl-2-nitro-benzoyl chloride (50.0g, 0.248mol) in dichloromethane (100mL) was slowly dropped into methanol (200mL) and triethylamine (51.7mL, 0.372mol) at 15°C After dripping, stir the reaction at 15°C for 5h. After the reaction, the solvent was evaporated under reduced pressure. The residue was diluted with water (500mL) and extracted with ethyl acetate (600mL×3). The organic layers were combined and saturated chlorinated. Washed with sodium aqueous solution (600mL×2), dried with anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain a yellow solid. The obtained crude product was directly used in the next reaction. The crude product yield was 82.6%. 1 H NMR (400MHz, CDCl 3 ) δ 7.69-7.56 (m, 2H), 7.44 (br d, J=8.0 Hz, 1H), 3.91-3.85 (m, 3H), 2.56-2.27 (m, 3H).
步骤f)：4-溴甲基-2-硝基-苯甲酸甲酯的制备Step f): Preparation of 4-bromomethyl-2-nitro-benzoic acid methyl ester
将4-甲基-2-硝基-苯甲酸甲酯(40.0g，0.204mol)的乙腈(200mL)溶液在0℃滴入N-溴代琥珀酰亚胺(NBS，36.4g，0.204mol)、过氧化二苯甲酰(BPO，2.98g，12.302mmol)和乙腈(100mL)的混合液中，滴毕，在氮气保护下升温至80℃反应4h，反应结束后，减压蒸除溶剂，残余物加水(400mL)稀释，用乙酸乙酯萃取(500mL×3)，合并有机层，饱和氯化钠水溶液(300mL×2)洗涤，无水硫酸钠干燥，过滤，滤液减压蒸干，得4-溴甲基-2-硝基-苯甲酸甲酯，粗品直接用于下一步反应。A solution of 4-methyl-2-nitro-benzoic acid methyl ester (40.0g, 0.204mol) in acetonitrile (200mL) was dropped into N-bromosuccinimide (NBS, 36.4g, 0.204mol) at 0°C , Dibenzoyl peroxide (BPO, 2.98g, 12.302mmol) and acetonitrile (100mL) mixed solution, dripped, heated to 80 ℃ under the protection of nitrogen to react for 4h, after the completion of the reaction, the solvent was evaporated under reduced pressure, The residue was diluted with water (400 mL), extracted with ethyl acetate (500 mL×3), the organic layers were combined, washed with saturated aqueous sodium chloride solution (300 mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure. 4-bromomethyl-2-nitro-benzoic acid methyl ester, the crude product was directly used in the next reaction.
步骤g)：4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-2-硝基苯甲酸甲酯的制备Step g): Preparation of methyl 4-((3-bromo-5-methyl-1H-pyrazol-1-yl)methyl)-2-nitrobenzoate
将4-溴甲基-2-硝基-苯甲酸甲酯(24.0g，87.578mmol)、3-溴-5-甲基-1H-吡唑(14.1g，87.578mmol)、碳酸钾(24.2g，175.155mmol)和乙腈(400mL)加入反应瓶中，40℃搅拌反应50h，反应结束后减压蒸除溶剂，残余物加水(200mL)稀释，用乙酸乙酯(200mL×3)萃取，合并有机层，饱和氯化钠水溶液(100mL×3)洗涤，无水硫酸钠干燥，过滤，滤液减压蒸干，所得粗品经硅胶柱层析纯化(洗脱剂：石油醚/乙酸乙酯＝3/1)，得4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-2-硝基苯甲酸甲酯，收率50.9％， 1H NMR(400MHz,CDCl 3)δ7.70(d,J＝8.0Hz,1H),7.61(d,J＝1.6Hz,1H),7.40-7.34(m,1H),6.12(d,J＝0.8Hz,1H),5.32(s,2H),3.93-3.86(m,3H),2.21(d,J＝0.8Hz,3H)；ESI-MS(m/z)：354.1[M+H] +。 Combine 4-bromomethyl-2-nitro-benzoic acid methyl ester (24.0g, 87.578mmol), 3-bromo-5-methyl-1H-pyrazole (14.1g, 87.578mmol), potassium carbonate (24.2g , 175.155mmol) and acetonitrile (400mL) were added to the reaction flask, stirred at 40°C for 50h. After the reaction, the solvent was evaporated under reduced pressure. The residue was diluted with water (200mL), extracted with ethyl acetate (200mL×3), and combined organic The layer was washed with saturated sodium chloride aqueous solution (100mL×3), dried with anhydrous sodium sulfate, filtered, the filtrate was evaporated to dryness under reduced pressure, and the crude product obtained was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate = 3/ 1) to obtain methyl 4-((3-bromo-5-methyl-1H-pyrazol-1-yl)methyl)-2-nitrobenzoate with a yield of 50.9%. 1 H NMR (400MHz, CDCl 3 )δ7.70(d,J=8.0Hz,1H), 7.61(d,J=1.6Hz,1H), 7.40-7.34(m,1H), 6.12(d,J=0.8Hz,1H), 5.32 (s, 2H), 3.93-3.86 (m, 3H), 2.21 (d, J = 0.8 Hz, 3H); ESI-MS (m/z): 354.1 [M+H] + .
步骤h)：(4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-2-硝基苯甲醇的制备Step h): Preparation of (4-((3-bromo-5-methyl-1H-pyrazol-1-yl)methyl)-2-nitrobenzyl alcohol
在0℃将硼氢化钠(2.32g，61.326mmol)和氯化钙(6.82g，61.441mmol)加入4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-2-硝基苯甲酸甲酯(14.5g，40.942mmol)的四氢呋喃(200mL)溶液中，0℃搅拌反应5h，反应液在0℃用氯化铵水溶液淬灭，用乙酸乙酯(200mL×3)萃取，合并有机层，饱和氯化钠水溶液(100mL×3)洗涤，无水硫酸钠干燥，过滤，滤液减压蒸干，得(4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-2-硝基苯甲醇，无需纯化直接用于下一步反应，ESI-MS(m/z)：326.0[M+H] +。 Add sodium borohydride (2.32g, 61.326mmol) and calcium chloride (6.82g, 61.441mmol) to 4-((3-bromo-5-methyl-1H-pyrazol-1-yl)methyl at 0°C )-2-nitrobenzoic acid methyl ester (14.5g, 40.942mmol) in tetrahydrofuran (200mL), stirred at 0°C for 5h, the reaction solution was quenched with aqueous ammonium chloride solution at 0°C, and used ethyl acetate (200mL ×3) Extract, combine the organic layers, wash with saturated sodium chloride aqueous solution (100mL×3), dry with anhydrous sodium sulfate, filter, and evaporate the filtrate under reduced pressure to obtain (4-((3-bromo-5-methyl- 1H-pyrazol-1-yl)methyl)-2-nitrobenzyl alcohol, used directly in the next reaction without purification, ESI-MS (m/z): 326.0 [M+H] + .
步骤i)：4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-2-硝基苯甲醇甲磺酸酯的制备Step i): Preparation of 4-((3-bromo-5-methyl-1H-pyrazol-1-yl)methyl)-2-nitrobenzyl alcohol methanesulfonate
将甲磺酰氯(2.85mL，36.794mmol)滴加至(4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-2-硝基苯甲醇(7.5g，22.996mmol)和三乙胺(6.4mL，45.992mmol)的二氯甲烷(70mL)溶液中，35℃反应4h，反应结束后，反应液用冰水(200mL)稀释，用二氯甲烷(150mL×3)萃取，合并有机层，饱和氯化钠水溶液(100mL×3)洗涤，无水硫酸钠干燥，过滤，滤液减压蒸干，得4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-2-硝基苯甲醇甲磺酸酯，无需纯化直接用于下一步反应。Methanesulfonyl chloride (2.85mL, 36.794mmol) was added dropwise to (4-((3-bromo-5-methyl-1H-pyrazol-1-yl)methyl)-2-nitrobenzyl alcohol (7.5g , 22.996mmol) and triethylamine (6.4mL, 45.992mmol) in dichloromethane (70mL) solution at 35℃ for 4h. After the reaction, the reaction solution was diluted with ice water (200mL) and dichloromethane (150mL ×3) Extract, combine the organic layers, wash with saturated sodium chloride aqueous solution (100mL×3), dry with anhydrous sodium sulfate, filter, and evaporate the filtrate under reduced pressure to obtain 4-((3-bromo-5-methyl-1H -Pyrazol-1-yl)methyl)-2-nitrobenzyl alcohol methanesulfonate, used directly in the next reaction without purification.
步骤j)：4-(叔丁基)-2-乙基-1-(4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-2-硝基苄基)-1H-咪唑-2,4-二羧酸的制备Step j): 4-(tert-butyl)-2-ethyl-1-(4-((3-bromo-5-methyl-1H-pyrazol-1-yl)methyl)-2-nitro Preparation of benzyl)-1H-imidazole-2,4-dicarboxylic acid
将4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-2-硝基苯甲醇甲磺酸酯(7.0g，17.316mmol)、1H-咪唑-2-甲酸乙酯-4-甲酸叔丁酯(3.96g，16.491mmol)、碳酸钾(5.98g，43.290mmol)和乙腈(70mL)加入反应瓶中，50℃搅拌反应5h，反应结束后减压蒸除溶剂，残余物加水(150mL)稀释，用乙酸乙酯(150mL×3)萃取，合并有机层，饱和氯化钠水溶液(100mL×3)洗涤，无水硫酸钠干燥，过滤，滤液减压蒸干，所得粗品经硅胶柱层析纯化(洗脱剂：石油醚/乙酸乙酯＝3/1)，得4-(叔丁基)2-乙基1-(4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-2-硝基苄基)-1H-咪唑-2,4-二羧酸，收率64.2％； 1H NMR(400MHz,CDCl 3)δ7.95(d,J＝1.6Hz,1H),7.72-7.61(m,1H),7.31-7.24(m,1H),6.69(d,J＝8.0Hz,1H),6.13(d,J＝0.8Hz,1H),6.01(s,2H),5.30(s,2H),4.37-4.29(m,2H),2.24(d,J＝0.8Hz,3H),1.61(s,9H),1.36(t,J＝7.2Hz,3H)，ESI-MS(m/z)：580.4[M+Na] +。 The 4-((3-bromo-5-methyl-1H-pyrazol-1-yl)methyl)-2-nitrobenzyl alcohol methanesulfonate (7.0g, 17.316mmol), 1H-imidazole-2 -Ethyl formate-4-carboxylic acid tert-butyl ester (3.96g, 16.491mmol), potassium carbonate (5.98g, 43.290mmol) and acetonitrile (70mL) were added to the reaction flask, stirred at 50°C for 5h, and evaporated under reduced pressure after the reaction was completed The solvent was removed, the residue was diluted with water (150 mL), extracted with ethyl acetate (150 mL×3), the organic layers were combined, washed with saturated aqueous sodium chloride solution (100 mL×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated under reduced pressure After drying, the obtained crude product was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate = 3/1) to obtain 4-(tert-butyl) 2-ethyl 1-(4-((3-bromo- 5-Methyl-1H-pyrazol-1-yl)methyl)-2-nitrobenzyl)-1H-imidazole-2,4-dicarboxylic acid, yield 64.2%; 1 H NMR (400MHz, CDCl 3 )δ7.95(d,J=1.6Hz,1H), 7.72-7.61(m,1H), 7.31-7.24(m,1H), 6.69(d,J=8.0Hz,1H), 6.13(d, J = 0.8Hz, 1H), 6.01 (s, 2H), 5.30 (s, 2H), 4.37-4.29 (m, 2H), 2.24 (d, J = 0.8 Hz, 3H), 1.61 (s, 9H), 1.36 (t, J=7.2 Hz, 3H), ESI-MS (m/z): 580.4 [M+Na] + .
步骤k)：4-(叔丁基)2-乙基1-(2-氨基-4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)苄基)-1H-咪唑-2,4-二羧酸的制备Step k): 4-(tert-butyl)2-ethyl 1-(2-amino-4-((3-bromo-5-methyl-1H-pyrazol-1-yl)methyl)benzyl) Preparation of -1H-imidazole-2,4-dicarboxylic acid
在20℃将锌粉(7.2g，109.411mmol)加入4-(叔丁基)2-乙基1-(4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-2-硝基苄基)-1H-咪唑-2,4-二羧酸(6.0g，10.941mmol)和氯化铵(5.85g，109.411mmol)的乙醇(60mL)混合液中，20℃继续搅拌反应5h，反应结束后，过滤，滤液减压浓缩，得4-(叔丁基)2-乙基1-(2-氨基-4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)苄基)-1H-咪唑-2,4-二羧酸，收率52.8％，无需纯化直接用于下一步反应， 1H NMR(400MHz,CDCl 3)δ7.6(s,1H),6.61(d,J＝7.6Hz,1H),6.43(d,J＝1.2Hz,1H),6.30(d,J＝7.6Hz,1H),6.19(s,1H),5.45-5.24(m,4H),5.10(s,2H),4.29(q,J＝7.2Hz,2H),2.19(s,3H),1.48(s,9H),1.27(t,J＝7.2Hz,3H)；ESI-MS(m/z)：520.2[M+H] +。 Add zinc powder (7.2g, 109.411mmol) to 4-(tert-butyl)2-ethyl 1-(4-((3-bromo-5-methyl-1H-pyrazol-1-yl) at 20°C (Methyl)-2-nitrobenzyl)-1H-imidazole-2,4-dicarboxylic acid (6.0g, 10.941mmol) and ammonium chloride (5.85g, 109.411mmol) in a mixture of ethanol (60mL), The reaction was stirred at 20°C for 5 hours. After the reaction, it was filtered and the filtrate was concentrated under reduced pressure to obtain 4-(tert-butyl) 2-ethyl 1-(2-amino-4-((3-bromo-5-methyl- 1H-Pyrazol-1-yl)methyl)benzyl)-1H-imidazole-2,4-dicarboxylic acid, the yield is 52.8%. It is directly used in the next reaction without purification. 1 H NMR (400MHz, CDCl 3 )δ7.6(s,1H), 6.61(d,J=7.6Hz,1H), 6.43(d,J=1.2Hz,1H), 6.30(d,J=7.6Hz,1H), 6.19(s, 1H), 5.45-5.24 (m, 4H), 5.10 (s, 2H), 4.29 (q, J = 7.2Hz, 2H), 2.19 (s, 3H), 1.48 (s, 9H), 1.27 (t, J = 7.2 Hz, 3H); ESI-MS (m/z): 520.2 [M+H] + .
步骤l)：8-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-11-氧-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000011
-2-甲酸叔丁酯的制备 Step 1): 8-((3-Bromo-5-methyl-1H-pyrazol-1-yl)methyl)-11-oxy-10,11-dihydro-5H-benzo[e]imidazole[ 1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000011
Preparation of tert-butyl -2-carboxylate
将4-(叔丁基)2-乙基1-(2-氨基-4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)苄基)-1H-咪唑-2,4-二羧酸(2.0g，3.858mmol)、叔丁醇钾(541mg，4.821mmol)和DMF(35mL)加入反应 瓶中，25℃搅拌反应2h，反应结束后，反应液在20℃用氯化铵水溶液(10mL)淬灭，用二氯甲烷(80mL×2)萃取，合并有机层，饱和氯化钠水溶液(50mL×2)洗涤，无水硫酸钠干燥，过滤，滤液减压蒸干，得白色固体，粗品收率93.2％，无需纯化直接用于下一步反应，ESI-MS(m/z)：474.0[M+H] +。 Add 4-(tert-butyl) 2-ethyl 1-(2-amino-4-((3-bromo-5-methyl-1H-pyrazol-1-yl)methyl)benzyl)-1H- Imidazole-2,4-dicarboxylic acid (2.0g, 3.858mmol), potassium tert-butoxide (541mg, 4.821mmol) and DMF (35mL) were added to the reaction flask, and the reaction was stirred at 25°C for 2h. After the reaction, the reaction solution was Quench with aqueous ammonium chloride solution (10 mL) at 20°C, extract with dichloromethane (80 mL×2), combine the organic layers, wash with saturated aqueous sodium chloride solution (50 mL×2), dry with anhydrous sodium sulfate, filter, and reduce the filtrate. It was evaporated to dryness to obtain a white solid with a crude yield of 93.2%. It was directly used in the next reaction without purification, ESI-MS (m/z): 474.0 [M+H] + .
步骤m)：8-((5-甲基-1H-吡唑-1-基)甲基)-11-氧-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000012
-2-甲酸叔丁酯的制备 Step m): 8-((5-methyl-1H-pyrazol-1-yl)methyl)-11-oxy-10,11-dihydro-5H-benzo[e]imidazole[1,2- a][1,4]diazepine
Figure PCTCN2020137640-appb-000012
Preparation of tert-butyl -2-carboxylate
将8-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-11-氧-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000013
-2-甲酸叔丁酯(1.6g，3.387mmol)、Pd/C(500mg)和HCOONH 4(2.14g，33.874mmol)和二氧六环(3mL)加入反应瓶中，45℃搅拌反应0.5h，反应结束后，减压蒸除溶剂，残余物加水(100mL)稀释，用二氯甲烷(80mL×2)萃取，合并有机层，饱和氯化钠水溶液(50mL×2)洗涤，无水硫酸钠干燥，过滤，滤液减压蒸干，得白色固体，粗品收率82.5％，无需纯化直接用于下一步反应， 1H NMR(400MHz,CDCl 3)δ10.93-10.63(m,1H),8.11-7.87(m,1H),7.44-7.27(m,2H),6.97(s,1H),6.90(d,J＝7.6Hz,1H),6.07(s,1H),5.28(d,J＝11.6Hz,4H),2.18(s,3H),1.48(s,9H)；ESI-MS(m/z)：394.1[M+H] +。 The 8-((3-bromo-5-methyl-1H-pyrazol-1-yl)methyl)-11-oxy-10,11-dihydro-5H-benzo[e]imidazole[1,2 -a][1,4]diazepine
Figure PCTCN2020137640-appb-000013
Tert-Butyl-2-carboxylate (1.6g, 3.387mmol), Pd/C (500mg) , HCOONH 4 (2.14g, 33.874mmol) and dioxane (3mL) were added to the reaction flask, stirred at 45°C for 0.5h After the reaction, the solvent was evaporated under reduced pressure, the residue was diluted with water (100mL), extracted with dichloromethane (80mL×2), the organic layers were combined, washed with saturated sodium chloride aqueous solution (50mL×2), anhydrous sodium sulfate Dry, filter, and evaporate the filtrate under reduced pressure to obtain a white solid. The crude product has a yield of 82.5%. It is directly used in the next reaction without purification. 1 H NMR (400MHz, CDCl 3 ) δ 10.93-10.63 (m, 1H), 8.11 -7.87(m,1H),7.44-7.27(m,2H),6.97(s,1H),6.90(d,J=7.6Hz,1H),6.07(s,1H), 5.28(d,J=11.6 Hz, 4H), 2.18 (s, 3H), 1.48 (s, 9H); ESI-MS (m/z): 394.1 [M+H] + .
步骤n)：8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000014
-2-甲酸叔丁酯的制备 Step n): 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1 ,4] Diaza
Figure PCTCN2020137640-appb-000014
Preparation of tert-butyl -2-carboxylate
在25℃将BH 3.THF(7.63mL，7.625mmol)滴加至8-((5-甲基-1H-吡唑-1-基)甲基)-11-氧-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000015
-2-甲酸叔丁酯(1.0g，2.542mmol)的四氢呋喃(10mL)溶液中，50℃反应3h，加甲醇(200mL)淬灭，减压浓缩，得白色固体，无需纯化直接用于下一步；ESI-MS(m/z)：380.2[M+H] +。 BH 3 .THF (7.63mL, 7.625mmol) was added dropwise to 8-((5-methyl-1H-pyrazol-1-yl)methyl)-11-oxy-10,11-dihydro at 25°C -5H-Benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000015
In a solution of tert-butyl-2-carboxylate (1.0g, 2.542mmol) in tetrahydrofuran (10mL), react at 50°C for 3h, quench with methanol (200mL), and concentrate under reduced pressure to obtain a white solid, which is used directly in the next step without purification ; ESI-MS (m/z): 380.2 [M+H] + .
步骤o)：8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000016
-2-羧酸 Step o): 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1 ,4] Diaza
Figure PCTCN2020137640-appb-000016
-2-carboxylic acid
将8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000017
-2-甲酸叔丁酯(700mg，1.845mmol)和8N盐酸(30mL)加入反应瓶中，升温至60℃反应2h，反应结束后，减压蒸除溶剂，制备HPLC纯化，得白色固体，收率60.3％；ESI-MS(m/z)：324.3[M+H] +。 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4] Diaza
Figure PCTCN2020137640-appb-000017
Tert-Butyl-2-carboxylate (700mg, 1.845mmol) and 8N hydrochloric acid (30mL) were added to the reaction flask, heated to 60°C and reacted for 2h. After the reaction, the solvent was evaporated under reduced pressure and purified by preparative HPLC to obtain a white solid. The rate is 60.3%; ESI-MS (m/z): 324.3 [M+H] + .
步骤p)：N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000018
-2-甲酰胺的制备 Step p): N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl) -10,11-Dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000018
Preparation of -2-carboxamide
将8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000019
-2-羧酸(130mg，0.361mmol)、5-(氨甲基)-4,6-二甲基吡啶-2-二胺二盐酸盐(81mg，0.361mmol)、HBTU(192mg，0.505mmol)、三乙胺(0.18mL，1.264mmol)和DMF(5mL)加入反应瓶中，室温搅拌反应1h，反应结束后，减压浓缩，残余物经制备HPLC纯化，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000020
-2-甲酰胺，收率31.2％， 1H NMR(400MHz,CDCl 3)δ8.252(s,1H),8.15(d,J＝2.8Hz,1H),7.33(d,J＝7.6Hz,1H),6.74(s,1H),6.71-6.68(m,2H),6.65(d,J＝2.8Hz,1H),5.60(s,2H),5.53(s,2H),4.75(s,2H),5.53(s,2H),2.62(s,3H),2.48(s,3H)；ESI-MS(m/z)：457.2[M+H] +。 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4] Diaza
Figure PCTCN2020137640-appb-000019
-2-carboxylic acid (130mg, 0.361mmol), 5-(aminomethyl)-4,6-lutidine-2-diamine dihydrochloride (81mg, 0.361mmol), HBTU (192mg, 0.505mmol) ), triethylamine (0.18mL, 1.264mmol) and DMF (5mL) were added to the reaction flask, stirred at room temperature for 1h, after the reaction, concentrated under reduced pressure, the residue was purified by preparative HPLC to obtain N-((6-amino -2,4-dimethylpyridin-3-yl)methyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzene And [e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000020
-2-carboxamide, yield 31.2%, 1 H NMR (400MHz, CDCl 3 ) δ 8.252 (s, 1H), 8.15 (d, J = 2.8 Hz, 1H), 7.33 (d, J = 7.6 Hz, 1H), 6.74 (s, 1H), 6.71-6.68 (m, 2H), 6.65 (d, J = 2.8Hz, 1H), 5.60 (s, 2H), 5.53 (s, 2H), 4.75 (s, 2H) ), 5.53 (s, 2H), 2.62 (s, 3H), 2.48 (s, 3H); ESI-MS (m/z): 457.2 [M+H] + .
实施例2Example 2
8-((1H-吡唑-1-基)甲基)-N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑 [1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000021
-2-甲酰胺(化合物2)的制备 8-((1H-pyrazol-1-yl)methyl)-N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-10,11-dihydro-5H -Benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000021
Preparation of -2-carboxamide (Compound 2)
Figure PCTCN2020137640-appb-000022
Figure PCTCN2020137640-appb-000022
步骤a)：4-((1H-吡唑-1-基)甲基)-2-硝基苯甲酸甲酯的制备Step a): Preparation of methyl 4-((1H-pyrazol-1-yl)methyl)-2-nitrobenzoate
将4-溴甲基-2-硝基-苯甲酸甲酯(10.0g，36.487mmol)、1H-吡唑(2.48g，36.487mmol)、碳酸钾(10.08g，72.974mmol)和乙腈(50mL)加入反应瓶中，氮气保护下50℃搅拌反应12h，反应结束后减压蒸除溶剂，残余物加水(100mL)稀释，用乙酸乙酯(100mL×2)萃取，合并有机层，饱和氯化钠水溶液(100mL)洗涤，无水硫酸钠干燥，过滤，滤液减压蒸干，所得粗品经硅胶柱层析纯化(洗脱剂：石油醚/乙酸乙酯＝2/1)，得4-((1H-吡唑-1-基)甲基)-2-硝基苯甲酸甲酯，收率83.9％， 1H NMR(400MHz,CDCl 3)δ7.75-7.69(m,1H),7.68-7.64(m,1H),7.63-7.58(m,1H),7.50-7.47(m,1H),7.46-7.41(m,1H),6.40-6.30(m,1H),5.47-5.41(m,2H),3.95-3.86(m,3H)；ESI-MS(m/z)：262.1[M+H] +。 Combine 4-bromomethyl-2-nitro-benzoic acid methyl ester (10.0g, 36.487mmol), 1H-pyrazole (2.48g, 36.487mmol), potassium carbonate (10.08g, 72.974mmol) and acetonitrile (50mL) Add to the reaction flask, stir the reaction at 50°C under nitrogen protection for 12h. After the reaction, the solvent is evaporated under reduced pressure. The residue is diluted with water (100mL) and extracted with ethyl acetate (100mL×2). The organic layers are combined and saturated sodium chloride. Wash with aqueous solution (100 mL), dry with anhydrous sodium sulfate, filter, and evaporate the filtrate under reduced pressure. The crude product obtained is purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate = 2/1) to give 4-(( 1H-pyrazol-1-yl)methyl)-2-nitrobenzoic acid methyl ester, yield 83.9%, 1 H NMR (400MHz, CDCl 3 )δ7.75-7.69(m,1H),7.68-7.64 (m,1H),7.63-7.58(m,1H),7.50-7.47(m,1H),7.46-7.41(m,1H),6.40-6.30(m,1H),5.47-5.41(m,2H) , 3.95-3.86 (m, 3H); ESI-MS (m/z): 262.1 [M+H] + .
步骤b)：4-((1H-吡唑-1-基)甲基)-2-硝基苯甲醇的制备Step b): Preparation of 4-((1H-pyrazol-1-yl)methyl)-2-nitrobenzyl alcohol
在0℃将硼氢化钠(2.32g，61.248mmol)和氯化钙(2.55g，22.968mmol)加入4-((1H-吡唑-1-基)甲基)-2-硝基苯甲酸甲酯(4.0g，15.312mmol)的四氢呋喃(40mL)溶液中，0℃搅拌反应5h，反应液在0℃用氯化铵水溶液(100mL)淬灭，用乙酸乙酯(200mL×2)萃取，合并有机层，饱和氯化钠水溶液(100mL)洗涤，无水硫酸钠干燥，过滤，滤液减压蒸干，所得粗品用甲基叔丁基醚打浆纯化，得4-((1H-吡唑-1-基)甲基)-2-硝基苯甲醇，收率82.3％， 1H NMR(400MHz,CDCl 3)δ7.93-7.89(m,1H),7.77-7.71(m,1H),7.62-7.57(m,1H),7.49-7.43(m,2H),6.38-6.33(m,1H),5.45-5.38(m,2H),4.99-4.93(m,2H)，ESI-MS(m/z)：234.1[M+H] +。 Add sodium borohydride (2.32g, 61.248mmol) and calcium chloride (2.55g, 22.968mmol) to 4-((1H-pyrazol-1-yl)methyl)-2-nitrobenzoic acid methyl at 0°C In the tetrahydrofuran (40mL) solution of the ester (4.0g, 15.312mmol), the reaction was stirred at 0°C for 5h. The reaction solution was quenched with ammonium chloride aqueous solution (100mL) at 0°C, extracted with ethyl acetate (200mL×2), and combined The organic layer was washed with saturated aqueous sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure. The crude product obtained was purified by beating with methyl tert-butyl ether to obtain 4-((1H-pyrazole-1) -Yl)methyl)-2-nitrobenzyl alcohol, yield 82.3%, 1 H NMR (400MHz, CDCl 3 ) δ7.93-7.89 (m, 1H), 7.77-7.71 (m, 1H), 7.62 7.57(m,1H),7.49-7.43(m,2H),6.38-6.33(m,1H),5.45-5.38(m,2H),4.99-4.93(m,2H), ESI-MS(m/z ): 234.1[M+H] + .
步骤c)：4-((1H-吡唑-1-基)甲基)-2-硝基苯甲醇甲磺酸酯的制备Step c): Preparation of 4-((1H-pyrazol-1-yl)methyl)-2-nitrobenzyl alcohol methanesulfonate
将甲磺酰氯(0.786mL，10.146mmol)滴加至4-((1H-吡唑-1-基)甲基)-2-硝基苯甲醇(1.5g，6.431mmol)和三乙胺(1.79mL，12.862mmol)的二氯甲烷(15mL)溶液中，35℃反应8h，反应结束后，将反应液倾入冰水(50mL)中，用二氯甲烷(50mL×3)萃取，合并 有机层，饱和氯化钠水溶液(100mL×3)洗涤，无水硫酸钠干燥，过滤，滤液减压蒸干，得4-((1H-吡唑-1-基)甲基)-2-硝基苯甲醇甲磺酸酯，无需纯化直接用于下一步反应。Methanesulfonyl chloride (0.786mL, 10.146mmol) was added dropwise to 4-((1H-pyrazol-1-yl)methyl)-2-nitrobenzyl alcohol (1.5g, 6.431mmol) and triethylamine (1.79 mL, 12.862mmol) in dichloromethane (15mL) solution, react at 35℃ for 8h. After the reaction is over, pour the reaction solution into ice water (50mL), extract with dichloromethane (50mL×3), and combine the organic layers , Washed with saturated sodium chloride aqueous solution (100mL×3), dried with anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain 4-((1H-pyrazol-1-yl)methyl)-2-nitrobenzene Methanol mesylate is used directly in the next reaction without purification.
步骤d)：4-(叔丁基)-2-乙基-1-(4-((1H-吡唑-1-基)甲基)-2-硝基苄基)-1H-咪唑-2,4-二羧酸的制备Step d): 4-(tert-butyl)-2-ethyl-1-(4-((1H-pyrazol-1-yl)methyl)-2-nitrobenzyl)-1H-imidazole-2 Preparation of ,4-dicarboxylic acid
将4-((1H-吡唑-1-基)甲基)-2-硝基苯甲醇甲磺酸酯(1.8g，5.782mmol)、1H-咪唑-2-甲酸乙酯-4-甲酸叔丁酯(1.32g，5.493mmol)、碳酸钾(2.0g，14.455mmol)和乙腈(9mL)加入反应瓶中，50℃搅拌反应5h，反应结束后减压蒸除溶剂，残余物加水(10mL)稀释，用乙酸乙酯(20mL×3)萃取，合并有机层，饱和氯化钠水溶液(20mL)洗涤，无水硫酸钠干燥，过滤，滤液减压蒸干，所得粗品经硅胶柱层析纯化(洗脱剂：石油醚/乙酸乙酯＝1/1)，得4-(叔丁基)2-乙基1-(4-((1H-吡唑-1-基)甲基)-2-硝基苄基)-1H-咪唑-2,4-二羧酸，收率36.8％，ESI-MS(m/z)：456.2[M+H] +。 Combine 4-((1H-pyrazol-1-yl)methyl)-2-nitrobenzyl alcohol methanesulfonate (1.8g, 5.772mmol), 1H-imidazole-2-carboxylic acid ethyl ester-4-carboxylic acid tert Butyl ester (1.32g, 5.493mmol), potassium carbonate (2.0g, 14.455mmol) and acetonitrile (9mL) were added to the reaction flask, and the reaction was stirred at 50°C for 5h. After the reaction, the solvent was evaporated under reduced pressure, and the residue was added with water (10mL) Dilute, extract with ethyl acetate (20mL×3), combine the organic layers, wash with saturated aqueous sodium chloride (20mL), dry with anhydrous sodium sulfate, filter, and evaporate the filtrate under reduced pressure to dryness. The resulting crude product is purified by silica gel column chromatography ( Eluent: petroleum ether/ethyl acetate=1/1) to give 4-(tert-butyl)2-ethyl 1-(4-((1H-pyrazol-1-yl)methyl)-2- Nitrobenzyl)-1H-imidazole-2,4-dicarboxylic acid, yield 36.8%, ESI-MS (m/z): 456.2 [M+H] + .
步骤e)：4-(叔丁基)2-乙基1-(4-((1H-吡唑-1-基)甲基)-2-氨基苄基)-1H-咪唑-2,4-二羧酸的制备Step e): 4-(tert-butyl)2-ethyl 1-(4-((1H-pyrazol-1-yl)methyl)-2-aminobenzyl)-1H-imidazole-2,4- Preparation of dicarboxylic acid
在20℃将锌粉(1.72g，26.346mmol)加入4-(叔丁基)2-乙基1-(4-((1H-吡唑-1-基)甲基)-2-硝基苄基)-1H-咪唑-2,4-二羧酸(1.2g，2.635mmol)和氯化铵(1.41g，26.346mmol)的乙醇(40mL)混合液中，25℃继续搅拌反应2h，反应结束后，过滤，滤液减压浓缩，所得粗品经制备型HPLC纯化，得4-(叔丁基)2-乙基1-(4-((1H-吡唑-1-基)甲基)-2-氨基苄基)-1H-咪唑-2,4-二羧酸，收率44.6％， 1H NMR(400MHz,DMSO-d 6)δ7.74-7.72(m,1H),7.71-7.69(m,1H),7.44-7.41(m,1H),6.63-6.59(m,1H),6.51-6.48(m,1H),6.39-6.33(m,1H),6.27-6.22(m,1H),5.42-5.37(m,2H),5.33-5.27(m,2H),5.18-5.13(m,2H),4.33-4.25(m,2H),1.60-1.40(m,9H),1.30-1.24(m,3H)；ESI-MS(m/z)：426.3[M+H] +。 Add zinc powder (1.72g, 26.346mmol) to 4-(tert-butyl)2-ethyl 1-(4-((1H-pyrazol-1-yl)methyl)-2-nitrobenzyl at 20℃ Base)-1H-imidazole-2,4-dicarboxylic acid (1.2g, 2.635mmol) and ammonium chloride (1.41g, 26.346mmol) in ethanol (40mL) mixture, stirring at 25℃ for 2h, the reaction is complete After filtration, the filtrate was concentrated under reduced pressure, and the resulting crude product was purified by preparative HPLC to obtain 4-(tert-butyl)2-ethyl 1-(4-((1H-pyrazol-1-yl)methyl)-2 -Aminobenzyl)-1H-imidazole-2,4-dicarboxylic acid, yield 44.6%, 1 H NMR (400MHz, DMSO-d 6 )δ7.74-7.72(m,1H),7.71-7.69(m ,1H),7.44-7.41(m,1H),6.63-6.59(m,1H),6.51-6.48(m,1H),6.39-6.33(m,1H),6.27-6.22(m,1H),5.42 -5.37(m,2H),5.33-5.27(m,2H),5.18-5.13(m,2H),4.33-4.25(m,2H),1.60-1.40(m,9H),1.30-1.24(m, 3H); ESI-MS (m/z): 426.3 [M+H] + .
步骤f)：8-((1H-吡唑-1-基)甲基)-11-氧-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000023
-2-甲酸叔丁酯的制备 Step f): 8-((1H-pyrazol-1-yl)methyl)-11-oxy-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1, 4] Diaza
Figure PCTCN2020137640-appb-000023
Preparation of tert-butyl -2-carboxylate
将4-(叔丁基)2-乙基1-(4-((1H-吡唑-1-基)甲基)-2-氨基苄基)-1H-咪唑-2,4-二羧酸(500mg，1.175mmol)、叔丁醇钾(165mg，1.469mmol)和DMF(5mL)加入反应瓶中，25℃搅拌反应10h，反应结束后，加水(20mL)淬灭，用二氯甲烷(20mL×4)萃取，合并有机层，饱和氯化钠水溶液(10mL)洗涤，无水硫酸钠干燥，过滤，滤液减压蒸干，得白色固体，粗品收率67.2％，无需纯化直接用于下一步反应， 1H NMR(400MHz,DMSO-d 6)δ10.87-10.71(m,1H),8.56-8.54(m,1H),7.99-7.97(m,1H),7.80-7.76(m,1H),7.46-7.43(m,1H),7.36-7.32(m,1H),7.05-7.02(m,1H),6.99-6.95(m,1H),6.27-6.24(m,1H),5.32-5.29(m,2H),5.28-5.24(m,2H),1.49-1.45(m,9H)；ESI-MS(m/z)：380.3[M+H] +。 Add 4-(tert-butyl)2-ethyl 1-(4-((1H-pyrazol-1-yl)methyl)-2-aminobenzyl)-1H-imidazole-2,4-dicarboxylic acid (500mg, 1.175mmol), potassium tert-butoxide (165mg, 1.469mmol) and DMF (5mL) were added to the reaction flask, and the reaction was stirred at 25°C for 10h. After the reaction was completed, quenched with water (20mL), and dichloromethane (20mL) ×4) Extract, combine the organic layers, wash with saturated sodium chloride aqueous solution (10 mL), dry with anhydrous sodium sulfate, filter, and evaporate the filtrate under reduced pressure to obtain a white solid. The crude product has a yield of 67.2%. It is directly used in the next step without purification. Reaction, 1 H NMR (400MHz, DMSO-d 6 ) δ 10.87-10.71 (m, 1H), 8.56-8.54 (m, 1H), 7.99-7.97 (m, 1H), 7.80-7.76 (m, 1H) ,7.46-7.43(m,1H),7.36-7.32(m,1H),7.05-7.02(m,1H),6.99-6.95(m,1H),6.27-6.24(m,1H),5.32-5.29( m, 2H), 5.28-5.24 (m, 2H), 1.49-1.45 (m, 9H); ESI-MS (m/z): 380.3 [M+H] + .
步骤g)：8-((1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000024
-2-甲酸叔丁酯的制备 Step g): 8-((1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine miscellaneous
Figure PCTCN2020137640-appb-000024
Preparation of tert-butyl -2-carboxylate
在25℃将BH 3.THF(2.64mL，2.635mmol)滴加至8-((1H-吡唑-1-基)甲基)-11-氧-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000025
-2-甲酸叔丁酯(200mg，0.527mmol)的四氢呋喃(2mL)溶液中，50℃反应2h，加甲醇(5mL)淬灭，减压浓缩，得黄色固体，无需纯化直接用于下一步；ESI-MS(m/z)：366.2[M+H] +。 BH 3 .THF (2.64mL, 2.635mmol) was added dropwise to 8-((1H-pyrazol-1-yl)methyl)-11-oxy-10,11-dihydro-5H-benzo at 25°C [e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000025
In a solution of tert-butyl-2-carboxylate (200mg, 0.527mmol) in tetrahydrofuran (2mL), react at 50°C for 2h, quench with methanol (5mL), and concentrate under reduced pressure to obtain a yellow solid, which is used directly in the next step without purification; ESI-MS (m/z): 366.2 [M+H] + .
步骤h)：8-((1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000026
-2-羧酸的制备 Step h): 8-((1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine miscellaneous
Figure PCTCN2020137640-appb-000026
Preparation of -2-carboxylic acid
将8-((1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000027
-2-甲酸叔丁酯(150mg，0.410mmol)和8N盐酸(2.05mL)加入反应瓶中，升温至60℃反应2h，反应结束后，减压蒸除溶剂，制备HPLC纯化，得8-((1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并 [e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000028
-2-羧酸，收率70.4％；ESI-MS(m/z)：310.2[M+H] +。 8-((1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000027
Tert-Butyl-2-carboxylate (150mg, 0.410mmol) and 8N hydrochloric acid (2.05mL) were added to the reaction flask, heated to 60°C and reacted for 2h. After the reaction, the solvent was evaporated under reduced pressure and purified by preparative HPLC to obtain 8-( (1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000028
-2-carboxylic acid, the yield is 70.4%; ESI-MS (m/z): 310.2 [M+H] + .
步骤i)：8-((1H-吡唑-1-基)甲基)-N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000029
-2-甲酰胺的制备 Step i): 8-((1H-pyrazol-1-yl)methyl)-N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-10,11- Dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000029
Preparation of -2-carboxamide
将8-((1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000030
-2-羧酸(100mg，0.289mmol)、5-(氨甲基)-4,6-二甲基吡啶-2-二胺二盐酸盐(71mg，0.318mmol)、HBTU(153mg，0.404mmol)、三乙胺(0.12mL，0.867mmol)和DMF(1mL)加入反应瓶中，室温搅拌反应1h，反应结束后，减压浓缩，残余物经制备HPLC纯化，得8-((1H-吡唑-1-基)甲基)-N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000031
-2-甲酰胺，收率41.9％， 1H NMR(400MHz,DMSO-d 6)δ13.56-13.45(m,1H),8.30-8.22(m,1H),7.76-7.74(m,1H),7.73-7.71(m,1H),7.68-7.60(m,2H),7.46-7.40(m,1H),7.00-6.96(m,1H),6.61-6.59(m,1H),6.43-6.39(m,2H),6.25-6.22(m,1H),5.33-5.30(m,2H),5.15-5.13(m,2H),4.43-4.41(m,2H),4.30-4.28(m,2H),2.51-2.51(m,3H),2.38-2.35(m,3H)；ESI-MS(m/z)：443.2[M+H] +。 8-((1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000030
-2-carboxylic acid (100mg, 0.289mmol), 5-(aminomethyl)-4,6-lutidine-2-diamine dihydrochloride (71mg, 0.318mmol), HBTU (153mg, 0.404mmol) ), triethylamine (0.12mL, 0.867mmol) and DMF (1mL) were added to the reaction flask, stirred at room temperature for 1h, after the reaction, concentrated under reduced pressure, the residue was purified by preparative HPLC to obtain 8-((1H-pyridine (Azol-1-yl)methyl)-N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-10,11-dihydro-5H-benzo(e)imidazole [1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000031
-2-carboxamide, yield 41.9%, 1 H NMR (400MHz, DMSO-d 6 ) δ 13.56-13.45 (m, 1H), 8.30-8.22 (m, 1H), 7.76-7.74 (m, 1H) ,7.73-7.71(m,1H),7.68-7.60(m,2H),7.46-7.40(m,1H),7.00-6.96(m,1H),6.61-6.59(m,1H),6.43-6.39( m,2H),6.25-6.22(m,1H),5.33-5.30(m,2H),5.15-5.13(m,2H),4.43-4.41(m,2H),4.30-4.28(m,2H), 2.51-2.51 (m, 3H), 2.38-2.35 (m, 3H); ESI-MS (m/z): 443.2 [M+H] + .
实施例3Example 3
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((4-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000032
-2-甲酰胺(化合物3)的制备 N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((4-methyl-1H-pyrazol-1-yl)methyl)-10,11 -Dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000032
Preparation of -2-carboxamide (Compound 3)
Figure PCTCN2020137640-appb-000033
Figure PCTCN2020137640-appb-000033
操作过程同实施例2，只是将步骤a中起始原料1H-吡唑用4-甲基-1H-吡唑替代，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((4-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000034
-2-甲酰胺， 1H NMR(400MHz,DMSO-d 6)δ8.18(brs,1H),7.70(s,1H),7.45-7.44(m,3H),7.26(s,1H),6.96(d,J＝7.6Hz,1H),6.60(s,1H),6.41-6.38(m,2H),5.30(s,2H),5.04(s,2H),4.40(s,2H),4.28(d,J＝6.0Hz,2H),2.51(s,3H),2.37(s,3H),1.97(s,3H)；ESI-MS(m/z)：457.1[M+H] +。 The operation process is the same as in Example 2, except that the starting material 1H-pyrazole in step a is replaced by 4-methyl-1H-pyrazole to obtain N-((6-amino-2,4-lutidine-3 -Yl)methyl)-8-((4-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a ][1,4]diazepine
Figure PCTCN2020137640-appb-000034
-2-Carboxamide, 1 H NMR(400MHz,DMSO-d 6 )δ8.18(brs,1H),7.70(s,1H),7.45-7.44(m,3H),7.26(s,1H),6.96 (d,J=7.6Hz,1H),6.60(s,1H),6.41-6.38(m,2H),5.30(s,2H),5.04(s,2H),4.40(s,2H),4.28( d, J=6.0 Hz, 2H), 2.51 (s, 3H), 2.37 (s, 3H), 1.97 (s, 3H); ESI-MS (m/z): 457.1 [M+H] + .
实施例4Example 4
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((4-氟-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000035
-2-甲酰胺(化合物4)的制备 N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((4-fluoro-1H-pyrazol-1-yl)methyl)-10,11- Dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000035
Preparation of -2-carboxamide (Compound 4)
Figure PCTCN2020137640-appb-000036
Figure PCTCN2020137640-appb-000036
操作过程同实施例2，只是将步骤a中起始原料1H-吡唑用4-氟-1H-吡唑替代，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((4-氟-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000037
-2-甲酰胺， 1H NMR(400MHz,DMSO-d 6)δ8.20(brs,1H),8.01(s,1H),7.95(s,1H),7.50(d,J＝3.6Hz,1H),7.39(d,J＝7.6Hz,1H),7.07-7.01(m,2H),6.50(s,1H),6.32-6.29(m,2H),5.32(s,2H),5.14(s,2H),4.51(s,2H),4.28(d,J＝6.0Hz,2H),2.51(s,3H), 2.37(s,3H)；ESI-MS(m/z)：461.3[M+H] +。 The operation process is the same as in Example 2, except that the starting material 1H-pyrazole in step a is replaced with 4-fluoro-1H-pyrazole to obtain N-((6-amino-2,4-dimethylpyridine-3- Yl)methyl)-8-((4-fluoro-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][ 1,4] Diaza
Figure PCTCN2020137640-appb-000037
-2-Carboxamide, 1 H NMR(400MHz,DMSO-d 6 )δ8.20(brs,1H),8.01(s,1H),7.95(s,1H),7.50(d,J=3.6Hz,1H ), 7.39 (d, J = 7.6 Hz, 1H), 7.07-7.01 (m, 2H), 6.50 (s, 1H), 6.32-6.29 (m, 2H), 5.32 (s, 2H), 5.14 (s, 2H),4.51(s,2H),4.28(d,J=6.0Hz,2H),2.51(s,3H), 2.37(s,3H); ESI-MS(m/z): 461.3[M+H ] + .
实施例5Example 5
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((3,5-二甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000038
-2-甲酰胺(化合物5)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-8-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)- 10,11-Dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000038
Preparation of -2-carboxamide (Compound 5)
Figure PCTCN2020137640-appb-000039
Figure PCTCN2020137640-appb-000039
操作过程同实施例2，只是将步骤a中起始原料1H-吡唑用3,5-二甲基-1H-吡唑替代，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((3,5-二甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000040
-2-甲酰胺， 1H NMR(400MHz,MeOD)δ7.79(s,1H),7.12(d,J＝7.6Hz,1H),6.68(s,1H),6.50-6.45(m,2H),5.96(s,1H),5.35(s,2H),5.14(s,2H),4.51(s,2H),4.48(s,2H),2.58(s,3H),2.46(s,3H),2.20(s,3H),2.18(s,3H)；ESI-MS(m/z)：471.3[M+H] +。 The operation process is the same as that in Example 2, except that the starting material 1H-pyrazole in step a is replaced with 3,5-dimethyl-1H-pyrazole to obtain N-((6-amino-2,4-dimethyl (Pyridin-3-yl)methyl)-8-((3,5-Dimethyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo(e)imidazole [1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000040
-2-Carboxamide, 1 H NMR (400MHz, MeOD) δ 7.79 (s, 1H), 7.12 (d, J = 7.6 Hz, 1H), 6.68 (s, 1H), 6.50-6.45 (m, 2H) , 5.96(s, 1H), 5.35(s, 2H), 5.14(s, 2H), 4.51(s, 2H), 4.48(s, 2H), 2.58(s, 3H), 2.46(s, 3H), 2.20 (s, 3H), 2.18 (s, 3H); ESI-MS (m/z): 471.3 [M+H] + .
实施例6Example 6
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((2-氧代吡啶-1(2H)-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000041
-2-甲酰胺(化合物6)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-8-((2-oxopyridin-1(2H)-yl)methyl)-10,11- Dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000041
Preparation of -2-carboxamide (Compound 6)
Figure PCTCN2020137640-appb-000042
Figure PCTCN2020137640-appb-000042
操作过程同实施例2，只是将步骤a中起始原料1H-吡唑用吡啶-2(1H)-酮替代，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((2-氧代吡啶-1(2H)-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000043
-2-甲酰胺， 1H NMR(400MHz,DMSO-d 6)δ8.25(t,J＝5.2Hz,1H),7.73(s,1H),7.64(dd,J 1＝1.8,J 2＝6.8Hz,1H),7.59(brs,2H),7.40(m,1H),6.98(d,J＝7.6Hz,1H),6.60(s,1H),6.49-6.45(m,2H),6.38(d,J＝8.8Hz,1H),6.20(dt,J 1＝1.2,J 2＝6.7Hz,1H),5.31(s,2H),4.91(s,2H),4.41(s,2H),4.29(d,J＝5.2Hz,2H),2.49-2.49(m,3H),2.36(s,3H)；ESI-MS(m/z)：470.2[M+H] +。 The operation process is the same as in Example 2, except that the starting material 1H-pyrazole in step a is replaced with pyridine-2(1H)-one to obtain N-((6-amino-2,4-dimethylpyridine-3- Yl)methyl)-8-((2-oxopyridine-1(2H)-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][ 1,4] Diaza
Figure PCTCN2020137640-appb-000043
-2-Carboxamide, 1 H NMR (400MHz, DMSO-d 6 ) δ 8.25 (t, J = 5.2 Hz, 1H), 7.73 (s, 1H), 7.64 (dd, J 1 ＝1.8, J 2 ＝ 6.8Hz, 1H), 7.59 (brs, 2H), 7.40 (m, 1H), 6.98 (d, J = 7.6 Hz, 1H), 6.60 (s, 1H), 6.49-6.45 (m, 2H), 6.38 ( d, J = 8.8Hz, 1H), 6.20 (dt, J 1 = 1.2, J 2 = 6.7Hz, 1H), 5.31(s, 2H), 4.91(s, 2H), 4.41(s, 2H), 4.29 (d, J=5.2 Hz, 2H), 2.49-2.49 (m, 3H), 2.36 (s, 3H); ESI-MS (m/z): 470.2 [M+H] + .
实施例7Example 7
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((2-氧代噁唑烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000044
-2-甲酰胺(化合物7)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-8-((2-oxazolidin-3-yl)methyl)-10,11-bis Hydrogen-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000044
Preparation of -2-carboxamide (Compound 7)
Figure PCTCN2020137640-appb-000045
Figure PCTCN2020137640-appb-000045
操作过程同实施例2，只是将步骤a中起始原料1H-吡唑用噁唑烷-2-酮替代，将碳酸钾用NaH替代，将乙腈用四氢呋喃替代，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((2-氧代噁唑烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000046
-2-甲酰胺； 1H NMR(400MHz,MeOD)δ7.68(s,1H),7.10(d,J＝8.0Hz,1H),6.58(s,1H),6.48-6.40(m,2H),5.84(s,1H),5.35 (s,2H),5.10(s,2H),4.53(dd,J 1＝7.6,J 2＝6.4Hz,2H),4.49(s,2H),4.31(s,2H),4.13(dd,J 1＝7.6,J 2＝6.4Hz,2H),2.58(s,3H),2.41(s,3H)；ESI-MS(m/z)：462.3[M+H] +。 The operation process is the same as in Example 2, except that the starting material 1H-pyrazole in step a is replaced by oxazolidin-2-one, potassium carbonate is replaced by NaH, and acetonitrile is replaced by tetrahydrofuran to obtain N-((6-amino -2,4-dimethylpyridin-3-yl)methyl)-8-((2-oxazolidin-3-yl)methyl)-10,11-dihydro-5H-benzo[ e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000046
-2-Carboxamide; 1 H NMR (400MHz, MeOD) δ 7.68 (s, 1H), 7.10 (d, J = 8.0 Hz, 1H), 6.58 (s, 1H), 6.48-6.40 (m, 2H) ,5.84(s,1H),5.35 (s,2H),5.10(s,2H),4.53(dd,J 1 =7.6,J 2 =6.4Hz,2H),4.49(s,2H),4.31(s ,2H),4.13(dd,J 1 =7.6,J 2 =6.4Hz,2H),2.58(s,3H),2.41(s,3H); ESI-MS(m/z): 462.3[M+H ] + .
实施例8Example 8
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000047
-2-甲酰胺(化合物8)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-8-((2-oxo-3-azabicyclo[3.1.0]hexane-3-yl )Methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000047
Preparation of -2-carboxamide (Compound 8)
Figure PCTCN2020137640-appb-000048
Figure PCTCN2020137640-appb-000048
操作过程同实施例2，只是将步骤a中起始原料1H-吡唑用3-氮杂双环[3.1.0]己-2-酮替代，将碳酸钾用NaH替代，将乙腈用四氢呋喃替代，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000049
-2-甲酰胺； 1H NMR(400MHz,MeOD)δ7.91(s,1H),7.18(d,J＝7.6Hz,1H),6.68(s,1H),6.60(d,J＝7.6Hz,1H),6.45(s,1H),5.37(s,2H),5.09(s,2H),4.50(s,2H),4.39-4.36(m,2H),3.62-3.48(m,1H),3.17-3.12(m,1H),2.58(s,3H),2.46(s,3H),1.99-1.96(m,1H),1.83-1.78(m,1H),1.10-1.03(m,1H),0.58-0.53(m,1H)；ESI-MS(m/z)：472.2[M+H] +。 The operation process is the same as in Example 2, except that the starting material 1H-pyrazole in step a is replaced by 3-azabicyclo[3.1.0]hexan-2-one, potassium carbonate is replaced by NaH, and acetonitrile is replaced by tetrahydrofuran. Get N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((2-oxo-3-azabicyclo[3.1.0]hexane-3- (Yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000049
-2-Carboxamide; 1 H NMR(400MHz, MeOD)δ7.91(s,1H), 7.18(d,J=7.6Hz,1H), 6.68(s,1H), 6.60(d,J=7.6Hz ,1H),6.45(s,1H),5.37(s,2H),5.09(s,2H),4.50(s,2H),4.39-4.36(m,2H),3.62-3.48(m,1H), 3.17-3.12(m,1H),2.58(s,3H),2.46(s,3H),1.99-1.96(m,1H),1.83-1.78(m,1H),1.10-1.03(m,1H), 0.58-0.53 (m, 1H); ESI-MS (m/z): 472.2 [M+H] + .
实施例9Example 9
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-苯胺基-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000050
-2-甲酰胺(化合物9)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-8-anilino-10,11-dihydro-5H-benzo[e]imidazole[1,2- a][1,4]diazepine
Figure PCTCN2020137640-appb-000050
Preparation of -2-carboxamide (Compound 9)
Figure PCTCN2020137640-appb-000051
Figure PCTCN2020137640-appb-000051
步骤a)：2-硝基-4-(苯胺基)苯甲酸甲酯的制备Step a): Preparation of methyl 2-nitro-4-(anilino)benzoate
将4-溴-2-硝基苯甲酸甲酯(5.0g，19.228mmol)、苯胺(1.97g，21.151mmol)、碳酸铯(9.40g，28.842mmol)、二环己基[3,6-二甲氧基-2',4',6'-三异丙基[1,1'-联苯]-2-基]膦 (BrettPhos，516mg，0.961mmol)和Pd 2(dba) 3(880mg，0.961mmol)加入反应瓶中，氩气置换出反应瓶中空气后加无水四氢呋喃(100mL)，加毕，氩气保护下升温至回流反应2h，反应结束后，减压浓缩，残余物加水稀释，乙酸乙酯萃取，合并有机层，饱和氯化钠溶液洗涤两次，无水硫酸钠干燥，过滤，滤液减压浓缩，残余物经硅胶柱层析纯化(洗脱剂：石油醚/乙酸乙酯＝10/1)，得2-硝基-4-(苯胺基)苯甲酸甲酯，收率75.6％，ESI-MS(m/z)：273.1[M+H] +。步骤b-i)：N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-苯胺基-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000052
-2-甲酰胺的制备 Methyl 4-bromo-2-nitrobenzoate (5.0g, 19.228mmol), aniline (1.97g, 21.151mmol), cesium carbonate (9.40g, 28.842mmol), dicyclohexyl[3,6-dimethyl Oxy-2',4',6'-triisopropyl[1,1'-biphenyl]-2-yl]phosphine (BrettPhos, 516mg, 0.961mmol) and Pd 2 (dba) 3 (880mg, 0.961 mmol) was added to the reaction flask, argon replaced the air in the reaction flask and then anhydrous tetrahydrofuran (100mL) was added. After the addition, the temperature was raised to reflux for 2h under the protection of argon. After the reaction was completed, it was concentrated under reduced pressure, and the residue was diluted with water. Extract with ethyl acetate, combine the organic layers, wash twice with saturated sodium chloride solution, dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify the residue by silica gel column chromatography (eluent: petroleum ether/ethyl acetate =10/1) to obtain methyl 2-nitro-4-(anilino)benzoate with a yield of 75.6%, ESI-MS (m/z): 273.1 [M+H] + . Step bi): N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-anilino-10,11-dihydro-5H-benzo[e]imidazole[ 1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000052
Preparation of -2-carboxamide
操作过程同实施例2中的步骤b-i，只是将实施例2中的4-((1H-吡唑-1-基)甲基)-2-硝基苯甲酸甲酯用相应的2-硝基-4-(苯胺基)苯甲酸甲酯替代，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-苯胺基-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000053
-2-甲酰胺， 1H NMR(400MHz,MeOD)δ8.02(s,1H),7.40(d,J＝8.0Hz,2H),7.12(d,J＝7.6Hz,1H),7.08-7.04(m,3H),6.68(s,1H),6.50-6.45(m,2H),5.30(s,2H),4.89(s,2H),4.47(s,2H),2.51-49(m,3H),2.41(m,3H)；ESI-MS(m/z)：454.3[M+H] +。 The operation process is the same as the step bi in Example 2, except that the methyl 4-((1H-pyrazol-1-yl)methyl)-2-nitrobenzoate in Example 2 is used with the corresponding 2-nitro -4-(anilino)methyl benzoate is substituted to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-anilino-10,11-dihydro -5H-Benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000053
-2-Carboxamide, 1 H NMR (400MHz, MeOD) δ 8.02 (s, 1H), 7.40 (d, J = 8.0 Hz, 2H), 7.12 (d, J = 7.6 Hz, 1H), 7.08-7.04 (m, 3H), 6.68 (s, 1H), 6.50-6.45 (m, 2H), 5.30 (s, 2H), 4.89 (s, 2H), 4.47 (s, 2H), 2.51-49 (m, 3H) ), 2.41 (m, 3H); ESI-MS (m/z): 454.3 [M+H] + .
实施例10Example 10
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-苯氧基-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000054
-2-甲酰胺(化合物10)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-8-phenoxy-10,11-dihydro-5H-benzo[e]imidazole[1,2 -a][1,4]diazepine
Figure PCTCN2020137640-appb-000054
Preparation of -2-carboxamide (Compound 10)
Figure PCTCN2020137640-appb-000055
Figure PCTCN2020137640-appb-000055
操作过程同实施例9，只是分别将步骤a中的苯胺用苯酚替代，碳酸铯用磷酸钾替代，二环己基[3,6-二甲氧基-2',4',6'-三异丙基[1,1'-联苯]-2-基]膦用(2-联苯基)二叔丁基膦替代，Pd 2(dba) 3用Pd(OAc) 2替代，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-苯氧基-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000056
-2-甲酰胺； 1H NMR(400MHz,MeOD)δ8.19(s,1H),7.42(d,J＝8.0Hz,2H),7.16(d,J＝7.6Hz,1H),7.08-7.04(m,3H),6.65(s,1H),6.50-6.45(m,2H),5.27(s,2H),4.81(s,2H),4.27(s,2H),2.56(s,3H),2.40(s,3H)；ESI-MS(m/z)：455.3[M+H] +。 The operation process is the same as in Example 9, except that the aniline in step a is replaced by phenol, the cesium carbonate is replaced by potassium phosphate, and the dicyclohexyl[3,6-dimethoxy-2',4',6'-triisopropyl Propyl[1,1'-biphenyl]-2-yl]phosphine is replaced by (2-biphenyl)di-tert-butylphosphine, and Pd 2 (dba) 3 is replaced by Pd(OAc) 2 to give N-( (6-Amino-2,4-dimethylpyridin-3-yl)methyl)-8-phenoxy-10,11-dihydro-5H-benzo(e)imidazole[1,2-a] [1,4] Diaza
Figure PCTCN2020137640-appb-000056
-2-Carboxamide; 1 H NMR (400MHz, MeOD) δ 8.19 (s, 1H), 7.42 (d, J = 8.0 Hz, 2H), 7.16 (d, J = 7.6 Hz, 1H), 7.08-7.04 (m, 3H), 6.65 (s, 1H), 6.50-6.45 (m, 2H), 5.27 (s, 2H), 4.81 (s, 2H), 4.27 (s, 2H), 2.56 (s, 3H), 2.40 (s, 3H); ESI-MS (m/z): 455.3 [M+H] + .
实施例11Example 11
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-(苄胺基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000057
-2-甲酰胺(化合物11)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-8-(benzylamino)-10,11-dihydro-5H-benzo[e]imidazole[1 ,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000057
Preparation of -2-carboxamide (Compound 11)
Figure PCTCN2020137640-appb-000058
Figure PCTCN2020137640-appb-000058
操作过程同实施例9，只是分别将步骤a中的苯胺用苄胺替代，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-(苄胺基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000059
-2-甲酰胺， 1HNMR(400MHz,MeOD)δ7.91(s,1H),7.31-7.26(m,5H),7.12(d,J＝7.6Hz,1H),6.68(s,1H),6.57(d,J＝7.6Hz,1H),6.41(s,1H),5.30(s,2H),4.89(s,2H),4.47(s,2H),4.32(s,2H),2.47(s,3H),2.38(s,3H)；ESI-MS(m/z)：468.3[M+H] +。 The operation process is the same as in Example 9, except that the aniline in step a is replaced with benzylamine to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-(benzylamine). Amino)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000059
-2-Carboxamide, 1 HNMR (400MHz, MeOD) δ7.91 (s, 1H), 7.31-7.26 (m, 5H), 7.12 (d, J = 7.6 Hz, 1H), 6.68 (s, 1H), 6.57(d,J=7.6Hz,1H), 6.41(s, 1H), 5.30(s, 2H), 4.89(s, 2H), 4.47(s, 2H), 4.32(s, 2H), 2.47(s , 3H), 2.38 (s, 3H); ESI-MS (m/z): 468.3 [M+H] + .
实施例12Example 12
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-(苄氧基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000060
-2-甲酰胺(化合物12)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-8-(benzyloxy)-10,11-dihydro-5H-benzo[e]imidazole[1 ,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000060
Preparation of -2-carboxamide (Compound 12)
Figure PCTCN2020137640-appb-000061
Figure PCTCN2020137640-appb-000061
步骤a)：4-(苄氧基)-2-硝基苯甲酸甲酯的制备Step a): Preparation of methyl 4-(benzyloxy)-2-nitrobenzoate
将4-羟基-2-硝基苯甲酸甲酯(5.0g，25.361mmol)、苯甲醇(2.74g，25.361mmol)、三苯基膦(7.32g，27.897mmol)和无水四氢呋喃(100mL)加入反应瓶中，以氩气置换瓶中空气，在0℃条件下缓慢加入偶氮二甲酸二乙酯(4.86g，27.897mmol)，升温至室温搅拌反应过夜，反应结束后，减压浓缩，所得粗品以乙酸乙酯(200mL)溶解，依次用水(100mL)和饱和氯化钠水溶液(80mL×2)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，残余物经硅胶柱层析纯化(洗脱剂：石油醚/乙酸乙酯＝10/1)，得4-(苄氧基)-2-硝基苯甲酸甲酯，收率30.2％， 1H NMR(400MHz,CDCl 3)δ7.79(d,J＝7.6Hz,1H),7.50-7.27(m,5H),7.36(d,J＝7.6Hz,1H),7.15(dd,J 1＝7.6Hz,J 2＝2.0Hz,1H),5.14(s,2H),3.88(s,3H)；ESI-MS(m/z)：288.1[M+H] +。 Methyl 4-hydroxy-2-nitrobenzoate (5.0g, 25.361mmol), benzyl alcohol (2.74g, 25.361mmol), triphenylphosphine (7.32g, 27.897mmol) and anhydrous tetrahydrofuran (100mL) were added In the reaction flask, replace the air in the flask with argon, slowly add diethyl azodicarboxylate (4.86g, 27.897mmol) at 0°C, warm up to room temperature, stir and react overnight. After the reaction is complete, concentrate under reduced pressure to obtain The crude product was dissolved in ethyl acetate (200 mL), washed successively with water (100 mL) and saturated aqueous sodium chloride solution (80 mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography ( Eluent: petroleum ether/ethyl acetate = 10/1) to obtain methyl 4-(benzyloxy)-2-nitrobenzoate with a yield of 30.2%, 1 H NMR (400MHz, CDCl 3 )δ7. 79 (d, J = 7.6 Hz, 1H), 7.50-7.27 (m, 5H), 7.36 (d, J = 7.6 Hz, 1H), 7.15 (dd, J 1 ＝7.6 Hz, J 2 ＝2.0 Hz, 1H ), 5.14 (s, 2H), 3.88 (s, 3H); ESI-MS (m/z): 288.1 [M+H] + .
步骤b-i)：N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-(苄氧基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000062
-2-甲酰胺的制备 Step bi): N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-(benzyloxy)-10,11-dihydro-5H-benzo[e ]Imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000062
Preparation of -2-carboxamide
操作过程同实施例2中的步骤b-i，只是将实施例2中的4-((1H-吡唑-1-基)甲基)-2-硝基苯甲酸甲酯用相应的4-(苄氧基)-2-硝基苯甲酸甲酯替代，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-(苄氧基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000063
-2-甲酰胺， 1H NMR(400MHz,MeOD)δ8.01(s,1H),7.48-7.27(m,5H),7.16(d,J＝7.6Hz,1H),6.58(s,1H),6.56(d,J＝7.6Hz,1H),6.43(s,1H),5.30(s,2H),5.12(s,2H),4.89(s,2H),4.47(s,2H),2.56(s,3H),2.42(s,3H)； ESI-MS(m/z)：469.3[M+H] +。 The operation process is the same as the step bi in Example 2, except that the methyl 4-((1H-pyrazol-1-yl)methyl)-2-nitrobenzoate in Example 2 is used with the corresponding 4-(benzyl) Oxy)-2-nitrobenzoic acid methyl ester is substituted to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-(benzyloxy)-10, 11-Dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000063
-2-Carboxamide, 1 H NMR(400MHz, MeOD)δ8.01(s,1H), 7.48-7.27(m,5H), 7.16(d,J=7.6Hz,1H), 6.58(s,1H) ,6.56(d,J=7.6Hz,1H),6.43(s,1H),5.30(s,2H),5.12(s,2H),4.89(s,2H),4.47(s,2H),2.56( s, 3H), 2.42 (s, 3H); ESI-MS (m/z): 469.3 [M+H] + .
实施例13Example 13
N-((1-氨基异喹啉-6-基)甲基)-8-((4-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000064
-2-甲酰胺(化合物13)的制备 N-((1-Aminoisoquinolin-6-yl)methyl)-8-((4-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H- Benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000064
Preparation of -2-carboxamide (Compound 13)
Figure PCTCN2020137640-appb-000065
Figure PCTCN2020137640-appb-000065
操作过程同实施例2，只是分别将步骤a中的1H-吡唑用4-甲基-1H-吡唑替代，将步骤i中的5-(氨甲基)-4,6-二甲基吡啶-2-二胺二盐酸盐用6-(氨甲基)异喹啉-1-胺替代，得N-((1-氨基异喹啉-6-基)甲基)-8-((4-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000066
-2-甲酰胺， 1H NMR(400MHz,DMSO-d 6)δ8.90(brs,1H),8.72(t,J＝6.4Hz,1H),8.47(d,J＝8.4Hz,1H),7.75(s,2H),7.68(d,J＝8.4Hz,1H),7.62(d,J＝7.20Hz,1H),7.45(s,1H),7.25-7.19(m,2H),6.97(d,J＝7.2Hz,1H),6.44-6.39(m,2H),5.35(s,2H),5.06-5.05(m,2H),4.59(brd,J＝5.60Hz,2H),4.44(s,2H),1.98(s,3H)；ESI-MS(m/z)：479.3[M+H] +。 The operation process is the same as in Example 2, except that the 1H-pyrazole in step a is replaced by 4-methyl-1H-pyrazole, and the 5-(aminomethyl)-4,6-dimethyl in step i is replaced by 4-methyl-1H-pyrazole. The pyridine-2-diamine dihydrochloride is replaced with 6-(aminomethyl)isoquinolin-1-amine to obtain N-((1-aminoisoquinolin-6-yl)methyl)-8-( (4-Methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000066
-2-Carboxamide, 1 H NMR (400MHz, DMSO-d 6 )δ8.90(brs,1H), 8.72(t,J=6.4Hz,1H), 8.47(d,J=8.4Hz,1H), 7.75(s,2H),7.68(d,J=8.4Hz,1H),7.62(d,J=7.20Hz,1H),7.45(s,1H),7.25-7.19(m,2H),6.97(d ,J=7.2Hz,1H),6.44-6.39(m,2H),5.35(s,2H),5.06-5.05(m,2H),4.59(brd,J=5.60Hz,2H),4.44(s, 2H), 1.98 (s, 3H); ESI-MS (m/z): 479.3 [M+H] + .
实施例14Example 14
N-((1-氨基异喹啉-6-基)甲基)-8-((4-氟-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000067
-2-甲酰胺(化合物14)的制备 N-((1-Aminoisoquinolin-6-yl)methyl)-8-((4-fluoro-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzene And [e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000067
Preparation of -2-carboxamide (Compound 14)
Figure PCTCN2020137640-appb-000068
Figure PCTCN2020137640-appb-000068
操作过程同实施例2，只是分别将步骤a中的1H-吡唑用4-氟-1H-吡唑替代，将步骤i中的5-(氨甲基)-4,6-二甲基吡啶-2-二胺二盐酸盐用6-(氨甲基)异喹啉-1-胺替代，得N-((1-氨基异喹啉-6-基)甲基)-8-((4-氟-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000069
-2-甲酰胺， 1H NMR(400MHz,DMSO-d 6)δ8.97-8.81(m,2H),8.59-8.47(m,1H),7.96(s,1H),7.91-7.87(m,1H),7.76(s,1H),7.60(s,1H),7.52(d,J＝7.2Hz,1H),7.48-7.41(m,2H),7.23(d,J＝6.8Hz,1H),7.04-6.97(m,1H),6.47-6.40(m,2H),5.36(s,2H),5.07(s,2H),4.58-4.44(m,4H)；ESI-MS(m/z)：483.3[M+H] +。 The operation process is the same as in Example 2, except that the 1H-pyrazole in step a is replaced by 4-fluoro-1H-pyrazole, and the 5-(aminomethyl)-4,6-lutidine in step i is replaced by 4-fluoro-1H-pyrazole. -2-Diamine dihydrochloride is replaced with 6-(aminomethyl)isoquinolin-1-amine to obtain N-((1-aminoisoquinolin-6-yl)methyl)-8-(( 4-Fluoro-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000069
-2-Carboxamide, 1 H NMR(400MHz, DMSO-d 6 )δ8.97-8.81(m,2H), 8.59-8.47(m,1H),7.96(s,1H),7.91-7.87(m, 1H), 7.76 (s, 1H), 7.60 (s, 1H), 7.52 (d, J = 7.2Hz, 1H), 7.48-7.41 (m, 2H), 7.23 (d, J = 6.8Hz, 1H), 7.04-6.97(m,1H),6.47-6.40(m,2H),5.36(s,2H),5.07(s,2H),4.58-4.44(m,4H); ESI-MS(m/z): 483.3[M+H] + .
实施例15Example 15
N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-((4-氟-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000070
-2-甲酰胺(化合物15)的制备 N-((1-amino-7-methoxyisoquinolin-6-yl)methyl)-8-((4-fluoro-1H-pyrazol-1-yl)methyl)-10,11- Dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000070
Preparation of -2-carboxamide (Compound 15)
Figure PCTCN2020137640-appb-000071
Figure PCTCN2020137640-appb-000071
步骤a)：7-甲氧基-1-((4-甲氧基苄基)氨基)异喹啉-6-甲腈的制备Step a): Preparation of 7-methoxy-1-((4-methoxybenzyl)amino)isoquinoline-6-carbonitrile
将1-氯-7-甲氧基异喹啉-6-甲腈(900mg，4.116mmol)、4-甲氧基苄胺(2.26g，16.464mmol)和二氧六环(1mL)加入反应瓶中，氮气保护下升温至120℃反应12h，反应结束后，冷却至室温，加入甲基叔丁基醚(15mL)搅拌30min，过滤，滤饼减压真空干燥，得7-甲氧基-1-((4-甲氧基苄基)氨基)异喹啉-6-甲腈， 1H NMR(400MHz,CDCl 3)δ8.07-8.03(m,2H),7.41(d,J＝8.8Hz,2H),7.01(s,1H),6.98-6.94(m,3H),5.20(brs,1H),4.77(d,J＝5.2Hz,2H),4.02(s,3H),3.85(s,3H)；ESI-MS(m/z)：320.1[M+H] +。 Add 1-chloro-7-methoxyisoquinoline-6-carbonitrile (900mg, 4.116mmol), 4-methoxybenzylamine (2.26g, 16.464mmol) and dioxane (1mL) into the reaction flask Under the protection of nitrogen, the temperature was raised to 120°C and reacted for 12h. After the reaction, it was cooled to room temperature, methyl tert-butyl ether (15mL) was added and stirred for 30min, filtered, and the filter cake was dried under reduced pressure and vacuum to obtain 7-methoxy-1 -((4-Methoxybenzyl)amino)isoquinoline-6-carbonitrile, 1 H NMR(400MHz, CDCl 3 )δ8.07-8.03(m,2H),7.41(d,J=8.8Hz ,2H),7.01(s,1H),6.98-6.94(m,3H),5.20(brs,1H),4.77(d,J=5.2Hz,2H),4.02(s,3H),3.85(s, 3H); ESI-MS (m/z): 320.1 [M+H] + .
步骤b)：6-(氨甲基)-7-甲氧基-N-(4-甲氧基苄基)异喹啉-1-胺的制备Step b): Preparation of 6-(aminomethyl)-7-methoxy-N-(4-methoxybenzyl)isoquinolin-1-amine
将7-甲氧基-1-((4-甲氧基苄基)氨基)异喹啉-6-甲腈(1.40g，4.384mmol)和甲醇(40mL)加入反应瓶中，搅拌溶解，加入氨水(40mL)和Raney-Ni(700mg)，反应液在30℃通氢气(50psi)反应12h，反应结束后，过滤，滤饼适量甲醇洗涤，合并滤液，减压浓缩，得6-(氨甲基)-7-甲氧基-N-(4-甲氧基苄基)异喹啉-1-胺，ESI-MS(m/z)：324.2[M+H] +。 Add 7-methoxy-1-((4-methoxybenzyl)amino)isoquinoline-6-carbonitrile (1.40g, 4.384mmol) and methanol (40mL) into the reaction flask, stir to dissolve, add Ammonia (40mL) and Raney-Ni (700mg), the reaction solution was reacted with hydrogen (50psi) at 30°C for 12h. After the reaction, it was filtered, the filter cake was washed with an appropriate amount of methanol, the filtrates were combined, and concentrated under reduced pressure to obtain 6-(aminomethyl) Yl)-7-methoxy-N-(4-methoxybenzyl)isoquinolin-1-amine, ESI-MS (m/z): 324.2 [M+H] + .
步骤c)：6-(氨甲基)-7-甲氧基异喹啉-1-胺的制备Step c): Preparation of 6-(aminomethyl)-7-methoxyisoquinolin-1-amine
将6-(氨甲基)-7-甲氧基-N-(4-甲氧基苄基)异喹啉-1-胺(200mg，0.618mmol)和三氟乙酸(3mL)加入反应瓶中，氮气保护下升温至50℃反应5h，反应结束后，冷却至室温，加水搅拌30min，以二氯甲烷(15mL×3)萃取，水层加入浓盐酸(3mL)，减压浓缩除去大部分水，再加入适量甲苯共沸除水，得6-(氨甲基)-7-甲氧基异喹啉-1-胺，收率78.1％， 1H NMR(400MHz,DMSO-d 6)δ9.18(brs,2H),8.50(brs,2H),8.19(s,1H),7.98(s,1H),7.63(d,J＝6.8Hz,1H),7.15(d,J＝6.8Hz,1H),4.15(s,2H),4.02(s,3H)；ESI-MS(m/z)：204.1[M+H] +。 Add 6-(aminomethyl)-7-methoxy-N-(4-methoxybenzyl)isoquinolin-1-amine (200mg, 0.618mmol) and trifluoroacetic acid (3mL) into the reaction flask Under the protection of nitrogen, the temperature was raised to 50°C and reacted for 5h. After the reaction, it was cooled to room temperature, water was added and stirred for 30 minutes, and extracted with dichloromethane (15mL×3). The aqueous layer was added with concentrated hydrochloric acid (3mL), concentrated under reduced pressure to remove most of the water , Then add an appropriate amount of toluene to azeotropically remove water to obtain 6-(aminomethyl)-7-methoxyisoquinolin-1-amine, the yield is 78.1%, 1 H NMR (400MHz, DMSO-d 6 ) δ9. 18 (brs, 2H), 8.50 (brs, 2H), 8.19 (s, 1H), 7.98 (s, 1H), 7.63 (d, J = 6.8 Hz, 1H), 7.15 (d, J = 6.8 Hz, 1H ), 4.15 (s, 2H), 4.02 (s, 3H); ESI-MS (m/z): 204.1 [M+H] + .
步骤d)：N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-((4-氟-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000072
-2-甲酰胺的制备 Step d): N-((1-amino-7-methoxyisoquinolin-6-yl)methyl)-8-((4-fluoro-1H-pyrazol-1-yl)methyl)- 10,11-Dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000072
Preparation of -2-carboxamide
以8-((4-氟-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000073
-2-羧酸和6-(氨甲基)-7-甲氧基异喹啉-1-胺为原料，操作过程同实施例2中的步骤i，得N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-((4-氟-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000074
-2-甲酰胺， 1H NMR(400MHz,DMSO-d 6)δ8.92-8.73(m,2H),8.61-8.52(m,1H),7.96(s,1H),7.91-7.87(m,1H),7.78(s,1H),7.59(s,1H),7.52(d,J＝6.8Hz,1H),7.48-7.45(m,1H),7.23(d,J＝6.8Hz,1H),7.04-6.97(m,1H),6.47-6.40(m,2H),5.36(s,2H),5.07(s,2H),4.56-4.44(m,4H),4.06-3.96(m,3H)；ESI-MS(m/z)：513.2[M+H] +。 Take 8-((4-fluoro-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4] two Aza
Figure PCTCN2020137640-appb-000073
-2-carboxylic acid and 6-(aminomethyl)-7-methoxyisoquinolin-1-amine as raw materials, the operation process is the same as the step i in Example 2, to obtain N-((1-amino-7 -Methoxyisoquinolin-6-yl)methyl)-8-((4-fluoro-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo(e ]Imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000074
-2-Carboxamide, 1 H NMR(400MHz, DMSO-d 6 )δ8.92-8.73(m,2H),8.61-8.52(m,1H),7.96(s,1H),7.91-7.87(m, 1H), 7.78 (s, 1H), 7.59 (s, 1H), 7.52 (d, J = 6.8 Hz, 1H), 7.48-7.45 (m, 1H), 7.23 (d, J = 6.8 Hz, 1H), 7.04-6.97(m,1H),6.47-6.40(m,2H),5.36(s,2H),5.07(s,2H),4.56-4.44(m,4H),4.06-3.96(m,3H); ESI-MS (m/z): 513.2 [M+H] + .
实施例16Example 16
N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-((2-氧代吡啶-1(2H)-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000075
-2-甲酰胺(化合物16)的制备 N-((1-amino-7-methoxyisoquinolin-6-yl)methyl)-8-((2-oxopyridine-1(2H)-yl)methyl)-10,11- Dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000075
Preparation of -2-carboxamide (Compound 16)
Figure PCTCN2020137640-appb-000076
Figure PCTCN2020137640-appb-000076
以8-((2-氧代吡啶-1(2H)-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000077
-2-羧酸和6-(氨甲基)-7-甲氧基异喹啉-1-胺为原料，操作过程同实施例2中的步骤i，得N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-((2-氧代吡啶-1(2H)-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000078
-2-甲酰胺； 1H NMR(400MHz,DMSO-d 6)δ8.88-8.71(m,2H),8.57-8.52(m,1H),8.01-7.94(m,1H),7.91-7.87(m,1H),7.78(s,1H),7.59-7.52(m,2H),7.52(d,J＝6.8Hz,1H),7.23(d,J＝6.8Hz,1H),7.04-6.97(m,1H),6.60(d,J＝7.6Hz,1H),6.47-6.40(m,2H),6.39(d,J＝7.6Hz,1H),5.31(s,2H),4.91(s,2H),4.41(s,2H),4.29(d,J＝5.2Hz,2H),4.00(s,3H)；ESI-MS(m/z)：522.3[M+H] +。 Take 8-((2-oxopyridine-1(2H)-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4] two Aza
Figure PCTCN2020137640-appb-000077
-2-carboxylic acid and 6-(aminomethyl)-7-methoxyisoquinolin-1-amine as raw materials, the operation process is the same as the step i in Example 2, to obtain N-((1-amino-7 -Methoxyisoquinolin-6-yl)methyl)-8-((2-oxopyridin-1(2H)-yl)methyl)-10,11-dihydro-5H-benzo(e ]Imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000078
-2-carboxamide; 1 H NMR (400MHz, DMSO-d 6 ) δ 8.88-8.71 (m, 2H), 8.57-8.52 (m, 1H), 8.01-7.94 (m, 1H), 7.91-7.87 ( m,1H),7.78(s,1H),7.59-7.52(m,2H),7.52(d,J=6.8Hz,1H),7.23(d,J=6.8Hz,1H),7.04-6.97(m ,1H),6.60(d,J=7.6Hz,1H),6.47-6.40(m,2H),6.39(d,J=7.6Hz,1H),5.31(s,2H),4.91(s,2H) , 4.41 (s, 2H), 4.29 (d, J = 5.2 Hz, 2H), 4.00 (s, 3H); ESI-MS (m/z): 522.3 [M+H] + .
实施例17Example 17
N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-((2-氧代噁唑烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000079
-2-甲酰胺(化合物17)的制备 N-((1-Amino-7-methoxyisoquinolin-6-yl)methyl)-8-((2-oxazolidin-3-yl)methyl)-10,11-bis Hydrogen-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000079
Preparation of -2-carboxamide (Compound 17)
Figure PCTCN2020137640-appb-000080
Figure PCTCN2020137640-appb-000080
以8-((2-氧代噁唑烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000081
-2-羧酸和6-(氨甲基)-7-甲氧基异喹啉-1-胺为原料，操作过程同实施例2中的步骤i，得N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-((2-氧代噁唑烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000082
-2-甲酰胺； 1H NMR(400MHz,MeOD)δ8.19(s,1H),8.00(s,1H),7.98(s,1H),7.63(d,J＝6.8Hz,1H),7.38(d,J＝7.6Hz,1H),7.15(d,J＝6.8Hz,1H),7.04-7.00(m,2H),5.36(s,2H),5.11(s,2H),4.56-4.49(m,4H),4.15(s,2H),4.03(dd,J 1＝7.6,J 2＝6.4Hz,2H),4.02(s,3H)；ESI-MS(m/z)：514.2[M+H] +。 As 8-((2-oxooxazolidin-3-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine miscellaneous
Figure PCTCN2020137640-appb-000081
-2-carboxylic acid and 6-(aminomethyl)-7-methoxyisoquinolin-1-amine as raw materials, the operation process is the same as the step i in Example 2, to obtain N-((1-amino-7 -Methoxyisoquinolin-6-yl)methyl)-8-((2-oxazolidin-3-yl)methyl)-10,11-dihydro-5H-benzo[e] Imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000082
-2-Carboxamide; 1 H NMR (400MHz, MeOD) δ 8.19 (s, 1H), 8.00 (s, 1H), 7.98 (s, 1H), 7.63 (d, J = 6.8 Hz, 1H), 7.38 (d,J=7.6Hz,1H),7.15(d,J=6.8Hz,1H),7.04-7.00(m,2H),5.36(s,2H),5.11(s,2H),4.56-4.49( m,4H),4.15(s,2H),4.03(dd,J 1 =7.6,J 2 =6.4Hz,2H),4.02(s,3H); ESI-MS(m/z): 514.2[M+ H] + .
实施例18Example 18
N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000083
-2-甲酰胺(化合物18)的制备 N-((1-amino-7-methoxyisoquinolin-6-yl)methyl)-8-((2-oxo-3-azabicyclo[3.1.0]hexane-3-yl )Methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000083
Preparation of -2-carboxamide (Compound 18)
Figure PCTCN2020137640-appb-000084
Figure PCTCN2020137640-appb-000084
以8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000085
-2-羧酸和6-(氨甲基)-7-甲氧基异喹啉-1-胺为原料，操作过程同实施例2中的步骤i，得N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000086
-2-甲酰胺； 1H NMR(400MHz,MeOD)δ8.19(s,1H),8.16-7.93(m,1H),7.98(s,1H),7.60(d,J＝6.8Hz,1H),7.38(d,J＝7.6Hz,1H),7.15(d,J＝6.8 Hz,1H),7.04-6.84(m,2H),5.36(s,2H),5.11(s,2H),4.50(s,2H),4.39-4.36(m,2H),3.50-3.28(m,1H),3.14-3.0(m,1H),1.99-1.78(m,2H),1.10-1.03(m,1H),0.58-0.51(m,1H)；ESI-MS(m/z)：524.3[M+H] +。 Take 8-((2-oxo-3-azabicyclo[3.1.0]hexane-3-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2 -a][1,4]diazepine
Figure PCTCN2020137640-appb-000085
-2-carboxylic acid and 6-(aminomethyl)-7-methoxyisoquinolin-1-amine as raw materials, the operation process is the same as the step i in Example 2, to obtain N-((1-amino-7 -Methoxyisoquinolin-6-yl)methyl)-8-((2-oxo-3-azabicyclo[3.1.0]hexane-3-yl)methyl)-10,11- Dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000086
-2-Carboxamide; 1 H NMR (400MHz, MeOD) δ 8.19 (s, 1H), 8.16-7.93 (m, 1H), 7.98 (s, 1H), 7.60 (d, J = 6.8 Hz, 1H) ,7.38(d,J=7.6Hz,1H),7.15(d,J=6.8 Hz,1H),7.04-6.84(m,2H),5.36(s,2H),5.11(s,2H),4.50( s, 2H), 4.39-4.36 (m, 2H), 3.50-3.28 (m, 1H), 3.14-3.0 (m, 1H), 1.99-1.78 (m, 2H), 1.10-1.03 (m, 1H), 0.58-0.51 (m, 1H); ESI-MS (m/z): 524.3 [M+H] + .
实施例19Example 19
N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-(吡咯烷-1-基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000087
-2-甲酰胺(化合物19)的制备 N-((1-amino-7-methoxyisoquinolin-6-yl)methyl)-8-(pyrrolidin-1-yl)-10,11-dihydro-5H-benzo[e] Imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000087
Preparation of -2-carboxamide (Compound 19)
Figure PCTCN2020137640-appb-000088
Figure PCTCN2020137640-appb-000088
以N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-(吡咯烷-1-基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000089
-2-羧酸和6-(氨甲基)-7-甲氧基异喹啉-1-胺为原料，操作过程同实施例2中的步骤i，得N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-(吡咯烷-1-基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000090
-2-甲酰胺；ESI-MS(m/z)：484.3[M+H] +。 With N-((1-amino-7-methoxyisoquinolin-6-yl)methyl)-8-(pyrrolidin-1-yl)-10,11-dihydro-5H-benzo(e ]Imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000089
-2-carboxylic acid and 6-(aminomethyl)-7-methoxyisoquinolin-1-amine as raw materials, the operation process is the same as the step i in Example 2, to obtain N-((1-amino-7 -Methoxyisoquinolin-6-yl)methyl)-8-(pyrrolidin-1-yl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][ 1,4] Diaza
Figure PCTCN2020137640-appb-000090
-2-carboxamide; ESI-MS (m/z): 484.3 [M+H] + .
实施例20Example 20
N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-(苯胺基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000091
-2-甲酰胺(化合物20)的制备 N-((1-amino-7-methoxyisoquinolin-6-yl)methyl)-8-(anilino)-10,11-dihydro-5H-benzo[e]imidazole[1, 2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000091
Preparation of -2-carboxamide (Compound 20)
Figure PCTCN2020137640-appb-000092
Figure PCTCN2020137640-appb-000092
以N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-(苯胺基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000093
-2-羧酸和6-(氨甲基)-7-甲氧基异喹啉-1-胺为原料，操作过程同实施例2中的步骤i，得N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-(苯胺基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000094
-2-甲酰胺；ESI-MS(m/z)：506.3[M+H] +。 Take N-((1-amino-7-methoxyisoquinolin-6-yl)methyl)-8-(anilino)-10,11-dihydro-5H-benzo[e]imidazole [1 ,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000093
-2-carboxylic acid and 6-(aminomethyl)-7-methoxyisoquinolin-1-amine as raw materials, the operation process is the same as the step i in Example 2, to obtain N-((1-amino-7 -Methoxyisoquinolin-6-yl)methyl)-8-(anilino)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4] Diaza
Figure PCTCN2020137640-appb-000094
-2-carboxamide; ESI-MS (m/z): 506.3 [M+H] + .
实施例21Example 21
N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-苯氧基-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000095
-2-甲酰胺(化合物21)的制备 N-((1-Amino-7-methoxyisoquinolin-6-yl)methyl)-8-phenoxy-10,11-dihydro-5H-benzo[e]imidazole[1,2 -a][1,4]diazepine
Figure PCTCN2020137640-appb-000095
Preparation of -2-carboxamide (Compound 21)
Figure PCTCN2020137640-appb-000096
Figure PCTCN2020137640-appb-000096
以N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-苯氧基-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000097
-2-羧酸和6-(氨甲基)-7-甲氧基异喹啉-1-胺为原料，操作过程同实施例2中的步骤i，得N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-苯氧基-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000098
-2-甲酰胺；ESI-MS(m/z)：507.3[M+H] +。 With N-((1-amino-7-methoxyisoquinolin-6-yl)methyl)-8-phenoxy-10,11-dihydro-5H-benzo[e]imidazole [1, 2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000097
-2-carboxylic acid and 6-(aminomethyl)-7-methoxyisoquinolin-1-amine as raw materials, the operation process is the same as the step i in Example 2, to obtain N-((1-amino-7 -Methoxyisoquinolin-6-yl)methyl)-8-phenoxy-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]two Aza
Figure PCTCN2020137640-appb-000098
-2-carboxamide; ESI-MS (m/z): 507.3 [M+H] + .
实施例22Example 22
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((5-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H- 苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000099
-2-甲酰胺(化合物22)的制备 N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-11-oxy Generation-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000099
Preparation of -2-carboxamide (Compound 22)
Figure PCTCN2020137640-appb-000100
Figure PCTCN2020137640-appb-000100
步骤a)：8-((5-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000101
-2-羧酸的制备 Step a): 8-((5-methyl-1H-pyrazol-1-yl)methyl)-11-oxo-10,11-dihydro-5H-benzo[e]imidazole[1,2 -a][1,4]diazepine
Figure PCTCN2020137640-appb-000101
Preparation of -2-carboxylic acid
将8-((5-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000102
-2-甲酸叔丁酯(350mg，0.890mmol)和5N盐酸(0.71mL)加入反应瓶中，升温至45℃反应6h，反应结束后，减压蒸除溶剂，所得粗品直接用于下一步反应，收率86.6％；ESI-MS(m/z)：338.1[M+H] +。 8-((5-methyl-1H-pyrazol-1-yl)methyl)-11-oxo-10,11-dihydro-5H-benzo[e]imidazole[1,2-a] [1,4] Diaza
Figure PCTCN2020137640-appb-000102
Tert-Butyl-2-carboxylate (350mg, 0.890mmol) and 5N hydrochloric acid (0.71mL) were added to the reaction flask, heated to 45℃ and reacted for 6h. After the reaction, the solvent was evaporated under reduced pressure, and the crude product obtained was directly used in the next reaction , The yield is 86.6%; ESI-MS (m/z): 338.1 [M+H] + .
步骤b)：N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((5-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000103
-2-甲酰胺的制备 Step b): N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl) -11-oxo-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000103
Preparation of -2-carboxamide
将N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((5-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000104
-2-羧酸(200mg，0.593mmol)、5-(氨甲基)-4,6-二甲基吡啶-2-二胺(95.6mg，0.652mmol)、HATU(338mg，0.890mmol)、三乙胺(0.17mL，1.186mmol)和DMF(4mL)加入反应瓶中，室温搅拌反应5h，反应结束后，减压浓缩，残余物经制备HPLC纯化，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((5-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000105
-2-甲酰胺，收率58.5％， 1H NMR(400MHz,MeOD)δ8.01(s,1H),7.95(d,J＝2.0Hz,1H),7.55(d,J＝8.0Hz,1H),7.07(d,J＝8.0Hz,1H),7.01(s,1H),6.72(s,1H),6.52(s,1H),5.55(s,2H),5.42(s,2H),4.51(s,2H),2.61(s,3H),2.49(s,3H),2.42(s,3H)；ESI-MS(m/z)：471.3[M+H] +。 The N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-11- Oxo-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000104
-2-carboxylic acid (200mg, 0.593mmol), 5-(aminomethyl)-4,6-lutidine-2-diamine (95.6mg, 0.652mmol), HATU (338mg, 0.890mmol), three Ethylamine (0.17mL, 1.186mmol) and DMF (4mL) were added to the reaction flask, and the reaction was stirred at room temperature for 5h. After the reaction was completed, it was concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain N-((6-amino-2, 4-dimethylpyridin-3-yl)methyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-11-oxo-10,11-dihydro-5H -Benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000105
-2-Carboxamide, yield 58.5%, 1 H NMR (400MHz, MeOD) δ8.01 (s, 1H), 7.95 (d, J = 2.0 Hz, 1H), 7.55 (d, J = 8.0 Hz, 1H ), 7.07 (d, J = 8.0 Hz, 1H), 7.01 (s, 1H), 6.72 (s, 1H), 6.52 (s, 1H), 5.55 (s, 2H), 5.42 (s, 2H), 4.51 (s, 2H), 2.61 (s, 3H), 2.49 (s, 3H), 2.42 (s, 3H); ESI-MS (m/z): 471.3 [M+H] + .
实施例23Example 23
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((4-氟-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000106
-2-甲酰胺(化合物23)的制备 N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((4-fluoro-1H-pyrazol-1-yl)methyl)-11-oxo -10,11-Dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000106
Preparation of -2-carboxamide (Compound 23)
Figure PCTCN2020137640-appb-000107
Figure PCTCN2020137640-appb-000107
以8-((4-氟-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000108
-2-甲酸叔丁酯为原料，其他操作过程同实施例22，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((4-氟-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000109
-2-甲酰胺； 1H NMR(400MHz,MeOD)δ8.09(d,J＝4.0Hz,1H),8.01(s,1H),7.97(d,J＝2.0Hz,1H),7.53(d,J＝4.0Hz,1H),7.46(d,J＝7.8Hz,1H),7.07(d,J＝7.6Hz,1H),7.01(s,1H),5.35(s, 2H),5.07(s,2H),4.56-4.44(m,4H),2.51(s,3H),2.41(s,3H)；ESI-MS(m/z)：475.3[M+H] +。 With 8-((4-fluoro-1H-pyrazol-1-yl)methyl)-11-oxo-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][ 1,4] Diaza
Figure PCTCN2020137640-appb-000108
-2-carboxylic acid tert-butyl ester as the raw material, and other operating procedures were the same as in Example 22 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((4- Fluoro-1H-pyrazol-1-yl)methyl)-11-oxo-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000109
-2-carboxamide; 1 H NMR (400MHz, MeOD) δ 8.09 (d, J = 4.0 Hz, 1H), 8.01 (s, 1H), 7.97 (d, J = 2.0 Hz, 1H), 7.53 (d ,J=4.0Hz,1H),7.46(d,J=7.8Hz,1H),7.07(d,J=7.6Hz,1H),7.01(s,1H),5.35(s, 2H),5.07(s , 2H), 4.56-4.44 (m, 4H), 2.51 (s, 3H), 2.41 (s, 3H); ESI-MS (m/z): 475.3 [M+H] + .
实施例24Example 24
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-((2-氧代吡啶-1(2H)-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000110
-2-甲酰胺(化合物24)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8-((2-oxopyridin-1(2H)-yl)methyl) -10,11-Dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000110
Preparation of -2-carboxamide (Compound 24)
Figure PCTCN2020137640-appb-000111
Figure PCTCN2020137640-appb-000111
以11-氧代-8-((2-氧代吡啶-1(2H)-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000112
-2-甲酸叔丁酯为原料，其他操作过程同实施例22，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-((2-氧代吡啶-1(2H)-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000113
-2-甲酰胺； 1H NMR(400MHz,DMSO-d 6)δ10.06(brs,1H),8.17(m,1H),7.75(s,1H),7.64-7.60(m,1H),7.60(m,2H),7.40(m,1H),6.98(d,J＝7.6Hz,1H),6.59(s,1H),6.44-6.40(m,2H),6.38(d,J＝7.6Hz,1H),6.20(dt,J 1＝1.2,J 2＝6.8Hz,1H),5.31(s,2H),4.41(s,2H),4.29(d,J＝5.2Hz,2H),2.49(s,3H),2.31(s,3H)；ESI-MS(m/z)：484.3[M+H] +。 Take 11-oxo-8-((2-oxopyridine-1(2H)-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][ 1,4] Diaza
Figure PCTCN2020137640-appb-000112
-2-carboxylic acid tert-butyl ester was used as the raw material, and other procedures were the same as in Example 22 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8 -((2-oxopyridine-1(2H)-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000113
-2-Carboxamide; 1 H NMR (400MHz, DMSO-d 6 ) δ 10.06 (brs, 1H), 8.17 (m, 1H), 7.75 (s, 1H), 7.64-7.60 (m, 1H), 7.60 (m,2H),7.40(m,1H),6.98(d,J=7.6Hz,1H),6.59(s,1H),6.44-6.40(m,2H),6.38(d,J=7.6Hz, 1H), 6.20 (dt, J 1 = 1.2, J 2 = 6.8 Hz, 1H), 5.31 (s, 2H), 4.41 (s, 2H), 4.29 (d, J = 5.2 Hz, 2H), 2.49 (s , 3H), 2.31 (s, 3H); ESI-MS (m/z): 484.3 [M+H] + .
实施例25Example 25
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-((2-氧代噁唑烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000114
-2-甲酰胺(化合物25)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8-((2-oxooxazolidin-3-yl)methyl)- 10,11-Dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000114
Preparation of -2-carboxamide (Compound 25)
Figure PCTCN2020137640-appb-000115
Figure PCTCN2020137640-appb-000115
以11-氧代-8-((2-氧代噁唑烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000116
-2-甲酸叔丁酯为原料，其他操作过程同实施例22，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-((2-氧代噁唑烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000117
-2-甲酰胺； 1H NMR(400MHz,MeOD)δ8.01(s,1H),7.50-7.27(m,1H),6.58(s,1H),6.48-6.40(m,2H),5.84(s,1H),5.35(s,2H),5.10(s,2H),4.53(dd,J 1＝7.6,J 2＝6.4Hz,2H),4.49(s,2H),4.31(s,2H),4.13(dd,J 1＝7.6,J 2＝6.4Hz,2H),2.58(s,3H),2.41(s,3H)；ESI-MS(m/z)：476.3[M+H] +。 Take 11-oxo-8-((2-oxooxazolidin-3-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1 ,4] Diaza
Figure PCTCN2020137640-appb-000116
-2-carboxylic acid tert-butyl ester was used as the raw material, and other procedures were the same as in Example 22 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8 -((2-oxazolidin-3-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000117
-2-carboxamide; 1 H NMR (400MHz, MeOD) δ8.01 (s, 1H), 7.50-7.27 (m, 1H), 6.58 (s, 1H), 6.48-6.40 (m, 2H), 5.84 ( s,1H),5.35(s,2H),5.10(s,2H),4.53(dd,J 1 =7.6,J 2 =6.4Hz,2H),4.49(s,2H),4.31(s,2H) , 4.13 (dd, J 1 =7.6, J 2 =6.4 Hz, 2H), 2.58 (s, 3H), 2.41 (s, 3H); ESI-MS (m/z): 476.3 [M+H] + .
实施例26Example 26
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000118
-2-甲酰胺(化合物26)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8-((2-oxo-3-azabicyclo[3.1.0]hexane (Alk-3-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000118
Preparation of -2-carboxamide (Compound 26)
Figure PCTCN2020137640-appb-000119
Figure PCTCN2020137640-appb-000119
11-氧代-8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000120
-2-甲酸叔丁酯为原料，其他操作过程同实施例22，得N-((6-氨基-2,4-二 甲基吡啶-3-基)甲基)-11-氧代-8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000121
-2-甲酰胺； 1H NMR(400MHz,MeOD)δ8.06(s,1H),7.11(d,J＝7.6Hz,1H),6.80(s,1H),6.64(d,J＝7.6Hz,1H),6.43(s,1H),5.32(s,2H),4.99(s,2H),4.51(s,2H),4.39-4.31(m,2H),3.70-3.56(m,1H),3.17-3.12(m,1H),2.52(s,3H),2.40(s,3H),1.99-1.96(m,1H),1.83-1.78(m,1H),1.21-1.16(m,1H),0.57-0.53(m,1H)；ESI-MS(m/z)：486.2[M+H] +。 11-oxo-8-((2-oxo-3-azabicyclo[3.1.0]hexane-3-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole [1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000120
-2-carboxylic acid tert-butyl ester was used as the raw material, and other procedures were the same as in Example 22 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8 -((2-oxo-3-azabicyclo[3.1.0]hexane-3-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a ][1,4]diazepine
Figure PCTCN2020137640-appb-000121
-2-Carboxamide; 1 H NMR(400MHz, MeOD)δ8.06(s,1H), 7.11(d,J=7.6Hz,1H), 6.80(s,1H), 6.64(d,J=7.6Hz ,1H), 6.43(s, 1H), 5.32(s, 2H), 4.99(s, 2H), 4.51(s, 2H), 4.39-4.31(m, 2H), 3.70-3.56(m, 1H), 3.17-3.12 (m, 1H), 2.52 (s, 3H), 2.40 (s, 3H), 1.99-1.96 (m, 1H), 1.83-1.78 (m, 1H), 1.21-1.16 (m, 1H), 0.57-0.53 (m, 1H); ESI-MS (m/z): 486.2 [M+H] + .
实施例27Example 27
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-(吡咯烷-1-基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000122
-2-甲酰胺(化合物27)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8-(pyrrolidin-1-yl)-10,11-dihydro-5H- Benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000122
Preparation of -2-carboxamide (Compound 27)
Figure PCTCN2020137640-appb-000123
Figure PCTCN2020137640-appb-000123
以11-氧代-8-(吡咯烷-1-基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000124
-2-甲酸叔丁酯为原料，其他操作过程同实施例22，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-(吡咯烷-1-基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000125
-2-甲酰胺；ESI-MS(m/z)：446.3[M+H] +。 Take 11-oxo-8-(pyrrolidin-1-yl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000124
-2-carboxylic acid tert-butyl ester was used as the raw material, and other procedures were the same as in Example 22 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8 -(Pyrrolidin-1-yl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000125
-2-carboxamide; ESI-MS (m/z): 446.3 [M+H] + .
实施例28Example 28
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-(苯胺基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000126
-2-甲酰胺(化合物28)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8-(anilino)-10,11-dihydro-5H-benzo(e ]Imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000126
Preparation of -2-carboxamide (Compound 28)
Figure PCTCN2020137640-appb-000127
Figure PCTCN2020137640-appb-000127
以11-氧代-8-(苯胺基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000128
-2-甲酸叔丁酯为原料，其他操作过程同实施例22，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-(吡咯烷-1-基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000129
-2-甲酰胺；ESI-MS(m/z)：468.3[M+H] +。 Take 11-oxo-8-(anilino)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000128
-2-carboxylic acid tert-butyl ester was used as the raw material, and other procedures were the same as in Example 22 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8 -(Pyrrolidin-1-yl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000129
-2-carboxamide; ESI-MS (m/z): 468.3 [M+H] + .
实施例29Example 29
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-苯氧基-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000130
-2-甲酰胺(化合物29)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8-phenoxy-10,11-dihydro-5H-benzo[e] Imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000130
Preparation of -2-carboxamide (Compound 29)
Figure PCTCN2020137640-appb-000131
Figure PCTCN2020137640-appb-000131
以11-氧代-8-苯氧基-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000132
-2-甲酸叔丁酯为原料，其他操作过程同实施例22，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-苯氧基-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000133
-2-甲酰胺；ESI-MS(m/z)：469.2[M+H] +。 Taking 11-oxo-8-phenoxy-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000132
-2-carboxylic acid tert-butyl ester was used as the raw material, and other procedures were the same as in Example 22 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8 -Phenoxy-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000133
-2-carboxamide; ESI-MS (m/z): 469.2 [M+H] + .
实施例30Example 30
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((5-甲基-1H-吡唑-1-基)甲基)-5H-苯并[e]咪唑 [1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000134
-2-甲酰胺(化合物30)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-5H-benzene And [e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000134
Preparation of -2-carboxamide (Compound 30)
Figure PCTCN2020137640-appb-000135
Figure PCTCN2020137640-appb-000135
步骤a)：8-((5-甲基-1H-吡唑-1-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000136
-2-羧酸的制备 Step a): 8-((5-methyl-1H-pyrazol-1-yl)methyl)-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000136
Preparation of -2-carboxylic acid
将8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000137
-2-羧酸(350mg，1.082mmol)、MnO 2(941mg，10.824mmol)和二氯乙烷(10mL)加入反应瓶中，升温至68℃反应5h，反应结束后，过滤，减压浓缩，所得粗品采用制备型HPLC纯化，得8-((5-甲基-1H-吡唑-1-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000138
-2-羧酸，收率57.5％；ESI-MS(m/z)：322.1[M+H] +。 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4] Diaza
Figure PCTCN2020137640-appb-000137
-2-carboxylic acid (350mg, 1.082mmol), MnO 2 (941mg, 10.824mmol) and dichloroethane (10mL) were added to the reaction flask, heated to 68°C and reacted for 5h, after the reaction, filtered, concentrated under reduced pressure, The obtained crude product was purified by preparative HPLC to obtain 8-((5-methyl-1H-pyrazol-1-yl)methyl)-5H-benzo[e]imidazole[1,2-a][1,4 ] Diaza
Figure PCTCN2020137640-appb-000138
-2-carboxylic acid, the yield is 57.5%; ESI-MS (m/z): 322.1 [M+H] + .
步骤b)：N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((5-甲基-1H-吡唑-1-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000139
-2-甲酰胺 Step b): N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl) -5H-Benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000139
-2-carboxamide
以8-((5-甲基-1H-吡唑-1-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000140
-2-羧酸为原料，操作过程同实施例22中的步骤b，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((5-甲基-1H-吡唑-1-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000141
-2-甲酰胺，收率14.5％， 1H NMR(400MHz,DMSO-d 6)δ8.47(s,1H),7.92(s,1H),7.77(t,J＝5.2Hz,1H),7.42-7.31(m,2H),7.16-6.99(m,2H),6.08(d,J＝3.6Hz,2H),5.67-5.57(m,2H),5.33(s,2H),5.19(s,2H),4.30(d,J＝5.2Hz,2H),2.28(s,3H),2.19(s,3H),2.16(s,3H)；ESI-MS(m/z)：455.3[M+H] +。 Take 8-((5-methyl-1H-pyrazol-1-yl)methyl)-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000140
-2-carboxylic acid as the raw material, the operation process is the same as that of step b in Example 22 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((5 -Methyl-1H-pyrazol-1-yl)methyl)-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000141
-2-carboxamide, yield 14.5%, 1 H NMR (400MHz, DMSO-d 6 )δ8.47(s,1H),7.92(s,1H),7.77(t,J=5.2Hz,1H), 7.42-7.31(m,2H),7.16-6.99(m,2H),6.08(d,J=3.6Hz,2H), 5.67-5.57(m,2H),5.33(s,2H), 5.19(s, 2H), 4.30(d,J=5.2Hz,2H), 2.28(s,3H), 2.19(s,3H), 2.16(s,3H); ESI-MS(m/z): 455.3[M+H ] + .
实施例31Example 31
8-((1H-吡唑-1-基)甲基)-N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000142
-2-甲酰胺(化合物31)的制备 8-((1H-pyrazol-1-yl)methyl)-N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-5H-benzo[e]imidazole [1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000142
Preparation of -2-carboxamide (Compound 31)
Figure PCTCN2020137640-appb-000143
Figure PCTCN2020137640-appb-000143
以8-((1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000144
-2-羧酸为原料，其他操作过程同实施例30，得8-((1H-吡唑-1-基)甲基)-N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000145
-2-甲酰胺， 1H NMR(400MHz,DMSO-d 6)δ8.49-8.47(m,2H),7.92(s,1H),7.84(s,1H),7.73-7.58(m,2H),7.49-7.43(m,1H),7.40-7.34(m,1H),7.23-7.16(m,2H),6.59(s,1H),6.26(s,1H),5.37(s,2H),5.20(s,2H),4.29(d,J＝5.2Hz2H),2.49(s,3H),2.33(s,3H)；ESI-MS(m/z)：441.3[M+H] +。 Take 8-((1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000144
-2-carboxylic acid as the raw material, and the other procedures are the same as in Example 30 to obtain 8-((1H-pyrazol-1-yl)methyl)-N-((6-amino-2,4-lutidine) -3-yl)methyl)-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000145
-2-Carboxamide, 1 H NMR(400MHz, DMSO-d 6 )δ8.49-8.47(m,2H),7.92(s,1H),7.84(s,1H),7.73-7.58(m,2H) ,7.49-7.43(m,1H),7.40-7.34(m,1H),7.23-7.16(m,2H),6.59(s,1H),6.26(s,1H),5.37(s,2H),5.20 (s, 2H), 4.29 (d, J=5.2 Hz 2H), 2.49 (s, 3H), 2.33 (s, 3H); ESI-MS (m/z): 441.3 [M+H] + .
实施例32Example 32
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((4-氟-1H-吡唑-1-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000146
-2-甲酰胺(化合物32)的制备 N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((4-fluoro-1H-pyrazol-1-yl)methyl)-5H-benzo [e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000146
Preparation of -2-carboxamide (Compound 32)
Figure PCTCN2020137640-appb-000147
Figure PCTCN2020137640-appb-000147
以8-((4-氟-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000148
-2-羧酸为原料，其他操作过程同实施例30，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((4-氟-1H-吡唑-1-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000149
-2-甲酰胺；ESI-MS(m/z)：459.3[M+H] +。 Take 8-((4-fluoro-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4] two Aza
Figure PCTCN2020137640-appb-000148
-2-carboxylic acid as the raw material, and other operating procedures were the same as in Example 30 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((4-fluoro- 1H-pyrazol-1-yl)methyl)-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000149
-2-carboxamide; ESI-MS (m/z): 459.3 [M+H] + .
实施例33Example 33
N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-((4-氟-1H-吡唑-1-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000150
-2-甲酰胺(化合物33)的制备 N-((1-amino-7-methoxyisoquinolin-6-yl)methyl)-8-((4-fluoro-1H-pyrazol-1-yl)methyl)-5H-benzo [e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000150
Preparation of -2-carboxamide (Compound 33)
Figure PCTCN2020137640-appb-000151
Figure PCTCN2020137640-appb-000151
以8-((4-氟-1H-吡唑-1-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000152
-2-羧酸和6-(氨甲基)-7-甲氧基异喹啉-1-胺为原料，其他操作过程同实施例22中的步骤b，得N-((1-氨基-7-甲氧基异喹啉-6-基)甲基)-8-((4-氟-1H-吡唑-1-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000153
-2-甲酰胺；ESI-MS(m/z)：511.3[M+H] +。 As 8-((4-fluoro-1H-pyrazol-1-yl)methyl)-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000152
-2-carboxylic acid and 6-(aminomethyl)-7-methoxyisoquinolin-1-amine as raw materials, and the other operating procedures are the same as those in step b in Example 22 to obtain N-((1-amino- 7-Methoxyisoquinolin-6-yl)methyl)-8-((4-fluoro-1H-pyrazol-1-yl)methyl)-5H-benzo[e]imidazole[1,2 -a][1,4]diazepine
Figure PCTCN2020137640-appb-000153
-2-carboxamide; ESI-MS (m/z): 511.3 [M+H] + .
实施例34Example 34
N-((1-氨基异喹啉-6-基)甲基)-8-((4-氟-1H-吡唑-1-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000154
-2-甲酰胺(化合物34)的制备 N-((1-Aminoisoquinolin-6-yl)methyl)-8-((4-fluoro-1H-pyrazol-1-yl)methyl)-5H-benzo[e]imidazole[1 ,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000154
Preparation of -2-carboxamide (Compound 34)
Figure PCTCN2020137640-appb-000155
Figure PCTCN2020137640-appb-000155
以8-((4-氟-1H-吡唑-1-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000156
-2-羧酸和6-(氨甲基)异喹啉-1-胺为原料，其他操作过程同实施例22中的步骤b，得N-((1-氨基异喹啉-6-基)甲基)-8-((4-氟-1H-吡唑-1-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000157
-2-甲酰胺；ESI-MS(m/z)：481.2[M+H] +。 As 8-((4-fluoro-1H-pyrazol-1-yl)methyl)-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000156
-2-carboxylic acid and 6-(aminomethyl)isoquinolin-1-amine are used as raw materials, and the other operating procedures are the same as those in step b in Example 22 to obtain N-((1-aminoisoquinolin-6-yl )Methyl)-8-((4-Fluoro-1H-pyrazol-1-yl)methyl)-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000157
-2-carboxamide; ESI-MS (m/z): 481.2 [M+H] + .
实施例35Example 35
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((2-氧代吡啶-1(2H)-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000158
-2-甲酰胺(化合物35)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-8-((2-oxopyridin-1(2H)-yl)methyl)-5H-benzo [e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000158
Preparation of -2-carboxamide (Compound 35)
Figure PCTCN2020137640-appb-000159
Figure PCTCN2020137640-appb-000159
以8-((2-氧代吡啶-1(2H)-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000160
-2-羧酸为原料，其他操作过程同实施例30，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((2-氧代吡啶-1(2H)-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000161
-2-甲酰胺；ESI-MS(m/z)：468.3[M+H] +。 Take 8-((2-oxopyridine-1(2H)-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4] two Aza
Figure PCTCN2020137640-appb-000160
-2-carboxylic acid as the raw material, and other operating procedures were the same as in Example 30 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((2-oxo Pyridine-1(2H)-yl)methyl)-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000161
-2-carboxamide; ESI-MS (m/z): 468.3 [M+H] + .
实施例36Example 36
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((2-氧代噁唑烷-3-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000162
-2-甲酰胺(化合物36)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-8-((2-oxoxazolidin-3-yl)methyl)-5H-benzo[ e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000162
Preparation of -2-carboxamide (Compound 36)
Figure PCTCN2020137640-appb-000163
Figure PCTCN2020137640-appb-000163
以8-((2-氧代噁唑烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000164
-2-羧酸为原料，其他操作过程同实施例30，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((2-氧代噁唑烷-3-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000165
-2-甲酰胺；ESI-MS(m/z)：460.3[M+H] +。 As 8-((2-oxooxazolidin-3-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine miscellaneous
Figure PCTCN2020137640-appb-000164
-2-carboxylic acid as the raw material, and other operating procedures were the same as in Example 30 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((2-oxo (Oxazolidine-3-yl)methyl)-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000165
-2-carboxamide; ESI-MS (m/z): 460.3 [M+H] + .
实施例37Example 37
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000166
-2-甲酰胺(化合物37)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-8-((2-oxo-3-azabicyclo[3.1.0]hexane-3-yl )Methyl)-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000166
Preparation of -2-carboxamide (Compound 37)
Figure PCTCN2020137640-appb-000167
Figure PCTCN2020137640-appb-000167
以8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000168
-2-羧酸为原料，其他操作过程同实施例30，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基)甲基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000169
-2-甲酰胺；ESI-MS(m/z)：470.3[M+H] +。 Take 8-((2-oxo-3-azabicyclo[3.1.0]hexane-3-yl)methyl)-5H-benzo[e]imidazole[1,2-a][1,4 ] Diaza
Figure PCTCN2020137640-appb-000168
-2-carboxylic acid as the raw material, and other operating procedures were the same as in Example 30 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((2-oxo -3-azabicyclo[3.1.0]hexane-3-yl)methyl)-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000169
-2-carboxamide; ESI-MS (m/z): 470.3 [M+H] + .
实施例38Example 38
N-((6-氨基-2,4-二甲氨基吡啶-3-基)甲基)-8-(吡咯烷-1-基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000170
-2-甲酰胺(化合物38)的制备 N-((6-Amino-2,4-dimethylaminopyridin-3-yl)methyl)-8-(pyrrolidin-1-yl)-5H-benzo[e]imidazole[1,2-a ][1,4]diazepine
Figure PCTCN2020137640-appb-000170
Preparation of -2-carboxamide (Compound 38)
Figure PCTCN2020137640-appb-000171
Figure PCTCN2020137640-appb-000171
以8-(吡咯烷-1-基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000172
-2-羧酸为原料，其他操作过程同实施例30，得N-((6-氨基-2,4-二甲氨基吡啶-3-基)甲基)-8-(吡咯烷-1-基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000173
-2-甲酰胺；ESI-MS(m/z)：430.3[M+H] +。 Take 8-(pyrrolidin-1-yl)-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000172
-2-carboxylic acid as the raw material, and other operating procedures were the same as in Example 30 to obtain N-((6-amino-2,4-dimethylaminopyridin-3-yl)methyl)-8-(pyrrolidine-1- Yl)-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000173
-2-carboxamide; ESI-MS (m/z): 430.3 [M+H] + .
实施例39Example 39
N-((6-氨基-2,4-二甲氨基吡啶-3-基)甲基)-8-(苯胺基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000174
-2-甲酰胺(化合物39)的制备 N-((6-Amino-2,4-dimethylaminopyridin-3-yl)methyl)-8-(anilino)-5H-benzo[e]imidazole[1,2-a][1, 4] Diaza
Figure PCTCN2020137640-appb-000174
Preparation of -2-carboxamide (Compound 39)
Figure PCTCN2020137640-appb-000175
Figure PCTCN2020137640-appb-000175
以8-(苯胺基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000176
-2-羧酸为原料，其他操作过程同实施例 30，得N-((6-氨基-2,4-二甲氨基吡啶-3-基)甲基)-8-(苯胺基)-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000177
-2-甲酰胺；ESI-MS(m/z)：452.3[M+H] +。 Take 8-(anilino)-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000176
-2-carboxylic acid as the raw material, and other operating procedures are the same as in Example 30 to obtain N-((6-amino-2,4-dimethylaminopyridin-3-yl)methyl)-8-(anilino)-5H -Benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000177
-2-carboxamide; ESI-MS (m/z): 452.3 [M+H] + .
实施例40Example 40
N-((6-氨基-2,4-二甲氨基吡啶-3-基)甲基)-8-苯氧基-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000178
-2-甲酰胺(化合物40)的制备 N-((6-Amino-2,4-dimethylaminopyridin-3-yl)methyl)-8-phenoxy-5H-benzo[e]imidazole[1,2-a][1,4 ] Diaza
Figure PCTCN2020137640-appb-000178
Preparation of -2-carboxamide (Compound 40)
Figure PCTCN2020137640-appb-000179
Figure PCTCN2020137640-appb-000179
以8-苯氧基-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000180
-2-羧酸为原料，其他操作过程同实施例30，得N-((6-氨基-2,4-二甲氨基吡啶-3-基)甲基)-8-苯氧基-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000181
-2-甲酰胺；ESI-MS(m/z)：453.3[M+H] +。 With 8-phenoxy-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000180
-2-carboxylic acid was used as the raw material, and the other operating procedures were the same as those in Example 30 to obtain N-((6-amino-2,4-dimethylaminopyridin-3-yl)methyl)-8-phenoxy-5H- Benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000181
-2-carboxamide; ESI-MS (m/z): 453.3 [M+H] + .
实施例41Example 41
N-(4-(氨甲基)苄基)-8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000182
-2-甲酰胺(化合物41)的制备 N-(4-(aminomethyl)benzyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e] Imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000182
Preparation of -2-carboxamide (Compound 41)
Figure PCTCN2020137640-appb-000183
Figure PCTCN2020137640-appb-000183
步骤a)：(4-((8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000184
-2-甲酰胺)甲基)苄基)氨基甲酸叔丁酯的制备 Step a): (4-((8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2 -a][1,4]diazepine
Figure PCTCN2020137640-appb-000184
Preparation of 2-carboxamide) methyl) benzyl) tert-butyl carbamate
将8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000185
-2-羧酸(100mg，0.309mmol)、(4-(氨甲基)苄基)氨基甲酸叔丁酯(110mg，0.464mmol)、三乙胺(78mg，0.773mmol)、HBTU(176mg，0.464mmol)和DMF(3mL)加入反应瓶中，室温搅拌反应1h，反应结束后，反应液加水稀释，以乙酸乙酯(30mL)萃取，合并有机层，饱和氯化钠水溶液(10mL×2)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，得白色固体，无需纯化直接用于下一步反应；ESI-MS(m/z)：542.2[M+H] +。 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4] Diaza
Figure PCTCN2020137640-appb-000185
-2-carboxylic acid (100mg, 0.309mmol), (4-(aminomethyl)benzyl) tert-butyl carbamate (110mg, 0.464mmol), triethylamine (78mg, 0.773mmol), HBTU (176mg, 0.464 mmol) and DMF (3mL) were added to the reaction flask, and the reaction was stirred at room temperature for 1h. After the reaction, the reaction solution was diluted with water and extracted with ethyl acetate (30mL). The organic layers were combined and washed with saturated sodium chloride aqueous solution (10mL×2) , Dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a white solid, which was directly used in the next reaction without purification; ESI-MS (m/z): 542.2 [M+H] + .
步骤b)：N-(4-(氨甲基)苄基)-8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000186
-2-甲酰胺 Step b): N-(4-(aminomethyl)benzyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzene And [e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000186
-2-carboxamide
将(4-((8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000187
-2-甲酰胺)甲基)苄基)氨基甲酸叔丁酯(180mg，0.332mmol)和盐酸(5mL,6N)加入反应瓶中，室温搅拌反应1h，反应结束后，减压浓缩，所得粗品经制备型HPLC纯化，得N-(4-(氨甲基)苄基)-8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000188
-2-甲酰胺，收率22.5％， 1H NMR(400MHz,MeOD)δ7.67(s,1H),7.38(d,J＝1.6Hz,1H),7.33-7.25(m,4H),7.06(d,J＝7.6Hz,1H),6.41-6.38(m,2H),6.10(d,J＝1.6Hz,1H),5.28(s,2H),5.18(s,2H),4.51(s,2H),4.44(s,2H),3.79(d,J＝2.8Hz,2H),2.20(s,3H)；ESI-MS(m/z)： 442.3[M+H] +。 Add (4-((8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a] [1,4] Diaza
Figure PCTCN2020137640-appb-000187
2-Carboxamide)methyl)benzyl)carbamic acid tert-butyl ester (180mg, 0.332mmol) and hydrochloric acid (5mL, 6N) were added to the reaction flask, and the reaction was stirred at room temperature for 1h. After the reaction, it was concentrated under reduced pressure to obtain the crude product. Purified by preparative HPLC, N-(4-(aminomethyl)benzyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro- 5H-Benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000188
-2-carboxamide, yield 22.5%, 1 H NMR (400MHz, MeOD) δ 7.67 (s, 1H), 7.38 (d, J = 1.6 Hz, 1H), 7.33-7.25 (m, 4H), 7.06 (d,J=7.6Hz,1H),6.41-6.38(m,2H),6.10(d,J=1.6Hz,1H),5.28(s,2H),5.18(s,2H),4.51(s, 2H), 4.44 (s, 2H), 3.79 (d, J=2.8 Hz, 2H), 2.20 (s, 3H); ESI-MS (m/z): 442.3 [M+H] + .
实施例42Example 42
N-(4-(1-氨基环丙基)苄基)-8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000189
-2-甲酰胺(化合物42)的制备 N-(4-(1-Aminocyclopropyl)benzyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo [e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000189
Preparation of -2-carboxamide (Compound 42)
Figure PCTCN2020137640-appb-000190
Figure PCTCN2020137640-appb-000190
以8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000191
-2-羧酸和1-(4-(氨甲基)苯基)环丙基-1-胺为原料，其他操作过程同实施例22中的步骤b，得N-(4-(1-氨基环丙基)苄基)-8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000192
-2-甲酰胺， 1H NMR(400MHz,MeOD)δ7.79(s,1H),7.42(d,J＝1.6Hz,1H),7.33-7.27(m,4H),7.06(d,J＝7.6Hz,1H),6.41-6.38(m,2H),6.10(m,1H),5.28(s,2H),5.11(s,2H),4.51(s,2H),3.79(d,J＝2.8Hz,2H),2.21(s,3H),1.22(m,2H),1.10(m,2H)；ESI-MS(m/z)：468.3[M+H] +。 Take 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4] Diaza
Figure PCTCN2020137640-appb-000191
-2-carboxylic acid and 1-(4-(aminomethyl)phenyl)cyclopropyl-1-amine as raw materials, and other operating procedures are the same as those in step b in Example 22 to obtain N-(4-(1- Aminocyclopropyl)benzyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2 -a][1,4]diazepine
Figure PCTCN2020137640-appb-000192
-2-Carboxamide, 1 H NMR (400MHz, MeOD) δ7.79 (s, 1H), 7.42 (d, J = 1.6 Hz, 1H), 7.33-7.27 (m, 4H), 7.06 (d, J = 7.6Hz, 1H), 6.41-6.38 (m, 2H), 6.10 (m, 1H), 5.28 (s, 2H), 5.11 (s, 2H), 4.51 (s, 2H), 3.79 (d, J = 2.8 Hz, 2H), 2.21 (s, 3H), 1.22 (m, 2H), 1.10 (m, 2H); ESI-MS (m/z): 468.3 [M+H] + .
实施例43Example 43
N-(4-(氨甲基)苯基)-8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000193
-2-甲酰胺(化合物43)的制备 N-(4-(Aminomethyl)phenyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e] Imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000193
Preparation of -2-carboxamide (Compound 43)
Figure PCTCN2020137640-appb-000194
Figure PCTCN2020137640-appb-000194
操作过程同实施例41，只是将(4-(氨甲基)苄基)氨基甲酸叔丁酯用(4-氨基苄基)氨基甲酸叔丁酯替代，得N-(4-(氨甲基)苯基)-8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000195
-2-甲酰胺， 1H NMR(400MHz,MeOD)δ7.68(s,1H),7.58(d,J＝7.6Hz,2H),7.38(d,J＝7.6Hz,2H),7.32(d,J＝2.0Hz,1H),7.06(d,J＝7.6Hz,1H),6.41-6.38(m,2H),6.06(d,J＝2.0Hz,1H),5.31(s,2H),5.18(s,2H),4.51(s,2H),4.37(s,2H),2.21(s,3H)；ESI-MS(m/z)：428.3[M+H] +。 The procedure is the same as in Example 41, except that tert-butyl (4-(aminomethyl)benzyl)carbamate is replaced with tert-butyl (4-aminobenzyl)carbamate to obtain N-(4-(aminomethyl) )Phenyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][ 1,4] Diaza
Figure PCTCN2020137640-appb-000195
-2-Carboxamide, 1 H NMR (400MHz, MeOD) δ7.68 (s, 1H), 7.58 (d, J = 7.6 Hz, 2H), 7.38 (d, J = 7.6 Hz, 2H), 7.32 (d ,J=2.0Hz,1H),7.06(d,J=7.6Hz,1H),6.41-6.38(m,2H),6.06(d,J=2.0Hz,1H),5.31(s,2H),5.18 (s, 2H), 4.51 (s, 2H), 4.37 (s, 2H), 2.21 (s, 3H); ESI-MS (m/z): 428.3 [M+H] + .
实施例44Example 44
N-((1H-吡咯[2,3-b]吡啶-5-基)甲基)-8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000196
-2-甲酰胺(化合物44)的制备 N-((1H-pyrrole[2,3-b]pyridin-5-yl)methyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11- Dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000196
Preparation of -2-carboxamide (Compound 44)
Figure PCTCN2020137640-appb-000197
Figure PCTCN2020137640-appb-000197
以8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000198
-2-羧酸和(1H-吡咯[2,3-b]吡啶-5-基)甲胺为原料，操作过程同实施22中的步骤b，得N-((1H-吡咯[2,3-b]吡啶-5-基)甲基)-8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4] 二氮杂
Figure PCTCN2020137640-appb-000199
-2-甲酰胺；ESI-MS(m/z)：453.3[M+H] +。 Take 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4] Diaza
Figure PCTCN2020137640-appb-000198
-2-carboxylic acid and (1H-pyrrole[2,3-b]pyridin-5-yl)methylamine as raw materials, the operation process is the same as step b in implementation 22 to obtain N-((1H-pyrrole[2,3 -b]pyridin-5-yl)methyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole [1,2-a][1,4] Diaza
Figure PCTCN2020137640-appb-000199
-2-carboxamide; ESI-MS (m/z): 453.3 [M+H] + .
实施例45Example 45
N-(2-(3-氯苯氧基)乙基)-8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000200
-2-甲酰胺(化合物45)的制备 N-(2-(3-chlorophenoxy)ethyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo [e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000200
Preparation of -2-carboxamide (Compound 45)
Figure PCTCN2020137640-appb-000201
Figure PCTCN2020137640-appb-000201
以8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000202
-2-羧酸和2-(3-氯苯氧基)乙基-1-胺为原料，操作过程同实施22中的步骤b，得N-(2-(3-氯苯氧基)乙基)-8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000203
-2-甲酰胺， 1H NMR(400MHz,DMSO-d 6)δ7.96(t,J＝6.0Hz,1H),7.77-7.59(m,1H),7.37-7.18(m,2H),7.02(t,J＝2.0Hz,1H),7.00-6.88(m,3H),6.33-6.26(m,2H),6.23(t,J＝4.4Hz,1H),6.02(d,J＝0.8Hz,1H),5.28(s,2H),5.15-4.99(m,2H),4.39(d,J＝5.2Hz,2H),4.06(t,J＝6.0Hz,2H),3.54(q,J＝5.6Hz,2H),2.21-2.07(m,3H)；ESI-MS(m/z)：477.2[M+H] +。 Take 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4] Diaza
Figure PCTCN2020137640-appb-000202
-2-carboxylic acid and 2-(3-chlorophenoxy)ethyl-1-amine as raw materials, and the operation process is the same as step b in implementation 22 to obtain N-(2-(3-chlorophenoxy)ethane Yl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1, 4] Diaza
Figure PCTCN2020137640-appb-000203
-2-Carboxamide, 1 H NMR (400MHz, DMSO-d 6 ) δ 7.96 (t, J = 6.0 Hz, 1H), 7.77-7.59 (m, 1H), 7.37-7.18 (m, 2H), 7.02 (t,J=2.0Hz,1H),7.00-6.88(m,3H),6.33-6.26(m,2H),6.23(t,J=4.4Hz,1H),6.02(d,J=0.8Hz, 1H), 5.28 (s, 2H), 5.15-4.99 (m, 2H), 4.39 (d, J = 5.2 Hz, 2H), 4.06 (t, J = 6.0 Hz, 2H), 3.54 (q, J = 5.6 Hz, 2H), 2.21-2.07 (m, 3H); ESI-MS (m/z): 477.2 [M+H] + .
实施例46Example 46
N-(4-(氨甲基)苄基)-8-((4-氟-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000204
-2-甲酰胺(化合物46)的制备 N-(4-(aminomethyl)benzyl)-8-((4-fluoro-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo(e)imidazole [1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000204
Preparation of -2-carboxamide (Compound 46)
Figure PCTCN2020137640-appb-000205
Figure PCTCN2020137640-appb-000205
操作过程同实施例41，只是将8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000206
-2-羧酸用8-((4-氟-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000207
-2-羧酸替代，得N-(4-(氨甲基)苄基)-8-((4-氟-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000208
-2-甲酰胺；ESI-MS(m/z)：446.3[M+H] +。 The procedure is the same as in Example 41, except that 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2 -a][1,4]diazepine
Figure PCTCN2020137640-appb-000206
-2-carboxylic acid 8-((4-fluoro-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][ 1,4] Diaza
Figure PCTCN2020137640-appb-000207
-2-carboxylic acid substitution, N-(4-(aminomethyl)benzyl)-8-((4-fluoro-1H-pyrazol-1-yl)methyl)-10,11-dihydro- 5H-Benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000208
-2-carboxamide; ESI-MS (m/z): 446.3 [M+H] + .
实施例47Example 47
N-(4-(氨甲基)苄基)-8-((2-氧代吡啶-1(2H)-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000209
-2-甲酰胺(化合物47)的制备 N-(4-(Aminomethyl)benzyl)-8-((2-oxopyridine-1(2H)-yl)methyl)-10,11-dihydro-5H-benzo(e)imidazole [1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000209
Preparation of -2-carboxamide (Compound 47)
Figure PCTCN2020137640-appb-000210
Figure PCTCN2020137640-appb-000210
操作过程同实施例41，只是将8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000211
-2-羧酸用8-((2-氧代吡啶-1(2H)-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000212
-2-羧酸替代，得N-(4-(氨甲基)苄基)-8-((2-氧代吡啶-1(2H)-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000213
-2-甲酰胺；ESI-MS(m/z)：455.3[M+H] +。 The procedure is the same as in Example 41, except that 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2 -a][1,4]diazepine
Figure PCTCN2020137640-appb-000211
-2-carboxylic acid 8-((2-oxopyridine-1(2H)-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][ 1,4] Diaza
Figure PCTCN2020137640-appb-000212
-2-carboxylic acid substitution, N-(4-(aminomethyl)benzyl)-8-((2-oxopyridine-1(2H)-yl)methyl)-10,11-dihydro- 5H-Benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000213
-2-carboxamide; ESI-MS (m/z): 455.3 [M+H] + .
实施例48Example 48
N-(4-(氨甲基)苄基)-8-((2-氧代噁唑烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000214
-2-甲酰胺(化合物48)的制备 N-(4-(aminomethyl)benzyl)-8-((2-oxazolidin-3-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[ 1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000214
Preparation of -2-carboxamide (Compound 48)
Figure PCTCN2020137640-appb-000215
Figure PCTCN2020137640-appb-000215
操作过程同实施例41，只是将8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000216
-2-羧酸用8-((2-氧代噁唑烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000217
-2-羧酸替代，得N-(4-(氨甲基)苄基)-8-((2-氧代噁唑烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000218
-2-甲酰胺；ESI-MS(m/z)：447.3[M+H] +。 The procedure is the same as in Example 41, except that 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2 -a][1,4]diazepine
Figure PCTCN2020137640-appb-000216
-2-carboxylic acid 8-((2-oxazolidin-3-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1 ,4] Diaza
Figure PCTCN2020137640-appb-000217
-2-carboxylic acid substitution, N-(4-(aminomethyl)benzyl)-8-((2-oxooxazolidine-3-yl)methyl)-10,11-dihydro-5H -Benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000218
-2-carboxamide; ESI-MS (m/z): 447.3 [M+H] + .
实施例49Example 49
N-(4-(氨甲基)苄基)-8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000219
-2-甲酰胺(化合物49)的制备 N-(4-(Aminomethyl)benzyl)-8-((2-oxo-3-azabicyclo[3.1.0]hexane-3-yl)methyl)-10,11-dihydro -5H-Benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000219
Preparation of -2-carboxamide (Compound 49)
Figure PCTCN2020137640-appb-000220
Figure PCTCN2020137640-appb-000220
操作过程同实施例41，只是将8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000221
-2-羧酸用8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000222
-2-羧酸替代，得N-(4-(氨甲基)苄基)-8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000223
-2-甲酰胺；ESI-MS(m/z)：457.3[M+H] +。 The procedure is the same as in Example 41, except that 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2 -a][1,4]diazepine
Figure PCTCN2020137640-appb-000221
-2-carboxylic acid 8-((2-oxo-3-azabicyclo[3.1.0]hexane-3-yl)methyl)-10,11-dihydro-5H-benzo[e] Imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000222
-2-carboxylic acid substitution to obtain N-(4-(aminomethyl)benzyl)-8-((2-oxo-3-azabicyclo[3.1.0]hexane-3-yl)methyl )-10,11-Dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000223
-2-carboxamide; ESI-MS (m/z): 457.3 [M+H] + .
实施例50Example 50
N-(4-(氨甲基)苄基)-8-(吡咯烷-1-基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000224
-2-甲酰胺(化合物50)的制备 N-(4-(Aminomethyl)benzyl)-8-(pyrrolidin-1-yl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1, 4] Diaza
Figure PCTCN2020137640-appb-000224
Preparation of -2-carboxamide (Compound 50)
Figure PCTCN2020137640-appb-000225
Figure PCTCN2020137640-appb-000225
操作过程同实施例41，只是将8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000226
-2-羧酸用8-(吡咯烷-1-基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000227
-2-羧酸替代，得N-(4-(氨甲基)苄基)-8-(吡咯烷-1-基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000228
-2-甲酰胺；ESI-MS(m/z)：417.3[M+H] +。 The procedure is the same as in Example 41, except that 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2 -a][1,4]diazepine
Figure PCTCN2020137640-appb-000226
-2-carboxylic acid 8-(pyrrolidin-1-yl)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000227
-2-carboxylic acid substitution, N-(4-(aminomethyl)benzyl)-8-(pyrrolidin-1-yl)-10,11-dihydro-5H-benzo[e]imidazole [1 ,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000228
-2-carboxamide; ESI-MS (m/z): 417.3 [M+H] + .
实施例51Example 51
N-(4-(氨甲基)苄基)-8-(苯胺基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000229
-2-甲酰胺(化合物51)的制备 N-(4-(Aminomethyl)benzyl)-8-(anilino)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4] diazonium miscellaneous
Figure PCTCN2020137640-appb-000229
Preparation of -2-carboxamide (Compound 51)
Figure PCTCN2020137640-appb-000230
Figure PCTCN2020137640-appb-000230
操作过程同实施例41，只是将8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000231
-2-羧酸用8-(苯胺基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000232
-2-羧酸替代，得N-(4-(氨甲基)苄基)-8-(苯胺基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000233
-2-甲酰胺；ESI-MS(m/z)：439.3[M+H] +。 The procedure is the same as in Example 41, except that 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2 -a][1,4]diazepine
Figure PCTCN2020137640-appb-000231
-2-carboxylic acid 8-(anilino)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000232
-2-carboxylic acid substitution, N-(4-(aminomethyl)benzyl)-8-(anilino)-10,11-dihydro-5H-benzo[e]imidazole[1,2-a ][1,4]diazepine
Figure PCTCN2020137640-appb-000233
-2-carboxamide; ESI-MS (m/z): 439.3 [M+H] + .
实施例52Example 52
N-(4-(氨甲基)苄基)-8-苯氧基-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000234
-2-甲酰胺(化合物52)的制备 N-(4-(Aminomethyl)benzyl)-8-phenoxy-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000234
Preparation of -2-carboxamide (Compound 52)
Figure PCTCN2020137640-appb-000235
Figure PCTCN2020137640-appb-000235
操作过程同实施例41，只是将8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000236
-2-羧酸用8-苯氧基-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000237
-2-羧酸替代，得N-(4-(氨甲基)苄基)-8-苯氧基-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000238
-2-甲酰胺；ESI-MS(m/z)：440.3[M+H] +。 The procedure is the same as in Example 41, except that 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]imidazole[1,2 -a][1,4]diazepine
Figure PCTCN2020137640-appb-000236
-2-carboxylic acid 8-phenoxy-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000237
-2-carboxylic acid substitution, N-(4-(aminomethyl)benzyl)-8-phenoxy-10,11-dihydro-5H-benzo[e]imidazole[1,2-a] [1,4] Diaza
Figure PCTCN2020137640-appb-000238
-2-carboxamide; ESI-MS (m/z): 440.3 [M+H] + .
实施例53Example 53
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((5-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000239
-2-甲酰胺(化合物53)的制备 N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-11-oxy Substitute -10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000239
Preparation of -2-carboxamide (Compound 53)
Figure PCTCN2020137640-appb-000240
Figure PCTCN2020137640-appb-000240
步骤a)：3-溴-1-(4-(氯甲基)-3-硝基苄基)-5-甲基-1H-吡唑的制备Step a): Preparation of 3-bromo-1-(4-(chloromethyl)-3-nitrobenzyl)-5-methyl-1H-pyrazole
将甲磺酰氯(6.21mL，54.209mmol)滴加至(4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-2-硝基苯甲醇(8.0g，24.529mmol)和三乙胺(10.2mL，73.587mmol)的二氯甲烷(100mL)溶液中，25℃反应12h，反应结束后，反应液用冰水(100mL)稀释，用二氯甲烷(100mL×3)萃取，合并有机层，饱和氯化钠水溶液(80mL×3)洗涤，无水硫酸钠干燥，过滤，滤液减压蒸干，得3-溴-1-(4-(氯甲基)-3-硝基苄基)-5-甲基-1H-吡唑，无需纯化直接用于下一步反应；ESI-MS(m/z)：344.0[M+H] +。 Methanesulfonyl chloride (6.21mL, 54.209mmol) was added dropwise to (4-((3-bromo-5-methyl-1H-pyrazol-1-yl)methyl)-2-nitrobenzyl alcohol (8.0g , 24.529mmol) and triethylamine (10.2mL, 73.587mmol) in dichloromethane (100mL) solution at 25℃ for 12h. After the reaction is over, the reaction solution is diluted with ice water (100mL) and dichloromethane (100mL) ×3) Extract, combine the organic layers, wash with saturated aqueous sodium chloride solution (80mL×3), dry with anhydrous sodium sulfate, filter, and evaporate the filtrate under reduced pressure to obtain 3-bromo-1-(4-(chloromethyl) -3-nitrobenzyl)-5-methyl-1H-pyrazole, used directly in the next reaction without purification; ESI-MS (m/z): 344.0 [M+H] + .
步骤b)：1-(4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-2-硝基苄基)-1H-吡咯-2,4-二甲酸乙酯的制备Step b): 1-(4-((3-Bromo-5-methyl-1H-pyrazol-1-yl)methyl)-2-nitrobenzyl)-1H-pyrrole-2,4-di Preparation of ethyl formate
将1H-吡咯-2,4-二甲酸乙酯(2.45g，11.608mmol)、(4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-2-硝基苯甲醇(5.0g，14.510mmol)、碳酸铯(14.2g，43.530mmol)和KI(240mg，1.451mmol)的DMF(50mL)溶液中，25℃反应5h，反应结束后，反应液用冰水(100mL)稀释，用乙酸乙酯(100mL×2)萃取，合并有机层，饱和氯化钠水溶液(80mL×3)洗涤，无水硫酸钠干燥，过滤，滤液减压蒸干，得1-(4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-2-硝基苄基)-1H-吡咯-2,4-二甲酸乙酯，无需纯化直接用于下一步反应；ESI-MS(m/z)：519.1[M+H] +。 Ethyl 1H-pyrrole-2,4-dicarboxylate (2.45g, 11.608mmol), (4-((3-bromo-5-methyl-1H-pyrazol-1-yl)methyl)-2- In a DMF (50mL) solution of nitrobenzyl alcohol (5.0g, 14.510mmol), cesium carbonate (14.2g, 43.530mmol) and KI (240mg, 1.451mmol), react at 25°C for 5 hours. After the reaction is complete, the reaction solution is heated with ice Dilute with water (100mL), extract with ethyl acetate (100mL×2), combine the organic layers, wash with saturated sodium chloride aqueous solution (80mL×3), dry with anhydrous sodium sulfate, filter, and evaporate the filtrate under reduced pressure to obtain 1- (4-((3-Bromo-5-methyl-1H-pyrazol-1-yl)methyl)-2-nitrobenzyl)-1H-pyrrole-2,4-dicarboxylate, without purification It was directly used in the next reaction; ESI-MS (m/z): 519.1 [M+H] + .
步骤c-e)：8-((5-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000241
-2-甲酸乙酯的制备 Step ce): 8-((5-methyl-1H-pyrazol-1-yl)methyl)-11-oxo-10,11-dihydro-5H-benzo[e]pyrrole [1,2 -a][1,4]diazepine
Figure PCTCN2020137640-appb-000241
Preparation of -2-ethyl formate
以1-(4-((3-溴-5-甲基-1H-吡唑-1-基)甲基)-2-硝基苄基)-1H-吡咯-2,4-二甲酸乙酯为原料，其他操作过程同实施例1中的步骤k-m，得8-((5-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000242
-2-甲酸乙酯；ESI-MS(m/z)：365.2[M+H] +。 Ethyl 1-(4-((3-bromo-5-methyl-1H-pyrazol-1-yl)methyl)-2-nitrobenzyl)-1H-pyrrole-2,4-dicarboxylate As the raw material, the other operation process is the same as the step km in Example 1, to obtain 8-((5-methyl-1H-pyrazol-1-yl)methyl)-11-oxo-10,11-dihydro- 5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000242
Ethyl-2-carboxylate; ESI-MS (m/z): 365.2 [M+H] + .
步骤f)：8-((5-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000243
-2-羧酸的制备 Step f): 8-((5-methyl-1H-pyrazol-1-yl)methyl)-11-oxo-10,11-dihydro-5H-benzo[e]pyrrole[1,2 -a][1,4]diazepine
Figure PCTCN2020137640-appb-000243
Preparation of -2-carboxylic acid
将LiOH.H 2O(25.3mg，0.602mmol)的水溶液(1mL)加入8-((5-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000244
-2-甲酸乙酯(110mg，0.301mmol)的甲醇(1mL)/四氢呋喃(1mL)溶液中，25℃反应12h，反应结束后，反应液冷冻干燥，所得固体以二氯甲烷/甲醇混合液(20mL，1:1)溶解，过滤除去不溶物，滤液减压浓缩，得8-((5-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000245
-2-羧酸；ESI-MS(m/z)：337.2[M+H] +。 Add an aqueous solution (1 mL) of LiOH.H 2 O (25.3 mg, 0.602 mmol) to 8-((5-methyl-1H-pyrazol-1-yl)methyl)-11-oxo-10,11- Dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000244
-2-ethyl formate (110mg, 0.301mmol) in methanol (1mL)/tetrahydrofuran (1mL) solution, reacted at 25°C for 12h. After the reaction, the reaction solution was freeze-dried. 20mL, 1:1) dissolved, filtered to remove insolubles, and the filtrate was concentrated under reduced pressure to obtain 8-((5-methyl-1H-pyrazol-1-yl)methyl)-11-oxo-10,11- Dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000245
-2-carboxylic acid; ESI-MS (m/z): 337.2 [M+H] + .
步骤g)：N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((5-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000246
-2-甲酰胺的制备 Step g): N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl) -11-oxo-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000246
Preparation of -2-carboxamide
以8-((5-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000247
-2-羧酸和5-(氨甲基)-4,6-二甲基吡啶-2-胺为原料，其他操作过程同实施例1中的步骤p，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((5-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000248
-2-甲酰胺，收率39.1％， 1H NMR(400MHz,DMSO-d 6)δ10.17(s,1H),8.11(t,J＝4.8Hz,1H),7.48-7.59(m,3H),7.32-7.38(m,2H),7.21(d,J＝2.0Hz,1H),6.94(s,1H),6.84-6.89(m,1H),6.62(s,1H),6.07(s,1H),5.25(s,2H),5.17(s,2H),4.26(d,J＝4.8Hz,2H),2.49(br s,3H),2.36(s,3H),2.20(s,3H)；ESI-MS(m/z)：470.3[M+H] +。 As 8-((5-methyl-1H-pyrazol-1-yl)methyl)-11-oxo-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a] [1,4] Diaza
Figure PCTCN2020137640-appb-000247
-2-carboxylic acid and 5-(aminomethyl)-4,6-dimethylpyridin-2-amine as raw materials, and other operating procedures are the same as those in step p in Example 1, to obtain N-((6-amino- 2,4-dimethylpyridin-3-yl)methyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-11-oxo-10,11-dihydro -5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000248
-2-carboxamide, yield 39.1%, 1 H NMR (400MHz, DMSO-d 6 )δ10.17(s,1H), 8.11(t,J=4.8Hz,1H), 7.48-7.59(m,3H ), 7.32-7.38 (m, 2H), 7.21 (d, J = 2.0Hz, 1H), 6.94 (s, 1H), 6.84-6.89 (m, 1H), 6.62 (s, 1H), 6.07 (s, 1H), 5.25 (s, 2H), 5.17 (s, 2H), 4.26 (d, J = 4.8 Hz, 2H), 2.49 (br s, 3H), 2.36 (s, 3H), 2.20 (s, 3H) ; ESI-MS (m/z): 470.3 [M+H] + .
实施例54Example 54
8-((1H-吡唑-1-基)甲基)-N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000249
-2-甲酰胺(化合物54)的制备 8-((1H-pyrazol-1-yl)methyl)-N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-10,11 -Dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000249
Preparation of -2-carboxamide (Compound 54)
Figure PCTCN2020137640-appb-000250
Figure PCTCN2020137640-appb-000250
步骤a-b)：1-(4-((1H-吡唑-1-基)甲基)-2-硝基苄基)-1H-吡咯-2,4-二甲酸乙酯的制备Step a-b): Preparation of 1-(4-((1H-pyrazol-1-yl)methyl)-2-nitrobenzyl)-1H-pyrrole-2,4-dicarboxylate ethyl ester
以(4-((1H-吡唑-1-基)甲基)-2-硝基)苯甲醇为起始原料，其他操作过程同实施例53中的步骤a-b，得1-(4-((1H-吡唑-1-基)甲基)-2-硝基苄基)-1H-吡咯-2,4-二甲酸乙酯；ESI-MS(m/z)：427.2[M+H] +。 Using (4-((1H-pyrazol-1-yl)methyl)-2-nitro)benzyl alcohol as the starting material, the other operating procedures were the same as the steps ab in Example 53, to obtain 1-(4-( (1H-Pyrazol-1-yl)methyl)-2-nitrobenzyl)-1H-pyrrole-2,4-dicarboxylate; ESI-MS(m/z): 427.2[M+H] + .
步骤c-d)：8-((1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000251
-2-甲酸乙酯的制备 Step cd): 8-((1H-pyrazol-1-yl)methyl)-11-oxo-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1 ,4] Diaza
Figure PCTCN2020137640-appb-000251
Preparation of -2-ethyl formate
以1-(4-((1H-吡唑-1-基)甲基)-2-硝基苄基)-1H-吡咯-2,4-二甲酸乙酯为原料，其他操作过程同实施例2中的步骤e-f，得8-((1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000252
-2-甲酸乙酯；ESI-MS(m/z)：351.2[M+H] +。 Take 1-(4-((1H-pyrazol-1-yl)methyl)-2-nitrobenzyl)-1H-pyrrole-2,4-dicarboxylate ethyl ester as raw material, and other operation procedures are the same as in the embodiment Step ef in 2 gives 8-((1H-pyrazol-1-yl)methyl)-11-oxo-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a ][1,4]diazepine
Figure PCTCN2020137640-appb-000252
Ethyl-2-carboxylate; ESI-MS (m/z): 351.2 [M+H] + .
步骤e-f)：8-((1H-吡唑-1-基)甲基)-N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000253
-2-甲酰胺的制备 Step ef): 8-((1H-pyrazol-1-yl)methyl)-N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo -10,11-Dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000253
Preparation of -2-carboxamide
以8-((1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000254
-2-甲酸乙酯为原料，其他操作过程同实施例53中的步骤f-g，得8-((1H-吡唑-1-基)甲基)-N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000255
-2-甲酰胺；ESI-MS(m/z)：456.3[M+H] +。 As 8-((1H-pyrazol-1-yl)methyl)-11-oxo-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4] Diaza
Figure PCTCN2020137640-appb-000254
Ethyl -2-formate was used as the raw material, and the other operating procedures were the same as those in step fg in Example 53, to obtain 8-((1H-pyrazol-1-yl)methyl)-N-((6-amino-2,4 -Dimethylpyridin-3-yl)methyl)-11-oxo-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000255
-2-carboxamide; ESI-MS (m/z): 456.3 [M+H] + .
实施例55Example 55
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((4-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000256
-2-甲酰胺(化合物55)的制备 N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((4-methyl-1H-pyrazol-1-yl)methyl)-11-oxy Substitute -10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000256
Preparation of -2-carboxamide (Compound 55)
Figure PCTCN2020137640-appb-000257
Figure PCTCN2020137640-appb-000257
以(4-((4-甲基-1H-吡唑-1-基)甲基)-2-硝基)苯甲醇为原料，其他操作过程同实施例54，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((4-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000258
-2-甲酰胺；ESI-MS(m/z)：470.3[M+H] +。 Using (4-((4-methyl-1H-pyrazol-1-yl)methyl)-2-nitro)benzyl alcohol as a raw material, the other procedures were the same as those in Example 54, to obtain N-((6-amino -2,4-dimethylpyridin-3-yl)methyl)-8-((4-methyl-1H-pyrazol-1-yl)methyl)-11-oxo-10,11-di Hydrogen-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000258
-2-carboxamide; ESI-MS (m/z): 470.3 [M+H] + .
实施例56Example 56
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((4-氟-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯 并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000259
-2-甲酰胺(化合物56)的制备 N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((4-fluoro-1H-pyrazol-1-yl)methyl)-11-oxo -10,11-Dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000259
Preparation of -2-carboxamide (Compound 56)
Figure PCTCN2020137640-appb-000260
Figure PCTCN2020137640-appb-000260
以(4-((4-氟-1H-吡唑-1-基)甲基)-2-硝基)苯甲醇为原料，其他操作过程同实施例54，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((4-氟-1H-吡唑-1-基)甲基)-11-氧代-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000261
-2-甲酰胺；ESI-MS(m/z)：474.3[M+H] +。 Using (4-((4-fluoro-1H-pyrazol-1-yl)methyl)-2-nitro)benzyl alcohol as the raw material, and the other operating procedures were the same as those in Example 54, N-((6-amino- 2,4-dimethylpyridin-3-yl)methyl)-8-((4-fluoro-1H-pyrazol-1-yl)methyl)-11-oxo-10,11-dihydro- 5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000261
-2-carboxamide; ESI-MS (m/z): 474.3 [M+H] + .
实施例57Example 57
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-((2-氧代吡啶-1(2H)-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000262
-2-甲酰胺(化合物57)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8-((2-oxopyridin-1(2H)-yl)methyl) -10,11-Dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000262
Preparation of -2-carboxamide (Compound 57)
Figure PCTCN2020137640-appb-000263
Figure PCTCN2020137640-appb-000263
以1-(4-(羟甲基)-3-硝基苄基)吡啶-2(1H)-酮为原料，其他操作过程同实施例54，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-((2-氧代吡啶-1(2H)-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000264
-2-甲酰胺；ESI-MS(m/z)：483.3[M+H] +。 Using 1-(4-(hydroxymethyl)-3-nitrobenzyl)pyridine-2(1H)-one as the raw material, the other procedures are the same as in Example 54, to obtain N-((6-amino-2,4 -Dimethylpyridin-3-yl)methyl)-11-oxo-8-((2-oxopyridin-1(2H)-yl)methyl)-10,11-dihydro-5H-benzene And [e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000264
-2-carboxamide; ESI-MS (m/z): 483.3 [M+H] + .
实施例58Example 58
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-((2-氧代噁唑烷-3-基)甲基)-10,11--二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000265
-2-甲酰胺(化合物58)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8-((2-oxooxazolidin-3-yl)methyl)- 10,11--Dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000265
Preparation of -2-carboxamide (Compound 58)
Figure PCTCN2020137640-appb-000266
Figure PCTCN2020137640-appb-000266
以3-(4-(羟甲基)-3-硝基苄基)噁唑烷-2-酮为原料，其他操作过程同实施例54，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-((2-氧代噁唑烷-3-基)甲基)-10,11--二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000267
-2-甲酰胺；ESI-MS(m/z)：475.3[M+H] +。 Using 3-(4-(hydroxymethyl)-3-nitrobenzyl)oxazolidin-2-one as the raw material, the other operating procedures were the same as those in Example 54, to obtain N-((6-amino-2,4- Dimethylpyridin-3-yl)methyl)-11-oxo-8-((2-oxooxazolidin-3-yl)methyl)-10,11--dihydro-5H-benzo [e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000267
-2-carboxamide; ESI-MS (m/z): 475.3 [M+H] + .
实施例59Example 59
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000268
-2-甲酰胺(化合物59)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8-((2-oxo-3-azabicyclo[3.1.0]hexane Alkyl-3-yl)methyl)-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000268
Preparation of -2-carboxamide (Compound 59)
Figure PCTCN2020137640-appb-000269
Figure PCTCN2020137640-appb-000269
以3-(4-(羟甲基)-3-硝基苄基)-3-氮杂双环[3.1.0]己烷-2-酮为原料，其他操作过程同实施例54，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基) 甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000270
-2-甲酰胺；ESI-MS(m/z)：485.3[M+H] +。 Using 3-(4-(hydroxymethyl)-3-nitrobenzyl)-3-azabicyclo[3.1.0]hexane-2-one as the raw material, other procedures were the same as in Example 54, to obtain N- ((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8-((2-oxo-3-azabicyclo[3.1.0]hexane- 3-yl)methyl)-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000270
-2-carboxamide; ESI-MS (m/z): 485.3 [M+H] + .
实施例60Example 60
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-(吡咯烷-1-基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000271
-2-甲酰胺(化合物60)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8-(pyrrolidin-1-yl)-10,11-dihydro-5H- Benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000271
Preparation of -2-carboxamide (Compound 60)
Figure PCTCN2020137640-appb-000272
Figure PCTCN2020137640-appb-000272
以2-硝基-4-(吡咯烷-1-基)苯甲醇为原料，其他操作过程同实施例54，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-(吡咯烷-1-基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000273
-2-甲酰胺；ESI-MS(m/z)：445.3[M+H] +。 Using 2-nitro-4-(pyrrolidin-1-yl)benzyl alcohol as the raw material, the other operating procedures were the same as those in Example 54, to obtain N-((6-amino-2,4-dimethylpyridin-3-yl) )Methyl)-11-oxo-8-(pyrrolidin-1-yl)-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4] diazonium miscellaneous
Figure PCTCN2020137640-appb-000273
-2-carboxamide; ESI-MS (m/z): 445.3 [M+H] + .
实施例61Example 61
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-(苯胺基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000274
-2-甲酰胺(化合物61)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8-(anilino)-10,11-dihydro-5H-benzo(e ]Pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000274
Preparation of -2-carboxamide (Compound 61)
Figure PCTCN2020137640-appb-000275
Figure PCTCN2020137640-appb-000275
以2-硝基-4-(苯胺基)苯甲醇为原料，其他操作过程同实施例54，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-(苯胺基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000276
-2-甲酰胺；ESI-MS(m/z)：467.3[M+H] +。 Using 2-nitro-4-(anilino)benzyl alcohol as the raw material, the other procedures are the same as in Example 54, to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl) -11-oxo-8-(anilino)-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000276
-2-carboxamide; ESI-MS (m/z): 467.3 [M+H] + .
实施例62Example 62
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-苯氧基-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000277
-2-甲酰胺(化合物62)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-11-oxo-8-phenoxy-10,11-dihydro-5H-benzo[e] Pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000277
Preparation of -2-carboxamide (Compound 62)
Figure PCTCN2020137640-appb-000278
Figure PCTCN2020137640-appb-000278
以2-硝基-4-(苯氧基)苯甲醇为原料，其他操作过程同实施例54，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-11-氧代-8-苯氧基-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000279
-2-甲酰胺；ESI-MS(m/z)：468.3[M+H] +。 Using 2-nitro-4-(phenoxy)benzyl alcohol as the raw material, the other operating procedures were the same as those in Example 54, to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl )-11-oxo-8-phenoxy-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000279
-2-carboxamide; ESI-MS (m/z): 468.3 [M+H] + .
实施例63Example 63
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000280
-2-甲酰胺(化合物63)的制备 N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11 -Dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000280
Preparation of -2-carboxamide (Compound 63)
Figure PCTCN2020137640-appb-000281
Figure PCTCN2020137640-appb-000281
步骤a)：8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000282
-2-甲酸乙酯的制备 Step a): 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1 ,4] Diaza
Figure PCTCN2020137640-appb-000282
Preparation of -2-ethyl formate
以8-((5-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11--二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000283
-2-甲酸乙酯为原料，其他操作过程同实施例1中的步骤n，得8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000284
-2-甲酸乙酯；ESI-MS(m/z)：351.2[M+H] +。 Take 8-((5-methyl-1H-pyrazol-1-yl)methyl)-11-oxo-10,11--dihydro-5H-benzo[e]pyrrole[1,2-a ][1,4]diazepine
Figure PCTCN2020137640-appb-000283
Ethyl -2-formate was used as the raw material, and the other procedures were the same as the step n in Example 1, to obtain 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro -5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000284
Ethyl-2-carboxylate; ESI-MS (m/z): 351.2 [M+H] + .
步骤b-c)：N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000285
-2-甲酰胺的制备 Step bc): N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((5-methyl-1H-pyrazol-1-yl)methyl) -10,11-Dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000285
Preparation of -2-carboxamide
以8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000286
-2-甲酸乙酯为原料，其他操作同实施例53中的步骤f-g，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((5-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000287
-2-甲酰胺， 1H NMR(400MHz,DMSO-d 6)δ7.88-7.81(m,1H),7.62-7.41(m,2H),7.32-7.28(m,2H),6.93-6.87(m,1H),6.63-6.60(m,1H),6.31-6.28(m,1H),6.20-6.15(m,2H),6.04-6.00(m,1H),5.21-5.16(m,2H),5.08-5.03(m,2H),4.32-4.26(m,2H),4.25-4.20(m,2H),2.55-2.52(m,3H),2.41-2.35(m,3H),2.19-2.11(m,3H)；ESI-MS(m/z)：456.2[M+H] +。 Take 8-((5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4] Diaza
Figure PCTCN2020137640-appb-000286
Ethyl-2-carboxylate as the raw material, and other operations were the same as those in step fg in Example 53, to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-(( 5-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000287
-2-Carboxamide, 1 H NMR (400MHz, DMSO-d 6 ) δ7.88-7.81(m,1H), 7.62-7.41(m,2H), 7.32-7.28(m,2H), 6.93-6.87( m, 1H), 6.63-6.60 (m, 1H), 6.31-6.28 (m, 1H), 6.20-6.15 (m, 2H), 6.04-6.00 (m, 1H), 5.21-5.16 (m, 2H), 5.08-5.03 (m, 2H), 4.32-4.26 (m, 2H), 4.25-4.20 (m, 2H), 2.55-2.52 (m, 3H), 2.41-2.35 (m, 3H), 2.19-2.11 (m , 3H); ESI-MS (m/z): 456.2 [M+H] + .
实施例64Example 64
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((4-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000288
-2-甲酰胺(化合物64)的制备 N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((4-methyl-1H-pyrazol-1-yl)methyl)-10,11 -Dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000288
Preparation of -2-carboxamide (Compound 64)
Figure PCTCN2020137640-appb-000289
Figure PCTCN2020137640-appb-000289
以8-((4-甲基-1H-吡唑-1-基)甲基)-11-氧代-10,11--二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000290
-2-甲酸乙酯为原料，其他操作过程同实施例63，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((4-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000291
-2-甲酰胺；ESI-MS(m/z)：456.3[M+H] +。 Take 8-((4-methyl-1H-pyrazol-1-yl)methyl)-11-oxo-10,11--dihydro-5H-benzo[e]pyrrole[1,2-a ][1,4]diazepine
Figure PCTCN2020137640-appb-000290
Ethyl -2-formate was used as the raw material, and the other procedures were the same as in Example 63 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((4-methyl -1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000291
-2-carboxamide; ESI-MS (m/z): 456.3 [M+H] + .
实施例65Example 65
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((2-氧代吡啶-1(2H)-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000292
-2-甲酰胺(化合物65)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-8-((2-oxopyridin-1(2H)-yl)methyl)-10,11- Dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000292
Preparation of -2-carboxamide (Compound 65)
Figure PCTCN2020137640-appb-000293
Figure PCTCN2020137640-appb-000293
以11-氧代-8-((2-氧代吡啶-1(2H)-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000294
-2-甲酸乙酯为原料，其他操作过程同实施例63，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((2-氧代吡啶-1(2H)-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000295
-2-甲酰胺；ESI-MS(m/z)：469.3[M+H] +。 Take 11-oxo-8-((2-oxopyridine-1(2H)-yl)methyl)-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][ 1,4] Diaza
Figure PCTCN2020137640-appb-000294
Ethyl-2-carboxylate was used as the raw material, and the other procedures were the same as in Example 63 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((2-oxo (Pyridine-1(2H)-yl)methyl)-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000295
-2-carboxamide; ESI-MS (m/z): 469.3 [M+H] + .
实施例66Example 66
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((2-氧代噁唑烷-3-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000296
-2-甲酰胺(化合物66)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-8-((2-oxazolidin-3-yl)methyl)-10,11-bis Hydrogen-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000296
Preparation of -2-carboxamide (Compound 66)
Figure PCTCN2020137640-appb-000297
Figure PCTCN2020137640-appb-000297
以11-氧代-8-((2-氧代噁唑烷-3-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000298
-2-甲酸乙酯为原料，其他操作过程同实施例63，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((2-氧代噁唑烷-3-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000299
-2-甲酰胺；ESI-MS(m/z)：461.3[M+H] +。 Take 11-oxo-8-((2-oxooxazolidin-3-yl)methyl)-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1 ,4] Diaza
Figure PCTCN2020137640-appb-000298
Ethyl-2-carboxylate was used as the raw material, and the other procedures were the same as in Example 63 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((2-oxo Oxazolidin-3-yl)methyl)-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000299
-2-carboxamide; ESI-MS (m/z): 461.3 [M+H] + .
实施例67Example 67
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000300
-2-甲酰胺(化合物67)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-8-((2-oxo-3-azabicyclo[3.1.0]hexane-3-yl )Methyl)-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000300
Preparation of -2-carboxamide (Compound 67)
Figure PCTCN2020137640-appb-000301
Figure PCTCN2020137640-appb-000301
以11-氧代-8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000302
-2-甲酸乙酯为原料，其他操作过程同实施例63，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((2-氧代-3-氮杂双环[3.1.0]己烷-3-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000303
-2-甲酰胺；ESI-MS(m/z)：471.3[M+H] +。 Take 11-oxo-8-((2-oxo-3-azabicyclo[3.1.0]hexane-3-yl)methyl)-10,11-dihydro-5H-benzo[e] Pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000302
Ethyl-2-carboxylate was used as the raw material, and the other procedures were the same as in Example 63 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((2-oxo -3-azabicyclo[3.1.0]hexane-3-yl)methyl)-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4] Diaza
Figure PCTCN2020137640-appb-000303
-2-carboxamide; ESI-MS (m/z): 471.3 [M+H] + .
实施例68Example 68
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-(吡咯烷-1-基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000304
-2-甲酰胺(化合物68)的制备 N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-(pyrrolidin-1-yl)-10,11-dihydro-5H-benzo[e] Pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000304
Preparation of -2-carboxamide (Compound 68)
Figure PCTCN2020137640-appb-000305
Figure PCTCN2020137640-appb-000305
以11-氧代-8-(吡咯烷-1-基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000306
-2-甲酸乙酯为原料，其他操作过程同实施例63，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-(吡咯烷-1-基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000307
-2-甲酰胺；ESI-MS(m/z)：431.3[M+H] +。 As 11-oxo-8-(pyrrolidin-1-yl)-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000306
Ethyl-2-carboxylate was used as the raw material, and the other procedures were the same as those in Example 63 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-(pyrrolidine-1) -Yl)-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000307
-2-carboxamide; ESI-MS (m/z): 431.3 [M+H] + .
实施例69Example 69
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-(苯胺基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000308
-2-甲酰胺(化合物69)的制备 N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-(anilino)-10,11-dihydro-5H-benzo[e]pyrrole[1, 2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000308
Preparation of -2-carboxamide (Compound 69)
Figure PCTCN2020137640-appb-000309
Figure PCTCN2020137640-appb-000309
以11-氧代-8-(苯胺基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000310
-2-甲酸乙酯为原料，其他操作过程同实施例63，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-(苯胺基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000311
-2-甲酰胺；ESI-MS(m/z)：453.3[M+H] +。 Take 11-oxo-8-(anilino)-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000310
Ethyl -2-formate was used as the raw material, and the other procedures were the same as in Example 63 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-(anilino)- 10,11-Dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000311
-2-carboxamide; ESI-MS (m/z): 453.3 [M+H] + .
实施例70Example 70
N-((6-氨基-2,4-二甲基吡啶-3-基)基)-8-苯氧基-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000312
-2-甲酰胺(化合物70)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)yl)-8-phenoxy-10,11-dihydro-5H-benzo[e]pyrrole[1,2- a][1,4]diazepine
Figure PCTCN2020137640-appb-000312
Preparation of -2-carboxamide (Compound 70)
Figure PCTCN2020137640-appb-000313
Figure PCTCN2020137640-appb-000313
以11-氧代-8-苯氧基-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000314
-2-甲酸乙酯为原料，其他操作过程同实施例63，得N-((6-氨基-2,4-二甲基吡啶-3-基)基)-8-苯氧基-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000315
-2-甲酰胺；ESI-MS(m/z)：454.3[M+H] +。 Taking 11-oxo-8-phenoxy-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000314
Ethyl-2-carboxylate was used as the raw material, and the other procedures were the same as in Example 63 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)yl)-8-phenoxy-10, 11-Dihydro-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000315
-2-carboxamide; ESI-MS (m/z): 454.3 [M+H] + .
实施例71Example 71
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((4-甲基-1H-吡唑-1-基)甲基)-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000316
-2-甲酰胺(化合物71)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-8-((4-methyl-1H-pyrazol-1-yl)methyl)-5H-benzene And [e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000316
Preparation of -2-carboxamide (Compound 71)
Figure PCTCN2020137640-appb-000317
Figure PCTCN2020137640-appb-000317
以8-((4-甲基-1H-吡唑-1-基)甲基)-10,11-二氢-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000318
-2-羧酸为起始原料，其他操作过程同实施例30，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-((4-甲基-1H-吡唑-1-基)甲基)-5H-苯并[e]吡咯[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000319
-2-甲酰胺；ESI-MS(m/z)：454.3[M+H] +。 Take 8-((4-methyl-1H-pyrazol-1-yl)methyl)-10,11-dihydro-5H-benzo[e]pyrrole[1,2-a][1,4] Diaza
Figure PCTCN2020137640-appb-000318
-2-carboxylic acid was the starting material, and the other procedures were the same as in Example 30 to obtain N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-8-((4- Methyl-1H-pyrazol-1-yl)methyl)-5H-benzo[e]pyrrole[1,2-a][1,4]diazepine
Figure PCTCN2020137640-appb-000319
-2-carboxamide; ESI-MS (m/z): 454.3 [M+H] + .
实施例72Example 72
N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-苯基-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000320
-2-甲酰胺(化合物72)的制备 N-((6-Amino-2,4-dimethylpyridin-3-yl)methyl)-8-phenyl-10,11-dihydro-5H-benzo[e]imidazole[1,2- a][1,4]diazepine
Figure PCTCN2020137640-appb-000320
Preparation of -2-carboxamide (Compound 72)
Figure PCTCN2020137640-appb-000321
Figure PCTCN2020137640-appb-000321
以(3-硝基-[1,1'-联苯基]-4-基)甲醇为起始原料，其他操作过程同实施例2中的步骤c-i，得N-((6-氨基-2,4-二甲基吡啶-3-基)甲基)-8-苯基-10,11-二氢-5H-苯并[e]咪唑[1,2-a][1,4]二氮杂
Figure PCTCN2020137640-appb-000322
-2-甲酰胺；ESI-MS(m/z)：439.3[M+H] +。 Using (3-nitro-[1,1'-biphenyl]-4-yl)methanol as the starting material, the other operating procedures are the same as the step ci in Example 2, to obtain N-((6-amino-2 ,4-dimethylpyridin-3-yl)methyl)-8-phenyl-10,11-dihydro-5H-benzo[e]imidazole[1,2-a][1,4]diazepine miscellaneous
Figure PCTCN2020137640-appb-000322
-2-carboxamide; ESI-MS (m/z): 439.3 [M+H] + .
实施例73Example 73
N-((R)-1-(((S)-1-((4-(氨甲基)苄基)氨基)-1-氧代-3-苯丙基-2-基)氨基)-3-(4-乙氧基苯基)-1-氧丙基-2-基)苯甲酰胺(化合物73)的制备N-((R)-1-(((S)-1-((4-(aminomethyl)benzyl)amino)-1-oxo-3-phenylpropyl-2-yl)amino)- Preparation of 3-(4-ethoxyphenyl)-1-oxopropyl-2-yl)benzamide (Compound 73)
Figure PCTCN2020137640-appb-000323
Figure PCTCN2020137640-appb-000323
步骤a)：(R)-2-氨基-3-(4-乙氧基苯基)丙酸盐酸盐的制备Step a): Preparation of (R)-2-amino-3-(4-ethoxyphenyl)propionate hydrochloride
将(R)-2-((叔丁氧羰基)氨基)-3-(4-乙氧基苯基)丙酸(2.0g，6.465mmol)置于反应瓶中，加入自制的盐酸二氧六环溶液(5.3M，50mL)，室温反应3小时，过滤收集产生的白色固体，滤饼用二氧六环(20mL)，石油醚(50mL)，乙醚(10mL)洗涤，真空干燥得到白色固体产品，收率98.0％，ESI-MS(m/z)：210.1[M+H] +。 Put (R)-2-((tert-butoxycarbonyl)amino)-3-(4-ethoxyphenyl)propionic acid (2.0g, 6.465mmol) in the reaction flask, add homemade dioxane hydrochloride The ring solution (5.3M, 50mL) was reacted at room temperature for 3 hours. The white solid produced was collected by filtration. The filter cake was washed with dioxane (20mL), petroleum ether (50mL), diethyl ether (10mL), and vacuum dried to obtain a white solid product. , The yield is 98.0%, ESI-MS (m/z): 210.1 [M+H] + .
步骤b)：((苄氧)羰基)-D-苯丙氨酸的制备Step b): Preparation of ((benzyloxy)carbonyl)-D-phenylalanine
将(R)-2-氨基-3-(4-乙氧基苯基)丙酸盐酸盐(500mg，2.035mmol)和氢氧化钠(179mg， 4.478mmol)加入反应瓶中，加水(15mL)搅拌溶解，冷却至0℃，缓慢滴加氯甲酸苄酯(382mg，2.239mmol)的二氧六环溶液(15mL)，滴加完毕，室温搅拌反应过夜，反应结束后，蒸除溶剂，加水稀释(30mL)，再加入乙醚(30mL)萃取，弃去有机相，水层用1N盐酸水溶液调节pH＝3-4，以乙酸乙酯(50mL×2)萃取，合并有机层，饱和氯化钠水溶液(30mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，得((苄氧)羰基)-D-苯丙氨酸，收率99.0％，ESI-MS(m/z)：300.1[M+H] +。 Add (R)-2-amino-3-(4-ethoxyphenyl)propionate hydrochloride (500mg, 2.035mmol) and sodium hydroxide (179mg, 4.478mmol) into the reaction flask, add water (15mL) Stir to dissolve, cool to 0°C, slowly add dioxane solution (15mL) of benzyl chloroformate (382mg, 2.239mmol) dropwise, after the dropwise addition is complete, stir at room temperature overnight, after the reaction is over, distill off the solvent and add water to dilute (30mL), then add ether (30mL) for extraction, discard the organic phase, adjust the pH to 3-4 with 1N aqueous hydrochloric acid solution, extract with ethyl acetate (50mL×2), combine the organic layers, and saturated sodium chloride aqueous solution (30mL) washed, dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain ((benzyloxy)carbonyl)-D-phenylalanine with a yield of 99.0%, ESI-MS(m/z): 300.1[ M+H] + .
步骤c)：(S)-(1-((4-(((叔丁氧羰基)氨基)甲基)苄基)氨基)-1-氧-3-苯丙基-2-基)氨基甲酸苄酯的制备Step c): (S)-(1-((4-(((tert-butoxycarbonyl)amino)methyl)benzyl)amino)-1-oxo-3-phenylpropyl-2-yl)carbamic acid Preparation of benzyl ester
将((苄氧基)羰基)-L-苯丙氨酸(1.39g，4.655mmol)、(4-(氨甲基)苄基)氨基甲酸叔丁酯(1.0g，4.232mmol)、HATU(3.22g，8.464mmol)、三乙胺(1.77mL，12.696mmol)和干燥二氯甲烷(30mL)加入反应瓶中，氮气保护下室温搅拌反应4h，反应结束后，向反应液中加入二氯甲烷(50mL)和饱和氯化铵水溶液(100mL)萃取，水层再以二氯甲烷(50mL)萃取，合并有机层，依次用饱和碳酸钾水溶液(100mL)和饱和氯化钠水溶液(100mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，所得粗品分散到石油醚和乙酸乙酯混合液(v/v＝8:1，25mL)中，搅拌25min，抽滤，滤饼以相应的混合液洗涤，40℃减压真空干燥，得类白色固体，收率97.0％；ESI-MS(m/z)：518.3[M+H] +。 The ((benzyloxy)carbonyl)-L-phenylalanine (1.39g, 4.655mmol), (4-(aminomethyl)benzyl) tert-butyl carbamate (1.0g, 4.232mmol), HATU( 3.22g, 8.464mmol), triethylamine (1.77mL, 12.696mmol) and dry dichloromethane (30mL) were added to the reaction flask, and the reaction was stirred at room temperature under nitrogen protection for 4h. After the reaction, dichloromethane was added to the reaction solution. (50mL) and saturated aqueous ammonium chloride solution (100mL). The aqueous layer was extracted with dichloromethane (50mL). The organic layers were combined and washed with saturated potassium carbonate aqueous solution (100mL) and saturated sodium chloride aqueous solution (100mL) successively. Dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and disperse the obtained crude product into a mixture of petroleum ether and ethyl acetate (v/v=8:1, 25mL), stir for 25min, filter with suction, and mix the filter cake accordingly The liquid was washed and dried under reduced pressure at 40°C to obtain an off-white solid with a yield of 97.0%; ESI-MS (m/z): 518.3 [M+H] + .
步骤d)：(S)-(4-((2-氨基-3-苯丙氨基)甲基)苄基)氨基甲酸叔丁酯的制备Step d): Preparation of tert-butyl (S)-(4-((2-amino-3-phenylpropylamino)methyl)benzyl)carbamate
将(S)-(1-((4-(((叔丁氧羰基)氨基)甲基)苄基)氨基)-1-氧-3-苯丙基-2-基)氨基甲酸苄酯(2.0g，3.864mmol)和甲醇(150mL)加入反应瓶中，搅拌混匀，加入Pd/C(400mg)，通氢气室温搅拌反应24h，反应结束后，过滤，滤饼用甲醇(100mL)洗涤，合并滤液，减压浓缩，得类白色固体，收率92.0％，ESI-MS(m/z)：384.3[M+H] +。 (S)-(1-((4-(((tert-butoxycarbonyl)amino)methyl)benzyl)amino)-1-oxo-3-phenylpropyl-2-yl)benzyl carbamate ( 2.0g, 3.864mmol) and methanol (150mL) were added to the reaction flask, stirred and mixed, Pd/C (400mg) was added, hydrogen gas was stirred at room temperature and reacted for 24h. After the reaction was completed, filtered, and the filter cake was washed with methanol (100mL). The filtrates were combined and concentrated under reduced pressure to obtain an off-white solid with a yield of 92.0%. ESI-MS (m/z): 384.3 [M+H] + .
步骤e)：(4-((5R,8S)-8-苄基-5-(4-乙氧基苄基)-3,6,9-三氧代-1-苯基-2-氧-4,7,10-三氮十一烷-11-基)苄基)氨基甲酸叔丁酯的制备Step e): (4-((5R,8S)-8-benzyl-5-(4-ethoxybenzyl)-3,6,9-trioxo-1-phenyl-2-oxo- Preparation of 4,7,10-triazaundecane-11-yl)benzyl) tert-butyl carbamate
将((苄氧)羰基)-D-苯丙氨酸(257mg，0.860mmol)、(S)-(4-((2-氨基-3-苯丙氨基)甲基)苄基)氨基甲酸叔丁酯(300mg，0.782mmol)、HATU(595mg，1.564mmol)、三乙胺(237mg，2.346mmol)和二氯甲烷加入反应瓶中(10mL)中，室温反应过夜，反应结束后，向反应液中加入二氯甲烷(100mL)稀释，依次用1N盐酸水溶液(100mL)、饱和氯化钠水溶液(100mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，所得粗品以石油醚/乙酸乙酯混合溶剂(9:1，v/v，50mL)打浆，过滤，减压真空干燥得类白色的固体，收率90.5％；ESI-MS(m/z)：709.4[M+H] +。 ((Benzyloxy)carbonyl)-D-phenylalanine (257mg, 0.860mmol), (S)-(4-((2-amino-3-phenylpropylamino)methyl)benzyl)carbamic acid tert Butyl ester (300mg, 0.782mmol), HATU (595mg, 1.564mmol), triethylamine (237mg, 2.346mmol) and dichloromethane were added to the reaction flask (10mL) and reacted at room temperature overnight. After the reaction, the reaction mixture Add dichloromethane (100mL) to dilute, wash with 1N hydrochloric acid aqueous solution (100mL), saturated sodium chloride aqueous solution (100mL) successively, dry with anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure. The ester mixed solvent (9:1, v/v, 50 mL) was slurried, filtered, and dried under reduced pressure to obtain an off-white solid with a yield of 90.5%; ESI-MS (m/z): 709.4 [M+H] + .
步骤f)：(4-(((S)-2-((R)-2-氨基-3-(4-乙氧基苯基)丙氨基)-3-苯丙氨基)甲基)苄基)氨基甲酸叔丁酯的制备Step f): (4-(((S)-2-((R)-2-amino-3-(4-ethoxyphenyl)propylamino)-3-phenylpropylamino)methyl)benzyl ) Preparation of tert-butyl carbamate
将(4-((5R,8S)-8-苄基-5-(4-乙氧基苄基)-3,6,9-三氧代-1-苯基-2-氧-4,7,10-三氮十一烷-11-基)苄基)氨基甲酸叔丁酯(500mg，0.705mmol)和甲醇(70mL)加入反应瓶中，搅拌溶解，加入Pd/C(100mg)，通氢气室温反应4小时，过滤，滤饼用甲醇(50mL)洗涤，合并滤液，减压浓缩，得白色固体，收率89.1％；ESI-MS(m/z)：575.4[M+H] +。 Add (4-((5R,8S)-8-benzyl-5-(4-ethoxybenzyl)-3,6,9-trioxo-1-phenyl-2-oxo-4,7 , 10-Triazaundecane-11-yl)benzyl) tert-butyl carbamate (500mg, 0.705mmol) and methanol (70mL) were added to the reaction flask, stirred to dissolve, Pd/C (100mg) was added, and hydrogen gas was added React at room temperature for 4 hours, filter, wash the filter cake with methanol (50 mL), combine the filtrate, and concentrate under reduced pressure to obtain a white solid with a yield of 89.1%; ESI-MS (m/z): 575.4 [M+H] + .
步骤g)：(4-(((S)-2-((R)-2-苯甲酰氨基-3-(4-乙氧基苯基)丙氨基)-3-苯丙氨基)甲基)苄基)氨基甲酸叔丁酯的制备Step g): (4-(((S)-2-((R)-2-benzamido-3-(4-ethoxyphenyl)propylamino)-3-phenylpropylamino)methyl )Benzyl) preparation of tert-butyl carbamate
将(4-(((S)-2-((R)-2-氨基-3-(4-乙氧基苯基)丙氨基)-3-苯丙氨基)甲基)苄基)氨基甲酸叔丁 酯(361mg，0.628mmol)、三乙胺(191mg，1.884mmol)和二氯甲烷(20mL)加入反应瓶中，搅拌混匀，缓慢滴加苯甲酰氯(106mg，0.754mmol)的二氯甲烷溶液(2mL)中，室温反应6小时，加入二氯甲烷(80mL)稀释，依次用饱和氯化铵(100mL)、饱和食盐水(100mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩至5mL，冷却，过滤析出的固体，滤饼用适量乙酸乙酯洗涤，减压真空干燥，得白色固体，收率57.1％；ESI-MS(m/z)：679.4[M+H] +。步骤h)：N-((R)-1-(((S)-1-((4-(氨甲基)苄基)氨基)-1-氧代-3-苯丙基-2-基)氨基)-3-(4-乙氧基苯基)-1-氧代丙基-2-基)苯甲酰胺的制备 (4-(((S)-2-((R)-2-amino-3-(4-ethoxyphenyl)propylamino)-3-phenylpropylamino)methyl)benzyl)carbamic acid Tert-butyl ester (361mg, 0.628mmol), triethylamine (191mg, 1.884mmol) and dichloromethane (20mL) were added to the reaction flask, stirred and mixed well, and benzoyl chloride (106mg, 0.754mmol) dichloride was slowly added dropwise React in methane solution (2mL) at room temperature for 6 hours, add dichloromethane (80mL) to dilute, wash with saturated ammonium chloride (100mL), saturated brine (100mL), dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure To 5 mL, cool, filter the precipitated solid, wash the filter cake with an appropriate amount of ethyl acetate, and dry under reduced pressure to obtain a white solid with a yield of 57.1%; ESI-MS (m/z): 679.4 [M+H] + . Step h): N-((R)-1-(((S)-1-((4-(aminomethyl)benzyl)amino)-1-oxo-3-phenylpropyl-2-yl )Amino)-3-(4-ethoxyphenyl)-1-oxopropyl-2-yl)benzamide
将(4-(((S)-2-((R)-2-苯甲酰氨基-3-(4-乙氧基苯基)丙氨基)-3-苯丙氨基)甲基)苄基)氨基甲酸叔丁酯(100mg，0.158mmol)和盐酸二氧六环溶液(3mL)加入反应瓶中，室温搅拌反应1.5h，反应结束后，减压蒸干，残余物加乙醇(2mL)超声粉碎打浆，抽滤，滤饼以少量乙醇洗涤，40℃减压真空干燥，得白色固体，收率83.0％， 1H NMR(400MHz,DMSO-d 6)δ8.64(t,J＝6.0Hz,1H),8.59(d,J＝8.4Hz,1H),8.50(d,J＝8.4Hz,1H),8.29(s,3H),7.81-7.65(m,2H),7.51(t,J＝7.6Hz,1H),7.46-7.33(m,4H),7.30-7.08(m,8H),6.76(d,J＝8.4Hz,2H),4.69-4.53(m,2H),4.40-4.21(m,2H),4.04-3.85(m,4H),3.06(dd,J 1＝13.6Hz,J 2＝4.8Hz,1H),2.83(dd,J 1＝13.6,J 2＝10.0Hz,1H),2.69(d,J＝7.6Hz,2H),1.27(t,J＝7.6Hz,3H)；ESI-MS(m/z)：579.3[M+H] +。 Add (4-(((S)-2-((R)-2-benzamido-3-(4-ethoxyphenyl)propylamino)-3-phenylpropylamino)methyl)benzyl ) Tert-butyl carbamate (100mg, 0.158mmol) and dioxane hydrochloride solution (3mL) were added to the reaction flask, stirred at room temperature for 1.5h, after the reaction, evaporated to dryness under reduced pressure, the residue was added ethanol (2mL) ultrasound Crushing and beating, suction filtration, the filter cake is washed with a small amount of ethanol, and dried under reduced pressure at 40°C to obtain a white solid with a yield of 83.0%. 1 H NMR (400MHz, DMSO-d 6 )δ8.64(t,J=6.0Hz ,1H),8.59(d,J=8.4Hz,1H),8.50(d,J=8.4Hz,1H),8.29(s,3H),7.81-7.65(m,2H),7.51(t,J= 7.6Hz, 1H), 7.46-7.33 (m, 4H), 7.30-7.08 (m, 8H), 6.76 (d, J = 8.4Hz, 2H), 4.69-4.53 (m, 2H), 4.40-4.21 (m ,2H),4.04-3.85(m,4H),3.06(dd,J 1 = 13.6Hz, J 2 = 4.8Hz, 1H), 2.83(dd, J 1 = 13.6, J 2 = 10.0Hz, 1H), 2.69 (d, J=7.6 Hz, 2H), 1.27 (t, J=7.6 Hz, 3H); ESI-MS (m/z): 579.3 [M+H] + .
实施例74 生物活性测试Example 74 Biological Activity Test
1.人血浆激肽释放酶(Human PK)抑制活性测定1. Determination of the inhibitory activity of human plasma kallikrein (Human PK)
采用Johansen等(Johansen el al.,Int.J.Tiss.Reac.1986,8,185)报道的方法，并做出部分改进，反应液为10mM PBS,1mM EDTA,0.1％BSA,pH＝7.4。向384微孔板中依次加入0.4nM人血浆激肽释放酶(商购自Enzyme Research Laboratories)10μL，待测化合物溶液5μL，加毕混匀后，37℃孵育15min，向各孔加入5μL加反应底物N-Benzoyl-pro-phe-Arg-p-Nitroanilide(商购自sigma)(500μM)显色，用酶标仪动力学模式测定405nm处各孔的光密度(OD值)，与不加待测样品的空白孔比较，计算化合物对酶的抑制率[抑制率＝(1-样品组OD值/空白组OD值)x100％]，在Prism GraphPad中用四参数模式计算IC 50值；每个化合物每次测定2个复孔，每组实验独立重复三次。测试数据如表1所示： Using the method reported by Johansen et al. (Johansen el al., Int. J. Tiss. Reac. 1986, 8, 185), and making some improvements, the reaction solution is 10 mM PBS, 1 mM EDTA, 0.1% BSA, and pH=7.4. Add 10μL of 0.4nM human plasma kallikrein (commercially available from Enzyme Research Laboratories) and 5μL of the test compound solution to the 384 microwell plate. After adding and mixing, incubate at 37°C for 15min, and add 5μL to each well for reaction The substrate N-Benzoyl-pro-phe-Arg-p-Nitroanilide (commercially available from sigma) (500μM) develops the color, and the optical density (OD value) of each well at 405nm is measured by the kinetic mode of the microplate reader. Compare the blank wells of the sample to be tested, calculate the inhibitory rate of the compound against the enzyme [inhibition rate = (1-sample group OD value/blank group OD value) x 100%], use the four-parameter mode to calculate the IC 50 value in Prism GraphPad; Each compound was tested in 2 replicate wells, and each experiment was repeated three times independently. The test data is shown in Table 1:
表1Table 1
化合物编号Compound number IC50(人血浆激肽释放酶)nMIC50 (human plasma kallikrein) nM
化合物1Compound 1 3.13.1
化合物2 Compound 2 12.812.8
化合物3 Compound 3 9.49.4
化合物4Compound 4 8.88.8
化合物5Compound 5 3.33.3
化合物6Compound 6 0.980.98
化合物7Compound 7 9.99.9
化合物8Compound 8 9.19.1
化合物9Compound 9 48.648.6
化合物10 Compound 10 10.010.0
化合物11Compound 11 384.5384.5
化合物12Compound 12 42.842.8
化合物13Compound 13 6.96.9
化合物14Compound 14 25.625.6
化合物15 Compound 15 4.94.9
化合物16Compound 16 7.67.6
化合物17Compound 17 78.578.5
化合物18Compound 18 8.68.6
化合物19Compound 19 98.798.7
化合物20Compound 20 285.1285.1
化合物21Compound 21 178.6178.6
化合物22Compound 22 0.990.99
化合物23Compound 23 1.81.8
化合物24Compound 24 2.32.3
化合物25Compound 25 3.23.2
化合物26Compound 26 6.06.0
化合物27Compound 27 312.4312.4
化合物28Compound 28 85.185.1
化合物29Compound 29 62.862.8
化合物30Compound 30 0.680.68
化合物31Compound 31 3.93.9
化合物32Compound 32 0.910.91
化合物33Compound 33 2.52.5
化合物34Compound 34 16.516.5
化合物35Compound 35 3.93.9
化合物36Compound 36 4.84.8
化合物37Compound 37 5.75.7
化合物38Compound 38 302.3302.3
化合物39Compound 39 476.5476.5
化合物40Compound 40 8.98.9
化合物41Compound 41 54.354.3
化合物42Compound 42 69.769.7
化合物43Compound 43 200.6200.6
化合物44Compound 44 35.435.4
化合物45Compound 45 20.020.0
化合物46Compound 46 60.560.5
化合物47Compound 47 110.9110.9
化合物48Compound 48 47.847.8
化合物49Compound 49 98.698.6
化合物50 Compound 50 369.5369.5
化合物51Compound 51 83.583.5
化合物52Compound 52 254.1254.1
化合物53Compound 53 0.990.99
化合物54Compound 54 1.011.01
化合物55Compound 55 0.790.79
化合物56Compound 56 11.811.8
化合物57Compound 57 18.818.8
化合物58Compound 58 69.969.9
化合物59Compound 59 96.596.5
化合物60 Compound 60 459.3459.3
化合物61Compound 61 405.3405.3
化合物62Compound 62 278.5278.5
化合物63Compound 63 1.91.9
化合物64Compound 64 4.04.0
化合物65Compound 65 3.33.3
化合物66Compound 66 28.728.7
化合物67Compound 67 32.832.8
化合物68Compound 68 401.7401.7
化合物69Compound 69 66.666.6
化合物70 Compound 70 55.755.7
化合物71Compound 71 2.02.0
化合物72Compound 72 48.348.3
化合物73Compound 73 152.5152.5
2.人凝血酶(thrombin)抑制活性测定2. Determination of human thrombin (thrombin) inhibitory activity
采用H.C.Hemker等(Handbook of Synthetic Substrates)报道的方法，并做出部分改进，反应液为50mM Tris-HCl,pH＝8.3,130mM NaCl,0.5％BSA。向384微孔板中依次加入0.2nM人凝血酶(商购自Enzyme Research Laboratories)10μL，待测化合物溶液5μL，加毕混匀后，37℃毕孵育15min，向各孔加入5μL 75μM反应底物BOC-Val-Pro-AFC(商购自sigma)显色，用酶标仪动力学模式于380nm激发波长，500nm发射波长处测定各孔的荧光值，与不加待测样品的空白孔比较，计算化合物对酶的抑制率[抑制率＝(1-样品组荧光值/空白组荧光值)x100％]，在Prism GraphPad中用四参数模式计算IC 50值；每个化合物每次测定2个复孔，每组实验独立重复三次。 The method reported by HCHemker et al. (Handbook of Synthetic Substrates) was used with some improvements. The reaction solution was 50mM Tris-HCl, pH=8.3, 130mM NaCl, 0.5% BSA. Add 10μL of 0.2nM human thrombin (commercially available from Enzyme Research Laboratories) and 5μL of the test compound solution to the 384 microwell plate. After adding and mixing, incubate at 37°C for 15min, and add 5μL of 75μM reaction substrate to each well BOC-Val-Pro-AFC (commercially available from sigma) is used for color development, and the fluorescence value of each well is measured at the excitation wavelength of 380nm and emission wavelength of 500nm in the kinetic mode of the microplate reader. Compared with the blank well without the sample to be tested, Calculate the inhibition rate of the compound against the enzyme [inhibition rate = (1-sample group fluorescence value/blank group fluorescence value) x 100%], use the four-parameter mode in Prism GraphPad to calculate the IC 50 value; each compound measures 2 replicates each time Well, each experiment was repeated three times independently.
3.胰蛋白酶(Trypsin)抑制活性测定3. Determination of trypsin inhibitory activity
采用H.C.Hemker等(Handbook of Synthetic Substrates)报道的方法，并做出部分改进，反应液为200mM Tris-HCl,20mM CaCl 2，pH＝7.8。向384微孔板中依次加入12.5nM胰蛋 白酶(商购自Biovision)10μL，待测化合物溶液5μL，加毕混匀后，37℃孵育15min，向各孔加入5μL 200μM L-BAPA(商购自sigma)显色，用酶标仪动力学模式测定405nm处各孔的光密度(OD值)，与不加待测样品的空白孔比较，计算化合物对酶的抑制率[抑制率＝(1-样品组OD值/空白组OD值)x100％]，在Prism GraphPad中用四参数模式计算IC 50值；每个化合物每次测定2个复孔，每组实验独立重复三次。 Using the method reported by HCHemker et al. (Handbook of Synthetic Substrates), and making some improvements, the reaction solution is 200mM Tris-HCl, 20mM CaCl 2 , pH=7.8. Add 10μL of 12.5nM trypsin (commercially available from Biovision) to the 384 microwell plate, and 5μL of the test compound solution. After adding and mixing, incubate at 37°C for 15min. Add 5μL of 200μM L-BAPA (commercially available from sigma), use the kinetic mode of the microplate reader to measure the optical density (OD value) of each well at 405nm, compare with the blank well without the sample to be tested, and calculate the inhibitory rate of the compound against the enzyme [inhibition rate = (1- Sample group OD value/blank group OD value) x 100%], the IC 50 value was calculated in the Prism GraphPad using a four-parameter mode; each compound was measured in 2 replicate wells each time, and each group of experiments was independently repeated three times.
4.人凝血因子XIa(Factor XIa)抑制活性测定4. Determination of human coagulation factor XIa (Factor XIa) inhibitory activity
采用H.C.Hemker等(Handbook_of_Synthetic_Subst)报道的方法，并做出部分改进，反应液为0.05M Tris/HCl,0.15M NaCl，BSA(0.1mg/ml)，pH＝8.0。向384微孔板中依次加入15nM人凝血因子XIa(商购自Enzyme Research Laboratories)10μL，待测化合物溶液5μL，加毕混匀后，37℃孵育15min，向各孔加入5μL 400μM反应底物N-Benzoyl-pro-phe-Arg-p-Nitroanilide(商购自sigma)显色，用酶标仪动力学模式测定405nm处各孔的光密度(OD值)，与不加待测样品的空白孔比较，计算化合物对酶的抑制率[抑制率＝(1-样品组OD值/空白组OD值)x100％]，在Prism GraphPad中用四参数模式计算IC 50值；每个化合物每次测定2个复孔，每组实验独立重复三次。 The method reported by HCHemker et al. (Handbook_of_Synthetic_Subst) was used with some improvements. The reaction solution was 0.05M Tris/HCl, 0.15M NaCl, BSA (0.1mg/ml), and pH=8.0. Add 10μL of 15nM human coagulation factor XIa (commercially available from Enzyme Research Laboratories) and 5μL of the test compound solution to the 384 microwell plate. After adding and mixing, incubate at 37°C for 15min, and add 5μL of 400μM reaction substrate N to each well. -Benzoyl-pro-phe-Arg-p-Nitroanilide (commercially available from sigma) develops the color, and the optical density (OD value) of each hole at 405nm is measured by the kinetic mode of the microplate reader, and the blank hole without the sample to be tested Compare and calculate the inhibition rate of the compound against the enzyme [inhibition rate=(1-sample group OD value/blank group OD value) x 100%], use the four-parameter mode to calculate the IC 50 value in Prism GraphPad; each compound is measured 2 times each time Each set of experiments was repeated three times independently.
5.人凝血因子Xa(Factor Xa)抑制活性测定5. Determination of human coagulation factor Xa (Factor Xa) inhibitory activity
采用H.C.Hemker等(Handbook of Synthetic Substrates)报道的方法，并做出部分改进，反应液为0.05M Tris,0.1M NaCl，pH＝7.4。向384微孔板中依次加入3nM人凝血因子Xa(商购自Enzyme Research Laboratories)10μL，待测化合物溶液5μL，加毕混匀后，37℃孵育15min，向各孔加入5μL 400μM反应底物N-Benzoyl-Val-Gly-Arg p-nitroanilide hydrochloride显色，用酶标仪动力学模式测定405nm处各孔的光密度(OD值)，与不加待测样品的空白孔比较，计算化合物对酶的抑制率[抑制率＝(1-样品组OD值/空白组OD值)x100％]，在Prism GraphPad中用四参数模式计算IC 50值；每个化合物每次测定2个复孔，每组实验独立重复三次。 The method reported by HCHemker et al. (Handbook of Synthetic Substrates) was adopted, and some improvements were made. The reaction solution was 0.05M Tris, 0.1M NaCl, and pH=7.4. Add 10μL of 3nM human coagulation factor Xa (commercially available from Enzyme Research Laboratories) and 5μL of the test compound solution to the 384 microwell plate. After adding and mixing, incubate at 37°C for 15min, and add 5μL of 400μM reaction substrate N to each well. -Benzoyl-Val-Gly-Arg p-nitroanilide hydrochloride color development, use the microplate reader kinetic mode to measure the optical density (OD value) of each hole at 405nm, compare with the blank hole without the test sample, calculate the compound to enzyme Inhibition rate [inhibition rate = (1-sample group OD value/blank group OD value) x 100%], the IC 50 value is calculated in Prism GraphPad with four-parameter mode; each compound is measured in 2 replicate wells, each group The experiment was repeated three times independently.
6.人血纤维蛋白溶酶(plasmin)抑制活性测定6. Determination of human plasmin inhibitory activity
采用H.C.Hemker等(Handbook of Synthetic Substrates)报道的方法，并做出部分改进，反应液为0.05M Tris/HC1,0.13M NaCl，BSA(5mg/ml)，pH＝8.3。向384微孔板中依次加入2nM人血纤维蛋白溶酶(商购自Enzyme Research Laboratories)10μL，待测化合物溶液5μL，加毕混匀后，37℃孵育15min，向各孔加入5μL 1.2mM反应底物Tosyl-Gly-Pro-Lys-4-nitranilide(商购自Sigma)显色，用酶标仪动力学模式测定405nm处各孔的光密度(OD值)，与不加待测样品的空白孔比较，计算化合物对酶的抑制率[抑制率＝(1-样品组OD值/空白组OD值)x100％]，在Prism GraphPad中用四参数模式计算IC 50值；每个化合物每次测定2个复孔，每组实验独立重复三次。 The method reported by HCHemker et al. (Handbook of Synthetic Substrates) was used with some improvements. The reaction solution was 0.05M Tris/HC1, 0.13M NaCl, BSA (5mg/ml), and pH=8.3. Add 10μL of 2nM human plasmin (commercially available from Enzyme Research Laboratories) and 5μL of the test compound solution to the 384 microwell plate. After adding and mixing, incubate at 37°C for 15min, and add 5μL of 1.2mM to each well. The substrate Tosyl-Gly-Pro-Lys-4-nitranilide (commercially available from Sigma) develops the color, and the optical density (OD value) of each well at 405nm is measured in the kinetic mode of the microplate reader, and the blank without adding the sample to be tested Comparing the wells, calculate the inhibition rate of the compound against the enzyme [inhibition rate = (1-sample group OD value/blank group OD value) x 100%], use the four-parameter mode in Prism GraphPad to calculate the IC 50 value; each compound is measured every time 2 duplicate wells, and each experiment was repeated three times independently.
上述人凝血酶、胰蛋白酶、人凝血因子XIa、人凝血因子Xa、人血纤维蛋白溶酶抑制活性测试数据如表2：The above-mentioned human thrombin, trypsin, human coagulation factor XIa, human coagulation factor Xa, human plasmin inhibitory activity test data are shown in Table 2:
表2Table 2
化合物编号Compound number 人凝血酶Human thrombin 胰蛋白酶Trypsin 凝血因子XIaCoagulation factor XIa 凝血因子XaCoagulation factor Xa 血纤维蛋白溶酶Plasmin
化合物1Compound 1 >10μM>10μM >10μM>10μM >10μM>10μM 4.8μM4.8μM >10μM>10μM
化合物2Compound 2 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物3Compound 3 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物5Compound 5 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物6Compound 6 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物13Compound 13 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物15Compound 15 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物22Compound 22 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物23Compound 23 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物24Compound 24 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物30Compound 30 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物31Compound 31 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物32Compound 32 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物33Compound 33 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物34Compound 34 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物35Compound 35 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物41Compound 41 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物45Compound 45 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物46Compound 46 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物53Compound 53 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物54Compound 54 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物63Compound 63 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物64Compound 64 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物71Compound 71 >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM >10μM>10μM
化合物73Compound 73 >10μM>10μM 10.8μM10.8μM >10μM>10μM >10μM>10μM >10μM>10μM
实施例75 化合物的溶解度测试Example 75 Compound solubility test
将10μL化合物1、化合物6、化合物22、化合物30、化合物53、化合物71、化合物73的DMSO溶液(10mM)分别添加到whatman miniuniprep小瓶的下腔室中，然后在whatman miniuniprep小瓶的下腔室中分别添加490μL的50mM PB(磷酸盐缓冲液，将15mL 50mM Na 2HPO 4添加到50mL管中，然后用50mM NaH 2PO 4将pH调节至7.4)。将样品涡旋至少2分钟，在室温下以800rpm的速度将miniuniprep小瓶在barnstead摇床上摇动24小时，之后离心20分钟(例如4000rpm)，压缩的miniunipreps制备的滤液注入HPLC系统，色谱柱：Waters XBridge C18 4.6*100mm，流动相A：0.1％TFA的乙腈/水溶液(5:95)，流动相B：0.1％TFA的乙腈/水溶液(95:5)，流速1.4mL/min，检测波长305nm，进样量5μL，梯度洗脱条件如表3： Add 10 μL of compound 1, compound 6, compound 22, compound 30, compound 53, compound 71, compound 73 DMSO solution (10 mM) into the lower chamber of the whatman miniuniprep vial, and then in the lower chamber of the whatman miniuniprep vial Add 490 μL of 50 mM PB (phosphate buffer, 15 mL of 50 mM Na 2 HPO 4 into a 50 mL tube, and then adjust the pH to 7.4 with 50 mM NaH 2 PO 4 ). Vortex the sample for at least 2 minutes, shake the miniuniprep vial on a barnstead shaker for 24 hours at room temperature at 800 rpm, then centrifuge for 20 minutes (for example, 4000 rpm), and inject the filtrate prepared by compressed miniunipreps into the HPLC system. Column: Waters XBridge C18 4.6*100mm, mobile phase A: 0.1% TFA acetonitrile/water solution (5:95), mobile phase B: 0.1% TFA acetonitrile/water solution (95:5), flow rate 1.4mL/min, detection wavelength 305nm, enter The sample volume is 5μL, and the gradient elution conditions are shown in Table 3:
表3table 3
时间(min)Time (min) A(％)A(%) B(％)B(%)
0.010.01 100100 00
4.004.00 5050 5050
4.014.01 100100 00
5.005.00 100100 00
以标准曲线计算浓度，结果如下表4所示：Calculate the concentration with the standard curve, and the results are shown in Table 4 below:
表4Table 4
编号Numbering 动力学溶解度(μM)Kinetic solubility (μM)
化合物1 Compound 1 45.045.0
化合物6Compound 6 29.229.2
化合物22Compound 22 79.479.4
化合物30Compound 30 17.017.0
化合物53Compound 53 3.363.36
化合物71Compound 71 36.236.2
化合物73Compound 73 12.012.0
实施例76 人肝微粒体稳定性测试Example 76 Stability Test of Human Liver Microsomes
1.材料1. Material
1.1肝微粒体1.1 Liver microsomes
人和动物微粒体购买于Corning或Xenotech，储存于-80℃冰箱。Human and animal microsomes were purchased from Corning or Xenotech and stored in a refrigerator at -80°C.
1.2还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)，供应商：Chem-impex international，1.2 Reduced nicotinamide adenine dinucleotide phosphate (NADPH), supplier: Chem-impex international,
1.3对照化合物：睾酮，双氯芬酸，普罗帕酮1.3 Control compounds: testosterone, diclofenac, propafenone
1.4主要仪器质谱仪Sciex API4000；高效液相色谱LC20-AD，Shimadzu；进样器ADDA，Apricot Designs1.4 Main instruments: Mass spectrometer Sciex API4000; High performance liquid chromatography LC20-AD, Shimadzu; Sample injector ADDA, Apricot Designs
2.实验步骤2. Experimental steps
2.1供试品溶液的配制2.1 Preparation of test solution
称取实施例化合物适量，加入DMSO溶解为10mM的母液；取适量体积母液，100％乙腈稀释到100μM。Weigh an appropriate amount of the compound of the embodiment, add DMSO to dissolve the mother liquor to 10mM; take an appropriate volume of the mother liquor, and dilute it to 100μM with 100% acetonitrile.
2.2实验步骤2.2 Experimental steps
准备2块96孔孵育板，分别命名为T60孵育板和NCF60孵育板。在T60孵育板和NCF60孵育板上分别加入445μL微粒体工作液(肝微粒体蛋白浓度为0.56mg/mL)，然后将上述孵育板放置于37℃水浴锅中预孵育大约10分钟。Prepare two 96-well incubation plates, named T60 incubation plate and NCF60 incubation plate, respectively. Add 445 μL of microsomal working solution (liver microsomal protein concentration of 0.56 mg/mL) to the T60 incubation plate and NCF60 incubation plate respectively, and then place the incubation plate in a 37°C water bath for pre-incubation for about 10 minutes.
预孵育结束后，在T60孵育板和NCF60孵育板上分别加入5μL供试品或对照化合物溶液，混匀。在NCF60孵育板上每孔添加50μL磷酸钾盐缓冲液启动反应；在T0终止板中加入180μL的终止液(含200ng/mL tolbutamide和200ng/mL labetalol的乙腈溶液)和6uL的NADPH再生体系工作液，从T60孵育板中取出54μL样品至T0终止板(T0样品产生)。在T60孵育板上每孔添加44μLNADPH再生体系工作液启动反应。在Blank板中只添加54μL微粒体样品溶液、6uL的NADPH再生体系工作液和180μL的终止液。因此，在供试品或对照化合物的样品中，化合物、睾酮、双氯芬酸和普罗帕酮的反应终浓度为1μM，肝微粒体的浓度为0.5mg/mL，DMSO和乙腈在反应体系中的终浓度分别为0.01％(v/v)和0.99％(v/v)。孵育适当时间(如5、10、20、30和60分钟)后，分别在每个终止板的样品孔中加入180μL的终止液(含200ng/mL tolbutamide和200ng/mL labetalol的乙腈溶液)，之后从T60孵育板中取出60μL样品以终止反应。所有样品板摇匀并在3220×g离心20分钟，然后每孔取80μL上清液稀释到240μL纯水中用于液相色谱串联质谱分析。After the pre-incubation, add 5 μL of the test substance or control compound solution to the T60 incubation plate and the NCF60 incubation plate, and mix well. Add 50μL of potassium phosphate buffer to each well of the NCF60 incubation plate to start the reaction; add 180μL of stop solution (containing 200ng/mL tolbutamide and 200ng/mL labetalol in acetonitrile solution) and 6uL of NADPH regeneration system working solution to the T0 stop plate , Take 54μL sample from T60 incubation plate to T0 stop plate (T0 sample generation). Add 44μL of NADPH regeneration system working solution to each well of the T60 incubation plate to start the reaction. Only 54μL of microsome sample solution, 6uL of NADPH regeneration system working solution and 180μL of stop solution were added to the Blank plate. Therefore, in the sample of the test product or control compound, the final concentration of the compound, testosterone, diclofenac and propafenone in the reaction is 1μM, the concentration of liver microsomes is 0.5mg/mL, and the final concentration of DMSO and acetonitrile in the reaction system They are 0.01% (v/v) and 0.99% (v/v), respectively. After incubating for an appropriate time (such as 5, 10, 20, 30, and 60 minutes), add 180 μL of stop solution (acetonitrile solution containing 200ng/mL tolbutamide and 200ng/mL labetalol) into the sample wells of each stop plate, and then Take out 60 μL of sample from the T60 incubation plate to stop the reaction. All sample plates were shaken well and centrifuged at 3220×g for 20 minutes, and then 80 μL of supernatant was diluted into 240 μL of pure water from each well for liquid chromatography tandem mass spectrometry analysis.
3.液相色谱串联质谱对所有样品进样分析，以体外代谢半衰期(T 1/2,min)和固有清除率(CL int)作为指标，考察化合物的代谢稳定性，结果如表5所示： 3. Liquid chromatography tandem mass spectrometry analyzes all samples, using in vitro metabolic half-life (T 1/2 , min) and intrinsic clearance (CL int ) as indicators to investigate the metabolic stability of the compounds. The results are shown in Table 5. :
表5table 5
Figure PCTCN2020137640-appb-000324
Figure PCTCN2020137640-appb-000324
实施例77 细胞色素P450同工酶抑制性实验Example 77 Cytochrome P450 isoenzyme inhibition experiment
1.材料1. Material
1.1细胞色素P450同工酶1.1 Cytochrome P450 isoenzymes
人肝微粒体细胞色素P450同工酶CYP1A2、CYP2C9、CYP2C19、CYP2D6和CYP3A4。Human liver microsomal cytochrome P450 isoenzymes CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4.
1.2阳性抑制剂1.2 Positive inhibitors
α-萘黄酮、磺胺苯吡唑、(S)-(+)-N-3-苄基苯乙基内酰脲、奎尼丁、酮康唑α-Naphthoflavone, Sulfapyrazole, (S)-(+)-N-3-benzylphenethyl lactam, Quinidine, Ketoconazole
1.3特异性底物1.3 Specific substrate
非那西丁、双氯芬酸、(S)-美芬妥英、右美沙芬、咪哒唑仑Phenacetin, Diclofenac, (S)-Mephenytoin, Dextromethorphan, Midazolam
1.4主要仪器1.4 Main instruments
质谱仪Sciex API4000TM；超高效液相色谱LC30-AD，Shimadzu；自动进样器SIL30-ACMP,ShimadzuMass spectrometer Sciex API4000TM; ultra high performance liquid chromatography LC30-AD, Shimadzu; autosampler SIL30-ACMP, Shimadzu
2.实验步骤2. Experimental steps
2.1工作液的配制2.1 Preparation of working fluid
储备液：受试化合物采用DMSO溶解，制备成10mM DMSO储备液；Stock solution: The test compound is dissolved in DMSO and prepared into a 10mM DMSO stock solution;
工作浓度配制：将适量的K 2HPO 4·3H 2O(分析纯)和KH 2PO 4(分析纯)溶解在适量的超纯水中，用H 3PO 4或KOH将pH调至7.40±0.10，配成浓度为100mM的缓冲液，即得磷酸钾缓冲液(PB)。 Working concentration preparation: Dissolve an appropriate amount of K 2 HPO 4 ·3H 2 O (analytical purity) and KH 2 PO 4 (analytical purity) in an appropriate amount of ultrapure water, and adjust the pH to 7.40± with H 3 PO 4 or KOH 0.10, formulated into a buffer with a concentration of 100 mM, to obtain potassium phosphate buffer (PB).
2.2实验步骤2.2 Experimental steps
(1)首先将受试化合物储备液(10.0mM)用甲醇(分析纯)稀释至1mM备用；(1) First, dilute the test compound stock solution (10.0mM) with methanol (analytical purity) to 1mM for later use;
(2)同时准备P450同工酶(CYP1A2、CYP2C9、CYP2C19、CYP2D6、CYP3A4)混合物(5合1)的工作液：将保存在低于-60℃冰箱的人肝微粒体置于冰上解冻，待人肝微粒体全部溶解，用PB进行稀释，制备一定浓度工作液(0.253mg/mL)；各阳性抑制剂配成0.3mM备用；(2) At the same time, prepare the working solution of the mixture of P450 isoenzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4) (5 in 1): Thaw human liver microsomes stored in a refrigerator below -60 ℃ on ice, After the human liver microsomes are completely dissolved, dilute with PB to prepare a working solution of a certain concentration (0.253mg/mL); each positive inhibitor is made up to 0.3mM for use;
(3)先将20.0μL底物混合液加至反应板中(Blank孔中加入20.0μL PB)，然后将158μL人肝微粒体工作液加入反应板中，将反应板置于冰上，待用；此时将2.00μL受试化合物(N＝2)及特异性抑制剂(N＝2)加入对应孔中，无抑制剂(受试化合物或阳性抑制剂)组加入对应的有机溶剂，作为对照组样品(受试化合物对照样品为1:1DMSO:MeOH，阳性对照样品为1:9DMSO:MeOH)；(3) First add 20.0μL of substrate mixture to the reaction plate (add 20.0μL of PB to the Blank well), then add 158μL of human liver microsome working solution to the reaction plate, place the reaction plate on ice, and set aside ; At this time, add 2.00 μL of the test compound (N=2) and specific inhibitor (N=2) to the corresponding wells, and add the corresponding organic solvent to the group without inhibitor (test compound or positive inhibitor) as a control Group samples (the test compound control sample is 1:1DMSO:MeOH, and the positive control sample is 1:9DMSO:MeOH);
(4)在37℃水浴预孵育10min后，将20.0μL辅酶因子(NADPH)溶液加入反应板中，置于37℃水浴孵育反应10min；(4) After pre-incubating in a 37°C water bath for 10 minutes, add 20.0 μL of Coenzyme Factor (NADPH) solution to the reaction plate and place it in a 37°C water bath to incubate the reaction for 10 minutes;
(5)加入400μL预冷的乙腈溶液(含200ng/mL Tolbutamide和Labetalol的内标)终止反应；(5) Add 400μL of pre-cooled acetonitrile solution (containing 200ng/mL Tolbutamide and Labetalol internal standard) to stop the reaction;
(6)将反应板置于摇床，振荡10min混匀；然后在4℃、4000rpm条件下离心20min；取200μL上清加至100μL水中，进行样品稀释；最后封板，振荡10min混匀，进行液相色谱串联质谱检测。(6) Place the reaction plate on a shaker and shake for 10 minutes to mix; then centrifuge at 4°C and 4000 rpm for 20 minutes; add 200 μL of supernatant to 100 μL of water to dilute the sample; finally seal the plate, shake for 10 minutes, and mix well. Liquid chromatography tandem mass spectrometry detection.
3.液相色谱串联质谱分析3. Liquid chromatography tandem mass spectrometry analysis
所有样品进样分析。以受试化合物对人肝微粒体细胞色素P450同工酶的抑制率作为指标，考察受试化合物对肝脏代谢酶的影响，结果表明化合物6和化合物22对五种重要CYP酶均无明显抑制，IC50大于10μM，具体结果如表6：All samples are injected and analyzed. The inhibitory rate of the test compound on human liver microsomal cytochrome P450 isoenzymes was used as an index to investigate the effect of the test compound on liver metabolic enzymes. The results showed that compound 6 and compound 22 did not significantly inhibit the five important CYP enzymes. IC50 is greater than 10μM, the specific results are shown in Table 6:
表6 10μM受试化合物对P450的抑制率Table 6 Inhibition rate of 10μM test compound on P450
Figure PCTCN2020137640-appb-000325
Figure PCTCN2020137640-appb-000325
实施例78 碳酸酐酶I诱导的视网膜血管通透性模型Example 78 Carbonic anhydrase I-induced retinal vascular permeability model
1、实验系统1. Experimental system
种属：SD大鼠(购自成都达硕实验动物有限公司)；Species: SD rats (purchased from Chengdu Dashuo Experimental Animal Co., Ltd.);
等级：SPF级；Grade: SPF grade;
使用动物数量和性别：雄性，9只/组(共6组，合计54只)；Number and sex of animals used: male, 9 per group (6 groups in total, 54 in total);
年龄：购入时年龄6-8周龄；Age: 6-8 weeks old at the time of purchase;
购买时体重：250±20g；Weight at the time of purchase: 250±20g;
2、样品溶液配制2. Sample solution preparation
称取实施例化合物适量，加入DMSO溶解为10mM的母液；取适量母液，加入一定体积的超纯水进行稀释，配制为100μM工作液；再取适量工作液，加入一定体积生理盐水进行稀释，配制为0.25μM或2.5μM的最终工作液作为供试品溶液。Weigh an appropriate amount of the compound of the example and add DMSO to dissolve the mother liquor into 10mM; take an appropriate amount of mother liquor and add a certain volume of ultrapure water to dilute to prepare a 100μM working solution; then take an appropriate amount of working solution and add a certain volume of physiological saline to dilute and prepare The final working solution of 0.25μM or 2.5μM is used as the test solution.
3、动物分组3. Animal grouping
组别设计：空白对照组(生理盐水+生理盐水Sham group)、模型对照组(生理盐水+CA-Ⅰ group)、供试品组(化合物22、6、71、73，共计6组)；其中，CA-I为大鼠碳酸酐酶1(购自Sigma-Aldrich)。Group design: blank control group (normal saline + normal saline Sham group), model control group (normal saline + CA-I group), test product group (compounds 22, 6, 71, 73, a total of 6 groups); , CA-I is rat carbonic anhydrase 1 (purchased from Sigma-Aldrich).
4、模型制作4. Model making
复方托吡卡胺滴眼液散瞳，使用1％戊巴比妥钠腹腔注射全身麻醉，显微镜下固定大鼠于操作台上，使用妥布霉素地塞米松滴眼液冲洗结膜囊，聚角巩膜缘1mm处垂直进针，玻璃体注射2uL生理盐水或供试品化合物(化合物22、6、71、73)，30min后，玻璃体内第二次注射2uL生理盐水或CA-I(60ng/眼)造模(单眼)。Compound tropikamide eye drops were used to dilate pupils, and 1% pentobarbital sodium was used for intraperitoneal injection of general anesthesia. The rats were fixed on the operating table under a microscope, and tobramycin and dexamethasone eye drops were used to flush the conjunctival sac. The needle was inserted vertically at 1mm of the corneoscleral limbus, and 2uL normal saline or test compound (compounds 22, 6, 71, 73) was injected into the vitreous body. 30 minutes later, 2uL normal saline or CA-I (60ng/eye) was injected into the vitreous for the second time. ) Modeling (monocular).
5、眼底血管造影观察5. Observation of fundus angiography
检查方法：所有入组动物在第二次玻璃体内注射后30min腹腔注射10％荧光素钠注射液1.5ml/kg，10-15min后行荧光造影，观察眼底视网膜血管渗漏情况，并对渗漏情况进行评分。0：无渗漏；1：轻微渗漏；2：中度渗漏；3：剧烈/重度渗漏。Inspection method: All the animals in the group were intraperitoneally injected with 10% fluorescein sodium injection 1.5ml/kg 30min after the second intravitreal injection, and fluorescein angiography was performed after 10-15min to observe the leakage of the retinal blood vessels in the fundus and the leakage The situation is scored. 0: No leakage; 1: Slight leakage; 2: Moderate leakage; 3: Severe/severe leakage.
6、数据分析6. Data analysis
各组动物实验定量数据采用均数±标准误(
Figure PCTCN2020137640-appb-000326
E M)描述。正态和方差齐的多组间比较采用单因素方差分析(One-Way ANOVA)，组间比较采用方差分析LSD法检验；不满足正态分布或者方差不齐，采用Kruskal-Wallis H检验(K-W法)进行分析，组间多重比较采用Mann-Whitney U方法。P<0.05为差异具有统计学意义。所有的统计分析，均使用SPSS 24软件完成。 The quantitative data of each group of animal experiments adopts the mean ± standard error (
Figure PCTCN2020137640-appb-000326
E M) description. One-way analysis of variance (One-Way ANOVA) was used for multi-group comparisons of normal and homogeneous variances, and the analysis of variance LSD test was used for comparisons between groups; the Kruskal-Wallis H test (KW Method) for analysis, and Mann-Whitney U method was used for multiple comparisons between groups. P<0.05 indicates that the difference is statistically significant. All statistical analysis is done using SPSS 24 software.
结果如图1所示。The result is shown in Figure 1.
实施例79 碳酸酐酶I诱导的视网膜血管通透性模型Example 79 Carbonic anhydrase I-induced retinal vascular permeability model
1、实验系统1. Experimental system
种属：SD大鼠(购自浙江维通利华实验动物技术有限公司)；Species: SD rats (purchased from Zhejiang Weitong Lihua Laboratory Animal Technology Co., Ltd.);
等级：SPF级；Grade: SPF grade;
使用动物数量和性别：雄性，4只/组(共6组，合计24只)；Number and sex of animals used: male, 4 per group (6 groups in total, 24 in total);
年龄：购入时年龄56-69天；Age: 56-69 days old at the time of purchase;
购买时体重：252.73-391.52g；Weight at the time of purchase: 252.73-391.52g;
2、样品溶液配制2. Sample solution preparation
称取实验化合物适量，加入DMSO溶解为10mM的母液；取适量母液，加入一定体积的超纯水进行稀释，配制为100μM工作液；取适量工作液，加入一定体积生理盐水进行稀释，配制为0.1μM或1μM的最终工作液作为供试品组。Weigh an appropriate amount of the experimental compound and add DMSO to dissolve the mother liquor to 10mM; take an appropriate amount of mother liquor and add a certain volume of ultrapure water to dilute to prepare a 100μM working solution; take an appropriate amount of working solution and add a certain volume of physiological saline to dilute it to prepare 0.1 The final working solution of μM or 1 μM is used as the test product group.
3、动物分组3. Animal grouping
组别设计：空白对照组(生理盐水+生理盐水Sham group)、模型对照组(生理盐水+CA-Ⅰ group)、供试品组(化合物1、6、22、73，共计6组)；其中，CA-I为大鼠碳酸酐酶1(购自Sigma-Aldrich)。Group design: blank control group (normal saline + normal saline Sham group), model control group (normal saline + CA-I group), test product group (compounds 1, 6, 22, 73, a total of 6 groups); , CA-I is rat carbonic anhydrase 1 (purchased from Sigma-Aldrich).
4、模型制作4. Model making
以5～15mg/kg和2～12mg/kg的剂量肌肉注射舒泰(Zoletil)和甲苯噻嗪(Xylazine)麻醉动物，同时将奥昔卡因(Oxybuprocaine)滴眼液滴入结膜囊中进行眼表麻醉。将动物放在立体显微镜下的手术台上，玻璃体注射5uL/眼生理盐水或供试品化合物(化合物1、6、22、73)，30min后，玻璃体内第二次注射5uL/眼生理盐水或CA-I(100ng/眼)造模(单眼)。肌注盐酸苯嗯唑(Idzoxan Hydrochloride)注射液使动物正常醒来。然后将它们放回笼中，并每天3次给予抗生素眼药水。The animals were anesthetized by intramuscular injection of Zoletil and Xylazine at doses of 5-15 mg/kg and 2-12 mg/kg, and Oxybuprocaine eye drops were instilled into the conjunctival sac. Table anesthesia. Place the animal on the operating table under a stereo microscope, and inject 5uL/eye normal saline or test compound (compounds 1, 6, 22, 73) into the vitreous body. After 30 minutes, inject the second time 5uL/eye normal saline or CA-I (100ng/eye) model (monocular). Intramuscular injection of Idzoxan Hydrochloride injection made the animals wake up normally. They were then returned to their cages and antibiotic eye drops were given 3 times a day.
5、光学相干断层扫描(OCT)检查5. Optical coherence tomography (OCT) inspection
检查方法：对所有动物双眼在注射前进行一次OCT(Blue Spectrails OCT，海德堡)检查，在建模后约48小时再进行一次OCT检查，记录两次OCT检测的视网膜厚度(μM)。Inspection method: Perform an OCT (Blue Spectrails OCT, Heidelberg) inspection on the eyes of all animals before injection, and then perform an OCT inspection about 48 hours after modeling, and record the retinal thickness (μM) of the two OCT inspections.
6、数据分析6. Data analysis
使用Microsoft Excel软件进行组均值和标准差的计算。每当有两个以上的组时，使用Levene检验以0.05的显着性水平评估组间方差的齐次性。如果发现组方差之间的差异不显着 (p>0.05)，则进行参数单向方差分析(ANOVA)。当通过ANOVA检验表明平均值之间存在显着差异(p≤0.05)时，将Dunnett检验用于进行对照组和每个供试品组之间的组均值比较。结果如图2所示。Use Microsoft Excel software to calculate the group mean and standard deviation. Whenever there are more than two groups, the Levene test is used to evaluate the homogeneity of variance between groups at a significance level of 0.05. If it is found that the difference between group variances is not significant (p>0.05), then a parametric one-way analysis of variance (ANOVA) is performed. When the ANOVA test shows that there is a significant difference between the means (p≤0.05), Dunnett's test is used to compare the group means between the control group and each test product group. The result is shown in Figure 2.
实施例80 视网膜色素上皮细胞ARPE19毒性测试Example 80 ARPE19 Toxicity Test of Retinal Pigment Epithelial Cells
1、细胞系1. Cell lines
视网膜色素上皮细胞ARPE19Retinal Pigment Epithelial Cell ARPE19
2、供试品制备2. Preparation of test product
称取实施例化合物适量，加入DMSO，充分混匀。然后以2％培养基(DMEM-F12，购自Gibco)进行稀释，获得终浓度分别为1μM和10μM的样品溶液。Weigh an appropriate amount of the compound of the example, add DMSO, and mix well. Then it was diluted with 2% medium (DMEM-F12, purchased from Gibco) to obtain sample solutions with final concentrations of 1 μM and 10 μM, respectively.
3、实验过程：3. Experimental process:
取细胞密度为1*10 5/mL的ARPE19细胞，100μL/孔接种于96孔板，细胞贴壁过夜后，吸除培养板中旧培养基，每孔加入90μL 2％培养基，其中空白组和对照组加入100μL 2％培养基；每孔加入10μL(空白组和对照组除外)供试品溶液，充分混匀，置于培养箱中继续培养24h；24h后，每孔加入10μL CCK-8(C0039，碧云天)溶液(避光)，充分混匀，置于培养箱中继续培养2h；2h后，450nm测OD值。 Take ARPE19 cells with a cell density of 1*10 5 /mL, and inoculate 100 μL/well in a 96-well plate. After the cells adhere to the wall overnight, remove the old medium from the culture plate and add 90 μL of 2% medium to each well. The blank group Add 100μL of 2% medium to the control group; add 10μL of test solution to each well (except the blank group and the control group), mix well, and place in the incubator to continue culturing for 24h; after 24h, add 10μL of CCK-8 to each well (C0039, Biyuntian) solution (protected from light), mix well, place in an incubator and continue to incubate for 2h; after 2h, measure the OD value at 450nm.
4、实验结果4. Experimental results
实验结果如图3所示，结果表明1μM和10μM的实施例化合物均对视网膜色素上皮细胞没有毒性。The experimental results are shown in Fig. 3, and the results show that neither 1 μM nor 10 μM of the compounds of the examples are toxic to retinal pigment epithelial cells.

Claims (29)

  1. 一种式(I)所示的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药：A compound represented by formula (I), its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug:
    Figure PCTCN2020137640-appb-100001
    Figure PCTCN2020137640-appb-100001
    式中，Where
    A选自5元或6元芳环或芳杂环，所述芳杂环含有1-3个选自N、O和S的杂原子，所述芳环或芳杂环任选地被下述取代基取代：卤素、烷基、烷氧基、卤代烷基、OH、CN、COOR 1、CONR 1R 2、NR 1R 2、NR 1COR 2、(CH 2) 1-3NR 1R 2、(CH 2) 1-3OR 1和C(R 1)(R 2)NH 2； A is selected from a 5-membered or 6-membered aromatic ring or an aromatic heterocyclic ring, the aromatic heterocyclic ring containing 1-3 heteroatoms selected from N, O and S, and the aromatic ring or aromatic heterocyclic ring is optionally Substituent substitution: halogen, alkyl, alkoxy, haloalkyl, OH, CN, COOR 1 , CONR 1 R 2 , NR 1 R 2 , NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1-3 OR 1 and C(R 1 )(R 2 )NH 2 ;
    或者A选自稠合的6，5-或6，6-芳杂双环，所述芳杂双环含有N和任选的另外独立地1-2个选自N、O和S的杂原子，所述芳杂双环任选地被下述取代基取代：所述芳环或芳杂环任选地被下述取代基取代：卤素、烷基、烷氧基、卤代烷基、OH、CN、COOR 1、CONR 1R 2、NR 1R 2、NR 1COR 2、(CH 2) 1-3NR 1R 2、(CH 2) 1-3OR 1和-C(R 1)(R 2)NH 2； Or A is selected from a fused 6,5- or 6,6-aromatic heterobicyclic ring containing N and optionally additionally independently 1-2 heteroatoms selected from N, O and S, so The aromatic heterobicyclic ring is optionally substituted with the following substituents: the aromatic ring or aromatic heterocyclic ring is optionally substituted with the following substituents: halogen, alkyl, alkoxy, haloalkyl, OH, CN, COOR 1 , CONR 1 R 2 , NR 1 R 2 , NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1-3 OR 1 and -C(R 1 )(R 2 )NH 2 ；
    L 1选自(CR 3R 4) m或者(CR 3R 4) mO，其中m为0、1、2； L 1 is selected from (CR 3 R 4 ) m or (CR 3 R 4 ) m O, where m is 0, 1, 2;
    X 1和X 2为CR 5或N，且不同时为N；其中R 5选自H、OH、卤素、烷基、烷氧基、卤代烷基、环烷基、CN、COOR 1、CONR 1R 2、NR 1R 2、NR 1COR 2、(CH 2) 1-3NR 1R 2、(CH 2) 1-3OR 1和-C(R 1)(R 2)NH 2； X 1 and X 2 are CR 5 or N, and are not N at the same time; wherein R 5 is selected from H, OH, halogen, alkyl, alkoxy, haloalkyl, cycloalkyl, CN, COOR 1 , CONR 1 R 2. NR 1 R 2 , NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1-3 OR 1 and -C(R 1 )(R 2 )NH 2 ;
    -C-D-选自-NH-CH 2-、-N＝CH-、-NCH 3-CH 2-、-NHCO-、-CH＝CH-或-CH 2-CH 2-； -CD- is selected from -NH-CH 2 -, -N=CH-, -NCH 3 -CH 2 -, -NHCO-, -CH=CH- or -CH 2 -CH 2 -;
    L 2选自键、(CH 2) n、(CH 2) nNH、(CH 2) nO，其中n为1或2； L 2 is selected from bond, (CH 2 ) n , (CH 2 ) n NH, (CH 2 ) n O, where n is 1 or 2;
    B选自5-6元芳环或者芳杂环，其中所述芳杂环含有1-3选自N、O或S的杂原子，任选地，所述芳环或者芳杂环被卤素、烷基、烷氧基、卤代烷基、OH、CN、酮基、COOR 1、CONR 1R 2、NR 1R 2、NR 1COR 2、(CH 2) 1-3NR 1R 2、(CH 2) 1-3OR 1和-C(R 1)(R 2)NH 2取代； B is selected from a 5-6 membered aromatic ring or an aromatic heterocyclic ring, wherein the aromatic heterocyclic ring contains 1-3 heteroatoms selected from N, O or S. Optionally, the aromatic ring or the aromatic heterocyclic ring is halogenated, Alkyl, alkoxy, haloalkyl, OH, CN, keto, COOR 1 , CONR 1 R 2 , NR 1 R 2 , NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1-3 OR 1 and -C(R 1 )(R 2 )NH 2 substitution;
    或者B选自饱和的或不饱和的5-10元单环或稠合多环非芳族环系，所述非芳族环系任选地含有0-3个选自N、O和S的杂原子，所述非芳族环系任选地被下述取代基取代：卤素、烷基、烷氧基、卤代烷基、酮基、OH、CN、COOR 1、CONR 1R 2、NR 1R 2、NR 1COR 2、(CH 2) 1-3NR 1R 2、(CH 2) 1-3OR 1和-C(R 1)(R 2)NH 2； Or B is selected from a saturated or unsaturated 5-10 membered monocyclic or fused polycyclic non-aromatic ring system, the non-aromatic ring system optionally contains 0-3 selected from N, O and S Heteroatoms, the non-aromatic ring system is optionally substituted by the following substituents: halogen, alkyl, alkoxy, haloalkyl, keto, OH, CN, COOR 1 , CONR 1 R 2 , NR 1 R 2. NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1-3 OR 1 and -C(R 1 )(R 2 )NH 2 ;
    R 1和R 2独立地选自H和烷基，或者R 1和R 2与它们所连接的碳一起形成环烷基； R 1 and R 2 are independently selected from H and alkyl, or R 1 and R 2 together with the carbon to which they are attached form a cycloalkyl;
    R 3和R 4独立地选自H和烷基，或者R 3和R 4与它们所连接的碳一起形成环烷基。 R 3 and R 4 are independently selected from H and alkyl, or R 3 and R 4 together with the carbon to which they are attached form a cycloalkyl.
  2. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，A选自The compound, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug according to claim 1, wherein A is selected from
    Figure PCTCN2020137640-appb-100002
    其中，X 3为CR 10或N，R 6、R 7、R 8、R 9、R 10独立地选自氢、卤素、烷基、烷氧基、卤代烷基、OH、CN、COOR 1、CONR 1R 2、NR 1R 2、NR 1COR 2、(CH 2) 1-3NR 1R 2、(CH 2) 1-3OR 1和C(R 1)(R 2)NH 2；R 1和R 2独立地选自H和烷基，或者R 1和R 2与它们所连接的碳一起形成环烷基。
    Figure PCTCN2020137640-appb-100002
    Wherein, X 3 is CR 10 or N, R 6 , R 7 , R 8 , R 9 , R 10 are independently selected from hydrogen, halogen, alkyl, alkoxy, haloalkyl, OH, CN, COOR 1 , CONR 1 R 2 , NR 1 R 2 , NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1-3 OR 1 and C(R 1 )(R 2 )NH 2 ; R 1 And R 2 are independently selected from H and an alkyl group, or R 1 and R 2 together with the carbon to which they are attached form a cycloalkyl group.
  3. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，R 8为NH 2或者C(R 1)(R 2)NH 2，其中R 1和R 2独立地选自H和C 1-3的烷基，或者R 1和R 2与它们所连接的碳一起形成3元或4元环烷基。 The compound, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug according to claim 1, wherein R 8 is NH 2 or C(R 1 )(R 2 )NH 2 , Wherein R 1 and R 2 are independently selected from H and C 1-3 alkyl groups, or R 1 and R 2 together with the carbon to which they are attached form a 3-membered or 4-membered cycloalkyl group.
  4. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，R 8为NH 2。 The compound, or a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof according to claim 1, wherein R 8 is NH 2 .
  5. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，R 8为CH 2NH 2。 The compound, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug according to claim 1, wherein R 8 is CH 2 NH 2 .
  6. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，R 8为C(R 1)(R 2)NH 2，其中R 1和R 2与它们所连接的碳一起形成环丙基。 The compound, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug according to claim 1, wherein R 8 is C(R 1 )(R 2 )NH 2 , wherein R 1 And R 2 together with the carbon to which they are attached form a cyclopropyl group.
  7. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，R 6、R 7、R 9、R 10独立地选自H、卤素、烷基、烷氧基、卤代烷基。 The compound, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug according to claim 1, wherein R 6 , R 7 , R 9 , R 10 are independently selected from H, halogen , Alkyl, alkoxy, haloalkyl.
  8. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，R 6、R 7、R 9、R 10独立地选自H、卤素或者CH 3。 The compound, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug according to claim 1, wherein R 6 , R 7 , R 9 , R 10 are independently selected from H, halogen Or CH 3 .
  9. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，A选自：The compound, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug according to claim 1, wherein A is selected from:
    Figure PCTCN2020137640-appb-100003
    Figure PCTCN2020137640-appb-100003
  10. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，A选自异喹啉和1H-吡咯[2,3-b]吡啶，其中异喹啉和1H-吡咯[2,3-b]吡啶任选地被卤素、烷基、烷氧基、卤代烷基、OH、CN、COOR 1、CONR 1R 2、NR 1R 2、NR 1COR 2、(CH 2) 1-3NR 1R 2、(CH 2) 1-3OR 1和C(R 1)(R 2)NH 2取代；R 1和R 2独立地选自H和烷基，或者R 1和R 2与它们所连接的碳一起形成环烷基。 The compound, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug according to claim 1, wherein A is selected from isoquinoline and 1H-pyrrole [2,3-b]pyridine , Wherein isoquinoline and 1H-pyrrole [2,3-b]pyridine are optionally substituted by halogen, alkyl, alkoxy, haloalkyl, OH, CN, COOR 1 , CONR 1 R 2 , NR 1 R 2 , NR 1 COR 2 , (CH 2 ) 1-3 NR 1 R 2 , (CH 2 ) 1-3 OR 1 and C(R 1 )(R 2 )NH 2 are substituted; R 1 and R 2 are independently selected from H And alkyl, or R 1 and R 2 together with the carbon to which they are attached form a cycloalkyl group.
  11. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，A选自异喹啉和1H-吡咯[2,3-b]吡啶，其中异喹啉和1H-吡咯[2,3-b]吡啶任选地被卤素、烷基、烷氧基、卤代烷基和氨基取代。The compound, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug according to claim 1, wherein A is selected from isoquinoline and 1H-pyrrole [2,3-b]pyridine , Wherein isoquinoline and 1H-pyrrole [2,3-b]pyridine are optionally substituted by halogen, alkyl, alkoxy, haloalkyl and amino.
  12. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，A选自：The compound, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug according to claim 1, wherein A is selected from:
    Figure PCTCN2020137640-appb-100004
    Figure PCTCN2020137640-appb-100004
  13. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，L 1选自键、CH 2、(CH 2) 2O。 The compound, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug according to claim 1, wherein L 1 is selected from bond, CH 2 , (CH 2 ) 2 O.
  14. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，L 1选自CH 2。 The compound, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug according to claim 1, wherein L 1 is selected from CH 2 .
  15. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，X 2为CR 5，其中R 5选自H、烷基、卤代烷基和烷氧基。 The compound according to claim 1, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug, wherein X 2 is CR 5 , wherein R 5 is selected from H, alkyl, haloalkyl And alkoxy.
  16. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，X 2为CR 5，其中R 5选自H、CH 3、CF 3、CH 2OCH 3。 The compound, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug of claim 1, wherein X 2 is CR 5 , wherein R 5 is selected from H, CH 3 , CF 3 , CH 2 OCH 3 .
  17. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，-C-D-选自-NH-CH 2-、-N＝CH-、-NHCO-。 The compound, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug according to claim 1, wherein -CD- is selected from -NH-CH 2 -, -N=CH-, -NHCO-.
  18. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，L 2选自键、O、NH、CH 2、CH 2NH、CH 2O。 The compound of claim 1, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug, wherein L 2 is selected from bond, O, NH, CH 2 , CH 2 NH, CH 2 O.
  19. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，L 2选自CH 2。 The compound, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug according to claim 1, wherein L 2 is selected from CH 2 .
  20. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，B选自：The compound, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug according to claim 1, wherein B is selected from:
    Figure PCTCN2020137640-appb-100005
    Figure PCTCN2020137640-appb-100005
  21. 如权利要求1所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于所述化合物选自：The compound, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug according to claim 1, characterized in that the compound is selected from:
    Figure PCTCN2020137640-appb-100006
    Figure PCTCN2020137640-appb-100006
    Figure PCTCN2020137640-appb-100007
    Figure PCTCN2020137640-appb-100007
  22. 一种药物组合物，其特征在于包含权利要求1-21中任一项所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药，和药学上可接受的赋形剂。A pharmaceutical composition characterized by comprising the compound according to any one of claims 1-21, its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug, and pharmaceutically acceptable excipients. Shape agent.
  23. 如权利要求1-21中任一项所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药在制备药物中的用途，所述药物用于预防或治疗涉及血浆激肽释放酶活性相关的疾病。Use of the compound, pharmaceutically acceptable salt, solvate, stereoisomer or prodrug of any one of claims 1-21 in the preparation of a medicament for prevention or treatment involving plasma Diseases related to kallikrein activity.
  24. 如权利要求1-21中任一项所述的化合物、其药学上可接受的盐、溶剂化物、立体异构体或前药用于预防或者治疗涉及血浆激肽释放酶活性相关的疾病的方法，包括向有需要的哺乳动物施用治疗有效量的权利要求1-21中任一项所述的化合物。The compound according to any one of claims 1-21, a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof for use in a method for preventing or treating diseases related to plasma kallikrein activity , Comprising administering a therapeutically effective amount of the compound of any one of claims 1-21 to a mammal in need.
  25. 根据权利要求23所述的用途或者权利要求24所述的预防或者治疗方法，其特征在于所述涉及血浆激肽释放酶活性的疾病为炎症。The use according to claim 23 or the prevention or treatment method according to claim 24, characterized in that the disease involving plasma kallikrein activity is inflammation.
  26. 根据权利要求23所述的用途或者权利要求24所述的预防或者治疗方法，其特征在于所述涉及血浆激肽释放酶活性的疾病选自视力受损、糖尿病视网膜病变、糖尿病黄斑水肿、遗传性血管水肿、糖尿病、胰腺炎、脑出血、肾病、心肌病、神经病、炎性肠病、关节炎、感染性休克、低血压、癌症、成人呼吸窘迫综合征、弥散性血管内凝血、心肺旁路手术和外科手术后出血。The use according to claim 23 or the prevention or treatment method according to claim 24, wherein the disease involving plasma kallikrein activity is selected from the group consisting of impaired vision, diabetic retinopathy, diabetic macular edema, hereditary Angioedema, diabetes, pancreatitis, cerebral hemorrhage, nephropathy, cardiomyopathy, neuropathy, inflammatory bowel disease, arthritis, septic shock, hypotension, cancer, adult respiratory distress syndrome, diffuse intravascular coagulation, cardiopulmonary bypass Bleeding after surgery and surgery.
  27. 根据权利要求23所述的用途或者权利要求24所述的预防或者治疗方法，其特征在于所述涉及血浆激肽释放酶活性的疾病是与糖尿病视网膜病变和糖尿病黄斑水肿相关的视网膜血管通透性疾病。The use according to claim 23 or the prevention or treatment method according to claim 24, wherein the disease involving plasma kallikrein activity is retinal vascular permeability related to diabetic retinopathy and diabetic macular edema disease.
  28. 如权利要求23所述的用途或者权利要求24所述的预防或者治疗方法，其特征在于所述涉及血浆激肽释放酶活性的疾病为糖尿病黄斑水肿。The use according to claim 23 or the prevention or treatment method according to claim 24, wherein the disease involving plasma kallikrein activity is diabetic macular edema.
  29. 如权利要求23所述的用途或者权利要求24所述的预防或者治疗方法，其特征在于所述涉及血浆激肽释放酶活性的疾病为遗传性血管水肿。The use according to claim 23 or the prevention or treatment method according to claim 24, wherein the disease involving plasma kallikrein activity is hereditary angioedema.
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