WO2018228452A1 - 一种抑制 rock 的化合物及其用途 - Google Patents

一种抑制 rock 的化合物及其用途 Download PDF

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WO2018228452A1
WO2018228452A1 PCT/CN2018/091161 CN2018091161W WO2018228452A1 WO 2018228452 A1 WO2018228452 A1 WO 2018228452A1 CN 2018091161 W CN2018091161 W CN 2018091161W WO 2018228452 A1 WO2018228452 A1 WO 2018228452A1
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ring
membered
group
membered aromatic
mmol
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PCT/CN2018/091161
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French (fr)
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李进
张登友
冯静超
廖伟
林丽
李偲
陈伟
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成都先导药物开发有限公司
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Priority to EP18818595.3A priority Critical patent/EP3640242B1/en
Priority to JP2020519170A priority patent/JP6906105B2/ja
Publication of WO2018228452A1 publication Critical patent/WO2018228452A1/zh
Priority to US16/716,455 priority patent/US11078161B2/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/472—Non-condensed isoquinolines, e.g. papaverine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/472—Non-condensed isoquinolines, e.g. papaverine
    • A61K31/4725—Non-condensed isoquinolines, e.g. papaverine containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A61P27/06—Antiglaucoma agents or miotics
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D217/00—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems
    • C07D217/02—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems with only hydrogen atoms or radicals containing only carbon and hydrogen atoms, directly attached to carbon atoms of the nitrogen-containing ring; Alkylene-bis-isoquinolines
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
    • C07D409/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • the invention belongs to the field of drug synthesis, and in particular relates to a compound for inhibiting ROCK and its application in treating diseases related to ROCK.
  • Rho belongs to the small molecule monomeric GTPase superfamily and is a mammalian gene homolog of the Ras superfamily. It regulates cell muscle through its downstream major Rho-associated coiled-coil containing protein kinase (ROCK).
  • ROCK protein kinase
  • the reorganization of the kinesin backbone is widely involved in a series of biological processes such as cell mitosis, cytoskeletal regulation, smooth muscle cell contraction, nerve regeneration, tumor cell infiltration, and apoptosis regulation.
  • Rho/ROCK When Rho/ROCK is activated, it can act on a variety of substrates to produce biological processes.
  • MLC myosin light chain
  • MLCP myosin light chain phosphatase
  • Rho kinase activity in animal models exhibits a variety of benefits in the treatment of human diseases, including cardiovascular diseases such as pulmonary hypertension, hypertension, atherosclerosis, cardiac hypertrophy, ocular hypertension, cerebral ischemia, cerebral vasospasm, etc. , and central nervous system disorders such as neuronal degeneration.
  • cardiovascular diseases such as pulmonary hypertension, hypertension, atherosclerosis, cardiac hypertrophy, ocular hypertension, cerebral ischemia, cerebral vasospasm, etc.
  • central nervous system disorders such as neuronal degeneration.
  • ROCK expression and activity are elevated in spontaneously hypertensive rats, indicating that it is associated with the development of hypertension in these animals (Involvement of Rho-kinase in hypertensive vascular disease: a novel therapeutic target in hypertension [J] .FASEB J., 2001, 15(6): 1062-4).
  • the ROCK inhibitor Y-27632 can significantly reduce blood pressure in three rat hypertension models (spontaneous hypertension, renal hypertension, deoxycorticosterone acetate hypertension), while the blood pressure of control rats is higher.
  • ROCK inhibitors have a good effect on pulmonary hypertension (Acute vasodilator effects of a Rho-kinase inhibitor, fasudil, in patients with severe pulmonary hypertension [J]. Heart, 2005: 91 (3): 391-2) .
  • Isoquinolines These compounds are structurally characterized by having an isoquinoline structure and a piperazine ring, both of which are linked by a sulfonyl group. Representatives are Faithil (Uehata M, Ishizaki T, Satoh H, et al. Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension [J]. Nature, 1997, 389: 990-994), H-1152P (Tamura M, Nakao H, Yoshizaki H, et al. Development of specific Rho-kinase inhibitors and their clinical application [J].
  • ROCK inhibitor drugs are Elisa (for the treatment of cerebral vasospasm) from Asahi Kasei and Kowa. (K-115) (for the treatment of ocular hypertension and glaucoma). among them It is only available for sale in Japan. Therefore, it is important to develop a targeted small molecule drug study for ROCK to obtain a ROCK inhibitor with better activity, higher selectivity, longer duration of action, higher stability, lower toxicity and side effects, and more economical. Social and economic significance.
  • the present invention provides a compound of the formula (I) or a stereoisomer thereof:
  • n 0, 1, or 2, respectively;
  • R 1 is each independently selected from the group consisting of hydrogen, hydroxy, halogen, amino, carboxy, trifluoromethyl, nitro, cyano, C 1-6 alkyl;
  • n 0, 1, 2, 3, 4 or 5 respectively;
  • R 3 is selected from -NR 2 R 2 ' , N-containing heterocycloalkyl; wherein the N-containing heterocycloalkyl group may be further substituted by 1 to 2 substituents independently selected from halogen, C 1-6 alkyl Replace
  • R 2 and R 2′ are each independently selected from hydrogen and C 1-6 alkyl
  • Ring A is selected from a 5- to 6-membered aromatic ring, a 5- to 6-membered aromatic heterocyclic ring or
  • the B ring and the C ring are each independently selected from a 5- to 6-membered aromatic ring and a 5- to 6-membered aromatic heterocyclic ring;
  • R 4 is each independently selected from the group consisting of hydrogen, halogen, nitro, cyano, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, 3- to 6-membered cycloalkyl, 3 to 6 Heterocycloalkyl, -(CH 2 ) m OR a , -(CH 2 ) m OC(O)R a , -(CH 2 ) m OC(O)NR a R b , -(CH 2 ) m NR a R b , -(CH 2 ) m NR a C(O)R b , -(CH 2 ) m NR a C(O)OR b , -(CH 2 ) m C(O)R a , -(CH 2 m C(O)OR a , -(CH 2 ) m C(O)NR a R b ;
  • R a and R b are each independently selected from the group consisting of hydrogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, 3- to 6-membered cycloalkyl, 3- to 6-membered heterocycloalkyl, and 5 ⁇ 6-membered aromatic ring and 5- to 6-membered aromatic heterocyclic ring; wherein, 3 to 6-membered cycloalkyl group, 3 to 6-membered heterocycloalkyl group, 5- to 6-membered aromatic ring, and 5- to 6-membered aromatic heterocyclic ring can be further 1 to 2 substituents independently selected from halogen, C 1 ⁇ 6 alkyl substituents.
  • the N-containing heterocycloalkyl group selected from R 3 is a 4- to 5-membered N-containing heterocycloalkyl group
  • R 3 is selected from the group consisting of N-containing heterocycloalkyl groups and the A ring has 2 R 4 substitutions, 2 R 4 are not simultaneously halogen;
  • the present invention also provides a compound of the formula (II) or a stereoisomer thereof:
  • R 1 is selected from the group consisting of hydrogen, hydroxy, halogen, amino, carboxy, trifluoromethyl, nitro, cyano, C 1-6 alkyl;
  • n 0, 1, 2 or 3 respectively;
  • R 2 and R 2′ are each independently selected from hydrogen and C 1-6 alkyl
  • Ring A is selected from a 5- to 6-membered aromatic ring, a 5- to 6-membered aromatic heterocyclic ring or
  • the B ring and the C ring are each independently selected from a 5- to 6-membered aromatic ring and a 5- to 6-membered aromatic heterocyclic ring;
  • R 4 is each independently selected from the group consisting of hydrogen, halogen, nitro, cyano, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, 3- to 6-membered cycloalkyl, 3 to 6 Heterocycloalkyl, -(CH 2 ) m OR a , -(CH 2 ) m OC(O)R a , -(CH 2 ) m NR a R b , -(CH 2 ) m NR a C(O) R b , -(CH 2 ) m C(O)R a , -(CH 2 ) m C(O)OR a , -(CH 2 ) m C(O)NR a R b ;
  • R a and R b are each independently selected from the group consisting of hydrogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, 3- to 6-membered cycloalkyl, 3- to 6-membered heterocycloalkyl, and 5 ⁇ 6-membered aromatic ring and 5- to 6-membered aromatic heterocyclic ring; wherein, 3 to 6-membered cycloalkyl group, 3 to 6-membered heterocycloalkyl group, 5- to 6-membered aromatic ring, and 5- to 6-membered aromatic heterocyclic ring can be further 1 to 2 substituents independently selected from halogen, C 1 ⁇ 6 alkyl substituents.
  • the compound of formula (II) is as shown in the following structural formula:
  • the invention also provides a compound of the formula (III) or a stereoisomer thereof:
  • R 1 is selected from the group consisting of hydrogen, hydroxy, halogen, amino, carboxy, trifluoromethyl, nitro, cyano, C 1-6 alkyl;
  • n 0, 1, 2 or 3 respectively;
  • Ring A is selected from a 5- to 6-membered aromatic ring, a 5- to 6-membered aromatic heterocyclic ring or
  • the B ring and the C ring are each independently selected from a 5- to 6-membered aromatic ring and a 5- to 6-membered aromatic heterocyclic ring;
  • the ring E is selected from a heterocycloalkyl group containing N; the N-containing heterocycloalkyl group may be further substituted by 1 to 2 substituents independently selected from halogen, C 1-6 alkyl;
  • R 4 is each independently selected from the group consisting of hydrogen, halogen, nitro, cyano, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, 3- to 6-membered cycloalkyl, 3 to 6 Heterocycloalkyl, -(CH 2 ) m OR a , -(CH 2 ) m OC(O)R a , -(CH 2 ) m NR a R b , -(CH 2 ) m NR a C(O) R b , -(CH 2 ) m C(O)R a , -(CH 2 ) m C(O)OR a , -(CH 2 ) m C(O)NR a R b ;
  • R a and R b are each independently selected from the group consisting of hydrogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, 3- to 6-membered cycloalkyl, 3- to 6-membered heterocycloalkyl, and 5 ⁇ 6-membered aromatic ring and 5- to 6-membered aromatic heterocyclic ring; wherein, 3 to 6-membered cycloalkyl group, 3 to 6-membered heterocycloalkyl group, 5- to 6-membered aromatic ring, and 5- to 6-membered aromatic heterocyclic ring can be further 1 to 2 substituents independently selected from halogen, C 1 ⁇ 6 alkyl substituents.
  • the E ring is selected from the group consisting of 4 to 5 membered N-containing heterocycloalkyl groups.
  • the disease associated with abnormal ROCK activity is a disease associated with cytoskeletal regulation, smooth muscle cell contraction, and nerve regeneration.
  • the disease is ocular hypertension or glaucoma.
  • a medicament comprising the above compound or a stereoisomer, a crystal form, a pharmaceutically acceptable salt, a hydrate or a solvate thereof as an active ingredient, plus a pharmaceutically acceptable substance A preparation prepared from the accepted excipients.
  • the compounds and derivatives provided in the present invention may be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature system.
  • substitution means that a hydrogen atom in a molecule is replaced by a different atom or molecule.
  • the minimum and maximum values of the carbon atom content in the hydrocarbon group are represented by a prefix, for example, the prefix C a to b alkyl group indicates any alkyl group having "a" to "b" carbon atoms.
  • C 1 - 4 alkyl means an alkyl group containing from 1 to 4 carbon atoms.
  • the alkenyl group in the present invention may contain one or more carbon-carbon double bonds.
  • the alkynyl group in the present invention may contain one or more carbon-carbon triple bonds.
  • the C a to b alkoxy group, the C a to b alkyl ester group, the C a to b alkylamino group, and the C a to b acyl group respectively mean an alkyl group having "a" to "b" carbon atoms and corresponding a group obtained by linking an oxygen atom, an ester group, an amino group, or an acyl group.
  • Stereoisomers in the present invention include enantiomers and diastereomers.
  • the halogen in the present invention includes a fluorine atom, a chlorine atom, a bromine atom, an iodine atom and the like.
  • the hetero atom in the present invention includes a nitrogen atom, an oxygen atom, a sulfur atom and the like.
  • the cycloalkyl group in the present invention means a non-aromatic cyclic hydrocarbon group including a saturated cycloalkyl group and a partially saturated cycloalkyl group.
  • the heterocycloalkyl group of the present invention has a hetero atom-containing non-aromatic cycloalkyl group, and includes a saturated heterocycloalkyl group and a partially saturated heterocycloalkyl group.
  • the aromatic ring in the present invention means an aromatic hydrocarbon group having an aromatic character.
  • the aromatic heterocyclic ring in the present invention means an aromatic ring containing a hetero atom in the ring.
  • pharmaceutically acceptable means that a carrier, carrier, diluent, adjuvant, and/or salt formed is generally chemically or physically compatible with the other ingredients that constitute a pharmaceutical dosage form, and is physiologically Compatible with the receptor.
  • salts and “pharmaceutically acceptable salt” refer to the above-mentioned compounds or stereoisomers thereof, acid and/or basic salts formed with inorganic and/or organic acids and bases, and also includes zwitterionic salts (within Salts) also include quaternary ammonium salts such as alkylammonium salts. These salts can be obtained directly in the final isolation and purification of the compounds. It can also be obtained by mixing the above compound, or a stereoisomer thereof, with a certain amount of an acid or a base as appropriate (for example, an equivalent amount).
  • the salt in the present invention may be a hydrochloride, a sulfate, a citrate, a besylate, a hydrobromide, a hydrofluoride, a phosphate, an acetate, a propionate or a dibutyl compound.
  • one or more compounds of the invention may be used in combination with one another.
  • the compounds of the invention may be used in combination with any other active agent for the preparation of a medicament or pharmaceutical composition that modulates cellular function or treats a disease. If a group of compounds is used, these compounds can be administered to the subject simultaneously, separately or sequentially.
  • Figure 1 shows the experimental results of K115 on the ocular hypotensive activity of New Zealand white rabbits with normal intraocular pressure.
  • Figure 2 is the experimental results of the ocular hypotensive activity of compound 5 against normal intraocular pressure New Zealand white rabbits.
  • Figure 3 is the experimental results of the ocular hypotensive activity of compound 11a against normal intraocular pressure New Zealand white rabbits.
  • Figure 4 is the experimental results of ocular hypotensive activity of compound 11a against high intraocular pressure New Zealand white rabbits.
  • the structure of the compound was determined by nuclear magnetic resonance (NMR) and mass spectrometry (MS).
  • the NMR shift ( ⁇ ) is given in units of 10-6 (ppm).
  • NMR was measured using a (Bruker Avance III 400 and Bruker Avance 300) nuclear magnetic apparatus, and the solvent was deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl 3 ), deuterated methanol (MeOD), internal standard. It is tetramethylsilane (TMS).
  • the LC-MS was measured using a Shimadzu LC-MS 2020 (ESI).
  • the HPLC was measured using Shimadzu High Pressure Liquid Chromatograph (Shimadzu LC-20A).
  • MPLC medium pressure preparative chromatography
  • the thin layer chromatography silica gel plate is separated from the Yantai Yellow Sea HSGF254 or Qingdao GF254 silica gel plate by thin layer chromatography, and the specification is 0.4mm to 0.5mm.
  • the raw materials and equipment used in the specific embodiments of the present invention are known products and are obtained by purchasing commercially available products.
  • Known starting materials can be synthesized by or according to methods known in the art, or can be purchased from companies such as Anike Chemical, Chengdu Kelon Chemical, Suiyuan Chemical Technology, and Belling Technology.
  • the room temperature is an optimum reaction temperature of 20 ° C to 30 ° C.
  • M is a mole per liter.
  • the solution means an aqueous solution.
  • DCM dichloromethane
  • PE petroleum ether
  • THF tetrahydrofuran
  • DMF N,N-dimethylformamide
  • DIEA diisopropylethylamine
  • DMAP 4 -dimethylaminopyridine
  • EDCI 1-ethyl-(3-dimethylaminopropyl)carbonyldiimide hydrochloride
  • HOBT 1-hydroxybenzotriazole
  • HBTU benzotriazole- N,N,N',N'-tetramethyluronium hexafluorophosphate
  • Pd(dppf)Cl 2 is [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride.
  • Benzyl-N-((())-tert-butylsulfenamide)-2-phenethyl)carbamate (5.00 g, 13.4 mmol) was dissolved in methanol (10.0 mL). A solution of dioxane in hydrochloric acid (40.0 ml, 5 mol/L) was stirred for 1 hour, and then the solvent was evaporated under reduced pressure to give benzyl-N-(2-amino-2-phenyl-ethyl)carbamate (3.01) g, 10.2 mmol, yield 76%).
  • Benzyl-N-(2-amino-2-phenyl-ethyl)carbamate (2.00 g, 7.40 mmol) was dissolved in DMF (10.0 mL) and benzotriazole-N, N, N was added.
  • ',N'-Tetramethylurea hexafluorophosphate (2.07 g, 8.14 mmol)
  • isoquinoline-6-carboxylic acid (1.28 g, 7.40 mmol
  • N,N-diisopropylethylamine (3.38 g, After stirring for 1 hour, the mixture was extracted with EtOAc EtOAc.
  • 6-Isoquinolinecarboxylic acid (1.50 g, 8.30 mmol) was dissolved in DMF (15.0 mL).
  • N-(1-(3-Chlorophenyl)-2-cyanoethyl)isoquinolin-6-carboxamide 70.0 mg, 210 ⁇ mol was dissolved in methanol (5.00 mL), and Raney nickel (10.0) was added at room temperature. Mg). Stir at room temperature for 3 hours under hydrogen. The solid was removed by filtration, and the solvent was evaporated under reduced pressure. Purification by MPLC gave N-(3-amino-1-(3-chlorophenyl)propyl)isoquinoline-6-carboxamide (15.2 mg, 42.0 [mu]mol, yield 20%).
  • tert-Butyl (1-(3-chlorophenyl)-2-cyanoethyl)carbamate (2.00 g, 7.12 mmol) was dissolved in methanol (25.0 mL), and Raney nickel (200 mg) was added at room temperature. Stir at room temperature for 3 hours under hydrogen. The solid was removed by filtration, and the solvent was evaporated under reduced pressure. Purification by MPLC gave methyl tert-butyl(3-amino-1-(3-chlorophenyl)propyl)carbamate (2.01 g, 7.02 mmol, yield 99%).
  • Benzyl tert-butyl (1-(3-chlorophenyl)propane-1,3-substituted) dicarbamate (2.20 g, 5.25 mmol) was dissolved in 1,4-dioxane (30.0 mL) Adding pinacol borate (5.34 g, 421 mmol), 2-biscyclohexylphosphine-2', 6'-dimethoxybiphenyl (43.1 mg, 110 ⁇ mol), palladium acetate (118 mg, 530 ⁇ mol) at room temperature And potassium acetate (1.54 g, 15.8 mmol). Stir at 100 ° C for 4 hours under nitrogen. The solvent was evaporated under reduced pressure and the mixture was evaporated.
  • Step 7 Preparation of ethyl 4-(3-(3-(benzyloxycarboxamide)-1-(isoquinolin-6-carboxamide)propyl)phenyl)thiophene-2-carboxylate
  • 6-Isoquinolinecarboxylic acid (390 mg, 2.26 mmol) was dissolved in DMF (15.0 mL).
  • EtOAc EtOAc, EtOAc, EtOAc, Ethyl 3-(benzyloxyamide)propyl)phenyl)thiophene-2-carboxylate (900 mg, 2.05 mmol).
  • the solvent was evaporated under reduced pressure and the mixture was evaporated.
  • Purification by column chromatography gave ethyl 4-(3-(3-(benzyloxycarboxamide)-1-(isoquinolin-6-carboxamide)propyl)phenyl)thiophene-2-carboxylate (1.01 g, 1.68 mmol, yield 83%).
  • Step 8 Preparation of ethyl 4-(3-(3-amino-1-(isoquinolin-6-carboxamide)propyl)phenyl)thiophene-2-carboxylate
  • tert-Butyl (1-(3-chlorophenyl)-2-cyanoethyl)carbamate (500 mg, 1.78 mmol) was dissolved in methanol (10.0 mL) and palladium carbon (50.0 mg) was added at room temperature. Stir at room temperature for 3 hours under hydrogen. A solution of formaldehyde (535 mg, 17.8 mmol) was added, and stirred at room temperature under hydrogen for 3 hr. The solid was removed by filtration, and the solvent was evaporated under reduced pressure. Purification by column chromatography gave tert-butyl N-(3-(dimethylamino)-1-phenyl-propyl)amino acid methyl ester (402 mg, 1.44 mmol, yield 81%).
  • 6-Isoquinolinecarboxylic acid 100 mg, 570 ⁇ mol was dissolved in DMF (5.00 mL), and HBTU (219 mg, 570 ⁇ mol), DIEA (224 mg, 1.73 mmol) and N', N'-dimethyl-1 benzene were added at room temperature.
  • Base-propyl-1,3-diamine 100 mg, 570 ⁇ mol. Stir at room temperature for 3 hours. The solvent was evaporated under reduced pressure and the mixture was evaporated. Purification by column chromatography gave N-(3-(dimethylamino)-1-phenyl-propyl)isoquinoline-6-carboxamide (39.6 mg, 120 ⁇ mol, yield 21%).
  • Tetra-tert-butyl-N-((1-benzylazetidin-3-yl)-phenyl-methyl)carbamate (1.20 g, 2.80 mmol) was dissolved in methanol (10 mL). Palladium hydroxide (39 mg, 10%) was stirred under hydrogen for 24 hours. After filtration over celite, the solvent was evaporated evaporated evaporated evaporated. mjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj %).
  • Tetra-tert-butyl-N-(azetidin-3-yl(phenyl)methyl)carbamate (1.00 g, 3.81 mmol) was dissolved in tetrahydrofuran (10 mL) and triethylamine (770 mg, 7.62 mmol) and allyl chloroformate (367 g, 3.05 mmol), EtOAc (EtOAc)EtOAc. The solvent was distilled off under reduced pressure. Purification by column chromatography gave allyl-3-((tert-butoxycarbamoyl)-phenyl-methyl)azetidin-1-carboxylate (597 mg, 1.39 mmol, yield 55%).
  • Tetra-tert-butyl-N-(azetidin-3-yl(phenyl)methyl)carbamate (1.00 g, 3.81 mmol) was dissolved in methanol (10.0 mL) and acetic acid (266 mg, 3.81). Methyl acetate and aqueous formaldehyde (572 mg, 19.1 mmol) were stirred for 1 hour then sodium cyanoborohydride (239 mg, 3.81 mmol). After quenching with aq. aq. EtOAc, EtOAc. Purification by column chromatography gave tetra-tert-butyl(methyl-N-azetidin-3-yl(phenyl)methyl)carbamate (796 mg, 2.03 mmol, yield 53%).
  • Tetra-tert-butyl (methyl-N-azetidin-3-yl(phenyl)methyl)carbamate (773 mg, 2.80 mmol) was dissolved in methanol (10 mL). (3.00 ml), after stirring for 2 hours, the solvent was evaporated under reduced pressure to give the allyl-3-(benzylamine) azetidine-1-carboxylate (0.48 g, 2.2 mmol, yield 78%) .
  • tert-Butyl (R)-3-(methoxy(methyl)carbamoyl)tetrahydropyrrole-1-carboxylate 10 g, 38.7 mmol was dissolved in THF (30 mL), phenyl bromide ( 194 mmol, 1 M, 194 mL) was added dropwise to the reaction mixture. The mixture was stirred at 0 ° C for 1 hour, then extracted with ethyl acetate and water, and the aqueous phase was extracted twice with ethyl acetate. Remove solvent. Purification by column chromatography gave tert-butyl(R)-3-benzoyltetrahydropyrrole-l-carboxylate (9.1 g, 33 mmol, 84% yield).
  • Isoquinoline-6-carboxylic acid (3.00 g, 17.3 mmol) was dissolved in DMF (20.0 mL), and benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (7.26) was added. g, 19.06 mmol), tert-butyl(R)-3-(amino(phenyl)methyl)tetrahydropyrrole-1-carboxylate (4.79 g, 17.3 mmol) and N,N-diisopropyl Amine (3.38 g, 29.6 mmol), EtOAc (EtOAc)EtOAc.
  • EtOAc EtOAc
  • N-((R)-Phenyl-((R)-tetrahydropyrrol-3-yl)methyl)isoquinolin-6-carboxamide 250 mg, 1.51 mmol was dissolved in methanol (15.0 mL) Acetic acid (9.1 mg, 0.15 mmol) and aqueous formaldehyde (0.58 mL, 13M, 7.55 mmol). After stirring for one hour, sodium borohydride (0.96 g, 4.53 mmol) was added, and the reaction was continued for 2 hours, then extracted with ethyl acetate and water. The aqueous phase was extracted twice with ethyl acetate.
  • N-((S)-Phenyl-((R)-tetrahydropyrrol-3-yl)methyl)isoquinolin-6-carboxamide 250 mg, 1.51 mmol was dissolved in methanol (15.0 mL) Acetic acid (9.1 mg, 0.15 mmol) and aqueous formaldehyde (0.58 mL, 13M, 7.55 mmol). After stirring for one hour, sodium borohydride (0.96 g, 4.53 mmol) was added, and the reaction was continued for 2 hours, then extracted with ethyl acetate and water. The aqueous phase was extracted twice with ethyl acetate.
  • ROCK2 is capable of phosphorylating S6K (KRRRLASLR) polypeptide substrates and converting ATP to ADP.
  • ADP-Glo TM Reagent kinase reaction was terminated, and consumed the remaining ATP.
  • Kinase assay reagents were added, so that it converts ADP into ATP at the same time, ATP is then converted Ultra-Glo TM luciferase light emission signal to the light emitting signal positively correlated with kinase activity.
  • the ROCK2 inhibitory activity was tested as follows:
  • Detection buffer 40 mM Tris pH 7.5, 20 mM MgCl 2 , 0.1% BSA (w / v), 50 ⁇ M DTT;
  • Example compound ROCK2 Example compound ROCK2 1 +++ 2 +++ 3 ++ 4 ++ 5 +++ 6 +++ 7 +++ 8 +++ 9 +++ 10 +++ 11a +++ 11b +++ 12 +++ 13 +++ 14a ++ 14b +++ 15 +++
  • the rat smooth muscle cell line A7r5 was used. ROCK2 causes changes in the cytoskeleton by phosphorylating two amino acid sites of myosin light chain T18/S19. A7r5 cells were seeded into 96-well clear bottom black plates under the conditions of DMEM containing 10% FBS. After overnight incubation, the serum was starved for 4 hours and the cells were incubated with the compound for 1 hour in serum-free medium.
  • Myosin light chain phosphorylation levels were detected by in situ immunoblotting using phspho-MLC-T18/S19 specific antibodies and a second detection antibody. Untreated cells served as a positive control, primary antibody unincubated samples served as a negative control, and total intracellular proteins served as internal controls. Using GraphPad Prism 5.01 software using a nonlinear regression curve fit with the slope of determined value 50 EC.
  • the compound prepared in the examples was subjected to the detection of the myosin light chain phosphorylation inhibitory activity of the rat smooth muscle cell line A7r5 according to the above method.
  • the test results are shown in Table 1, wherein the IC 50 of each compound was determined according to the description, in Table 1:
  • Example compound A7r5 Example compound A7r5 1 ++ 2 +++ 4 + 5 ++ 6 +++ 7 ++ 8 + 9 ++ 10 ++ 11a ++ 13 ++ 15 ++ 16 ++
  • the compound of the present invention has a good inhibitory activity against the rat smooth muscle cell line A7r5, and further, the compound of the present invention can inhibit the cell from the ROCK2 by phosphorylating the myosin light chain T18/S19 two amino acid sites.
  • the skeleton changes and has the potential to treat diseases associated with this.
  • Test Example 3 animal experiment (normal intraocular pressure model)
  • test compound was weighed and dissolved in physiological saline to a use concentration.
  • test compound, the control substance K115, or the negative control solvent 50 ⁇ L was applied to the right eye of each animal, and the left eye was added with physiological saline.
  • the intraocular pressure was measured after the first Pre, 1, 2, 4, 6, 8, 10 hours after the administration of the drug. Student-t test was used to analyze data at a single time point. The curve of intraocular pressure over time will be analyzed using the ANOVA test. A P value of less than 0.05 will be considered statistically significant, and the data will be presented as a mean plus or minus standard error.
  • Test Example 4 animal experiment (high intraocular pressure single eye modeling model)
  • the intraocular pressure values of both eyes were measured and recorded as basal intraocular pressure.
  • white rabbits were anesthetized by intravenous injection (50%) of sodium pentobarbital 50 mg/kg.
  • Temporary anterior chamber puncture was established using a tip 30-G needle, and 50 ⁇ L of viscous substance was injected into the anterior chamber of the right eye of the white rabbit to induce high intraocular pressure, and the same volume of physiological saline was administered to the lateral eye.
  • a cotton swab is used to press the anterior chamber puncture to prevent backflow of the aqueous humor.
  • test compound was administered to the right eye of each group of model animals, and administered once every three times at 0, 3, and 6 hours, 50 ⁇ L/eye.
  • the same volume of physiological saline or a specific vehicle will be administered to the test eye at the same time point.
  • Intraocular pressure was measured after 0, 1, 2, 3, 4, 6, and 8 hours. Student-t test was used to analyze data at a single time point. The curve of intraocular pressure over time will be analyzed using the ANOVA test. A P value of less than 0.05 will be considered statistically significant, and the data will be presented as a mean plus or minus standard error.
  • the above test of the present invention shows that the compound of the present invention has a good ROCK inhibitory activity and can be effectively used for the treatment of an abnormal disease with ROCK activity.
  • novel compound of the formula I disclosed in the present invention exhibits a good ROCK inhibitory activity, and provides a new medicinal possibility for clinical treatment of diseases associated with abnormal ROCK activity.

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Abstract

提供了具有ROCK抑制活性的式（I）化合物、该化合物的制备方法以及在制备治疗与ROCK活性异常相关疾病的药物中的用途。

Description

一种抑制ROCK的化合物及其用途 技术领域
本发明属于药物合成领域，具体涉及一种抑制ROCK的化合物及其在治疗与ROCK相关疾病中的应用。
背景技术
Rho属于小分子单聚体GTPase超家族，是Ras超家族的哺乳动物基因同系物，通过其下游最主要的效应分子Rho激酶(Rho-associated coiled-coil containing protein kinase，ROCK)，来调节细胞肌动蛋白骨架的重组，从而广泛参与细胞有丝分裂、细胞骨架调整、平滑肌细胞收缩、神经再生、肿瘤细胞浸润、细胞凋亡的调节等一系列生物学过程。Rho/ROCK激活后可以作用于多种底物，从而产生生物学过程。最主要的两种底物是肌球蛋白轻链(MLC)和肌球蛋白轻链磷酸酶(MLCP)，MLC的磷酸化水平是决定平滑肌收缩程度的一个重要因素。肌球蛋白轻链激酶(MLCK)磷酸化MLC的Ser-19位点，导致平滑肌收缩；MLCP的抑制可以使MLC的磷酸化和平滑肌的收缩进一步增强。ROCK被激活以后，本身可以将MLC磷酸化而发生肌丝收缩作用；同时也能将MLCP磷酸化，使MLCP失活，导致细胞胞浆内MLC磷酸化程度增高，间接促进肌丝收缩。
在动物模型中Rho激酶活性的抑制展现出治疗人类疾病的多种益处，包括心血管疾病如肺动脉高压、高血压、动脉粥样硬化、心脏肥大、高眼压、脑缺血、脑血管痉挛等，和中枢神经系统病症如神经元变性等。研究表明ROCK表达和活性在自发性高血压大鼠中有所升高，说明其与这些动物高血压的发生具有关联(Involvement of Rho-kinase in hypertensive vascular disease:a novel therapeutic target in hypertension[J].FASEB J.,2001,15(6):1062-4)。ROCK抑制剂Y-27632可使三种大鼠高血压模型(自发性高血压、肾性高血压、醋酸脱氧皮质酮盐型高血压)中的血压显著降低，而对对照大鼠的血压作用较小(Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension[J].Nature,1997,389(6654):990-4)。也有研究表明ROCK抑制剂对肺动脉高压具有较好的作用(Acute vasodilator effects of a Rho-kinase inhibitor,fasudil,in patients with severe pulmonary hypertension[J].Heart,2005:91(3):391-2)。
目前已经研究开发的ROCK抑制剂可分为五大类：(1)异喹啉类：此类化合物结构特点是具有一个异喹啉结构和哌嗪环，两者通过磺酰基相连。代表物有法苏地尔(Uehata M,Ishizaki T,Satoh H,et al.Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension[J].Nature,1997,389:990-994)、H-1152P(Tamura M,Nakao H,Yoshizaki H,et al.Development of specific Rho-kinase inhibitors and their clinical application[J].Biochim Biophys Acta,2005,1754:245-252)；(2)4-氨基吡啶类： 此类化合物结构除4-氨基吡啶母核外，在分子的中心位置还含有一个环己烷或苯环结构，在环己烷的4位具有侧链结构。代表物有Y-30141(Takami A,Iwakubo M,Okada Y,et al.Design and synthesis of Rho kinase inhibitors[J].Bioorg Med Chem,2004,12:2115-2137)；(3)吲唑类：此类化合物将5-氨基或5-烷氧基-1H吲唑作为骨架；(4)酰胺和脲类：此类化合物具有一个邻苯二甲酰亚胺和一个脲基构成的绞和结构。(5)其它类：其它不包含上述结构的ROCK抑制剂，代表物有Rockout(Yarrow JC,Totsukawa G,Charras GT,et al.Screening for cell migration inhibitors via automated microscopy reveals a Rho-kinase inhibitor[J].Chem Biol,2005,12:385-395)。
目前已上市的ROCK抑制剂药物有Asahi Kasei公司的Eril(适用于脑血管痉挛的治疗)和Kowa公司的
Figure PCTCN2018091161-appb-000001
(K-115)(适用于高眼压症和青光眼的治疗)。其中
Figure PCTCN2018091161-appb-000002
仅在日本上市销售。因此进行开发作用于ROCK的靶向小分子药物研究，得到活性更好、选择性更高、作用时间更长、稳定性更高、更低毒性和副作用、更经济的ROCK抑制剂，具有十分重要的社会和经济意义。
发明内容
本发明提供了一种式(Ⅰ)所示的化合物或其立体异构体：
Figure PCTCN2018091161-appb-000003
其中，
n分别为0、1或2；
R 1分别独立地选自氢、羟基、卤素、氨基、羧基、三氟甲基、硝基、氰基、C 1～6烷基；
m分别为0、1、2、3、4或5；
R 3选自-NR 2R 2’、含N的杂环烷基；其中，含N的杂环烷基可以进一步被1～2个独立选自于卤素、C 1～6烷基的取代基取代；
R 2、R 2’分别独立选自氢、C 1～6烷基；
A环选自5～6元芳环、5～6元芳杂环或
Figure PCTCN2018091161-appb-000004
B环、C环分别独自选自5～6元芳环、5～6元芳杂环；
R 4分别独立地选自氢、卤素、硝基、氰基、C 1～6烷基、C 2～6烯基、C 2～6炔基、3～6元环烷基、3～6元杂环烷基、-(CH 2) mOR a、-(CH 2) mOC(O)R a、-(CH 2) mOC(O)NR aR b、-(CH 2) mNR aR b、-(CH 2) mNR aC(O)R b、-(CH 2) mNR aC(O)OR b、-(CH 2) mC(O)R a、-(CH 2) mC(O)OR a、-(CH 2) mC(O)NR aR b；
R a、R b分别独自选自氢、C 1～6烷基、C 2～6烯基、C 2～6炔基、3～6元环烷基、3～6元杂环烷基、5～6元芳环、5～6元芳杂环；其中，3～6元环烷基、3～6元杂环烷基、5～6元芳环、5～6元芳杂环可以进一步被1～2个独立选自于卤素、C 1～6烷基的取代基取代。
优选的，所述R 3选自的含N的杂环烷基为4～5元的含N杂环烷基；
更进一步的，当R 3选自含N的杂环烷基且A环有2个R 4取代时，2个R 4不同时为卤素；
本发明还提供一种结构如式(Ⅱ)所示的化合物或其立体异构体：
Figure PCTCN2018091161-appb-000005
其中，
R 1选自氢、羟基、卤素、氨基、羧基、三氟甲基、硝基、氰基、C 1～6烷基；
m分别为0、1、2或3；
R 2、R 2’分别独立选自氢、C 1～6烷基；
A环选自5～6元芳环、5～6元芳杂环或
Figure PCTCN2018091161-appb-000006
B环、C环分别独自选自5～6元芳环、5～6元芳杂环；
R 4分别独立地选自氢、卤素、硝基、氰基、C 1～6烷基、C 2～6烯基、C 2～6炔基、3～6元环烷基、3～6元杂环烷基、-(CH 2) mOR a、-(CH 2) mOC(O)R a、-(CH 2) mNR aR b、-(CH 2) mNR aC(O)R b、-(CH 2) mC(O)R a、-(CH 2) mC(O)OR a、-(CH 2) mC(O)NR aR b；
R a、R b分别独自选自氢、C 1～6烷基、C 2～6烯基、C 2～6炔基、3～6元环烷基、3～6元杂环烷基、5～6元芳环、5～6元芳杂环；其中，3～6元环烷基、3～6元杂环烷基、5～6元芳环、5～6元芳杂环可以进一步被1～2个独立选自于卤素、C 1～6烷基的取代基取代。
优选的，式(Ⅱ)的化合物如下列结构式所示：
Figure PCTCN2018091161-appb-000007
Figure PCTCN2018091161-appb-000008
另一方面，本发明还提供结构如式(Ⅲ)所示的化合物或其立体异构体：
Figure PCTCN2018091161-appb-000009
R 1选自氢、羟基、卤素、氨基、羧基、三氟甲基、硝基、氰基、C 1～6烷基；
m分别为0、1、2或3；
A环选自5～6元芳环、5～6元芳杂环或
Figure PCTCN2018091161-appb-000010
B环、C环分别独自选自5～6元芳环、5～6元芳杂环；
E环选自含N的杂环烷基；所述含N的杂环烷基可以进一步被1～2个独立选自于卤素、C 1～6烷基的取代基取代；
R 4分别独立地选自氢、卤素、硝基、氰基、C 1～6烷基、C 2～6烯基、C 2～6炔基、3～6元环烷基、3～6元杂环烷基、-(CH 2) mOR a、-(CH 2) mOC(O)R a、-(CH 2) mNR aR b、-(CH 2) mNR aC(O)R b、-(CH 2) mC(O)R a、-(CH 2) mC(O)OR a、-(CH 2) mC(O)NR aR b；
R a、R b分别独自选自氢、C 1～6烷基、C 2～6烯基、C 2～6炔基、3～6元环烷基、3～6元杂环烷基、5～6元芳环、5～6元芳杂环；其中，3～6元环烷基、3～6元杂环烷基、5～6元芳环、5～6元芳杂环可以进一步被1～2个独立选自于卤素、C 1～6烷基的取代基取代。
优选的，E环选自4～5元的含N的杂环烷基。
更优选的，式(Ⅲ)化合物如下列结构所示：
Figure PCTCN2018091161-appb-000011
本发明的再一方面，还提供上述化合物或其立体异构体、晶型、药学上可接受的盐、水合物或溶剂合物在制备治疗与ROCK活性异常相关的疾病的药物中的用途。
进一步的，所述与ROCK活性异常相关的疾病是细胞骨架调整、平滑肌细胞收缩、神经再生相关的疾病。
更进一步的，所述的疾病为高眼压症或者青光眼。
本发明的另一方面，还提供一种药物，它是以上述化合物或其立体异构体、晶型、药学上可接受的盐、水合物或溶剂合物为活性成分，加上药学上可接受的辅料制备而成的制剂。
本发明中提供的化合物和衍生物可以根据IUPAC(国际纯粹与应用化学联合会)或CAS(化学文摘服务社，Columbus，OH)命名系统命名。
关于本发明的使用术语的定义：除非另有说明，本文中基团或者术语提供的初始定义适用于整篇说明书的该基团或者术语；对于本文没有具体定义的术语，应该根据公开内容和上下文，给出本领域技术人员能够给予它们的含义。
“取代”是指分子中的氢原子被其它不同的原子或分子所替换。
碳氢基团中碳原子含量的最小值和最大值通过前缀表示，例如，前缀C a～ b烷基表明任何含“a”至“b”个碳原子的烷基。因此，例如，C 1～ 4烷基是指包含1～4个碳原子的烷基。
本发明中烯基可含有一个或多个碳碳双键。本发明中炔基可含有一个或多个碳碳三 键。
______________________________________________________________________________
本发明中C a～ b烷氧基、C a～ b烷酯基、C a～ b烷氨基、C a～ b酰基分别是指含有“a”至“b”个碳原子的烷基与对应的氧原子、酯基、氨基、酰基相连得到的基团。
本发明中的立体异构体包括对映异构体和非对映异构体。
本发明中卤素包括氟原子、氯原子、溴原子、碘原子等。
本发明中杂原子包括氮原子、氧原子、硫原子等。
本发明中环烷基指非芳香性的环状碳氢基团，包括饱和环烷基和部分饱和环烷基。
本发明中杂环烷基指环中含有杂原子的非芳香性环烷基，包括饱和杂环烷基和部分饱和杂环烷基。
本发明中芳环指具有芳香性的环状碳氢基团。
本发明中芳杂环指环中含有杂原子的芳环。
术语“药学上可接受的”是指某载体、运载物、稀释剂、辅料，和/或所形成的盐通常在化学上或物理上与构成某药物剂型的其它成分相兼容，并在生理上与受体相兼容。
术语“盐”和“可药用的盐”是指上述化合物或其立体异构体，与无机和/或有机酸和碱形成的酸式和/或碱式盐，也包括两性离子盐(内盐)，还包括季铵盐，例如烷基铵盐。这些盐可以是在化合物的最后分离和纯化中直接得到。也可以是通过将上述化合物，或其立体异构体，与一定数量的酸或碱适当(例如等当量)进行混合而得到。这些盐可能在溶液中形成沉淀而以过滤方法收集，或在溶剂蒸发后回收而得到，或在水介质中反应后冷冻干燥制得。本发明中所述盐可以是化合物的盐酸盐、硫酸盐、枸橼酸盐、苯磺酸盐、氢溴酸盐、氢氟酸盐、磷酸盐、乙酸盐、丙酸盐、丁二酸盐、草酸盐、苹果酸盐、琥珀酸盐、富马酸盐、马来酸盐、酒石酸盐或三氟乙酸盐。
在某些实施方式中，本发明的一种或多种化合物可以彼此联合使用。也可选择将本发明的化合物与任何其它的活性试剂结合使用，用于制备调控细胞功能或治疗疾病的药物或药物组合物。如果使用的是一组化合物，则可将这些化合物同时、分别或有序地对受试对象进行给药
显然，根据本发明的上述内容，按照本领域的普通技术知识和惯用手段，在不脱离本发明上述基本技术思想前提下，还可以做出其它多种形式的修改、替换或变更。
以下通过实施例形式的具体实施方式，对本发明的上述内容再作进一步的详细说明。但不应将此理解为本发明上述主题的范围仅限于以下的实例。凡基于本发明上述内容所实现的技术均属于本发明的范围。
附图说明
图1为K115对正常眼压新西兰白兔的降眼压活性的实验结果
图2为化合物5对正常眼压新西兰白兔的降眼压活性的实验结果
图3为化合物11a对正常眼压新西兰白兔的降眼压活性的实验结果
图4为化合物11a对高眼压新西兰白兔的降眼压活性的实验结果
具体实施方式
化合物结构通过核磁共振(NMR)和质谱(MS)来确定的。NMR位移(δ)以10-6(ppm)的单位给出。NMR的测定是用(Bruker AvanceIII 400和Bruker Avance 300)核磁仪，测定溶剂为氘代二甲基亚砜(DMSO-d6)、氘代氯仿(CDCl 3)、氘代甲醇(MeOD)，内标为四甲基硅烷(TMS)。
LC-MS的测定使用岛津液质联用仪(Shimadzu LC-MS 2020(ESI))。
HPLC的测定使用岛津高压液相色谱仪(Shimadzu LC-20A)。
MPLC(中压制备色谱)使用Gilson GX-281反相制备色谱仪。
柱层析一般使用烟台黄海硅胶200～300目硅胶为载体。
薄层层析硅胶板用烟台黄海HSGF254或青岛GF254硅胶板，薄层层析分离纯化产品采用的规格是0.4mm～0.5mm。
本发明具体实施方式中使用的原料、设备均为已知产品，通过购买市售产品获得。已知的起始原料可以采用或按照本领域已知的方法来合成，或可购买于安耐吉化学、成都科龙化工、韶远化学科技、百灵威科技等公司。
室温为最适宜的反应温度，为20℃～30℃。实施例中无特殊说明，M是摩尔每升。实施例中无特殊说明，溶液是指水溶液。
简写说明：DCM为二氯甲烷；EA、EtOAc为乙酸乙酯；PE为石油醚；THF为四氢呋喃；DMF为N,N-二甲基甲酰胺；DIEA为二异丙基乙胺；DMAP为4-二甲氨基吡啶；EDCI为1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐；HOBT为1-羟基苯并三唑；HBTU为苯并三氮唑-N,N,N',N'-四甲基脲六氟磷酸盐；Pd(dppf)Cl 2为[1,1'-双(二苯基膦基)二茂铁]二氯化钯。
实施例1、N-(3-氨基-1-苯基丙基)异喹啉-6-甲酰胺的制备。
步骤1.(S)-N-苯亚甲基-2-甲基丙烷-2-磺酰胺
Figure PCTCN2018091161-appb-000012
室温下向苯甲醛(30.0g,283mmol)的四氢呋喃(150mL)溶液中加入叔丁基亚磺酰胺(51.4g,424mmol)和钛酸四异丙酯(100g,353mmol)，于60℃搅拌4小时后加水过滤，用乙酸乙酯萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得(S)-N-苯亚甲基-2-甲基丙烷-2-磺酰胺(54g，232mmol，产率82％)。
MS(ESI)m/z＝210(M+1) +。
步骤2.(S)-2-甲基-N-(2-硝基-1-苯乙基)-丙烷-2-亚磺酰胺的制备
Figure PCTCN2018091161-appb-000013
(S)-N-苯亚甲基-2-甲基丙烷-2-磺酰胺(48.0g，229mmol)溶于四氢呋喃(500mL)，在氮气保护下于0℃加入叔丁醇钾(38.5g，344mmol)，反应液搅拌1小时后加入硝基甲烷(140g，2.29mol)，室温搅拌24小时。反应液用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得到(S)-2-甲基-N-(2-硝基-1-苯乙基)-丙烷-2-亚磺酰胺(18g，66.6mmol，产率29％)。
MS(ESI)m/z＝271(M+1) +。
步骤3.(S)-2-甲基-N-(2-胺基-1-苯乙基)-丙烷-2-亚磺酰胺的制备
Figure PCTCN2018091161-appb-000014
将(S)-2-甲基-N-(2-硝基-1-苯乙基)-丙烷-2-亚磺酰胺(8.00g，29.6mmol)溶于甲醇(50.0mL)，加入雷尼镍(800mg，10％)，室温搅拌24小时。反应液用硅藻土过滤，减压蒸除溶剂得(S)-2-甲基-N-(2-胺基-1-苯乙基)-丙烷-2-亚磺酰胺(6.00g，21.5mmol，产率86％)。
MS(ESI)m/z＝241(M+1) +。
步骤4.苄基-N-(((S)-叔丁基亚磺酰胺)-2-苯乙基)氨基甲酸酯的制备
Figure PCTCN2018091161-appb-000015
将(S)-2-甲基-N-(2-胺基-1-苯乙基)-丙烷-2-亚磺酰胺(7.00g，29.1mmol)溶于四氢呋喃(70.0mL)，加入三乙胺(11.8g，116mmol)和苯甲氧羰酰琥珀酰亚胺(8.70g，34.9mmol)，搅拌1小时后用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得苄基-N-(((S)-叔丁基亚磺酰胺)-2-苯乙基)氨基甲酸酯(6.02g，12.8mmol，产率44％)。
MS(ESI)m/z＝375(M+1) +。
步骤5.苄基-N-(2-氨基-2-苯基-乙基)氨基甲酸酯的制备
Figure PCTCN2018091161-appb-000016
将苄基-N-(((S)-叔丁基亚磺酰胺)-2-苯乙基)氨基甲酸酯(5.00g，13.4mmol)溶于甲醇(10.0mL)，加入1,4-二氧六环的盐酸溶液(40.0ml，5mol/L)，搅拌1小时后减压蒸除溶剂得苄基-N-(2-氨基-2-苯基-乙基)氨基甲酸酯(3.01g，10.2mmol，产率76％)。
MS(ESI)m/z＝271(M+1) +。
步骤6.苄基-N-((2S)-2-(异喹啉-6-甲酰胺)-2-苯基-乙基)氨基甲酸酯的制备
Figure PCTCN2018091161-appb-000017
将苄基-N-(2-氨基-2-苯基-乙基)氨基甲酸酯(2.00g，7.40mmol)溶于DMF(10.0mL)，加入苯并三氮唑-N,N,N',N'-四甲基脲六氟磷酸酯(2.07g，8.14mmol),异喹啉-6-甲酸(1.28g，7.40mmol)和N,N-二异丙基乙胺(3.38g，29.6mmol)，搅拌1小时后用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得苄基-N-((2S)-2-(异喹啉-6-甲酰胺)-2-苯基-乙基)氨基甲酸酯(2.01g，4.24mmol，产率57％)。
MS(ESI)m/z＝426(M+1) +。
步骤7.N-((S)-2氨基-1苯基-乙基)异喹啉-6-甲酰胺的制备
Figure PCTCN2018091161-appb-000018
将苄基-N-((S)-2-(异喹啉-6-甲酰胺)-2-苯基-乙基)氨基甲酸酯(300mg，0.71mmol)溶于乙酸(5.00mL)，加入氢溴酸醋酸溶液(2.50ml，33％)，搅拌1小时后减压蒸除溶剂得N-((S)-2氨基-1苯基-乙基)异喹啉-6-甲酰胺(180mg，540μmol，产率76％)。
MS(ESI)m/z＝292(M+1) +。
1H NMR(400MHz,DMSO-d 6):δ＝9.78-9.84(m，2H),8.94(s,1H),8.73-8.74(d,J＝4,1H),8.52-8.54(d,J＝8Hz,1H),8.39-8.46(m,4H),7.50-7.52(d,J＝8,2H),7.32-7.42(m,3H),5.41-5.46(m,1H),3.44-3.49(m,1H),3.17-3.24(m,1H)。
实施例2、(S)-N-(3-氨基-1-(3-溴苯基)丙基)异喹啉-6-甲酰胺的制备
Figure PCTCN2018091161-appb-000019
按照实施例1中步骤1到步骤7的方法，并将步骤1中的苯甲醛换为3-溴苯甲醛，得到(S)-N-(3-溴苯亚甲基)-2-甲基-叔丁基亚磺酰胺。
MS(ESI)m/z＝370,372(M+1) +。
1HNMR(400MHz,MeOD):δ＝9.91(s,1H),8.96(s,1H),8.65-8.71(m,3H),8.49-8.52(J＝12Hz,1H),7.78(t,J＝7.2Hz,1H),7.54-7.57(m,2H),7.36-7.42(m,1H),3.64-3.75(m,1H),1.87-1.81(m,2H)。
实施例3、(S)-N-(2-氨基-1-(4-(羟甲基)苯基)乙基)异喹啉-6-甲酰胺的制备
步骤1.(S)-4-(((叔丁基亚磺酰基)亚胺)甲基)苯甲酸异丙酯的制备
Figure PCTCN2018091161-appb-000020
室温下向(S)-叔丁基亚磺酰胺(83.1g，685mmol)的THF(50.0mL)溶液中加入4-甲酰基苯甲酸甲酯(50.0g，305mmol)和四异丙基钛(108g，381mmol)，回流搅拌2小时。待反应液冷却后用水(200ml)淬灭，滤出白色固体。滤液分层，水相用乙酸乙酯萃取(200mL×1)。合并有机相依次饱和食盐水(300mL×1)、水(300mL×1)洗各1次后用无水Na 2SO 4干燥，过滤，滤液减压蒸除溶剂。柱层析纯化得(S，E)-4-(((叔丁基亚磺酰基)亚胺)甲基)苯甲酸异丙酯(60g，203mmol，产率67％)。
MS(ESI)m/z＝296(M+1) +。
步骤2. 4-((S)-1-((S)-1,1-二甲基乙基亚磺酰胺)-2-硝基乙基)苯甲酸异丙酯的制备
Figure PCTCN2018091161-appb-000021
在冰浴下往硝基甲烷(31.0g，654mmol)的THF(400mL)溶液中分批加入叔丁醇钾(11.0g，98.1mmol)，氮气保护下室温搅拌30分钟。于冰浴下滴入(S)-4-(((叔丁基亚磺酰基)亚胺)甲基)苯甲酸异丙酯(18.0g，65.4mmol)的THF(200mL)溶液，氮气保护下室温搅拌过夜。反应液用水淬灭(400mL)，乙酸乙酯萃取(400mL)。有机相依次用饱和食盐水(400mL)洗1次后用无水Na 2SO 4干燥，过滤，滤液减压蒸除溶剂。柱层析纯化得4-((S)-1-((S)-1,1-二甲基乙基亚磺酰胺)-2-硝基乙基)苯甲酸异丙酯(12g，35.7mmol，产率55％)。
MS(ESI)m/z＝357(M+1) +。
步骤3. 4-((S)-2-氨基-1-((S)-1,1-二甲基乙基亚磺酰胺)乙基)苯甲酸异丙酯的制备
Figure PCTCN2018091161-appb-000022
氮气保护下于室温往4-((S)-1-((S)-1,1-二甲基乙基亚磺酰胺)-2-硝基乙基)苯甲酸异丙酯(10.0g，28.1mmol)的甲醇(30.0mL)溶液中加入雷尼镍(1.00g，10％w/w)和三乙胺(1.42g，14.0mmol)。于氢气下室温搅拌4小时。反应液过滤，滤饼甲醇洗涤，滤液减压蒸除溶剂得4-((S)-2-氨基-1-((S)-1,1-二甲基乙基亚磺酰胺)乙基)苯甲酸异丙酯(9.40g，26.9mmol，产率96％)。
MS(ESI)m/z＝327(M+1) +。
步骤4.(S)-N-((S)-2-氨基-1-(4-(羟甲基)苯基)乙基)-2-甲基丙烷-2-亚磺酰胺的制备
Figure PCTCN2018091161-appb-000023
氮气保护下于室温往4-((S)-2-氨基-1-((S)-1,1-二甲基乙基亚磺酰胺)乙基)苯甲酸异丙酯(8.00g，24.5mmol)的THF(200mL)溶液中加入LiBH 4(4.50g，204mmol)。于氮气下回流16小时。反应液于冰浴下用水淬灭(300mL)，EtOAc萃取(300mL×2)。有机相用无水Na 2SO 4干燥，过滤，滤液减压蒸除溶剂得(S)-N-((S)-2-氨基-1-(4-(羟甲基)苯基)乙基)-2-甲基丙烷-2-亚磺酰胺(4.6g，15.7mmol，产率77％)。
MS(ESI)m/z＝271(M+1) +。
步骤5.(9H-芴-9-基)甲基-((S)-2-((S)-1,1-二甲基乙基亚磺酰胺)-2-(4-(羟甲基)苯基)乙基)氨基甲酸酯的制备
Figure PCTCN2018091161-appb-000024
室温下往(S)-N-((S)-2-氨基-1-(4-(羟甲基)苯基)乙基)-2-甲基丙烷-2-亚磺酰胺(3.97g，14.7mmol)的THF(20mL)和水(20mL)的溶液中加入(9H-芴-9-基)甲基-(2,5-二酮吡咯烷-1-基)碳酸酯(7.44g，22.1mmol)和NaHCO 3(3.70g，44.1mmol)。于室温下搅拌1小时。用乙酸乙酯萃取(50mL×2)，合并有机相后用饱和食盐水(50mL×1)洗1次，有机相用无水 Na 2SO 4干燥，过滤，滤液减压蒸除溶剂得(9H-芴-9-基)甲基((S)-2-((S)-1,1-二甲基乙基亚磺酰胺)-2-(4-(羟甲基)苯基)乙基)氨基甲酸酯(5.00g，9.70mmol，产率66％)。
MS(ESI)m/z＝493(M+1) +。
步骤6.(S)-(9H-芴-9-基)甲基(2-氨基-2-(4-(羟基)苯基)乙基)氨基甲酸酯盐酸盐的制备
Figure PCTCN2018091161-appb-000025
室温下往(9H-芴-9-基)甲基((S)-2-((S)-1,1-二甲基乙基亚磺酰胺)-2-(4-(羟甲基)苯基)乙基)氨基甲酸酯(2.30g，4.67mmol)的甲醇(3.0mL)溶液中加入饱和盐酸二氧六环溶液(30mL)。于室温下搅拌30分钟。反应液减压蒸除溶剂得(S)-(9H-芴-9-基)甲基(2-氨基-2-(4-(羟基)苯基)乙基)氨基甲酸酯盐酸盐(1.90g，4.47mmol，产率96％)。
MS(ESI)m/z＝389(M+1) +。
步骤7.(S)-(9H-芴-9-基)甲基(2-(4-(羟甲基)苯基)-2-(异喹啉-6-甲酰胺)乙基)氨基甲酸酯的制备
Figure PCTCN2018091161-appb-000026
室温下往异喹啉-6-甲酸(293mg，1.69mmol)和(S)-(9H-芴-9-基)甲基(2-氨基-2-(4-(羟基)苯基)乙基)氨基甲酸酯盐酸盐(600mg，1.41mmol)的DMF(10.0mL)溶液中加入EDCI(325mg，1.69mmol)，HOBT(229mg，1.69mmol)和DMAP(17.2mg，0.14mmol)。于室温下搅拌2小时。用水淬灭(30mL)，EtOAc萃取(30mL×2)。合并有机相用饱和食盐水(30mL×1)洗1次，有机相用无水Na 2SO 4干燥，过滤，滤液减压蒸除溶剂。柱层析纯化得(S)-(9H-芴-9-基)甲基(2-(4-(羟甲基)苯基)-2-(异喹啉-6-甲酰胺)乙基)氨基甲酸酯(497mg，0.86mmol，产率61％)。
MS(ESI)m/z＝544(M+1) +。
步骤8.(S)-N-(2-氨基-1-(4-(羟甲基)苯基)乙基)异喹啉-6-甲酰胺的制备
Figure PCTCN2018091161-appb-000027
室温下往(S)-(9H-芴-9-基)甲基(2-(4-(羟甲基)苯基)-2-(异喹啉-6-甲酰胺)乙基)氨基甲酸酯(140mg，0.258mmol)的THF(15.0mL)溶液中加入哌啶(219mg，2.58mmol)。于室温下搅拌1小时。反应液减压蒸除溶剂，MPLC纯化得(S)-N-(2-氨基-1-(4-(羟甲基)苯基)乙基)异喹啉-6-甲酰胺(38.4mg，92.4μmol，产率95％)。
MS(ESI)m/z＝322(M+1) +。
1H NMR(400MHz,MeOD):δ＝9.91(s,1H),8.90(d,J＝16.48Hz,1H),8.71(d,J＝6.8Hz,1H),8.66(s,1H),8.64(s,1H),8.48(dd,J＝1.52Hz,J＝1.60Hz,1H),7.56(s,1H),7.54(s,1H),7.47(s,1H),7.45(s,1H),5.57(q,1H),4.67(q,2H),3.62(t,1H),3.49(t,1H)。
实施例4、(S)-4-(2-氨基-1-(异喹啉-6-甲酰胺基)乙基)苯基2,4-二甲基苯甲酸酯的制备
步骤1.(S)-4-(2-((((9H-芴-9-基)甲氧基)羰基)氨基)-1-(异喹啉-6-甲酰胺基)乙基)苯基2,4-二甲基苯甲酸酯
Figure PCTCN2018091161-appb-000028
室温下往(S)-(9H-芴-9-基)甲基(2-(4-(羟甲基)苯基)-2-(异喹啉-6-甲酰胺)乙基)氨基甲酸酯(158mg，281μmol)和2,4-二甲基苯甲酸(50.7mg，328μmol)的二氯甲烷溶液(40.0mL)中加入EDCI(70.2mg，366μmol)和DMAP(3.44mg，128μmol)，室温搅拌过夜。减压蒸除溶剂，柱层析纯化得(S)-4-(2-((((9H-芴-9-基)甲氧基)羰基)氨基)-1-(异喹啉-6-甲酰胺基)乙基)苯基2,4-二甲基苯甲酸酯(151mg，0.22mmol，产率77％)。
MS(ESI)m/z＝676(M+1) +。
步骤2.(S)-4-(2-氨基-1-(异喹啉-6-甲酰胺基)乙基)苯基2,4-二甲基苯甲酸酯的制备
Figure PCTCN2018091161-appb-000029
室温下往(S)-4-(2-((((9H-芴-9-基)甲氧基)羰基)氨基)-1-(异喹啉-6-甲酰胺基)乙基)苯基2,4-二甲基苯甲酸酯(150mg，222μmol)的THF(10.0mL)溶液中加入哌啶(378mg，4.44mmol)。于室温下搅拌过夜。减压蒸除溶剂后经MPLC纯化得(S)-4-(2-氨基-1-(异喹啉-6-甲酰胺基)乙基)苯基-2,4-二甲基苯甲酸酯(35mg，71.4μmol，产率14％)。
MS(ESI)m/z＝454(M+1) +。
1H NMR(400MHz,MeOD):δ＝9.89(s,1H),8.88(s,1H),8.70(d,J＝6.52Hz,1H),8.65(d,J＝3.24Hz,1H),8.63(s,1H),8.48(dd,J＝1.48Hz,J＝1.52Hz,1H),7.83(d,J＝7.96Hz,1H), 7.59(q,4H),7.12(s,1H),7.09(d,J＝8.0Hz,1H),5.59(q,1H),5.36(s,2H),3.63(q,1H),3.50(dd,J＝4.92Hz,J＝4.52Hz,1H),2.55(s,3H),2.36(s,3H)。
实施例5、N-((1S)-2-(二甲氨基)-1-苯基-乙基)异喹啉-6-甲酰胺的制备
步骤1.N-((S)-2-(二甲氨基)-1-苯基-乙基)-2-甲基-丙烷-2-亚磺酰胺的制备
Figure PCTCN2018091161-appb-000030
将(S)-2-甲基-N-(2-胺基-1-苯乙基)-丙烷-2-亚磺酰胺(700mg，2.96mmol)溶于甲醇(70mL)，加入乙酸(178mg，2.96mmol)和甲醛水溶液(266mg，8.88mmol)，搅拌1小时后加入氰基硼氢化钠(744mg，11.8mmol)，继续反应2小时后用EA和水萃取，再将水相用EA萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得N-((1S)-2-(二甲氨基)-1-苯基-乙基)-2-甲基-丙烷-2-亚磺酰胺(201mg，0.59mmol，产率20％)。
MS(ESI)m/z＝269(M+1) +。
步骤2.(1S)-N’,N’-二甲基-1-苯基-乙烷-1,2-联氨的制备
Figure PCTCN2018091161-appb-000031
将N-((1S)-2-(二甲氨基)-1-苯基-乙基)-2-甲基-丙烷-2-亚磺酰胺(200mg，0.75mmol)溶于甲醇(5.0mL)，加入盐酸二氧六环溶液(10ml，5M)搅拌1小时后减压蒸除溶剂得(1S)-N’,N’-二甲基-1-苯基-乙烷-1,2-联氨(100mg，0.48mmol，产率65％)。
MS(ESI)m/z＝165(M+1) +。
步骤3.N-((1S)-2-(二甲氨基)-1-苯基-乙基)异喹啉-6-甲酰胺的制备
Figure PCTCN2018091161-appb-000032
将(1S)-N’,N’-二甲基-1-苯基-乙烷-1,2-联氨(90.0mg，550μmol)溶于DMF(10.0mL)，加入苯并三氮唑-N,N,N',N'-四甲基脲六氟磷酸酯(153mg，600μmol)、异喹啉-6-甲酸(105 mg，600μmol)和N,N-二异丙基乙胺(284g，2.19mmol)，搅拌1小时后用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得苄基-N-((2S)-2-(异喹啉-6-甲酰胺)-2-苯基-乙基)氨基甲酸酯(52mg，0.13mmol，产率24％)。
MS(ESI)m/z＝320(M+1) +。
1H NMR(400MHz,DMSO-d 6):δ＝10.11(s,1H),10.00-10.3(d,J＝8.4,1H),9.88(s,1H),9.02(s,1H),8.74-8.75(d,J＝6.4,1H),8.46-8.56(m,3H),7.59-7.61(m,2H),7.34-7.43(m,3H),5.62-5.66(m,1H),3.86-3.92(m,1H),3.45-3.49(m,1H),2.88-2.92(m,6H)。
实施例6、N-(3-氨基-1-(3-氯苯基)丙基)异喹啉-6-甲酰胺的制备。
步骤1. 3-氨基-3-(3-氯苯基)丙烯腈的制备
Figure PCTCN2018091161-appb-000033
室温下往3-氯苯腈(8.00g，58.2mmol)的甲苯(50.0mL)溶液中加入乙腈(4.77g，116mmol)和叔丁醇钾(15.0g，134mmol)并且搅拌6小时，用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得到3-氨基-3-(3-氯苯基)丙烯腈(4.99g，28.0mmol，产率48％)。
MS(ESI)m/z＝179(M+1) +。
步骤2. 3-氨基-3-(3-氯苯基)丙腈的制备
Figure PCTCN2018091161-appb-000034
将3-氨基-3-(3-氯苯基)丙烯腈(5.00g，28.0mmol)溶于乙醇(10.0mL)，室温下加入氰基硼氢化钠(2.11g，33.6mmol)和1滴0.5％的溴甲酚绿乙醇溶液。然后加入浓盐酸直到反应液保持黄色，继续室温搅拌3小时。减压蒸除溶剂并用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得到3-氨基-3-(3-氯苯基)丙腈(5.02g，27.7mmol，产率99％)。
MS(ESI)m/z＝181(M+1) +。
步骤3.N-(1-(3-氯苯基)-2-氰乙基)异喹啉-6-甲酰胺的制备
Figure PCTCN2018091161-appb-000035
将6-异喹啉甲酸(1.50g，8.30mmol)溶于DMF(15.0mL)，室温下加入HBTU(3.47g，9.13mmol)、DIEA(3.21g，24.9mmol)和3-氨基-3-(3-氯苯基)丙腈(1.58g，9.13mmol)。室温下搅拌3小时。减压蒸除溶剂并用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得N-(1-(3-氯苯基)-2-氰乙基)异喹啉-6-甲酰胺(1.8g，5.36mmol，产率65％)。
MS(ESI)m/z＝336(M+1) +。
步骤4.N-(3-氨基-1-(3-氯苯基)丙基)异喹啉-6-甲酰胺的制备
Figure PCTCN2018091161-appb-000036
将N-(1-(3-氯苯基)-2-氰乙基)异喹啉-6-甲酰胺(70.0mg，210μmol)溶于甲醇(5.00mL)，室温下加入雷尼镍(10.0mg)。在氢气下于室温搅拌3小时。过滤除去固体，减压蒸除溶剂。经MPLC纯化得N-(3-氨基-1-(3-氯苯基)丙基)异喹啉-6-甲酰胺(15.2mg，42.0μmol，产率20％)。
MS(ESI)m/z＝340(M+1) +。
1H NMR(400MHz,MeOD):δ＝9.34(s,1H),8.56-8.54(m,1H),8.44(s,1H),8.25-8.23(m,1H),8.10-8.08(m,1H),7.98-7.96(m,1H),7.53-7.52(m,1H),7.44-7.41(m,1H),7.39-7.37(m,1H),7.34-7.31(m,1H),5.31-5.27(m,1H),2.92-2.78(m,2H),2.24-2.06(m,2H)。
实施例7、4-(3-(3-氨基-1-(异喹啉-6-甲酰胺)丙基)苯基)噻吩-2-羧酸乙酯的制备
步骤1.叔丁基(1-(3-氯苯基)-2-氰乙基)氨基甲酸酯的制备
Figure PCTCN2018091161-appb-000037
将3-氨基-3-(3-氯苯基)丙腈(5.00g，27.7mmol)溶于THF/H 2O/MeOH＝4/1/1(100.00mL)的混合溶液中，室温下加入碳酸钠(8.80g，83.0mmol)和碳酸二叔丁酯(6.65g，30.5mmol)。室温搅拌3小时。减压蒸除溶剂并用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次， 合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得叔丁基(1-(3-氯苯基)-2-氰乙基)氨基甲酸酯(5.6g，20mmol，产率72％)。
MS(ESI)m/z＝281(M+1) +。
步骤2.叔丁基(3-氨基-1-(3-氯苯基)丙基)氨基甲酸甲酯的制备
Figure PCTCN2018091161-appb-000038
将叔丁基(1-(3-氯苯基)-2-氰乙基)氨基甲酸酯(2.00g，7.12mmol)溶于甲醇(25.0mL)，室温下加入雷尼镍(200mg)，在氢气下于室温搅拌3小时。过滤除去固体，减压蒸除溶剂。经MPLC纯化得到叔丁基(3-氨基-1-(3-氯苯基)丙基)氨基甲酸甲酯(2.01g，7.02mmol，产率99％)。
MS(ESI)m/z＝285(M+1) +。
步骤3.苄基叔丁基(1-(3-氯苯基)丙烷-1,3-取代基)二氨基甲酸酯的制备
Figure PCTCN2018091161-appb-000039
将叔丁基(3-氨基-1-(3-氯苯基)丙基)氨基甲酸甲酯(2.00g，7.02mmol)溶于四氢呋喃(50.0mL)，室温下加入三乙胺(2.13g，21.1mmol)。室温搅拌3小时。减压蒸除溶剂并用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得到苄基叔丁基(1-(3-氯苯基)丙烷-1,3-取代基)二氨基甲酸酯(2.81g，6.68mmol，产率95％)。
MS(ESI)m/z＝419(M+1) +。
步骤4.苄基叔丁基(1-(3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)丙烷-1,3-取代基)二氨基甲酸酯的制备
Figure PCTCN2018091161-appb-000040
将苄基叔丁基(1-(3-氯苯基)丙烷-1,3-取代基)二氨基甲酸酯(2.20g，5.25mmol)溶于1,4-二氧六环(30.0mL)，室温下加入联硼酸频那醇酯(5.34g，421mmol)、2-双环己基膦 -2',6'-二甲氧基联苯(43.1mg，110μmol)、醋酸钯(118mg，530μmol)和醋酸钾(1.54g，15.8mmol)。氮气保护下于100℃搅拌4小时。减压蒸除溶剂并用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得到苄基叔丁基(1-(3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)丙烷-1,3-取代基)二氨基甲酸酯(1.5g，2.94mmol，产率56％)。
MS(ESI)m/z＝511(M+1) +。
步骤5. 4-(3-(3-(苄氧甲酰胺)-1-(叔丁氧羰酰胺)丙基)苯基)噻吩-2-甲酸乙酯的制备
Figure PCTCN2018091161-appb-000041
将苄基叔丁基(1-(3-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯基)丙烷-1,3-取代基)二氨基甲酸酯(1.20g，2.35mmol)溶于1,4-二氧六环(15.0mL)，室温下加入Pd(dppf)Cl 2(172mg，240μmol)、碳酸钾(975mg，7.05mmol)和4-溴噻吩-2-羧酸乙酯(608mg，2.59mmol)。氮气保护下于100℃搅拌4小时。减压蒸除溶剂并用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得到4-(3-(3-(苄氧甲酰胺)-1-(叔丁氧羰酰胺)丙基)苯基)噻吩-2-甲酸乙酯(1.1g，2.04mmol，产率87％)。
MS(ESI)m/z＝539(M+1) +。
步骤6. 4-(3-(1-氨基-3-(苄氧甲酰胺)丙基)苯基)噻吩-2-羧酸乙酯的制备
Figure PCTCN2018091161-appb-000042
将4-(3-(3-(苄氧甲酰胺)-1-(叔丁氧羰酰胺)丙基)苯基)噻吩-2-甲酸乙酯(1.10g，2.04mmol)溶于甲醇(10.0mL)，室温下加入浓盐酸(5.00ml)。室温搅拌3小时。减压蒸除溶剂。经MPLC纯化得4-(3-(1-氨基-3-(苄甲氧酰胺)丙基)苯基)噻吩-2-羧酸乙酯(864mg，1.97mmol，产率97％)。
MS(ESI)m/z＝439(M+1) +。
步骤7. 4-(3-(3-(苄氧甲酰胺)-1-(异喹啉-6-甲酰胺)丙基)苯基)噻吩-2-羧酸乙酯的制备
Figure PCTCN2018091161-appb-000043
将6-异喹啉甲酸(390mg，2.26mmol)溶于DMF(15.0mL)，室温下加入HBTU(573mg，2.26mmol)、DIEA(794mg，6.16mmol)和4-(3-(1-氨基-3-(苄甲氧酰胺)丙基)苯基)噻吩-2-羧酸乙酯(900mg，2.05mmol)。室温搅拌3小时。减压蒸除溶剂并用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得4-(3-(3-(苄氧甲酰胺)-1-(异喹啉-6-甲酰胺)丙基)苯基)噻吩-2-羧酸乙酯(1.01g，1.68mmol，产率83％)。
MS(ESI)m/z＝594(M+1) +。
步骤8. 4-(3-(3-氨基-1-(异喹啉-6-甲酰胺)丙基)苯基)噻吩-2-羧酸乙酯的制备
Figure PCTCN2018091161-appb-000044
将4-(3-(3-(苄氧甲酰胺)-1-(异喹啉-6-甲酰胺)丙基)苯基)噻吩-2-羧酸乙酯(1.00g，1.68mmol)溶于醋酸(5.00mL)，室温下加入氢溴酸醋酸溶液(5.00ml)。室温搅拌3小时。减压蒸除溶剂。经MPLC纯化得4-(3-(3-氨基-1-(异喹啉-6-甲酰胺)丙基)苯基)噻吩-2-羧酸乙酯(50.3mg，0.09mmol，产率5.3％)。
MS(ESI)m/z＝594(M+1) +。
1H NMR(400MHz,MeOD):δ＝9.34(s,1H),8.55-8.54(m,2H),8.46(s,1H),8.25-8.22(m,1H),8.19-8.18(m,1H),8.12-8.09(m,1H),8.02-8.01(m,1H),7.97-7.96(m,1H),7.85(s,1H),7.70-7.67(m,1H),7.51-7.50(m,2H),5.40-5.37(m,1H),4.41-4.36(m,2H),3.36-3.12(m,1H),3.06-2.99(m,1H),2.51-2.31(m,2H),1.40(t,J＝6.8Hz,3H)。
实施例8、N-(3-(二甲氨基)-1-苯基-丙基)异喹啉-6-甲酰胺的制备
步骤1.叔丁基N-(3-(二甲氨基)-1-苯基-丙基)氨基酸甲酯的制备
Figure PCTCN2018091161-appb-000045
将叔丁基(1-(3-氯苯基)-2-氰乙基)氨基甲酸酯(500mg，1.78mmol)溶于甲醇(10.0 mL)，室温下加入钯碳(50.0mg)。在氢气下于室温搅拌3小时。加入甲醛溶液(535mg，17.8mmol)，在氢气下室温搅拌3小时。过滤除去固体，减压蒸除溶剂。柱层析纯化得叔丁基N-(3-(二甲氨基)-1-苯基-丙基)氨基酸甲酯(402mg，1.44mmol，产率81％)。
MS(ESI)m/z＝279(M+1) +。
步骤2.N',N'-二甲基-1苯基-丙基-1,3-二胺的制备
Figure PCTCN2018091161-appb-000046
将叔丁基N-(3-(二甲氨基)-1-苯基-丙基)氨基酸甲酯(200mg，0.72mmol)溶于甲醇(5.00mL)，室温下加入浓盐酸(5.00ml)。室温搅拌3小时，减压蒸除溶剂。经MPLC纯化得N',N'-二甲基-1苯基-丙基-1,3-二胺(121mg，0.67mol，产率94％)。
MS(ESI)m/z＝179(M+1) +。
步骤3.N-(3-(二甲氨基)-1-苯基-丙基)异喹啉-6-甲酰胺的制备
Figure PCTCN2018091161-appb-000047
将6-异喹啉甲酸(100mg，570μmol)溶于DMF(5.00mL)，室温下加入HBTU(219mg，570μmol)、DIEA(224mg，1.73mmol)和N',N'-二甲基-1苯基-丙基-1,3-二胺(100mg，570μmol)。室温搅拌3小时。减压蒸除溶剂并用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得N-(3-(二甲氨基)-1-苯基-丙基)异喹啉-6-甲酰胺(39.6mg，120μmol，产率21％)。
MS(ESI)m/z＝334(M+1) +。
1H NMR(400MHz,MeOD):δ＝9.80(s,1H),8.74(s,1H),8.46(s,1H),8.66(d,J＝6.0Hz,1H),8.58-8.54(m,2H),8.37-8.35(m,1H),7.56-7.53(m,1H),7.46-7.43(m,1H),7.38-7.34(m,1H),5.34-5.31(m,1H),3.48-3.36(m,1H),3.29-3.19(m,1H),2.96(s,6H),2.56-2.46(m,1H),2.44-2.35(m,1H).
实施例9、(N-氮杂环丁烷-基-苯甲基)异喹啉-6-甲酰胺的制备。
步骤1. 1-二苯甲基-N-甲氧基-N-甲基-氮杂环丁烷-3-甲酰胺的制备
Figure PCTCN2018091161-appb-000048
室温下往1-苯基氮杂环丁烷-3-甲酸(10.0g，37.4mmol)的二氯甲烷(100mL)溶液中加入N,N-二异丙基乙胺(19.3g，150mmol)、N-羟基-7-偶氮苯并三氮唑(5.56g，41.2mmol)和1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(7.90g，41.2mmol)后室温搅拌4小时，用乙酸乙酯萃取，再将水相用乙酸乙酯和水萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得到1-二苯甲基-N-甲氧基-N-甲基-氮杂环丁烷-3-甲酰胺(9.8g，29.0mmol，产率78％)。
MS(ESI)m/z＝311(M+1) +。
步骤2.(1-苯基氮杂环丁烷-3-基)-苯基-甲酮的制备
Figure PCTCN2018091161-appb-000049
将1-二苯甲基-N-甲氧基-N-甲基-氮杂环丁烷-3-甲酰胺(5.00g，16.1mmol)溶于四氢呋喃(50mL)，氮气保护下于0℃加入苯基溴化镁(80.6mL，1M，80.6mmol)，搅拌1小时后升至室温反应1小时。用饱和氯化铵溶液淬灭，乙酸乙酯萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得(1-苯基氮杂环丁烷-3-基)-苯基-甲酮(3.02g，8.25mmol，产率51％)。
MS(ESI)m/z＝328(M+1) +。
步骤3.(1-苄基氮杂环丁烷-3-基)-苯基-甲氨基的制备
Figure PCTCN2018091161-appb-000050
将(1-苯基氮杂环丁烷-3-基)-苯基-甲酮(500mg，1.53mmol)溶于甲醇(25mL)和二氯甲烷(25mL)，加入乙酸(90.0mg，1.53mmol)和乙酸铵(1.47g，6.11mmol)，氮气下反应1小时后，加入氰基硼氢化钠(380mg，6.11mmol)。反应液用饱和氯化铵溶液淬灭，乙酸乙酯萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得(1-苄基氮杂环丁烷-3-基)-苯基-甲氨基(199mg，0.53mmol，产率34％)。
MS(ESI)m/z＝329(M+1) +。
步骤4.四叔丁基-N-((1-苄基氮杂环丁烷-3-基)-苯基-甲基)氨基甲酸酯的制备
Figure PCTCN2018091161-appb-000051
将(1-苄基氮杂环丁烷-3-基)-苯基-甲氨基(1.20g，3.65mmol)溶于四氢呋喃(70mL)，加入碳酸钾(2.01g，14.6mmol)和二碳酸二叔丁酯(875mg，4.02mmol)，搅拌1小时后用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得四叔丁基-N-((1-苄基氮杂环丁烷-3-基)-苯基-甲基)氨基甲酸酯(1.10g，2.31mmol，产率63％)。
MS(ESI)m/z＝429(M+1) +。
步骤5.四叔丁基-N-(氮杂环丁烷-3-基(苯基)甲基)氨基甲酸酯的制备
Figure PCTCN2018091161-appb-000052
将四叔丁基-N-((1-苄基氮杂环丁烷-3-基)-苯基-甲基)氨基甲酸酯(1.20g，2.80mmol)溶于甲醇(10mL)，加入氢氧化钯(39mg，10％)后在氢气下搅拌24小时。用硅藻土过滤后，减压蒸除溶剂得四叔丁基-N-(氮杂环丁烷-3-基(苯基)甲基)氨基甲酸酯(503mg，1.52mmol，产率54％)。
MS(ESI)m/z＝263(M+1) +。
步骤6.烯丙基-3-((叔丁氧碳基酰胺)-苯基-甲基)氮杂环丁烷-1-甲酸酯的制备
Figure PCTCN2018091161-appb-000053
将四叔丁基-N-(氮杂环丁烷-3-基(苯基)甲基)氨基甲酸酯(1.00g，3.81mmol)溶于四氢呋喃(10mL)，加入三乙胺(770mg，7.62mmol)和氯甲酸烯丙酯(367g，3.05mmol)，搅拌2小时后用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得烯丙基-3-((叔丁氧碳基酰胺)-苯基-甲基)氮杂环丁烷-1-甲酸酯(597mg，1.39mmol，产率55％)。
MS(ESI)m/z＝347(M+1) +。
步骤7.烯丙基-3-(苯甲基胺)氮杂环丁烷-1-羧酸酯的制备
Figure PCTCN2018091161-appb-000054
将烯丙基-3-((叔丁氧碳基酰胺)-苯基-甲基)氮杂环丁烷-1-甲酸酯(500mg，1.44mmol)溶于甲醇(10.0mL)，冰浴下加入浓盐酸(3.00ml)，搅拌2小时后减压蒸除溶剂得烯丙基-3-(苯甲基胺)氮杂环丁烷-1-甲酸酯(121mg，0.39mol，产率27％)。
MS(ESI)m/z＝247(M+1) +。
步骤8.烯丙基-3-(异喹啉-6-甲酰胺)苯甲基)氮杂环丁烷-1-甲酸酯的制备
Figure PCTCN2018091161-appb-000055
室温下往烯丙基-3-(苯甲基胺)氮杂环丁烷-1-甲酸酯(150mg，610μmol)的DMF(10.0mL)溶液中加入N,N-二异丙基乙胺(315mg，2.44mmol)、N-羟基-7-偶氮苯并三氮唑(120mg，0.61mmol)和1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(156mg，0.61mmol)，室温搅拌4小时后用乙酸乙酯萃取，再将水相用乙酸乙酯和水萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得烯丙基-3-(异喹啉-6-甲酰胺)苯甲基)氮杂环丁烷-1-甲酸酯(121mg，0.27mmol，产率44％)。
MS(ESI)m/z＝402(M+1) +。
步骤9.N-(氮杂环丁烷-3-基-苯甲基)异喹啉-6-甲酰胺的制备
Figure PCTCN2018091161-appb-000056
室温下往烯丙基-3-(异喹啉-6-甲酰胺)苯甲基)氮杂环丁烷-1-甲酸酯(120mg，0.30mmol)的四氢呋喃(5.00mL)溶液中加入四三苯基膦钯(3.45mg，0.30mmol)和吗啉(260mg，2.99mmol)，室温搅拌2小时后减压蒸除溶剂。柱层析纯化得N-(氮杂环丁烷-3-基- 苯甲基)异喹啉-6-甲酰胺(6.0mg，0.14μmol，产率4.9％)。
MS(ESI)m/z＝318(M+1) +。
1H NMR(400MHz,DMSO-d 6+D 2O):δ＝9.78(s,1H),8.62-8.68(m,2H),8.49-8.55(m,2H),8.24-8.26(d,J＝8.8Hz,1H),7.31-7.45(m,5H),5.39-5.41(d,J＝10.4Hz,1H),4.12-4.16(m,1H),3.85-3.95(m,2H),3.72-3.78(m,1H),3.42-3.48(m,1H)。
实施例10、(甲基-N-氮杂环丁烷-苯基)异喹啉-6-甲酰胺的制备。
步骤1.四叔丁基(甲基-N-氮杂环丁烷-3-基(苯基)甲基)氨基甲酸酯的制备
Figure PCTCN2018091161-appb-000057
将四叔丁基-N-(氮杂环丁烷-3-基(苯基)甲基)氨基甲酸酯(1.00g，3.81mmol)溶于甲醇(10.0mL)，加入乙酸(266mg，3.81mmol)和甲醛水溶液(572mg，19.1mmol)，搅拌1小时后加入氰基硼氢化钠(239mg，3.81mmol)。用饱和氯化铵水溶液淬灭后，用乙酸乙酯萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得四叔丁基(甲基-N-氮杂环丁烷-3-基(苯基)甲基)氨基甲酸酯(796mg，2.03mmol，产率53％)。
MS(ESI)m/z＝277(M+1) +。
步骤2.(1-甲基氮杂环丁烷-3-基)-苯甲胺的制备
Figure PCTCN2018091161-appb-000058
将四叔丁基(甲基-N-氮杂环丁烷-3-基(苯基)甲基)氨基甲酸酯(773mg，2.80mmol)溶于甲醇(10mL)，冰浴下加入浓盐酸(3.00ml)，搅拌2小时后减压蒸除溶剂得烯丙基-3-(苯甲基胺)氮杂环丁烷-1-甲酸酯(0.48g，2.2mmol，产率78％)。
MS(ESI)m/z＝177(M+1) +。
步骤3.N-((1-甲基氮杂环丁烷-3-基)-苯甲基)异喹啉-6-甲酰胺的制备
Figure PCTCN2018091161-appb-000059
室温下往烯丙基-3-(苯甲基胺)氮杂环丁烷-1-甲酸酯(400mg，2.27mmol)的DMF(10.0 mL)溶液中加入N,N-二异丙基乙胺(1.17g，9.08mmol)、N-羟基-7-偶氮苯并三氮唑(430mg，2.27mmol)和1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(460mg，2.27mmol)，室温搅拌4小时，用乙酸乙酯萃取，再将水相用乙酸乙酯和水萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化到N-((1-甲基氮杂环丁烷-3-基)-苯甲基)异喹啉-6-甲酰胺(20.2mg，48.0μmol，产率2.1％)。
MS(ESI)m/z＝332(M+1) +。
1H NMR(400MHz,MeOD):δ＝9.86(s,1H),8.77(s,1H),8.60-8.68(m,3H),8.36-8.38(d,J＝8.8Hz,1H),7.52-7.54(d,J＝8.0Hz,2H),7.42-7.45(m,2H),7.36-7.38(m,1H),5.51-5.60(m,1H),4.44-4.58(m,1H),4.11-4.23(m,2H),3.96-4.02(m,1H),3.62-3.71(m,1H),2.96-3.01(d,J＝12Hz,3H)。
实施例11、N-((S)-苯基-((R)-四氢吡咯-3-基)甲基)异喹啉-6-甲酰胺和N-((R)-苯基-((R)-四氢吡咯-3-基)甲基)异喹啉-6-甲酰胺的制备
Figure PCTCN2018091161-appb-000060
步骤1.(R)-叔丁基3-(甲氧基(甲基)甲酰胺基)四氢吡咯-1-甲酸酯的制备
Figure PCTCN2018091161-appb-000061
将(R)-1-叔丁氧基羰基四氢吡咯-3-羧酸(15.0g，69.7mmol)溶于DCM(200mL)，加入EDCI(14.6g，76.7mmol)，HOBT(10.4g，76.7mmol)，DIEA(27.0g，209mmol)和二甲羟胺(6.38g，104.5mmol)，搅拌1小时后用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得叔丁基(R)-3-(甲氧基(甲基)氨基甲酰)四氢吡咯-1-甲酸酯(15g，58mmol，83％产率)。
MS(ESI)m/z＝259(M+1) +和203(M+1-56) +。
步骤2.叔丁基(R)-3-苯甲酰基四氢吡咯-1-甲酸酯的制备
Figure PCTCN2018091161-appb-000062
将叔丁基(R)-3-(甲氧基(甲基)氨基甲酰)四氢吡咯-1-甲酸酯(10g，38.7mmol)溶于THF(30mL)，苯基溴化镁(194mmol，1M，194mL)滴加入反应体系在0℃搅拌1小时 后用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得叔丁基(R)-3-苯甲酰基四氢吡咯-1-甲酸酯(9.1g，33mmol，84％产率)。
MS(ESI)m/z＝276(M+1) +和220(M+1-56) +。
步骤3.(R)-叔丁基-3-((羟亚胺)(苯基)甲基)四氢吡咯-1-甲酸酯的制备
Figure PCTCN2018091161-appb-000063
将叔丁基(R)-3-苯甲酰基四氢吡咯-1-甲酸酯(10g，36.3mmol)溶于MeOH(150mL)，室温下加入Na 2CO 3(11.6g，109mmol)和盐酸羟胺(6.00g，182mmol)，50℃搅拌2小时后用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得(R)-叔丁基-3-((羟亚胺)(苯基)甲基)四氢吡咯-1-甲酸酯(8.1g，28mmol，77％产率)。
MS(ESI)m/z＝291(M+1)和235(M+1-56) +。
步骤4.叔丁基(3R)-3-(氨基(苯基)甲基)四氢吡咯-1-甲酸酯的制备
Figure PCTCN2018091161-appb-000064
将(3R)-3-((E)-N-羟基-C-苯基-碳化二亚胺)四氢吡咯-1-甲酸酯(8.0g，27mmol)溶于MeOH(150.00mL)，室温下加入雷尼镍，氢气室温下搅拌2小时后用抽滤，滤液减压蒸除溶剂。柱层析纯化得叔丁基(R)-3-(氨基(苯基)甲基)四氢吡咯-1-甲酸酯(4.9g，18mmol，65％产率)。
MS(ESI)m/z＝277(M+1) +和221(M+1-56) +。
步骤5.叔丁基(R)-3-((异喹啉-6-甲酰胺)-苯基-甲基)四氢吡咯-1-甲酸酯的制备
Figure PCTCN2018091161-appb-000065
将异喹啉-6-甲酸(3.00g，17.3mmol)溶于DMF(20.0mL)，加入苯并三氮唑-N,N,N',N'-四甲基脲六氟磷酸酯(7.26g，19.06mmol)，叔丁基(R)-3-(氨基(苯基)甲基)四氢吡咯-1-甲酸酯(4.79g，17.3mmol)和N,N-二异丙基乙胺(3.38g，29.6mmol)，搅拌1小时后用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压 蒸除溶剂。柱层析纯化得叔丁基(R)-3-((异喹啉-6-甲酰胺)-苯基-甲基)四氢吡咯-1-甲酸酯(6.01g，13.9mmol，80％产率)。
MS(ESI)m/z＝432(M+1) +和376(M+1-56) +。
步骤6.N-((S)-苯基-((R)-四氢吡咯-3-基)甲基)异喹啉-6-甲酰胺和N-((R)-苯基-((R)-四氢吡咯-3-基)甲基)异喹啉-6-甲酰胺的制备
Figure PCTCN2018091161-appb-000066
将叔丁基(R)-3-((异喹啉-6-甲酰胺)-苯基-甲基)四氢吡咯-1-甲酸酯(5.00g，11.6mmol)溶于乙酸乙酯(30.0mL)，加入浓盐酸(10.0ml)，搅拌1小时后减压蒸除溶剂，经Pre-PLC制备得到N-((S)-苯基-((R)-四氢吡咯-3-基)甲基)异喹啉-6-甲酰胺(0.89g，2.69mmol，23％产率)和N-((R)-苯基-((R)-四氢吡咯-3-基)甲基)异喹啉-6-甲酰胺(1.0g，,3.0mmol，26％产率)。
MS(ESI)m/z＝332(M+1) +。
化合物11a: 1HNMR(400MHz,MeOD):δ＝9.31(s,1H),8.53-8.49(m,2H),8.39(s,1H),8.19(d,J＝8.8Hz,1H),8.04(d,J＝8.4Hz,1H),7.93(d,J＝5.6Hz,1H),7.55(d,J＝7.6Hz,2H),7.45-7.41(m,2H),7.37-7.33(m,1H),5.18(d,J＝10.0Hz,1H),3.55-3.49(m,1H),3.41-3.36(m,1H),3.19-3.11(m,2H),3.00-2.94(m,1H),2.45-2.39(m,1H),2.11-2.01(m,1H)。
化合物11b: 1HNMR(400MHz,MeOD):δ＝9.32(s,1H),8.53(d,J＝5.6Hz,1H),8.47(s,1H),8.39(s,1H),8.20(d,J＝8.4Hz,1H),8.05(d,J＝8.8Hz,1H),7.94(d,J＝6.0Hz,1H),7.54(d,J＝7.6Hz,2H),7.45-7.41(m,2H),7.37-7.33(m,1H),5.15(d,J＝10.8Hz,1H),3.69-3.64(m,1H),3.49-3.43(m,1H),3.31-3.26(m,2H),3.16-3.10(m,1H),1.93-1.85(m,1H),1.80-1.70(m,1H)。
实施例12、N-((R)-((R)-1-甲基四氢吡咯-3-基)(苯基)甲基)异喹啉-6-甲酰胺的制备
Figure PCTCN2018091161-appb-000067
将N-((R)-苯基-((R)-四氢吡咯-3-基)甲基)异喹啉-6-甲酰胺(250mg，1.51mmol)溶于甲醇(15.0mL)，加入乙酸(9.1mg，0.15mmol)和甲醛水溶液(0.58mL，13M，7.55mmol)，搅拌1小时后加入醋酸硼氢化钠(0.96g，4.53mmol)，继续反应2小时后用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。 柱层析纯化得N-((R)-((R)-1-甲基四氢吡咯-3-基)(苯基)甲基)异喹啉-6-甲酰胺(30mg，83umol，11％产率)。
MS(ESI)m/z＝346(M+1) +。
1HNMR(400MHz,DMSO-d 6):δ＝11.08-10.77(m,1H),9.77(s,1H),9.65-9.63(m,1H),8.76.-8.70(m,2H),8.47(d,J＝8.4Hz,1H),8.37(s 1H),8.27(t,J＝8.4Hz,1H),7.52(t,J＝8.0Hz,2H),7.41-7.36(m,2H),7.32-7.28(m,1H),5.16-5.07(m,1H),3.82-3.77(m,1H),3.55-3.48(m,2H),3.30-3.15(m,1H),3.13-3.05(m,1H),2.81(s,3H),1.79-1.61(m,2H)。
实施例13、N-((S)-((R)-1-甲基四氢吡咯-3-基)(苯基)甲基)异喹啉-6-甲酰胺的制备
Figure PCTCN2018091161-appb-000068
将N-((S)-苯基-((R)-四氢吡咯-3-基)甲基)异喹啉-6-甲酰胺(250mg，1.51mmol)溶于甲醇(15.0mL)，加入乙酸(9.1mg，0.15mmol)和甲醛水溶液(0.58mL，13M，7.55mmol)，搅拌1小时后加入醋酸硼氢化钠(0.96g，4.53mmol)，继续反应2小时后用乙酸乙酯和水萃取，再将水相用乙酸乙酯萃取两次，合并有机相并用无水硫酸钠干燥，减压蒸除溶剂。柱层析纯化得N-((S)-((R)-1-甲基四氢吡咯-3-基)(苯基)甲基)异喹啉-6-甲酰胺(21mg，58umol，7.7％产率)。
MS(ESI)m/z＝346(M+1) +。
1HNMR(400MHz,DMSO-d 6):δ＝11.17-10.90(m,1H),9.79(s,1H),9.65-9.48(m,1H),8.81.-8.70(m,2H),8.49(d,J＝8.4Hz,1H),8.40-8.39(m,1H),8.28(d,J＝8.4Hz,1H),7.57-7.49(m,2H),7.41-7.37(m,2H),7.32-7.29(m,1H),5.19-5.10(m,1H),3.63-3.51(m,2H),3.25-3.10(m,2H),3.08-2.91(m,1H),2.76-2.73(m,3H),2.37-2.23(m,1H),2.03-1.85(m,1H)。
实施例14、N-((S)-苯基-((S)-四氢吡咯-3-基)甲基)异喹啉-6-甲酰胺和N-((R)-苯基-((S)-四氢吡咯-3-基)甲基)异喹啉-6-甲酰胺的制备
Figure PCTCN2018091161-appb-000069
按照实施例11中步骤1到步骤6的方法，将步骤1中的(R)-1-叔丁氧基羰基四氢吡咯-3-羧酸替换为(S)-1-叔丁氧基羰基四氢吡咯-3-羧酸，得到N-((S)-苯基-((S)-四氢吡咯-3-基) 甲基)异喹啉-6-甲酰胺(28mg，总产率5.1％)和N-((R)-苯基-((S)-四氢吡咯-3-基)甲基)异喹啉-6-甲酰胺(22mg，总产率4.9％)。
MS(ESI)m/z＝332(M+1) +。
化合物14a: 1HNMR(400MHz,DMSO-d 6):9.96(s,1H),9.79-9.81(d,J＝8.4Hz,1H),9.51(s,2H),8.88(s,1H),8.75-8.76(d,J＝6Hz,1H),8.57-8.61(m,2H),8.37-8.40(m,1H),7.58-7.59(m,2H),7.36-7.40(m,2H),7.30-7.31(m,1H),5.09-5.14(m,1H),3.29-3.31(m,1H),3.36-3.18(m,1H),3.2-3.06(m,1H),2.83-2.89(m,2H),2.23-2.27(m,1H),1.82-1.88(m,1H)。
化合物14b: 1HNMR(400MHz,DMSO-d 6):δ＝9.90(s,1H),9.84(d,J＝7.6Hz,1H),9.64(s,H),9.50(s,H),8.88(s,1H),8.74(d,J＝6.4Hz,1H),8.49-8.57(m,2H),8.35(d,J＝8.8Hz,1H),7.55-7.57(m,2H),7.37-7.40(m,2H),7.30-7.31(m,1H),5.06(t,J＝6.0Hz,1H),3.52(m,1H),3.28(m,1H),3.06(m,3H),1.64(m,1H)。
实施例15、N-((S)-2-(二甲氨基)-1-(3-甲氧基苯基)乙基)异喹啉-6-甲酰胺的制备
步骤1.N-((1S)-2-(二甲氨基)-1-(3-甲氧基苯基)乙基)异喹啉-6-甲酰胺的制备
Figure PCTCN2018091161-appb-000070
按照实施例1中步骤1至步骤3、实施例5中步骤1至步骤3的方法，用3-甲氧基苯甲醛替代实施例1中步骤1中的苯甲醛，得到N-((S)-2-(二甲氨基)-1-(3-甲氧基苯基)乙基)异喹啉-6-甲酰胺。
MS(ESI)m/z＝350(M+1) +。
1HNMR(400MHz,MeOD):δ＝9.75(s,1H),8.87(s,1H),8.66(d,J＝6.4Hz,1H),8.55-8.53(m,1H),8.48(d,J＝6.4Hz,1H),8.45-8.42(m,1H),7.39(t,J＝8.0Hz,1H),7.20-7.16(m,2H),6.99-6.96(m,1H),5.80-5.76(m,1H),3.99-3.93(m,1H),3.85(s,1H),3.67-3.62(m,1H),3.11-3.08(m,6H)。
实施例16、N-((S)-2-(二甲氨基)-1-(3-氯苯基)乙基)异喹啉-6-甲酰胺的制备
步骤1.N-((S)-2-(二甲氨基)-1-(3-氯苯基)乙基)异喹啉-6-甲酰胺的制备
Figure PCTCN2018091161-appb-000071
按照实施例1中步骤1至步骤3、实施例5中步骤1至步骤3的方法，用3-氯苯甲醛替代实施例1中步骤1中的苯甲醛，得到N-((S)-2-(二甲氨基)-1-(3-甲氧基苯基)乙基)异喹 啉-6-甲酰胺。
MS(ESI)m/z＝354(M+1) +。
1HNMR(400MHz,MeOH):δ＝9.91(s,1H),9.06(s,1H),8.72-8.64(m,3H),8.57(d,J＝8.8Hz,2H),7.69(s,1H),7.48-7.41(m,2H),5.82(d,J＝11.6Hz,1H),4.06(t,J＝11.6Hz,1H),3.64(t,J＝13.6Hz,1H),3.12(s,3H),3.08(s,3H)。
为了说明本发明的有益效果，本发明提供以下试验例：
试验例1、ROCK2抑制活性的检测
ROCK2能够磷酸化S6K(KRRRLASLR)多肽底物，将ATP转化成ADP。在激酶反应后，加入ADP-Glo TM试剂，使激酶反应终止，并消耗完剩余的ATP。加入激酶检测试剂，它在使ADP转化成ATP的同时，ATP再被Ultra-Glo TM萤光素酶转化成光发光信号，发光信号与激酶活性正相关。
按以下步骤进行ROCK2抑制活性的检测：
1.检测缓冲液:40mM Tris pH 7.5，20mM MgCl 2，0.1％BSA(w/v)，50μM DTT；
2.加12μL2.5x0.1μg/ml ROCK2工作液进入96孔PCR板；
3.加6μL6x化合物工作液进入96孔PCR板混匀，25℃预孵育10min；
4.加入12μL 2.5x 37.5μg/ml S6K底物和12.5μM ATP混合工作液，30℃孵育60min；
5.取25μL反应混合物到一个新96孔PCR板，并加入25μL ADP-Glo TM试剂混匀，25℃孵育40min终止反应；
6.取40μL终止反应混合物到一个新96孔PCR板，并加入40μL激酶检测试剂混匀，25℃孵育40min；
7.读取luminescence(冷光)信号值，计算抑制率。
按照上述方法对实施例制备的化合物进行ROCK2抑制活性检测，试验结果见表1，其中测定各化合物的IC 50按照说明分类，表1中：
“+”表示IC 50测定值大于500nM；
“++”表示IC 50测定值小于500nM大于100nM；
“+++”表示IC 50测定值小于100nM
表1、化合物对ROCK2的抑制活性
实施例化合物 ROCK2 实施例化合物 ROCK2
1 +++ 2 +++
3 ++ 4 ++
5 +++ 6 +++
7 +++ 8 +++
9 +++ 10 +++
11a +++ 11b +++
12 +++ 13 +++
14a ++ 14b +++
15 +++    
上述实验表明，本发明的化合物对ROCK2具有良好的酶学抑制活性。
试验例2、细胞原位免疫印迹(In-Cell Western)检测肌球蛋白轻链磷酸化
使用大鼠平滑肌细胞系A7r5。ROCK2通过磷酸化肌球蛋白轻链T18/S19两个氨基酸位点导致细胞骨架的改变。将A7r5细胞接种入96孔透明底黑色板，培养条件为含10％FBS的DMEM。过夜培养后，血清饥饿4小时，在无血清培养基中将细胞与化合物一起温育1小时。
使用phspho-MLC-T18/S19特异性抗体和第二检测抗体，通过细胞原位免疫印迹检测肌球蛋白轻链磷酸化水平。未处理的细胞作为阳性对照，一抗未孵育样品作为阴性对照，细胞内总蛋白作为内参。使用GraphPad Prism 5.01软件，使用具有变化斜率的非线性回归曲线拟合，确定EC 50值。
按照上述方法对实施例制备的化合物进行大鼠平滑肌细胞系A7r5肌球蛋白轻链磷酸化抑制活性检测，试验结果见表1，其中测定各化合物的IC 50按照说明分类，表1中：
“+”表示IC 50测定值大于1μM；
“++”表示IC 50测定值小于1μM大于250nM；
“+++”表示IC 50测定值小于250nM
表2、化合物对大鼠平滑肌细胞系A7r5的抑制活性
实施例化合物 A7r5 实施例化合物 A7r5
1 ++ 2 +++
4 + 5 ++
6 +++ 7 ++
8 + 9 ++
10 ++ 11a ++
13 ++ 15 ++
16 ++    
上述试验结果表明，本发明的化合物对大鼠平滑肌细胞系A7r5具有良好的抑制活性，进而，本发明的化合物可以抑制由ROCK2通过磷酸化肌球蛋白轻链T18/S19两个氨基酸位点导致细胞骨架的改变，并具有治疗与此相关疾病的潜在活性。
试验例3、动物实验(正常眼压模型)
将24只雄性新西兰白兔分为4组，每组6只。使用考虑了体重的计算机随机化算法，将动物随机分配到各组当中。在实验开始前3天至前1天，所有动物需要被训练适应眼压测量的操作过程，每天应进行3次，间隔约3小时。眼压测定使用TonoVet眼压计，固定测试动物，并将待测眼置于TonoVet眼压计相垂直的位置上。测量至少3次眼压，直至眼压读数稳定。取最后3次稳定的测量数据(±3mmHg)，取均值，视作最终眼压值。称取检测化合物，用生理盐水溶解成使用浓度。实验开始时，待测化合物、对照品K115、或阴性对照溶剂(50μL)将被施于每只动物的右眼，左眼则滴加生理盐水。在滴加药物之后的第Pre,1,2,4,6,8,10小时之后测量眼压。使用student-t检验来分析单个时间点的数据。眼压随时间的变化曲线将使用ANOVA检验来分析。P值小于0.05将被视为具有统计学显著性，数据的展示方式是平均值正负标准误。
实验结果(见图1、图2、图3)表明，本发明化合物5与化合物11a具有与对照品K115类似的降眼压活性，并且具有更长的作用时间。
试验例4、动物实验(高眼压单眼造模模型)
将10只雄性新西兰白兔分为2组，每组5只。使用考虑了体重的计算机随机化算法，将动物随机分配到各组当中。在实验开始前3天至前1天，所有动物需要被训练适应眼压测量的操作过程，每天应进行3次，间隔约3小时。眼压测定使用TonoVet眼压计，固定测试动物，并将待测眼置于TonoVet眼压计相垂直的位置上。测量至少3次眼压，直至眼压读数稳定。取最后3次稳定的测量数据(±3mmHg)，取均值，视作最终眼压值。称 取检测化合物，用生理盐水溶解成使用浓度。诱导高眼压前，测量双眼的眼压值，作为基础眼压记录。随后，通过静脉注射(50％)戊巴比妥钠50mg/kg对白兔进行麻醉。使用尖30-G针建立临时前房穿刺，向白兔右眼前房注射50μL的粘性物质，以诱导高眼压，对侧眼给予相同体积的生理盐水。随后，使用棉签按压前房穿刺，以防止房水回流。每组造模动物的右眼給予受试化合物，一天三次分别在第0,3,6小时给药1次,50μL/眼。对测眼将在相同时间点給予相同体积的生理盐水或特定溶媒。第0,1,2,3,4,6,8小时之后测量眼压。使用student-t检验来分析单个时间点的数据。眼压随时间的变化曲线将使用ANOVA检验来分析。P值小于0.05将被视为具有统计学显著性，数据的展示方式是平均值正负标准误。
实验结果(见图4)表明，本发明的化合物11a对高眼压具有良好的降压活性。
本发明的上述试验表明，本发明实施例的化合物具有良好的ROCK抑制活性，可以有效用于与ROCK活性异常疾病的治疗。
综上所述，本发明公开的式I所示的新化合物，表现出了良好的ROCK抑制活性，为临床治疗与ROCK活性异常相关的疾病提供了一种新的药用可能。

Claims (12)

  1. 式(Ⅰ)所示的化合物或其立体异构体：
    Figure PCTCN2018091161-appb-100001
    其中，
    n分别为0、1或2；
    R 1分别独立地选自氢、羟基、卤素、氨基、羧基、三氟甲基、硝基、氰基、C 1～6烷基；
    m分别为0、1、2、3、4或5；
    R 3选自-NR 2R 2’、含N的杂环烷基；其中，含N的杂环烷基可以进一步被1～2个独立选自于卤素、C 1～6烷基的取代基取代；
    R 2、R 2’分别独立选自氢、C 1～6烷基；
    A环选自5～6元芳环、5～6元芳杂环或
    Figure PCTCN2018091161-appb-100002
    B环、C环分别独自选自5～6元芳环、5～6元芳杂环；
    R 4分别独立地选自氢、卤素、硝基、氰基、C 1～6烷基、C 2～6烯基、C 2～6炔基、3～6元环烷基、3～6元杂环烷基、-(CH 2) mOR a、-(CH 2) mOC(O)R a、-(CH 2) mOC(O)NR aR b、-(CH 2) mNR aR b、-(CH 2) mNR aC(O)R b、-(CH 2) mNR aC(O)OR b、-(CH 2) mC(O)R a、-(CH 2) mC(O)OR a、-(CH 2) mC(O)NR aR b；
    R a、R b分别独自选自氢、C 1～6烷基、C 2～6烯基、C 2～6炔基、3～6元环烷基、3～6元杂环烷基、5～6元芳环、5～6元芳杂环；其中，3～6元环烷基、3～6元杂环烷基、5～6元芳环、5～6元芳杂环可以进一步被1～2个独立选自于卤素、C 1～6烷基的取代基取代。
  2. 根据权利要求1所述的化合物或其立体异构体，其特征是：所述R 3选自的含N的杂环烷基为4～5元的含N杂环烷基。
  3. 根据权利要求1所述的化合物或其立体异构体，其特征是：当R 3选自含N的杂环烷基且A环有2个R 4取代时，2个R 4不同时为卤素。
  4. 根据权利要求1所述的化合物或其立体异构体，其特征是：所述化合物的结构如式(Ⅱ)所示：
    Figure PCTCN2018091161-appb-100003
    其中，
    R 1选自氢、羟基、卤素、氨基、羧基、三氟甲基、硝基、氰基、C 1～6烷基；
    m分别为0、1、2或3；
    R 2、R 2’分别独立选自氢、C 1～6烷基；
    A环选自5～6元芳环、5～6元芳杂环或
    Figure PCTCN2018091161-appb-100004
    B环、C环分别独自选自5～6元芳环、5～6元芳杂环；
    R 4分别独立地选自氢、卤素、硝基、氰基、C 1～6烷基、C 2～6烯基、C 2～6炔基、3～6元环烷基、3～6元杂环烷基、-(CH 2) mOR a、-(CH 2) mOC(O)R a、-(CH 2) mNR aR b、-(CH 2) mNR aC(O)R b、-(CH 2) mC(O)R a、-(CH 2) mC(O)OR a、-(CH 2) mC(O)NR aR b；
    R a、R b分别独自选自氢、C 1～6烷基、C 2～6烯基、C 2～6炔基、3～6元环烷基、3～6元杂环烷基、5～6元芳环、5～6元芳杂环；其中，3～6元环烷基、3～6元杂环烷基、5～6元芳环、5～6元芳杂环可以进一步被1～2个独立选自于卤素、C 1～6烷基的取代基取代。
  5. 根据权利要求4所述的化合物或其立体异构体，其特征是：所述化合物如下列结构式所示：
    Figure PCTCN2018091161-appb-100005
    Figure PCTCN2018091161-appb-100006
  6. 根据权利要求1所述的化合物或其立体异构体，其特征是：所述化合物的结构如式(Ⅲ)所示：
    Figure PCTCN2018091161-appb-100007
    R 1选自氢、羟基、卤素、氨基、羧基、三氟甲基、硝基、氰基、C 1～6烷基；
    m分别为0、1、2或3；
    A环选自5～6元芳环、5～6元芳杂环或
    Figure PCTCN2018091161-appb-100008
    B环、C环分别独自选自5～6元芳环、5～6元芳杂环；
    E环选自含N的杂环烷基；所述含N的杂环烷基可以进一步被1～2个独立选自于卤素、C 1～6烷基的取代基取代；
    R 4分别独立地选自氢、卤素、硝基、氰基、C 1～6烷基、C 2～6烯基、C 2～6炔基、3～6元环烷基、3～6元杂环烷基、-(CH 2) mOR a、-(CH 2) mOC(O)R a、-(CH 2) mNR aR b、-(CH 2) mNR aC(O)R b、-(CH 2) mC(O)R a、-(CH 2) mC(O)OR a、-(CH 2) mC(O)NR aR b；
    R a、R b分别独自选自氢、C 1～6烷基、C 2～6烯基、C 2～6炔基、3～6元环烷基、3～6元杂环烷基、5～6元芳环、5～6元芳杂环；其中，3～6元环烷基、3～6元杂环烷基、5～6元芳环、5～6元芳杂环可以进一步被1～2个独立选自于卤素、C 1～6烷基的取代基取代。
  7. 根据权利要求6所述的化合物或其立体异构体，其特征是：E环选自4～5元的含N的杂环烷基。
  8. 根据权利要求7所述的化合物或其立体异构体，其特征是：所述化合物如下列结构式所示：
    Figure PCTCN2018091161-appb-100009
  9. 权利要求1～8任意一项所述化合物或其立体异构体、晶型、药学上可接受的盐、水合物或溶剂合物在制备治疗与ROCK活性异常相关的疾病的药物中的用途。
  10. 根据权利要求9所述用途，其特征在于：所述与ROCK活性异常相关的疾病是细胞骨架调整、平滑肌细胞收缩、神经再生相关的疾病。
  11. 根据权利要求10所述用途，其特征在于：所述的疾病为高眼压症或者青光眼。
  12. 一种药物，其特征在于：它是以权利要求1～8任一项所述的化合物或其立体异构体、晶型、药学上可接受的盐、水合物或溶剂合物为活性成分，加上药学上可接受的辅料制备而成的制剂。
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CN115052855A (zh) * 2020-03-06 2022-09-13 昊运股份有限公司 酰胺化合物的制备方法、其晶型及其盐类

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101253152A (zh) * 2005-09-02 2008-08-27 安斯泰来制药株式会社 作为rock抑制剂的酰胺衍生物
CN101321748A (zh) * 2005-12-08 2008-12-10 欧加农股份有限公司 异喹啉衍生物
CN102131784A (zh) * 2008-06-24 2011-07-20 赛诺菲-安万特 作为rho激酶抑制剂的取代的异喹啉和异喹啉酮

Family Cites Families (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1716137A1 (en) * 2004-02-04 2006-11-02 Pfizer Products Incorporated Substituted quinoline compounds
DE602008003055D1 (de) * 2007-05-31 2010-12-02 Nerviano Medical Sciences Srl Zyklokondensierte Azaindole, die als Kinase-Hemmer wirken
US8648069B2 (en) * 2007-06-08 2014-02-11 Abbvie Inc. 5-substituted indazoles as kinase inhibitors
WO2010056758A1 (en) * 2008-11-12 2010-05-20 Yangbo Feng Quinazoline derivatives as kinase inhibitors
US8394826B2 (en) * 2009-05-01 2013-03-12 Aerie Pharmaceuticals, Inc. Dual mechanism inhibitors for the treatment of disease
JP2014076948A (ja) * 2011-02-09 2014-05-01 Astellas Pharma Inc イソキノリンアミド誘導体
WO2014036015A1 (en) * 2012-08-27 2014-03-06 Array Biopharma Inc. Serine/threonine kinase inhibitors for the treatment of hyperproliferative|diseases
ES2633987T3 (es) * 2013-07-02 2017-09-26 Bristol-Myers Squibb Company Derivados de pirido-carboxamidas tricíclicas como inhibidores de ROCK
CN109134433B (zh) * 2017-06-16 2021-05-25 成都先导药物开发股份有限公司 一种抑制rock的化合物及其应用
WO2019201296A1 (zh) * 2018-04-18 2019-10-24 南京明德新药研发有限公司 作为rho激酶抑制剂的吡唑类化合物
WO2020172615A1 (en) * 2019-02-22 2020-08-27 Glaukos Corporation Compounds for the treatment of ocular disease

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101253152A (zh) * 2005-09-02 2008-08-27 安斯泰来制药株式会社 作为rock抑制剂的酰胺衍生物
CN101321748A (zh) * 2005-12-08 2008-12-10 欧加农股份有限公司 异喹啉衍生物
CN102131784A (zh) * 2008-06-24 2011-07-20 赛诺菲-安万特 作为rho激酶抑制剂的取代的异喹啉和异喹啉酮

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
"Acute vasodilator effects of a Rho-kinase inhibitor, fasudil, in patients with severe pulmonary hypertension [J", HEART, vol. 91, no. 3, 2005, pages 391 - 2
"Involvement of Rho-kinase in hypertensive vascular disease: a novel therapeutic target in hypertension [J].", FASEB J., vol. 15, no. 6, 2001, pages 1062 - 4
See also references of EP3640242A4
TAKAMI AIWAKUBO MOKADA Y ET AL.: "Design and synthesis of Rho kinase inhibitors [J", BIOORG MED CHEM, vol. 12, 2004, pages 2115 - 2137, XP002309639, DOI: 10.1016/j.bmc.2004.02.025
TAMURA MNAKAO HYOSHIZAKI H ET AL.: "Development of specific Rho-kinase inhibitors and their clinical application [J", BIOCHIM BIOPHYS ACTA, vol. 1754, 2005, pages 245 - 252, XP005214213, DOI: 10.1016/j.bbapap.2005.06.015
UEHATA MISHIZAKI TSATOH H ET AL.: "Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension [J", NATURE, vol. 389, no. 6654, 1997, pages 990 - 994, XP002229794, DOI: 10.1038/40187
YARROW JCTOTSUKAWA GCHARRAS GT ET AL.: "Screening for cell migration inhibitors via automated microscopy reveals a Rho-kinase inhibitor [J", CHEM BIOL, vol. 12, 2005, pages 385 - 395, XP004818907, DOI: 10.1016/j.chembiol.2005.01.015

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