WO2019201296A1 - Composé de pyrazole tenant lieu d'inhibiteur de la rho-kinase - Google Patents

Composé de pyrazole tenant lieu d'inhibiteur de la rho-kinase Download PDF

Info

Publication number
WO2019201296A1
WO2019201296A1 PCT/CN2019/083208 CN2019083208W WO2019201296A1 WO 2019201296 A1 WO2019201296 A1 WO 2019201296A1 CN 2019083208 W CN2019083208 W CN 2019083208W WO 2019201296 A1 WO2019201296 A1 WO 2019201296A1
Authority
WO
WIPO (PCT)
Prior art keywords
group
compound
och
isomer
pharmaceutically acceptable
Prior art date
Application number
PCT/CN2019/083208
Other languages
English (en)
Chinese (zh)
Inventor
吴凌云
尹军
王才林
肖哲明
黎健
陈曙辉
Original Assignee
南京明德新药研发有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 南京明德新药研发有限公司 filed Critical 南京明德新药研发有限公司
Publication of WO2019201296A1 publication Critical patent/WO2019201296A1/fr

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/415—1,2-Diazoles
    • 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
    • A61K31/4709—Non-condensed quinolines and containing further heterocyclic rings
    • 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
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
    • C07D231/02—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings
    • C07D231/10—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
    • C07D231/12—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
    • 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
    • C07D401/04—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 directly linked by a ring-member-to-ring-member bond

Definitions

  • the present invention relates to the field of medicine, and in particular to a class of pyrazole compounds as RHO kinase inhibitors, pharmaceutically acceptable salts thereof or isomers thereof, pharmaceutical compositions thereof and their use in the preparation of RHO inhibitor drugs.
  • Rho associated kinase a serine/threonine protein kinase
  • RHO-associated protein kinase is involved in the regulation of myosin light chain (MLC) and is suitable for the treatment of vasodilation.
  • MLC myosin light chain
  • New research supports the regulation of ROCK kinase involved in the immune response of TH17 cells and the activation of fibroblasts, which can be expanded and adapted.
  • the disease is used for lung fibrosis, asthma and other lung diseases, and further indications include autoimmune diseases.
  • the ROCK kinase family includes two subtypes, ROCK1 and ROCK2.
  • ROCK2 kinase is involved in inflammation and fibrosis. Selective ROCK2 inhibitors do not cause vasodilation at high concentrations in ex vivo vasodilation and reduce cardiovascular side effects. Although the embryonic mortality rate of ROCK1 knockout mice is not high, most of them die after cytoskeletal variation caused by decreased MLC phosphorylation, while 90% of ROCK2 knockout mice die during embryonic stage, but surviving mice and wild type Without distinction, selective inhibition of ROCK2 activity may be more secure. Therefore, selective ROCK2 protein kinase inhibitors can avoid the cardiovascular side effects of drugs.
  • KD025 (WO2006105081; WO2008054599; WO2010104851; WO2014055996) is an oral ROCK2 kinase selective inhibitor developed by Kadmon Corporation. Studies have shown that KD025 is a new mechanism for the treatment of idiopathic pulmonary fibrosis (IPF) by inhibiting proteins such as RHO kinase that regulate fibrosis. The cause of idiopathic pulmonary fibrosis (IPF) may be body damage.
  • IPF idiopathic pulmonary fibrosis
  • the body's response to injury involves the reorganization of the actin cytoskeleton of a variety of cells (such as epithelial cells, fibroblasts, endothelial cells, and macrophages, etc.), and the assembly of actin filaments and myospheres.
  • the contraction of the protein (actomyosin) is regulated by RHO kinase family proteins, including ROCK1 and ROCK2.
  • RHO kinase family proteins include ROCK1 and ROCK2.
  • Previous studies have shown that RHO kinase family proteins are activated in the lungs of IPF patients and animal models of the disease, while RHO kinase inhibitors can prevent tissue fibrosis in these models and induce regression of established fibrosis. .
  • Phase II clinical trials for the treatment of moderate to severe psoriasis have been completed and are in Phase II clinical studies at the stage of idiopathic pulmonary fibrosis (IPF).
  • IPPF idiopathic pulmonary
  • WO2014134388 and WO2016028971 also disclose a class of compounds having the general formula shown in formula (a) and formula (b). Such compounds are also useful in cardiovascular diseases, neuropathological diseases, tumors, autoimmune diseases, pulmonary fibrosis. , treatment of inflammatory diseases, etc.
  • the compound of the present invention is a ROCK2 inhibitor with a certain selectivity for ROCK2, which has remarkable kinase inhibitory activity, membrane permeability and solubility, and has excellent pharmacokinetic and pharmacodynamic properties.
  • the present invention provides a compound of the formula (I-1), a pharmaceutically acceptable salt thereof or an isomer thereof,
  • said C 1-6 alkyl group and C 3-6 cycloalkyl group are optionally 1, 2 or 3 independently selected from the group consisting of F, Cl, Br, I, -OH, -OCH 3 , -CN, -NH 2, -NO 2 or C 1-4 alkyl substituents;
  • T 1 and T 2 are each independently N or CH;
  • R 6 and R 7 are each independently H, F, Cl or C 1-6 alkyl, wherein said C 1-6 alkyl group is optionally 1, 2 or 3 independently selected from the group consisting of F, Cl, Br, CN Substituted by a substituent of -OR a or -NR d R e ;
  • Each R 8 is independently F, Cl, Br, CN, -OR a , C 1-6 alkyl or C 3-6 cycloalkyl, wherein said C 1-6 alkyl and C 3-6 cycloalkyl Optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of F, Cl, Br, I, -OH, -OCH 3 , -CN, -NH 2 , -NO 2 or C 1-4 alkyl;
  • R a , R b and R c are each independently H or C 1-6 alkyl, wherein the C 1-6 alkyl group is optionally 1, 2 or 3 independently selected from the group consisting of F, Cl, Br, -OH Substituted with a substituent of -OCH 3 , -CN, -NH 2 , C 1-6 alkylamino or -NO 2 ;
  • R d and R e are each independently H, C 1-6 alkyl, or R d and R e together with the N atom to which they are attached form a 4-8 membered heterocycloalkyl group, wherein said C 1-6 alkyl group And a 4-8 membered heterocycloalkyl group is optionally 1, 2 or 3 independently selected from the group consisting of F, Cl, Br, -OH, -OCH 3 , -CN, -NH 2 , C 1-6 alkylamino or -NO 2 substituents;
  • n 0, 1, 2, 3 or 4;
  • the 4-8 membered heterocycloalkyl contains 1, 2, 3 or 4 heteroatoms or heteroatoms independently selected from -O-, -S-, N or -NH-.
  • R a , R b and R c are each independently H, methyl, ethyl, n-propyl or isopropyl, wherein the methyl, ethyl, n-propyl and The isopropyl group is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of F, Cl, Br, I, -OH, -OCH 3 , CN, -NH 2 , C 1-3 alkylamino or -NO 2
  • substituents independently selected from the group consisting of F, Cl, Br, I, -OH, -OCH 3 , CN, -NH 2 , C 1-3 alkylamino or -NO 2
  • other variables are as defined by the present invention.
  • R a , R b and R c are each independently H, methyl, ethyl, n-propyl, isopropyl, Other variables are as defined by the present invention.
  • the cyclohexyl group is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of F, Cl, Br, I, -OH, -OCH 3 , CN, -NH 2 , -NO 2 , methyl, ethyl or propyl.
  • other variables are as defined by the present invention.
  • R 3 , R 4 and R 5 and other variables are as defined by the present invention.
  • R d and R e are each independently H, methyl, ethyl, n-propyl, isopropyl, or R d and R e are taken together with the N atom to which they are attached 5- a 6-membered heterocycloalkyl group, wherein the methyl, ethyl, n-propyl, isopropyl and 5-6 membered heterocycloalkyl are optionally 1, 2 or 3 independently selected from the group consisting of F, Cl, Br, I, -OH, -OCH 3, CN , -NH 2, C 1-3 alkylamino or a -NO 2 substituents other variables are as defined in the present invention.
  • R d and R e are each independently H, methyl, ethyl, n-propyl, isopropyl, Or R d and R e together with the N atom to which they are attached form a pyrrolidinyl or piperidinyl group, wherein the pyrrolidinyl and piperidinyl are optionally 1, 2 or 3 independently selected from the group consisting of F, Cl, Br, I, -OH, -OCH 3, CN , -NH 2, C 1-3 alkylamino or a -NO 2 substituents other variables are as defined in the present invention.
  • R 6 and R 7 are each independently H, F, Cl, -OH, -OCH 3 , -NH 2 , Methyl, ethyl, n-propyl, isopropyl, Other variables are as defined by the present invention.
  • R 6 and R 7 are each independently H, F, Cl, methyl, ethyl, n-propyl, isopropyl, Other variables are as defined by the present invention.
  • each of R 8 above is independently F, Cl, Br, CN, -OH, -OCH 3 , -OCH 2 F, -OCHF 2 , -OCF 3 , methyl, ethyl, ortho a propyl group, an isopropyl group, a cyclopropyl group, a cyclopentyl group or a cyclohexyl group, wherein the methyl group, ethyl group, n-propyl group, isopropyl group, cyclopropyl group, cyclopentyl group and cyclohexyl group are optionally 1, 2 or 3 substituents independently selected from the group consisting of F, Cl, Br, I, -OH, -OCH 3 , CN, -NH 2 , -NO 2 , methyl, ethyl or propyl, other variables such as The invention is defined.
  • each of R 8 above is independently F, Cl, Br, CN, -OH, -OCH 3 , -OCH 2 CH 3 , methyl, ethyl, n-propyl, isopropyl, Cyclopropyl, cyclopentyl, cyclohexyl, Other variables are as defined by the present invention.
  • the above compound, a pharmaceutically acceptable salt thereof or an isomer thereof has a structure represented by the formula (I-2) to (I-4),
  • R 1 , R 2 , R 3 , R 6 , R 7 , R 8 and n are as defined in the present invention.
  • the above compound, a pharmaceutically acceptable salt thereof or an isomer thereof has a structure represented by formula (I-5) to (I-7),
  • R 3 , R 6 , R 7 , R 8 and n are as defined in the present invention.
  • the above compound, a pharmaceutically acceptable salt thereof or an isomer thereof has a structure represented by the formula (I-8) to (I-10),
  • carbon atom bearing "*" is a chiral carbon atom, which is in the form of a single enantiomer of (R) or (S) or is enriched in one enantiomer form;
  • R 7 is F, Cl, -OH, -OCH 3 , -NH 2 , Methyl, ethyl, n-propyl, isopropyl, R 3 , R 8 and n are as defined in the present invention.
  • the above compound, a pharmaceutically acceptable salt thereof or an isomer thereof has a structure represented by the formula (I-8) to (I-10),
  • carbon atom bearing "*" is a chiral carbon atom, which is in the form of a single enantiomer of (R) or (S) or is enriched in one enantiomer form;
  • R 7 is F, Cl, methyl, ethyl, n-propyl, isopropyl, R 3 , R 8 and n are as defined in the present invention.
  • the above compound is selected from the group consisting of a compound of the formula: a pharmaceutically acceptable salt thereof or an isomer thereof:
  • the pharmaceutically acceptable salt of the above compound is a formate or hydrochloride salt.
  • the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising, as an active ingredient, a therapeutically effective amount of the above compound, a pharmaceutically acceptable salt thereof or an isomer thereof, and a pharmaceutically acceptable carrier.
  • the present invention provides the use of the above compound, a pharmaceutically acceptable salt thereof or an isomer thereof, and the above pharmaceutical composition for the preparation of a medicament for RHO inhibitor.
  • the present invention also provides the above compound, a pharmaceutically acceptable salt thereof or an isomer thereof, and the use of the above pharmaceutical composition for the preparation of a medicament for treating an autoimmune disease; in some embodiments, the autoimmune disease is I Type 2 diabetes, rheumatoid arthritis, inflammatory bowel disease, systemic lupus erythematosus, glaucoma, pulmonary fibrosis, psoriasis or multiple sclerosis.
  • the autoimmune disease is I Type 2 diabetes, rheumatoid arthritis, inflammatory bowel disease, systemic lupus erythematosus, glaucoma, pulmonary fibrosis, psoriasis or multiple sclerosis.
  • the compound of the present invention has remarkable kinase inhibitory activity against ROCK2, and has certain selectivity to ROCK2, and has certain membrane permeability and solubility, and has excellent pharmacokinetic and pharmacodynamic properties.
  • pharmaceutically acceptable as used herein is intended to mean that those compounds, materials, compositions and/or dosage forms are within the scope of sound medical judgment and are suitable for use in contact with human and animal tissues. Without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit/risk ratio.
  • pharmaceutically acceptable salt refers to a salt of a compound of the invention prepared from a compound having a particular substituent found in the present invention and a relatively non-toxic acid or base.
  • a base addition salt can be obtained by contacting a neutral amount of such a compound with a sufficient amount of a base in a neat solution or a suitable inert solvent.
  • Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts.
  • an acid addition salt can be obtained by contacting a neutral form of such a compound with a sufficient amount of an acid in a neat solution or a suitable inert solvent.
  • pharmaceutically acceptable acid addition salts include inorganic acid salts including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, hydrogencarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, Hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and an organic acid salt, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, Similar acids such as fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and me
  • the pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing an acid group or a base by conventional chemical methods.
  • such salts are prepared by reacting these compounds in water or an organic solvent or a mixture of the two via a free acid or base form with a stoichiometric amount of a suitable base or acid.
  • the compounds of the invention may exist in specific geometric or stereoisomeric forms. All such compounds are contemplated by the present invention, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereoisomers , (D)-isomer, (L)-isomer, and racemic mixtures thereof and other mixtures, such as enantiomerically or diastereomeric enriched mixtures, all of which belong to the present Within the scope of the invention. Additional asymmetric carbon atoms may be present in the substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are included within the scope of the invention.
  • enantiomer or “optical isomer” refer to stereoisomers that are mirror images of one another.
  • cis-trans isomer or “geometric isomer” is caused by the inability to freely rotate a single bond due to a double bond or a ring-forming carbon atom.
  • diastereomer refers to a stereoisomer in which the molecule has two or more chiral centers and the molecules are in a non-mirrored relationship.
  • wedge-shaped dashed keys Represents the absolute configuration of a solid center with straight solid keys
  • straight dashed keys Indicates the relative configuration of the stereocenter, using wavy lines Indicates a wedge solid key Or wedge-shaped dotted key Or with wavy lines Represents a straight solid key And straight dashed keys
  • a double bond structure exists in a compound, such as a carbon-carbon double bond, a carbon-nitrogen double bond, and a nitrogen-nitrogen double bond, and each atom on the double bond is bonded to two different substituents (including a nitrogen atom) Of the double bonds, a pair of lone pairs of electrons on the nitrogen atom are considered as a substituent to which they are attached), if a wavy line is used between the atom on the double bond and the substituent in the compound
  • the linkage means the (Z) isomer, the (E) isomer or a mixture of the two isomers of the compound.
  • the following formula (A) indicates that the compound exists as a single isomer of the formula (A-1) or the formula (A-2) or two isomers of the formula (A-1) and the formula (A-2) The mixture is present;
  • the following formula (B) indicates that the compound exists as a single isomer of formula (B-1) or formula (B-2) or two of formula (B-1) and formula (B-2) A mixture of isomers is present.
  • the following formula (C) indicates that the compound exists as a single isomer of the formula (C-1) or the formula (C-2) or two isomers of the formula (C-1) and the formula (C-2) The mixture is present.
  • tautomer or “tautomeric form” mean that the different functional isomers are in dynamic equilibrium at room temperature and can be rapidly converted into each other. If tautomers are possible (as in solution), the chemical equilibrium of the tautomers can be achieved.
  • proton tautomers also known as prototropic tautomers
  • prototropic tautomers include interconversions by proton transfer, such as keto-enol isomerization and imine-enes. Amine isomerization.
  • the valence tautomer includes the mutual transformation of some of the bonding electrons.
  • keto-enol tautomerization is the interconversion between two tautomers of pentane-2,4-dione and 4-hydroxypent-3-en-2-one.
  • the terms "enriched in one isomer”, “isomer enriched”, “enriched in one enantiomer” or “enantiomeric enriched” refer to one of the isomers or pairs
  • the content of the oligo is less than 100%, and the content of the isomer or enantiomer is 60% or more, or 70% or more, or 80% or more, or 90% or more, or 95% or more, or 96% or more, or 97% or more, 98% or more, 99% or more, 99.5% or more, 99.6% or more, 99.7% or more, 99.8% or more, or greater than or equal to 99.9%.
  • the term “isomer excess” or “enantiomeric excess” refers to the difference between the two isomers or the relative percentages of the two enantiomers. For example, if one of the isomers or enantiomers is present in an amount of 90% and the other isomer or enantiomer is present in an amount of 10%, the isomer or enantiomeric excess (ee value) is 80%. .
  • optically active (R)- and (S)-isomers as well as the D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If an enantiomer of a compound of the invention is desired, it can be prepared by asymmetric synthesis or by derivatization with a chiral auxiliary wherein the resulting mixture of diastereomers is separated and the auxiliary group cleaved to provide pure The desired enantiomer.
  • a diastereomeric salt is formed with a suitable optically active acid or base, followed by conventional methods well known in the art.
  • the diastereomers are resolved and the pure enantiomer is recovered.
  • the separation of enantiomers and diastereomers is generally accomplished by the use of chromatography using a chiral stationary phase, optionally in combination with chemical derivatization (eg, formation of an amino group from an amine). Formate).
  • the compounds of the invention may contain unnatural proportions of atomic isotopes on one or more of the atoms that make up the compound.
  • radiolabeled compounds can be used, such as tritium (3 H), iodine -125 (125 I) or C-14 (14 C).
  • hydrogen can be replaced by heavy hydrogen to form a deuterated drug.
  • the bond composed of barium and carbon is stronger than the bond composed of common hydrogen and carbon.
  • deuterated drugs have reduced side effects and increased drug stability. Enhance the efficacy and prolong the biological half-life of the drug. Alterations of all isotopic compositions of the compounds of the invention, whether radioactive or not, are included within the scope of the invention.
  • substituted means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, and may include variants of heavy hydrogen and hydrogen, as long as the valence of the particular atom is normal and the substituted compound is stable. of.
  • Oxygen substitution does not occur on the aromatic group.
  • optionally substituted means that it may or may not be substituted, and unless otherwise specified, the kind and number of substituents may be arbitrary on the basis of chemically achievable.
  • any variable eg, R
  • its definition in each case is independent.
  • the group may optionally be substituted with at most two R, and each case has an independent option.
  • combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds.
  • linking group When the number of one linking group is 0, such as -(CRR) 0 -, it indicates that the linking group is a single bond.
  • one of the variables When one of the variables is selected from a single bond, it means that the two groups to which it is attached are directly linked. For example, when L represents a single bond in A-L-Z, the structure is actually A-Z.
  • substituent When a substituent is vacant, it means that the substituent is absent.
  • X when X is vacant in A-X, the structure is actually A.
  • substituents are not indicated by which atom is attached to the substituted group, such a substituent may be bonded through any atom thereof, for example, a pyridyl group as a substituent may be passed through any one of the pyridine rings. A carbon atom is attached to the substituted group.
  • the medium linking group L is -MW-, and at this time, -MW- can be connected in the same direction as the reading order from left to right to form ring A and ring B. It is also possible to connect the ring A and the ring B in a direction opposite to the reading order from left to right. Combinations of the linking groups, substituents and/or variants thereof are permissible only if such combinations result in stable compounds.
  • any one or more of the sites may be attached to the other group via a chemical bond.
  • the chemical bond connecting the site to other groups can be a straight solid bond Straight dotted key Or wavy line Said.
  • a straight solid bond in -OCH 3 indicates attachment to other groups through an oxygen atom in the group;
  • a straight dashed bond in the middle indicates that both ends of the nitrogen atom in the group are attached to other groups;
  • the wavy line in the middle indicates that it is attached to other groups through the carbon atoms at the 1 and 2 positions in the phenyl group.
  • hetero denotes a hetero atom or a hetero atomic group (ie, a radical containing a hetero atom), including atoms other than carbon (C) and hydrogen (H), and radicals containing such heteroatoms, including, for example, oxygen (O). ), nitrogen (N), sulfur (S) or -N (H)-.
  • the number of atoms on the ring is generally defined as the number of elements of the ring.
  • “5-7 membered ring” refers to a “ring” that is arranged around 5-7 atoms.
  • ring denotes cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, cycloalkynyl, heterocycloalkynyl, aryl or heteroaryl.
  • the ring includes a single ring, and also includes a bicyclic or polycyclic ring system such as a spiro ring, a ring and a bridge ring. Unless otherwise specified, the ring optionally contains from 1 to 3 heteroatoms.
  • a "5-7 membered ring” includes, for example, phenyl, pyridyl and piperidinyl; on the other hand, the term “5-7 membered heterocycloalkyl” includes piperidinyl, but does not include phenyl.
  • the term “ring” also includes ring systems containing at least one ring, each of which "ring” independently conforms to the above definition.
  • C1-6 alkyl is used to indicate a straight or branched saturated hydrocarbon group containing from 1 to 6 carbon atoms.
  • the C 1-6 alkyl group includes C 1-5 , C 1-4 , C 1-3 and C 1-2 alkyl groups and the like. It may be monovalent (such as methyl), divalent (such as methylene) or multivalent (such as methine).
  • C 1-6 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl) , s-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl and the like.
  • heteroalkyl denotes a stable straight or branched alkyl radical or a combination thereof consisting of a number of carbon atoms and at least one heteroatom or heteroatom. Things.
  • the heteroatoms are selected from the group consisting of O, N, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen heteroatoms are optionally quaternized.
  • the heteroalkyl group is a C1-6 heteroalkyl group; in other embodiments, the heteroalkyl group is a C1-3 heteroalkyl group.
  • the heteroatom or heteroatom group may be located at any internal position of the heteroalkyl group, including the position at which the alkyl group is attached to the rest of the molecule, but the term " C1-6 alkylamino" refers to those in which the amino group is attached to the remainder of the molecule.
  • the C 1-6 alkylamino group includes C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 1 , C 2 , C 3 , C 4 , C 5 , C 6 alkylamino groups and the like.
  • Examples of the C 1-6 alkylamino group include, but are not limited to, -NHCH 3 , -N(CH 3 ) 2 , -NHCH 2 CH 3 , -N(CH 3 )(CH 2 CH 3 ), and the like.
  • Up to two heteroatoms
  • C 1-3 alkylamino refers to those alkyl groups containing from 1 to 3 carbon atoms which are attached to the remainder of the molecule.
  • the C 1-3 alkylamino group includes C 1-2 , C 1 , C 2 , C 3 alkylamino group and the like.
  • Examples of the C 1-3 alkylamino group include, but are not limited to, -NHCH 3 , -N(CH 3 ) 2 , -NHCH 2 CH 3 , -N(CH 3 )(CH 2 CH 3 ), and the like.
  • C 3-6 cycloalkyl is used to mean any stable cyclic alkyl group containing 3 to 6 carbon atoms, the C 3-6 cycloalkyl is a monocyclic ring system.
  • the C 3-6 cycloalkyl group includes C 3-5 , C 3-4 , C 4-6 , C 5-6 , C 3 and C 6 cycloalkyl groups and the like, and it may be monovalent, divalent or Multi-price.
  • Examples of C 3-6 cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.
  • the 4-8 membered heterocycloalkyl group includes 4-5 members, 4-6 members, 5-6 members, 5-membered and 6-membered heterocycloalkyl groups, and the like.
  • Examples of 4-8 membered heterocycloalkyl include, but are not limited to, azetidinyl, oxetanyl, thioheterobutyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl ( Including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc., tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2- Piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl
  • a hetero atom may occupy a position where a heterocycloalkyl group is bonded to the rest of the molecule.
  • the 5-6 membered heterocycloalkyl group includes 5-membered and 6-membered heterocycloalkyl groups.
  • 5-6 membered heterocycloalkyl examples include, but are not limited to, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.) , tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1 - piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxoalkyl, dithiaalkyl, isoxazolidinyl, isothiazole Alkyl, 1,2-oxazinyl, 1,2-thiaziny
  • C n-n+m or C n -C n+m includes any one of n to n+m carbons, for example, C 1-12 includes C 1 , C 2 , C 3 , C 4 , C 5 , C 6 , C 7 , C 8 , C 9 , C 10 , C 11 , and C 12 , also including any range of n to n+m, for example, C 1-12 includes C 1 - 3 , C 1-6 , C 1-9 , C 3-6 , C 3-9 , C 3-12 , C 6-9 , C 6-12 , and C 9-12 , etc.; similarly, n to n
  • the +m element indicates that the number of atoms on the ring is n to n+m, for example, the 3-12 element ring includes a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-
  • a 10-membered ring, a 11-membered ring, and a 12-membered ring and includes any one of n to n+m, for example, a 3-12-membered ring including a 3-6-membered ring, a 3-9-membered ring, and a 5-6-membered ring. Ring, 5-7 membered ring, 6-7 membered ring, 6-8 membered ring, and 6-10 membered ring.
  • leaving group refers to a functional group or atom which may be substituted by another functional group or atom by a substitution reaction (for example, an affinity substitution reaction).
  • substituent groups include triflate; chlorine, bromine, iodine; sulfonate groups such as mesylate, tosylate, p-bromobenzenesulfonate, p-toluenesulfonic acid Esters and the like; acyloxy groups such as acetoxy, trifluoroacetoxy and the like.
  • protecting group includes, but is not limited to, "amino protecting group", “hydroxy protecting group” or “thiol protecting group”.
  • amino protecting group refers to a protecting group suitable for preventing side reactions at the amino nitrogen position.
  • Representative amino protecting groups include, but are not limited to, formyl; acyl, such as alkanoyl (e.g., acetyl, trichloroacetyl or trifluoroacetyl); alkoxycarbonyl, e.g., tert-butoxycarbonyl (Boc) Arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), trityl (Tr), 1, 1-di -(4'-methoxyphenyl)methyl; silyl groups such as trimethylsilyl (TMS) and tert-
  • hydroxy protecting group refers to a protecting group suitable for use in preventing hydroxy side reactions.
  • Representative hydroxy protecting groups include, but are not limited to, alkyl groups such as methyl, ethyl and t-butyl groups; acyl groups such as alkanoyl groups (e.g., acetyl); arylmethyl groups such as benzyl (Bn), Oxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl groups such as trimethylsilyl (TMS) and tert-butyl Dimethylsilyl (TBS) and the like.
  • alkyl groups such as methyl, ethyl and t-butyl groups
  • acyl groups such as alkanoyl groups (e.g., acetyl)
  • arylmethyl groups such as benzyl (Bn), Oxybenzyl (PMB), 9-fluoreny
  • the compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments set forth below, combinations thereof with other chemical synthetic methods, and those well known to those skilled in the art. Equivalent alternatives, preferred embodiments include, but are not limited to, embodiments of the invention.
  • the solvent used in the present invention is commercially available.
  • the present invention employs the following abbreviations: MeOH for methanol; DMSO for dimethyl sulfoxide; and CDCl 3 for deuterated chloroform.
  • the compounds of the invention are based on conventional nomenclature or use in the art Software naming, commercially available compounds using the supplier catalog name.
  • Figure 1 shows the inhibition of ⁇ -SMA gene expression by compounds 11, 12 (hydrochloride) and 13 (hydrochloride) of the present invention
  • Figure 2 is a pulmonary fibrosis score of compounds 11 and 12 (hydrochloride) of the present invention
  • Figure 3 shows the damage of bronchial and arterioles in the lesions of compounds 11 and 12 (hydrochloride) of the present invention
  • Figure 4 shows the damage of bronchial and arterioles at the margin of the lesion by compounds 11 and 12 (hydrochloride) of the present invention.
  • hydrochloride or formate salt of the compound of the present invention is adjusted to pH neutrality by adding a saturated sodium hydrogencarbonate solution, and is separated by high performance liquid chromatography (neutral, ammonium hydrogencarbonate system) to obtain a free base of the compound.
  • Trifluoroacetic acid (193 mg, 0.13 mL) was added to a solution of compound 5a (50.0 mg, 0.170 mmol) in dichloromethane (2 mL). After the addition was completed, the reaction solution was heated to 25 ° C and stirred for 12 hours. After completion of the reaction, a saturated aqueous solution of sodium hydrogen carbonate (20 mL) was evaporated and evaporated. The combined organic phases were washed with brine (20 mL EtOAc) After the desiccant was filtered off, the solvent was removed under reduced pressure and the residue was purified by preparative chromatography chromatography to afford compound 5b.
  • the compound 21b was obtained in the first step of Example 6.
  • the compound 21d was obtained by referring to the third step of Example 6.
  • the compound 21e was obtained by referring to the fourth step of Example 6.
  • buffer solution 20 mM 4-hydroxyethylpiperazine ethanesulfuric acid (pH 7.5), 10 mM magnesium chloride, 1 mM ethylene glycol diethyl ether diamine tetraacetic acid, 0.02% polyoxyethylene lauryl ether, 0.02 mg / mL bovine serum albumin , 0.1 mM sodium vanadate, 2 mM dithiothreitol, 1% DMSO.
  • the freshly prepared buffer solution was added to a ROCK protein kinase substrate Long S6 Kinase substrate, at a concentration of 20 ⁇ M, and then 1 nM ROCK protein kinase was added thereto, and uniformly stirred.
  • a series of DMSO dilutions containing the test compound (starting at 10 ⁇ M, serially diluted 3 times) were added using Echo 550, pre-incubated for 20 minutes at room temperature, and 33 P-ATP (radiation intensity 10 ⁇ Ci/ ⁇ L) was added to initiate the reaction. hour. It was then filtered using P81 ion exchange paper (Whatman #3698-915) and washed with 0.75% phosphoric acid. The intensity of the radiation was measured using the Filter-Binding method.
  • the protein kinase inhibitory activity of the compound is expressed as protein kinase activity remaining relative to the blank substrate (DMSO alone). And the values of IC 50 was calculated using the Prism software package curve (GraphPad Software, San Diego California, USA).
  • the compound of the present invention has high inhibitory activity against ROCK2 and has certain selective ROCK2 inhibitory activity.
  • NIH-3T3 cells were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences.
  • the TRIzol reagent was purchased from Infineon, and the genomic DNA reverse transcription kit was purchased from Tiangen Biochemical.
  • the real-time fluorescent quantitative PCR premix was purchased from Nanjing Nuoweizan, isopropanol and chloroform from the Chinese medicine.
  • NIH-3T3 cells with a growth confluence of 80% were digested with a 0.25% trypsin into a single cell suspension and plated in 6-well plates.
  • the cell plates were placed at 37 °, overnight culture incubator in 5% CO 2.
  • the compound was prepared the next day, and 10 mM of the compound (11, 12, 13) to be tested was diluted to 200 ⁇ M, and added to a 6-well plate at 10 ⁇ L per well, and cultured overnight.
  • the final concentration of the compound was 1 ⁇ M and the final concentration of DMSO was 1%.
  • the reaction conditions were 50 ° C for 2 minutes, 95 ° C for 10 minutes, 95 ° C for 30 seconds, 60 ° C for 30 seconds, 95 ° C for 30 seconds, and 60 ° C for 30 seconds for 40 cycles.
  • the CT value of the reaction was derivatized at the end of the reaction to calculate the relative expression amount of the sample. data analysis:
  • the relative gene expression amount was calculated by the 2 - ⁇ CT calculation method, and a significant analysis was performed using a two-tailed T test (the compound test concentration was 1 ⁇ M).
  • the compounds of the present invention have a good inhibitory activity on the expression of ⁇ -SMA gene.
  • Plasma samples were administered orally by two other rats at a dose of 2 mg/kg, and plasma samples of 0.25, 0.5, 1, 2, 3, 4, 6, 8, 24 hours after administration were collected. Collect whole blood samples within 24 hours, centrifuge at 3000g for 15 minutes, separate the supernatant to obtain plasma samples, add the internal standard acetonitrile solution to precipitate the protein, mix thoroughly and centrifuge to take the supernatant, and quantify by LC-MS/MS analysis. Analyze blood drug concentration and calculate pharmacokinetic parameters such as peak concentration (C max ), clearance rate (CL), half-life (T 1/2 ), tissue distribution (Vdss), area under the drug-time curve (AUC 0-last) ), bioavailability (F), etc.
  • C max peak concentration
  • CL clearance rate
  • T 1/2 half-life
  • Vdss tissue distribution
  • AUC 0-last area under the drug-time curve
  • bioavailability F
  • the compounds of the invention have good pharmacokinetic properties, including good oral bioavailability, oral exposure, half-life and clearance.
  • Model Left unilateral pulmonary pulmonary fibrosis model in SD rats: Rats were injected with bleomycin (3 mg/kg in 1.5 mL/kg) to induce pulmonary fibrosis.
  • bleomycin bleomycin
  • modeling was started 4 days after adaptive feeding. Animals were weighed and inhaled with isoflurane. After confirming the anesthesia of the animals, the neck was disinfected, the skin of the neck was cut, the main air tube was exposed to the muscles, and a small opening was made between the tracheal rings. The left main bronchus was directly injected into bleomycin, and the trachea and skin were sutured. After the operation, the animals were placed in a 37 ° C electric blanket to keep the animals fully awake, and it was confirmed that the animals were able to return to the cages for normal feeding after free feeding and drinking. Oral administration of the compound was started on the 8th day of modeling and continued for 14 days. 24 hours after the end of the last dose, all animals were euthanized; the lungs were collected by systemic perfusion with low temperature PBS, and replenished with formalin, and the tissue was fixed in 10% formalin solution for subsequent histopathology. analysis.
  • Pathological examination of the left lung H&E staining pathological evaluation: 1) pathological changes of the left lung terminal bronchioles and arterioles, 2) left lung pulmonary fibrosis area, 3) left lung pulmonary fibrosis Ashcroft score.
  • the compounds of the present invention have a significant improvement in bronchial and arteriolar lesions on the inside of the lesion or on the edge of the lesion ( Figures 3 and 4), and also reduce the pulmonary fibrosis score (Figure 2), at a lower level. At doses (10mpk), it is comparable to nidanib (100mpk).

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pulmonology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention concerne un composé de pyrazole tenant lieu d'inhibiteur de la RHO-kinase, sa composition pharmaceutique et ses applications dans la préparation de cette dernière. L'invention concerne un médicament inhibant RHO et plus particulièrement ledit composé de formule (I.1), son sel pharmaceutiquement acceptable et son isomère.
PCT/CN2019/083208 2018-04-18 2019-04-18 Composé de pyrazole tenant lieu d'inhibiteur de la rho-kinase WO2019201296A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201810349378 2018-04-18
CN201810349378.0 2018-04-18

Publications (1)

Publication Number Publication Date
WO2019201296A1 true WO2019201296A1 (fr) 2019-10-24

Family

ID=68239916

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2019/083208 WO2019201296A1 (fr) 2018-04-18 2019-04-18 Composé de pyrazole tenant lieu d'inhibiteur de la rho-kinase

Country Status (1)

Country Link
WO (1) WO2019201296A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021093795A1 (fr) * 2019-11-15 2021-05-20 武汉朗来科技发展有限公司 Inhibiteur de rock, son procédé de préparation et son utilisation
US11078161B2 (en) * 2017-06-16 2021-08-03 Hitgen Inc. Rock-inhibiting compound and uses thereof

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105085525A (zh) * 2014-04-28 2015-11-25 南京明德新药研发股份有限公司 作为rho激酶抑制剂的异喹啉磺酰衍生物
WO2017091661A1 (fr) * 2015-11-25 2017-06-01 Strovel Jeffrey William Inhibiteurs de bromodomaines bet bicycliques et leurs utilisations
WO2018081108A1 (fr) * 2016-10-24 2018-05-03 Translational Drug Development, Llc Composés amides en tant qu'inhibiteurs de kinase

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105085525A (zh) * 2014-04-28 2015-11-25 南京明德新药研发股份有限公司 作为rho激酶抑制剂的异喹啉磺酰衍生物
WO2017091661A1 (fr) * 2015-11-25 2017-06-01 Strovel Jeffrey William Inhibiteurs de bromodomaines bet bicycliques et leurs utilisations
WO2018081108A1 (fr) * 2016-10-24 2018-05-03 Translational Drug Development, Llc Composés amides en tant qu'inhibiteurs de kinase

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11078161B2 (en) * 2017-06-16 2021-08-03 Hitgen Inc. Rock-inhibiting compound and uses thereof
WO2021093795A1 (fr) * 2019-11-15 2021-05-20 武汉朗来科技发展有限公司 Inhibiteur de rock, son procédé de préparation et son utilisation

Similar Documents

Publication Publication Date Title
ES2426482T3 (es) Inhibidor de IGF-1R
CN110300589B (zh) 作为cftr增效剂的双环异杂芳基衍生物
JP2022071072A (ja) （ｓ）－７－（１－アクリロイルピペリジン－４－イル）－２－（４－フェノキシフェニル）－４，５，６，７－テトラ－ヒドロピラゾロ［１，５－ａ］ピリミジン－３－カルボキサミドの結晶形、その調製、及びその使用
JP6948659B1 (ja) ピリダジニルチアアゾールカルボキシアミド化合物
CN109970745B (zh) 取代的吡咯并三嗪类化合物及其药物组合物及其用途
TW201625626A (zh) 作爲蛋白質激酶抑制劑之胺基噠嗪酮化合物
CN106232599A (zh) 4‑氨基‑咪唑并喹啉化合物
BR112015028501B1 (pt) Compostos derivados de bipirazol, seus sais, composição compreendendo o composto ou o sal, método de inibição in vitro de uma atividade de jak1, e processo de preparação de sal de ácido fosfórico
US20100204187A1 (en) Purine Derivatives
BR112018001469B1 (pt) Composto, composição farmacêutica e uso de um composto
WO2019201297A1 (fr) Composé de benzo-pyrazole tenant lieu d'inhibiteur de la rho-kinase
WO2017092413A1 (fr) Composés diaminopyrimidine et composition comprenant ceux-ci
TW201315727A (zh) 尿嘧啶衍生物及其醫藥用途
EA025840B1 (ru) Производные пиразина в качестве агонистов каннабиноидного рецептора 2
JP7409719B2 (ja) Ｅｒｋ阻害剤としてのチアゾロラクタム系化合物およびその使用
JP7397452B2 (ja) ヘテロアリールカルボキシアミド化合物
WO2019201296A1 (fr) Composé de pyrazole tenant lieu d'inhibiteur de la rho-kinase
CN115413279A (zh) P2x3调节剂
WO2023041055A1 (fr) Inhibiteur de kif18a
WO2000043385A1 (fr) Composes heterocycliques et agents antitumoraux les comprenant en tant que principe actif
WO2021129841A1 (fr) Composé utilisé comme inhibiteur de kinase ret et son application
JP3588363B2 (ja) キノキサリンジオン類
EP3936507A1 (fr) Composés ayant à la fois des effets d'inhibition de protéine de bromodomaine bet et de régulation de gène pd-l1
WO2019174576A1 (fr) Composé imidaxopyrolone et son application
AP982A (en) Quinoxalinediones.

Legal Events

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

Ref document number: 19788523

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19788523

Country of ref document: EP

Kind code of ref document: A1

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205A DATED 21/04/2021)

122 Ep: pct application non-entry in european phase

Ref document number: 19788523

Country of ref document: EP

Kind code of ref document: A1