WO2024111626A1 - Nouveau dérivé de thiazole - Google Patents

Nouveau dérivé de thiazole Download PDF

Info

Publication number
WO2024111626A1
WO2024111626A1 PCT/JP2023/041996 JP2023041996W WO2024111626A1 WO 2024111626 A1 WO2024111626 A1 WO 2024111626A1 JP 2023041996 W JP2023041996 W JP 2023041996W WO 2024111626 A1 WO2024111626 A1 WO 2024111626A1
Authority
WO
WIPO (PCT)
Prior art keywords
group
compound
reaction
amino
pyrazol
Prior art date
Application number
PCT/JP2023/041996
Other languages
English (en)
Japanese (ja)
Inventor
匡明 澤
真以 新井
充治 花田
大本　弘志
Original Assignee
カルナバイオサイエンス株式会社
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 カルナバイオサイエンス株式会社 filed Critical カルナバイオサイエンス株式会社
Publication of WO2024111626A1 publication Critical patent/WO2024111626A1/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
    • A61K31/425—Thiazoles
    • A61K31/427—Thiazoles not condensed 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
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
    • 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
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/454—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone
    • 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
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/4545—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring hetero atom, e.g. pipamperone, anabasine
    • 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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • 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
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings

Definitions

  • the present invention relates to a medicine, in particular to a novel thiazole derivative or a pharma- ceutical acceptable salt thereof that has TGF ⁇ type I receptor (ALK5) inhibitory activity.
  • TGF- ⁇ Transforming growth factor ⁇
  • TGF- ⁇ a multifunctional cytokine
  • TGF- ⁇ 1, ⁇ 2, ⁇ 3 three isoforms in mammals and is involved in physiological functions such as cell proliferation, differentiation control, migration, and adhesion
  • TGF- ⁇ binds to a complex formed by a single-pass transmembrane type I receptor (ALK5) with a serine/threonine kinase domain and a type II receptor, and transmits a signal into the cell.
  • ALK5 is activated by the type II receptor upon binding of TGF- ⁇ , ALK5 phosphorylates the transcription factors Smad2 and Smad3 in the cell.
  • Non-Patent Documents 1 and 2 After forming a complex with Smad4, phosphorylated Smad2 and Smad3 migrate to the nucleus and induce the transcription of target genes directly or together with other transcription factors (Non-Patent Documents 1 and 2).
  • TGF- ⁇ signaling plays an important role in pathological conditions such as wound healing, inflammation/immunity, and cancer invasion/metastasis (Non-Patent Document 3).
  • EMT epithelial-mesenchymal transition
  • angiogenesis angiogenesis
  • TGF- ⁇ is involved in the proliferation of osteosarcoma
  • ALK5 inhibitors suppress the proliferation of osteosarcoma cells stimulated with TGF- ⁇
  • anti-TGF- ⁇ antibodies suppress lung metastasis of osteosarcoma
  • ALK5 inhibitors are useful for the treatment of solid cancers and metastasis in which TGF- ⁇ is involved.
  • TGF- ⁇ controls the differentiation induction of regulatory T cells (Treg), which are responsible for immune control, and it is thought that in the cancer microenvironment, Treg cells suppress antitumor immunity and promote tumor growth (Non-Patent Document 8). Therefore, suppressing TGF- ⁇ signaling is expected to have an antitumor effect by cancer immunotherapy and a combined effect with immune checkpoint inhibitors.
  • TGF- ⁇ is also involved in fibrosis, a pathological excess state of the normal tissue repair process, and it has been reported that abnormal excess signaling of TGF- ⁇ is the cause of fibrosis (Non-Patent Document 9). Therefore, a compound having an ALK5 inhibitory activity is useful for treating diseases in which TGF- ⁇ signaling is involved, such as cancer, and for expanding and enhancing the therapeutic effect in cancer immunotherapy by using it in combination with an immune checkpoint inhibitor, etc. It is also useful for treating and preventing fibrotic diseases, etc.
  • the objective of the present invention is to provide a pharmaceutical, in particular a novel thiazole derivative or a pharma- ceutical acceptable salt thereof that has TGF ⁇ type I receptor (ALK5) inhibitory activity.
  • the present invention can be achieved by the following thiazole derivatives or pharma- ceutically acceptable salts thereof: (1) The following formula (I): (wherein R 1 represents an optionally substituted alkyl group, an optionally substituted saturated heterocyclic group or an optionally substituted amino group; R 2 represents an optionally substituted aryl group, an optionally substituted heteroaryl group or an optionally substituted alkynyl group; and Z represents an optionally substituted alkyl group, an optionally substituted alkenyl group, an optionally substituted alkynyl group, an optionally substituted saturated heterocyclic group or an optionally substituted cycloalkyl group), or a pharmacy acceptable salt thereof.
  • R 1 represents an optionally substituted alkyl group, an optionally substituted saturated heterocyclic group or an optionally substituted amino group
  • R 2 represents an optionally substituted aryl group, an optionally substituted heteroaryl group or an optionally substituted alkynyl group
  • Z represents an optionally substituted alky
  • the present inventors conducted various studies to solve the above problems, and found that the novel thiazole derivative represented by the above formula (I) and its pharma- ceutical acceptable salt exhibit ALK5 inhibitory activity, and thus completed the present invention.
  • the compounds provided by the present invention are useful for treating diseases known to be associated with abnormal cell responses via the TGF ⁇ signal pathway, particularly cancer, and are also useful for preventing tumor metastasis and recurrence by targeting cancer stem cells. Furthermore, by using them in combination with immune checkpoint inhibitors, they are useful for expanding and enhancing the therapeutic effect in cancer immunotherapy. They are also useful for treating and preventing fibrotic diseases and the like. They are also useful as experimental and research reagents as ALK5 inhibitors and TGF ⁇ signal inhibitors.
  • the novel thiazole derivative of the present invention has the following formula (I):
  • R1 represents an alkyl group which may have a substituent, a saturated heterocyclic group which may have a substituent, or an amino group which may have a substituent
  • R2 represents an aryl group which may have a substituent, a heteroaryl group which may have a substituent, or an alkynyl group which may have a substituent
  • Z represents an alkyl group which may have a substituent, an alkenyl group which may have a substituent, an alkynyl group which may have a substituent, a saturated heterocyclic group which may have a substituent, or a cycloalkyl group which may have a substituent.
  • It is a compound represented by the formula:
  • halogen means a fluorine atom, a chlorine atom, a bromine atom, or an iodine atom.
  • C 1-6 alkyl means a linear or branched saturated hydrocarbon group having 1-6 carbon atoms
  • C 6 alkyl means a saturated hydrocarbon group having 6 carbon atoms. The same applies to other numbers.
  • C 1-3 alkyl specifically means a methyl group, an ethyl group, a propyl group, or an isopropyl group.
  • examples of “C 1-6 alkyl” include a butyl group, a 1-methylpropyl group, a 2-methylpropyl group, a tert-butyl group, a pentyl group, a 1,1-dimethylpropyl group, a 1,2-dimethylpropyl group, a 1-methylbutyl group, a 2-methylbutyl group, a 4-methylpentyl group, a 3-methylpentyl group, a 2-methylpentyl group, a 1-methylpentyl group, and an n-hexyl group.
  • C 3-8 cycloalkyl refers to a cyclic saturated hydrocarbon group having 3 to 8 carbon atoms.
  • Specific examples include a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, and a cyclooctyl group.
  • C 1-6 alkoxy means “C 1-6 alkyloxy”
  • the “C 1-6 alkyl” portion has the same meaning as the above-mentioned "C 1-6 alkyl”.
  • the substituents may be one or more of any type of substituent at any chemically possible position, and when there are two or more substituents, the respective substituents may be the same or different.
  • R1 R 1 represents an alkyl group which may have a substituent, a saturated heterocyclic group which may have a substituent, or an amino group which may have a substituent.
  • the alkyl group moiety of the alkyl group which may have a substituent may be any of C1-6 alkyl, and particularly preferably C1-3 alkyl.
  • a specific example of a preferred substituent in the C 1-6 alkyl or C 1-3 alkyl is a substituted amino group --NR 11 R 12.
  • R 11 and R 12 each independently represent , a 4- to 6-membered oxygen-containing saturated heterocycle, or a C 1-6 alkyl, or a 4- to 6-membered ring which, taken together with the nitrogen atom to which it is bonded, may further contain a nitrogen atom or an oxygen atom.
  • the oxygen-containing saturated heterocycle include an oxetanyl group, a tetrahydropyranyl group, and a tetrahydrofuryl group
  • examples of the nitrogen-containing saturated heterocycle include pyrrolidine, piperidine, piperazine, and morpholine. These may be substituted with halogen, a methyl group, a methoxy group, or the like.
  • a preferred embodiment of the alkyl group which may have a substituent is -CH 2 NR 11 R 12 in which the above-mentioned substituted amino group is substituted with a methyl group.
  • the saturated heterocyclic group portion of the optionally substituted saturated heterocyclic group is a 4- to 6-membered heterocyclic group containing 1 to 3 heteroatoms arbitrarily selected from a sulfur atom, a nitrogen atom, and an oxygen atom. and can be substituted at any position where chemical substitution is possible.
  • the substituent can be an azetidinyl group, a pyrrolidinyl group, a piperidinyl group, a morpholinyl group, a piperazinyl group, an oxetanyl group, or , tetrahydrofuranyl group, tetrahydrothienyl group, tetrahydropyranyl group, thiomorpholinyl group, etc.
  • the substituent of the saturated heterocyclic group may be a halogen atom, a hydroxy group, a methyl group, a methoxy group, a methanesulfonyl group, etc.
  • Examples include a 2-hydroxyethyl group, an oxetanyl group, a tetrahydropyranyl group, a methyl-piperidinyl group, a methyl-piperazinyl group, a morpholinyl group, and a thiomorpholinyl group.
  • the amino group which may have a substituent in R 1 is represented by the atomic group -NR 13 R 14 , R 13 and R 14 each independently represent a C 1-6 alkyl, The 6- alkyl may be further substituted with a dimethylamino group, a methoxy group, etc.
  • a preferred embodiment of the amino group which may have a substituent is a dimethylamino group.
  • R2 R2 represents an aryl group which may have a substituent, a heteroaryl group which may have a substituent, or an alkynyl group which may have a substituent.
  • Examples of the aryl group moiety of the aryl group which may have a substituent include a phenyl group and a naphthyl group.
  • the heteroaryl group portion of the heteroaryl group which may have a substituent is a 4- to 6-membered heteroaryl ring containing 1 to 4 heteroatoms arbitrarily selected from a sulfur atom, a nitrogen atom, and an oxygen atom.
  • the formula includes a pyrrolyl group, a furanyl group, a thienyl group, an imidazolyl group, a pyrazolyl group, an oxazolyl group, a thiazolyl group, an imidazolyl group, a triazolyl group, a tetrazolyl group, a pyridyl group, a pyrazinyl group, a pyrimidinyl group, a pyridazinyl group, a triazinyl group, etc. are examples.
  • the alkynyl group portion of the alkynyl group which may have a substituent is a straight-chain or branched-chain unsaturated hydrocarbon having at least one carbon-carbon triple bond and 2 to 6 carbon atoms. Specifically, it means an ethynyl group, a 2-propynyl group, a 2-butynyl group, a 2-pentynyl group, a 3-pentynyl group, a 2-hexynyl group, a 3-hexynyl group. etc. are examples.
  • the substituents in the above-mentioned aryl group which may have a substituent, the heteroaryl group which may have a substituent, and the alkynyl group which may have a substituent are preferably C1
  • the substituents of the C 1-6 alkyl and C 1-6 alkoxy include halogen, cyano, hydroxyl, and benzyl.
  • the substituents include 1 to 3 halogens, a hydroxyl group, a cyclopropyl group, a phenyl group, etc.
  • Examples of the substituents that form a ring include a methylenedioxy group or an ethylenedioxy group. Examples can be given.
  • Z Z represents an alkyl group which may have a substituent, an alkenyl group which may have a substituent, an alkynyl group which may have a substituent, a saturated heterocyclic group which may have a substituent, or a cycloalkyl group which may have a substituent.
  • the alkyl group portion of the alkyl group which may have a substituent is a C 1-6 alkyl.
  • Examples of the substituent of this C 1-6 alkyl include a C 3-6 cycloalkyl group, a halogeno C 3-6 cycloalkyl group, a C 1-3 alkoxy group, a halogen atom, a hydroxy group, a benzyloxy group, a methylamino group, a dimethylamino group, a pyranyl group, a piperidinyl group, and a C 1-3 alkylsulfonyl group.
  • the alkenyl group moiety of the alkenyl group which may have a substituent may be any of linear or branched alkenyl groups having 2 to 6 carbon atoms, such as an allyl group, a 2-butenyl group, an isobutenyl group, a 2-pentenyl group, a 3-pentenyl group, and a 2-hexenyl group.
  • the alkynyl group which may have a substituent in Z is the same as the alkynyl group which may have a substituent in R2 .
  • the cycloalkyl group portion of the cycloalkyl group which may have a substituent may be any cyclic saturated hydrocarbon group having 3 to 8 carbon atoms (C cycloalkyl ), with a cyclopropyl group being particularly preferred.
  • Examples of the substituent of this C cycloalkyl include halogen atoms.
  • the saturated heterocyclic group portion of the optionally substituted saturated heterocyclic group may be any 4- to 6-membered saturated heterocyclic group containing 1 to 3 heteroatoms arbitrarily selected from sulfur atoms, nitrogen atoms, and oxygen atoms, and may be substituted at any chemically substitutable position.
  • pyrrolidinyl group examples include a pyrrolidinyl group, a piperidinyl group, a morpholinyl group, a piperazinyl group, an oxetanyl group, a tetrahydrofuranyl group, a tetrahydrothienyl group, a tetrahydropyranyl group, a thiomorpholinyl group, and the like.
  • An embodiment of the compound (I) of the present invention is the above thiazole derivative (I) or a pharma- ceutically acceptable salt thereof, wherein Z is a cycloalkyl group which may be substituted.
  • An embodiment of compound (I) of the present invention is the above thiazole derivative (I) or a pharma- ceutically acceptable salt thereof, wherein Z is an alkyl group which may have a substituent.
  • Another embodiment of the compound (I) of the present invention is the above thiazole derivative (I) or a pharma- ceutically acceptable salt thereof, wherein R2 is a phenyl group which may be substituted.
  • Another embodiment of the compound (I) of the present invention is the above thiazole derivative (I) or a pharma- ceutically acceptable salt thereof, wherein R2 is an alkynyl group which may be substituted.
  • Another embodiment of the compound (I) of the present invention is the thiazole derivative (I) or a pharma- ceutically acceptable salt thereof, which is selected from the group consisting of Examples 1 to 226.
  • Compound (I) of the present invention may have isomers depending on, for example, the type of substituent.
  • the chemical structure of only one form of the isomer may be described, but the present invention also includes all isomers (geometric isomers, optical isomers, tautomers, etc.) that may occur due to the structure, and also includes a single isomer or a mixture thereof.
  • Pharmaceutically acceptable salts of compound (I) of the present invention include inorganic acid salts with hydrochloric acid, sulfuric acid, carbonic acid, phosphoric acid, etc., and organic acid salts with formic acid, fumaric acid, maleic acid, methanesulfonic acid, p-toluenesulfonic acid, etc.
  • the present invention also includes alkali metal salts with sodium, potassium, etc., alkaline earth metal salts with magnesium, calcium, etc., organic amine salts with lower alkylamines, lower alcoholamines, etc., basic amino acid salts with lysine, arginine, ornithine, etc., as well as ammonium salts.
  • Compound (I) of the present invention and its pharma- ceutical acceptable salts can be prepared, for example, by the following method. In the preparation methods shown below, if the defined groups change under the conditions of the method or are not suitable for carrying out the method, the compound can be easily prepared by applying a method commonly used in organic synthesis chemistry, such as protection and deprotection of functional groups [T. W. Greene, Protective Groups in Organic Synthesis 4th Edition, John Wiley & Sons, Inc., 2007]. In addition, the order of reaction steps such as introduction of substituents can be changed as necessary.
  • the compound (I) of the present invention can be produced by Ullmann condensation reaction or nucleophilic substitution reaction between the compound (II) and a compound R 2 -X. That is, when R 2 is an aryl group which may have a substituent or a heteroaryl group which may have a substituent, a mixture of compound (II) and compound R 2 -X is subjected to Ullmann condensation reaction in a solvent in the presence of a metal catalyst such as copper, using a base and an additive as necessary, to produce compound (I).
  • the solvent used in the reaction may be any solvent inert to the reaction, and is not particularly limited, but preferably includes dioxane, toluene, dimethoxyethane, DMF, and the like.
  • Compound R 2 -X is preferably used in a molar equivalent or excess amount relative to compound (II), more preferably 1 to 10 molar equivalents.
  • a base may be added to accelerate the reaction as necessary, and sodium carbonate, cesium carbonate, potassium carbonate, and the like are usually used as the base.
  • the amount of the base used is 1 to 10 molar equivalents relative to compound (II), preferably 1 to 5 molar equivalents.
  • the metal catalyst a commercially available copper catalyst (e.g., CuI, etc.) used in coupling can be used, and it is preferable to add a catalytic amount to compound (II).
  • the reaction can be carried out by heating at 30 to 150° C. for 1 to 48 hours, preferably at 80 to 130° C.
  • Compound (I) can also be produced under Buchwald-Hartwig reaction conditions using a palladium catalyst instead of a copper catalyst.
  • Compound (I) can also be produced by a Chan-Lam-Evans coupling reaction using a compound in which the halogen X of compound R 2 -X is converted to a boronic acid or a boronic acid ester thereof.
  • compound (I) of the present invention can be obtained by subjecting compound (II) to a nucleophilic substitution reaction in the presence of 1 to 5 molar equivalents, preferably 1 to 2.5 molar equivalents of compound R 2 -X and 1 to 5 molar equivalents, preferably 1 to 3.5 molar equivalents of a base such as cesium carbonate, in a solvent.
  • the solvent may be any solvent inactive to the reaction, and is not particularly limited, but preferably DMSO or DMF can be used.
  • the reaction can be carried out by heating at 30 to 150° C. for 1 to 24 hours, and preferably at 50 to 80° C. for 1 to 7 hours.
  • Compound (I) of the present invention can also be produced by the method shown in Scheme 2, for example, using compound (III). (In the formula, R 1 , R 2 and Z are as defined above.)
  • Compound (I) of the present invention can be produced by Chan-Lam-Evans coupling reaction using compound (III) and boronic acid (IV). That is, compound (I) can be produced by reacting a mixture of compound (III) and boronic acid (IV) in a solvent in the presence of a metal catalyst such as copper or nickel, using a base and an additive as necessary.
  • the solvent used in the reaction may be any solvent inert to the reaction, and is not particularly limited, but preferably dichloromethane or dichloroethane can be used.
  • the reaction can be carried out by heating at 30 to 150° C. for 1 to 24 hours, preferably at 50 to 80° C. for 1 to 7 hours.
  • the boronyl group of compound (IV) can be replaced with a boronic acid ester group for use in the reaction.
  • Compound (I) can also be produced by subjecting compound (III) to a nucleophilic substitution reaction with a Z-introducing agent in which the boronic acid moiety of compound (IV) is a suitable leaving group, in the presence of a suitable base.
  • the compound (I) of the present invention can also be produced, for example, according to Scheme 3.
  • the compound (I) of the present invention can be produced by converting the ester group of the compound (V) to a carboxamide group by aminolysis reaction. That is, the compound (I) can be produced by reacting the compound (V) with ammonia in a solvent under heating.
  • the solvent may be any solvent inert to the reaction, and is not particularly limited, but preferably ethanol or methanol can be used.
  • ammonia an aqueous solution, an alcohol solution, etc. can be used, or ammonia gas can be directly blown into the reaction solution.
  • the reaction can be carried out by heating at 50 to 150° C. for 1 to 48 hours, preferably at 80 to 100° C. for 1 to 15 hours.
  • the conversion to the carboxamide group can also be carried out by a method commonly used in organic synthesis, namely, hydrolysis of an ester followed by an amidation reaction using a coupling agent.
  • the compound (I) of the present invention can also be produced, for example, according to Scheme 4.
  • Scheme 4 In the formula, R 1 , R 2 and Z are defined as above, and X represents a halogen.
  • Compound (I) of the present invention can be produced by a cross-coupling reaction such as Suzuki coupling reaction using compound (VI) and compound (VII) (for example, see known literature (N. Miyaura, et al., J. Am. Chem. Soc., 107, 972 (1985)., N. Miyaura, A. Suzuki, Chem. Rev. 95, 2457 (1995)) for the conditions of Suzuki coupling reaction).
  • compound (I) can be produced by reacting a mixture of compound (VI) and compound (VII) in a solvent in the presence of a metal catalyst such as palladium or nickel, and using a base and an additive as necessary.
  • a metal catalyst such as palladium or nickel
  • the solvent used in the reaction include THF, dioxane, toluene, dimethoxyethane, methanol, ethanol, and acetonitrile. It is also suitable to use two or more of these solvents in combination, or to further mix them with water.
  • a mixed solvent of dioxane and water, or a mixed solvent of toluene, methanol, and water is preferred.
  • Compound (VII) is preferably used in a molar equivalent or excess amount relative to compound (VI), more preferably 1 to 10 molar equivalents.
  • a base may be added to accelerate the reaction, and sodium carbonate, cesium carbonate, potassium carbonate, cesium fluoride, and the like are usually used as the base.
  • the amount of the base used is 1 to 10 molar equivalents relative to compound (VI), preferably 1 to 5 molar equivalents.
  • a commercially available palladium catalyst e.g., PdCl2 (Amphos), PdCl2 (dppf), Pd2 (dba) 3 , Pd( PPh3 ) 4 , etc.
  • a catalytic amount i.e., 0.1 to 0.5 molar equivalents
  • the boronyl group of compound (VII) can be replaced with a boronic acid ester group for use in the reaction.
  • Compound (VII) which is one of the raw materials in Scheme 4, is a commercially available product or can be obtained by a known method or a method analogous thereto.
  • the compound (Ia) of the present invention in which R 1 in formula (I) is an amino group which may have a substituent, can also be produced according to scheme 5, for example.
  • R 2 , R 13 , R 14 and Z are the same as defined above.
  • Compound (I-a) can be produced by condensation reaction of the carboxyl group of compound (VIII) with amine (IX). That is, compound (VIII) can be obtained by reacting 0.9 to 5 molar equivalents, preferably 1 to 2 molar equivalents, of amine (IX) in a solvent in the presence of an amide condensing agent commonly used in organic chemistry.
  • the solvent may be any solvent inert to the reaction, and is not particularly limited, but for example, DMF, THF, etc. can be used.
  • the reaction can be carried out by stirring at 0 to 150°C for 1 to 48 hours, but preferably by reacting at room temperature to 80°C for 1 hour to overnight.
  • the amine (IX) which is one of the raw materials in Scheme 5, is a commercially available product or can be obtained by a known method or a method similar thereto.
  • compound (I-b) of the present invention having an "amino group which may have a substituent" as a substituent can also be produced using compound (I-c) of the present invention in which R 1 is represented by an alkyl group which may have a substituent and has a halogen as a substituent, for example, according to Scheme 6.
  • R 2 , Z, R 11 and R 12 are the same as defined above, and X represents a halogen.
  • Compound (I-b) can be produced by reacting compound (I-c) with amine (IX).
  • compound (I-c) can be reacted with 0.9 to 5 molar equivalents, preferably 1 to 2 molar equivalents, of amine (IX) in a solvent in the presence of a base.
  • the solvent may be any solvent inert to the reaction, and is not particularly limited.
  • DMF, THF, etc. can be used.
  • the reaction can be carried out at 0 to 150° C. with stirring for 1 to 48 hours, preferably at room temperature to 80° C. for 1 hour to overnight.
  • Compound (II) used as the starting material in Scheme 1 can be produced, for example, by the method shown in Scheme 7.
  • Scheme 7 In the formula, R1 and Z are defined as above, X represents a halogen, and PG represents a protecting group.
  • Compound (II) can be produced by deprotecting the product obtained by a cross-coupling reaction such as Suzuki coupling reaction using compound (X) and compound (VII). That is, a protected form of compound (II) can be obtained by reacting compound (X) and compound (VII) under the same conditions as in Scheme 4.
  • the deprotection reaction of the obtained protected form of compound (II) can be carried out by subjecting it to deprotection conditions generally used in organic chemistry that are suitable for the protecting group used.
  • the boronyl group of compound (VII) can also be replaced with a boronic acid ester group and used in the reaction.
  • Compound (X) used as the starting material in Scheme 7 can be produced, for example, by the method shown in Scheme 8.
  • Z is defined as above, X represents a halogen, R3 represents a lower alkyl group, and PG represents a protecting group.
  • Compound (X) can be produced by converting the ester group of compound (XI) to a carboxamide group by aminolysis reaction. The reaction can be carried out under the same conditions as in Scheme 3. The conversion to the carboxamide group can also be carried out by a method used in ordinary organic synthesis, that is, by hydrolysis of the ester followed by an amidation reaction using a coupling agent.
  • Compound (XI) used as the starting material in Scheme 8 can be produced, for example, by the method shown in Scheme 9.
  • Z is as defined above, X represents a halogen, R3 represents a lower alkyl group, and PG represents a protecting group.
  • Compound (XI) can be produced by treating compound (XII) with a halogenating agent typically used in organic chemistry, such as N-bromosuccinimide (NBS) or N-chlorosuccinimide (NCS), etc. That is, compound (XI) can be obtained by reacting compound (XII) with 0.5 to 2 molar equivalents, preferably 0.8 to 1.1 molar equivalents, of NBS or NCS in a solvent.
  • NBS N-bromosuccinimide
  • NCS N-chlorosuccinimide
  • the solvent may be any solvent inert to the reaction, and is not particularly limited, but preferably dichloromethane or DMF can be used.
  • the reaction can be carried out by stirring at -20 to 30°C for 0.25 to 10 hours, preferably at -10 to 10°C for 0.5 to 5 hours.
  • Compound (XII) used as the starting material in Scheme 9 can be produced, for example, by the method shown in Scheme 10.
  • Z is defined as above, R3 represents a lower alkyl group, and PG represents a protecting group.
  • Compound (XII) can be produced by Chan-Lam-Evans coupling reaction using compound (XIII) and boronic acid (IV). The reaction conditions can be the same as those in Scheme 2. Furthermore, the boronyl group of compound (IV) can be replaced with a boronic acid ester group for use in the reaction.
  • Compound (XII) can also be produced by subjecting compound (XIII) to a nucleophilic substitution reaction with a Z-introducing agent in which the boronic acid moiety of compound (IV) is a suitable leaving group, in the presence of a suitable base.
  • Compound (XIII) used as the starting material in Scheme 10 can be produced, for example, by the method shown in Scheme 11. (In the formula, R3 represents a lower alkyl group, and PG represents a protecting group.) Compound (XIII) can be produced by reacting aminopyrazole (XIV) with thiophosgene, treating with aqueous ammonia to obtain thioamide (XV), and reacting the thioamide (XV) with ethyl bromopyruvate to form a thiazole ring.
  • aminopyrazole (XIV) is reacted with 0.5 to 2 molar equivalents, preferably 0.9 to 1.5 molar equivalents of thiophosgene, if necessary, in the presence of a base, and then an aqueous ammonia solution is added to obtain thioamide (XV).
  • Compound (XIII) can be obtained by reacting this thioamide (XV) with 0.5 to 2 molar equivalents, preferably 0.8 to 1.5 molar equivalents of ethyl bromopyruvate in a solvent such as ethanol in the presence of a base such as potassium carbonate.
  • the aminopyrazole (XIV) used as a starting material in Scheme 11 is either commercially available or can be easily prepared using a method commonly used in organic synthesis using 4-nitropyrazole or the like as a starting material.
  • Compound (III) used as a starting material in Scheme 2 can be produced, for example, by the method shown in Scheme 12.
  • R 1 and R 2 are defined as above, and X represents a halogen.
  • Compound (III) can be produced by a cross-coupling reaction such as Suzuki coupling reaction using compound (XVI) and compound (VII).
  • the cross-coupling reaction can be carried out under the same conditions as in Scheme 4.
  • the boronyl group of compound (VII) can also be replaced with a boronic acid ester group for use in the reaction.
  • Compound (XVI) used as the starting material in Scheme 12 can be produced, for example, by the method shown in Scheme 13. (wherein R2 is as defined above, R3 represents a lower alkyl group, and X represents a halogen.)
  • Compound (XVI) can be produced by converting the ester group of compound (XVII) to a carboxamide group by aminolysis reaction. The reaction can be carried out under the same conditions as in Scheme 3. The conversion to the carboxamide group can also be carried out by a method used in ordinary organic synthesis, that is, hydrolysis of the ester followed by an amidation reaction using a coupling agent.
  • Compound (XVII) used as the starting material in Scheme 13 can be produced, for example, by the method shown in Scheme 14. (wherein R2 is as defined above, R3 represents a lower alkyl group, and X represents a halogen.)
  • Compound (XVII) can be produced by treating compound (XVIII) with a halogenating agent commonly used in organic chemistry. The reaction conditions can be the same as those in Scheme 9.
  • Compound (XVIII) used as a starting material in Scheme 14 can be produced, for example, in Scheme 10, by using an aminopyrazole derivative having an R 2 group instead of the protecting group of compound (XIII) as a starting material.
  • Compound (V) used as a starting material in Scheme 3 can be produced, for example, by the method shown in Scheme 15.
  • R 1 , R 2 and Z are defined as above, R 3 represents a lower alkyl group, PG represents a protecting group, and X represents a halogen.
  • R 2 is an optionally substituted aryl group or an optionally substituted heteroaryl group
  • compound (V) can be produced by deprotecting the protecting group of compound (XIX) and then subjecting the resulting compound to a coupling reaction such as Ullmann condensation reaction or Buchwald-Hartwig reaction with compound R 2 -X.
  • the reaction conditions can be the same as those in Scheme 1.
  • Compound (V) can also be produced by a Chan-Lam-Evans coupling reaction using a compound in which the halogen X of compound R 2 -X is converted to a boronic acid or a boronic acid ester thereof.
  • R 2 is an alkynyl group which may have a substituent
  • compound (V) can be produced by deprotecting the protecting group of compound (XIX) and then subjecting it to a nucleophilic substitution reaction with compound R 2 -X.
  • the reaction can be carried out under the same reaction conditions as in Scheme 1.
  • Compound (XIX) used as the starting material in Scheme 15 can be produced, for example, by the method shown in Scheme 16.
  • R1 and Z are defined as above, R3 represents a lower alkyl group, PG represents a protecting group, and X represents a halogen.
  • Compound (XIX) can be produced by a cross-coupling reaction such as Suzuki coupling reaction using compound (XI) and compound (VII).
  • the cross-coupling reaction can be carried out under the same conditions as in Scheme 4.
  • the boronyl group of compound (VII) can also be replaced with a boronic acid ester group for use in the reaction.
  • Compound (VI) used as the starting material in Scheme 4 can be produced, for example, by the method shown in Scheme 17.
  • R2 and Z are defined as above, PG represents a protecting group, and X represents a halogen.
  • compound (VI) can be produced by deprotecting the protecting group of compound (X) and then subjecting the compound to a coupling reaction such as Ullmann condensation reaction or Buchwald-Hartwig reaction with compound R 2 -X.
  • the reaction conditions can be the same as those in Scheme 1.
  • Compound (VI) can also be produced by a Chan-Lam-Evans coupling reaction using a compound in which the halogen X of compound R 2 -X is converted into a boronic acid or a boronic acid ester thereof.
  • R 2 is an alkynyl group which may have a substituent
  • compound (VI) can be produced by deprotecting the protecting group of compound (X) and then subjecting the compound to a nucleophilic substitution reaction with compound R 2 -X.
  • the reaction can be carried out under the same reaction conditions as in Scheme 1.
  • Compound (VIII) used as the starting material in Scheme 5 can be produced, for example, by the method shown in Scheme 18.
  • R2 and Z are as defined above, and R6 represents lower alkyl.
  • Compound (VIII) can be produced by hydrolyzing the ester group of compound (XX).
  • the hydrolysis reaction can be carried out under reaction conditions generally used in organic chemistry, and alkaline conditions (sodium hydroxide, lithium hydroxide, etc.) or acidic conditions (hydrochloric acid, sulfuric acid, etc.) can be used.
  • Compound (XX) used as the starting material in Scheme 18 can be produced, for example, by the method shown in Scheme 19.
  • R2 and Z are as defined above, R6 represents lower alkyl, and X represents halogen.
  • compound (XX) can be produced by deprotecting the protecting group of compound (XXI) and then subjecting the compound to a coupling reaction such as Ullmann condensation reaction or Buchwald-Hartwig reaction with compound R 2 -X. The reaction can be carried out under the same reaction conditions as in Scheme 1.
  • Compound (XX) can also be produced by a Chan-Lam-Evans coupling reaction using a compound in which the halogen X of compound R 2 -X is converted to a boronic acid or a boronic acid ester thereof.
  • R2 is an alkynyl group which may have a substituent
  • compound (XX) can be produced by deprotecting the protecting group of compound (XXI) and then subjecting the compound to a nucleophilic substitution reaction with compound R2 -X. The reaction can be carried out under the same reaction conditions as in Scheme 1.
  • Compound (XXI) used as the starting material in Scheme 19 can be produced, for example, by the method shown in Scheme 20.
  • Z is as defined above, R6 represents a lower alkyl group, PG represents a protecting group, and X represents a halogen.
  • Compound (XXI) can be produced by a cross-coupling reaction such as Suzuki coupling reaction using compound (X) and compound (XXII). The cross-coupling reaction can be carried out under the same conditions as in Scheme 4.
  • the boronyl group of compound (XXII) can be replaced with a boronic acid ester group and used in the reaction.
  • the compound (Ic) of the present invention used as the starting material in Scheme 6 can be produced from the compound (Id) of the present invention, which is represented by formula (I) in which R 1 is a methyl group, by, for example, the method shown in Scheme 21. (In the formula, R2 and Z are defined as above, and X represents a halogen.)
  • Compound (I-c) can be produced by treating compound (I-d) with a halogenating agent used in ordinary organic chemistry, such as benzyltrimethylammonium dichloroiodide, NBS, NCS, etc.
  • compound (I-c) can be obtained by reacting compound (I-d) with 0.5 to 5.0 molar equivalents, preferably 0.8 to 2.0 molar equivalents of a halogenating agent in a solvent.
  • the solvent may be any one inert to the reaction, and is not particularly limited, and dichloromethane, THF, DMF, etc. can be used.
  • the reaction can be carried out by stirring at -20 to 100°C for 0.25 to 10 hours, but preferably at room temperature to 80°C for 0.5 to 5 hours.
  • the compound (I) of the present invention or a pharma- ceutically acceptable salt thereof can be prepared in the form of a conventional pharmaceutical preparation (pharmaceutical composition) suitable for oral, parenteral or topical administration.
  • a conventional pharmaceutical preparation pharmaceutical composition
  • Preparations for oral administration include solid preparations such as tablets, granules, powders, capsules, and liquid preparations such as syrups. These preparations can be prepared by conventional methods.
  • Solid preparations can be prepared by using conventional pharmaceutical carriers such as lactose, starch such as corn starch, crystalline cellulose such as microcrystalline cellulose, hydroxypropyl cellulose, calcium carboxymethyl cellulose, talc, magnesium stearate, etc.
  • Capsules can be prepared by encapsulating the granules or powders thus prepared.
  • Syrups can be prepared by dissolving or suspending the compound (I) of the present invention or a pharma- ceutically acceptable salt thereof in an aqueous solution containing sucrose, carboxymethyl cellulose, etc.
  • Preparations for parenteral administration include injections such as drip infusions.
  • Injection preparations can also be prepared by conventional methods, and can be appropriately incorporated into isotonic agents (e.g., mannitol, sodium chloride, glucose, sorbitol, glycerol, xylitol, fructose, maltose, mannose), stabilizers (e.g., sodium sulfite, albumin), and preservatives (e.g., benzyl alcohol, methyl p-oxybenzoate).
  • isotonic agents e.g., mannitol, sodium chloride, glucose, sorbitol, glycerol, xylitol, fructose, maltose, mannose
  • stabilizers e.g., sodium sulfite, albumin
  • preservatives e.g., benzyl alcohol, methyl p-oxybenzoate
  • the dose of the compound (I) of the present invention or a pharma- ceutically acceptable salt thereof can be varied according to the severity of the disease, the age and body weight of the patient, the dosage form, etc., but is usually in the range of 1 mg to 1,000 mg per day for an adult, which can be administered orally or parenterally in a single dose, or in two or three divided doses.
  • the compound (I) of the present invention or a pharma- ceutical acceptable salt thereof can be used as an ALK5 inhibitor and as a reagent for experiments and research.
  • the compound (I) of the present invention that is radiolabeled can also be used as a molecular probe for PET.
  • reaction mixture was diluted with water and extracted with ethyl acetate.
  • organic layer was dried over anhydrous sodium sulfate, the solvent was removed under reduced pressure, and the residue was purified using a HPLC preparative chromatography system to obtain the title compound (21 mg).
  • reaction mixture was diluted with water and extracted with ethyl acetate.
  • organic layer was dried over anhydrous sodium sulfate, the solvent was removed under reduced pressure, and the residue was purified using a HPLC preparative chromatography system to obtain the title compound (21.3 mg).
  • Examples 9 to 183 The following example compounds [Table 1] were produced using the corresponding raw materials (commercially available products, or compounds derived from commercially available compounds by known methods or methods similar thereto) according to the methods described in the above examples, and, if necessary, by appropriately combining methods commonly used in organic synthetic chemistry. The physicochemical data of each compound is shown in [Table 2].
  • Step 8 Preparation of 2-[(1-(but-2-yn-1-yl)-1H-pyrazol-4-yl)(cyclopropylmethyl)amino)-5-[4-(dimethylcarbamoyl)-1H-pyrrol-2-yl]thiazole-4-carboxamide Potassium carbonate (103 mg, 0.75 mmol) was added to a solution of 2-[(cyclopropylmethyl)(1H-pyrazol-4-yl)amino]-5-[4-(dimethylcarbamoyl)-1H-pyrrol-2-yl]thiazole-4-carboxamide (150 mg, 0.38 mmol) and 1-bromo-2-butyne (103 mg, 0.75 mmol) in DMF (3 mL), and the mixture was reacted at 120° C.
  • Examples 185 to 226 The following example compounds [Table 3] were produced using the corresponding raw materials (commercially available products, or compounds derived from commercially available compounds by known methods or methods similar thereto) according to the methods described in the above examples, and, if necessary, by appropriately combining methods commonly used in organic synthetic chemistry. The physicochemical data of each compound is shown in [Table 4].
  • TR-FRET time-resolved fluorescence resonance energy transfer
  • a 10 mM DMSO solution of the test compound was further diluted with DMSO to 10 concentrations (0.00003 mM, 0.0001 mM, 0.0003 mM, 0.001 mM, 0.003 mM, 0.01 mM, 0.03 mM, 0.1 mM, 0.3 mM, 1 mM), and each was diluted 25-fold with assay buffer to prepare a drug solution (4% DMSO solution).
  • Drug or control solution (5 ⁇ L), substrate mixture (5 ⁇ L), and enzyme solution (10 ⁇ L) were mixed in wells of a 384-well black plate and reacted at room temperature for 1 hour.
  • the TR-FRET signal was measured using a multimode plate reader (EnVision, PerkinElmer) according to the protocol attached to the kit. (Method of evaluating inhibitory activity) As a blank, an assay buffer was added instead of the enzyme solution to measure. The inhibition rate (%) of the test compound was calculated according to the following formula.
  • Inhibition rate (%) (1 - (C - A) / (B - A)) x 100
  • A, B, and C respectively indicate the TR-FRET signal of a blank well, the TR-FRET signal of a control solution well, and the TR-FRET signal of a compound-added well.
  • the IC 50 value was calculated by regression analysis of the inhibition rate and the test compound concentration (logarithm).
  • the inhibitory activity against ALK5 of representative compounds of the present invention is shown in Table 5.
  • IC50 values of less than 0.05 ⁇ M are indicated by ***, 0.05 ⁇ M or more and less than 0.5 ⁇ M are indicated by **, and 0.5 ⁇ M or more and less than 5 ⁇ M are indicated by *.
  • Test Example 2 Inhibition test of Smad3 phosphorylation by intracellular ALK5 (culture of cells used) A549 cells (ATCC No. CCL-185) were cultured in a T75 flask at 37°C in a 5% CO2 incubator using D-MEM medium (Nacalai, #08459-64) (hereinafter referred to as growth medium) supplemented with 10% fetal bovine serum (hereinafter referred to as FBS) (AusGene) and 1% penicillin-streptomycin (Nacalai).
  • D-MEM medium Nacalai, #08459-644
  • FBS fetal bovine serum
  • penicillin-streptomycin Nacalai
  • the cultured A549 cells were diluted with growth medium to a cell density of 3.3 x 105 cells/mL, and the resulting cell suspension was seeded in a 24-well culture plate (Falcon, 353226) at 3.3 x 105 cells/mL/well, and cultured overnight at 37°C in a 5% CO2 incubator. The next day, the growth medium was removed with an aspirator and immediately washed with FBS-free D-MEM medium (hereinafter referred to as medium) kept at 37°C. The cell plate was prepared by adding 0.989 mL of medium to each well.
  • TGF ⁇ 1 (Peprotec, PEP-100-21-10) adjusted to 200 ng/mL in culture medium was added to each well of the cell plate (10 ⁇ L of culture medium alone was added to the control well), and the plate was further incubated for 30 minutes at 37° C. in a 5% CO 2 incubator.
  • the mixture was mixed with SDS-sample buffer and reacted at 95°C for 5 minutes to denature the proteins, and a sample solution was prepared. 5 ⁇ g/12 ⁇ L of the sample solution was applied to each well of a 5-20% gradient acrylamide gel (Nacalai, No. 13064-04) and electrophoresis was performed. The proteins in the gel were then transferred to a PVDF membrane using a Transblot Turbo transfer system (Bio-Rad). (Detection of phosphorylated Smad3) The transferred PVDF membrane was subjected to blocking treatment with 2% ECL prime blocking agent (GE Healthcare), and then reacted overnight at 4° C.
  • ECL prime blocking agent GE Healthcare
  • the combinations and dilution concentrations of the primary and secondary antibodies used in this test are as follows. (Method of evaluating inhibitory activity) The detected bands were quantified by densitometry (ImageQuant TL analysis software), and the inhibition rate was calculated from the band intensity in each group, taking the luminescence of the phosphorylated Smad3 band in the compound-free and TGF ⁇ -stimulated group as 100% and the luminescence of the phosphorylated Smad3 band in the compound-free and TGF ⁇ -unstimulated group as 0%. Each phosphorylated Smad3 band was corrected with ⁇ -actin. The inhibition rate (%) of Smad3 phosphorylation by the test compound was calculated according to the following formula.
  • Inhibition rate (%) (1 - (C - A) / (B - A)) x 100
  • A, B, and C respectively show the luminescence of the phosphorylated Smad3 band in the group without addition of compound and without stimulation with TGF ⁇ , the group without addition of compound and stimulation with TGF ⁇ , and the group with addition of compound and stimulation with TGF ⁇ .
  • the IC 50 value was calculated by regression analysis of the inhibition rate and the test compound concentration (logarithm).
  • the inhibitory activity of intracellular Smad3 phosphorylation of representative compounds of the present invention is shown in Table 7.
  • IC50 values of less than 0.1 ⁇ M are indicated by ***, those of 0.1 ⁇ M or more and less than 0.3 ⁇ M are indicated by **, and those of 0.3 ⁇ M or more are indicated by *.
  • Test Example 3 Inhibition test of regulatory T cell (Treg) differentiation induction by TGF- ⁇ stimulation
  • Spleens were collected from mice (BALB/cCrSlc, female, 7 weeks old), and mouse lymphocytes were separated from a suspension of splenocytes (FBS-free IMDM medium) prepared using a cell strainer using HISTOPAQUE-1083 (Sigma).
  • CD4 positive T cells were isolated from these mouse lymphocytes using EasySep Mouse Naive CD4+ T Cell Isolation Kit (manufactured by STEMCELL technologies) according to the protocol attached to the kit.
  • a compound solution prepared by diluting a DMSO solution of a test compound 100-fold with IMDM medium supplemented with IL-2-containing FBS was added to each well.
  • CD4-positive T cells were suspended in IMDM medium supplemented with IL-2-containing FBS (2 ⁇ 10 5 cells/mL), 0.1 mL of the solution was added to each well, and the mixture was incubated for 1 hour in a CO 2 incubator.
  • the cells were collected from each well, stained with FITC anti-mouse CD4 Antibody (BioLegend), APC anti-mouse CD25 Antibody (BioLegend), and FOXP3 Monoclonal Antibody (FJK-16s) PE (eBioscience), and the proportion of the Treg fraction (CD4+/CD25+/Foxp3+) was measured using a flow cytometer.
  • IMDM medium supplemented with IL-2-containing FBS was used instead of TGF- ⁇ 1.
  • the IC 50 value was calculated by regression analysis of the change in the proportion of the Treg fraction and the test compound concentration (logarithm).
  • the inhibitory activity of representative compounds of the present invention in inducing Treg differentiation by TGF- ⁇ stimulation is shown in Table 8.
  • IC50 values of less than 0.1 ⁇ M are indicated by ***, 0.1 ⁇ M or more and less than 0.5 ⁇ M by **, and 0.5 ⁇ M or more by *.
  • Table 8 In this test, as shown in Table 8, compound (I) of the present invention inhibited TGF- ⁇ signaling in naive T cells and strongly suppressed the induction of differentiation into Treg by TGF- ⁇ stimulation.
  • the results of Test Example 3 show that compound (I) of the present invention inhibits intracellular TGF- ⁇ signaling and has a strong suppressive effect on the induction of Treg differentiation.
  • Test Example 4 Effect of Combination with Anti-PD-1 Antibody in an Allograft Mouse Model of Mouse Colon Cancer Cell Line CT26.WT
  • the effect of combination with an immune checkpoint inhibitor in cancer immunotherapy was examined using a syngeneic mouse tumor model (subcutaneous transplant) of the mouse colon cancer cell line CT26.WT.
  • CT26.WT cells were adjusted to a cell density of 1 x 107 cells/mL in HBSS(-) medium (manufactured by Nacalai) to prepare a cell preparation for transplantation.
  • 0.1 mL of this cell preparation for transplantation was subcutaneously transplanted into the back of a BALB/cCrslc mouse (female, 7 weeks old, Japan SLC).
  • mice Four days after the cancer cells were transplanted, the mice were divided into groups so that the average tumor volume (see the calculation formula below) of the cancer-bearing mice was close to each other.
  • Anti-PD-1 antibody Bio X Cell, clone RMP1-14; catalog No.
  • BE0146 was prepared with physiological saline to a concentration of 1 mg/mL immediately before administration.
  • Each mouse (6 mice per group) transplanted with cancer cells was forced to orally administer 0.1 mL of the test substance (100 mg/kg) or the solvent per 10 g of body weight on the day once a day from the 4th to the 20th day after transplantation. Drugs were suspended on the 9th, 10th, 16th, and 17th days after transplantation.
  • the antibody administration group and the drug combination group were intraperitoneally administered 0.1 mL of the antibody solution (10 mg/kg) per 10 g of body weight on the day twice a week (5 times in total), and saline was administered intraperitoneally to the other groups.
  • FIG. 1 shows the change in tumor volume over time in each group.
  • the compounds of the present invention, Example 6 and Example 7, showed a significant tumor growth suppression/tumor regression effect when used in combination with an anti-PD-1 antibody. This confirmed that the compounds of the present invention showed an excellent antitumor effect when used in combination with an immune checkpoint inhibitor, and are useful in the treatment of cancer.
  • Test Example 5 Combination effect of anti-PD-1 antibody in allograft mouse model of mouse colon cancer cell line CT26.WT 2 (Preparation of tumor-bearing model) CT26.WT cells were adjusted to a cell density of 1 x 107 cells/mL with HBSS(-) medium to prepare a cell preparation for transplantation. 0.1 mL of this cell preparation for transplantation was subcutaneously transplanted into the back of a BALB/cCrslc mouse (female, 7 weeks old, Japan SLC). On the third day after the cancer cells were transplanted, the mice were divided into groups so that the average tumor volume (see the calculation formula in Test Example 4) of the cancer-bearing mice was close to each other.
  • test substance preparation of sample solution for administration of test substance
  • Each mouse transplanted with cancer cells (10 mice in the solvent group, 9 mice in the compound group of Example 184, 9 mice in the antibody PD-1 antibody group, and 8 mice in the compound of Example 184 and antibody PD-1 antibody combination group) was forcibly administered 0.1 mL of the test substance (30 mg/kg) or the solvent per 10 g of body weight on the day once a day from the 3rd to the 21st day after transplantation. Drugs were suspended on the 8th, 9th, 15th, and 16th days after transplantation.
  • the antibody administration group and the drug combination group were intraperitoneally administered 0.1 mL of antibody solution (10 mg/kg) per 10 g of body weight on the day twice a week (6 times in total), and saline was intraperitoneally administered to the other groups.
  • the mice were observed until the 21st day, and the tumor diameter was measured several times a week to calculate the tumor volume of each mouse using the formula in Test Example 4 to evaluate the antitumor effect.
  • Figure 2 shows the change in tumor volume over time in each group.
  • the compound of the present invention, Example 184 showed a significant tumor growth inhibitory and tumor regression effect when used alone or in combination with an anti-PD-1 antibody. This confirmed that the compound of the present invention showed an excellent antitumor effect when used in combination with an immune checkpoint inhibitor, and is useful in the treatment of cancer.
  • the compounds provided by the present invention are useful for treating diseases known to be associated with abnormal cell responses via the TGF ⁇ signal pathway, particularly cancer, and are also useful for preventing tumor metastasis and recurrence by targeting cancer stem cells. Furthermore, when used in combination with immune checkpoint inhibitors, they are useful for expanding and enhancing the therapeutic effect in cancer immunotherapy. They are also useful for treating and preventing fibrotic diseases, etc. Furthermore, they are useful as experimental and research reagents as ALK5 inhibitors and TGF ⁇ signal inhibitors.

Landscapes

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

Abstract

La présente invention concerne un dérivé de thiazole qui a une action inhibitrice d'ALK 5 et qui est représenté par la formule (1) (voir la description pour R1, R2, et Z dans la formule), ou un sel pharmaceutiquement acceptable de celui-ci.
PCT/JP2023/041996 2022-11-25 2023-11-22 Nouveau dérivé de thiazole WO2024111626A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
JP2022-188246 2022-11-25
JP2022188246 2022-11-25
JP2023-104088 2023-06-26
JP2023104088 2023-06-26

Publications (1)

Publication Number Publication Date
WO2024111626A1 true WO2024111626A1 (fr) 2024-05-30

Family

ID=91196177

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2023/041996 WO2024111626A1 (fr) 2022-11-25 2023-11-22 Nouveau dérivé de thiazole

Country Status (1)

Country Link
WO (1) WO2024111626A1 (fr)

Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2005527590A (ja) * 2002-04-04 2005-09-15 バイオジェン， インコーポレイテッド 三置換ヘテロアリールおよびそれらを製造し使用する方法
JP2006516603A (ja) * 2003-01-27 2006-07-06 ファイザー・プロダクツ・インク イソチアゾール誘導体
JP2006517592A (ja) * 2003-02-12 2006-07-27 バイオジェン・アイデック・エムエイ・インコーポレイテッド ピラゾールおよびそれらを製造し使用する方法
JP2008511631A (ja) * 2004-08-31 2008-04-17 バイオジェン・アイデック・エムエイ・インコーポレイテッド ＴＧＦ−βインヒビターとしてのピリミジニルイミダゾール
JP2013507932A (ja) * 2009-10-16 2013-03-07 ザ スクリプス リサーチ インスティチュート 多能性細胞の誘導法
JP2015517453A (ja) * 2012-05-24 2015-06-22 カルナバイオサイエンス株式会社 新規二環式チアゾール化合物
JP2017537886A (ja) * 2014-10-22 2017-12-21 ダナ−ファーバー キャンサー インスティテュート， インコーポレイテッド 増殖性疾患を処置するためのチアゾリル含有化合物
WO2020012357A1 (fr) * 2018-07-10 2020-01-16 Integral Biosciences Pvt. Ltd. Composés hétérocycliques et procédés d'utilisation
WO2020103817A1 (fr) * 2018-11-20 2020-05-28 南京圣和药业股份有限公司 INHIBITEUR DE TGF-βR1 ET SON UTILISATION
WO2020146819A1 (fr) * 2019-01-10 2020-07-16 Tolero Pharmaceuticals, Inc. Inhibiteurs d'alk5 pour le traitement du syndrome myélodysplasique

Patent Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2005527590A (ja) * 2002-04-04 2005-09-15 バイオジェン， インコーポレイテッド 三置換ヘテロアリールおよびそれらを製造し使用する方法
JP2006516603A (ja) * 2003-01-27 2006-07-06 ファイザー・プロダクツ・インク イソチアゾール誘導体
JP2006517592A (ja) * 2003-02-12 2006-07-27 バイオジェン・アイデック・エムエイ・インコーポレイテッド ピラゾールおよびそれらを製造し使用する方法
JP2008511631A (ja) * 2004-08-31 2008-04-17 バイオジェン・アイデック・エムエイ・インコーポレイテッド ＴＧＦ−βインヒビターとしてのピリミジニルイミダゾール
JP2013507932A (ja) * 2009-10-16 2013-03-07 ザ スクリプス リサーチ インスティチュート 多能性細胞の誘導法
JP2015517453A (ja) * 2012-05-24 2015-06-22 カルナバイオサイエンス株式会社 新規二環式チアゾール化合物
JP2017537886A (ja) * 2014-10-22 2017-12-21 ダナ−ファーバー キャンサー インスティテュート， インコーポレイテッド 増殖性疾患を処置するためのチアゾリル含有化合物
WO2020012357A1 (fr) * 2018-07-10 2020-01-16 Integral Biosciences Pvt. Ltd. Composés hétérocycliques et procédés d'utilisation
WO2020103817A1 (fr) * 2018-11-20 2020-05-28 南京圣和药业股份有限公司 INHIBITEUR DE TGF-βR1 ET SON UTILISATION
WO2020146819A1 (fr) * 2019-01-10 2020-07-16 Tolero Pharmaceuticals, Inc. Inhibiteurs d'alk5 pour le traitement du syndrome myélodysplasique

Similar Documents

Publication Publication Date Title
JP6911031B2 (ja) 免疫調節剤としての複素環化合物
CN107709333B (zh) 羟基酯衍生物、其制备方法以及含有它们的药物组合物
KR101816942B1 (ko) 키누레닌 생성 억제 작용을 갖는 함질소 복소환 화합물
CN107428758B (zh) 丙烯酸类衍生物、其制备方法及其在医药上的用途
TW201910306A (zh) Nlrp3發炎體之選擇性抑制劑
EP3889134A1 (fr) Composé inhibant la transduction du signal pge2/ep4, son procédé de préparation et ses applications thérapeutiques
WO2020015716A1 (fr) Immunomodulateurs, compositions et procédés associés
JP2022542669A (ja) Ｉｒａｋ４キナーゼ阻害剤及びその調製方法
AU2019251096B2 (en) Pladienolide compounds and their use
JPWO2016031987A1 (ja) オートタキシン阻害活性を有するピリミジノン誘導体
WO2020038460A1 (fr) Nouvel inhibiteur de dérivé de quinoléine
CA3056302A1 (fr) Composes chimiques
JP2021525810A (ja) トリプトファン異化のモジュレーター
CN113166148B (zh) 作为cdk-hdac双通路抑制剂的杂环化合物
WO2024111626A1 (fr) Nouveau dérivé de thiazole
WO2020020374A1 (fr) Composé de benzène polysubstitué et son procédé de préparation et son utilisation
CN111662227A (zh) 邻氨基吡啶炔类化合物及其制备方法和用途
KR20240014050A (ko) Pd1/pd-l1 억제제로서의 화합물 및 이의 방법
KR101718299B1 (ko) 아자인돌 유도체
AU2018337138B2 (en) 2-substituted pyrazole amino-4-substituted amino-5-pyrimidine formamide compound, composition, and application thereof
WO2017070135A1 (fr) Promédicaments de dérivés 2-(4-(3-((4-amino-7-cyano-imidazo[2,1-f][1,2,4]triazin-2-yl)amino)phényl)pipérazin-1-yl)propanamide utilisés comme inhibiteurs de ck2 pour le traitement du cancer
CA3100095C (fr) Compose d'indoline-1-formamide, son procede de preparation et son utilisation medicale
CN117105919A (zh) 一种靶向抗肿瘤药物及其在制备药物中的用途
CN115960117A (zh) 含硫并环类衍生物抑制剂、其制备方法和应用
CN117730080A (zh) 吡啶并嘧啶酮类衍生物及其制备方法和用途