WO2025170440A1 - Novel nitrogen-containing heterocyclic derivative and use thereof - Google Patents

Abstract

The present invention relates to a novel nitrogen-containing heterocyclic derivative compound and a use thereof. More specifically, the present invention relates to: a novel nitrogen-containing heterocyclic derivative having an effect of inhibiting the activity of eIF2α activity kinase including GCN2; a pharmaceutically acceptable salt, optical isomer, hydrate, or solvate thereof; a pharmaceutical composition comprising the compounds as active ingredients; and a use thereof.

Description

Novel nitrogen-containing heterocyclic derivatives and uses thereof

The present invention relates to a novel nitrogen-containing heterocyclic derivative compound and its use, and more particularly, to a novel nitrogen-containing heterocyclic derivative having an activity inhibitory effect on eIF2α-activated kinases including GCN2, a pharmaceutically acceptable salt, optical isomer, hydrate or solvate thereof, a pharmaceutical composition comprising such a compound as an active ingredient, and its use.

Eukaryotic initiation factor-2α (eIF2α) kinase is a well-characterized serine-threonine kinase that is activated by the eIF2α activating kinase family, which includes heme-regulated inhibitor (HRI), double-stranded RNA-dependent protein kinase (PKR), PKR-like endoplasmic reticulum kinase (PERK), and general control nonderepressible 2 (GCN2). The eIF2α kinase reduces general translation in response to various cellular stresses by promoting a program of stress-induced gene expression. These kinases perform important and essential functions in response to infection, proteotoxicity, and low levels of essential nutrients such as amino acids and heme, and also play a crucial role in the induction and development of viral pathogenic cancers.

In particular, GCN2 (General Control Nonderepressible 2) is a protein kinase expressed in eukaryotes as a cellular response to amino acid starvation (Castilho et al., 2014, Biochim. Biophys. Acta 1843: 1948-1968). GCN2 is activated and phosphorylated by interacting with deacetylated tRNAs that accumulate as a result of amino acid starvation. Cellular stress factors such as UV irradiation, redox stress, or proteasome inhibition can also indirectly induce GCN2 activation (Wek et al., 2006, Biochem. Soc. Trans. 34: 7-11). GCN2 is activated under certain stresses and phosphorylates eIF2α (eukaryotic initiation factor 2α), a protein modification initiation regulator. Phosphorylation of eIF2α induces specific expression of stress-related target genes by cap-dependent initiation via the mammalian transcription factor ATF4 (Vattem and Wek, 2004, PNAS 101(31) 11269-11274).

Induction of cellular responses to amino acid deprivation has emerged as an important mechanism for regulating the mammalian immune system, particularly in certain disease settings, including cancer and autoimmunity. Various immunosuppressive cell types implicated in the control of immune responses in these settings, including endogenous dendritic cells, myeloid-derived suppressor cells (MDSCs), endogenous/M2 macrophages, and cancer cells themselves, have been reported to utilize amino acid deprivation to suppress T-cell responses (Munn et al., 2004, J Clin Invest, 114, 280-290). This is achieved by the intracellular transport of amino acids, along with the overexpression of amino acid catabolizing enzymes in these cells, such as indoleamine 2,3 dioxygenase (IDO), tryptophan 2,3 dioxygenase (TDO), and arginine catabolizing enzymes arginase 1 and 2 (ARG1 and ARG2). Consequently, these cells can reduce the extracellular concentration of specific amino acids in the area where they reside, thereby inducing GCN2 activity in nearby T cells in an antigen-specific manner (Munn et al., 2004, J Clin Invest, 114: 280-290). In vitro and in vivo mouse systems, depletion of local tryptophan or arginine concentrations, for example by IDO- or ARG1-expressing dendritic cells, has been reported to induce proliferation inhibition and anergy in T cells in a GCN2-dependent manner (Fletcher et al., 2015, Cancer Research. 75: 275-283). Furthermore, the induction and/or maintenance of MDSCs and immunosuppressive regulatory T cells (T-regs) may depend on GCN2 activity under amino acid starvation conditions. Another study implicates IDO-mediated GCN2 activation in tolerogenic macrophages as a key mechanism for suppressing systemic autoimmune responses against apoptotic cells (Ravishankar et al., 2015, Proc. Natl Acad. Sci. USA 112, 10774-10779). These findings suggest that GCN2 may be a potential key factor in the immunosuppressive effects of amino acid deficiencies associated with various disease states.

Furthermore, the accumulation of amyloid-β (Aβ), generated from amyloid precursor protein by γ-secretase within the cerebral cortex, may be an important cause of Alzheimer's disease. In autophagy-impaired cells, the regulation of γ-secretase activity is associated with GCN2-eIF2α-ATF4 signaling (Ohta et al., 2010, Autophagy. 6(3): 345-52). Since the autophagy-lysosome system has an important regulatory function for γ-secretase activity through GCN2, Aβ accumulation may be induced when autophagy is impaired, and thus GCN2 may be a therapeutic target for reducing Aβ production.

[Prior Art Literature]

[Patent Document]

International Publication No. WO2019-148136 (Published: August 1, 2019)

International Publication No. WO2019-148132 (Published: August 1, 2019)

International Publication No. WO2013-110309 (Published: August 1, 2013)

International Publication No. WO2018-030466 (Published: February 15, 2018)

The purpose of the present invention is to provide a novel nitrogen-containing heterocyclic derivative compound having an activity inhibitory effect on eIF2α-activated kinases including GCN2.

Another object of the present invention is to provide a pharmaceutical composition for preventing and/or treating cancer, immune disease or Alzheimer's disease, containing the compound as an active ingredient.

Another object of the present invention is to provide a method for treating or preventing a disease caused by abnormal activity of GCN2 by administering a therapeutically effective amount of the compound.

To achieve the above purpose, the present invention provides a nitrogen-containing heterocyclic derivative compound represented by the following chemical formula I, and a pharmaceutically acceptable salt, optical isomer, hydrate or solvate thereof.

[Chemical Formula I]

In the above chemical formula I,

Ring A is any one nitrogen-containing heterocycle selected from the group consisting of A-1 to A-5 below;

In the above A-1 to A-5,

Wavy lines ( ) indicates the attachment site;

A plurality of X ⁰ are different from each other, one of which is C and the rest are N;

A plurality of X ¹ are identical or different, and each independently is C(R) or N, or at least one is N;

A plurality of X ² are different from each other, one of which is C(R') and the rest are N;

X ³ is C(R'') or N;

R and R'' are each independently hydrogen, halogen, C ₁ -C ₃ alkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;

R' is halogen, C ₁ -C ₃ alkyl, carboxamide (-CONH ₂ ), substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;

R ³ is hydrogen, or a substituted or unsubstituted amino group;

The substituent of R ³ is a substituted or unsubstituted C ₁ -C ₆ alkyl, a substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, a substituted or unsubstituted 3 to 10 membered heterocycloalkyl, a substituted or unsubstituted C ₆ -C ₁₂ aryl, or a substituted or unsubstituted 5 to 12 membered heteroaryl;

R ⁴ to R ⁷ are each independently hydrogen, halogen, substituted or unsubstituted C ₁ -C ₃ alkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;

When each of R ⁴ to R ⁷ is alkyl, aryl or heteroaryl, their substituents may each independently be hydroxy, hydroxy C ₁ -C ₆ alkyl, or C ₁ -C ₆ alkoxy;

R ⁸ is hydrogen or halogen;

Ring B is arylene or heteroarylene substituted or unsubstituted with R ¹ ;

R ¹ is each independently hydrogen, hydroxy, cyano, nitro, (mono-, di-, or trihalogen)methyl, halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkene, C ₂ -C ₆ alkyne, C ₁ -C ₃ alkoxy, C ₃ -C ₆ cycloalkyl, or C ₁ -C ₆ dialkylamino;

L ¹ is C _0- C ₆ alkylene, C _2- C ₆ alkenylene, C _2- C ₆ alkynylene, -NR ⁹ -CO-, -CO-NR ¹⁰ -, -NR ⁹ -CO-NR ¹⁰ -, -NR ⁹ -SO ₂ -, -NR ⁹ -SO ₂ -NR ¹⁰ -, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene, cycloalkylene or heterocycloalkylene, or may be a direct bond;

L ² is -NR ⁹ -CO-, -CO-NR ¹⁰ -, -NR ⁹ -SO ₂ -, or -NR ⁹ -SO-NR ¹⁰ -;

R ⁹ and R ¹⁰ are each independently hydrogen or C ₁ -C ₆ alkyl;

Ring C is aryl or heteroaryl substituted or unsubstituted with R ² ;

R ² can be independently selected from the group consisting of hydrogen, hydroxy, hydroxy C ₁ -C ₆ alkyl, cyano, C _1- C ₆ alkyl, C _2- C ₆ alkene, C _2- C ₆ alkyne, C _1- C ₃ alkoxy, (mono-, di-, or trihalogen)methyl, halogen, C _3- C ₆ cycloalkyl, C _1- C ₆ dialkylamino, substituted or unsubstituted aryloxy or heteroaryloxy, substituted or unsubstituted arylalkyloxy or heteroarylalkyloxy, substituted or unsubstituted aryl or heteroaryl, and substituted or unsubstituted cycloalkyl or heterocycloalkyl;

p and q are 0 and 1 respectively, and p+q=1;

m is an integer from 0 to 4;

n is an integer from 0 to 5;

When R ¹ or R ² is plural, they are each the same or different.

In addition, the present invention provides a pharmaceutical composition comprising a compound of the above formula I, a pharmaceutically acceptable salt, optical isomer, hydrate or solvate thereof as an active ingredient.

In addition, the present invention provides a pharmaceutical composition having an action of inhibiting abnormal activity of eIF2α, and more specifically, provides a pharmaceutical composition having an action of inhibiting abnormal activity of one or more kinases selected from the group consisting of HRI, PKR, PERK, and GCN2.

In addition, the present invention provides a pharmaceutical composition for preventing or treating cancer, immune disease, or Alzheimer's disease caused by abnormal activity of GCN2.

In addition, the present invention provides a method for treating or preventing a disease caused by abnormal activity of GCN2, comprising administering a therapeutically effective amount of a compound of the above formula (I), a pharmaceutically acceptable salt, optical isomer, hydrate or solvate thereof, to a person in need of treatment.

The nitrogen-containing heterocyclic compound represented by chemical formula I and its derivatives provided in the present invention can be usefully used as an agent for preventing or treating immune diseases including cancer or Alzheimer's disease by inhibiting the activity of eIF2α-activated kinases including GCN2.

The definitions listed below define various terms used to describe the present invention. These definitions apply to this specification as a whole, either individually or as part of a term encompassing them, unless otherwise specified.

Additionally, unless otherwise specified, the terms to which the present invention pertains refer to a range of numbers that one of ordinary skill in the art would recognize as equivalent to (i.e., having the same function or result) the numerical values described. In many cases, these terms may include numbers rounded to the nearest significant figure. Specifically, the term "about" means about 5%, preferably about 1% to 2%, of a specified numerical value or range (e.g., a range of parts by weight, percent by weight, etc.). For example, "about 10% by weight" means about 9.5% to about 10.5% by weight, preferably about 9.8% to about 10.2% by weight.

Although exemplary methods or materials are described herein, other similar or equivalent methods are also within the scope of the present invention. All publications cited herein by reference are incorporated herein by reference in their entirety.

The term 'alkyl' as used herein, unless otherwise stated, refers to a saturated, straight-chain or branched hydrocarbon radical represented by C _n H _2n+1 , specifically a saturated, straight-chain or branched hydrocarbon radical containing 1 to 6, 1 to 8, 1 to 10, or 1 to 20 carbon atoms, respectively. Examples of these radicals include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, neopentyl, n-hexyl, heptyl, octyl radicals. For example, the term 'C ₁ -C ₆ alkyl', unless otherwise stated, refers to a straight-chain or branched hydrocarbon residue containing 1 to 6 carbon atoms. Examples include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, t-butyl, n-pentyl, n-hexyl, etc.

The term 'alkenyl' as used herein, unless otherwise stated, refers to a monovalent group derived from an unsaturated, straight-chain or branched hydrocarbon moiety having at least one carbon-carbon double bond, specifically an unsaturated, straight-chain or branched monovalent group containing between 2 and 6, between 2 and 8, between 2 and 10, or between 2 and 20 carbon atoms, respectively. Examples thereof include, but are not limited to, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, heptenyl, and octenyl radicals.

The term 'alkylene' as used herein, unless otherwise stated, includes a straight-chain or branched divalent hydrocarbon group having 1 to 15 carbon atoms, preferably 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms, and even more preferably 1 to 4 carbon atoms. Examples thereof include methylene, ethylene, trimethylene, propylene, tetramethylene, pentamethylene, and hexamethylene.

The term 'alkenylene' as used herein, unless otherwise stated, includes a straight-chain or branched divalent hydrocarbon group having 2 to 15 carbon atoms, preferably 2 to 10 carbon atoms, more preferably 2 to 6 carbon atoms, and even more preferably 2 to 4 carbon atoms, and having one or more double bonds at any position. Examples thereof include vinylene, propenylene, butenylene, and pentenylene.

The term 'haloalkyl' as used herein, unless otherwise stated, means alkyl substituted with halogen. Examples thereof include monofluoromethyl, monofluoroethyl, monofluoropropyl, 2,2,3,3,3-pentafluoropropyl, monochloromethyl, trifluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 2,2,2-trichloroethyl, 1,2-dibromoethyl, and 1,1,1-trifluoropropan-2-yl.

The term 'hydroxyalkyl' as used herein, unless otherwise stated, means a group in which one or more hydroxyl groups are substituted with hydrogen atoms bonded to carbon atoms of the 'alkyl'.

The term 'alkyloxy (or alkoxy)' as used herein, unless otherwise stated, refers to an oxygen radical having a monovalent group derived from a saturated, straight-chain or branched hydrocarbon moiety having 1 to 6, 1 to 8, 1 to 10, or 1 to 20 carbon atoms, represented by the formula OCnH2n+1. For example, 'C ₁ -C ₆ alkyloxy (or 'C ₁ -C ₆ alkoxy)', unless otherwise stated, refers to an oxygen radical having a straight-chain or branched hydrocarbon moiety having 1 to 6 carbon atoms. Examples include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, t-butoxy, pentoxy, hexoxy, and the like.

The term 'aryl' as used herein, unless otherwise stated, refers to a mono- or poly-cyclic carbocyclic ring system of 5 to 14 carbon atoms, preferably 5 to 12 carbon atoms, having one or more aromatic rings, fused or non-fused, examples of aryl include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, indenyl, andracenyl, etc.

The term 'heteroaryl' as used herein, unless otherwise stated, means a 5 to 12 membered, preferably 5 to 7 membered, monocyclic or bicyclic or higher aromatic group containing one or more heteroatoms selected from O, N and S, for example, 1 to 4, preferably 1 to 3. Examples of monocyclic heteroaryl include, but are not limited to, thiazolyl, oxazolyl, thiophenyl, furanyl, pyrrolyl, imidazolyl, isooxazolyl, pyrazolyl, triazolyl, thiadiazolyl, tetrazolyl, oxadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl and similar groups. Examples of bicyclic heteroaryls include, but are not limited to, indolyl, benzothiophenyl, benzofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzthiazolyl, benzthiadiazolyl, benztriazolyl, quinolinyl, isoquinolinyl, purinyl, furopyridinyl, and similar groups.

The term 'alkylcarbonyl' used in this specification, unless otherwise stated, means a group in which the 'alkyl' is bonded to a carbonyl group.

The term 'alkylamino' as used herein, unless otherwise stated, means a group in which the 'alkyl' is substituted with one or two hydrogen atoms bonded to the nitrogen atom of the amino group.

The term 'alkylcarbonylamino' used in this specification, unless otherwise stated, means a group in which the 'alkylcarbonyl' is substituted with one hydrogen atom that is bonded to the nitrogen atom of the amino group.

The term 'alkylcarbonylaminoalkyl' used in this specification, unless otherwise stated, means a group in which the 'alkylcarbonylamino' is bonded to the 'alkyl'.

The term 'alkylcarbonyloxy' used in this specification, unless otherwise stated, means a group in which the 'alkylcarbonyl' is bonded to an oxygen atom.

The term 'alkyloxycarbonyl' used in this specification, unless otherwise stated, means a group in which the 'alkyloxy (or alkoxy)' is bonded to a carbonyl group.

The term 'alkyloxycarbonylalkyl' used in this specification, unless otherwise stated, means a group in which the 'alkyloxycarbonyl' is bonded to the 'alkyl'.

The term 'alkyloxycarbonyloxy' used in this specification, unless otherwise stated, means a group in which the 'alkyloxycarbonyl' is bonded to an oxygen atom.

The term 'alkylsulfanyl (or alkylsulfanyl)' used in this specification, unless otherwise stated, means a group in which the 'alkyl' is substituted with a hydrogen atom bonded to a sulfur atom of sulfanyl (or sulfanyl group).

The term 'alkylsulfonyl (or alkylsulfonyl)' used herein includes a group in which the 'alkyl' is bonded to sulfonyl (or sulfonyl group), unless otherwise stated.

The term 'trialkylsilyl' as used herein, unless otherwise stated, refers to a group in which three 'alkyl' groups are bonded to a silicon atom. The three alkyl groups may be the same or different.

The term 'carbocyclic group' used herein, unless otherwise specified, means a carbocyclic group having 3 to 20 carbon atoms, preferably 3 to 16 carbon atoms, more preferably 4 to 12 carbon atoms, and includes aromatic carbocyclic groups and non-aromatic carbocyclic groups.

The term 'aromatic carbocyclic group' used in this specification includes a monocyclic or bicyclic, cyclic aromatic hydrocarbon group, unless otherwise specified.

The term 'non-aromatic carbocyclic group' used herein includes, unless otherwise specified, a monocyclic or bicyclic, cyclic saturated hydrocarbon group or a cyclic non-aromatic unsaturated hydrocarbon group. A bicyclic or bicyclic 'non-aromatic carbocyclic group' also includes a monocyclic or bicyclic or bicyclic non-aromatic carbocyclic group in which the ring in the 'aromatic carbocyclic group' is condensed.

The term 'carbon ring' used herein, unless otherwise stated, means a carbon ring having 3 to 20 carbon atoms, preferably 3 to 16 carbon atoms, more preferably 4 to 12 carbon atoms, and includes aromatic carbon rings and non-aromatic carbon rings.

The term 'aromatic carbocyclic ring' used herein includes a monocyclic or bicyclic, cyclic aromatic hydrocarbon, unless otherwise specified.

The term "non-aromatic carbocyclic ring" used herein, unless otherwise specified, includes a monocyclic or bicyclic, cyclic saturated hydrocarbon or a cyclic non-aromatic unsaturated hydrocarbon. A bicyclic or bicyclic "non-aromatic carbocyclic ring" also includes a ring in the "aromatic carbocyclic ring" condensed with a monocyclic or bicyclic or bicyclic non-aromatic carbocyclic ring.

The term 'heterocyclic group' as used herein includes aromatic heterocyclic groups and non-aromatic heterocyclic groups having one or more identical or different heteroatoms randomly selected from O, S and N in the ring, unless otherwise stated.

The term 'aromatic heterocyclic group' used herein includes, unless otherwise stated, a monocyclic or bicyclic aromatic cyclic group having one or more identical or different heteroatoms arbitrarily selected from O, S, and N within the ring. The bicyclic or bicyclic 'aromatic heterocyclic group' also includes a monocyclic or bicyclic aromatic heterocyclic group in which the ring in the above 'aromatic carbocyclic group' is condensed.

The term 'non-aromatic heterocycle' used herein includes, unless otherwise specified, a monocyclic or bicyclic, cyclic non-aromatic ring having one or more identical or different heteroatoms arbitrarily selected from oxygen atoms, sulfur atoms, and nitrogen atoms within the ring. A bicyclic or bicyclic 'non-aromatic heterocycle' also includes a monocyclic or bicyclic or bicyclic non-aromatic heterocycle in which each ring in the 'aromatic carbocycle', 'non-aromatic carbocycle', and/or 'aromatic heterocycle' is condensed.

The term 'halogen' as used herein may be fluorine, chlorine, bromine, or iodine.

The term 'hetero' as used herein, unless otherwise stated, refers to the same or different heteroatoms randomly selected from oxygen atoms, sulfur atoms and nitrogen atoms.

The term 'substituted' as used herein, unless otherwise stated, refers to being substituted with one or more substituents selected from halogen, hydroxy, amino, cyano, nitro, alkyl, alkenyl, haloalkyl, (mono-, di-, or trihalogen)alkyl, hydroxyalkyl, alkoxy, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, carboxamide(-CONH ₂ ), alkylcarbonyl, alkylamino, dialkylamino, alkylcarbonylamino, alkylcarbonylaminoalkyl, alkyloxycarbonyl, alkyloxycarbonylalkyl, alkyloxycarbonyloxy, alkylsulfanyl, and alkylsulfonyl.

The term 'pharmaceutically acceptable salt' in this specification, unless otherwise stated, refers to any salt that has low toxicity to the human body and does not adversely affect the biological activity and physicochemical properties of the parent compound, and includes acid or base addition salts.

As used herein, the term 'optical isomers' includes a pair of stereoisomers that are non-superimposable mirror images of the compounds of the present invention, unless otherwise stated.

As used herein, the term 'solvate', unless otherwise stated, refers to a molecular complex comprising one or more pharmaceutically acceptable solvent molecules.

As used herein, the term 'hydrate' refers to a molecular complex in which the solvent molecule is water, unless otherwise stated.

The numerical range indicated using the term 'to' in this specification refers to a range that includes the numerical values described before and after as the lower and upper limits, respectively, unless otherwise stated.

The terms “have,” “may have,” “include,” or “may include” used in this specification indicate the presence of a feature (e.g., a numerical value, or a component such as an ingredient), and do not exclude the presence of additional features.

The present invention is described in more detail below.

According to one aspect of the present invention, the present invention provides any one compound selected from the following chemical formula I compound and its pharmaceutically acceptable salts, optical isomers, hydrates and solvates:

[Chemical Formula I]

In the above chemical formula I,

Ring A is any one nitrogen-containing heterocycle selected from the group consisting of A-1 to A-5 below;

In the above A-1 to A-5,

Wavy lines ( ) indicates the attachment site;

A plurality of X ⁰ are different from each other, one of which is C and the rest are N;

A plurality of X ¹ are identical or different, and each independently is C(R) or N, or at least one is N;

A plurality of X ² are different from each other, one of which is C(R') and the rest are N;

X ³ is C(R'') or N;

R and R'' are each independently hydrogen, halogen, C ₁ -C ₃ alkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;

R' is halogen, C ₁ -C ₃ alkyl, carboxamide (-CONH ₂ ), substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;

R ³ is hydrogen, or a substituted or unsubstituted amino group;

The substituent of R ³ is a substituted or unsubstituted C ₁ -C ₆ alkyl, a substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, a substituted or unsubstituted 3 to 10 membered heterocycloalkyl, a substituted or unsubstituted C ₆ -C ₁₂ aryl, or a substituted or unsubstituted 5 to 12 membered heteroaryl;

R ⁴ to R ⁷ are each independently hydrogen, halogen, C ₁ -C ₃ alkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;

When each of R ⁴ to R ⁷ is alkyl, aryl or heteroaryl, their substituents may each independently be hydroxy, hydroxy C ₁ -C ₆ alkyl, or C ₁ -C ₆ alkoxy;

R ⁸ is hydrogen or halogen;

Ring B is aryl or heteroaryl substituted or unsubstituted with R ¹ ;

R ¹ is each independently hydrogen, hydroxy, cyano, nitro, (mono-, di-, or trihalogen)methyl, halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkene, C ₂ -C ₆ alkyne, C ₁ -C ₃ alkoxy, C ₃ -C ₆ cycloalkyl, or C ₁ -C ₆ dialkylamino;

L ¹ is C _0- C ₆ alkylene, C _2- C ₆ alalkenylene, C _2- C ₆ alkynylene, -NR ⁹ -CO-, -CO-NR ¹⁰ -, -NR ⁹ -CO-NR ¹⁰ -, -NR ⁹ -SO ₂ -, -NR ⁹ -SO ₂ -NR ¹⁰ -, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene, cycloalkylene or heterocycloalkylene, or may be a direct bond;

L ² is -NR ⁹ -CO-, -CO-NR ¹⁰ -, -NR ⁹ -SO ₂ -, or -NR ⁹ -SO-NR ¹⁰ -;

R ⁹ and R ¹⁰ are each independently hydrogen or C ₁ -C ₆ alkyl;

Ring C is aryl or heteroaryl substituted or unsubstituted with R ² ;

R ² can be independently selected from the group consisting of hydrogen, hydroxy, hydroxy C ₁ -C ₆ alkyl, cyano, C _1- C ₆ alkyl, C _2- C ₆ alkene, C _2- C ₆ alkyne, C _1- C ₃ alkoxy, (mono-, di-, or trihalogen)methyl, halogen, C _3- C ₆ cycloalkyl, C _1- C ₆ dialkylamino, substituted or unsubstituted aryloxy or heteroaryloxy, substituted or unsubstituted arylalkyloxy or heteroarylalkyloxy, substituted or unsubstituted aryl or heteroaryl, and substituted or unsubstituted cycloalkyl or heterocycloalkyl;

p and q are 0 and 1 respectively, and p+q=1;

m is an integer from 0 to 4;

n is an integer from 0 to 5;

When R ¹ or R ² is plural, they are each the same or different.

In one embodiment, A-1 may be any one heteroaryl selected from the group below.

In one embodiment, the ring B may be an arylene substituted or unsubstituted with a halogen.

In one embodiment, the ring C can be phenyl or pyridine, which is optionally substituted with R ² .

In one embodiment, L ¹ can be a direct bond, C _2- C ₆ alkynylene or -NH-CO-.

In one implementation example, L ² can be -NH-CO- or -NH-SO ₂ -.

In one embodiment, L ¹ may be C _2- C ₆ alkynylene or -NH-CO-, and L ² may be -NH-CO- or -NH-SO ₂ -.

In one implementation example,

R' is halogen, C ₁ -C ₃ alkyl, carboxamide (-CONH ₂ ), C ₆ -C ₁₂ aryl, or 5 to 12 membered heteroaryl, or

each independently selected halogen, hydroxy, C ₁ -C ₃ alkyl, or C ₁ -C ₃ alkoxy substituted

It may be C ₆ -C ₁₂ aryl or 5 to 12 membered heteroaryl.

In one implementation example,

R' is halogen, C ₁ -C ₃ alkyl, carboxamide (-CONH ₂ ), phenyl, pyridine, or thiazole, or

each independently selected halogen, hydroxy, C ₁ -C ₃ alkyl, or C ₁ -C ₃ alkoxy substituted

It can be phenyl, pyridine, or thiazole.

In one implementation example,

R ³ is hydrogen or an amino group, or

unsubstituted or substituted with halogen, hydroxy, C ₁ -C ₆ alkyl, or C ₁ -C ₆ alkoxy, each independently selected

It may be an amino group substituted with C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl, 3 to 10 membered heterocycloalkyl, C ₆ -C ₁₂ aryl, or 5 to 12 membered heteroaryl.

In one implementation example,

R ⁵ to R ⁶ can each independently be hydrogen, halogen, C ₁ -C ₃ alkyl, hydroxy C ₁ -C ₃ alkyl, or (C ₁ -C ₃ alkoxy) C ₁ -C ₃ alkyl.

The present invention may be a compound selected from the group consisting of compounds in Table 1 below.

[Table 1]

The present invention also provides pharmaceutically acceptable salts, optical isomers, hydrates, or solvates of the compound represented by the above formula (I). Pharmaceutically acceptable salts should have low toxicity to the human body and should not adversely affect the biological activity or physicochemical properties of the parent compound. Pharmaceutically acceptable salts include, but are not limited to, acid addition salts of pharmaceutically acceptable free acids and base compounds of formula (I).

Pharmaceutically acceptable salts of the compound according to the present invention include, for example, salts of the compound according to the present invention with alkali metals (e.g., lithium, sodium, potassium, etc.), alkaline earth metals (e.g., calcium, barium, etc.), magnesium, transition metals (e.g., zinc, iron, etc.), ammonia, organic bases (e.g., trimethylamine, triethylamine, dicyclohexylamine, ethanolamine, diethanolamine, triethanolamine, meglumine, ethylenediamine, pyridine, picoline, quinoline, etc.) and amino acids, or salts of inorganic acids (e.g., hydrochloric acid, sulfuric acid, nitric acid, carbonic acid, hydrobromic acid, phosphoric acid, hydroiodic acid, etc.), and organic acids (e.g., formic acid, acetic acid, propionic acid, trifluoroacetic acid, citric acid, lactic acid, tartaric acid, oxalic acid, maleic acid, fumaric acid, mandelic acid, glutaric acid, malic acid, benzoic acid, phthalic acid, Examples include salts with ascorbic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, etc. In particular, salts with hydrochloric acid, sulfuric acid, phosphoric acid, tartaric acid, and methanesulfonic acid are examples. These salts can be formed by conventional methods.

The present invention provides a pharmaceutical composition comprising a compound represented by the above chemical formula I, a pharmaceutically acceptable salt, optical isomer, hydrate or solvate thereof as an active ingredient, and in particular, provides a pharmaceutical composition having an action of inhibiting abnormal activity of eIF2α (more specifically, GCN2), and provides a pharmaceutical composition for preventing or treating cancer, immune disease or Alzheimer's disease caused by abnormal activity of GCN2.

The present invention also provides a method for treating or preventing a disease caused by abnormal activity of GCN2, comprising administering a therapeutically effective amount of a compound represented by the above chemical formula I, or a pharmaceutically acceptable salt, optical isomer, hydrate or solvate thereof, to a person in need of treatment.

Hereinafter, a manufacturing method for an exemplary compound of chemical formula I according to the present invention will be described.

The compound of the above formula I according to the present invention can be prepared using chemical transformations well known to those skilled in the art of organic/medicinal chemistry, according to methods representatively illustrated in the following reaction schemes 1 and 2.

[Reaction Formula 1]

The compounds of the above chemical formulas II and II' and the compounds of the above chemical formulas III and III' can be prepared using common knowledge in the field of organic chemistry.

In the above reaction formula 1,

A, B, C, R ¹ , R ² , L ² , m and n are as defined in the above chemical formula I.

In order to prepare a compound of formula I by reacting the compounds of formulae II and II' with the compounds of formulae III and III', the solvent used in the reaction may be any solvent that does not inhibit the reaction. The solvent used in the reaction may be a polar aprotic solvent such as dimethyl sulfoxide, N , N -dimethyl formamide, acetonitrile, or tetrahydrofuran; a polar protic solvent such as methanol, ethanol, 2-propanol, 2-isopropanol, or 2-butanol; or a nonpolar aprotic solvent such as toluene or 1,4-dioxane. Specifically, compound I was prepared by reacting a halogen-substituted compound II and a palladium catalyst such as (trimethylphosphine)palladium (II) chloride, a ligand such as cuprous iodide, and an organic base such as triethylamine in an organic solvent such as N , N -dimethylformamide at a temperature range of about 80°C. However, when A contained a protecting group, compound I was prepared by deprotection using trifluoroacetic acid. The reaction temperature may be from 0°C to room temperature, and preferably 0°C.

[Reaction Formula 2]

The compound of the above chemical formula IV and the compound of the above chemical formula V can be prepared using common knowledge in the field of organic chemistry.

In the above reaction formula 2,

A, B, C, R ¹ , R ² , L ¹ , L ² , m and n are as defined in the above chemical formula I.

In order to prepare a compound of formula VI by reacting the compound of formula IV with the compound of formula V, the solvent used in the reaction may be any solvent that does not inhibit the reaction, and specifically, the reaction may be performed by adding or not adding an organic base such as triethylamine, diisopropylethylamine, or pyridine to the reaction solution; or an inorganic base such as sodium carbonate, potassium carbonate, or sodium hydride, and at this time, the preferred amount of the base is 1.5 to 2 equivalents based on 1 equivalent of the compound of formula IV. The solvent used in the reaction may be a polar aprotic solvent such as dimethyl sulfoxide, N , N -dimethyl formamide, acetonitrile, or tetrahydrofuran; a polar protic solvent such as methanol, ethanol, 2-propanol, or 2-butanol; or a nonpolar aprotic solvent such as toluene or 1,4-dioxane. The reaction temperature may be 0°C to room temperature, and preferably room temperature.

The compound of the above chemical formula VI was reduced using a reducing agent such as iron in an organic solvent such as ethanol to prepare a compound of the chemical formula VII. The reaction temperature may be 0°C to 150°C, and preferably 80°C to 90°C.

The compound of formula I was prepared by condensation reaction of the compound of formula VII with the compound of formula VIII in an organic base such as pyridine. The reaction temperature may be 0°C to room temperature, and preferably room temperature.

Hereinafter, the present invention will be described in more detail with the following examples and experimental examples. However, these examples and experimental examples are intended only to aid understanding of the present invention and are not intended to limit the scope of the present invention in any way.

[Example]

Example 1: N -(3-((8-Aminoimidazo[1,2- a Preparation of ]pyrazin-5-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide

Step 1) 5-bromoimidazo[1,2- a ] Preparation of pyrazin-8-amine

5,8-Dibromoimidazo[1,2- a ]pyrazine (500 mg, 1.81 mmol) was diluted in 7.5 mL of ammonia water and stirred at 90°C for 14 hours in a sealed tube. Upon completion of the reaction, the resulting reaction mixture was cooled to room temperature, and the resulting solid was washed with distilled water, filtered under reduced pressure, and dried under reduced pressure to obtain 250 mg (yield: 65%) of the title compound.

¹ H-NMR (300 MHz, DMSO- d ₆ ): δ 7.90 (s, 1H), 7.62 (d, 1H), 7.39 (s, 1H), 7.09 (brs, 2H).

Step 2) 5-((trimethylsilyl)ethynyl)imidazo[1,2- a ] Preparation of pyrazin-8-amine

500 mg (2.35 mmol) of the compound prepared in the above step 1) and 41 mg (0.06 mmol) of bis(triphenylphosphine)palladium(II) dichloride, 16 mg (0.08 mmol) of cuprous iodide, 514 μL (2.58 mmol) of dicyclohexylamine, and 663 μL (4.69 mmol) of trimethylsilylacetylene were diluted in 10 mL of acetonitrile and stirred at 80°C for 16 hours in a sealed tube. Upon completion of the reaction, the resulting reaction mixture was cooled to room temperature, washed with ethyl acetate through a Celite-packed filter, filtered under reduced pressure, and the resulting filtrate was washed with distilled water and saturated brine. The separated organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure. The resulting residue was separated by column chromatography (dichloromethane: methanol = 40:1 (volume ratio)) to obtain 210 mg (yield: 39%) of the title compound.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 7.73 (d, 1H), 7.60 (d, 2H), 5.69 (br s, 2H), 0.31 (s, 9H).

Step 3) 5-Ethynylimidazo[1,2- a ] Preparation of pyrazin-8-amine

210 mg (0.91 mmol) of the compound prepared in the above step 2) was diluted in 4 mL of tetrahydrofuran, 1.82 mL (1.82 mmol) of tetra- n -butylammonium fluoride (1 M tetrahydrofuran solution) was added, and the mixture was stirred at room temperature for 1 hour. Upon completion of the reaction, the resulting reaction mixture was diluted with ethyl acetate and washed with distilled water and a saturated aqueous sodium bicarbonate solution. The resulting organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure to obtain 100 mg (yield: 69%) of the title compound.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 7.77 (d, 1H), 7.61 (d, 2H), 5.69 (br s, 2H), 3.70 (s, 3H).

Step 4) Preparation of 3-(benzylthio)-5-chloro-2-methoxypyridine

6.72 g (30.2 mmol) of 3-bromo-5-chloro-2-methoxypyridine, 3.54 mL (30.2 mmol) of benzyl mercaptan, 692 mg (0.76 mmol) of tris(dibenzylideneacetone)dipalladium(O), 874 mg (1.51 mmol) of 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene, and 10.6 mL (60.4 mmol) of N,N -diisopropylethylamine were diluted in 300 mL of 1,4-dioxane and stirred at 100 °C for 1 hour. Upon completion of the reaction, the resulting reaction mixture was cooled to room temperature, distilled under reduced pressure, diluted with ethyl acetate, and washed with a saturated aqueous sodium bicarbonate solution and saturated brine. The resulting organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure. The resulting residue was separated by column chromatography (hexane: ethyl acetate = 40:1 (volume ratio)) to obtain 8.2 g (yield: 100%) of the title compound.

Step 5) Preparation of 5-chloro-2-methoxypyridine-3-sulfonyl chloride

8.02 g (30.2 mmol) of the compound prepared in the above step 4) was diluted in 150 mL of acetonitrile, and 17.4 mL (302.0 mmol) of acetic acid and 7.5 mL (416.7 mmol) of distilled water were added at room temperature. Then, 11.9 g (60.6 mmol) of 1,3-dichloro-5,5-dimethyl hydantoin was slowly added at 0 °C and stirred for 30 minutes. After the reaction was completed, the resulting reaction mixture was distilled under reduced pressure, diluted in ethyl acetate, and washed with distilled water and a saturated sodium bicarbonate aqueous solution. The separated organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure. The resulting residue was separated by column chromatography (hexane: ethyl acetate = 20: 1 (volume ratio)) to obtain 5.3 g (yield: 73%) of the title compound.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.42 (d, 1H), 8.21 (d, 1H), 4.18 (s, 3H).

Step 6) Preparation of 1,3-difluoro-2-iodo-4-nitrobenzene

9.0 g (37.5 mmol) of 2,6-difluoroiodobenzene was diluted in 30 mL of sulfuric acid, and 6.5 mL (93.8 mmol) of nitric acid was slowly added at 0 °C, followed by stirring at room temperature for 30 minutes. After the reaction was completed, the resulting reaction mixture was diluted with cold distilled water, neutralized with 5% aqueous sodium hydroxide solution, diluted with ethyl acetate, and washed with saturated brine. The separated organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure to obtain 10.4 g (yield: 97%) of the title compound.

Step 7) Preparation of 2,4-difluoro-3-iodoaniline

10.4 g (36.5 mmol) of the compound prepared in the above step 6) and 24.7 g (109.5 mmol) of stannous chloride were diluted in 40 mL of hydrochloric acid and stirred at 50°C for 1 hour. After the reaction was completed, the resulting reaction mixture was cooled to room temperature, diluted with distilled water, neutralized with a 5% aqueous sodium hydroxide solution, diluted with dichloromethane, and washed with saturated brine. The resulting organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure. The resulting residue was separated by column chromatography (hexane: ethyl acetate = 10: 1 (volume ratio)) to obtain 6.0 g (yield: 65%) of the title compound.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 6.77 (m, 2H), 3.68 (br s, 2H).

Step 8) 5-Chloro- N Preparation of -(2,4-difluoro-3-iodophenyl)-2-methoxypyridine-3-sulfonamide

930 mg (3.84 mmol) of the compound prepared in the above step 5) and 980 mg (3.84 mmol) of the compound prepared in the above step 7) were diluted in 10 mL of pyridine and stirred at room temperature for 2 hours. When the reaction was completed, the resulting reaction mixture was diluted in 2 mL of methanol and ethyl acetate and washed with distilled water. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered under reduced pressure, and distilled under reduced pressure. The resulting residue was separated by column chromatography (hexane: ethyl acetate = 10: 1 (volume ratio)) to obtain 1.68 g (yield: 95%) of the title compound.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.26 (d, 1H), 8.02 (d, 1H), 7.59 (m, 1H), 6.89 (m, 1H), 4.12 (s, 3H).

Step 9) N -(3-((8-Aminoimidazo[1,2- a Preparation of ]pyrazin-5-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide

126 mg (0.35 mmol) of the compound prepared in the above step 8) and 50 mg (0.316 mmol) of the compound prepared in the above step 3), 12 mg (0.02 mmol) of bis(triphenylphosphine)palladium(II) dichloride, 7 mg (0.04 mmol) of cuprous iodide, and 245 μL (1.76 mmol) of triethylamine were diluted in 3 mL of N , N -dimethylformamide and stirred at 80°C for 2 hours. Upon completion of the reaction, the resulting reaction mixture was cooled to room temperature, washed with ethyl acetate through a Celite-packed filter, filtered under reduced pressure, and then the resulting filtrate was washed with distilled water and saturated brine. The separated organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure. The resulting residue was separated by column chromatography (dichloromethane: methanol = 40:1 (volume ratio)) to obtain 40 mg of the title compound (final stage yield: 23%).

¹ H-NMR (300MHz, DMSO- d ₆ ): δ 10.5 (br s, 1H), 8.51 (d, 1H), 8.08 (d, 1H), 7.77 (m, 1H), 7.70 (m, 1H), 7.67 (m, 1H), 7.64 (br s, 2H), 7.38 (q, 1H), 7.35 (t, 1H), 3.93 (s, 3H);

MS (ESI ⁺ ): m/z = 491.14 [M+H] ⁺ .

The compounds of Examples 2 to 13 shown in Table 2 below were each prepared using the same or similar method as that of Example 1.

[Table 2]

Example 14: N Preparation of -(3-((2-amino-4-(pyridin-3-yl)pyrimidin-5-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide

Step 1) Preparation of 4-(pyridin-3-yl)pyrimidin-2-amine

1.0 g (7.7 mmol) of 2-amino-4-chloropyrimidine and 1.0 g (8.5 mmol) was diluted with 40 mL of a mixture of acetonitrile: water (1:1 (volume ratio)), 898 mg (8.5 mmol) of sodium carbonate and 162 mg (0.23 mmol) of bis(triphenylphosphine)palladium(II) dichloride were added, and the mixture was stirred at 80°C for 12 hours. After the reaction was completed, the resulting reaction mixture was cooled to room temperature, washed with ethyl acetate through a Celite-packed filter, and filtered under reduced pressure. The resulting filtrate was washed sequentially with a saturated aqueous ammonium chloride solution and saturated brine. The resulting organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure. The obtained solid was diluted with 10 mL of dichloromethane, washed with dichloromethane, and filtered under reduced pressure. The resulting solid was dried under reduced pressure to obtain 800 mg (yield: 60%) of the title compound.

^1H -NMR (300MHz, DMSO- d ₆ ): δ 9.21 (s, 1H), 8.66 (m, 1H), 8.38 (m, 1H), 8.34 (m, 1H), 7.52 (m, 1H), 7.19 (m, 1H), 6.74 (s, 2H).

Step 2) Preparation of 5-iodo-4-(pyridin-3-yl)pyrimidin-2-amine

800 mg (4.65 mmol) of the compound prepared in the above step 1) was diluted in 16 mL of N,N -dimethylformamide, 1.6 g (6.97 mmol) of N -iodosuccinimide was added, and the mixture was stirred at 80°C for 12 hours. Upon completion of the reaction, the resulting reaction mixture was cooled to room temperature, diluted with ethyl acetate, and washed with an aqueous sodium bicarbonate solution. The resulting organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure. The resulting residue was separated by column chromatography (dichloromethane: methanol = 10:1 (volume ratio)) to obtain 417 mg (yield: 61%) of the title compound.

^1H -NMR (300MHz, DMSO- d ₆ ): δ 8.31 (m, 1H), 8.29 (m, 1H), 7.21 (m, 1H), 7.15 (m, 1H), 6.93 (m, 1H), 6.79 (m, 1H), 6.60 (m, 1H), 3.71 (s, 2H).

Step 3) Preparation of 2,4-difluoro-3-((trimethylsilyl)ethynyl)aniline

The same process as in step 2) of Example 1 was performed except that the compound prepared in step 7) of Example 1 was used instead of 5-bromoimidazo[1,2- a ]pyrazin-8-amine in step 2) of Example 1, to obtain 7.6 g (yield: 87%) of the title compound.

Step 4) Preparation of 3-ethynyl-2,4-difluoroaniline

The same process as in step 3) of Example 1 was performed except that the compound prepared in step 3) was used instead of 5-((trimethylsilyl)ethynyl)imidazo[1,2- a ]pyrazin-8-amine in step 3) of Example 1, to obtain 3.4 g (yield: 66%) of the title compound.

¹ H-NMR (300 MHz, DMSO- d ₆ ): δ 6.74 (m, 2H), 3.63 (bs, 2H), 3.49 (s, 1H).

Step 5) 5-chloro- N Preparation of -(3-ethynyl-2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide

The same process as in step 8) of the above Example 1 was performed except that the compound prepared in step 4) was used instead of 2,4-difluoro-3-iodoaniline in step 8) of the above Example 1, to obtain 6.0 g (yield: 90%) of the title compound.

Step 6) N Preparation of -(3-((2-amino-4-(pyridin-3-yl)pyrimidin-5-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide

The same process as in step 9) of Example 1 was performed except that the compound prepared in step 2) was used instead of 5-chloro- N- (2,4-difluoro-3-iodophenyl)-2-methoxypyridine-3-sulfonamide in step 9) of Example 1 and the compound prepared in step 5) was used instead of 5-ethynylimidazo[1,2- a ]pyrazin-8-amine, to obtain 29 mg (yield: 22%) of the title compound.

¹ H-NMR (300 MHz, DMSO- d ₆ ): δ 10.42 (s, 1H), 9.09 (s, 1H), 8.70 (s, 1H), 8.56 (s, 1H), 8.51 (m, 1H), 8.31 (d, 1H), 8.05 (m, 1H), 7.47 (m, 3H), 7.31 (m, 1H), 7.12 (m, 1H), 3.92 (s, 3H).

MS (ESI ⁺ ): m/z = 529.06 [M+H] ⁺ .

The compounds of Examples 15 to 24 shown in Table 3 below were each prepared using a method identical or similar to that of Example 14.

[Table 3]

Example 25: N Preparation of -(2-amino-4-(pyridin-3-yl)pyrimidin-5-yl)-3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6-difluorobenzamide

Step 1) Preparation of 5-((diphenylmethylene)amino)-4-(pyridin-3-yl)pyrimidin-2-amine

1.33 g (4.48 mmol) of the compound prepared in the above Example 14 Step 2) and 0.812 g (4.48 mmol) of benzophenone imine were dissolved in 30 mL of 1,4-dioxane, and then 0.205 g (0.224 mmol) of tris(dibenzylideneacetone)dipalladium(0), 0.254 g (0.448 mmol) of 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene, and 4.37 g (13.44 mmol) of cesium carbonate were added, and the mixture was stirred at 105 °C for 18 hours. Upon completion of the reaction, the resulting reaction mixture was cooled to room temperature and filtered through a filter filled with Celite. The filtrate was diluted with ethyl acetate and washed with water. The separated organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure. The resulting residue was separated by column chromatography (dichloromethane: methanol = 10:1 (volume ratio)) to obtain 1.12 g (yield: 72%) of the title compound.

¹ H-NMR (300 MHz, DMSO- d ₆ ): δ 8.95 (s, 1H), 8.56 (s, 1H), 8.15 (m, 1H), 7.64 (m, 3H), 7.52 (m, 1H), 7.46 (m, 3H), 7.36 (m, 3H), 7.01(m, 2H), 6.51 (s, 2H).

Step 2) Preparation of 4-(pyridin-3-yl)pyrimidine-2,5-diamine

1.12 g (3.21 mmol) of the compound prepared in the above step 1) was diluted in 30 mL of methanol, 0.446 g (6.42 mmol) of hydroxylamine hydrochloride and 0.787 g (8.02 mmol) of potassium acetate were added, and the mixture was stirred at 45°C for 16 hours. After the reaction was completed, the resulting reaction mixture was diluted with a mixed solvent of chloroform: 2-propanol (3: 1 (volume ratio)) and washed with saturated sodium bicarbonate. The separated organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure. The obtained residue was separated by column chromatography (chloroform: methanol = 8: 1 (volume ratio)) to obtain 250 mg (yield: 41%) of the title compound.

¹ H-NMR (300 MHz, DMSO- d ₆ ): δ 9.05 (s, 1H), 8.70 (m, 1H), 8.13 (m, 1H), 7.44 (m, 1H), 4.74 (s, 2H), 3.37 (s, 2H).

Step 3) Preparation of methyl-3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6-difluorobenzoate

The same process as in step 8) of the above Example 1 was performed except that methyl 3-amino-2,6-difluorobenzoate was used instead of 2,4-difluoro-3-iodoaniline in step 8) of the above Example 1, to obtain 610 mg (yield: 75%) of the title compound.

Step 4) Preparation of 3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6-difluorobenzoic acid

603 mg (1.5 mmol) of the compound prepared in the above step 3) was diluted with 6 mL of a mixed solvent of tetrahydrofuran: methanol: distilled water (3:3:1 (volume ratio)), 194 mg (4.6 mmol) of lithium hydroxide monohydrate was added, and the mixture was stirred at room temperature for 12 hours. When the reaction was complete, the resulting reaction mixture was cooled to 0°C, diluted with a mixed solvent of chloroform: 2-propanol (3:1 (volume ratio)), and acidified with 1 N hydrochloric acid aqueous solution (pH 2). The resulting separated organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure to obtain 500 mg (yield: 86%) of the title compound.

Step 5) N Preparation of -(2-amino-4-(pyridin-3-yl)pyrimidin-5-yl)-3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6-difluorobenzamide

74 mg (0.2 mmol) of the compound prepared in step 4) above was diluted in 1 mL of dichloromethane, 0.037 mL (0.3 mmol) of oxalyl chloride and 10 μL of N , N -dimethylformamide were added, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the resulting reaction mixture was distilled under reduced pressure and dried under reduced pressure to prepare 3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6-difluorobenzoyl chloride. 19 mg (0.1 mmol) of the compound prepared in step 2) of Example 12 above was diluted in 1 mL of tetrahydrofuran, cooled to 0°C, 0.024 g (0.2 mmol) of sodium hydride was slowly added, and the mixture was stirred for 15 minutes. The reaction mixture was added with the above-mentioned 3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6-difluorobenzoyl chloride and stirred at room temperature for 16 hours. After the reaction was completed, the resulting reaction mixture was diluted with ethyl acetate and washed with saturated brine. The resulting organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure. The resulting residue was separated by column chromatography (ethyl acetate: hexane = 1:40 (volume ratio)) to obtain 0.004 g (yield: 7%) of the title compound.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.89 (s, 1H), 8.63 (s, 1H), 8.36 (m, 1H), 8.30 (m, 1H), 8.15 (m, 1H), 8.07 (m, 1H), 7.59 (m, 2H), 7.05 (t, 1H), 4.02 (s, 3H).

MS (ESI ⁺ ): m/z = 548.06 [M+H] ⁺ .

Example 26: N -(3-((4-amino-1 H -Pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide manufacturing

Step 1) N -(4-methoxybenzyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1 H -Pyrazolo[4,3- c ] Preparation of pyridin-4-amine

1.2 g (4.16 mmol) of 4-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1 H -pyrazolo[4,3- c ]pyridine (WO2018011628) was diluted in 7.2 mL of 4-methoxybenzylamine and stirred at 130 ℃ for 2 hours in a sealed tube. Upon completion of the reaction, the resulting reaction mixture was cooled to room temperature, diluted in ethyl acetate, and washed with distilled water and a saturated ammonium chloride aqueous solution. The separated organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure. The resulting residue was purified by column chromatography (hexane: ethyl acetate = 10: 1 (volume ratio)) to obtain 1.1 g (yield: 69%) of the title compound.

Step 2) 7-Iodo- N -(4-methoxybenzyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1 H -Pyrazolo[4,3- c ] Preparation of pyridin-4-amine

1.1 g (2.86 mmol) of the compound prepared in the above step 1) was diluted in 27.5 mL of N,N -dimethylformamide, 965 mg (4.29 mmol) of N -iodosuccinimide was added, and the mixture was stirred at room temperature for 1 hour. When the reaction was complete, the resulting reaction mixture was diluted with ethyl acetate and washed with a saturated aqueous sodium bicarbonate solution. The resulting organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure. The residue obtained was separated by column chromatography (hexane: ethyl acetate = 10: 1 (volume ratio)) to obtain 980 mg (yield: 67%) of the title compound.

Step 3) N -(3-((4-amino-1-((2-(trimethylsilyl)ethoxy)methyl)-1 H -Pyrazolo[4,3- c Preparation of ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide

The same procedure as step 3) of Example 12 was sequentially performed, except that the compound prepared in step 2) was used instead of 5-iodo-4-(pyridin-3-yl)pyrimidin-2-amine in step 6) of Example 14, to obtain 40 mg (yield: 55%) of the title compound.

Step 4) N -(3-((4-amino-1 H -Pyrazolo[4,3- c Preparation of ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide

40 mg (0.054 mmol) of the compound prepared in step 3) above was dissolved in 1 mL of dichloromethane, 1 mL of trifluoroacid was added, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the resulting reaction mixture was distilled under reduced pressure, and the obtained residue was diluted in 1 mL of tetrahydrofuran, 1 mL of 6N aqueous sodium hydroxide solution was added, and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, the resulting reaction mixture was diluted with dichloromethane and washed with distilled water and a saturated aqueous ammonium chloride solution. The separated organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure. The obtained residue was separated by column chromatography (dichloromethane: methanol = 8: 1 (volume ratio)) to obtain 10 mg (final yield: 38%) of the title compound.

¹ H-NMR (300MHz, DMSO- d ₆ ): δ 13.4 (br s, 1H), 8.49 (d, 1H), 8.24 (br s, 1H), 8.06 (d, 1H), 7.91 (s, 1H), 7.34 (m, 4H), 7.20 (m, 1H), 3.94 (s, 3H).

MS (ESI ⁺ ): m/z = 491.04 [M+H] ⁺ .

The compounds of Examples 27 to 44 shown in Table 4 below were each prepared using the same or similar method as that of Example 26.

[Table 4]

Example 45: N -(4-amino-1 H -Pyrazolo[4,3- c ]pyridin-7-yl)-3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6-difluorobenzamide manufacturing

Step 1) 2,6-Difluoro- N -(4-((4-methoxybenzyl)amino)-1-((2-(trimethylsilyl)ethoxy)methyl)-1 H -Pyrazolo[4,3- c Preparation of ]pyridin-7-yl)-3-nitrobenzamide

Except that the compound prepared in step 2) of Example 26 was used instead of 5-iodo-4-(pyridin-3-yl)pyrimidin-2-amine in step 1) of Example 25 and that 2,6-difluoro-3-nitrobenzoic acid was used instead of 3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6-difluorobenzoic acid in step 5) of Example 25, the same processes as steps 1), 2), and 5) of Example 25 were performed sequentially to obtain 929 mg of the title compound (final yield: 64%).

¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.94 (s, 1H), 8.64 (s, 1H), 8.27 (m, 1H), 7.87 (s, 1H), 7.39 (m, 2H), 7.20 (m, 1H), 6.93 (m, 2H), 5.76 (s, 2H), 5.24 (m, 1H), 4.78 (d, 2H), 3.83 (s, 3H), 3.53 (t, 2H), 0.78 (t, 2H), -0.09 (s,9H).

Step 2) 3-Amino-2,6-difluoro- N -(4-((4-methoxybenzyl)amino)-1-((2-(trimethylsilyl)ethoxy)methyl)-1 H -Pyrazolo[4,3- c Preparation of ]pyridin-7-yl)benzamide

929 mg (1.59 mmol) of the compound prepared in the above step 1) was diluted with 27 mL of a mixed solvent of tetrahydrofuran: methanol: distilled water (1: 1: 1 (volume ratio)), 532 mg (9.53 mmol) of iron and 929 mg (1.59 mmol) of ammonium chloride were added, and the mixture was stirred at 90°C for 2 hours. After the reaction was completed, the resulting reaction mixture was filtered through a Celite-packed filter to remove iron, and the resulting filtrate was distilled under reduced pressure. The resulting residue was diluted with dichloromethane and washed with a saturated sodium bicarbonate aqueous solution. The separated organic layer was dried over anhydrous sodium sulfate, and the resulting residue was purified by column chromatography (ethyl acetate: hexane = 1: 1 (volume ratio)) to obtain 822 mg (yield: 94%) of the title compound.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 8.64 (s, 1H), 8.60 (s, 1H), 7.86 (s, 1H), 7.39 (m, 2H), 6.92 (m, 2H), 6.84 (m, 2H), 5.78 (s, 2H), 5.10 (m, 1H), 4.78 (d, 2H), 3.84 (s, 3H), 3.74 (s, 2H), 3.50 (t, 2H), 0.91 (t, 2H), -0.08 (s, 9H).

Step 3) N -(4-amino-1 H -Pyrazolo[4,3- c ]pyridin-7-yl)-3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6-difluorobenzamide manufacturing

Except for using the compound prepared in step 2) instead of 3-ethynyl-2,4-difluoroaniline in step 5) of the above Example 14, the same process as step 5) of the above Example 14 and step 4) of the above Example 26 was performed sequentially to obtain 929 mg of the title compound (final yield: 64%).

1H-NMR (300MHz, DMSO- d ₆ ): δ 10.54 (s, 1H), 8.52 (m, 1H), 8.38 (s, 1H), 8.10 (m, 1H), 7.89 (s, 1H), 7.43 (m, 1H), 7.22 (m, 1H), 3.97 (s, 3H).

MS (ESI ⁺ ): m/z = 510.05 [M+H] ⁺ .

Example 46: N -(4-amino-1 H -Pyrazolo[4,3- c Preparation of ]pyridin-7-yl)-2,6-difluoro-3-((2-methoxy-5-(trifluoromethyl)pyridine)-3-sulfonamido)benzamide

The same process as in Example 45 was performed except that 2-methoxy-5-(trifluoromethyl)pyridine-3-sulfonyl chloride was used instead of 5-chloro-2-methoxypyridine-3-sulfonyl chloride in step 3) of Example 45, to obtain 929 mg of the title compound (final yield: 64%).

^1H -NMR (300MHz, DMSO- d ₆ ): δ 10.55 (s, 1H), 8.90 (s, 1H), 8.42 (m, 1H), 8.29 (s, 1H), 7.90 (m, 1H), 7.46 (m, 1H), 7.22 (m, 1H), 4.04 (s,3H).

MS (ESI ⁺ ): m/z = 525.07 [M+H] ⁺ .

Example 47: Preparation of 3-amino-6-((3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6-difluorophenyl)ethynyl)pyrazine-2-carboxamide manufacturing

Except for using methyl 3-aminopyrazine-2-carboxylate instead of 4-(pyridin-3-yl)pyrimidin-2-amine in step 2) of Example 14, the same method as step 2) of Example 14, step 1) of Example 1, and step 6) of Example 14 was performed sequentially to obtain 75 mg of the title compound (final yield: 27%).

¹ H-NMR (300 MHz, DMSO- d ₆ ): δ 10.8 (br s, 1H), 8.49 (d, 1H), 8.40 (s, 1H), 8.05 (s, 2H), 7.69 (s, 1H), 7.33 (m, 1H), 7.23 (t, 1H), 3.92 (s, 3H).

MS (ESI ⁺ ): m/z = 495.04 [M+H] ⁺ .

Example 48: 3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6-difluoro- N -(5 H -Pyrrolo[2,3- b Preparation of ]pyrazin-2-yl)benzamide

Step 1) 3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6-difluoro- N -(5-Tosil-5 H -Pyrrolo[2,3- b Preparation of ]pyrazin-2-yl)benzamide

Except that 5-tosyl-5 H -pyrrolo [2,3- b ]pyrazin-2-amine (WO2013170072) was used instead of N ⁴ -(4-methoxybenzyl)-1-((2-(trimethylsilyl)ethoxy)methyl) -1 H -pyrazolo [4,3- c ]pyridine-4,7-diamine in step 1) of the above Example 45, the same process as steps 1), 2) and 5) of the above Example 1 was performed sequentially to obtain 140 mg of the title compound (final yield: 57%).

¹ H-NMR (300 MHz, CDCl ₃ ): δ 9.40 (s, 1H), 8.62 (s, 1H), 8.28 (m, 1H), 8.08 (s, 1H), 8.03 (m, 4H), 7.75 (m, 1H), 7.29 (m, 3H), 6.99 (m, 1H), 6.64 (m, 1H), 4.12 (s, 3H), 2.39 (s, 3H).

Step 2) 3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6-difluoro- N -(5 H -Pyrrolo[2,3- b Preparation of ]pyrazin-2-yl)benzamide

70 mg (0.11 mmol) of the compound prepared in the above step 2) was diluted with 2 mL of a mixed solvent of methanol: tetrahydrofuran: distilled water (1: 1: 1 (volume ratio)), 43 mg of sodium hydroxide was added, and the mixture was stirred at room temperature for 3 hours. When the reaction was complete, the resulting reaction mixture was cooled to 0 °C, diluted with a mixed solvent of chloroform: 2-propanol (3: 1 (volume ratio)), and acidified with 1 N hydrochloric acid aqueous solution (pH 2). The resulting organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure. The resulting solid was diluted with dichloromethane: methanol = 9: 1 (volume ratio)), washed with dichloromethane, and filtered under reduced pressure. The resulting solid was dried under reduced pressure to obtain 34 mg (yield: 62%) of the title compound.

^1H -NMR (300MHz, DMSO- d ₆ ): δ 11.35 (s, 1H), 10.46 (s, 1H), 8.96 (s, 1H), 8.47 (s, 1H), 8.06 (m, 1H), 7.85 (m, 1H), 7.36 (m, 1H), 7.13 (m, 1H), 6.50 (m, 1H), 3.92 (s, 3H).

MS (ESI ⁺ ): m/z = 495.04 [M+H] ⁺ .

Example 49: N Preparation of -(3-(2-amino-5-fluoroquinazolin-6-yl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide

Step 1) 6-Bromo-5-fluoro- N Preparation of -(4-methoxybenzyl)quinazolin-2-amine

1.0 g (3.82 mmol) of 6-bromo-2-chloro-5-fluoroquinazoline was diluted in 10 mL of ethanol, 500 μL (3.82 mmol) of 4-methoxybenzylamine was added, and the mixture was stirred at 70 °C for 5 hours in a sealed tube. Upon completion of the reaction, the resulting reaction mixture was cooled to room temperature, and the resulting solid was washed with an aqueous ammonium chloride solution, filtered under reduced pressure, and dried under reduced pressure to obtain 1.1 g (yield: 76%) of the title compound.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 9.22 (s, 1H), 7.74 (m, 1H), 7.33 (m, 3H), 6.91 (m, 2H), 5.75 (m, 1H), 4.70 (m, 2H), 3.82 (s, 3H).

Step 2) 5-Fluoro- N Preparation of -(4-methoxybenzyl)-6-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)quinazolin-2-amine

1.1 g (2.9 mmol) of the compound prepared in the above step 1) was diluted in 30 mL of 1,4-dioxane, and 812 mg (3.2 mmol) of bis(pinacolato)diboron, 1.1 g (11.3 mmol) of potassium acetate, and 212 mg (0.29 mmol) of bis(triphenylphosphine)palladium(II) dichloride were added, and the mixture was stirred at 110°C for 4 hours. After the reaction was completed, the resulting reaction mixture was cooled to room temperature, diluted in ethyl acetate, washed with ethyl acetate through a Celite-packed filter, and filtered under reduced pressure, and the resulting filtrate was washed with distilled water. The resulting organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure. The resulting residue was separated by column chromatography (ethyl acetate: hexane = 1: 2 (volume ratio)) to obtain 660 mg (yield: 51%) of the title compound.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 9.27 (s, 1H), 7.95 (m, 1H), 7.36 (m, 3H), 6.92 (m, 2H), 5.83 (m, 1H), 4.72 (d, 2H), 3.82 (s, 3H), 1.29 (s, 12H).

Step 3) Preparation of 4-(pyridin-3-yl)pyrimidin-2-amine

200 mg (0.49 mmol) of the compound prepared in the above step 2) and 248 mg (0.54 mmol) of the compound prepared in the above example 1 step 8) were diluted with 4 mL of a mixed solvent of N , N -dimethylformamide:water (8:1 (volume ratio)), 156 mg (1.47 mmol) of sodium carbonate and 35 mg (0.049 mmol) of bis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium(II) were added, and the mixture was stirred at 110°C for 1 hour. After the reaction was completed, the resulting reaction mixture was cooled to room temperature, washed with ethyl acetate through a Celite-packed filter, filtered under reduced pressure, and the resulting filtrate was washed with distilled water. The resulting organic layer was dried over anhydrous sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure. The resulting residue was separated by column chromatography (ethyl acetate: hexane = 1:3 (volume ratio)) to obtain 53 mg (yield: 18%) of the title compound.

^1H -NMR (300MHz, DMSO- d ₆ ): δ 10.43 (s, 1H), 9.31 (s, 1H), 8.52 (d, 1H), 8.34 (m, 1H), 8.06 (d, 1H), 7.59 (t, 1H), 7.40 (m, 5H), 6.86 (d, 2H), 4.57 (d, 2H), 3.90 (s, 3H), 3.72 (s, 3H).

Step 4) Preparation of 2,4-difluoro-3-((trimethylsilyl)ethynyl)aniline

The same process as step 4) of Example 26 was performed except that the compound prepared in step 3) was used instead of N- (3-((4-amino-1-((2-(trimethylsilyl)ethoxy)methyl)-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide in step 4) of Example 26, to obtain 35 mg (yield: 82%) of the title compound.

^1H -NMR (300MHz, DMSO- d ₆ ): δ 10.44 (s, 1H), 9.31 (s, 1H), 8.52 (d, 1H), 8.07 (d, 1H), 7.60 (t, 1H), 7.44 (m, 1H), 7.26 (m, 4H), 3.90 (s, 3H).

MS (ESI ⁺ ): m/z = 496.04 [M+H] ⁺ .

The compounds of Examples 50 to 59 shown in Table 5 below were each prepared using the same or similar method as that of Example 49.

[Table 5]

Example 60: 5-chloro- N Preparation of -(2,4-difluoro-3-(quinazolin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide

Step 1) 5-chloro- N Preparation of -(2,4-difluoro-3-(quinazolin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide

Except that 2,4-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)aniline (WO2018049200) was used instead of 5-fluoro- N -(4-methoxybenzyl)-6-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)quinazolin-2-amine in step 3) of the above Example 49, and 6-bromoquinazoline was used instead of 5-chloro- N -(2,4-difluoro-3-iodophenyl)-2-methoxypyridine-3-sulfonamide, the same process as step 3) of Example 49 and step 8 of Example 1 was performed sequentially to obtain 64 mg of the title compound (final step yield: 60%).

^1H -NMR (300MHz, DMSO- d ₆ ): δ 10.47 (s, 1H), 9.70 (s, 1H), 9.38 (s, 1H), 8.52 (d, 1H), 8.22 (m, 1H), 8.16 (m, 1H), 8.09 (m, 1H), 8.00 (m, 1H), 7.40 (m, 1H), 7.29 (m, 4H), 3.92 (s, 3H).

MS (ESI ⁺ ): m/z = 463.04 [M+H] ⁺ .

The compounds of Examples 61 to 62 shown in Table 6 below were each prepared using the same or similar method as that of Example 60 above.

[Table 6]

Experimental Example 1: Evaluation of GCN2 kinase inhibitory activity

The GCN2 kinase inhibitory activity of the compounds prepared in Examples 1 to 62 was evaluated using a HotSpot kinase assay at Reaction Biology Corporation (Malbern, PA). Specifically, all compounds provided for testing were dissolved in DMSO (10 mM). Serial dilutions were performed by epMotion 5070 in DMSO. Specific enzyme/substrate mixtures and required cofactors were prepared in assay buffer (20 mM Hepes (pH 7.5), 10 mM MgCl ₂ , 1 mM EGTA, 0.02% Brij 35, 0.02 mg/ml BSA, 0.1 mM Na 3 VO 4 , 2 mM DTT, 1% DMSO).

Compounds were added to the enzyme reaction mixture using acoustic technology (Echo550; nanoliter range) and incubated at room temperature for 20 minutes. The enzyme reaction was initiated by adding 33P-ATP to a final concentration of 10 μM for single-point enzyme activity readings or to the ATP Km of GCN2 for IC ₅₀ readings. The reaction was incubated at room temperature for 2 hours, and enzyme activity was detected by the filter-binding method. IC ₅₀ values and curves were generated using GraphPad Prism 4.

[Table 7]

IC ₅₀ : <100 nM = A; 100-1,000 nM = B; >1,000 nM = C

As shown in Table 7 above, the compound of the present invention was shown to have excellent GCN2 kinase inhibitory activity.

Experimental Example 2: Evaluation of eIF2α kinase inhibitory activity

The eIF2α kinase inhibitory activity of the compounds in Table 8 below was evaluated at Reaction Biology Corporation (Malbern, PA) in the same manner as the GCN2 kinase inhibitory activity evaluation method of Experimental Example 1 above.

[Table 8]

IC ₅₀ : <100 nM = A; 100-1,000 nM = B; >1,000 nM = C

As shown in Table 8 above, the compound of the present invention was shown to have excellent eIF2α kinase inhibitory activity.

Experimental Example 3: Evaluation of eIF2α phosphorylation inhibitory activity

The eIF2a phosphorylation inhibitory activity of the compounds prepared in Examples 1 to 62 was evaluated in U2OS cell lines using the AlphaLISA® SureFire® Ultra ^TM assay. Specifically, according to the simple instructions in the assay kit manual of AlphaLISA® SureFire® Ultra ^TM , 3.3 X 10 ⁴ cells/well of U2OS cells were seeded in 96-well plates (Corning, 3599) in appropriate medium containing 10% FBS (Gibco, 16000044). The following day, the cells were treated with each compound (100 nM~, 1/5 dilution, 6 points) for 4 hours in amino acid-free RPMI1640 medium (USBiological Life Science, R8999-04A). Thereafter, the cells were washed with PBS and lysed with lysis buffer. AlphaLISA assay of phospho-eIF2a was performed according to a manual protocol, and alpha intensity was measured by a Synergy ^TM NEO HTS multimode microplate reader. Data were generated by nonlinear regression (Normalized Response) using GraphPad Prism V6 program.

[Table 9]

IC ₅₀ : <10 nM = A; 10-100 nM = B; >100 nM = C

As shown in Table 9 above, the compound of the present invention was shown to have excellent phosphorylation inhibition activity of eIF2α, a GCN2 downstream signal.

Hereinafter, the present invention will be described with reference to the above embodiments, but these are merely examples, and it should be understood that the present invention can be implemented within the scope of the appended claims by various modifications and other equivalent embodiments that are obvious to those skilled in the art.

The present invention relates to a novel nitrogen-containing heterocyclic derivative compound and its use, and more particularly, to a novel nitrogen-containing heterocyclic derivative having an activity inhibitory effect on eIF2α-activated kinases including GCN2, a pharmaceutically acceptable salt, optical isomer, hydrate or solvate thereof, a pharmaceutical composition comprising such a compound as an active ingredient, and its use, and can be usefully used as an agent for preventing or treating immune diseases including cancer or Alzheimer's disease by inhibiting the activity of eIF2α-activated kinases including GCN2.

Claims (12)

Any one compound selected from the compounds of the following formula I, pharmaceutically acceptable salts, optical isomers, hydrates and solvates thereof: [Chemical Formula I] In the above chemical formula I, Ring A is any one nitrogen-containing heterocycle selected from the group consisting of A-1 to A-5 below; In the above A-1 to A-5, Wavy lines ( ) indicates the attachment site; A plurality of X ⁰ are different from each other, one of which is C and the rest are N; A plurality of X ¹ are identical or different, and each independently is C(R) or N, or at least one is N; A plurality of X ² are different from each other, one of which is C(R') and the rest are N; X ³ is C(R'') or N; R and R'' are each independently hydrogen, halogen, C ₁ -C ₃ alkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R' is halogen, C ₁ -C ₃ alkyl, carboxamide (-CONH ₂ ), substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R ³ is hydrogen, or a substituted or unsubstituted amino group; The substituent of R ³ is a substituted or unsubstituted C ₁ -C ₆ alkyl, a substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, a substituted or unsubstituted 3 to 10 membered heterocycloalkyl, a substituted or unsubstituted C ₆ -C ₁₂ aryl, or a substituted or unsubstituted 5 to 12 membered heteroaryl; R ⁴ to R ⁷ are each independently hydrogen, halogen, substituted or unsubstituted C ₁ -C ₃ alkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; When each of R ⁴ to R ⁷ is alkyl, aryl or heteroaryl, their substituents may each independently be hydroxy, hydroxy C ₁ -C ₆ alkyl, or C ₁ -C ₆ alkoxy; R ⁸ is hydrogen or halogen; Ring B is arylene or heteroarylene substituted or unsubstituted with R ¹ ; R ¹ is each independently hydrogen, hydroxy, cyano, nitro, (mono-, di-, or trihalogen)methyl, halogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkene, C ₂ -C ₆ alkyne, C ₁ -C ₃ alkoxy, C ₃ -C ₆ cycloalkyl, or C ₁ -C ₆ dialkylamino; L ¹ is C _0- C ₆ alkylene, C _2- C ₆ alkenylene, C _2- C ₆ alkynylene, -NR ⁹ -CO-, -CO-NR ¹⁰ -, -NR ⁹ -CO-NR ¹⁰ -, -NR ⁹ -SO ₂ -, -NR ⁹ -SO ₂ -NR ¹⁰ -, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene, cycloalkylene or heterocycloalkylene, or may be a direct bond; L ² is -NR ⁹ -CO-, -CO-NR ¹⁰ -, -NR ⁹ -SO ₂ -, or -NR ⁹ -SO-NR ¹⁰ -; R ⁹ and R ¹⁰ are each independently hydrogen or C ₁ -C ₆ alkyl; Ring C is aryl or heteroaryl substituted or unsubstituted with R ² ; R ² can be independently selected from the group consisting of hydrogen, hydroxy, hydroxy C ₁ -C ₆ alkyl, cyano, C _1- C ₆ alkyl, C _2- C ₆ alkene, C _2- C ₆ alkyne, C _1- C ₃ alkoxy, (mono-, di-, or trihalogen)methyl, halogen, C _3- C ₆ cycloalkyl, C _1- C ₆ dialkylamino, substituted or unsubstituted aryloxy or heteroaryloxy, substituted or unsubstituted arylalkyloxy or heteroarylalkyloxy, substituted or unsubstituted aryl or heteroaryl, and substituted or unsubstituted cycloalkyl or heterocycloalkyl; p and q are 0 and 1 respectively, and p+q=1; m is an integer from 0 to 4; n is an integer from 0 to 5; When R ¹ or R ² is plural, they are each the same or different. In the first paragraph, A compound characterized in that the above A-1 is any one heteroaryl selected from the following group: Here, R ³ is as defined in Chemical Formula I. In the first paragraph, A compound characterized in that the above ring B is an arylene substituted or unsubstituted with a halogen. In the first paragraph, A compound characterized in that the ring C is phenyl or pyridine substituted or unsubstituted with R ² . In the first paragraph, A compound characterized in that L ¹ is a direct bond, C _2- C ₆ alkynylene or -NH-CO-. In the first paragraph, A compound characterized in that L ² is -NH-CO- or -NH-SO ₂ -. In the first paragraph, A compound characterized by being selected from the group consisting of the following compounds: N -(3-((8-aminoimidazo[1,2- a ]pyrazin-5-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; 5-Chloro- N -(3-((8-(cyclopropylamino)imidazo[1,2- a ]pyrazin-5-yl)ethynyl)-2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro- N -(3-((8-((1-ethyl-1 H -pyrazol-4-yl)amino)imidazo[1,2- a ]pyrazin-5-yl)ethynyl)-2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide; N -(3-((8-cyclopropylamino)imidazo[1,2- a ]pyrazin-5-yl)ethynyl)-2,4-difluorophenyl)-5-(fluoromethyl)-2-methoxypyridine-3-sulfonamide; N- (2,4-difluoro-3-(imidazo[1,2- a ]pyrazin-5-yl-ethynyl)phenyl)-5-(fluoromethyl)-2-methoxypyridine-3-sulfonamide; N -(3-((8-aminoimidazo[1,2- a ]pyrazin-5-yl)ethynyl)-2-fluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((8-aminoimidazo[1,2- a ]pyrazin-5-yl)ethynyl)-2,4-difluorophenyl)-5-(difluoromethyl)-2-methoxypyridine-3-sulfonamide; N -(3-((8-aminoimidazo[1,2- a ]pyrazin-5-yl)ethynyl)-2,4-difluorophenyl)-5-(fluoromethyl)-2-methoxypyridine-3-sulfonamide; N -(3-((8-aminoimidazo[1,2- a ]pyrazin-5-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxynicotinamide; N -(3-((4-aminopyrazolo[1,5- a ]pyrazin-7-yl)ethynyl)-2,4-difluoro)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((5-aminoimidazo[1,2- c ]pyrimidin-8-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N— (3-((8-amino[1,2,4]triazolo[4,3- a ]pyrazin-5-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((5-amino-[1,2,4]triazolo[4,3- c ]pyrimidin-8-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((2-amino-4-(pyridin-3-yl)pyrimidin-5-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((2-amino-4-phenylpyrimidin-5-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((2-amino-4-(3-ethoxyphenyl)pyrimidin-5-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((2-amino-4-(pyridin-4-yl)pyrimidin-5-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((2-amino-4-(thiazol-5-yl)pyrimidin-5-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((2-amino-4-(pyridin-3-yl)pyrimidin-5-yl)ethynyl)-2-fluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((2-amino-4-(pyridin-3-yl)pyrimidin-5-yl)ethynyl)-4-fluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((2-amino-4-(pyridin-3-yl)pyrimidin-5-yl)ethynyl)phenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((2-amino-4-(pyridin-3-yl)pyrimidin-5-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methylpyridine-3-sulfonamide; N -(3-((2-amino-4-(pyridin-3-yl)pyrimidin-5-yl)ethynyl)-2,4-difluorophenyl)pyridine-3-sulfonamide; N -(3-((2-amino-4-(pyridin-3-yl)pyrimidin-5-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxynicotinamide; N- (2-amino-4-(pyridin-3-yl)pyrimidin-5-yl)-3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6-difluorobenzamide; N -(3-((4-amino-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((4-amino-1-methyl-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((4-amino-2-methyl-2 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((4-amino-1-ethyl-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((4-amino-1-isopropyl-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((4-amino-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2-fluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((4-amino-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-4-fluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((4-amino-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-2-methoxy-5-(trifluoromethyl)pyridine-3-sulfonamide; N -(3-((4-amino-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-5-(difluoromethyl)-2-methoxypyridine-3-sulfonamide; N -(3-((4-amino-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-5-(fluoromethyl)-2-methoxypyridine-3-sulfonamide; N -(3-((4-amino-3-methyl-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((4-amino-3-methyl-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-2,5-dichloro-3-(hydroxymethyl)benzenesulfonamide; N -(3-((4-amino-3-methyl-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-2,5-dichloro-4-propoxybenzenesulfonamide; N -(3-((4-amino-1-(2-methoxyethyl)-3-methyl-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((4-amino-1-(2-hydroxyethyl)-3-methyl-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-((4-amino-1-(2-hydroxyethyl)-3-methyl-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-5-(fluoromethyl)-2-methoxypyridine-3-sulfonamide; N -(3-((3-chloro-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-5-(fluoromethyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro- N -(3-((3-chloro-1-(2-hydroxyethyl)-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide; N -(3-((3-chloro-1-(2-hydroxyethyl)-1 H -pyrazolo[4,3- c ]pyridin-7-yl)ethynyl)-2,4-difluorophenyl)-5-(fluoromethyl)-2-methoxypyridine-3-sulfonamide; N -(4-amino-1 H -pyrazolo[4,3- c ]pyridin-7-yl)-3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6-difluorobenzamide; N- (4-amino-1 H -pyrazolo[4,3- c ]pyridin-7-yl)-2,6-difluoro-3-((2-methoxy-5-(trifluoromethyl)pyridine)-3-sulfonamido)benzamide; 3-amino-6-((3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6-difluorophenyl)ethynyl)pyrazine-2-carboxamide; 3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6-difluoro- N -(5 H -pyrrolo[2,3- b ]pyrazin-2-yl)benzamide; N -(3-(2-amino-5-fluoroquinazolin-6-yl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N -(3-(2-amino-5-fluoroquinazolin-6-yl)-2,4-difluorophenyl)-5-(fluoromethyl)-2-methoxypyridine-3-sulfonamide; N- (2,4-difluoro-3-(5-fluoro-2(((1 r ,4 r )-4-hydroxycyclohexyl)amino)quinazolin-6-yl)phenyl)-5-(fluoromethyl)-2-methoxypyridine-3-sulfonamide; N -(3-(3-aminoisoquinolin-7-yl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N- (3-(3-aminoisoquinolin-7-yl)-2,4-difluorophenyl)-5-(fluoromethyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro- N -(3-(2-cyclopropylamino)-5-fluoroquinazolin-6-yl)-2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide; N -(3-(2-cyclopropylamino)-5-fluoroquinazolin-6-yl)-2,4-difluorophenyl)-5-(fluoromethyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro- N -(2,4-difluoro-3-(5-fluoro-2-(((1 r ,4 r )-4-hydroxycyclohexyl)amino)quinazolin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; N -(3-(2-amino-5-fluoroquinazolin-6-yl)-2,4-difluorophenyl)-5-chloro-2-methoxynicotinamide; N -(3-(2-amino-5-fluoroquinazolin-6-yl)-2,4-difluorophenyl)-5-chloronicotinamide; N -(3-(2-aminoquinazolin-6-yl)-2,4-difluorophenyl)-5-(fluoromethyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro- N -(2,4-difluoro-3-(quinazolin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; N- (2,4-dichloro-3-(isoquinolin-7-yl)phenyl)-5-(fluoromethyl)-2-methoxypyridine-3-sulfonamide; and N- (2,4-Difluoro-3-(quinazolin-6-yl)phenyl-5-(fluoromethyl)-2-methoxypyridine-3-sulfonamide. A pharmaceutical composition comprising a compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt, optical isomer, hydrate or solvate thereof, as an active ingredient. A pharmaceutical composition having an action of inhibiting abnormal activity of eIF2α in claim 8. A pharmaceutical composition having an action of inhibiting abnormal activity of one or more kinases selected from the group consisting of HRI, PKR, PERK and GCN2 in claim 8. A pharmaceutical composition for preventing or treating cancer, immune disease or Alzheimer's disease caused by abnormal activity of GCN2 in claim 8. A method for treating or preventing a disease caused by abnormal activity of GCN2, comprising administering to a person in need of treatment a therapeutically effective amount of a compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt, optical isomer, hydrate or solvate thereof.