Novel compounds and their uses

Abstract

This invention provides a novel compound and a pharmaceutical composition comprising the compound for the prevention or treatment of diseases associated with decreased SOX9 expression. The compound and the composition are effective in preventing or treating diseases associated with decreased SOX9 expression.

Description

Technical Field

[0001] This invention relates to a novel compound, or a stereoisomer thereof, a solvate, or a pharmaceutically acceptable salt thereof, and its use in the prevention or treatment of diseases associated with reduced SOX9 expression. Background Technology

[0002] Osteoarthritis (OA), also widely known as degenerative arthritis, is a disease affecting more than 7% of the global population (approximately 528 million people). Conventional treatments for OA primarily focus on joint replacement surgery and pain management using chondroprotective agents (such as hyaluronic acid) and anti-inflammatory drugs. However, due to population aging, the incidence of OA continues to rise globally. Therefore, there is an urgent need to develop drugs that can directly treat OA rather than simply relieve pain. One of the pathogenic factors of OA is an imbalance in the enzyme regulation of the anabolic and catabolitic pathways of articular cartilage proteins, leading to the destruction of articular cartilage tissue and progressive cartilage loss. To this end, current research is underway to develop disease-modifying osteoarthritis drugs (DMOADs) that target these regulatory factors to inhibit disease progression. The recently developed Lorecivivint (SM04690) is an example of a disease-modifying osteoarthritis drug.

[0003] SOX9 (SRY-box transcription factor 9) is a key atypical protein regulating the expression of extracellular matrix genes in chondrocytes. It is known to regulate the expression of extracellular matrix genes (such as collagen) that are crucial for articular cartilage. Particularly in osteoarthritis patients, decreased SOX9 expression and transcriptional activity have been observed, with SOX9 and its target proteins showing a significant decrease in expression levels with increasing disease severity. One regulatory mechanism of transcription factor activity is transcription factor aggregation. When aggregation is induced, the resulting transcription factor aggregates can regulate gene expression. In other words, when SOX9 transcription factor aggregation is induced, SOX9 expression levels and transcriptional effects can be enhanced through aggregate formation.

[0004] Therefore, there is an urgent need to develop disease-modifying osteoarthritis drugs (DMOADs) that can directly treat osteoarthritis by inducing the aggregation of SOX9 transcription factors and thereby improving their transcriptional efficiency. Summary of the Invention

[0005] Technical issues

[0006] One object of the present invention is to provide a novel compound, or a stereoisomer thereof, solvate, or pharmaceutically acceptable salt thereof, capable of inducing the aggregation of the SOX9 transcription factor.

[0007] Another object of the present invention is to provide a pharmaceutical composition that utilizes a novel compound or its stereoisomer, solvate or pharmaceutically acceptable salt capable of inducing SOX9 transcription factor aggregation for the prevention or treatment of diseases associated with reduced SOX9 expression.

[0008] Technical solution

[0009] The descriptions and embodiments disclosed in this specification can be applied to other descriptions and embodiments. That is, all combinations of the elements disclosed in this specification fall within the scope of protection of this invention. Furthermore, the scope of protection of this invention should not be limited by the detailed description below.

[0010] One aspect of the present invention provides a compound of Formula 1, or a stereoisomer, solvate, or pharmaceutically acceptable salt thereof: [Formula 1]

[0011] In Equation 1, Y 1 and Y 2 One of them is N, and the other is S, O, or NR. a ¹.

[0012] In Equation 1, Y 1 and Y 2 Each can be independently classified as O, S, or NR. a ².

[0013] In Equation 1, U is NR n2 , 5- to 7-membered heterocyclic groups containing 1 to 3 nitrogen atoms, or structural segments formed by the interconnection of these groups.

[0014] In Equation 1, Z 1 C 1-6 Alkylene.

[0015] In Equation 1, Z 2 For direct keys, C 1-6 Alkylene, -NR n3 CO-, or a 5- to 7-membered heteroaryl group containing 1 to 3 heteroatoms selected from N, O and S.

[0016] In Equation 1, R a1 and R a2 Each independently is H or C 1-6 alkyl.

[0017] In Equation 1, R n1 R n2 and R n3 Each independently is H or C 1-6 alkyl.

[0018] In Equation 1, when Z2 C 1-6 In the case of alkylene groups, any carbon atom of the alkylene group may optionally be converted by a halogen, a hydroxyl group, or a C-type carbon atom. 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 Alkyl, carboxyl, C 1-6 alkoxycarbonyl, carbamoyl, C 1-6 Alkyl carbamoyl, di(C 1-6 Alkyl) carbamoyl, cyano, nitro, oxo, or C 6-12 Aryl substitution.

[0019] In Equation 1, Z 3 For direct keys or -C(=O)-.

[0020] In Formula 1, ring A is a 6- to 14-membered aryl group, a partially unsaturated 9- to 14-membered bicyclic carbocyclic group, a 5- to 12-membered heteroaryl group containing 1 to 3 heteroatoms selected from O, N or S, a 5- to 8-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N or S, or a fused benzo[a]heterocyclic group formed by fusion of a benzene ring with a 5- to 7-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N or S.

[0021] In Equation 1, ring A is arbitrarily divided by 1 to 3 R. A replace.

[0022] R A Choose free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 The group consisting of alkyl, cyano, nitro and oxo groups.

[0023] In Formula 1, ring E is a 6- to 14-membered aryl group, a partially unsaturated 9- to 14-membered bicyclic carbocyclic group, a 5- to 12-membered heteroaryl group containing 1 to 3 heteroatoms selected from O, N, or S, a 5- to 12-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N, or S, or a fused heterocyclic aryl group formed by fusion of a 5- to 7-membered heterocycle containing 1 to 3 heteroatoms selected from O, N, or S with a phenyl group.

[0024] In Equation 1, ring E is arbitrarily divided by 1 to 3 R. E replace.

[0025] R E Choose free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, carboxyl, C 1-6 alkoxycarbonyl, carbamoyl, C1-6 Alkyl carbamoyl, di(C 1-6 Alkyl) carbamoyl, halosulfonyl, sulfinyl, C 1-6 alkylsulfonyl, cyano, nitro, oxo, or optionally substituted with one or more groups selected from halogen, hydroxyl, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Substituents of alkyl, amino, nitro and cyano groups on C 1-6 A group composed of alkyl groups.

[0026] In Equation 1, Y 1 and Y 2 One of them can be N, and the other can be S, O, or NR. a ¹. For example, Y 1 It can be N, Y 2 Can be S, O or NR a ¹; or Y 2 It can be N, Y 1 Can be S, O or NR a ¹. In some implementations, Y 1 and Y 2 One of them can be N, and the other can be S or O. For example, Y 1 It can be N, and Y 2 It can be S; or Y 1 It can be S or O, and Y 2 It can be N.

[0027] In Equation 1, X 1 and X 2 They can be independently designated as O, S, or NR. a ². X 1 and X 2 They can be the same or different. For example, X 1 and X 2 Each can be O.

[0028] In Equation 1, U can be NR n2 A 5- to 7-membered heterocyclic group containing 1 to 3 nitrogen atoms, or a structural fragment formed by the interconnection of these groups. In some embodiments, U may be NR. n2 , 5 or 6-membered heterocyclic groups containing 1 or 2 nitrogen atoms, or structural segments formed by the interconnection of these groups.

[0029] In some implementations, U can be selected from NR. n2 Piperidine dimethyl, piperazine dimethyl, or NR n2 Structural segments formed by linking piperidine dimethyl or piperazine dimethyl.

[0030] In some implementations, U can be selected from NR.n2 A group consisting of the following structures. In each of the following structures, 1 It can be connected to -C(=X) in Equation 1 2 carbon atoms of the )- 2 Can be connected to Z 2 .

[0031]

[0032] R a1 and R a2 Each can be H or C independently. 1-6 Alkyl group. In some embodiments, R a1 and R a2 Each can be H or C independently. 1-4 Alkyl group. For example, R a1 and R a2 Each can be either H or methyl.

[0033] In Equation 1, R n1 R n2 and R n3 Each can be H or C independently. 1-6 Alkyl group. In some embodiments, R n1 R n2 and R n3 Each can be H or C independently. 1-4 Alkyl group. For example, R n1 R n2 and R n3 Each can be either H or methyl.

[0034] In Equation 1, Z 1 It can be C 1-6 Alkylene. In some embodiments, Z 1 It can be C 1-4 Alkylene. In some embodiments, Z 1 It can be a straight-chain alkylene group. In some embodiments, Z 1 It can be methylene, ethylene, propylene, butylene, pentylene, or hexylene.

[0035] In Equation 1, Z 2 Can be a direct key, C 1-6 Alkylene, -NR n3 CO- or a 5- to 7-membered heteroaryl group containing 1 to 3 heteroatoms selected from N, O, and S. -NR n3 CO- can be 3 -NR n3 CO- 4 ,in 3 It can be connected to U in Equation 1. 4 It can be connected to ring E. In some implementations, Z 2 Can be a direct key, C 1-4 Alkylene, -NR n3 CO- or a 5- or 6-membered heteroaryl group containing 1 to 3 heteroatoms selected from N, O, and S. For example, Z 2 It can be a direct bond, methylene, ethylene, propylene, butylene, -NHCO-, or dithiazole dimethyl.

[0036] In Equation 1, when Z 2 C 1-6 In the case of alkylene groups, any carbon atom of the alkylene group may optionally be replaced by a halogen, a hydroxyl group, or a C-type carbon atom. 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 Alkyl, carboxyl, C 1-6 alkoxycarbonyl, carbamoyl, C 1-6 Alkyl carbamoyl, di(C 1-6 Alkyl) carbamoyl, cyano, nitro, oxo, or C 6-12 Aryl substitution. In some embodiments, any carbon atom of the alkylene group may optionally be replaced by a carboxyl group, C... 1-6 alkoxycarbonyl or C 6-12 Aryl substitution. For example, any carbon atom of the alkylene group may be substituted with a methoxycarbonyl group or a phenyl group. 2 The carbon atom adjacent to ring E in the alkylene group can be substituted.

[0037] In some implementations, U is a 5- to 7-membered heterocyclic group containing 1 to 3 N heteroatoms, or NR n2 When Z is linked with structural segments consisting of 5- to 7-membered heterocyclic groups containing 1 to 3 N heteroatoms, 2 Can be a direct key or C 1-6 Alkylene.

[0038] In Equation 1, Z 3 It can be a direct key or -C(=O)-.

[0039] In some embodiments, the compound shown in Formula 1 may be selected from the compounds shown in Formula 2 or Formula 3 below.

[0040] [Equation 2]

[0041] [Formula 3]

[0042] In equations 2 and 3, Z 1 Z 2 X 1 X 2 ,U,R n1 R n2 Ring A and ring E are defined as in Equation 1.

[0043] In some implementations, Y in Equation 2 1 Y in formula 3 2 Each can be either S or O independently.

[0044] In Formula 1, ring A may be a 6- to 14-membered aryl group, a partially unsaturated 9- to 14-membered bicyclic carbocyclic group, a 5- to 12-membered heteroaryl group containing 1 to 3 heteroatoms selected from O, N, or S, a 5- to 8-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N, or S, or a fused benzo[a]heterocyclic group formed by fusion of a benzene ring with a 5- to 7-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N, or S. In some embodiments, ring A may be a 6- to 12-membered aryl group, a partially unsaturated 9- or 10-membered bicyclic carbocyclic group, a 6- to 10-membered heteroaryl group containing 1 or 2 heteroatoms selected from O, N, or S, a 5- to 7-membered heterocyclic group containing 1 or 2 heteroatoms selected from O, N, or S, or a fused benzo[a]heterocyclic group formed by fusion of a benzene ring with a 5- to 7-membered heterocyclic group containing 1 or 2 heteroatoms selected from O, N, or S.

[0045] In some embodiments, ring A may be phenyl, biphenyl, tetrahydronaphthyl, pyranyl, pyranone (oxopyranyl), benzopyranyl, or benzopyranone. The benzopyranone group may be coumarinyl.

[0046] In some implementations, ring A can be , ,or Furthermore, ring A can be arbitrarily divided by 1 to 3 R's. A replace.

[0047] In some implementation schemes, Z 3 It can be a direct bond, and ring A can be... , ,or ;or

[0048] Z 3 It can be -C(=O)-, and ring A can be or .

[0049] In Equation 1, ring A can be optionally divided by 1 to 3 R. A Replace. R A Optional free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 The group consisting of alkyl, cyano, nitro, and oxo groups. In some embodiments, R A Optional free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 The group consisting of alkyl and oxo groups.

[0050] In some implementation schemes, R A Optional free halogens, hydroxyl groups, C 1-4 Alkoxy, amino, C 1-4 Alkylamino, di(C) 1-4 Alkyl)amino, C 1-4 The group consisting of alkyl, cyano, nitro and oxo groups.

[0051] In some implementation schemes, R A The group consisting of F, Cl, Br, hydroxyl, methoxy, dimethylamino, methyl, and oxo can be selected.

[0052] In some implementations, ring A may be selected from the following chemical structures: .

[0053] In Formula 1, ring E may be a 6- to 14-membered aryl group, a partially unsaturated 9- to 14-membered bicyclic carbocyclic group, a 5- to 12-membered heteroaryl group containing 1 to 3 heteroatoms selected from O, N, or S, a 5- to 12-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N, or S, or a fused heterocyclic aryl group formed by fusion of a 5- to 7-membered heterocycle containing 1 to 3 heteroatoms selected from O, N, or S with a phenyl group. In some embodiments, ring E may be a 6- to 12-membered aryl group, a partially unsaturated 9- to 12-membered bicyclic carbocyclic group, a 5- to 12-membered heteroaryl group containing 1 or 2 heteroatoms selected from O, N, or S, a 5- to 8-membered heterocyclic group containing 1 or 2 heteroatoms selected from O, N, or S, or a fused heterocyclic aryl group formed by fusion of a 5- to 7-membered heterocycle containing 1 or 2 heteroatoms selected from O, N, or S with a phenyl group. In some embodiments, ring E may be a 6- to 10-membered aryl group, a partially unsaturated 9- or 10-membered bicyclic carbocyclic group, a 5- to 10-membered heteroaryl group containing one or two heteroatoms selected from O, N, or S, a 5- to 7-membered heterocyclic group containing one or two heteroatoms selected from O, N, or S, or a fused heterocyclic aryl group formed by fusion of a 5- or 6-membered heterocycle containing one or two heteroatoms selected from O, N, or S with a phenyl group.

[0054] In some embodiments, ring E may be phenyl, naphthyl, indanyl, pyridinyl, pyridinyl, imidazole, pyrazolyl, triazolyl, thiophene, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, morpholinyl, thiomorpholinyl, piperidinyl, hexahydroxypyridinyl, hexahydroxypyrimidinyl, piperazinyl, indole, inzolyl, benzimidazolyl, quinolinyl, isoquinolinyl, cenyl, quinazolinyl, quinoxalinyl, phthalazinyl, or benzodioxolane. For example, ring E may be phenyl, naphthyl, pyridinyl, pyridinyl, imidazole, thiophene, morpholinyl, thiomorpholinyl, piperazinyl, indole, quinolinyl, or benzodioxolane.

[0055] In some embodiments, ring E may be selected from the chemical structures shown below; and ring E may optionally be surrounded by 1 to 3 R groups. E replace.

[0056] .

[0057] In Equation 1, the ring E can be optionally divided by 1 to 3 R. E Replacement. When there are two or more R... E At that time, each R E They may be the same as or different from each other.

[0058] R E Optional free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, carboxyl, C 1-6 alkoxycarbonyl, carbamoyl, C 1-6 Alkyl carbamoyl, di(C 1-6 Alkyl) carbamoyl, halosulfonyl, sulfinyl, C 1-6 alkylsulfonyl, cyano, nitro, oxo, or optionally substituted with one or more groups selected from halogen, hydroxyl, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Substituents of alkyl, amino, nitro and cyano groups on C 1-6 The group consisting of alkyl groups; in some embodiments, R E Choose free halogens, hydroxyl groups, C 1-4 Alkoxy, amino, C 1-4 Alkylamino, di(C) 1-4 Alkyl)amino, carboxyl, C 1-4 alkoxycarbonyl, carbamoyl, C 1-4 Alkyl carbamoyl, di(C 1-4 Alkyl) carbamoyl, halosulfonyl, sulfinyl, C1-6 alkylsulfonyl, cyano, nitro, oxo, or optionally substituted with one or more groups selected from halogen, hydroxyl, C 1-4 Alkoxy, amino, C 1-4 Alkylamino, di(C) 1-4 Substituents of alkyl, amino, nitro and cyano groups on C 1-4 Groups composed of alkyl groups; In some implementation schemes, R E Optional free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 alkyl)amino, halosulfonyl, sulfinyl, C 1-6 Alkyl sulfonyl, cyano, nitro, oxo, C 1-6 Alkyl, C 1-6 Halogenated alkyl groups and C 1-6 The group consisting of hydroxyalkyl groups. In some embodiments, R E Optional free halogens, hydroxyl groups, C 1-4 Alkoxy, amino, C 1-4 Alkylamino, di(C) 1-4 alkyl)amino, halosulfonyl, sulfinyl, C 1-4 Alkyl sulfonyl, cyano, nitro, oxo, C 1-4 Alkyl, C 1-4 Halogenated alkyl groups and C 1-4 The group consisting of hydroxyalkyl groups.

[0059] In some implementation schemes, R E The group consisting of free methoxy, hydroxy, amino, F, Cl, Br, trifluoromethyl, cyano, hydroxymethyl, methyl, fluorosulfonyl, nitro and oxo groups can be selected.

[0060] In some implementations, ring E may be selected from the following chemical structures: .

[0061] In some embodiments, the compound represented by Formula 1 may be represented by Formulas I to III.

[0062] [Formula I]

[0063] In equation I, Y 1 For S, O or NR a1 ;R a1 For H or C 1-6Alkyl; R n1 With R n2 Each independently is H or C 1-6 Alkyl; Z 4 For direct key or C 1-4 Alkylene; R 1 H, halogen, hydroxyl, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 Alkyl, carboxyl, C 1-6 alkoxycarbonyl, carbamoyl, C 1-6 Alkyl carbamoyl, di(C 1-6 Alkyl) carbamoyl, cyano, nitro, oxo or C 6-12 Aryl.

[0064] In Formula I, ring A is a 6- to 14-membered aryl group, a partially unsaturated 9- to 14-membered bicyclic carbocyclic group, a 6- to 12-membered heteroaryl group containing 1 to 3 heteroatoms selected from O, N, or S, a 5- to 8-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N, or S, or a fused benzo[a]heterocyclic group formed by fusion of a benzene ring with a 5- to 7-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N, or S; ring A is optionally surrounded by 1 to 3 R[a] A Replace; and R A Optional free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 The group consisting of alkyl, cyano, nitro and oxo groups.

[0065] In Formula I, ring E is a 6- to 14-membered aryl group, a partially unsaturated 9- to 14-membered bicyclic carbocyclic group, a 5- to 12-membered heteroaryl group containing 1 to 3 heteroatoms selected from O, N, or S, a 5- to 12-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N, or S, or a fused heterocyclic aryl group formed by fusion of a 5- to 7-membered heterocycle containing 1 to 3 heteroatoms selected from O, N, or S with a phenyl group; ring E is optionally surrounded by 1 to 3 R groups. E Replace; and R E Optional free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, carboxyl, C 1-6 alkoxycarbonyl, carbamoyl, C 1-6 Alkyl carbamoyl, di(C 1-6 Alkyl) carbamoyl, halosulfonyl, sulfinyl, C 1-6 alkylsulfonyl, cyano, nitro, oxo, or optionally substituted with one or more groups selected from halogen, hydroxyl, C1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Substituents of alkyl, amino, nitro and cyano groups on C 1-6 A group composed of alkyl groups.

[0066] [Formula II]

[0067] In Equation II, Y 2 For S, O or NR a1 ;R a1 For H or C 1-6 Alkyl; R n1 With R n2 Each independently is H or C 1-6 Alkyl; Z 4 For direct key or C 1-4 Alkylene; R 1 H, halogen, hydroxyl, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 Alkyl, carboxyl, C 1-6 alkoxycarbonyl, carbamoyl, C 1-6 Alkyl carbamoyl, di(C 1-6 Alkyl) carbamoyl, cyano, nitro, oxo or C 6-12 Aryl.

[0068] In Equation II, ring A 1 It is a 6- to 14-membered aryl group or a partially unsaturated 9- to 14-membered bicyclic carbocyclic group; Ring A 1 Optionally by 1 to 3 R A Replace; and R A Optional free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 The group consisting of alkyl, cyano, nitro, and oxo groups. In some embodiments, cyclic A... 1 It can be 6 to 14 aryl groups.

[0069] In Formula II, ring E is a 6- to 14-membered aryl group, a 5- to 12-membered heteroaryl group containing 1 to 3 heteroatoms selected from O, N, or S, a 5- to 12-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N, or S, or a fused heterocyclic aryl group formed by fusion of a 5- to 7-membered heterocycle containing 1 to 3 heteroatoms selected from O, N, or S with a phenyl group; ring E is optionally surrounded by 1 to 3 R groups. E Replace; and R EChoose free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 Alkyl, carboxyl, C 1-6 alkoxycarbonyl, carbamoyl, C 1-6 Alkyl carbamoyl, di(C 1-6 Alkyl) carbamoyl, halosulfonyl, sulfinyl, C 1-6 The group consisting of alkylsulfonyl, cyano, nitro and oxo groups.

[0070] In some embodiments, the compound represented by Formula 1 may be selected from compounds represented by Formula IA, Formula IB, Formula IC, Formula ID, Formula IE or Formula IIA.

[0071] [Formula III]

[0072] In Equation III, U 1 For direct bond or NR n2 ;Y 3 and Y 4 At least one of them is N, and the other is CH; Z 5 For direct keys, C 1-6 Alkylene, -NR n3 CO-, or a 5- to 7-membered heteroaryl group containing 1 to 3 heteroatoms selected from N, O, and S; R n1 R n2 and R n3 Each independently is H or C 1-6 Alkyl group; and s is 0 or 1.

[0073] In Formula III, ring A may be a 6- to 14-membered aryl group, a partially unsaturated 9- to 14-membered bicyclic carbocyclic group, a 5- to 12-membered heteroaryl group containing 1 to 3 heteroatoms selected from O, N or S, a 5- to 8-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N or S, or a fused benzo[a]heterocyclic group formed by fusion of a benzene ring with a 5- to 7-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N or S.

[0074] In Equation III, ring A is optionally divided by 1 to 3 R. A Replace; and R A Optional free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 The group consisting of alkyl, cyano, nitro and oxo groups.

[0075] In Formula III, ring E may be a 6- to 14-membered aryl group, a partially unsaturated 9- to 14-membered bicyclic carbocyclic group, a 5- to 12-membered heteroaryl group containing 1 to 3 heteroatoms selected from O, N, or S, a 5- to 12-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N, or S, or a fused heterocyclic aryl group formed by fusion of a 5- to 7-membered heterocycle containing 1 to 3 heteroatoms selected from O, N, or S with a phenyl group.

[0076] In Equation III, ring A is optionally divided by 1 to 3 R. A Replace; and R E Optional free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, carboxyl, C 1-6 alkoxycarbonyl, carbamoyl, C 1-6 Alkyl carbamoyl, di(C 1-6 Alkyl) carbamoyl, halosulfonyl, sulfinyl, C 1-6 alkylsulfonyl, cyano, nitro, oxo, or optionally substituted with one or more groups selected from halogen, hydroxyl, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Substituents of alkyl, amino, nitro and cyano groups on C 1-6 Groups composed of alkyl groups; [Form IA]

[0077] [Form IB]

[0078] [Form IC]

[0079] [Formula ID]

[0080] [Formula IE]

[0081] [Form IIA]

[0082] In formulas IA, IB, IC, ID, IE, and IIA, R A Choose free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6The group consisting of alkyl, cyano, nitro, and oxo groups; n, m1, m2, o1, o2, p, and q are each independent integers from 0 to 3; the sum of m1 and m2 and the sum of o1 and o2 are each 3 or less; and Y 1 Y 2 R n1 R n2 Z 4 R 1 And ring E is as described in Equations I and II above.

[0083] In some embodiments, the compound represented by Formula 1 may be selected from compounds represented by the following chemical structures: .

[0084] In some embodiments, the compound represented by Formula 1 can be synthesized according to reaction scheme 1 below.

[0085] [Reaction Scheme 1]

[0086] In reaction scheme 1, Y 1 Y 2 Z 3 Ring A, X 1 X 2 Z 1 Z 2 R 1n U and ring E are as defined in Equation 1 above.

[0087] Specifically, in step 1 of reaction scheme 1, intermediate a, containing a five-membered heteroaromatic ring substituted with an amino group and optionally linked to ring A via Z3, can be reacted with cyclic intermediate 1 to synthesize intermediate b. The reaction in step 1 can be carried out in a suitable organic solvent capable of dissolving intermediate a or intermediate i, or in the molten state without organic solvent, at a temperature equal to or higher than the melting point of intermediate i (120°C). For example, the reaction in step 1 can be carried out at approximately 120°C to 180°C for 1 to 5 hours.

[0088] In step 2 of reaction scheme 1, intermediate b can be reacted with amine intermediate c containing cyclic E to prepare the compound represented by formula 1. Step 2 can be carried out under anhydrous conditions and can use an organic solvent (e.g., dioxane) at a temperature of about 80°C to 140°C for 5 to 24 hours.

[0089] Intermediate a can be synthesized according to reaction scheme 2 below.

[0090] [Reaction Scheme 2]

[0091] In reaction scheme 2, rings A and Z 3 As defined in Equation 1 above.

[0092] Specifically, in step 1 of reaction scheme 2, an acetyl compound containing ring A can be reacted with excess tetrabutylammonium tribromide (CAS: 38932-80-8) in an organic solvent to synthesize a brominated acetyl compound. The organic solvent used in this reaction can be, but is not limited to, CH2Cl2, and the reaction temperature can be from about 10°C to 50°C, for example, room temperature.

[0093] In step 2 of reaction scheme 2, the product of step 1 can be heated with thiourea in ethanol to prepare intermediate a'. This intermediate a' can be used as intermediate a in reaction scheme 1 above.

[0094] As used herein, the term "halogen" or "halogen atom" refers to an atom in Group 17 of the periodic table. Halogen atoms include F, Cl, Br, I, etc. The term "halogenated" refers to a halogen substituent.

[0095] The term "alkyl" refers to a fully saturated branched or unbranched (or straight-chain or linear) hydrocarbon group. The alkyl group may be substituted or unsubstituted. C 1-20 Alkyl groups, for example, can be C10. 1-15 C 1-10 Or C 1-6 Alkyl group. C 1-6 Alkyl groups can be C 1-5 C 1-4 C 1-3 Or C 1-2Alkyl groups. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, neopentyl, and n-hexyl.

[0096] The term "haloalkyl" refers to an alkyl group that has been substituted with one or more halogen atoms, and includes dihaloalkyl, trihaloalkyl, etc. "Haloalkyl" can also include perhaloalkyl, in which all hydrogen atoms of the alkyl group are substituted with halogen atoms.

[0097] The term "hydroxyl group" refers to the -OH functional group (hydroxyl group).

[0098] The term "hydroxyalkyl" refers to an alkyl group that has been substituted with a hydroxyl group.

[0099] The term "carbonyl" refers to -C(=O)-.

[0100] The term "alkoxy" refers to an alkyl group bonded to an oxygen atom. 1-20 Alkoxy groups can be, for example, C 1-15 C 1-10 Or C 1-6 Alkyl group. C 1-6 Alkoxy groups can be C 1-5 C 1-4 C 1-3 Or C 1-2 Alkyl groups. Examples of alkoxy groups include methoxy, ethoxy, propoxy, and butoxy.

[0101] The term "alkoxyalkyl" refers to an alkoxy group bonded to an alkyl group. C 2-20 Alkoxyalkyl groups, for example, can be C 2-15 C 2-10 Or C 2-6 Alkoxyalkyl. For example, C 2-20 Alkoxyalkyl groups can be (C 1-10 alkoxy)-(C 1-10 alkyl) or (C 1-6 alkoxy)-(C 1-6 Alkyl group). The alkoxy group may have the same or different number of carbon atoms as the alkyl group. Examples of alkoxyalkyl groups include methoxymethyl, methoxyethyl, ethoxymethyl, ethoxyethyl, methoxypropyl, and ethoxypropyl.

[0102] The term "amino" refers to -NH2.

[0103] The term "amine" refers to a substituent formed by replacing one, two, or all three hydrogen atoms of ammonia with an organic functional group. It includes primary, secondary, and tertiary amines and is a term that covers amino groups.

[0104] The term "alkylamine" refers to an amine formed by replacing one hydrogen atom of an amino group (-NH2) with an alkyl group.

[0105] The term "di(alkyl)amine" refers to an amine formed by replacing both hydrogen atoms in the amino group (-NH2) with alkyl groups. The two alkyl groups in a di(alkyl)amine may be the same or different.

[0106] The term "nitro" refers to -NO2.

[0107] The term "cyano" refers to -CN, a functional group formed by a carbon atom and a nitrogen atom connected by a triple bond.

[0108] The term "carboxyl group" refers to -COOH. Salts of carboxyl groups are the conjugate bases of carboxylic acids.

[0109] The term "alkoxycarbonyl" refers to a monovalent substituent formed when the -OH group in a carboxyl group is replaced by an alkoxy group. For example, C 1-6 Alkoxycarbonyl refers to the group formed by the carbonyl group (C). 1-6 Alkoxy-substituted -C(=O)-.

[0110] The term "carbamoyl" refers to -CONH2.

[0111] The term "alkylcarbamoyl" refers to a substituent formed when one hydrogen atom of the -NH2 group in carbamoyl is replaced by an alkyl group.

[0112] The term "dialkylcarbamoyl" refers to a substituent formed by replacing each of the two hydrogen atoms of the -NH2 group in a carbamoyl group with an alkyl group. In a dialkylcarbamoyl group, the two alkyl groups may be the same or different.

[0113] The term "sulfonyl" refers to the -SO2- group.

[0114] The term "halosulfonyl" refers to a sulfonyl group that has been substituted with a halogen, and it is a monovalent substituent.

[0115] The term "carbocyclic group" refers to a monovalent non-aromatic hydrocarbon ring substituent. The carbocyclic group can be fully saturated or partially unsaturated, and can have a monocyclic structure, or a fused, bridged, or spirocyclic bicyclic or tricyclic structure. For example, the carbocyclic group includes a group fused with a phenyl group and a saturated six-membered heterocycle, such as tetrahydronaphthyl. The carbocyclic group can be a substituent forming a ring of 9 to 14 carbon atoms.

[0116] The term "cycloalkyl" refers to a saturated non-aromatic monocyclic, bicyclic, or tricyclic hydrocarbon group. The cycloalkyl group may contain 3 to 20 carbon atoms, for example, 5 to 10, 3 to 8, or 3 to 6. Examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Examples of bicyclic cycloalkyl groups include borneol, decahydronaphthyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, and bicyclo[2.2.2]octyl. Examples of tricyclic cycloalkyl groups include adamantyl.

[0117] The term "cycloalkane" refers to a saturated, non-aromatic monocyclic, bicyclic, or tricyclic hydrocarbon ring. Cycloalkane rings can be in the complete (non-radical) form of cycloalkyl groups. They can contain 3 to 20 carbon atoms, for example, 5 to 10, 3 to 8, or 3 to 6. Examples of monocyclic cycloalkane rings include cyclopropane, cyclobutane, cyclopentane, and cyclohexane.

[0118] The term "aryl" refers to an aromatic hydrocarbon cyclic group. An aryl group comprises multiple aryl groups linked together to provide aromaticity. 6-30 Aryl groups can be C, for example. 6-14 C 6-12 Or C 6-10 Aryl 。 The aryl group may have a monocyclic structure, or a fused, bridged, or spirocyclic bicyclic or tricyclic structure. The aryl group may be phenyl, naphthyl, or biphenyl.

[0119] The term "arylalkyl" refers to an alkyl group that has been substituted with an aryl group.

[0120] The term "aryloxy group" refers to an aryl group bonded to an oxygen atom.

[0121] The term "heteroaryl" or "heteroaromatic hydrocarbon" refers to a monocyclic or bicyclic aromatic compound or substituent containing one or more heteroatoms and with the remaining ring atoms being carbon. The heteroaryl, even if its ring is partially unsaturated, may include cyclic groups that are aromatic as a whole through substitution of unsaturated ring atoms by oxo (=O), thio (=S), etc. The heteroaryl may contain, for example, 1 to 5, 1 to 3, or 1 or 2 heteroatoms and may contain 5 to 12 ring members. Examples of "heteroaryl" compounds include pyrroleyl, furanyl, thiophenyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrazinyl, pyridazinyl, indolyl, benzofuranyl, benzothiophenyl, benzopyrazolyl, benzoimidazolyl, benzooxazolyl, benzoisooxazolyl, benzothiazolyl, benzoisothiazolyl, oxopyranyl (pyranoneyl), benzopyranoneyl, and coumarinyl.

[0122] The term "heterocyclic alkyl" or "heterocyclic group" refers to a saturated or partially unsaturated cyclic hydrocarbon group containing at least one heteroatom. The heterocyclic cyclic group can be a monocyclic, bicyclic, or tricyclic group. The bicyclic group can be a spirocyclic, bridged, or fused ring group. The heterocyclic cyclic group can contain 3 to 20, 3 to 10, 3 to 8, 3 to 7, 5 to 7, 4 to 6, or 5 to 6 ring atoms. The heteroatom can be one or more atoms selected from the group consisting of N, O, and S, for example, 1, 2, or 3 heteroatoms. Examples of heterocyclic groups include pyranyl, aziridinyl, ethylene oxide, oxacyclobutyl, aziridine, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, piperidinyl, tetrahydropyranyl, dihydropyranyl, morpholinyl, thiomorpholinyl, piperazine, and oxazolyl.

[0123] The term "fused benzo[a]heterocyclic group" refers to a substituent formed by the fusion of a benzene ring and a heterocyclic group. The fused benzo[a]heterocyclic group can be a fused heterobicyclic group shared by two adjacent carbon atoms of the heterocyclic ring and the benzene ring. The fused benzo[a]heterocyclic group can also be a 9- to 11-membered fused heterobicyclic group formed by the fusion of a benzene ring and a 5- to 7-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N, and S. For example, the fused benzo[a]heterocyclic group can include a benzo[a]pyranyl group formed by the fusion of a benzene ring and a pyranyl group.

[0124] The term "fused heterocyclic aryl" refers to a substituent in which a heterocycle and an aryl group are fused. The fused heterocyclic aryl can be a fused heterobicyclic group shared by two adjacent carbon atoms of the heterocycle and an aryl group. The fused heterocyclic aryl can also be a 9- to 7-membered fused heterobicyclic group formed by the fusion of a 5- to 7-membered heterocycle containing 1 to 3 heteroatoms selected from O, N, and S with an aryl group. For example, the fused heterocyclic aryl can include a benzodioxolane group formed by the fusion of a dioxolane and a phenyl group.

[0125] The heteroatom may be one or more selected from the group consisting of N, O, P, and S. The heteroatom may be one, two, or three heteroatoms selected from the group consisting of N, O, and S.

[0126] In the terms “substituted or unsubstituted,” “substitution” means that when one or more hydrogen atoms in an organic compound are substituted by another group of atoms to form a derivative, that group of atoms is introduced to replace the hydrogen atoms. “Substituent” refers to the introduced group of atoms. As used herein, “substitution” without limiting the substituent can mean substitution by, for example, groups such as: halogen atoms, C1-C atoms substituted with halogen atoms. 20 Alkyl groups (e.g., CCF3, CHCF2, CH2F, CCl3, etc.), C1-C 20 Alkoxy, C2-C 20 Alkoxyalkyl, hydroxyl, -NH2, =NH, nitro, cyano, amidine, hydrazine, hydrazone, carboxyl or its salt, sulfonyl, aminosulfonyl, sulfonic acid or its salt, phosphoric acid or its salt, C1-C 20 Alkyl, C2-C 20 alkenyl, C2-C 20 alkynyl group, C6-C 20 Aryl, C6-C 20 arylalkyl, C6-C 20 heteroaryl, C7-C 20 Heteroarylalkyl, C6-C 20 Heteroaryloxy, C6-C 20 Heteroaryloxyalkyl or C6-C 20 Heteroarylalkyl.

[0127] The terms "stereoisomer" or "isomer" refer to compounds with the same molecular formula but different atomic arrangements or spatial configurations. Isomers include, for example, structural isomers and stereoisomers. Stereoisomers can be diastereomers or enantiomers. Enantiomers are isomers that are mirror images of each other, like the left and right hands; they are also called optical isomers. When the four or more substituents on the chiral central carbon atom are different from each other, enantiomers are designated as R (Rectus: clockwise) and S (Sinister: counterclockwise). Diastereomers are stereoisomers that are not mirror images of each other, arising from differences in atomic spatial arrangement. Diastereomers can be classified into cis-trans isomers and conformational isomers (conformations).

[0128] The term "solvent" refers to a compound that has been solvated by an organic or inorganic solvent. A solvate is, for example, a hydrate.

[0129] The term "salt" refers to both inorganic and organic acid addition salts of a compound. Pharmaceutically acceptable salts are those that do not cause severe irritation to the target substance and do not impair the biological activity and physical properties of the compound. Inorganic acid salts may be hydrochlorides, hydrobromic acids, phosphates, sulfates, or hydrogen sulfates. Organic acid salts may be formates, acetates, propionates, lactates, oxalates, tartrates, malates, maleates, citrates, fumarates, benzenesulfonates, camphorsulfonates, ethanedisulfonates, trichloroacetates, trifluoroacetates, benzoates, gluconates, methanesulfonates, glycolates, succinates, 4-toluenesulfonates, galacturonic acids, embolates, glutamates, ethanesulfonates, p-toluenesulfonates, or aspartate salts. Metal salts may be calcium, sodium, magnesium, strontium, or potassium salts.

[0130] Compounds of Formula 1 can be agonists of SOX9 (SRY-box transcription factor 9). SOX9 is known to recognize the CCTTGAG sequence along with other members of the HMG-box family of DNA-binding proteins and is expressed by proliferating chondrocytes rather than hypertrophic chondrocytes, which is crucial for progenitor cell differentiation into chondrocytes. Compounds of Formula 1 can increase SOX9 expression by inducing the aggregation of the SOX9 transcription factor. Compounds of Formula 1 can be SOX9 transcription factor aggregation inducers, SOX9 activators, or activators.

[0131] Another aspect of the present invention provides a pharmaceutical composition comprising the above-described compound, or a stereoisomer thereof, a solvate, or a pharmaceutically acceptable salt thereof.

[0132] Another aspect of the present invention provides a pharmaceutical composition for the prevention or treatment of diseases associated with reduced expression of SOX9 (SRY-box transcription factor 9), comprising the above-described compound, or a stereoisomer thereof, a solvate, or a pharmaceutically acceptable salt thereof.

[0133] The compounds, stereoisomers, solvates, pharmaceutically acceptable salts, and SOX9 are as described above.

[0134] Diseases associated with decreased SOX9 expression are attributed to SOX9 inhibition.

[0135] In some implementations, diseases associated with decreased SOX9 expression may be osteoarthritis and osteoarthritis-related diseases.

[0136] In some embodiments, the osteoarthritis-related disease may be chondropathy, osteonecrosis, or chronic pain.

[0137] The term "prevention" refers to any effect of inhibiting the onset of SOX9-related disease or delaying its progression by administering the pharmaceutical composition. The term "treatment" refers to any effect of improving or beneficially altering the symptoms of SOX9-related disease by administering the pharmaceutical composition.

[0138] The pharmaceutical composition may comprise a pharmaceutically acceptable carrier. The carrier is intended to encompass excipients, diluents, or adjuvants. The carrier may be selected from, for example, lactose, glucose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, polyvinylpyrrolidone, water, physiological saline, buffers such as PBS, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil. The composition may comprise fillers, anti-aggregating agents, lubricants, wetting agents, flavoring agents, emulsifiers, preservatives, or combinations thereof.

[0139] The pharmaceutical composition can be prepared into any formulation using conventional methods. The composition can be formulated, for example, into oral dosage forms (such as powders, tablets, capsules, syrups, pills, or granules) or parenteral dosage forms (such as injections). Furthermore, the composition can be prepared as a systemic or topical formulation.

[0140] In the pharmaceutical compositions, the solid dosage form for oral administration may be a tablet, pill, powder, granule, or capsule. The solid dosage form may further contain excipients. Examples of excipients include starch, calcium carbonate, sucrose, lactose, and gelatin. Furthermore, the solid dosage form may further contain lubricants such as magnesium stearate or talc. In the pharmaceutical compositions, the liquid dosage form for oral administration may be a suspension, oral liquid, emulsion, or syrup. The liquid dosage form may contain water or liquid paraffin. The liquid dosage form may contain excipients such as wetting agents, sweeteners, flavoring agents, or preservatives. In the pharmaceutical compositions, the dosage form for parenteral administration may be a sterile aqueous solution, a non-aqueous solution, a suspension, an emulsion, a lyophilized product, or a suppository. The non-aqueous solution or suspension may contain vegetable oils or esters. Examples of vegetable oils include propylene glycol, polyethylene glycol, and olive oil. Examples of esters are ethyl oleate. The base of suppositories can be vetexol, polyethylene glycol, Tween 61, cocoa butter, lauryl ester or glycerin gelatin.

[0141] The pharmaceutical composition comprises a compound, or a stereoisomer thereof, a solvate, or a pharmaceutically acceptable salt thereof, as the active ingredient of the pharmaceutical composition. "Active ingredient" refers to a physiologically active substance used to achieve pharmacological activity (e.g., treatment of diseases associated with reduced SOX9 expression).

[0142] The pharmaceutical composition may comprise an effective amount of a compound according to one aspect, or a stereoisomer thereof, solvate, or pharmaceutically acceptable salt thereof. The term "effective amount" refers to an amount sufficient to exert a preventive or therapeutic effect on a disease when administered to an individual requiring prevention or treatment. The effective amount may be appropriately selected by those skilled in the art based on the selected cells or individual. Preferred doses of the pharmaceutical composition will vary depending on the individual's condition and weight, the severity of the disease, the form of the drug, and the route and duration of administration, but may be appropriately selected by those skilled in the art. The effective amount may be about 0.5 μg to about 2 g per pharmaceutical composition, about 1 μg to about 1 g, about 10 μg to about 500 mg, about 100 μg to about 100 mg, or about 1 mg to about 50 mg. However, the compound, stereoisomer, solvate, or pharmaceutically acceptable salt may be administered, for example, in amounts from about 0.0001 mg / kg to about 100 mg / kg, or from about 0.001 mg / kg to about 100 mg / kg, divided into 1 to 24 daily doses, 1 to 7 doses every 2 days to 1 week, or 1 to 24 doses every 1 to 12 months. In the pharmaceutical composition, based on the total weight of the entire composition, the content of the compound, stereoisomer, solvate, or pharmaceutically acceptable salt may be from about 0.0001% by weight to about 10% by weight, or from about 0.001% by weight to about 1% by weight.

[0143] The composition can be administered orally or parenterally. Administration methods include, for example, oral, transdermal, subcutaneous, rectal, intravenous, intra-arterial, intraperitoneal, intramuscular, intrasternal, local, intranasal, intratracheal, or intradermal routes. The composition can be administered systemically or locally, and can be used alone or in combination with other pharmaceutically active compounds.

[0144] Another aspect of the present invention provides a method for preventing or treating diseases associated with reduced SOX9 expression, comprising administering to an individual the aforementioned compound, or a stereoisomer thereof, a solvate, or a pharmaceutically acceptable salt thereof.

[0145] The compounds, stereoisomers, solvates, pharmaceutically acceptable salts, SOX9, and diseases, prevention, and treatment associated with reduced SOX9 expression are as described above.

[0146] The individuals may be mammals, such as humans, mice, rats, cattle, horses, pigs, dogs, monkeys, sheep, goats, apes, or cats. The individuals may be those with disease symptoms associated with decreased SOX9 expression or those at high risk of developing the disease.

[0147] The method may further include administering to the individual an active ingredient known to be effective in preventing or treating SOX9-related diseases. According to one aspect, the known active ingredient may be administered simultaneously, separately, or sequentially with the aforementioned compound, stereoisomer, solvate, or pharmaceutically acceptable salt.

[0148] The drug can be administered orally or parenterally. Routes of administration include, for example, oral, transdermal, subcutaneous, rectal, intravenous, intra-arterial, intraperitoneal, intramuscular, intrasternal, local, intranasal, intratracheal, or intradermal routes. The drug composition can be administered systemically or locally, and can be used alone or in combination with other pharmaceutically active compounds.

[0149] The preferred dosage of the pharmaceutical composition varies depending on the patient's condition and weight, disease severity, drug form, route of administration, and duration of duration, and can be appropriately selected by those skilled in the art. Dosage may be, for example, about 0.001 mg / kg to about 100 mg / kg, about 0.01 mg / kg to about 10 mg / kg, or about 0.1 mg / kg to about 1 mg / kg for adults. Administration may be, for example, once daily, 2 to 24 times daily, 1 to 2 times every 3 days, 1 to 6 times weekly, 1 to 10 times every 2 weeks, 1 to 15 times every 3 weeks, 1 to 3 times every 4 weeks, or 1 to 12 times annually.

[0150] Another aspect of the invention provides a compound, or a stereoisomer thereof, solvate thereof, or a pharmaceutically acceptable salt thereof, for the prevention or treatment of diseases associated with reduced SOX9 expression.

[0151] The compounds, stereoisomers, solvates, pharmaceutically acceptable salts, SOX9, and diseases, prevention, and treatment associated with reduced SOX9 expression are as described above.

[0152] Another aspect of the invention provides the use of a compound, or a stereoisomer thereof, a solvate, or a pharmaceutically acceptable salt thereof, in the prevention or treatment of diseases associated with reduced SOX9 expression.

[0153] The compounds, stereoisomers, solvates, pharmaceutically acceptable salts, SOX9, and diseases, prevention, and treatment associated with reduced SOX9 expression are as described above.

[0154] Another aspect of the invention provides the use of the above-described compounds, or stereoisomers thereof, solvates, or pharmaceutically acceptable salts thereof, in the preparation of medicaments for the prevention or treatment of diseases associated with reduced SOX9 expression.

[0155] The compounds, stereoisomers, solvates, pharmaceutically acceptable salts, SOX9, and diseases, prevention, and treatment associated with reduced SOX9 expression are as described above.

[0156] Invention Effects

[0157] The compounds of the present invention, and pharmaceutical compositions comprising them for the prevention or treatment of diseases associated with reduced SOX9 expression, can effectively prevent or treat diseases associated with reduced SOX9 expression. Attached Figure Description

[0158] Figure 1 Data on the effects of the compounds in the examples on SOX9 transcription, evaluated by luciferase assays, are shown.

[0159] Figure 2a Histogram showing the distribution of transcriptional activity effects of compound SOX9 in the examples.

[0160] Figure 2b The box plot shows the transcriptional activity effect of whether the compounds in the examples induce SOX9 aggregation.

[0161] Figure 3a Fluorescence images of SOX9-overexpressing cell lysates after treatment with compounds B8 and C9.

[0162] Figure 3b This is a graph showing the effect of SOX9 transcriptional activity as measured using luciferase technology.

[0163] Figure 4a The fluorescence image shows the variation in the degree of SOX9 aggregate formation with the drug structure.

[0164] Figure 4bThis is a graph showing the changes in SOX9 transcriptional activity as a function of drug structure, as determined using luciferase technology.

[0165] Figure 5a This diagram illustrates the degree of disorder at different structures and locations of SOX9.

[0166] Figure 5b A graph illustrating the degree of SOX9 aggregation induced by different SOX9 deletion variants.

[0167] Figure 5c This diagram illustrates the degree of SOX9 aggregation induced by the drug on variants with deletions in the C-terminal disorder region of SOX9.

[0168] Figure 6 A graph illustrating the degree of SOX9 aggregation induced by drug-induced SOX9 C-terminal aromatic amino acid substitution variants.

[0169] Figure 7a This is a schematic diagram of the process for identifying proteins in drug-induced aggregates using SOX9-turboID.

[0170] Figure 7b The volcano plot shows that more protein was identified after drug treatment compared to the solvent used to dissolve the drug.

[0171] Figure 8a The histogram shows the degree of protein disorder frequently observed after drug treatment, compared to the solvent in which the drug was dissolved.

[0172] Figure 8b The histogram shows the pattern similarity of frequently observed proteins after drug treatment relative to the C-terminal aromatic amino acids of SOX9, compared to the solvent in which the drug was dissolved.

[0173] Figure 8c The histogram shows the relationship between the pattern similarity of frequently observed proteins after drug treatment relative to the SOX9 C-terminal aromatic amino acids and the conditional probability, compared to the solvent in which the drug was dissolved.

[0174] Figure 9 Images of SOX9 observed in the cell nucleus using conventional fluorescence microscopy and super-resolution microscopy (dSTORM).

[0175] Figure 10 The results are obtained by detecting the mRNA level of the SOX9 target gene using qPCR after drug treatment.

[0176] Figure 11 Fluorescence images of SOX9 target gene (Acan, Col9a1, Sox9) mRNA and SOX9 protein simultaneously detected by FISH.

[0177] Figure 12a The histogram shows the size distribution of SOX9 aggregates near the SOX9 target gene mRNA after drug treatment.

[0178] Figure 12b This is a schematic diagram showing the changes in the size of SOX9 aggregates near the SOX9 target gene mRNA after drug treatment.

[0179] Figure 13 This refers to the degree to which SOX9 binds to gene regulatory regions on DNA after drug treatment, compared to the solvent used to dissolve the drug.

[0180] Figure 14 This refers to the degree to which SOX9 binds to a single gene regulatory region on DNA after drug treatment, compared to the solvent used to dissolve the drug.

[0181] Figure 15a A schematic diagram of the experimental procedure for administering drugs to a mouse model of osteoarthritis.

[0182] Figure 15b Images showing the degree of cartilage tissue recovery in the joints of a mouse model of osteoarthritis after drug treatment (red: cartilage; blue: bone).

[0183] Figure 15c This is a graph showing the extent of tissue recovery after drug treatment, assessed using the OARSI score.

[0184] Figure 16 A schematic diagram and results of an experiment to evaluate the weight-bearing capacity of the legs of a mouse model of osteoarthritis after drug treatment relative to the untreated legs.

[0185] Figure 17 A schematic diagram and results of the experimental procedure for quantitatively detecting the tolerance of the soles of mice with osteoarthritis to acupuncture stimulation after drug treatment. Detailed Implementation

[0186] The invention will now be described in more detail by way of examples. However, these examples are intended to be illustrative, and the scope of the invention is not limited to these examples.

[0187] Preparation Example A1: Synthesis of 3-(2-aminothiazolyl-4-yl)-7-methoxycoumarin

[0188] Step 1: 3-Acetyl-7-methoxycoumarin

[0189] 2-Hydroxy-4-methoxybenzaldehyde (2 mmol, CAS: 673-22-3), ethyl acetoacetate (2.4 mmol), and piperidine (0.2 mmol) were stirred in ethanol (4 mL). After the reaction was complete, the reaction mixture was cooled in an ice-water bath, and the precipitate was filtered to give 3-acetyl-7-methoxycoumarin in 88% yield.

[0190] ¹H NMR (400 MHz, DMSO) δ 8.64 (s, 1H), 7.87 (d, J = 8.7 Hz, 1H), 7.06 (d, J = 2.3 Hz, 1H), 7.02 (dd, J = 8.7, 2.4 Hz, 1H), 3.90 (s, 3H), 2.56(s,3H).

[0191] Step 2: 3-Bromoacetyl-7-methoxycoumarin

[0192] The product from step 1 (1 mmol) was dissolved in dichloromethane (10 mL) with tetrabutylammonium tribromide (TBATB, 2 mmol) and stirred at room temperature. After the reaction was complete, the precipitate was separated by filtration to give 3-bromoacetyl-7-methoxycoumarin, with a yield of 59%.

[0193] ¹H NMR (400 MHz, DMSO) δ 8.82 (s, 1H), 7.93 (d, J = 8.7 Hz, 1H), 7.12(d, J = 2.4 Hz, 1H), 7.06 (dd, J = 8.7, 2.5 Hz, 1H), 4.87 (s, 2H), 3.92 (s,3H).

[0194] Step 3: 3-(2-aminothiazolyl-4-yl)-7-methoxycoumarin

[0195] The product from step 2 (1 mmol) was mixed with thiourea (2 mmol) in ethanol (10 mL) and stirred and heated. After the reaction was complete, the precipitate was filtered to give intermediate a-1 in 99% yield.

[0196] ¹H NMR (400 MHz, DMSO) δ 8.47 (s, 1H), 7.75 (d, J = 8.7 Hz, 1H), 7.42(s, 1H), 7.07 (d, J = 2.4 Hz, 1H), 7.00 (d, J = 8.6 Hz, 1H), 3.88 (s, 3H).

[0197] Preparation Examples A2 to A13

[0198] Except for replacing 2-hydroxy-4-methoxybenzaldehyde with the hydroxybenzaldehyde compound shown in Table 1 in step 1 of preparation example A1, the other operations were the same as in preparation example A1, and intermediates a-2 to a-12 were prepared respectively.

[0199] [Table 1]

[0200] Preparation Example A13: Synthesis of 3-(2-aminooxazol-4-yl)-7-methoxycoumarin

[0201] Intermediate a-13 (52%, 387 mg) was obtained in the same manner as in Preparation Example A1, except that urea (CAS: 57-13-6) was used instead of thiourea in step 3 of Preparation Example A1.

[0202] 1 H NMR (300 MHz, DMSO) δ 8.24 (s, 1H), 7.84 (s, 1H), 7.77 (d, J = 8.7Hz, 1H), 7.05 (d, J = 2.6 Hz, 1H), 6.98 (dd, J = 8.7, 2.4 Hz, 1H), 6.83 (s,2H), 3.87 (s,3H).

[0203] The compounds shown in Table 2 below are used as intermediates a-14 to a-17.

[0204] [Table 2]

[0205] Preparation Example A18: Synthesis of 4-(2-aminothiazol-4-yl)biphenyl

[0206] Intermediate a-18 (94%, 476 mg) was obtained in the same manner as in step 3 of preparation example A1, except that 2-bromo-4'-phenylacetophenone (CAS: 135-73-9) was used instead of the product in step 2 of preparation example A1.

[0207] ¹H NMR (400 MHz, DMSO) δ 8.85 (s, 2H), 7.88–7.78 (m, 4H), 7.78–7.69(m, 2H), 7.50 (dd, J = 8.5, 6.9 Hz, 2H), 7.44–7.36 (m, 1H), 7.32 (s, 1H).

[0208] Preparation Example A19: Synthesis of 3-(2-aminothiazol-4-yl)-6-methyl-2H-pyran-2,3(3H)-dione

[0209] Step 1: 3-(2-bromoacetyl)-6-methyl-2H-pyran-2,4(3H)-dione

[0210] Dehydroacetic acid (5.0 mmol, CAS: 520-45-6) and p-toluenesulfonic acid (5.5 mmol, CAS: 6192-52-5) were stirred in acetonitrile (10 mL). Then, NBS (5.5 mmol, CAS: 128-08-5) was added sequentially, and the mixture was heated. After 9 hours, water was added to the reaction mixture, the product was extracted with dichloromethane, and concentrated to give the final product (see Synthetic Communications, 2012, 42(18), 2739–2747).

[0211] Step 2: 3-(2-aminothiazolyl-4-yl)-6-methyl-2H-pyran-2,3(3H)-dione

[0212] Using the product from step 1, intermediate a-19 (8%, 94 mg) was obtained in the same manner as in step 3 of preparation example A1.

[0213] ¹H NMR (400 MHz, DMSO) δ 15.12 (s, 1H), 8.02 (s, 2H), 7.10 (s, 1H), 6.12 (d, J = 1.0 Hz, 1H), 2.20 (d, J = 0.9 Hz, 3H).

[0214] Preparation Example A20: Synthesis of (2-aminothiazolyl-5-yl)(phenyl)methyl ketone

[0215] Step 1: (E)-3-(dimethylamino)-1-phenylprop-2-en-1-one

[0216] Acetophenone (2.0 mmol) and N,N-dimethylformamide dimethyl acetal (4.0 mmol) were stirred and heated in toluene (2 mL) for 18 hours. After removing the solvent, (E)-3-(dimethylamino)-1-phenylprop-2-en-1-one (124 mg, 35%) was obtained by column chromatography (dichloromethane / methanol = 5:1).

[0217] Step 2: (2-aminothiazolyl-5-yl)(phenyl)methyl ketone

[0218] The product from step 1 (0.4 mmol), sulfur powder (1.7 mmol), cyanamide solution (1.7 mmol), and N-methylmorpholine (NMM, 0.1 mmol) were stirred in N-methylpyrrolidone (NMP, 1 mL). The mixture was then reacted at 100°C for 16 hours under an argon atmosphere. After the reaction was complete, the reaction mixture was cooled and diluted with ethyl acetate. Water was then added for extraction. The organic layer containing the product was dried over sodium sulfate to remove residual water, concentrated, and purified by column chromatography (n-hexane / ethyl acetate = 1:1) to give intermediate a-20 (31%, 78 mg) (see J. Org. Chem. 2019, 84(18), 12237–12245).

[0219] ¹H NMR (600 MHz, CDCl3) δ 7.78 (dd, J = 8.3, 1.4 Hz, 2H), 7.62 (s,1H), 7.60–7.56 (m, 1H), 7.53–7.46 (m, 2H), 6.22 (br s, 2H).

[0220] Preparation Example B1: Synthesis of 3-(2-succiniminothiazo-4-yl)-7-methoxycoumarin

[0221] Succinic anhydride (10 mmol, 1.00 g) and intermediate a-1 (1 mmol) were placed in a mortar, mixed thoroughly, and transferred to a reactor. The reaction mixture was stirred at 150°C for 5 hours. After the reaction was complete, the product was purified by silica gel chromatography (CH2Cl2: CH3C(O)CH3=20:1) to give intermediate b-1 (yield: 84%, 300 mg).

[0222] 1H NMR (400 MHz, DMSO) δ 8.65 (s, 1H), 8.39 (s, 1H), 7.84 (d, J = 8.7Hz, 1H), 7.09 (d, J = 2.4 Hz, 1H), 7.00 (dd, J = 8.7, 2.4 Hz, 1H), 3.88 (s,3H), 2.87 (s,4H).

[0223] Preparation Examples B2 to B21

[0224] Intermediate a-1 in Preparation Example B1 was replaced with intermediates a-2 to a-20 respectively to synthesize intermediates b-2 to b-20.

[0225] [Table 3]

[0226] Example 1: N 1 -(3,4-Dimethoxyphenethyl)-N 4 Synthesis of 4-(7-methoxycoumarin-3-yl)thiazolyl-2-yl)succinamide

[0227] In a reactor equipped with a condenser, intermediate b-1 (0.20 mmol) was dissolved in anhydrous dioxane (2 mL) under an argon atmosphere, and 2-(3,4-dimethoxyphenyl)ethyl-1-amine (0.22 mmol, CAS: 120-20-7, TCI / D0678) was added. The reaction mixture was stirred at 110°C for 5 hours. After the reaction was complete, the reaction mixture was concentrated under vacuum. Water (20 mL) was added to the resulting residue to precipitate the product. The precipitate was separated by centrifugation or filtration and then freeze-dried to give the compound of Example 1 (compound B8, yield: 89%, 96 mg).

[0228] 1H NMR (400 MHz, DMSO) δ 12.30 (s, 1H), 8.54 (s, 1H), 7.99 (t, J =5.6 Hz, 1H), 7.89 (s, 1H), 7.77 (d, J = 8.7 Hz, 1H), 7.09 (d, J = 2.4 Hz,1H), 7.00 (dd, J = 8.6, 2.5 Hz, 1H), 6.84 (d, J = 8.2 Hz, 1H), 6.80 (d, J =2.0 Hz, 1H), 6.70 (dd, J = 8.1, 2.0 Hz, 1H), 3.88 (s, 3H), 3.74 (s, 3H), 3.70 (s, 3H), 3.23 (dt, J = 6.7, 6.7 Hz, 2H), 2.68 (t, J = 6.9 Hz, 2H), 2.62 (t, J= 7.4 Hz, 2H), 2.44 (t, J = 7.0 Hz, 2H).

[0229] Examples 2 to 81

[0230] Except for the following modifications to intermediate b-1 and 2-(3,4-dimethoxyphenyl)ethyl-1-amine of Example 1, the compounds of Examples 2 to 81 were obtained in the same manner as in Example 1.

[0231] [Table 4]

[0232] [Table 5]

[0233] Experimental Example 1: Evaluation of SOX9 transcriptional activity

[0234] To confirm whether the compound could enhance SOX9 transcriptional activity, a luciferase assay, which can measure SOX9 transcriptional activity, was performed using the following experimental method.

[0235] Primary chondrocytes were extracted from mouse cartilage. The extracted cells were stably cultured in a hypoxic incubator (O2 3%, CO2 5%). The culture medium used was DMEM (Dulbecco modified Eagle medium, LM001-05, Welgene) supplemented with 10% fetal bovine serum (FBS, Gibco).

[0236] After the cells stabilized, they were treated with Sigma-Aldrich hyaluronidase (IS type) for 4 hours for luciferase plasmid transfection.

[0237] During transfection, Metafectene Pro transfection reagent (Biontex) was used to co-treat cells for 6 hours with a 4X48pCol2a1-luciferase plasmid expressing SOX9 transcriptionally promoted activity and a constitutive Renilla luciferase plasmid as a transfection control. The 4X48pCol2a1-luciferase plasmid and Renilla luciferase plasmid were described as described in Kim, S. et al., “Tankyrase inhibition preserves osteoarthritic cartilage by coordinating cartilage matrix anabolism via effects on SOX9 PARylation,” Nat Commun 10, 4898 (2019).

[0238] Six hours later, the culture medium was replaced with one containing the compounds. Here, each compound was dissolved in DMSO at a concentration of about 5 mM to 10 mM and then added to the culture medium to bring the final concentration to 7.5 μM.

[0239] Forty-eight hours later, cells were lysed, and the luciferase substrate was reacted using a Dual Luciferase Assay Kit (Promega). The luminescence value of the luciferase (relative to DMSO) was measured using a microplate reader. Results were graded according to the following criteria and are listed in Table 6. Figure 1 The graph. 1.

[0240] ++++: 8 or higher / +++: less than 8, 5 or higher / ++: less than 5, 2 or higher / +: less than 2, higher than 1

[0241] [Table 6]

[0242] Refer to Table 6 and Figure 1 This confirms that the compound of the present invention has excellent SOX9 transcriptional activity.

[0243] The compounds in each example exhibited varying degrees of transcriptional activity enhancement effects. Figure 2a When examining changes in transcriptional activity induced by aggregation, a statistically significant correlation was found between aggregation and the effect on transcriptional activity. Figure 2b This means that, at the SOX9 protein level, the aggregation-inducing ability of a drug affects and is related to its efficacy.

[0244] Experimental Example 2: Induction of SOX9 condensation and evaluation of transcriptional activity

[0245] In a 100-mm culture dish, inoculate 9 × 10⁹ cells / mL. 5 Cells were selected for transfection and grown in a cell culture incubator for two days. Using a transfection reagent (Poly-jet), 7 μg of the SOX9-eGFP plasmid, which is capable of intracellular overexpression, was used to transfect the growing cells. Cells were then exposed to FBS-free DMEM medium for 30 minutes. After 30 minutes, the SOX9-eGFP plasmid and Polyjet were mixed in DMEM, and cells were added after a 10-minute interval. After reacting in the incubator for 4 hours, FBS-containing DMEM medium was added, and the cells were incubated for two days. Subsequently, the cells were separated using a scraper and centrifuged. The cell pellet was frozen in liquid nitrogen and stored at -80°C.

[0246] Cell lysis buffer containing 0.2% Triton X-100, 1 mM EDTA, 50 mM HEPES, 150 mM NaCl, 10% glycerol, 1% protease inhibitor mixture, 1% phosphatase inhibitor mixture (tyrosine, serine / threonine), and 2 mM TCEP was added to the cell pellet to disperse the pellet. The dispersed cells were lysed using an ultrasonic homogenizer, and the lysate was centrifuged (15,000 × g, 10 min) to obtain the supernatant. The total protein concentration of the obtained supernatant was measured using a DC (detergent-compatible) assay, and the SOX9-eGFP concentration was measured using a fluorometer. Cell lysates after concentration measurement were used immediately or frozen in liquid nitrogen and stored at -80°C.

[0247] SOX9, which is relatively difficult to purify in E. coli, was expressed and purified from HEK293T using the Halo-Tag purification method. Even when using SOX9 (instead of lysis buffer), the correlation between aggregation induction and transcriptional activity was reproduced, thus confirming the effectiveness of using lysis buffer. Figure 2b ).

[0248] The compound from the examples was diluted to 1 / 100 of the desired concentration, and then transferred to tubes according to the reaction volume. The prepared cell lysate was added to a tube containing compound B8 or compound C9 and reacted. The lysate after reaction with the drug was transferred to a 96-well plate with a coverslip surface and reacted at room temperature for 1 hour until the reaction was complete. The 96-well plate after the reaction was completed was transferred to a confocal fluorescence microscope for fluorescence imaging.

[0249] The captured fluorescence images were analyzed using internal MATLAB code. K-means clustering was used to distinguish between aggregates and non-aggregates, and the fluorescence intensity, area, shape, and quantity of aggregates were analyzed.

[0250] The results confirmed that, compared with DMSO, compounds B8 and C9 induced the aggregation of SOX9. Figure 3a Meanwhile, using luciferase plasmids that specifically respond to SOX9 transcriptional activity, it was confirmed that compounds B8 and C9, which induce aggregation, increase the activity of the SOX9 transcription factor. Figure 3b In particular, compound B8 increased SOX9 transcriptional activity by approximately 2-fold, while compound C9 significantly enhanced activity by approximately 8-fold. Here, due to structural changes in compound C9, SOX9 condensation was lost upon individual atom changes (such as changes in the position or substitution of the nitrogen atom), and it was confirmed whether this condensation was induced to influence the regulation of transcriptional activity. Figure 4a and 4b This indicates that the induction of aggregation and the regulation of transcriptional activity occur based on the chemical structure of the compounds.

[0251] Experimental Example 3: Evaluation of the degree of aggregation in SOX9-deficient regions

[0252] The mechanism by which compound C9 helps enhance cartilage regeneration was investigated.

[0253] Specifically, the drug binding site was investigated based on C9, the most effective compound identified so far. SOX9 was divided into structured and unstructured regions; in particular, the IDR (Index Degree Ratio) of the unstructured region was very large, constituting approximately 80% of the protein. Figure 5a To investigate which interaction region induced the aggregation effect of compound C9, SOX9 variants with each region deleted were prepared. Cell lysates of the SOX9 variants were prepared as in the experimental example above, and it was confirmed that the aggregation phenomenon disappeared when C9 in the IDR region corresponding to the C-terminus of the protein was deleted. Figure 5b Furthermore, to reduce the binding site, deletion variants were constructed across the C-terminal region at 35-amino acid intervals, and cell lysates of these variants were prepared and examined. Here, when regions corresponding to the aggregation of aromatic amino acids 8 and 9 were deleted, the aggregation phenomenon was confirmed to be drastically reduced. Figure 5c ).

[0254] Experimental Example 4: Changes in SOX9 aggregation degree based on aromatic amino acid mutations in C-terminal regions 8 and 9 of SOX9

[0255] Cell lysates expressing mutants with alanine substitutions for aromatic amino acids were prepared, and the importance of aromatic amino acids was confirmed. Similar to variants where regions 8 and 9 were deleted, mutations in the aromatic amino acids corresponding to regions 8 and 9 reduced aggregation induced by compound C9. Figure 6This confirms that compound C9 induces SOX9 aggregation by interacting with the region of aromatic amino acid aggregation in the unstructured region of the SOX9 C-terminus.

[0256] Experimental Example 5: Protein Identification in Aggregation-Induced SOX9 Aggregates

[0257] Since aggregates are typically formed from multiple proteins rather than a single protein, to identify proteins that co-aggregated with SOX9 after treatment with compound C9, SOX9 was fused to TurboID, and proteins located within 10 nm of SOX9 were biotinylated after treatment with compound C9. Biotin-bound proteins were separated using streptavidin beads and analyzed by liquid chromatography-mass spectrometry (LC / MS). Figure 7a Each mass spectrometry experiment was performed three times for each condition. Mass spectrometry analysis revealed that proteins observed statistically significantly more frequently than those in the C9 treatment group compared to DMSO were identified as proteins involved in transcriptional regulation, such as Pol2, YTHDF3, CCAR2, MBNL1, NDUFA10, MYO1C, TAF5, and chromatin remodeling factors (KAT6A, KAT8). Proteins known to co-act with SOX9, such as SOX5 and SOX6, were also detected. Figure 7b ).

[0258] Experiment Example 6: Confirming protein disorder and evaluating similarity to SOX9-C-IDR through spatial frequency analysis.

[0259] Metapredict was used to identify the IDR regions of proteins significantly detected in C9 and DMSO-treated samples using sequence information. Here, it was confirmed that the proteins detected after C9 treatment contained a significant amount of IDR (indicating a higher concentration of IDR regions compared to those treated with DMSO). Figure 8a Since IDR is known to play an important role in condensation, this indirectly indicates that compound C9 induces SOX9 condensation.

[0260] Because compound C9 induces aggregation by targeting aromatic amino acids in the SOX9 IDR, mass spectrometry analysis was used to confirm whether the detected protein's IDR contained the target of compound C9. A one-dimensional binary vector was constructed, where aromatic amino acids in the protein's IDR were assigned a value of 1, and other amino acids were assigned a value of 0. Wavelet transform was then applied to it. The transformed information contained aromatic amino acid density information as a function of time (position) and frequency. Cross-correlation analysis was used to compare the similarity with the SOX9 C-IDR using the wavelet transform data of the detected protein. It was found that in samples treated with compound C9, IDRs with a similar pattern to the SOX9 C-IDR had a higher probability of having a higher similarity score. Figure 8b Since proteins with high similarity to the SOX9 C-IDR pattern are inherently rare, to rule out the possibility that this is the reason for their low detection frequency, conditional probabilities were calculated by considering the number of proteins corresponding to each similarity score. This revealed that IDRs with high similarity scores were more likely to be detected in samples treated with compound C9. Figure 8c These results support the fact that compound C9 can specifically induce aggregate formation based on a specific pattern (spatial frequency) of aromatic amino acid production in SOX9 C-IDR.

[0261] Experimental Example 7: Analysis of SOX9 aggregates using super-resolution fluorescence microscopy dSTORM

[0262] To investigate the effect of drugs on changes in intracellular SOX9 aggregates, SOX9 was labeled using IF (immunofluorescence staining) and fluorescence imaging was performed. Because transcription factor aggregates such as SOX9 aggregates have a size of approximately 100 nm to 1 μm, changes in SOX9 aggregates cannot be detected at the level of conventional fluorescence microscopy. Therefore, dSTORM (direct random optical reconstruction microscopy, a type of super-resolution microscopy) was used to obtain... Figure 9 Fluorescence images.

[0263] Experiment Example 8: Evaluation of mRNA levels and RNA-FISH analysis of genes involved in cartilage regeneration

[0264] To investigate the relationship between observed changes in SOX9 aggregates and transcriptional activity, RNA FISH (fluorescence in situ hybridization) and dSTORM were performed on Col9a1, Acan, and SOX9 (transcriptants with highly upregulated mRNA levels in C9-treated samples compared to DMSO), providing... Figure 10 mRNA level map and Figure 11 dSTORM fluorescence images.

[0265] RNA-FISH confirmed that the locations of RNA foci observed in the cell nucleus co-localized well with the locations of SOX9 aggregates. Figure 11 Image analysis revealed that the size of SOX9 aggregates observed near RNA foci in C9, compared to DMSO, increased by approximately 30% to 50%. Figure 12a and 12b This is a high level compared to the 2% size change of SOX9 condensates in the cell nucleus. Therefore, compound C9 was found to be involved in altering the size of SOX9 condensates near RNA, particularly regarding transcriptional activity.

[0266] Experiment Example 9: Confirming the effect of compounds on the degree of SOX9-DNA binding using Cut & Tag

[0267] To investigate whether drug-modified SOX9 aggregates actually participate in transcriptional activity, a cut & tag experiment was performed to identify genome-wide DNA regions bound by SOX9. The study confirmed that drug treatment increased SOX9 binding to DNA. Figure 13 ).

[0268] Experiment 10: Confirming the changes in the degree of binding between SOX9 and the DNA of genes involved in cartilage regeneration after compound treatment.

[0269] When information on SOX9 binding to DNA obtained via Cut & Tag was verified against SOX9, Col2a1, Col9a1, Chad, Acan, and Comp (genes involved in cartilage formation), treatment with compound C9 was found to increase SOX9 binding across the entire DNA (SOX9, Col2a1, and Chad), or to increase binding in specific regions of the DNA (red boxes) (Col9a1, Acan, and Comp) (DMSO data subtracted from C9 data, shown in yellow). Figure 14 These results indicate that compound C9 affects the size variation and DNA binding degree of SOX9 aggregates.

[0270] Experimental Example 11: Evaluation of the degree of cartilage tissue recovery in a mouse model of osteoarthritis

[0271] To confirm whether compound C9 promotes cartilage formation, experiments were conducted in a mouse model of osteoarthritis (OA) induced by DMM (medial meniscus instability) surgery, in which compound C9 was injected intra-articularly seven times at one-week intervals to achieve an intra-articular concentration of 750 μM. Figure 15aIn mouse tissues treated with DMSO, recovery did not occur and damage was observed (yellow triangles). In contrast, when treated with the drug, the damaged cartilage tissue was found to have recovered. Figure 15b To objectively evaluate cartilage tissue recovery, OARSI grades were assessed by individual researchers using a blinded test (higher grades indicate greater damage to cartilage tissue). It was confirmed that treatment with compound C9 resulted in statistically significantly lower OARSI grades. Figure 15c This indicates that compound C9 can help restore damaged cartilage tissue.

[0272] Experiment Example 12: Evaluating the degree of osteoarthritis relief through behavioral experiments

[0273] To confirm recovery from osteoarthritis at the behavioral level, weight-bearing and Von-Frey behavioral tests were performed. The weight-bearing test is an experiment measuring weight-bearing capacity in the operated or non-operated leg after drug treatment to confirm changes in weight-bearing capacity following treatment with compound C9. Figure 16 When compound C9 was administered to DMM-induced osteoarthritis (OA) mice, the weight-bearing level was comparable to that observed in uninduced OA mice, and the degree of change was significant relative to the DMSO-treated group. Figure 16 ).

[0274] The Von-Frey behavioral experiment assesses cartilage recovery in mice by gently stimulating the sole of their paws and recording the intensity of the stimulation as the mice retract their paws. Figure 17 When cartilage recovery improved, the mice were able to tolerate increased stimulus intensity. Compared to mice that had not undergone OA induction, OA-induced mice were found to tolerate greater stimulus intensity after treatment with compound C9. Figure 17 ).

[0275] These behavioral experimental results confirmed that compound C9 alleviates osteoarthritis at the behavioral level. Therefore, this invention has validated the ability to screen compounds like compound C9, which are cartilage regeneration substances capable of inducing SOX9 aggregation and thereby exhibiting cartilage regeneration effects.

Claims

1. A compound represented by Formula 1, or a stereoisomer, solvate, or pharmaceutically acceptable salt thereof: [Formula 1] in, In Equation 1, Y 1 and Y 2 One of them is N, and the other is S, O, or NR. a ¹; Y 1 and Y 2 Each can be independently classified as O, S, or NR. a ²; U is NR n2 Five- to seven-membered heterocyclic groups containing one to three nitrogen atoms, or structural segments formed by the interconnection of these groups; Z 1 C 1-6 Alkylene; Z 2 For direct keys, C 1-6 Alkylene, -NR n3 CO-, or a 5- to 7-membered heteroaryl group containing 1 to 3 heteroatoms selected from N, O and S; R a1 and R a2 Each can be H or C independently. 1-6 alkyl; R n1 R n2 and R n3 Each independently is H or C 1-6 alkyl; When Z 2 C 1-6 In the case of alkylene groups, any carbon atom of the alkylene group may optionally be converted by a halogen, a hydroxyl group, or a C-type carbon atom. 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 Alkyl, carboxyl, C 1-6 alkoxycarbonyl, carbamoyl, C 1-6 Alkyl carbamoyl, di(C 1-6 Alkyl) carbamoyl, cyano, nitro, oxo, or C 6-12 Aryl substitution; Z 3 For direct keys or -C(=O)-; Ring A is a 6- to 14-membered aryl group, a partially unsaturated 9- to 14-membered bicyclic carbocyclic group, a 5- to 12-membered heteroaryl group containing 1 to 3 heteroatoms selected from O, N or S, a 5- to 8-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N or S, or a fused benzo[a]heterocyclic group formed by fusion of a benzene ring with a 5- to 7-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N or S. Ring A can be arbitrarily divided by 1 to 3 Rs A Replace; and R A Choose free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 The group consisting of alkyl, cyano, nitro, and oxo groups; The ring E is a 6- to 14-membered aryl group, a partially unsaturated 9- to 14-membered bicyclic carbocyclic group, a 5- to 12-membered heteroaryl group containing 1 to 3 heteroatoms selected from O, N or S, a 5- to 12-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N or S, or a fused heterocyclic aryl group formed by fusion of a 5- to 7-membered heterocycle containing 1 to 3 heteroatoms selected from O, N or S with a phenyl group; Ring E can be arbitrarily divided by 1 to 3 R E Replace; and R E Choose free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, carboxyl, C 1-6 alkoxycarbonyl, carbamoyl, C 1-6 Alkyl carbamoyl, di(C 1-6 Alkyl) carbamoyl, halosulfonyl, sulfinyl, C 1-6 alkylsulfonyl, cyano, nitro, oxo, or optionally substituted with one or more groups selected from halogen, hydroxyl, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Substituents of alkyl, amino, nitro and cyano groups on C 1-6 A group composed of alkyl groups.

2. The compound according to claim 1, or its stereoisomers, solvates, or pharmaceutically acceptable salts, The compound mentioned above is selected from compounds shown in Formula 2 or Formula 3: [Equation 2] [Formula 3] in, In equations 2 and 3, Z 1 Z 2 X 1 X 2 ,U,R n1 R n2 Ring A and ring E are as defined in claim 1.

3. The compound according to claim 2, or its stereoisomers, solvates, or pharmaceutically acceptable salts, In equation 2, Y 1 Y in formula 3 2 Each can be either S or O independently.

4. The compound according to claim 1, or its stereoisomers, solvates, or pharmaceutically acceptable salts, U is selected from NR n2 Piperidine dimethyl, piperazine dimethyl, or NR n2 Combinations with piperidinediyl or piperazinediyl.

5. The compound according to claim 1, or its stereoisomers, solvates, or pharmaceutically acceptable salts, Among them, U is free to choose NR n2 The group consisting of the following structures, wherein 1 Connect to -C(=X) 2 carbon atoms of )-, and 2 Connect to Z 2 : 。 6. The compound according to claim 1, or its stereoisomers, solvates, or pharmaceutically acceptable salts, Wherein ring A is phenyl, biphenyl, tetrahydronaphthyl, pyranyl, pyranone, benzopyranyl, or benzopyranone.

7. The compound according to claim 1, or its stereoisomers, solvates, or pharmaceutically acceptable salts, Where ring A is or Furthermore, ring A can be arbitrarily divided by 1 to 3 R's. A replace.

8. The compound according to claim 7, or its stereoisomers, solvates, or pharmaceutically acceptable salts thereof. Z 3 It is a direct bond, and the ring A is , ,or ;or Z 3 It is -C(=O)-, and ring A is or .

9. The compound according to claim 1, or its stereoisomers, solvates, or pharmaceutically acceptable salts thereof. Where R A Choose from the group consisting of F, Cl, Br, hydroxyl, methoxy, dimethylamino, methyl, and oxo.

10. The compound according to claim 1, or its stereoisomer, solvate, or pharmaceutically acceptable salt. Ring A is selected from the following chemical structures: 。 11. The compound according to claim 1, or its stereoisomers, solvates, or pharmaceutically acceptable salts, The ring E can be phenyl, naphthyl, indanyl, pyridinyl, pyridinyl, imidazolyl, pyrazinyl, imidazolyl, pyrazolyl, triazolyl, thiophene, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, morpholinyl, thiomorpholinyl, piperidinyl, hexahydroxypyridinyl, hexahydroxypyrimidinyl, piperazinyl, indolyl, inazolyl, benzimidazolyl, quinolinyl, isoquinolinyl, cenolinyl, quinazolinyl, quinoxolinyl, phthalazinyl, or benzodioxolane.

12. The compound according to claim 1, or its stereoisomers, solvates, or pharmaceutically acceptable salts, The ring E is selected from the chemical structures shown below, and the ring E is optionally separated by 1 to 3 R groups. E replace: 。 13. The compound according to claim 1, or its stereoisomers, solvates, or pharmaceutically acceptable salts, Where R E The group consisting of methoxy, hydroxy, amino, F, Cl, Br, trifluoromethyl, cyano, hydroxymethyl, methyl, fluorosulfonyl, nitro, and oxo is selected.

14. The compound according to claim 1, or its stereoisomers, solvates, or pharmaceutically acceptable salts thereof. Ring E is selected from the following chemical structures: 。 15. The compound according to claim 1, or its stereoisomers, solvates, or pharmaceutically acceptable salts thereof. The compound is represented by formula I, formula II or formula III: [Formula I] in, In equation I, Y 1 For S, O or NR a1 ; R a1 For H or C 1-6 alkyl; R n1 and R n2 Each independently is H or C 1-6 alkyl; Z 4 For direct key or C 1-4 Alkylene; R 1 H, halogen, hydroxyl, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 Alkyl, carboxyl, C 1-6 alkoxycarbonyl, carbamoyl, C 1-6 Alkyl carbamoyl, di(C 1-6 Alkyl) carbamoyl, cyano, nitro, oxo or C 6-12 Aryl; Ring A is a 6- to 14-membered aryl group, a partially unsaturated 9- to 14-membered bicyclic carbocyclic group, a 6- to 12-membered heteroaryl group containing 1 to 3 heteroatoms selected from O, N or S, a 5- to 8-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N or S, or a fused benzo[a]heterocyclic group formed by fusion of a benzene ring with a 5- to 7-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N or S; Ring A can be arbitrarily divided by 1 to 3 Rs A Replace; and R A Choose free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 The group consisting of alkyl, cyano, nitro, and oxo groups; The ring E is a 6- to 14-membered aryl group, a partially unsaturated 9- to 14-membered bicyclic carbocyclic group, a 5- to 12-membered heteroaryl group containing 1 to 3 heteroatoms selected from O, N or S, a 5- to 12-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N or S, or a fused heterocyclic aryl group formed by fusion of a 5- to 7-membered heterocycle containing 1 to 3 heteroatoms selected from O, N or S with a phenyl group; Ring E can be arbitrarily divided by 1 to 3 R E Replace; and R E Choose free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, carboxyl, C 1-6 alkoxycarbonyl, carbamoyl, C 1-6 Alkyl carbamoyl, di(C 1-6 Alkyl) carbamoyl, halosulfonyl, sulfinyl, C 1-6 alkylsulfonyl, cyano, nitro, oxo, or optionally substituted with one or more groups selected from halogen, hydroxyl, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Substituents of alkyl, amino, nitro and cyano groups on C 1-6 The group consisting of alkyl groups; [Formula II] In Equation II, Y 2 For S, O or NR a1 ; R a1 For H or C 1-6 alkyl; R n1 and R n2 Each independently is H or C 1-6 alkyl; Z 4 For direct key or C 1-4 Alkylene; R 1 H, halogen, hydroxyl, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 Alkyl, carboxyl, C 1-6 alkoxycarbonyl, carbamoyl, C 1-6 Alkyl carbamoyl, di(C 1-6 Alkyl) carbamoyl, cyano, nitro, oxo or C 6-12 Aryl; Ring A 1 It is a 6- to 14-membered aryl group or a partially unsaturated 9- to 14-membered bicyclic carbocyclic group; Ring A 1 Optionally by 1 to 3 R A Replace; and R A Choose free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 The group consisting of alkyl, cyano, nitro, and oxo groups; The ring E is a 6- to 14-membered aryl group, a 5- to 12-membered heteroaryl group containing 1 to 3 heteroatoms selected from O, N or S, a 5- to 12-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N or S, or a fused heterocyclic aryl group formed by fusion of a 5- to 7-membered heterocycle containing 1 to 3 heteroatoms selected from O, N or S with a phenyl group; Ring E can be arbitrarily divided by 1 to 3 R E Replace; and R E Choose free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 Alkyl, carboxyl, C 1-6 alkoxycarbonyl, carbamoyl, C 1-6 Alkyl carbamoyl, di(C 1-6 Alkyl) carbamoyl, halosulfonyl, sulfinyl, C 1-6 The group consisting of alkylsulfonyl, cyano, nitro and oxo groups; [Formula III] In Equation III, Z 1 For direct bond or NR n2 ; Y 3 and Y 4 At least one of them is N, and the other is CH; Z 5 For direct keys, C 1-6 Alkylene, -NR n3 CO-, or a 5- to 7-membered heteroaryl group containing 1 to 3 heteroatoms selected from N, O and S; R n1 R n2 and R n3 Each is independently an H or C1-6 alkyl group; Ring A is a 6- to 14-membered aryl group, a partially unsaturated 9- to 14-membered bicyclic carbocyclic group, a 5- to 12-membered heteroaryl group containing 1 to 3 heteroatoms selected from O, N or S, a 5- to 8-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N or S, or a fused benzo[a]heterocyclic group formed by fusion of a benzene ring with a 5- to 7-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N or S. Ring A can be arbitrarily divided by 1 to 3 Rs A Replace; and R A Choose free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 The group consisting of alkyl, cyano, nitro, and oxo groups; The ring E is a 6- to 14-membered aryl group, a partially unsaturated 9- to 14-membered bicyclic carbocyclic group, a 5- to 12-membered heteroaryl group containing 1 to 3 heteroatoms selected from O, N or S, a 5- to 12-membered heterocyclic group containing 1 to 3 heteroatoms selected from O, N or S, or a fused heterocyclic aryl group formed by fusion of a 5- to 7-membered heterocycle containing 1 to 3 heteroatoms selected from O, N or S with a phenyl group; Ring E can be arbitrarily divided by 1 to 3 R E Replace; and R E Choose free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, carboxyl, C 1-6 alkoxycarbonyl, carbamoyl, C 1-6 Alkyl carbamoyl, di(C 1-6 Alkyl) carbamoyl, halosulfonyl, sulfinyl, C 1-6 alkylsulfonyl, cyano, nitro, oxo, or optionally substituted with one or more groups selected from halogen, hydroxyl, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Substituents of alkyl, amino, nitro and cyano groups on C 1-6 The group consisting of alkyl groups; and s is 0 or 1.

16. The compound according to claim 15, or a stereoisomer, solvate, or pharmaceutically acceptable salt thereof. The compounds described herein are represented by formula IA, IB, IC, ID, IE, or IIA: [Form IA] [Form IB] [Form IC] [Formula ID] [Formula IE] [Form IIA] in, In formulas IA, IB, IC, ID, IE, or IIA R A Choose free halogens, hydroxyl groups, C 1-6 Alkoxy, amino, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, C 1-6 The group consisting of alkyl, cyano, nitro, and oxo groups; n, m1, m2, o1, o2, p, and q are each independent integers from 0 to 3; The sum of m1 and m2, and the sum of o1 and o2, are each 3 or less; and Y 1 Y 2 R n1 R n2 Z 4 R 1 And ring E as defined in claim 13.

17. The compound according to claim 1, or its stereoisomers, solvates, or pharmaceutically acceptable salts thereof. The compounds mentioned above are selected from compounds with the following chemical structures: 。 18. A pharmaceutical composition for the prevention or treatment of diseases associated with reduced expression of SOX9 (SRY box transcription factor 9), said pharmaceutical composition comprising a compound according to any one of claims 1 to 17, or a stereoisomer, solvate, or pharmaceutically acceptable salt thereof.

19. The pharmaceutical composition according to claim 18, The disease associated with decreased SOX9 expression is osteoarthritis.

20. A method for treating a disease associated with reduced SOX9 expression, the method comprising administering to an individual a compound according to any one of claims 1 to 17, or a stereoisomer, solvate, or pharmaceutically acceptable salt thereof.

21. Use of the compound according to any one of claims 1 to 17, or a solvate, stereoisomer, or pharmaceutically acceptable salt thereof, in the prevention or treatment of diseases associated with reduced SOX9 expression.

22. Use of the compound of any one of claims 1 to 17, or a solvate, stereoisomer, or pharmaceutically acceptable salt thereof, in the preparation of a medicament for the prevention or treatment of diseases associated with reduced SOX9 expression.

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