KR20230155324A - Derivatives of Piperidinedione - Google Patents

Abstract

A novel piperidinedione derivative compound is disclosed.

Description

Piperidinedione derivatives {Derivatives of Piperidinedione}

The present invention relates to piperidinedione derivative compounds. More specifically, the present invention relates to a piperidinedione derivative compound having a structure in which a target protein ligand and an E3 enzyme ligand are linked.

PROTACs, which stands for Proteolysis Targeting Chimeras, is a technology that induces the degradation of target proteins (POI: protein of interest). Instead of the term Protak, the term TPD (Targeted Protein Degradation) is sometimes used.

ProTac is a structure in which a target protein ligand and an E3 enzyme ligand are connected by a linker (see Figure 1). A target protein ligand is a structure that can bind to a target protein related to a disease, and an E3 enzyme ligand is a structure that can bind to an E3 enzyme.

E3 enzymes (E3 ligases) can label target proteins with ubiquitin, and these ubiquitin-labeled target proteins are degraded by the proteasome.

If the target protein ligand of the Protac compound binds to the target protein, the E3 enzyme is located very close to the target protein by the Protak compound, thereby creating an environment in which the target protein can be removed by the proteasome. do.

In particular, if the Protac compound binds to a target protein involved in the proliferation of cancer cells, it can induce the decomposition of the target protein, resulting in a cancer treatment effect.

WO 2010/078897 A1;

Taavi K. Neklesa et al., “Targeted protein degradation by PROTACs”; Pharmacology & Therapeutics 174 (2017) 138-144.

The present invention is intended to provide a compound that can be used for TPD by increasing the decomposition effect of target proteins (particularly c-MET protein).

The present invention relates to a compound of formula (1) or a pharmaceutically acceptable salt thereof:

[Formula 1]

In the above formula,

Z is the targeting protein ligand,

Q ₁ to Q ₄ are each independently CZ, CH, C-Hal or C≡N (where Hal is halogen), provided that any one of Q ₁ to Q ₄ is CZ,

R ¹ is hydrogen, halogen, nitro, amino, straight-chain, branched-chain or cyclic alkyl of C ₁ to C ₆ , or straight-chain, branched-chain or cyclic alkyl of C ₁ to C ₆ substituted by halogen.

In the compound of the present invention, R ¹ is preferably hydrogen, halogen, straight-chain, branched-chain or cyclic alkyl of C ₁ to C ₄ , or straight-chain, branched-chain or cyclic alkyl of C ₁ to C ₄ substituted with halogen. do.

In the compound of the present invention, a compound that is a c_MET inhibitor may be used as the target protein ligand, and preferably a compound selected from the group consisting of the following compounds may be used:

Additionally, the present invention relates to a compound of the following formula (2) or a pharmaceutically acceptable salt thereof:

[Formula 2]

In the above formula,

Y is -C=O or -CH ₂ ,

Z is the targeting protein ligand,

Q ₁ to Q ₄ are each independently CZ, CH, C-Hal or C≡N (where Hal is halogen), provided that any one of Q ₁ to Q ₄ is CZ.

In the compound of the present invention, a compound that is a c-MET inhibitor may be used as the target protein ligand, and preferably a compound selected from the group consisting of the following compounds may be used:

The compound according to the present invention may preferably be a compound selected from the group consisting of the following compounds or a pharmaceutically acceptable salt thereof:

Additionally, the compound according to the present invention may preferably be a compound selected from the group consisting of the following compounds or a pharmaceutically acceptable salt thereof:

3-(6-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]tria Zolo[4,5-b]pyrazin-1-yl)methyl)morpholino)pyrimidin-5-yl)benzyl)piperazin-1-yl)-1-oxophthalazin-2(1H)-yl ) piperidine-2,6-dione

3-(4-methyl-6-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2 ,3] triazolo [4,5-b] pyrazin-1-yl) methyl) morpholino) pyrimidin-5-yl) benzyl) piperazin-1-yl) -1-oxophthalazine-2 ( 1H)-yl)piperidine-2,6-dione

3-(7-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]tria Zolo[4,5-b]pyrazin-1-yl)methyl)morpholino)pyrimidin-5-yl)benzyl)piperazin-1-yl)-1-oxophthalazin-2(1H)-yl ) piperidine-2,6-dione

3-(8-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]tria Zolo[4,5-b]pyrazin-1-yl)methyl)morpholino)pyrimidin-5-yl)benzyl)piperazin-1-yl)-1-oxophthalazin-2(1H)-yl ) piperidine-2,6-dione

3-(4-methyl-8-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2 ,3] triazolo [4,5-b] pyrazin-1-yl) methyl) morpholino) pyrimidin-5-yl) benzyl) piperazin-1-yl) -1-oxophthalazine-2 ( 1H)-yl)piperidine-2,6-dione

3-(6-fluoro-7-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1, 2,3]triazolo[4,5-b]pyrazin-1-yl)methyl)morpholino)pyrimidin-5-yl)benzyl)piperazin-1-yl)-1-oxophthalazine-2 (1H)-yl)piperidine-2,6-dione

2-(2,6-dioxopiperidin-3-yl)-4-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazole-4 -yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl)methyl)morpholino)pyrimidin-5-yl)benzyl)piperazin-1-yl) Isoindoline-1,3-dione

2-(2,6-dioxopiperidin-3-yl)-5-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazole-4 -yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl)methyl)morpholino)pyrimidin-5-yl)benzyl)piperazin-1-yl) Isoindoline-1,3-dione

3-(1-(3-(5-((1-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl )piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

3-(1-(3-(5-((1-(2-(2,6-dioxopiperidin-3-yl)-4-methyl-1-oxo-1,2-dihydrophthalazine -6-yl)piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

3-(1-(3-(5-((1-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-5-yl )piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

3-(1-(3-(5-((1-(3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrophthalazin-6-yl )piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

3-(1-(3-(5-((1-(3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrophthalazin-5-yl )piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

3-(1-(3-(5-((1-(3-(2,6-dioxopiperidin-3-yl)-1-methyl-4-oxo-3,4-dihydrophthalazine -6-yl)piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

3-(1-(3-(5-((1-(2-(2,6-dioxopiperidin-3-yl)-7-fluoro-1-oxo-1,2-dihydrophthala zin-6-yl)piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

3-(1-(3-(5-((1-(3-(2,6-dioxopiperidin-3-yl)-7-fluoro-4-oxo-3,4-dihydrophthala zin-6-yl)piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

3-(1-(3-(5-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperidine- 4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

3-(1-(3-(5-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine- 4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

The compound according to the present invention is a compound containing a target protein ligand and an E3 enzyme ligand. Since the target protein ligand and the E3 enzyme ligand are directly covalently linked to each other without a linker, it is useful as a compound for decomposing the target protein. It can be used effectively. In particular, since the target protein ligand and the E3 enzyme ligand are directly connected without a linker, the molecular size of the compound is minimized to improve physical properties, and the protein-protein interaction between the target protein and the E3 ligand is maximized to effectively target the target protein. It is possible to disassemble.

In the present invention, the E3 enzyme ligand may be an oxophthalazine piperidinedione derivative such as the compound of formula 3 below or a dioxopiperidine isoindolinedione derivative such as the compound of formula 4 below:

[Formula 3]

In the above formula,

X is halogen or cyano (-CN),

Q ₁ to Q ₄ are each independently CH or CX,

R ¹ is hydrogen, halogen, nitro, amino, straight-chain, branched-chain or cyclic alkyl of C ₁ to C ₆ , or straight-chain, branched-chain or cyclic alkyl of C ₁ to C ₆ substituted by halogen (preferably , R ¹ is hydrogen, halogen, C ₁ to C ₄ straight chain, branched or cyclic alkyl, or C ₁ to C ₄ straight chain or cyclic alkyl substituted with halogen).

[Formula 4]

In the above formula,

Y is -C=O or -CH ₂ ,

X is halogen or cyano (-CN),

Q ₁ to Q ₄ are each independently CH or CX.

In addition, the piperidinedione derivative compound according to the present invention is a targeting protein ligand, targeting nuclear receptors (e.g., ER, AR, RAR, etc.), protein kinases (e.g., Akt, c-Abl, BTK, anaplastic lymphoma) kinase [ALK], CDK9, c-Met, RIPK2, DAPK1, PSD-95, etc.), proteins involved in transcriptional regulation (e.g., BRD4, Sirt2, HDAC6, TRIM24, IKZH1/3, Smad3, etc.), regulatory proteins (e.g. CRABP-I/II, TACC3, AHR, FKBP12, ERRα, SHP2, etc.), cellular metabolic enzymes (e.g. MetAP-2, DHODH), fusion proteins (e.g. Alk-fusion, BCR-Abl, Brd-Nut, Ret-fusion, Halo Tags, etc.), mutant proteins (e.g. For example, compounds with target proteins such as Braf mutant, EGFR mutant and deletion, Kras mutant, TP53 mutant and splicing variant, etc.) and abnormally modified proteins (e.g., EGFRdel19, P53 splicing variant, fusion proteins, etc.) can be used. However, preferably, a compound targeting c-MET protein can be used as the targeting protein ligand. Therefore, the compound according to the present invention can be usefully used in the treatment or prevention of diseases related to c-MET protein.

In the compound of the present invention, the target protein ligand is a c-MET inhibitor, and preferably a compound selected from the group consisting of the following compounds can be used:

In the present invention, the right side of the target protein ligand may be covalently bonded to any one of the carbon atoms Q1 to Q4 in the chemical formula 3 or 4, which is an E3 enzyme ligand. As such, those skilled in the art will understand that some substituents may be removed from the site where the target protein ligand and the E3 enzyme ligand are covalently linked to each other during the covalent bonding reaction.

The compound of the present invention can be usefully used in the treatment or prevention of diseases related to the target protein because the Protac compound can bind to and decompose the target protein (eg, c-MET protein). For example, the compound of the present invention has an excellent anti-cancer effect and can further induce the decomposition of c-MET protein, thereby showing a therapeutic effect on diseases related to c-MET.

Figure 1 schematically shows a Protac compound consisting of a targeting protein ligand, a linker, and an E3 enzyme ligand.
Figure 2 shows the results of an experiment confirming the proteolytic effect of the compound of the present invention in the MKN45 cell line.
Figure 3 shows the results of an experiment confirming the proteolytic effect of the compound of the present invention in SNU638, Hs746T, and EBC-1 cell lines.
Figure 4 shows the results of an experiment confirming the inhibitory effect of the compound of the present invention on proliferation of MKN45 cancer cells.
Figure 5 shows the results of an experiment confirming the inhibition of proliferation of SNU638, Hs746T, and EBC-1 cancer cells by the compound of the present invention.
Figure 6 shows the results of an experiment confirming whether the compound of the present invention has an inhibitory effect on normal cell proliferation.
Figure 7 shows the results of an experiment confirming the effect of inhibiting cell signaling at different concentrations of the compounds of the present invention.
Figure 8 shows the results of an experiment confirming the time-dependent cell signaling inhibition effect of the compound of the present invention in SNU638 cells.
Figure 9 shows the results of an experiment confirming the anticancer effect of intraperitoneal administration of the compound of the present invention in mice transplanted with SNU638 tumor cells.
Figure 10 shows the results of an experiment confirming the anticancer effect of oral administration of the compound of the present invention in mice transplanted with SNU638 tumor cells.

Hereinafter, to facilitate understanding of the present invention, the present invention will be described in detail based on examples. However, the following examples only illustrate the content of the present invention, and the spirit or scope of the present invention is not limited by the following examples in any way. Examples of the present invention are provided to more completely explain the present invention to those with average knowledge in the art.

Example A: Oxophthalazine Piperidinedione Preparation of derivative compounds

The scaffold of the oxophthalazinyl piperidione compound of Chemical Formula 1, which can be used as an E3 enzyme ligand in the Protak compound of the present invention, can be prepared through the reaction steps of Scheme 1 or Scheme 2 below. You can:

[Scheme 1]

[Scheme 2]

In Scheme 1 or 2 above, X is hydrogen, halogen, or cyano. However, for convenience of explanation, the representation of substituents that can be bonded to carbon at Q ₁ to Q ₄ in the compound of Formula 1 is omitted in the above reaction formula, and among the R ¹ substituents, only simple substituents such as hydrogen or methyl are representative. indicated.

Below, specific examples of compounds of Formula 1 are prepared through examples.

Example A1:

compound manufacture of

A1-1) Preparation of methyl 2-fluoro-6-methylbenzoate

At room temperature, to a solution of 2-fluoro-6-methylbenzoic acid (10 g, 64.9 mmol) in acetone (200 ml) was added K ₂ CO ₃ (44.8 g, 324 mmol). After stirring for 30 minutes, methane iodide (46 g, 324 mmol) was added to the reaction and heated at 60 ^o C for 6 hours. After cooling, the reaction was filtered and concentrated under reduced darkness. The product was used in the next reaction without further purification. (11.2 g, 103% yield) MS (ESI, m/z): [M+1] ⁺ = [168.3].

A1-2) Preparation of methyl 2-(bromomethyl)-6-fluorobenzoate

1-Bromopyrrolidine-2,5-dione (24.7 g, 139 mmol) in a solution of methyl 2-fluoro-6-methylbenzoate (21.2 g, 126 mmol) in ClCH ₂ CH ₂ Cl (250 ml) at room temperature. and benzoyl benzenecarboperoxoate (1.53g, 6.30mmol) were sequentially added. The reaction was heated to reflux for 16 hours. The point at which the red color disappears is the point at which the reaction is complete. After cooling, the reaction was washed with water, dried over MgSO ₄ and concentrated under reduced pressure. The crude product was used in the next reaction without further purification. (35 g, 112% yield) MS (ESI, m/z): [M+1] ⁺ = [245.9].

A1-3) Preparation of methyl 2-fluoro-6-formylbenzoate

To a solution of methyl 2-(bromomethyl)-6-fluorobenzoate (35 g, 142 mmol) in DCM (280 ml) at room temperature was added NMO (24.9 g, 212 mmol). The reaction was stirred at room temperature for 4 hours. The DCM layer was washed with water (200 ml), dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by column chromatography to give the title compound as a white solid. (11.52 g, 45% yield) MS (ESI, m/z): [M+1] ⁺ = [183.1].

A1-4) Preparation of 2-fluoro-6-formylbenzoic acid

At room temperature, an aqueous solution of lithium (+1) hydroxide (7.57 g, 180 mmol) (41 ml) was added to a solution of methyl 2-fluoro-6-formylbenzoate (11.5 g, 63.2 mmol) in THF (82 ml). . After stirring for 4 hours, the reaction was concentrated under reduced pressure and THF was dried. After cooling to 0°C, the reaction was acidified with 1N HCl to adjust the pH to 4. The reaction was extracted twice with ethyl acetate (100ml). The combined ethyl acetate layer was dried with MgSO ₄ and concentrated under reduced pressure to obtain white crystals (10.4 g, 97.8%). MS (ESI, m/z): [M+1] ⁺ = [168.8].

A1-5) Preparation of 8-fluorophthalazin-1(2H)-one

At room temperature, NH ₂ NH ₂ monohydrate (3.72 mg, 61.9 mmol) was added to a solution of 2-fluoro-6-formylbenzoic acid (10.4 g, 61.9 mmol) in methanol (120 ml) and stirred at room temperature for 16 hours. . The white precipitate was filtered and washed with methanol. The obtained white crystals were slurried with 100 ml of ethyl acetate (EA) and filtered to obtain white crystals (3.05 g, 30%). MS (ESI, m/z): [M+1] ⁺ = [164.8].

A1-6) Preparation of 3-(8-fluoro-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione

NaH (60%, 53.6 mg, 1.34 mmol) was added to a solution of 8-fluorophthalazin-1(2H)-one (200 mg, 1.22 mmol) in DMF (5 ml) at 0°C and stirred for 30 minutes. 3-Bromopiperidine-2,6-dione (468mg, 2.44mmol) was added to this solution and stirred for 6 hours. The reaction was terminated with water and extracted twice with ethyl acetate (20ml). The combined ethyl acetate was dried with MgSO ₄ and concentrated under reduced pressure. The residue was purified by column chromatography to obtain the title compound as white crystals (70 mg, 20.8%). MS (ESI, m/z): [M+1] ⁺ = [276.1].

Example A2:

compound manufacture of

A2-1) Preparation of methyl 5-fluoro-6-methylbenzoate

At room temperature, sulfuric acid (2 ml) was added to a solution of 3-fluoro-2-methylbenzoic acid (10 g, 64.9 mmol) in methanol (60 ml). Heat to 80 ^o C and stir overnight. After cooling to room temperature, the reactant was evaporated and extracted with NaHCO ₃ and ethyl acetate (EA). It was dried with MgSO ₄ . The crude product was used in the next reaction without further purification. (10.1 g, 92% yield) MS (ESI, m/z): [M+1] ⁺ = [168.3].

A2-2) Preparation of methyl 2-(bromomethyl)-3-fluorobenzoate

1-Bromopyrrolidine-2,5-dione (15.9 g, 89.2 mmol) in a solution of methyl 3-fluoro-2-methylbenzoate (10 g, 59.5 mmol) in ClCH ₂ CH ₂ Cl (250 ml) at room temperature. and benzoyl benzenecarboperoxoate (0.72g, 2.97mmol) were sequentially added. The reaction was heated to reflux for 16 hours. The point at which the red color disappears is the point at which the reaction is complete. After cooling, the reaction was washed with water, dried over MgSO ₄ and concentrated under reduced pressure. The crude product was used in the next reaction without further purification. (10.5 g, 71% yield) MS (ESI, m/z): [M+1] ⁺ = [245.9].

A2-3) Preparation of methyl 3-fluoro-2-formylbenzoate

At room temperature, to a solution of methyl 2-(bromomethyl)-3-fluorobenzoate (10 g, 40.5 mmol) in DCM (200 ml) was added NMO (10.4 g, 89 mmol), 4Å molecular sieve. The reaction was stirred at room temperature for 4 hours. The DCM layer was washed with water (200 ml), dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by column chromatography to obtain the title compound. (5.5g, 75%) MS (ESI, m/z): [M+1] ⁺ = [183.1].

A2-4) Production of 3-fluoro-2-formylbenzoic acid

An aqueous solution of lithium (+1) hydroxide monohydrate (2.88 g, 68.6 mmol) in a solution of methyl 3-fluoro-2-formylbenzoate (2.5 g, 13.7 mmol) in THF (68 ml) at room temperature (41 ml). was added. After stirring for 4 hours, the reaction was concentrated under reduced pressure and THF was dried. After cooling to 0°C, the reaction was acidified with 1N HCl to adjust the pH to 4. The reaction was extracted twice with ethyl acetate (100ml). The combined ethyl acetate layer was dried with MgSO ₄ and concentrated under reduced pressure to obtain white crystals (2.5 g, 108%). MS (ESI, m/z): [M+1] ⁺ = [168.8].

A2-5) Preparation of 5-fluoro-1,2-dihydrophthalazin-1-one

At room temperature, NH ₂ NH ₂ monohydrate (1.22 mg, 16.4 mmol) was added to a solution of 3-fluoro-2-formylbenzoic acid (2.5 g, 14.9 mmol) in THF:water=1:1 (70 ml), 16 Stirred for an hour. The mixture was acidified to pH 4 and the solid was filtered through hexane. Vacuum drying gave the title compound (1.3 g, 53%). MS (ESI, m/z): [M+1] ⁺ = [165.3]

A2-6) Preparation of 3-(5-fluoro-1-oxo-1,2-dihydrophthalazin-2-yl)piperidine-2,6-dione

LDA (lithium diisopropyl amide, 2.19 mmol) was added to a solution of 5-fluoro-1,2-dihydrophthalazin-1-one (300 mg, 1.83 mmol) in DMF (10 ml) at 0°C and stirred for 30 minutes. did. 3-Bromopiperidine-2,6-dione (526 mg, 2.74 mmol) was added to this solution and stirred at 80°C for 6 hours. When the reaction was completed, the reaction was cooled to room temperature, poured into water (100 ml), adjusted to pH 3-4 with 6N HCl, then extracted with ethyl acetate, dried over MgSO ₄ and concentrated under reduced pressure. The product was recrystallized from hexane and dried under vacuum to give the title compound. MS (ESI, m/z): [M+1] ⁺ = [276.1].

Example A3

compound manufacture of

A3-1) Preparation of methyl 4-fluoro-6-methylbenzoate

According to the preparation method of Example A2-1, methyl 4-fluoro-2-methylbenzoate was prepared from 4-fluoro-2-methylbenzoic acid.

A3-2) Preparation of methyl 2-(bromomethyl)-4-fluorobenzoate

1-Bromopyrrolidine-2,5-dione (31.8 g, 178 mmol) and benzoyl in a solution of methyl 4-fluoro-2-methylbenzoate (20 g, 119 mmol) in ClCH ₂ CH ₂ Cl (100 ml) at room temperature. Benzenecarboperoxoate (1.92g, 5.95mmol) was sequentially added. The reaction was heated to reflux for 3 hours. The point at which the red color disappears is the point at which the reaction is complete. After cooling, the reaction was washed with water, dried over MgSO ₄ and concentrated under reduced pressure. The product was purified by MPLC (HX/EA EA 0 -> 5%) (22 g, yield 75%). MS (ESI, m/z): [M+1] ⁺ = [248.4].

A3-3) Preparation of methyl 3-fluoro-2-formylbenzoate

At room temperature, NMO (15.6 mg, 134 mmol) and 4Å molecular sieves were added sequentially to a solution of methyl 2-(bromomethyl)-4-fluorobenzoate (22 g, 89 mmol) in DCM (100 ml). The reaction was stirred at room temperature for 2 hours. Molecular sieve filtered and washed with DCM (50ml). The DCM layer was washed with water (200 ml), dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by column chromatography to give the title compound as a white solid. (12g, 73.98%) MS (ESI, m/z): [M+1] ⁺ = [183.2].

A3-4) Production of 4-fluoro-2-formylbenzoic acid

At room temperature, an aqueous solution of lithium (+1) hydroxide (13.8 g, 329 mmol) (50 ml) was added to a solution of methyl 4-fluoro-6-formylbenzoate (12 g, 65.9 mmol) in THF (50 ml). After stirring for 2 hours, the reaction was concentrated under reduced pressure and THF was dried. After cooling to 0°C, the reaction was acidified with 1N HCl to adjust the pH to 4. The reaction was extracted twice with ethyl acetate (50ml). The combined ethyl acetate layer was dried with MgSO ₄ and concentrated under reduced pressure to obtain white crystals (10g, 90.3%). MS (ESI, m/z): [M+1] ⁺ = [169.2].

A3-5) Preparation of 6-fluoro-1,2-dihydrophthalazin-1-one

At room temperature, NH ₂ NH ₂ monohydrate (2.02 g, 40.3 mmol) was added to a solution of 4-fluoro-2-formylbenzoic acid (6.78 g, 40.3 mmol) in methanol (50 ml). After stirring for 30 minutes, the reaction was heated to 80° C. for 2 hours. The reaction was cooled to room temperature and concentrated under reduced pressure. The residue was poured into water (150ml) and extracted twice with ethyl acetate (150ml). The combined ethyl acetate layer was dried with MgSO ₄ and concentrated under reduced pressure to obtain white crystals (5.5 g, 83.07%). MS (ESI, m/z): [M+1] ⁺ = [165.3].

A3-6) Preparation of 3-(6-fluoro-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione

Sodium 2-methylpropane-2-oleate (175 mg, 0.914 mmol) was added to a solution of 6-fluoro-1,2-dihydrophthalazin-1-one (100 mg, 0.609 mmol) in DMF (2 ml) at 0°C. It was added. After stirring for 30 minutes, 3-bromopiperidine-2,6-dione (90 mg, 0.471 mmol) was added to the reaction mixture and stirred at room temperature for 6 hours. Water (20ml) was poured into the reaction mixture, and extracted twice with ethyl acetate (20ml). The combined ethyl acetate was dried with MgSO ₄ and concentrated under reduced pressure. The residue was purified by chromatography to obtain the title compound as white crystals (110 mg, 65.5%). MS (ESI, m/z): [M+1] ⁺ = [276.6].

Example A4

compound manufacture of

A4-1) Preparation of methyl 4-fluoro-6-methylbenzoate

According to the preparation method in Example A2, methyl 5-fluoro-2-methylbenzoate was prepared from 5-fluoro-2-methylbenzoic acid.

A4-2) Preparation of methyl 2-(bromomethyl)-5-fluorobenzoate

1-Bromopyrrolidine-2,5-dione (31.8 g, 178 mmol) and benzoyl in a solution of methyl 5-fluoro-2-methylbenzoate (20 g, 119 mmol) in ClCH ₂ CH ₂ Cl (100 ml) at room temperature. Benzenecarboperoxoate (1.92g, 5.95mmol) was sequentially added. The reaction was heated to reflux for 3 hours. The point at which the red color disappears is the point at which the reaction is complete. After cooling, the reaction was washed with water, dried over MgSO ₄ and concentrated under reduced pressure. The product was purified by MPLC (HX/EA EA 0 -> 5%) (29 g, yield 98.8%). MS (ESI, m/z): [M+1] ⁺ = [248.4].

A4-3) Preparation of methyl 5-fluoro-2-formylbenzoate

At room temperature, NMO (20.6 mg, 176 mmol) and 4Å molecular sieves were added sequentially to a solution of methyl 2-(bromomethyl)-5-fluorobenzoate (29 g, 117 mmol) in DCM (100 ml). The reaction was stirred at room temperature for 2 hours. Molecular sieve filtered and washed with DCM (50ml). The DCM layer was washed with water (200 ml), dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by column chromatography to give the title compound as a white solid. (15 g, yield 70.16%) MS (ESI, m/z): [M+1] ⁺ = [183.2].

A4-4) Production of 5-fluoro-2-formylbenzoic acid

At room temperature, an aqueous solution of lithium (+1) hydroxide (17.3 g, 412 mmol) (50 ml) was added to a solution of methyl 5-fluoro-2-formylbenzoate (15 g, 82.3 mmol) in THF (50 ml). After stirring for 2 hours, the reaction was concentrated under reduced pressure and THF was dried. After cooling to 0°C, the reaction was acidified with 1N HCl to adjust the pH to 4. The reaction was extracted twice with ethyl acetate (50ml). The combined ethyl acetate layer was dried with MgSO ₄ and concentrated under reduced pressure to obtain white crystals (12 g, 86.67%). MS (ESI, m/z): [M+1] ⁺ = [169.2].

A4-5) Preparation of 7-fluoro-1,2-dihydrophthalazin-1-one

At room temperature, NH ₂ NH ₂ monohydrate (3.74 g, 74.8 mmol) was added to a solution of 5-fluoro-2-formylbenzoic acid (8.16 g, 48.5 mmol) in methanol (50 ml). After stirring for 30 minutes, the reaction was heated to 80° C. for 2 hours. The reaction was cooled to room temperature and concentrated under reduced pressure. The residue was poured into water (150ml) and extracted twice with ethyl acetate (150ml). The combined ethyl acetate layer was dried with MgSO ₄ and concentrated under reduced pressure to obtain white crystals (6.5 g, 81.59%). MS (ESI, m/z): [M+1] ⁺ = [165.3].

A4-6) Preparation of 3-(7-fluoro-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione

Sodium 2-methylpropane-2-oleate (586 mg, 6.09 mmol) was added to a solution of 7-fluoro-1,2-dihydrophthalazin-1-one (500 mg, 3.05 mmol) in DMF (5 ml) at 0°C. It was added. After stirring for 30 minutes, 3-bromopiperidine-2,6-dione (1.05 mg, 5.48 mmol) was added to the reaction mixture and stirred at room temperature for 6 hours. The reaction mixture was poured into water (50ml) and extracted twice with ethyl acetate (50ml). The combined ethyl acetate was dried with MgSO ₄ and concentrated under reduced pressure. The residue was purified by column chromatography to obtain the title compound as white crystals (300 mg, 35.78%). MS (ESI, m/z): [M+1] ⁺ = [276.6].

Example A5

compound Preparation (prepared according to Scheme 2)

A5-1) Preparation of methyl 2-acetyl-6-fluorobenzoate

Tetrakis(triphenylphosphine) was added to a solution of methyl 2-acetyl-6-fluorobenzoate (1.12 g, 4.81 mmol) and tributyl(1-ethoxyvinyl)tin (1.91 g, 5.29 mmol) in toluene (20 ml). )-Palladium(0) (557mg, 0.48mmol) was added and stirred at 100°C for 16 hours. After cooling, 5ml of 1N-HCl was added and stirred for 1 hour. The organic layer was dried with MgSO ₄ and concentrated under reduced darkness. The residue was purified by silica column chromatography to give the title compound as a yellow oil. (820 mg, 87% yield) MS (ESI, m/z): [M+1] ⁺ = [196.8].

A5-2) Preparation of 2-acetyl-6-fluorobenzoic acid

To a solution of methyl 2-acetyl-6-fluorobenzoate (810 mg, 4.13 mmol) in THF (20 ml) and water (10 ml) was added LiOH (494 mg, 20.6 mmol) and stirred at room temperature for 20 hours. The solution was acidified with 1N-HCl to bring the pH to about 3. 100ml EA was added, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure to obtain the title compound. (810 mg yield 108%) MS (ESI, m/z): [M+1] ⁺ = [183.1].

A5-3) Preparation of 8-fluoro-4-methylphthalazin-1(2H)-one

Hydrazine monohydrate (261 mg, 5.20 mmol) was added to a solution of 2-acetyl-6-fluorobenzoic acid (790 mg, 4.34 mmol) in methanol (23 ml) and stirred at room temperature for 16 hours. The precipitate was filtered and washed with ACN (acetonitrile) to give the title compound as a white solid. (612 mg, yield 79.2%) MS (ESI, m/z): [M+1] ⁺ = [179.1].

A5-4) Preparation of 3-(8-fluoro-4-methyl-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione

To a solution of 8-fluoro-4-methylphthalazin-1(2H)-one (534 mg, 3 mmol) in THF (30 ml) at 0°C, 1M-Lithium diisopropylamide (3.6 ml, 3.6 mmol) was added, Stirred for 20 minutes. 3-Brobopiperidine-2,6-dione (863 mg, 4.5 mmol) was added to this solution and stirred at 80°C for 2 hours. The reaction product was concentrated under reduced pressure and dried. Water (10 ml) was added and stirred for 1 hour, then acidified with 1N-HCl to adjust the pH to 4. The precipitate was filtered and washed with water to give the title compound as a white solid (760 mg, yield: 86.5%). MS (ESI, m/z): [M+1] ⁺ = [289.8].

Example A6: Preparation of 3-(6-bromo-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione

A6-1) Preparation of 5-bromo-3-hydroxyisobenzofuran-1(3H)-one

To a mixture of 5-bromophthalide (5.2 g, 24.4 mmol) and N-bromosuccinimide (5.6 g, 31.7 mmol) in 200 ml ClCH ₂ CH ₂ Cl was added AIBN (401 mg, 2.44 mmol), It was refluxed for 8 hours. The reaction was followed by TLC. The succinimide was filtered off and the cake was washed with ClCH ₂ CH ₂ Cl (50 mL). The solvent was removed in vacuo, leaving 5.2 g of residue, to which 50 ml water was added. The mixture was refluxed with stirring for 4 hours, the mixture was cooled, and the product was filtered, washed for neutralization with water, and dried to give pale off-white crystals. (4.85g, yield 86.7%)

MS (ESI, m/z): [M+1] ⁺ = [229.6] and [230.5]

A6-2) Preparation of 6-bromophthalazin-1(2H)-one

To a solution of 5-bromo-3-hydroxy-1,3-dihydro-2-benzofuran-1-one (1.5 g, 6.55 mmol) in MeOH (50 mL) was added NH ² NH ₂ H ₂ O (315 mg). , 9.82 mmol) was added at room temperature and stirred for 30 minutes. The reaction was refluxed for 5 hours. The reaction was cooled to room temperature and concentrated under reduced pressure. The resulting solid was triturated with ethyl acetate to obtain white crystals. (1.31g, 5.82 mmol)

MS (ESI, m/z): [M+1] ⁺ = [226.3].

A6-3) Preparation of 3-(6-bromo-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione

To a suspension of 6-bromo-1,2-dihydrophthalazin-1-one (1.31 g, 5.82 mmol) in THF (150 mL) was added dropwise 1.0 M LDA (7.33 mL) at 0°C. After stirring for 30 minutes, 3-bromopiperidine-2,6-dione was added to the reaction mixture in portions. The reaction was heated to 80° C. and stirred for 2 hours. When the reaction was completed, the reactant was cooled to room temperature, poured into water (100 ml), adjusted to pH 3-4 with 6N HCl, extracted with ethyl acetate, dried over MgSO ₄ and concentrated under reduced pressure. The resulting solid was filtered and washed with ethyl acetate to obtain white crystals (1.73 g, 5.15 mmol).

MS (ESI, m/z): [M+1] ⁺ = [337.2].

[NMR] 1H NMR (400 MHz, DMSO-d6) δ11.08 (s, 1H), 8.45 (s, 1H), 8.28 (s, 1H), 8.18-8.16 (m, 1H), 8.06-8.04 (m , 1H), 5.84-5.80 (m, 1H), 2.93 - 2.90 (m, 1H), 2.65 - 2.54 (m, 2H), 2.15 - 2.12 (m, 1H).

Example B: (2S)-2-{[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazine-1 -yl]methyl}-4-(5-{4-[(piperazin-1-yl)methyl]phenyl}pyrimidin-2-yl)morpholine synthesis

A c-MET target protein ligand compound of the following formula can be prepared through the following reaction process:

Step 1) Synthesis of t-butyl (2S)-2-{[(4-methylbenzenesulfonyl)oxy]methyl}morpholine-4-carboxylate

To a solution of t-butyl (2S)-2-(hydroxymethyl)morpholine-4-carboxylate (10 g, 46 mmol) in DCM (500 ml) was added 4-methylbenzene-1-sulfonyl chloride (9.65 g, 50.6 mmol), N,N-dimethylpyridin-4-amine (0.84 g, 6.9 mmol) and triethylamine (3.0 equivalents) were added. The reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reactant was poured into water, extracted with DCM (twice), dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by hexane/ethyl acetate (0~30%) column chromatography to obtain the title compound (17 g).

MS (ESI, m/z): [M+H] ⁺ = 372.4.

Step 2) Synthesis of t-butyl (2S)-2-(azidomethyl)morpholine-4-carboxylate

To a solution of t-butyl (2S)-2-{[(4-methylbenzenesulfonyl)oxy]methyl}morpholine-4-carboxylate (17 g, 45.8 mmol) in DMF (500 ml) was added sodium azide (12). g, 183 mmol) and sodium iodide (1.03 g, 6.86 mmol) were added. The reaction was stirred at 70° C. for 12 hours. After completion of the reaction, the reaction was poured into water, extracted with ethyl acetate (twice), dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by hexane/ethyl acetate (0~30%) column chromatography to obtain the title compound (10.2 g).

MS (ESI, m/z): [M+H] ⁺ = 243.4.

Step 3) Synthesis of t-butyl (2R)-2-(aminomethyl)morpholine-4-carboxylate

To a solution of t-butyl (2S)-2-(azidomethyl)morpholine-4-carboxylate (9.5 g, 39.2 mmol) in MeOH (500 ml) was added Pd/C (2 g, 5%), then Reacted for 1 hour under H ₂ (gas). After completion of the reaction, the reaction mixture was filtered through Celite and concentrated under reduced pressure. The title compound (8.4 g) was used directly in the next reaction without further purification.

MS (ESI, m/z): [M+H] ⁺ = 217.4.

Step 4) Synthesis of t-butyl (2R)-2-{[(3-amino-6-bromopyrazin-2-yl)amino]methyl}morpholine-4-carboxylate

t-Butyl (2R)-2-(aminomethyl)morpholine-4-carboxylate (8.4 g, 38.8 mmol) and 3,5-dibromopyrazin-2-amine (11.8 g, Triethylamine (3.0 equivalents) was added to the solution (46.6 mmol). The reaction was stirred at 150° C. for 12 hours. After completion of the reaction, the reaction was poured into water, extracted with ethyl acetate (twice), dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by hexane/ethyl acetate (20-50%) column chromatography to obtain the title compound (8.5 g).

MS (ESI, m/z): [M+H] ⁺ = 389.4.

Step 5) t-Butyl (2S)-2-({5-bromo-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl}methyl)morpholine-4- Carboxylate synthesis

t-Butyl (2R)-2-{[(3-amino-6-bromopyrazin-2-yl)amino]methyl}morpholine-4-carboxylate (8.4 g, 21.6 mmol) in DMF (300 ml) Isoamyl nitrite (3.8 g, 32.5 mmol) was added to the solution. The reaction was stirred at 70° C. for 3 hours. After completion of the reaction, the reaction was poured into water, extracted with ethyl acetate (twice), dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by hexane/ethyl acetate (0~30%) column chromatography to obtain the title compound (7.5g).

MS (ESI, m/z): [M+H] ⁺ = 400.4.

Step 6) t-Butyl (2S)-2-{[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazine -1-yl]methyl}morpholine-4-carboxylate synthesis

t-Butyl (2S)-2-({5-bromo-1H-[1,2,3]triazolo[4,5) in 1,4-dioxane (300 ml) and H ₂ O (100 ml) -b]pyrazin-1-yl}methyl)morpholine-4-carboxylate (7.5 g, 18.8 mmol), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2- Pd(dppf)Cl2 (0.7 g, 0.95 mmol) was added to a solution of dioxaborolan-2-yl)-1H-pyrazole (4.7 g, 22.5 mmol) and cesium carbonate (18.4 g, 56.4 mmol). The reaction was stirred at 80° C. for 2 hours. After completion of the reaction, the reaction was poured into water, extracted with ethyl acetate (twice), dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by hexane/ethyl acetate (10-30%) column chromatography to obtain the title compound (7.1 g).

MS (ESI, m/z): [M+H] ⁺ = 401.4.

Step 7) (2S)-2-{[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazine-1- [1]methyl}morpholine synthesis

t-Butyl (2S)-2-{[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5- in DCM (300 ml) To a solution of b]pyrazin-1-yl]methyl}morpholine-4-carboxylate (7.1 g, 17.7 mmol), TFA (100 ml) was added. The reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The title compound (4.8 g) was used directly in the next reaction without further purification.

MS (ESI, m/z): [M+H] ⁺ = 301.4.

Step 8) (2S)-4-(5-bromopyrimidin-2-yl)-2-{[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2, 3]triazolo[4,5-b]pyrazin-1-yl]methyl}morpholine synthesis

(2S)-2-{[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazine in EtOH (300 ml) -1-yl]methyl}morpholine (4.8 g, 16 mmol) and 5-bromo-2-chloropyrimidine (4.64 g, 24 mmol) in a solution of ethylbis(propan-2-yl)amine (3.0 equiv) was added. The reaction was stirred at 50° C. for 9 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The title compound was recrystallized from EtOH. The title compound (5.5 g) was used directly in the next reaction without further purification.

MS (ESI, m/z): [M+H] ⁺ = 458.4.

Step 9) 4-{2-[(2S)-2-{[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5- b]pyrazin-1-yl]methyl}morpholin-4-yl]pyrimidin-5-yl}benzaldehyde synthesis

(2S)-4-(5-bromopyrimidin-2-yl)-2-{[5-(1-methyl-1H) in 1,4-dioxane (150 ml) and H ₂ O (50 ml) -pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl]methyl}morpholine (5.5 g, 12 mmol), 4-(4, Pd(dppf)Cl2 in a solution of 4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde (4.2 g, 18 mmol) and cesium carbonate (8.31 g, 60.1 mmol). (1.8 g, 2.41 mmol) was added. The reaction was stirred at 90° C. for 2 hours. After completion of the reaction, the reaction was poured into water, extracted with ethyl acetate (twice), dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by hexane/ethyl acetate (30-50%) column chromatography to obtain the title compound (5 g).

MS (ESI, m/z): [M+H] ⁺ = 483.4.

Step 10) t-Butyl 4-[(4-{2-[(2S)-2-{[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3] Synthesis of triazolo[4,5-b]pyrazin-1-yl]methyl}morpholin-4-yl]pyrimidin-5-yl}phenyll)methyl]piperazine-1-carboxylate

4-{2-[(2S)-2-{[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4 in DCM (250 ml) ,5-b]pyrazin-1-yl]methyl}morpholin-4-yl]pyrimidin-5-yl}benzaldehyde (5 g, 10.4 mmol) and t-butyl piperazine-1-carboxylate (4.8 g , 26 mmol), sodium triacetoxyborohydride (3.9 g, 18.7 mmol) and AcOH (1 ml) were added to the solution. The reaction was stirred at room temperature for 18 hours. After completion of the reaction, the reaction was poured into water, extracted with DCM (twice), dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by dichloromethane/methanol (0~10%) column chromatography to obtain the title compound (6.7g).

MS (ESI, m/z): [M+H] ⁺ = 653.8.

Step 11) (2S)-2-{[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazine-1- 1]methyl}-4-(5-{4-[(piperazin-1-yl)methyl]phenyl}pyrimidin-2-yl)morpholine synthesis

t-Butyl 4-[(4-{2-[(2S)-2-{[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2 in DCM (150 ml) ,3]triazolo[4,5-b]pyrazin-1-yl]methyl}morpholin-4-yl]pyrimidin-5-yl}phenyll)methyl]piperazine-1-carboxylate (6.7 g, 10.3 mmol) TFA (50 ml) was added to the solution. The reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The residue was purified by dichloromethane/methanol (0~10%) column chromatography to obtain the title compound (6.7g).

MS (ESI, m/z): [M+H] ⁺ = 553.6.

Example C: Preparation of compounds for TPD

Example 1:

3-(6-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]tria Zolo[4,5-b]pyrazin-1-yl)methyl)morpholino)pyrimidin-5-yl)benzyl)piperazin-1-yl)-1-oxophthalazin-2(1H)-yl ) piperidine-2,6-dione

3-(6-fluoro-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione (50 mg) in NMP (N-methyl-2-pyrrolidone) (3 ml) , 0.182 mmol) and (S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazine- 1-yl)methyl)-4-(5-(4-(piperazin-1-ylmethyl)phenyl)pyrimidin-2-yl)morpholine (100 mg, 0.182 mmol) in a solution of ethylbis(propane-2) -yl)amine (3.0 equiv) was added to the reaction mixture. The reaction was stirred at 130° C. for 24 hours. The residue was purified by MeOH/DCM (0-20%) MPLC (medium pressure liquid chromatography) to obtain the title compound (70 mg).

MS (ESI, m/z): [M+H] ⁺ = 808.8

¹ H NMR (500 MHz, DMSO-d 6 ) d ppm 11.01 (s, 1 H) 9.21 (s, 1 H) 8.72 - 8.80 (m, 2 H) 8.64 (s, 1 H) 8.31 (s, 1 H) ) 8.22 - 8.27 (m, 1 H) 8.05 (t, J=8.32 Hz, 1 H) 7.59 - 7.69 (m, 2 H) 7.46 - 7.54 (m, 1 H) 7.34 - 7.46 (m, 1 H) 7.26 (d, J=5.49 Hz, 1 H) 5.75 (d, J=6.87 Hz, 1 H) 4.86 - 5.00 (m, 3 H) 4.79 (d, J=10.68 Hz, 2 H) 4.73 (d, J= 12.66 Hz, 2 H) 4.39 (d, J=12.51 Hz, 1 H) 4.22 (br. s., 1 H) 3.88 - 3.98 (m, 4 H) 3.64 (s, 1 H) 3.39 - 3.62 (m, 7 H) 3.07 - 3.17 (m, 2 H) 2.85 - 2.98 (m, 2 H) 2.14 - 2.21 (m, 2 H).

Example 2:

3-(4-methyl-6-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2 ,3] triazolo [4,5-b] pyrazin-1-yl) methyl) morpholino) pyrimidin-5-yl) benzyl) piperazin-1-yl) -1-oxophthalazine-2 ( 1H)-yl)piperidine-2,6-dione

3-(6-fluoro-4-methyl-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione (50 mg, 0.182 mmol) and (2S) in NMP (3 ml) )-2-{[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl]methyl}- 4-(5-{4-[(piperazin-1-yl)methyl]phenyl}pyrimidin-2-yl)morpholine (100 mg, 0.182 mmol) was added to a solution of ethylbis(propan-2-yl)amine ( 3.0 equivalent) was added to the reaction mixture. The reaction was stirred at 130° C. for 24 hours. The residue was purified by MPLC with MeOH/DCM (0~20%) to obtain the title compound (75 mg).

MS (ESI, m/z): [M+H] ⁺ = 822.92

¹ H NMR (500 MHz, DMSO-d 6 ) d ppm 10.98 (s, 1 H) 9.21 (s, 2 H) 8.78 (s, 2 H) 8.65 (s, 2 H) 8.31 (s, 2 H) 7.86 - 8.03 (m, 1 H) 7.50 (d, J=8.70 Hz, 1 H) 4.86 - 4.99 (m, 3 H) 4.73 (d, J=12.05 Hz, 1 H) 4.39 (d, J=13.73 Hz, 1 H) 3.90 - 3.98 (m, 9 H) 3.36 - 3.54 (m, 11 H) 3.07 - 3.17 (m, 2 H) 2.53 - 2.66 (m, 6 H).

Example 3:

3-(7-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]tria Zolo[4,5-b]pyrazin-1-yl)methyl)morpholino)pyrimidin-5-yl)benzyl)piperazin-1-yl)-1-oxophthalazin-2(1H)-yl ) piperidine-2,6-dione

3-(7-Fluoro-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione (50 mg, 0.182 mmol) and (2S)-2- in NMP (3 ml) {[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl]methyl}-4-(5 -{4-[(piperazin-1-yl)methyl]phenyl}pyrimidin-2-yl)morpholine (100 mg, 0.182 mmol) was reacted with ethylbis(propan-2-yl)amine (3.0 equivalents). added to the mixture. The reaction was stirred at 130° C. for 24 hours. The residue was purified by MeOH/DCM (0~20%) MPLC to obtain the title compound (60 mg).

MS (ESI, m/z): [M+H] ⁺ = 808.8

¹ H NMR (500 MHz, DMSO-d 6 ) d ppm 10.92 (s, 1 H) 9.21 (s, 2 H) 8.88 (s, 1 H) 8.73 - 8.78 (m, 1 H) 8.62 - 8.68 (m, 2 H) 8.29 - 8.35 (m, 2 H) 8.26 (d, J=5.95 Hz, 2 H) 7.91 - 8.00 (m, 1 H) 7.43 (d, J=8.09 Hz, 1 H) 4.81 - 4.99 (m , 2 H) 3.88 - 3.96 (m, 9 H) 3.41 - 3.51 (m, 1 H) 3.29 - 3.47 (m, 11 H) 3.06 - 3.18 (m, 2 H) 2.53 - 2.68 (m, 3 H).

Example 4:

3-(8-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]tria Zolo[4,5-b]pyrazin-1-yl)methyl)morpholino)pyrimidin-5-yl)benzyl)piperazin-1-yl)-1-oxophthalazin-2(1H)-yl ) piperidine-2,6-dione

3-(8-Fluoro-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione (50 mg, 0.182 mmol) and (2S)-2- in NMP (3 ml) {[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl]methyl}-4-(5 -{4-[(piperazin-1-yl)methyl]phenyl}pyrimidin-2-yl)morpholine (100 mg, 0.182 mmol) was reacted with ethylbis(propan-2-yl)amine (3.0 equivalents). added to the mixture. The reaction was stirred at 130° C. for 24 hours. The residue was purified by MPLC with MeOH/DCM (0~20%) to obtain the title compound (53 mg).

MS (ESI, m/z): [M+H] ⁺ = 808.8

¹ H NMR (500 MHz, DMSO-d 6 ) d ppm 10.92 (s, 1 H) 9.14 (s, 1 H) 8.69 (s, 2 H) 8.57 (s, 1 H) 8.24 (s, 1 H) 8.21 (s, 1 H) 7.69 - 7.75 (m, 1 H) 7.56 (d, J=8.09 Hz, 2 H) 7.31 - 7.39 (m, 2 H) 7.26 (d, J=8.24 Hz, 1 H) 4.79 - 4.93 (m, 2 H) 4.66 (d, J=12.51 Hz, 1 H) 4.30 (br. s., 1 H) 3.82 - 3.90 (m, 4 H) 3.28 - 3.45 (m, 14 H) 2.99 - 3.10 (m, 3 H) 2.55 (br. s., 3 H).

Example 5:

3-(4-methyl-8-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2 ,3] triazolo [4,5-b] pyrazin-1-yl) methyl) morpholino) pyrimidin-5-yl) benzyl) piperazin-1-yl) -1-oxophthalazine-2 ( 1H)-yl)piperidine-2,6-dione

3-(8-fluoro-4-methyl-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione (50 mg, 0.182 mmol) and (2S) in NMP (3 ml) )-2-{[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl]methyl}- 4-(5-{4-[(piperazin-1-yl)methyl]phenyl}pyrimidin-2-yl)morpholine (100 mg, 0.182 mmol) was added to a solution of ethylbis(propan-2-yl)amine ( 3.0 equivalent) was added to the reaction mixture. The reaction was stirred at 130° C. for 24 hours. The residue was purified by MPLC with MeOH/DCM (0~20%) to obtain the title compound (45 mg).

MS (ESI, m/z): [M+H] ⁺ = 822.92

¹ H NMR (500 MHz, DMSO-d 6 ) d ppm 10.92 (s, 1 H) 9.14 (s, 1 H) 8.74 (s, 2 H) 8.58 (s, 1 H) 8.24 (s, 1 H) 7.80 (t, J=8.09 Hz, 1 H) 7.73 (d, J=7.02 Hz, 1 H) 7.58 (br. s., 1 H) 7.48 (d, J=7.93 Hz, 1 H) 7.35 (d, J =8.39 Hz, 1 H) 4.79 - 4.93 (m, 2 H) 4.67 (d, J=12.82 Hz, 1 H) 4.33 (d, J=13.28 Hz, 1 H) 3.82 - 3.91 (m, 4 H) 3.29 - 3.47 (m, 14 H) 3.01 - 3.11 (m, 2 H) 2.78 - 2.90 (m, 1 H) 2.46 - 2.63 (m, 3 H) 2.43 (s, 3H).

Example 6:

3-(6-fluoro-7-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1, 2,3]triazolo[4,5-b]pyrazin-1-yl)methyl)morpholino)pyrimidin-5-yl)benzyl)piperazin-1-yl)-1-oxophthalazine-2 (1H)-yl)piperidine-2,6-dione

3-(6,7-difluoro-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione (50 mg, 0.182 mmol) and (2S)- in NMP (3 ml) 2-{[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl]methyl}-4- (5-{4-[(piperazin-1-yl)methyl]phenyl}pyrimidin-2-yl)morpholine (100 mg, 0.182 mmol) was added to a solution of ethylbis(propan-2-yl)amine (3.0 equivalents) ) was added to the reaction mixture. The reaction was stirred at 130° C. for 24 hours. The residue was purified by MPLC with MeOH/DCM (0~20%) to obtain the title compound (50 mg).

MS (ESI, m/z): [M+H] ⁺ = 826.82

¹ H NMR (500 MHz, DMSO-d 6 ) d ppm 10.98 (s, 1 H) 9.14 (s, 1 H) 8.74 (s, 2 H) 8.58 (s, 1 H) 8.26 - 8.35 (m, 1 H) ) 8.24 (s, 1 H) 7.81 - 7.91 (m, 1 H) 7.71 - 7.81 (m, 2 H) 7.67 (d, J=8.39 Hz, 1 H) 7.55 (br. s., 3 H) 5.72 ( br. s., 1 H) 4.78 - 4.92 (m, 2 H) 4.67 (d, J=12.51 Hz, 1 H) 4.40 (br. s., 1 H) 4.33 (d, J=13.28 Hz, 1 H ) 4.15 (br. s., 1 H) 3.79 - 3.92 (m, 4 H) 3.73 (br. s., 2 H) 3.39 - 3.51 (m, 3 H) 3.18 (br. s., 3 H) 3.01 - 3.13 (m, 3 H) 2.76 - 2.92 (m, 1 H) 2.45 - 2.59 (m, 3 H).

Example 7:

2-(2,6-dioxopiperidin-3-yl)-4-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazole-4 -yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl)methyl)morpholino)pyrimidin-5-yl)benzyl)piperazin-1-yl) Isoindoline-1,3-dione

2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (50 mg, 0.182 mmol) and (2S)-2-{[5 -(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl]methyl}-4-(5-{4 -[(piperazin-1-yl)methyl]phenyl}pyrimidin-2-yl)morpholine (100 mg, 0.182 mmol) was added to the reaction mixture of ethylbis(propan-2-yl)amine (3.0 equivalents). It was added. The reaction was stirred at 130° C. for 24 hours. The residue was purified by MPLC with MeOH/DCM (0~20%) to obtain the title compound (80 mg).

MS (ESI, m/z): [M+H] ⁺ = 809.82

¹ H NMR (500 MHz, DMSO-d 6 ) d ppm 11.03 (s, 1 H) 9.14 (s, 1 H) 8.70 (s, 2 H) 8.58 (s, 1 H) 8.24 (s, 1 H) 7.64 (t, J=7.63 Hz, 1 H) 7.58 (br. s., 1 H) 7.37 (br. s., 2 H) 7.23 - 7.34 (m, 2 H) 5.02 (dd, J=12.66, 5.34 Hz , 1 H) 4.78 - 4.96 (m, 2 H) 4.66 (d, J=12.51 Hz, 2 H) 4.32 (d, J=13.12 Hz, 1 H) 4.15 (br. s., 1 H) 3.80 - 3.92 (m, 4 H) 3.53 (br. s., 1 H) 3.37 - 3.49 (m, 2 H) 3.33 (br. s., 1 H) 2.98 - 3.13 (m, 2 H) 2.71 - 2.88 (m, 1 H) 2.46 - 2.65 (m, 5 H) 1.79 - 2.00 (m, 3 H) 1.68 (br. s., 1 H) 1.38 (br. s., 1 H).

Example 8:

2-(2,6-dioxopiperidin-3-yl)-5-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazole-4 -yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl)methyl)morpholino)pyrimidin-5-yl)benzyl)piperazin-1-yl) Isoindoline-1,3-dione

2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (50 mg, 0.182 mmol) and (2S)-2-{[5 -(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl]methyl}-4-(5-{4 -[(piperazin-1-yl)methyl]phenyl}pyrimidin-2-yl)morpholine (100 mg, 0.182 mmol) was added to the reaction mixture of ethylbis(propan-2-yl)amine (3.0 equivalents). It was added. The reaction was stirred at 130° C. for 24 hours. The residue was purified by MPLC with MeOH/DCM (0~20%) to obtain the title compound (80 mg).

MS (ESI, m/z): [M+H] ⁺ = 809.82

¹ H NMR (500 MHz, DMSO-d 6 ) d ppm 11.02 (s, 1 H) 9.14 (s, 1 H) 8.63 - 8.71 (m, 1 H) 8.54 - 8.60 (m, 1 H) 8.24 (s, 1 H) 7.66 - 7.77 (m, 1 H) 7.62 (d, J=8.54 Hz, 1 H) 7.56 (br. s., 1 H) 7.37 (br. s., 1 H) 7.28 (br. s. , 1 H) 7.19 (d, J=8.85 Hz, 1 H) 4.96 - 5.05 (m, 1 H) 4.78 - 4.93 (m, 2 H) 4.62 - 4.76 (m, 2 H) 4.32 (d, J=12.51 Hz, 1 H) 4.15 (br. s., 1 H) 3.79 - 3.90 (m, 4 H) 3.50 (br. s., 1 H) 3.31 - 3.48 (m, 3 H) 2.97 - 3.13 (m, 3) H) 2.80 (d, J=12.51 Hz, 2 H) 2.50 - 2.61 (m, 5 H) 1.77 - 1.96 (m, 3 H) 1.69 (br. s., 1 H) 1.38 (br. s., 1 H).

Example 9:

3-(1-(3-(5-((1-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl )piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

3-(6-fluoro-1-oxo-1,2-dihydrophthalazin-2-yl)piperidine-2,6-dione (50 mg, 0.182 mmol) and (3) in NMP (3 ml) -(6-oxo-1-(3-(5-(piperidin-4-ylmethoxy)pyrimidin-2-yl)benzyl)-1,6-dihydropyridazin-3-yl)benzonitrile ( 120 mg, 0.182 mmol) ethylbis(propan-2-yl)amine (3.0 equivalents) was added to the reaction mixture. The reaction was stirred at 130° C. for 24 hours. The residue was dissolved in MeOH/DCM (0-20%). ) Purified by MPLC to give the title compound (90 mg).

MS (ESI, m/z): [M+H] ⁺ =734.80

¹ H NMR (500 MHz, DMSO-d 6 ) d ppm 10.93 (s, 1 H) 8.59 (s, 2 H) 8.31 (s, 2 H) 8.15 - 8.21 (m, 3 H) 8.11 (d, J= 9.77 Hz, 1 H) 7.96 (d, J=9.00 Hz, 1 H) 7.87 (d, J=7.63 Hz, 1 H) 7.65 (t, J=7.86 Hz, 1 H) 7.37 - 7.51 (m, 3 H) ) 7.16 - 7.25 (m, 1 H) 7.10 (d, J=9.77 Hz, 1 H) 5.68 (dd, J=11.67, 5.11 Hz, 1 H) 5.38 (s, 2 H) 3.96 - 4.09 (m, 4 H) 2.77 - 3.00 (m, 3 H) 2.45 - 2.59 (m, 1 H) 2.07 (br. s., 1 H) 1.97 - 2.05 (m, 1 H) 1.84 (d, J=11.44 Hz, 2 H ) 1.26 - 1.43 (m, 2 H).

Example 10:

3-(1-(3-(5-((1-(2-(2,6-dioxopiperidin-3-yl)-4-methyl-1-oxo-1,2-dihydrophthalazine -6-yl)piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

3-(6-fluoro-4-methyl-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione (50 mg, 0.182 mmol) and (3) in NMP (3 ml) -(6-oxo-1-(3-(5-(piperidin-4-ylmethoxy)pyrimidin-2-yl)benzyl)-1,6-dihydropyridazin-3-yl)benzonitrile ( 120 mg, 0.182 mmol) ethylbis(propan-2-yl)amine (3.0 equivalents) was added to the reaction mixture. The reaction was stirred at 130° C. for 24 hours. The residue was dissolved in MeOH/DCM (0-20%). ) Purified by MPLC to give the title compound (70 mg).

MS (ESI, m/z): [M+H] ⁺ = 748.81 ¹ H NMR (500 MHz, DMSO-d 6 ) d ppm 10.91 (s, 1 H) 8.52 - 8.64 (m, 2 H) 8.25 - 8.34 (m, 2 H) 8.08 - 8.22 (m, 3 H) 7.99 (d, J=9.00 Hz, 1 H) 7.86 (d, J=7.63 Hz, 1 H) 7.65 (t, J=7.86 Hz, 1 H ) 7.33 - 7.48 (m, 3 H) 7.09 (d, J=9.77 Hz, 1 H) 6.97 - 7.05 (m, 1 H) 5.57 - 5.67 (m, 1 H) 5.32 - 5.43 (m, 3 H) 4.00 - 4.13 (m, 4 H) 2.77 - 3.00 (m, 3 H) 2.40 - 2.60 (m, 5 H) 2.03 - 2.11 (m, 1 H) 1.94 - 2.03 (m, 1 H) 1.84 (d, J= 11.44 Hz, 2 H) 1.36 (q, J=11.44 Hz, 2 H).

Example 11:

3-(1-(3-(5-((1-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-5-yl )piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

3-(5-fluoro-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione (50 mg, 0.182 mmol) and (3-(6-) in NMP (3 ml) Oxo-1-(3-(5-(piperidin-4-ylmethoxy)pyrimidin-2-yl)benzyl)-1,6-dihydropyridazin-3-yl)benzonitrile (120 m g, 0.182 mmol) Ethylbis(propan-2-yl)amine (3.0 equivalents) was added to the reaction mixture. The reaction was stirred at 130° C. for 24 hours. The residue was purified by MeOH/DCM (0-20%) MPLC. Thus, the title compound (90 mg) was obtained.

MS (ESI, m/z): [M+H] ⁺ =734.80

¹ H NMR (500 MHz, DMSO-d 6 ) d ppm 10.99 (s, 1 H) 8.58 - 8.66 (m, 2 H) 8.29 - 8.38 (m, 3 H) 8.15 - 8.22 (m, 2 H) 8.11 ( d, J=9.77 Hz, 1 H) 7.84 (d, J=7.93 Hz, 1 H) 7.87 (d, J=7.63 Hz, 1 H) 7.72 (t, J=7.86 Hz, 1 H) 7.65 (t, J=7.86 Hz, 1 H) 7.49 (d, J=7.93 Hz, 1 H) 7.38 - 7.44 (m, 2 H) 7.10 (d, J=9.77 Hz, 1 H) 5.74 (dd, J=11.52, 4.65 Hz, 1 H) 5.38 (s, 2 H) 4.10 (d, J=6.26 Hz, 2 H) 2.73 - 2.93 (m, 3 H) 2.45 - 2.60 (m, 3 H) 2.00 - 2.13 (m, 2 H) ) 1.85 - 1.99 (m, 4 H) 1.60 (q, J=11.29 Hz, 2 H).

Example 12:

3-(1-(3-(5-((1-(3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrophthalazin-6-yl )piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

3-(7-fluoro-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione (50 mg, 0.182 mmol) and (3-(6-) in NMP (3 ml) Oxo-1-(3-(5-(piperidin-4-ylmethoxy)pyrimidin-2-yl)benzyl)-1,6-dihydropyridazin-3-yl)benzonitrile (120 mg, 0.182 mmol) Ethylbis(propan-2-yl)amine (3.0 equivalents) was added to the reaction mixture. The reaction was stirred at 130° C. for 24 hours. The residue was purified by MeOH/DCM (0-20%) MPLC. Thus, the title compound (40 mg) was obtained.

MS (ESI, m/z): [M+H] ⁺ =734.80

¹ H NMR (500 MHz, DMSO-d 6 ) d ppm 10.94 (s, 1 H) 8.54 - 8.64 (m, 2 H) 8.31 (d, J=1.68 Hz, 2 H) 8.13 - 8.21 (m, 3 H) ) 8.11 (d, J=9.77 Hz, 1 H) 7.86 (d, J=7.78 Hz, 1 H) 7.70 (d, J=9.00 Hz, 1 H) 7.65 (t, J=7.86 Hz, 1 H) 7.56 (dd, J=9.00, 2.44 Hz, 1 H) 7.36 - 7.48 (m, 3 H) 7.09 (d, J=9.77 Hz, 1 H) 5.63 - 5.74 (m, 1 H) 5.38 (s, 2 H) 3.95 - 4.08 (m, 4 H) 2.78 - 2.97 (m, 3 H) 2.45 - 2.62 (m, 3 H) 1.98 - 2.12 (m, 2 H) 1.85 (d, J=11.44 Hz, 2 H) 1.35 ( q, J=11.19 Hz, 2 H).

Example 13:

3-(1-(3-(5-((1-(3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrophthalazin-5-yl )piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

3-(8-fluoro-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione (50 mg, 0.182 mmol) and (3-(6-) in NMP (3 ml) Oxo-1-(3-(5-(piperidin-4-ylmethoxy)pyrimidin-2-yl)benzyl)-1,6-dihydropyridazin-3-yl)benzonitrile (120 mg, 0.182 mmol) Ethylbis(propan-2-yl)amine (3.0 equivalents) was added to the reaction mixture. The reaction was stirred at 130° C. for 24 hours. The residue was purified by MeOH/DCM (0-20%) MPLC. Thus, the title compound (40 mg) was obtained.

MS (ESI, m/z): [M+H] ⁺ =734.80

¹ H NMR (500 MHz, DMSO-d 6 ) d ppm10.93 (s, 1 H) 8.56 - 8.66 (m, 2 H) 8.32 (d, J=6.10 Hz, 2 H) 8.14 - 8.24 (m, 3 H) 8.11 (d, J=9.77 Hz, 1 H) 7.87 (d, J=7.63 Hz, 1 H) 7.72 (t, J=7.93 Hz, 1 H) 7.65 (t, J=7.86 Hz, 1 H) 7.37 - 7.47 (m, 2 H) 7.24 - 7.37 (m, 2 H) 7.10 (d, J=9.77 Hz, 1 H) 5.38 (s, 2 H) 4.08 (d, J=6.26 Hz, 2 H) 3.38 (d, J=9.77 Hz, 2 H) 2.79 - 2.92 (m, 1 H) 2.69 (q, J=10.58 Hz, 2 H) 2.45 - 2.60 (m, 4 H) 1.98 - 2.09 (m, 1 H) 1.90 (br. s., 1 H) 1.80 (br. s., 2 H) 1.52 - 1.68 (m, 2 H).

Example 14:

3-(1-(3-(5-((1-(3-(2,6-dioxopiperidin-3-yl)-1-methyl-4-oxo-3,4-dihydrophthalazine -6-yl)piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

3-(7-Fluoro-4-methyl-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione (50 mg, 0.182 mmol) and (3) in NMP (3 ml) -(6-oxo-1-(3-(5-(piperidin-4-ylmethoxy)pyrimidin-2-yl)benzyl)-1,6-dihydropyridazin-3-yl)benzonitrile ( 120 mg, 0.182 mmol) ethylbis(propan-2-yl)amine (3.0 equivalents) was added to the reaction mixture. The reaction was stirred at 130° C. for 24 hours. The residue was dissolved in MeOH/DCM (0-20%). ) Purified by MPLC to give the title compound (38 mg).

MS (ESI, m/z): [M+H] ⁺ = 748.81

¹ H NMR (500 MHz, DMSO-d 6 ) d ppm 10.91 (s, 1 H) 8.59 (s, 2 H) 8.25 - 8.34 (m, 2 H) 8.13 - 8.21 (m, 2 H) 8.11 (d, J=9.77 Hz, 1 H) 7.86 (d, J=7.78 Hz, 1 H) 7.70 (d, J=9.00 Hz, 1 H) 7.65 (t, J=7.86 Hz, 1 H) 7.54 (dd, J= 9.08, 2.37 Hz, 1 H) 7.47 (d, J=2.44 Hz, 1 H) 7.37 - 7.45 (m, 2 H) 7.10 (d, J=9.77 Hz, 1 H) 5.63 (d, J=6.10 Hz, 1 H) 5.38 (s, 2 H) 3.94 - 4.09 (m, 4 H) 2.77 - 3.00 (m, 3 H) 2.45 - 2.61 (m, 4 H) 2.39 (s, 3 H) 1.97 - 2.06 (m, 1 H) 1.85 (d, J=12.21 Hz, 2 H) 1.26 - 1.43 (m, 2 H).

Example 15:

3-(1-(3-(5-((1-(2-(2,6-dioxopiperidin-3-yl)-7-fluoro-1-oxo-1,2-dihydrophthala zin-6-yl)piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile and 3-( 1-(3-(5-((1-(3-(2,6-dioxopiperidin-3-yl)-7-fluoro-4-oxo-3,4-dihydrophthalazine-6 -yl)piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile)

3-(6,7-difluoro-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione (50 mg, 0.182 mmol) and (3- (6-oxo-1-(3-(5-(piperidin-4-ylmethoxy)pyrimidin-2-yl)benzyl)-1,6-dihydropyridazin-3-yl)benzonitrile (120 mg, 0.182 mmol) ethylbis(propan-2-yl)amine (3.0 equivalents) was added to the reaction mixture. The reaction was stirred at 130° C. for 24 hours. The residue was dissolved in MeOH/DCM (0-20%). Purification by MPLC gave the title compound (50 mg).

MS (ESI, m/z): [M+H] ⁺ = 752.78

Example 16:

3-(1-(3-(5-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperidine- 4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (50 mg, 0.182 mmol) and (3-(6-oxo-) in NMP (3 ml) 1-(3-(5-(piperidin-4-ylmethoxy)pyrimidin-2-yl)benzyl)-1,6-dihydropyridazin-3-yl)benzonitrile (120 mg, 0.182 mmol) To the solution, ethylbis(propan-2-yl)amine (3.0 equivalents) was added to the reaction mixture.The reaction was stirred for 24 hours at 130° C. The residue was purified by MeOH/DCM (0~20%) MPLC, The title compound (80 mg) was obtained.

MS (ESI, m/z): [M+H] ⁺ = 735.75

¹ H NMR (500 MHz, DMSO-d 6 ) d ppm 11.10 (s, 1 H) 8.68 (s, 2 H) 8.39 (d, J=5.95 Hz, 2 H) 8.22 - 8.28 (m, 2 H) 8.18 (d, J=9.77 Hz, 1 H) 7.94 (d, J=7.78 Hz, 1 H) 7.67 - 7.76 (m, 2 H) 7.46 - 7.52 (m, 2 H) 7.34 (d, J=7.17 Hz, 1 H) 7.37 (d, J=8.54 Hz, 1 H) 7.17 (d, J=9.77 Hz, 1 H) 5.45 (s, 2 H) 5.10 (dd, J=12.82, 5.49 Hz, 1 H) 4.15 ( d, J=6.10 Hz, 2 H) 3.76 (d, J=11.60 Hz, 2 H) 3.31 (t, J=7.10 Hz, 1 H) 2.81 - 2.98 (m, 3 H) 2.70 (s, 2 H) 2.52 - 2.63 (m, 2 H) 2.00 - 2.08 (m, 1 H) 1.53 - 1.62 (m, 2 H).

Example 17:

3-(1-(3-(5-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine- 4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (50 mg, 0.182 mmol) and (3-(6-oxo-) in NMP (3 ml) 1-(3-(5-(piperidin-4-ylmethoxy)pyrimidin-2-yl)benzyl)-1,6-dihydropyridazin-3-yl)benzonitrile (120 mg, 0.182 mmol) To the solution, ethylbis(propan-2-yl)amine (3.0 equivalents) was added to the reaction mixture.The reaction was stirred for 24 hours at 130° C. The residue was purified by MeOH/DCM (0~20%) MPLC, The title compound (80 mg) was obtained.

MS (ESI, m/z): [M+H] ⁺ = 735.75

¹ H NMR (500 MHz, DMSO-d 6 ) d ppm 11.02 (s, 1 H) 8.59 (s, 2 H) 8.31 (d, J=1.83 Hz, 2 H) 8.13 - 8.22 (m, 2 H) 8.11 (d, J=9.77 Hz, 1 H) 7.86 (d, J=7.78 Hz, 1 H) 7.65 (t, J=7.86 Hz, 1 H) 7.60 (d, J=8.54 Hz, 1 H) 7.37 - 7.45 (m, 2 H) 7.28 (s, 1 H) 7.19 (dd, J=8.70, 1.98 Hz, 1 H) 7.09 (d, J=9.61 Hz, 1 H) 5.38 (s, 2 H) 5.00 (dd, J=12.82, 5.34 Hz, 1 H) 3.98 - 4.07 (m, 4 H) 3.19 - 3.24 (m, 1 H) 2.96 (t, J=11.90 Hz, 2 H) 2.77 - 2.89 (m, 1 H) 2.63 (s, 2 H) 2.45 - 2.56 (m, 2 H) 1.90 - 2.01 (m, 1 H) 1.24 - 1.38 (m, 2 H).

Experiment example :

Experimental Example 1: Cell line culture and compound treatment

Human gastric cancer cell lines MKN45 and SNU638 overexpressing c-MET were cultured in RPMI medium supplemented with 10% fetal bovine serum, penicillin (100U/ml), and streptomycin (100mg/ml), and c-MET exon 14 Deletion (exon 14 skipping) mutant human gastric cancer cell line Hs746T was cultured in DMEM medium. The human lung cancer cell line EBC-1 overexpressing c-MET was cultured in MEM medium supplemented with 10% fetal bovine serum, penicillin (100U/ml), and streptomycin (100mg/ml). The normal cell line WI-38 derived from human lung tissue was cultured in EMEM medium supplemented with 10% fetal bovine serum, penicillin (100U/ml), and streptomycin (100mg/ml).

The compound synthesized according to the present invention was prepared as a 10 mM raw material solution dissolved in DMSO (dimethyl sulfoxide), and was sequentially diluted and treated.

Experimental Example 2: Evaluation of c-MET protein degradation ability of compounds of the present invention

In order to evaluate the resolution of the compound of the present invention for c-MET expressed in cells, cells were distributed at a concentration of 2 × 10 ⁵ cells/well in a 12 well plate, and the gastric cancer cell line samples were incubated for 24 hours. and lung cancer cell lines were treated for 48 hours.

To perform a western blotting assay to confirm protein expression, cells treated with the compound were treated with 40 μL of RIPA buffer containing a protease inhibitor for 30 minutes, then placed on ice. Cell lysates were collected from above using a scraper and centrifuged at 4°C at 15,000 rpm for 30 minutes to obtain a supernatant containing protein.

Afterwards, protein expression was measured using the Simple Western Automated Western Blot System Abby instrument using 3 μg of protein per sample. Proteins denatured by SDS and heat were separated by molecular weight and attached to a capillary, then reacted with the primary antibody, anti-c-MET or anti-GAPDH antibody, and then with HRP-conjugated secondary antibody. After reaction, protein expression signal was analyzed using compass software.

Protein expression was normalized based on the expression value of GAPDH, and the amount of protein expression in each sample was measured as a percentage (%) based on 100% samples treated with 0.1% DMSO vehicle, and the amount of protein expression was measured according to compound treatment. DC ₅₀ (the concentration of the compound at which 50% inhibition of protein expression is achieved) was calculated from the change in protein expression level.

The results are shown in Tables 1 to 4 and Figures 1 and 2.

[Table 1] DC ₅₀ and IC ₅₀ of the compounds of the present invention in MKN45 cell line

[Table 2] DC ₅₀ and IC ₅₀ of the compounds of the present invention in SNU638 cell line

[Table 3] DC ₅₀ and IC ₅₀ of the compounds of the present invention in HS746T cell line

[Table 4] DC ₅₀ and IC ₅₀ of the compounds of the present invention in EBC-1 cell line

As a result of the experiment, the majority of compounds showed excellent resolution in the gastric cancer cell line MKN45. In particular, as shown in Tables 1 to 4 and Figures 1 and 2, the compounds of the present invention were effective for all gastric cancer cell lines and lung cancer cell lines used in the experiment. c-MET protein was effectively degraded.

Experimental Example 3: Evaluation of cancer cell anti-proliferative activity by the compound of the present invention

To observe the inhibitory activity on cell growth, cells were distributed in a 96 well plate at a concentration of 5,000 cells/well and then treated with the compounds of the present invention prepared at various concentrations for 72 hours. The survival rate of cells was determined by adding 10 ml per well of WST-8 (water soluble tetrazolium salt) sample, which produces formazan by reacting with dehydrogenase present in the electron transport chain of mitochondria. The culture was incubated for 2 hours, and the absorbance was measured at 450 nm, and the obtained value was calculated using GraphicPad Prism 8.0 software. The IC ₅₀ value (concentration of compound that achieves 50% inhibition of cell proliferation) is the average value of the measurement results. The results are shown in Tables 1 to 4 and Figures 3 and 4.

As shown in the experimental results, the compounds of the present invention showed an effect of inhibiting the proliferation of cancer cells, and in particular, the compounds of Examples 1, 2, and 10, which effectively decompose c-MET, inhibited gastric cancer cell lines at low concentrations of less than 5 nM. The proliferation of MKN45 was effectively inhibited (see Table 1 and Figure 3). In addition, the compounds of Examples 1, 2, and 10 effectively inhibited the proliferation of cancer cells in other gastric cancer cell lines, SNU638 and Hs746T, and lung cancer cell line EBC-1, at a concentration of less than 100 nM.

Experimental Example 4: Evaluation of normal cell toxicity by the compound of the present invention

In order to observe whether the growth of normal cells is inhibited by treatment with the compound of the present invention, the normal cell line WI-38 was dispensed at a concentration of 5,000 cells/well in a 96 well plate, and then the compound of the present invention prepared at various concentrations was dispensed. was treated for 72 hours. The survival rate of cells was measured by adding 10 ml of WST-8 (water soluble tetrazolium salt) sample per well, which reacts with dehydrogenase present in the electron transport chain in mitochondria to produce formazan, and culturing for 2 hours. , the absorbance was measured at 450 nm and the obtained value was calculated using GraphicPad Prism 8.0 software. The IC ₅₀ value (concentration of compound that achieves 50% inhibition of cell proliferation) is the average value of the measurement results. The results are shown in Table 5 and Figure 5.

[Table 5] IC ₅₀ of the compounds of the present invention in WI-38 cell line

As a result of the experiment, it was confirmed that the compounds of the present invention did not affect the proliferation of normal cell lines.

Experimental Example 5: Evaluation of c-MET signaling inhibition by compounds of the present invention

In order to evaluate the effect of the compound of the present invention on inhibiting cancer cell growth signals by decomposing c-MET, the gastric cancer cell line SNU638 was dispensed at a concentration of 5 × 10 ⁵ cells/well in a 6 well plate, and then the compound of the present invention prepared at various concentrations was dispensed. was treated with the compound for 24 hours.

In addition, in order to confirm how the inhibitory effect of cancer cell growth signal transmission is maintained by c-MET protein degradation induced by the compound of the present invention, samples were treated at a concentration of 30 nM, and cells were obtained over time.

To perform a western blotting assay to confirm protein expression, cells treated with the compound were treated with 70 μL of RIPA buffer containing protease inhibitors for 30 minutes, then cell lysates were collected using a scraper on ice, and The supernatant containing protein was obtained by centrifugation at 15,000 rpm for 30 minutes.

Afterwards, SDS-PAGE and membrane transfer of proteins were performed using 10 μg of protein per sample, and anti-phospho-c-MET, anti-phospho-ERK, anti-phospho-AKT, anti- c-MET, anti-ERK, anti-AKT or anti-GAPDH antibody was reacted at 4°C for 16 hours, secondary antibody was reacted at room temperature for 1 hour, and SuperSignal™ West Pico PLUS chemiluminescence substrate ) and an image analyzer were used to confirm protein expression. The results are shown in Figures 6 and 7.

As shown in the results of Figure 6, the compounds of the present invention generally inhibited the activation of ERK and AKT signals in cancer cells induced by c-MET in proportion to the treatment concentration. In addition, as shown in the results of Figure 7, it was confirmed that inhibition of ERK and AKT signaling in cancer cells was maintained for a longer period of time compared to cell lines treated with Tepotinib, a small molecular compound inhibitor.

Experimental Example 6: Evaluation of anticancer activity of the compound of the present invention in an animal model transplanted with gastric cancer cells

To evaluate the inhibitory effect of the compound of the present invention on cancer cell growth by decomposing c-MET in an animal model, 1 × 10 ⁷ gastric cancer cell line SNU638 was transplanted into immunodeficient mice to form tumors of a certain size.

In order to evaluate the anti-cancer activity of the compound of the present invention, gastric mice were randomly divided into groups (5 mice distributed to each group), and the carrier and the compound of the present invention were administered intraperitoneally (FIG. 7) and orally (FIG. 8). The mice were treated every day of the week, and the body weight and tumor size of the mice were measured every three days.

After the experiment was completed, the tumor was extracted and its weight was measured. In order to observe changes in the expression of c-MET protein in the tumor, the tumor was crushed to obtain the protein, which was then confirmed using western blotting assay. The results are shown in Figures 8 and 9, respectively.

As can be seen from the experimental results, little change in the body weight of mice was observed during the treatment period of the compound, and when the compound of the present invention was administered intraperitoneally or orally, it effectively inhibited the growth of human gastric cancer cell lines transplanted into mice. It induced tumor death.

In addition, as can be seen from Figure 8, when the protein expression of the tumor extracted after the end of the experiment was observed, it was confirmed that the expression of c-MET in the group treated with the compound of the present invention was significantly lower than that in the control group.

The compound according to the present invention can be usefully used as an anticancer agent.

Claims (9)

A compound of formula 1 below or a pharmaceutically acceptable salt thereof:
[Formula 1]

In the above formula,
Z is the targeting protein ligand,
Q ₁ to Q ₄ are each independently CZ, CH, C-Hal or C≡N (where Hal is halogen), provided that any one of Q ₁ to Q ₄ is CZ,
R ¹ is hydrogen, halogen, nitro, amino, straight-chain, branched-chain or cyclic alkyl of C ₁ to C ₆ , or straight-chain, branched-chain or cyclic alkyl of C ₁ to C ₆ substituted by halogen. The compound according to claim 1, wherein the targeting protein ligand is a c-MET inhibitor. The method of claim 1, wherein the targeting protein ligand is a compound selected from the group consisting of the following compounds:
The compound of claim 1, wherein R ¹ is hydrogen, halogen, straight-chain, branched-chain or cyclic alkyl of C ₁ to C ₄ , or straight-chain, branched-chain or cyclic alkyl of C ₁ to C ₄ substituted with halogen. . A compound of formula 2 below or a pharmaceutically acceptable salt thereof:
[Formula 2]

In the above formula,
Y is -C=O or -CH ₂ ,
Z is the targeting protein ligand,
Q ₁ to Q ₄ are each independently CZ, CH, C-Hal or C≡N (where Hal is halogen), provided that any one of Q ₁ to Q ₄ is CZ. The compound according to claim 5, wherein the targeting protein ligand is a c-MET inhibitor. The method of claim 5, wherein the targeting protein ligand is a compound selected from the group consisting of the following compounds:
A compound selected from the group consisting of the following compounds or a pharmaceutically acceptable salt thereof:

A compound selected from the group consisting of the following compounds or a pharmaceutically acceptable salt thereof:
3-(6-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]tria Zolo[4,5-b]pyrazin-1-yl)methyl)morpholino)pyrimidin-5-yl)benzyl)piperazin-1-yl)-1-oxophthalazin-2(1H)-yl ) piperidine-2,6-dione
3-(4-methyl-6-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2 ,3] triazolo [4,5-b] pyrazin-1-yl) methyl) morpholino) pyrimidin-5-yl) benzyl) piperazin-1-yl) -1-oxophthalazine-2 ( 1H)-yl)piperidine-2,6-dione
3-(7-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]tria Zolo[4,5-b]pyrazin-1-yl)methyl)morpholino)pyrimidin-5-yl)benzyl)piperazin-1-yl)-1-oxophthalazin-2(1H)-yl ) piperidine-2,6-dione
3-(8-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]tria Zolo[4,5-b]pyrazin-1-yl)methyl)morpholino)pyrimidin-5-yl)benzyl)piperazin-1-yl)-1-oxophthalazin-2(1H)-yl ) piperidine-2,6-dione
3-(4-methyl-8-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2 ,3] triazolo [4,5-b] pyrazin-1-yl) methyl) morpholino) pyrimidin-5-yl) benzyl) piperazin-1-yl) -1-oxophthalazine-2 ( 1H)-yl)piperidine-2,6-dione
3-(6-fluoro-7-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazol-4-yl)-1H-[1, 2,3]triazolo[4,5-b]pyrazin-1-yl)methyl)morpholino)pyrimidin-5-yl)benzyl)piperazin-1-yl)-1-oxophthalazine-2 (1H)-yl)piperidine-2,6-dione
2-(2,6-dioxopiperidin-3-yl)-4-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazole-4 -yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl)methyl)morpholino)pyrimidin-5-yl)benzyl)piperazin-1-yl) Isoindoline-1,3-dione
2-(2,6-dioxopiperidin-3-yl)-5-(4-(4-(2-((S)-2-((5-(1-methyl-1H-pyrazole-4 -yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl)methyl)morpholino)pyrimidin-5-yl)benzyl)piperazin-1-yl) Isoindoline-1,3-dione
3-(1-(3-(5-((1-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl )piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile
3-(1-(3-(5-((1-(2-(2,6-dioxopiperidin-3-yl)-4-methyl-1-oxo-1,2-dihydrophthalazine -6-yl)piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile
3-(1-(3-(5-((1-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-5-yl )piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile
3-(1-(3-(5-((1-(3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrophthalazin-6-yl )piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile
3-(1-(3-(5-((1-(3-(2,6-dioxopiperidin-3-yl)-4-oxo-3,4-dihydrophthalazin-5-yl )piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile
3-(1-(3-(5-((1-(3-(2,6-dioxopiperidin-3-yl)-1-methyl-4-oxo-3,4-dihydrophthalazine -6-yl)piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile
3-(1-(3-(5-((1-(2-(2,6-dioxopiperidin-3-yl)-7-fluoro-1-oxo-1,2-dihydrophthala zin-6-yl)piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile
3-(1-(3-(5-((1-(3-(2,6-dioxopiperidin-3-yl)-7-fluoro-4-oxo-3,4-dihydrophthala zin-6-yl)piperidin-4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile
3-(1-(3-(5-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperidine- 4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile
3-(1-(3-(5-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine- 4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile