KR102489160B1 - Derivatives of Piperidinedione - Google Patents

Abstract

A novel piperidinedione derivative compound is disclosed.

Description

Piperidinedione derivative {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, an abbreviation of PROteolysis TArgeting Chimeras, is a technology that induces the degradation of a target protein (POI: protein of interest). Instead of the term protac, 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 FIG. 1). The target protein ligand is a structure that can bind to a target protein associated with a disease, and the E3 enzyme ligand is a structure that can bind to an E3 enzyme.

E3 enzyme (E3 ligase) can attach a label called ubiquitin to a target protein, and the target protein labeled with such ubiquitin is 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 Protac compound, thus 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 achieve a cancer treatment effect by inducing degradation of the target protein.

WO 2010/078897 A1;

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

The present invention is to provide a compound that can be used for TPD by increasing the decomposition effect of a target protein (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 a target protein ligand;

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

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

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

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

In addition, the present invention relates to a compound of Formula 2 or a pharmaceutically acceptable salt thereof:

[Formula 2]

In the above formula,

Y is -C=0 or -CH ₂ ;

Z is a target protein ligand,

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

In the compound of the present invention, a c-MET inhibitor compound 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:

In addition, 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-oxophthalazin-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-oxophthalazin-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-oxophthalazin-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)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-5-yl)piperidin- 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 comprising a target protein ligand and an E3 enzyme ligand, and since the target protein ligand and E3 enzyme ligand are directly covalently linked to each other without using a linker, it is useful as a compound for target protein degradation. can be used 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 protein. It is possible to decompose

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

[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, C ₁ to C ₆ straight-chain, branched or cyclic alkyl, or C ₁ to C ₆ straight-chain, branched or cyclic alkyl substituted by halogen (preferably , Wherein R ¹ is hydrogen, halogen, C ₁ to C ₄ straight-chain, branched or cyclic alkyl, or halogen-substituted C ₁ to C ₄ straight-chain, branched or cyclic alkyl).

[Formula 4]

In the above formula,

Y is -C=0 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 target protein ligand, nuclear receptor (e.g., ER, AR, RAR, etc.), protein kinase (e.g., Akt, c-Abl, BTK, anaplastic lymphoma kinase [ALK], CDK9, c-Met, RIPK2, DAPK1, PSD-95, etc.), proteins involved in transcriptional regulation (eg, BRD4, Sirt2, HDAC6, TRIM24, IKZH1/3, Smad3, etc.), regulatory proteins (e.g. CRABP-I/II, TACC3, AHR, FKBP12, ERRα, SHP2, X-protein, PTPN11, etc.), neurodegenerative related proteins (e.g. Huntingtin, Tau, a-synuclein, PSD-95 etc.), cellular metabolic enzymes (eg MetAP-2, DHODH), fusion proteins (eg Alk-fusion, BCR-Abl, Brd-Nut, Ret-fusion, Halo Tags, etc.), mutant proteins (eg Alk-fusion, BCR-Abl, Brd-Nut, Ret-fusion, Halo Tags, etc.) For example, compounds that target proteins such as Braf mutant, EGFR mutant and deletion, Kras mutant, TP53 mutant and splicing variant, etc.), abnormal modified protein (eg EGFRdel19, P53 splicing variant, fusion proteins, etc.) can be used. However, preferably, a compound targeting the c-MET protein may be used as the target protein ligand. Therefore, the compound according to the present invention can be usefully used for the treatment or prevention of diseases related to c-MET protein.

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

In the present invention, the right side of the target protein ligand may covalently bind to any one carbon atom of Q1 to Q4 in the chemical compound of 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 during the covalent bonding reaction at the site where the target protein ligand and the E3 enzyme ligand are covalently bonded to each other.

Since the compound of the present invention can bind to and degrade a target protein (eg, c-MET protein), the protac compound can be usefully used for the treatment or prevention of diseases related to the target protein. For example, the compound of the present invention has an excellent anticancer effect, and further induces the degradation of c-MET protein to exhibit a therapeutic effect on diseases related to c-MET.

1 schematically shows a protac compound consisting of a target protein ligand, a linker, and an E3 enzyme ligand.
Figure 2 shows the experimental results confirming the proteolytic effect of the compound of the present invention in the MKN45 cell line.
Figure 3 shows the experimental results confirming the proteolytic effect of the compound of the present invention in SNU638, Hs746T, EBC-1 cell lines.
Figure 4 shows the experimental results confirming the MKN45 cancer cell proliferation inhibitory effect of the compound of the present invention.
Figure 5 shows the experimental results confirming the inhibition of SNU638, Hs746T, EBC-1 cancer cell proliferation of the compound of the present invention.
Figure 6 shows the experimental results confirming whether or not the compound of the present invention has an inhibitory effect on normal cell proliferation.
Figure 7 shows the experimental results confirming the cell signaling inhibitory effect of each concentration of the compound of the present invention.
Figure 8 shows the experimental results confirming the cell signaling inhibitory effect over time of the compound of the present invention in SNU638 cells.
Figure 9 shows the experimental results confirming the anticancer effect by intraperitoneal administration of the compound of the present invention in mice transplanted with SNU638 tumor cells.
Figure 10 shows the experimental results confirming the anticancer effect by oral administration of the compound of the present invention in mice transplanted with SNU638 tumor cells.

Hereinafter, the present invention will be described in detail based on examples to aid understanding of the present invention. However, the following examples are merely illustrative of the contents of the present invention, and the spirit or scope of the present invention is not limited in any sense by the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.

Example A: oxophthalazinyl piperidinedione Preparation of derivative compounds

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

[Scheme 1]

[Scheme 2]

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

Hereinafter, specific examples of the compound of Formula 1 are prepared through Examples.

Example A1:

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

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

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

1-Bromopyrrolidine-2,5-dione (24.7g, 139mmol) was added to a solution of methyl 2-fluoro-6-methylbenzoate (21.2g, 126 mmol) in ClCH ₂ CH ₂ Cl (250ml) at room temperature. and benzoylbenzenecarboperoxoate (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 completion point of the reaction. 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, yield 112%) MS (ESI, m/z): [M+1] ⁺ = [245.9].

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

At room temperature, to a solution of methyl 2-(bromomethyl)-6-fluorobenzoate (35 g, 142 mmol) in DCM (280 ml) 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 (200ml), 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, yield 45%) MS (ESI, m/z): [M+1] ⁺ = [183.1].

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

At room temperature, to a solution of methyl 2-fluoro-6-formylbenzoate (11.5 g, 63.2 mmol) in THF (82 ml) was added lithium(+1) hydroxide (7.57 g, 180 mmol) aqueous solution (41 ml). . After stirring for 4 hours, the reactant was concentrated under reduced pressure to dry THF. 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 layers were dried over 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, to a solution of 2-fluoro-6-formylbenzoic acid (10.4g, 61.9mmol) in methanol (120ml) was added NH ₂ NH ₂ monohydrate (3.72mg, 61.9mmol) and stirred at room temperature for 16 hours. . The white precipitate was filtered off 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

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

Example A2:

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

At room temperature, to a solution of 3-fluoro-2-methylbenzoic acid (10 g, 64.9 mmol) in methanol (60 ml) was added sulfuric acid (2 ml). Heated to 80 ^° C and stirred overnight. After cooling to room temperature, the reaction was evaporated and extracted with NaHCO ₃ and ethyl acetate (EA). Dried over MgSO ₄ . The crude product was used in the next reaction without further purification. (10.1 g, yield 92%) 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) was added to 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 benzoylbenzenecarboperoxoate (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 completion point of the reaction. 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, yield 71%) MS (ESI, m/z): [M+1] ⁺ = [245.9].

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

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), 4 Å molecular sieves at room temperature. The reaction was stirred at room temperature for 4 hours. The DCM layer was washed with water (200ml), dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by column chromatography to give the title compound. (5.5 g, 75%) MS (ESI, m/z): [M+1] ⁺ = [183.1].

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

At room temperature, a solution of methyl 3-fluoro-2-formylbenzoate (2.5g, 13.7mmol) in THF (68ml) in aqueous solution of lithium(+1) hydroxide monohydrate (2.88g, 68.6mmol) (41ml) was added. After stirring for 4 hours, the reactant was concentrated under reduced pressure to dry THF. 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 layers were dried over 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

To a solution of 3-fluoro-2-formylbenzoic acid (2.5g, 14.9mmol) in THF:water=1:1 (70ml) at room temperature, NH ₂ NH ₂ monohydrate (1.22mg, 16.4mmol) was added, and 16 Stir for an hour. The mixture was acidified to pH 4 and the solid was filtered with hexanes. Drying in vacuo 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 (526mg, 2.74mmol) was added to the solution and stirred at 80°C for 6 hours. Upon completion of the reaction, the reactant was cooled to room temperature, poured into water (100ml), the pH was adjusted to 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 in vacuo to give the title compound. MS (ESI, m/z): [M+1] ⁺ = [276.1].

Example A3

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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

To a solution of methyl 4-fluoro-2-methylbenzoate (20 g, 119 mmol) in ClCH ₂ CH ₂ Cl (100 ml) at room temperature, 1-bromopyrrolidine-2,5-dione (31.8 g, 178 mmol) and benzoyl Benzenecarboperoxoate (1.92g, 5.95mmol) was added sequentially. The reaction was heated to reflux for 3 hours. The point at which the red color disappears is the completion point of the reaction. 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

To a solution of methyl 2-(bromomethyl)-4-fluorobenzoate (22g, 89mmol) in DCM (100ml) at room temperature, NMO (15.6mg, 134mmol) and 4A molecular sieves were added sequentially. The reaction was stirred at room temperature for 2 hours. The molecular sieve was filtered and washed with DCM (50ml). The DCM layer was washed with water (200ml), 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. (12 g, 73.98%) MS (ESI, m/z): [M+1] ⁺ = [183.2].

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

At room temperature, to a solution of methyl 4-fluoro-6-formylbenzoate (12 g, 65.9 mmol) in THF (50 ml) was added an aqueous solution of lithium(+1) hydroxide (13.8 g, 329 mmol) (50 ml). After stirring for 2 hours, the reaction mixture was concentrated under reduced pressure to dry THF. 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 over MgSO ₄ and concentrated under reduced pressure to obtain white crystals (10 g, 90.3%). MS (ESI, m/z): [M+1] ⁺ = [169.2].

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

At room temperature, to a solution of 4-fluoro-2-formylbenzoic acid (6.78 g, 40.3 mmol) in methanol (50 ml) was added NH ₂ NH ₂ monohydrate (2.02 g, 40.3 mmol). 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 over 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-methylpropan-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. 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 over MgSO ₄ and concentrated under reduced pressure. The residue was purified by chromatography to give the title compound as white crystals (110 mg, 65.5%). MS (ESI, m/z): [M+1] ⁺ = [276.6].

Example A4

의 제조compound

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

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

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

To a solution of methyl 5-fluoro-2-methylbenzoate (20 g, 119 mmol) in ClCH ₂ CH ₂ Cl (100 ml) at room temperature, 1-bromopyrrolidine-2,5-dione (31.8 g, 178 mmol) and benzoyl Benzenecarboperoxoate (1.92g, 5.95mmol) was added sequentially. The reaction was heated to reflux for 3 hours. The point at which the red color disappears is the completion point of the reaction. 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

To a solution of methyl 2-(bromomethyl)-5-fluorobenzoate (29 g, 117 mmol) in DCM (100 ml) at room temperature, NMO (20.6 mg, 176 mmol) and 4A molecular sieves were added sequentially. The reaction was stirred at room temperature for 2 hours. The molecular sieve was filtered and washed with DCM (50ml). The DCM layer was washed with water (200ml), 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) Preparation of 5-fluoro-2-formylbenzoic acid

At room temperature, to a solution of methyl 5-fluoro-2-formylbenzoate (15 g, 82.3 mmol) in THF (50 ml) was added lithium(+1) hydroxide (17.3 g, 412 mmol) aqueous solution (50 ml). After stirring for 2 hours, the reaction mixture was concentrated under reduced pressure to dry THF. 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 over 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, to a solution of 5-fluoro-2-formylbenzoic acid (8.16g, 48.5mmol) in methanol (50ml) was added NH ₂ NH ₂ monohydrate (3.74g, 74.8mmol). 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 over 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-methylpropan-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. added After stirring for 30 minutes, 3-bromopiperidine-2,6-dione (1.05mg, 5.48mmol) 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 over MgSO ₄ and concentrated under reduced pressure. The residue was purified by column chromatography to give the title compound as white crystals (300 mg, 35.78%). MS (ESI, m/z): [M+1] ⁺ = [276.6].

Example A5

의 제조 (반응식 2에 준하여 제조함)compound

Preparation of (prepared according to Scheme 2)

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

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) was added tetrakis(triphenylphosphine). )-palladium(0) (557mg, 0.48mmol) was added and stirred at 100°C for 16 hours. After cooling, 5 ml of 1N-HCl was added and stirred for 1 hour. The organic layer was dried over MgSO ₄ and concentrated in the dark. The residue was purified by silica column chromatography to give the title compound as a yellow oil. (820 mg, yield 87%) 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 (810mg, 4.13mmol) in THF (20ml) and water (10ml) was added LiOH (494mg, 20.6mmol) and stirred at room temperature for 20 hours. The solution was acidified with 1N-HCl to a pH of about 3. After adding 100ml EA, 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 was added 1M-Lithium diisopropylamide (3.6 ml, 3.6 mmol), Stir for 20 minutes. 3-brobopiperidine-2,6-dione (863 mg, 4.5 mmol) was added to the solution, and the mixture was stirred at 80°C for 2 hours. The reactant was concentrated under reduced pressure and dried. After adding water (10 ml) and stirring for 1 hour, the pH was adjusted to 4 by acidification with 1N-HCl. 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 of ClCH ₂ CH ₂ Cl was added AIBN (401 mg, 2.44 mmol), 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 a residue to which 50 ml water was added. The mixture was refluxed with stirring for 4 hours, the mixture was cooled, the product was filtered, neutral washed with water and dried to give pale off-white crystals. (4.85 g, 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) 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 give white crystals. (1.31 g, 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 1.0 M LDA (7.33 mL) dropwise at 0 °C. After stirring for 30 minutes, 3-bromopiperidine-2,6-dione was added portionwise to the reaction. The reaction was heated to 80 °C and stirred for 2 hours. Upon completion of the reaction, the reactant was cooled to room temperature, poured into water (100 ml), the pH was adjusted to 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 give 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 eq) were added. The reaction was stirred for 2 hours at room temperature. 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

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 give 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 It was 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, 46.6 mmol) was added triethylamine (3.0 equiv.). 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 give 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) To the solution was added isoamyl nitrite (3.8 g, 32.5 mmol). 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 (7.5 g).

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 Synthesis of -1-yl]methyl}morpholine-4-carboxylate

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- To a solution of dioxaborolan-2-yl)-1H-pyrazole (4.7 g, 22.5 mmol) and cesium carbonate (18.4 g, 56.4 mmol) was added Pd(dppf)Cl2 (0.7 g, 0.95 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 give 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) was added TFA (100 ml). The reaction was stirred for 2 hours at room temperature. 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, Synthesis of 3]triazolo[4,5-b]pyrazin-1-yl]methyl}morpholine

(2S)-2-{[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazine in EtOH (300 ml) To a solution of -1-yl]methyl}morpholine (4.8 g, 16 mmol) and 5-bromo-2-chloropyrimidine (4.64 g, 24 mmol) ethylbis(propan-2-yl)amine (3.0 eq.) 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- Synthesis of b]pyrazin-1-yl]methyl}morpholin-4-yl]pyrimidin-5-yl}benzaldehyde

(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, 4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde (4.2 g, 18 mmol), Pd(dppf)Cl2 in a solution of 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) was added sodium triacetoxyborohydride (3.9 g, 18.7 mmol) and AcOH (1 ml). 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 (6.7 g).

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- yl]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) solution was added TFA (50 ml). The reaction was stirred for 2 hours at room temperature. 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 (6.7 g).

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) (3ml) , 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 -yl)amine (3.0 eq) 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 give 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-oxophthalazin-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 )-2-{[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl]methyl}- To a solution of 4-(5-{4-[(piperazin-1-yl)methyl]phenyl}pyrimidin-2-yl)morpholine (100 mg, 0.182 mmol) was added 3.0 eq) 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 to give 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 (3ml) {[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 (100mg, 0.182 mmol) reacted with ethylbis(propan-2-yl)amine (3.0 eq.) 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 give 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 (3ml) {[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 (100mg, 0.182 mmol) reacted with ethylbis(propan-2-yl)amine (3.0 eq.) 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 give 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-oxophthalazin-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 )-2-{[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl]methyl}- To a solution of 4-(5-{4-[(piperazin-1-yl)methyl]phenyl}pyrimidin-2-yl)morpholine (100 mg, 0.182 mmol) was added 3.0 eq) 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 to give 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-oxophthalazin-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)- 2-{[5-(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl]methyl}-4- Ethylbis(propan-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 to give 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 (50mg, 0.182 mmol) and (2S)-2-{[5 in NMP (3ml) -(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl]methyl}-4-(5-{4 To a solution of -[(piperazin-1-yl)methyl]phenyl}pyrimidin-2-yl)morpholine (100 mg, 0.182 mmol) was added ethylbis(propan-2-yl)amine (3.0 equiv) to the reaction mixture. added The reaction was stirred at 130° C. for 24 hours. The residue was purified by MeOH/DCM (0-20%) MPLC to give 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 (50mg, 0.182 mmol) and (2S)-2-{[5 in NMP (3ml) -(1-methyl-1H-pyrazol-4-yl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl]methyl}-4-(5-{4 To a solution of -[(piperazin-1-yl)methyl]phenyl}pyrimidin-2-yl)morpholine (100 mg, 0.182 mmol) was added ethylbis(propan-2-yl)amine (3.0 equiv) to the reaction mixture. added The reaction was stirred at 130° C. for 24 hours. The residue was purified by MeOH/DCM (0-20%) MPLC to give 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 -(6-oxo-1-(3-(5-(piperidin-4-ylmethoxy)pyrimidin-2-yl)benzyl)-1,6-dihydropyridazin-3-yl)benzonitrile ( 120mg, 0.182mmol) ethylbis(propan-2-yl)amine (3.0 equiv.) was added to the reaction mixture.The reaction was stirred for 24 hours at 130°C. The residue was obtained in MeOH/DCM (0-20% ) 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 -(6-oxo-1-(3-(5-(piperidin-4-ylmethoxy)pyrimidin-2-yl)benzyl)-1,6-dihydropyridazin-3-yl)benzonitrile ( 120mg, 0.182mmol) ethylbis(propan-2-yl)amine (3.0 equiv.) was added to the reaction mixture.The reaction was stirred for 24 hours at 130°C. The residue was obtained in MeOH/DCM (0-20% ) 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- Oxo-1-(3-(5-(piperidin-4-ylmethoxy)pyrimidin-2-yl)benzyl)-1,6-dihydropyridazin-3-yl)benzonitrile (120 mg, 0.182 mmol) solution, ethylbis(propan-2-yl)amine (3.0 equivalent) 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 This gave the title compound (90 mg).

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- Oxo-1-(3-(5-(piperidin-4-ylmethoxy)pyrimidin-2-yl)benzyl)-1,6-dihydropyridazin-3-yl)benzonitrile (120 mg, 0.182 mmol) solution, ethylbis(propan-2-yl)amine (3.0 equivalent) 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 This gave the title compound (40 mg).

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, 2H).

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- Oxo-1-(3-(5-(piperidin-4-ylmethoxy)pyrimidin-2-yl)benzyl)-1,6-dihydropyridazin-3-yl)benzonitrile (120 mg, 0.182 mmol) solution, ethylbis(propan-2-yl)amine (3.0 equivalent) 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 This gave the title compound (40 mg).

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 -(6-oxo-1-(3-(5-(piperidin-4-ylmethoxy)pyrimidin-2-yl)benzyl)-1,6-dihydropyridazin-3-yl)benzonitrile ( 120mg, 0.182mmol) ethylbis(propan-2-yl)amine (3.0 equiv.) was added to the reaction mixture.The reaction was stirred for 24 hours at 130°C. The residue was obtained in MeOH/DCM (0-20% ) 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) solution of ethylbis(propan-2-yl)amine (3.0 equivalent) was added to the reaction mixture.The reaction was stirred for 24 hours at 130° C. 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)piperidin- 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- 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 equivalent) was added to the reaction mixture to the solution.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)piperidin- 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- 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 equivalent) was added to the reaction mixture to the solution.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).

Experimental example :

Experimental Example 1: Cell line culture and compound treatment

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

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

Experimental Example 2: Evaluation of c-MET protein degrading ability of the compound 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 dispensed at a concentration of 2 × 10 ⁵ cells/well in a 12-well plate, and gastric cancer cell lines were sampled for 24 hours. and lung cancer cell lines were treated for 48 hours.

To perform a western blotting assay to confirm protein expression, compound-treated cells were treated with 40 μL of RIPA buffer to which protease inhibitors were added for 30 minutes, and then incubated on ice. The cell lysate was collected using a scraper from above, and centrifuged at 4° C. and 15,000 rpm for 30 minutes to obtain a protein-containing supernatant.

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

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

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

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

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

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 compounds of the present invention in EBC-1 cell line

As a result of the experiment, most of the compounds in the gastric cancer cell line MKN45 showed excellent resolution. In particular, as shown in the results of Tables 1 to 4 and FIGS. 1 and 2, the compounds of the present invention were used 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 antiproliferative activity by the compound of the present invention

In order to observe the inhibitory activity on cell growth, the cells were dispensed in a 96 well plate at a concentration of 5,000 cells/well, and then treated with the compound of the present invention prepared at various concentrations for 72 hours. Cell viability was measured by adding 10 ml of WST-8 (water soluble tetrazolium salt) sample per well, which reacts with dehydrogenase present in the mitochondrial electron transport system to produce formazan, After incubation for 2 hours, absorbance was measured at 450 nm and the obtained value was calculated using GraphicPad Prism 8.0 software. The IC ₅₀ value (the concentration of the 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 FIGS. 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 degrade c-MET, gastric cancer cell lines at low concentrations of less than 5 nM. 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 at a concentration of less than 100 nM in other gastric cancer cell lines, SNU638 and Hs746T, and lung cancer cell line EBC-1.

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 the treatment of 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 treated for 72 hours. Cell viability 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 system to mitochondria to produce formazan, and incubated for 2 hours. , the absorbance was measured at 450 nm and the value obtained was calculated using GraphicPad Prism 8.0 software. The IC ₅₀ value (the concentration of the 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 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 inhibition of c-MET signaling by the compound of the present invention

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

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

To perform a western blotting assay to confirm protein expression, compound-treated cells were treated with 70 μL of RIPA buffer with protease inhibitors added for 30 minutes, then cell lysates were collected using a scraper on ice, and 4 A supernatant containing protein was obtained by centrifugation at 15,000 rpm for 30 minutes.

Thereafter, SDS-PAGE and membrane transfer of proteins were performed using 10 μg of protein for each sample, and anti-phospho-c-MET, anti-phospho-ERK, anti-phospho-AKT, anti- After reacting with c-MET, anti-ERK, anti-AKT or anti-GAPDH antibodies at 4°C for 16 hours, and reacting with secondary antibodies for 1 hour at room temperature, 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 FIG. 6 , the compounds of the present invention 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 FIG. 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 a small molecule inhibitor, tepotinib.

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 cancer cell growth inhibitory effect of the compound of the present invention by c-MET degradation 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 anticancer activity of the compound of the present invention, the stomach mice were randomly divided into groups (distributed 5 mice per group), and the carrier and the compound of the present invention were administered intraperitoneally (FIG. 7) and orally (FIG. 8). The treatment was performed every day for a week, and the body weight and tumor size of the mice were measured every 3 days.

After the experiment was finished, the tumor was removed and weighed. In order to observe the change in c-MET protein expression in the tumor, the protein was obtained by crushing the tumor and confirmed using a western blotting assay. The results are shown in FIGS. 8 and 9 respectively.

As can be seen from the experimental results, almost no 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. Apoptosis of the tumor was induced.

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

The compounds according to the present invention can be usefully used as anticancer agents.

Claims (9)

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

In the above formula,
Q ₁ to Q ₄ are each independently CZ, CH or C-Hal (here, Hal is a halogen), provided that any one of Q ₁ to Q ₄ is CZ;
R ¹ is hydrogen or C ₁ to C ₆ straight-chain, branched-chain or cyclic alkyl;
Z is a target protein ligand, selected from the group consisting of compounds of the following structural formula:

delete delete The compound according to claim 1, wherein R ¹ is hydrogen or C ₁ to C ₄ straight-chain, branched-chain or cyclic alkyl. A compound of Formula 2 or a pharmaceutically acceptable salt thereof:
[Formula 2]

In the above formula,
Y is a carbonyl group,
Q ₁ to Q ₄ are each independently CZ, CH or C-Hal (here, Hal is a halogen), provided that any one of Q ₁ to Q ₄ is CZ;
Z is a target protein ligand, selected from the group consisting of compounds of the following structural formula:

delete delete 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-oxophthalazin-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-oxophthalazin-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-oxophthalazin-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)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-5-yl)piperidin- 4-yl)methoxy)pyrimidin-2-yl)benzyl)-6-oxo-1,6-dihydropyridazin-3-yl)benzonitrile

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