KR20110085036A - Imidazopyrazinone derivatives with apoptosis inducing activity on cells - Google Patents

Abstract

PURPOSE: A pharmaceutical composition containing a novel compound is provided to effectively suppress inflammatory diseases and autoimmune diseases. CONSTITUTION: A pharmaceutical composition for treating cancer, inflammation, autoimmune diseases, and neurodegenerative diseases contains an imidazopyrazinone derivative of chemical formula 1, salt, isomer, hydrate, and solvate thereof. In chemical formula 1, X is oxygen or hydrogen; and R-R8 are independently hydrogen, -COOR9, C1-8 alkyl, C3-8 cycloalkyl, C2-7 heterocycloalkyl, C3-14 aryl, or C2-13 heteroaryl.

Description

IMDAZOPYRAZINONE DERIVATIVES WITH APOPTOSIS INDUCING ACTIVITY ON CELLS}

The present invention relates to novel imidazopyrazinone derivatives having a prophylactic and therapeutic effect on cancer, inflammation, autoimmune diseases, neurodegenerative disorders, etc. caused by overexpression of apoptosis inhibitory proteins and pharmaceutical compositions comprising the same.

Apoptosis or programmed cell death plays a key role in maintaining homeostasis in welfare animals. Such apoptosis maintains life by regulating cell growth and death, but if this system is hampered by a number of factors, it can cause pathological diversity, including cancer, autoimmune diseases, or neurodegenerative disorders. Thompson, CB Science, 267 , 1456-1462 (1995); Hanahan, D. & Weinberg, RA, Cell, 100 , 57-70 (2000)].

In the regulation of apoptosis in the cancer cell development stage, the overexpression of the inhibition of apoptosis protein (IAP) results in the accumulation of IAP protein in the cells, and thus through the apoptosis stage of cancer cells By inhibiting cell proliferation, it inhibits the mechanism of natural cell death in the development and growth and metastasis of cancer cells by various cell death signals (eg DNA damage, chemical agents and stimuli such as ultraviolet light). George George, Biochemistry , 41 , 7344-7349, (2002); Yigong Shi, Nature Rev. Mol. Cell. Bio ., 5 , 897-907, (2004)].

The IAP protein binds to caspase, a cysteine protease involved in cell death, and inhibits cell death. Caspase binds to a portion of more than 70 amino acids, including zinc, called the baculovirus inhibitory repeat (BIR) of the IAP protein. XIAP (human X chromosomeencoded IAP), cIAP-1 (cellular IAP 1) and cIAP-2 (cellular IAP 2) have three side-by-side linked BIR domains at the N-terminus, while other mammalian IAPs have one BIR domain have. XIAP is the most effective caspase inhibitor of the IAP protein family and binds via the BIR-3 domain and the BIR-2 domain, respectively, to the initiation-case phase caspa-9 and the run-case phase cascade 3/7. cIAP-1 and -2 are not yet well known for their role in cell proliferation, but both bind to the TNF-receptor 1 signal complex.

SMAC / Diablo (the second mitochondrial activator of apoptosis / direct IAP-binding protein with low pI) protein, which is released during mitochondrial cell death signaling, is a cascade of IAP proteins from natural IAP proteins. It binds to the same site that binds and modulates the activity of these IAP proteins. In addition, gene amplification of IAP and overexpression of IAP protein are found in many cancer cells.

For this reason, resistance to apoptosis has emerged as an important mechanism for the progression of cancer, and thus targeting cancer cells and other apoptosis mechanisms in tumor cells has been suggested as an effective anticancer drug strategy. In addition, these drugs have no effect on normal cells, and have been reported to have an advantage of minimizing side effects when used alone or in combination therapy, showing activity by acting specifically on cancer cells.

Novartis International Patent Publication Nos. WO2008 / 073305A1, WO2008 / 073306A1, WO2008 / 016893A2, WO2006 / 107963A1, WO2006 / 113376A1, WO2005 / 097791A1, and Genentech International Patent Publications WO2009 / 089502A1, WO2008 / 079735A Multinational pharmaceutical companies, such as Aegera International Patent Publication No. WO2007 / 131366A1 and TetraLogic International Patent Publication No. WO2008 / 014252A2, are also interested in the research.

As a method for inhibiting IAP, studies are being conducted to mimic the structure of SMAC / Diablo, a natural IAP inhibitory protein. These studies have revealed in particular that the core sequence of alanine-valine-proline-isoleucine (Ala-Val-Pro-Ile, AVPI) at the N-terminus is an essential protein that binds to IAP proteins. See Yigong Shi, Nature. structural biology , 8 , 394-401, (2001). These key sequences (AVPI or AVPF) were expected to have pharmacological activity with 120-500 nM activity in vitro activity test, but could not reach drug development due to cell permeability.

Accordingly, the present inventors search for substances having the characteristics of AVPI, a natural IAP inhibitory protein sequence, and cell permeability, and search for the activity of cancer, inflammation, autoimmune diseases and neurodegenerative disorders by using the compounds found. It was. As a result, a parallel synthesis method using combinatorial chemistry and a method of constructing a focused library using fast and highly efficient synthesis by solution phase reaction and searching for activity and potency of AVPI mimic structure In addition, it was possible to develop an imidazopyrazinone derivative having excellent selective efficacy on IAP.

WO2008 / 073305 A1 (NOVATIS AG) 2008.6.19 WO2008 / 073306 A1 (NOVATIS AG) 2008.6.19 WO2008 / 016893 A1 (NOVATIS AG) 2008.2.7 WO2006 / 107963 A2 (NOVATIS AG) 2007.10.12 WO2006 / 113376 A1 (NOVATIS AG) 2006.10.26 WO2005 / 097791 A1 (NOVATIS AG) 2005.10.20 WO2009 / 089502 A1 (GENENTECH.INC) 2009.7.16 WO2008 / 079735 A1 (GENENTECH.INC) 2008.7.3 WO2007 / 131366 A1 (AEGERA THERAPEUTICS INC) 2007.11.22 WO2008 / 014252 A2 (TETRALOGIC PHAM., CO.) 2008.1.31

 Thompson, C. B. Science, 267, 1456-1462 (1995)  Hanahan, D. & Weinberg, R. A., Cell, 100, 57-70 (2000)  George L. M., Biochemistry, 41, 7344-7349, (2002)  Yigong Shi, Nature Rev. Mol. Cell. Bio., 5, 897-907, (2004)  Yigong Shi, Nature structural biology, 8, 394-401, (2001)

Accordingly, it is an object of the present invention to provide novel compounds and pharmaceutical compositions with fewer side effects while selectively and effectively inhibiting cancer cell growth, inflammatory diseases, autoimmune diseases and neurodegenerative disorders by inhibition of cell death.

In accordance with the above object, the present invention provides a compound selected from imidazopyrazinone derivatives having a structure of Formula 1, salts, isomers, hydrates and solvates thereof:

[Formula 1]

In Chemical Formula 1,

X is oxygen or hydrogen;

R ₁ to R ₈ are each independently hydrogen, —COOR ₉ , C _1-8 alkyl, C _3-8 cycloalkyl, C _2-7 heterocycloalkyl, C _3-14 aryl, C _2-13 heteroaryl, or C _1-3 alkyl substituted with one or more C _3-14 aryl or C _2-13 heteroaryl;

The alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are each independently hydrogen, R ₉ , -OH, -OR ₉ , -COR ₉ , -COOH, -COOR ₉ , -CONR ₉ R ₁₀ , -NR ₉ , -NR ₉ CONR ₁₀ R ₁₁ , -CONR ₉ R ₁₀ , -NR ₉ CONR ₁₀ R ₁₁ , -SO ₂ R ₉ , -SO ₂ H and -SOR ₉ ;

R ₉ to R ₁₁ are each independently hydrogen, halogen, amino, nitro, carboxy, cyano, sulfyl, hydroxy, C _1-4 alkyl, C _1-4 alkoxy, C _3-8 cycloalkyl, C _{2 -7} heterocycloalkyl, C _3-14 aryl, or C _2-13 heteroaryl.

According to the above another object, the present invention provides a pharmaceutical composition comprising at least one of the imidazopyrazinone derivative compound, salts, isomers, hydrates and solvates thereof as an active ingredient, and together with the pharmacologically acceptable carrier to provide.

The present invention also provides a compound library comprising at least one of the above imidazopyrazinone derivative compounds, salts, isomers, hydrates and solvates thereof.

The novel imidazopyrazinone derivative compounds and pharmaceutical compositions comprising the same according to the present invention act selectively and effectively on the mechanism of inhibition of apoptosis, which is one of the causes of cancer, inflammation, autoimmune diseases and neurodegenerative disorders. It can be used as an effective prophylactic and therapeutic agent with fewer side effects for diseases such as inflammation, autoimmune diseases and neurodegenerative disorders.

Hereinafter, the present invention will be described in more detail.

As used herein, the term "halogen" means fluorine, chlorine, bromine or iodine, unless stated otherwise.

As used herein, the term 'alkyl' means a straight, cyclic or branched hydrocarbon moiety unless stated otherwise.

The term cycloalkyl, as used herein, unless otherwise indicated, refers to cyclic alkyl including cyclopropyl and the like.

As used herein, the term 'aryl' refers to an aromatic group including phenyl, naphthyl, and the like, unless stated otherwise.

The term 'heterocycloalkyl' as used herein refers to monocyclic or bicyclic or higher cyclic alkyl containing heteroatoms selected from O, N and S unless otherwise indicated. Examples of mono heterocycloalkyl include, but are not limited to, piperidinyl, morpholinyl, thiamorpholinyl, pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, piperazinyl, and the like no.

As used herein, the term 'heteroaryl' means a monocyclic or bicyclic or higher aromatic group containing a heteroatom selected from O, N and S, unless otherwise indicated. Examples of monocyclic heteroaryls are thiazolyl, oxazolyl, thiophenyl, furanyl, pyrrolyl, imidazolyl, isoxazolyl, pyrazolyl, triazolyl, thiadiazolyl, tetrazolyl, oxadizolyl, pyridine 1, pyridazinyl, pyrimidinyl, pyrazinyl and the like, but are not limited to these. Examples of bicyclic heteroaryls include indolyl, benzothiophenyl, benzofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzthiazolyl, benzthiadiazolyl, benzizozolyl, quinolinyl, iso Quinolinyl, purinyl, puropyridinyl, and the like, but are not limited to these.

The compounds according to the invention can also form pharmaceutically acceptable salts. Such pharmaceutically acceptable salts are not particularly limited as long as they form acids that form non-toxic acid addition salts containing pharmaceutically acceptable anions. For example, inorganic acids, such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, hydroiodic acid, etc .; Organic carbon acids such as tartaric acid, formic acid, citric acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, and the like; And acid addition salts formed by sulfonic acids such as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid and the like. Preferred examples of these include acid addition salts formed by sulfuric acid, methanesulfonic acid or hydrochloric acid.

On the other hand, the compounds according to the invention may have an asymmetric carbon center and therefore may exist as R or S isomers, racemic compounds, diastereomeric mixtures, or individual diastereomers, all of these isomers and mixtures being within the scope of the invention. Included.

In addition, solvate and hydrate forms of the compound of formula 1 are also included within the scope of the present invention.

In the imidazopyrazinone derivative compound of Formula 1 of the present invention, preferably, X is oxygen or hydrogen; R ₂ and R ₃ are each independently hydrogen, C _3-14 aryl, C _2-13 heteroaryl, or C _1-3 alkyl substituted with one or more C _3-14 aryl or C _2-13 heteroaryl; R ₁ , R ₄ , R ₅ and R ₆ are each independently hydrogen, C _1-8 alkyl, C _3-8 cycloalkyl, or C _2-7 heterocycloalkyl; R ₇ and R ₈ are each independently hydrogen, —COOR ₉ , C _1-8 alkyl, C _3-8 cycloalkyl, or C _2-7 heterocycloalkyl; The alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are each independently hydrogen, R ₉ , -OH, -OR ₉ , -COR ₉ , -COOH, -COOR ₉ , -CONR ₉ R ₁₀ , -NR ₉ , -NR ₉ CONR ₁₀ R ₁₁ , -CONR ₉ R ₁₀ , -NR ₉ CONR ₁₀ R ₁₁ , -SO ₂ R ₉ , -SO ₂ H and -SOR ₉ ; R ₉ to R ₁₁ are each independently hydrogen, halogen, amino, nitro, carboxy, cyano, sulfyl, hydroxy, C _1-4 alkyl, C _1-4 alkoxy, C _3-6 cycloalkyl, C _{2 -5} heterocycloalkyl, C _3-14 aryl, or C _2-13 heteroaryl.

As each substituent, it is preferable that X is oxygen first.

In addition, R ₂ is C _1-3 alkyl substituted with one or more C _3-14 aryl or C _2-13 heteroaryl, wherein the aryl and heteroaryl are each independently hydrogen, halogen, C _1-4 alkyl, It is preferably substituted with one or more selected from the group consisting of C _1-4 alkoxy, -COO-C _1-4 alkyl. More preferably, R ₃ is C _1-3 alkyl substituted with C _3-14 aryl or C _2-13 heteroaryl, wherein the aryl and heteroaryl are each independently hydrogen, hydroxy, C _1-4 alkyl and It is preferably substituted with one or more selected from the group consisting of C _1-4 alkoxy and C _3-14 aryl.

In addition, R ₃ is methyl substituted with phenyl, naphthyl or indolyl, wherein phenyl, naphthyl and indolyl are each independently one or more selected from the group consisting of hydrogen, hydroxy, methyl, methoxy and phenyl It is preferred to be substituted.

And R ₁ , R ₄ , R ₅ and R ₆ are each independently hydrogen, C _1-8 alkyl, or C _3-8 cycloalkyl, and R ₇ and R ₈ are each independently hydrogen,- Preference is given to COO-C _1-4 alkyl, or C _1-8 alkyl.

In addition, the salt is preferably a hydrochloride salt.

Specific examples of preferred imidazopyrazinone derivative compounds of the present invention are as follows. Also possible are salts, isomers, hydrates or solvates of the following compounds:

1) (S) -N-((S) -2-((5S, 8aR) -5-benzyl-3,6-dioxo-7-phenethylhexahydroimidazo [1,2-a] pyrazine- 1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

2) (S) -N-((S) -1-cyclohexyl-2-((5S, 8aR) -5- (4-hydroxybenzyl) -3,6-dioxo-7-phenethylhexahydro Imidazo [1,2-a] pyrazin-1 (5H) -yl) -2-oxoethyl) -2- (methylamino) propanamide;

3) (S) -N-((S) -2-((5S, 8aR) -5-benzyl-7- (2,2-diphenylethyl) -3,6-dioxohexahydroimidazo [1] , 2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

4) (S) -N-((S) -2-((5R, 8aS) -5-benzyl-3,6-dioxo-7-phenethylhexahydroimidazo [1,2-a] pyrazine- 1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

5) (S) -2-amino-N-((S) -2-((5S, 8aR) -5-benzyl-3,6-dioxo-7-phenethylhexahydroimidazo [1,2- a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) propanamide;

6) (S) -N-((S) -1-cyclohexyl-2-((5S, 8aR) -5- (4-methoxybenzyl) -3,6-dioxo-7-phenethylhexahydro Imidazo [1,2-a] pyrazin-1 (5H) -yl) -2-oxoethyl) -2- (methylamino) propanamide;

7) (S) -N-((S) -1-cyclohexyl-2-((5S, 8aR) -5- (naphthalen-1-ylmethyl) -3,6-dioxo-7-phenethylhexa Hydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -2-oxoethyl) -2- (methylamino) propanamide;

8) (S) -N-((S) -1-cyclohexyl-2-((5S, 8aR) -5- (naphthalen-2-ylmethyl) -3,6-dioxo-7-phenethylhexa Hydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -2-oxoethyl) -2- (methylamino) propanamide;

9) (S) -N-((S) -2-((5S, 8aR) -5- (biphenyl-4-ylmethyl) -3,6-dioxo-7-phenethylhexahydroimidazo [ 1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

10) ((S) -N-((S) -1-cyclohexyl-2-((5S, 8aR) -5- (4-methylbenzyl) -3,6-dioxo-7-phenethylhexahydro Imidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

11) (S) -N-((S) -2-((5S, 8aR) -5-benzyl-3-oxo-7-phenethylhexahydroimidazo [1,2-a] pyrazine-1 (5H ) -Yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

12) (S) -N-((S) -2-((5S, 8aR) -5-benzyl-3,6-dioxo-7- (3-phenylpropyl) hexahydroimidazo [1,2- a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

13) (S) -N-((S) -2-((5S, 8aR) -5-benzyl-7- (4-fluorophenethyl) -3,6-dioxohexahydroimidazo [1, 2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

14) (S) -N-((S) -3,3-dimethyl-1-((5S, 8aR) -5- (naphthalen-1-ylmethyl) -3,6-dioxo-7-phenethyl Hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-oxobutan-2-yl) -2- (methylamino) propanamide;

15) (S) -N-((S) -1-cyclohexyl-2-((2S, 5S, 8aR) -2-methyl-5- (naphthalen-1-ylmethyl) -3,6-dioxo -7-phenethylhexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -2-oxoethyl) -2- (methylamino) propanamide;

16) (S) -N-((S) -2-((5S, 8aR) -7-benzyl-5- (naphthalen-2-ylmethyl) -3,6-dioxohexahydroimidazo [1, 2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

17) (S) -N-((S) -2-((5S, 8aR) -7- (4-chlorobenzyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxohexahydro Imidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

18) (S) -N-((S) -2-((5S, 8aR) -7- (4-methoxybenzyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxohexahexa Hydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

19) (S) -N-((S) -2-((5S, 8aR) -7- (4-methoxyphenethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxo Hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

20) (S) -N-((S) -2-((5S, 8aR) -7- (4-fluorophenethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxo Hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

21) (S) -N-((S) -2-((5S, 8aR) -7- (4-fluorophenethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxo Hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;

22) (S) -N-((S) -2-((5S, 8aR) -7- (4-fluorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahydro Imidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

23) (S) -N-((S) -2-((5S, 8aR) -7- (4-fluorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahydro Imidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;

24) (S) -N-((S) -2-((5S, 8aR) -7- (3-chlorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahydroimide Dazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

25) (S) -N-((S) -2-((5S, 8aR) -7- (2-chlorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahydroimide Dazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

26) (S) -N-((S) -2-((5S, 8aR) -5,7-bis (4-methylbenzyl) -3,6-dioxohexahydroimidazo [1,2-a ] Pyrazin-1 (5H) -yl-3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;

27) (S) -N-((S) -1-((5S, 8aR) -7- (3-chlorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahydroimide Dazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;

28) (S) -N-((S) -2-((5S, 8aR) -7- (4-pyridinemethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxohexahydro Imidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

29) (S) -N-((S) -2-((5S, 8aR) -7- (4-pyridinemethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxohexahydro Imidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;

30) (S) -N-((S) -2-((5S, 8aR) -7- (methyl- (4-methylbenzoate))-5- (naphthalen-2-ylmethyl) -3,6 -Dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

31) (S) -N-((S) -2-((5S, 8aR) -7- (methyl- (4-methylbenzoate))-5- (naphthalen-2-ylmethyl) -3,6 -Dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;

32) (S) -N-((S) -1-((5S, 8aR) -7- (2-chlorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahydroimide Dazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;

33) (S) -N-((S) -3,3-dimethyl-1-((5S, 8aR) -5- (naphthalen-2-ylmethyl) -3,6-dioxo-7-phenethyl Hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-oxobutan-2-yl) -2- (methylamino) propanamide;

34) (S) -N-((S) -2-((5S, 8aR) -7- (4-chlorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahydroimide Dazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

35) (S) -N-((S) -1-((5S, 8aR) -7- (4-chlorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahydroimide Dazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;

36) Methyl 4-(((5S) -5- (biphenyl-4-ylmethyl) -1-((S) -2-cyclohexyl-2-((S) -2- (methylamino) propaneami Acetyl) -3,6-dioxohexahydroimidazo [1,2-a] pyrazin-7 (1H) -yl) methyl) benzoate;

37) (S) -N-((S) -2-((5S, 8aR) -7- (3-chlorophenethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxohexa Hydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;

38) (S) -N-((S) -2-((5S, 8aR) -7- (2-chlorophenethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxohexa Hydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;

39) (S) -N-((S) -2-((5S, 8aR) -7- (2-phenylpropyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxohexahydro Imidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;

40) (2S) -N-((2S) -1-((5S) -5-((1H-indol-3-yl) methyl) -3,6-dioxo-7-pentylhexahydroimidazo [ 1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl-2- (methylamino) propanamide;

41) (2S) -N-((1S) -2-((5S) -5-((1H-indol-3-yl) methyl) -3,6-dioxo-7-pentylhexahydroimidazo [ 1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;

42) ((S) -N-((S) -2-((5S, 8aR) -7- (2-fluorophenethyl) -5- (naphthalen-2-ylmethyl) -3,6-di Oxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;

43) (S) -N-((S) -2-((5S, 8aR) -7- (2-bromophenethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxohexa Hydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;

44) (S) -N-((S) -2-((5S, 8aR) -7- (2-methoxyphenethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxo Hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;

45) (S) -N-((S) -2-((5S, 8aR) -7- (2,3-dihydro-1H-inden-2-yl) -5- (naphthalen-2-ylmethyl ) -3,6-dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino Propanamide;

46) (S) -N-((S) -2-((5S, 8aR) -7- (4-methyl-1,2,3,4-tetrahydronaphthalen-2-yl) -5- (naphthalene -2-ylmethyl) -3,6-dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl)- 2- (methylamino) propanamide; And

47) (S) -N-((S) -2-((5S, 8aR) -7- (1,2,3,4-tetrahydronaphthalen-2-yl) -5- (naphthalen-2-yl Methyl) -3,6-dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methyl Amino) propanamide.

Synthesis of peptide analogs of the libraries of the present invention can be carried out using general schemes for preparing smack / diablo like structure libraries as shown in Schemes 1 and 2. Synthesis of selected peptide analogues of the mono-cyclic template library of the present invention can be accomplished using a solution phase reaction, FlexChem reactor block with 96 well plates or by a syringe reaction. This can be done through. The resin used in the above schemes and examples is conventionally available 3-bromo-1-methoxy-propyl-1-oxyresin (hereinafter referred to as 'bromoacetal resin', CreoSalus) by combinatorial chemical techniques. Mass spectrometry can be performed using Waters' MicroMass ZQ ^™ .

According to the present invention, the intermediates or derivatives of the structure of Formula 1 and the novel smack / diablo-like derivatives can be efficiently constructed by using a solution phase chemical reaction, a solid phase single synthesis method, and an equilibrium synthesis method. Pharmaceutical compositions comprising derivatives, salts, isomers, hydrates or solvates thereof synthesized as an active ingredient, can be used for the prevention of cancer, inflammation, autoimmune diseases and neurodegenerative disorders caused by overexpression of apoptosis inhibiting proteins and It can be used as a therapeutic agent.

The pharmaceutical compositions of the present invention may be formulated according to conventional methods and may be formulated in various oral dosage forms such as tablets, pills, powders, capsules, syrups, emulsions, microemulsions or parenteral such as intramuscular, intravenous or subcutaneous administration. It may be prepared in a dosage form.

When the pharmaceutical composition of the present invention is prepared in the form of an oral dosage form, examples of the carrier used include cellulose, calcium silicate, corn starch, lactose, sucrose, dextrose, calcium phosphate, stearic acid, magnesium stearate and stearic acid. Calcium, gelatin, talc, surfactants, suspending agents, emulsifiers, diluents and the like. When the pharmaceutical composition of the present invention is prepared in the form of an injection, the carrier includes water, saline solution, aqueous glucose solution, pseudo-aqueous solution, alcohol, glycol, ether (e.g. polyethylene glycol 400), oil, fatty acid, fatty acid ester, glycerol Ride, surfactant, a suspending agent, an emulsifier, etc. are mentioned.

Hereinafter, the manufacturing method of the compound which concerns on this invention is demonstrated.

In the following preparation methods and examples, the following abbreviations are used:

Boc-: tert-butoxycarbonyl Cbz-: 2-benzyloxycarbonylamino

DIC: 1,3-diisopropylcarbodiimide DIPEA: N, N-diisopropylethylamine

DMAP: N, N-dimethylaminopyridine DMF: N, N-dimethyl formamide

DMSO: Dimethyl Sulfoxide EA: Ethyl Acetate

EDCI: 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride

HATU: [2- (1H-9-Azabenzotriazol-1-yl) -1,1,3,3-tetramethyluronium hexafluorophosphate] Fmoc-: 9-fluorenyloxycarbonyl

Hex. : Hexane HOBT: N-hydroxybenzotriazole

Mass: Mass Spectrometry Chromatogram MC: Methylene Chloride

MeOH: methineol -OBn: -O-benzyl

THF: Tetrahydrofuran TLC: Thin Layer Chromatography

Phe: Phenylalanine Phe (4-OH): 4-hydroxyphenylalanine

Tle: t-butylglycine Chg: cyclohexylglycine

Bip: Biphenylalanine Ala: Alanine

Nal: naphthylalanine Phe: phenylalanine

MeAla: Methylalanine Trp: Tryptophan

Tyr: Tyrosine Ser: Serine

Gly: glycine

The compound of formula 1 according to the present invention can be prepared according to the solid phase synthesis method shown in Scheme 1 below.

Scheme 1

Wherein R ₂ to R ₇ are the same as defined in Chemical Formula 1.

)의 표시는 고분자 중합체 형태의 고체 지지체(solid support)를 나타내며, 이 고체 지지체의 핵심 작용기를 나타내었다. PG는 보호그룹 (protection group)을 표시한다. In Scheme 1 above (

Denotes a solid support in the form of a high molecular polymer and represents the key functional groups of the solid support. PG denotes a protection group.

The present invention can be prepared through a general method of synthesizing Chemical Formula 1 through the solid phase synthesis process of Scheme 1 illustrated above. Depending on the residue of the R ₃ group, the five-step cyclization reaction may proceed simultaneously with the thresin reaction and the cyclization reaction in step 6. A more generalized reaction process is illustrated in the following step reaction method.

First step

Bromoacetal resin (1.0 equiv) is placed in a 96 well Robinson block (FlexChem) or a syringe with a filtration filter and the solution in which amine (8.0-12.0 equiv) is dissolved in DMSO is added to the syringe or Suck Shake these 96 well Robinson blocks or syringes in a 50-70 ° C rotary oven for 10-20 hours. After cooling the Robinson block or syringe to room temperature, the suspension is filtered and the resin is washed several times with DMF, MeOH, and MC, respectively.

2nd step

After swelling the resin obtained in the above step with DMF, the DMF solution was filtered, and Fmoc-amino acid (2.0-4.0 equivalents), HATU (2.0-4.0 equivalents) and DIPEA (4.0-8.0 equivalents) were dissolved in DMF in the resin. The solution is added dropwise and shaken at room temperature for 4-8 hours. The suspension is filtered and the resin washed several times with DMF, MeOH and MC, respectively.

After swelling the resin obtained in the above process in DMF, the DMF solution is filtered, piperidine / DMF is added dropwise to the resin and shaken for 1 to 2 hours at room temperature. The suspension is filtered and the resin washed several times with DMF, MeOH, and MC, respectively.

3rd step

After swelling the resin obtained in the above step with DMF, the DMF solution was filtered, and the resin was subjected to Fmoc-amino acid (2.0 to 4.0 equivalents), DIC (2.0 to 4.0 equivalents), HOBT (2.0 to 4.0 equivalents) and DIPEA (4.0 to 8.0 equivalents). Equivalent solution) is added dropwise to the solution of DMF and shaken at room temperature for 4.0-8.0 hours. The suspension is filtered and the resin washed several times with DMF, MeOH and MC, respectively.

After swelling the resin obtained in the above process in DMF, the DMF solution is filtered, piperidine / DMF is added dropwise to the resin and shaken for 1 to 2 hours at room temperature. The suspension is filtered and the resin washed several times with DMF, MeOH, and MC, respectively.

4th step

After swelling the resin obtained in the above step with DMF, the DMF solution was filtered, and the resin was subjected to Fmoc-amino acid (2.0 to 4.0 equivalents), DIC (2.0 to 4.0 equivalents), HOBT (2.0 to 4.0 equivalents) and DIPEA (4.0 to 8.0 equivalents). Equivalent) dissolved in DMF dropwise and shake for 4-8 hours at room temperature. The suspension is filtered and the resin washed several times with DMF, MeOH, and MC, respectively.

In addition, the resin obtained in the above process was swollen in DMF, and then the DMF solution was filtered, piperidine / DMF was added dropwise to the resin, and the mixture was shaken at room temperature to filter the suspension, and the resin was several times washed with DMF, MeOH, and MC. Can be washed

5th step

The resin obtained in the above step is placed in a seal tube and formic acid is added dropwise. Heat at 70 ~ 80 ^o C for 3 ~ 5 hours, and reduce to room temperature. Celite filtration removes the solid support remaining after the reaction and distillates under reduced pressure to obtain a residue. The residue obtained through this process is purified using column chromatography.

The compound obtained in the above process is dissolved in piperidine / MC and stirred at room temperature. After the reaction, the solution is concentrated under reduced pressure and purified by column chromatography.

6th step

Boc-protected amino acids (1.0-2.0 equiv), EDCI (1.0-2.0 equiv), HOBT (1.0-2.0 equiv), and DIPEA (4.0-8.0 equiv) are dissolved in MC and stirred at room temperature for 10-20 minutes. . To this solution is added the amine (1.0 equiv) obtained in step (5) and stirred at room temperature for 3-5 hours. After the reaction, the solution is anhydrous treated with magnesium sulfate, concentrated under reduced pressure and purified by column chromatography.

The compound obtained in the above process is dissolved in HCl / dioxane or formic acid and stirred at room temperature for 1-2 hours. After the reaction, the solution is concentrated under reduced pressure and recrystallized with methanol and EA to obtain the title compound.

In addition, the compound of Formula 1 according to the present invention may be prepared according to the solution phase synthesis method of Scheme 2 below.

Scheme 2

Wherein R ₂ to R ₇ are the same as defined in Chemical Formula 1, and PG represents a protection group.

The present invention can be prepared through a general method of synthesizing a derivative of Formula 1 through the solution phase synthesis process of Scheme 2 illustrated above. A more generalized reaction process is illustrated in the following step reaction method.

First step

The amine (1.0 equiv) and dimethylacetaldehyde (1.0 equiv) are dissolved in methanol and then stirred at room temperature for 1 hour. NaCNBH ₃ (1.8-2.2 equiv) is added to this solution followed by the dropwise addition of acetic acid in catalytic amounts. The reaction solution is stirred at room temperature for 4-16 hours, concentrated under reduced pressure and purified by column chromatography.

2nd step

Fmoc-protected amino acids (1.5-2.0 equiv), HATU (1.5-2.0 equiv) and DIPEA (2.0-4.0 equiv) were dissolved in MC and stirred at room temperature for 10-20 minutes. To this solution is added the amine (1.0 equiv) obtained in step (1) and stirred at room temperature for 3-5 hours. After the reaction, the solution is concentrated under reduced pressure and purified by column chromatography.

The obtained compound is dissolved in piperidine / MC and stirred at room temperature for 1 to 2 hours. After the reaction, the solution is concentrated under reduced pressure and purified by column chromatography.

3rd step

Fmoc-protected amino acids (1.5-2.0 equiv), EDCI (1.5-2.0 equiv), HOBT (1.5-2.0 equiv) and DIPEA (2.0-4.0 equiv) were dissolved in MC and stirred at room temperature for 4-16 hours. . To this solution is added the amine (1.0 equiv) obtained in step (2) and stirred at room temperature for 4-16 hours. After the reaction, the solution is concentrated under reduced pressure and purified by column chromatography.

The obtained compound is dissolved in piperidine / MC and stirred at room temperature for 1 to 2 hours. After the reaction, the solution is concentrated under reduced pressure and purified by column chromatography.

4th step

Fmoc-protected amino acids (1.5-2.0 equiv), EDCI (1.5-2.0 equiv), HOBT (1.5-2.0 equiv) and DIPEA (2.0-4.0 equiv) were dissolved in MC and stirred at room temperature for 4-16 hours. To this solution is added the amine (1.0 equiv) obtained in step (3) and stirred at room temperature for 4-16 hours. After the reaction, the solution is concentrated under reduced pressure and purified by column chromatography.

5th step

Formic acid is added dropwise to the compound obtained in the fourth step. Stir at 70 ~ 100 ^o C for 2 ~ 4 hours, concentrate and purify using column chromatography.

The compound obtained in the above process is dissolved in piperidine / MC and stirred at room temperature for 1 to 3 hours. After the reaction, the solution is concentrated under reduced pressure and purified by column chromatography.

6th step

Boc-protected amino acids (1.5-2.0 equiv), EDCI (1.5-2.0 equiv), HOBT (1.5-2.0 equiv) and DIPEA (2.0-4.0 equiv) were dissolved in MC and stirred at room temperature for 10-20 minutes. To this solution is added the amine (1.0 equiv) obtained in step (5) and stirred at room temperature for 3-5 hours. After the reaction, the solution is concentrated under reduced pressure, which is anhydrous treated with magnesium sulfate, concentrated under reduced pressure and purified by column chromatography.

The compound obtained in the above process is dissolved in HCl / dioxane or formic acid and stirred at room temperature for 1 to 3 hours. After the reaction, the solution is concentrated under reduced pressure and recrystallized with methanol and EA to obtain the title compound.

Example

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited by the following examples.

Example 1: (S) -N-((S) -2-((5S, 8aR) -5-benzyl-3,6-dioxo-7-phenethylhexahydroimidazo [1,2-a] Preparation of pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide hydrochloride

Step (1): solid 2-ethoxy-2-methoxy- N Preparation of Phenethylethanamine

Bromoacetal resin (1.0 g, 1.5 mmol) was placed in a syringe with a filtration filter, soaking a solution in which phenethylamine (1.88 mL, 15.0 mmol) was dissolved in DMSO (8 mL). The syringe was shaken for 16 hours in a 60 ° C rotary oven. After the syringe was cooled to room temperature, the suspension was filtered and the resin washed three times with DMF, MeOH, and MC, respectively. After drying in vacuo, the presence of the secondary amine was verified by a chronolin test to give the title compound.

Step (2): solid phase (9 H -Floren-9-yl) methyl (2 S Preparation of) -1-((2-ethoxy-2-methoxyethyl) (phenethyl) amino) -1-oxo-3-phenylpropane-2-ylcarbamate

After swelling the compound (1.5 mmol) obtained in step (1) with DMF, the DMF solution was filtered, and Fmoc-Phe-OH (1.75 g, 4.5 mmol), HATU (1.75 g, 4.5 mmol) and DIPEA ( 1.57mL, 9.0mmol) was sucked into the solution dissolved in DMF (8mL) and shaken for 6 hours at room temperature. The suspension was filtered and the resin washed three times with DMF, MeOH, and MC each. After drying in vacuo, the presence of the secondary amine was verified by a chronolin test to give the title compound.

Step (3): solid phase (2 S ) -2-amino- N -(2-ethoxy-2-methoxyethyl)- N Of Phenethyl-3-phenylpropanamide

The compound (1.5 mmol) obtained in step (2) was swollen in DMF, and then the DMF solution was filtered, 25% piperidine / DMF (8.0 mL) was sucked into the resin, and then shaken at room temperature for 1 hour. The suspension was filtered and the resin washed three times with DMF, MeOH, and MC each. Drying in vacuo followed by a Kaiser test to verify the presence of the primary amine to afford the title compound.

Step (4): Solid Phase (9 H -Floren-9-yl) methyl 2-((2 S Preparation of) -1-((2-ethoxy-2-methoxyethyl) (phenethyl) amino) -1-oxo-3-phenylpropan-2-ylamino) -2-oxoethylcarbamate

After swelling the compound (1.5 mmol) obtained in step (3) in DMF, the DMF solution was filtered, and Fmoc-Gly-OH (1.33 g, 4.5 mmol), DIC (0.7 mL, 4.5 mmol) HOBT (608 mg) was added to the resin. , 4.5 mmol) and DIPEA (1.57 mL, 9.0 mmol) were sucked into the solution dissolved in DMF (8 mL) and shaken for 6 hours at room temperature. The suspension was filtered and the resin washed three times with DMF, MeOH and MC. Drying in vacuo followed by a Kaiser test to verify the presence of the primary amine to afford the title compound.

Step (5): solid phase (2 S )-(2-aminoacetamido)- N -(2-ethoxy-2-methoxyethyl)- N Of Phenethyl-3-phenylpropanamide

The compound (1.5 mmol) obtained in step (4) was swollen in DMF, and then the DMF solution was filtered, 25% piperidine / DMF (8.0 mL) was sucked into the resin, and then shaken at room temperature for 1 hour. The suspension was filtered and the resin washed three times with DMF, MeOH, and MC each. Drying in vacuo followed by a Kaiser test to verify the presence of the primary amine to afford the title compound.

Step (6): solid phase (9 H -Floren-9-yl) methyl (1 S , 7 S Preparation of 7-benzyl-1-cyclohexyl-11-methoxy-2,5,8-trioxo-9-phenethyl-12-oxa-3,6,9-triazatetradecylcarbamate

After swelling the compound (1.5 mmol) obtained in step (5) in DMF, the DMF solution was filtered, and Fmoc-Chg-OH (1.70 g, 4.5 mmol), DIC (0.7 mL, 4.5 mmol), HOBT ( 608mg, 4.5mmol) and DIPEA (1.57mL, 9.0mmol) were sucked in the solution dissolved in DMF (8mL) and shaken for 6 hours at room temperature. The suspension was filtered and the resin washed three times with DMF, MeOH, and MC each. Drying in vacuo followed by a Kaiser test to verify the presence of the primary amine to afford the title compound.

Step (7): (9H-Floren-9-yl) methyl (S) -2-((5S, 8aR) -5-benzyl-3,6-dioxo-7-phenethylhexahydroimidazo [1 Preparation of, 2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethylcarbamate

The compound (1.5 mmol) obtained in step (6) was added to a sealed tube, and 20 mL of formic acid was added dropwise thereto, followed by stirring at 80 ° C. for 4 hours. After the reaction, the reaction solution was cooled to room temperature, filtered under reduced pressure using a celite filter, and then filtered through MC / MeOH (v / v = 9/1). The filtered solution was concentrated under reduced pressure and purified by column chromatography to give the title compound as a yellow oil (383 mg, 36%).

¹ H NMR (300 MHz, CDCl ₃ ): 7.75 (d, 2H), 7.59 (d, 2H), 7.39-7.05 (m, 12H), 6.54 (d, 2H), 5.82 (br, 1H), 5.49 (br , 1H), 5.44-5.36 (m, 1H), 4.99 (br, 1H), 4.95-4.86 (m, 1H), 4.76 (br, 1H), 4.44-4.36 (m, 2H), 4.23-4.13 (m , 2H), 3.80-3.76 (m, 1H), 3.34-3.12 (m, 3H), 1.73-1.61 (m, 5H), 1.17-0.95 (m, 5H)

MS (ESI ⁺ , m / z): 711 [M + H] ⁺

Step (8): (5S, 8aR) -1-((S) -2-Amino-2-cyclohexylacetyl) -5-benzyl-7-phenethyltetrahydroimidazo [1,2-a] pyrazine- Preparation of 3,6 (5H, 7H) -dione

The compound (300mg, 0.422mmol) obtained in step (7) was dissolved in a piperidine / MC (v / v = 1/9) solution and stirred at room temperature for 1 hour. After reaction the solution was concentrated under reduced pressure and purified by column chromatography to give the title compound as a white solid (150 mg, 72%).

MS (ESI ⁺ , m / z): 489 [M + H] ⁺

Step (9): t-butyl (S) -1-((S) -2-((5S, 8aR) -5-benzyl-3,6-dioxo-7-phenethylhexahydroimidazo [1, 2-a] Pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethylamino) -1-oxopropan-2-yl (methyl) carbamate

Boc-MeAla-OH (93mg, 0.459mmol) was dissolved in MC (10mL), and then EDC (87mg, 0.459mmol), HOBT (62mg, 0.459mmol) and DIPEA (0.16mL, 0.918mmol) were added dropwise and 15 at room temperature. Stir for minutes. To the reaction solution was added dropwise the compound (150mg, 0.306mmol) obtained in step (8) and stirred at room temperature for 4 hours. After the reaction, sodium bicarbonate (20 mL) was added dropwise to the solution to separate the organic layer, and the organic layer was dried over sodium sulfate, filtered under reduced pressure and distilled under reduced pressure. The obtained residue was purified by column chromatography to give the title compound as a white solid (185 mg, 90%).

¹ H NMR (300MHz, CDCl ₃ ): 7.24-6.75 (m, 10H), 5.75 (d, 1H), 5.42 (d, 1H), 4.83 (m, 1H), 4.57 (m, 1H), 4.28 (m , 2H), 4.13-4.03 (m, 2H), 3.72 (s, 3H), 3.60-3.08 (m, 6H), 2.78 (s, 3H), 1.70-0.90 (m, 19H)

MS (ESI ⁺ , m / z): 674 [M + H] ⁺

Step (10): (S) -N-((S) -2-((5S, 8aR) -5-benzyl-3,6-dioxo-7-phenethylhexahydroimidazo [1,2-a ] Preparation of pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide hydrochloride

The compound (180 mg, 0.267 mmol) obtained in step (9) was dissolved in 4M HCl / dioxane solution (10 mL) and stirred at room temperature for 1.5 hours. After the reaction, the solution was concentrated under reduced pressure and recrystallized using MC and ethyl acetate to give the title compound as a white solid (85 mg, 52%).

¹ H NMR (300MHz, CD ₃ OD): 8.34 (br, 1H), 7.28-6.88 (m, 10H), 5.74 (s, 1H), 5.39 (s, 1H), 5.23 (d, 1H), 4.80 ( br, 1H), 4.34-4.33 (m, 1H), 4.19-4.07 (m, 2H), 3.90-3.87 (m, 2H), 3.67-3.65 (m, 2H), 3.24-3.06 (m, 2H), 2.75-2.61 (m, 6H), 1.76-1.12 (m, 13H)

MS (ESI ⁺ , m / z): 574 [M + H] ⁺

S) -N-((S) -2-((5S, 8aR) -5-benzyl-3,6-dioxo-7-phenethylhexahydroimidazo [1,2-a] synthesized in Example 1 ] Pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide hydrochloride may be synthesized through the solution phase preparation method of Example 1-1 below.

Example 1-1: (S) -N-((S) -2-((5S, 8aR) -5-benzyl-3,6-dioxo-7-phenethylhexahydroimidazo [1,2- a] Liquid phase preparation of pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide hydrochloride

Step (1): Preparation of 2,2-dimethoxy-N-phenethylethanamine

2,2-dimethoxyacetaldehyde (5.0 g, 48.0 mmol) and phenethylamine (6 mL, 48.0 mmol) were dissolved in methanol (250 mL). After stirring at room temperature for 1 hour, sodium cyanoborohydride (6.0 g, 96.0 mmol) and acetic acid (1 mL) were added thereto, and the mixture was stirred at room temperature for 16 hours. The residue obtained after concentration of the reaction was purified by column chromatography to give the title compound as a clear oil (6.1 g, 61%).

¹ H NMR (300 MHz, CDCl ₃ ): 7.30-7.17 (m, 5H), 4.69 (t, 1H, J = 5.37), 3.39 (s, 6H), 2.75-2.63 (m, 6H), 1.84 (s, 1H)

MS (ESI ⁺ , m / z): 209 [M] ⁺

Step (2): (S)-(9H-Floren-9-yl) methyl 1-((2,2-dimethoxyethyl) (phenethyl) amino) -1-oxo-3-phenylpropane-2- Preparation of Ilcarbamate

Fmoc-Phe-OH (7.22 g, 18.64 mmol), HATU (7.08 g, 18.64 mmol), and DIPEA (6.49 mL, 37.29 mmol) were dissolved in MC (50 mL) and stirred at room temperature for 15 minutes. The compound (3.0 g, 14.34 mmol) obtained in the step (1) was added dropwise to the reaction solution, followed by stirring at room temperature for 4 hours. After the reaction, the organic solution was washed with saturated sodium hydrogen carbonate solution, dried over sodium sulfate and filtered. The filtered organic solvent layer was concentrated under reduced pressure and the residue obtained was purified by column chromatography to give the title compound as a clear oil (6.14 g, 71%).

MS (ESI ⁺ , m / z): 579 [M + H] ⁺

Step (3): Preparation of (S) -2-amino-N- (2,2-dimethoxyethyl) -N-phenethyl-3-phenylpropanamide

The compound (1.0 g, 1.728 mmol) obtained in step (2) was dissolved in dichloromethane (20 mL), and piperidine (2 mL) was added thereto, followed by stirring for one hour. The reaction solution was concentrated and purified by column chromatography to give the title compound as a yellow oil (458 mg, 74%).

MS (ESI ⁺ , m / z): 357 [M + H] ⁺

Step (4): (S)-(9H-Floren-9-yl) methyl 2- (1-((2,2-dimethoxyethyl) (phenethyl) amino) -1-oxo-3-phenylpropane Preparation of 2-ylamino) -2-oxoethylcarbamate

Fmoc-Gly-OH (1.08 g, 3.64 mmol), EDC (697 mg, 3.64 mmol), HOBT (491 mg, 3.64 mmol), and DIPEA (1.26 mL, 7.28 mmol) were dissolved in MC (20 mL), followed by room temperature for 15 minutes. Stirred at. The compound (1.0 g, 2.8 mmol) obtained in the step (3) was added dropwise to the reaction solution, followed by stirring at room temperature for 4 hours. After the reaction, the organic solution was washed with saturated sodium hydrogen carbonate solution, dried over sodium sulfate and filtered. The filtered organic solvent layer was concentrated under reduced pressure and the residue obtained was purified by column chromatography to give the title compound as a clear oil (1.35 g, 76%).

MS (ESI ⁺ , m / z): 636 [M + H] ⁺

Step (5): Preparation of (S) -2- (2-aminoacetamido) -N- (2,2-dimethoxyethyl) -N-phenethyl-3-phenylpropanamide

The compound (4.0 g, 6.29 mmol) obtained in step (4) was dissolved in dichloromethane (100 mL), and piperidine (10 mL) was added thereto, followed by stirring for one hour. The reaction solution was concentrated and purified by column chromatography to give the title compound as a yellow oil (2.4 g, 80%).

MS (ESI ⁺ , m / z): 478 [M + H] ⁺

Step (6): (9H-Floren-9-yl) (7S, 13S) -7-benzyl-13-cyclohexyl-3-methoxy-6,9,12-trioxo-5-phenethyl-2 Preparation of oxa-5,8,11-triaztridecane-13-ylcarbamate

The compound (1.2 g, 2.902 mmol) obtained in step (5) was dissolved in dichloromethane (10 mL), and then Fmoc-Chg-OH (1.211 g, 3.192 mmol), EDCI (612 mg, 3.192 mmol), HOBT (431 mg, 3.192 mmol), and DIPEA (1.1 mL, 6.384 mmol) were dissolved in dichloromethane (50 mL) and stirred at room temperature for 3 hours. After the reaction, 5% citric acid solution and saturated sodium bicarbonate were added dropwise to the solution, followed by extraction to separate the organic layer. The organic layer was dried over sodium sulfate, filtered under reduced pressure, and distilled under reduced pressure. The obtained residue was purified by column chromatography to give the title compound as a white solid (1.351 g, 60%).

MS (ESI ⁺ , m / z): 775 [M + H] ⁺

Step (7): (9H-Floren-9-yl) methyl (1S) -2-((5S) -5-benzyl-3,6-dioxo-7-phenethylhexahydroimidazo [1,2 -a] Preparation of pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethylcarbamate

The compound (100 mg, 0.13 mmol) obtained in step (6) was dissolved in formic acid (5 mL) and stirred at 100 ° C. for 3 hours in a seal tube. The reaction was concentrated and purified by column chromatography to give the title compound as a yellow solid (29 mg, 32%).

¹ H NMR (300 MHz, CDCl ₃ ): 7.75 (d, J = 7.4 Hz, 2H), 7.59 (d, J = 7.1 Hz, 2H), 7.34-7.11 (m, 12H), 6.89-6.88 (m, 2H ), 5.75 (br, 1H), 5.40-5.38 (m, 2H), 4.92 (br, 1H), 4.59 (br, 1H), 4.42-4.03 (m, 2H), 4.23-4.03 (m, 4H), 3.63-3.08 (m, 6H), 2.64-2.60 (m, 2H), 1.25-1.01 (m, 12H)

MS (ESI ⁺ , m / z): 711 [M + H] ⁺

Step (8): (5S) -1-((S) -2-Amino-2-cyclohexylacetyl) -5-benzyl-7-phenethyltetrahydroimidazo [1,2-a] pyrazine-3, Preparation of 6 (5H, 7H) -dione

The compound (300mg, 0.42mmol) obtained in step (7) was dissolved in dichloromethane (20mL), and piperidine (2mL) was added thereto, followed by stirring for one hour. The reaction was concentrated and purified by column chromatography to give the title compound as a white solid (150 mg, 72%).

MS (ESI ⁺ , m / z): 489 [M + H] ⁺

Step (9): t-butyl (2S) -1-((1S) -2-((5S) -5-benzyl-3,6-dioxo-7-phenethylhexahydroimidazo [1,2- a] Preparation of pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethylamino) -1-oxopropan-2-yl (methyl) carbamate

Boc-MeAla-OH (93mg, 0.459mmol) was dissolved in MC (10mL), and then EDCI (87mg, 0.459mmol), HOBT (62mg, 0.459mmol), and DIPEA (0.16mL, 0.918mmol) were added dropwise and at room temperature. Stir for 15 minutes. To the reaction solution was added dropwise the compound (150mg, 0.306mmol) obtained in step (8) and stirred at room temperature for 4 hours. After the reaction, 5% citric acid solution and saturated sodium bicarbonate were added dropwise to the solution, followed by extraction to separate the organic layer. The organic layer was dried over sodium sulfate, filtered under reduced pressure and distilled under reduced pressure. The obtained residue was purified by column chromatography to give the title compound as a white solid (185 mg, 90%).

¹ H NMR (300MHz, CDCl ₃ ): 7.24-6.75 (m, 10H), 5.75 (d, J = 4.3 Hz, 1H), 5.42 (m, 1H), 4.83 (m, 1H), 4.57 (m, 1H ), 4.28 (m, 2H), 4.13-4.03 (m, 2H), 3.72 (s, 3H), 3.60-3.08 (m, 6H), 2.78 (s, 3H), 1.70-0.91 (m, 19H)

MS (ESI ⁺ , m / z): 674 [M + H] ⁺

Step (10): (S) -N-((S) -2-((5S, 8aR) -5-benzyl-3,6-dioxo-7-phenethylhexahydroimidazo [1,2-a ] Preparation of pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide hydrochloride

The compound (180 mg, 0.267 mmol) obtained in step (9) was dissolved in 4M HCl / dioxane solution (10 mL) and stirred at room temperature for 1.5 hours. After the reaction, the solution was concentrated under reduced pressure and recrystallized using MC and ethyl acetate to give the title compound as a white solid (85 mg, 52%).

¹ H NMR (300MHz, MeOD-d6): 8.34 (br, 1H), 7.28-6.88 (m, 10H), 5.74 (s, 1H), 5.39 (s, 1H), 5.23 (d, J = 4.6 Hz, 1H), 4.80 (br, 1H), 4.34-4.33 (m, 1H), 4.19-4.07 (m, 2H), 3.90-3.87 (m, 2H), 3.67-3.65 (m, 2H), 3.24-3.06 ( m, 2H), 2.75-2.61 (m, 6H), 1.76-1.12 (m, 13H)

MS (ESI ⁺ , m / z): 574 [M + H] ⁺

Example 2: (S) -N-((S) -1-cyclohexyl-2-((5S, 8aR) -5- (4-hydroxybenzyl) -3,6-dioxo-7-phenethyl Preparation of Hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -2-oxoethyl) -2- (methylamino) propanamide hydrochloride

The title compound was obtained in the same manner as in Example 1 except that Fmoc-Tyr (OtBu) -OH was used instead of Fmoc-Phe-OH of Example 2 (198 mg, 74%).

Example 3: (S) -N-((S) -2-((5S, 8aR) -5-benzyl-7- (2,2-diphenylethyl) -3,6-dioxohexahydroimidazo Preparation of [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide hydrochloride

The title compound was obtained as a white solid (205 mg, 80%) in the same manner as in Example 1 except that 2,2-diphenylethylamine was used instead of the phenethylamine of Step 1 in Example 1.

¹ H NMR (300MHz, CDCl ₃ ): 8.62 (d, 1H), 8.29-8.25 (m, 1H), 7.28-7.03 (m, 15H), 5.57 (br, 1H), 5.36 (br, 1H), 5.00 (d, 1H), 4.71 (d, 1H), 4.32 (t, 1H), 4.16-4.06 (m, 1H), 4.01-3.75 (m, 5H), 3.52-3.46 (m, 1H), 3.25-3.17 (m, 1H), 3.07 (d, 1H), 2.99-2.86 (m, 1H), 1.67 (m, 6H), 1.35 (d, 3H), 0.96-0.88 (m, 4H), 2.46 (s, 3H )

MS (ESI ⁺ , m / z): 650 [M + H] ⁺

Example 4: (S) -N-((S) -2-((5R, 8aS) -5-benzyl-3,6-dioxo-7-phenethylhexahydroimidazo [1,2-a] Preparation of pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide hydrochloride

The title compound was obtained as a white solid through the same method as Example 1 except for using (D) -Fmoc-Phe-OH instead of Fmoc-Phe-OH in Example 1 (16.5 mg, 47%). .

¹ H NMR (300 MHz, CD ₃ OD): 9.18-8.79 (m, 2H), 8.60 (br, 1H), 8.32 (br, 1H), 7.35-7.13 (m, 8H), 6.90 (br, 2H), 5.70 (br, 1H), 5.49 (br, 1H), 5.03 (br, 1H), 4.77 (br, 1H), 4.31 (m, 1H), 4.16 (br, 2H), 3.85 (m, 1H), 3.62 -3.60 (m, 1H), 3.41 (m, 1H), 3.31 (s, 3H), 3.25-3.21 (m, 2H), 3.05-2.96 (m, 1H), 1.66 (m, 6H), 1.33 (br , 3H), 1.10-0.90 (m, 5H)

MS (ESI ⁺ , m / z): 588 [M + H] ⁺

Example 5: (S) -2-amino-N-((S) -2-((5S, 8aR) -5-benzyl-3,6-dioxo-7-phenethylhexahydroimidazo [1, 2-a] Pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) propanamide hydrochloride

The title compound was obtained as a white solid through the same method as Example 1 except for using Boc-Ala-OH instead of Boc-MeAla-OH in Example 1 (48 mg, 83%).

¹ H NMR (300MHz, CD ₃ OD): 8.45 (d, 1H), 8.29-8.17 (m, 3H), 7.36-7.14 (m, 8H), 6.92-6.88 (m, 2H), 5.70 (br, 1H ), 5.51 (br, 1H), 5.03 (d, 1H), 4.77 (d, 1H), 4.28 (t, 1H), 4.06-3.92 (m, 3H), 3.42-3.39 (m, 1H), 3.28- 3.22 (m, 2H), 3.032.94 (m, 1H), 1.66 (br, 6H), 1.19 (d, 3H), 1.10-1.01 (m, 5H)

MS (ESI ⁺ , m / z): 560 [M + H] ⁺

Example 6: (S) -N-((S) -1-cyclohexyl-2-((5S, 8aR) -5- (4-methoxybenzyl) -3,6-dioxo-7-phenethyl Preparation of Hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -2-oxoethyl) -2- (methylamino) propanamide hydrochloride

The title compound was obtained as a white solid through the same method as Example 1 except for using Fmoc-Phe (4-OMe) -OH instead of Fmoc-Phe-OH in Example 1 (36 mg, 74%). .

¹ H NMR (300MHz, CD ₃ OD): 8.29 (br, 1H), 7.20-7.12 (m, 5H), 6.87-6.75 (m, 4H), 5.68 (br, 1H), 5.51 (s, 1H), 5.31 (br, 1H), 5.24-5.22 (m, 1H), 4.33 (br, 1H), 4.18-4.13 (m, 2H), 3.88-3.57 (m, 6H), 3.09-2.82 (m, 2H), 2.58 (s, 3H), 1.77-1.60 (m, 6H), 1.52 (s, 3H), 1.31-0.97 (m, 5H)

MS (ESI ⁺ , m / z): 604 [M + H] ⁺

Example 7: (S) -N-((S) -1-cyclohexyl-2-((5S, 8aR) -5- (naphthalen-1-ylmethyl) -3,6-dioxo-7-pen Preparation of ethyl hexahydroimidazo [1,2-a] pyrazine-1 (5H) -yl) -2-oxoethyl) -2- (methylamino) propanamide hydrochloride

The title compound was obtained as a white solid (50 mg, 15%) by the same method as Example 1 except for using Fmoc-1-Nal-OH instead of Fmoc-Phe-OH in Example 1.

¹ H NMR (300MHz, CD ₃ OD): 9.38 (br, 1H), 8.83 (br, 1H), 8.61 (d, 1H), 6.30 (d, 1H), 7.96-7.84 (m, 3H), 7.59- 7.44 (m, 3H), 7.01-6.73 (m, 5H), 5.84 (br, 1H), 5.65 (br, 1H), 5.20 (br, 1H), 4.94 (br, 1H), 4.31 (t, 1H) , 4.11-4.02 (m, 2H), 3.93-3.69 (m, 2H), 3.51-3.17 (m, 7H), 2.45 (s, 3H), 1.66 (br, 6H), 1.36 (d, 3H), 1.06 (br, 4H)

MS (ESI ⁺ , m / z): 624 [M + H] ⁺

Example 8: (S) -N-((S) -1-cyclohexyl-2-((5S, 8aR) -5- (naphthalen-2-ylmethyl) -3,6-dioxo-7-pen Preparation of ethyl hexahydroimidazo [1,2-a] pyrazine-1 (5H) -yl) -2-oxoethyl) -2- (methylamino) propanamide hydrochloride

The title compound was obtained as a white solid by the same method as Example 1 except for using Fmoc-2-Nal-OH instead of Fmoc-Phe-OH in Example 1 (50 mg, 15%).

¹ H NMR (300MHz, CD ₃ OD): 9.30 (br, 1H), 8.82 (br, 1H), 8.62 (d, 1H), 8.35 (br, 1H), 8.12-7.87 (m, 3H), 7.97- 7.26 (m, 4H), 6.88 (br, 3H), 6.74-6.72 (m, 2H), 6.36 (br, 1H), 6.03 (br, 1H), 5.15 (br, 1H), 4.88 (br, 1H) , 4.40-4.30 (m, 1H), 4.12-3.79 (m, 4H), 3.50-3.39 (m, 2H), 3.26-3.05 (m, 3H), 2.45 (s, 3H), 1.69 (br, 6H) , 1.35 (d, 3H), 1.09 (br, 4H)

MS (ESI ⁺ , m / z): 624 [M + H] ⁺

Example 9: (S) -N-((S) -2-((5S, 8aR) -5- (biphenyl-4-ylmethyl) -3,6-dioxo-7-phenethylhexahydroimide Preparation of Dazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide hydrochloride

The title compound was obtained as a white solid (35 mg, 4%) by the same method as Example 1 except for using Fmoc-1-Nal-OH instead of Fmoc-Phe-OH in Example 1.

MS (ESI ⁺ , m / z): 650 [M + H] ⁺

Example 10: ((S) -N-((S) -1-cyclohexyl-2-((5S, 8aR) -5- (4-methylbenzyl) -3,6-dioxo-7-phenethyl Preparation of Hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide hydrochloride

The title compound was obtained as a white solid through the same method as Example 1 except for using Fmoc-Phe (4-Me) -OH instead of Fmoc-Phe-OH in Example 1 (37 mg, 4%). .

MS (ESI ⁺ , m / z): 588 [M + H] ⁺

Example 11: (S) -N-((S) -2-((5S, 8aR) -5-benzyl-3-oxo-7-phenethylhexahydroimidazo [1,2-a] pyrazine-1 Preparation of (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide hydrochloride

Step (1): Preparation of 2,2-dimethoxy-N-phenethylethanamine

2,2-dimethoxyacetaldehyde (5.0 g, 48.0 mmol) and phenethylamine (6 mL, 48.0 mmol) were dissolved in methanol (250 mL). After stirring at room temperature for 1 hour, sodium cyanoborohydride (6.0 g, 96.0 mmol) and acetic acid (1 mL) were added thereto, and the mixture was stirred at room temperature for 16 hours. The reaction was concentrated and the residue obtained was purified by column chromatography to give the title compound as a clear oil (6.1 g, 61%).

¹ H NMR (300 MHz, CDCl ₃ ): 7.30-7.17 (m, 5H), 4.69 (t, 1H, J = 5.37), 3.39 (s, 6H), 2.75-2.63 (m, 6H), 1.84 (s, 1H)

MS (ESI ⁺ , m / z): 209 [M] ⁺

Step (2): (S)-(9H-Floren-9-yl) methyl 1-((2,2-dimethoxyethyl) (phenethyl) amino) -3-phenylpropan-2-ylcarbamate Produce

The compound (600 mg, 1.62 mmol) obtained in step (1) was dissolved in methanol (10 mL). After stirring at room temperature for 1 hour, sodium cyanoborohydride (203mg, 3.23mmol) and acetic acid (0.1mL) were added thereto, and the mixture was stirred at room temperature for 16 hours. The reaction was concentrated and the residue obtained was purified by column chromatography to give the title compound as a clear oil (193 mg, 18%).

¹ H NMR (300 MHz, CDCl ₃ ): 7.63 (s, 2H), 7.27 (m, 6H), 5.29 (s, 1H), 4.69 (t, 1H, J = 4.80), 3.41 (s, 6H), 3.27 (m, 2H), 3.08 (m, 4H), 2.03 (s, 1H)

MS (ESI ⁺ , m / z): 565 [M + H] ⁺

Step (3): Preparation of (S) -N1- (2,2-dimethoxyethyl) -N1-phenethyl-3-phenylpropane-1,2-diamine

The compound (190mg, 0.336mmol) obtained in step (2) was dissolved in dichloromethane (2mL), piperidine (0.2mL) was added thereto, and the mixture was stirred for one hour. The reaction was concentrated and purified by column chromatography to give the title compound as a clear oil (89 mg, 77%).

MS (ESI ⁺ , m / z): 343 [M + H] ⁺

Step (4): (S)-(9H-Floren-9-yl) methyl 2- (1-((2,2-dimethoxyethyl) (phenethyl) amino) -3-phenylpropan-2-yl Preparation of Amino) -2-oxoethylcarbamate

The compound (380 mg, 1.11 mmol) obtained in step (3) was dissolved in dichloromethane (3 mL), followed by Fmoc-Gly-OH (395 mg, 1.33 mmol), EDCI (255 mg, 1.33 mmol), and HOBT (204 mg, 1.33 mmol). ), And DIPEA (0.58 mL, 3.33 mmol) were dissolved in dichloromethane (3 mL) and stirred at room temperature for 16 hours. After the reaction, 5% citric acid was added dropwise to the solution to separate the organic layer, and the organic layer was dried over sodium sulfate, filtered under reduced pressure and distilled under reduced pressure. The obtained residue was purified by column chromatography to give the title compound as a white solid (590 mg, 86%).

¹ H NMR (300 MHz, CDCl ₃ ): 7.63 (m, 2H), 7.57 (m, 2H), 7.41 (m, 2H), 7.26 (m, 12H), 4.41 (m, 2H), 4.22 (m, 1H ), 4.14 (m, 1H), 3.85 (m, 1H), 3.27 (s, 2H), 4.22 (m, 1H), 4.14 (m, 1H), 3.85 (m, 1H), 3.27 (s, 2H) , 3.24 (d, 6H), 2.91 (m, 10H)

MS (ESI ⁺ , m / z): 622 [M + H] ⁺

Step (5): Preparation of (S) -2-amino-N- (1-((2,2-dimethoxyethyl) (phenethyl) amino) -3-phenylpropan-2-yl) acetamide

The compound (330 mg, 0.43 mmol) obtained in step (4) was dissolved in dichloromethane (10 mL), piperidine (1 mL) was added thereto, and the mixture was stirred for one hour. The reaction was concentrated and purified by column chromatography to give the title compound as a yellow oil (155 mg, 97%).

¹ H NMR (300MHz, CDCl ₃ ): 7.27-7.07 (m, 10H), 4.36 (t, 1H, J = 5.18), 4.14 (t, 1H, J = 6.83), 3.31 (s, 6H), 3.20 ( s, 2H), 2.87 (d, 2H, J = 6.41), 2.83 (m, 2H), 2.72 (m, 4H), 2.58 (m, 2H), 1.41 (s, 2H)

MS (ESI ⁺ , m / z): 400 [M + H] ⁺

Step (6): (9H-Floren-9-yl) methyl (7S, 13S) -7-benzyl-13-cyclohexyl-3-methoxy-9,12-dioxo-5-phenethyl-2- Preparation of oxa-5,8,11-triaztridecane-13-ylcarbamate

After dissolving the compound (155 mg, 0.388 mmol) obtained in the step (5) in dichloromethane (3 mL), Fmoc-Chg-OH (177 mg, 0.466 mmol), EDCI (89 mg, 0.466 mmol), HOBT (71 mg, 0.466 mmol) ), And DIPEA (0.2 mL, 1.164 mmol) were dissolved in dichloromethane (3 mL) and stirred at room temperature for 16 hours. After the reaction, 5% citric acid was added dropwise to the solution to separate the organic layer, and the organic layer was dried over sodium sulfate, filtered under reduced pressure and distilled under reduced pressure. The obtained residue was purified by column chromatography to give the title compound as a white solid (290 mg, 98%).

MS (ESI ⁺ , m / z): 761 [M + H] ⁺

Step (7): (9H-Floren-9-yl) methyl (S) -2-((5S, 8aR) -5-benzyl-3-oxo-7-phenethylhexahydroimidazo [1,2- a] Preparation of pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethylcarbamate

The compound (330 mg, 0.43 mmol) obtained in step (6) was dissolved in formic acid (10 mL) and stirred at 100 ° C. for 3 hours. The reaction was concentrated and purified by column chromatography to give the title compound as a yellow solid (85 mg, 28%).

¹ H NMR (300MHz, CDCl ₃ ): 8.25 (s, 1H), 7.70 (m, 2H), 7.57 (m, 2H), 7.28 (m, 14H), 5.62 (m, 1H), 4.25 (m, 5H ), 3.28 (m, 1H), 3.04 (m, 3H), 2.76 (m, 3H), 1.69 (m, 6H), 0.90 (m, 6H)

MS (ESI ⁺ , m / z): 697 [M + H] ⁺

Step (8): (5S, 8aR) -1-((S) -2-Amino-2-cyclohexylacetyl) -5-benzyl-7-phenethylhexahydroimidazo [1,2-a] pyrazine- Preparation of 3 (5H) -one

The compound (85 mg, 0.12 mmol) obtained in step (7) was dissolved in dichloromethane (10 mL), piperidine (1 mL) was added thereto, and the mixture was stirred for one hour. The reaction was concentrated and purified by column chromatography to give the title compound as a white solid (51 mg, 88%).

¹ H NMR (300MHz, CDCl ₃ ): 7.33 (m, 10H), 5.67 (m, 1H), 4.05 (m, 1H), 3.34 (m, 2H), 3.17 (m, 3H), 2.78 (m, 4H ), 2.12 (m, 3H), 1.88 (m, 7H), 1.26 (m, 8H), 0.92 (m, 1H)

MS (ESI ⁺ , m / z): 475 [M + H] ⁺

Step (9): t-butyl (S) -1-((S) -2-((5S, 8aR) -5-benzyl-3-oxo-7-phenethylhexahydroimidazo [1,2-a ] Preparation of pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethylamino) -1-oxopropan-2-yl (methyl) carbamate

Boc-MeAla-OH (26mg, 0.129mmol) was dissolved in MC (3mL), and EDCI (25mg, 0.129mmol), HOBT (20mg, 0.129mmol), and DIPEA (0.06mL, 0.32mmol) were added dropwise and at room temperature. Stir for 15 minutes. To the reaction solution was added dropwise the compound (51 mg, 0.108 mmol) obtained in step (8), and stirred at room temperature for 4 hours. After the reaction, sodium bicarbonate (10 mL) was added dropwise to the solution to separate the organic layer, and the organic layer was dried over sodium sulfate, filtered under reduced pressure and distilled under reduced pressure. The obtained residue was purified by column chromatography to give the title compound as a white solid (52 mg, 73%).

MS (ESI ⁺ , m / z): 660 [M + H] ⁺

Step (10): (S) -N-((S) -2-((5S, 8aR) -5-benzyl-3-oxo-7-phenethylhexahydroimidazo [1,2-a] pyrazine- Preparation of 1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide hydrochloride

The compound (52 mg, 73%) obtained in step (9) was dissolved in 4M HCl / dioxane solution (10 mL) and stirred at room temperature for 1.5 hours. After the reaction, the solution was concentrated under reduced pressure and recrystallized using MC and ethyl acetate to give the title compound as a white solid (35 mg, 74%).

MS (ESI ⁺ , m / z): 582 [M + Na] ⁺

Example 12: (S) -N-((S) -2-((5S, 8aR) -5-benzyl-3,6-dioxo-7- (3-phenylpropyl) hexahydroimidazo [1, 2-a] Pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide hydrochloride

The title compound was obtained as a white solid (37 mg, 4%) through the same method as in Example 1, except that 3-phenylpropylamine was used instead of the phenethylamine of Step 1 in Example 1.

MS (ESI ⁺ , m / z): 588 [M + H] ⁺

Example 13: (S) -N-((S) -2-((5S, 8aR) -5-benzyl-7- (4-fluorophenethyl) -3,6-dioxohexahydroimidazo [ 1,2-a] Pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide hydrochloride

The title compound was obtained as a white solid through the same method as Example 1 except for using 4-fluorophenethylamine in Example 1 instead of phenethylamine in Step 1 (6 mg, 14%).

¹ H NMR (300MHz, CD ₃ OD): 8.31 (d, 1H), 7.82-7.72 (m, 3H), 7.45-7.00 (m, 6H), 6.67-6.61 (m, 1H), 5.76 (s, 1H ), 5.49-5.39 (m, 2H), 4.96 (s, 1H), 4.60-4.51 (m, 1H), 4.36-3.73 (m, 5H), 3.56-3.41 (m, 1H), 3.26-3.09 (m , 2H), 2.68 (s, 3H), 1.78 (br, 6H), 1.52 (d, 3H), 1.28-0.94 (m, 5H)

MS (ESI ⁺ , m / z): 592 [M + H] ⁺

Example 14: (S) -N-((S) -3,3-dimethyl-1-((5S, 8aR) -5- (naphthalen-1-ylmethyl) -3,6-dioxo-7- Preparation of phenethylhexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-oxobutan-2-yl) -2- (methylamino) propanamide

The title compound was obtained as a white solid by the same method as Example 1 except for using Fmoc-3- (1-naphthyl) -alanine in Example 1 instead of Fmoc-Phe-OH in Step 2 (8 mg, 15 %).

MS (ESI ⁺ , m / z): 598 [M + H] ⁺

Example 15: (S) -N-((S) -1-cyclohexyl-2-((2S, 5S, 8aR) -2-methyl-5- (naphthalen-1-ylmethyl) -3,6- Preparation of Dioxo-7-phenethylhexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -2-oxoethyl) -2- (methylamino) propanamide

The same method as Example 1 was used in Example 1 except that Fmoc-2-Nal-OH was used instead of Fmoc-Phe-OH in Step 2 and Fmoc-Ala-OH was used instead of Fmoc-Gly-OH in Step 4. The title compound was obtained as a white solid (6 mg, 14%).

MS (ESI ⁺ , m / z): 638 [M + H] ⁺

Example 16: (S) -N-((S) -2-((5S, 8aR) -7-benzyl-5- (naphthalen-2-ylmethyl) -3,6-dioxohexahydroimidazo [ 1,2-a] Pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide

Except for using benzylamine in place of phenethylamine in Example 1, using Fmoc-2-Nal-OH instead of Fmoc-Phe-OH in step 2, and formic acid instead of 4M HCl in step 10 The title compound was obtained in the same manner as in Example 1 as a white solid (50 mg, 34%).

¹ H NMR (300MHz, CD ₃ OD): 8.67 (br, 1H), 7.87-7.78 (m, 3H), 7.43-7.35 (m, 2H), 7.28 (d, 1H), 6.86 (t, 1H), 6.75 (t, 2H), 6.50-6.47 (m, 3H), 6.07 (br, 1H), 5.43 (d, 1H), 5.06 (d, 1H), 4.93 (s, 1H), 4.40-4.30 (m, 3H), 4.20-3.97 (m, 3H), 3.80 (m, 1H), 3.41-3.38 (m, 2H), 3.32-3.29 (m, 2H), 2.59 (s, 3H), 1.75-1.62 (m, 6H), 1.45 (t, 3H), 1.28-1.09 (m, 5H)

MS (ESI ⁺ , m / z): 610 [M + H] ⁺

Example 17: (S) -N-((S) -2-((5S, 8aR) -7- (4-chlorobenzyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxo Preparation of Hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide

In the same manner as in Example 16, except that 4-chlorobenzylamine was used instead of the benzylamine of Example 1, the title compound was obtained as a white solid (54 mg, 35%).

¹ H NMR (300MHz, CD ₃ OD): 8.47 (br, 1H), 7.88-7.81 (m, 3H), 7.48-7.43 (m, 3H), 7.29 (d, 1H), 6.73-6.69 (m, 2H ), 6.49-6.42 (m, 3H), 6.06 (br, 1H), 6.06 (br, 1H), 5.41 (d, 1H), 5.05 (d, 1H), 4.96 (br, 1H), 4.91 (br, 1H), 4.38-4.08 (m, 4H), 3.94-3.81 (m, 3H), 3.-45-3.38 (m, 1H), 3.29-3.26 (m, 1H), 2.54 (s, 3H), 1.77 -1.65 (m, 6H), 1.46 (d, 3H), 1.28-0.89 (m, 5H)

MS (ESI ⁺ , m / z): 644 [M + H] ⁺

Example 18: (S) -N-((S) -2-((5S, 8aR) -7- (4-methoxybenzyl) -5- (naphthalen-2-ylmethyl) -3,6-di Preparation of oxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide

The title compound was obtained as a white solid (40 mg, 30%) through the same method as Example 16 except for using 4-methoxybenzylamine instead of the benzylamine of Step 1 in Example 16.

¹ H NMR (300MHz, CD ₃ OD): 8.43 (br, 1H), 7.73-7.64 (m, 3H), 7.32-7.31 (m, 2H), 7.14 (d, 1H), 6.36-6.29 (m, 3H ), 6.17-6.14 (m, 2H), 5.93 (br, 1H), 5.29 (d, 1H), 4.93 (d, 1H), 4.72 (d, 1H), 4.30-3.65 (m, 6H), 3.33- 3.25 (m, 2H), 3.44 (s, 3H), 3.17-3.09 (m, 1H), 2.48 (s, 3H), 1.63-1.51 (m, 6H), 1.34 (br, 3H), 1.17-0.98 ( m, 5H)

MS (ESI ⁺ , m / z): 640 [M + H] ⁺

Example 19: (S) -N-((S) -2-((5S, 8aR) -7- (4-methoxyphenethyl) -5- (naphthalen-2-ylmethyl) -3,6- Preparation of Dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide

In the same manner as in Example 16 except that 4-methoxyphenethylamine was used instead of the benzylamine of Step 1 in Example 16, the title compound was obtained as a white solid (42 mg, 31%).

¹ H NMR (300MHz, CD ₃ OD): 8.56 (br, 1H), 7.92-7.80 (m, 3H), 7.55-7.50 (m, 2H), 7.23 (d, 1H), 6.49-6.43 (m, 3H ), 6.30-6.24 (m, 2H) 5.93 (br, 1H), 5.33 (d, 1H), 4.42-3.95 (m, 4H), 3.83-3.70 (m, 3H), 3.60 (s, 3H), 3.28 -3.07 (m, 3H), 2.60 (s, 3H), 2.52-2.47 (m, 2H), 1.75-1.62 (m, 6H), 1.45 (d, 3H), 1.29-1.10 (m, 6H)

MS (ESI ⁺ , m / z): 654 [M + H] ⁺

Example 20: (S) -N-((S) -2-((5S, 8aR) -7- (4-fluorophenethyl) -5- (naphthalen-2-ylmethyl) -3,6- Preparation of Dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide

In Example 1, 4-fluorophenethylamine was used instead of phenethylamine in Step 1, Fmoc-1-Nal-OH was used instead of Fmoc-Phe-OH in Step 2, and formic acid was used instead of 4M HCl in Step 10. The title compound was obtained as a white solid (30 mg, 6%) through the same method as in Example 1 except using.

¹ H NMR (300 MHz, CDCl ₃ ): 8.00-7.87 (m, 1H), 7.79-7.76 (m, 1H), 7.71-7.68 (m, 1H), 7.54-7.50 (m, 2H), 7.20-7.18 ( m, 1H), 7.15-7.05 (br, 1H), 6.56-6.50 (m, 2H), 6.42-6.37 (m, 2H), 5.60-5.55 (br, 1H), 5.29 (d, J = 5.7 Hz, 1H), 4.80 (d, J = 5.7 Hz, 1H), 4.35-4.32 (m, 2H), 4.21-4.16 (m, 1H), 4.10-4.00 (m, 1H), 3.82-3.80 (m, 2H) , 3.31-3.23 (m, 2H), 3.11-3.07 (m, 2H), 2.58-2.56 (m, 2H), 2.39 (s, 3H), 1.72-1.61 (m, 6H), 1.29 (s, 3H) , 1.28-1.02 (m, 4H)

MS (ESI ⁺ , m / z): 642 [M + H] ⁺

Example 21: (S) -N-((S) -2-((5S, 8aR) -7- (4-fluorophenethyl) -5- (naphthalen-2-ylmethyl) -3,6- Preparation of Dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide

In Example 1 4-fluorophenethylamine is used instead of phenethylamine in Step 1, Fmoc-1-Nal-OH is used instead of Fmoc-Phe-OH in Step 2, and Fmoc-Chg-OH in Step 6 The title compound was obtained as a white solid through the same method as Example 1 except using Fmoc-Tle-OH instead of formic acid instead of 4M HCl in step 10 (24 mg, 6%).

¹ H NMR (300 MHz, CDCl ₃ ): 7.90-7.76 (m, 3H), 7.74-7.70 (m, 1H), 7.54-7.50 (m, 2H), 7.20-7.18 (m, 1H), 7.00-6.75 ( br, 1H), 6.56-6.51 (m, 2H), 6.43-6.37 (m, 2H), 5.55 (m, 1H), 5.48 (d, J = 5.7 Hz, 1H), 4.75 (d, J = 5.7 Hz , 1H), 4.33-4.30 (m, 2H), 4.20-4.00 (m, 1H), 3.83-3.81 (m, 2H), 3.75-3.60 (m, 1H), 3.32-3.19 (m, 2H), 3.12 -3.06 (m, 2H), 2.58-2.56 (m, 2H), 2.37 (s, 3H), 1.30 (s, 3H), 1.02 (s, 9H)

MS (ESI ⁺ , m / z): 616 [M + H] ⁺

Example 22: (S) -N-((S) -2-((5S, 8aR) -7- (4-fluorophenethyl) -5- (4-methylbenzyl) -3,6-dioxo Preparation of Hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide

In Example 1, 4-fluorophenethylamine was used instead of phenethylamine in Step 1, Fmoc-Phe (4-Me) -OH instead of Fmoc-Phe-OH in Step 2, and 4M HCl in Step 10 The title compound was obtained as a white solid (20 mg, 4%) by the same method as Example 1 except for using formic acid instead.

¹ H NMR (300 MHz, CDCl ₃ ): 7.69 (d, J = 9.0 Hz, 1H), 7.11-6.99 (m, 4H), 6.77-6.72 (m, 4H), 5.70 (d, J = 3.6 Hz, 1H ), 5.35 (d, J = 5.7 Hz, 1H), 4.89 (brs, 1H), 4.60 (brs, 1H), 4.34-4.04 (m, 3H), 3.74-3.70 (m, 2H), 3.16-3.04 ( m, 4H), 2.64-2.63 (m, 2H), 2.41 (d, J = 8.1 Hz, 3H), 2.33 (s, 3H), 1.82-1.68 (m, 6H), 1.29 (s, 3H), 1.25 -0.98 (m, 4H)

MS (ESI ⁺ , m / z): 606 [M + H] ⁺

Example 23: (S) -N-((S) -2-((5S, 8aR) -7- (4-fluorophenethyl) -5- (4-methylbenzyl) -3,6-dioxo Preparation of hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide

In Example 1 4-fluorophenethylamine is used instead of phenethylamine in Step 1, Fmoc-Phe (4-Me) -OH is used in place of Fmoc-Phe-OH in Step 2, and Fmoc- in Step 6 is used. The title compound was obtained as a white solid by the same method as Example 1 except for using Fmoc-Tle-OH instead of Chg-OH and formic acid instead of 4M HCl in step 10 (15 mg, 4%).

¹ H NMR (300MHz, CDCl ₃ ): 7.82-7.79 (m, 1H), 7.04-6.70 (m, 8H), 5.70 (brs, 1H), 5.30 (d, J = 5.7 Hz, 1H), 4.85 (brs , 1H), 4.60 (brs, 1H), 4.30-4.24 (m, 1H), 4.04 (d, J = 4.2 Hz, 1H), 4.00-3.75 (m, 1H), 3.72-3.61 (m, 2H), 3.11-2.98 (m, 4H), 2.60 (m, 2H), 2.35 (s, 3H), 2.28 (s, 3H), 1.26 (d, J = 6.9 Hz, 3H), 1.21 (s, 9H)

MS (ESI ⁺ , m / z): 580 [M + H] ⁺

Example 24: (S) -N-((S) -2-((5S, 8aR) -7- (3-chlorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahexa Preparation of Hydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide

Example 16 The same procedure as Example 16 except for using 3-chlorophenethylamine instead of benzylamine of Step 1 and Fmoc-Phe (4-Me) -OH instead of Fmoc-Phe-OH in Step 2. Through the method the title compound was obtained as a white solid (20 mg, 30%).

¹ H NMR (300MHz, CD ₃ OD): 8.51 (s, 1H), 7.26-7.02 (m, 7H), 6.70 (t, 1H), 5.70 (d, 1H), 5.34 (d, 1H), 5.21 ( d, 1H), 4.79 (d, 1H), 4.36-4.11 (m, 3H), 3.94-3.71 (m, 3H), 3.13-2.82 (m, 2H), 2.66-2.62 (m, 5H), 2.34 ( d, 3H), 1.76-7.69 (m, 6H), 1.48 (d, 3H), 1.34-1.08 (m, 5H)

MS (ESI ⁺ , m / z): 622 [M + H] ⁺

Example 25: (S) -N-((S) -2-((5S, 8aR) -7- (2-chlorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexa Preparation of Hydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide

Same as Example 16 except for using 2-chlorophenethylamine instead of benzylamine of Step 1 in Example 16 and Fmoc-Phe (4-Me) -OH instead of Fmoc-Phe-OH in Step 2 The title compound was obtained as a white solid (23 mg, 32%) by the method.

¹ H NMR (300MHz, CD ₃ OD): 8.50 (s, 1H), 7.38-6.94 (m, 7H), 6.71 (t, 1H), 5.68 (br, 1H), 5.33 (br, 1H), 5.21 ( d, 1H), 4.37-3.61 (m, 7H), 3.40-3.34 (m, 1H), 3.26-2.94 (m, 3H), 2.80 (t, 3H), 1.79-1.69 (m, 6H), 1.49 ( d, 3H), 1.30-0.98 (m, 6)

MS (ESI ⁺ , m / z): 622 [M + H] ⁺

Example 26: (S) -N-((S) -2-((5S, 8aR) -5,7-bis (4-methylbenzyl) -3,6-dioxohexahydroimidazo [1,2 Preparation of pyrazin-1 (5H) -yl-3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide

Example 6 uses 4-methylbenzylamine instead of benzylamine, step 2 uses Fmoc-Phe (4-Me) -OH instead of Fmoc-Phe-OH and step 6 Fmoc-Chg-OH The title compound was obtained as a white solid by the same method as Example 16 except for using Fmoc-Tle-OH instead (21 mg, 28%).

¹ H NMR (300MHz, CD ₃ OD): 7.73-7.67 (m, 2H), 7.32-7.25 (m, 2H), 7.12-7.09 (m, 2H), 6.89-6.85 (m, 2H), 6.54-6.52 (m, 2H), 5.71 (br, 1H), 5.38 (br, 1H), 5.31 (d, 1H), 5.04-4.91 (m, 2H), 4.39-3.91 (m, 5H), 3.76-3.61 (m , 2H), 3.24-3.14 (m, 2H), 2.56 (d, 3H), 2.24 (s, 6H), 1.44-1.41 (m, 3H), 1.04 (d, 9H)

MS (ESI ⁺ , m / z): 562 [M + H] ⁺

Example 27: (S) -N-((S) -1-((5S, 8aR) -7- (3-chlorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahexa Preparation of Hydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide

In the same manner as in Example 26, except that 3-chlorophenethylamine was used instead of 4-methylbenzylamine of Example 1, the title compound was obtained as a white solid (15 mg, 24%).

¹ H NMR (300MHz, CD ₃ OD): 7.31-6.98 (m, 8H), 5.69 (br, 1H) 5.33 (br, 1H), 5.21 (d, 1H), 4.81 (d, 1H), 4.43-3.71 (m, 6H), 3.27-2.80 (m, 4H), 2.66-2.64 (m, 2H), 2.33 (s, 6H), 1.29 (d, 3H), 1.05-0.99 (m, 12H)

MS (ESI ⁺ , m / z): 596 [M + H] ⁺

Example 28: (S) -N-((S) -2-((5S, 8aR) -7- (4-pyridinemethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxo Preparation of Hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide

In Example 1, 4-pyridinemethylamine is used instead of phenethylamine in Step 1, Fmoc-1-Nal-OH is used instead of Fmoc-Phe-OH in Step 2, and formic acid is used instead of 4M HCl in Step 10. The title compound was obtained as a white solid (12 mg, 3%) through the same method as in Example 1.

MS (ESI ⁺ , m / z): 611 [M + H] ⁺

Example 29: (S) -N-((S) -2-((5S, 8aR) -7- (4-pyridinemethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxo Preparation of hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide

4-pyridinemethylamine instead of phenethylamine in Step 1, Fmoc-1-Nal-OH instead of Fmoc-Phe-OH in Step 2, Fmoc instead of Fmoc-Chg-OH in Step 6 The title compound was obtained as a white solid through the same method as Example 1 except using -Tle-OH and using formic acid instead of 4M HCl in step 10 (30 mg, 7%).

MS (ESI ⁺ , m / z): 585 [M + H] ⁺

Example 30: (S) -N-((S) -2-((5S, 8aR) -7- (methyl- (4-methylbenzoate))-5- (naphthalen-2-ylmethyl) -3 Preparation of, 6-dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide

In Example 1, methyl- (4-aminomethyl) benzoate was used instead of phenethylamine in Step 1, Fmoc-1-Nal-OH was used instead of Fmoc-Phe-OH in Step 2, and 4M HCl in Step 10 was used. The title compound was obtained as a white solid (16 mg, 3%) through the same method as Example 1 except using formic acid.

MS (ESI ⁺ , m / z): 668 [M + H] ⁺

Example 31: (S) -N-((S) -2-((5S, 8aR) -7- (methyl- (4-methylbenzoate))-5- (naphthalen-2-ylmethyl) -3 , 6-dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide Manufacture

In Example 1, methyl- (4-aminomethyl) benzoate is used instead of phenethylamine in Step 1, Fmoc-1-Nal-OH is used instead of Fmoc-Phe-OH in Step 2, and Fmoc-Chg in Step 6 The title compound was obtained as a white solid by the same method as Example 1 except for using Fmoc-Tle-OH instead of -OH and formic acid instead of 4M HCl in step 10 (18 mg, 4%).

MS (ESI ⁺ , m / z): 642 [M + H] ⁺

Example 32: (S) -N-((S) -1-((5S, 8aR) -7- (2-chlorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexa Preparation of Hydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide

In the same manner as in Example 26, except that 2-chlorophenethylamine was used instead of 4-methylbenzylamine of Step 1, the title compound was obtained as a white solid (15 mg, 24%).

¹ H NMR (300MHz, CD ₃ OD): 7.29-6.94 (m, 7H), 6.71 (t, 1H), 5.67 (br, 1H), 5.33 (br, 1H), 5.20 (d, 1H), 4.79 ( d, 1H), 4.42-3.81 (m, 4H), 3.78-3.71 (m, 2H), 3.42-2.94 (m, 5H), 2.81 (t, 2H), 2.35-2.32 (m, 6H), 1.26 ( d, 3H), 1.05-0.92 (m, 9H)

MS (ESI ⁺ , m / z): 596 [M + H] ⁺

Example 33: (S) -N-((S) -3,3-dimethyl-1-((5S, 8aR) -5- (naphthalen-2-ylmethyl) -3,6-dioxo-7- Preparation of phenethylhexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-oxobutan-2-yl) -2- (methylamino) propanamide

Example 26 except for using phenethylamine instead of 4-methylbenzylamine in Step 1 and Fmoc-2-Nal-OH instead of Fmoc-Phe (4-Me) -OH in Step 2 Through the same method as the title compound was obtained as a white solid (22 mg, 4%).

¹ H NMR (300MHz, CD ₃ OD): 8.54 (s, 1H), 7.94-7.83 (m, 3H), 7.57-7.52 (m, 2H), 7.27 (d, 1H), 6.76-6.59 (m, 5H ), 6.42 (d, 1H), 5.97 (d, 1H), 5.33 (d, 1H), 4.93 (d, 1H), 4.42-4.29 (m, 2H), 4.15-4.08 (m, 1H), 3.83- 3.71 (m, 3H), 3.29-3.14 (m, 3H), 2.56-2.53 (m, 5H), 1.41 (t, 3H), 1.06-0.91 (m, 9H)

MS (ESI ⁺ , m / z): 598 [M + H] ⁺

Example 34: (S) -N-((S) -2-((5S, 8aR) -7- (4-chlorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahexa Preparation of Hydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide

In the same manner as in Example 1 except for using 4-chlorophenethylamine instead of 4-methylbenzylamine in Step 26 and Fmoc-Chg instead of Fmoc-Tle-OH in Step 6 The compound was obtained as a white solid (24 mg, 4%).

¹ H NMR (300MHz, CD ₃ OD): 8.49 (s, 1H), 7.74-7.67 (m, 4H), 7.34-7.28 (m, 4H), 7.06-7.01 (m, 6H), 6.79-6.77 (m , 2H), 5.68 (br, 1H), 5.31 (br, 1H), 5.19 (d, 1H), 4.79 (br, 1H), 4.33-3.80 (m, 8H), 3.24-2.82 (m, 5H), 2.64 (br, 5H), 2.36 (d, 3H), 1.78-1.75 (m, 6H), 1.48 (d, 3H), 1.35-1.05 (m, 6H)

MS (ESI ⁺ , m / z): 622 [M + H] ⁺

Example 35: (S) -N-((S) -1-((5S, 8aR) -7- (4-chlorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexa Preparation of Hydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide

The title compound was obtained as a white solid (20 mg, 4%) in the same manner as in Example 1 except that 4-chlorophenethylamine was used instead of 4-methylbenzylamine of Step 1 in Example 26.

¹ H NMR (300MHz, CD ₃ OD): 8.39 (br, 1H), 7.19-6.90 (m, 6H), 6.97 (d, 2H), 5.58 (br, 1H), 5.20 (br, 1H), 5.08 ( d, 1H), 4.69 (d, 1H), 4.19-3.69 (m, 8H), 3.10-2.72 (m, 4H), 2.56-2.49 (m, 5H), 2.25 (d, 3H), 1.35-1.29 ( m, 3H), 0.95-0.87 (m, 9H)

MS (ESI ⁺ , m / z): 596 [M + H] ⁺

Example 36 methyl 4-(((5S) -5- (biphenyl-4-ylmethyl) -1-((S) -2-cyclohexyl-2-((S) -2- (methylamino) Preparation of propaneamido) acetyl) -3,6-dioxohexahydroimidazo [1,2-a] pyrazin-7 (1H) -yl) methyl) benzoate

The title compound was obtained as a white solid (16 mg, 3%) in the same manner as in Example 1 except for using Fmoc-Bip-OH instead of Fmoc-Phe-OH in Example 30.

MS (ESI ⁺ , m / z): 594 [M + H] ⁺

Example 37: (S) -N-((S) -2-((5S, 8aR) -7- (3-chlorophenethyl) -5- (naphthalen-2-ylmethyl) -3,6-di Preparation of oxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide

In Example 1 3-chlorophenethylamine was used instead of phenethylamine in Step 1, Fmoc-1-Nal-OH was used instead of Fmoc-Phe-OH in Step 2, and Fmoc-Chg-OH in Step 6 was used. The title compound was obtained as a white solid (83 mg, 9%) using the same method as Example 1 except using Fmoc-Tle-OH and using formic acid instead of 4M HCl in step 10.

MS (ESI ⁺ , m / z): 633 [M + H] ⁺

Example 38: (S) -N-((S) -2-((5S, 8aR) -7- (2-chlorophenethyl) -5- (naphthalen-2-ylmethyl) -3,6-di Preparation of oxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide

In Example 1 2-chlorophenethylamine is used instead of phenethylamine in Step 1, Fmoc-1-Nal-OH is used instead of Fmoc-Phe-OH in Step 2, and instead of Fmoc-Chg-OH in Step 6 The title compound was obtained as a white solid (61 mg, 6%) by the same method as Example 1 except using Fmoc-Tle-OH and using formic acid instead of 4M HCl in step 10.

MS (ESI ⁺ , m / z): 633 [M + H] ⁺

Example 39: (S) -N-((S) -2-((5S, 8aR) -7- (2-phenylpropyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxo Preparation of hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide

In Example 1 2-phenylpropylamine was used instead of phenethylamine in Step 1, Fmoc-1-Nal-OH was used in place of Fmoc-Phe-OH in Step 2, and Fmoc instead of Fmoc-Chg-OH in Step 6 The title compound was obtained as a white solid (113 mg, 12%) by the same method as Example 1 except using -Tle-OH and using formic acid instead of 4M HCl in step 10.

MS (ESI ⁺ , m / z): 612 [M + H] ⁺

Example 40: (2S) -N-((2S) -1-((5S) -5-((1H-indol-3-yl) methyl) -3,6-dioxo-7-pentylhexahydroimide Preparation of Dazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl-2- (methylamino) propanamide

Example 1 uses Fmoc-tryptophan instead of Fmoc-Phe-OH in step 2, except using Fmoc-Tle-OH instead of Fmoc-Chg-OH in step 6 and formic acid instead of 4M HCl in step 10 The title compound was obtained as a white solid through the same method as in Example 1 (21 mg, 3%).

MS (ESI ⁺ , m / z): 587 [M + H] ⁺

Example 41: (2S) -N-((1S) -2-((5S) -5-((1H-indol-3-yl) methyl) -3,6-dioxo-7-pentylhexahydroimide Preparation of Dazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide

The title compound was obtained as a white solid in the same manner as in Example 1 except for using Fmoc-tryptophan instead of Fmoc-Phe-OH in Example 1 and formic acid instead of 4M HCl in Step 10 ( 17 mg, 2%).

MS (ESI ⁺ , m / z): 613 [M + H] ⁺

Example 42: (S) -N-((S) -2-((5S, 8aR) -7- (2-fluorophenethyl) -5- (naphthalen-2-ylmethyl) -3,6- Preparation of Dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide

In Example 1 2-fluorophenethylamine was used instead of phenethylamine in Step 1, Fmoc-1-Nal-OH was used instead of Fmoc-Phe-OH in Step 2, and Fmoc-Chg-OH in Step 6 Fmoc-Tle-OH was used instead, and the title compound was obtained as a white solid through the same method as Example 1 except using formic acid instead of 4M HCl in step 10 (17 mg, 2%).

MS (ESI ⁺ , m / z): 616 [M + H] ⁺

Example 43: (S) -N-((S) -2-((5S, 8aR) -7- (2-bromophenethyl) -5- (naphthalen-2-ylmethyl) -3,6-di Preparation of oxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide

In Example 1 2-bromophenethylamine was used instead of phenethylamine in Step 1, Fmoc-1-Nal-OH was used instead of Fmoc-Phe-OH in Step 2, and instead of Fmoc-Chg-OH in Step 6 The title compound was obtained as a white solid (32 mg, 3%) by the same method as Example 1 except using Fmoc-Tle-OH and using formic acid instead of 4M HCl in step 10.

MS (ESI ⁺ , m / z): 677 [M + H] ⁺

Example 44: (S) -N-((S) -2-((5S, 8aR) -7- (2-methoxyphenethyl) -5- (naphthalen-2-ylmethyl) -3,6- Preparation of Dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide

In Example 1 2-methoxyphenethylamine was used instead of phenethylamine in Step 1, Fmoc-1-Nal-OH instead of Fmoc-Phe-OH in Step 2, and Fmoc-Chg-OH in Step 6 Fmoc-Tle-OH was used instead, and the title compound was obtained as a white solid through the same method as Example 1 except using formic acid instead of 4M HCl in step 10 (28 mg, 3%).

MS (ESI ⁺ , m / z): 628 [M + H] ⁺

Example 45: (S) -N-((S) -2-((5S, 8aR) -7- (2,3-dihydro-1H-inden-2-yl) -5- (naphthalene-2- Ylmethyl) -3,6-dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- ( Preparation of Methylamino) propanamide

In Example 1 2,3-dihydro-1H-inden-2-amine was used instead of phenethylamine in Step 1, Fmoc-1-Nal-OH was used in place of Fmoc-Phe-OH in Step 2, and The title compound was obtained as a white solid through the same method as Example 1 except for using Fmoc-Tle-OH instead of Fmoc-Chg-OH of 6 and formic acid instead of 4M HCl in Step 10 (23 mg, 3 %).

MS (ESI ⁺ , m / z): 610 [M + H] ⁺

Example 46: (S) -N-((S) -2-((5S, 8aR) -7- (4-methyl-1,2,3,4-tetrahydronaphthalen-2-yl) -5- (Naphthalen-2-ylmethyl) -3,6-dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl Preparation of) -2- (methylamino) propanamide

In Example 1 4-methyl-1,2,3,4-tetrahydronaphthalen-2-amine was used instead of phenethylamine in Step 1, and Fmoc-1-Nal-OH instead of Fmoc-Phe-OH in Step 2 To obtain the title compound as a white solid in the same manner as in Example 1 except for using Fmoc-Tle-OH instead of Fmoc-Chg-OH in Step 6 and formic acid instead of 4M HCl in Step 10 (31 mg, 3%).

MS (ESI ⁺ , m / z): 638 [M + H] ⁺

Example 47: (S) -N-((S) -2-((5S, 8aR) -7- (1,2,3,4-tetrahydronaphthalen-2-yl) -5- (naphthalene-2 -Ylmethyl) -3,6-dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- Preparation of (methylamino) propanamide

Example 1 uses 1,2,3,4-tetrahydronaphthalen-2-amine in place of phenethylamine and Fmoc-1-Nal-OH in place of Fmoc-Phe-OH in step 2, The title compound was obtained as a white solid through the same method as Example 1 except for using Fmoc-Tle-OH instead of Fmoc-Chg-OH in Step 6 and formic acid instead of 4M HCl in Step 10 (28 mg, 3%).

MS (ESI ⁺ , m / z): 624 [M + H] ⁺

The structural formulas of the compounds obtained in Examples 1 to 47 are shown in Table 1 below.

Test Example

The bioassay test was performed on the compounds prepared in the above Examples.

Test Example 1 Evaluation of Caspase Activity

MDA-MB-231 breast cancer cells (ATCC # HTB-26) were seeded in a 96 well plate at a density of 1.0-1.5 × 10 ⁴ cells / well and treated with drugs 24 hours later.

The drug was treated at 0 μM, 0.5 μM, 1 μM, and 5 μM concentrations for 12 hours for caspase 3/7 and 24 hours for caspase 9, then the media was removed and MDA-MB-231 breast cancer cells (ATCC # HTB-26 ) Was washed twice with PBS at 4 ° C. and used for the experiment. Promega's Caspase-Glo ^TM 9 assay kit (Cat # .G8210) and Caspase-Glo ^™ 3/7 assay kit (Cat #. G8090) Was added to the cell sample at 100 μl / well. It was reacted for 2 hours at 37 ℃ using the following Tecan (Tecan)'s Infinite ^TM M1000 multi-reader (Infinite ^TM M1000 multi reader) was measured for light emission (luminescence).

The measurement values of the representative compounds are shown in Table 2 below, and Ebot's IAP antagonist was used as a control.

MDA-MB-231 breast cancer cell line, more than 6 times more than "***", less than 6 times more than 4 times "**", less than 3 times more than 1 times when the magnification compared to normal reference before drug treatment Are marked with "*".

Example Cascade-3 activation Example Cascade-3 activation standard One 10 *** contrast ** 32 *** 7 ** 37 *** 8 ** 38 *** 9 ** 44 **

Test Example 2 Evaluation of Affinity

In order to analyze the affinity with the BIR-3 domain, 4 μL of XIAP BIR diluted at 1.25 μM in black round-bottom 96-well plates were diluted to 0.0625 μM and then diluted to 4F (AbuRPF-K). 5-Fam) -NH 2) was added at 10 μl / well with light blocking. Here, XIAR BIR3 is the 241-356th amino acid residue of the human XIAP protein, and the pET28a vector (novagen) is subjected to standard DNA cloning procedures and PCR methods (Sambrook & Russell., Molecular cloning., Chapter 1. Third edition). Recombinant vector prepared using the above-described reference) was produced by transforming E. coli BL21 (DE3) cells.

Specifically, 1 ul of cDNA of K562 (human blood lympoblast-like leukemia) was used as a template, 1.5 mM MgCl ₂ , 0.2 mM dNPTs, 0.4 mM forward and reverse PCR primers (SEQ ID NOs: 1 and 2) and 2.0 unit tag polymerase. It was amplified by PCR using a reaction mixture containing (Elpis biotech, Inc., Korea). The PCR denatures the reaction mixture for 5 minutes, then repeats the reaction mixture for 35 cycles of 30 seconds at 94 ° C., 30 seconds at 52 ° C., 30 seconds at 72 ° C., and then extends at 72 ° C. for 7 minutes. (elongation).

DNA of the amplified XIAP BIR3 domain was inserted into pET28a vector (Novagen), which was transformed into E. coli BL21 (DE3) cells, and cultured. When the culture medium had an O.D value of 0.6, 0.1 mM IPTG was added to overexpress human XIAP BIR3 protein (SEQ ID NO: 3). The resulting protein terminus is capable of binding six histidines (His-tag).

Subsequently, the cells containing the overexpressed protein were crushed by an ultrasonic crusher and then centrifuged by a centrifuge to obtain only the supernatant. The supernatant was reacted with an affinity bead capable of specifically binding to His-tag and separated from other proteins, followed by use of 20 mM Tris, 10 mM NaCl (pH8.0), and 100 mM imidazole buffer, XIAP BIR3 protein was eluted from.

The binding assay buffer composed of 100 mM potassium phosphate (pH 7.5), 100 µg / ml bovine γ-globulin and 0.02% sodium azide was added at 105 µl / well and reacted at room temperature for 15 minutes. Subsequently, the positive control RPF-NH ₂ (alanine-arginine-proline-phenylalanine-NH ₂ ) and the test drug were sequentially diluted at a concentration of 0.01 to 1000 μM and added to 5 μl / well. DMSO was added instead to the 0% and 100% inhibition controls. After reacting for 3 hours with the light blocked, each measured by measuring the fluorescence polarization at the excitation wavelength of 485 nm and emission wavelength of 530 nm using Tecan's Infinite M1000 multireader. Ki values of each drug calculated using the IC ₅₀ value of the drug and the Kd value of 4F (ARPF-NH ₂ ) are shown in Table 3 below.

In the following Table 3, when the obtained Ki value is less than 100 nM, "***", when 100 nM or more and less than 1,000 nM, "**" is expressed as "*" when 1,000 nM or more.

Example BIR-3 domain affinity Example BIR-3 domain affinity One ** 38 ** 2 * 44 ** 32 ** 45 * 33 ** 46 ** 37 ** 47 **

Test Example 3 Evaluation of Cell Growth Inhibition ( in-vitro )

MDA-MB-231 breast cancer cells (ATCC # HTB-26) and BxPC-3 pancreatic cancer cells (ATCC #CRL 1687) reported to overexpress XIAP were purchased from American type culture collection (ATCC) Rock Bank, MD It was used for the experiment. MDA-MB-231 cell lines were matured in T-75 cm ² growth flasks under L-15 medium conditions containing 10% FBS and 1% penicillin / straptomycin (Gibco BRL). BxPC-3 cell lines were also grown in RPMI medium with 10% FBS and 1% penicillin / straptomycin. Fibroblasts Hs27 (ATCC # CRL 1634) and Balb / c3t3 (ATCC # CCL 163) were used for toxicological studies on normal cells. Several cell lines grown in growth media were transferred to 96 well culture plates at a density of 3,000 to 5,000 cells / 100 μl and incubated for 24 hours at 37 ° C., 5% CO ₂ , 95% air, and 100% relative humidity. (MDA-MB-231 cells were incubated at atmospheric conditions) Here, the test material was treated at a concentration of 10 μM to 0.1 nM, and MDA-MB-231 cell line was 120 hours, BxPC-3 was 96 hours, and normal cell line was 72 hours. Incubated.

MDA-MB-231 cell viability of cell lines Celta data ^{96 TM (CellTiter 96 AQueous One Solution} , MTS, Promega, Inc.) a was used to measure at 490nm with the absorption, BxPC-3 and cell viability of normal cells are 10% TCA The cells were fixed with (trichloroacetic acid) and stained with SRB (sulforhodamine B) solution to measure absorbance at 540 nm.

This yielded a GI ₅₀ value, a concentration at which the drug reduced cancer cell growth by 50%. Cancer cell growth rate was calculated by the following equation (1) or (2).

[Equation 1]

[(Ti-Tz) / (C-Tz)] x 100 (when Ti> = Tz)

[Equation 2]

[(Ti-Tz) / Tz] x 100 (for Ti <Tz)

In Equations 1 and 2, Tz is the absorbance in the 0% cell growth group, the cell density immediately before the test drug treatment, and C is the absorbance in the 100% cell growth group. Ti is the cell density of the test drug.

GI ₅₀ value is the test drug treatment concentration when the value of Equation 1 is 50, which means a concentration that inhibits cancer cell growth by 50%. In each measurement, comparison and equalization of each cell line were performed.

Activity values (GI ₅₀ values) for each compound are shown in Table 4 below, and Evot's IAP antagonist was used as a control.

In the activity measurement for MDA-MB-231 breast cancer cell line, "***" for less than 100nM, "**" for more than 100nM less than 1,000nM, "*" for more than 1,000nM. In addition, the normal cell lines Balb / C and Hs27 were expressed as "+++" when 10,000 nM or more, "++" when less than 10,000 nM, and more than 5,000 nM, and "+" when 5,000 nM or less.

Example MDA-
MB-231 Balb / c Hs27 Example MDA-
MB-231 Balb / c Hs27 contrast 230nM > 10,000nM > 10,000nM 24 ** +++ +++ One ** +++ +++ 25 ** +++ +++ 2 * +++ +++ 26 * +++ +++ 3 * +++ +++ 27 ** +++ +++ 4 * +++ +++ 28 * ++ +++ 5 * +++ +++ 29 * ++ +++ 6 * +++ +++ 30 * ++ +++ 7 ** ++ +++ 31 ** ++ +++ 8 ** +++ +++ 32 *** +++ +++ 9 ** +++ +++ 33 *** +++ +++ 10 ** +++ +++ 34 * +++ +++ 11 * +++ +++ 35 * +++ +++ 12 * +++ +++ 36 ** +++ +++ 13 ** +++ +++ 37 *** ++ +++ 14 ** ++ +++ 38 *** ++ +++ 15 * ++ +++ 39 ** ++ +++ 16 * ++ +++ 40 ** +++ +++ 17 * ++ +++ 41 ** +++ +++ 18 * ++ +++ 42 ** ++ +++ 19 * ++ +++ 43 ** ++ +++ 20 * ++ +++ 44 *** ++ +++ 21 ** ++ +++ 45 * ++ +++ 22 ** +++ +++ 46 ** ++ +++ 23 ** +++ +++ 47 ** ++ +++

As shown in Table 4, the compounds of Formula 1 of the present invention inhibited the growth of the MDA-MB-231 cell line overexpressing IAP very effectively at low drug concentration, and also as shown in Table 2, cancer cell line Increasing the activity of caspase, an important protein of apoptosis mechanism, was found to induce apoptosis.

In addition, as shown in Table 3, it was confirmed that it has a selective mechanism of action by showing a high affinity for the BIR3 domain of IAP, apoptosis inhibitory protein. In other words, Balb / C, a mouse normal cell line shown in Table 4, and Hs27, a human normal cell line, were not inhibited very effectively at high drug concentrations, and showed relatively high selectivity for cell lines overexpressing IAP. It can be seen that it has high selectivity for normal cell lines, the most characteristic of cell death derivatives.

Therefore, the compound of the present invention selectively affects apoptosis inhibiting protein, IAP, and has no effect on normal cells, and enables the normal mechanism of cell death of tumors and abnormal cells, thereby leading to cancer, inflammation, autoimmune diseases and neurodegenerative diseases. It can be seen that it can be used alone or in combination therapy for disorders, and has the advantage of minimizing side effects.

<110> Hanmi Co., Ltd. <120> Imidazopyrazinone derivatives with apoptosis inducing activity on          cells <130> FPD201001-0016 <160> 3 <170> KopatentIn 1.71 <210> 1 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> forward primer for XIAP BIR3 <400> 1 cgcggatcct ctgatgctgt gagttctga 29 <210> 2 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> reverse primer for XIAP BIR3 <400> 2 gagcctcgag ctaagtagtt cttaccagac 30 <210> 3 <211> 116 <212> PRT <213> Artificial Sequence <220> 223 XIAP BIR3 domain <400> 3 Ser Asp Ala Val Ser Ser Asp Arg Asn Phe Pro Asn Ser Thr Asn Leu   1 5 10 15 Pro Arg Asn Pro Ser Met Ala Asp Tyr Glu Ala Arg Ile Phe Thr Phe              20 25 30 Gly Thr Trp Ile Tyr Ser Val Asn Lys Glu Gln Leu Ala Arg Ala Gly          35 40 45 Phe Tyr Ala Leu Gly Glu Gly Asp Lys Val Lys Cys Phe His Cys Gly      50 55 60 Gly Gly Leu Thr Asp Trp Lys Pro Ser Glu Asp Pro Trp Glu Gln His  65 70 75 80 Ala Lys Trp Tyr Pro Gly Cys Lys Tyr Leu Leu Glu Gln Lys Gly Gln                  85 90 95 Glu Tyr Ile Asn Asn Ile His Leu Thr His Ser Leu Glu Glu Cys Leu             100 105 110 Val Arg Thr Thr         115

Claims (13)

A compound selected from imidazopyrazinone derivatives having the structure of Formula 1, salts, isomers, hydrates and solvates thereof:
[Formula 1]

In Chemical Formula 1,
X is oxygen or hydrogen;
R ₁ to R ₈ are each independently hydrogen, —COOR ₉ , C _1-8 alkyl, C _3-8 cycloalkyl, C _2-7 heterocycloalkyl, C _3-14 aryl, C _2-13 heteroaryl, or C _1-3 alkyl substituted with one or more C _3-14 aryl or C _2-13 heteroaryl;
The alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are each independently hydrogen, R ₉ , -OH, -OR ₉ , -COR ₉ , -COOH, -COOR ₉ , -CONR ₉ R ₁₀ , -NR ₉ , -NR ₉ CONR ₁₀ R ₁₁ , -CONR ₉ R ₁₀ , -NR ₉ CONR ₁₀ R ₁₁ , -SO ₂ R ₉ , -SO ₂ H and -SOR ₉ ;
R ₉ to R ₁₁ are each independently hydrogen, halogen, amino, nitro, carboxy, cyano, sulfyl, hydroxy, C _1-4 alkyl, C _1-4 alkoxy, C _3-8 cycloalkyl, C _{2 -7} heterocycloalkyl, C _3-14 aryl, or C _2-13 heteroaryl.
The method of claim 1,
X is oxygen or hydrogen;
R ₂ and R ₃ are each independently hydrogen, C _3-14 aryl, C _2-13 heteroaryl, or C _1-3 alkyl substituted with one or more C _3-14 aryl or C _2-13 heteroaryl;
R ₁ , R ₄ , R ₅ and R ₆ are each independently hydrogen, C _1-8 alkyl, C _3-8 cycloalkyl, or C _2-7 heterocycloalkyl;
R ₇ and R ₈ are each independently hydrogen, —COOR ₉ , C _1-8 alkyl, C _3-8 cycloalkyl, or C _2-7 heterocycloalkyl;
The alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are each independently hydrogen, R ₉ , -OH, -OR ₉ , -COR ₉ , -COOH, -COOR ₉ , -CONR ₉ R ₁₀ , -NR ₉ , -NR ₉ CONR ₁₀ R ₁₁ , -CONR ₉ R ₁₀ , -NR ₉ CONR ₁₀ R ₁₁ , -SO ₂ R ₉ , -SO ₂ H and SOR ₉ ;
R ₉ to R ₁₁ are each independently hydrogen, halogen, amino, nitro, carboxy, cyano, sulfyl, hydroxy, C _1-4 alkyl, C _1-4 alkoxy, C _3-8 cycloalkyl, C _{2 -7} heterocycloalkyl, C _3-14 aryl, or C _2-13 heteroaryl.
The method of claim 1,
X is oxygen, compound.
The method of claim 1,
R ₂ is C _1-3 alkyl substituted with one or more C _3-14 aryl or C _2-13 heteroaryl,
Wherein the aryl and heteroaryl are each independently substituted with one or more selected from the group consisting of hydrogen, halogen, C _1-4 alkyl, C _1-4 alkoxy, -COO-C _1-4 alkyl, .
The method of claim 1,
R ₃ is C _1-3 alkyl substituted with C _3-14 aryl or C _2-13 heteroaryl,
Wherein said aryl and heteroaryl are each independently substituted with one or more selected from the group consisting of hydrogen, hydroxy, C _1-4 alkyl and C _1-4 alkoxy and C _3-14 aryl.
The method of claim 5,
R ₃ is methyl substituted with phenyl, naphthyl or indolyl,
Wherein said phenyl, naphthyl and indolyl are each independently substituted with one or more selected from the group consisting of hydrogen, hydroxy, methyl, methoxy and phenyl.
The method of claim 1,
Wherein R ₁ , R ₄ , R ₅ and R ₆ are each independently hydrogen, C _1-8 alkyl, or C _3-8 cycloalkyl.
The method of claim 1,
R ₇ and R ₈ are each independently hydrogen, -COO-C _1-4 alkyl, or C _1-8 alkyl.
The method of claim 1,
Wherein said salt is a hydrochloride salt.
The method of claim 1,
A compound, characterized in that it is selected from imidazobenzopyrazinone derivatives of any one of the following compounds 1 to 47, salts, isomers, hydrates and solvates thereof:
1) (S) -N-((S) -2-((5S, 8aR) -5-benzyl-3,6-dioxo-7-phenethylhexahydroimidazo [1,2-a] pyrazine- 1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
2) (S) -N-((S) -1-cyclohexyl-2-((5S, 8aR) -5- (4-hydroxybenzyl) -3,6-dioxo-7-phenethylhexahydro Imidazo [1,2-a] pyrazin-1 (5H) -yl) -2-oxoethyl) -2- (methylamino) propanamide;
3) (S) -N-((S) -2-((5S, 8aR) -5-benzyl-7- (2,2-diphenylethyl) -3,6-dioxohexahydroimidazo [1] , 2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
4) (S) -N-((S) -2-((5R, 8aS) -5-benzyl-3,6-dioxo-7-phenethylhexahydroimidazo [1,2-a] pyrazine- 1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
5) (S) -2-amino-N-((S) -2-((5S, 8aR) -5-benzyl-3,6-dioxo-7-phenethylhexahydroimidazo [1,2- a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) propanamide;
6) (S) -N-((S) -1-cyclohexyl-2-((5S, 8aR) -5- (4-methoxybenzyl) -3,6-dioxo-7-phenethylhexahydro Imidazo [1,2-a] pyrazin-1 (5H) -yl) -2-oxoethyl) -2- (methylamino) propanamide;
7) (S) -N-((S) -1-cyclohexyl-2-((5S, 8aR) -5- (naphthalen-1-ylmethyl) -3,6-dioxo-7-phenethylhexa Hydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -2-oxoethyl) -2- (methylamino) propanamide;
8) (S) -N-((S) -1-cyclohexyl-2-((5S, 8aR) -5- (naphthalen-2-ylmethyl) -3,6-dioxo-7-phenethylhexa Hydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -2-oxoethyl) -2- (methylamino) propanamide;
9) (S) -N-((S) -2-((5S, 8aR) -5- (biphenyl-4-ylmethyl) -3,6-dioxo-7-phenethylhexahydroimidazo [ 1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
10) ((S) -N-((S) -1-cyclohexyl-2-((5S, 8aR) -5- (4-methylbenzyl) -3,6-dioxo-7-phenethylhexahydro Imidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
11) (S) -N-((S) -2-((5S, 8aR) -5-benzyl-3-oxo-7-phenethylhexahydroimidazo [1,2-a] pyrazine-1 (5H ) -Yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
12) (S) -N-((S) -2-((5S, 8aR) -5-benzyl-3,6-dioxo-7- (3-phenylpropyl) hexahydroimidazo [1,2- a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
13) (S) -N-((S) -2-((5S, 8aR) -5-benzyl-7- (4-fluorophenethyl) -3,6-dioxohexahydroimidazo [1, 2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
14) (S) -N-((S) -3,3-dimethyl-1-((5S, 8aR) -5- (naphthalen-1-ylmethyl) -3,6-dioxo-7-phenethyl Hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-oxobutan-2-yl) -2- (methylamino) propanamide;
15) (S) -N-((S) -1-cyclohexyl-2-((2S, 5S, 8aR) -2-methyl-5- (naphthalen-1-ylmethyl) -3,6-dioxo -7-phenethylhexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -2-oxoethyl) -2- (methylamino) propanamide;
16) (S) -N-((S) -2-((5S, 8aR) -7-benzyl-5- (naphthalen-2-ylmethyl) -3,6-dioxohexahydroimidazo [1, 2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
17) (S) -N-((S) -2-((5S, 8aR) -7- (4-chlorobenzyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxohexahydro Imidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
18) (S) -N-((S) -2-((5S, 8aR) -7- (4-methoxybenzyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxohexahexa Hydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
19) (S) -N-((S) -2-((5S, 8aR) -7- (4-methoxyphenethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxo Hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
20) (S) -N-((S) -2-((5S, 8aR) -7- (4-fluorophenethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxo Hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
21) (S) -N-((S) -2-((5S, 8aR) -7- (4-fluorophenethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxo Hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;
22) (S) -N-((S) -2-((5S, 8aR) -7- (4-fluorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahydro Imidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
23) (S) -N-((S) -2-((5S, 8aR) -7- (4-fluorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahydro Imidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;
24) (S) -N-((S) -2-((5S, 8aR) -7- (3-chlorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahydroimide Dazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
25) (S) -N-((S) -2-((5S, 8aR) -7- (2-chlorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahydroimide Dazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
26) (S) -N-((S) -2-((5S, 8aR) -5,7-bis (4-methylbenzyl) -3,6-dioxohexahydroimidazo [1,2-a ] Pyrazin-1 (5H) -yl-3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;
27) (S) -N-((S) -1-((5S, 8aR) -7- (3-chlorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahydroimide Dazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;
28) (S) -N-((S) -2-((5S, 8aR) -7- (4-pyridinemethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxohexahydro Imidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
29) (S) -N-((S) -2-((5S, 8aR) -7- (4-pyridinemethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxohexahydro Imidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;
30) (S) -N-((S) -2-((5S, 8aR) -7- (methyl- (4-methylbenzoate))-5- (naphthalen-2-ylmethyl) -3,6 -Dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
31) (S) -N-((S) -2-((5S, 8aR) -7- (methyl- (4-methylbenzoate))-5- (naphthalen-2-ylmethyl) -3,6 -Dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;
32) (S) -N-((S) -1-((5S, 8aR) -7- (2-chlorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahydroimide Dazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;
33) (S) -N-((S) -3,3-dimethyl-1-((5S, 8aR) -5- (naphthalen-2-ylmethyl) -3,6-dioxo-7-phenethyl Hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -1-oxobutan-2-yl) -2- (methylamino) propanamide;
34) (S) -N-((S) -2-((5S, 8aR) -7- (4-chlorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahydroimide Dazo [1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
35) (S) -N-((S) -1-((5S, 8aR) -7- (4-chlorophenethyl) -5- (4-methylbenzyl) -3,6-dioxohexahydroimide Dazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;
36) Methyl 4-(((5S) -5- (biphenyl-4-ylmethyl) -1-((S) -2-cyclohexyl-2-((S) -2- (methylamino) propaneami Acetyl) -3,6-dioxohexahydroimidazo [1,2-a] pyrazin-7 (1H) -yl) methyl) benzoate;
37) (S) -N-((S) -2-((5S, 8aR) -7- (3-chlorophenethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxohexa Hydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;
38) (S) -N-((S) -2-((5S, 8aR) -7- (2-chlorophenethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxohexa Hydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;
39) (S) -N-((S) -2-((5S, 8aR) -7- (2-phenylpropyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxohexahydro Imidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;
40) (2S) -N-((2S) -1-((5S) -5-((1H-indol-3-yl) methyl) -3,6-dioxo-7-pentylhexahydroimidazo [ 1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl-2- (methylamino) propanamide;
41) (2S) -N-((1S) -2-((5S) -5-((1H-indol-3-yl) methyl) -3,6-dioxo-7-pentylhexahydroimidazo [ 1,2-a] pyrazin-1 (5H) -yl) -1-cyclohexyl-2-oxoethyl) -2- (methylamino) propanamide;
42) ((S) -N-((S) -2-((5S, 8aR) -7- (2-fluorophenethyl) -5- (naphthalen-2-ylmethyl) -3,6-di Oxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;
43) (S) -N-((S) -2-((5S, 8aR) -7- (2-bromophenethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxohexa Hydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;
44) (S) -N-((S) -2-((5S, 8aR) -7- (2-methoxyphenethyl) -5- (naphthalen-2-ylmethyl) -3,6-dioxo Hexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino) propanamide;
45) (S) -N-((S) -2-((5S, 8aR) -7- (2,3-dihydro-1H-inden-2-yl) -5- (naphthalen-2-ylmethyl ) -3,6-dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methylamino Propanamide;
46) (S) -N-((S) -2-((5S, 8aR) -7- (4-methyl-1,2,3,4-tetrahydronaphthalen-2-yl) -5- (naphthalene -2-ylmethyl) -3,6-dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl)- 2- (methylamino) propanamide; And
47) (S) -N-((S) -2-((5S, 8aR) -7- (1,2,3,4-tetrahydronaphthalen-2-yl) -5- (naphthalen-2-yl Methyl) -3,6-dioxohexahydroimidazo [1,2-a] pyrazin-1 (5H) -yl) -3,3-dimethyl-1-oxobutan-2-yl) -2- (methyl Amino) propanamide.
A pharmaceutical composition comprising at least one of the compounds according to claim 1, salts, isomers, hydrates and solvates thereof as an active ingredient, together with a pharmaceutically acceptable carrier.
The method of claim 11,
The pharmaceutical composition is characterized in that for preventing and treating cancer, inflammation, autoimmune diseases or neurodegenerative disorders.
A compound library comprising at least one of the compound according to claim 1, salts, isomers, hydrates and solvates thereof.