WO2007142431A1

WO2007142431A1 - New non-peptide compounds, process for the preparation thereof and pharmaceutical composition comprising the same

Info

Publication number: WO2007142431A1
Application number: PCT/KR2007/002663
Authority: WO
Inventors: Hea-Young Park Choo; Heung Sik Na; Yoo Lim Kam; Hee Kyung Rhee; Seung Keun Back
Original assignee: Ewha University - Industry Collaboration Foundation
Priority date: 2006-06-02
Filing date: 2007-06-01
Publication date: 2007-12-13
Also published as: KR20070115768A; KR100888069B1

Abstract

The present invention relates to a novel non-peptide compound, a method for preparing the same, and a pharmaceutical composition comprising the same. The non-peptide compound of the present invention has excellent inhibitory effect on bradykinin activity, and on chronic neuropathic pain and inflammatory pain, thereby being used for preventing or treating a disease caused by bradykinin, in particular, pain.

Description

NEW NON-PEPTIDE COMPOUNDS, PROCESS FOR THE

PREPARATION THEREOF AND PHARMACEUTICAL

COMPOSITION COMPRISING THE SAME

Technical Field

[1] The present invention relates to a novel non-pep tide compound, a method for preparing the same, and a pharmaceutical composition comprising the same. Background Art

[2] Bradykinin is an endogenous peptide hormone consisting of 9 amino acids, and produced from its kininogen precursor by kallikrein, which belongs to a group of proteolytic enzymes. Bradykinin is released when inflammation, trauma, burn injury, shock, allergy, and specific cardiovascular disease occur. Once released, bradykinin initiates or increases the secretion of a mediator, which stimulates sensory afferent nerve endings, from leukocyte. Bradykinin causes a variety of pathophysiological responses such as septic shock, hemorrhagic shock, hypersensitivity, arthritis, rhinitis, asthma, inflammatory bowel disease, acute pancreatitis, post-gastrectomy dumping syndrome, carcinoid syndrome, migraine, and hereditary angioneurotic edema.

[3] Bradykinin receptors are mainly classified into B 1 type and B2 type according to efficacy of various bradykinin analogs. Bradykinin acts on the Bl receptor to contract veins and arteries mainly, although it can cause relaxation of peripheral resistance vessels as well. However, more important functions such as increased vascular permeability, pain, and vasodilatation, are mediated by the B2 receptor. Bradykinin acts on the B2 receptor to cause hypotension and inflammation, and it increases vascular permeability to cause edema and pain. Thus, these effects of bradykinin on the B2 receptor are believed to be responsible for numerous diseases, such as inflammation, cardiovascular disease, pain, and the common cold. Accordingly, B 1 receptor antagonists can be used as a drug having anti-inflammatory and anti-pyretic effects, and B2 receptor antagonists can be used as an agent for treating diseases such as asthma, allergic rhinitis, the common cold, inflammatory joint disease, pancreatitis, and cystitis.

[4] The first generation of bradykinin antagonists is a peptide antagonist, in which the proline at position 7 of an amino acid sequence was substituted with D-phenylalanine or D-aromatic amino acid. Its representative material is D-Arg-[Hyp 3, Thi 5,8, D-Phe 7]BK, which has been known as NPC-349 [Vaverk, R. J.; Sewart, J. M. Competitive antagonists of bradykinin Peptides 1985, 6, 161-164]. The second generation of bradykinin antagonist developed is Icatibant (HOE 140) and Brady cor (CPO 127, dimeric peptide) [Griesbacher, T.; Lembeck, F.; Eckhardt, M.; Henke, St.; Breipohl, G.; Knolle, J. New, long-acting, potent bradykinin antagonists. Br. J. Pharmacol. 1991, 102, 257-304.; Hock, F. J.; Wirth, K.; Albus, U.; Linz, W.; Gerhard, H. J.; Weimer, G.; Henke, S.; Breiphl, G.; Konig, W.; Knolle, J.; Scholkens, B. A. HOE 140, a new potent and long acting bradykinin-antagonist: In vitro studies. Br. J. Pharmacol. 1991, 102, 769-773.; Wirth, K.; Albus, U.; Linz, W.; Alpermann, H. J.; Anagnostopoulos, H.; Henke, S.; Breiphl, G.; Konig, W.; Knolle, J.; Scholkens, B. A. HOE 140, a new potent and long acting bradykinin-antagonist: In vivo studies. Br. J. Pharmacol. 1991, 102, 77 4-777]. The second generation of bradykinin antagonists have stronger affinity for the B2 receptor, as compared to the first generation of bradykinin antagonists, and have longer duration of action in vivo. However, they have poor bioavailability when administered orally, thus their clinical application is still limited.

[5] Recently, a lot of studies have been conducted on a non-peptide antagonist of a bradykinin receptor, which can be orally administered for a long period of time to treat chronic inflammatory diseases. Representative examples thereof are WIN 64338 and FR 173657, which are phosphonium derivative and heteroaryl benzyl ether, respectively [Salvino, J. M.; Seoane, P. R.; Douty, B. D.; Awad, M. M. A.; Dolle, R. E.; Houck, W. T.; Faunce, D. M.; Sawutz, D. G. Design of potent non-peptide competitive antagonist of the human bradykinin B2 receptor. J. Med. Chem. 1993, 36, 2583-2584, Abe, Y.; Kayakiri, H.: Satoh, S.; Inoue, T.; Sawada, Y.; Inamura, N.; Asanno, M.; Hatori, C; Oku, T.; Tanaka, H. A novel class of orally active non-peptide bradykinin B2 receptor antagonists. Discovering bioisosteres of the Imidazo[l,2-a]pyridine moiety. J. Med. Chem. 1998, 41, 4062-4079]. Further, the antagonistic effect on the B2 receptor was slightly observed in some compounds among the second generation of antihistamine having a piperazine residue. In particular, antihistamine and antibradykinin effects were observed by modifying a carbon side chain in cetirizine into an aryl or alkyl chain. In accordance with this, the present inventors developed a compound having excellent antibradykinin effect among the compounds having a piperazine residue (Korean Patent No. 10-0446711). However, there is a need in the art to develop a compound having a much better antibradykinin effect, in particular, a compound having excellent analgesic effect, as well as antibradykinin effect. Disclosure of Invention Technical Problem

[6] Therefore, the present inventors have made extensive studies on the compound having excellent analgesic effect as well as a much better antibradykinin effect, so as to prepare a novel non-peptide compound. They have found that the compound effectively inhibits bradykinin activity and has an excellent inhibitory effect on chronic neuropathic pain and inflammatory pain, thereby completing the present invention. Technical Solution

[7] The present invention relates to a novel non-pep tide compound, a method for preparing the same, and a pharmaceutical composition comprising the same, for preventing or treating a disease caused by bradykinin.

Brief Description of the Drawings [8] Fig. 1 is a drawing showing a step of transecting the inferior/superior caudal trunks between the S 1 and S2 spinal nerves that innervate a rat tail in order to induce chronic neuropathic pain. [9] Fig. 2 is a drawing showing the inhibitory effect of the compound (Example 6) of the invention on chronic neuropathic pain (A: mechanical allodynia, B: cold allodynia, and C: warm allodynia). [10] Fig. 3 is a drawing showing the inhibitory effect of the compound (Example 26) of the invention on chronic neuropathic pain (A: mechanical allodynia, B: cold allodynia, and C: warm allodynia). [11] Fig. 4 is a drawing showing the inhibitory effect of the compound (Example 3) on inflammatory pain. [12] Fig. 5 is a drawing showing the inhibitory effect of the compound (Example 6) on inflammatory pain. [13] Fig. 6 is a drawing showing the inhibitory effect of the compound (Example 14) on inflammatory pain. [14] Fig. 7 is a drawing showing the inhibitory effect of the compound (Example 20) on inflammatory pain. [15] Fig. 8 is a drawing showing the inhibitory effect of the compound (Example 26) on inflammatory pain. [16] Fig. 9 is a drawing showing the inhibitory effect of the compound (Example 27) on inflammatory pain.

Best Mode for Carrying Out the Invention [17] The present invention provides a non-peptide compound represented by the following formula 1. [18] [Formula 1]

[19]

[20] wherein

[21] Rl and R2 are each independently hydrogen; C ~C alkyl; C ~C alkoxy; C ~C

1 4 1 4 3 10 cycloalkyl; C ~C aryl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; C ~C heteroaryl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; C ~C alkyl substituted with C ~C aryl which is substituted or unsubstituted ^to 1 4 ^J 6 20 ^J with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; or C ~C alkyl substituted with C ~C heteroaryl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen, [22] X is C ~C alkylenyl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; C ~C alkenylenyl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; -CH 2 YCH 2 -; or -CH 2 CH 2 Y-,

[23] Y is O, S, or NR3,

[24] R3 is hydrogen, C ~C alkyl, C ~C alkoxy, halogen or -COO-C ~C alkyl,

[25] Z and Z' are each independently hydrogen, hydroxy, or halogen, and

[26] n is an integer of 1 to 3

[27] (provided that if n is 2, X is not -CH YCH - (wherein Y is NR3, and R3 is hydrogen)). [28]

[29] Preferably, in the formula 1,

[30] Rl and R2 are each independently hydrogen; C ~C alkyl; C ~C cycloalkyl; or C

~C alkyl substituted with C ~C aryl which is substituted or unsubstituted with at

4 ^J 6 20 ^J least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen,

[31] X is C ~C alkylenyl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; C ~C alkenylenyl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; or -CH YCH -,

[32] Y is NR3,

[33] R3 is hydrogen or -COO-C ~C alkyl,

[34] Z and Z' are each independently hydrogen or halogen, and

[35] n is an integer of 2 or 3

[36] (provided that if n is 2, X is not -CH YCH - (wherein Y is NR3, and R3 is hydrogen)). [37]

[38] More preferably, in the formula 1,

[39] Rl and R2 are each independently hydrogen, methyl, cyclohexyl, 4-methylbenzyl,

4-methoxybenzyl, 3,4,5-trimethoxybenzyl, or l-methyl-3-phenylpropyl, [40] X is ethylenyl, 2-methylpropylenyl, 2,2-dimethylpropylenyl, ethenylenyl, -CH

NHCH -, or -CH NCOO(t-butyl)CH -,

2 2 2

[41] Z and Z' are each independently hydrogen, F, or Cl, and

[42] n is an integer of 2 or 3

[43] (provided that if n is 2, X is not -CH NHCH -).

[44]

[45] Specific examples of the non-pep tide compounds represented by the formula 1 are preferably as follows: [46] 1)

5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-N-(4-methylbenzyl)-3-methyl-5

-oxopentanamide; [47] 2)

5 - (4- ( (4-chlorophenyl) (phenyl)methyl)piperazin- 1 -yl) -N- (4-methoxybenzyl) - 3 -methyl-

5 -oxopentanamide ; [48] 3)

5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-N-(3,4,5-trimethoxybenzyl)-3-m ethyl- 5 -oxopentanamide ; [49] 4)

5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-N-cyclohexyl-3-methyl-5-oxope ntanamide; [50] 5)

5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-N,N,3-trimethyl-5-oxopentanam ide; [51] 6)

5-(4-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan-l-yl)-N-(4-methylbenzyl)-3-meth yl- 5 -oxopentanamide ; [52] 7)

5-(4-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan-l-yl)-N-(4-methoxybenzyl)-3-me thyl- 5 -oxopentanamide ; [53] 8)

5-(4-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan-l-yl)-N-(3,4,5-trimethoxybenzyl)

-3-methyl-5-oxopentanamide; [54] 9) 5-(4-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan-l-yl)-N-cyclohexyl-3-methyl-5-o xopentanamide; [55] 10) t-butyl (dimethylcarbamoyl)methyl

2-(4-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan-l-yl)-2-oxoethylcarbamate; [56] 11)

2-(2-(4-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan-l-yl)-2-oxoethylamino)-N,N-d imethylacetamide ;

[57] 12) N-(4-methylbenzyl)-4-oxo-4-((4-chlorobenzhydryl)piperazin- l-yl)butanamide;

[58] 13) N-

(4-methoxybenzyl)-4-oxo-4-((4-chlorobenzhydryl)piperazin-l-yl)butanamide; [59] 14) N-(3,4,5-trimethoxybenzyl)-4-oxo-4-((4-chlorobenzhydryl)piperazin-l-yl) butanamide;

[60] 15) N-(cyclohexyl)-4-oxo-4-((4-chlorobenzhydryl)piperazin-l-yl)butanamide;

[61] 16) N-

(4-methylbenzyl)-4-oxo-4-((4-chlorobenzhydryl)- 1 ,4-diazepan- 1 -yl)butanamide; [62] 17) N-

(4-methoxybenzyl)-4-oxo-4-((4-chlorobenzhydryl)-l,4-diazepan-l-yl)butanamide; [63] 18) N-

(3,4,5-trimethoxybenzyl)-4-oxo-4-((4-chlorobenzhydryl)-l,4-diazepan-l-yl)butanamid e;

[64] 19) N-(cyclohexyl)-4-oxo-4-((4-chlorobenzhydryl)-l,4-diazepan-l-yl)butanamide;

[65] 20) N-

(3,4,5-trimethoxybenzyl)-N'- 1 -((4-chlorobenzhydryl)- 1 ,4-diazepan- 1 -yl)iminodiacetic acid diamide; [66] 21)

(Z)-N-(4-methylbenzyl)-4-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-4-oxo- but-2-enamide; [67] 22)

(Z)-N-(4-methoxybenzyl)-4-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-4-oxo

-but-2-enamide; [68] 23)

(Z)-4-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-4-oxo-N-(4-phenylbutan-2- yl)-but-2-enamide; [69] 24)

(Z)-N-(4-methylbenzyl)-4-(4-(bis(4-fluorophenyl)methyl)piperazin-l-yl)-4-oxo-but-2- enamide; [70] 25)

(Z)-N-(4-methoxybenzyl)-4-(4-(bis(4-fluorophenyl)methyl)piperazin-l-yl)-4-oxo-but- 2-enamide;

[71] 26)

(Z)-4-(4-(bis(4-fluorophenyl)methyl)piperazin-l-yl)-4-oxo-N-(4-phenylbutan-2-yl)-but -2-enamide;

[72] 27)

2-(2-(4-(bis(4-fluorophenyl)methyl)piperazin-l-yl)-2-oxoethylamino)-N-(4-phenylbuta n-2-yl)acetamide;

[73] 28)

2-(2-(4-(bis(4-fluorophenyl)methyl)piperazin-l-yl)-2-oxoethylamino)-N-(3,4,5-trimeth oxybenzyl)acetamide;

[74] 29) N-

(4-methylbenzyl)-5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-3,3-dimethyl- 5 -oxopentanamide ;

[75] 30) N-

(4-methoxybenzyl)-5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-3,3-dimethy 1-5 -oxopentanamide ;

[76] 31) N-

(3,4,5-trimethoxybenzyl)-5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-3,3-di methyl- 5 -oxopentanamide; and

[77] 32) N- cyclohexyl-5 - (4- ( (4-chlorophenyl) (phenyl)methyl)piperazin- 1 -yl) -3 , 3 -dimethyl- 5 -oxop entanamide.

[78]

[79] The non-peptide compound represented by the formula 1 of the invention may be used in the form of a pharmaceutically acceptable salt, and as the salt, acid addition salts prepared with pharmaceutically acceptable free acids are preferred. As the free acids, inorganic acids and organic acids may be used. Examples of the inorganic acids include hydrochloric acid, bromic acid, sulfuric acid, and phosphoric acid, and examples of the organic acids include citric acid, acetic acid, lactic acid, tartaric acid, fumaric acid, formic acid, propionic acid, oxalic acid, trifluoroacetic acid, methanesulfonic acid, benzenesulfonic acid, maleic acid, benzoic acid, gluconic acid, glycolic acid, succinic acid, 4-morpholineethanesulfonic acid, camphorsulfonic acid, 4-nitrobenzenesulfonic acid, hydroxy-O-sulfonic acid, 4-toluenesulfonic acid, galacturonic acid, embonic acid, glutamic acid, and aspartic acid.

[80]

[81] Further, the present invention provides a method comprising the steps of

[82] 1) preparing a compound represented by the following formula 4 by condensation reaction of a compound represented by the following formula 2 and an amine compound represented by the following formula 3; and [83] 2) preparing the compound represented by the formula 1 by condensation reaction of the compound represented by the following formula 4 and a compound represented by the following formula 5. [84] [Formula 2]

[85]

[86] [Formula 3]

[87] R1R2NH

[88] [Formula 4] [89]

O O

^R1-NΛAOH

R2

[90] [Formula 5]

[91]

[92] wherein

[93] Rl and R2 are each independently hydrogen; C ~C alkyl; C ~C alkoxy; C ~C

1 4 1 4 3 10 cycloalkyl; C ~C aryl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; C ~C heteroaryl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halog ^toen; C 1 ~C 4 alky ^Jl substituted with C 6 ~C 20 ary ^Jl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; or C ~C alkyl substituted with C ~C heteroaryl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C 1 ~C4 alkyl, C 1 ~C4 alkoxy, and halogen,

[94] X is C ~C alkylenyl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; C ~C alkenylenyl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; -CH YCH -; or -CH CH Y-,

^& 2 2 2 2

[95] Y is O, S, or NR3,

[96] R3 is hydrogen, C ~C alkyl, C ~C alkoxy, halogen, or -COO-C ~C alkyl,

[97] Z and Z' are each independently hydrogen, hydroxy or halogen, and

[98] n is an integer of 1 to 3

[99] (provided that if n is 2, X is not -CH YCH - (wherein Y is NR3, and R3 is hydrogen)).

[100]

[101] The compound represented by the formula 2 used in the above-described preparation method is preferably 3-methylglutaric anhydride, or succinic anhydride. Further, in the compound represented by the formula 2, if X is -CH YCH - or -CH CH Y-, and Y is NR3, R3 represents an amino-protecting group, preferably t- butyloxycarbonyl.

[102] The amine compound represented by the formula 3 is preferably

4-methylbenzylamine, 4-methoxybenzylamine, 3,4,5-trimethoxybenzylamine, cyclo- hexylamine, dimethylamine, or l-methyl-3-phenylpropylamine.

[103] The compound represented by the formula 5 is preferably

1 - (4-chlorobenzhydryl)piperazine, or l-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan.

[104]

[105] In the steps 1) and 2), the condensation reaction can be performed under the reaction condition for peptide bond known in the art. Examples of the reaction solvent used in the reaction include ethyl ether, THF (tetrahydrofuran), dichloromethane, chloroform, DMSO (dimethyl sulfonyloxide), and DMF (dimethylformamide), preferably DMF.

[106] In the step 2), in order to promote the reaction, the peptide bond reaction can be performed with a coupling agent, which is known in the art, as a catalyst, in the presence or nonpresence of a base such as TEA (triethylamine), DIEA (N,N-diisopropylethylamine), and NMM (N-methyl morpholine). Examples of the coupling agent include DIC (diisopropyl carbodiimide), EDC( 1 -(3-dimethylaminopropyl)-3-ethylcarbodiimide),

TBTU(2-( 1 -H-benzotriazol- 1 -yl)- 1 , 1 ,3,3-tetramethyluromium tetrafluoro borate), DCC(dicyclohexyl carbodiimide), HATU (dimethylamino-([l,2,3]triazolo[4,5-b] pyridin-3-yloxy)-methylene)-dimethyl-ammonium hexafluorophosphate), and PyBOP(benzotriazol-l-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate). Preferably, PyBOP can be used as a reaction catalyst in the presence of DIEA.

[107] In the compound of the formula 2, if X is nitrogen containing a protecting group, the compound is reacted with HCl-dioxane and chloroform to perform a deprotecting reaction after the step 2).

[108]

[109] Further, the present invention provides a pharmaceutical composition comprising the non-peptide compound of the formula 1, or a pharmaceutically acceptable salt thereof, for preventing or treating a disease caused by bradykinin.

[110] The compound according to the invention has excellent inhibitory effect on bradykinin activity, and on chronic neuropathic pain and inflammatory pain, thereby being used for preventing or treating a disease caused by bradykinin, in particular, pain.

[I l l] Examples of the disease caused by bradykinin include asthma, allergic rhinitis, arthritis, rheumatic arthritis, septic shock, hemorrhagic shock, hypersensitivity, cerebral edema, angioneurotic edema, acute pancreatitis, post-gastrectomy dumping syndrome, carcinoid syndrome, migraine, and pain.

[112] The composition of the invention may contain at least one kind of active ingredient known in the art, which has the effect of preventing or treating the disease caused by bradykinin, in addition to the non-peptide compound of the formula 1.

[113] For administration, the composition of the invention can be prepared including at least one pharmaceutically acceptable carrier, in addition to the active ingredients as described above. Examples of the pharmaceutically acceptable carrier include a saline solution, sterile water, a Ringer's solution, a buffered saline solution, a dextrose solution, a maltodextrin solution, glycerol, ethanol and a mixture of two or more thereof. If necessary, the composition may also contain other conventional additives, such as antioxidants, buffers, and bacteriostatic agents. Moreover, the composition may additionally contain diluents, dispersants, surfactants, binders, and lubricants in order to formulate it into injectable formulations, such as aqueous solutions, suspensions, and emulsions, pills, capsules, granules and tablets. Furthermore, the composition may preferably be formulated depending on the types of diseases and its components, using the method described in Remington's Pharmaceutical Science (latest edition), Mack Publishing Company, Easton Pa., which is a suitable method in the relevant field of art.

[114] The composition of the invention may be administered orally or parenterally (for example, intravenous, subcutaneous, intraperitoneal, or topical application) depending on the purpose of the invention, and the dosage of the composition can vary depending on various factors, including patient's weight, age, sex, health condition, and diet, and administration time, administration route, secretion rate, disease severity, etc. The compound of the formula 1 is administered at a daily dosage of about 5 to 250 mg/kg, preferably 8 to 60 mg/kg once or several times. The composition of the invention may be used alone or in combination with surgical operations, hormone therapies, chemical therapies, and other methods using biological reaction regulators for preventing or treating a disease caused by bradykinin. Mode for the Invention

[115] Hereinafter, preferred Examples are provided for the purpose of illustrating the present invention. However, these Examples are provided for the illustrative purpose only, and the invention is not intended to be limited by these Examples.

[116] Preparative Example 1 : Preparation of

4-(4-methylbenzylcarbamoyl)-3-methylbutanoic acid

[117] 2 mmol of 3-methylglutaric anhydride was added to 3 mL of dimethylformamide

(DMF) to dissolve. Then, 2 mmol of 4-methylbenzylamine was added thereto, and stirred at room temperature for 20 hours. 20 mL of 10% HCl was put into the reaction solution, and extracted with 30 mL of ethylacetate (EtOAc). The organic layer was washed with 20 mL of 10% HCl, and then washed with 20 mL of a saturated NaCl solution twice. The organic layer was collected, dried over anhydrous MgSO , and

4 filtered under reduced pressure. The organic solvent in the filtrate was removed under reduced pressure. The residue was recrystallized from a mixed solvent of EtOAc and methanol, so as to obtain the title compound (white solid, yield: 96%). [118] mp l33-134°C;

[119] ¹U NMR (CDCl ) δ 7.16 (s, 4CH), 5.90 (s, NH), 4.41 (d, J=6.0 Hz, CH ), 2.43-2.46

(m, CH), 2.40 (t, J=7.2 Hz, CTT), 2.34 (s, CH₃), 2.28 (t, J=6.4 Hz, CTT), 1.08 (d, J=6.4

Hz, CH₃);

[120] HR-FABMS Calcd for C₁₄H₁ NO₃ (M⁺+l): 250.1443, Found: 250.1444.

[121] [122] Preparative Example 2 : Preparation of

4-(4-methoxybenzylcarbamoyl)-3-methylbutanoic acid [123] The title compound was prepared in the same manner as in Preparative Example 1, except that 4-methoxybenzylamine was used instead of 4-methylbenzylamine in

Preparative Example 1 (white solid, yield: 74%). [124] mp 89-9O⁰C;

[125] ¹U NMR (CDCl ) δ 7.20 (d, J=9.2 Hz, 2CH), 6.87 (d, J=9.2 Hz, 2CH), 5.94 (s,

NH), 4.38 (d, J=5.6 Hz, CH ), 3.80 (s, OCH ), 2.36-2.48 (m, CH), 2.32 (dd, J=6.4 Hz,

CH₂), 2.23 (dd, J=6.4 Hz, CH₂), 1.06 (d, J=6.4 Hz, CH₃); [126] HR-FABMS Calcd for C H NO (M⁺+l): 266.1392, Found: 266.1382.

14 19 4

[127] [128] Preparative Example 3 : Preparation of

4-(3,4,5-trimethoxybenzylcarbamoyl)-3-methylbutanoic acid

[129] The title compound was prepared in the same manner as in Preparative Example 1, except that 3,4,5-trimethoxybenzylamine was used instead of 4-methylbenzylamine in

Preparative Example 1 (white solid, yield: 28%). [130] mp l l8-119°C;

[131] ¹U NMR (CDCl ) δ 6.50 (s, 2CH), 4.39 (d, J=5.6 Hz, CH ), 3.85 (s, 3OCH ),

2.41-2.48 (m, CH), 2.45 (d, J=4.4 Hz, CH₂), 2.31 (d, J=5.6 Hz, CH₂), 1.10 (d, J=6.8

Hz, CH ); [132] HR-FABMS Calcd for C 16 H 23 NO 6 (M⁺+l): 326.1604, Found: 326.1606.

[133]

[134] Preparative Example 4 : Preparation of

4-(cyclohexylcarbamoyl)-3-methylbutanoic acid

[135] The title compound was prepared in the same manner as in Preparative Example 1, except that cyclohexylamine was used instead of 4-methylbenzylamine in Preparative Example 1 (white solid, yield: 69%).

[136] mp 133-134⁰C;

[137] ¹H NMR (CDCy δ 5.56 (s, NH), 3.73-3.84 (m, CH), 2.35-2.45 (m, CH), 2.30; 2.27

(dd, J=5.6; 6.0 Hz, CH₂), 2.24; 2.21 (dd, J=7.2 Hz, CH ), 1.93 (d, /=12.0 Hz, CH₂), 1.72 (d, /=12.4 Hz, CH ), 1.63 (d, /=12.8 Hz, CH ), 1.41; 1.34 (dd, /=12.0 Hz, CH ), 1.16 (t, /=11.6 Hz, CH₂), 1.08 (d, /=6.4 Hz, CH₃);

[138] HR-FABMS Calcd for C 12 H 21 NO 3 (M⁺+ 1): 228.1600, Found: 228.1601.

[139]

[140] Preparative Example 5 : Preparation of

4-(dimethylcarbamoyl)-3-methylbutanoic acid

[141] The title compound was prepared in the same manner as in Preparative Example 1, except that dimethylamine was used instead of 4-methylbenzylamine in Preparative Example 1 (white solid, yield: 2%).

[142] mp l l6-117°C;

[143] ¹U NMR (CDCl ) δ 3.05 (s, CH ), 2.98 (s, CH ), 2.35-2.49 (m, CH), 2.33 2.31 (dd,

/=5.6 Hz, CH₂), 2.27; 2.23 (dd, /=7.6; 7.2 Hz, CH₂), 1.08 (d, /=6.0 Hz, CH₃);

[144] HR-FABMS Calcd for C 8 H 15 NO 3 (M⁺+Na): 196.0950, Found: 196.0949.

[145]

[146] Example 1 : Preparation of

5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-N-(4-methylbenzyl)-3-meth yl-5-oxopentanamide

[147]

[148] 0.5 mmol of 4-(4-methylbenzylcarbamoyl)-3-methylbutanoic acid prepared in

Preparative Example 1, 0.55 mmol of l-(4-chlorobenzhydryl)piperazine, and ben- zotriazol-1-yloxytripyrrolidino phosphonium hexafluorophosphate (PyBOP) were added to 3 mL of dimethylformamide (DMF) to dissolve. Then, 1.0 mmol of N,N-diisopropylethylamine (DIEA) was added thereto, and stirred at room temperature for 16 hours. 20 mL of 10% HCl was put into the reaction solution, and extracted with 30 mL of EtOAc. The organic layer was washed with 20 mL of 10% HCl, and then washed with 20 mL of a saturated NaHCO solution twice and with 20 mL of a saturated NaCl solution twice. The organic layer was collected, dried over anhydrous MgSO 4 , and filtered under reduced pressure. The organic solvent in the filtrate was removed under reduced pressure. The residue was purified by column chromatography with a mixed solvent of EtOAc and methanol (20:1), so as to obtain the title compound (light yellow solid, yield: 65%).

[149] mp 62-63⁰C;

[150] ¹U NMR (CDCl ) δ 7.35 (d, J=8.0 Hz, 2CH), 7.29 (d, J=7.2 Hz, 2CH), 7.26 (t, J

=8.2 Hz, 2CH), 7.22 (t, J=7.3 Hz, CH), 7.16 (d, J=8.0 Hz, 2CH), 7.11 (d, J=7.2 Hz, 2CH), 6.29 (s, NH), 4.37 (d, J=5.6 Hz, CH₂), 4.21 (s, CH), 3.59 (t, J=5.6 Hz, CH₂), 3.48 (q, J=5.6 Hz, CH₂), 2.43 (d, J=6.8 Hz, CH₂), 2.34-2.39 (m, CH₂), 2.32 (s, CH₃), 2.28 (d, J=6.0 Hz, CH ), 2.25 (d, J=6.4 Hz, CH ), 2.13-2.23 (m, CH), 1.03 (d, J=6.4 Hz, CH₃);

[151] HR-FABMS Calcd for C H ClN O : (M⁺+l): 518.2574, Found: 518.2584.

31 36 3 2

[152]

[153] Example 2 : Preparation of

5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-N-(4-methoxybenzyl)-3-inet hyl-5-oxopentanamide

[154]

[155] The title compound was prepared in the same manner as in Example 1, except that

4-(4-methoxybenzylcarbamoyl)-3-methylbutanoic acid prepared in Preparative Example 2 was used instead of 4-(4-methylbenzylcarbamoyl)-3-methylbutanoic acid in Example 1 (white solid, yield: 63%). [156] mp 61-62⁰C;

[157] ¹U NMR (CDCl ) δ 7.35 (d, J=8.4 Hz, 2CH), 7.29 (d, J=7.6 Hz, 2CH), 7.26 (t, J

=8.2 Hz, 2CH), 7.23 (d, J=7.6 Hz, CH), 7.20 (d, J=8.8 Hz, 2CH), 6.84 (d, J=8.0 Hz, 2CH), 6.31 (s, NH), 4.35 (d, J=5.6 Hz, CH₂), 4.21 (s, CH), 3.79 (s, OCH₃), 3.59 (t, J =5.0 Hz, CH ), 3.48 (q, J=5.2 Hz, CH ), 2.42 (d, J=7.2 Hz, CH ), 2.34-2.38 (m, CH ), 2.28 (d, J=6.4 Hz, CH₂), 2.25 (d, J=6.0 Hz, CH₂), 2.18-2.22 (m, CH), 1.03 (d, J=6.8 Hz, CH );

[158] HR-FABMS Calcd for C 31 H 36 ClN 303 : (M⁺+l): 534.2523, Found: 534.2528.

[159]

[160] Example 3 : Preparation of

5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-N-(3,4,5-trimethoxybenzyl)-

3-methyl-5-oxopentanamide

[161]

[162] The title compound was prepared in the same manner as in Example 1, except that

4-(3,4,5-trimethoxybenzylcarbamoyl)-3-methylbutanoic acid prepared in Preparative Example 3 was used instead of 4-(4-methylbenzylcarbamoyl)-3-methylbutanoic acid in Example 1 (white solid, yield: 71%).

[163] mp 72-73°C;

[164] ¹U NMR (CDCl ) δ 7.35 (d, J=8.4 Hz, 2CH), 7.29 (d, J=6.8 Hz, 2CH), 7.27 (t, J

=8.4 Hz, 2CH), 7.24 (d, J=7.2 Hz, CH), 7.22 (d, J=7.2 Hz, 2CH), 6.56 (s, NH), 6.51 (s, 2CH), 4.35 (d, J=5.6 Hz, CH ), 4.21 (s, CH), 3.83 (d, J=4.8 Hz, 3OCH ), 3.59 (t, J=4.0 Hz, CH₂), 3.49 (q, J=4.0 Hz, CH₂), 2.41 (d, J=7.2 Hz, CH₂), 2.31-2.38 (m, CH₂), 2.29 (d, J=6.0 Hz, CH ), 2.25 (d, J=8.4 Hz, CH ), 2.14-2.20 (m, CH), 1.05 (d, J=6.4 Hz, CH

₃); [165] HR-FABMS Calcd for C H ClN O : (M⁺+l): 594.2735, Found: 594.2757.

33 40 3 5

[166]

[167] Example 4 : Preparation of

5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-N-cyclohexyl-3-methyl-5-ox opentanamide

[168]

[169] The title compound was prepared in the same manner as in Example 1, except that

4-(4-cyclohexylcarbamoyl)-3-methylbutanoic acid prepared in Preparative Example 4 was used instead of 4-(4-methylbenzylcarbamoyl)-3-methylbutanoic acid in Example 1 (white solid, yield: 73%).

[170] mp 75-76°C;

[171] ¹U NMR (CDCl ) δ 7.35 (d, J=7.2 Hz, 2CH), 7.29 (d, J=7.6 Hz, 2CH), 7.26 (t, J

=8.0 Hz, 2CH), 7.23 (d, J=7.6 Hz, CH), 7.22 (d, J=7.2 Hz, 2CH), 5.97 (s, NH), 4.21 (s, CH), 3.69-3.79 (m, CH), 3.61 (t, J=4.8 Hz, CH₂), 3.51 (q, J=4.8 Hz, CH₂), 2.43 (d, J =6.8 Hz, CH ), 2.31-2.39 (m, CH ), 2.23 (d, J=7.6 Hz, CH ), 2.16-2.20 (m, CH), 2.12 (d, J=6.0 Hz, CH₂), 1.88 (d, J=8.8 Hz, CH₂), 1.69 (d, /=12.0 Hz, CH₂), 1.59 (d, /=12.4 Hz, CH ), 1.35 (q, /=12.7 Hz, CH ), 1.14 (t, /=11.4 Hz, CH ), 1.02 (d, /=6.4 Hz, CH );

[172] HR-FABMS Calcd for C H ClN O : (M⁺+l): 496.2731, Found: 496.2724.

29 38 3 2

[173]

[174] Example 5 : Preparation of

5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-N,N,3-trimethyl-5-oxopenta namide

[175]

[176] The title compound was prepared in the same manner as in Example 1, except that

4-(dimethylcarbamoyl)-3-methylbutanoic acid prepared in Preparative Example 5 was used instead of 4-(4-methylbenzylcarbamoyl)-3-methylbutanoic acid in Example 1 (light yellow gel, yield: 8%).

[177] ¹U NMR (CDCl ) δ 7.35 (d, /=7.2 Hz, 2CH), 7.29 (d, /=7.6 Hz, 2CH), 7.26 (t, /

=8.2 Hz, 2CH), 7.23 (d, /=8.0 Hz, CH), 7.21 (d, /=7.2 Hz, 2CH), 4.21 (s, CH), 3.60-3.71 (m, CH₂), 3.48-3.59 (m, CH₂), 3.01 (s, CH₃), 2.91 (s, CH₃), 2.57; 2.53 (dd, / =5.2; 5.6 Hz, CH), 2.27-2.48 (m, 3CH ), 2.16-2.26 (m, 1/2CH ), 2.00-2.13 (m, 1/2CH ), 1.03 (d, /=6.4 Hz, CH₃);

[178] HR-FABMS Calcd for C 25 H 32 ClN 3 O 2 : (M⁺+l): 442.2261, Found: 442.2259.

[179]

[180] Example 6 : Preparation of

5-(4-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan-l-yl)-N-(4-methylbenzyl)-3-m ethyl-5-oxopentanamide

[181]

[ 182] 0.5 mmol of 4-(4-methylbenzylcarbamoyl)-3-methylbutanoic acid prepared in

Preparative Example 1, 0.55 mmol l-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan, and benzotriazol-1-yloxytripyrrolidino phosphonium hexafluorophosphate (PyBOP) were added to 3 mL of dimethylformamide (DMF) to dissolve. Then, 1.0 mmol of N,N-diisopropylethylamine (DIEA) was added thereto, and stirred at room temperature for 16 hours. 20 mL of 10% HCl was put into the reaction solution, and extracted with 30 mL of EtOAc. The organic layer was washed with 20 mL of 10% HCl, and then washed with 20 mL of a saturated NaHCO solution twice and with 20 mL of a saturated NaCl solution twice. The organic layer was collected, dried over anhydrous MgSO , and filtered under reduced pressure. The organic solvent in the filtrate was removed under reduced pressure. The residue was purified by column chromatography with a mixed solvent of EtOAc and methanol (20:1), so as to obtain the title compound (light yellow solid, yield: 75%).

[183] mp 53-54⁰C;

[184] ¹U NMR (CDCl ) δ 7.30-7.36 (m, 4CH), 7.23-7.29 (m, 3CH), 7.19-7.23 (m, 2CH),

7.17 (d, J=8.4 Hz, 2CH), 7.12 (d, J=8.0 Hz, 2CH), 6.42 (s, NH), 4.55 (d, J=4.8 Hz, CH), 4.39 (d, J=6.0 Hz, CH₂), 3.61-3.67 (m, 1/2CH₂), 3.49-3.60 (m, CH₂), 3.43-3.48 (m, 1/2CH₂), 2.53-2.68 (m, 2CH₂), 2.22-2.47 (m, 2CH₂), 2.13-2.22 (m, CH), 1.77 (q, J =5.7 Hz, CH₂), 1.07 (d, J=6.0 Hz, CH₃) 2.43 (d, J=6.8 Hz, CH₂), 2.34-2.39 (m, CH₂), 2.32 (s, CH₃), 2.28 (d, J=6.0 Hz, CH₂), 2.25 (d, J=6.4 Hz, CH₂), 2.13-2.23 (m, CH), 1.03 (d, /=6.4 Hz, CH₃);

[185] HR-FABMS Calcd for C 32 H 38 ClN 3 O 2 : ( ^VM⁺+l) ': 532.2731, Found: 532.2720.

[186]

[187] Example 7 : Preparation of

5-(4-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan-l-yl)-N-(4-methoxybenzyl)-3- methyl-5-oxopentanamide

[188]

[189] The title compound was prepared in the same manner as in Example 6, except that

4-(4-methoxybenzylcarbamoyl)-3-methylbutanoic acid prepared in Preparative Example 2 was used instead of 4-(4-methylbenzylcarbamoyl)-3-methylbutanoic acid in Example 6 (brown solid, yield: 66%).

[190] mp 52-53⁰C;

[191] ¹U NMR (CDCl ) δ 7.30-7.36 (m, 4CH), 7.24-7.29 (m, 3CH), 7.18-7.23 (m, 2CH),

6.84 (d, J=6.8 Hz, 2CH), 6.43 (s, NH), 4.55 (d, J=4.4 Hz, CH), 4.36 (d, J=5.6 Hz, CH₂ ), 3.79 (s, OCH ), 3.61-3.67 (m, 1/2CH ), 3.53-3.58 (m, CH ), 3.43-3.48 (m, 1/2CH ), 2.54-2.68 (m, 2CH ), 2.22-2.47 (m, 2CH ), 2.13-2.21 (m, CH), 1.77 (q, /=6.1 Hz, CH ), 1.07 (d, /=6.0 Hz, CH );

[192] HR-FABMS Calcd for C 32 H 38 ClN 3 O 3 : (M⁺+l): 548.2680, Found: 548.2690.

[193]

[194] Example 8 : Preparation of

5-(4-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan-l-yl)-N-(3,4,5-trimethoxyben zyl)-3-methyl-5-oxopentanamide

[195]

[196] The title compound was prepared in the same manner as in Example 6, except that

4-(3,4,5-trimethoxybenzylcarbamoyl)-3-methylbutanoic acid prepared in Preparative Example 3 was used instead of 4-(4-methylbenzylcarbamoyl)-3-methylbutanoic acid in Example 6 (white solid, yield: 46%).

[197] mp 60-61⁰C;

[198] ¹U NMR (CDCl ) δ 7.30-7.35 (m, 4CH), 7.25-7.29 (m, 3CH), 7.18-7.24 (m, 2CH),

6.66 (s, NH), 6.53 (s, 2CH), 4.55 (d, /=4.0 Hz, CH), 4.37 (d, /=6.0 Hz, CH₂), 3.83 (d, / =5.2 Hz, 3OCH ), 3.61-3.67 (m, 1/2CH ), 3.55-3.59 (m, CH ), 3.46 (t, /=4.6 Hz, 1/2CH ), 2.53-2.69 (m, 2CH ), 2.26-2.47 (m, 2CH ), 2.16-2.22 (m, CH), 1.77 (q, /=5.7 Hz, CH ), 1.08 (d, /=5.8 Hz, CH );

[199] HR-FABMS Calcd for C 34 H 42 ClN 3 O 5 : (M⁺+l): 608.2891, Found: 608.2901.

[200]

[201] Example 9 : Preparation of

5-(4-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan-l-yl)-N-cyclohexyl-3-methyl-

5-oxopentanamide

[202]

[203] The title compound was prepared in the same manner as in Example 6, except that 4-(4-cyclohexylcarbamoyl)-3-methylbutanoic acid prepared in Preparative Example 4 was used instead of 4-(4-methylbenzylcarbamoyl)-3-methylbutanoic acid in Example 6 (white solid, yield: 60%).

[204] mp 62-63⁰C;

[205] ¹U NMR (CDCl ) δ 7.31-7.35 (m, 4CH), 7.25-7.30 (m, 3CH), 7.18-7.24 (m, 2CH),

5.06 (s, NH), 4.55 (s, CH), 3.71-3.82 (m, CH), 3.63-3.70 (m, 1/2CH₂), 3.59 (t, J=6.0 Hz, CH ), 3.46-3.50 (m, 1/2CH ), 2.61-2.68 (m, CH ), 2.55-2.60 (m, CH ), 2.35-2.47 (m, CH₂), 2.22-2.33 (m, CH), 2.09-2.15 (m, CH₂), 1.89 (d, /=12.0 Hz, CH₂), 1.78 (q, / =6.0 Hz, CH ), 1.70 (d, /=13.6 Hz, CH ), 1.60 (d, /=12.4 Hz, CH ), 1.36 (q, /=12.5 Hz, CH₂), 1.15 (t, /=10.2 Hz, CH₂), 1.05 (d, /=5.8 Hz, CH₃);

[206] HR-FABMS Calcd for C H ClN O : (M⁺+l): 510.2887, Found: 510.2864.

30 40 3 2

[207]

[208] Preparative Example 6 : Preparation of N-

((t-butyloxy)carbonyl)-N'-(dimethyl)iminodiacetic acid monoamide

[209] 2 mmol of N-((t-butyloxy)carbonyl)iminodiacetic acid and l-(3-dimethylaminopropyl)-3-ethyl-carbodiamide HCl (EDCI) were added to 3 mL of dimethylformamide (DMF) to dissolve, and stirred at room temperature for 1 hour. Then, 2 mmol of dimethylamine was added thereto, and stirred at room temperature for 20 hours. 20 mL of 10% HCl was put into the reaction solution, and extracted with 30 mL of EtOAc. The organic layer was washed with 20 mL of 10% HCl, and then washed with 20 mL of a saturated NaCl solution twice. The organic layer was collected, dried over anhydrous MgSO , and filtered under reduced pressure. The organic solvent in the filtrate was removed under reduced pressure. The residue was re- crystallized from a mixed solvent of EtOAc and methanol, so as to obtain the title compound (white solid, yield: 33%).

[210] mp 144-145⁰C;

[211] ¹H NMR (CDCl₃) δ 4.18 (s, CH₂), 3.89 (s, CH₂), 3.05 (s, 2 CH₃), 1.47 (s, 3CH₃);

[212] HR-FABMS Calcd for C H N O (M⁺+l): 261.1450, Found: 261.1444.

11 20 2 5

[213]

[214] Example 10 : Preparation of t-butyl (dimethylcarbamoyl)methyl

2-(4-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan-l-yl)-2-oxoethylcarbamate

[215]

[216] 0.2 mmol of the compound prepared in Preparative Example 6, 0.22 mmol of l-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan, and PyBOP were added to 3 mL of DMF to dissolve. Then, 0.4 mmol of DIEA was added thereto, and stirred at room temperature for 16 hours. 20 mL of 10% HCl was put into the reaction solution, and extracted with 30 mL of EtOAc. The organic layer was washed with 20 mL of 10% HCl, and then washed with 20 mL of a saturated NaHCO solution twice and with 20 mL of a saturated NaCl solution twice. The organic layer was collected, dried over anhydrous MgSO , and filtered under reduced pressure. The organic solvent in the

4 filtrate was removed under reduced pressure. The residue was purified by column chromatography with a mixed solvent of EtOAc and methanol (20: 1), so as to obtain the title compound (light yellow gel, yield: 24%).

[217] ¹U NMR (CDCl ) δ 7.30-7.38 (m, 3CH), 7.22-7.29 (m, 4CH), 7.16-7.22 (m, 2CH),

4.58 (s, CH), 4.18-4.27 (m, CH ), 4.09-4.16 (m, CH ), 3.56-3.68 (m, CH ), 3.38-3.55 (m, CH₂), 2.99 (s, CH₃), 2.93 (s, CH₃), 2.64 (t, J=5.2 Hz, CH₂), 2.58 (t, J=5.6 Hz, CH₂), 1.73-1.85 (m, CH₂), 1.43 (s, 3CH₃);

[218] HR-FABMS Calcd for C H ClN O : (M⁺+l): 543.2738, Found: 543.2741.

29 39 4 4

[219]

[220] Example 11 : Preparation of

2-(2-(4-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan-l-yl)-2-oxoethylamino)-N,

N-dimethylacetamide

[221]

[222] 1 ml of 4M HCl-dioxane and 1 ml of CHCl were added to 0.05 mmol of the compound prepared in Example 10, and stirred at room temperature for 3 hours. The reaction solution was evaporated under reduced pressure. The residue was purified by column chromatography with a mixed solvent of EtOAc and methanol (2: 1), so as to obtain the title compound (light yellow gel, yield: 28%).

[223] ¹U NMR (CDCl ) δ 7.31-7.37 (m, 4CH), 7.26-7.30 (m, 2CH), 7.17-7.25 (m, 3CH),

4.56 (s, CH), 3.67 (t, J=6.2 Hz, 1/2CH ), 3.62 (t, J=5.6 Hz, 1/2CH ), 3.48-3.56 (m, 5/2CH₂), 3.39 (t, J=5.4 Hz, 1/2CH₂), 2.96 (s, 2CH₃), 2.66 (t, J=5.0 Hz, OT), 2.57 (t, J =5.4 Hz, CH ), 1.73-1.84 (m, CH );

[224] HR-FABMS Calcd for C 24 H 31 ClN 4 O 2 : (M⁺+Na): 465.2033, Found: 465.2040.

[225]

[226] Preparative Example 7 : Preparation of 3-(4-methylbenzylcarbamoyl)-propanoic acid

[227] 2 mmol of succinic anhydride was added to 3 mL of dimethylformamide (DMF) to dissolve. 2 mmol of 4-methylbenzylamine was added thereto, and stirred at room temperature for 20 hours. 20 mL of 10% HCl was put into the reaction solution, and extracted with 30 mL of EtOAc. The organic layer was washed with 20 mL of 10% HCl, and then washed with 20 mL of a saturated NaCl solution twice. The organic layer was collected, dried over anhydrous MgSO , and filtered under reduced pressure. The organic solvent in the filtrate was removed under reduced pressure. The residue was recrystallized from a mixed solvent of EtOAc and methanol, so as to obtain the title compound (white solid, yield: 77%).

[228] mp 146.7-149.2⁰C;

[229] ¹U NMR (CDCl ) δ 7.18-7.14 (m, 4CH), 5.90 (br s, NH), 4.42 (s, CH ), 2.74 (t, J

=6.0 Hz, CH ), 2.55 (t, J=6.0 Hz, CH ) 2.34 (s, CH );

[230] HR-FABMS Calcd for C H NO (M⁺+ 1): 222.1130, Found: 222.1127.

12 16 3

[231]

[232] Preparative Example 8 : Preparation of

3-(4-methoxybenzylcarbamoyl)-propanoic acid

[233] The title compound was prepared in the same manner as in Preparative Example 7, except that 4-methoxybenzylamine was used instead of 4-methylbenzylamine in Preparative Example 7 (light yellow solid, yield: 56%).

[234] mp 156.7-162.9⁰C;

[235] ¹U NMR (CDCl ) δ 7.24-7.19 (m, 4CH), 6.99-6.89 (m, 2H), 5.87 (br s, NH), 4.40

(s, CH₂), 3.80 (s, OCH3), 2.73 (t, J=6.4 Hz, CH₂), 2.56 (t, J=6.4Hz, CH₂);

[236] HR-FABMS Calcd for C H NO (M⁺+l): 238.1079, Found: 238.1076.

12 16 4

[237]

[238] Preparative Example 9 : Preparation of

3-(3,4,5-trimethoxybenzylcarbamoyl)-propanoic acid

[239] The title compound was prepared in the same manner as in Preparative Example 7, except that 3,4,5-trimethoxybenzylamine was used instead of 4-methylbenzylamine in Preparative Example 7 (light yellow solid, yield: 18%).

[240] mp 121.8-126.9⁰C;

[241] ¹U NMR (CDCl ) δ 6.49 (s, 2CH), 4.38 (s, CH ), 3.85 (s, 3OCH ), 2.74 (t, J

=6.8Hz, CH₂), 2.55(t, J=6.8 Hz, CH₂);

[242] HR-FABMS Calcd for C 14 H 20 NO 6 ( ^VM⁺+l) ': 298.1291, Found: 298.1286.

[243]

[244] Preparative Example 10 : Preparation of 3-(cyclohexylcarbamoyl)-propanoic acid

[245] The title compound was prepared in the same manner as in Preparative Example 7, except that cyclohexylamine was used instead of 4-methylbenzylamine in Preparative

Example 7 (white solid, yield: 60%). [246] mp 169.4-173.6⁰C;

[247] ¹U NMR (CDCl ) δ 5.58(s, NH), 3.81-3.74 (m, CH), 2.69 (t, J=6.4 Hz, CH ), 2.51

(t, J=6.4 Hz, CH ), 1.95-1.90 (m, CH ), 1.75-1.60 (m, 2CH ), 1.42-1.13 (m, 2CH ); [248] HR-FABMS Calcd for C 10 H 17 NO 3 (M⁺+Na): 222.1106, Found: 222.1103.

[249]

[250] Example 12 : Preparation of N-

(4-methylbenzyl)-4-oxo-4-((4-chlorobenzhydryl)piperazin-l-yl)butanamide

[251]

[252] 0.5 mmol of 3-(4-methylbenzylcarbamoyl)-propanoic acid prepared in Preparative

Example 7, 0.55 mmol of l-(4-chlorobenzhydryl)piperazine, and benzotriazol- 1-yloxytripyrrolidino phosphonium hexafluorophosphate (PyBOP) were added to 3 mL of dimethylformamide (DMF) to dissolve. Then, 1.0 mmol of

N,N-diisopropylethylamine (DIEA) was added thereto, and stirred at room temperature for 16 hours. 20 mL of 10% HCl was put into the reaction solution, and extracted with 30 mL of EtOAc. The organic layer was washed with 20 mL of 10% HCl, and then washed with 20 mL of a saturated NaHCO solution twice and with 20 mL of a saturated NaCl solution twice. The organic layer was collected, dried over anhydrous MgSO , and filtered under reduced pressure. The organic solvent in the filtrate was removed under reduced pressure. The residue was purified by column chromatography with a mixed solvent of EtOAc and methanol (20:1), so as to obtain the title compound (light yellow solid, yield: 60%).

[253] mp 74.5-80.3⁰C;

[254] ¹U NMR (CDCl 3 ) δ 7.37-7.34 (m, 4CH), 7.32-7.13 (m, 5CH), 7.15-7.11 (t, J

=4.4Hz, CH ), 3.46 (t, J=4.4Hz, CH ), 2.64 (t, J=6.4Hz, CH ), 2.53 (t, J=6.4Hz, CH ), 2.36-2.32 (m, 2CH₂), 2.31(s, CH₃);

[255] HR-FABMS Calcd for C 29 H 33 ClN 3 O 2 : (M⁺+l): 490.2261, Found: 490.2249.

[256]

[257] Example 13 : Preparation of N-

(4-methoxybenzyl)-4-oxo-4-((4-chlorobenzhydryl)piperazin-l-yl)butanamide

[258]

[259] The title compound was prepared in the same manner as in Example 12, except that

3-(4-methoxybenzylcarbamoyl)-propanoic acid prepared in Preparative Example 8 was used instead of 3-(4-methylbenzylcarbamoyl)-3-propanoic acid in Example 12 (light yellow solid, yield: 44%).

[260] mp 114.0-122.1⁰C;

[261] ¹U NMR (CDCl ) δ 7.36- 7.32 (m, 4CH), 7.29-7.19 (m, 5CH), 7.17-7.15 (m, 2CH

), 6.82-6.78 (m, 2CH), 4.31(s, CH ), 4.19 (s, CH), 3.74 (s, OCH ), 3.59-3.50 (m, CH ), 3.44-3.31 (m, CH₂), 2.61 (t, J=6.4 Hz, CH₂), 2.94 (t, J=6.4 Hz, CH₂), 2.34-2.33 (m, CH ), 2.30-2.28 (m, CH );

[262] HR-FABMS Calcd for C 29 H 33 ClN 3 O 3 : (M⁺+l): 506.2210, Found: 5506.2196.

[263]

[264] Example 14 : Preparation of N-

(3,4,5-trimethoxybenzyl)-4-oxo-4-((4-chlorobenzhydryl)piperazin-l-yl)butanamid e

[265]

[266] The title compound was prepared in the same manner as in Example 12, except that

3-(3,4,5-trimethoxybenzylcarbamoyl)-propanoic acid prepared in Preparative Example 9 was used instead of 3-(4-methylbenzylcarbamoyl)-3-propanoic acid in Example 12 (white solid, yield: 20%).

[267] mp 70.6-73.6⁰C;

[268] ¹U NMR (CDCl ) δ 7.37-7.3 l(m, 4CH), 7.30-7.25(m, 5CH), 6.50(s, 2CH), 4.35(s,

CH₂), 4.21 (s, CH), 3.84 (s, 3OCH₃), 3.56-3.46 (m, 2CH₂), 2.66 (t, J=6.4Hz, CH₂), 2.54 (t, J=6.4Hz, CH₂), 2.38-2.27 (m, 2CH₂);

[269] HR-FABMS Calcd for C 31 H 37 ClN 3 O 5 : (M⁺+l): 566.2422, Found: 566.2415.

[270]

[271] Example 15 : Preparation of N-

(cyclohexyl)-4-oxo-4-((4-chlorobenzhydryl)piperazin-l-yl)butanamide

[272]

[273] The title compound was prepared in the same manner as in Example 12, except that

3-(cyclohexylcarbamoyl)-propanoic acid prepared in Preparative Example 10 was used instead of 3-(4-methylbenzylcarbamoyl)-3-propanoic acid in Example 12 (yellow solid, yield: 52%).

[274] mp 52.6-60.5⁰C;

[275] ¹U NMR (CDCl ) δ 7.36-7.34 (m, 4CH), 7.30-7.19 (m, 5CH), 5.34 (s, NH), 4.21 (s,

CH), 3.60-3.55 (m, CH ), 3.49-3.46 (m, CH ), 2.61 (t, J=6.8Hz, CH ), 2.46 (t, J=6.8Hz, CH₂), 2.35-2.33 (m, 2CH₂);

[276] HR-FABMS Calcd for C 27 H 35 ClN 3 O 2 : ( ^VM⁺+l) ': 468.2418, Found: 468.2407.

[211]

[278] Example 16 : Preparation of N-

(4-methylbenzyl)-4-oxo-4-((4-chlorobenzhydryl)-l,4-diazepan-l-yl)butanamide

[279]

[280] 0.5 mmol of 3-(4-methylbenzylcarbamoyl)-propanoic acid prepared in Preparative

Example 7, 0.55 mmol of l-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan, and PyBOP were added to 3 mL of DMF to dissolve. Then, 1.0 mmol of N,N-diisopropylethylamine (DIEA) was added thereto, and stirred at room temperature for 16 hours. 20 mL of 10% HCl was put into the reaction solution, and extracted with 30 mL of EtOAc. The organic layer was washed with 20 mL of 10% HCl, and then washed with 20 mL of a saturated NaHCO solution twice and with 20 mL of a

3 saturated NaCl solution twice. The organic layer was collected, dried over anhydrous MgSO , and filtered under reduced pressure. The organic solvent in the filtrate was removed under reduced pressure. The residue was purified by column chromatography with a mixed solvent of EtOAc and methanol (20:1), so as to obtain the title compound (light yellow solid, yield: 58%).

[281] mp 71.5-73.8°C;

[282] ¹U NMR (CDCl₃); δ 7.36- 7.34 (m,4CH), 7.23-7.18 (m, 5CH), 7.17-7.11 (m, 4CH),

6.34 (s, NH), 4.55 (s, CH), 4.39 (s, CH ), 3.66-3.45 (m, 2CH ), 2.72-2.66 (m, 2CH ), 2.67-2.53 (m, 4CH), 2.32 (s, CH₃), 1.82- 1.73 (m, CH₂); [283] HR-FABMS Calcd for C 30 H 35 ClN 3 O 2 : (M⁺+l): 504.2418, Found: 504.2393.

[284]

[285] Example 17 : Preparation of N-

(4-methoxybenzyl)-4-oxo-4-((4-chlorobenzhydryl)-l,4-diazepan-l-yl)butanamide

[286]

[287] The title compound was prepared in the same manner as in Example 16, except that

3-(4-methoxybenzylcarbamoyl)-propanoic acid prepared in Preparative Example 8 was used instead of 3-(4-methylbenzylcarbamoyl)-3-propanoic acid in Example 16 (light yellow solid, yield: 21%).

[288] mp 54.2-60.4⁰C;

[289] ¹U NMR (CDCl ) δ 7.36-7.31 (m, 4CH), 7.29-7.20 (m, 5CH), 7.19-7.17 (m, 2CH),

6.84-6.81 (m, 2CH), 6.85 (s, NH), 4.66 (d, J=6.8Hz, CH), 4.35 (d, J=5.6Hz, CTT), 3.77 (s, OCH₃), 3.63-3.55 (m, CH₂), 3.54-3.41 (m, CH₂), 2.70-2.52 (m, 4CH₂), 1.81-1.71 (m, CH₂);

[290] HR-FABMS Calcd for C H ClN O : (M⁺+l): 520.2367, Found: 520.2346.

30 35 3 3

[291]

[292] Example 18 : Preparation of N-

(3,4,5-trimethoxybenzyl)-4-oxo-4-((4-chlorobenzhydryl)-l,4-diazepan-l-yl)butana mide

[293]

[294] The title compound was prepared in the same manner as in Example 16, except that

3-(3,4,5-trimethoxybenzylcarbamoyl)-propanoic acid prepared in Preparative Example 9 was used instead of 3-(4-methylbenzylcarbamoyl)-3-propanoic acid in Example 16 (light yellow solid, yield: 41%).

[295] mp 78.1-85.7°C;

[296] ¹U NMR (CDCl ) δ 7.37-7.33 (m, 4cH), 7.29-7.17 (m, 5CH), 6.52 (s, 2CH), 4.55

(d, J=6.4Hz, CH), 4.37 (d, J=5.6Hz, CH₂), 3.84 (s, 3OCH₃), 3.65-3.62 (m, 1/2CH₂), 3.59-3.55 (m, CTT), 3.48-3.45 (m,l/2CH ), 2.74-2.53 (m, 4CH₂), 1.83-1.72 (m,CH );

[297] HR-FABMS Calcd for C 32 H 39 ClN 3 O 5 : (M⁺+l): 580.2578, Found: 580.2588.

[298] [299] Example 19 : Preparation of N-

(cyclohexyl)-4-oxo-4-((4-chlorobenzhydryl)-l,4-diazepan-l-yl)butanamide

[300]

[301] The title compound was prepared in the same manner as in Example 16, except that

3-(cyclohexylcarbamoyl)-propanoic acid in Preparative Example 10 was used instead of 3-(4-methylbenzylcarbamoyl)-3-propanoic acid in Example 16 (white solid, yield: 55%).

[302] mp 58.4-70.6⁰C;

[303] ¹H NMR (CDCy δ 7.36-7.32 (m, 4CH), 7.29-7.18 (m, 5CH), 6.16 (s, NH), 4.56 (s,

CH), 3.76-3.69 (m, CH), 3.65 (t, J=5.6Hz, 1/2CH₂), 3.58 (t, J=5.6Hz, CH₂), 3.45 (t, J =5.6Hz, 1/2CH ), 2.69-2.49 (m, 4CH ), 1.89-1.85 (m, CH ), 1.81-1.74 (m, CH ), 1.71-1.65 (m, CH₂), 1.61-1.56 (m, 1/2CH₂), 1.38-1.30 (m, CH₂), 1.19-1.06 (m, CH₂);

[304] HR-FABMS Calcd for C H ClN O : (M⁺+l): 482.2574, Found: 482.2551.

28 37 3 2

[305]

[306] Example 20 : Preparation of N-

(3,4,5-trimethoxybenzyl)-N'-l-((4-chlorobenzhydryl)-l,4-diazepan-l-yl)iminodiac etic acid diamide

[307]

[308] 0.5 mmol of N-((t-butyloxy)carbonyl)-N'-(3,4,5-trimethoxybenzyl)iminodiacetic acid monoamide, 0.55 mmol of l-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan, and PyBOP were added to 3 mL of DMF to dissolve. Then, 1.0 mmol of N,N-diisopropylethylamine (DIEA) was added thereto, and stirred at room temperature for 16 hours. 20 mL of 10% HCl was put into the reaction solution, and extracted with 30 mL of EtOAc. The organic layer was washed with 20 mL of 10% HCl, and then washed with 20 mL of a saturated NaHCO 3 solution twice and with 20 mL of a saturated NaCl solution twice. The organic layer was collected, dried over anhydrous MgSO 4 , and filtered under reduced pressure. The organic solvent in the filtrate was removed under reduced pressure. The residue was purified by column chromatography with a mixed solvent of EtOAc and methanol (20: 1), so as to obtain N- ((t-butyloxy)carbonyl)-N'-(3,4,5-trimethoxybenzyl)-N"-l-((4-chlorophenyl)(phenyl)me thyl)-l,4-diazepan iminodiacetic acid diamide. Chloroform (1 mL) and 4 M HCl- dioxane (1 mL) were added to the obtained compound, and left to stand for 3 hours. The solvent was removed under reduced pressure, so as to obtain the title compound (light yellow solid, yield: 31%).

[309] mp 78.4-80.7⁰C;

[310] ¹U NMR (CDCl ); δ 7.47-7.43 (m, 4CH), 129-1 Al (m, 5CH), 6.02 (s, 2CH),

4.89(s, CH), 4.45 (d, J=6.4Hz, CH₂), 3.46 (m, 2CH₂), 3.74 (s, 3OCH₃), 3.65-3.62 (m, 1/2CH ), 3.59-3.50 (m, CH ), 3.48-3.46 (m,l/2CH ), 2.75-2.50 (m, 4CH ), 1.80-1.72 (m,CH₂);

[311] HR-FABMS Calcd for C 32 H 39 ClN 3 O 5 : (M⁺+l): 595.2609, Found: 595.2613.

[312]

[313] Preparative Example 11 : Preparation of

(Z)-3-(4-methylbenzylcarbamoyl)acrylic acid

[314] 1 mmol of maleic acid was added to 3 mL of dimethylformamide (DMF) to dissolve. Then, 1 mmol of l-(3-dimethylaminopropyl)-3-ethyl-carbodiamide HCl (EDCI) was added thereto, and stirred at room temperature for 1 hour. After 1 hour, 4-methylbenzylamine was added thereto, and stirred at room temperature for 16 hours. 30 mL of 10% HCl was put into the reaction solution, and extracted with 50 mL of EtOAc. The organic layer was washed with 30 mL of 10% HCl, and then washed with 20 mL of a saturated NaCl solution twice. The organic layer was collected, dried over anhydrous MgSO , and filtered under reduced pressure. The organic solvent in the filtrate was removed under reduced pressure. The residue was recrystallized from a mixed solvent of EtOAc and methanol, so as to obtain the title compound (white solid, yield: 73%).

[315] mp 143-144⁰C;

[316] ¹U NMR (CDCl , 400MHz) δ 2.350(s, 3H), 4.225(s, 2H), 6.077(d, IH, J=12.8Hz),

6.325(d, IH, J=12.8Hz), 6.923~6.945(m, 4H)

[317]

[318] Preparative Example 12 : Preparation of

(Z)-3-(4-methoxybenzylcarbamoyl)acrylic acid

[319] The title compound was prepared in the same manner as in Preparative Example 11, except that 4-methoxybenzylamine was used instead of 4-methylbenzylamine in Preparative Example 11 (white solid, yield: 72%).

[320] mp 138-146⁰C;

[321] ¹U NMR (CDCl₃ 400MHz) δ 3.810(s, 3H), 4.395(s, 2H), 6.051(d, IH, J=12.8Hz),

6.402(d, IH, J=12.8Hz), 6.553~6.588(m, 2H), 6.766~6.800(m, 2H)

[322] [323] Preparative Example 13 : Preparation of

(Z)-3-(4-phenylbutan-2-yl-carbamoyl)acrylic acid

[324] The title compound was prepared in the same manner as in Preparative Example 11, except that l-methyl-3-phenylpropylamine was used instead of 4-methylbenzylamine in Preparative Example 11 (white solid, yield: 91%). [325] mp lO8-115°C;

[326] ¹U NMR (CDCl , 400MHz) δ 1.302(d, 3H, J= 6.8Hz), 1.838-1.402 (m, 2H),

2.602~2.630(m, 2H), 3.762~3.796(m, IH), 6.510(d, IH, J=12.8Hz), 6.75(d, IH,

J=12.8Hz), 7.08~7.21(m, 5H) [327] [328] Example 21 : Preparation of

(Z)-N-(4-methylbenzyl)-4-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-4-o xo-but-2-enamide [329]

[330] 0.5 mmol of (Z)-3-(4-methylbenzylcarbamoyl)acrylic acid prepared in Preparative

Example 11, 0.55 mmol of l-(4-chlorobenzhydryl)piperazine, and benzotriazol- 1-yloxytripyrrolidino phosphonium hexafluorophosphate (PyBOP) were added to 3 mL of DMF to dissolve. Then, 1.0 mmol of N,N-diisopropylethylamine (DIEA) was added thereto, and stirred at room temperature for 16 hours. 20 mL of 10% HCl was put into the reaction solution, and extracted with 30 mL of EtOAc. The organic layer was washed with 20 mL of 10% HCl, and then washed with 20 mL of a saturated NaHCO solution twice and with 20 mL of a saturated NaCl solution twice. The organic layer was collected, dried over anhydrous MgSO , and filtered under reduced pressure. The organic solvent in the filtrate was removed under reduced pressure. The residue was purified by column chromatography with a mixed solvent of EtOAc and methanol (20:1), so as to obtain the title compound (light yellow solid, yield: 81%).

[331] mp 76.1-78.0°C;

[332] ¹U NMR (CDCl 400MHz) δ 2.310~2.232(m, 4H), 2.337(s, 3H), 3.441-3.467 (m,

2H), 3.591~3.614(m, 2H), 4.204(s, IH), 4.425(d, 2H, J=5.6Hz), 6.076(d, IH, J=12.8Hz), 6.328(d, IH, J=12.8Hz), 7.119~7.169(m, 4H), 7.189~7.234(m, 2H), 7.290~7.308(m, 2H), 7.330~7.360(m, 5H);

[333] FABHRMS (m/z): 488.2105 (M⁺+l, requires C H31O N Cl:488.2027).

[334]

[335] Example 22 : Preparation of (Z)-N-(4-methoxybenzyl)-4-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-4- oxo-but-2-enamide

[336]

[337] The title compound was prepared in the same manner as in Example 21, except that

(Z)-3-(4-methoxybenzylcarbamoyl)acrylic acid prepared in Preparative Example 12 was used instead of (Z)-3-(4-methylbenzylcarbamoyl)acrylic acid in Example 21 (light yellow solid, yield: 80%).

[338] mp 77-78.O⁰C;

[339] ¹U NMR (CDCl 400MHz) δ 2.290~2.329(m, 4H), 3.429- 3.454(m, 2H),

3.578~3.619(m, 2H), 3.801(s, 3H), 4.186(s,lH), 4.396(d, 2H, J=6.0Hz), 6.072(d, IH, J=12.8Hz), 6.322(d, IH, J=12.8Hz), 6.850~6.872(m, 2H), 7.196~7.250(m, 4H), 7.284~7.305(m, 2H), 7.330~7.360(m,4H);

[340] FABHRMS (m/z): 504.2054(M⁺+l, requires C H N O Cl: 504.1976).

[341]

[342] Example 23 : Preparation of

(Z)-4-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-4-oxo-N-(4-phenylbutan -2-yl)-but-2-enamide

[343]

[344] The title compound was prepared in the same manner as in Example 21, except that

(Z)-3-(4-phenylbutan-2-yl-carbamoyl)acrylic acid prepared in Preparative Example 13 was used instead of (Z)-3-(4-methylbenzylcarbamoyl)acrylic acid in Example 21 (light yellow solid, yield: 89%).

[345] mp 154-157⁰C;

[346] ¹U NMR (CDCl₃ 400MHz) δ 1.184(d, J=6.8Hz), 1.741~1.824(m, 2H),

2.286~2.386(m, 4H), 2.602~2.633(m, 2H), 3.463~3.489(m, 2H), 3.620~3.694(m, 2H), 4.026~4.063(m, IH), 4.139(s, IH), 6.059(d, IH, J=12.8Hz), 6.305(d, IH, J=12.8Hz), 7.176~7.196(m, 2H), 7.210~7.324(m, 12H);

[347] FABHRMS (m/z):516.2418 (M⁺+l, requires C 31 H 34 N 3 O 2 Cl: 516.2340).

[348]

[349] Example 24 : Preparation of (Z)-N-(4-methylbenzyl)-4-(4-(bis(4-fluorophenyl)methyl)piperazin-l-yl)-4-oxo-but -2-enamide

[350]

[351] 0.5 mmol of (Z)-3-(4-methylbenzylcarbamoyl)acrylic acid prepared in Preparative

Example 11, 0.55 mmol of l-(bis(4-fluorophenyl)methyl)piperazine, and benzotriazol- 1-yloxytripyrrolidino phosphonium hexafluorophosphate (PyBOP) were added to 3 mL of DMF to dissolve. Then, 1.0 mmol of N,N-diisopropylethylamine (DIEA) was added thereto, and stirred at room temperature for 16 hours. 20 mL of 10% HCl was put into the reaction solution, and extracted with 30 mL of EtOAc. The organic layer was washed with 20 mL of 10% HCl, and then washed with 20 mL of a saturated NaHCO solution twice and with 20 mL of a saturated NaCl solution twice. The organic layer was collected, dried over anhydrous MgSO , and filtered under reduced pressure. The organic solvent in the filtrate was removed under reduced pressure. The residue was purified by column chromatography with a mixed solvent of EtOAc and methanol (20:1), so as to obtain the title compound (light red solid, yield: 67%).

[352] mp 68.4-75.O⁰C;

[353] ¹U NMR (CDCl 400MHz) δ 2.296~2.305(m,4H), 2.332(s, H), 3.438~3.487(m,

2H), 3.589~3.614(m, 2H), 4.213(s, IH), 4.421(d, 2H, J=6.0Hz), 6.070(d, IH, J=12.8Hz), 6.319(d, IH, J=12.8Hz), 6.963~7.012(m, 4H), 7.114-7.184(m, 4H), 7.308~7.344(m, 4H);

[354] FABHRMS (m/z): 490.2306 (M⁺+l, requires C 29 H 30O 2N 3 F 2 : 490.2228).

[355]

[356] Example 25 : Preparation of

(Z)-N-(4-methoxybenzyl)-4-(4-(bis(4-fluorophenyl)methyl)piperazin-l-yl)-4-oxo-b ut-2-enamide

[357]

[358] The title compound was prepared in the same manner as in Example 24, except that

(Z)-3-(4-methoxybenzylcarbamoyl)acrylic acid prepared in Preparative Example 12 was used instead of (Z)-3-(4-methylbenzylcarbamoyl)acrylic acid in Example 24 (light yellow solid, yield: 73%). [359] mp 72.5-75.O⁰C;

[360] ¹U NMR (CDCl 400MHz) δ 2.292(br s, 4H), 3.342(br s, 2H), 3.587(br s, 2H),

3.787(s, 3H0, 4.192(s, IH), 4.38 l(d, 2H, J=5.6Hz), 6.058(d, IH, J=12.8Hz), 6.300(d,

IH, J=12.8HZ), 6.837~6.858(m, 2HO, 6.957~6.999(m, 4H), 7.176~7.220(m, 2H),

7.307~7.340(m, 4H);

[361] FABHRMS (m/z):506.2255 (M⁺+l, requires C H N O F : 506.2177). ⁿ 29 30 3 3 2 [362] [363] Example 26 : Preparation of

(Z)-4-(4-(bis(4-fluorophenyl)methyl)piperazin-l-yl)-4-oxo-N-(4-phenylbutan-2-yl) -but-2-enamide

[364]

[365] The title compound was prepared in the same manner as in Example 24, except that (Z)-3-(4-phenylbutan-2-yl-carbamoyl)acrylic acid prepared in Preparative Example 13 was used instead of (Z)-3-(4-methylbenzylcarbamoyl)acrylic acid in Example 24 (light yellow solid, yield: 68%).

[366] mp 148-152⁰C; [367] ¹U NMR (CDCl 400MHz) δ 1.183(d, 3H, J=6.4Hz), 1.741-1.8O8(m, 2H), 2.258~2.378(m, 4H), 2.601~2.661(m, 2H), 3.461~3.488(m, 2H), 3.604~3.686(m, 2H), 4.027~4.064(m, IH), 4.143(s, IH), 6.059(d, IH, J=12.8Hz), 6.304(d, IH, J=12.8Hz), 6.937~6.996(m, 4H), 7.176~7.196(m, 4H), 7.248~7.303(m, 5H);

[368] FABHRMS (m/z):518.2619 (M⁺+l, requires C H N O F : 518.2541). ⁿ 31 34 3 2 2 [369] [370] Example 27 : Preparation of

2-(2-(4-(bis(4-fluorophenyl)methyl)piperazin-l-yl)-2-oxoethylamino)-N-(4-phenyl butan-2-yl)acetamide

[371]

[372] 0.5 mmol of N-((t-butyloxy)carbonyl)-N'-(l-methyl-3-phenylpropyl)iminodiacetic acid monoamide, 0.55 mmol of l-(bis(4-fluorophenyl)methyl)piperazine, and PyBOP were added to 3 mL of DMF to dissolve. Then, 1.0 mmol of

N,N-diisopropylethylamine (DIEA) was added thereto, and stirred at room temperature for 16 hours. 20 mL of 10% HCl was put into the reaction solution, and extracted with 30 mL of EtOAc. The organic layer was washed with 20 mL of 10% HCl, and then washed with 20 mL of a saturated NaHCO solution twice and with 20 mL of a saturated NaCl solution twice. The organic layer was collected, dried over anhydrous MgSO , and filtered under reduced pressure. The organic solvent in the filtrate was removed under reduced pressure. The residue was purified by column chromatography with a mixed solvent of EtOAc and methanol (20: 1), so as to obtain N- ((t-butyloxy)carbonyl)-N'-(l-methyl-3-phenylpropyl)-N"-l-(bis(4-fluorophenyl)methyl )piperazine iminodiacetic acid diamide. Chloroform (1 mL) and 4 M HCl-dioxane (1 mL) were added to the obtained compound, and left to stand for 3 hours. The solvent was removed under reduced pressure, so as to obtain the title compound (white solid, yield: 56%).

[373] mp 148-152⁰C;

[374] ¹U NMR (CDCl 400MHz) δ 1.147(d, 3H, J=6.4Hz), 1.605-1.846(m, 2H),

2.575-2.716(m, 2H), 3.163~3.431(m, 4H), 3.776~3.994(m, 4H), 4.052~4.214(m, 2H), 4.326~4.499(m, 2H), 5.486(br s, IH), 6.010(s, IH), 7.046- 7.247 (m, 8H), 8.173(br s, 4H), 8.520~8.650(m, IH);

[375] FABHRMS (m/z):535.2885 (M⁺+!, requires C 31 H 37 N 4O 2F 2: 535.2806).

[376]

[377] Example 28 : Preparation of

2-(2-(4-(bis(4-fluorophenyl)methyl)piperazin-l-yl)-2-oxoethylamino)-N-(3,4,5-tri methoxybenzyl)acetamide

[378]

[379] The title compound was prepared in the same manner as in Example 27, except that

N-((t-butyloxy)carbonyl)-N'-(3,4,5-trimethoxybenzyl)iminodiacetic acid monoamide was used instead of N-

((t-butyloxy)carbonyl)-N'-(l-methyl-3-phenylpropyl)iminodiacetic acid monoamide in Example 27 (white solid, yield: 63%). [380] mp 163-167⁰C;

[381] ¹H NMR (CDCl₃ 400MHz) δ 3.380~3.414(m, 4H), 3.791(s, 9H), 3.858~3.923(m,

2H), 4.116~4.118(m, 2H), 4.242~4.363(m, 4H), 4.485-4.516(m, 2H), 5.529(s, IH), 6.643~6.689(m, 2H), 7.199~7.242(m, 4H), 8.187~8.185(m, 4H);

[382] FABHRMS (m/z): 583.2732 (M⁺+l, requires C H N O F : 583.2654).

31 37 4 5 2

[383]

[384] Preparative Example 14 : Preparation of

4-(4-methylbenzylcarbamoyl)-3,3-dimethylbutanoic acid

[385] 2 mmol of 3,3-dimethylglutaric anhydride was added to 3 mL of dimethyl- formamide (DMF) to dissolve. Then, 2 mmol of 4-methylbenzylamine was added thereto, and stirred at room temperature for 20 hours. 20 mL of 10% HCl was put into the reaction solution, and extracted with 30 mL of ethylacetate (EtOAc). The organic layer was washed with 20 mL of 10% HCl, and then washed with 20 mL of a saturated NaCl solution twice. The organic layer was collected, dried over anhydrous MgSO , and filtered under reduced pressure. The organic solvent in the filtrate was removed under reduced pressure. The residue was recrystallized from a mixed solvent of EtOAc and methanol, so as to obtain the title compound (white solid, yield: 79%).

[386] mp 85-86⁰C;

[387] ¹U NMR (CDCl ) δ 12.09 (s, OH), 8.32 (t, J=5.8 Hz, NH), 7.10-7.15 (m, 4CH),

4.21 (d, J=6.0 Hz, CH₂), 2.28 (s, CH₂), 2.27 (s, CH₃), 2.19 (s, CH₂), 1.02 (s, 2CH₃);

[388] HR-FABMS Calcd for C H NO : (M⁺+l): 264.1600, Found: 264.1602.

[389]

[390] Preparative Example 15 : Preparation of

4-(4-methoxybenzylcarbamoyl)-3,3-dimethylbutanoic acid

[391] The title compound was prepared in the same manner as in Preparative Example 14, except that 4-methoxybenzylamine was used instead of 4-methylbenzylamine in Preparative Example 14 (white solid, yield: 85%).

[392] mp 102-103⁰C;

[393] ¹U NMR (CDCl ) δ 12.05 (s, OH), 8.32 (t, J=5.8 Hz, NH), 7.17 (d, J=8.0 Hz,

2CH), 6.87 (d, J=8.0 Hz, 2CH), 4.18 (d, J=5.6 Hz, CH ), 3.72 (s, OCH ), 2.28 (s, 2CH ), 1.04 (s, CH₃), 1.02 (s, CH₃);

[394] HR-FABMS Calcd for C H NO (M⁺+l): 280.1549, Found: 280.1542.

15 21 4

[395]

[396] Preparative Example 16 : Preparation of

4-(3,4,5-trimethoxybenzylcarbamoyl)-3,3-dimethylbutanoic acid

[397] The title compound was prepared in the same manner as in Preparative Example 14, except that 3,4,5-trimethoxybenzylamine was used instead of 4-methylbenzylamine in Preparative Example 14 (light yellow solid, yield: 54%). [398] mp 118-119⁰C;

[399] ¹H NMR (CDCl 3 ) δ 12.07 (s, OH), 8.33 (t, J=5.8 Hz, NH), 6.56 (d, J=8.0 Hz,

2CH), 4.21 (d, J=6.0 Hz, CH ), 3.74 (s, 2OCH ), 3.62 (s, OCH ), 2.29 (s, CH ), 2.21 (s, CH₂), 1.05 (s, 2CH₃); [400] HR-FABMS Calcd for C H NO : (M⁺+l): 340.1760, Found: 340.1757.

17 25 6

[401]

[402] Preparative Example 17 : Preparation of

4-(cyclohexylcarbamoyl)-3,3-dimethylbutanoic acid

[403] The title compound was prepared in the same manner as in Preparative Example 14, except that cyclohexylamine was used instead of 4-methylbenzylamine in Preparative

Example 14 (white solid, yield: 67%). [404] mp 77-78⁰C;

[405] ¹U NMR (CDCl ) δ 12.07 (s, OH), 7.76 (d, J=8.0 Hz, NH), 3.53 (s, CH), 2.25 (s,

CH₂), 2.09 (s, CH₂), 1.52-1.72 (m, 2CH₂), 1.04-1.30 (m, 3CH₂), 1.05 (s, CH₃), 1.01 (s,

CH₃); [406] HR-FABMS Calcd for C 13 H 23 NO 3 : (M⁺+l): 242.1756, Found: 242.1752.

[407]

[408] Example 29 : Preparation of N-

(4-methylbenzyl)-5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-3,3-dimet hyl-5-oxopentanamide

[409]

[410] 0.5 mmol of 4-(4-methylbenzylcarbamoyl)-3,3-dimethylbutanoic acid prepared in

Preparative Example 14, 0.55 mmol of l-(4-chlorobenzhydryl)piperazine, and ben- zotriazol-1-yloxytripyrrolidino phosphonium hexafluorophosphate (PyBOP) were added to 3 mL of dimethylformamide (DMF) to dissolve. Then, 1.0 mmol of N,N-diisopropylethylamine (DIEA) was added thereto, and stirred at room temperature for 16 hours. 20 mL of 10% HCl was put into the reaction solution, and extracted with 30 mL of EtOAc. The organic layer was washed with 20 mL of 10% HCl, and then washed with 20 mL of a saturated NaHCO 3 solution twice and with 20 mL of a saturated NaCl solution twice. The organic layer was collected, dried over anhydrous

MgSO , and filtered under reduced pressure. The organic solvent in the filtrate was

4 removed under reduced pressure. The residue was purified by column chromatography with a mixed solvent of EtOAc and methanol (20:1), so as to obtain the title compound (light yellow solid, yield: 70%). [411] mp 63-64°C;

[412] ¹U NMR (CDCl ) δ 8.05 (t, J=5.8 Hz, NH), 7.35 (d, J=8.0 Hz, 4CH), 7.30 (d, J=7.2

Hz, 3CH), 7.26 (t, J=8.4 Hz, 2CH), 7.18 (d, J=8.0 Hz, 2CH), 7.10 (d, J=8.0 Hz, 2CH), 4.38 (d, J=5.6 Hz, CH₂), 4.20 (s, CH), 3.56-3.66 (m, CH₂), 3.51 (t, J=5.0 Hz, CH₂),

2.34-2.37 (m, 2CH ), 2.33 (s, CH ), 2.32 (s, 2CH ), 1.02 (s, 2CH ); [413] HR-FABMS Calcd for C 32 H 38 ClN 3 O 2 : (M⁺+l): 532.2731, Found: 532.2719.

[414]

[415] Example 30 : Preparation of N-

(4-methoxybenzyl)-5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-3,3-dime thyl-5-oxopentanamide

[416]

[417] The title compound was prepared in the same manner as in Example 29, except that

4-(4-methoxybenzylcarbamoyl)-3,3-dimethylbutanoic acid prepared in Preparative Example 15 was used instead of 4-(4-methylbenzylcarbamoyl)-3,3-dimethylbutanoic acid in Example 29 (white solid, yield: 63%).

[418] mp 64-65⁰C;

[419] ¹U NMR (CDCl ) δ 8.06 (t, J=5.6 Hz, NH), 7.35 (d, J=6.4 Hz, 4CH), 7.29 (t, J=8.1

Hz, 2CH), 7.26 (d, J=8.8 Hz, 2CH), 7.21 (t, J=8.8 Hz, CH), 6.83 (d, J=8.8 Hz, 4CH), 4.35 (d, J=6.0 Hz, CTT), 4.20 (s, CH), 3.79 (s, OCH₃), 3.56-3.65 (m, CH₂), 3.51 (t, J=5.0 Hz, CH₂), 2.33-2.39 (m, 2CH₂), 2.32 (s, CH₂), 2.30 (s, CH₂), 1.05 (s, 2CH₃);

[420] HR-FABMS Calcd for C 32 H 38 ClN 3 O 3 : ( ^VM⁺+l) ': 548.2680, Found: 548.2684.

[421]

[422] Example 31 : Preparation of N-

(3,4,5-trimethoxybenzyl)-5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-3,

3-dimethyl-5-oxopentanamide

[423]

[424] The title compound was prepared in the same manner as in Example 29, except that

4-(3,4,5-trimethoxybenzylcarbamoyl)-3,3-dimethylbutanoic acid prepared in Preparative Example 16 was used instead of 4-(4-methylbenzylcarbamoyl)-3,3-dimethylbutanoic acid in Example 29 (white solid, yield: 46%).

[425] mp 67-68⁰C;

[426] ¹U NMR (CDCl ) δ 8.37 (t, J=5.8 Hz, NH), 7.35 (d, J=8.4 Hz, 4CH), 7.29 (t, J=7.6

Hz, 2CH), 7.26 (d, J=8.8 Hz, 2CH), 7.21 (t, J=7.2 Hz, CH), 6.83 (d, J=8.8 Hz, 2CH), 4.37 (d, J=6.0 Hz, CH₂), 4.20 (s, CH), 3.82 (s, 3OCH₃), 3.56-3.67 (m, CH₂), 3.53 (t, J=5.0 Hz, CH ), 2.34-2.41 (m, 2CH ), 2.34 (s, CH ), 2.30 (s, CH ), 1.06 (s, 2CH );

[427] HR-FABMS Calcd for C 34 H 42 ClN 3 O 5 : (M⁺+l): 608.2891, Found: 608.2861.

[428]

[429] Example 32 : Preparation of N- cyclohexyl-5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-3,3-dimethyl-5-o xopentanamide

[430]

[431] The title compound was prepared in the same manner as in Example 29, except that

4-(4-cyclohexylcarbamoyl)-3,3-dimethylbutanoic acid prepared in Preparative Example 17 was used instead of 4-(4-methylbenzylcarbamoyl)-3,3-dimethylbutanoic acid in Example 29 (white solid, yield: 9%).

[432] mp 77-78⁰C;

[433] ¹U NMR (CDCl ) δ 7.65 (d, J=8.4 Hz, NH), 7.35 (d, J=8.4 Hz, 2CH), 7.29 (t, J

=7.4 Hz, 2CH), 7.25 (t, J=8.0 Hz, CH), 7.22 (d, J=6.8 Hz, 2CH), 7.21 (d, J=6.4 Hz, 2CH), 4.21 (s, CH), 3.76 (s, CH), 3.60-3.70 (m, CTT), 3.55 (t, J=5.0 Hz, CH₂), 2.30-2.43 (m, 2CH ), 2.24 (s, CH ), 2.17 (s, CH ), 1.86 (d, /=12.4 Hz, CH ), 1.67 (d, / = 12.0 Hz, CH₂), 1.29-1.40 (m, CH₂), 1.10-1.28 (m, 2CH₂), 1.05 (s, CH₃), 1.01 (s, CH₃);

[434] HR-FABMS Calcd for C 30 H 40 ClN 3 O 2 : (M⁺+l): 510.2887, Found: 510.2874.

[435]

[436] Experimental Example 1 : Inhibitory effect of compound of the invention on bradykinin activity

[437] In order to confirm the inhibitory effect of the compound of the invention on bradykinin activity, the following experiment was performed using a guinea-pig ileum.

[438] Male hartley guinea-pigs with body weight of 275 to 500 g were used as laboratory animals. The animals were fasted overnight before experiments, and beheaded. 40 cm of specimen was taken by cutting from 2 cm above the ileocecal junction, and immersed in a Tyrode's solution (mM) [NaCl 136.9, KCl 2.7, CaCl 1.8, MgCl 1.15, NaH PO 0.4, NaHCO 11.9, glucose 5.6] at 37⁰C supplied with a gas mixture of 95% 0 / 5% CO . Among them, the distal 10 cm of ileum was used. The ileum muscle was cut into 1.5 to 2 cm, and then placed in 50 ml of organ bath containing the Tyrode's solution at 37⁰C. The gas mixture of 95% O / 5% CO was injected into the Tyrode's solution. After stabilizing for 1 hour, 1 μM of each compound prepared in Example 1 to 32 was added thereto. After 15 minutes, bradykinin (0.1 μM) was added thereto. The contractile response to bradykinin was observed twice to three times every 20 minutes, and thus the sensitivity and reproducibility of the ileum contractile response were measured using a Grass model 76E polygraph. The ileum, in which the reproducibility was confirmed, were used.

[439] To prevent degradation of bradykinin, neuronal activation, and prostaglandin production, 1 μM of each of captopril, atropine, dithiothreitol, and indomethacin was added to the Tyrode's solution to use. Moreover, to prevent histaminergic responses, 1 μM of dibenamine, which is an irreversible histamine H blocker, was also added thereto. HOE 140 was used as a control.

[440] The results are shown in Table 1. [441] Table 1

[442] [443] As shown in Table 1, it can be seen that the compound according to the present invention has excellent inhibitory effect on bradykinin activity.

[444] [445] Experimental Example 2 : Concentration-dependent inhibitory effect of compound of the invention on bradykinin activity

[446] In order to confirm the inhibitory effect of the compound of the invention on bradykinin activity according to changes in its concentration, the same method as Experimental Example 1 was performed by varying the concentration of bradykinin (1 nM to 10 μM).

[447] Four compounds (Examples 6, 15, 16, 20) were screened according to their pharmacological activities and structures. In order to confirm the maximal response of the ileum, 80 mM KCl was added at the end of each curve. After washing and stabilizing, the bradykinin activities were measured varying the concentration of the compounds of Examples 6, 15, 16, and 20 (10 nM to 1 μM). The same experiment was performed using HOE 140 as a control, by varying its concentration (3 nM to 1 μM).

[448] The results are shown in Table 2. [449] Table 2

[450] [451] As shown in Table 2, it can be seen that even a small amount of the compound according to the invention can effectively inhibits bradykinin activity.

[452] [453] Experimental Example 3 : Analgesic effect of compound of the invention on chronic neuropathic pain

[454] In order to confirm the analgesic effect of the compound of the invention on chronic neuropathic pain, experiments were performed as follows. [455] 1. Induction of chronic neuropathic pain - Preparation of caudal nerve injury model [456] 8 week-Sprague-Dawley rats were divided into experimental groups of four rats each. The rats were housed with a 12 hr light/dark cycle (light on at 7 am) and free access to food and water at 22 to 25⁰C. [457] In order to induce chronic neuropathic pain, the rats were anesthetized with 0.5 to

2% enflurane, and followed by transection of the inferior/superior caudal trunks between the Sl and S2 spinal nerves that innervated the rat tails (Fig. 1). To prevent the possible rejoining of the proximal and distal ends of the severed trunk, about 1 mm piece of the trunk was removed from the proximal end.

[458] This surgery eliminated the Sl spinal nerve innervation of the tail, so as to give a model having chronic neuropathic pain.

[459] 2. Measurement of analgesic effect on chronic neuropathic pain

[460] In order to test the chronic neuropathic pain in the tail of the laboratory animal

(mechanical, cold, and warm allodynia tests), behavioral tests were performed on day 1 before surgery, and on days 1, 7, 14 after surgery. To apply mechanical, cold, and warm stimuli to tails, the laboratory animals were restrained in a transparent plastic tube (4.5x12, 5.5x5, 6.5x18 cm; diameterxlength). The laboratory animals were adapted to the test environment for 1 hour before behavioral tests.

[461] At day 14 after inducing nerve injury, when allodynia of the laboratory animal model prepared in 1 reaches to a maximum, 60 mg/kg of each compound prepared in Examples 6 and 26 were intraperitoneally administered. After 1, 3, and 5 hours, the behavioral tests were performed to measure anti-allodynic effect of the compound of the present invention. At this time, a vehicle control was used as a control.

[462] For mechanical allodynia test, the withdrawal threshold was determined using an up-down method. A series of eight von Frey filaments (0.4, 0.6, 1.0, 2.0, 4.0, 6.0, 8.0, 15.0 g, Stoelting, Wood Dale, IL, USA) were used. The tail flick to a von Frey application was regarded as a withdrawal response. The first stimulus used was the 2.0 g filament, and when a withdrawal response was obtained, the next weaker filament was used. When no response was obtained, the next stronger filament was administered. Interpolation of the 50% threshold was carried out using the Dixon method.

[463] For cold and warm allodynia test, a withdrawal latency was evaluated. The tail of the laboratory animal was immersed in cold water (4⁰C) and warm water (4O⁰C) baths, and then withdrawal latency was measured within a cut-off time of 15 seconds. The test for each animal was repeated five times at 5-min intervals to obtain the mean latency of tail withdrawals. Shorter latency was interpreted as more severe cold and warm allodynia.

[464] The inhibitory effects of the compounds of Examples 6 and 26 on chronic neuropathic pain are shown in each Fig. 2 and Fig. 3 (A: mechanical allodynia, B: cold allodynia, and C: warm allodynia). In Fig. 2 and Fig. 3, a perpendicular dotted line represents the time of injecting the compound, and n represents the number of rats used in the experiment.

[465] As shown in Fig. 2 and Fig. 3, higher analgesic effect on chronic neuropathic pain was observed in the group treated with the compound of the invention, as compared to the experimental group treated with the vehicle control. A statistically significant analgesic effect on the stimulus inducing cold allodynia was occurred at 1 hour after injecting the compound of the invention (P<0.05). Further, a statistically significant analgesic effect on the stimulus inducing warm allodynia was occurred at 3 hours after injecting the compound of the invention (P<0.05).

[466]

[467] Experimental Example 4 : Inhibitory effect of compound of the invention on inflammatory pain

[468] In order to confirm the inhibitory effect of the compound of the invention on inflammatory pain, the following experiments were performed using formalin, which is a material inducing inflammation.

[469] 5% Formalin 50 D and the compounds prepared in Examples 3, 6, 14, 20, 26, and 27

(each 60 D/D) were injected subcutaneously into the one hindpaw of the rat (6 D/D of the compound prepared in Example 6 was injected subcutaneously into the laboratory animal). As a control, only 5% Formalin 50 D was injected subcutaneously into the one hindpaw of the rat. Then, the rats were placed in an observation chamber (width, length 16 cmxl4 cm). The time spent in licking and biting the injected paw was recorded every 5 minutes over a 60-min period. Phase I was defined as the first 10 min after formalin injection, phase II was defined as the following 50 min, and the results was analyzed.

[470] The inhibitory effects of the compounds of Examples 3, 6, 14, 20, 26, and 27 of the invention on inflammatory pain are shown in Figs. 4 to 9, respectively.

[471] As shown in Fig. 4, the analgesic effect on inflammatory pain in the group treated with the compound of the invention was observed in the phase II (a period of 10 min to 60 min after injection), and no analgesic effect was observed in the phase I (until 10 min after injection), as compared to the control. In Figs. 5 to 9, excellent analgesic effect on inflammatory pain in the group treated with the compound of the invention was also observed in the phase II, as compared to the control.

[472]

[473] Hereinbelow, Formulation Examples for the composition of the present invention will be illustrated.

[474] Formulation Example 1 : Preparation of liquid injectable formulation

[475] An injectable liquid formulation containing 10 mg of the active ingredient was prepared in the following manner.

[476] 1 g of the compound of the formula 1, 0.6 g of sodium chloride, and 0.1 g of ascorbic acid were dissolved in distilled water to be 100 ml. The solution was put into a bottle, and heated to sterilize at 2O⁰C for 30 minutes. [477] The composition of the injectable liquid formulation is as follows.

[478] Compound of formula 1 1 g

[479] Sodium chloride 0.6 g

[480] Ascorbic acid 0.1 g

[481] Distilled water q.s.

[482]

[483] Formulation Example 2 : Preparation of syrup formulation

[484] A syrup formulation containing the compound of the formula 1 as an active ingredient (2%, weight/volume) was prepared in the following manner. [485] The compound of the formula 1, saccharin, and sugar were dissolved in 80 g of warm water. The solution was cooled, and a solution containing glycerin, saccharin, flavor, ethanol, sorbic acid, and distilled water was added thereto. Water was added to the mixture to be 100 ml.

[486] The composition of the syrup formulation is as follows.

[487] Compound of formula 1 2 g

[488] Saccharin 0.8 g

[489] Sugar 25.4 g

[490] Glycerin 8.0 g

[491] Flavor 0.04 g

[492] Ethanol 4.0 g

[493] Sorbic acid 0.4 g

[494] Distilled water q.s.

[495]

[496] Formulation Example 3 : Preparation of tablet formulation

[497] A tablet formulation containing 15 mg of the active ingredient was prepared in the following manner. [498] 250 g of the compound of the formula 1 was mixed with 175.9 g of lactose, 180 g of starch, and 32 g of colloidal silicic acid. 10% Gelatin solution was added to the mixture, and then pulverized to pass through a 14-mesh sieve. The mixture was dried.

Then, 160 g of starch, 50g of talc, and 5 g of magnesium stearate were added thereto to prepare a tablet.

[499] The composition of the tablet formulation is as follows.

[500] Compound of formula 1 250 g

[501] Lactose 175.9 g

[502] Starch 180 g

[503] Colloidal silicic acid 32 g

[504] 10% Gelatin solution

[505] Starch 160 g [506] Talc 50 g

[507] Magnesium stearate 5 g

Industrial Applicability

[508] The non-peptide compound of the present invention has excellent inhibitory effect on bradykinin activity, and on chronic neuropathic pain and inflammatory pain, thereby being used for preventing or treating a disease caused by bradykinin, in particular, pain.

Claims

Claims[1] A non-pep tide compound represented by the following formula 1 or a pharmaceutically acceptable salt thereof: [Formula 1] whereinRl and R2 are each independently hydrogen; C ~C alkyl; C ~C alkoxy; C ~C cycloalkyl; C ~C aryl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; C ~C heteroaryl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C 1 ~C4 alkyl, C 1 ~C4 alkoxy, and halogen; C ~C alkyl substituted with C ~C aryl which is substituted or un-° 1 4 J 6 20 substituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; or C ~C alkyl substituted with C ~C4 ^ 1 4 ° 1 4 ^ 5 20 heteroaryl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen, X is C ~C alkylenyl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; C ~C alkenylenyl which is substituted or unsubstituted with at least2 6 one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; -CH YCH -; or -CH CH Y-,Y is O, S, or NR3,R3 is hydrogen, C 1 ~C4 alkyl, C1 ~C4 alkoxy, halogen or -COO-C 1 ~C4 alkyl,Z and Z' are each independently hydrogen, hydroxy, or halogen, and n is an integer of 1 to 3(provided that if n is 2, X is not -CH YCH - (wherein Y is NR3, and R3 is hydrogen)).[2] The non-pep tide compound or the pharmaceutically acceptable salt thereof according to claim 1, whereinRl and R2 are each independently hydrogen; C ~C alkyl; C ~C cycloalkyl; or C ~C alkyl substituted with C ~C aryl which is substituted or unsubstituted1 4 6 20 with at least one substituent selected from the group consisting of C 1 ~C4 alkyl, C 1~C alkoxy, and halogen, X is C ~C alkylenyl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; C ~C alkenylenyl which is substituted or unsubstituted with at least2 6 one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; or -CH YCH -,2 2Y is NR3,R3 is hydrogen or -COO-C ~C alkyl,1 4Z and Z' are each independently hydrogen or halogen, and n is an integer of 2 or 3(provided that if n is 2, X is not -CH YCH - (wherein Y is NR3, and R3 is hydrogen)). [3] The non-peptide compound or the pharmaceutically acceptable salt thereof according to claim 1, whereinRl and R2 are each independently hydrogen, methyl, cyclohexyl,4-methylbenzyl, 4-methoxybenzyl, 3,4,5-trimethoxybenzyl, or1 -methyl-3-phenylpropyl,X is ethylenyl, 2-methylpropylenyl, 2,2-dimethylpropylenyl, ethenylenyl, -CHNHCH -, or -CH NCOO(t-butyl)CH -,Z and Z' are each independently hydrogen, F, or Cl, and n is an integer of 2 or 3(provided that if n is 2, X is not -CH NHCH -). [4] The non-peptide compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is selected from the group consisting of1)5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-N-(4-methylbenzyl)-3-met hyl- 5 -oxopentanamide ;2)5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-N-(4-methoxybenzyl)-3-m ethyl- 5 -oxopentanamide ;3)5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-N-(3,4,5-trimethoxybenzy l)-3-methyl-5-oxopentanamide;4)5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-N-cyclohexyl-3-methyl-5- oxopentanamide ;5) 5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-N,N,3-trimethyl-5-oxopen tanamide;6)5-(4-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan-l-yl)-N-(4-methylbenzyl)-3-methyl-5-oxopentanamide;7) 5-(4-((4-chlorophenyl)(phen yl)methyl)- 1 ,4-diazepan- 1 -yl)-N-(4-methoxybenzyl)-3-methyl-5-oxopentanamid e;8)5-(4-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan-l-yl)-N-(3,4,5-trimethoxyb enzyl)- 3 -methyl- 5 -oxopentanamide ;9)5 - (4- ( (4-chlorophenyl) (phenyl)methyl) - 1 ,4-diazepan- 1 -yl) -N-cyclohexyl- 3 -meth yl- 5 -oxopentanamide ;10) t-butyl (dimethylcarbamoyl)methyl2-(4-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan-l-yl)-2-oxoethylcarbamate;H)2-(2-(4-((4-cmorophenyl)(phenyl)methyl)-l,4-diazepan-l-yl)-2-oxoethylamino)- N,N-dimethylacetamide ;12) N- (4-methylbenzyl)-4-oxo-4-((4-chlorobenzhydryl)piperazin-l-yl)butanamide;13) N- (4-methoxybenzyl)-4-oxo-4-((4-chlorobenzhydryl)piperazin-l-yl)butanamide;14) N-(3,4,5-trimethoxybenzyl)-4-oxo-4-((4-chlorobenzhydryl)piperazin-l-yl) butanamide;15) N-(cyclohexyl)-4-oxo-4-((4-chlorobenzhydryl)piperazin-l-yl)butanamide;16) N-(4-methylbenzy l)-4-oxo-4-((4-chlorobenzhydryl)- 1 ,4-diazepan- 1 -yl)butanamide;17) N-(4-methoxybenzyl)-4-oxo-4-((4-chlorobenzhydryl)-l,4-diazepan-l-yl)butanamid e;18) N-(3,4,5-trimethoxybenzyl)-4-oxo-4-((4-chlorobenzhydryl)-l,4-diazepan-l-yl)buta namide;19) N-(cyclohexyl)-4-oxo-4-((4-chlorobenzhydryl)- 1 ,4-diazepan- 1 -yl)butanamide;20) N-(3,4,5-trimethoxybenzyl)-N'-l-((4-chlorobenzhydryl)-l,4-diazepan-l-yl)iminodi acetic acid diamide; 21)(Z)-N-(4-methylbenzyl)-4-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-4-oxo-but-2-enamide;22)(Z)-N-(4-methoxybenzyl)-4-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-4-oxo-but-2-enamide;23)(Z)-4-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-4-oxo-N-(4-phenylbut an-2-yl)-but-2-enamide;24)(Z)-N-(4-methylbenzyl)-4-(4-(bis(4-fluorophenyl)methyl)piperazin-l-yl)-4-oxo- but-2-enamide;25) (Z)-N-(4 - methoxybenzyl)-4-(4-(bis(4-fluorophenyl)methyl)piperazin-l-yl)-4-oxo-but-2-en amide;26)(Z)-4-(4-(bis(4-fluorophenyl)methyl)piperazin-l-yl)-4-oxo-N-(4-phenylbutan-2- yl)-but-2-enamide;27)2-(2-(4-(bis(4-fluorophenyl)methyl)piperazin-l-yl)-2-oxoethylamino)-N-(4-phen ylbutan-2-yl)acetamide;28)2-(2-(4-(bis(4-fluorophenyl)methyl)piperazin-l-yl)-2-oxoethylamino)-N-(3,4,5-tr imethoxybenzyl) acetamide ;29) N-(4-methylbenzyl)-5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-3,3-dim ethyl- 5 -oxopentanamide ;30) N-(4-methoxybenzyl)-5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-3,3-di methyl- 5 -oxopentanamide ;31) N-(3,4,5-trimethoxybenzyl)-5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)- 3,3-dimethyl-5-oxopentanamide; and32) N- cyclohexyl-5-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-l-yl)-3,3-dimethyl-5 -oxopentanamide.[5] A method for preparing the non-peptide compound or the pharmaceutically acceptable salt thereof according to claim 1, comprising the steps of 1) preparing a compound of the following formula 4 by condensation reaction of a compound of the following formula 2 and an amine compound of the following formula 3; and2) preparing the compound of the following formula 1 by condensation reaction of the compound of the following formula 4 and a compound of the following formula 5,

[Formula 1]

[Formula 2]

[Formula 3] R1R2NH

[Formula 4]

O O

R1 -

^"N X OH R2

[Formula 5]

wherein

Rl and R2 are each independently hydrogen; C ~C alkyl; C ~C alkoxy; C ~C

1 4 1 4 3 10 cycloalkyl; C ~C aryl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; C ~C heteroaryl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; C ~C alkyl substituted with C ~C aryl which is substituted or un-

^& 1 4 ^J 6 20 ^J substituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; or C ~C alkyl substituted with C ~C

4 1 4 1 4 5 20 heteroaryl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen, X is C ~C alkylenyl which is substituted or unsubstituted with at least one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; C ~C alkenylenyl which is substituted or unsubstituted with at least

2 6 one substituent selected from the group consisting of C ~C alkyl, C ~C alkoxy, and halogen; -CH YCH -; or -CH CH Y-,

2 2 2 2

Y is O, S, or NR3,

R3 is hydrogen, C ~C alkyl, C ~C alkoxy, halogen, or -COO-C ~C alkyl,

1 4 1 4 1 4

Z and Z' are each independently hydrogen, hydroxy or halogen, and n is an integer of 1 to 3

(provided that if n is 2, X is not -CH 2 YCH 2 - (wherein Y is NR3, and R3 is hydrogen)).

[6] The method for preparing the non-peptide compound or the pharmaceutically acceptable salt thereof of claim 1 according to claim 5, wherein the compound of the formula 2 is 3-methylglutaric anhydride or succinic anhydride.

[7] The method for preparing the non-peptide compound or the pharmaceutically acceptable salt thereof of claim 1 according to claim 5, wherein the amine compound of the formula 3 is 4-methylbenzylamine, 4-methoxybenzylamine, 3,4,5-trimethoxybenzylamine, cyclohexylamine, dimethylamine, or 1 -methyl-3-phenylpropylamine.

[8] The method for preparing the non-peptide compound or the pharmaceutically acceptable salt thereof of claim 1 according to claim 5, wherein the compound of the formula 5 is l-(4-chlorobenzhydryl)piperazine or l-((4-chlorophenyl)(phenyl)methyl)-l,4-diazepan.

[9] The method for preparing the non-peptide compound or the pharmaceutically acceptable salt thereof of claim 1 according to claim 5, wherein the reaction solvent used in the step 1) and 2) of the condensation reaction is one selected from the group consisting of ethyl ether, THF (tetrahydrofuran), dichloromethane, chloroform, DMSO (dimethyl sulfonyloxide), and DMF(Dimethylformamide) .

[10] A pharmaceutical composition for preventing or treating a disease caused by bradykinin, comprising the non-peptide compound or the pharmaceutically acceptable salt thereof of claim 1.

[11] The pharmaceutical composition according to claim 10, wherein the disease caused by bradykinin is one selected from the group consisting of asthma, allergic rhinitis, arthritis, rheumatic arthritis, septic shock, hemorrhagic shock, hypersensitivity, cerebral edema, angioneurotic edema, acute pancreatitis, postgastrectomy dumping syndrome, carcinoid syndrome, migraine, and pain.

[12] The pharmaceutical composition according to claim 11, wherein the disease caused by bradykinin is pain.

[13] The pharmaceutical composition according to claim 12, wherein the pain is chronic neuropathic pain or inflammatory pain.