WO2008013334A1

WO2008013334A1 - Deformylase inhibitor, process for the preparation thereof, and composition comprising the same

Info

Publication number: WO2008013334A1
Application number: PCT/KR2006/003639
Authority: WO
Inventors: Bong-Jin Lee; Seung-Kyu Lee; Kwang-Hyun Choi; Jong-Sun Lee
Original assignee: Promeditech Inc.
Priority date: 2006-07-28
Filing date: 2006-09-13
Publication date: 2008-01-31
Also published as: KR100753796B1

Abstract

Provided are a novel compound useful for a deformylase inhibitor with excellent antibacterial activity or its pharmaceutically acceptable salt, a process for preparing the same, and an antibacterial composition including the same as an active ingredient. The deformylase inhibitor has a broad spectrum against bacteria, including bacteria with resistance to existing antibacterial agents.

Description

DEFORMYLASE INHIBITOR, PROCESS FOR THE

PREPARATION THEREOF, AND COMPOSITION

COMPRISING THE SAME

Technical Field

[1] The present invention relates to a novel compound or its pharmaceutically acceptable salt useful for a deformylase inhibitor with excellent antibacterial activity, a process for preparing the same, and an antibacterial composition comprising the same as an active ingredient. Background Art

[2] Deformylase is metallopeptidase found in prokaryotes such as bacteria. In prokaryotes, protein synthesis is initiated with N-formyl methionine (fMet). The formyl group is removed by deformylase before a protein reaches its mature form. Since such deformylation is a prerequisite for protein maturation, it is known that deformylase is essential for bacterial growth (Chang et al., J. Bacteriol. Ill: 4071-4072(1989); Meinnel T, Blanquet S, /. Bacteriol. 176(23): 7387-90(1994); and Mazel D et. al., EMBO J. 13(4): 914-23(1994)). However, fMet is not a requisite for initiation of eukaryotic protein synthesis, and thus, deformylase inhibitors can be effectively used as antibacterial agents with broad-spectrum.

[3] Deformylase inhibitors are disclosed in WO 02/102791 (pyrrolidine bicyclic derivatives), WO 02/102790 (N-formyl hydroxyl amine derivatives), WO 01/44179 (succinate derivatives), WO 01/44178 (urea derivatives), and WO 01/85170 (peptide derivatives).

[4] However, development of novel deformylase inhibitors having a broad spectrum against bacteria, including bacteria with resistance to existing antibacterial agents, is still required.

Disclosure of Invention Technical Problem

[5] The present invention provides a novel compound useful for a deformylase inhibitor having an amide bond, i.e., a non-peptide bond, and a process for preparing the same. The present invention also provides an antibacterial composition comprising the compound as an active ingredient. Technical Solution

[6] In accordance with an aspect of the present invention, there is provided a compound of formula (I) or its pharmaceutically acceptable salt: [7]

[8] wherein, R is a straight or branched C ~C alkyl group,

2 ^{X 6} [9] R is hydrogen or a straight or branched C ~C alkyl group, and

3 ' ⁶ [10] R is a straight or branched C ~C alkyl group, a pyridinyl group, a phenyl group, a

1 6 quinolinyl group, or a isoquinolinyl group (wherein, the pyridinyl group, the phenyl group, the quinolinyl group, or the isoquinolinyl group may be substituted with 1 to 3 substituent(s) selected from the group consisting of C -C alkyl, C -C alkoxy,

1 4 1 4 halogen, and cyano).

[H] In another aspect of the present invention, there is provided a process for preparing a compound of formula (T) or its pharmaceutically acceptable salt, which comprises performing a deprotection reaction of a compound of formula (III):

[12]

[13] [14] wherein, R¹, R², and R³ are the same as defined above and Y is a hydroxy- protecting group.

[15] In still another aspect of the present invention, there is provided an antibacterial composition comprising a therapeutically effective amount of a compound of formula (I) or its pharmaceutically acceptable salt and a pharmaceutically acceptable carrier. Mode for the Invention

[16] In one aspect of the present invention, there is provided a compound of formula (I) or its pharmaceutically acceptable salt: [17]

[18] wherein, R is a straight or branched C ~C alkyl group,

1 6 [19] R^ is hydrogen or a straight or branched C ~C alkyl group, and

3 ' ⁵ [20] R is a straight or branched C ~C alkyl group, a pyridinyl group, a phenyl group, a

1 6 quinolinyl group, or a isoquinolinyl group (wherein, the pyridinyl group, the phenyl group, the quinolinyl group, or the isoquinolinyl group may be substituted with 1 to 3 substituent(s) selected from the group consisting of C ~C_^ alkyl, C 1-C 4_^ alkoxy, halogen, and cyano).

[21] [22] Among the compounds of formula (I) or pharmaceutically acceptable salts thereof,

^3 preferred are those wherein R is a radical of formula (TIa), (lib), or (He):

[23]

[24] [25] wherein, R , R , R , and R , independently each other, hydrogen, a C -C alkyl

1 4 group, a C ~C alkoxy group, a halogen group, or a cyano group, and * is a binding

1 4 site. [26] [27] More preferred compounds of formula (I) or pharmaceutically acceptable salts thereof are: [28] pyridin-2-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [29] 6-methyl-pyridin-2-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [30] 4-chloro-pyridin-2-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [31] quinolin-2-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [32] isoquinolin-l-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [33] 6-fluoro-pyridin-2-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [34] 3-methyl-pyridin-2-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [35] pyridin-3-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [36] pyridin-4-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [37] 3-fluoro-pyridin-2-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [38] 5-methyl-pyridin-2-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [39] 4-methoxy-pyridin-2-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]

- 1 -isopropyl-2-oxo-heptyl } -amide; [40] 5-fluoro-pyridin-2-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [41] S-methyl-pyridin-S-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [42] 2-methoxy-pyridin-3-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]

- 1 -isopropyl-2-oxo-heptyl } -amide; [43] 2-fluoro-pyridin-3-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [44] ό-fluoro-pyridin-S-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [45] S-fluoro-pyridin-S-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [46] 2-methyl-pyridin-3-carboxylic acid {(lS,3R)-3-[(formyl-hydτoxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [47] 4-methyl-pyridin-3-carboxylic acid {(lS,3R)-3-[(formyl-hydτoxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [48] 2-methyl-pyridin-4-carboxylic acid {(lS,3R)-3-[(formyl-hydτoxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [49] 3-methyl-pyridin-4-carboxylic acid {(lS,3R)-3-[(formyl-hydτoxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [50] 2-cyano-pyridin-4-carboxylic acid {(lS,3R)-3-[(foπnyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [51] 2-methoxy-pyridin-4-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]

- 1 -isopropyl-2-oxo-heptyl } -amide; [52] 2-fluoro-pyridin-4-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [53] 3-fluoro-pyridin-4-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [54] phenyl-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [55] 3-methyl-phenyl-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [56] 2-fluoro-phenyl-carboxylic acid {(lS,3R)-3-[(foimyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [57] 4-methyl-phenyl-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [58] 3-methoxy-phenyl-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; [59] 3-fluoro-phenyl-carboxylic acid {(lS,3R)-3-[(foimyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide ; and [60] isopropyl-carboxylic acid {(IS, 3R)-3- [(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide. [61] [62] A compound of the present invention may be in the form of a pharmaceutically acceptable salt, hydrate(s), or solvate(s). The pharmaceutically acceptable salts which may be applied to a compound of the present invention include, but are not limited to, hydrochloride, hydrobromide, sulfate, methylsulfonate, p-tolunesulfonate, phosphate, acetate, citrate, succinate, lactate, tartarate, fumarate, malate, a sodium salt, a potassium salt, a magnesium salt, and a calcium salt.

[63] A compound of the present invention may also be in the form of racemates or optical isomers due to presence of chiral centers. Therefore, a compound of the present invention includes both racemates and optical isomers.

[64] In accordance with another aspect of the present invention, there is provided a process for preparing a compound of formula (T) or its pharmaceutically acceptable salt. In other words, there is provided a process for preparing a compound of formula (I) or its pharmaceutically acceptable salt, which comprises performing a deprotection reaction of a compound of formula (III):

[65]

[66] [67] wherein, R , R , and R are the same as defined above and Y is a hydroxy- protecting group.

[68] The hydroxy-protecting group (Y) may be a conventionally used protecting group, such as benzyl, benzyloxymethyl, and t-butoxymethyl. [69] The deprotection of a hydroxy-protecting group may be carried out by conventional deprotecting methods in the field of organic chemistry. For example, the hydroxy- protecting group may be removed by hydrogenating with Pd/C (palladium on activated charcoal) or adding trifluoroacetic acid. The deprotection reaction may be carried out at room temperature in an organic solvent such as C -C alcohol (e.g., methanol, ethanol), ethyl acetate, tetrahydrofuran, hexane, and toluene.

[70] The compound of formula (III) may be obtained by a process which comprises reacting a compound of formula (IV) or its salt with HOC(O)R³ (R³ is the same as defined above):

[71]

[72] wherein, R and R are the same as defined above and Y is a hydroxy-protecting group. [73] The salts of a compound of formula (IV) include, but are not limited to, hydrochloride and trifluoroacetate. Reacting the compound of formula (TV) or its salt with HOC(O)R may be carried out with a carboxylic acid-activating agent, e.g., N,N'-dicyclohexylcarbodiimide, N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide hydrochloride, in the presence of 1-hydroxybenzotriazole. And, the reaction may be carried out in the presence of a base such as N,N-diisopropylethylamine, in an organic solvent such as dichloromethane, and at 0 ~ 25 ⁰C.

[74] The compound of formula (IV) or its salt may be obtained by a process which comprises deprotecting a compound of formula (V): [75]

[76] wherein, R , R , and Y are the same as defined above and Z is an amino protecting group. [77] The amino protecting group may be conventionally used protecting groups, such as t-butoxycarbonyl, benzyloxycarbonyl, and 9-fluorenylmethyloxycarbonyl. [78] The deprotection of an amino protecting group may be carried out by conventional deprotecting methods in the field of organic chemistry. For example, the amino- protecting group may be removed by adding hydrochloric acid or trifluoroacetic acid, Pd/C and hydrogen, piperazine, and etc. The deprotection reaction may be carried out at room temperature in an organic solvent such as dichloromethane, C -C alcohol, tetrahydrofuran, and toluene.

[79] The compound of formula (V) may be obtained by a process which comprises reacting a compound of formula (VI) with formic acid and acetic anhydride: [80]

[81] wherein, R , R , Y, and Z are the same as defined above. [82] Reacting the compound formula (VI) with formic acid and acetic anhydride may be carried out at 0 ~ 25 ⁰C in an organic solvent such as ethyl acetate, and etc.

[83] The compound of formula (VI) may be obtained by a process which comprises reacting a compound of formula (VII) with YONH (Y is the same as defined above): [84]

[85] wherein, R , R , and Z are the same as defined above. [86] Reacting the compound formula (VII) with YONH may be carried out at about 40 0C without using a solvent. And also, the reaction may be carried out by refluxing in an organic solvent such as dichloromethane, acetonitrile, tetrahydrofuran, dimethyl sulfoxide, and toluene.

[87] The compound of formula (VII) may be obtained by a process which comprises reacting a compound of formula (VIII) with a compound of formula (IX): [88]

[89] [90]

[91] [92] wherein, R , R , and Z are the same as defined above and X is imidazolyl or N- methyl-N-methoxyamino.

[93] Reacting the compound of formula (VIII) with a compound of formula (IX) may be carried out in the presence of t-butyllithium. The reaction is carried out at about -78 ~ 0 0C in an organic solvent such as diethyl ether. [94] The compound of formula (VIII) may be obtained by a process which comprises reacting a compound of formula (X) with l,l'-carbonyldiimidazole. And also, the compound of formula (VIII) may be obtained by a process which comprises reacting a compound of formula (X) with isobutyl chloroformate, followed by reacting with N,O-dimethylhydroxyamine:

[95]

[96] wherein, R² and Z are the same as defined above.

[97]

[98] The present invention also provides an antibacterial composition comprising a therapeutically effective amount of a compound of formula (I) or its pharmaceutically acceptable salt and a pharmaceutically acceptable carrier. The composition of the present invention is useful for treating patients or animals with bacterial infections by administration or spreading over infected skins. The composition of the present invention is also useful for antibacterial purification and disinfection.

[99] The composition of the present invention may be orally or parenterally administered. The composition of the present invention for oral administration may be in the form of tablet, capsule, powder, granule, solution, suspension, or gel, and may comprise conventional additives such as a diluent, a disintegrating agent, and a lubricant. Examples of the additives include a diluent such as syrup, Arabic gum, gelatin, sorbitol, lactose, sugar, corn starch, calcium phosphate, glycine, magnesium stearate, talc, polyethylene glycol, silica, potato starch, and sodium lauryl sulfate, a flavorant, and a colorant. The composition of the present invention for parenteral administration (e.g., injection) may be an isotonic solution or may be sterilized. In this case, the composition of the present invention may comprise conventional additives such as a preservative and a stabilizer.

[100] The composition of the present invention may be administered in an average adult

(about 70 kg) dosage of about 7 mg/day to 35 g/day for antibacterial treatment. However, the dosage may be changed according to the type and the degree of severity of diseases. In this regard, a unit dosage form may comprise about 0.7 mg to 2.8 g of the compound of the present invention together with a pharmaceutically acceptable carrier.

[101]

[102] The following Examples are given for the purpose of illustration only, and are not intended to limit the scope of the invention. [103] [104] Example 1. pyridin-2-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]

- 1 -isopropyl-2-oxo-heptyl } -amide

[105] The titled compound was prepared according to the following Scheme 1:

[106] Scheme 1

[107]

[108] [109] Step 1. [(S)-l-(methoxy-methyl-carbamoyl)-2-methyl-propyl]-carbamic acid t-butyl ester

[HO] [111] The solution of N-Boc-L- valine (12 g, 55.23 mmol) in a mixed solvent (150 ml) of tetrahydrofuran (30 ml) and dichloromethane (120 ml) was cooled to -20 °C. 1-Methylpiperidine (6.7 ml, 55.23 mmol) and isobutyl chloroformate (7.54 ml, 55.23 mmol) were added dropwise to the cooled solution, which was then stirred for 5 minutes. N,O-dimethylhydroxyamine hydrochloride (5.38 g, 55.23 mmol) was suspended in dichloromethane (15 ml) and then 1-methylpiperidine (6.7 ml, 55.23 mmmol) was added dropwise thereto. The resulting solution of N,O-dimethylhydroxyamine in dichloromethane was added dropwise at -20 ⁰C to the reaction mixture, which was then stirred for 2 hours at room temperature. Water was added to the reaction mixture, which was then extracted with dichloromethane. The resulting organic layer was washed successively with 10% citric acid, a saturated NaHCO solution, and brine. The organic layer was dehydrated with anhydrous sodium sulfate and then dried under reduced pressure to obtain the titled compound (11 g, 76 %).

[112] ¹H-NMR(CDCl₃) δ 0.89 (d, 3H, J=6.8Hz), 0.94 (d, 3H, J=6.8Hz), 1.43 (s, 9H),

1.97 (m, IH), 3.21 (s, 3H), 3.77 (s, 3H), 4.52 (bs, IH), 5.2 (bs, IH).

[113]

[114] Step 2. ((S)-l-isopropyl-3-methylen-2-oxo-heptyl)-carbamic acid t-butyl ester

[115] The mixed solution of diethyl ether (60 ml) and t-butyllithium (1.7 M / pentane,

22.35 ml, 37.99 mmol) was cooled to -78 ⁰C and a solution of 2-bromohexene (2.16 ml, 15.2 mmol) in diethyl ether (10 ml) was added thereto. The reaction mixture was stirred at 0 ⁰C for 2 hours and then cooled to -78 °C. The solution of [(S)-l-(methoxy-methyl-carbamoyl)-2-methyl-propyl]-carbamic acid t-butyl ester (Ig, 3.8 mmol) in diethyl ether (10 ml) was added dropwise to the reaction mixture, which was stirred at -78 ⁰C for 1 hour and then at 0⁰C for 1 hour. Brine was added to the reaction mixture. The separated organic layer was dehydrated with anhydrous sodium sulfate and then concentrated under reduced pressure. The concentrate was purified with silica gel column chromatography (ethyl acetate : hexane = 1:10) to obtain the titled compound (1 g, 90 %).

[116] ¹H-NMR(CDCl₃) δ 0.75 (d, 3H, J=6.8Hz), 0.9 (t, 3H, J=7.1Hz), 0.98 (d, 3H,

J=6.8Hz), 1.37 (m, 4H), 1.44 (s, 9H), 2.05 (m, IH), 2.26 (m, 2H), 4.93 (m, IH), 5.23 (bs, IH), 5.83 (s, IH), 6.08 (s, IH).

[117]

[118] Step 3. [(lS,3R)-3-(benzyloxyamino-methyl)-l-isopropyl-2-oxo-heptyl]-carbamic acid t-butyl ester

[119] O-benzylhydroxyamine (1.62 g, 13.23 mmol) was added to

((S)-l-isopropyl-3-methylen-2-oxo-heptyl)-carbamic acid t-butyl ester (2.5 g, 8.82 mmol) and then stirred at 40 ⁰C for 12 hours. The reaction mixture was purified with silica gel column chromatography (ethyl acetate : hexane = 1:7) and each optical isomers were separated with silica gel thin layer chromatography (the polarity of (lS,3R)-optical isomer is slightly higher) to obtain the titled compound (2.1 g, 58 %).

[120] ¹H-NMR(CDCl₃) δ 0.74 (d, 3H, J=6.8Hz), 0.87 (t, 3H, J=6.9Hz), 1.10 (d, 3H,

J=6.8Hz), 1.25 (m, 4H), 1.43 (m, 10H), 1.50 (m, IH), 2.17 (m, IH), 2.93 - 3.03 (m, 2H), 3.20 (m, IH), 4.40 (m, IH), 4.63 (s, 2H), 5.09 (m, IH), 5.56 (bs, IH), 4.63 (s, 2H), 5.09 (m, IH), 5.56 (bs, IH), 7.28 - 7.37 (m, 5H).

[121]

[122] Step 4. {(lS,3R)-3-[(benzyloxy-formyl-amino)-methyl] - l-isopropyl-2-oxo-heptylJ-carbamic acid t-butyl ester

[123] A mixture of acetic anhydride (0.977 ml, 10.33 mmol) and formic acid (1.17 ml,

30.99 mmol, 96%) was stirred for 30 minutes and then added at room temperature to a solution of [(lS,3R)-3-(benzyloxyamino-methyl)-l-isopropyl-2-oxo-heptyl]-carbamic acid t-butyl ester (1.68 g, 4.13 mmol) in ethyl acetate (13 ml). The reaction mixture was stirred at room temperature for 1 hour and then washed with a saturated NaHCO solution. The separated organic layer was dehydrated with anhydrous sodium sulfate and then concentrated under reduced pressure. The concentrate was purified with silica gel column chromatography (ethyl acetate : hexane = 1:2) to obtain the titled compound (1.7 g, 95 %).

[124] ¹H-NMR(CDCl₃) δ 0.66 (d, 3H, J=6.6Hz), 0.87 (t, 3H, J=6.9Hz), 0.98 (m, 3H,

J=6.8Hz), 1.24 - 1.57 (m, 15H), 2.01 (bs, IH), 3.08 - 3.76 (m, 3H), 4.37 (m, IH), 4.78 - 5.05 (m, 3H), 7.34 (m, 5H).

[125]

[126] Step 5. N-

((2R,4S)-4-amino-2-butyl-5-methyl-3-oxo-hexyl)-N-benzyloxy-formamide trifluo- roacetate

[127] Trifluoroacetic acid (3 ml, 39.12 mmol) was added to a solution of

{(IS ,3R)-3- [(benzyloxy-formyl-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -carbamic acid t-butyl ester (1.7 g, 3.91 mmol) in dichloromethane (20 ml). The reaction mixture was stirred at room temperature for 6 hours and then concentrated under reduced pressure. The product was used in the subsequent step without further purification.

[128]

[129] Step 6. Pyridin-2-carboxylic acid {(lS,3R)-3-[(benzyloxy-formyl-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide

[ 130] 1-Hydroxybenzotriazole hydrate (0.316 g, 2.32 mmol), N- ethyl-N'-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.448 g, 2.32 mmol), N,N-diisopropylethylamine (0.93 ml, 5.42 mmol), and pyridin-2-carboxylic acid (0.194 g, 1.57 mmol) were added to a solution of N-

((2R,4S)-4-amino-2-butyl-5-methyl-3-oxo-hexyl)-N-benzyloxy-formamide trifluo- roacetate (0.67 g, 1.55 mmol) in dichloromethane (10 ml). The reaction mixture was stirred at room temperature for 12 hours and then washed with water. The separated organic layer was dehydrated with anhydrous sodium sulfate and then concentrated under reduced pressure. The concentrate was purified with silica gel column chromatography (ethyl acetate : hexane = 1:2) to obtain the titled compound (0.27 g, 39 %).

[131] ¹H-NMR(CDCl₃) δ 0.75 - 0.89 (m, 6H), 1.02 (m, 3H), 1.25 (m, 4H), 1.45 - 1.60 (m,

2H), 2.28 (m, IH), 2.94 - 3.23 (m, 3H), 4.85 (m, 2H), 6.15 (m, IH), 7.26 - 7.38 (m, 6H), 7.85 (m, IH), 8.15 (m, IH), 8.60 (m, IH).

[132]

[133] Step 7. Pyridin-2-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] -

1 -isopropyl-2-oxo-heptyl } -amide [ 134] Pd/C (0.019 g, 0.02 mmol, 10 wt%) was added to a solution of pyridin-2-carboxylic acid {(lS,3R)-3-[(benzyloxy-formyl-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide (0.16 g, 0.36 mmol) in methanol (3 ml). The reaction mixture was stirred at room temperature for 2 hours under hydrogen atmosphere. The reaction mixture was filtered to discard Pd/C and the filtrate was concentrated under reduced pressure. The concentrate was purified with silica gel column chromatography (dichloromethane : methanol = 15:1) to obtain the titled compound (0.066 g, 44 %).

[135] ¹H-NMR(CDCl₃) δ 0.83-1.08 (m, 9H), 1.25-1.50 (m, 5H), 1.70 (m, IH), 2.38 (m,

IH), 3.48 (m, 2H), 3.98 (m, IH), 4.93 (m, IH), 7.45 (m, IH), 7.86 (m, IH), 8.16 (m, IH), 8.60 (m, IH).

[136]

[137] The compounds of the following Examples were prepared in the same manner as in

Example 1, except that 6-methyl-pyridin-2-carboxylic acid, 4-chloro-pyridin-2-carboxylic acid, quinolin-2-carboxylic acid, isoquinolin- 1-carboxylic acid, 6-fluoro-pyridin-2-carboxylic acid, 3-methyl-pyridin-2-carboxylic acid, pyridin-3-carboxylic acid, pyridin-4-carboxylic acid, 3-fluoro-pyridin-2-carboxylic acid, 5-methyl-pyridin-2-carboxylic acid, 4-methoxy-pyridin-2-carboxylic acid, 5-fluoro-pyridin-2-carboxylic acid, 5-methyl-pyridin-3-carboxylic acid, 2-methoxy-pyridin-3-carboxylic acid, 2-fluoro-pyridin-3-carboxylic acid, 6-fluoro-pyridin-3-carboxylic acid, S-fluoro-pyridin-S-carboxylic acid, 2-methyl-pyridin-3-carboxylic acid, 4-methyl-pyridin-3-carboxylic acid, 2-methyl-pyridin-4-carboxylic acid, 3-methyl-pyridin-4-carboxylic acid, 2-cyano-pyridin-4-carboxylic acid, 2-methoxy-pyridin-4-carboxylic acid, 2-fluoro-pyridin-4-carboxylic acid, 3-fluoro-pyridin-4-carboxylic acid, phenyl-carboxylic acid, 3-methyl-phenyl-carboxylic acid, 2-fluoro-phenyl-carboxylic acid, 4-methyl-phenyl-carboxylic acid, 3-methoxy-phenyl-carboxylic acid, 3-fluoro-phenyl-carboxylic acid, or isopropyl-carboxylic acid was used instead of pyridin-2-carboxylic acid in Step 6 of Example 1.

[138]

[139] Example 2. 6-methyl-pyridin-2-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide

[140] ¹H-NMR(CDCl₃) δ 0.83-1.07 (m, 9H), 1.25-1.47 (m, 5H), 1.69 (m, IH), 2.37 (m

IH), 2.60 (s, 3H), 3A9 (m, 2H), 4.02 (m, IH), 4.93 (m, IH), 7.34 (m, IH), 7.72 (m, 3H), 7.98 (m, 3H), 8.66 (m, IH).

[141]

[142] Example 3. 4-chloro-pyridin-2-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [143] ¹H-NMR(CDCl₃) δ 0.79-1.09 (m, 9H), 1.23-1.43 (m, 5H), 1.60 (m, IH), 2.35 (m

IH), 2.95-3.38 (m, 3H), 4.95 (m, IH), 7.48 (m, IH), 7.88 (m, IH), 8.20 (m, IH), 8.60

(m, IH). [144] [145] Example 4. quinolin-2-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-atnino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [146] ¹H-NMR(CDCl ) δ 0.81-0.91 (m, 6H), 1.10 (m, 3H), 1.25-1.50 (m, 5H), 1.73 (m,

IH), 2.45 (m,lH), 3.50 (m, 2H), 4.03 (m, IH), 5.02 (m, IH), 7.63 (m, IH), 7.79 (m,

3H), 7.83 (m, 2H), 8.17-8.33 (m, 3H), 8.22 (m, IH). [147] [148] Example 5. isoquinolin- 1 -carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [149] ¹H-NMR(CDCl₃) δ 0.81-0.92 (m, 6H), 1.10 (m, 3H), 1.34-1.68 (m, 6H), 2.41 (m,

IH), 3.48 (m, 2H), 3.98 (m, IH), 5.00 (m, IH), 7.65-7.86 (m, 4H), 8.54 (m, IH), 8.80

(m, IH), 9.53 (m, IH). [150] [151] Example 6. 6-fluoro-pyridin-2-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [152] ¹H-NMR(CDCl ) δ 0.82-0.90 (m, 6H), 1.06 (m, 3H), 1.25-1.31 (m, 4H), 1.46 (m,

IH), 1.70 (m, IH), 2.38 (m, IH), 3.49 (m, 2H), 3.99 (m, IH), 4.91 (m, IH), 7.11 (m,

IH), 87.92-8.23 (m, 3H). [153] [154] Example 7. 3-methyl-pyridin-2-carboxylic acid

{(lS,3R)-3-[(foπnyl-hydroxy-amino)-πiethyl]-l-isopropyl-2-oxo-heptyl}-aπiide [155] ¹H-NMR(CDCl₃) δ 0.82-0.91 (m, 6H), 1.05 (m, 3H), 1.27 (m, 4H), 1.47 (m, IH),

1.68 (m, IH), 2.38 (m, IH), 2.71 (s, 3H), 3.46 (m, 2H), 3.97 (m, IH), 4.90 (m, IH),

7.31 (m, IH), 7.60 (m, IH), 8.43 (m, IH), 8.69 (m, IH). [156] [157] Example 8. pyridin-3-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]

- 1 -isopropyl-2-oxo-heptyl } -amide [158] ¹H-NMR(CDCl₃) δ 0.77-0.92 (m, 6H), 1.08 (m, 3H), 1.32 (m, 4H), 1.50 (m, IH),

1.74 (m, IH), 2.36 (bs, IH), 3.48 (m, 2H), 4.00 (m, IH), 5.04 (m, IH), 7.00 (m, IH),

7.43 (m, IH), 8.17 (m, IH), 8.74 (m, IH), 9.07 (m, IH). [159] [160] Example 9. pyridin-4-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]

- 1 -isopropyl-2-oxo-heptyl } -amide [161] ¹H-NMR(CDCl₃) δ 0.76-0.92 (m, 6H), 1.08 (m, 3H), 1.32 (m, 4H), 1.51 (m, IH),

1.68 (m, IH), 2.35 (m, IH), 3.50 (m, 2H), 4.00 (m, IH), 5.02 (m, IH), 6.85 (m, IH), 7.67 (d, 2H, J=5.3Hz), 8.78 (d, 2H, J=4.7Hz). [162]

[163] Example 10. 3-fluoro-pyridin-2-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [164] ¹H-NMR(CDCl₃) δ 0.80-0.95 (m, 6H), 1.07 (m, 3H), 1.26 (m, 4H), 1.45 (m, IH),

1.68 (m, IH), 2.37(m,lH), 3.46 (m, 2H), 3.99 (m, IH), 4.95 (m, IH), 7.48-7.64 (m, 2H), 8.38-8.45 (m, 2H).

[165]

[ 166] Example 11. 5-methyl-pyridin-2-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [167] ¹H-NMR(CDCl ) δ 0.82-1.06 (m, 9H), 1.30 (m, 4H), 1.45 (m, IH), 1.68 (m, IH),

2.40 (m, IH), 3.48 (m, 2H), 3.97 (m, IH), 4.91 (m, IH), 7.65 (m, IH), 8.60 (m, IH),

8.41 (s, IH), 8.55 (m, IH). [168]

[169] Example 12. 4-methoxy-pyridin-2-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [170] ¹H-NMR(CDCl₃) δ 0.82-1.07 (m, 9H), 1.25-1.46 (m, 5H), 1.70 (m, IH), 2.36 (m,

IH), 3.45 (m, 2H), 3.93 (m, 4H), 4.89 (m, IH), 6.94 (m, IH), 7.70 (s, IH), 8.38 (m,

IH), 8.63 (m, IH). [171] [172] Example 13. 5-fluoro-pyridin-2-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [173] ¹H-NMR(CDCl₃) δ 0.81-0.85 (m, 6H), 1.05 (m, 3H), 1.31 (m, 4H), 1.45 (m, IH),

1.68 (m, IH), 2.37 (m,lH), 3.47 (m, 2H), 3.97 (m, IH), 4.90 (m, IH), 7.56 (m, IH), 8.20 (m, 3H), 8.84 (m, 2H).

[174]

[175] Example 14. 5-methyl-pyridin-3-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [176] ¹H-NMR(CDCl₃) δ 0.77-0.89 (m, 6H), 1.03 (m, 3H), 1.32 (m, 4H), 1.50 (m, IH),

1.69 (m, IH), 2.40 (s, 4H), 3.50 (m, 2H), 4.00 (m, IH), 5.02 (m, IH), 7.80 (m, IH), 8.52 (m, IH), 8.84 (m, IH).

[177]

[178] Example 15. 2-methoxy-pyridin-3-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [179] ¹H-NMR(CDCl₃) δ 0.80 (m, 6H), 1.02 (m, 3H), 1.08-1.46 (m, 5H), 1.72(m, IH),

2.37 (m, IH), 3.47 (m, 2H), 3.97 (m, IH), 4.15 (s, 3H), 4.98 (m, IH), 7.07 (m, IH),

8.29 (m, IH), 8.47 (m, IH). [180] [181] Example 16. 2-fluoro-pyridin-3-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [182] ¹H-NMR(CDCl₃) δ 0.79-0.92 (m, 6H), 1.08 (m, 3H), 1.33 (m, 4H), 1.50 (m, IH),

1.70 (m, IH), 2.38 (m, IH), 3.48 (m, 2H), 3.99 (m, IH), 5.00 (m, IH), 7.34 (m, 2H),

8.36 (s, IH), 8.53 (m, IH). [183] [184] Example 17. 6-fluoro-pyridin-3-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [185] ¹H-NMR(CDCl₃) δ 0.77-1.07 (m, 9H), 1.33 (m, 4H), 1.47 (m, IH), 1.65 (m, IH),

2.35 (bs, IH), 3.50 (m, 2H), 3.99 (m, IH), 5.03 (m, IH), 6.98 (m, 2H), 8.26 (bs, IH),

8.68 (m, IH). [186] [187] Example 18. 5-fluoro-pyridin-3-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [188] ¹H-NMR(CDCl₃) δ 0.77-0.93 (m, 6H), 1.08 (m, 3H), 1.33 (m, 4H), 1.50 (m, IH),

1.70 (m, IH), 2.36 (bs IH), 3.50 (m, 2H), 3.99 (m, IH), 5.02 (s, IH), 6.90 (bs, IH),

7.88 (s, 3H), 8.62 (s,lH), 8.86 (m, IH). [189] [190] Example 19. 2-methyl-pyridin-3-carboxylic acid

{(lS,3R)-3-[(foπnyl-hydroxy-amino)-πiethyl]-l-isopropyl-2-oxo-heptyl}-amide [191] ¹H-NMR(CDCl₃) δ 0.73-0.94 (m, 6H), 1.06 (m, 3H), 1.36 (m, 4H), 1.54 (m, IH),

1.72 (m, IH), 2.33 (m,lH), 2.67(s, 3H), 3.50 (m, 2H), 4.00 (m, IH), 5.01 (m, IH), 7.18 (m, IH), 7.70 (m, 3H), 8.58 (s, IH).

[192]

[193] Example 20. 4-methyl-pyridin-3-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [194] ¹H-NMR(CDCl ) δ 0.74-0.93 (m, 6H), 1.10 (m, 3H), 1.35 (m, 4H), 1.55 (m, IH),

1.73 (m, IH), 2.34 (m, IH), 2.47 (s, 3H), 3.48 (m, 2H), 4.01 (m, IH), 5.02 (m, IH), 7.20 (m, IH), 8.50 (m, IH), 8.63 (s, IH).

[195]

[196] Example 21. 2-methyl-pyridin-4-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [197] ¹H-NMR(CDCl₃) δ 0.72-0.93 (m, 6H), 1.09 (m, 3H), 1.34(m, 4H), 1.45(m, IH),

1.68(m, IH), 2.34(m, IH), 2.42 (s, 3H), 3.47 (m, 2H), 3.98 (m, IH), 4.98 (m, IH), 6.79

(m, IH), 7.34 (m, IH), 7.85(m, IH), 8.50(m, 2H). [198] [199] Example 22. 3-methyl-pyridin-4-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [200] ¹H-NMR(CDCl₃) δ 0.75-0.92 (m, 6H), 1.06 (m, 3H), 1.31 (m, 4H), 1.50 (m, IH),

1.68 (m, IH), 2.35 (m, IH), 2.66(s, 3H), 3.47 (m, 2H), 3.98 (m, IH), 4.99 (m, IH),

7.06 (m, IH), 7.53 (m, 2H), 7.86(m, IH), 8.64 (m, 2H). [201] [202] Example 23. 2-cyano-pyridin-4-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-atnino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [203] ¹H-NMR(CDCl ) δ 0.75-0.90 (m, 6H), 1.07 (m, 3H), 1.62(m, 4H), 2.01(m, IH),

2.34(m, IH), 3.47 (m, 2H), 3.98 (m, IH), 7.33 (m, 3H), 7.87 (m, IH), 8.08(m, IH),

8.87(m, IH). [204] [205] Example 24. 2-methoxy-pyridin-4-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [206] ¹H-NMR(CDCl₃) δ 0.74-0.91 (m, 6H), 1.05 (m, 3H), 1.31 (m, 4H), 1.48 (m, IH),

1.66 (m, IH), 2.33 (m, IH), 3.48 (m, 2H), 4.00 (m, 4H), 4.98 (m, IH), 6.72 (m, IH),

7.08 (m, IH), 7.19 (m, IH), 7.84 (m, IH), 8.27 (m, 2H). [207] [208] Example 25. 2-fluoro-pyridin-4-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-atnino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [209] ¹H-NMR(CDCl₃) δ 0.74-0.92 (m, 6H), 1.06 (m, 3H), 1.49 (m, 4H), 1.67(m, IH),

2.36(m, IH), 3.48 (m, 2H), 3.98 (m, IH), 5.00 (m, IH), 6.91 (m, IH), 7.36 (m, 2H),

7.54 (m, IH), 7.85 (m, IH), 8.36(m, IH). [210] [211] Example 26. 3-fluoro-pyridin-4-carboxylic acid

{(lS,3R)-3-[(foπnyl-hydroxy-amino)-πiethyl]-l-isopropyl-2-oxo-heptyl}-aπiide [212] ¹H-NMR(CDCl₃) δ 0.76-0.90 (m, 6H), 1.09 (m, 3H), 1.34(m, 4H), 1.45(m, IH),

1.64(m, IH), 2.36(m, IH), 3.47 (m, 2H), 3.98 (m, IH), 5.01 (m, IH), 7.19 (m, IH),

7.89 (m, 2H), 8.60 (m, 2H). [213] [214] Example 27. phenyl-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]

1 -isopropyl-2-oxo-heptyl } -amide [215] ¹H-NMR(CDCl₃) δ 0.76-0.90 (m, 6H), 1.07 (m, 3H), 1.31 (m, 4H), 1.49 (m, IH),

1.68 (m, IH), 2.34 (bs, IH), 3.47 (m, 2H), 3.97 (m, IH), 5.04 (m, IH), 6.72 (m, IH),

7.50 (m, 3H), 7.82 (m, 3H). [216] [217] Example 28. 3-methyl-phenyl-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [218] ¹H-NMR(CDCl₃) δ 0.77-0.91 (m, 6H), 1.02-1.08 (m, 3H), 1.33(m, 4H), 1.45(m,

IH), 1.68(m, IH), 2.35 (m, IH), 2.41 (s, 3H), 3.48 (m, 2H), 3.96 (m, IH), 5.01 (m, IH), 6.67 (m, IH), 7.34 (m, 2H), 7.59(m,2H),7.84 (s, IH). [219] [220] Example 29. 2-fluoro-phenyl-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [221] ¹H-NMR(CDCl₃) δ 0.77-0.91 (m, 6H), 1.06-1.14 (m, 3H), 1.33(m, 4H), 1.45(m,

IH), 1.70(m, IH), 2.35 (m, IH), 3.47 (m, 2H), 3.98 (m, IH), 5.02(m, IH), 7.12-7.28

(m, 3H), 7.48 (m, IH), 7.84 (s, IH) ,8.04(m, IH). [222] [223] Example 30. 4-methyl-phenyl-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [224] ¹H-NMR(CDCl ) δ 0.66-0.90 (m, 6H), 1.05-1.07 (m, 3H), 1.31(m, 4H), 1.45(m,

IH), 1.68(m, IH), 2.32(m, IH), 2.40 (s, 3H), 3.50 (m, 2H), 3.97 (m, IH), 5.01 (m, IH),

6.65 (m, IH), 7.320-7.29 (m, 2H), 7.69(m,2H), 7.85 (s, IH). [225] [226] Example 31. 3-methoxy-phenyl-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [227] ¹H-NMR(CDCl₃) δ 0.76-0.91 (m, 6H), 1.05-1.08 (m, 3H), 1.33(m, 4H), 1.48(m,

IH), 1.68(m, IH), 2.33 (m, IH), 3.48 (m, 2H),3.85(s, 3H), 3.97 (m, IH), 5.00 (m, IH),

6.68(m, IH), 7.05(m, IH), 7.35(m,2H),7.84 (s, IH). [228] [229] Example 32. 3-fluoro-phenyl-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [230] ¹H-NMR(CDCl₃) δ 0.73-0.91 (m, 6H), 1.01-1.08 (m, 3H), 1.31(m, 4H), 1.48(m,

IH), 1.68(m, IH), 2.33 (m, IH), 3.48 (m, 2H), 3.97 (m, IH), 5.01 (m, IH), 6.70 (m,

IH), 7.22 (m, IH), 7.42(m, IH), 7.55(m, 2H), 7.84 (s, IH). [231] [232] Example 33. isopropyl-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide [233] ¹H-NMR(CDCl₃) δ 0.69-0.99 (m, 9H), 1.14-1.25 (m, 6H), 1.30(m, 4H), 1.46(m,

IH), 1.65(m, IH), 2.21 (m, IH), 2.43 (m, IH), 3.46 (m, 2H), 3.96 (m, IH), 4.78 (m,

IH), 5.97 (m, IH), 7.82 (m, IH). [234]

[235] Experimental Example 1 : Evaluation of deformylase inhibitory activity

[236]

[237] ( 1 ) Production and purification of deformylase

[238] [239] Deformylase-producing bacteria strains (Novagen, America, cat. no. 69041) were inoculated into 30ml of LB media (tryptone 8g/l, yeast extract 5 g/1, NaCl 5 g/1, and IN NaOH 2.5 ml) containing ampicillin and then cultured at 37 ⁰C. The cultures were then inoculated into new LB media (3L) with the same composition as in the above- described LB media. When O.D (660 nm) reached 0.5, protein expression was induced by addition of 1 mM isopropyl-β-D-thiogalactopyranoside to the LB media. Then, cell pellets were harvested by centrifugation, incubated at -80 ⁰C for 30 minutes, re- suspended in a phosphate buffer, followed by lysis of bacteria cells by sonification with 8 seconds interval between each pulse (1 second). The supernatant was obtained by ultracentrifugation, loaded onto an affinity column, followed by elution with an elution buffer. The eluted fractions containing peptide deformylase were identified by SDS-PAGE, followed by elution by gel chromatography. At this time, a buffer containing 5 mM NiCl was used as a stabilizer for the enzyme source, and SDS- PAGE and Dynamic Light Scattering were used to identify the purification of the enzyme source. The purified enzyme source was kept at -80 °C until use.

[240] [241] (2) Deformylase inhibition test [242] [243] Each of the compounds prepared in Examples was dissolved in dimethyl sulfoxide to a concentration of 2 mM. 7D of each of the reaction mixtures was loaded into each well of microtiter plates. The deformylase prepared in section (1) was diluted with a buffer (5OmM HEPES, pH7.0, 1OmM NaCl, 5mM NiCl₂, 0.1% Triton X-100) to a concentration of 250 nM and then loaded into each well of the microtiter plates. Then, continuous dilution for each of the compounds of Examples was performed with a dilution ratio of 7:2 to make a final volume of 50 D for each well, followed by incubation at room temperature for 5 minutes. Then, 50 D of 5 mM formyl-Met-Ala-Ser used as a substrate source was loaded into each well and incubated at 30⁰C for 10 minutes to induce enzyme reaction. When the enzyme reaction was completed, the obtained samples were treated with 50 D of fluorescamine and 50 D of 50 mM borate- sodium hydroxide buffer (pH 9.5) to measure fluorescence at an excitation wavelength of 390 nm and an emission wavelength of 465 nm. Based on the fluorescence thus measured, the concentrations (IC ) of the samples that inhibit enzyme activity by 50% were calculated and the results are presented in Table 1 below.

[244] Table 1

[245] [246] Experimental Example 2: Antibacterial effect test [247] [248] Minimal inhibitory concentrations (MICs) of the compounds of Examples were determined for Hemophilus influenza (ATCC 51907), Streptococcus pneumoniae (ATCC 6305), and Moraxella cararrhalis (ATCC 43617).

[249] Each of the compounds of Examples was dissolved in dimethyl sulfoxide to a concentration of 2 mg/ml. At this time, actinonin (Sigma, America) or ampicillin (Sigma, America) was used as standard antibiotic in a concentration of 2 mg/ml.

[250] For bacteria culture, Hemophilus influenza and Moraxella cararrhalis were inoculated into culture media such as brain heart infusion (37 g/1, Difco, America) supplemented with NAD 10 mg/1 and hemin 5 mg/1 and then cultured for 24 and 48 hours, respectively. Streptococcus pneumoniae was inoculated into GC media containing 5% horse serum and cultured for 48 hours. The cultures were diluted to a concentration of 1 x 10 CFU/ml and inoculated into wells of microtiter plates. After incubation at 37°C, 5% CO for 24 hours for Hemophilus influenza and 48 hours for Moraxella cararrhalis and Streptococcus pneumoniae, MICs, the lowest concentrations to inhibit visual growth, were determined and the results are presented in Table 2 below.

[251] Table 2

[252]

Industrial Applicability

[253] The compounds of the present invention or the salts thereof have an excellent inhibitory activity against deformylase, thereby being useful for an antibacterial agent having a broad spectrum against bacteria, including bacteria with resistance to existing antibacterial agents.

Claims

[1] A compound of formula (L) or its pharmaceutically acceptable salt:

wherein, R is a straight or branched C ~C alkyl group,

R is hydrogen or a straight or branched C ~C alkyl group, and

3 . . ^{1 6}

R is a straight or branched C ~C alkyl group, a pyridinyl group, a phenyl

1 6 group, a quinolinyl group, or a isoquinolinyl group (wherein, the pyridinyl group, the phenyl group, the quinolinyl group, or the isoquinolinyl group may be substituted with 1 to 3 substituent(s) selected from the group consisting of C ~C

1 4 alkyl, C ~C alkoxy, halogen, and cyano).

1 4

[2] The compound of formula (I) or its pharmaceutically acceptable salt of claim 1, wherein R is a radical of formula (Ha), (lib), or (TIc):

wherein, R , R , R , and R , independently each other, hydrogen, a C ~C alkyl group, a C ~C alkoxy group, a halogen group, or a cyano group, and * is a

1 4 binding site.

[3] The compound of formula (I) or its pharmaceutically acceptable salt of claim 1, which is selected from the group consisting of: pyridin-2-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide;

6-methyl-pyridin-2-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl ]- 1 -isopropyl-2-oxo-heptyl } -amide;

4-chloro-pyridin-2-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide; quinolin-2-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide; isoquinolin-l-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide;

6-fluoro-pyridin-2-carboxylic acid { (lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide;

3-methyl-pyridin-2-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl ]- 1 -isopropyl-2-oxo-heptyl } -amide; pyridin-3-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide; pyridin-4-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide;

3-fluoro-pyridin-2-carboxylic acid { (lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide;

5-methyl-pyridin-2-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl ]- 1 -isopropyl-2-oxo-heptyl } -amide; 4-methoxy-pyridin-2-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide; 5-fluoro-pyridin-2-carboxylic acid { (lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide;

5-methyl-pyridin-3-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl ]- 1 -isopropyl-2-oxo-heptyl } -amide; 2-methoxy-pyridin-3-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide; 2-fluoro-pyridin-3-carboxylic acid { (lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide; ό-fluoro-pyridin-S-carboxylic acid { (lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide; S-fluoro-pyridin-S-carboxylic acid { (lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide;

2-methyl-pyridin-3-carboxylic acid {(lS,3R)-3-[(foπnyl-hydroxy-amino)-methyl ]- 1 -isopropyl-2-oxo-heptyl } -amide;

4-methyl-pyridin-3-carboxylic acid {(lS,3R)-3-[(foπnyl-hydroxy-amino)-methyl ]- 1 -isopropyl-2-oxo-heptyl } -amide;

2-methyl-pyridin-4-carboxylic acid {(lS,3R)-3-[(foπnyl-hydroxy-amino)-methyl ]- 1 -isopropyl-2-oxo-heptyl } -amide;

3-methyl-pyridin-4-carboxylic acid {(lS,3R)-3-[(formyl-hydτoxy-amino)-methyl ]- 1 -isopropyl-2-oxo-heptyl } -amide;

2-cyano-pyridin-4-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide; 2-methoxy-pyridin-4-carboxylic acid

{(lS,3R)-3-[(formyl-hydroxy-amino)-methyl]-l-isopropyl-2-oxo-heptyl}-amide; 2-fluoro-pyridin-4-carboxylic acid { (lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide;

3-fluoro-pyridin-4-carboxylic acid { (lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide; phenyl-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide;

3-methyl-phenyl-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide;

2-fluoro-phenyl-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide;

4-methyl-phenyl-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide;

3-methoxy-phenyl-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide;

3-fluoro-phenyl-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - l-isopropyl-2-oxo-heptyl}-amide; and isopropyl-carboxylic acid {(lS,3R)-3-[(formyl-hydroxy-amino)-methyl] - 1 -isopropyl-2-oxo-heptyl } -amide.

[4] A process for preparing a compound of formula (I) or its pharmaceutically acceptable salt, which comprises performing a deprotection reaction of a compound of formula (HI):

wherein, R¹, R², and R³ are the same as defined in claim 1 and Y is a hydroxy- protecting group.

[5] An antibacterial composition comprising a therapeutically effective amount of the compound of formula (I) or its pharmaceutically acceptable salt of any one of claims 1 to 4 and a pharmaceutically acceptable carrier.