KR100245983B1 - Novel quinolone carboxylic derivative - Google Patents

Abstract

The present invention relates to a novel quinolonecarboxylic acid derivative of formula (I) and a pharmaceutically acceptable salt thereof, a method for preparing the same, and an antimicrobial composition containing the same as an active ingredient, wherein the compound of formula (I) It is excellent as an antibacterial agent and has a broad antibacterial spectrum, which is useful as an antibacterial agent.

(Wherein X, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ and R ₆ are as defined in the specification).

Description

New Quinolone Carboxylic Acid Derivatives

The present invention relates to a novel quinolonecarboxylic acid derivative having excellent antimicrobial activity, a pharmaceutically acceptable salt thereof, a preparation method thereof, and an antimicrobial composition containing the same as an active ingredient.

The quinolone derivatives reported so far and the quinolone antimicrobial agents used in the clinic are known to have excellent antimicrobial activity, but the antimicrobial activity against gram-negative bacteria is inferior to that of gram-negative bacteria, or the gram-negative bacteria is excellent. There is a problem such as poor or showing cytotoxicity, phototoxicity and the like.

Accordingly, the present inventors have an excellent antimicrobial activity, a broad antimicrobial spectrum, an improved antimicrobial activity against quinone derivatives, particularly gram-positive bacteria and methicillin-resistant bacteria, with low solubility in water and low cytotoxicity. As a result of continuing research to develop an antimicrobial agent, a quinone derivative having a 3,4-substituted pyrrolidine derivative at position 7 of the quinolone mother core, prepared by condensing a quinolone carboxylic acid and an amine derivative, is not only Gram-negative but also Gram-positive And it was found that the antimicrobial activity against methicillin resistant strains is very good and the solubility in water is good and the selectivity is low, leading to the completion of the present invention.

Accordingly, an object of the present invention is to provide a quinolone carboxylic acid derivative having excellent antimicrobial activity and broad antibacterial spectrum, low solubility in water and low cytotoxicity, a preparation method thereof, and an antimicrobial composition comprising the same as an active ingredient.

In order to achieve the above object, the present invention provides a novel quinolonecarboxylic acid derivative of the general formula (I) and a pharmaceutically acceptable salt thereof:

Formula 1

In the above formula,

R ₁ means a C ₁ -C ₄ lower alkyl group such as ethyl, cyclopropyl, 1-t-butyl group, or a C ₁ -C ₄ lower alkyl group, phenyl group or pyridyl group substituted with halogen;

R ₂ , R ₃ , R ₄ and R ₆ each represent hydrogen, a C ₁ -C ₄ lower alkyl group, or a C ₁ -C ₄ lower alkyl group substituted with phenyl or halogen;

R ₅ means hydrogen, amino or methyl group;

X means nitrogen, methine group (CH) or substituted methine group (CZ), wherein Z is halogen, C ₁ -C ₄ lower alkyl group or alkoxy group, or C ₁ -C ₄ lower alkyl group substituted by halogen or An alkoxy group;

X and R ₁ may be connected to each other to form COCH ₂ CH (CH ₃ ), CCH ₂ CH ₂ CH (CH ₃ ), or CSCH ₂ CH (CH ₃ ).

According to this another object, the present invention provides a method for preparing a compound of formula (I) by condensation reaction of an amine derivative of formula (II) with a quinolone derivative of formula (III):

Wherein R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and X are as defined above.

In accordance with another object, the present invention provides an antimicrobial composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

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

Compounds of formula (I) of the invention include their racemic compounds, the respective enantiomers and mixtures thereof.

In the compound of formula (I), R ₁ is a C ₁ -C ₄ lower alkyl group such as ethyl, cyclopropyl, 1-t-butyl group, or a C ₁ -C ₄ lower alkyl group, phenyl group or pyridyl group substituted with halogen and, R _2, R _3, R ₄ and R ₆ are each hydrogen, a C ₁ -C ₄ lower alkyl group, or a lower alkyl group of C ₁ -C ₄ substituted with phenyl or halogen, R ₅ is hydrogen, amino or methyl And X is nitrogen, methine, halomethine, methoxymethine or methylmethine; R ₂ , R ₃ , R ₄ and R ₆ are each hydrogen, benzyl or methyl, R ₅ is hydrogen, X and R ₁ are COCOH ₂ CH (CH ₃ ), CCH ₂ CH ₂ CH (CH ₃ ) or CSCH Preferred are compounds in which ₂ CH (CH ₃ ) is linked to each other to form a ring.

Among the compounds of the formula (I), in particular R ₁ is cyclopropyl, ethyl, 2-fluoroethyl or 2,4-difluorophenyl, and R ₂ , R ₃ , R ₄ and R ₆ are each hydrogen, benzyl or Methyl, R ₅ is hydrogen or amino, X is nitrogen, methine, fluormethine, chloromethine or methoxymethine, and R ₂ , R ₃ , R ₄ and R ₆ are each hydrogen, benzyl or methyl, More preferably, a compound in which R ₅ is hydrogen and X and R ₁ are connected to each other by COCH ₂ CH (CH ₃ ) to form a ring.

Among the compounds of the general formula (I) of the present invention, the most preferred compounds are as follows:

1-cyclopropyl-6,8-difluoro-7-[(3-N-benzylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline-3 -Carboxylic acid;

1-cyclopropyl-6-fluoro-8-chloro-7-[(3-N-benzylamino-4-aminomethylpyrrodine) -1-yl] -4-oxo-1,4-dihydroquinoline-3 -Carboxylic acid;

1-cyclopropyl-6-fluoro-8-methoxy-7-[(3-N-benzylamino-4-aminomethylpyrrodine) -1-yl] -4-oxo-1,4-dihydroquinoline- 3-carboxylic acid;

1-cyclopropyl-5-amino-6,8-difluoro-7-[(3-N-benzylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-di Hydroquinoline-3-carboxylic acid;

1-cyclopropyl-6-fluoro-7-[(3-N-benzylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline-3-carboxyl mountain;

9-Fluoro-2,3-dihydro-3- (S) -methyl-10-[(3-N-benzylamino-4-aminomethylpyrrolidin) -1-yl] -7-oxo-7H- Pyrido [1.2.3-dec] -1,4-benzooxazine-6-carboxylic acid;

1-cyclopropyl-6-fluoro-7-[(3-N-benzylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydro-1,8-naphthy Lidine-3-carboxylic acid;

1- (2,4-difluorophenyl) -6-fluoro-7-[(3-N-benzylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-di Hydro-1,8-naphthyridine-3-carboxylic acid;

1-cyclopropyl-6,8-difluoro-7-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline-3-carboxyl mountain;

1-cyclopropyl-6-fluoro-8-chloro-7-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline-3-carbox Acid;

1-cyclopropyl-6-fluoro-8-methoxy-7-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline-3- Carboxylic acid;

1-cyclopropyl-5-amino-6,8-difluoro-7-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline- 3-carboxylic acid;

9-Fluoro-2,3-dihydro-3- (S) -methyl-10-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -7-oxo-7H-pyrido [ 1.2.3-de] -1,4-benzooxazine-6-carboxylic acid;

1-cyclopropyl-6-fluoro-7-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydro-1,8-naphthyridine-3 -Carboxylic acid;

1-cyclopropyl-6-fluoro-7-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline-3-carboxylic acid;

1-cyclopropyl-6,8-difluoro-7-[(3-N-methylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline-3 -Carboxylic acid;

1-cyclopropyl-6-fluoro-8-chloro-7-[(3-N-methylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline- 3-carboxylic acid;

1-cyclopropyl-6-fluoro-8-methoxy-7-[(3-N-methylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline 3-carboxylic acid;

1-cyclopropyl-5-amino-6,8-difluoro-7-[(3-N-methylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-di Hydroquinoline-3-carboxylic acid;

9-Fluoro-2,3-dihydro-3- (S) -methyl-10-[(3-N-methylamino-4-aminomethylpyrrolidin) -1-yl] -7-oxo-7H- Pyrido [1.2.3-dec] -1,4-benzooxazine-6-carboxylic acid;

1-cyclopropyl-6,8-difluoro-7-[(3-amino-4-N-methylaminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline-3 -Carboxylic acid;

1-cyclopropyl-6-fluoro-8-methoxy-7-[(3-amino-4-N-methylaminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline 3-carboxylic acid;

1-cyclopropyl-6,8-difluoro-7-[(3-N-methylamino-4-N-methylaminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydro Quinoline-3-carboxylic acid;

1-cyclopropyl-6,8-difluoro-7-{[3-N- (3'-pyridylmethyl) amino-4-aminomethylpyrrolidin] -1-yl} -4-oxo-1, 4-dihydroquinoline-3-carboxylic acid;

1-cyclopropyl-6,8-difluoro-7-{[3-amino-4-N- (2-fluoroethyl) aminomethylpyrrolidin] -1-yl} -4-oxo-1,4- Dihydroquinoline-3-carboxylic acid;

1-cyclopropyl-6-fluoro-8-methoxy-7-{[3-amino-4-N- (2-fluoroethyl) aminomethylpyrrolidin] -1-yl} -4-oxo-1, 4-dihydroquinoline-3-carboxylic acid;

1-cyclopropyl-6-fluoro-7-{[3-amino-4-N- (2-fluoroethyl) aminomethylpyrrolidin] -1-yl} -4-oxo-1,4-dihydro- 1,8-naphthyridine-3-carboxylic acid;

1-cyclopropyl-6,8-difluoro-7-{[3-N- (2-fluoroethyl) amino-4-aminomethylpyrrolidin] -1-yl} -4-oxo-1,4- Dihydroquinoline-3-carboxylic acid;

1-cyclopropyl-6-fluoro-8-methoxy-7-{[3-N- (2-fluoroethyl) amino-4-aminomethylpyrrolidin] -1-yl} -4-oxo-1, 4-dihydroquinoline-3-carboxylic acid;

1-cyclopropyl-6-fluoro-7-{[3-N- (2-fluoroethyl) amino-4-aminomethylpyrrolidin] -1-yl} -4-oxo-1,4-dihydro- 1,8-naphthyridine-3-carboxylic acid;

1-cyclopropyl-6-fluoro-7-{[3-methoxyamino-4-aminomethylpyrrolidin] -1-yl} -4-oxo-1,4-dihydro-1,8-naphthyridine 3-carboxylic acid; And

1-cyclopropyl-6-fluoro-8-methoxy-7-{[3-amino-4- (N-methoxy) aminomethylpyrrolidin] -1-yl} -4-oxo-1,4- Dihydroquinoline-3-carboxylic acid.

The compound of general formula (I) according to the present invention can be prepared by condensation reaction of an amine derivative of general formula (II) with a quinolonecarboxylic acid of general formula (III), as shown in Scheme (1) below. :

Wherein R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and X are as defined above.

In the condensation reaction of Scheme 1, the compound of Formula (II) and the compound of Formula (III) are dissolved or suspended in water or an organic solvent in a molar ratio of 1: 1 to 1.5: 1, and an inorganic or organic base is added thereto. After the addition, it may be carried out by reacting at a temperature of 20 to 120 ℃. In addition, their racemic compounds, each enantiomer, can be prepared separately.

In this case, acetonitrile, dimethylformamide, dimethyl sulfoxide, pyridine, and the like may be used as the organic solvent, and triethylamine, pyridine, diazabicyclo [5,4,0] undec-7-ene, Tertiary amines, such as diisopropylamine, are suitable.

The amine derivatives of formula (II) used as starting materials for the preparation of compounds of formula (I) of the present invention are prepared according to the methods described in Tetrahedron Letters , 132, 1565-1568 (1991). It can be prepared as shown in Schemes 2 to 12, and their racemic compounds, each enantiomer can be prepared separately.

Wherein Bn is a benzyl group, BOC is a t-butoxycarbonyl group, Ms is a methanesulfonyl group, and Me is a methyl group.

Compounds of the general formula (III) used as preparations for the preparation of the compounds of general formula (I) of the present invention are described in Chem. Pharm. Bull. , 34, 4098 (1986); J. Med. Chem. , 34, 168 (1991); J. Med. Chem. , 23, 1358 (1980); J. Med. Chem. , 31, 503 (1988); Japanese Patent Laid-Open No. 62-25772; European Patent Application Publication No. 0 183 129 A1; And PCT International Publication No. WO 95/10519.

Pharmaceutically acceptable salts of the compounds of general formula (I) of the invention include their alkali metal and alkaline earth metal salts, such as sodium salts, potassium salts, magnesium salts and calcium salts.

Compounds of formula (I) or pharmaceutically acceptable salts thereof of the present invention are useful as antibacterial agents. The present invention therefore also provides an antimicrobial composition comprising an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

The pharmaceutical compositions of the invention can be formulated in oral or injection dosage forms. Formulations for oral administration include, for example, tablets, capsules, etc. These formulations, in addition to the active ingredient, may contain diluents (e.g. lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and / or glycine), lubricants ( Eg silica, talc, stearic acid and its magnesium or calcium salts and / or polyethylene glycols). Tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpicolidine, optionally starch, agar, alginic acid or its Disintegrants or boiling mixtures such as sodium salts and / or absorbents, colorants, flavors and sweeteners may be contained. Injectable formulations are preferably aqueous isotonic solutions or suspensions.

These formulations may be sterile and / or contain preservatives, stabilizers, hydrating or emulsifying accelerators, auxiliaries such as salts and / or buffers for osmotic pressure control and other therapeutically valuable substances.

The compositions of the present invention may be formulated by conventional mixing, granulating or coating methods and the active ingredient may be contained in the range of about 1 to 99.5%, preferably about 5 to 95%. The unit dosage form for about 1 to 300 kg mammals contains about 20 to 300 mg of active ingredient.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these preparation examples and examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.

Preparation Example 1: Preparation of trans-3-amino-4-aminomethylpyrrolidine trichloride

Step 1) Preparation of trans-N-benzyl-3-benzylamino-4-hydroxymethylpyrrolidine

To a suspension of 1.5 g of lithium aluminum hydride in 100 ml of tetrahydrofuran was slowly added dropwise a solution of 12.98 g of trans-N-benzyl-3-ethyloxycarbonyl-4-benzylaminopyrrolidine dissolved in 60 ml of tetrahydrofuran. . After 5 hours, the reaction was terminated with water, and the white solid was filtered off. The solvent was evaporated under reduced pressure, extracted with ethyl acetate, dried over anhydrous magnesium sulfate, and distilled under reduced pressure to obtain 11 g of the target compound.

¹ H NMR (CDCl ₃ ): δ 2.11 (1H, m), 2.30 (1H, q), 2.50 (1H, q) 2.73 (1H, q), 2.8 (2H, br s), 3.00 (1H, q) , 3.21 (1 H, m), 3.5-3.72 (6 H, m), 7.27 (10 H, m).

Step 2) Preparation of trans-N- (t-butyloxycarbonyl) -3- (N-t-butyloxycarbonyl) amino-4-methanesulfonyloxymethylpyrrolidine

3.5 g of trans-N-benzyl-3-benzylamino-4-hydroxymethylpyrrolidine was dissolved in 40 ml of dichloromethane, 1.56 ml of triethylamine was added, cooled to -30 ° C, and then methanesulfonyl chloride 1.02 ml was slowly added dropwise. After 30 minutes of reaction, water was added, the reaction was terminated, extracted with ethyl acetate, washed with water again, dried over anhydrous magnesium sulfate, and the solvent was evaporated under reduced pressure. The residue was dissolved in 30 ml of methanol, and 4 g of di-t-butyldicarbonate and 2.1 g of 10% Pd-C were added thereto. After hydrogenation for 14 hours, the catalyst was filtered off and the solvent was evaporated under reduced pressure. The residue was subjected to silica gel column chromatography (eluent: ethyl acetate: hexane = 1: 4) to obtain 4.3 g of the target compound.

¹ H NMR (CDCl ₃ ): δ1.42 (18H, s), 2.45 (1H, q), 3.0-3.3 (5H, m), 3.5-3.8 (2H, m), 3.9-4.36 (3H, m) , 4.82 (1 H, d).

Step 3) Preparation of trans-N- (t-butyloxycarbonyl) -3- (N-t-butyloxycarbonyl) amino-4-azidomethylpyrrolidine

1 g of trans-N- (t-butyloxycarbonyl) -3- (Nt-butyloxycarbonyl) amino-4- (methanesulfonyl) oxymethylpyrrolidine is dissolved in 20 ml of dimethylformamide, and here After adding 0.83 g of sodium azide to the mixture, the mixture was reacted at 60 DEG C overnight, water was added thereto, and extracted with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate, the organic solvent was distilled under reduced pressure, and silica gel column chromatography (eluate: ethyl acetate: hexane = 2: 1) gave 0.714 g of the target compound.

¹ H NMR (CDCl ₃ ): δ 1.43 (18H, s), 2.2 (1H, m), 3.05 (2H, m), 3.10-3.78 (4H, m), 3.92 (1H, m), 4.55 (1H, m).

Step 4) Preparation of trans-N- (t-butyloxycarbonyl) -3- (N-t-butyloxycarbonyl) amino-4- (N-t-butyloxycarbonyl) aminomethylpyrrolidine

694 mg of trans-N- (t-butyloxycarbonyl) -3- (Nt-butyloxycarbonyl) amino-4-azidomethylpyrrolidine is dissolved in 150 ml of methanol, where 10% Pd-C 200 mg was added and stirred overnight under 60 psi of hydrogen atmosphere. After filtration through celite, washed twice with 100 ml of methanol and twice with water, and then added 600 mg of di-t-butyloxycarbodicarbonate to the filtrate and reacted overnight. The solvent was removed, extracted with ethyl acetate, dried over anhydrous magnesium sulfate, and the solvent was removed. Then, silica gel column chromatography (eluate: ethyl acetate: hexane = 1: 3) gave 0.49 g of the target compound.

¹ H NMR (CDCl ₃ ): δ 1.37 (27H, s), 2.05 (1H, m), 2.93-3.90 (9H, m), 4.60 (1H, m)

Step 5) Preparation of trans-3-amino-4-aminomethylpyrrolidine trichloride

0.49 g of trans-N- (t-butyloxycarbonyl) -3- (Nt-butyloxycarbonyl) amino-4- (Nt-butyloxycarbonyl) aminomethylpyrrolidine is dissolved in 2 ml methanol, 15 ml of 9.8% methanol solution of hydrogen chloride were added dropwise. A precipitate formed after 7 hours of reaction was filtered to obtain 0.2 g of the target compound.

¹ H NMR (D ₂ O): δ 2.81 (1H, m), 3.06 (1H, q), 3.22 (1H, q), 3.34 (1H, q), 3.42 (1H, q), 3.76 (2H, m ), 3.90 (1 H, m).

Preparation Example 2: Preparation of cis-3-amino-4-aminomethylpyrrolidine trichloride

Step 1) Preparation of cis-N-benzyl-3-azido-4-azidomethylpyrrolidine

To 8.94 g of cis-N-benzyl-3-methanesulfonyloxy-4-methanesulfonyloxymethylpyrrolidine, 100 ml of dimethylformamide and 8 g of sodium azide were added, followed by stirring at 110 ° C. for 2 hours. The reaction mixture was cooled, extracted with water and ethyl acetate, and then concentrated under reduced pressure with magnesium sulfate, followed by silica gel column chromatography (eluent: hexane: ethyl acetate = 4: 1) to obtain 5 g of the target compound.

¹ H NMR (CDCl ₃ ): δ 2.33 (1H, q) 2.36-2.49 (1H.m) 2.60 (1H, q) 2.70 (1H, q) 2.95 (1H, q) 3.25 (1H, q), 3.46 ( 1H, q) 3.60 (2H, s) 3.98 (1H, m) 7.17-7.26 (5H, m).

Step 2) Preparation of cis-N-benzyl-3- (t-butyloxycarbonyl) amino-4- (t-butyloxycarbonyl) aminomethylpyrrolidine

Dissolve 2.76 g of cis-N-benzyl-3-azido-4-azidomethylpyrrolidine in 20 ml of tetrahydrofuran, add 6.5 g of triphenylphosphine, stir at room temperature for 15 hours, add 2 ml of water, and add another 8 hours. Stirred. After the solvent was removed, 5.6 g of di-tert-butyldicarbonate was added in methanol, followed by stirring for 5 hours. The solvent was removed and silica gel column chromatography (eluent: hexane: ethyl acetate = 1: 1) gave 2.43 g of the target compound.

¹ H NMR (CDCl ₃ ): δ 1.42 (18H, s) 2.39-2.52 (3H.m) 2.60 (2H.m) 2.63 (1H, m) 3.36 (1H, m) 3.57 (2H, q) 4.18 (1H , m) 4.97 (1H, br) 5.62 (1H, br) 7.23-7.32 (5H, m)

Step 3) Preparation of cis-N- (t-butyloxycarbonyl) -3- (t-butyloxycarbonyl) amino-4- (t-butyloxycarbonyl) aminomethylpyrrolidine

Dissolve 2.43 g of cis-N-benzyl-3- (Nt-butyloxycarbonyl) amino-4- (Nt-butyloxycarbonyl) aminomethylpyrrolidine in methanol, add 300 mg of Pd-C and hydrogenate for 12 hours Reacted. Pd-C was filtered through celite, and 2.5 g of di-t-butyldicarbonate was added to the filtrate and stirred for 5 hours. After removing the solvent, silica gel column chromatography (eluent: hexane: ethyl acetate = 2: 1) gave 1.65 g of the target compound.

¹ H NMR (CDCl ₃ ): δ 1.36 (27H, s) 2.20-2.40 (1H, m) 2.60-2.92 (2H, m) 3.15-3.49 (4H, m) 4.10-4.19 (1H, m) 4.83-5.05 (1H, br) 5.32-5.60 (1H, br)

Step 4) Preparation of cis-3-amino-4-aminomethylpyrrolidine trichloride

1.65 g of cis-N- (t-butyloxycarbonyl) -3- (t-butyloxycarbonyl) amino-4- (t-butyloxycarbonyl) aminomethylpyrrolidine was added to 50 ml of 9.8% hydrogen chloride methanol solution. It was added and stirred for 8 hours. The resulting white precipitate was filtered to give 733 mg of the target compound.

¹ H NMR (D ₂ O): δ 2.85-2.98 (1H, m) 3.08 (1H, q) 3.25-3.33 (2H, m) 3.50 (1H, q) 3.67-3.81 (2H, m) 4.19-4.25 ( 1H, m)

Preparation Example 3 Preparation of 3-Amino-4- (N-methyl) aminomethylpyrrolidine

Step 1) Preparation of N- (t-butyloxycarbonyl) -3- (N-t-butyloxycarbonyl) amino-4- (N-t-butyloxycarbonyl-N-methyl) aminomethylpyrrolidine

1.8 g of N- (t-butyloxycarbonyl) -3- (methanesulfonyl) oxymethyl-4- (Nt-butyloxycarbonyl) aminopyrrolidine, 150 ml of 40% methylamine aqueous solution and 100 ml of methanol After addition to the flask, the inlet of the flask was sealed. After reacting at 65 ° C. for 8 hours, the solvent was evaporated, water was added, extracted with ethyl acetate, and the solvent was removed. The residue was dissolved in 50 ml of methanol and reacted for 10 hours by adding 1.4 g of di-t-butyldicarbonate, and then the solvent was removed and silica gel column chromatography (eluate: ethyl acetate: hexane = 1: 3) gave the target compound 0.98. g was obtained.

¹ H NMR (CDCl ₃ ): δ 1.44 (27H, s), 2.3 (1H, m), 2.86 (3H, s), 3.05-3.90 (7H, m), 4.6 (1H, br d).

Step 2) Preparation of 3-Amino-4- (N-methyl) aminomethylpyrrolidine Hydrogen Trichloride

Dissolve 0.98 g of N- (t-butyloxycarbonyl) -3- (Nt-butyloxycarbonyl) amino-4- (Nt-butyloxycarbonyl-N-methyl) aminomethylpyrrolidine in 5 ml of methanol 40 ml of 9.8% methanol solution of hydrogen chloride was added thereto, and the mixture was reacted at room temperature for 7 hours, and then the resulting precipitate was filtered to obtain 0.4 g of the target compound.

¹ H NMR (D ₂ O): δ 2.63 (3H, s), 2.86 (1H, m), 3.19 (1H, q), 3.26 (1H, q), 3.36 (1H, q), 3.43 (1H, q), 3.77 (2H, m), 3.90 (1H, m)

Preparation Example 4 Preparation of 3-methylamino-4-aminomethylpyrrolidine

Step 1) Preparation of N-benzyl-3- (N-benzyl-N-t-butyloxycarbonyl) amino-4-hydroxymethylpyrrolidine

4 g of lithium aluminum hydride was added to 300 ml of purified tetrahydrofuran under a nitrogen atmosphere, and 35 g of N-benzyl-3-benzylamino-4-ethoxycarbonylpyrrolidine was slowly added to 200 ml of tetrahydrofuran while stirring. . After stirring overnight, it was heated to reflux for 2 hours. Water was added slowly to terminate the reaction, and then the solvent was removed, and 500 ml of methanol and 26 g of di-tert-butyldicarbonate were added thereto, followed by stirring at room temperature for 2 days. The reaction product was filtered, the solvent was removed, water and ethyl acetate were added, the organic layer was concentrated under reduced pressure and dried over magnesium sulfate, and then silica gel column chromatography (eluent: hexane: ethyl acetate = 2: 1) to obtain 29.5 g of the target compound.

¹ H NMR (CDCl ₃ ): δ 1.25 (9H, s) 2.34 (2H, m) 2.71 (1H, m) 2.98 (6H, m) 3.39-3.56 (4H, m) 4.00 (1H, m) 4.41 (2H m) 4.57 (2H, q) 7.15-7.28 (5H, m)

Step 2) Preparation of N-benzyl-3- (N-benzyl-N-methyl) amino-4-hydroxymethylpyrrolidine

0.2 g of lithium aluminum hydride was added to 20 ml of tetrahydrofuran under nitrogen, and 1.0 g of N-benzyl-3- (N-benzyl-Nt-butyloxycarbonyl) amino-4-hydroxymethylpyrrolidine was added with tetra Diluted in 20 ml of hydrofuran and added slowly. After stirring for 3 hours at room temperature, water was added slowly to terminate the reaction. After the reaction was filtered, the solvent was removed, and silica gel column chromatography (eluent: hexane: ethyl acetate = 1: 2) gave 753 mg of the target compound.

¹ H NMR (CDCl ₃ ): δ 2.15 (3H, s) 2.38 (1H, m) 2.55 (2H, m) 2.68 (1H, m) 2.86 (1H, m) 3.10 (1H, m) 3.50-3.65 (5H , m) 3.71 (1 H, m) 3.90 (1 H, br)

Step 3) Preparation of N-benzyl-3- (N-benzyl-N-methyl) amino-4-methanesulfonyloxymethylpyrrolidine

1.2 g of N-benzyl-3- (N-benzyl-N-methyl) amino-4-hydroxymethylpyrrolidine is dissolved in 15 ml of purified dichloromethane, where 0.65 ml of triethylamine and methanesulfur at 0 ° C. 0.36 ml of polyvinylchloride was added and stirred for 30 minutes. After adding water, the mixture was extracted twice with dichloromethane, dried over magnesium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography (eluate: hexane: ethyl acetate = 2: 1) to obtain 1.43 g of the target compound.

¹ H NMR (CDCl ₃ ): δ 2.16 (3H, s) 2.46 (1H, m) 2.58 (2H, m) 2.72 (2H, m) 2.91 (2H, s) 3.49 (2H, s) 3.57 (2H, q ) 4.12 (1H, m) 4.26 (1H, m)

Step 4) Preparation of N-benzyl-3- (N-benzyl-N-methyl) amino-4-azidomethylpyrrolidine

1.3 g of N-benzyl-3- (N-benzyl-N-methyl) amino-4-methanesulfonyloxymethylpyrrolidine was added to 20 ml of dimethylformamide, and 1.0 g of sodium azide was added thereto, followed by 100 DEG C. Heated and stirred for 5 hours. The reaction was cooled, water and ethyl acetate were added, extracted, concentrated under reduced pressure with magnesium sulfate, and then silica gel column chromatography (eluent: hexane: ethyl acetate = 4: 1) to obtain 1.0 g of the target compound.

¹ H NMR (CDCl ₃ ): δ 2.15 (3H, s) 2.36 (2H.m) 2.62 (2H, m) 2.74 (1H, m) 2.86 (1H, m) 3.28 (1H, m) 3.43 (1H, m ) 3.55 (4H, m)

Step 5) Preparation of N-t-butyloxycarbonyl-3- (N-t-butyloxycarbonyl-N-methyl) amino-4- (t-butyloxycarbonyl) aminomethylpyrrolidine

After dissolving 900 mg of N-benzyl-3- (N-benzyl-N-methyl) amino-4-azidomethylpyrrolidine in methanol, 600 mg of 10% Pd-C was added and hydrogenated for 10 hours. Pd-C was filtered through celite and 3 g of di-t-butyldicarbonate was added to the filtrate, followed by stirring for 8 hours. After removing the solvent, silica gel column chromatography (eluent: hexane: ethyl acetate = 2: 1) gave 680 mg of the target compound.

¹ H NMR (CDCl ₃ ): δ 1.40 (18H, s) 1.45 (9H, s) 2.33 (1H, m) 2.73 (3H, s) 2.94 (1H, m) 3.05 (1H, m) 3.23 (2H, m ) 3.54 (2H, m) 4.55 (1H, m)

Step 6) Preparation of 3-methylamino-4-aminomethylpyrrolidine trichloride

680 mg of Nt-butyloxycarbonyl-3- (Nt-butyloxycarbonyl-N-methyl) amino-4- (t-butyloxycarbonyl) aminomethylpyrrolidine was added to 15 ml of 9.8% hydrogen chloride solution and stirred overnight. After filtering the white precipitate produced to obtain 350mg of the target compound.

¹ H NMR (D ₂ O): δ 2.62 (3H, s) 2.87-2.93 (1H, m) 3.10 (1H, q) 3.28-3.38 (2H, m) 3.55 (1H, q) 3.71-3.82 (2H, m) 3.84-3.91 (1H, m)

Preparation Example 5 Preparation of 3- (N-3′-pyridinemethyl) amino-4-aminomethylpyrrolidine

Step 1) Preparation of N-t-butyloxycarbonyl-3- (N-3'-pyridinemethyl) amino-4-azidomethylpyrrolidine

234 mg of sodium hydride was added to 10 ml of dimethylformamide, and 1 g of Nt-butyloxycarbonyl-3- (Nt-butyloxycarbonyl) amino-4-azidomethylpyrrolidine was dissolved in 5 ml of dimethylformamide while stirring. Was added slowly and stirred for 1 h. 655 mg of 3-methanesulfonyloxymethylpyridine hydrochloride was added to the reaction solution and then heated at 40 ° C. for 4 hours. The reaction solution was cooled, extracted with water and ethyl acetate, and then concentrated under reduced pressure with magnesium sulfate, followed by silica gel column chromatography (eluent: ethyl acetate) to obtain the title compound (882 mg).

¹ H NMR (CDCl ₃ ): δ 1.38 (9H, s), 2.19 (1H, m), 3.08 (2H, m), 3.41 (3H, m), 3.75 (1H, m), 4.02 (1H, m) , 5.20 (2H, s), 7.23 (1H, m), 7.63 (1H, d), 8.52 (2H, m)

Step 2) Preparation of N-t-butyloxycarbonyl-3- (N-3'-pyridinemethyl) amino-4- (t-butyloxycarbonyl) aminomethylpyrrolidine

Nt-butyloxycarbonyl-3- (N-3'-pyridinemethyl) amino-4-azidomethylpyrrolidine was dissolved in 15 ml of 882 mg tetrahydrofuran, 871 mg of triphenylphosphine was added thereto, and the mixture was stirred at room temperature for 14 hours. . Was added to the reaction solution, and stirred for 8 hours. The solvent was removed, the residue was dissolved in methanol, 723 g of di-t-butyldicarbonate was added thereto, the mixture was stirred for 5 hours, the solvent was removed, and the residue was purified by silica gel column chromatography (eluent: ethyl acetate) to obtain the title compound. 516 mg of compound was obtained.

¹ H NMR (CDCl ₃ ): δ 1.32 (18H, s), 2.15 (1H, m), 3.03 (3H, m), 3.28 (1H, m), 3.52 (1H, m), 3.72 (1H, q) , 3.93 (1H, s), 5.03 (2H, s), 5.43 (1H, d), 7.20 (1H, m), 7.63 (1H, m), 8.49 (2H, m)

Step 3) Preparation of 3- (N-3'-pyridinemethyl) amino-4-aminomethylpyrrolidine tetrachloride

To 483 mg of trans-Nt-butyloxycarbonyl-3- (N-3'-pyridinemethyl) amino-4- (t-butyloxycarbonyl) aminomethylpyrrolidine, 15 ml of 9.8% hydrogen chloride methanol solution was added and stirred overnight. After filtration, the produced white precipitate was filtered to obtain 126 mg of the target compound.

¹ H NMR (D ₂ O): δ 2.51-2.59 (1H, m), 2.99-3.09 (1H, m), 3.13-3.18 (2H, m), 3.55 (1H, q), 3.65 (1H, q) , 4.03 (1H, q), 5.20 (1H, q), 7.93 (1H, q), 8.93 (1H, d), 8.61 (1H, d), 8.67 (1H, s)

Preparation Example 6 Preparation of 3-benzylamino-4-aminomethylpyrrolidine

3.5 g of N-benzyl-3- (N-benzyl-Nt-butyloxycarbonyl) amino-4-hydroxymethylpyrrolidine was treated in the same manner as in steps 3 to 6 of Preparation Example 4 to obtain 650 mg of the target compound. .

¹ H NMR (D ₂ O): δ 2.91-2.99 (1H, m), 3.12 (1H, q), 3.31-3.48 (2H, m), 3.61 (1H, q), 3.71-3.82 (4H, m) , 3.84-3.91 (1H, m), 7.23-7.32 (5H, m)

Preparation Example 7 Preparation of 3-methylamino-4-methylaminomethylpyrrolidine

Step 1) Preparation of N-benzyl-3-azido-4-azidomethylpyrrolidine

8.94 g of N-benzyl-3-methanesulfonyloxy-4-methanesulfonyloxymethylpyrrolidine and 8 g of sodium azide were added to 100 ml of dimethylformamide and heated and stirred at 110 ° C. for 2 hours. After the reaction solution was cooled, water and ethyl acetate were added for extraction, dried over magnesium sulfate, concentrated under reduced pressure, and silica gel column chromatography (eluent: hexane: ethyl acetate = 4: 1) to obtain 5 g of the target compound.

¹ H NMR (CDCl ₃ ): δ 2.33 (1H, q) 2.36-2.49 (1H.m) 2.60 (1H, q) 2.70 (1H, q) 2.95 (1H, q) 3.25 (1H, q) 3.46 (1H , q) 3.60 (2H, s) 3.98 (1H, m) 7.17-7.26 (5H, m)

Step 2) Preparation of N-benzyl-3- (N-t-butyloxycarbonyl) amino-4- (N-t-butyloxycarbonyl) aminomethylpyrrolidine

2.76 g of N-benzyl-3-azido-4-azidomethylpyrrolidine was dissolved in 20 ml of tetrahydrofuran, and 6.5 g of triphenylphosphine was added thereto, followed by stirring at room temperature overnight. Subsequently, 1 ml of water was added to the reaction solution, followed by stirring for 8 hours. After the solvent was removed, the residue was dissolved in 100 ml of methanol, and 5.6 g of di-tert-butyldicarbonate was added and stirred for 5 hours. The solvent was removed, the residue was extracted with 100 ml of ethyl acetate, the extract was dried over magnesium sulfate, concentrated under reduced pressure and silica gel column chromatography (eluate: hexane: ethyl acetate = 1: 1) to obtain 2.43 g of the target compound.

¹ H NMR (CDCl ₃ ): δ 1.42 (18H, s) 2.39-2.52 (3H.m) 2.60 (2H.m) 2.63 (1H, m) 3.36 (1H, m) 3.57 (2H, q) 4.18 (1H , m) 4.97 (1H, br) 5.62 (1H, br) 7.23-7.32 (5H, m)

Step 3) Preparation of N-benzyl-3- (N-methyl-N-t-butyloxycarbonyl) amino-4- (N-methyl-N-t-butyloxycarbonyl) aminomethylpyrrolidine

2.43 g of N-benzyl-3- (Nt-butyloxycarbonyl) amino-4- (Nt-butyloxycarbonyl) aminomethylpyrrolidine is dissolved in 30 ml of tetrahydrofuran, and 300 mg of lithium aluminum hydride is added thereto. The reaction was carried out for 12 hours. 1 ml of water, 30 ml of methanol and 2.5 g of di-t-butyldicarbonate were added to the reaction solution, and the mixture was stirred for 5 hours. After removing the solvent, silica gel column chromatography (eluent: hexane: ethyl acetate = 2: 1) gave 1.65 g of the target compound.

¹ H NMR (CDCl ₃ ): δ 1.36 (27H, s) 2.20-2.40 (1H, m) 2.60-2.92 (2H, m) 3.15-3.49 (4H, m) 4.10-4.19 (1H, m) 4.83-5.05 (1H, br) 5.32-5.60 (1H, br)

Step 4) of N- (t-butyloxycarbonyl) -3- (N-methyl-Nt-butyloxycarbonyl) amino-4- (N-methyl-Nt-butyloxycarbonyl) aminomethylpyrrolidine Produce

2.63 g of N-benzyl-3- (N-methyl-Nt-butyloxycarbonyl) amino-4- (N-methyl-Nt-butyloxycarbonyl) aminomethylpyrrolidine is dissolved in methanol, where Pd 300 mg of -C was added and hydrogenated for 10 hours. Pd-C was filtered through celite, and 2.4 g of di-t-butyldicarbonate was added to the filtrate, followed by stirring for 12 hours. After removing the solvent, silica gel column chromatography (eluate: hexane: ethyl acetate = 2: 1) gave 1.71 g of the target compound.

Step 5) Preparation of 3-methylamino-4-methylaminomethylpyrrolidine trichloride

1.45 g of N- (t-butyloxycarbonyl) -3- (N-methyl-Nt-butyloxycarbonyl) amino-4- (N-methyl-Nt-butyloxycarbonyl) aminomethylpyrrolidine 50 ml of% hydrogen chloride solution was added and stirred for 8 hours. The resulting white precipitate was filtered to give 771 mg of the target compound.

¹ H NMR (D ₂ O): δ 2.85-2.98 (7H, m) 3.08 (1H, q) 3.25-3.33 (2H, m) 3.50 (1H, q) 3.67-3.81 (2H, m) 4.19-4.25 ( 1H, m)

Preparation Example 8 Preparation of 3-Amino-4- (N-2-fluoroethyl) aminomethylpyrrolidine Trichloride

Step 1) Nt-Butyloxycarbonyl-3- (N-benzyl-Nt-butyloxycarbonyl) amino-4- (N- (2-fluoroethyl) -Nt-butyloxycarbonyl) aminomethylpyrrolidine Manufacture

280 mg of sodium hydride was added to 10 ml of dimethylformamide and the mixture was stirred with Nt-butyloxycarbonyl-3- (N-benzyl-Nt-butyloxycarbonyl) amino-4- (t-butyloxycarbonyl) amino A solution of 1.2 g of tilpyrrolidine dissolved in 20 ml of dimethylformamide was slowly added and stirred for 2 hours. 620 mg of 2-fluoromethanesulfonyloxyethane was added to the reaction solution and heated at 40 ° C. for 6 hours. The reaction solution was cooled, extracted with ethyl acetate, dried over magnesium sulfate, concentrated under reduced pressure, and silica gel column chromatography (eluent: hexane: ethyl acetate = 2: 1) to obtain 1.22 g of the target compound.

¹ H NMR (CDCl ₃ ): δ 1.38 (9H, s), 1.39 (18H, s), 2.21 (1H, m), 2.33 (2H, m), 3.15 (2H, m), 3.42 (5H, m) , 3.77 (1H, m), 4.12 (1H, m)

Step 2) Preparation of N-t-butyloxycarbonyl-3- (N-t-butyloxycarbonyl) amino-4- (N- (2-fluoroethyl) -N-t-butyloxycarbonyl) aminomethylpyrrolidine

2.50 g of Nt-butyloxycarbonyl-3- (N-benzyl-Nt-butyloxycarbonyl) amino-4- (N- (2-fluoroethyl) -Nt-butyloxycarbonyl) aminomethylpyrrolidine After dissolving in methanol, 600% of 10% Pd-C was added and hydrogenated for 12 hours. After the catalyst was filtered off, 2.5 g of di-t-butyldicarbonate was added to the filtrate and stirred for 5 hours. After removing the solvent, silica gel column chromatography (eluent: hexane: ethyl acetate = 2: 1) gave 2.25 g of the target compound.

¹ H NMR (CDCl ₃ ): δ 1.36 (27H, s), 2.20-2.30 (3H, m), 2.60-2.92 (2H, m), 3.15-3.49 (6H, m), 4.15 (1H, m)

Step 3) Preparation of 3-amino-4- (N- (2-fluoroethyl)) aminomethylpyrrolidine trihydrochloride

Nt-butyloxycarbonyl-3- (Nt-butyloxycarbonyl) amino-4- (N- (2-fluoroethyl) -Nt-butyloxycarbonyl) aminomethylpyrrolidine 410mg 9.8% hydrogen chloride methanol solution After addition to 15ml and stirred overnight, the resulting white precipitate was filtered to give 167mg of the target compound.

¹ H NMR (D ₂ O): δ 2.51-2.59 (3H, m), 2.99-3.09 (3H, m), 3.13-3.18 (3H, m), 3.55 (2H, q), 4.05 (1H, q)

Preparation Example 9 Preparation of 3- (N-2-fluoroethyl) amino-4-aminomethylpyrrolidine Trichloride

Step 1) Preparation of N-t-butyloxycarbonyl-3- (N- (2-fluoroethyl) -N-t-butyloxycarbonyl) amino-4-azidomethylpyrrolidine

234 mg of sodium hydride was added to 10 ml of dimethylformamide, and 1 g of Nt-butyloxycarbonyl-3- (Nt-butyloxycarbonyl) amino-4-azidomethylpyrrolidine was dissolved in 5 ml of dimethylformamide while stirring. Was added slowly and stirred for 1 h. 426 mg of 2-fluoromethanesulfonyloxyethane was added to the reaction solution and then heated at 40 ° C. for 4 hours. The reaction solution was cooled, added with water, extracted with ethyl acetate, dried over magnesium sulfate, concentrated under reduced pressure, and silica gel column chromatography (hexane: ethyl acetate = 2: 1) to give 882 mg of the target compound.

¹ H NMR (CDCl ₃ ): δ 1.38 (9H, s), 1.40 (9H, s), 2.19 (1H, m), 2.30 (2H, m), 3.08 (2H, m), 3.41 (5H, m) , 3.75 (1H, m), 4.02 (1H, m)

Step 2) Preparation of N-t-butyloxycarbonyl-3- (N- (2-fluoroethyl) -N-t-butyloxycarbonyl) amino-4- (N-t-butyloxycarbonyl) aminomethylpyrrolidine

811 mg of Nt-butyloxycarbonyl-3- (N- (2-fluoroethyl) -Nt-butyloxycarbonyl) amino-4-azidomethylpyrrolidine was dissolved in 15 ml of tetrahydrofuran, and triphenyl was added thereto. 850 mg of phosphine was added and stirred at room temperature for 10 hours. Water was added to the reaction solution and stirred for 8 hours. After the solvent was removed, the residue was dissolved in methanol, and 1.2 g of di-t-butyldicarbonate was added thereto and stirred for 5 hours. After the solvent was removed, silica gel column chromatography (eluent: hexane: ethyl acetate = 2: 1) gave 495 mg of the target compound.

¹ H NMR (CDCl ₃ ): δ 1.32 (18H, s), 2.15 (1H, m), 2.27 (2H, m), 3.03 (3H, m), 3.28 (3H, m), 3.52 (1H, m) , 3.72 (1H, q), 3.93 (1H, s)

Step 3) Preparation of 3- (N- (2-fluoroethyl)) amino-4-aminomethylpyrrolidine trihydrochloride

385 mg of Nt-butyloxycarbonyl-3- (N- (2-fluoroethyl)) amino-4- (t-butyloxycarbonyl) aminomethylpyrrolidine was added to 15 ml of 9.8% methanol solution of hydrogen chloride and stirred overnight. The produced white precipitate was filtered to give 147 mg of the target compound.

¹ H NMR (D ₂ O): δ 2.51-2.59 (3H, m), 2.99-3.09 (3H, m), 3.13-3.18 (3H, m), 3.55 (1H, q), 3.65 (1H, q) , 4.03 (1H, q)

Preparation Example 10 Preparation of 3-methoxyamino-4-aminomethylpyrrolidine Trichloride

Step 1) Preparation of N-benzyl-3-methanesulfonyloxy-4-azidomethylpyrrolidine

20 g of N-benzyl-3-methanesulfonyloxy-4-methanesulfonyloxymethylpyrrolidine and 5.43 g of sodium azide were added to 300 ml of dimethylformamide and then stirred at 35 ° C. for 10 hours. After the reaction solution was cooled, water and ethyl acetate were added for extraction. The organic phase was dried over magnesium sulfate, concentrated under reduced pressure, and then subjected to silica gel column chromatography (eluent: hexane: ethyl acetate = 1: 1) to obtain 17 g of the target compound.

¹ H NMR (CDCl ₃ ): δ 2.23 (1H.m) 2.52-2.57 (1H, m) 2.85 (2H, m) 3.44 (2H, m) 3.58 (2H, q) 4.84 (1H.m) 7.22-7.35 (5H, m)

Step 2) Preparation of N-benzyl-3- (N-methoxy-N-t-butyloxycarbonyl) amino-4-azidomethylpyrrolidine

0.748 g of sodium hydride was added to 20 ml of dimethylformamide, and then a solution in which 2.75 g of methoxy- (t-butyloxycarbonyl) amine was dissolved in 5 ml of dimethylformamide was slowly added while stirring. After stirring for 2 hours, a solution of 3.87 g of N-benzyl-3-methanesulfonyloxy-4-azidomethylpyrrolidine dissolved in 10 ml of dimethylformamide was slowly added, followed by stirring at 90 ° C. for 16 hours. After the reaction solution was cooled, water and ethyl acetate were added for extraction. The ethyl acetate layer was concentrated under reduced pressure with magnesium sulfate, and the residue was purified by silica gel column chromatography (eluent: hexane: ethyl acetate = 2: 1) to obtain 3.22 g of the target compound.

¹ H NMR (CDCl ₃ ): δ 1.50 (9H, s) 2.23 (1H, t) 2.53-2.61 (2H, m) 3.00-3.14 (2H, m) 3.40 (2H, d) 3.65 (3H, s) 3.62 -3.73 (2H, m) 4.69 (1H, q) 7.23-7.36 (5H, m)

Step 3) Preparation of N-benzyl-3- (N-methoxy-N-t-butyloxycarbonyl) amino-4- (N-t-butyloxycarbonyl) aminomethylpyrrolidine

3.22 g of N-benzyl-3- (N-methoxy-Nt-butyloxycarbonyl) amino-4-azidomethylpyrrolidine was dissolved in 30 ml of tetrahydrofuran and 2.8 g of triphenylphosphine was added to the solution for 12 hours. After stirring at room temperature, 1 ml of water was added and stirring was continued for 8 hours. The solvent was removed, the residue was dissolved in methanol, and 2.34 g of di-tert-butyldicarbonate was added thereto and stirred for 5 hours. The solvent was removed, and silica gel column chromatography (eluent: hexane: ethyl acetate = 1: 1) gave 2.18 g of the target compound.

¹ H NMR (CDCl ₃ ): δ 1.34 (9H, s) 1.40 (9H, s) 2.18 (1H, t) 2.37-2.45 (1H, m) 2.55 (1H, m) 2.88 (1H, q) 3.05 (2H , m) 3.23 (1H, m) 3.56 (2H, q) 3.61 (3H, s) 4.60 (1H, q) 4.90 (1H, br) 7.16-7.25 (5H, m)

Step 4) Preparation of N-trichloroethyloxycarbonyl-3- (N-methoxy-N-t-butyloxycarbonyl) amino-4- (N-t-butyloxycarbonyl) aminomethylpyrrolidine

625 mg of N-benzyl-3- (N-methoxy-Nt-butyloxycarbonyl) amino-4- (Nt-butyloxycarbonyl) aminomethylpyrrolidine is dissolved in 8 ml of dichloromethane, and 2,2 0.4 ml of 2-trichloroethylchloroformate and 62.3 mg of sodium bicarbonate were added thereto, and the resulting mixture was heated to reflux for 5 hours. Water was added to separate the organic layer, and the organic layer was dried over magnesium sulfate, concentrated under reduced pressure and silica gel column chromatography (eluate: hexane: ethyl acetate = 2: 1) to obtain 670 mg of the target compound.

¹ H NMR (CDCl ₃ ): δ 1.36 (9H, s) 1.42 (9H, s) 2.53 (1H, m) 2.88-2.93 (1H, m) 3.30 (1H, q) 3.42 (2H, m) 3.64 (3H s) 3.54-3.83 (2H, m) 4.62 (2H, m) 4.76 (1H, q) 5.07 (1H, br)

Step 5) Preparation of N-t-butyloxycarbonyl-3- (N-methoxy-N-t-butyloxycarbonyl) amino-4- (N-t-butyloxycarbonyl) aminomethylpyrrolidine

670 mg of N-trichloroethyloxycarbonyl-3- (N-methoxy-Nt-butyloxycarbonyl) amino-4- (Nt-butyloxycarbonyl) aminomethylpyrrolidine was dissolved in 10 ml of tetrahydrofuran Then, 1.26 g of activated zinc was added thereto, followed by heating to reflux for 5 hours. The reaction was cooled, filtered and the solvent removed, dissolved in 30 ml of methanol, 3.5 ml of triethylamine and 400 mg of di-tert-butyldicarbonate were added and stirred at room temperature for 3 hours. After removing the solvent, silica gel column chromatography (eluent: dichloromethane: ethyl acetate = 4: 1) gave 379 mg of the target compound.

¹ H NMR (CDCl ₃ ): δ 1.39 (9H, s) 1.43 (9H, s) 1.49 (9H, s) 2.50 (1H, m) 2.83-3.02 (1H, m) 3.13 (1H, q) 3.42 (2H , m) 3.67 (3H, s) 3.56-3.72 (2H, m) 4.58 (1H, m) 5.08 (1H, br)

Step 6) Preparation of 3-methoxyamino-4-aminomethylpyrrolidine trichloride

To 379 mg of Nt-butyloxycarbonyl-3- (N-methoxy-Nt-butyloxycarbonyl) amino-4- (Nt-butyloxycarbonyl) aminomethylpyrrolidine, add 7 ml of 9.8% hydrogen chloride methanol solution to 12 Stir for hours. The produced white precipitate was filtered to give 170 mg of the target compound.

¹ H NMR (D ₂ O): δ 2.62-2.69 (2H, m) 3.03-3.18 (3H, m) 3.36 (3H, s) 3.34-3.47 (2H, m) 3.72-3.80 (1H, m)

Preparation Example 11 Preparation of 3-Amino-4- (N-methoxy) aminomethylpyrrolidine Trichloride

Step 1) Preparation of N-t-butyloxycarbonyl-3- (N-t-butyloxycarbonyl) amino-4- (N-methoxy-N-t-butyloxycarbonyl) aminomethylpyrrolidine

242 mg sodium hydride and 893 mg t-butyloxycarbonylmethoxyamine are added to 30 ml of dimethylformamide and stirred until gas evolution stops. To this was added 1.596 g of N-t-butyloxycarbonyl-3- (N-t-butyloxycarbonyl) amino-4-methanesulfonyloxymethylpyrrolidine and heated at 90 ° C overnight. The reaction was cooled, water was added and extracted three times with ethyl acetate. The organic layer was dried over magnesium sulfate, concentrated under reduced pressure, and then silica gel column chromatography (eluate: hexane: ethyl acetate = 2: 1) to obtain 1.05 g of the target compound.

¹ H NMR (CDCl ₃ ): δ 1.40 (18H, s) 1.46 (9H, s) 2.30 (1H, m) 3.03 (2H, m) 3.35 (1H, m) 3.58 (1H, m) 3.64 (3H, s ) 3.71 (1H, m) 3.89 (1H, m) 4.67 (1H, br)

Step 2) Preparation of 3-Amino-4-methoxyaminomethylpyrrolidine Trichloride

To 900 mg of Nt-butyloxycarbonyl-3- (t-butyloxycarbonyl) amino-4-methoxy- (t-butyloxycarbonyl) aminomethylpyrrolidine, add 15 ml of 9.8% methanol solution of hydrogen chloride and stir overnight. After filtration, the white precipitate was filtered to obtain 302 mg of the target compound.

¹ H NMR (D ₂ O): δ 2.87 (1H, m) 3.23 (1H, m) 3.39-3.42 (2H, m) 3.62 (1H, m) 3.74 (3H, s) 3.72-3.78 (2H, m) 3.89 (1H, m)

Example 1 1-cyclopropyl-6,8-difluoro-7-[(3-N-benzylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-di Preparation of Hydroquinoline-3-carboxylic Acid

1-cyclopropyl-6,7,8-trifluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid 280 mg, 3-N-benzylamino-4-aminomethylpyrrolidine trichloride 225 mg And 200 mg of diazabicyclo [5.4.0] undec-7-ene was added to 5 ml of acetonitrile, refluxed for 6 hours, cooled, filtered, and dried to obtain 354 mg of a pale yellow target compound.

Elemental Analysis (C ₂₅ H ₂₆ N ₄ O ₃ F ₂ )

Measured value (%); C: 64.11 H: 5.58 N: 12.01

Theoretical value (%); C: 64.10 H: 5.56 N: 11.97

Example 2: 1-cyclopropyl-6-fluoro-8-chloro-7-[(3-N-benzylamino-4-aminomethylpyrrodine) -1-yl] -4-oxo-1,4-di Preparation of Hydroquinoline-3-carboxylic Acid

298 mg of 1-cyclopropyl-6,7-difluoro-8-chloro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 3-N-benzylamino-4-aminomethylpyrrolidine 3 225 mg of hydrochloride were treated in the same manner as in Example 1 to obtain 376 mg of the target compound as a white solid.

Elemental Analysis (C ₂₅ H ₂₆ N ₄ O ₃ FCl)

Measured value (%); C: 61.21 H: 5.41 N: 11.60

Theoretical value (%); C: 61.19 H: 5.37 N: 11.56

Example 3: 1-cyclopropyl-6-fluoro-8-methoxy-7-[(3-N-benzylamino-4-aminomethylpyrrodine) -1-yl] -4-oxo-1,4- Preparation of Dihydroquinoline-3-carboxylic Acid

293 mg of 1-cyclopropyl-6,7-difluoro-8-methoxy-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 3-N-benzylamino-4-aminomethylpyrrolidine 225 mg of trichloride was treated in the same manner as in Example 1 to obtain 382 mg of the target compound.

Elemental Analysis (C ₂₆ H ₂₉ N ₄ O ₄ F)

Measured value (%); C: 65.06 H: 5.99 N: 11.77

Theoretical value (%); C: 65.00 H: 6.04 N: 11.67

Example 4: 1-cyclopropyl-5-amino-6,8-difluoro-7-[(3-N-benzylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1 Preparation of 4-, dihydroquinoline-3-carboxylic acid

295 mg of 1-cyclopropyl-5-amino-6,7,8-trifluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 3-N-benzylamino-4-aminomethylpyrroli 225 mg of Dean trihydrochloride was treated in the same manner as in Example 1 to obtain 396 mg of the yellow target compound.

Elemental Analysis (C ₂₅ H ₂₇ N ₅ O ₃ F ₂ )

Measured value (%); C: 62.17 H: 5.56 N: 14.41

Theoretical value (%); C: 62.11 H: 5.59 N: 14.49

Example 5 1-cyclopropyl-6-fluoro-7-[(3-N-benzylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline- Preparation of 3-carboxylic Acid

263 mg of 1-cyclopropyl-6,7-difluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 225 mg of 3-N-benzylamino-4-aminomethylpyrrolidine trichloride 361 mg of the target compound as a white solid was obtained in the same manner as in Example 1.

Elemental Analysis (C ₂₅ H ₂₇ N ₄ O ₃ F)

Measured value (%); C: 67.66 H: 5.94 N: 12.36

Theoretical value (%); C: 66.67 H: 6.00 N: 12.44

Example 6: 9-Fluoro-2,3-dihydro-3- (S) -methyl-10-[(3-N-benzylamino-4-aminomethylpyrrolidin) -1-yl] -7- Preparation of oxo-7H-pyrido [1.2.3-dec] -1,4-benzooxazine-6-carboxylic acid

9,10-Fluoro-2,3-dihydro-3- (S) -methyl-7-oxo-7H-pyrido [1.2.3-dec] -1,4-benzooxazine-6-carboxylic acid 287 mg and 225 mg of 3-N-benzylamino-4-aminomethylpyrrolidine trichloride were treated in the same manner as in Example 1 to obtain 364 mg of the target compound.

Elemental Analysis (C ₂₅ H ₂₇ N ₄ O ₄ F)

Measured value (%); C: 64.47 H: 5.91 N: 12.19

Theoretical value (%); C: 64.38 H: 5.79 N: 12.02

Example 7: 1-cyclopropyl-6-fluoro-7-[(3-N-benzylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydro-1 Preparation of 8, naphthyridine-3-carboxylic acid

264 mg of 1-cyclopropyl-6,7-difluoro-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid and 3-N-benzylamino-4-aminomethylpyrroli 225 mg of Dean trichloride was treated in the same manner as in Example to obtain 383 mg of the target compound.

Elemental Analysis (C ₂₄ H ₂₆ N ₅ O ₃ F)

Found (%); C: 63.85 H: 5.71 N: 15.71

Theoretical value (%); C: 63.86 H: 5.76 N: 15.52

Example 8 1- (2,4-Difluorophenyl) -6-fluoro-7-[(3-N-benzylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1 Preparation of, 4-dihydro-1,8-naphthyridine-3-carboxylic acid

334 mg of 1- (2,4-difluorophenyl) -6,7-difluoro-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid and 3-N-benzylamino 225 mg of 4-aminomethylpyrrolidine trichloride was treated in the same manner as in Example 1 to obtain 413 mg of the target compound.

Elemental Analysis (C ₂₇ H ₂₄ N ₅ O ₃ F ₃ )

Measured value (%); C: 62.03 H: 4.51 N: 13.45

Theoretical value (%); C: 61.95 H: 4.59 N: 13.38

Example 9 1-cyclopropyl-6,8-difluoro-7-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline- Preparation of 3-carboxylic Acid

Examples of 280 mg of 1-cyclopropyl-6,7,8-trifluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 126 mg of 3-amino-4-aminomethylpyrrolidine trichloride Treatment in the same manner as 1 afforded 320 mg of the target compound.

Elemental Analysis (C ₁₈ H ₂₀ N ₄ O ₃ F ₂ )

Measured value (%); C: 56.99 H: 5.21 N: 14.79

Theoretical value (%); C: 57.14 H: 5.29 N: 14.81

Example 10 1-cyclopropyl-6-fluoro-8-chloro-7-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline Preparation of 3-carboxylic acid

297 mg of 1-cyclopropyl-6,7-difluoro-8-chloro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 126 mg of 3-amino-4-aminomethylpyrrolidine trichloride 391 mg of the target compound as a white solid was obtained by the same method as in Example 1.

Elemental Analysis (C ₁₈ H ₂₀ N ₄ O ₃ FCl)

Measured value (%); C: 54.71 H: 4.98 N: 14.31

Theoretical value (%); C: 54.75 H: 5.07 N: 14.20

Example 11 1-cyclopropyl-6-fluoro-8-methoxy-7-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydro Preparation of Quinoline-3-carboxylic Acid

293 mg of 1-cyclopropyl-6,7-difluoro-8-chloro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 126 mg of 3-amino-4-aminomethylpyrrolidine trichloride The same procedure as in Example 1 was carried out to obtain 390 mg of the target compound as a white solid.

Elemental Analysis (C ₁₉ H ₂₃ N ₄ O ₄ F)

Measured value (%); C: 58.37 H: 5.81 N: 14.38

Theoretical value (%); C: 58.46 H: 5.90 N: 14.36

Example 12 1-cyclopropyl-5-amino-6,8-difluoro-7-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4- Preparation of Dihydroquinoline-3-carboxylic Acid

295 mg of 1-cyclopropyl-5-amino-6,7,8-trifluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 3-amino-4-aminomethylpyrrolidine trichloride 126 mg was treated in the same manner as in Example 1 to obtain 410 mg of the target compound.

Elemental Analysis (C ₁₈ H ₂₁ N ₅ O ₃ F ₂ )

Measured value (%); C: 54.89 H: 5.46 N: 17.88

Theoretical value (%); C: 54.96 H: 5.34 N: 17.81

Example 13: 9-Fluoro-2,3-dihydro-3- (S) -methyl-10-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -7-oxo-7H Preparation of pyrido [1.2.3-dec] -1,4-benzoxazine-6-carboxylic acid

9,10-Fluoro-2,3-dihydro-3- (S) -methyl-7-oxo-7H-pyrido [1.2.3-dec] -1,4-benzooxazine-6-carboxylic acid 287 mg and 126 mg of 3-amino-4-aminomethylpyrrolidine trichloride were treated in the same manner as in Example 1 to obtain 364 mg of the target compound.

Elemental Analysis (C ₁₈ H ₂₁ N ₄ O ₄ F)

Measured value (%); C: 57.54 H: 5.61 N: 14.80

Theoretical value (%); C: 57.45 H: 5.59 N: 14.89

Example 14 1-cyclopropyl-6-fluoro-7-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydro-1,8- Preparation of Naphthyridine-3-carboxylic Acid

264 mg of 1-cyclopropyl-6,7-fluoro-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid and 126 mg of 3-amino-4-aminomethylpyrrolidine trichloride Was treated in the same manner as in Example 1 to obtain 382 mg of the target compound.

Elemental Analysis (C ₁₇ H ₂₀ N ₅ O ₃ F)

Found (%); C: 56.39 H: 5.51 N: 19.31

Theoretical value (%); C: 56.50 H: 5.54 N: 19.39

Example 15 1-cyclopropyl-6-fluoro-7-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline-3-carr Preparation of Acids

295 mg of 1-cyclopropyl-6,7-difluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 126 mg of 3-amino-4-aminomethylpyrrolidine trichloride were compared with Example 1 The same procedure was followed to yield 327 mg of the target compound.

Elemental Analysis (C ₁₈ H ₂₁ N ₄ O ₃ F)

Measured value (%); C: 60.08 H: 5.81 N: 15.41

Theoretical value (%); C: 60.00 H: 5.83 N: 15.56

Example 16 1-cyclopropyl-6,8-difluoro-7-[(3-N-methylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-di Preparation of Hydroquinoline-3-carboxylic Acid

295 mg of 1-cyclopropyl-6,7,8-trifluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 141 mg of 3-N-methylamino-4-aminomethylpyrrolidine trichloride Was treated in the same manner as in Example 1 to obtain 327 mg of the target compound.

Elemental Analysis (C ₁₉ H ₂₂ N ₄ O ₃ F ₂ )

Measured value (%); C: 58.18 H: 5.51 N: 14.41

Theoretical value (%); C: 58.16 H: 5.61 N: 14.29

Example 17 1-cyclopropyl-6-fluoro-8-chloro-7-[(3-N-methylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4- Preparation of Dihydroquinoline-3-carboxylic Acid

298 mg of 1-cyclopropyl-6,7-difluoro-8-chloro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 3-N-methylamino-4-aminomethylpyrrolidine 3 141 mg of hydrochloride were treated in the same manner as in Example 1 to obtain 376 mg of the target compound as a white solid.

Elemental Analysis (C ₁₉ H ₂₂ N ₄ O ₃ FCl)

Measured value (%); C: 55.81 H: 5.43 N: 13.68

Theoretical value (%); C: 55.81 H: 5.39 N: 13.71

Example 18 1-cyclopropyl-6-fluoro-8-methoxy-7-[(3-N-methylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4 Preparation of Dihydroquinoline-3-carboxylic Acid

298 mg of 1-cyclopropyl-6,7-difluoro-8-methoxy-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 3-N-methylamino-4-aminomethylpyrrolidine 141 mg of trihydrochloride was treated in the same manner as in Example 1 to obtain 376 mg of the target compound as a white solid.

Elemental Analysis (C ₂₀ H ₂₅ N ₄ O ₄ F)

Measured value (%); C: 59.41 H: 6.23 N: 13.78

Theoretical value (%); C: 59.40 H: 6.19 N: 13.86

Example 19 1-cyclopropyl-5-amino-6,8-difluoro-7-[(3-N-methylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1 Preparation of 4-, dihydroquinoline-3-carboxylic acid

295 mg of 1-cyclopropyl-5-amino-6,7,8-trifluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 3-N-methylamino-4-aminomethylpyrroli 141 mg of Dean trihydrochloride were treated in the same manner as in Example 1 to obtain 370 mg of the target compound.

Elemental Analysis (C ₁₉ H ₂₃ N ₅ O ₃ F ₂ )

Measured value (%); C: 56.11 H: 5.56 N: 17.18

Theoretical value (%); C: 56.02 H: 5.65 N: 17.20

Example 20 9-Fluoro-2,3-dihydro-3- (S) -methyl-10-[(3-N-methylamino-4-aminomethylpyrrolidin) -1-yl] -7- Preparation of oxo-7H-pyrido [1.2.3-dec] -1,4-benzooxazine-6-carboxylic acid

9,10-Fluoro-2,3-dihydro-3- (S) -methyl-7-oxo-7H-pyrido [1.2.3-dec] -1,4-benzooxazine-6-carboxylic acid 287 mg and 141 mg of 3-N-methylamino-4-aminomethylpyrrolidine trichloride were treated in the same manner as in Example 1 to obtain 354 mg of the target compound.

Elemental Analysis (C ₁₉ H ₂₃ N ₄ O ₄ F)

Measured value (%); C: 58.47 H: 5.91 N: 14.40

Theoretical value (%); C: 58.46 H: 5.90 N: 14.36

Example 21 1-cyclopropyl-6,8-difluoro-7-[(3-amino-4-N-methylaminomethylpyrrolidin) -1-yl] -4-oxo-1,4-di Preparation of Hydroquinoline-3-carboxylic Acid

295 mg of 1-cyclopropyl-6,7,8-trifluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 141 mg of 3-amino-4-N-methylaminomethylpyrrolidine trichloride Was treated in the same manner as in Example 1 to obtain 377 mg of the target compound.

Elemental Analysis (C ₁₉ H ₂₂ N ₄ O ₃ F ₂ )

Found (%); C: 55.28 H: 5.51 N: 14.41

Theoretical value (%); C: 58.16 H: 5.61 N: 14.29

Example 22 1-cyclopropyl-6-fluoro-8-methoxy-7-[(3-amino-4-N-methylaminomethylpyrrolidin) -1-yl] -4-oxo-1,4 Preparation of Dihydroquinoline-3-carboxylic Acid

295 mg of 1-cyclopropyl-6,7-difluoro-8-methoxy-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 3-amino-4-N-methylaminomethylpyrrolidine 141 mg of trichloride was treated in the same manner as in Example 1 to obtain 397 mg of the target compound.

Elemental Analysis (C ₂₀ H ₂₅ N ₄ O ₃ F)

Measured value (%); C: 61.78 H: 6.31 N: 14.41

Theoretical value (%); C: 61.86 H: 6.44 N: 14.43

Example 23 1-cyclopropyl-6,8-difluoro-7-[(3-N-methylamino-4-N-methylaminomethylpyrrolidin) -1-yl] -4-oxo-1, Preparation of 4-dihydroquinoline-3-carboxylic acid

295 mg of 1-cyclopropyl-6,7,8-trifluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 3-N-methylamino-4-N-methylaminomethylpyrrolidine 160 mg of trichloride was treated in the same manner as in Example 1 to obtain 397 mg of the target compound.

Elemental Analysis (C ₂₀ H ₂₄ N ₄ O ₃ F ₂ )

Found (%); C: 59.28 H: 5.91 N: 13.71

Theoretical value (%); C: 59.11 H: 5.91 N: 13.79

Example 24 1-cyclopropyl-6,8-difluoro-7-{[3-N- (3'-pyridylmethyl) amino-4-aminomethylpyrrolidin] -1-yl} -4- Preparation of oxo-1,4-dihydroquinoline-3-carboxylic acid

295 mg of 1-cyclopropyl-6,7,8-trifluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 3-N- (p-pyridylmethylamino-4-aminopymethyl 226 mg of lollidine trihydrochloride were treated in the same manner as in Example 1 to obtain 407 mg of the target compound.

Elemental Analysis (C ₂₄ H ₂₃ N ₄ O ₃ F ₂ )

Found (%); C: 63.48 H: 5.11 N: 12.41

Theoretical value (%); C: 63.58 H: 5.08 N: 12.36

Example 25 1-cyclopropyl-6,8-difluoro-7-{[3-amino-4-N- (2-fluoroethyl) aminomethylpyrrolidin] -1-yl} -4-oxo- Preparation of 1,4-dihydroquinoline-3-carboxylic acid

295 mg of 1-cyclopropyl-6,7,8-trifluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 3-amino-4-N- (2-fluoroethyl) aminomethylpy 180 mg of lollidine trichloride was treated in the same manner as in Example 1 to obtain 387 mg of the target compound.

Elemental Analysis (C ₂₀ H ₂₃ N ₄ O ₃ F ₃ )

Measured value (%); C: 56.48 H: 5.40 N: 13.31

Theoretical value (%); C: 56.60 H: 5.42 N: 13.21

Example 26 1-cyclopropyl-6-fluoro-8-methoxy-7-{[3-amino-4-N- (2-fluoroethyl) aminomethylpyrrolidin] -1-yl} -4- Preparation of oxo-1,4-dihydroquinoline-3-carboxylic acid

295 mg of 1-cyclopropyl-6,7-difluoro-8-methoxy-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 3-amino-4-N- (2-fluoroethyl) 180 mg of aminomethylpyrrolidine trichloride was treated in the same manner as in Example 1 to obtain 391 mg of the target compound.

Elemental Analysis (C ₂₁ H ₂₆ N ₄ O ₃ F ₂ )

Measured value (%); C: 60.11 H: 6.31 N: 13.41

Theoretical value (%); C: 60.00 H: 6.19 N: 13.33

Example 27 1-cyclopropyl-6-fluoro-7-{[3-amino-4-N- (2-fluoroethyl) aminomethylpyrrolidin] -1-yl} -4-oxo-1,4 Preparation of -dihydro-1,8-naphthyridine-3-carboxylic acid

264 mg of 1-cyclopropyl-6,7-difluoro-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid and 3-amino-4-N- (2-fluoroethyl 180 mg of aminomethylpyrrolidin trichloride was treated in the same manner as in Example 1 to obtain 376 mg of the target compound.

Elemental Analysis (C ₁₉ H ₂₃ N ₅ O ₃ F ₂ )

Measured value (%); C: 56.11 H: 5.57 N: 17.31

Theoretical value (%); C: 56.02 H: 5.65 N: 17.20

Example 28 1-cyclopropyl-6,8-difluoro-7-{[3-N- (2-fluoroethyl) amino-4-aminomethylpyrrolidin] -1-yl} -4-oxo- Preparation of 1,4-dihydroquinoline-3-carboxylic acid

295 mg of 1-cyclopropyl-6,7,8-trifluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 3-N- (2-fluoroethyl) amino-4-aminomethylpy 180 mg of lollidine trichloride was treated in the same manner as in Example 1 to obtain 387 mg of the target compound.

Elemental Analysis (C ₂₀ H ₂₃ N ₄ O ₃ F ₃ )

Measured value (%); C: 56.48 H: 5.31 N: 13.41

Theoretical value (%); C: 56.60 H: 5.42 N: 13.21

Example 29 1-cyclopropyl-6-fluoro-8-methoxy-7-{[3-N- (2-fluoroethyl) amino-4-aminomethylpyrrolidin] -1-yl} -4- Preparation of oxo-1,4-dihydroquinoline-3-carboxylic acid

295 mg of 1-cyclopropyl-6,7-difluoro-8-methoxy-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 3-N- (2-fluoroethyl) amino-4- 180 mg of aminomethylpyrrolidin trichloride was treated in the same manner as in Example 1 to obtain 367 mg of the target compound.

Elemental Analysis (C ₂₁ H ₂₆ N ₄ O ₃ F ₂ )

Measured value (%); C: 60.08 H: 6.21 N: 13.41

Theoretical value (%); C: 60.00 H: 6.19 N: 13.33

Example 30 1-cyclopropyl-6-fluoro-7-{[3-N- (2-fluoroethyl) amino-4-aminomethylpyrrolidin] -1-yl} -4-oxo-1,4 Preparation of -dihydro-1,8-naphthyridine-3-carboxylic acid

264 mg of 1-cyclopropyl-6,7-difluoro-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid and 3-N- (2-fluoroethyl) amino-4 180 mg of aminomethylpyrrolidine trichloride was treated in the same manner as in Example 1 to obtain 353 mg of the target compound.

Elemental Analysis (C ₁₉ H ₂₃ N ₅ O ₃ F ₂ )

Measured value (%); C: 56.12 H: 5.61 N: 17.26

Theoretical value (%); C: 56.02 H: 5.65 N: 17.20

Example 31 1-cyclopropyl-6-fluoro-7-{[3-methoxyamino-4-aminomethylpyrrolidin] -1-yl} -4-oxo-1,4-dihydro-1, Preparation of 8-naphthyridine-3-carboxylic acid

264 mg of 1-cyclopropyl-6,7-difluoro-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid and 3-methoxyamino-4-aminomethylpyrrolidine 162 mg of trichloride was treated in the same manner as in Example 1 to obtain 301 mg of the target compound.

Elemental Analysis (C ₁₉ H ₂₃ N ₄ O ₄ F)

Measured value (%); C: 58.34 H: 5.81 N: 14.26

Theoretical value (%); C: 58.40 H: 5.89 N: 14.34

Example 32 1-cyclopropyl-6-fluoro-8-methoxy-7-{[3-amino-4- (N-methoxy) aminomethylpyrrolidin] -1-yl} -4-oxo- Preparation of 1,4-dihydroquinoline-3-carboxylic acid

295 mg of 1-cyclopropyl-6,7-difluoro-8-methoxy-4-oxo-1,4-dihydroquinoline-3-carboxylic acid and 3-amino-4- (N-methoxy) aminomethyl 180 mg of pyrrolidine trihydrochloride was treated in the same manner as in Example 1 to obtain 367 mg of the target compound.

Elemental Analysis (C ₂₀ H ₂₅ N ₄ O ₅ F)

Measured value (%); C: 57.14 H: 6.11 N: 13.31

Theoretical value (%); C: 57.08 H: 5.95 N: 13.32

In vitro antimicrobial activity and selective toxicity of the compounds of the present invention and comparative compounds were measured and compared according to the following test examples.

Test Example 1: In vitro antibacterial activity

In vitro antimicrobial testing of the compounds of the present invention and comparative compounds was performed by a 2-fold dilution method using Mueller Hinton agar medium. The solvent used was water or 1N-NaOH aqueous solution, the strain was Hoechst standard strain and the number of inoculated bacteria was 10 ⁷ CFU (Colony Forming Unit) / ml. As a compound of Example 3 and a comparative compound, after incubating at 37 ^° C. for about 18 hours using cyprofloxacin and spafloxacin, the degree of inhibition was measured.

Table 1 shows the in vitro antimicrobial activity of the compound of Example 3 and the comparative compounds of the present invention.

Strain MIC (µg / mL) Compound of Example 3 Cyprofloxacin Spafloxacin Streptococcus pyogenes 308a 0.013 3.125 0.781 Streptococcus pyogenes 77A 0.013 0.781 0.195 Streptococcus faecium MD 8b 0.098 0.391 0.391 Streptococcus aureus SG 511 (Staphylococcus aureus SG 511) 0.013 0.195 0.049 Streptococcus aureus 285 0.049 0.781 0.049 Streptococcus aureus 503 0.025 0.391 0.049

Test Example 2: Selectivity Index

Selective toxicity of the test compound was determined by the inhibitory action of the test compound against topoisomerase II (IC _{100, Topo II} (μg / ml)) and by the test compound against DNA gyrase of E. Coli. The ratio of inhibitory activity (IC _{100, Gyrase} (µg / ml)) is shown. Table 2 shows the selective toxicity of the compounds of Example 3 of the present invention, and the comparative compounds cyprofloxacin and spafloxacin.

IC _{100, Topo II} (µg / mL) IC _{100, Gyrase} (μg / ml) Toxicity Compounds of Example 3 Cyprofloxacin Spafloxacin 1,000500500 0.20.51.0 5,0001,000500

From the results of Tables 1 and 2, it can be seen that the compound of the present invention is excellent in antimicrobial activity and low in cytotoxicity.

As described above, the quinolone derivative having a 3,4-substituted pyrrolidine derivative at the position 7 of the quinolone nucleus, which is prepared by condensation of the quinolonecarboxylic acid and an amine derivative, has excellent antibacterial activity and low selectivity. It is expected to be useful as an antimicrobial agent.

Claims (7)

Quinolonecarboxylic acid derivative of formula (I) or a pharmaceutically acceptable salt thereof: Formula 1

In the above formula, R ₁ means a C ₁ -C ₄ lower alkyl group, or a C ₁ -C ₄ lower alkyl group, a phenyl group or a pyridyl group substituted with halogen; R ₂ , R ₃ , R ₄ and R ₆ each represent hydrogen, a C ₁ -C ₄ lower alkyl group, or a C ₁ -C ₄ lower alkyl group substituted with phenyl or halogen; R ₅ means hydrogen, amino or methyl group; X means nitrogen, methine group (CH) or substituted methine group (CZ), wherein Z is halogen, C ₁ -C ₄ lower alkyl group or alkoxy group, or C ₁ -C ₄ lower alkyl group substituted by halogen or Alkoxy group) X and R ₁ are connected to each other to form COCH ₂ CH (CH ₃ ), CCH ₂ CH ₂ CH (CH ₃ ) or CSCH ₂ CH (CH ₃ ). The method of claim 1, R ₁ is ethyl, cyclopropyl, 2-fluoroethyl or 2,4-difluorophenyl, R ₂ , R ₃ , R ₄ and R ₆ are each hydrogen, benzyl or methyl and R ₅ is hydrogen or amino, X Is nitrogen, methine, fluormethine, chlormethine or methoxymethine, or a pharmaceutically acceptable salt thereof. The method of claim 1, R ₂ , R ₃ , R ₄ and R ₆ are each hydrogen, benzyl or methyl, R ₅ is hydrogen and X and R ₁ are linked to each other with COCH ₂ CH (CH ₃ ) to form a ring, or a pharmaceutically thereof Acceptable salts. The method of claim 1, The following compounds or pharmaceutically acceptable salts thereof: 1-cyclopropyl-6,8-difluoro-7-[(3-N-benzylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline-3 -Carboxylic acid; 1-cyclopropyl-6-fluoro-8-chloro-7-[(3-N-benzylamino-4-aminomethylpyrrodine) -1-yl] -4-oxo-1,4-dihydroquinoline-3 -Carboxylic acid; 1-cyclopropyl-6-fluoro-8-methoxy-7-[(3-N-benzylamino-4-aminomethylpyrrodine) -1-yl] -4-oxo-1,4-dihydroquinoline- 3-carboxylic acid; 1-cyclopropyl-5-amino-6,8-difluoro-7-[(3-N-benzylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-di Hydroquinoline-3-carboxylic acid; 1-cyclopropyl-6-fluoro-7-[(3-N-benzylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline-3-carboxyl mountain; 9-Fluoro-2,3-dihydro-3- (S) -methyl-10-[(3-N-benzylamino-4-aminomethylpyrrolidin) -1-yl] -7-oxo-7H- Pyrido [1.2.3-dec] -1,4-benzooxazine-6-carboxylic acid; 1-cyclopropyl-6-fluoro-7-[(3-N-benzylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydro-1,8-naphthy Lidine-3-carboxylic acid; 1- (2,4-difluorophenyl) -6-fluoro-7-[(3-N-benzylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-di Hydro-1,8-naphthyridine-3-carboxylic acid; 1-cyclopropyl-6,8-difluoro-7-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline-3-carboxyl mountain; 1-cyclopropyl-6-fluoro-8-chloro-7-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline-3-carbox Acid; 1-cyclopropyl-6-fluoro-8-methoxy-7-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline-3- Carboxylic acid; 1-cyclopropyl-5-amino-6,8-difluoro-7-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline- 3-carboxylic acid; 9-Fluoro-2,3-dihydro-3- (S) -methyl-10-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -7-oxo-7H-pyrido [ 1.2.3-de] -1,4-benzooxazine-6-carboxylic acid; 1-cyclopropyl-6-fluoro-7-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydro-1,8-naphthyridine-3 -Carboxylic acid; 1-cyclopropyl-6-fluoro-7-[(3-amino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline-3-carboxylic acid; 1-cyclopropyl-6,8-difluoro-7-[(3-N-methylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline-3 -Carboxylic acid; 1-cyclopropyl-6-fluoro-8-chloro-7-[(3-N-methylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline- 3-carboxylic acid; 1-cyclopropyl-6-fluoro-8-methoxy-7-[(3-N-methylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline 3-carboxylic acid; 1-cyclopropyl-5-amino-6,8-difluoro-7-[(3-N-methylamino-4-aminomethylpyrrolidin) -1-yl] -4-oxo-1,4-di Hydroquinoline-3-carboxylic acid; 9-Fluoro-2,3-dihydro-3- (S) -methyl-10-[(3-N-methylamino-4-aminomethylpyrrolidin) -1-yl] -7-oxo-7H- Pyrido [1.2.3-dec] -1,4-benzooxazine-6-carboxylic acid; 1-cyclopropyl-6,8-difluoro-7-[(3-amino-4-N-methylaminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline-3 -Carboxylic acid; 1-cyclopropyl-6-fluoro-8-methoxy-7-[(3-amino-4-N-methylaminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydroquinoline 3-carboxylic acid; 1-cyclopropyl-6,8-difluoro-7-[(3-N-methylamino-4-N-methylaminomethylpyrrolidin) -1-yl] -4-oxo-1,4-dihydro Quinoline-3-carboxylic acid; 1-cyclopropyl-6,8-difluoro-7-{[3-N- (3'-pyridylmethyl) amino-4-aminomethylpyrrolidin] -1-yl} -4-oxo-1, 4-dihydroquinoline-3-carboxylic acid; 1-cyclopropyl-6,8-difluoro-7-{[3-amino-4-N- (2-fluoroethyl) aminomethylpyrrolidin] -1-yl} -4-oxo-1,4- Dihydroquinoline-3-carboxylic acid; 1-cyclopropyl-6-fluoro-8-methoxy-7-{[3-amino-4-N- (2-fluoroethyl) aminomethylpyrrolidin] -1-yl} -4-oxo-1, 4-dihydroquinoline-3-carboxylic acid; 1-cyclopropyl-6-fluoro-7-{[3-amino-4-N- (2-fluoroethyl) aminomethylpyrrolidin] -1-yl} -4-oxo-1,4-dihydro- 1,8-naphthyridine-3-carboxylic acid; 1-cyclopropyl-6,8-difluoro-7-{[3-N- (2-fluoroethyl) amino-4-aminomethylpyrrolidin] -1-yl} -4-oxo-1,4- Dihydroquinoline-3-carboxylic acid; 1-cyclopropyl-6-fluoro-8-methoxy-7-{[3-N- (2-fluoroethyl) amino-4-aminomethylpyrrolidin] -1-yl} -4-oxo-1, 4-dihydroquinoline-3-carboxylic acid; 1-cyclopropyl-6-fluoro-7-{[3-N- (2-fluoroethyl) amino-4-aminomethylpyrrolidin] -1-yl} -4-oxo-1,4-dihydro- 1,8-naphthyridine-3-carboxylic acid; 1-cyclopropyl-6-fluoro-7-{[3-methoxyamino-4-aminomethylpyrrolidin] -1-yl} -4-oxo-1,4-dihydro-1,8-naphthyridine 3-carboxylic acid; or 1-cyclopropyl-6-fluoro-8-methoxy-7-{[3-amino-4- (N-methoxy) aminomethylpyrrolidin] -1-yl} -4-oxo-1,4- Dihydroquinoline-3-carboxylic acid. The amine derivative of the following general formula (II) and the quinolone derivative of the following general formula (III) are dissolved or suspended in water or an organic solvent in a molar ratio of 1: 1 to 1.5: 1, and a base is added thereto, and then 20 to A process for preparing a compound of formula (I) according to claim 1 comprising the step of reacting at 120 ° C .: Formula 2

Formula 3

Wherein R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and X are as defined in claim 1. Amine derivatives of the general formula (II) Formula 2

Wherein R ₂ , R ₃ , R ₄ and R ₆ are as defined in claim 1. An antimicrobial composition comprising an antimicrobial effective amount of a compound of formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.