RU2298006C2 - Dehalogenated compounds, antibacterial agent and therapeutic agent based on thereof, method for preparing antibacterial or medicinal agent, using dehalogenated compounds for preparing antibacterial or medicinal agent - Google Patents

Abstract

FIELD: organic chemistry, chemical technology, medicine, pharmacy.

SUBSTANCE: invention relates to a synthetic quinolone agent that is effective as medicinal agents, veterinary preparations, drugs used in fishing industry or as antibacterial preserving agents. Invention describes compound represented by the following general formula (I):

as its separate isomers or their mixture, its salt and their hydrates wherein R¹ represents cyclic alkyl group comprising 3-6 carbon atoms that can comprise a substitute chosen from halogen atom; R² represents hydrogen atom; R³ represents hydrogen atom; R⁴ represents hydrogen atom, amino-group, hydroxyl group; A represents nitrogen atom or part of structure as given in the invention claim; each R⁵ and R⁶ represents independently alkyl group comprising 1-6 carbon atoms or hydrogen atom; n means a whole number 1 or 2. Also, invention describes antibacterial agent and therapeutic agent based on compounds of the formula (I) used in treatment of infectious disease, a method for preparing antibacterial agent, a method for preparing a medicinal agent used in treatment of infectious disease and using compound of the formula (I) for preparing an antibacterial agent and using compound of the formula (I) for preparing a medicinal agent used in treatment of infectious disease. Invention provides novel compounds possessing useful biological properties.

EFFECT: improved preparing method of agents, valuable medicinal properties of compounds and agents.

35 cl, 2 tbl, 15 ex

Description

Field of Invention

This invention relates to a synthetic quinolone antibacterial agent that is effective as drugs, veterinary drugs, drugs used in fishing, or as antibacterial preservatives.

State of the art

Since the discovery of norfloxacin, synthetic quinolone antibacterial agents have been improved in terms of antibacterial activity and pharmacokinetic properties and currently a large number of compounds are used in the clinical field as chemotherapeutic agents effective in most systemic infectious diseases.

In recent years, in clinical practice there has been an increase in the number of generations of bacteria having a low sensitivity to synthetic quinolone antibacterial agents. For example, as with Staphylococcus aureus (MRSA) and Streptococcus pneumococcus (PRSP), which are insensitive to β-lactam antibiotics, and Enterococcus (VRE), which is insensitive to aminoglycoside antibacterial agents, the number of cases when gram-positive bacteria are initially resistant drugs other than synthetic quinolone antibacterial agents also become low-sensitive to synthetic quinolone antibacterial agents. Therefore, in clinical practice there is a need for synthetic quinolone antibacterial agents having higher efficacy.

Regarding the side effects of synthetic quinolone antibacterial agents, in addition to the central nervous system stimulation effect, which until now has been a difficulty, seizures are also known to occur as a result of combined use with non-steroidal anti-inflammatory drugs, phototoxicity, and so on, and thus also exists the need to develop safer synthetic quinolone antibacterial agents.

It is known that the structure of substituents in positions 7 and 1 (or provisions corresponding to these provisions; the same statement is used in this description and below) to a large extent affects the antibacterial activity, pharmacokinetic properties and safety of synthetic quinolone antibacterial agents.

It is known that quinolone derivatives having a pyrrolidinyl group with an aminomethyl group in position 3 as a substituent at position 7 of the starting quinolone skeleton possess strong antibacterial activity against gram-negative and gram-positive bacteria. Derivatives of 7- [3- (1-aminomethyl) pyrrolidin-1-yl] quinolone carboxylic acid can be cited as an example [Journal of Medicinal Chemistry, Volume 29, p. 445 (1986)].

Moreover, well-known examples of quinolone carboxylic acid derivatives having a substituent on the carbon atom of the aminomethyl group of the 3- (1-aminomethyl) pyrrolidin-1-yl group include derivatives of 7- [3- (1-aminoethyl) pyrrolidin-1-yl] quinolone carboxylic acid [Journal of Medicinal Chemistry, Volume 36, p. 871 (1993)]; 7- [3- (1-amino-1-methylethyl) pyrrolidin-1-yl] quinolone carboxylic acid derivatives [Journal of Medicinal Chemistry, Volume 37, p. 733 (1994)]; and derivatives of 7- [3- (1-aminoalkyl) pyrrolidin-1-yl] quinolone carboxylic acid [Chemical and Pharmaceutical Bulletin, Volume 42, p. 1442 (1994)] and so on.

However, although the above quinolone derivatives having, as a substituent, a 3- (aminomethyl) pyrrolidin-1-yl group, a 3- (1-aminoethyl) pyrrolidin-1-yl group or a group having a similar structure are compounds which possess strong antibacterial activity, it was found that as a result of low-selective toxicity [see, for example, Journal of Antimicrobial Chemotherapy, Volume 33, p. 685 (1994)], such compounds act not only on bacteria, but also on cells of the eukaryotic organism and they are difficult to use as a medicine x drugs or drugs in veterinary medicine. Therefore, quinolone compounds having such substituents have not yet been used in existing clinical practice.

At the same time, quinolone carboxylic acid derivatives having a 3- (1-aminocycloalkyl) pyrrolidin-1-yl group as a substituent and which are relevant to the present invention have been described in detail in PCT / JP96 / 00208, which describes compounds with the structure shown by formula A or formula B. That is, in the quinolone compounds of formula A, the substituent (X ¹ ) at position 6 is defined as a halogen atom or a hydrogen atom. However, only the quinolone carboxylic acids in which the substituent at position 6 is a fluorine atom or another halogen atom are specifically described in the above patent application. Therefore, PCT / JP96 / 00208 does not provide a specific description regarding quinolone carboxylic acids in which the substituent at position 6 is hydrogen. In addition, this publication does not provide any specific description as an embodiment of the 3- (1-aminocycloalkyl) pyrrolidinyl-substituted-6-hydrogen-substituted-quinolone carboxylic acids to which the present invention relates.

BUT

[In the above formula, AX ¹ represents a halogen atom or a hydrogen atom, and X ² represents a halogen atom. (The definitions of substituents in the compound shown by formula A are those defined in PCT / JP96 / 00208 and do not apply to the definitions of substituents of the present invention, even if the same notation is used.)]

In the above formula, AR ² is represented by formula B:

AT

[In the above formula, B p is an integer of 1-3, q is an integer of 1-3, R ⁹ represents a hydrogen atom or an alkyl group containing 1-6 carbon atoms, R ¹⁰ represents a hydrogen atom, an alkyl group containing 1- 6 carbon atoms, an alkyl group containing 1-6 carbon atoms, which has a hydroxyl group, or an alkyl group containing 1-6 carbon atoms, which has a halogen atom. (The definitions of substituents in the compound shown by formula B are those defined in PCT / JP96 / 00208 and do not apply to the definitions of substituents of the present invention, even if the same notation is used.)]

In addition to the above, an example of a literature source which indicates a quinolone carboxylic acid derivative containing a 3- (1-aminocycloalkyl) pyrrolidin-1-yl group and which is relevant to the present invention is Chemical and Pharmaceutical Bulletin, Volume 42, p. 1442 (1994) . However, in this source there are no descriptions related to 3- (1-aminocycloalkyl) pyrrolidinyl-substituted-6-hydrogen-substituted-quinolone carboxylic acids, which are the compounds of the present invention.

In addition, for example, in PCT / WO99 / 14214, a 6-hydrogen substituted quinolone carboxylic acid derivative is disclosed in which a nitrogen-containing heterocyclic substituent, for example a 3- (1-aminoethyl) pyrrolidin-1-yl group, is introduced via a carbon-nitrogen bond into 7-position of the quinolone skeleton, and which is relevant to this invention. The above application describes the compounds represented by formulas C and D. However, there is no description regarding the 3- (1-aminocycloalkyl) pyrrolidin-1-yl group which is relevant to the present invention as a substituent at position 7 of the quinolone the skeleton shown by the formula C. In addition, in the specified application there is no description regarding 3- (1-aminocycloalkyl) pyrrolidinyl-substituted-6-hydrogen-substituted-quinolone carboxylic acids, which are relevant to the present invention and contain as as a substituent, the above group.

FROM

[In the above formula, CR ¹ represents a cyclic alkyl group containing 3-6 carbon atoms, an alkyl group containing 1 or 2 carbon atoms, a straight chain alkenyl group containing 2-3 carbon atoms, or a branched chain alkyl or alkenyl group containing 3-4 carbon atoms, wherein the alkyl group or cyclic alkyl group may be unsubstituted, or the alkyl group or cyclic alkyl group may be substituted by 1-3 fluorine atoms or a phenyl group which is unsubstituted if substituted 1-3 fluorine atoms or substituted in the 4-position a hydroxyl group, R ⁶ represents a hydrogen atom, a hydroxyl group, an aminocarbonyl group, a bromine atom, a cyano group, an alkyl group having 1 or 2 carbon atoms, or an alkenyl group or alkynyl group, containing 2-4 carbon atoms, and this alkyl group may be unsubstituted or this alkyl group may be substituted by a methyl group or ethyl group which is unsubstituted or substituted by 1-3 fluorine atoms or one hydroxyl a group or an amino group. (The definitions of substituents in the compound shown in Formula C are those defined in PCT / WO99 / 14214 and do not apply to the definitions of substituents of the present invention, even if the same notation is used.)]

In the above formula, X is represented by formula D:

D

[In the above formula D, R ⁷ represents an amino group that is attached to a carbon next to which there is no nitrogen atom of the pyrrolidine ring, and may be unsubstituted or substituted by one or two alkyl groups containing 1-3 carbon atoms, or an aminoalkyl group which attached to carbon on the pyrrolidine ring, and may be unsubstituted or substituted by an alkyl group containing 1-3 carbon atoms, R ⁹ represents a group selected from the group containing a hydrogen atom, an alkyl group containing 1-4 carbon atoms, and lkenyl group and alkynyl group containing 2-6 carbon atoms, and a condensed and spiroalkyl group containing 3-6 carbon atoms, the alkyl parts of these groups may be unsubstituted or substituted by 1-3 fluorine atoms, and the above substituents R ⁷ and R ⁹ can be combined with a pyrrolidine ring to form a ring structure of the condensed or spiro structure type, where this condensed or spirocyclic part is formed of 2-5 carbon atoms and 0 or 1 nitrogen atom. (The definitions of substituents in the compound shown by formula D are those defined in PCT / WO99 / 14214 and do not refer to the definitions of substituents of the present invention, even if the same notation is used.)]

Other examples of literature in which 6-hydrogen substituted-quinolone carboxylic acid derivatives are disclosed that are relevant to the present invention include Journal of Medicinal Chemistry, Volume 39, p. 4952 (1996). However, even in these sources there is no description regarding 3- (1-aminocycloalkyl) pyrrolidinyl-substituted-6-hydrogen-substituted-quinolone carboxylic acids, which are the compounds of the present invention.

Description of the invention

Thorough studies have been conducted to obtain quinolone compounds that have excellent antibacterial activity, high efficacy, and high safety. As a result, it was found that derivatives of 3- (1-aminocycloalkyl) pyrrolidinyl-substituted-6-dehalogen (hydrogen-substituted) quinolone carboxylic acid represented by formula (I) shown below, their salts and their hydrates have strong antibacterial activity against a wide spectrum of gram-negative and gram-positive bacteria and, in particular, have strong antibacterial activity against drug-resistant bacteria, such as gram-positive cocci, including MRSA, PRSP and VRE.

Moreover, it was found that, in addition to such excellent antibacterial activity, the compounds of this invention possess both excellent safety indicators and pharmacokinetic properties and, thus, can be used in clinical cases in which similar effects cannot be achieved when using the compounds preceding this invention, containing substituents of the same structure at position 7 of the original quinolone skeleton. The present invention has been made based on these findings.

A comparison of the derivatives of 6-hydrogen-substituted-quinolone carboxylic acid represented by formula (I), their salts and their hydrates of the present invention and quinolone compounds in which the hydrogen in position 6 of the compound of this invention is replaced by a fluorine atom, shows that both types of compounds have excellent antibacterial activity against a wide range of gram-negative and gram-positive bacteria, including drug-resistant bacteria. However, it was unexpectedly discovered that the derivatives of 6-hydrogen-substituted-quinolone, which are compounds of the present invention, are compounds which, compared with derivatives of 6-fluoro-substituted-quinolone, are extremely safe compounds whose acute toxicity is reduced and significantly reduced. with respect to micronuclear induction, and also exhibit good pharmacokinetic properties, such as improved excretion in urine and so on.

Thus, it was found that even a quinolone compound that contains a 3- (1-aminocycloalkyl) pyrrolidin-1-yl group containing a 3- (aminomethyl) pyrrolidin-1-yl group as a methyl group substituent and which, as mentioned above, known for its low selectivity toxicity, it will unexpectedly be a compound with excellent selective toxicity and a compound with excellent pharmacokinetic properties, if it is a quinolone compound with a structure th of the present invention.

That is, the present invention relates to compounds represented by the following general formula (I), their salts and their hydrates:

[where R ¹ represents an alkyl group containing 1-6 carbon atoms, an alkenyl group containing 2-6 carbon atoms, a haloalkyl group containing 1-6 carbon atoms, a cyclic alkyl group containing 3-6 carbon atoms, which may contain a substituent , an aryl group which may contain a substituent, a heteroaryl group which may contain a substituent, an alkoxy group containing 1-6 carbon atoms, or an alkylamino group containing 1-6 carbon atoms;

R ² represents an alkylthio group containing 1-6 carbon atoms, or a hydrogen atom,

where R ² and the above R ¹ can be combined to form a ring structure by including part of the original skeleton, and the ring thus formed may contain a sulfur atom as the ring atom, and the ring may be substituted with an alkyl group containing 1-6 carbon atoms, which may have a substituent;

R ³ represents a phenylalkyl group consisting of an alkylene group containing 1-6 carbon atoms and a phenyl group, an alkyl group containing 1-6 carbon atoms, an alkoxymethyl group containing 2-7 carbon atoms, a hydrogen atom, a phenyl group, an acetoxymethyl group , pivaloyloxymethyl group, ethoxycarbonyl group, choline group, dimethylaminoethyl group, 5-indanyl group, phthalidinyl group, 5-alkyl-2-oxo-1,3-dioxol-4-ylmethyl group or 3-acetoxy-2-oxobutyl group;

R ⁴ represents an alkyl group containing 1-6 carbon atoms, an alkenyl group containing 2-6 carbon atoms, an alkynyl group containing 2-6 carbon atoms, an alkoxy group containing 1-6 carbon atoms, a hydrogen atom, an amino group, a hydroxyl group, a thiol group or a halogenmethyl group, and

in the above, the amino group may contain one or more substituents selected from the group consisting of an alkyl group containing 1-6 carbon atoms, an acyl group containing 2-5 carbon atoms, and a formyl group;

A represents a nitrogen atom or a part of the structure represented by formula (II):

(where X ¹ represents an alkyl group containing 1-6 carbon atoms, an alkenyl group containing 2-6 carbon atoms, an alkynyl group containing 2-6 carbon atoms, an alkoxy group containing 1-6 carbon atoms, a hydrogen atom, an amino group, an atom halogen, a cyano group, a halogenmethyl group or a halogenomethoxy group,

in the above, the amino group may contain one or more substituents selected from the group consisting of an alkyl group containing 1-6 carbon atoms, an acyl group containing 2-5 carbon atoms, and a formyl group,

where X ¹ and the above R ¹ can be combined to form a ring structure by including part of the original skeleton, the ring thus obtained may contain, as the ring atom, an oxygen atom, a nitrogen atom or a sulfur atom, and this ring may be substituted by an alkyl a group containing 1-6 carbon atoms, which may contain a Deputy);

each R ⁵ and R ⁶ independently represents an alkyl group containing 1-6 carbon atoms, a hydrogen atom or a substituted carboxyl group derived from an amino acid, dipeptide or tripeptide,

where the alkyl group may contain one or more substituents selected from the group comprising an alkylthio group containing 1-6 carbon atoms, an alkoxy group containing 1-6 carbon atoms, a hydroxyl group and a halogen atom; and

n is an integer 1 or 2].

The present invention also relates to each of the following:

to the compound of the above formula (I), its salts or their hydrates, where the compound of formula (I) is a stereochemically pure compound;

to the compound of formula (I), its salts or their hydrates, where n in the formula (I) is 1;

to the compound of formula (I), its salts or their hydrates, where R ³ in the formula (I) represents a hydrogen atom;

to the compound of formula (I), its salts or their hydrates, where R ² in the formula (I) represents a hydrogen atom;

to the compound of formula (I), its salts or their hydrates, where R ⁴ in the formula (I) represents a hydrogen atom;

to the compound of formula (I), its salts or their hydrates, where A in the formula (I) represents a part of the structure represented by formula (II);

to a compound of formula (I), its salts or hydrates thereof, wherein X ¹ in formula (II) is a methoxy group, methyl group, difluoromethoxy group, fluorine atom or chlorine atom;

to the compound of formula (I), its salts or their hydrates, where X ¹ in the formula (II) represents a methoxy group or a methyl group;

to the compound of formula (I), its salts or their hydrates, where each R ⁵ and R ⁶ in the formula (I) represents a hydrogen atom;

to the compound of formula (I), its salts or their hydrates, where one of R ⁵ or R ⁶ in the formula (I) represents a hydrogen atom and the other represents a methyl group;

to a compound of formula (I), its salts or their hydrates, where one of R ⁵ or R ⁶ in formula (I) is a hydrogen atom and the other represents a substituted carboxyl group derived from an amino acid, dipeptide or tripeptide;

to the compound of formula (I), its salts or their hydrates, where each R ⁵ and R ⁶ in the formula (I) represents a combination of a hydrogen atom and a methyl group;

to the compound of formula (I), its salts or their hydrates, where R ⁶ in the formula (I) represents a substituted carboxyl group derived from an amino acid, dipeptide or tripeptide;

to the compound of formula (I), its salts or their hydrates, where a cyclic alkyl group comprising 3-6 carbon atoms, which may contain a substituent, in R ¹ represents a halogenocyclopropyl group;

to the compound of formula (I), its salts or their hydrates, where the halogenocyclopropyl group is a 1,2-cis-2-halogenocyclopropyl group;

to the compound of formula (I), its salts or their hydrates, where the halogenocyclopropyl group is a stereochemically pure substituent;

to the compound of formula (I), its salts or their hydrates, where the halogenocyclopropyl group is a (1R, 2S) -2-halogenocyclopropyl group;

to the compound of formula (I), its salts or their hydrates, where the halogen atom of the halogenocyclopropyl group is a fluorine atom;

to 7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4- oxo-1,8-naphthyridine-3-carboxylic acid, its salts or their hydrates;

to 7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4- oxoquinoline-3-carboxylic acid, its salts or their hydrates;

to 7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -8-chloro-1,4- dihydro-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates;

to 7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -8-fluoro-1,4- dihydro-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates;

to 7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -8-difluoromethoxy-1,4- dihydro-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates;

to 7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8- methoxy-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates;

to 7- [3- (R) - [1- (methylamino) cyclopropyl] pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro -8-methoxy-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates;

to 7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8- methyl 4-oxoquinoline-3-carboxylic acid, its salts or their hydrates;

to 7- [3- (R) - [1- (methylamino) cyclopropyl] pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro -8-methyl-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates;

to 7- [3- (R) - [1- (ethylamino) cyclopropyl] pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro -8-methyl-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates;

to 5-amino-7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4- dihydro-8-fluoro-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates;

to 5-amino-7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4- dihydro-8-methoxy-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates;

to 5-amino-7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4- dihydro-8-methyl-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates;

to 5-amino-7- [3- (R) - [1- (methylamino) cyclopropyl] pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1 , 4-dihydro-8-methoxy-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates;

to 10- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -2,3-dihydro-3- (S) -methyl-7-oxo-7H-pyrido [1,2,3- de] [1,4] benzoxazine-6-carboxylic acid, its salts or their hydrates;

to 1- (cyclopropyl) -8-methyl-7- [3- (R) - [1- (methylamino) cyclopropyl] pyrrolidin-1-yl] -1,4-dihydro-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates;

drugs that contain, as an active component, a compound of formula (I), salts thereof or hydrates thereof;

to an antibacterial agent, which as an active component contains a compound of formula (I), its salts or their hydrates;

to a therapeutic agent for the treatment of an infectious disease, which as an active component contains a compound of formula (I), its salts or their hydrates;

a method for treating a disease, which comprises administering, as an active component, a compound of formula (I), its salts or their hydrates;

to a method of treating an infectious disease, which comprises administering as an active component a compound of formula (I), its salts or their hydrates;

a method for producing a medicament, which comprises preparing a pharmaceutical composition, the composition of which as the active ingredient is a compound of formula (I), its salts or their hydrates;

to a method for producing an antibacterial agent, which comprises preparing a composition, the composition of which as the active component includes a compound of formula (I), its salts or their hydrates;

a method for the preparation of an agent for the treatment of an infectious disease, which comprises preparing a composition, the composition of which as the active component is a compound of formula (I), its salts or their hydrates;

the use of a compound of formula (I), its salts or their hydrates for the manufacture of a medicament;

the use of a compound of formula (I), its salts or their hydrates to obtain an antibacterial agent;

the use of a compound of formula (I), its salts or their hydrates for the manufacture of an agent for the treatment of an infectious disease; and so on.

(Method for carrying out the invention)

The following will describe various substituents of the compounds of this invention represented by formula (I):

(R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , n and A are as defined above) should now be described. (For position 6 of the quinolone starting skeleton or equivalent position in the structural formulas presented in this description, in order to emphasize that a hydrogen atom is attached, and this hydrogen atom is attached to carbon, which is not usually indicated in the usual writing of structural formulas in organic chemistry , is indicated in some cases (as “-H"). However, the structural formulas of this description are indicated in accordance with the rules of the image of structural formulas that are commonly used in the field of organic chemistry, and a hydrogen atom attached to a carbon atom will not always be indicated and will usually be omitted).

The substituent R ¹ represents an alkyl group containing 1-6 carbon atoms, an alkenyl group containing 2-6 carbon atoms, a haloalkyl group containing 1-6 carbon atoms, a cyclic alkyl group containing 3-6 carbon atoms, which may contain a substituent, an aryl group which may contain a substituent, a heteroaryl group which may contain a substituent, an alkoxy group containing 1-6 carbon atoms, or an alkylamino group containing 1-6 carbon atoms.

Where an alkyl group containing 1-6 carbon atoms may be a straight or branched chain alkyl group, preferably an alkyl group containing 1-4 carbon atoms, more preferably an ethyl group. As an alkenyl group containing 2-6 carbon atoms, a vinyl group or a 1-isopropenyl group is preferred. As a haloalkyl group containing 1-6 carbon atoms, a 2-fluoroethyl group is preferred. As a cyclic alkyl group, a cyclopropyl group is particularly preferred. The cyclic alkyl group may contain a substituent, and a halogen atom is preferred as a substituent. As the cyclic alkyl group, a halogenocyclopropyl group which may contain a substituent is preferred, and a fluorine atom is particularly preferred as a halogen atom in this group. As the halogenocyclopropyl group, a monohalogenocyclopropyl group is preferred and a cis-substituted group is even more preferred.

Examples of the aryl group which may contain a substituent include a phenyl group and so on, which may contain 1-3 substituents selected from the group consisting of an alkyl group containing 1-6 carbon atoms, an alkoxy group containing 1-6 carbon atoms, an atom halogen, such as a fluorine atom, a chlorine atom and a bromine atom, a hydroxyl group, an amino group, a nitro group, and so on. (In the case where the aryl group has several substituents, the substituents may be of the same type or may be of different types). More specifically, the phenyl group, 2-fluorophenyl group, 4-fluorophenyl group, 2,4-difluorophenyl group, 2-fluoro-4-hydroxyphenyl group, 3-amino-4,6-difluorophenyl group and 4,6-difluoro-3 a methylaminophenyl group is preferred. As for the aryl group, it refers to a group derived from an aromatic hydrocarbon compound. In addition to the phenyl group, the aryl group can also be a naphthyl group or a tricyclic aryl group containing more rings.

A heteroaryl group is a group derived from a pentacyclic or hexacyclic aromatic heterocyclic compound containing one or more heteroatoms selected from nitrogen atoms, an oxygen atom, and a sulfur atom. Particularly preferred is a pentacyclic or hexacyclic nitrogen-containing heterocyclic substituent containing 1 or 2 nitrogen atoms. For example, a pyridyl group, a pyrimidyl group and so on are desirable. As substituents of such rings, an alkyl group, a halogen atom, and so on are preferred. Particularly preferred is a 6-amino-3,5-difluoro-2-pyridyl group.

As an alkoxy group having 1 to 6 carbon atoms, an alkoxy group derived from the above alkyl group is preferred, and among them, a methoxy group is preferred. As for the alkylamino group containing 1-6 carbon atoms, the alkyl part may be the above alkyl group. As an alkylamino group, a methylamino group is preferred.

As the substituent R ¹ , a cyclic alkyl group or a halogenocycloalkyl group is preferred. Of these, a cyclopropyl group or a 2-halogenocyclopropyl group is preferred. As a halogen atom in the 2nd group, a fluorine atom is preferred.

The substituent R ² represents an alkylthio group containing 1-6 carbon atoms, or a hydrogen atom, R ¹ and R ² can be combined to form a ring structure consisting of a polymethylene chain by including a portion of the starting skeleton (i.e., so as to contain a nitrogen atom, to which R ¹ is attached, and the carbon atom to which R ² is attached). The ring thus formed may contain a sulfur atom as the ring-forming atom, and this ring may also contain an alkyl group or a haloalkyl group containing 1-6 carbon atoms as a substituent. The formed ring may be tetracyclic-hexacyclic in size, and the ring may also be saturated or unsaturated. As a substituent on the formed ring, a methyl or fluoromethyl group is preferred. Examples of the condensed ring structure obtained by this method include the following structures:

(In the above formula, R ⁷ represents an alkyl group containing 1-6 carbon atoms, such as a methyl group, a haloalkyl group containing 1-6 carbon atoms, such as a fluoromethyl group, or a hydrogen atom, and R ⁸ represents a halogen atom, such as a fluorine atom, or a hydrogen atom).

As the substituent R ^{2 of the} compound of formula (I), a hydrogen atom is preferred.

The substituent R ³ represents a phenylalkyl group (aralkyl group) consisting of an alkylene group containing 1-6 carbon atoms, and a phenyl group, or an alkyl group containing 1-6 carbon atoms, an alkoxymethyl group containing 2-7 carbon atoms, a hydrogen atom , phenyl group, acetoxymethyl group, pivaloyloxymethyl group, ethoxycarbonyl group, choline group, dimethylaminoethyl group, 5-indanyl group, phthalidinyl group, 5-alkyl-2-oxo-1,3-dioxol-4-ylmethyl group or 3-acetoxy- 2-oxobutyl hydrochloric group.

In the case where the compound of the present invention is used for antibacterial purposes, it is preferable to use a carboxylic acid compound, where R ³ represents a hydrogen atom. Meanwhile, the quinolone derivative in which the carboxylic acid group forms an ester is effective as an intermediate in the synthesis or as a prodrug. These aspects will be further described in more detail.

R ⁴ represents an alkyl group containing 1-6 carbon atoms, an alkenyl group containing 2-6 carbon atoms, an alkynyl group containing 2-6 carbon atoms, an alkoxy group containing 1-6 carbon atoms, a hydrogen atom, an amino group, a hydroxyl group, a thiol group or a halogenomethyl group, and among the above, the amino group may contain one or more substituents selected from the group containing an alkyl group containing 1-6 carbon atoms, an acyl group containing 2-5 carbon atoms, and a formyl group. In the case where several substituent groups are present, the substituents may be all of the same type or may be many different types.

As an alkyl group, which may be either a straight or branched chain group containing 1 to 6 carbon atoms, a methyl group, an ethyl group, a normal propyl group or an isopropyl group are preferred. As an alkenyl group, which may be either a straight or branched chain group containing 2-6 carbon atoms, a vinyl group is preferred. As the alkynyl group, which may be either a straight or branched chain group containing 2-6 carbon atoms, an ethynyl group is preferred. As the halogen of the halogenomethyl group, a fluorine atom is particularly preferred, and the amount thereof may be 1-3. As an alkoxy group, which may contain 1-6 carbon atoms, a methoxy group is preferred.

The substituent R ⁴ preferably represents a hydrogen atom, an alkyl group or an amino group, and among them, a hydrogen atom, a methyl group or an unsubstituted amino group (—NH ₂ ) is particularly preferred.

In the case where R ⁴ represents an amino group, a hydroxyl group or a thiol group, it may be protected by a protecting group, which is commonly used in the relevant fields.

Examples of such protecting groups include (substituted) alkoxycarbonyl groups such as a tert-butoxycarbonyl group, a 2,2,2-trichloroethoxycarbonyl group, and so on; (substituted) aralkyloxycarbonyl groups such as a benzyloxycarbonyl group, a paramethoxybenzyloxycarbonyl group, a paranitrobenzyloxycarbonyl group and so on; (substituted) acyl groups such as acetyl group, methoxyacetyl group, trifluoroacetyl group, chloroacetyl group, pivaloyl group, formyl group, benzoyl group and so on; (substituted) alkyl groups or (substituted) aralkyl groups such as tert-butyl group, benzyl group, paranitrobenzyl group, parametoxybenzyl group, triphenylmethyl group and so on; (substituted) ethers such as methoxymethyl group, tert-butoxymethyl group, tetrahydropyranyl group, 2,2,2-trichloroethoxymethyl group and so on; and (alkyl and / or aralkyl) substituted silyl groups such as a trimethylsilyl group, an isopropyldimethylsilyl group, a tert-butyldimethylsilyl group, a tribenzylsilyl group, a tert-butyldiphenylsilyl group and so on. (Where "(substituted)" means that the group may contain a substituent). As an intermediate, a compound containing an amino group, a hydroxyl group or a thiol group protected by such a substituent is particularly preferred.

A represents a nitrogen atom or a partial structure expressed by the formula (II):

In the case where A represents part of the structure of formula (II), X ¹ represents an alkyl group containing 1-6 carbon atoms, an alkenyl group containing 2-6 carbon atoms, an alkynyl group containing 2-6 carbon atoms, an alkoxy group containing 1 -6 carbon atoms, a hydrogen atom, an amino group, a halogen atom, a cyano group, a halogenomethyl group or a halogenomethoxy group, and among the above groups, the amino group may contain one or more substituents selected from the group containing an alkyl group containing 1-6 carbon atoms, Cilnius group containing 2-5 carbon atoms, and a formyl group.

As a halogen atom, a fluorine atom, a chlorine atom and a bromine atom are preferred, and a fluorine atom and a chlorine atom are particularly preferred. As the alkyl group, which may be a straight or branched chain group containing 1 to 6 carbon atoms, a methyl group, an ethyl group, a normal propyl group or an isopropyl group are preferred. As an alkenyl group, which may be a straight or branched chain group containing 2-6 carbon atoms, a vinyl group is preferred. As the alkynyl group, which may be straight or branched chain containing 2-6 carbon atoms, an ethynyl group is preferred. As the halogen of the halogenomethyl group, a fluorine atom is particularly preferred, and the amount thereof may be 1-3. As an alkoxy group, which may contain 1-6 carbon atoms, a methoxy group is preferred. As the halogen of the halogenomethoxy group, a fluorine atom is particularly preferred, and the amount thereof may be 1-3.

Among these substituents, an alkyl group or an alkoxy group is preferred. Particularly preferred is a methyl group, ethyl group, methoxy group or difluoromethoxy group.

In addition, this X ¹ and the above R ¹ can be combined to form a ring structure including a polymethylene ring by including a portion of the original skeleton (so as to contain the carbon atom to which X ¹ is attached and the nitrogen atom to which R is attached ¹ ). The ring thus formed, as the atom forming the ring, may contain an oxygen atom, a nitrogen atom or a sulfur atom, and this ring may also contain, as a substituent, an alkyl group containing 1-6 carbon atoms, which, in turn, may have a substituent .

The size of the ring formed may be a pentacyclic-heptacyclic ring, and the atoms forming the ring are not limited to a carbon atom and may include an oxygen atom, a nitrogen atom or a sulfur atom. In addition, this ring may be saturated or unsaturated. The ring thus formed may contain as an substituent an alkyl group containing 1-6 carbon atoms. This alkyl group can be considered as a group similar to the above alkyl group, and is preferably a methyl group. This alkyl group may be substituted by a halogen atom, an alkoxy group, and so on.

As a partial structure that forms the ring structure constructed using X ¹ and R ¹ , the structure of the following formula:

-O-CH ₂ -CH (-CH ₃ ) -

is preferred (the right end in the above formula is attached to a nitrogen atom), and a quinolone skeleton of the following structure is particularly preferred:

If A represents a partial structure of formula (II), preferred combinations of R ⁴ and X ¹ are those in which R ⁴ represents an alkyl group containing 1-6 carbon atoms, an amino group, a hydrogen atom or a hydroxyl group, and X ¹ represents an alkyl group, containing 1-6 carbon atoms, an alkoxy group containing 1-6 carbon atoms, a halogenmethoxy group or a hydrogen atom.

More preferred combinations are those in which R ⁴ represents an amino group, a hydrogen atom, a hydroxyl group or a methyl group, and X ¹ represents a methyl group, a methoxy group, a difluoromethoxy group or a hydrogen atom.

Particularly preferred combinations are those in which R ⁴ represents a hydrogen atom, a hydroxyl group or a methyl group, and X ¹ represents a methyl group or a methoxy group.

Each of the substituents R ⁵ and R ⁶ independently represents an alkyl group containing 1-6 carbon atoms, a hydrogen atom or a substituted carboxyl group derived from an amino acid, dipeptide or tripeptide.

This alkyl group may contain one or more substituents selected from the group consisting of an alkylthio group containing 1-6 carbon atoms, an alkoxy group containing 1-6 carbon atoms, a hydroxyl group and a halogen atom.

As an alkyl group, which may be either a straight or branched chain group containing 1 to 6 carbon atoms, a methyl group, an ethyl group, a normal propyl group or an isopropyl group are preferred.

In the case where the alkyl group contains a hydroxyl group as a substituent, the alkyl group may be a straight or branched chain group containing 1 to 6 carbon atoms, and more preferably, the hydroxyl group is a substituent on the terminal carbon atom of the alkyl group. As the alkyl group containing a hydroxyl group, those groups containing up to 3 carbon atoms are preferred, and hydroxymethyl group, 2-hydroxyethyl group, 2-hydroxypropyl group, 3-hydroxypropyl group and so on are preferred.

In the case where the alkyl group contains a halogen atom as a substituent, the alkyl group may be a straight or branched chain group containing 1-6 carbon atoms, and the halogen atom is preferably a fluorine atom. The number of fluorine atoms can correspond to a state from mono-substituted to perfluoro-substituted. Examples include a monofluoromethyl group, a difluoromethyl group, a trifluoromethyl group, a 2,2,2-trifluoroethyl group, and so on.

In the case where the alkyl group contains an alkylthio group as a substituent, the alkyl group may be a straight or branched chain group containing 1-6 carbon atoms, and the alkylthio group may also be a straight chain or branched group containing 1-6 carbon atoms . As the alkyl group containing the alkylthio group, an alkylthiomethyl group, an alkylthioethyl group or an alkylthiopropyl group is preferred, and more preferably, the alkylthio group is also a group containing 1 to 3 carbon atoms. More preferred examples include a methylthiomethyl group, an ethylthioethyl group, and a methylthioethyl group.

In the case where the alkyl group contains an alkoxy group as a substituent, the alkyl group may be a straight or branched chain group containing 1-6 carbon atoms, and the alkoxy group may also be a straight or branched chain group containing 1-6 carbon atoms. As the alkyl group containing an alkoxy group, an alkoxymethyl group, an alkoxyethyl group or an alkoxypropyl group is preferred, and more preferably, the alkoxy group also represents a group containing up to 3 carbon atoms. More preferred examples include a methoxymethyl group, an ethoxymethyl group, and a methoxyethyl group.

Preferred combinations of R ⁵ and R ⁶ are those in which one represents a hydrogen atom and the other represents a hydrogen atom, an alkyl group or a substituted carboxyl group derived from an amino acid, dipeptide or tripeptide. Among them, a combination is preferred in which one of R ⁵ or R ⁶ represents a hydrogen atom and the other represents a hydrogen atom or an alkyl group. As the alkyl group, a methyl group or an ethyl group is preferred, and a methyl group is particularly preferred. Thus, a combination in which both R ⁵ and R ⁶ represent hydrogen atoms, or a combination in which one of R ⁵ or R ⁶ represents a hydrogen atom and the other represents a methyl group, is particularly preferred. The compound of such a combination may most advantageously exhibit a preferred physiological activity as an antibacterial agent.

As a prodrug, a quinolone derivative is particularly effective in which one of the substituents R ⁵ or R ⁶ represents a hydrogen atom and the other represents a substituted carboxyl group derived from an amino acid, dipeptide or tripeptide. Specific examples related to this derivative will be described below.

The following is a description regarding the halogenocyclopropyl group of R ¹ .

The halogen atom as a substituent is preferably a fluorine atom or a chlorine atom, and a fluorine atom is particularly preferred.

For the stereochemical environment of this group, it is particularly preferred that the halogen atom and the quinolone carboxylic acid group in the cyclopropane ring have a cis configuration. Also, although the cis configuration substituent in this case may take the form of a 2- (S) -halogen-1- (R) -cyclopropyl group or a 2- (R) -halogen-1- (S) -cyclopropyl group, the first of the two is preferred.

It was found that despite the presence of the so-called enantiomorphic isomers due to the cis-2-halogenocyclopropyl group separately from R ¹ , all such isomers have strong antibacterial activity and high safety.

The compound of this invention has excellent characteristics when the substituent of the structure represented by the following formula E is at position 7 of the quinolone starting skeleton, in particular the 1- [2- (S)-fluoro-1- (R) -cyclopropyl] - skeleton 1,4-dihydro-4-oxoquinoline-3-carboxylic acid containing 2- (S) -halogen-1- (R) -cyclopropyl group.

E

For this substituent, as a result of the presence of an asymmetric carbon atom at position 3 of the pyrrolidine ring, there are two optical isomers that are in an enantiomorphic bond relationship. To be more precise, they are listed below:

F

Meanwhile, the correlation of the structural activities of two types of optically active compounds, which are due to the steric configuration of the substituent at position 7 (or its equivalent position) of the 7- [3- (1-aminomethyl) pyrrolidin-1-yl] quinolone carboxylic acid derivative and the correlation of the structural activities of four types of optically active substances which are due to the steric configuration of the substituent at position 7 of the 7- [3- (1-aminoethyl) pyrrolidin-1-yl] quinolone carboxylic acid derivative are described in Journal of Medicinal Chemistry, Volume 36, p. 1442 (1994). In this literature, it is indicated that among such optical isomers, the isomers of the structures shown in the following formula have the greatest antibacterial activity.

Based on steric configurations at position 3 of these pyrrolidine rings, it was suggested that of the two optical isomers shown in the above Formula F, the isomer shown below is more preferred:

Thus, a more preferred compound of the compounds of this invention has the structure represented by the following formula:

Therefore, 3- (1-aminocycloalkyl) pyrrolidinyl substituted-6-hydrogen-substituted-quinolone carboxylic acids represented by formula (I), their salts and their hydrates (especially compounds having a structure in which the 3-position of the above pyrrolidine ring is R- configuration, their salts and their hydrates) have strong antibacterial activity against a wide range of gram-negative and gram-positive bacteria, and, in particular, a characteristic feature of such compounds of this invention is that they have the strength antibacterial activity against antibiotic-resistant bacteria such as gram-positive cocci, including MRSA, PRSP and VRE. In addition, the characteristic features of the compounds of this invention is that they have excellent safety and good pharmacokinetic properties, which makes it possible to use these compounds in clinical cases in which similar effects could not be achieved using the compounds preceding this invention, even if they contain substituents with the same structure.

Such excellent properties of the compounds of this invention are possessed by compounds in which n in the substituent described above is an integer of 1 or 2, and excellent results were especially achieved in compounds in which n is an integer of 1. Therefore, preferred compounds are those in which the cyclic moiety is a tricyclic ring.

In the case where the compound of formula (I) of the present invention has a structure in which diastereomers are present, when administered to a human or animal, it is preferable to administer a compound containing one diastereomer. Used in this description, the term "containing one diastereomer" means not only the case when it is completely free from another diastereomer, but also the case when it is present in a chemically pure form. Thus, in other words, another diastereomer may be present in such an amount that it does not affect the physical constants and physiological activity of the compound.

Also, as used herein, the term “stereochemically pure” means that when a compound or the like is in various isomeric forms due to the presence of asymmetric carbon atoms, the compound contains only one of them. The term "pure" in this case can also be considered as described above.

However, the quinolone carboxylic acid derivative of the present invention can be used both in its free form and as an acid addition salt or a salt of a carboxyl group thereof. Examples of acid addition salts include inorganic acid salts such as hydrochlorides, sulfates, nitrates, hydrobromides, hydroiodides, phosphates, and so on; and organic acid salts such as methanesulfonates, benzenesulfonates, toluenesulfonates (and other sulfonates), acetates, citrates, maleates, fumarates, lactates (and other carboxylates) and so on.

Examples of salts of the carboxyl group include alkali metal salts such as lithium salts, sodium salts, potassium salts and so on; alkaline earth metal salts such as magnesium salts, calcium salts and so on; ammonium salts, triethylamine salts, N-methylglucamine salts, tris- (hydroxymethyl) aminomethane salts; and so on, and they can be either inorganic salts or organic salts.

Also, the free form, acid addition salts and salts of the carboxy group of the quinolone carboxylic acid derivative may be present in the form of hydrates.

However, in the case where the compound of this invention is used for antibacterial purposes, it is preferable to use a carboxylic acid compound in which the substituent R ³ represents a hydrogen atom; a quinolone derivative in which the carboxylic acid group is an ester is effective as an intermediate of a synthesis or prodrug. For example, alkyl esters, benzyl esters, alkoxyalkyl esters, phenylalkyl esters and phenyl esters are effective as synthesis intermediates.

Also, the ester used as a prodrug is an ester that is easily hydrolyzed in a living body and forms the free form of a carboxylic acid, and examples thereof include oxoalkyl esters such as acetoxymethyl ester, pivaloyloxymethyl ester, ethoxycarbonyl ester, choline ester, dimethylaminoethyl ester, 5-indanyl ester, phthalidinyl ester, 5-alkyl-2-oxo-1,3-dioxol-4-ylmethyl ester and 3-acetoxy-2-oxobutyl esters th ester.

In addition, a quinolone derivative in which one of the substituents R ⁵ and R ⁶ represents a hydrogen atom and the other represents a substituted carboxyl group derived from an amino acid, dipeptide or tripeptide, is effective as a prodrug.

The amino acid, dipeptide or tripeptide used to prepare such a prodrug are compounds in which a peptide bond formed by a carboxy group derived from an amino acid, dipeptide or tripeptide and an amino group which is present at the substituent at position 7 of the quinolone carboxylic acid derivative can be easily hydrolyzed in a living organism and to form the free form of an amine, and examples thereof include compounds formed from glycine, alanine, aspartic acid and other amino acids; glycine-glycine, glycine-alanine, alanine-alanine and other dipeptides; and glycine-glycine-alanine, glycine-alanine-alanine and other tripeptides.

The compound of the present invention represented by formula (I) can be obtained in various ways, and in the preferred case, such a compound can be obtained, for example, by reacting a compound represented by formula (III):

[where X ² represents a substituent that functions as a leaving group, such as a substituted or unsubstituted phenylsulfonyl group, a substituted or unsubstituted alkylsulfonyl group containing 1-3 carbon atoms, a fluorine atom, a chlorine atom or a bromine atom;

R ³¹ represents R ³ defined in formula (I), or a boron-containing group represented by formula (IV):

(where Y ³² and Y ³³ may be the same or different from each other, and each represents a fluorine atom or an alkylcarbonyloxy group containing 2-4 carbon atoms); and

R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ and A are as defined in formula (I)]

with a compound of the following formula (V) or an addition salt thereof:

[where each R ⁵¹ and R ⁶¹ independently represents a hydrogen atom, an alkyl group containing 1-6 carbon atoms, or a protecting group for an amino group, or one of R ⁵¹ or R ⁶¹ represents a hydrogen atom, and the other represents a substituted carboxyl group derived from an amino acid, dipeptide or tripeptide, with an amino group, i.e., unsubstituted or protected by a protecting group for an amino group, and

this alkyl group may contain a substituent selected from the group comprising an alkylthio group containing 1-6 carbon atoms, an alkoxy group containing 1-6 carbon atoms, a hydroxyl group, and a halogen atom and

n is as defined in formula (I)]

(in the case when the additive salt is used, the reaction is carried out in the presence of reagents that cause the formation of the free form of the additive salt).

Examples of acid addition salts include inorganic acid salts such as hydrochlorides, sulfates, nitrates, hydrobromides, hydroiodides, phosphates, and so on; and organic acid salts such as methanesulfonates, benzenesulfonates, toluenesulfonates (and other sulfonates), acetates, citrates, maleates, fumarates, lactates (and other carboxylates); and so on.

The reaction can be carried out with or without a solvent. The solvent used in the reaction may be any solvent that does not adversely affect the reaction, and examples thereof include dimethyl sulfoxide, pyridine, acetonitrile, ethanol, chloroform, dimethylformamide, dimethylacetamide, N-methylpyrrolidone, tetrahydrofuran, water, 3-methoxybutanol or mixtures thereof.

The reaction is preferably carried out in the presence of an acid acceptor, such as an inorganic base or an organic base, for example, an inorganic base compound, such as an alkali metal or alkaline earth metal carbonate or bicarbonate, or an organic base compound, such as triethylamine, pyridine, 1,8-diazabicycloundecene , N-methylpiperidine, N, N-diisopropylethylamine and so on.

The reaction temperature should be in the range from room temperature to 200 ° C, and preferably in the range from 25 to 150 ° C. The reaction time should be in the range of 30 minutes to 48 hours, and the reaction is usually carried out for about 30 minutes to 8 hours.

The amino group protecting group may be any protecting group that is commonly used in the relevant field, and examples of the protecting group include alkoxycarbonyl groups which may have a substituent, such as a tert-butoxycarbonyl group, a 2,2,2-trichloroethoxycarbonyl group, and so on; aralkyloxycarbonyl groups which may contain a substituent, such as a benzyloxycarbonyl group, a para-methoxybenzyloxycarbonyl group, a para-nitrobenzyloxycarbonyl group and so on; acyl groups which may contain a substituent, such as an acetyl group, a methoxyacetyl group, a trifluoroacetyl group, a chloroacetyl group, a pivaloyl group, a formyl group, a benzoyl group and so on; alkyl groups which may contain a substituent and aralkyl groups which may contain a substituent, such as a tert-butyl group, a benzyl group, a paranitrobenzyl group, a para-methoxybenzyl group, a triphenylmethyl group and so on; ethers which may have a substituent, such as a methoxymethyl group, a tert-butoxymethyl group, a tetrahydropyranyl group, a 2,2,2-trichloroethoxymethyl group and so on; and substituted silyl groups such as trimethylsilyl group, isopropyl dimethylsilyl group, tert-butyldimethylsilyl group, tribenzylsilyl group, tert-butyldiphenylsilyl group and so on.

In the case where each R ³ and R ³¹ represents an alkyl group containing 1-6 carbon atoms, an alkoxymethyl group containing 2-7 carbon atoms, or a phenylalkyl group (aralkyl group) consisting of an alkylene group containing 1-6 carbon atoms , and a phenyl group, it can be converted to the corresponding carboxylic acid by treatment under acidic or basic conditions, which are commonly used for the hydrolysis of carboxylic acid esters.

In the case where R ³¹ has the structure of formula (IV), it can be converted to the corresponding carboxylic acid by hydrolysis under acidic or basic conditions after the reaction of compound (V) with compound (III).

In the case where it is necessary to remove the protection, the compound of interest represented by the formula (I) can be obtained by removing the protecting group under appropriate conditions suitable for the protecting group.

The compound of formula (V) can be obtained in various ways, and although the method shown in PCT / JP96 / 00208 can be given as an example, the preparation of the compound of formula (V) is not limited to this method.

The compound of formula (V) can be obtained by removing Q from the compound represented by the following formula (VI):

[In the above formula (VI), R ⁵¹² is the same as R ⁵ as defined in formula (I) or represents an amino protecting group, R ⁶ and n are as defined in (I), Q is an amino protecting group,

where the amino protecting group may be selected from the group consisting of a (substituted) alkoxycarbonyl group, a (substituted) aralkyloxycarbonyl group, a (substituted) acyl group, a (substituted) alkyl group, a (substituted) aralkyl group and a substituted silyl group.]

The compound described above may be present in the form of a salt, hydrate or salt hydrate thereof. Examples of acid addition salts include salts of inorganic acids and salts of organic acids. Specific examples thereof include inorganic acid salts such as hydrochlorides, sulfates, hydrobromides, hydroiodides, phosphates, and so on; and organic acid salts such as methanesulfonates, benzenesulfonates, toluenesulfonates (sulfonic acid salts); acetates, citrates, maleates, fumarates, lactates (salts of carboxylic acids); and so on.

When both R ⁵¹² and Q represent amino protecting groups, they may be the same or different from each other. However, it is more preferable in the preparation of compound (I) that each protecting group is removed under different reaction conditions.

Examples of R ⁵¹² and Q which represent amino protecting groups include the groups listed above. That is, examples thereof include a (substituted) alkoxycarbonyl group, a (substituted) aralkyloxycarbonyl group, a (substituted) acyl group, a (substituted) alkyl group, a (substituted) aralkyl group, and a (substituted) silyl group.

Specific examples thereof include (substituted) alkoxycarbonyl groups such as a tert-butoxycarbonyl group, a 2,2,2-trichloroethoxycarbonyl group, and so on; (substituted) aralkyloxycarbonyl groups such as a benzyloxycarbonyl group, a para-methoxybenzyloxycarbonyl group, a para-nitrobenzyloxycarbonyl group and so on; (substituted) acyl groups such as acetyl group, methoxyacetyl group, trifluoroacetyl group, chloroacetyl group, pivaloyl group, formyl group, benzoyl group and so on; (substituted) alkyl groups or (substituted) aralkyl groups such as tert-butyl group, benzyl group, para-nitrobenzyl group, para-methoxybenzyl group, triphenylmethyl group and so on; (substituted) ethers such as methoxymethyl group, tert-butoxymethyl group, tetrahydropyranyl group, 2,2,2-trichloroethoxymethyl group and so on; and substituted silyl groups such as trimethylsilyl group, isopropyl dimethylsilyl group, tert-butyldimethylsilyl group, tribenzylsilyl group, tert-butyldiphenylsilyl group and so on.

Upon receipt of compound (I) using the above compound containing Q as a protective group, it is necessary to carry out the reaction by removing the protective group Q. In this case, its interaction with compound (III) or (V) can be carried out using the so-called reaction in one vessel, or the reaction can be carried out immediately after isolation of compound (V) by removal of the protective group.

As in the case of the compound of formula (V), the compound of formula (VI) can be obtained in various ways, and although the method described in PCT / JP96 / 00208 can be given as an example, this method of obtaining the compound of formula (VI) is not limited .

Cis-2-fluorocyclopropylamine containing a single isomer, which is desirable for the synthesis of a compound of formula (I) containing a single isomer, can be synthesized, for example, by the method described in JP-A-2-231475 (the term “JP” used in this description -A "refers to Unexamined Published Japanese Patent Application). The synthesis of a compound of formula (I) containing a single isomer using an optically active derivative of cis-2-fluorocyclopropylamine obtained by the method described above as a starting compound can be carried out, for example, in accordance with the method described in JP-A-2- 231475.

Specific examples of the compounds of this invention include the following compounds, such as:

7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxo -1,8-naphthyridine-3-carboxylic acid [shown in the following formula];

7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline -3-carboxylic acid [shown in the following formula];

7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -8-chloro-1,4-dihydro -4-oxoquinoline-3-carboxylic acid [shown in the following formula];

7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -8-fluoro-1,4-dihydro -4-oxoquinoline-3-carboxylic acid [shown in the following formula];

7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -8-difluoromethoxy-1,4-dihydro -4-oxoquinoline-3-carboxylic acid [shown in the following formula];

7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methoxy -4-oxoquinoline-3-carboxylic acid [shown in the following formula];

7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl -4-oxoquinoline-3-carboxylic acid [shown in the following formula];

5-amino-7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro -8-fluoro-4-oxoquinoline-3-carboxylic acid [shown in the following formula];

5-amino-7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro -8-methoxy-4-oxoquinoline-3-carboxylic acid [shown in the following formula];

5-amino-7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro -8-methyl-4-oxoquinoline-3-carboxylic acid [shown in the following formula];

10- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -2,3-dihydro-3- (S) -methyl-7-oxo-7H-pyrido [1,2,3-de ] [1,4] benzoxazine-6-carboxylic acid [shown in the following formula];

7- [3- (R) - (1-aminocyclobutyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -8-difluoromethoxy-1,4-dihydro -4-oxoquinoline-3-carboxylic acid [shown in the following formula];

7- [3- (R) - (1-aminocyclobutyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methoxy -4-oxoquinoline-3-carboxylic acid [shown in the following formula];

7- [3- (R) - (1-aminocyclobutyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl -4-oxoquinoline-3-carboxylic acid [shown in the following formula];

7- [3- (R) - [1- (methylamino) cyclopropyl] pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro- 8-methoxy-4-oxoquinoline-3-carboxylic acid [shown in the following formula];

7- [3- (R) - [1- (methylamino) cyclopropyl] pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro- 8-methyl-4-oxoquinoline-3-carboxylic acid [shown in the following formula];

7- [3- (R) - [1- (ethylamino) cyclopropyl] pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro- 8-methyl-4-oxoquinoline-3-carboxylic acid [shown in the following formula];

5-amino-7- [3- (R) - [1- (methylamino) cyclopropyl] pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1, 4-dihydro-8-methyl-4-oxoquinoline-3-carboxylic acid [shown in the following formula];

10- [3- (R) - [1- (methylamino) cyclopropyl] pyrrolidin-1-yl] -2,3-dihydro-3- (S) -methyl-7-oxo-7H-pyrido [1,2, 3-de] [1,4] benzoxazine-6-carboxylic acid [shown in the following formula];

1- (cyclopropyl) -8-methyl-7- [3- (R) - [1- (methylamino) cyclopropyl] pyrrolidin-1-yl] -1,4-dihydro-4-oxoquinoline-3-carboxylic acid [shown in the following formula];

Since the compound of this invention has a strong antibacterial effect, it can be used as a medicine for use in humans, animals and fish, or as preservatives in agricultural chemicals and food products.

In the case where the compound of the present invention is used as a medicament for use in humans, the dose for an adult patient is in the range of 50 mg to 1 g per day, preferably 100 mg to 500 mg per day.

If a medicinal product is used for an animal, its dose varies depending on the purpose of administration (treatment or prophylaxis), the type and size of the particular animal in need of treatment, the specific type of pathogen that caused the infection, and the degree of infection, the daily dose is usually in the range 1 mg to 200 mg, preferably from 5 mg to 100 mg per 1 kg of body weight of each animal.

The daily dose is administered at a time or it is divided into two to four doses taken during the day. In case of urgent need, the daily dose may exceed the amounts indicated above.

Since the compounds of this invention are active against a wide range of microorganisms that cause various infectious diseases, it can be used to treat, prevent or alleviate diseases caused by such pathogens.

Examples of bacteria and bacteroid microorganisms against which these compounds are effective include the genus Staphylococcus, Streptococcus pyogenes, Hemolytic streptococcus, Enterococcus, Pneumococcus, the genus Peptostreptococcus, Neisseria gonorrhoeae, Escherichia coli, the genera Citrobacter, the genus Shibella, the genus Shibella, the genus Shibella, the genus Shibella genus Proteus, Pseudomonas aeruginosa, Haemophilus influenzae, genus Acinetobacter, genus Campylobacter, Chlamydia trachomatis and the like.

Examples of diseases that cause the above pathogens include folliculitis, boils, carbuncles, erysipelas, phlegmon, inflammation of the lymph nodes (lymphadenitis), panaritium, subcutaneous abscess, hydradenitis, multiple inflammation of the sebaceous glands, infectious atheroma, anal abscess, mastitis, superficial secondary infection of wounds after injuries, burns, surgery and so on, pharyngolaryngitis, acute bronchitis, tonsillitis, chronic bronchitis, bronchiectasis, diffuse panbronchitis, a typical infection of a chronic respiratory disease, pneumonia, pyelonephritis, cystitis, prostatitis, epididymitis, gonococcal urethritis, non-gonococcal urethritis, cholecystitis, cholangitis, bacillary dysentery, enteritis, uterine appendage inflammation, uterine biliary arthritis, bartholinitis, , otitis media, sinusitis, amphodontitis, pericoronitis, inflammation of the jaw, peritonitis, endocarditis, sepsis, meningitis, skin infection and so on.

In addition, examples of acid resistant bacteria against which the compounds of this invention are effective include tubercle bacilli [Mycobacterium (hereinafter abbreviated as "M") tuberculosis, M. bovis, M. africanum], atypical acid resistant bacteria [M. kansasii, M. marinum, M. scrofulaceum, M. avium, M. intracellulare, M. xenopi, M. fortuitum, M. chelonae] and so on.

Infections caused by these acid-resistant bacteria are generally classified according to the bacteria causing the infection into three types: tuberculosis, atypical infection caused by the acid-resistant bacteria, and leprosy. In addition to the lungs, an infection caused by a tuberculous bacillus can affect the chest cavity, trachea / bronchi, lymph nodes, usually disseminated, bones and joints, brain membranes / brain, digestive organs (intestines, liver), skin, mammary glands, eyes, middle ear / throat, urinary tract, male genitalia, female genitalia and so on. Lungs most often experience an atypical infection caused by acid-resistant bacteria (infections caused by non-tuberculosis acid-resistant bacteria), and other examples of atypical infection caused by acid-resistant bacteria include local lymphadenitis, soft tissue infections, joint infections, common disseminating infections, and so on.

The compounds of this invention are also effective against various microorganisms that cause infections in animals. Examples of such microorganisms include Escherichia, Salmonella, Pasturella, Haemophilus, Bordetella, Staphylococcus, Mycoplasma and so on.

Specific examples of diseases in birds include escherichiosis, white bacillary diarrhea, avian paratyphoid fever, avian cholera, infectious rhinitis, staphylococcal infection, mycoplasma infection and so on, specific examples of swine diseases include eschirichiosis, salmonellosis, pasturellitis eczema, hematophytosis, hemophilus infection, hematophrenitis, hemoformitis, , mycoplasma infection, and so on, specific examples of cattle diseases include Escherichia coli, salmonellosis, hemorrhagic septicemia, mycoplasma and infection, bovine contagious pleuropneumonia, mastitis and so on, specific examples of dog diseases include coliemia, salmonella infection, hemorrhagic septicemia, pyometra, cystitis, and so on, and specific examples of cat diseases include exudative pleurisy, cystitis, chronic rhinitis, hemophilus infection, feline Mycoplasma infection and so on.

An antibacterial preparation that contains a compound of the present invention can be obtained by selecting the appropriate pharmaceutical preparation in accordance with the method of administration and use, by any method commonly used to produce various pharmaceutical preparations. As for the dosage forms of antibacterial preparations containing the compound of the present invention as its main agent, tablets, powders, granules, capsules, solutions, syrups, elixirs, oily or aqueous suspensions, and so on can be given as examples of oral pharmaceutical preparations.

In the case of injections, a stabilizing agent, an antiseptic agent, a solubilizing agent, and so on, or a solution that may contain these additional agents can be placed in a container and then dried by freeze-drying or the like to a solid preparation for re-use dissolving when used. Also, a single dose may be in one container or multiple doses may be contained in the same container.

Examples of formulations for external use include solutions, suspensions, emulsions, ointments, gels, creams, lotions, sprays, and so on.

Solid preparations, along with the active compound, may contain pharmaceutically acceptable additives. For example, fillers, diluents, binders, disintegrating agents, dissolution enhancing agents, wetting agents, lubricating agents and so on may be selected and mixed as necessary to formulate a preparation.

Examples of forms of liquid preparations include solutions, suspensions, emulsions, and as additives they may contain suspending agents, emulsifying agents, and so on.

Examples of methods for administering a compound of the present invention to animals include a method for direct oral administration or oral administration by admixing into a food, a method for preparing a solution and subsequent oral administration of the solution directly or by adding the solution to drinking water or food, an injection method, and so on.

A pharmaceutical preparation for administering a compound of this invention to animals may optionally be obtained in the form of powders, fine granules, soluble powders, syrups, solutions or injections using methods commonly used in the art.

Examples of pharmaceutical preparations are shown below.

Production Example 1 (capsule):

The compound of example 1
Corn starch
CMC calcium
Hydroxymethyl cellulose
Magnesium stearate
Total 100.0 mg
23.0 mg
22.5 mg
3.0 mg
1.5 mg
150.0 mg

Production Example 2 (solution):

The compound of example 1
Acetic acid or sodium hydroxide
Ethyl para-hydroxybenzoate
Purified water
Total 1-10 g
0.5-2 g
0.1 g
88.9-98.4 g
100 g

An example of obtaining 3 (powder for mixing in the feed):

The compound of example 1
Corn starch
Light Silicon Anhydride
Total 1-10 g
98.5-89.5 g
0.5 g
100 g

The best way of carrying out the invention

A further description of the invention is based on examples and reference examples, however, the invention is not limited to them.

Reference Example 1

Ethyl 2- (2,4-difluoro-3-methylbenzoyl) -3-dimethylaminoacrylate

2,4-Difluoro-3-methylbenzoate (4.97 g, 28.9 mmol) was dissolved in toluene (50 ml) and after addition of N, N-dimethylformamide (0.1 ml) and thionyl chloride (3.16 ml, 43 , 4 mmol) was stirred for 14 hours in an oil bath at 80 ° C. Then the reaction solution was cooled and then concentrated under reduced pressure. After toluene was added to the residue and concentrated again under reduced pressure, the obtained residue was dissolved in tetrahydrofuran (10 ml). This solution, while cooling on ice, was added dropwise to a solution in which ethyl 3-dimethylaminoacrylate (4.97 g, 34.7 mmol) and triethylamine (5.04 ml, 36.1 mmol) were dissolved in tetrahydrofuran (20 ml) . After complete addition, the reaction solution was refluxed for 10 hours. After completion of the reaction, the reaction solution was filtered, the triethylamine hydrochloride salt was removed (washing with diethyl ether), and the filtrate was concentrated under reduced pressure. The resulting residue was applied to a short silica gel chromatography column, and from the n-hexane: ethyl acetate = 1: 1 eluate, 6.70 g (78%) of the title compound was obtained as a yellow powder.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 0.95 (3H, J = 7.08 Hz), 2.18 (3H, t, J = 1.95 Hz), 2.92-3.24 ( 6H, m), 3.99 (2H, q, J = 7.08 Hz), 6.86 (1H, dt, J = 1.22, 8.55 Hz), 7.43 (1H, br s .), 7.75 (1H, s).

IR (KBr, disk): 3055, 2985, 2933, 2875, 2814, 1942, 1693, 1630, 1593, 1477, 1431, 1379, 1277, 1255, 1221 cm ^-1 .

Melting point: 82-84 ° C.

Elemental analysis of C ₁₅ H ₁₇ F ₂ NO ₃ :

Calculated: C 60.60; H 5.76; N 4.71.

Found: C, 60.31; H 5.73; N, 4.73.

Reference Example 2

Ethyl 7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl-4-oxoquinoline-3-carboxylate

Ethyl 2- (2,4-difluoro-3-methylbenzoyl) -3-dimethylaminoacrylate (1.06 g, 3.57 mmol) was dissolved in tetrahydrofuran (15 ml) and, after addition of the salt of para-toluenesulfonic acid (1R, 2S) -2-fluorocyclopropylamine (970 mg, 3.93 mmol), with stirring, a solution was added dropwise in which triethylamine (552 μl, 3.96 mmol) was dissolved in tetrahydrofuran (5 ml) at -15 ° C. After the reaction solution was stirred for 2 hours at room temperature, potassium carbonate (740 mg, 5.36 mmol) and tetrabutylammonium chloride (49.6 mg, 0.179 mmol) were added and the reaction suspension was refluxed with stirring for 5 days. After cooling the reaction solution, tetrahydrofuran was evaporated under reduced pressure. Then dichloromethane (10 ml) was added to the precipitate, and while cooling on ice and stirring, 2 mol / L hydrochloric acid was gradually added dropwise to adjust the pH to about 3. Then, after stirring for 15 minutes at room temperature, it was extracted with dichloromethane (60 ml × 3). After drying over anhydrous sodium sulfate was filtered, the filtrate was concentrated under reduced pressure, the obtained crude crystals were stirred and purified in suspension by ethyl acetate. Thus, 713 mg (65%) of the title compound was obtained as colorless crystals.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.41 (3H, t, J = 7.08 Hz), 1.56-1.62 (2H, t), 2.66 (3H, d, J = 2.69 Hz), 3.85-3.89 (1H, m), 4.39 (2H, q, J = 7.08 Hz), 4.78-4.79 and 4.94-4, 95 (1H, dm, J = 62.74 Hz), 7.13 (1H, t, J = 8.91 Hz), 8.36 (1H, dd, J = 6.71, 8.91 Hz), 8.56 (1H, d, J = 2.93 Hz).

IR (KBr, disk): 3438, 3097, 2983, 2939, 2902, 1907, 1720, 1630, 1593, 1566, 1460, 1429, 1387, 1367, 1311, 1250 cm ^-1 .

Melting point: 187-188 ° C.

Elemental analysis of C ₁₆ H ₁₅ F ₂ NO ₃ :

Calculated: C 62.54; H 4.92; N, 4.56.

Found: C, 62.41; H 4.87; N, 4.53.

Reference Example 3

7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl-4-oxoquinoline-3-carboxylic acid

Ethyl 7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl-4-oxoquinoline-3-carboxylate (1.40 g, 4, 56 mmol) was dissolved in acetic acid (4 ml), and after the addition of concentrated hydrochloric acid (4 ml), it was refluxed for 3 hours. After cooling, the reaction solution was poured into ice water (50 ml) and the precipitated crystals were filtered off. After washing the filtered crystals with excess water, washing with cold ethanol and diethyl ether was carried out in this order, and after drying under reduced pressure, 1.18 g (93%) of the title compound was obtained as a white powder.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.48-1.72 (2H, m), 2.75 (3H, t, J = 2.56 Hz), 4.01 (1H, dd, J = 2.81, 5.25 Hz), 4.83-4.84 and 4.98-5.00 (1H, dm, J = 62.74 Hz), 7.31 (1H, dd, J = 2 20, 8.79 Hz), 8.40-8.44 (1H, m), 8.84 (1H, d, J = 2.69 Hz), 14.50 (1H, br s).

IR (KBr, disk): 3097, 3014, 2956, 2642, 1957, 1728, 1618, 1566, 1508, 1469, 1435, 1389, 1321, 1254, 1200 cm ^-1 .

Melting point: 250-253 ° C.

[α] _D ^24.3 = -50.00 ° (c 0.145, 0.1 mol / L NaOH).

Elemental analysis of C ₁₄ H ₁₁ F ₂ NO ₃ :

Calculated: C 60.22; H 3.97; N, 5.02.

Found: C, 59.92; H 3.98; N, 4.92.

EXAMPLE 1

7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl -4-oxoquinoline-3-carboxylic acid

After adding 3- (R) - [1-tert-butoxycarbonylamino-cyclopropyl] pyrrolidine (185 mg, 817 μmol) and triethylamine (0.50 ml) to dry dimethyl sulfoxide (2 ml), 7-fluoro-1- [2- (S ) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl-4-oxoquinoline-3-carboxylic acid (190 mg, 681 μmol) and refluxed for 17 hours under nitrogen . After concentrating the reaction solution under reduced pressure, the residue was dissolved in chloroform (50 ml). After washing the organic layer with a 10% aqueous citric acid solution (25 ml), the organic layer was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and after adding concentrated hydrochloric acid (5 ml) dropwise to the residue, it was stirred at room temperature while cooling on ice for 30 minutes. Then, 1 mol / L hydrochloric acid (5 ml) was added to the reaction solution, and after washing the yellow acidic aqueous solution with chloroform (20 ml × 3), the pH was adjusted to 12.0 with an aqueous sodium hydroxide solution and the insoluble residue was removed by filtration. After adjusting the pH of the basic aqueous solution to 7.4 with 1 mol / L hydrochloric acid, extraction was carried out with chloroform (100 ml × 4). After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure. The residue thus obtained was purified by preparative chromatography (separation in the lower layer with a mixture of chloroform: methanol: water, 7: 3: 1), recrystallized from ethanol, and dried under reduced pressure. Thus, 112 mg (43%) of the title compound was obtained as yellow crystals.

¹ H NMR (400 MHz, 0.1 mol / L NaOD) δ: 0.54 (4H, d, J = 5.61 Hz, 1.19-1.21 (1H, m), 1.58-1 62 (1H, m), 1.66-1.69 (1H, m), 2.00-2.01 (1H, m), 2.16-2.17 (1H, m), 2.35 (3H, s), 3.16-3.23 (2H, m), 3.37-3.42 (1H, m), 3.54-3.55 (1H, m), 4.04-4 05 (1H, m), 4.94-4.95 and 5.10-5.11 (1H, dm, J = 62.16 Hz), 7.01 (1H, d, J = 8.78 Hz ), 7.95 (1H, d, J = 8.78 Hz), 8.43 (1H, s).

IR (KBr, disk): 3375, 3062, 3006, 2925, 2864, 1728, 1610, 1508, 1475, 1431, 1394, 1348, 1315, 1257 cm ^-1 .

Melting point: 228-230 ° C.

[α] _D ^24.7 = -235.09 ° (c 0.285, 0.1 mol / L NaOH).

Elemental analysis of C ₂₁ H ₂₄ FN ₃ O ₃ :

Calculated: C 65.44; H 6.28; N, 10.90.

Found: C, 65.10; H 6.32; N, 10.76.

Reference Example 4

Ethyl 7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methoxy-4-oxoquinoline-3-carboxylate

A mixture of ethyl (2,4-difluoro-3-methoxy) benzoyl acetate (48.8 g, 189 mmol) synthesized by the method described in PCT / US98 / 19138, triethyl orthoformate (78.6 ml, 472 mmol) and acetic anhydride (250 ml) was stirred under heating for 6 hours in an oil bath with an external temperature of 120 ° C. The reaction solution was then cooled, concentrated under reduced pressure, and cured by drying. Then, the resulting yellow extract was dissolved in toluene (800 ml), para-toluenesulfonic acid salt (1R, 2S) -2-fluorocyclopropylamine (60.1 g, 246 mmol) was added, and a solution was added dropwise with stirring at -15 ° C, which triethylamine (40.8 ml, 293 mmol) is dissolved in toluene (200 ml). After stirring the reaction solution for 4 hours at room temperature, water (500 ml) was added and the organic layer was separated. After washing the organic layer with saturated brine (500 ml × 2), it was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure and dried. The resulting oily substance was then dissolved in 1,4-dioxane (600 ml), and 60% sodium hydride in oil (5.94 g, 242 mmol) was gradually added while cooling with water. After stirring the reaction mixture for 30 minutes at room temperature, it was concentrated under reduced pressure until the volume of the reaction solution became approximately 300 ml. The concentrate thus obtained was slowly poured into 1 mol / L hydrochloric acid, which was stirred and cooled with water, and the precipitated crystals were filtered off. After washing the crystals with a large amount of purified water, a small amount of ethanol and an excess of diethyl ether in this order, the obtained crude crystals were suspended in ethyl acetate and purified. Thus, 49.4 g (80.9%) of the title compound were obtained as colorless crystals.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.42 (3H, t, J = 7.08 Hz), 1.55-1.64 (2H, m), 3.88-3.93 (1H , m), 4.04 (3H, d, J = 1.96 Hz), 4.39 (2H, q, J = 7.08 Hz), 4.78-4.79 and 4.94-4, 95 (1H, dm, J = 62.61 Hz), 7.22 (1H, t, J = 8.79 Hz), 8.24 (1H, dd, J = 5.86, 8.79 Hz), 8.60 (1H, s).

Melting point: 190-193 ° C (decomposition).

Reference Example 5

7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methoxy-4-oxoquinoline-3-carboxylic acid

After dissolving ethyl 7-fluoro-1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methoxy-4-oxoquinoline-3-carboxylate (34.0 g, 105 mmol) in acetic acid (400 ml) and the subsequent addition of concentrated hydrochloric acid (400 ml), the mixture was refluxed for 3 hours. After cooling, the reaction solution was poured into ice water (1500 ml) and the precipitated crystals were filtered off. After washing the filtered crystals with large amounts of water, they were washed with cold ethanol and diethyl ether, in that order, and after drying under reduced pressure, the resulting crude crystals were purified by recrystallization from a mixed solvent of acetonitrile-ethanol and then dried under reduced pressure. Thus, 27.1 g (87.4%) of the title compound was obtained as a white powder.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.45-1.75 (2H, m), 3.87-3.95 (1H, m), 4.03 (3H, d, J = 1, 95 Hz), 4.79-4.81 and 4.97-4.99 (1H, dm, J = 62.68 Hz), 7.30 (1H, t, J = 8.79 Hz), 8, 27 (1H, dd, J = 5.86, 8.79 Hz), 8.76 (1H, s).

Melting point: 261-263 ° C (decomposition).

EXAMPLE 2

7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methoxy -4-oxoquinoline-3-carboxylic acid

After adding 3- (R) - [1-tert-butoxycarbonylamino-cyclopropyl] pyrrolidine (165 mg, 731 μmol) and triethylamine (0.50 ml) to dry dimethyl sulfoxide (2 ml), 7-fluoro-1- [2- (S ) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methoxy-4-oxoquinoline-3-carboxylic acid (180 mg, 609 μmol) and refluxed for 13 hours under nitrogen atmosphere . After concentrating the reaction solution under reduced pressure, the residue was dissolved in chloroform (100 ml). After washing the organic layer with a 10% aqueous citric acid solution (50 ml), the organic layer was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and after adding concentrated hydrochloric acid (5 ml) dropwise to the residue, it was stirred at room temperature for 30 minutes while cooling on ice. Then, 1 mol / L hydrochloric acid (5 ml) was added to the reaction solution, after washing the yellow acidic aqueous solution with chloroform (50 ml × 4), the pH was adjusted to 12.0 with an aqueous solution of sodium hydroxide and insoluble materials were removed by filtration. After adjusting the pH of the basic aqueous solution to 7.4 with 1 mol / L hydrochloric acid, it was extracted with chloroform (100 ml × 4). After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure. The residue thus obtained was purified by preparative chromatography (separation in the lower layer with a mixture of chloroform: methanol: water, 7: 3: 1), recrystallized from isopropyl alcohol, and dried under reduced pressure. Thus, 146 mg (60%) of the title compound was obtained as yellow crystals.

¹ H NMR (400 MHz, 0.1 mol / L NaOD) δ: 0.56 (4H, br s), 1.31-1.37 (1H, m), 1.50-1.56 (1H , m), 1.77-1.78 (1H, m), 2.02-2.04 (1H, m), 2.19-2.21 (1H, m), 3.31-3.32 (1H, m), 3.49-3.51 (3H, m), 3.50 (3H, s), 4.00-4.02 (1H, m), 4.93-4.94 and 5 , 09-5.10 (1H, dm, J = 62.87 Hz), 7.01 (1H, s), 7.90 (1H, d, J = 9.03 Hz), 8.39 (1H, d, J = 3.17 Hz).

IR (KBr, disk): 3373, 3315, 3091, 3003, 2976, 2935, 2856, 1903, 1714, 1618, 1518, 1439, 1371, 1313, 1261, 1219 cm ^-1 .

Melting point: 189-192 ° C.

[α] _D ^24.7 = -50.83 ° (c 0.240, 0.1 mol / L NaOH).

Elemental analysis of C ₂₁ H ₂₄ FN ₃ O ₃ :

Calculated: C 62.83; H 6.03; N, 10.47.

Found: C, 62.50; H 6.04; N, 10.26.

EXAMPLE 3

7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1-cyclopropyl-1,4-dihydro-8-methyl-4-oxoquinoline-3-carboxylic acid

After 3- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] pyrrolidine (132 mg, 585 μmol) and triethylamine (245 μl, 1.76 mmol) were added to dry dimethyl sulfoxide (1 ml), BF ₂ chelate 1- was added cyclopropyl-7-fluoro-1,4-dihydro-8-methyl-4-oxoquinoline-3-carboxylic acid (181 mg, 585 μmol) and stirred at room temperature for 87 hours under nitrogen atmosphere. After adding cold water (50 ml) to the reaction solution, filtering the precipitated solids, the resulting solids were suspended in a mixed solvent (200 ml) ethanol / water (9: 1), triethylamine (1 ml) was added, and then refluxed for 7 o'clock. After concentrating the reaction solution under reduced pressure, the residue was dissolved in chloroform (100 ml), after washing the organic layer with a 10% aqueous citric acid solution (50 ml), the organic layer was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and concentrated hydrochloric acid (2 ml) was added dropwise to the residue under cooling on ice, and stirred at room temperature for 30 minutes. Then, 1 mol / L hydrochloric acid (2 ml) was added to the reaction solution, and after washing the yellow acidic aqueous solution with chloroform (50 ml × 3), the pH was adjusted to 12.0 with an aqueous solution of sodium hydroxide. After adjusting the pH of the basic aqueous solution with 1 mol / L hydrochloric acid to 7.4, it was extracted with chloroform (100 ml × 4). After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure. The residue thus obtained was purified by recrystallization from ethanol and then dried under reduced pressure. Thus, 99.6 mg (46%) of the title compound was obtained as yellow crystals.

¹ H NMR (400 MHz, 0.1 mol / L NaOD) δ: 0.55-0.57 (4H, m), 0.74-0.76 (1H, m), 0.90-0.92 (1H, m), 1.11-1.13 (1H, m), 1.24-1.26 (1H, m), 1.75-1.77 (1H, m), 2.03-2 05 (1H, m), 2.21-2.24 (1H, m), 2.48 (3H, s), 3.29-3.38 (3H, m), 3.53-3.55 (1H, m), 4.10-4.12 (1H, m), 7.07 (1H, s), 7.96 (1H, s), 8.57 (1H, s).

Melting point: 230-233 ° C.

Specific rotation: [α] _D ^24.7 = -169.35 ° (c 0.385, 0.1 mol / L NaOH).

Elemental analysis of C ₂₁ H ₂₅ N ₃ O ₃ :

Calculated: C 68.48%; H 6.86%; N, 11.44%.

Found: C 68.46%; H 6.71%; N, 11.38%.

Reference Example 6

Ethyl 2- (2,6-dichloronicotinoyl) acetate

Monoethyl malonate (6.61 g, 50.0 mmol) was dissolved in anhydrous tetrahydrofuran (100 ml) and, after adding magnesium ethoxide (3.15 g, 28.0 mmol), was stirred at room temperature for 3 hours while cooling on ice. Then, the reaction solution was concentrated under reduced pressure, so that the magnesium salt of monoethyl malonate was obtained. Then, 2,6-dichloronicotinic acid (3.84 g, 20.0 mmol) was dissolved in anhydrous tetrahydrofuran (80 ml) and, after adding 1,1-carbonyldiimidazole (4.87 g, 30.0 mmol), was stirred under ice cooling at room temperature for 1.5 hours. A solution was added dropwise to this solution over 10 minutes while cooling on ice, where the previously prepared magnesium salt of monoethyl malonate was dissolved in anhydrous tetrahydrofuran (160 ml). After complete addition, the temperature was gradually returned to room temperature and then stirred for 4 hours. After adding ethyl acetate (200 ml) to the reaction solution, the organic layer was washed with a 10% aqueous citric acid solution (150 ml × 2), saturated aqueous sodium bicarbonate (150 ml) and saturated brine (150 ml), in that order, and then dried over anhydrous sodium sulfate. After filtration, the residue obtained by concentrating the filtrate under reduced pressure was applied to silica gel for chromatography, and from the n-hexane: ethyl acetate = 3: 1 eluate 4.24 g (81%) of the title compound were obtained as a pale pink oily substance.

¹ H NMR (400 MHz, CHCl ₃ ) δ: 1.12-1.40 (3H, m), 4.08 (1H, s), 4.15-4.35 (2H, m), 5.72 (0.5H, s), 7.37 (1H, dd, J = 14.5, 8.1 Hz), 9.49 (1H, dd, J = 16.4, 8.1 Hz), 12, 52 (0.5H, s).

Reference Example 7

Ethyl 2- (2,6-dichloronicotinoyl) -3- [2- (S)-fluoro-1- (R) -cyclopropylamino] acrylate

Ethyl 2- (2,6-dichloronicotinoyl) acetate (7.03 g, 26.8 mmol) was dissolved in acetic anhydride (30 ml) and, after the addition of triethylorthoformate (60 ml), was stirred in an oil bath at 140 ° C for 2 hours . Then, allowing the reaction solution to cool, it was concentrated under reduced pressure, and after adding toluene (50 ml) to the obtained residue, it was concentrated under reduced pressure. These steps were repeated 3 times, and the resulting residue was dried under reduced pressure. Thus, 8.42 g of ethyl 2- (2,6-dichloronicotinoyl) -3-ethoxyacrylate was obtained as a yellow oily substance.

Further, crude ethyl 2- (2,6-dichloronicotinoyl) -3-ethoxyacrylate (2.11 g, 6.62 mmol) and 2- (S) -fluoro-1- (R) -cyclopropylamine para-toluenesulfonic acid salt ( 2.45 g, 9.91 mmol) was suspended in dichloromethane (30 ml) and triethylamine (2.77 ml, 19.87 mmol) was gradually added dropwise with stirring at -15 ° C. After complete addition, the reaction solution was stirred at room temperature for 15 hours. After adding ethyl acetate (100 ml) to the reaction solution, the organic layer was washed with 10% aqueous citric acid solution (80 ml), saturated aqueous sodium bicarbonate solution (80 ml) and saturated hydrogen chloride solution (80 ml), in that order, and then dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and thus, 2.10 g (90%, 2 processes) of the title compound were obtained as a tan oily substance (E / Z mixture). The resulting material was used in the next reaction without further purification.

¹ H NMR (400 MHz, CHCl ₃ ) δ: 0.85-0.89 (0.7H, m), 1.00-1.04 (2.3H, m), 1.23-1.38 ( 2H, m), 3.01 (1H, m), 3.94-4.05 (2H, m), 4.65-4.84 (1H, m), 7.27-7.31 (1H, m), 7.50-7.57 (1H, m), 8.29-8.38 (1H, m), 11.02 (0.8H, br d, J = 12.5 Hz).

Reference Example 8

Ethyl 7-chloro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylate

After dissolving ethyl 2- (2,6-dichloronicotinoyl) -3- [2- (S) -fluoro-1- (R) -cyclopropylamino] acrylate (2.07 g, 5.97 mmol) in 1,4-dioxane (30 ml), 60% sodium hydride in oil (287 mg, 7.18 mmol) was gradually added to it with stirring at 5 ° C. Then the reaction suspension was stirred for 1.5 hours at room temperature and concentrated under reduced pressure. After the residue was dissolved in chloroform (100 ml), the organic layer was washed with 10% aqueous citric acid solution (80 ml), saturated aqueous sodium bicarbonate (80 ml) and saturated brine (80 ml), in this order, and then dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, diethyl ether was added to the residue thus obtained, and the precipitated crystals were filtered off, washed with diethyl ether, and then dried under reduced pressure at 60 ° C for 16 hours. Thus, 1.25 g (67%) of the title compound was obtained as a white powder.

¹ H NMR (400 MHz, CHCl ₃ ) δ: 1.41 (3H, t, J = 7.1 Hz), 1.59-1.72 (2H, m), 3.58-3.63 (1H , m), 4.41 (2H, q, J = 7.1 Hz), 4.93-5.12 (1H, m), 7.39-7.41 (1H, m), 8.65- 8.68 (2H, m).

MS (m / z): 310 (M ⁺ ).

Reference Example 9

7-chloro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylic acid

A mixture of ethyl 7-chloro-1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylate (567 mg, 1 , 83 mmol), acetic acid (4 ml) and concentrated hydrochloric acid (2 ml) were refluxed for 2.5 hours. After cooling the reaction solution on ice, ice water (20 ml) was poured into the reaction solution, the precipitated crystals were filtered off, and after washing with a large amount of water, a small amount of cold ethanol and an excess of diethyl ether, the crystals were dried under reduced pressure at 80 ° C for 18 hours. Thus, 449 mg (87%) of the title compound was obtained as white needle crystals.

¹ H NMR (400 MHz, CHCl ₃ ) δ: 1.70-1.80 (2H, m), 3.73-3.79 (1H, m), 4.98-5.17 (1H, m) 7.56 (1H, d, J = 8.3 Hz), 8.73 (1H, d, J = 8.5 Hz), 8.97 (1H, s), 14.11 (1H, broad. c)

Melting point: 215-220 ° C.

Specific rotation: [α] _D ^24.5 = + 26.90 ° (c 0.422, 0.1 mol / L NaOH).

Elemental analysis of C ₁₂ H ₈ ClFN ₂ O ₃ :

Calculated: C, 50.99%; H 2.85%; N, 9.91%.

Found: C, 50.90%; H 2.71%; N, 9.91%.

MS (m / z): 282 (M ⁺ ).

EXAMPLE 4

7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxo -1,8-naphthyridine-3-carboxylic acid

After adding 3- (R) - [1- [N- (tert-butoxycarbonyl) amino] cyclopropyl] pyrrolidine (339 mg, 1.50 mmol) and triethylamine (1.39 ml) to dry acetonitrile (10 ml), 7 was added -chloro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylic acid (283 mg, 1.00 mmol) and the mixture was refluxed for 1.5 hours under nitrogen. After concentrating the reaction solution under reduced pressure, the residue was dissolved in a mixed solvent of ethyl acetate (100 ml) and dichloromethane (50 ml), and after washing the organic layer with a 10% aqueous citric acid solution (50 ml) and saturated brine (50 ml), in order, the organic layer was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and after adding concentrated hydrochloric acid (15 ml) to the residue dropwise, while cooling on ice, the solution was stirred at the same temperature for 30 minutes. Then, 1 mol / L hydrochloric acid (10 ml) was added to the reaction solution, and after washing the yellow acidic aqueous solution with chloroform (50 ml × 4), the pH was adjusted to 11.0 with an aqueous solution of sodium hydroxide. After adjusting the pH of the basic aqueous solution with 1 mol / L hydrochloric acid to 7.4, the lower layer (100 ml × 2) was extracted with a mixture of chloroform: methanol: water, 7: 3: 1. Then, the organic layers were combined, and after drying the organic layer over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure. The resulting residue was purified by recrystallization from ethanol and then dried under reduced pressure. Thus, 263 mg (74%) of the title compound was obtained as white crystals.

¹ H NMR (400 MHz, 0.1 mol / L NaOD) δ: 0.49-0.55 (4H, m), 1.50-1.75 (3H, m), 1.95-2.15 (2H, m), 3.00-3.80 (5H, m), 4.90-5.15 (1H, m), 6.38 (1H, dm, J = 9.1 Hz), 8, 01 (1H, d, J = 9.1 Hz), 8.31 (1H, s).

IR (KBr, disk) ν: 3089, 3008, 2871, 1712, 1624, 1566, 1508, 1446, 1379, 1333, 1257, 1187, 1136, 1095, 1024, 985 cm ^-1 .

Melting point: 216-218 ° C.

Specific rotation: [α] _D ^24.5 = + 63.50 ° (c 0.310, 0.1 mol / L NaOH).

Elemental analysis of C ₁₉ H ₂₁ FN ₄ O ₃ :

Calculated: C 61.28%; H 5.68%; N, 15.04%.

Found: C 61.17%; H 5.66%; N, 15.04%.

MS (m / z): 373 ([M + H]) ⁺ .

Reference Example 10

Ethyl 2,4-difluorobenzoyl acetate

Monoethyl malonate (9.25 g, 70.0 mmol) was dissolved in anhydrous tetrahydrofuran in a nitrogen atmosphere (150 ml), and after adding magnesium ethoxide (4.17 g, 36.8 mmol) under ice-cooling, it was stirred at room temperature for 1 hour Then the reaction solution was concentrated under reduced pressure, thereby obtaining the magnesium salt of monoethyl malonate. Then 2,4-difluorobenzoic acid (7.91 g, 50.0 mmol) was dissolved in anhydrous tetrahydrofuran (100 ml) and, after adding 1,1-carbonyldiimidazole (8.52 g, 52.5 mmol), was stirred under ice cooling at room temperature for 1 hour. While cooling on ice, a solution was added dropwise to the solution in which the previously obtained magnesium salt of monoethyl malonate was dissolved in anhydrous tetrahydrofuran (60 ml). After complete addition, the temperature gradually returned to room temperature, and then was stirred for 16 hours. After adding toluene to the reaction solution (100 ml), the organic layer was washed with 10% aqueous citric acid solution (200 ml), saturated aqueous sodium bicarbonate solution (150 ml) and saturated hydrogen chloride solution (150 ml), in that order, and then dried over anhydrous sodium sulfate. After filtering, the precipitate obtained by concentrating under reduced pressure of the filtrate was applied to silica gel for chromatography, and 11.0 g (95%) of the title compound was obtained from the n-hexane: ethyl acetate = 9: 1 eluate as a pale yellow oily substances.

¹ H NMR (400 MHz, CHCl ₃ ) δ: 1.24-1.36 (3H, m), 3.95 (2H × 2/3, d, J = 3.66 Hz), 4.20-4 30 (2H, m), 5.80 (1H × 1/3, s), 6.86-7.02 (2H, m), 7.88-8.04 (1H, m), 12.72 (1H × 1/3, s).

Reference Example 11

Ethyl 7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carboxylate

A mixture of ethyl 2,4-difluorobenzoyl acetate (5.50 g, 24.1 mmol), triethyl orthoformate (8.00 ml, 48.2 mmol) and acetic anhydride (6.8 ml) was stirred in an oil bath at 120 ° C for 16 hours. After the reaction solution was allowed to cool, it was concentrated under reduced pressure, after toluene (30 ml) was added to the resulting residue, it was again concentrated under reduced pressure, and then dried under reduced pressure. Thus, a yellow oily substance was obtained. The substance was dissolved in toluene (100 ml), and after addition of the 2- (S) -fluoro-1- (R) -cyclopropylamine para-toluenesulfonic acid salt (6.46 g, 26.1 mmol), triethylamine was gradually added dropwise (4, 95 ml, 35.6 mmol), stirring at -15 ° C. After complete addition, the reaction solution was stirred at room temperature for 18 hours. After adding water (150 ml) to the reaction solution, it was extracted with ethyl acetate (150 ml × 2). The organic layers were combined and washed with saturated hydrochloric solution (150 ml) and then dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and thereby a brown oily substance was obtained. After the substance was dissolved in dimethylformamide (35 ml), potassium carbonate (6.55 g, 47.4 mmol) was added thereto and then stirred at room temperature for 21 hours. Then, while cooling on ice and stirring, 2 mol / L hydrochloric acid (50 ml) was gradually added to it, and then stirred at room temperature for 6 hours. The precipitated crystals were filtered off and then washed with a large amount of water, a small amount of cold ethanol and an excess of diethyl ether. The resulting crude crystals were then purified by recrystallization from ethyl acetate and then dried under reduced pressure. Thus, 5.92 g (84%) of the title compound was obtained.

¹ H NMR (400 MHz, CHCl ₃ ) δ: 1.41-1.43 (3H, m), 1.69-1.76 (2H, m), 3.39 (1H, br s), 4 , 37-4.43 (2H, m), 5.09 (1H, dm, J = 62.46 Hz), 7.16-7.22 (1H, m), 7.41-7.44 (1H , m), 8.49-8.57 (2H, m).

Melting point: 227-230 ° C.

Reference Example 12

7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carboxylic acid

A mixture of ethyl 7-fluoro-1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carboxylate (4.08 g, 13.9 mmol) , acetic acid (9 ml) and concentrated hydrochloric acid (9 ml) were refluxed for 21 hours. After cooling the reaction solution on ice, ice water (50 ml) was poured into the reaction solution, the precipitated crystals were filtered off, and after washing with a large amount of water, a small amount of cold ethanol and an excess of diethyl ether, the crystals were dried under reduced pressure at 80 ° C for 16 hours. Thus, 3.51 g (95%) of the title compound was obtained as a white powder.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.78-1.84 (2H, m), 3.52-3.53 (1H, m), 5.13 (1H, dm, J = 64, 59 Hz), 7.31-7.36 (1H, m), 7.59 (1H, d, J = 9.26 Hz), 8.54-8.53 (1H, m), 14.55 ( 1H, c).

Melting point: 302-305 ° C.

Specific rotation: [α] _D ^24.3 = + 0.38 ° (c 0.560, 0.1 mol / L NaOH).

EXAMPLE 5

7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline -3-carboxylic acid

After adding 3- (R) - [1- [N- (tert-butoxycarbonyl) amino] cyclopropyl] pyrrolidine (203 mg, 817 μmol) and triethylamine (0.5 ml) to dry dimethyl sulfoxide (1 ml), 7-fluorine was added -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carboxylic acid (197 mg, 743 μmol), and the mixture was refluxed for 15 hours in a nitrogen atmosphere. After the reaction solution was allowed to cool, water (30 ml) was added to the reaction solution while cooling on ice, and the precipitated crystals were filtered off and washed thoroughly with water. After adding concentrated hydrochloric acid (5 ml) to the obtained crystals, it was stirred at the same temperature for 30 minutes while cooling on ice. Then, 1 mol / L hydrochloric acid (10 ml) was added to the reaction solution, and after washing the yellow acidic aqueous solution with chloroform (50 ml × 2), the pH was adjusted to 12.0 with sodium hydroxide solution. After adjusting the pH of the basic aqueous solution to 7.4 with 1 mol / L hydrochloric acid, it was extracted with chloroform (100 ml × 3) and a mixture of chloroform: methanol, 95: 5 (100 ml × 2). The organic layers were then combined and, after drying the organic layer over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure. The resulting residue was purified by recrystallization from ethanol and then dried under reduced pressure. Thus, 203 mg (74%) of the title compound was obtained as yellow crystals.

¹ H NMR (400 MHz, 0.1 mol / L NaOD) δ: 0.50-0.54 (4H, m), 1.63-1.68 (3H, m), 2.00-2.12 (2H, m), 2.94-2.97 (1H, m), 3.16-3.36 (4H, m), 5.16 (1H, dm, J = 62.40 Hz), 6, 43 (1H, s), 6.67 (1H, d, J = 9.02 Hz), 7.97 (1H, d, J = 9.02 Hz), 8.32 (1H, s).

IR (KBr, disk) ν: 3087, 3008, 2951, 2858, 1699, 1681, 1520, 1471, 1458, 1396, 1363, 1371, 1250 cm ^-1 .

Melting point: 251-253 ° C.

Specific rotation: [α] _D ^24.3 = + 41.90 ° (c 0.160, 1 mol / L NaOH).

Elemental analysis of C ₂₀ H ₂₂ FN ₃ O ₃ :

Calculated: C 64.68%; H 5.97%; N, 11.31%.

Found: C 64.69%; H 5.96%; N, 11.25%.

Reference Example 13

BF ₂ chelate 7-bromo-1-cyclopropyl-8-difluoromethoxy-1,4-dihydro-4-oxoquinoline-3-carboxylic acid

Heating and stirring a mixed solution of ethyl 7-bromo-1-cyclopropyl-8-difluoromethoxy-1,4-dihydro-4-oxoquinoline-3-carboxylate (2.01 g, 5.00 mmol), acetic acid (5 ml) and acetic anhydride (5 ml) in an oil bath at 110 ° C, a complex of boron trifluoride-tetrahydrofuran (0.83 ml, 7.50 mmol) was added dropwise over 5 minutes. After stirring the reaction solution at the same temperature for 1.5 hours, excess diethyl ether was added while cooling on ice and the precipitated solids were filtered off (and washed with diethyl ether). After drying under reduced pressure at room temperature, 2.06 g (98%) of the title compound was obtained as a pale gray powder.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 1.15-1.30 (4H, m), 4.43 (1H, m), 7.20 (1H, t, J = 71.9 Hz ), 8.25 (1H, d, J = 8.8 Hz), 8.38 (1H, d, J = 8.8 Hz), 9.36 (1H, s).

EXAMPLE 6

7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1-cyclopropyl-8-difluoromethoxy-1,4-dihydro-4-oxoquinoline-3-carboxylic acid

After adding 3- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] pyrrolidine (338 mg, 1.50 mmol) and triethylamine (209 μl, 1.50 mmol) to dry dimethyl sulfoxide (2 ml), BF ₂ chelate was added 7-bromo-1-cyclopropyl-8-difluoromethoxy-1,4-dihydro-4-oxoquinoline-3-carboxylic acid (422 mg, 1.00 mmol) was stirred at room temperature for 39 hours under nitrogen. After concentrating the reaction solution under reduced pressure, ethanol (20 ml), triethylamine (4 ml) and water (4 ml) were added to the concentrate, and refluxed for 3 hours. After concentrating the reaction solution under reduced pressure, the residue was dissolved in chloroform (100 ml), and after washing the organic layer with a 10% aqueous citric acid solution (50 ml), the organic layer was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and after adding concentrated hydrochloric acid (5 ml) dropwise to the residue, it was stirred at room temperature for 30 minutes while cooling on ice. Then, 3 mol / L hydrochloric acid (30 ml) was added to the reaction solution, and after washing the yellow acidic aqueous solution with chloroform (50 ml × 2), the pH was adjusted to 11.0 with an aqueous solution of sodium hydroxide. After adjusting the pH of the basic aqueous solution to 7.4 with 1 mol / L hydrochloric acid, it was extracted with chloroform (50 ml × 2). After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure. The residue thus obtained was purified by recrystallization from a mixed solvent of ethanol / diethyl ether and then dried under reduced pressure. Thus, 31 mg (8%) of the title compound was obtained as a yellow powder.

¹ H NMR (400 MHz, 0.1 mol / L NaOD) δ: 0.57 (4H, br s), 0.81 (1H, m), 1.03 (1H, m), 1.11 ( 1H, m), 1.25 (1H, m), 1.78 (1H, m), 2.05 (1H, m), 2.22 (1H, m), 3.35-3.60 (4H , m), 4.08 (1H, m), 6.45 (1H, dd, J = 76.3, 73.8 Hz), 7.07 (1H, d, J = 9.3 Hz), 8 00 (1H, d, J = 9.3 Hz), 8.46 (1H, s).

Melting point: 206-207.5 ° C.

Specific rotation: [α] _D ^24.5 = -67.70 ° (c 0.295, 0.1 mol / L NaOH).

Elemental analysis of C ₂₁ H ₂₃ F ₂ N ₃ O ₄ · 0.25CH ₃ CH ₂ OH.

Calculated: C 59.92%; H 5.73%; N, 9.75%.

Found: C 59.85%; H 5.62%; N, 9.68%.

Reference Example 14

Ethyl 6-amino-7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl-5-nitro-4-oxoquinoline-3-carboxylate

Ethyl 6,7-difluoro-1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl-5-nitro-4-oxoquinoline-3-carboxylate (10 04 g, 27.12 mmol) was dissolved in dimethylformamide (150 ml) and a 28% aqueous ammonia solution (32.1 ml) was added dropwise with stirring and cooling on ice. Sealing the reaction solution in a sealed tube, it was stirred at room temperature for 4 hours. The reaction solution was then dissolved in methanol (200 ml) and concentrated under reduced pressure. The resulting residue was purified by recrystallization from a mixed solvent of 2-propanol / chloroform / 28% aqueous ammonia, and after drying under reduced pressure, 7.07 g (71%) of the title compound was obtained as a yellow powder.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.35-1.44 (5H, m), 2.67 (3H, d, J = 3.41 Hz), 3.81-3.87 (1H , m), 4.33-4.41 (3H, m), 4.75-4.78 (0.5H, m), 4.90-4.94 (0.5H, m), 8.47 (1H, d, J = 3.41 Hz).

Elemental analysis of C ₁₆ H ₁₅ F ₂ N ₃ O ₅ :

Calculated: C 52.32%; H 4.12%; N, 11.44%.

Found: C, 52.62%; H 4.16%; N, 11.12%.

Reference Example 15

Ethyl 7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl-5-nitro-4-oxoquinoline-3-carboxylate

Isoamyl nitrite (2.56 ml, 19.1 mmol) was added to dimethylformamide (40 ml) and, while stirring at 65 ° C, a solution was added dropwise over 3 hours in which ethyl 6-amino-7-fluoro-1- [ 2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl-5-nitro-4-oxoquinoline-3-carboxylate (5.00 g, 13.6 mmol) dissolved in dimethylformamide (60 ml). After complete addition, the reaction solution was stirred at 65 ° C for 4 hours, allowed to cool and then poured into water (500 ml). After extraction with chloroform (200 ml × 3), the combined organic layers were washed with 1 mol / L hydrochloric acid (200 ml) and brine (100 ml × 2), in that order, and then dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and the obtained residue was applied to silica gel for chromatography to obtain chloroform: methanol = 30: 1, 2.91 g (61%) of the title compound as a white powder from the eluate.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.37 (3H, t, J = 7.08 Hz), 1.40-1.67 (2H, m), 2.70 (3H, d, J = 2.93 Hz), 3.89-3.93 (1H, m), 4.34-4.40 (2H, m), 4.79-4.83 (0.5H, m), 4, 95-4.98 (0.5H, m), 8.55 (1H, d, J = 2.93 Hz).

Reference Example 16

Ethyl 5-amino-7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl-4-oxoquinoline-3-carboxylate

Ethyl 7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl-5-nitro-4-oxoquinoline-3-carboxylate (2.50 g, 7.10 mmol) was dissolved in acetonitrile (20 ml) and, after the addition of the catalyst, 5% palladium on carbon (water content: 50%, 1.0 g) was stirred at room temperature for 20 hours in a hydrogen atmosphere at atmospheric pressure. After removal of the catalyst by filtration (washing with methanol), the filtrate was concentrated under reduced pressure, the obtained residue was dissolved in ethyl acetate (10 ml) and refluxed for 30 minutes. Then n-hexane (10 ml) was added and, after refluxing for 30 minutes, the reaction solution was left at room temperature. The precipitated crystals were then filtered off, washed with a mixture of n-hexane: ethyl acetate, 1: 1, and dried under reduced pressure at 60 ° C for 16 hours. Thus, 869 mg (38%) of the title compound was obtained as a yellow powder.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.23-1.36 (1H, m), 1.38 (3H, t, J = 7.08 Hz), 1.43-1.56 (1H , m), 2.39 (3H, d, J = 2.20 Hz), 3.70-3.77 (1H, m), 4.37 (2H, q, J = 7.08 Hz), 4 , 71-4.75 (0.5H, m), 4.87-4.90 (0.5H, m), 6.20 (1H, d, J = 11.96 Hz), 8.37 (1H , d, J = 3.43 Hz).

Reference Example 17

5-amino-7-fluoro-1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl-4-oxoquinoline-3-carboxylic acid

Ethyl 5-amino-7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl-4-oxoquinoline-3-carboxylate (735 mg, 2.28 mmol) was dissolved in a mixed solution (8 ml) of acetic acid: water, 1: 1, and after adding concentrated sulfuric acid (90 μl), it was stirred for 4 hours in an oil bath at 120 ° C. After cooling the reaction solution on ice, water (20 ml) was poured into it, and the mixed reaction solution was stirred at room temperature for 2 hours. The precipitated crystals were filtered off, washed with a large amount of water, a small amount of cold ethanol and an excess of diethyl ether, in that order, and then dried under reduced pressure at 80 ° C for 17 hours. Thus, 552 mg (82%) of the title compound was obtained as yellow crystals.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 1.23-1.38 (1H, m), 1.56-1.66 (1H, m), 2.39 (3H, d, J = 2.20 Hz), 4.14-4.22 (1H, m), 4.96-5.00 (0.5H, m), 5.12-5.16 (0.5H, m), 6 50 (1H, d, J = 12.70 Hz); 8.60 (1H, d, J = 3.17 Hz).

Specific rotation: [α] _D ^24.3 = -111.00 ° (c 0.510, 0.1 mol / L NaOH).

EXAMPLE 7

5-amino-7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro -8-methyl-4-oxoquinoline-3-carboxylic acid

After adding 3- (R) - [1- [N- (tert-butoxycarbonyl) amino] cyclopropyl] pyrrolidine (643 mg, 2.55 mmol) and triethylamine (0.5 ml) to the dried dimethyl sulfoxide (1 ml), was added 5-amino-7-fluoro-1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl-4-oxoquinoline-3-carboxylic acid (250 mg, 850 μmol) and stirred at 70 ° C for 37 hours under sealed conditions in a nitrogen atmosphere. Then, the reaction solution was allowed to cool, the reaction solution was concentrated under reduced pressure, the obtained residue was dissolved in ethyl acetate (100 ml) and washed with 10% aqueous citric acid solution (50 ml) and brine (30 ml). The organic layer was then dried over anhydrous sodium sulfate, and after filtration, the filtrate was concentrated under reduced pressure. The resulting residue was then applied to a short column of silica gel for chromatography, and the crude crystals were obtained from chloroform: methanol = 30: 1 eluate. After adding concentrated hydrochloric acid (5 ml) dropwise to the crude crystals while cooling on ice, the solution was stirred at the same temperature for 30 minutes. Then, 1 mol / L hydrochloric acid (10 ml) was added to the reaction solution, and after washing the yellow acidic aqueous solution with chloroform (50 ml × 2), the pH was adjusted to 12.0 with an aqueous solution of sodium hydroxide. After adjusting the pH of the basic aqueous solution to 7.4 with 1 mol / L hydrochloric acid, it was extracted with chloroform (100 ml × 3). The organic layers were then combined, dried over anhydrous sodium sulfate, and then the solvent was evaporated under reduced pressure. The residue thus obtained was purified by recrystallization from ethanol and then dried under reduced pressure. Thus, 32 mg (9%) of the title compound was obtained as yellow crystals.

¹ H NMR (400 MHz, 0.1 mol / L NaOD) δ: 0.54-0.57 (1H, m), 0.60-0.67 (1H, m), 1.23-1.55 (3H, m), 1.74-1.85 (1H, m), 1.97-2.17 (2H, m), 2.33 (3H, s), 3.18-3.27 (2H , m), 3 43-3.47 (1H, m), 3.54-3.63 (1H, m), 3.71-3.78 (1H, m), 4.77-4.79 ( 0.5H, m), 4.93-4.96 (0.5H, m), 6.00 (1H, s), 8.56 (1H, d, J = 3.66 Hz).

IR (KBr, disk) ν: 3402, 3344, 3276, 3097, 2918, 2864, 1724, 1616, 1548, 1506, 1477, 1441, 1408 cm ^-1 .

Melting point: 240-242 ° C (decomposition).

Specific rotation: [α] _D ^23.5 = -225.91 ° (c 0.525, 0.1 mol / L NaOH).

Elemental analysis of C ₂₁ H ₂₅ FN ₄ O ₃ :

Calculated: C 62.99%; H 6.29%; N, 13.99%.

Found: C 62.86%; H 6.38%; N 13.76%.

Reference Example 18

Ethyl 6-amino-7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methoxy-5-nitro-4-oxoquinoline-3-carboxylate

Ethyl 6,7-difluoro-1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methoxy-5-nitro-4-oxoquinoline-3-carboxylate (13 96 g, 36.14 mmol) was dissolved in dimethylformamide (180 ml) and a 28% aqueous ammonia solution (60 ml) was added dropwise with stirring and cooling on ice. After stirring the reaction solution at room temperature for 64 hours, water (100 ml) was added to the reaction solution, and then concentrated under reduced pressure. The resulting water-containing residue was then extracted with ethyl acetate (100 ml × 3) and the organic layers were then combined, washed with water (150 ml × 3) and brine (200 ml), in that order, and then dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, the obtained residue was applied to silica gel for chromatography, and chloroform: methanol = 30: 1 was obtained from the eluate, 8.92 g (64%) of the title compound were obtained as a pale red, oily substance .

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.37 (3H, t, J = 7.0 Hz), 1.49 (1H, ddd, J = 9.5, 6.0, 3.5 Hz ), 1.53-1.58 (1H, m), 3.68 (1H, dt, J = 8.5, 5.5 Hz), 4.11 (3H, d, J = 2.5 Hz) , 4.36 (2H, qq, J = 7.0, 1.5 Hz), 4.51 (2H, broad), 4.83 (1H, dd t, J = 63.3, 5.5, 3.5 Hz), 8.47 (1H, s).

IR (KBr, disk) ν: 3379, 1724, 1608, 1525, 1471, 1323, 1259, 1063 cm ^-1 .

HRMS (FAB): C ₁₆ H ₁₆ F ₂ N ₃ O ₆ (M ⁺ +1).

Calculated: 384,1007.

Found: 384.0974.

Reference Example 19

Ethyl 7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methoxy-5-nitro-4-oxoquinoline-3-carboxylate

Isoamyl nitrile (3.81 g, 32.5 mmol) was added to dimethylformamide (60 ml) and a solution in which ethyl 6-amino-7-fluoro-1- [2- (S) was added dropwise with stirring at 70 ° C. -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methoxy-5-nitro-4-oxoquinoline-3-carboxylate (8.90 g, 23.2 mmol) dissolved in dimethylformamide (120 ml ), within 3 hours. After complete addition, the reaction solution was stirred at 70 ° C for 1 hour, allowed to cool and then poured into water (500 ml). After extraction with ethyl acetate (300 ml × 3), the combined organic layers were washed with 1 mol / L hydrochloric acid (300 ml) and brine (200 ml × 2), in that order, and then dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, the resulting crude crystals were recrystallized with ethanol, and then dried under reduced pressure. Thus, 3.81 g (45%) of the title compound was obtained as yellow crystals.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.37 (3H, t, J = 7.0 Hz), 1.55 (1H, ddd, J = 9.5, 6.0, 3.5 Hz ), 1.59-1.69 (1H, m), 3.93 (1H, dt, J = 8.5, 5.5 Hz), 4.11 (3H, t, J = 2.5 Hz) , 4.37 (2H, dqr, J = 7.0, 1.5 Hz), 4.86 (1H, dddd, J = 63.0, 6.0, 5.5, 3.5 Hz), 7 20 (1H, d, J = 10.0 Hz), 8.55 (1H, d, J = 1.5 Hz).

IR (KBr, disk) ν: 3062, 1722, 1639, 1602, 1544, 1425, 1328, 1259, 1057 cm ^-1 .

Melting point: 167-170 ° C (decomposition).

Elemental analysis: C ₁₆ H ₁₄ F ₂ N ₂ O ₆ :

Calculated: C 52.18%; H 3.89%; N, 7.61%.

Found: C 51.97%; H 3.78%; N 7.56%.

Reference Example 20

Ethyl 5-amino-7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methoxy-4-oxoquinoline-3-carboxylate

Ethyl 7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methoxy-5-nitro-4-oxoquinoline-3-carboxylate (3.71 g, 10.1 mmol) was dissolved in acetonitrile (50 ml) and then, after adding a catalyst, 5% palladium on carbon (water content: 50%, 1.5 g) was stirred at room temperature for 20 hours in a hydrogen atmosphere at atmospheric pressure . After removal of the catalyst by filtration (washing with methanol), the filtrate was concentrated under reduced pressure, the obtained residue was applied to silica gel for chromatography, and 2.68 g (79%) of the title compound was obtained as yellow amorphous from chloroform: methanol = 30: 1 eluate substances.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.38 (3H, t, J = 7.0 Hz), 1.43-1.57 (2H, m), 3.75-3.82 (4H , m), 4.37 (2H, q, J = 7.0 Hz), 4.81 (1H, DDT, J = 62.5, 6.5, 3.5 Hz), 6.24 (1H, d, J = 13.0 Hz), 8.37 (1H, d, J = 2.0 Hz).

HRMS (FAB): C ₁₆ H ₁₇ F ₂ N ₂ O ₄ (M ⁺ +1):

Calculated: 339.1156.

Found: 339.1150.

Reference Example 21

5-amino-7-fluoro-1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methoxy-4-oxoquinoline-3-carboxylic acid

A mixture of ethyl 5-amino-7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methoxy-4-oxoquinoline-3-carboxylate (2, 68 g, 7.92 mmol), acetic acid (20 ml) and concentrated hydrochloric acid (20 ml) were refluxed for 3 hours. After cooling the reaction solution on ice, water (200 ml) was poured into it and the precipitated crystals were filtered off. After washing with a large amount of water, a small amount of cold ethanol and an excess of diethyl ether, in this order, the obtained crude crystals were purified by recrystallization from a mixed solvent of chloroform / methanol and then dried under reduced pressure. Thus, 1.26 g (51%) of the title compound was obtained as yellow crystals.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 1.54-1.64 (2H, m), 3.76 (3H, s), 4.02-4.07 (1H, m), 4 , 89-5.10 (1H, m), 6.59 (1H, d, J = 14.0 Hz), 7.73 (2H, br.), 8.57 (1H, d, J = 1, 5 Hz).

IR (KBr, disk) ν: 3432, 3328, 1699, 1576, 1518, 1281, 1236 cm ^-1 .

Melting point: 291-298 ° C (decomposition).

Specific rotation: [α] _D ^25.0 = + 40.01 ° (c 0.305, 0.1 mol / L NaOH).

Elemental analysis: C ₁₄ H ₁₂ F ₂ N ₂ O ₄ :

Calculated: C 54.20%; H 3.90%; N, 9.03%.

Found: C 54.10%; H 3.86%; N, 9.02%.

EXAMPLE 8

5-amino-7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro -8-methoxy-4-oxoquinoline-3-carboxylic acid

After adding 3- (R) - [1- [N- (tert-butoxycarbonyl) amino] cyclopropyl] pyrrolidine (788 mg, 3.48 mmol) and triethylamine (2 ml) to the dried dimethyl sulfoxide (1 ml), 5-amino -7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methoxy-4-oxoquinoline-3-carboxylic acid (621 mg, 2.00 mol) and stirred at 90 ° C for 168 hours, sealing in a sealed tube in a nitrogen atmosphere. After the reaction solution was allowed to cool and the subsequent concentration of the reaction solution under reduced pressure, the obtained residue was dissolved in chloroform (200 ml) and washed with 10% aqueous citric acid solution (100 ml). Then, the organic layer was dried over anhydrous sodium sulfate, and after filtration, the filtrate was concentrated under reduced pressure. After adding concentrated hydrochloric acid (10 ml) dropwise to the obtained residue while cooling on ice, it was stirred at the same temperature for 30 minutes. Then, 1 mol / L hydrochloric acid (20 ml) was added to the reaction solution, and after washing the yellow acidic aqueous solution with chloroform (50 ml × 3), the pH was adjusted to 12.0 with an aqueous solution of sodium hydroxide. After adjusting the pH of the basic aqueous solution to 7.8 with 1 mol / L hydrochloric acid, it was extracted with chloroform (100 ml × 3). Then, the organic layers were combined, dried over anhydrous sodium sulfate, and then the solvent was evaporated under reduced pressure. The residue thus obtained was purified by recrystallization from a mixed solvent of ethanol and diethyl ether, and then dried under reduced pressure. Thus, 74 mg (9%) of the title compound was obtained as yellow crystals.

¹ H NMR (400 MHz, 0.1 mol / L NaOD) δ: 0.48-0.53 (4H, m), 1.09-1.21 (1H, m), 1.32-1.43 (1H, m), 1.64-1.75 (1H, m), 1.93-2.01 (1H, m), 2.10-2.23 (1H, m), 3.21-3 , 23 (1H, m), 3.21-2.27 (1H, m), 3.36-3.43 (6H, m), 3.79-3.84 (1H, m), 4.85 -4.84 (1H, m), 4.85-5.04 (1H, m), 6.06 (1H, s), 8.01 (1H, d, J = 3.5 Hz).

IR (KBr, disk) ν: 3454, 3410, 1716, 1617, 1577, 1548, 1511, 1232, 1016 cm ^-1 .

Melting point: 172-178 ° C (decomposition).

Elemental analysis: C ₂₁ H ₂₅ FN ₄ O ₄ 0.75 H ₂ O:

Calculated: C 58.66%; H 6.21%; N 13.03%.

Found: C 58.58%; H 6.02%; N, 12.76%.

Reference Example 22

Ethyl 3-dimethylamino-2- (2,3,4-trifluorobenzoyl) acrylate

A mixed solution of 2,3,4-trifluorobenzoic acid (10.3 g, 58.5 mmol), thionyl chloride (6.4 ml, 87.8 mmol) and a catalytic amount of dimethylformamide was refluxed for 30 minutes. Then the reaction solution was allowed to cool, the reaction solution was concentrated under reduced pressure, toluene (30 ml) was added to the residue, and again concentrated under reduced pressure. The resulting residue was then dissolved in tetrahydrofuran (20 ml), and the resulting solution was added to a tetrahydrofuran (40 ml) solution of ethyl β-dimethylaminoacrylate (9.20 g, 64.3 mmol) and triethylamine (10.2 ml, 73.1 mmol) while stirring and cooling on ice. The reaction mixture was then stirred at room temperature for 1.5 hours and then refluxed for 16.5 hours. After the reaction solution was allowed to cool, the precipitated solids were removed by filtration and the filtrate was concentrated under reduced pressure. The residue was then applied to silica gel for chromatography, and 15.1 g (86%) of the title compound was obtained from n-hexane: ethyl acetate = 3: 1 eluate.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.00 (3H, t, J = 7.1 Hz), 2.88 (3H, br s), 3.33 (3H, br s), 4.01 (2H, q, J = 7.1 Hz), 6.95-7.01 (1H, m), 7.34 (1H, br s), 7.80 (1H, s).

Reference Example 23

Ethyl 10-fluoro-2,3-dihydro-3- (S) -methyl-7-oxo-7H-pyrido [1,2,3-de] [1,4] benzoxazine-6-carboxylate

Ethyl 3-dimethylamino-2- (2,3,4-trifluorobenzoyl) acrylate (15.0 g, 49.8 mmol) was dissolved in ethanol (30 ml) and ethanol (10 ml) solution (S) was added dropwise to the solution. -2-amino-1-propanol (4.50 g, 59.8 mmol) with stirring and cooling on ice. After complete addition, the reaction solution was stirred for 1 hour at room temperature and then concentrated under reduced pressure. The resulting residue was dissolved in dimethyl sulfoxide (50 ml) and after adding spray dried calcium fluoride (16 g) was stirred at 120 ° C for 26 hours. After the reaction suspension was allowed to cool, the reaction suspension was concentrated under reduced pressure, chloroform (200 ml) and water (200 ml) were added to the residue, and after separation, the aqueous layer was extracted using chloroform (200 ml). Then, the combined organic layers were washed with brine (100 ml) and then dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, the obtained residue was applied to silica gel for chromatography, and 5.60 g (39%) of the title compound was obtained as a white powder from chloroform: methanol = 50: 1 eluate.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.41 (3H, t, J = 7.1 Hz), 1.61 (3H, d, J = 7.1 Hz), 4.33-4, 44 (5H, m), 7.18 (1H, t, J = 10.0 Hz), 8.06 (1H, dd, J = 10.0, 5.4 Hz), 8.39 (1H, s )

Reference Example 24

10-fluoro-2,3-dihydro-3- (S) -methyl-7-oxo-7H-pyrido [1,2,3-de] [1,4] benzoxazine-6-carboxylic acid

A mixture of ethyl 10-fluoro-2,3-dihydro-3- (S) methyl-7-oxo-7H-pyrido [1,2,3-de] [1,4] benzoxazine-6-carboxylate (5.60 g, 19.2 mmol), acetic acid (25 ml) and concentrated hydrochloric acid (25 ml) were refluxed for 4 hours. After cooling the reaction solution on ice, water (100 ml) was added, the precipitated crystals were filtered off, and then washed with a large amount of water, a small amount of cold ethanol and an excess of diethyl ether, in that order. Then, the crude crystals obtained were suspended in ethanol (40 ml) and stirred at room temperature. The crystals were filtered off, washed with ethanol and then dried under reduced pressure at 80 ° C for 17 hours. Thus, 4.10 g (81%) of the title compound was obtained as a white powder.

¹ H NMR (DMSO-d ₆ ) δ: 1.47 (3H, d, J = 6.8 Hz), 4.44 (1H, d, J = 9.6 Hz), 4.62 (1H, d , J = 9.6 Hz), 4.99 (1H, q-like, J = 6.8 Hz), 7.59 (1H, t, J = 9.1 Hz), 7.95 (1H, dd, J = 9.1, 5.4 Hz, 9.07 (1H, s).

Elemental analysis for C ₁₃ H ₁₀ FNO ₄ :

Calculated: C 59.32%; H 3.83%; N, 7.22%.

Found: C 59.60%; H 3.95%; N, 6.99%.

EXAMPLE 9

10- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -2,3-dihydro-3- (S) -methyl-7-oxo-7H-pyrido [1,2,3-de ] [1,4] benzoxazine-6-carboxylic acid

After adding 3- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] pyrrolidine (252 mg, 1.12 mmol) and triethylamine (0.50 ml) to dried dimethyl sulfoxide (3 ml), 10-fluoro-2 was added , 3-Dihydro-3- (S) -methyl-7-oxo-7H-pyrido [1,2,3-de] [1,4] benzoxazine-6-carboxylic acid (244 mg, 928 μmol) and stirred at heating in an oil bath at 100 ° C for 18 hours in a nitrogen atmosphere. After concentrating the reaction solution under reduced pressure, the residue was dissolved in chloroform (100 ml). After washing the organic layer with a 10% aqueous citric acid solution (50 ml) and brine (50 ml), the organic layer was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and after adding concentrated hydrochloric acid (6 ml), dropwise to the residue, while cooling on ice, it was stirred at room temperature for 30 minutes. After adding 4 ml of water to the reaction solution and washing the acidic aqueous solution with chloroform (10 ml × 3), the pH was adjusted to 12.0 with an aqueous solution of sodium hydroxide. After adjusting the pH of the basic aqueous solution to 7.4 with 1 mol / L hydrochloric acid, it was extracted with chloroform (100 ml × 3). After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure. The residue thus obtained was purified by recrystallization from ethanol and then dried under reduced pressure. Thus, 125 mg (36.5%) of the title compound was obtained as yellow crystals.

¹ H NMR (400 MHz, 0.1 mol / L NaOD) δ: 0.57 (4H, s), 1.54 (3H, d, J = 6.80 Hz), 1.66-1.78 ( 1H, m), 2.01-2.11 (1H, m), 2.19-2.30 (1H, m), 3.38-3.60 (4H, m), 4.25 (1H, d, J = 11.0 Hz), 4.47 (1H, d, J = 11.0 Hz), 4.55-4.63 (1H, m), 7.11 (1H, d, J = 9 , 03 Hz), 7.81 (1H, d, J = 9.03 Hz), 8.32 (1H, s).

IR (KBr, disk) ν: 1634, 1529, 1446, 1429, 1363, 1269, 1227, 798 cm ^-1 .

Melting point: 249-252 ° C (decomposition).

Elemental analysis for C ₂₀ H ₂₃ N ₃ O ₄ · HCl · 0.5 H ₂ O:

Calculated: C 57.90%; H 6.07%; N, 10.13%.

Found: C 57.65%; H 5.87%; N, 9.97%.

EXAMPLE 10

1- [2- (S)-fluoro-1- (R) -cyclopropyl] -8-methyl-7- [3- (R) - [1- (methylamino) cyclopropyl] pyrrolidin-1-yl] -1, 4-dihydro-4-oxoquinoline-3-carboxylic acid

After adding to the dried dimethyl sulfoxide (1 ml) 3- (R) - [1- [N- (tert-butoxycarbonyl) -N- (methyl) amino] cyclopropyl] pyrrolidine (118 mg, 436 μmol) and triethylamine (0.50 ml), 7-fluoro-1- [2- (S) -fluoro-1- (R) -cyclopropyl] -8-methyl-1,4-dihydro-4-oxoquinoline-3-carboxylic acid (122 mg, 436 μmol) and refluxed in an oil bath at 100 ° C for 18 hours under nitrogen. After concentrating the reaction solution under reduced pressure, the residue was dissolved in chloroform (100 ml). After washing the organic layer with a 10% aqueous citric acid solution (100 ml), the organic layer was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and after adding concentrated hydrochloric acid (2 ml) dropwise to the residue, it was stirred at room temperature for 30 minutes while cooling on ice. After adding 1 mol / L hydrochloric acid (2 ml) to the reaction solution and washing the yellow acidic aqueous solution with chloroform (50 ml × 3), the pH was adjusted to 12.0 with an aqueous solution of sodium hydroxide. After adjusting the pH of the basic aqueous solution to 7.4 with 1 mol / L hydrochloric acid, it was extracted with chloroform (100 ml × 3). After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure. The residue thus obtained was then purified by preparative chromatography (separation in the lower layer with a mixture of chloroform: methanol: water, 7: 3: 1), in addition, it was purified by recrystallization from ethanol and then dried under reduced pressure. Thus, 72.8 mg (42%) of the title compound was obtained as yellow crystals.

¹ H NMR (400 MHz, 0.1 mol / L NaOD) δ: 1.21-1.27 (1H, m), 1.50-1.64 (2H, m), 1.99-2.01 (1H, m), 2.34 (3H, s), 2.42 (3H, s), 2.87-2.89 (1H, m), 3.27-3.29 (3H, m), 3.63-3.65 1H, m), 4.06-4.07 (1H, m), 5.05 (1H, d m, J = 63.72 Hz), 7.09 (1H, m) 8.00 (1H, s), 8.44 (1H, s).

IR (KBr, disk) ν: 3348, 3086, 2939, 2844, 2789, 1711, 1614, 1518, 1435, 1354, 1315, 1257, 1221 cm ^-1 .

Melting point: 223-224 ° C.

Specific rotation: [α] _D ^24.7 = -119.66 ° (c 0.295, 0.1 mol / L NaOH).

Elemental analysis for C ₂₂ H ₂₆ FN ₃ O ₃ :

Calculated: C 66.15%; H 6.56%; N, 10.52%.

Found: C 65.92%; H 6.52%; N, 10.40%.

EXAMPLE 11

1- [2- (S) -fluoro-1- (R) -cyclopropyl] -8-methoxy-7- [3- (R) - [1- (methylamino) cyclopropyl] pyrrolidin-1-yl] -1, 4-dihydro-4-oxoquinoline-3-carboxylic acid

After adding to the dried dimethyl sulfoxide (1 ml) 3- (R) - [1- [N- (tert-butoxycarbonyl) -N- (methyl) amino] cyclopropyl] pyrrolidine (102 mg, 379 μmol) and triethylamine (0.50 ml), 7-fluoro-1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methoxy-4-oxoquinoline-3-carboxylic acid (112 mg, 379 μmol) and stirred while heating in an oil bath at 100 ° C for 15 hours in a nitrogen atmosphere. After concentrating the reaction solution under reduced pressure, the residue was dissolved in chloroform (100 ml). After washing the organic layer with a 10% aqueous citric acid solution (100 ml), the organic layer was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and after adding concentrated hydrochloric acid (2 ml) dropwise to the residue, it was stirred at room temperature for 30 minutes while cooling on ice. After adding 1 mol / L hydrochloric acid (2 ml) to the reaction solution and washing the yellow acidic aqueous solution with chloroform (50 ml × 4), the pH was adjusted to 12.0 with an aqueous solution of sodium hydroxide. After adjusting the pH of the basic aqueous solution to 7.4 with 1 mol / L hydrochloric acid, it was extracted with chloroform (100 ml × 3). After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure. The residue thus obtained was then purified by preparative chromatography (separation in the lower layer with a mixture of chloroform: methanol: water, 7: 3: 1), in addition, it was purified by recrystallization from ethanol and then dried under reduced pressure. Thus, 78.3 mg (50%) of the title compound was obtained as yellow crystals.

¹ H NMR (400 MHz, 0.1 mol / L NaOD) δ: 0.57-0.61 (4H, m), 1.33-1.40 (1H, m), 1.56-1.58 (2H, m), 1.99-2.01 (1H, m), 2.34 (3H, s), 2.87-2.89 (1H, m), 3.15-3.17 (1H , m), 3.52-3.54 (3H, m), 3.53 (3H, s), 4.00-4.02 (1H, m), 5.02 (1H, dm, J = 64 , 45 Hz), 7.03 (1H, s), 7.92 (1H, d, J = 7.03 Hz), 8.39 (1H, s).

IR (KBr, disk) ν: 3352, 3095, 3051, 2939, 2837, 2787, 1716, 1699, 1616, 1520, 1439, 1358, 1319, 1259, 1221 cm ^-1 .

Melting point: 213-215 ° C.

Specific rotation: [α] _D ^24.7 = -38.46 ° (c 0.195, 0.1 mol / L NaOH).

Elemental analysis for C ₂₂ H ₂₆ FN ₃ O ₄ :

Calculated: C 63.60%; H 6.31%; N, 10.11%.

Found: C 63.36%; H 6.31%; N, 9.97%.

EXAMPLE 12

5-amino-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -8-methyl-7- [3- (R) - [1- (methylamino) cyclopropyl] pyrrolidin-1-yl ] -1,4-dihydro-4-oxoquinoline-3-carboxylic acid

After adding to the dried dimethyl sulfoxide (4 ml) 3- (R) - [1- [N- (tert-butoxycarbonyl) -N- (methyl) amino] cyclopropyl] pyrrolidine (690 mg, 2.25 mmol) and triethylamine (0 , 50 ml), 5-amino-7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -8-methyl-1,4-dihydro-4-oxoquinoline-3- was added carboxylic acid (250 mg, 850 μmol) and refluxed in an oil bath at 70 ° C for 24 hours in a nitrogen atmosphere. After concentrating the reaction solution under reduced pressure, the residue was dissolved in chloroform (100 ml). After washing the organic layer with a 10% aqueous citric acid solution (100 ml) and brine (100 ml), the organic layer was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and after adding concentrated hydrochloric acid (5 ml) dropwise to the residue, it was stirred at room temperature for 30 minutes while cooling on ice. After adding 1 mol / L hydrochloric acid (2 ml) to the reaction solution and washing the yellow acidic aqueous solution with chloroform (50 ml × 3), the pH was adjusted to 12.0 with an aqueous solution of sodium hydroxide. After adjusting the pH of the basic aqueous solution to 7.4 with 1 mol / L hydrochloric acid, it was extracted with chloroform (100 ml × 3). After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure. The residue thus obtained was then purified by preparative chromatography (separation in the lower layer with a mixture of chloroform: methanol: water, 7: 3: 1), in addition, it was purified by recrystallization from ethanol and then dried under reduced pressure. Thus, 70.0 mg (20%) of the title compound was obtained as yellow crystals.

¹ H NMR (400 MHz, 0.1 mol / L NaOD) δ: 0.56-0.64 (4H, m), 1.21-1.61 (3H, m), 1.92-1.96 (1H, m), 2.22 (3H, s), 2.45 (3H, s), 2.68-2.73 (1H, m), 3.19-3.31 (3H, m), 3.59-3.66 (1H, m), 3.72-3.77 (1H, m), 4.76-4.78 (0.5H, m), 4.98-5.01 (0 , 5H, m), 5.97 (1H, s), 8.55 (1H, d, J = 3.66 Hz).

IR (KBr, disk) ν: 3440, 3329, 3082, 3005, 2964, 2937, 2877, 1716, 1620, 1549, 1506, 1437, 1404 cm ^-1 .

Melting point: 129-131 ° C.

Specific rotation: [α] _D ^22.6 = -291.90 ° (c 0.285, 0.1 mol / L NaOH).

Elemental analysis for C ₂₁ H ₂₅ FN ₄ O ₃ · 0.25 H ₂ O:

Calculated: C 63.07%; H 6.62%; N 13.37%.

Found: C, 62.89%; H 6.42%; N 13.27%.

EXAMPLE 13

2,3-dihydro-3- (S) -methyl-10- [3- (R) - [1- (methylamino) cyclopropyl] pyrrolidin-1-yl] -7-oxo-7H-pyrido [1,2, 3-de] [1,4] benzoxazine-6-carboxylic acid

After adding to the dried dimethyl sulfoxide (1 ml) 3- (R) - [1- [N- (tert-butoxycarbonyl) -N- (methyl) amino] cyclopropyl] pyrrolidine (125 mg, 521 μmol) and triethylamine (0.50 ml) was added 10-fluoro-2,3-dihydro-3- (S) methyl-7-oxo-7H-pyrido [1,2,3-de] [1,4] benzoxazine-6-carboxylic acid (132 mg, 500 μmol) and stirred while heating in an oil bath at 100 ° C for 20 hours in a nitrogen atmosphere. After concentrating the reaction solution under reduced pressure, the residue was dissolved in chloroform (100 ml). After washing the organic layer with a 10% aqueous citric acid solution (50 ml) and brine (50 ml), the organic layer was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and after adding concentrated hydrochloric acid (3 ml) to the residue, it was stirred at room temperature for 30 minutes while cooling on ice. After adding water (3 ml) to the reaction solution and washing the yellow acidic aqueous solution with chloroform (50 ml × 3), the pH was adjusted to 12.0 with an aqueous solution of sodium hydroxide. After adjusting the pH of the basic aqueous solution to 7.4 with 1 mol / L hydrochloric acid, it was extracted with chloroform (100 ml × 3). After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure. The residue thus obtained was then purified by recrystallization from ethanol-aqueous ammonia, and then dried under reduced pressure. Thus, 135 mg (70%) of the title compound was obtained as yellow crystals.

¹ H NMR (400 MHz, 0.1 mol / L NaOD) δ: 0.56-0.60 (4H, m), 1.45-1.50 (1H, m), 1.52 (3H, d , J = 6.59 Hz), 1.99-2.01 (1H, m), 2.32 (3H, s), 2.86-2.88 (1H, m), 3.21-3, 55 (4H, m), 4.22, 4.45 (each 1H, ABq, J = 11.36 Hz), 4.57-4.59 (1H, m), 7.04-7.08 (1H , m), 7.80 (1H, d, J = 9.03 Hz), 8.32 (1H, s).

Melting point: 227-229 ° C.

Specific rotation: [α] _D ^24.7 = -131.00 ° (c 0.200, 0.1 mol / L NaOH).

Elemental analysis for C ₂₁ H ₂₅ N ₃ O ₄ :

Calculated: C 65.78%; H 6.57%; N, 10.96%.

Found: C 65.49%; H 6.55%; N, 10.82%.

EXAMPLE 14

7- [3- (R) - [1- (ethylamino) cyclopropyl] pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -8-methyl-1, 4-dihydro-4-oxoquinoline-3-carboxylic acid

After adding to the dried dimethyl sulfoxide (10 ml) 3- (R) - [1- [N- (tert-butoxycarbonyl) -N- (ethyl) amino] cyclopropyl] pyrrolidine (2.16 g, 8.40 mmol) and triethylamine (4 ml) 7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -8-methyl-1,4-dihydro-4-oxoquinoline-3-carboxylic acid was added (1, 95 g, 7.00 mmol) and was refluxed in an oil bath at 100 ° C for 51 hours in a nitrogen atmosphere. After concentrating the reaction solution under reduced pressure, the residue was dissolved in chloroform (150 ml). After washing the organic layer with a 10% aqueous citric acid solution (100 ml) and brine (100 ml), the organic layer was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and after adding concentrated hydrochloric acid (10 ml) dropwise to the residue, it was stirred at room temperature for 30 minutes while cooling on ice. After adding 1 mol / L hydrochloric acid (20 ml) to the reaction solution and washing the yellow acidic aqueous solution with chloroform (100 ml × 5), the pH was adjusted to 12.0 with an aqueous solution of sodium hydroxide. After adjusting the pH of the basic aqueous solution to 7.4, chloroform (150 ml × 4) was extracted with 1 mol / L hydrochloric acid. After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure.

The residue thus obtained was purified by recrystallization from ethanol and then dried under reduced pressure. Thus, 1.61 g (55%) of the title compound was obtained as yellow crystals.

¹ H NMR (400 MHz, 0.1 mol / L NaOD) δ: 0.57-0.63 (4H, m), 1.04 (3H, t, J = 6.95 Hz), 1.19- 1.25 (1H, m), 1.47-1.64 (2H, m), 1.97-1.98 (1H, m), 2.40 (3H, s), 2.70-2, 73 (2H, m), 2.86-2.87 (1H, m), 3.26-3.28 (3H, m), 3.61-3.63 (1H, m), 4.02- 4.05 (1H, m), 5.03 (1H, dm, J = 64.11 Hz), 7.07 (1H, d, J = 9.26 Hz), 7.98 (2H, d, J = 9.26 Hz), 8.43 (1H, d, J = 3.41 Hz).

IR (KBr, disk) ν: 3294, 2964, 2848, 1699, 1612, 1508, 1473, 1431, 1396, 1389, 1350, 1308, 1261 cm ^-1 .

Melting point: 191-194 ° C.

Specific rotation: [α] _D ^24.3 = -236.55 ° (c 0.145, 0.1 mol / L NaOH).

Elemental analysis for C ₂₃ H ₂₈ FN ₃ O ₃ :

Calculated: C 66.81%; H 6.83%; N, 10.16%.

Found: C, 66.52%; H 6.86%; N, 10.03%.

EXAMPLE 15

1- (Cyclopropyl) -8-methyl-7- [3- (R) - [1- (methylamino) cyclopropyl] pyrrolidin-1-yl] -1,4-dihydro-4-oxoquinoline-3-carboxylic acid

After adding to the dried dimethyl sulfoxide (5 ml) 3- (R) - [1- [N- (tert-butoxycarbonyl) -N- (methyl) amino] cyclopropyl] pyrrolidine (880 mg, 3.25 mmol) and triethylamine (1 , 0 ml) 1-cyclopropyl-7-fluoro-8-methyl-1,4-dihydro-4-oxoquinoline-3-carboxylic acid (425 mg, 1.63 mmol) was added and refluxed in an oil bath at 70 ° C for 38 hours in a nitrogen atmosphere. After concentrating the reaction solution under reduced pressure, the residue was dissolved in ethyl acetate (200 ml). After washing the organic layer with a 10% aqueous citric acid solution (100 ml), the organic layer was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and after adding concentrated hydrochloric acid (6 ml), dropwise to the residue, while cooling on ice, it was stirred at room temperature for 30 minutes. After adding 1 mol / L hydrochloric acid (12 ml) to the reaction solution and washing the yellow acidic aqueous solution with chloroform (50 ml × 3), the pH was adjusted to 12.0 with an aqueous solution of sodium hydroxide. After adjusting the pH of the basic aqueous solution to 7.4 with 1 mol / L hydrochloric acid, it was extracted with chloroform (100 ml × 3). After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure. The residue thus obtained was then purified by recrystallization from a mixed solvent of methanol-1/2-propanol and then dried under reduced pressure. Thus, 331 mg (53%) of the title compound was obtained as yellow crystals.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 0.57-0.65 (1H, m), 0.87-0.92 (1H, m), 0.97-1.04 (1H, m) , 1.11-1.18 (1H, m), 1.28-1.34 (1H, m), 1.64-1.71 (1H, m), 1.99-2.02 (1H, m), 2.45 (3H, s), 2.50 (3H, s), 2.71-2.76 (1H, m), 3.32-3.36 (3H, m), 3.64 -3.70 (1H, m), 4.01-4.05 (1H, m), 6.99 (1H, d, J = 9.06 Hz), 8.13 (1H, d, J = 9 , 06 Hz), 8.85 (1H, s).

Melting point: 190-192 ° C.

Elemental analysis for C ₂₂ H ₂₇ N ₃ O ₃ :

Calculated: C 69.27%; H 7.13%; N, 11.02%.

Found: C 69.00%; H 7.16%; N, 10.96%.

Reference Example 25

Ethyl 1- (2-bromoacetyl) cyclopropanecarboxylate

Ethyl 1-acetylcyclopropanecarboxylate (200 g, 1.28 mol) was dissolved in ethanol (1000 ml) and bromine (72.7 ml, 1.41 mol) was added dropwise with stirring and cooling on ice. After complete addition, the temperature of the reaction solution was raised to 30 ° C and stirring was carried out for 2 hours. After adding water (1000 ml) to the reaction solution while cooling on ice, it was concentrated under reduced pressure. After extracting the concentrate with ethyl acetate (750 ml × 2), it was washed with a 10% aqueous sodium thiosulfate solution (500 ml × 2) and saturated aqueous sodium bicarbonate (500 ml × 2), in that order, and then dried over anhydrous sodium sulfate. After filtering and concentrating the filtrate under reduced pressure, 291 g (97%) of the title compound was obtained as a yellow, oily substance.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.30 (3H, t, J = 7.1 Hz), 1.60-1.63 (4H, m), 4.22 (2H, q, J = 7.1 Hz), 4.49 (2H, s).

TLC: Rf = 0.7 (n-hexane: ethyl acetate = 3: 1).

Reference Example 26

Diethylphosphonoacetic acid

Ethyl diethylphosphonoacetate (100 g, 446 mmol) was dissolved in ethanol (275 ml) and after dropwise adding a 2 mol / L aqueous solution of sodium hydroxide (275 ml, 550 mmol), it was stirred at room temperature for 1 hour under ice-cooling. The reaction solution was then concentrated under reduced pressure, and the concentrate was acidified with concentrated hydrochloric acid while cooling on ice. It was extracted with ethyl acetate (200 ml × 4), chloroform (100 ml × 2) and 5% methanol / chloroform (250 ml × 2). The combined organic layers were then dried over anhydrous sodium sulfate and, after filtration, the filtrate was concentrated under reduced pressure. Thus, 89 g (quantitative yield) of the title compound was obtained as a colorless oily substance.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.35 (6H, t, J = 6.8 Hz), 2.98 (2H, d, J = 21.7 Hz), 4.19 (4H, q, J = 6.8 Hz).

TLC: Rf = 0.1 (chloroform: methanol = 9: 1).

Reference Example 27

Ethyl 1- [2- [N- [1- (S) -phenylethyl] amino] acetyl] cyclopropanecarboxylate

1- (S) -phenylethylamine (12.1 g, 100 mmol) was dissolved in acetonitrile (120 ml), an acetonitrile (50 ml) solution of triethylamine (15.3 ml, 110 mmol) was added dropwise and ethyl 1- was added dropwise. (2-Bromoacetyl) cyclopropanecarboxylate (23.5 g, 100 mmol) with stirring and cooling on ice. After complete addition, the reaction solution was stirred while cooling on ice for 1.5 hours. The reaction solution was then poured into water (75 ml) and concentrated under reduced pressure. The concentrate was extracted with diisopropyl ether (75 ml × 2) and then washed with water (75 ml). After extracting the organic layer with 1 mol / L hydrochloric acid (100 ml × 2), the acidic aqueous solution was washed with ethyl acetate (100 ml). After adding 1 mol / L aqueous sodium hydroxide solution (100 ml) to an acidic aqueous solution, and then adding further saturated aqueous sodium bicarbonate (100 ml), it was extracted with ethyl acetate (100 ml). The organic layer was then washed with water (100 ml) and brine (100 ml), in that order, and then dried over anhydrous sodium sulfate. After filtration and subsequent concentration of the filtrate under reduced pressure, 18.6 g (68%) of the title compound were obtained as a pale yellow oily substance.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.17 (3H, t, J = 7.1 Hz), 1.38 (3H, d, J = 6.6 Hz), 1.48 (4H, c), 3.71 (1H, q, J = 6.6 Hz), 3.86 (2H, d, J = 2.0 Hz), 4.10 (2H, q, J = 7.1 Hz) .

TLC: Rf = 0.6 (n-hexane: ethyl acetate = 1: 1).

Reference Example 28

Ethyl 1- [2- [N- (diethylphosphonoacetyl) -N- [1- (S) -phenylethyl] amino] acetyl] cyclopropanecarboxylate

Method A:

Diethylphosphonoacetic acid (15.1 g, 76.8 mmol) was dissolved in anhydrous tetrahydrofuran (120 ml) and, after addition of 1,1'-carbonyldiimidazole (13.7 g, 84.5 mmol), was stirred under ice cooling at room temperature in within 1 hour. After adding anhydrous tetrahydrofuran (30 ml) solution to the reaction solution, ethyl 1- [2- [N- [1- (S) -phenylethyl] amino] acetyl] cyclopropanecarboxylate (17.6 g, 64.0 mmol) under ice cooling it was stirred at room temperature for 1 hour. After adding 1 mol / L hydrochloric acid (100 ml) and ethyl acetate (100 ml) to the reaction solution, they were extracted and the organic layer was separated. The aqueous layer was then extracted with ethyl acetate (100 ml) and the combined organic layer was washed with saturated aqueous sodium bicarbonate (100 ml) and saturated brine (100 ml), in that order, and then dried over anhydrous sodium sulfate. After filtration and subsequent concentration of the filtrate under reduced pressure, 28.7 g (99%) of the title compound was obtained as a yellow syrup.

Method B

Diethylphosphonoacetic acid (32.8 g, 166 mmol) was dissolved in anhydrous benzene (700 ml) and after adding N, N'-dimethylformamide (1 ml), thionyl chloride (18.2 ml, 250 mmol) was added and refluxed for 1.5 hours. Then the reaction solution was allowed to cool, concentrated under reduced pressure, and after adding dried toluene (100 ml), it was again concentrated under reduced pressure. After repeating this operation 3 times, the concentrate was dissolved in anhydrous tetrahydrofuran (300 ml), and after adding anhydrous tetrahydrofuran (300 ml) solution of ethyl 1- [2- [N- [1- (S) -phenylethyl] amino] acetyl] cyclopropanecarboxylate (45.7 g, 166 mmol) and triethylamine (25.1 ml, 183 mmol) with stirring and cooling on ice, it was stirred while cooling on ice for 1.5 hours and then stirred at room temperature for 2 hours. After adding 1 mol / L hydrochloric acid (300 ml) and ethyl acetate (300 ml) to the reaction solution, and the extraction was performed, the organic layer was separated. The aqueous layer was then extracted with ethyl acetate (300 ml), and the combined organic layer was washed with saturated aqueous sodium bicarbonate (300 ml) and saturated brine (300 ml), in that order, and then dried over anhydrous sodium sulfate. After filtration and subsequent concentration of the filtrate under reduced pressure, 43.6 g (70%) of the title compound was obtained as a yellow syrup.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.14, 1.20 (3H, t, J = 7.1 Hz), 1.29-1.68 (13H, m), 2.85, 4 69 (2H, dd, J = 9.5, 20.7 Hz), 3.18, 4.55 (2H, d, J = 22.2 Hz), 4.06-4.22 (6H, m ), 5.42, 6.05 (1H, q, J = 7.1 Hz), 7.26-7.37 (5H, m).

MS (m / z): 454 ([M + H]) ⁺ .

TLC: Rf = 0.1 (n-hexane: ethyl acetate = 1: 1).

Reference Example 29

4- (1-ethoxycarbonylcyclopropyl) -1- [1- (S) -phenylethyl] -3-pyrrolin-2-one

After dissolving ethyl 1- [2- [N- (diethylphosphonoacetyl) -N- [1- (S) -phenylethyl] amino] acetyl] cyclopropanecarboxylate (25.0 g, 55.2 mmol) in toluene (250 ml) was gradually added potassium tert-butoxide (7.40 g, 66.2 mmol) with stirring and cooling on ice. After stirring the reaction solution for 15 minutes at room temperature, a 10% aqueous solution of citric acid (250 ml) and ethyl acetate (250 ml) were added, and after the extraction, the organic layer was separated. The aqueous layer was then extracted with ethyl acetate (250 ml), and after washing the combined organic layer with saturated aqueous sodium bicarbonate (250 ml) and saturated brine (250 ml), in that order, the organic layer was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, the concentrate was applied to silica gel for chromatography, and 12.1 g (73%) of the title compound was obtained as an orange syrup from n-hexane: ethyl acetate = 2: 1-1: 2 eluate. Instrumental analysis data of the obtained compound are consistent with the data given in PCT / JP96 / 00208.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.18, (3H, t, J = 7.2 Hz), 1.60-1.63 (7H, m), 3.80 (1H, dd, J = 1.5, 9.0 Hz), 4.07-4.11 (2H, m), 4.13 (1H, d, J = 9.0 Hz), 5.55 (1H, q, J = 7.1 Hz), 5.84 (1H, t, J = 1.5 Hz), 7.24-7.36 (5H, m).

MS (m / z): 300 ([M + H]) ⁺ .

TLC: Rf = 0.5 (n-hexane: ethyl acetate = 1: 1).

Reference Example 30

4- (S) - (1-ethoxycarbonylcyclopropyl) -1- [1- (S) -phenylethyl] pyrrolidin-2-one

4- (1-ethoxycarbonylcyclopropyl) -1- [1- (S) -phenylethyl] -3-pyrrolin-2-one (12.1 g, 40.5 mmol) was dissolved in ethyl acetate (120 ml), 5% catalyst was added palladium on carbon (water content: 50%, 2.4 g) and stirred at room temperature for 17 hours in a hydrogen atmosphere at atmospheric pressure. The reaction solution was then filtered through celite (washed with ethyl acetate) and the filtrate was then concentrated under reduced pressure. The concentrate was applied to silica gel for chromatography, and from n-hexane: ethyl acetate = 3: 2 eluate, 9.00 g (74%) of the title compound was obtained as a pale yellow syrup. In addition, 2.60 g (21%) of the diastereomer of the (4- (R) -isomer) of the title compound was obtained as a pale yellow syrup. Instrumental analysis data of the obtained compound are consistent with the data given in PCT / JP96 / 00208.

4- (S) -isomer:

¹ H NMR (400 MHz, CDCl ₃ ) δ: 0.63-0.65 (2H, m), 1.13 (3H, t, J = 7.1 Hz), 1.12-1.19 (2H , m), 1.52 (3H, d, J = 7.3 Hz), 2.17 (1H, dd, J = 9.0, 16.8 Hz), 2.46 (1H, dd, J = 9.3, 16.3 Hz), 2.67-2.76 (2H, m), 3.47 (1H, t, J = 8.3 Hz), 3.96-4.11 (2H, m ), 5.51 (1H, q, J = 7.3 Hz), 7.26-7.35 (5H, m).

TLC: Rf = 0.45 (n-hexane: ethyl acetate = 1: 1).

4- (R) -isomer:

¹ H NMR (400 MHz, CDCl ₃ ) δ: 0.72-0.76 (2H, m), 1.18-1.24 (2H, m), 1.21 (3H, t, J = 7, 1 Hz), 1.52 (3H, d, J = 7.1 Hz), 2.27-2.32 (1H, m), 2.44-2.52 (2H, m), 3.14 ( 2H, d, J = 8.0 Hz), 4.10 (2H, q, J = 7.1 Hz).

TLC: Rf = 0.5 (n-hexane: ethyl acetate = 1: 1).

Reference Example 31

1- [1- [1- (S) -phenylethyl] -2-one-4- (S) -pyrrolidin-4-yl] -1-cyclopropanecarboxylic acid

4- (S) - (1-ethoxycarbonylcyclopropyl) -1- [1- (S) -phenylethyl] pyrrolidin-2-one (10.5 g, 34.9 mmol) was dissolved in 70 ml of ethanol, and after adding 1 mol / l of an aqueous solution of sodium hydroxide (70 ml) while cooling on ice, the reaction solution was stirred at room temperature for 15.5 hours and then at 40 ° C for 3 hours. After concentrating the reaction solution under reduced pressure, the residual aqueous layer was washed with ethyl acetate (70 ml). The aqueous layer was then acidified with concentrated hydrochloric acid while cooling on ice and then extracted with chloroform (70 ml × 3). The organic layer was then dried over anhydrous sodium sulfate, and after filtration, the filtrate was concentrated under reduced pressure. Thus, 9.40 g (99%) of the title compound was obtained as a white solid.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 0.72-0.74 (2H, m), 1.21-1.23 (2H, m), 1.52 (3H, d, J = 7, 3 Hz), 2.17 (1H, dd, J = 8.8, 16.8 Hz), 2.48 (1H, dd, J = 9.5, 16.8 Hz), 2.66-2, 78 (2H, m), 3.50 (1H, t, J = 9.3 Hz), 5.51 (1H, q, J = 7.3 Hz), 7.25-7.34 (5H, m )

Reference Example 32

4- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] -1- [1- (S) -phenylethyl] pyrrolidin-2-one

A toluene (15 ml) solution of triethylamine (9.6 ml, 69 mmol) and diphenylphosphoric azide (DPPA; 10.4 g, 37.9 mmol) were added to the toluene (80 ml) suspension of 1- [1- [1- ( S) -phenylethyl] -2-one-4- (S) -pyrrolidin-4-yl] -1-cyclopropanecarboxylic acid (9.4 g, 34.4 mmol), and after stirring at room temperature for 1 hour in nitrogen atmosphere was refluxed for 1.5 hours. After the reaction solution was cooled to room temperature, tert-butyl alcohol (95 ml) was added and refluxed for 15 hours. Then the reaction solution was allowed to cool, the reaction solution was concentrated under reduced pressure, and ethyl acetate (95 ml) and water (95 ml) were added to the concentrate. After extraction, the organic layer was separated and the aqueous layer was extracted with ethyl acetate (95 ml). The combined organic layer was then washed with brine (95 ml) and then dried over anhydrous sodium sulfate. After filtering and concentrating the filtrate under reduced pressure, the concentrate was applied to silica gel for chromatography and 10.7 g (90%) of the title compound was obtained from the chlorine: methanol = 50: 1 eluate as a colorless, amorphous substance.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 0.56-0.85 (4H, m), 1.37 (9H, s), 1.51 (3H, d, J = 7.3 Hz), 2.32-2.44 (3H, m), 2.79 (1H, dd, J = 7.3, 10.0 Hz), 3.36 (1H, m), 4.66 (1H, br. c), 5.50 (1H, q, J = 7.3 Hz), 7.26-7.34 (5H, m).

TLC: Rf = 0.15 (n-hexane: ethyl acetate = 1: 1).

Reference Example 33

3- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] -1- [1- (S) -phenylethyl] pyrrolidine

4- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] -1- [1- (S) -phenylethyl] pyrrolidin-2-one (10.4 g, 30.2 mmol) was dissolved in anhydrous tetrahydrofuran (100 ml) and a 1.0 M solution of boron-tetrahydrofuran / tetrahydrofuran complex (90.7 ml, 90.7 mmol) was added dropwise in a nitrogen atmosphere under ice-cooling. After completion of the addition, it was stirred for 16 hours at a temperature ranging from cooling on ice to room temperature. After slowly adding an aqueous solution (100 ml) of potassium carbonate (25.0 g, 181 mmol) to the reaction solution while cooling on ice, it was heated under reflux for 1.5 hours. The reaction solution was then allowed to cool, extracted with ethyl acetate (100 ml × 2), and then washed with brine (100 ml). The organic layer was then dried over anhydrous sodium sulfate, and after filtration, the filtrate was concentrated under reduced pressure. The concentrate was applied to silica gel for chromatography, and 8.20 g (82%) of the title compound were obtained as colorless crystals from chloroform: methanol = 100: 1-30: 1 eluate.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 0.62 (2H, br s), 0.75-0.88 (2H, m), 1.35 (3H, d, J = 6.6 Hz ), 1.41 (9H, s), 1.63 (2H, m), 1.88-1.92 (1H, m), 2.14-2.17 (1H, m), 2.27- 2.34 (2H, m), 2.63 (1H, br s), 3.15 (1H, similar t, J = 6.6 Hz), 5.10 (1H, br s), 7, 23-7.33 (5H, m).

MS (m / z): 331 ([M + H]) ⁺ .

TLC: Rf = 0.4 (chloroform: methanol = 9: 1).

Reference Example 34

3- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] pyrrolidine

3- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] -1- [1- (S) -phenylethyl] pyrrolidine (270 mg, 0.817 mmol) was dissolved in ethanol (15 ml), and after adding the catalyst 10% palladium on carbon (water content: 52.0%; 270 mg) was stirred at 40 ° C for 3 hours in a hydrogen atmosphere at atmospheric pressure. After removal of the catalyst by filtration (washing with ethanol), the filtrate was concentrated under reduced pressure to obtain 185 mg (quantitative yield) of the title compound as a colorless syrup.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 0.69 (2H, broad s), 0.79 (2H, broad s), 1.42 (9H, s), 1.43-1.50 (1H, m), 1.86-1.88 (1H, m), 2.15-2.19 (1H, m), 2.68-2.72 (1H, m), 2.90-3 07 (3H, m); 4.92 (1H, broad s).

Reference Example 35

1-Benzyloxycarbonyl-3- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] pyrrolidine

3- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] -1- [1- (S) -phenylethyl] pyrrolidine (1.70 g, 5.15 mmol) was dissolved in dichloromethane (34 ml) and dropwise benzyl chloroformate (1.10 ml, 7.73 mmol) was added with stirring and cooling on ice. The reaction solution was stirred at room temperature for 2.5 hours and then stirred at 40 ° C for 1.5 hours. Then the reaction solution was allowed to cool, the reaction solution was concentrated under reduced pressure, the concentrate was applied to silica gel for chromatography, and 1.40 g (75%) of the title compound was obtained as a colorless syrup from n-hexane: ethyl acetate = 2: 1 eluate . Instrumental analysis data of the obtained compound are consistent with the data given in PCT / JP96 / 00208.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 0.70 (2H, broad s), 0.80 (2H, broad s), 1.41 (9H, s), 1.63 (1H, m ), 1.92 (1H, m), 2.25 (1H, m), 3.07-3.12 (1H, m), 3.29-3.31 (1H, m), 3.56 ( 2H, m), 4.85 (1H, broad s), 5.12 (2H, s), 7.33-7.36 (5H, m).

TLC: Rf = 0.4 (n-hexane: ethyl acetate = 1: 1).

Reference Example 36

1-Benzyloxycarbonyl-3- (R) - [1- [N- (tert-butoxycarbonyl) -

N- (methyl) amino] cyclopropyl] pyrrolidine

1-Benzyloxycarbonyl-3- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] pyrrolidine (1.40 g, 3.89 mmol), N, N'-dimethylformamide (7 ml), silver oxide (9.0 g, 39 mmol) and methyl iodide (24 ml, 389 mmol) were placed in a darkened and sealed tube, and the mixture was stirred for 13 hours in an oil bath at 80 ° C. The reaction solution was then filtered through celite (washed with ethyl acetate), and the filtrate was then diluted with ethyl acetate (100 ml). The organic layer was then washed with water (50 ml × 3) and brine (50 ml), in that order, and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, the concentrate was applied to silica gel for chromatography, and 1.31 g (89%) of the title compound was obtained as a pale yellow syrup from n-hexane: ethyl acetate = 2: 1 eluate. Instrumental analysis data of the obtained compound are consistent with the data given in PCT / JP96 / 00208.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 0.83 (4H, br s), 1.42 (9H, s), 1.55 (1H, m), 1.88 (1H, m), 2.28-2.43 (1H, m), 2.83 (3H, s), 3.02-3.04 (1H, m), 3.25-3.33 (1H, m), 3, 55 (2H, m), 5.12 (2H, s), 7.32-7.35 (5H, m).

MS (m / Z): 374 ([M + H]) ⁺ .

TLC: Rf = 0.4 (n-hexane: ethyl acetate = 1: 1).

Reference Example 37

3- (R) - [1- [N- (tert-butoxycarbonyl) -N- (methyl) amino] cyclopropyl] pyrrolidine

1-benzyloxycarbonyl-3- (R) - [1- [N- (tert-butoxycarbonyl) -N- (methyl) amino] cyclopropyl] pyrrolidine (1.31 g, 3.48 mmol) was dissolved in ethanol (13 ml) , and after the addition of the catalyst, 10% palladium on carbon (water content: 50%; 0.65 g) was stirred at room temperature for 5 hours in a hydrogen atmosphere at atmospheric pressure. After filtering through celite (washing with ethanol), the filtrate was concentrated under reduced pressure. The result was 0.92 g (quantitative yield) of the title compound as a colorless syrup.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 0.80 (4H, broad s), 1.46 (9H, s), 1.81 (1H, m), 2.04 (1H, broad c ), 2.28-2.42 (1H, m), 2.54 (1H, br s), 2.84 (3H, s), 2.88-2.96 (3H, m).

TLC: Rf = 0.1 (chloroform: methanol = 9: 1).

Reference Example 38

3- (R) - [1- [N- (tert-butoxycarbonyl) -N- (methyl) amino] cyclopropyl] -1- [1- (S) -phenylethyl] pyrrolidin-2-one

4- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] -1- [1- (S) -phenylethyl] pyrrolidin-2-one (7.87 g, 22.8 mmol) was dissolved in dimethylformamide (100 ml ) while cooling on ice, 60% sodium hydride in oil (1.10 g, 27.4 mmol) was added and stirred for 5 minutes. Then, methyl iodide (7.11 ml, 114 mmol) was added dropwise with stirring at room temperature. After complete addition, the reaction suspension was stirred at room temperature for 14 hours. After adding 60% sodium hydride in oil (296 mg, 7.40 mmol) and methyl iodide (1.00 ml, 16.1 mmol), the suspension was stirred at 40 ° C. for 24 hours. After a saturated aqueous solution of ammonium chloride (100 ml) and water (150 ml) were added to the reaction suspension, they were extracted with ethyl acetate (300 ml × 2) with stirring and cooling on ice. The combined organic layer was then washed with water (100 ml × 2) and brine (100 ml × 2), in that order, and then dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, the obtained residue was applied to silica gel for chromatography, and chloroform: methanol = 50-30: 1 gave 7.53 g (92%) of the title compound as a colorless oily substance from the eluate.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 0.83 (4H, m), 1.32 (6H, s), 1.38 (3H, s), 1.51 (3H, d, J = 7 , 1 Hz), 1.61 (3H, d, J = 16.6 Hz), 2.43 (1H, m), 2.68-2.81 (3H, m), 3.21 (1H, m ), 5.48-5.50 (1H, m), 7.26-7.36 (5H, m).

Reference Example 39

3- (R) - [1- [N- (tert-butoxycarbonyl) -N- (methyl) amino] cyclopropyl] -1- [1- (S) -phenylethyl] pyrrolidine

3- (R) - [1- [N- (tert-butoxycarbonyl) -N- (methyl) amino] cyclopropyl] -1- [1- (S) -phenylethyl] pyrrolidin-2-one (7.53 g, 21.0 mmol) was dissolved in anhydrous tetrahydrofuran (70 ml), and a 1.0 M solution of the boron tetrahydrofuran / tetrahydrofuran complex (63.0 ml, 63.0 mmol) was gradually added dropwise with stirring and cooling on ice. After complete addition, the solution was stirred at room temperature for 20 hours. After slowly adding an aqueous solution (72 ml) of potassium carbonate (7.22 g) while cooling on ice, the mixture was refluxed for 1.5 hours. The reaction solution was then allowed to cool to room temperature, water (150 ml) was added, extracted with ethyl acetate (200 ml × 2), then the combined organic layer was washed with saturated hydrochloric solution (200 ml) and then dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, the concentrate was applied to silica gel for chromatography, and chlorine form: methanol = 50: 1 gave 7.19 g (99%) of the title compound as a colorless syrup from the eluate.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 0.73 (4H, m), 1.35 (9H, s), 1.36 (3H, s), 1.61 (1H, m), 1, 85 (1H, m), 1.97 (1H, m), 2.27 (1H, m), 2.50-2.58 (2H, m), 2.79 (3H, s), 2.99 (1H, m), 3.14-3.19 (1H, m), 7.27-7.30 (5H, m).

Reference Example 40

3- (R) - [1- [N- (tert-butoxycarbonyl) -

N- (methyl) amino] cyclopropyl] pyrrolidine

3- (R) - [1- [N- (tert-butoxycarbonyl) -N- (methyl) amino] cyclopropyl] -1- [1- (S) -phenylethyl] pyrrolidine (7.19 g, 20.9 mmol ) was dissolved in ethanol (78 ml) and, after adding a catalyst, 10% palladium on carbon (water content: 50%; 3.9 g) was stirred at 40 ° C for 4 hours in a hydrogen atmosphere at atmospheric pressure. After filtering through celite (washing with ethanol), the filtrate was concentrated under reduced pressure. The result was 4.38 g (quantitative yield) of the title compound as a colorless syrup. ¹ H-NMR data and Rf TLC of the obtained compound are consistent with the data shown before.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 0.80 (4H, broad s), 1.46 (9H, s), 1.81 (1H, m), 2.04 (1H, broad c ), 2.28-2.42 (1H, m), 2.54 (1H, br s), 2.84 (3H, s), 2.88-2.96 (3H, m).

TLC: Rf = 0.1 (chloroform: methanol = 9: 1).

Reference Example 41

4- (R) - [1- [N- (tert-butoxycarbonyl) -N- (ethyl) amino] cyclopropyl] -1- [1- (S) -phenylethyl] pyrrolidin-2-one

4- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] -1- [1- (S) -phenylethyl] pyrrolidin-2-one (4.16 g, 12.1 mmol) was dissolved in dimethylformamide (50 ml ) 60% sodium hydride in oil (580 mg, 14.5 mmol) was added to the mixture under a nitrogen atmosphere and at room temperature, stirred for 10 minutes and then ethyl iodide (4.87 ml, 60.5 mmol) was added dropwise. After complete addition, the reaction suspension was stirred at room temperature for 15 hours. After a saturated aqueous solution of ammonium chloride (150 ml) was added to the reaction suspension, it was extracted with ethyl acetate (150 ml × 2) with stirring and cooling on ice. The combined organic layer was then washed with water (150 ml × 2) and brine (150 ml), in that order, and then dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, the obtained residue was applied to silica gel for chromatography, and 4.56 g (quantitative yield) of the title compound was obtained as a colorless oily substance from the n-hexane: ethyl acetate = 1: 2 eluate.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 0.49-0.80 (4H, m), 1.02-1.04 (3H, m), 1.37 (9H, s), 1.49 -1.51 (3H, m), 1.92-1.94 (1H, m), 2.04-2.06 (1H, m), 2.36-2.38 (1H, m), 2 , 67-2.70 (2H, m), 3.20-4.23 (2H, m), 5.48-5.50 (1H, m), 7.26-7.52 (5H, m) .

Reference Example 42

3- (R) - [1- [N- (tert-butoxycarbonyl) -N- (ethyl) amino] cyclopropyl] -1- [1- (S) -phenylethyl] pyrrolidine

4- (R) - [1- [N- (tert-butoxycarbonyl) -N- (ethyl) amino] cyclopropyl] -1- [1- (S) -phenylethyl] pyrrolidin-2-one (4.56 g, 12.1 mmol) was dissolved in anhydrous tetrahydrofuran (80 ml), and a 1.0 M solution of boron tetrahydrofuran / tetrahydrofuran complex (48.0 ml, 48.0 mmol) was added dropwise with stirring and cooling on ice. After complete addition, the reaction solution was stirred for 16 hours under conditions of cooling on ice to room temperature. After concentrating the reaction solution under reduced pressure, a mixed solution of 9: 1 (100 ml) ethanol and water was added, and after the addition of triethylamine (5 ml), it was heated under reflux for 4 hours. After the reaction solution was cooled to room temperature, it was concentrated under reduced pressure, a saturated sodium carbonate aqueous solution (100 ml) was added to the residue, extracted with chloroform (100 ml × 2), then the combined organic layer was washed with brine (100 ml) and then dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, the concentrate was applied to silica gel for chromatography, and chloroform: methanol = 100: 1-95: 5 eluate gave 4.26 g (99%) of the title compound as a colorless syrup.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 0.54-0.78 (4H, m), 1.09-1.11 (3H, m), 1.33-1.43 (13H, m) 1.84-1.97 (2H, m), 2.26-2.28 (2H, m), 2.56-2.57 (2H, m), 2.86-2.96 (1H, m), 3.13-3.18 (2H, m), 7.21-7.30 (5H, m).

Reference Example 43

3- (R) - [1- [N- (tert-butoxycarbonyl) -N- (ethyl) amino] cyclopropyl] pyrrolidine

3- (R) - [1- [N- (tert-butoxycarbonyl) -N- (ethyl) amino] cyclopropyl] -1- [1- (S) -phenylethyl] pyrrolidine (3.01 g, 8.40 mmol ) was dissolved in ethanol (120 ml) and after adding a catalyst 10% palladium on carbon (water content: 50%; 3.0 g) was stirred at 40 ° C for 5 hours in a hydrogen atmosphere at atmospheric pressure. After filtration through celite (washed with ethanol), the filtrate was concentrated under reduced pressure. The result was 2.16 g (quantitative yield) of the title compound as a colorless syrup.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 0.83-0.85 (4H, m), 1.46 (9H, s), 1.74-1.82 (1H, m), 2.16 -2.18 (2H, m), 2.43-2.52 (2H, m), 2.90-2.99 (2H, m), 3.21-3.24 (2H, m).

Reference Example 44

2,3,4-Trichlorobenzoic acid

Sodium hydroxide (45.42 g, 1.090 mol) was dissolved in water (220 ml) and bromine (16.85 ml, 0.327 mol) was added dropwise with cooling on ice over 5 minutes. After stirring the reaction solution at 0 ° C for 15 minutes, a solution of dioxane (220 ml) of 2 ', 3', 4'-trichloroacetophenone (24.40 g, 0.109 mol) was added dropwise at 0 ° C over 30 minutes. After stirring at room temperature for 14 hours, water (350 ml) was added and then washed with dichloromethane (350 ml). The resulting aqueous layer was gradually acidified with concentrated hydrochloric acid while cooling on ice, and the resulting crystals were filtered. After washing the filtered crystals with water, water was removed by azeotropic distillation with toluene. Thus, 22.33 g (91%) of the title compound was obtained as a pale yellow powder.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.58 (1H, d, J = 8.3 Hz), 7.70 (1H, d, J = 8.6 Hz).

Reference Example 45

Ethyl (2,3,4-trichlorobenzoyl) acetate

2,3,4-Trichlorobenzoic acid (4.51 g, 20.0 mmol) was dissolved in tetrahydrofuran (80 ml), carbonyldiimidazole (3.89 g, 24.0 mmol) was added under ice-cooling, and then stirred at room temperature within 3 hours (solution A). Meanwhile, the monopotassium salt of malonic acid monoethyl ester (6.81 g, 40.0 mmol) was suspended in ethyl acetate and triethylamine (13.9 ml, 100.0 mmol) and magnesium chloride (5.71 g, 60.0 mmol). After stirring at room temperature for 3 hours, the reaction solution was cooled on ice, and the above solution A was added dropwise to this reaction solution over 10 minutes. After washing solution A in the reaction solution using tetrahydrofuran (10 ml), it was stirred at room temperature for 14 hours, and then the reaction solution was poured into a 10% aqueous citric acid solution (200 ml). The mixture was then extracted with ethyl acetate (200 ml), washed with saturated hydrochloric solution (200 ml) and then dried over anhydrous sodium sulfate. Then, after removing the drying agent by filtration, the solvent was evaporated under reduced pressure, and the obtained crude product was purified by silica gel chromatography, thereby obtaining n-hexane: ethyl acetate = 5: 1, 2.681 g (45%) of the title compound as a pale -red oil.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.25 (1.5H, t, J = 7.2 Hz), 1.34 (1.5H, t, J = 7.0 Hz), 3, 99 (1H, s) 4.19 (1H, q, J = 7.2 Hz), 4.28 (1H, q, J = 7.0 Hz), 5.47 (0.5H, s), 7 37-7.49 (2H, m); 12.45 (0.5H, m).

Reference Example 46

Ethyl 7,8-dichloro-1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carboxylate

A mixture of ethyl (2,3,4-trichlorobenzoyl) acetate (2.681 g, 9.07 mmol), acetic anhydride (10 ml) and triethyl orthoformate (20 ml) was refluxed for 2.5 hours in an oil bath at 140 ° C. After the solvent was evaporated under reduced pressure, azeotropic distillation was performed using toluene (3 times) to obtain 3.272 g of crude ethyl 3-ethoxy-2- (2,3,4-trichlorobenzoyl) acrylate as a pale red oil.

The crude product of ethyl 3-ethoxy-2- (2,3,4-trichlorobenzoyl) acrylate obtained above (3.272 g) was dissolved in dichloromethane (50 ml), 2- (S) -fluoro-1- (R) -cyclopropylamine tosylate was added. (2.467 g, 9.98 mmol) and triethylamine (1.64 ml, 11.79 mmol), in that order, while cooling on ice soaked in salt, and stirred at room temperature for 19.5 hours. Ethyl acetate (200 ml) was added to the reaction solution, and after washing with a 10% aqueous citric acid solution (80 ml × 2), saturated aqueous sodium bicarbonate (80 ml) and saturated brine (80 ml), dried over anhydrous sodium sulfate. Then, after removing the drying agent by filtration, the solvent was evaporated under reduced pressure to obtain crude ethyl 3- [2- (S)-fluoro-1- (R) -cyclopropyl] amino-2- (2,3,4-trichlorobenzoyl) acrylate (3.59 g) as a pale orange, gummy substance.

The above crude ethyl 3- [2- (S) -fluoro-1- (R) -cyclopropyl] amino-2- (2,3,4-trichlorobenzoyl) acrylate (3.57 g) was dissolved in dried dioxane (45 ml ) and after adding sodium hydride there (60%, 433 mg, 10.82 mmol) while cooling on ice, it was stirred for 14 hours while heating in an oil bath at 50 ° C. After evaporating the solvent under reduced pressure, the residue was dissolved in chloroform (150 ml), and after washing with a 10% aqueous citric acid solution (50 ml) and brine (50 ml), dried over anhydrous sodium sulfate. Then, after removing the drying agent by filtration, the solvent was evaporated under reduced pressure, and the obtained crude product was purified by silica gel chromatography, thereby obtaining chloroform: ethyl acetate = 1: 2, 1.475 g (48%) of the title compound as pale yellow from the eluate powder.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.35-1.50 (1H, m), 1.41 (3H, t, J = 7.1 Hz), 1.55-1.75 (1H , m), 4.08-4.13 (1H, m), 4.39 (2H, q, J = 7.1 Hz), 4.80-4.98 (1H, m), 7.53 ( 1H, d, J = 8.5 Hz), 8.34 (1H, d, J = 6.8 Hz), 8.57 (1H, d, J = 2.7 Hz).

MS (m / z): 344 (M ⁺ ).

Reference Example 47

7,8-dichloro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carboxylic acid

A mixture of ethyl 7,8-dichloro-1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carboxylate (1.114 g, 3.237 mmol), acetic acid (8 ml) and concentrated hydrochloric acid (4 ml) were refluxed for 2 hours in an oil bath at 130 ° C. Then, after adding water (40 ml) and cooling on ice, the obtained crystals were filtered and washed with water (5 ml × 2), 5% aqueous ethanol (5 ml × 2) and diethyl ether (5 ml × 2), thereby obtaining 909 mg (89%) of the title compound as a pale yellow powder.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.40-1.80 (2H, m), 4.23-4.28 (1H, m), 4.83-5.02 (1H, m) 7.67 (1H, d, J = 8.8 Hz), 8.38 (1H, d, J = 8.6 Hz), 8.86 (1H, d, J = 2.7 Hz).

Melting point: 198-201 ° C.

Specific rotation: [α] _D ^24.5 = -24.0 °.

Elemental analysis for C ₁₃ H ₈ Cl ₂ FNO ₃ :

Calculated: C 49.39%; H 2.55%; N, 4.43%.

Found: C 49.14%; H 2.40%; N, 4.33%.

MS (m / z): 315 (M ⁺ ), 354 [(M + K) ⁺ ].

EXAMPLE 16

7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -8-chloro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro -4-oxoquinoline-3-carboxylic acid

Mixture of 7.8-dichloro-1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carboxylic acid (253 mg, 0.80 mmol) , 3- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] pyrrolidine (272 mg, 1.20 mmol), N-methylpiperidine (0.195 ml, 1.60 mmol) and dimethyl sulfoxide (3 ml) were stirred for 55 hours when heated in an oil bath at 80 ° C and in a nitrogen-substituted atmosphere. After evaporation of the solvent, the residue was partitioned between ethyl acetate (50 ml) and 10% aqueous citric acid solution (30 ml), and after the organic layer was separated, the organic layer was washed with saturated hydrochloric solution (30 ml). The organic layer thus obtained was dried over anhydrous sodium sulfate, and after removing the drying agent by filtration, the solvent was evaporated under reduced pressure. The residue was purified by silica gel chromatography and from chloroform: methanol = 10: 1 eluate, crude 7- {3- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] pyrrolidin-1-yl} -8-chloro-1- [ 2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carboxylic acid.

The above crude 7- {3- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] pyrrolidin-1-yl} -8-chloro-1- [2- (S) -fluoro-1- (R) - cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carboxylic acid was dissolved in concentrated hydrochloric acid (5 ml) under ice cooling, and after stirring at room temperature for 20 minutes, the solution was poured into a separatory funnel and washed chloroform (10 ml × 10 times or more). After washing, a saturated aqueous solution of sodium hydroxide was added to the aqueous layer while cooling on ice, thereby bringing the pH to> 11, and then the pH was adjusted to 7.7 by the addition of concentrated hydrochloric acid and 1 mol / L hydrochloric acid. The resulting aqueous layer was then extracted with chloroform (100 ml) and a mixture of chloroform: methanol = 9: 1 (100 ml × 2) and the combined organic layer was dried over anhydrous sodium sulfate. After removing the drying agent by filtration, the solvent was evaporated under reduced pressure. The residue was then purified by preparative chromatography (separation in the lower layer with chloroform: methanol: water, 7: 3: 1), purification of the suspension using ethanol-diethyl ether, and then dried under reduced pressure to obtain 96 mg (30%) of the title compounds in the form of a pale yellow powder.

¹ H NMR (400 MHz, 0.1 mol / L NaOD / D ₂ O) δ: 0.45-0.65 (4H, m), 1.15-1.30 (1H, m), 1.50 -1.75 (2H, m), 2.00-2.10 (1H, m), 2.10-2.25 (1H, m), 3.25-3.40 (2H, m), 3 55-3.75 (2H, m), 4.10-4.15 (1H, m), 4.90-5.15 (1H, m), 7.05 (1H, d, J = 9, 0 Hz), 7.99 (1H, d, J = 9.3 Hz), 8.39 (1H, d, J = 3.7 Hz).

Melting point: 128-130 ° C.

Specific rotation: [α] _D ^24.5 = -193.0 °.

Elemental analysis for C ₂₀ H ₂₁ ClFN ₃ O ₃ · 1.5 H ₂ O:

Calculated: C 55.49%; H 5.59%; N, 9.71%.

Found: C 55.74%; H 5.45%; N, 9.57%.

MS (m / z): 406 [(M + H) ⁺ ].

Reference Example 48

Ethyl 7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -5-hydroxy-8-methyl-1,4-dihydro-4-oxoquinoline-3-carboxylate

Ethyl 5-amino-7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -8-methyl-1,4-dihydro-4-oxoquinoline-3-carboxylate (1.414 g, 4.39 mmol) was suspended in a 35% aqueous solution of sulfuric acid (15 ml), and an aqueous solution of sodium nitrite (394 mg, 5.70 mmol / 4.5 ml) was added dropwise under ice-cooling over 5 minutes. Then, after stirring at 0 ° C for 30 minutes, a small amount of urea was added and at the same temperature an aqueous solution of copper (II) nitrate trihydrate (17.00, 70.2 mmol / 155 ml) was added dropwise. After stirring at 0 ° C for 5 minutes, copper (I) oxide (565 mg, 3.95 mmol) was added with vigorous stirring of the reaction solution. Then, after stirring at room temperature for 20 minutes, extraction was performed with chloroform (200 ml × 2), and after slightly alkalizing the aqueous layer with sodium bicarbonate, the aqueous layer was extracted with chloroform (150 ml × 3). The combined organic layer was then dried over anhydrous sodium sulfate, and after removing the drying agent by filtration, the solvent was evaporated under reduced pressure. The resulting crude product was purified by silica gel chromatography, thereby obtaining chloroform: methanol = 10: 1 297 mg (21%) of the title compound as a yellow powder from the eluate.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.36-1.47 (1H, m), 1.40 (3H, t, J = 7.1 Hz), 1.53-1.63 (1H , m), 2.51 (3H, t, J = 2.4 Hz), 3.85-3.90 (1H, m), 4.39 (2H, q, J = 7.2 Hz), 4 77-4.96 (1H, m); 6.58 (1H, d, J = 11.5 Hz); 8.52 (1H, d, J = 3.2 Hz).

MS (m / z): 324 [(M + H) ⁺ ].

Reference Example 49

7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -5-hydroxy-8-methyl-1,4-dihydro-4-oxoquinoline-3-carboxylic acid

Mixture of ethyl 7-fluoro-1- [2- (S) -fluoro-1- (R) -cyclopropyl] -5-hydroxy-8-methyl-1,4-dihydro-4-oxoquinoline-3-carboxylate (325 mg , 1.005 mmol), acetic acid (3 ml) and concentrated hydrochloric acid (1.5 ml) were refluxed for 2 hours in an oil bath at 120 ° C. After adding water (30 ml) and cooling on ice, the obtained crystals were filtered off and then washed with water, 5% aqueous ethanol and diethyl ether to obtain 267 mg (90%) of the title compound as a pale yellow powder.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.40-1.75 (2H, m), 2.58 (3H, t, J = 2.5 Hz), 3.99-4.04 (1H , m), 4.80-5.05 (1H, m), 6.70 (1H, d, J = 11.3 Hz), 8.76 (1H, d, J = 3.2 Hz), 13 17 (0.7H, d, J = 1.0 Hz); 13.34 (0.7H, br s).

Melting point: 209-213 ° C.

Specific rotation: [α] _D ^24.7 = -111.6 °.

Elemental analysis for C ₁₄ H ₁₁ F ₂ NO ₄ :

Calculated: C 56.95%; H 3.76%; N 4.74%.

Found: C 56.90%; H 3.74%; N 4.68%.

MS (m / z): 296 [(M + H) ⁺ ].

EXAMPLE 17

7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -5-hydroxy-8-methyl-1 , 4-dihydro-4-oxoquinoline-3-carboxylic acid

Mixture of 7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -5-hydroxy-8-methyl-1,4-dihydro-4-oxoquinoline-3-carboxylic acid (205 mg 0.694 mmol), 3- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] pyrrolidine (314 mg, 1.388 mmol), N-methylpiperidine (0.243 ml, 1.388 mmol) and dimethyl sulfoxide (1.5 ml) were mixed in for 66 hours when heated in an oil bath at 80 ° C in a nitrogen-substituted atmosphere. After evaporation of the solvent, the residue was partitioned between chloroform (50 ml) and 10% aqueous citric acid solution (30 ml), and after separation of the organic layer, the aqueous layer was further extracted with chloroform (30 ml). The combined organic layer was then dried over anhydrous sodium sulfate, and after removing the drying agent by filtration, the solvent was evaporated under reduced pressure to thereby obtain crude 7- {3- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] pyrrolidine -1-yl} -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -5-hydroxy-8-methyl-1,4-dihydro-4-oxoquinoline-3-carboxylic acid.

The crude crude 7- {3- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] pyrrolidin-1-yl} -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -5 obtained above -hydroxy-8-methyl-1,4-dihydro-4-oxoquinoline-3-carboxylic acid was dissolved in concentrated hydrochloric acid (20 ml) under ice cooling, and after stirring at room temperature for 20 minutes, the solution was poured into separatory funnel and washed with chloroform (20 ml × 5 times). Then, after washing, a saturated aqueous solution of sodium hydroxide was added to the aqueous layer under cooling on ice, thereby bringing the pH to> 11, and then the pH was adjusted to 7.5-7.8 by adding concentrated hydrochloric acid and 1 mol / L chloride hydrogen acid. The resulting aqueous layer was then extracted into chloroform (100 ml), with a mixture of chloroform: methanol = 9: 1 (100 ml × 2) and the lower layer with a mixture of chloroform: methanol: water = 7: 3: 1 (100 ml), and the combined organic layer dried over anhydrous sodium sulfate. After removing the drying agent by filtration, the solvent was evaporated under reduced pressure. The residue was then purified by preparative chromatography (separation in the lower layer of a mixture of chloroform: methanol: water, 7: 3: 1), the suspension was purified using diethyl ether, and then dried under reduced pressure to obtain 119 mg (43%) of the title compound in in the form of a yellow powder.

¹ H NMR (400 MHz, 0.1 mol / L NaOD) δ: 0.45-0.55 (4H, m), 1.05-1.20 (1H, m), 1.45-1.70 (2H, m), 1.90-2.00 (1H, m), 2.05-2.20 (1H, m), 2.16 (3H, s), 3.05-3.20 (2H , m), 3.25-3.35 (1H, m), 3.40-3.50 (1H, m), 3.90-3.95 (1H, m), 4.90-5.10 (1H, m), 6.16 (1H, s), 8.33 (1H, d, J = 3.4 Hz).

Melting point: 203-206 ° C.

Specific rotation: [α] _D ^25.1 = -274.4 °.

Elemental analysis for C ₂₁ H ₂₄ FN ₃ O ₄ · 1.5 H ₂ O:

Calculated: C 58.87%; H 6.35%; N, 9.81%.

Found: C 59.23%; H 6.20%; N, 9.48%.

MS (m / z): 402 [(M + H) ⁺ ].

EXAMPLE 18

7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -8-cyano-1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro -4-oxoquinoline-3-carboxylic acid

A mixture of ethyl 8-cyano-7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carboxylate (250 mg, 0.785 mmol) , 3- (R) - [1- (tert-butoxycarbonylamino) cyclopropyl] pyrrolidine (267 mg, 1.18 mmol), 1,4-diazabicyclo [2.2.2] octane (132 mg, 1.18 mmol) and dimethyl sulfoxide (13 ml) was stirred for 2 hours at room temperature in a nitrogen-substituted atmosphere. After evaporation of the solvent, the residue was partitioned between chloroform (30 ml) and 10% aqueous citric acid solution (30 ml), and the organic layer was separated. The resulting organic layer was dried over anhydrous sodium sulfate, and after removing the drying agent by filtration, the solvent was evaporated under reduced pressure. The residue was dissolved in concentrated hydrochloric acid (4 ml) and glacial acetic acid (4 ml) at room temperature and then stirred for 12 hours while heating in an oil bath at 110 ° C. After evaporation of the solvent, concentrated hydrochloric acid (2 ml) and water (20 ml) were added, the solution was poured into a separatory funnel, and then washed with chloroform (50 ml). Then, after washing, a 10 mol / L aqueous solution of sodium hydroxide was added to the aqueous solution, thereby adjusting the pH to> 12. Then, after washing with chloroform (50 ml), the pH was adjusted to 8.3 by the addition of concentrated hydrochloric acid and 1 mol / L hydrochloric acid. The resulting aqueous layer was then concentrated to 5 ml under reduced pressure, extracted with a mixture of chloroform: methanol = 10: 1 (50 ml × 3), and the combined organic layer was dried over anhydrous sodium sulfate. After removing the drying agent by filtration, the solvent was evaporated under reduced pressure. The yellow solid thus obtained was recrystallized from ethanol and then dried under reduced pressure to obtain 124 mg (40%) of the title compound as a yellow powder.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 0.48-0.51 (4H, m), 1.66-1.90 (3H, m), 1.99-2.03 (1H, m), 2.03-2.09 (1H, m), 3.57-3.70 (3H, m), 3.79-3.83 (1H, m), 4.03-4.08 ( 1H, m), 5.18-5.35 (1H, m), 7.14 (1H, d, J = 9.3 Hz), 8.16 (1H, d, J = 9.3 Hz), 8.61 (1H, d, J = 3.9 Hz).

Melting point: 138-140 ° C.

Specific rotation: [α] _D ^24.5 = + 19.16 °.

Elemental analysis for C ₂₁ H ₂₁ FN ₄ O ₃ · 1.25 H ₂ O:

Calculated: C 60.21%; H 5.65%; N 13.07%.

Found: C, 60.42%; H 5.62%; N, 12.72%.

MS (m / z): 397 [(M + H) ⁺ ]

EXAMPLE 19

7- [3- (R) - (1-aminocyclobutyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -8-methoxy-1,4-dihydro -4-oxoquinoline-3-carboxylic acid

Mixture of 7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -8-methoxy-1,4-dihydro-4-oxoquinoline-3-carboxylic acid (290 mg, 0.98 mmol), 3- (R) - [1- (tert-butoxycarbonylamino) cyclobutyl] pyrrolidine (283 mg, 1.18 mmol), triethylamine (0.409 ml, 2.94 mmol) and dimethyl sulfoxide (5 ml) were stirred for 112 hours when heated in an oil bath at 80 ° C in a nitrogen-substituted atmosphere. After evaporation of the solvent, the residue was partitioned between chloroform (50 ml) and 10% aqueous citric acid solution (30 ml), and after the organic layer was separated, it was washed with saturated hydrochloric solution (30 ml). The resulting organic layer was then dried over anhydrous sodium sulfate, and after removing the drying agent by filtration, the solvent was evaporated under reduced pressure. The residue was purified by silica gel chromatography, and thus, chloroform: methanol = 20: 1 crude 7- {3- (R) - [1- (tert-butoxycarbonylamino) cyclobutyl] pyrrolidin-1-yl} -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -8-methoxy-1,4-dihydro-4-oxoquinoline-3-carboxylic acid.

The crude crude 7- {3- (R) - [1- (tert-butoxycarbonylamino) cyclobutyl] pyrrolidin-1-yl} -1- [2- (S)-fluoro-1- (R) -cyclopropyl] -8 obtained above -methoxy-1,4-dihydro-4-oxoquinoline-3-carboxylic acid was dissolved in concentrated hydrochloric acid (5 ml) under ice cooling, and after stirring for 5 minutes in an ice bath, the solution was poured into a separatory funnel and washed chloroform (10 ml × 3 times). Then, after washing, a 10 mmol / L aqueous solution of sodium hydroxide was added to the aqueous layer while cooling on ice, thereby bringing the pH to> 12, and then the pH was adjusted to 7.4 by adding concentrated hydrochloric acid and 1 mol / L hydrochloric acid. hydrogen acid. The resulting aqueous layer was then extracted with chloroform (100 ml × 3), and the combined organic layer was dried over anhydrous sodium sulfate. After removing the drying agent by filtration, the solvent was evaporated under reduced pressure. The residue was then recrystallized in ethyl acetate-hexane and dried under reduced pressure to give 99 mg (24%) of the title compound as a yellow powder.

¹ H NMR (400 MHz, d _6- DMSO) δ: 1.30-1.45 (1H, m), 1.45-1.60 (1H, m), 1.60-1.70 (1H, m), 1.70-2.50 (8H, m), 3.30-3.40 (1H, m), 3.40-3.50 (1H, m), 3.50 (3H, s) , 3.56-3.60 (2H, m), 4.03-4.09 (1H, m), 5.00-5.22 (1H, m), 7.09 (1H, d, J = 9.1 Hz), 7.92 (1H, d, J = 9.1 Hz), 8.57 (1H, d, J = 3.4 Hz).

Melting point: 174 ° C.

Elemental analysis for C ₂₂ H ₂₆ FN ₃ O ₄ · 0.25 H ₂ O:

Calculated: C 62.92%; H 6.36%; N, 10.01%.

Found: C 63.20%; H 6.22%; N, 10.10%.

MS (m / z): 416 [(M + H) ⁺ ].

Reference Example 50

3-cyano-2,4-difluorobenzoic acid

After dissolving diisopropylamine (56.0 ml, 395 mmol) in anhydrous tetrahydrofuran (400 ml), the solution was stirred at -15 ° C under nitrogen. After adding a hexane solution of n-butyllithium (1.52 M, 260 ml, 395 mmol) to it, the solution was stirred under cooling on ice for 1 hour. After cooling the solution to −78 ° C., a solution was added dropwise in which 2,6-difluorobenzonitrile (25.0 g, 180 mmol) was dissolved in anhydrous tetrahydrofuran (100 ml) over 1 hour. After complete addition, the reaction solution was stirred at −78 ° C. for 1 hour, and then dried carbon dioxide was bubbled into the reaction solution for 30 minutes. The reaction solution was stirred at −78 ° C. for 1 hour, then the temperature was gradually raised and then stirred at room temperature for 12 hours. Then, 100 ml of 1 mol / L hydrochloric acid was added to the reaction solution while cooling on ice and extracted with diethyl ether (500 ml × 2). The combined organic layer was washed with saturated hydrochloric solution (500 ml) and then dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, thereby obtaining 29.7 g (90%) of the yellow, amorphous title compound. This product was used in the subsequent reaction without further purification.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 7.10 (1H, m), 8.32 (1H, m).

Reference Example 51

Ethyl 8-cyano-7-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] -l, 4-dihydro-4-oxoquinoline 3-carboxylate

3-Cyano-2,4-difluorobenzoic acid (29.6 g, 162 mmol) was dissolved in dried toluene (250 ml), and after adding a catalytic amount of N, N-dimethylformamide, thionyl chloride (17.7 ml, 243 mmol) was added dropwise. ) with stirring at room temperature. After complete addition, the reaction solution was stirred for 1 hour in an oil bath at 80 ° C. Then, after the reaction solution was allowed to cool, the reaction solution was concentrated under reduced pressure, toluene (100 ml) was added to the residue, and vacuum concentration was again carried out. This operation was performed 3 times. The concentrate thus obtained was dissolved in anhydrous tetrahydrofuran (200 ml), and the solution was added dropwise with stirring and cooling on ice in a solution in which triethylamine (30 ml) and ethyl 3-dimethylaminoacrylate (24.3 g, 170 mmol) were dissolved in anhydrous tetrahydrofuran (100 ml). After complete addition, the reaction solution was refluxed for 12 hours. Then, after filtering the reaction solution through celite (washing with diethyl ether), the filtrate was concentrated under reduced pressure, and the obtained residue was purified by chromatography on a short silica gel column. Then, a brown, oily substance was obtained from the chloroform: methanol = 100: 1-100: 3 eluate by vacuum concentration.

This substance was then dissolved in anhydrous tetrahydrofuran (300 ml), 2- (S) -fluoro-1- (R) -cyclopropylamine paratoluenesulfonate (28.2 g, 114 mmol) was added and with stirring at -15 ° C, gradually added dropwise with drops a solution in which triethylamine (23 ml, 165 mmol) is dissolved in anhydrous tetrahydrofuran (50 ml). After complete addition, the reaction solution was stirred while cooling on ice for 2 hours, and then stirred at room temperature for 12 hours. Then, water (300 ml) was added to the reaction solution, and vacuum concentration was carried out to evaporate tetrahydrofuran. In addition, water (300 ml) was added and then extracted with ethyl acetate (400 ml × 3). After washing the combined organic layer with a saturated hydrochloric solution (500 ml), it was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. Thus, a tan oily substance was obtained.

This material was dissolved in dried 1,4-dioxane (400 ml) and 60% sodium hydride in oil (4.35 g) was gradually added while stirring and cooling on ice. The reaction suspension was then stirred at room temperature for 1 hour. Then, after concentrating the reaction solution to approximately 1/3 of the original volume under reduced pressure, 0.5 mol / L hydrochloric acid (50 ml) was gradually poured under cooling on ice. The precipitated solids were filtered off, washed with water and then washed with a small amount of cold ethanol and diethyl ether, in that order. The resulting crude crystals were purified by recrystallization from isopropanol and dried under reduced pressure to obtain 10.6 g (49%) of the title compound as yellowish-white crystals.

Melting point: 172-177 ° C (decomposition).

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.23-1.30 (1H, m), 1.41 (3H, t, J = 7.1 Hz), 1.61-1.99 (1H , m), 4.00 (1H, m), 4.40 (2H, q, J = 7.1 Hz), 5.10 (1H, dm, J = 63.5 Hz), 7.31 (1H , m), 8.52 (1H, d, J = 2.6 Hz), 8.77 (1H, m).

[Experimental example 1]

The antibacterial activity of the compounds of this invention was determined in accordance with standard methods developed by the Japan Society of Chemotherapy, and the results are shown as MIC values (microgram / ml) in the table below. For comparison with the MIC values of the compounds of this invention, the table also shows the MIC values of levofloxacin (LVFX), ciprofloxacin (CPFX) and 7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -6-fluoro-1 - [2- (S)-fluoro-1- (R) -cyclopropyl] -8-methyl-1,4-dihydro-4-oxoquinoline-3-carboxylic acid (reference preparation 1), which is described in PCT / JP96 / 00208.

Table 1 Example 1 Example 2 Example 3 Example 10 Example 11 E.coli, NIHJ ≦ 0.003 ≦ 0.003 ≦ 0.003 ≦ 0.003  0.006 S. flexneri, 2A 5503  0.006  0.012 ≦ 0.003  0.006  0.012 Pr. vulgalis, 08601 ≦ 0.003  0.012 ≦ 0.003  0.006  0.012 K. pneumoniae Type 1  0,025  0.05  0.012  0,025  0.1 Ser. marcescens, 10100  0,025  0.1  0,025  0.05  0.2 Ps. aeruginosa, 32104  0.05  0.2  0.05  0.1  0.2 Ps. aeruginosa, 32121  0,025  0.05  0,025  0,025  0.1 Ps. maltophilia, IID-1275  0.2  0.2  0.05  0.1  0.2 S. aureus, 209P ≦ 0.003 ≦ 0.003 ≦ 0.003 ≦ 0.003 ≦ 0.003 S. epidermidis, 56500 ≦ 0.003  0.012  0.003  0.006  0.012 Str. pyogenes, G-36 ≦ 0.003  0.006  0.003 ≦ 0.003  0.012 Str. faecalis, ATCC 19433  0.012  0,025  0.012  0.012  0.05 S. aureus, 870307  0.05  0.05  0,025  0,025  0.05 Str. pneumoniae, J24 ≦ 0.003  0.006 ≦ 0.003 ≦ 0.003  0.006 Example 14 Example 15 Reference preparation 1 LVFX CPEX E.coli, NIHJ ≦ 0.003 ≦ 0.003 ≦ 0.003  0.012 ≦ 0.003 S. flexneri, 2A 5503  0.006  0.006 ≦ 0.003  0,025  0.006 Pr. vulgalis, 08601  0.012  0.012 ≦ 0.003  0.012 ≦ 0.003 K. pneumoniae Type 1  0.05  0,025  0.006  0.01  0,025 Ser. marcescens, 10100  0.1  0.1  0.012  0.1  0,025 Ps. aeruginosa, 32104  0.1  0.1  0,025  0.2  0.005 Ps. aeruginosa, 32121  0.05  0.05  0.012  0.1  0,025 Ps. maltophilia, IID-1275  0.2  0.2 ≤0.003  0.39  0.78 S. aureus, 209P ≦ 0.003 ≦ 0.003 ≦ 0.003  0.2  0.1 S. epidermidis, 56500  0.006  0.006 ≦ 0.003  0.39  0.2 Str. pyogenes, G-36 ≦ 0.003 ≦ 0.003 ≦ 0.003  0.2  1,56 Str. faecalis, ATCC 19433  0,025  0,025  0.006  0.78  0.78 S. aureus, 870307  0.012  0,025  0.012 > 6.25  3.13 Str. pneumoniae, J24 ≦ 0.003 ≦ 0.003 ≦ 0.003  0.78  0.1

[Experimental example 2]

For the compound described in example 1 of the present invention, micronuclear analysis in the bone marrow of mice was performed as follows.

Groups of mice were used, each of which consisted of five male Slc: ddY mice six weeks old. The compound of the present invention described in example 1 and 7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -6-fluoro-1- [2- (S)-fluoro-1- (R ) -cyclopropyl] -8-methyl-1,4-dihydro-4-oxoquinoline-3-carboxylic acid (reference preparation 1) described in PCT / JP96 / 00208 was dissolved in 0.1 mol / L NaOH / saline. A solvent of 0.1 mol / L NaOH / saline was used as a control, and a drug solution obtained by dissolving and diluting cyclophosphamide (CP) in saline was used as a positive reference preparation. All drug solutions were disinfected by filtration through 0.22 μm Mylex GS filters. Each solution of the drug was administered in a single intravenous dose of 10 ml / kg at a rate of administration of 0.2 ml / min. 24 hours after administration, myeloma cells were taken from the femur, smears were prepared and stained with acrylic orange. Using a fluorescence microscope, 1000 polychromatic erythrocytes were detected in each mouse, and the frequency of micronuclear polychromatic erythrocytes and the ratio of orthochromatic and polychromatic erythrocytes per 1000 erythrocytes were calculated.

As a result, between the control and any group that was injected with 25, 50 and 100 mg / kg of the compound described in example 1, there was no significant difference in the degree of induction of micronuclei, and the evaluation result was thus negative. That is, it was found that the compound described in example 1 slightly induces the appearance of micronuclei and is safe.

Conversely, for the comparative compound 7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -6-fluoro-1- [2- (S)-fluoro-1- (R) -cyclopropyl] - 8-methyl-1,4-dihydro-4-oxoquinoline-3-carboxylic acid (reference preparation 1) described in PCT / JP96 / 00208, micronuclear induction compared to the control was clearly expressed in groups administered 50 and 100 mg / kg

These results show that the compound described in example 1 of the present invention, where the fluorine atom at position 6 of the comparative compound 7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -6-fluoro-1- [ 2- (S) -fluoro-1- (R) -cyclopropyl] -8-methyl-1,4-dihydro-4-oxoquinoline-3-carboxylic acid described in PCT / JP96 / 00208, replaced by a hydrogen atom, has a strong antibacterial activity against a wide range of both gram-negative bacteria and gram-positive bacteria, including bacteria that are resistant to antibiotics, and, in addition, is highly safe oh.

[Experimental example 3]

For the compounds of this invention described in Example 1, the concentration in the blood and the concentration in the organs after oral administration were determined using the methods described below. The same measurement methods were also performed for reference preparation 1.

Method 1: Animal Research

An injectable solution was prepared by dissolving the test compound to a concentration of 2 mg / ml (in free form) in distilled water, and using a 2.5 ml disposable syringe or metal oral probe, the solution was orally administered at a dose of 20 mg / kg to rats (Crj: CD IGS; males; 7 weeks old; Charles River Japan, Inc.) on an empty stomach.

Rats from the groups tested for absorption (4 rats per group; a total of 6 groups) were killed by bleeding under ether anesthesia 0.25, 0.5, 1, 2, 4 or 6 hours after administration, and blood, liver, kidneys and lungs were taken for samples. After coagulation, the blood was centrifuged and a serum sample was taken (3000 rpm × 15 minutes, 4 ° C). After adding 3-5 ml of 0.1 M phosphate buffer solution (pH 7.0), the tissues were homogenized and supernatants were collected after centrifugation (3000 rpm × 15 minutes, 4 ° C).

Rats from the groups studied for excretion (4 rats per group), after administration of the drug, were placed in a metabolic cage and samples were taken while cooling on ice from urine samples that were collected for 0-4 hours and 4-24 hours, and at the time of collection samples, the inside of the cell was washed with approximately 15 ml of 0.1 mol / L phosphate buffer solution (pH 7.0) in order to flush urine from the inside of the cell. Also, in order to evaluate glucuronide and other conjugated compounds, part of the sample was taken, hydrolyzed with an equivalent amount of 1 mol / L aqueous solution of sodium hydroxide, then neutralized with 0.5 mol / L hydrochloric acid, and the concentration of samples obtained by this was measured. way.

Method 2: Measure the concentration of the drug

The concentration of the drug in liquid samples was determined using the bioanalytical method of agar wells using the B. subtilis strain ATCC6051 as an organism under study. The test medium was prepared by seeding a suspension containing 5 × 10 ⁷ CFU / ml spores of the test bacteria in a proportion of 1% in food agar (Eiken Kagaku), which was sterilized at 121 ° C for 15 minutes and then cooled to about 50 ° C. After 10 ml of each given medium was poured into a sterile Petri dish and the medium solidified horizontally, four holes with a diameter of 8 mm were made in it and a system of holes was obtained for examination. To obtain a system of wells for research, the Bioassay System TDA-1 (Dainippon Seiki) was used. For measurement, test samples were prepared (diluted with serum or a solution of phosphate buffer, if necessary), serial dilutions of the drug solutions for calibration (double serial dilutions were obtained so that the diameter of the inhibitor ring was approximately 10-30 mm) and the reference drug solution (the drug solution of this concentrations to correct hole errors; typically, the concentration forming an inhibitor ring with a diameter of approximately 20 mm) and 50 μl of the test sample (or pre arat for calibration) were placed in two of four wells from each plate and 50 l of the reference solution was placed in the other two wells. After adding samples, the cup medium was allowed to stand for 1 hour at 4 ° C to effect a preliminary dispersion, then cultivation was carried out at 37 ° C for approximately 18 hours, and the diameters of the inhibitor rings were measured using CA-400 (Dainippon Seiki). The concentrations of the test samples were measured using a calibration curve obtained using second-order regression from the logarithmic values of the concentration of the drug calibration curve serial dilutions and the diameters of the inhibitor rings.

To determine tissue concentration (μg / ml), the concentration (μg / ml) of the homogenized supernatant was determined from the tissue mass (g) and the amount of added phosphate buffer (ml) using the following equation:

[Tissue concentration] = [homogenate concentration] × ([tissue weight] + [amount of buffer solution]) / [tissue weight]

Excretion in rat urine (%) was determined from the amount (μg) of the administered drug, the amount (ml) of urine (or washing solution) and the concentration (μg / ml) in the urine (washing solution):

[Excretion with rat urine] = 100 × ([amount of urine] × [concentration in urine]) / [amount of drug administered]

Method 3: Calculation of pharmacokinetic parameters

For each drug, pharmacokinetic parameters in rats were calculated based on the average concentration using a non-compartmental analysis and using the pharmacodynamics analysis program, PSAG-CP (Asmedica).

The serum concentration and concentration in organs, in the case of the liver, kidneys and lungs, of the compound of Example 1 and the reference preparation 1, which were determined by the above methods, are shown in Table 2.

table 2 Compound (addition product) The compound of example 1
(1 HCl,
0.25 IPA, 0.25 H ₂ O Reference drug
(1 MsOH,
1 H ₂ O) Serum C _max (μg / ml) 1.7 1,6 t _1/2 (h) 1,2 1,0 AUC _0-4h (mcg · h / ml) 3,1 2,3 the cloth Liver 27.4 11.6 C _max (μg / g) Kidney 14.2 5,0 Lungs 3.8 3.0 Liver 34.5 (11.0) 10.4 (4.6) AUC _0-4h (tissue / serum ratio) Kidney
Lungs 29.0 (3.2)
10.0 (3.2) 7.1 (3.1)
5.3 (3.2) Urinary excretion rate (per dose) 0-24 h
after
add conjugated
compounds 8.1
8.7 1.8
2,3

As can be seen from table 2, it was found that the compound of the present invention is distributed at higher concentrations, both in serum and in tissues, compared with reference preparation 1. Thus, it is clear that the compound of the present invention has excellent oral absorption. It is also seen that the compound of this invention penetrates excellently into tissues.

Below are the structures of the compounds whose activity was compared.

The compound described in
example 1 The compound described in PCT / JP96 / 00208
Reference preparation 1

Industrial applicability

The compounds of this invention have excellent antibacterial activity against a wide range of both gram-negative and gram-positive bacteria and, in particular, have strong antibacterial activity even against gram-positive antibiotic-resistant bacteria, such as methicillin-resistant Staphylococcus aureus (MRSA), penicillin-resistant Streptonia PRSP), vancomycin-resistant Enterococcus (VRE) and so on, and quinolone-resistant bacteria, and also has excellent safety characteristics, for example, negative in micronucleus tests, and have excellent pharmacokinetic properties such as excellent excreted in urine in rats and excellently absorbed upon oral administration and penetrates into the fabric and so forth. The compounds of the present invention are thus useful as antibacterial compounds for use in chemotherapy against microbial infections.

Claims (35)

1. The compound represented by the following General formula (I), in the form of its individual isomers or mixtures thereof, its salts and their hydrates:

where R ¹ represents a cyclic alkyl group containing 3-6 carbon atoms, which may contain a substituent selected from halogen; R ² represents a hydrogen atom; R ³ represents a hydrogen atom; R ⁴ represents a hydrogen atom, an amino group, a hydroxyl group; A represents a nitrogen atom or a part of the structure represented by formula (II)

where X ¹ represents an alkyl group containing 1-6 carbon atoms, an alkoxy group containing 1-6 carbon atoms, a hydrogen atom, an amino group, a halogen atom, a cyano group or a halogenomethoxy group, where X ¹ and the above R ¹ can be combined to form a ring structure by including part of the original skeleton, where the ring thus obtained can contain, as the ring forming atom, an oxygen atom and this ring can be replaced by an alkyl group containing 1-6 carbon atoms); each R ⁵ and R ⁶ independently represents an alkyl group containing 1-6 carbon atoms or a hydrogen atom; and n is an integer of 1 or 2. 2. The compound according to claim 1, its salts or their hydrates, where the compound of formula (I) is a stereochemically pure compound. 3. The compound according to claim 1 or 2, its salts or their hydrates, where n in the formula (I) is 1. 4. The compound according to any one of claims 1 to 3, its salts or their hydrates, where R ⁴ in the formula (I) represents a hydrogen atom. 5. The compound according to any one of claims 1 to 4, its salts or their hydrates, where A in the formula (I) represents a part of the structure represented by the formula (II). 6. The compound according to claim 5, its salts or their hydrates, where X ¹ in the formula (II) represents a methoxy group, methyl group, difluoromethoxy group, fluorine atom or chlorine atom. 7. The compound according to claim 5, its salts or their hydrates, where X ¹ in the formula (II) represents a methoxy group or a methyl group. 8. The compound according to any one of claims 1 to 7, its salts or their hydrates, where each R ⁵ and R ⁶ in the formula (I) represents a hydrogen atom. 9. The compound according to any one of claims 1 to 7, its salts or their hydrates, where one of R ⁵ or R ⁶ in the formula (I) represents a hydrogen atom and the other represents a methyl group. 10. The compound according to any one of claims 1 to 7, its salts or their hydrates, where one of R ⁵ or R ⁶ in the formula (I) represents a hydrogen atom and the other represents an ethyl group. 11. The compound according to any one of claims 1 to 10, its salts or their hydrates, where a cyclic alkyl group containing 3-6 carbon atoms, which may contain a Deputy, in R ¹ represents a halogenocyclopropyl group. 12. The compound according to claim 11, its salts or their hydrates, where the halogenocyclopropyl group is a 1,2-cis-2-halogenocyclopropyl group. 13. The compound according to item 12, its salts or their hydrates, where the halogenocyclopropyl group is a stereochemically pure substitute. 14. The compound according to item 13, its salts or their hydrates, where the halogenocyclopropyl group is a (1R, 2S) -2-halogenocyclopropyl group. 15. The compound according to any one of paragraphs.11-14, its salts or their hydrates, where the halogen atom of the halogenocyclopropyl group is a fluorine atom. 16. The compound according to claim 1, which represents 7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylic acid, its salts or their hydrates. 17. The compound according to claim 1, which represents 7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates. 18. The compound according to claim 1, which represents 7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -8-chloro-1,4-dihydro-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates. 19. The compound according to claim 1, representing 7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1-cyclopropyl-8-difluoromethoxy-1,4-dihydro-4-oxoquinoline-3 -carboxylic acid, its salts or their hydrates. 20. The compound according to claim 1, which represents 7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methoxy-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates. 21. The compound according to claim 1, which represents 7- [3- (R) - [1- (methylamino) cyclopropyl] pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R) β-cyclopropyl] -1,4-dihydro-8-methoxy-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates. 22. The compound according to claim 1, which represents 7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates. 23. The compound according to claim 1, which represents 7- [3- (R) - [1- (methylamino) cyclopropyl] pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates. 24. The compound according to claim 1, which represents 7- [3- (R) - [1- (ethylamino) cyclopropyl] pyrrolidin-1-yl] -1- [2- (S) -fluoro-1- (R) -cyclopropyl] -1,4-dihydro-8-methyl-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates. 25. The compound according to claim 1, which is 5-amino-7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R ) -cyclopropyl] -1,4-dihydro-8-methoxy-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates. 26. The compound according to claim 1, which is 5-amino-7- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -1- [2- (S)-fluoro-1- (R ) -cyclopropyl] -1,4-dihydro-8-methyl-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates. 27. The compound according to claim 1, which is 5-amino-7- [3- (R) - [1- (methylamino) cyclopropyl] pyrrolidin-1-yl] -1- [2- (S)-fluoro-1 - (R) -cyclopropyl] -1,4-dihydro-8-methyl-4-oxoquinoline-3-carboxylic acid, its salts or their hydrates. 28. The compound according to claim 1, representing 10- [3- (R) - (1-aminocyclopropyl) pyrrolidin-1-yl] -2,3-dihydro-3- (S) -methyl-7-oxo-7H -pyrido [1,2,3-de] [1,4] benzoxazine-6-carboxylic acid, its salts or their hydrates. 29. The compound according to claim 1, which is 1- (cyclopropyl) -8-methyl-7- [3- (R) - [1- (methylamino) cyclopropyl] pyrrolidin-1-yl] -1,4-dihydro- 4-oxoquinoline-3-carboxylic acid, its salts or their hydrates. 30. An antibacterial agent, which as an active component contains a compound described in any one of claims 1 to 29, its salts or their hydrates. 31. A therapeutic agent for the treatment of an infectious disease, which as an active component contains a compound described in any one of claims 1 to 29, its salts or their hydrates. 32. A method of obtaining an antibacterial agent, comprising the step of mixing the compound described in any one of claims 1 to 29, its salts or their hydrates as an active ingredient with pharmaceutically acceptable additives. 33. A method for producing a medicament for the treatment of an infectious disease, comprising the step of mixing a compound described in any one of claims 1 to 29, its salt or its hydrates as an active ingredient with pharmaceutically acceptable additives. 34. The use of the compounds described in any one of claims 1 to 29, its salts or their hydrates to obtain an antibacterial agent. 35. The use of the compounds described in any one of claims 1 to 29, its salts or their hydrates for the manufacture of a medicament for the treatment of an infectious disease.