JP5079612B2 - Antitumor agent - Google Patents

Description

  The present invention relates to an antitumor agent useful as a medicine.

  It is known that pyridonecarboxylic acid derivatives having a 2-thiazolyl group have antibacterial activity. For example, the following compound is described in Example 24 of JP-A No. 61-152682 (hereinafter referred to as Ref. 1).

  Further, Example 5 of Ref. 1 describes the following compounds.

Further, Example 12 of Ref. 1 describes the following compounds.

  The compounds represented by [Chemical Formula 2] and [Chemical Formula 3] are also described in Table 1 of JP-A-62-33176 (hereinafter referred to as Ref. 2). Example 24-4 of JP-A-60-56959 (hereinafter referred to as Ref. 3: Patent Document 3) and Example of JP-A 61-251667 (hereinafter referred to as Ref. 4: Patent Document 4) 15 describes the following compounds.

  The following compounds are described in Example 28-16 of JP-A-60-163866 (hereinafter referred to as Ref. 5: Patent Document 5).

  Furthermore, Example 8 of JP-A-2-85255 (hereinafter referred to as Ref. 6: Patent Document 6) describes the following compounds.

  However, the above Refs. 1 to 6 only mention that these compounds have an antibacterial action, and do not teach any antitumor activity or anticancer activity. These compounds are common in that a fluorine atom is substituted at the 6-position.

It is also known that certain pyridonecarboxylic acid derivatives have antitumor activity and anticancer activity. For example, Cancer Research 52 , 2818 (1992): Non-Patent Document 1 shows that as a result of examining the antitumor activity of 90 types of pyridonecarboxylic acid derivatives, most of the pyridonecarboxylic acid derivatives were found to have no antitumor activity. Reported only a few compounds. In this paper, the cyclopropyl group substituted at the 1-position and the two halogen atoms substituted at the 6-position and the 8-position play an important role in the development of antitumor action. It is taught that pyridonecarboxylic acid derivatives without substituents do not exhibit antitumor activity.

JP 61-152682 A JP-A-62-33176 JP-A-60-56959 JP 61-251667 A JP 60-163866 A JP-A-2-85255 Cancer Research 52, 2818 (1992)

  The present inventors diligently searched for a compound having an antitumor activity, and as a result, found that pyridonecarboxylic acid having no cyclopropyl group at the 1-position and related compounds have a remarkable antitumor activity, and the present invention. Was completed.

  According to the present invention, a compound having the following general formula (I) containing a 2-thiazolyl group, a stereoisomer or an optical isomer thereof, or a physiologically acceptable salt thereof, or any of the above hydrates or An antitumor agent comprising a solvate as an active ingredient is provided.

The numbers indicating the positions of the basic skeleton in the general formula (I) are, in principle, sequentially assigned counterclockwise from the position where the 2-thiazolyl group is bonded. Thus, the 2-position of thiazolyl group is attached is 1-position, a position which R ₆ is attached is a 7-position (this numbering is sometimes different from the case of usual, the specification Then, for convenience of explanation, it may be handled in this way).

  The symbols (substituents) in the general formula (I) are defined and described below.

--------- in general formula (I) means the presence or absence of a single bond. That is, the bond between the 2-position carbon atom and the 3-position carbon atom and the bond between the 4-position carbon atom and the substituent R ₄ may be a single bond or a double bond.

  X represents N or C—Rx, where Rx represents a hydrogen atom or a halogen atom. In the present specification, the term “halogen” or “halogen atom” means fluorine, chlorine, bromine or iodine, with fluorine or chlorine being preferred.

R ₁ and R ₂ are the same or different and are substituted with a hydrogen atom, a halogen atom, a nitro group, a lower alkoxy group, a lower alkyl group (the lower alkyl group may be substituted with lower alkoxycarbonyl or carboxyl), or halogen. Means a phenyl or thienyl group, which may be substituted, or a phenylsulfonyl group (wherein the phenyl moiety may be substituted with halogen or nitro), or R ₁ and R ₂ together form a benzene ring or naphthalene A ring is formed, and the benzene ring or naphthalene ring may be substituted with halogen, nitro or lower alkyl.

In this specification, unless stated otherwise, the term “lower” means that the group to which the term is attached contains 1 to 5 carbon atoms. Accordingly, for example, the lower alkyl group included in the definition of R ₁ is linear or branched, and includes, for example, methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, tert − Examples thereof include a butyl group and a pentyl group, and a methyl group and an ethyl group are preferable. The “lower alkoxy group” is a group in which the lower alkyl group and an oxygen atom are bonded, and examples thereof include a methoxy group and an ethoxy group.

R ₃ means a hydrogen atom, a benzyl group, a hydroxyl group, a carboxyl group or a group that can be converted to a carboxyl group.

The “group that can be converted into a carboxyl group” included in the definition of the substituent R ₃ may be any group that can be eliminated by chemical means or enzymatic means and converted into a carboxyl group, and is not necessarily in vivo. Need not be converted to a carboxyl group.

  As such a group that can be converted to a carboxyl group, a hydroxymethyl group, a formyl group, an ester, a carbamoyl group, or an amide is preferable.

  Examples of the ester form include lower alkyl esters such as methyl ester and ethyl ester, lower alkanoyloxy lower alkyl esters such as acetoxymethyl ester, 1-acetoxyethyl ester and pivaloyloxymethyl ester, and 1-ethoxycarbonyloxyethyl ester. Lower alkoxycarbonyloxy lower alkyl ester, di-lower alkylamino lower alkyl ester such as 2-dimethylaminoethyl ester, 2- (1-piperidinyl) ethyl ester, 3-butyrolactonyl ester, choline ester, phthalidyl ester , (5-methyl-2-oxo-1,3-dioxol-4-yl) methyl ester, and the like.

R ₄ means an oxo group or a hydroxyl group. When R ₄ is an oxo group and R ₃ is a carboxyl group, the compound represented by the general formula (I) is a pyridonecarboxylic acid derivative.

R _{5 as} a substituent at the 5-position means a hydrogen atom, an amino group, a halogen atom or a lower alkyl group, and the lower alkyl group may be substituted with a halogen. A trifluoromethyl group is mentioned as an example of the lower alkyl group substituted by the halogen here.

R ₆ means a substituent bonded to the mother nucleus by an N—C bond, a C—C bond, an S—C bond or an O—C bond, and more specifically, the following (a) to (e Examples thereof include any group selected from:

(A): R ₆ is a 4- to 7-membered monocyclic amino group represented by the following formula (a).

(Wherein --------- means the presence or absence of a single bond, R6a1, Ra2, R6a3 and R6a4 are identical or _{different, C, CH, CH 2,} CH 2 CH 2, CH ═CH, S (O) _0-2 , O, N or N—R6a, wherein R6a represents a hydrogen atom, a lower alkyl group or a phenyl group, and the phenyl group represents a lower alkoxy, lower alkyl, halogen, Or k and m may be the same or different and each represents an integer of 0 or 1. R6a5, R6a6 and R6a7 will be described later.

  The formula (a) includes, for example, a group represented by the following [Chemical 9], and these groups include an N—C bond (when m is 0) or a C—C bond (m is 1). In some cases, it is bonded to the 7th position of the mother nucleus.

  (In the formula, R6a, R6a5, R6a6, R6a7 and m are the same as described above.)

  The substituent R6a5 in the formula (a) means a hydrogen atom, a methylene group or a group represented by the following formula (R6a-5):

  [Wherein R6a51 represents a hydrogen atom or a phenyl group (the phenyl group may be substituted with halogen), or a lower alkyl group optionally substituted with phenyl or halogenophenyl; Is a hydrogen atom, a lower alkyl group or a group that can be converted to a hydrogen atom, and n is an integer of 0-4. ]

  The “group that can be converted into a hydrogen atom” included in the definition of the substituent R6a52 may be any group that can be converted into a hydrogen atom by chemical means such as hydrolysis or enzymatic means. There is no need to convert it to an atom.

  Such “groups that can be converted to hydrogen atoms” include acyl groups such as formyl and acetyl, groups having an oxycarbonyl group such as ethoxycarbonyl, amino acid residues and peptide residues, as well as, for example, o-nitrophenylsulfuric acid. Examples thereof include phenyl, trimethylsilyl, tetrahydropyranyl, diphenylphosphinyl and the like, and amino acid residues and peptide residues are particularly preferable.

  Amino acids are generally expressed in English letters consisting of three letters, and are followed in this specification. Further, the L-form, D-form or a mixture thereof is distinguished by attaching the symbols “L-”, “D-” or “DL-” to the beginning of the three letters. When these isomers are collectively referred to without distinction, these symbols are not attached.

  Examples of amino acid residues include Gly (glycine), Ala (alanine), Arg (arginine), Asn (asparagine), Asp (aspartic acid), Cys (cysteine), Glu (glutamic acid), His (histidine), Ile (Isoleucine), Leu (leucine), Lys (lysine), Met (methionine), Phe (phenylalanine), Pro (proline), Ser (serine), Thr (threonine), Trp (tryptophan), Tyr (tyrosine), Val (Valine), Nva (norvaline), Hse (homoserine), 4-Hyp (4-hydroxyproline), 5-Hyl (5-hydroxylysine), Orn (ornithine) and the like.

  Peptide residues are peptide residues in which 2 to 5, preferably 2 to 3, of the aforementioned amino acids are linked. For example, Ala-Ala, Gly-Phe, Nva-Nva, Ala-Phe, Gly- Examples include residues of peptides such as Gly, Gly-Gly-Gly, Ala-Met, Met-Met, Leu-Met, and Ala-Leu.

  The stereochemical arrangement of these amino acid and peptide residues may be D-form, L-form or a mixture thereof, but L-form is preferred. Furthermore, when the “group that can be converted to a hydrogen atom” is an amino acid or peptide residue, these residues may also have asymmetric carbon atoms. Examples of amino acid residues having an asymmetric carbon atom include amino acid residues such as Ala, Leu, Phe, Trp, Nva, Val, Met, Ser, Lys, Thr, Tyr. As an example of the peptide residue to have, what has these amino acid residues which have an asymmetric carbon atom as a structural component is mentioned.

  The substituent R6a6 included in the formula (a) is a hydrogen atom, a halogen atom, a hydroxyl group, an azide group, a formyl group, an oxo group, a lower alkoxy group, a lower alkylthio group or a lower alkyl group (the lower alkyl group is halogen or hydroxy). Which may be substituted).

  The substituent R6a7 contained in the formula (a) means the same as R6a6 or a spirocyclic amino group represented by the following formula (R6a-7):

[Wherein p represents an integer of 3 or 4. ]
R6a6 and R6a7 may be the same or different.

(B): The substituent R _{6 at the} 7-position is a bicyclic amino group represented by the following formula (b).

(Where --------- means the presence or absence of a single bond, q, s and t are the same or different and are integers from 0 to 2, and the sum of q and t is 2) or 3 .R6b1 means C, CH, a CH ₂ or N-R6b, wherein the R6b is a hydrogen atom or a lower alkyl group, R6b2 is C, CH, CH _2, CH ₂ CH ₂ or CH ═CH, R 6b3 means C, CH, CH ₂ or O, u, v and w are the same or different and are integers of 0 or 1, and their sum is 1 to 3 R6b4 and R6b5 are the same or different and each represents a hydrogen atom, a hydroxyl group, a lower alkyl group, an amino group, a mono- or di-lower alkylamino group, or a mono- or di-lower alkylamino lower alkyl group.

  In the formula (b), for example, a group represented by the following [Chemical Formula 13] is included, and these groups are bonded to the 7-position of the mother nucleus by an N—C bond.

  (Wherein ------, R6b, R6b4 and R6b5 are the same as described above.)

(C): R ₆ is a 3- to 7-membered cyclic group represented by the following formula (c).

(Wherein --------- means the presence or absence of a single bond, R6c1, R6c2, R6c3 and R6c4 are identical or _{different, C, CH, CH 2,} CH 2 CH 2, CH = CH, S (O) _0-2 , O, N or N-R6c means R6c means a hydrogen atom or a lower alkyl group. R6c5 is O, refers to S or CH _2, it is R6c6 and R6c7 same or different, represents a hydrogen atom, an amino group or a lower alkyl group. f, g and h are the same or different and mean an integer of 0 or 1. )

The formula (c) includes, for example, a group represented by the following [Chemical 15], and these groups include a C—C bond (when R6c5 is CH ₂ or h is 0), O— It is bonded to the 7-position of the mother nucleus by a C bond (when R6c5 is O and h is 1) or an S—C bond (when R6c5 is S and h is 1).

  (In the formula, R6c, R6c5, R6c6, R6c7 and h are the same as described above.)

(D): R ₆ is an amino group represented by the following formula (d).

  (Wherein R6d1 represents S or NH, R6d2 and Rd3 are the same or different, each represents a hydrogen atom or a lower alkyl group, i represents an integer of 0 or 1, and j represents an integer of 1 to 5) means.)

In formula (d), an amino group optionally substituted with lower alkyl (when i is 0), an aminoalkylamino optionally substituted with lower alkyl (when i is 1 and R6d1 is NH) and Aminoalkylthio (when i is 1 and R6d1 is S) optionally substituted with lower alkyl is included.

(E): R ₆ is any one of a hydroxyl group, a lower alkyl group, a lower alkenyl group, and a lower alkynyl group. Examples of the lower alkenyl group include a vinyl group, and examples of the lower alkynyl group include an ethynyl group.

  The compound according to the present invention represented by the general formula (I) includes a stereoisomer or an optical isomer thereof, a physiologically acceptable salt thereof, or a hydrate which may be present in some cases. Solvates are included.

  Examples of the salt include a salt of a carboxyl group part that may be present in the structure and an acid addition salt of a basic substituent part such as an amino group or a cyclic amino group.

  Examples of the salt at the carboxyl group include salts with metals such as sodium, potassium, magnesium, zinc, silver, aluminum and platinum, or organic bases such as dimethylaminoethanol, methylaminoethanol, triethanolamine and guanidine. Of the salt.

  Examples of acid addition salts with basic substituents include salts with inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, or methanesulfonic acid, oxalic acid, maleic acid. , Fumaric acid, malonic acid, lactic acid, malic acid, citric acid, tartaric acid, benzoic acid, p-toluenesulfonic acid, ascorbic acid, glucuronic acid, 2-hydroxyethanesulfonic acid, lactobionic acid, glucoheptonic acid, gluconic acid, etc. Examples include salts with acids.

  The compounds according to the present invention described above in detail are novel except for a very small part of the compounds in which the 6-position is a fluorine atom described at the beginning.

  The compounds according to the present invention are all excellent as antitumor agents or anticancer agents, and among them, compounds included in the following general formula (Ia), stereoisomers or optical isomers thereof, or physiological Or a hydrate or solvate of any of the foregoing is preferred as an antitumor agent.

(Wherein, -----, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , X, R 6a ₁ , R 6a ₂ , R 6a ₃ , R 6a ₄ , R 6a ₅ , R 6a ₆ , R 6a 7, and k are the same as described above. To do.)

  Further, the compound according to the present invention suitable as an antitumor agent is a compound represented by the following general formula (Ib), a stereoisomer or an optical isomer thereof, or a physiologically acceptable salt thereof, Any hydrate or solvate.

(Wherein, W is a hydrogen atom or a fluorine atom, and -------------------- R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R 6a ₅ , R 6a 6 and R 6a 7 are the same as described above. )

  The compound according to the present invention particularly suitable as an antitumor agent is a compound represented by the following general formula (Ic), a stereoisomer or an optical isomer thereof, or a physiologically acceptable salt thereof, or the aforementioned Any hydrate or solvate.

  (Wherein R6a511 is a lower alkyl group, and R6a61 and R6a71 are the same or different and each represents a hydrogen atom, a lower alkoxy group or a lower alkyl group.)

  The structural features of the compound represented by the general formula (Ic) are (1) based on a 1,8-naphthyridine skeleton, (2) a 2-thiazolyl group bonded to the 1-position, (3 3) a carboxyl group at position 3, (4) an oxo group at position 4, (5) no substituent such as a fluorine atom at positions 5 and 6, and (6) a specific substituent at position 7 1-pyrrolidinyl group having a hydrogen atom is bonded, in particular, a combination of substituents at the 1-position and the 7-position and a fluorine atom not contained at the 6-position.

  Specific examples of the compound included in the general formula (Ic) include 1,4-dihydro-7- (3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2 -Thiazolyl) -1,8-naphthyridine-3-carboxylic acid, its stereoisomers or optical isomers or physiologically acceptable salts thereof.

  The compounds according to the present invention are specifically described in the examples below, but in addition, the following compounds, stereoisomers or optical isomers thereof, or physiologically acceptable salts thereof, Any hydrate or solvate may be mentioned.

  7- (3-Amino-4-fluoro-1-pyrrolidinyl) -1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid

  7- (3-Amino-4-methoxy-3-methyl-1-pyrrolidinyl) -1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid

  5,8-Dihydro-2- (3-methoxy-4-methylamino-1-pyrrolidinyl) -5-oxo-8- (2-thiazolyl) pyrido [2,3-d] pyrimidine-6-carboxylic acid

  8- (4-Fluoro-2-thiazolyl) -5,8-dihydro-2- (3-methoxy-4-methylamino-1-pyrrolidinyl) -5-oxopyrido [2,3-d] pyrimidine-6-carvone acid

  5-Amino-1- (4-fluoro-2-thiazolyl) -1,4-dihydro-7- (3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1,8-naphthyridine-3 -Carboxylic acid

  5-Amino-7- (1-amino-3-azabicyclo [3.1.0] hex-3-yl) -1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8- Naphthyridine-3-carboxylic acid

  The steric structures that may be shown below are relative rather than absolute unless otherwise noted. The “stereo” column in the following table shows the relative three-dimensional structure at the 7-position, and the optically active compound is represented by (+) (−) or (R) (S).

  The compound according to the present invention is not limited to non-solid tumors such as lymphocytic leukemia tumors, but also to various solid tumors occurring in tissues such as lung, milk, stomach, uterus, skin, intestine, bladder, and nasal throat. It also shows a marked antitumor effect. In addition, the compound according to the present invention has high safety for living bodies. Therefore, the compound according to the present invention is useful as a therapeutic or prophylactic agent for human tumors.

Symbols that may be used in the following have the following meanings.
Me: methyl group,
Et: ethyl group,
Ac: an acetyl group,
Bz: benzyl group,
Ph: phenyl group,
Bu: butyl group,
t-: tert-.
[Test example]

  Next, the antitumor activity of the compounds according to the present invention will be described with reference to test examples. The commercially available anticancer agents etoposide (EP) and the commercially available antimicrobial agents ciprofloxacin (CPFX) and enoxacin (ENX) were used as controls.

Test Example 1 : Antitumor activity against P388 lymphoid mouse leukemia cells

  The antitumor activity of the test compound was tested by the MTT [3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide] method using P388 lymphoid mouse leukemia cells.

Add 0.1 ml of a culture solution containing 1000 to 2000 P388 lymphoid mouse leukemia cells and a predetermined concentration of test compound to each well of a 96-well plate, and incubate at 37 ° C. under 5% carbon dioxide for 72 hours. did. After culturing, 0.02 ml of MTT (5 mg / ml) solution was added to each well and further cultured for 4 hours. The culture solution was centrifuged (4 ° C., 2000 rpm × 20 minutes), and the supernatant was removed by aspiration. Next, 0.1 ml of dimethyl sulfoxide was added to each hole to dissolve the formed formazan, and 0.1 ml of dimethyl sulfoxide was further added. The absorbance (OD) of this solution was measured with a multi-scan bichromatic (main wavelength 570 nm, subwavelength 690 nm). The absorbance of untreated cells (control) was 100%, and a 50% growth inhibitory concentration (IC ₅₀ : μg / ml) was calculated by the least square method.

  The results are shown in Tables 1 to 9 below. The control compound data are shown in Table 9.

TABLE 1 Antitumor activity against P388 lymphoid mouse leukemia cells

*: 1 = HCl

*: 1 = HCl, 2 = 1/2 HCl, 3 = 1/5 HCl

*: 1 = HCl

*: “−” = Educt, 1 = HCl

  TABLE 2 Antitumor activity against P388 lymphoid mouse leukemia cells

*: 1 = HCl

*: “−” = Educt, 1 = HCl

*: “−” = Educt, 1 = HCl

*: “−” = Educt, 1 = HCl

*: “−” = Educt, 1 = HCl, 4 = 1/2 H ₂ SO ₄

  TABLE 3 Antitumor activity against P388 lymphoid mouse leukemia cells

*: “−” = Educt, 1 = HCl

  TABLE 4 Antitumor activity against P388 lymphoid mouse leukemia cells

*: “−” = Educt, 1 = HCl, 4 = 1/2 H ₂ SO ₄

  TABLE 5 Antitumor activity against P388 lymphoid mouse leukemia cells

  TABLE 6 Antitumor activity against P388 lymphoid mouse leukemia cells

  Continuation of Table 6

  Table 7. Antitumor activity against P388 lymphoid mouse leukemia cells

*: “−” = Educt, 1 = HCl, 5 = 4/3 CF ₃ COOH, 6 = 3/2 HCl

  TABLE 8 Antitumor activity against P388 lymphoid mouse leukemia cells

*: 1 = HCl, 7 = CF ₃ COOH

  TABLE 9 Antitumor activity against P388 lymphoid mouse leukemia cells

As shown in Tables 1 to 9, the in vitro antitumor activity (IC ₅₀ ) of the compounds according to the present invention against P388 lymphoid mouse leukemia cells is excellent. On the other hand, as shown in Table 9, ciprofloxacin and enoxacin, which are commercially available antibacterial agents, have substantially no antitumor activity.

Test Example 2 : In vitro antitumor activity against human cancer cells

0.1 ml of a culture solution containing 500 to 2000 human cancer cells was added to each well of a 96-well plate and cultured at 37 ° C. under 5% carbon dioxide for about 20 hours. After culturing, a test compound solution having a predetermined concentration was added, and further cultured for 72 hours. After the culture, 0.01 ml each of MTT (5 mg / ml) solution was added to each well, and further cultured for 4 hours. The supernatant of the culture broth was removed by suction, then 0.1 ml of dimethyl sulfoxide was added to each well to dissolve the formazan formed, and further 0.1 ml of dimethyl sulfoxide was added. For this solution, the 50% growth inhibitory concentration (IC ₅₀ : μg / ml) was calculated in the same manner as in Test Example 1, and the results shown in the following Table 10 were obtained.

The numbers in the table indicate IC ₅₀ (μg / ml).

The numbers in the table indicate IC ₅₀ (μg / ml).

As shown in Table 10, the antitumor activity (IC ₅₀ ) in vitro of the compounds according to the present invention against human cancer cells is excellent. In particular, the H-based compound according to the present invention, that is, the compound having an unsubstituted 6-position, showed an antitumor activity equivalent to or higher than that of etoposide (EP), which is a commercially available anticancer agent.

Test Example 3 : Life-prolonging effect in mice transplanted with P388 lymphoid mouse leukemia cells

1 × 10 ⁶ P388 lymphoid mouse leukemia cells were transplanted intraperitoneally into SLC: BDF ₁ mice (8-10 weeks old, female, 7 per group). The test compound (drug) was dissolved in 0.1 N-NaOH or suspended in 0.4% carboxymethylcellulose solution, which was diluted with distilled water or 0.4% carboxymethylcellulose solution to each dosage concentration. 0.2 ml of each solution was administered intraperitoneally (ip) twice the day after transplantation (day 1) and twice on day 5. The survival of mice was observed over 30 days, the median number of days of survival for each group was determined, and the survival rate (ILS:%) was calculated by the following formula.

  The drug efficacy was determined according to the standards of the National Cancer Institute in the United States. That is, the survival rate was determined to be ≧ 75%, ++ (effective); 20 to 74%, + (effective); ≦ 19%, − (ineffective). The results are shown in Table 11.

  As shown in Table 11, the life-prolonging effect of the compound according to the present invention on mice transplanted with P388 lymphoid mouse leukemia cells is excellent.

Test Example 4 : Tumor growth inhibitory effect in colon 26 mouse tumor cell transplanted mice

SLC: CDL ₁ female mice (7-9 weeks old, 7 mice per group) were transplanted with 0.1 ml of colon 26 mouse tumor cells 2% brei (brei) in the abdominal skin. The test compound (drug) was dissolved in 0.1 N NaOH and diluted with distilled water so as to have each dose concentration. 0.2 ml of each solution was administered intraperitoneally (ip) once a day from the day after transplantation (day 1) to day 9. The estimated tumor weight was calculated from the tumor diameters on the 21st to 22nd days after transplantation, and the tumor growth inhibition rate (IR:%) of the drug administration group relative to the control group was calculated from the following formula, and the results shown in Table 12 were obtained .

  As shown in Table 12, the tumor growth inhibitory effect (IR) of the compound according to the present invention on mice transplanted with colon 26 mouse tumor cells is excellent.

  In the following Test Examples 5 to 9, human cancer cells are transplanted into nude mice, and a test compound dissolved in an aqueous solution containing NaOH is administered thereto, and the strength of the degree of suppression of cancer cell proliferation is observed.

  The results of the following Test Examples 5 to 9 are shown in FIG. 1 to FIG. 5, in which the daily change of the tumor growth inhibition rate (IR) described in Test Example 4 is shown. Each figure also shows the dose of each compound (mg / kg / day) and the tumor growth inhibition rate on the last observation day.

  Here, for convenience of explanation, the administration schedule of test compounds to nude mice will be described. The test compound was administered for y days [intraperitoneal administration (ip)] x days after the day of transplantation of human cancer cells into nude mice, and then the administration was discontinued (withdrawal) for z days, and then the test compound was added again to y When administered daily, the cycle of dosing (y days) and withdrawal (z days) was termed “course”. The medication schedule in this case shall be represented by the following symbols.

  [(X) (y) (z) (cool) (ip)]

  Next, this symbol will be described with an example. However, the arrow indicates the dosing date.

  [Example] Dosing once (ip) on the 25th day after transplantation, and then once again after a 6-day drug holiday. Repeat this 5 times.

Test Example 5 : Antitumor effect on nude mice transplanted with human nasopharyngeal carcinoma cells

The experiment was conducted according to the following conditions.
Experimental conditions:
Animals used: Female BALB / c nude mice (9 to 14 weeks old, 5 to 7 animals per group).
Cancer cells used: Human nasopharyngeal carcinoma KB.
Cancer cell transplantation: 2 to 2.5 × 10 ⁶ cancer cells were transplanted into the abdominal skin of mice.
Dosing schedule: [(5) (1) (6) (6 cool) (ip)].
Result: FIG.

Test Example 6 : Antitumor effect on nude mice transplanted with human breast cancer cells

The experiment was conducted according to the following conditions.
Experimental conditions:
Animals used: Female BALB / c nude mice (7-9 weeks of age, 5-7 mice per group).
Cancer cells used: Human breast cancer MX-1.
Cancer cell transplantation: A 2 mm ³ cancer tissue fragment was implanted subcutaneously in the back of a mouse.
Medication schedule: [(15-23) (1) (6) (6 cool) (ip)].
Result: FIG.

Test Example 7 : Antitumor effect on nude mice transplanted with human colon cancer cells

The experiment was conducted according to the following conditions.
Experimental conditions:
Animals used: Female BALB / c nude mice (9 weeks old, 6 mice per group).
Cancer cells used: Human colon cancer WiDr.
Cancer cell transplantation: 2.5 × 10 ⁶ cancer cells were transplanted into the abdominal skin of mice.
Dosing schedule: [(7) (1) (6) (6 cool) (ip)].
Result: FIG.

Test Example 8 : Antitumor effect on nude mice transplanted with human melanoma

The experiment was conducted according to the following conditions.
Experimental conditions:
Animals used: Female BALB / c nude mice (8-15 weeks old, 6-7 mice per group).
Cancer cells used: Melanoma HMV-2.
Cancer cell transplantation: 2.5 to 4.4 × 10 ⁶ cancer cells were transplanted into the abdominal skin of mice.
Dosing schedule: [(8-10) (1) (6) (9 cool) (ip)].
Result: FIG.

Test Example 9 Antitumor effect on human lung cancer transplanted nude mice

The experiment was conducted according to the following conditions.
Experimental conditions:
Animals used: Female BALB / c nude mice (13 weeks of age, 6 mice per group).
Cancer cells used: Human lung cancer LX-1.
Cancer cell transplantation: A 2 mm ³ cancer tissue fragment was implanted subcutaneously in the back of a mouse.
Dosing schedule: [(19-26) (1) (6) (5-6 cool) (ip)].
Result: FIG.

  As shown in FIGS. 1 to 5 above, the compound according to the present invention remarkably suppressed the growth of human cancer cells transplanted into nude mice.

Test Example 10 : Acute toxicity

A test solution of a predetermined concentration was administered to the mouse (0.1 ml / body weight 10 g), and the LD ₅₀ value was calculated from the mortality of the mouse on the 14th day after the administration, and the results shown in the following table were obtained. In the following table, H-type compounds are toxic to female BALB / c CrSlc mice (5 to 10 mice, 10 weeks of age), and F-type compounds are female JCL: ICR mice (1 group to 5-10). ).

  As shown in Table 13, the acute toxicity of the compounds according to the present invention is comparable to that of etoposide (EP) which is a commercially available anticancer agent.

  As shown in the above test results, the compound according to the present invention is not limited to non-solid tumors such as lymphocytic leukemia tumors, but also to each tissue such as lung, milk, stomach, uterus, skin, intestine, bladder, and nasopharynx. It exhibits remarkable antitumor activity against various solid tumors that develop. Moreover, the safety | security of the compound concerning this invention is also high. Therefore, the compound according to the present invention is useful as a therapeutic or prophylactic agent for human tumors.

  The dose of the compound according to the present invention is the tumor suppressive action amount of the compound, and varies depending on the pharmacodynamic characteristics, administration method, symptom / age, treatment mode (prevention or treatment) and the like. Usually, it is about 0.4 mg to about 20 mg, preferably about 1 mg to about 10 mg, and particularly preferably 3 to 6 mg per kg body weight per day. For example, a patient weighing about 50 kg is administered a total of about 20 mg to about 1 g, preferably 50 mg to 500 mg, particularly preferably 150 mg to 300 mg of active ingredient per day. The dose may be divided into 2 to 4 times a day. The route of administration may be either oral or parenteral, but parenteral administration is recommended.

  The compounds according to the invention are generally administered in the form of preparations. These preparations can be prepared by blending the compound according to the present invention and a preparation carrier. For example, a solvent is an essential component as a formulation carrier in a liquid preparation that is a parenteral preparation, and other auxiliary components such as an isotonic agent, a solubilizer, a soothing agent, a pH adjuster, a buffer, and a preservative are included. Are appropriately blended.

  As the solvent, water is generally used, and an organic solvent such as propylene glycol or a mixture of water and an organic solvent is also used.

  Examples of the isotonic agent include saccharides such as sorbitol and mannitol, sodium chloride and the like, and saccharides are preferable.

  Examples of the pH adjuster include a base such as sodium hydroxide or an acid such as hydrochloric acid or phosphoric acid.

  Examples of the solubilizer include surfactants such as polysorbate 80 and Pluronic F68, or organic acids capable of forming an acid addition salt with a compound according to the present invention such as lactic acid and methanesulfonic acid.

  Examples of soothing agents include lidocaine hydrochloride and procaine hydrochloride; benzyl alcohol as preservatives; antioxidants such as ascorbic acid as stabilizers; acids such as phosphoric acid, citric acid, and lactic acid as buffers. And the like.

  Solutions such as injections and infusions can be prepared by dissolving or suspending, preferably dissolving, the compound according to the present invention in a solvent, and optionally adding other auxiliary components before or after dissolution or suspension. Lyophilized formulations can be prepared by lyophilizing these solutions, which are redissolved or resuspended with a solvent when administered.

  In addition, as a carrier in solid preparations such as tablets, capsules, granules, fine granules, and powders, any carrier that does not react with the compound of the present invention and is used in the art may be used. Mannit, crystalline cellulose, carboxymethylcellulose, etc. are used.

In addition, these preparations may further contain other therapeutically effective pharmaceutical ingredients, in addition to the compound according to the present invention.
[Production method]

  Next, a method for producing the compound according to the present invention will be described. The compound according to the present invention can be basically produced by a method almost similar to the method for producing antibacterial pyridonecarboxylic acid. Below, the manufacturing method by the 7-position substitution method which is a typical manufacturing method is demonstrated. Other manufacturing methods will be described in detail in the following manufacturing examples.

  The compound according to the present invention in which the 7-position is bonded to the mother nucleus through an N—C bond, an S—C bond or an O—C bond is represented by the following general formula (II)

(Wherein L is a group capable of leaving, ---, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ and X are the same as those described above) or These salts can be produced by reacting amines, thiols or alcohols corresponding to the substituent R ₆ .

  The detachable group (L) of the formula (II) may be any group that can be eliminated by substitution with the amines, such as a halogen atom, a lower alkoxy group, a lower alkylthio group, a lower alkylsulfinyl group, A lower alkylsulfonyl group, an allylsulfonyl group, a lower alkylsulfonyloxy group, an allylsulfonyloxy group and the like can be mentioned, and a halogen atom such as a fluorine atom or a chlorine atom is particularly preferable.

In addition, the amines as one raw material have a monocyclic amino group of (a) group in which m contained in the substituent R ₆ described above is 0, a bicyclic amino group of (b) group, i is Examples thereof include those corresponding to an amino group of (d) group which is 0. The thiols and alcohols, R6c5 contained in substituent R ₆ is S or O group h is is 1 (c) group and R6d1 are S i is included in a 1 (d) group The thing corresponding to is illustrated.

  The reaction of compound (II) with amines and the like can be carried out in the temperature range of 10 to 150 ° C. without solvent or in a suitable solvent, preferably in the presence of a base. As the solvent, acetonitrile, water, ethanol, pyridine, dimethyl sulfoxide, 1-methyl-2-pyrrolidone and the like are used. The base functions as an acid acceptor. For example, triethylamine, 1,8-diazabicyclo [5.4.0] -7-undecene, carbonates such as sodium carbonate and sodium hydrogen carbonate, sodium hydride, etc. are used. . When amines are used, they can be used in excess to serve as an acid acceptor.

  In addition, compound (II) used as a raw material is also novel and can be produced by, for example, the production examples described later.

  When the compound according to the present invention obtained by this method is an ester, it can be converted to a carboxylic acid by hydrolysis of the ester moiety by a conventional method. Conversely, the carboxylic acid moiety can be esterified by a conventional method or can be converted to a group that can be converted to a carboxyl group. Furthermore, when a protected amino group is present, the protecting group can be removed by a conventional method.

  The compound according to the present invention thus produced is isolated and purified according to a conventional method. Depending on the isolation and purification conditions, it can be obtained in the form of a salt, a free carboxylic acid or a free amine, but these can be converted to each other depending on the purpose to produce a desired compound.

  When the compound is a racemate, it can be separated into the respective optical isomers by applying a known method, if necessary. If necessary, the stereoisomer (cis form-trans form, R form-S form) and optically active form (+ form or -form) of the compound can be obtained by a conventional method, for example, fractional crystallization method or chromatographic method. Can be separated from each other.

  Next, an example is given and the manufacturing method of the compound concerning this invention is demonstrated in detail. In the following, the production method of intermediate (II) is used for Examples of Series A, the production method of raw material amines is used for Examples of Series B, and the target product (I) is used for Examples of Series C. The production method and the production method of the preparations in Examples of Series D will be specifically described.

Series A : Method for producing intermediate (II)

Example A-1 : --- Production of Intermediate (II)-:
7-Chloro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester

  (1) 2,6-dichloropyridine (20 g) was dissolved in tetrahydrofuran (200 ml), and an n-hexane solution (84.5 ml) of n-butyllithium (1.6 M) was added in 30 minutes at −78 ° C. in an argon stream. It was dripped. After stirring for 1 hour at the same temperature, a large excess of carbon dioxide (solid) was added. After stirring for 1 hour, the temperature was raised to −10 ° C., water and hydrochloric acid were sequentially added to adjust to pH 1-2, and the mixture was extracted with ethyl acetate. After drying over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure to dryness. Thionyl chloride (40 ml) was added to the obtained solid and heated under reflux for 3 hours. Excess thionyl chloride was distilled off under reduced pressure, and the crude product was distilled under reduced pressure to obtain 2,6-dichloronicotinic acid chloride (19.8 g).

Boiling point: 97-99 ° C./1 mmHg;
IR (neat) cm ⁻¹ : 1784

  (2) Dissolve ethoxymagnesium malonic acid diethyl ester derived from metallic magnesium (5.36 g), malonic acid diethyl ester (35.4 g) and ethanol (27 ml) in a mixed solution consisting of tetrahydrofuran (35 ml) and toluene (140 ml). To do. This solution was ice-cooled, and a mixture of tetrahydrofuran (19 ml) containing the acid chloride (44.9 g) obtained in the previous section and toluene (32 ml) was added dropwise thereto with stirring. After completion of dropping, the mixture was stirred overnight at room temperature. The mixture was concentrated under reduced pressure, aqueous hydrochloric acid was added to the residue, and the mixture was extracted with ethyl acetate. After drying this over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure to obtain an oil. Water (190 ml) and p-toluenesulfonic acid (0.1 g) were added to this and refluxed for 2 hours. After cooling, the mixture was extracted with chloroform and dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure to obtain an oil. The crude product was distilled under reduced pressure to give 2,6-dichloronicotinoyl acetic acid ethyl ester (45.2 g). Boiling point: 135-140 ° C / 2mmHg

  (3) A mixture of the compound (44.9 g) obtained in the preceding item (2), acetic anhydride (43.8 g) and ethyl orthoformate (37.7 g) was refluxed for 1 hour. The mixture was dried under reduced pressure, diisopropyl ether (500 ml) and 2-aminothiazole (20 g) were added under ice-cooling, and the mixture was stirred at room temperature for 5 hours. The precipitated crystals were collected by filtration to give 2- (2,6-dichloronicotinoyl) -3- (2-thiazolylamino) acrylic acid ethyl ester (52.8 g).

Melting point: 119-122 ° C. (recrystallized from diisopropyl ether);
IR (KBr) cm ⁻¹ : 1700

  (4) The compound (51.7 g) obtained in the preceding item (3) was dissolved in dioxane (310 ml), potassium carbonate (21.4 g) was added, and the mixture was stirred at 60 ° C. for 1 hour. After adding ice water, the mixture was neutralized with 10% aqueous hydrochloric acid, and the precipitated crystals were collected by filtration. Recrystallization from a mixture of chloroform and diisopropyl ether gave 7-chloro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester (44.6 g). Obtained.

Melting point: 176-177 ° C;
IR (KBr) cm ⁻¹ : 1724;
NMR (CDCl ₃ ) δ: 1.43 (t, 3H, J = 6.5 Hz), 4.45 (q, 2H, J = 6.5 Hz), 7.38 (d, 1H, J = 3.5 Hz), 7.52 (d, 1H, J = 8.5 Hz), 7.75 (d, 1H, J = 3.5 Hz), 8.78 (d, 1H, J = 8.5 Hz), 10.00 (s, 1H)

Example A-2 : Preparation of intermediate (II):
7-Chloro-1- (4-fluoro-2-thiazolyl) -1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylic acid ethyl ester

  A mixture consisting of the ester (250 mg) obtained in Example 1 (4), N-fluoro-2,6-dichloropyridinium tetrafluoroborate (240 mg) and 1,2-dichloroethane (10 ml) was heated to reflux for 2 days. . Water was added to the reaction mixture, and the mixture was extracted with chloroform. After drying over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: chloroform) and recrystallized from ethyl acetate to obtain the title compound (40 mg).

Melting point: 174-175 ° C;
IR (KBr) cm ⁻¹ : 1700;
NMR (CDCl ₃ ) δ: 1.42 (t, 3H, J = 6.5 Hz), 4.45 (q, 2H, J = 6.5 Hz), 7.33 (d, 1H, J = 3.5 Hz), 7.51 (d, 1H, J = 8.5 Hz), 8.78 (d, 1H, J = 8.5 Hz), 9.85 (s, 1H)

Example A-3 : Production of intermediate (II):
5,7-Dichloro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid

  (1) Sodium nitrite (3.5 g) was added to a mixture of 4-amino-2,6-dichloropyridine (5.5 g), cuprous chloride (4.4 g) and concentrated hydrochloric acid (50 ml) under ice-salt cooling. I added it little by little. After stirring at the same temperature for 1 hour and at room temperature for 1.5 hours, water was added and the mixture was extracted with chloroform. After drying over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure to obtain 2,4,6-trichloropyridine (5.5 g).

IR (neat) cm ⁻¹ : 1563, 1357, 1155, 851, 823;
NMR (CDCl ₃ ) δ: 7.31 (s, 2H)

  (2) An n-hexane solution (20 ml) of n-butyllithium (1.6 M) was added dropwise at −78 ° C. to a mixture consisting of the compound (5.5 g) obtained in the preceding item (1) and tetrahydrofuran (55 ml). After stirring for 1 hour at the same temperature, a large excess of carbon dioxide (solid) was added. After stirring for 1 hour, the temperature was raised to 0 ° C., and the mixture was acidified with aqueous hydrochloric acid and extracted with ethyl acetate. After drying over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, diisopropyl ether was added to the residue, and the crystals were collected by filtration to obtain 2,4,6-trichloronicotinic acid (6.5 g).

Melting point: 138-141 ° C;
IR (KBr) cm ⁻¹ : 1715

  (3) A mixture of the compound (6.5 g) obtained in the preceding item (2) and thionyl chloride (25 ml) was refluxed for 3 hours. Excess thionyl chloride was distilled off under reduced pressure, and the crude product was distilled under reduced pressure to obtain 2,4,6-trichloronicotinic acid chloride (6.6 g).

Boiling point: 93-95 ° C./1 mmHg;
IR (neat) cm ⁻¹ : 1791

  (4) To a mixture of malonic acid monoethyl ester (3.6 g) and tetrahydrofuran (30 ml), an ether solution (19 ml) of methylmagnesium bromide (3M) was added dropwise at 0 ° C. After stirring at room temperature for 1 hour, a mixture of the compound (6.6 g) obtained in the preceding item (3) and tetrahydrofuran (30 ml) was added dropwise and heated at 60 ° C. for 1.5 hours. The solvent was distilled off under reduced pressure, aqueous hydrochloric acid was added to the residue, and the mixture was extracted with chloroform. After drying over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the crude product was distilled under reduced pressure to obtain 2,4,6-trichloronicotinoyl acetate ethyl ester (4.8 g).

Boiling point: 160-162 ° C./2 mmHg;
IR (neat) cm ⁻¹ : 1746

  (5) A mixture consisting of the compound (4.8 g) obtained in the preceding item (4), acetic anhydride (4.2 g) and ethyl orthoformate (3.6 g) was refluxed for 1.5 hours. The mixture was dried under reduced pressure, diisopropyl ether (100 ml) and 2-aminothiazole (1.6 g) were added under ice-cooling, and the mixture was stirred at room temperature for 3 hours. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent chloroform) and recrystallized from ethyl acetate to give 2- (2,4,6-trichloronicotinoyl) -3- (2 -Thiazolylamino) acrylic acid ethyl ester (4.0 g) was obtained.

Melting point: 126-127 ° C;
IR (KBr) cm ⁻¹ : 1691

  (6) A mixture consisting of the compound (4.0 g) obtained in the previous item (5), potassium carbonate (1.5 g) and ethyl acetate (40 ml) was heated at 60 ° C. for 1 hour. The solvent was distilled off under reduced pressure, water was added to the residue, and the mixture was extracted with chloroform. After drying over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent chloroform), recrystallized from chloroform, and 5,7-dichloro-1,4-dihydro- 4-Oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester (2.5 g) was obtained.

Melting point: 226-227 ° C;
IR (KBr) cm ⁻¹ : 1737, 1692

  (7) A mixture of the ester (1.8 g) obtained in the preceding item (6) and 60 ml of 20% aqueous hydrochloric acid was heated to reflux for 5 hours. After cooling, water was added and the precipitated crystals were collected by filtration and washed with water. 5,7-Dichloro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid (1.4 g) was obtained.

Melting point: 264-266 ° C;
IR (KBr) cm ⁻¹ : 1729

Example A-4 : --Preparation of intermediate (II)-:
5-Amino-7-chloro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid

  (1) A mixture of the compound (500 mg) obtained in Example A-3 (6), benzylamine (140 mg), triethylamine (280 mg) and toluene (15 ml) was refluxed for 30 minutes. The solvent was distilled off under reduced pressure, water was added to the residue, and the mixture was extracted with chloroform. After drying over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the resulting residue was recrystallized from ethyl acetate to give 5-benzylamino-7-chloro-1,4-dihydro-4-oxo-1- (2- Thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester (510 mg) was obtained.

Melting point: 141-143 ° C;
IR (KBr) cm ⁻¹ : 1733;
NMR (CDCl ₃ ) δ: 1.42 (t, 3H, J = 7 Hz), 4.41 (q, 2H, J = 7 Hz), 4.49 (d, 2H, J = 6.5 Hz), 6.47 (s, 1H), 7.31 ( d, 1H, J = 3.5 Hz), 7.32-7.40 (m, 5H), 7.70 (d, 1H, J = 3.5 Hz), 9.87 (s, 1H), 11.2-11.7 (m, 1H)

  (2) A mixture consisting of the ester (1.0 g) obtained in the preceding item (1), concentrated sulfuric acid (2 ml) and acetic acid (8 ml) was stirred at 110 ° C. for 5 hours. After cooling, 8 ml of water was added and Stir for hours. The precipitated crystals were collected by filtration and washed with water to obtain 740 mg of the title compound.

Melting point: 264-265 ° C;
IR (KBr) cm ⁻¹ : 1727

Example A-5 : Production of intermediate (II):
7-Chloro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -5-trifluoromethyl-1,8-naphthyridine-3-carboxylic acid ethyl ester

  (1) A mixture of 2,6-dichloro-4-trifluoromethylpyridine (5 g) and tetrahydrofuran (50 ml) was cooled to -78 ° C., and n-butyllithium (1.6 M) in n-hexane (16 ml) Was added dropwise and stirred for 30 minutes. A large excess of carbon dioxide (solid) was added thereto and stirred for 1 hour. The temperature was raised to 0 ° C., extraction was performed by adding ethyl acetate and dilute hydrochloric acid, and the organic layer was dried over sodium sulfate. The solvent was distilled off under reduced pressure, thionyl chloride (20 ml) was added to the residue, and the mixture was heated to reflux for 6 hours. Excess thionyl chloride was distilled off under reduced pressure, and the residue was distilled under reduced pressure to obtain 2,6-dichloro-4-trifluoromethylnicotinic acid chloride (3.8 g).

Boiling point: 77-78 ° C./2 mmHg;
IR (neat) cm ⁻¹ : 1797

  (2) One drop of carbon tetrachloride is added to a mixture of magnesium (0.36 g) and ethanol (1.5 ml), and a mixture of diethyl malonate (2.4 g), ethanol (1.5 ml) and toluene (10 ml) is added dropwise. And stirred for 2 hours. Further, a mixture of the compound (3.8 g) obtained in the preceding item (1) and tetrahydrofuran (10 ml) was added dropwise under ice cooling, and the mixture was stirred at room temperature for 3 hours. This was extracted by adding ethyl acetate and dilute hydrochloric acid, and the organic layer was washed with water and dried over sodium sulfate, and the solvent was distilled off under reduced pressure. Water (20 ml) and p-toluenesulfonic acid (50 mg) were added to the residue, and the mixture was heated to reflux for 3 hours. Chloroform and water were added thereto for extraction, the organic layer was dried over sodium sulfate, and the solvent was distilled off under reduced pressure to obtain 2,6-dichloro-4-trifluoromethylnicotinoyl acetic acid ethyl ester (0.9 g). .

IR (neat) cm ⁻¹ : 1744, 1721;
MS (m / z): 330 (MH ^<+> )

  (3) A mixture consisting of the compound (0.9 g) obtained in the preceding item (2), ethyl orthoformate (0.6 g) and acetic anhydride (0.7 g) was heated to reflux at 140 ° C. for 1.5 hours and dried under reduced pressure. Diisopropyl ether (20 ml) was added to the residue, 2-aminothiazole (0.3 g) was added under ice cooling, and the mixture was stirred at room temperature for 3 hours. Chloroform and water were added thereto for extraction, the organic layer was dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (eluent chloroform) to give 2- (2,6-dichloro-4-trifluoromethylnicotinoyl) -3- (2-thiazolylamino) acrylic acid ethyl ester (0.37 g). It was.

IR (neat) cm ⁻¹ : 1713;
MS (m / z): 440 (MH ^<+> )

  (4) A mixture of the compound (0.37 g) obtained in the preceding item (3), potassium carbonate (0.13 g) and ethyl acetate (10 ml) was heated to reflux for 15 minutes, extracted by adding ethyl acetate and water, and the organic layer. Was dried over sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was recrystallized from ethyl acetate to give the title 7-chloro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -5-trifluoromethyl-1,8- Naphthyridine-3-carboxylic acid ethyl ester (0.27 g) was obtained.

Melting point: 184-185 ° C;
IR (KBr) cm ⁻¹ : 1736, 1703

Example A-6 : --- Production of Intermediate (II)-:
7-Chloro-6-fluoro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester

  (1) According to the method described in JP-A-61-152682 (Ref. 1), 9.8 g of 2,6-dichloro-5-fluoronicotinoyl acetate ethyl ester was treated with ethyl orthoformate and acetic anhydride. 2- (2,6-dichloro-5-fluoronicotinoyl) -3-ethoxyacrylic acid ethyl ester, and this compound was then reacted with 2-aminothiazol and triethylamine in ethanol at room temperature. To give 12.3 g of 2- (2,6-dichloro-5-fluoronicotinoyl) -3- (2-thiazolylamino) acrylic acid ethyl ester. Melting point: 125-126 ° C

(2) 12.3 g of the above compound is reacted with potassium t-butoxide in anhydrous dioxane to give 7-chloro-6-fluoro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1, 7.1 g of 8-naphthyridine-3-carboxylic acid ethyl ester was obtained.
Melting point: 182-184 ° C

Example A-7 : --- Production of Intermediate (II)-:
5,8-Dihydro-2-methanesulfonyl-5-oxo-8- (2-thiazolyl) pyrido [2,3-d] pyrimidine-6-carboxylic acid ethyl ester

  (1) A tetrahydrofuran solution (80 ml) of malonic acid monoethyl ester (12.3 g) was ice-cooled, an ether solution (64 ml) of methylmagnesium bromide (3M) was added dropwise, stirred for 20 minutes, and then 4-chloro-2 -A tetrahydrofuran solution (100 ml) of methylthiopyrimidine-5-carbonyl chloride (8.6 g) was added dropwise and stirred at room temperature for 2 hours. The reaction mixture was poured into ice water, adjusted to pH 5-6 with concentrated hydrochloric acid, and extracted with ethyl acetate. After drying over sodium sulfate, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (eluent chloroform) to give 3- (4-chloro-2-methylthiopyrimidin-5-yl) -3-oxopropionic acid ethyl ester. (8.0 g) was obtained.

IR (neat) cm ⁻¹ : 1743;
MS (m / z): 275 (MH ^<+> )

  (2) A mixture consisting of the compound (7.95 g) obtained in the preceding item (1), ethyl orthoformate (6.80 g) and acetic anhydride (7.76 g) was heated to reflux at 130 ° C. for 1 hour and concentrated under reduced pressure. Under ice-cooling, diisopropyl ether (100 ml) and 2-aminothiazole (3.28 g) were added, and the mixture was stirred overnight at room temperature. The precipitated crystals were collected by filtration and washed with diisopropyl ether. This was dissolved in 1,4-dioxane (70 ml), potassium carbonate (2.72 g) was added under ice cooling, and the mixture was stirred at room temperature for 5 hours. This was ice-cooled, ice water (200 ml) was added, and neutralized with 10% aqueous hydrochloric acid. The precipitated crystals were collected by filtration, washed successively with water, 1,4-dioxane and diisopropyl ether to give 5,8-dihydro-2-methylthio-5-oxo-8- (2-thiazolyl) pyrido [2,3- d] Pyrimidine-6-carboxylic acid ethyl ester (6.0 g) was obtained.

Melting point: 183-184 ° C;
IR (KBr) cm ⁻¹ : 1736

  (3) A methylene chloride solution (450 ml) of the compound (5.99 g) obtained in (2) above was ice-cooled, 80% m-chloroperbenzoic acid (9.30 g) was added little by little, and the mixture was stirred overnight at room temperature. did. After washing successively with aqueous sodium thiosulfate solution, aqueous sodium hydrogen carbonate solution and saturated brine, and drying over sodium sulfate, the solvent was distilled off under reduced pressure and recrystallized from a mixture of ethyl acetate and diisopropyl ether to give the title 5,8-dihydro 2-Methanesulfonyl-5-oxo-8- (2-thiazolyl) pyrido [2,3-d] pyrimidine-6-carboxylic acid ethyl ester (4.48 g) was obtained.

Melting point: 185-187 ° C;
IR (KBr) cm ⁻¹ : 1741

Example A-8 : Preparation of intermediate (II):
Intermediate (II) shown in Table 14 was obtained by the method described in Examples A-1 to A-6 or a method analogous thereto.

Table 14 Intermediate (II)

Series B : Raw material amine production method

Example B-1 : --Production of amines--:
Trans-3- (Nt-butoxycarbonylmethylamino) -4-methylpyrrolidine

  (1) Trans-3-amino-1-benzyl-4-methylpyrrolidine (19 g) was dissolved in methylene chloride (200 ml), and di-t-butyl dicarbonate (22.9 g) in methylene chloride (20 ml) was dissolved. Added under water cooling. After stirring at room temperature for 1 hour, the reaction solution was concentrated under reduced pressure to obtain trans-1-benzyl-3- (t-butoxycarbonylamino) -4-methylpyrrolidine (28.2 g).

Melting point: 138-140 ° C. (recrystallized from ethyl acetate-n-hexane);
IR (KBr) cm ⁻¹ : 3198, 1706;
MS (m / z): 291 (MH ^<+> )

  (2) A 70% toluene solution (40 ml) of sodium bis (2-methoxyethoxy) aluminum hydride was diluted with toluene (150 ml), and the compound (10 g) obtained in the preceding item (1) was gradually added under ice cooling. added. The reaction solution was heated to reflux for 1 hour, and after cooling with ice, excess reagent was decomposed with water. Insolubles were filtered off, and the filtrate was dried over anhydrous magnesium sulfate, and the solvent was evaporated under reduced pressure. The obtained residue was dissolved in methylene chloride (100 ml), and a solution of di-t-butyl dicarbonate (7.5 g) in methylene chloride (10 ml) was added to the solution under ice cooling. After stirring at room temperature for 3.5 hours, the reaction solution was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent n-hexane: ethyl acetate = 5: 1) to give trans-1-benzyl-3- (Nt-butoxycarbonylmethylamino) -4-methylpyrrolidine. (9.9 g) was obtained.

IR (neat) cm ⁻¹ : 1694;
MS (m / z): 305 (MH ^<+> )

  (3) The compound (1.52 g) obtained in the preceding item (2) was dissolved in ethanol (50 ml), 10% palladium carbon (200 mg) was added, and a theoretical amount of hydrogen was absorbed at 50 ° C. After removing the catalyst by filtration, the solvent was distilled off under reduced pressure to obtain the desired trans-3- (Nt-butoxycarbonylmethylamino) -4-methylpyrrolidine (950 mg).

IR (neat) cm ⁻¹ : 3337, 1685;
MS (m / z): 215 (MH ^<+> )

Example B-2 : Production of amines:
(+)-Trans-3- (Nt-butoxycarbonylmethylamino) -4-methoxypyrrolidine

(1) Dissolve trans-3-amino-1-benzyl-4-methoxypyrrolidine (racemate, 22.4 g) and 19.6 g of L-tartaric acid described in JP-A-2-69474 in methanol (350 ml). And left at room temperature for 7 hours. The precipitated L-tartrate was collected by filtration and recrystallized from methanol and water to obtain trans-3-amino-1-benzyl-4-methoxypyrrolidine L-tartrate (14.1 g) having the following physical properties.

  Next, all the mother liquors were combined, the solvent was distilled off under reduced pressure, saturated brine was added, the mixture was made basic with potassium carbonate, and extracted with ethyl acetate. The extract was washed with saturated brine and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The obtained residue and D-tartaric acid (6.73 g) were dissolved in methanol (180 ml) and allowed to stand at room temperature for 7 hours. The precipitated D-tartrate was collected by filtration and recrystallized from methanol and water to obtain trans-3-amino-1-benzyl-4-methoxypyrrolidine D-tartrate (9.9 g) having the following physical properties.

L-tartrate;
Melting point: 206-208 ° C (decomposition);
[Α] _D ²⁹ + 33.0 ° (c = 1.003, water);
Elemental analysis (%): C ₁₂ H ₁₈ N ₂ O 3/2 C ₄ H ₆ O ₆ ;
Calculated value C, 50.11; H, 6.31; N, 6.49;
Found C, 49.85; H, 6.26; N, 6.27;

D-tartrate;
Melting point: 207-209 ° C. (decomposition);
[Α] _D ²⁹ −33.4 ° (c = 1.020, water);
Elemental analysis (%): C ₁₂ H ₁₈ N ₂ O 3/2 C ₄ H ₆ O ₆ ;
Calculated value C, 50.11; H, 6.31; N, 6.49;
Found C, 50.35; H, 6.32; N, 6.47;

(2) Saturated saline was added to the L-tartrate (3.65 g) obtained in (1) above, neutralized with potassium carbonate, and extracted with ethyl acetate. The extract was washed with saturated brine and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to give (+)-trans-3-amino-1-benzyl-4-methoxypyrrolidine (1.23 g).
[Α] _D ²⁷ + 32.2 ° (c = 1.053, methanol)

  (3) The compound (5.74 g) obtained in (2) above was dissolved in methanol (65 ml), and di-t-butyl dicarbonate (7.29 g) was added under ice cooling. The mixture was stirred at the same temperature for 30 minutes and at room temperature for 4 hours. The solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (eluent chloroform: methanol = 50: 1) to give (+)-trans-1-benzyl-3- (t-butoxycarbonylamino) -4. -Methoxypyrrolidine (8.55 g) was obtained.

Melting point: 44-45 ° C;
[Α] _D ²⁹ +9.5 ° (c = 1.044, methanol)

(4) Lithium aluminum hydride (3.43 g) was suspended in anhydrous tetrahydrofuran (150 ml), and a solution of the compound (8.4 g) obtained in the preceding item (3) in anhydrous tetrahydrofuran (50 ml) was added dropwise, and at room temperature for 1 hour. Stir. After further refluxing for 5 hours, the excess reagent was decomposed with water under ice cooling, and the insoluble material was removed by filtration. The filtrate was extracted with ethyl acetate. The extract was washed with saturated brine and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The obtained residue was dissolved in methylene chloride (180 ml), and di-t-butyl dicarbonate (6.3 g) was added under ice cooling. After stirring at the same temperature for 30 minutes and at room temperature for 2 hours, the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent chloroform: methanol = 50: 1) to give (+)-trans-1-benzyl-3- (Nt-butoxycarbonylmethylamino) -4-methoxy. Pyrrolidine (8.26 g) was obtained.
[Α] _D ²⁹ +9.9 ° (c = 1.002, methanol)

  (5) In the same manner as in Example B-1 (3), the desired (+)-trans-3- (Nt-butoxycarbonylmethylamino) was obtained from the compound (8.15 g) obtained in (4) above. ) -4-methoxypyrrolidine (5.59 g) was obtained.

[Α] _D ²⁹ + 12.5 ° (c = 1.51, methanol);
IR (neat) cm ⁻¹ : 3318, 1693;
MS (m / z): 231 (MH ^<+> )

Example B-3 : --- Production of amines--:
(−)-Trans-3- (Nt-butoxycarbonylmethylamino) -4-methoxypyrrolidine

(1) In the same manner as in Example B-2 (2), D-tartrate (2.57 g) obtained in Example B-2 (1) was converted to (-)-trans-3-amino-1-benzyl. -4-Methoxypyrrolidine (1.01 g) was obtained.
[Α] _D ²⁷ −32.7 ° (c = 1.016, methanol)

  (2) In the same manner as in Example B-2 (3), (-)-trans-1-benzyl-3- (t-butoxycarbonylamino) -4-methoxy was obtained from the compound (3.03 g) obtained in the previous item. Pyrrolidine (4.5 g) was obtained.

Melting point: 44-45 ° C;
[Α] _D ²⁹ -9.5 ° (c = 1.080, methanol)

(3) In the same manner as in Example B-2 (4), from the compound (4.25 g) obtained in the previous item, (-)-trans-1-benzyl-3- (Nt-butoxycarbonylmethylamino)- 4-Methoxypyrrolidine (4.25 g) was obtained.
[Α] _D ²⁹ -10.1 ° (c = 1.054, methanol)

  (4) In the same manner as in Example B-2 (5), the desired (-)-trans-3- (Nt-butoxycarbonylmethylamino) -4 was obtained from the compound (4.25 g) obtained in the previous item. -Methoxypyrrolidine (2.81 g) was obtained.

[Α] _D ²⁹ -12.2 ° (c = 1.003, methanol);
IR (neat) cm ⁻¹ : 3318, 1693;
MS (m / z): 231 (MH ^<+> )

Example B-4 : --- Production of amines--:
3- (Nt-butoxycarbonylmethylamino) -3-methylpyrrolidine

  (1) In the same manner as in Example B-1 (1), 3-amino-1-benzyl-3-methylpyrrolidine (20 g) to 1-benzyl-3- (Nt-butoxycarbonylamino) -3- Methylpyrrolidine (28.3 g) was obtained.

IR (neat) cm ⁻¹ : 3356, 1716, 1697;
MS (m / z): 291 (MH ^<+> )

  (2) In the same manner as in Example B-1 (2), 1-benzyl-3- (Nt-butoxycarbonylmethylamino) -3-methylpyrrolidine was obtained from the compound (11.4 g) obtained in (1) above. (7.5 g) was obtained.

IR (neat) cm ^-1 : 1697;
MS (m / z): 305 (MH ^<+> )

  (3) In the same manner as in Example B-1 (3), the desired 3- (Nt-butoxycarbonylmethylamino) -3-methylpyrrolidine (5.5) was obtained from the compound (7.5 g) obtained in (2) above. g) was obtained.

IR (neat) cm ⁻¹ : 3337, 1682;
MS (m / z): 215 (MH ^<+> )

Example B-5 : Production of amines:
(1R ^* , 4R ^* , 5S ^* )-4-t-butoxycarbonylamino-2-azabicyclo [3.3.0] octane

  (1) To 500 g of toluene was added to 97 g of 2-ethoxycarbonylcyclopentanone, 71.2 ml of benzylamine and 5.9 g of p-toluenesulfonic acid, and heated to reflux for 2 hours while dehydrating with a Dean-Stark apparatus. The reaction solution was dried under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: n-hexane) to obtain 119.6 g of ethyl 2-benzylamino-1-cyclopentene-1-carboxylate.

Melting point: 31-32 ° C;
IR (neat) cm ⁻¹ : 3328, 2951, 1656, 1606;
MS (m / z): 246 (MH ^<+> );
NMR (CDCl ₃ ) δ: 1.25 (t, 3H, J = 7.5 Hz), 1.80 (m, 2H), 2.55 (t, 4H, J = 7.5 Hz), 4.15 (q, 2H, J = 7.5 Hz), 4.40 (d, 2H, J = 6.5Hz), 7.20-7.40 (m, 5H), 7.75 (brs, 1H)

  (2) 60 g of the compound obtained in the previous section was dissolved in 500 ml of ethanol, and 1 g of platinum dioxide was added to perform catalytic reduction at medium pressure. After removing the catalyst by filtration, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent: chloroform) to obtain 34.0 g of ethyl cis-2-benzylamino-1-cyclopentanecarboxylate. Got.

IR (neat) cm ^-1 : 2955, 2871, 1727;
MS (m / z): 248 (MH ^<+> );
NMR (CDCl ₃ ) δ: 1.25 (t, 3H, J = 7.5 Hz), 1.40-2.10 (m, 7H), 2.95 (m, 1H), 3.30 (m, 1H), 3.80 (s, 2H), 4.15 (Q, 2H, J = 7.5 Hz), 7.15-7.35 (m, 5H)

  (3) 1000 ml of dioxane was added to 85.6 g of the compound obtained in the previous section, 46 ml of ethyl bromoacetate, 57.4 g of potassium carbonate and 2.9 g of potassium iodide, and the mixture was heated to reflux for 18 hours. The reaction solution was filtered off, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent: n-hexane) to give cis-2-[(N-benzyl) ethoxycarbonylmethylamino. ] 92.5 g of ethyl 1-cyclopentanecarboxylate was obtained.

IR (neat) cm ^-1 : 2976, 2871, 1732;
MS (m / z): 334 (MH ^<+> );
NMR (CDCl ₃ ) δ: 1.10-1.30 (m, 6H), 1.50-2.10 (m, 6H), 3.10 (m, 1H), 3.35 (s, 2H), 3.50-3.70 (m, 1H), 3.85- 4.30 (m, 6H), 7.15-7.35 (m, 5H)

  (4) 12.2 g of 60% sodium hydride was suspended in 1000 ml of tetrahydrofuran, and 400 ml of a tetrahydrofuran solution containing 92.5 g of the compound obtained in the previous section was added. After heating under reflux for 1 hour, the reaction mixture was concentrated under reduced pressure, 1000 ml of 20% aqueous hydrochloric acid was added to the residue, and the mixture was heated under reflux for 6 hours. The reaction solution was neutralized and extracted with toluene. The organic layer was dried over anhydrous sodium sulfate, and then the solvent was distilled off under reduced pressure to obtain 53.7 g of cis-2-benzyl-2-azabicyclo [3.3.0] octan-4-one.

IR (neat) cm ^-1 : 2952, 2866, 1751;
MS (m / z): 216 (MH ^<+> );
NMR (CDCl ₃ ) δ: 1.50-2.05 (m, 6H), 2.75 (m, 1H), 2.80 (m, 1H), 3.15 (d, 1H, J = 16 Hz), 3.42 (d, 1H, J = 13 Hz) ), 3.45-3.55 (m, 1H), 4.03 (d, 1H, J = 13Hz), 7.20-7.40 (m, 5H)

  (5) 26 g of hydroxylamine hydrochloride was dissolved in 500 ml of water, and 200 ml of an ethanol solution of 53.7 g of the compound obtained in the previous section was added. Further, 80 ml of 20% aqueous sodium hydroxide solution was added and stirred at room temperature for 17 hours. Acetic acid water was added to the reaction solution to make it acidic, and washed with toluene. The aqueous layer was made alkaline and extracted with toluene. The organic layer was dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure to obtain 30.6 g of cis-2-benzyl-2-azabicyclo [3.3.0] octane-4-one oxime.

Melting point: 90-91 ° C. IR (KBr) cm ⁻¹ : 3279, 2958, 1699;
MS (m / z): 231 (MH ^<+> );
NMR (CDCl ₃ ) δ: 1.40-2.10 (m, 6H), 2.90-4.00 (m, 6H), 7.15-7.35 (m, 5H), 7.80 (br s, 1H)

(6) 12 g of the compound obtained in the previous item was dissolved in 120 ml of dioxane, 6 g of Raney nickel was added, and a theoretical amount of hydrogen was added at 70 ° C. After removing the catalyst by filtration, the filtrate was concentrated under reduced pressure. The resulting residue was dissolved in 100 ml of methylene chloride, and 13.6 g of di-t-butyl dicarbonate was added. After stirring at room temperature for 11 hours, the reaction mixture was concentrated under reduced pressure, ethyl acetate and diisopropyl ether were added to the resulting residue, and the precipitated crystals were collected by filtration (1R ^* , 4R ^* , 5R ^* )-2-benzyl- 10.6 g of 4-t-butoxycarbonylamino-2-azabicyclo [3.3.0] octane was obtained.

Melting point: 161-162 ° C;
IR (KBr) cm ⁻¹ : 3353, 2950, 1678;
MS (m / z): 317 (MH ^<+> );
NMR (CDCl ₃ ) δ: 1.20-1.80 (m, 6H), 1.40 (s, 9H), 2.20 (m, 1H), 3.00-3.20 (m, 2H), 3.35 (d, 1H, J = 13 Hz), 3.85 (d, 1H, J = 13Hz), 3.60-3.80 (m, 1H), 4.55 (brs, 1H), 7.15-7.35 (m, 5H)

The mother liquor obtained above was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent: chloroform) to give (1R ^* , 4S ^* , 5R ^* )-2-benzyl-4-t. -3.0 g of butoxycarbonylamino-2-azabicyclo [3.3.0] octane was obtained.

IR (neat) cm ⁻¹ : 3447, 2952, 1713;
MS (m / z): 317 (MH ^<+> );
NMR (CDCl ₃ ) δ: 1.20-1.80 (m, 6H), 1.45 (s, 9H), 2.20-2.40 (m, 1H), 2.55-3.00 (m, 3H), 3.10-3.95 (m, 2H), 4.10 (m, 1H), 4.80 (brs, 1H), 7.15-7.35 (m, 5H)

(7) Dissolve 10.5 g of (1R ^* , 4R ^* , 5R ^* )-2-benzyl-4-t-butoxycarbonylamino-2-azabicyclo [3.3.0] octane obtained in the previous section in 150 ml of ethanol. 1.0 g of 5% palladium on carbon was added and a theoretical amount of hydrogen was added. After removing the catalyst by filtration, the filtrate was concentrated under reduced pressure to obtain 7.1 g of the desired product.

Melting point: 102-103 ° C;
IR (KBr) cm ⁻¹ : 3305, 3184, 2926, 1709;
MS (m / z): 227 (MH ^<+> );
NMR (CDCl ₃ ) δ: 1.40-1.90 (m, 6H), 1.45 (s, 9H), 2.20 (m, 1H), 2.60-3.20 (m, 2H), 3.65 (brs, 1H), 3.75 (m , 1H), 4.70 (br s, 1H)

Example B-6 : --- Production of amines--:
(1R ^* , 4S ^* , 5S ^* )-4-t-butoxycarbonylamino-2-azabicyclo [3.3.0] octane

1.58 g of (1R ^* , 4S ^* , 5R ^* )-2-benzyl-4-tert-butoxycarbonylamino-2-azabicyclo [3.3.0] octane obtained in Example B-5 (6) , 200 mg of 5% palladium on carbon and 20 ml of ethanol were used in the same manner as in Example B-5 (7) to obtain 940 mg of the desired product.

Melting point: 85-87 ° C;
IR (KBr) cm ⁻¹ : 3372, 3210, 2940, 1682;
MS (m / z): 227 (MH ^<+> );
NMR (CDCl ₃ ) δ: 1.40-1.90 (m, 6H), 1.45 (s, 9H), 2.05 (brs, 1H), 2.60-3.10 (m, 3H), 3.70 (brs, 1H), 4.05 ( m, 1H), 4.75 (brs, 1H)

Example B-7 : --- Production of amines--:
1-amino-3-azabicyclo [3.1.1] heptane

  (1) 1.29 g of 60% sodium hydride was washed twice with n-hexane and then suspended in 60 ml of ether. Under cooling with ice, 5.0 g of diethyl malonate dissolved in 5 ml of ether was added dropwise over 10 minutes. This mixture was stirred at room temperature for 30 minutes and then ice-cooled. 5.38 g of benzyl chloromethyl ether dissolved in 10 ml of ether was added over 10 minutes, and the mixture was stirred overnight at room temperature and further refluxed for 5 hours. The reaction mixture was poured into ice water, extracted with ethyl acetate, washed with saturated brine, and dried over sodium sulfate. The solvent was distilled off under reduced pressure to obtain 9.0 g of a crude product of diethyl benzyloxymethylmalonate.

  This was dissolved in 50 ml of ether and added dropwise to a mixture of 2.85 g of lithium aluminum hydride and 300 ml of ether under ice cooling. The mixture was stirred at the same temperature for 1.5 hours and further at room temperature for 2 hours. Under ice-cooling, saturated aqueous sodium tartrate was added and stirred overnight. Insoluble material was filtered off, and the filtrate was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent chloroform: ethanol = 40: 1 to 20: 1) to obtain 1.96 g of 2-benzyloxymethyl-1,3-propanediol.

IR (neat) cm ⁻¹ : 3380;
¹ H-NMR (CDCl ₃ ) δ: 2.05 (m, 1H), 2.35 (brs, 2H), 3.63 (d, 2H, J = 5.5 Hz), 3.81 (dd, 4H, J = 5.5, 5.5 Hz) , 4.52 (s, 2H), 7.25-7.40 (m, 5H)

  (2) While cooling a mixture of 84.8 g of the compound obtained in the previous section and 360 ml of pyridine with ice-salt, 206.3 g of p-toluenesulfonyl chloride was added in three portions and stirred at the same temperature for 2 hours and at room temperature overnight. did. The reaction mixture was poured into ice water and extracted with ethyl acetate. After washing with dilute hydrochloric acid and water, it was dried over sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was purified by silica gel chromatography (eluent chloroform: ethanol = 30: 1) to obtain 162.3 g of 2-benzyloxymethyl-1,3-bis (p-toluenesulfonyloxy) propane. It was.

IR (neat) cm ⁻¹ : 1598, 1361;
¹ H-NMR (CDCl ₃ ) δ: 2.33 (m, 1H), 2.44 (s, 6H), 3.41 (d, 2H, J = 6 Hz), 4.04 (m, 4H), 4.34 (s, 2H), 7.10 -7.40 (m, 9H), 7.74 (d, 4H, J = 8Hz)

  (3) 22.8 g of 60% sodium hydride was washed twice with n-hexane and then suspended in 350 ml of toluene. Under cooling with ice, 91.1 g of diethyl malonate dissolved in 150 ml of toluene was added dropwise over 25 minutes. After stirring this mixture at room temperature for 40 minutes, 119.6 g of the compound obtained in the previous item dissolved in 300 ml of toluene was added and refluxed for 4.5 days. The reaction mixture was poured into ice water, extracted with ethyl acetate, washed with saturated brine, and dried over sodium sulfate. The solvent and unreacted diethyl malonate were distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: chloroform: ethanol = 50: 1) to give diethyl 3-benzyloxymethylcyclobutane-1,1-dicarboxylate. 66.8 g of ester was obtained.

IR (neat) cm ⁻¹ : 1726, 1263;
¹ H-NMR (CDCl ₃ ) δ: 1.23 (t, 3H, J = 7 Hz), 1.25 (t, 3H, J = 7 Hz), 2.25-2.50 (m, 2H), 2.55-2.80 (m, 3H), 3.44 (d, 2H, J = 6Hz), 4.17 (q, 2H, J = 7Hz), 4.21 (q, 2H, J = 7Hz), 4.50 (s, 2H), 7.20-7.40 (m, 5H)

  (4) 66.8 g of the compound obtained in the previous item was dissolved in 500 ml of ethyl acetate, 6.68 g of 5% palladium carbon was added, and a theoretical amount of hydrogen was added at 30 to 40 ° C. After removing the catalyst by filtration, the solvent was distilled off, and the residue was purified by silica gel chromatography (eluent chloroform: ethanol = 20: 1) to obtain 45.0 g of 3-hydroxymethylcyclobutane-1,1-dicarboxylic acid diethyl ester. It was.

IR (neat) cm ⁻¹ : 3452, 1728, 1266;
¹ H-NMR (CDCl ₃ ) δ: 1.25 (t, 3H, J = 7 Hz), 1.27 (t, 3H, J = 7 Hz), 1.70 (br s, 1H), 2.30-2.50 (m, 2H), 2.50 -2.70 (m, 3H), 3.62 (d, 2H, J = 5.5 Hz), 4.20 (q, 2H, J = 7 Hz), 4.23 (q, 2H, J = 7 Hz)

  To a mixture of 45.0 g of 3-hydroxymethylcyclobutane-1,1-dicarboxylic acid diethyl ester obtained in the previous section and 350 ml of pyridine, 48.5 g of p-toluenesulfonyl chloride was added in four portions under ice cooling. After stirring at the same temperature for 5.5 hours, at room temperature for 13.5 hours, and at 30 to 40 ° C. for 8 hours, ice water was added and the mixture was extracted with ethyl acetate. The extract was washed successively with saturated aqueous sodium hydrogen carbonate, water, dilute hydrochloric acid and saturated brine, and dried over sodium sulfate. The solvent was distilled off under reduced pressure to obtain a crude product of 3-p-toluenesulfonyloxymethylcyclobutane-1,1-dicarboxylic acid diethyl ester.

  A mixture of the above crude product, 42.2 g of benzylamine, 32.4 g of potassium carbonate and 600 ml of dioxane was refluxed for 4.5 days. Ice water was added to the reaction mixture, and the mixture was extracted with chloroform, washed with saturated aqueous sodium hydrogen carbonate and then saturated brine, and dried over sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent chloroform: ethanol = 20: 1) to obtain 32.3 g of 3-benzylaminomethylcyclobutane-1,1-dicarboxylic acid diethyl ester. It was.

IR (neat) cm ⁻¹ : 3324, 2813, 1727, 1263;
¹ H-NMR (CDCl ₃ ) δ: 1.23 (t, 3H, J = 7 Hz), 1.25 (t, 3H, J = 7 Hz), 2.15-2.30 (m, 2H), 2.40-2.70 (m, 3H), 2.67 (d, 2H, J = 6Hz), 3.76 (s, 2H), 4.17 (q, 2H, J = 7Hz), 4.21 (q, 2H, J = 7Hz), 7.20-7.40 (m, 5H)

  (5) A mixture comprising 32.3 g of 3-benzylaminomethylcyclobutane-1,1-dicarboxylic acid diethyl ester obtained in the preceding paragraph, 120 ml of 10% aqueous sodium hydroxide solution and 120 ml of tetrahydrofuran was stirred at room temperature for 13.5 hours and then 50-60 Stir at 6 ° C. for 6 hours. The solution was concentrated under reduced pressure, filtered with 100 ml of water and activated carbon. To the filtrate, 54.5 ml of 20% aqueous hydrochloric acid was added at room temperature with stirring. After cooling with ice, the crystals were collected by filtration and washed with water and ethanol to obtain 20 g of 3-benzylaminomethylcyclobutane-1,1-dicarboxylic acid.

Melting point: 224-226 ° C (decomposition);
IR (KBr) cm ⁻¹ : 3444, 1632;
¹ H-NMR (DMSO-d ₆ ) δ: 2.10-2.26 (m, 2H), 2.30-2.45 (m, 2H), 2.65-2.95 (m, 1H), 3.02 (d, 2H, J = 7 Hz), 4.11 (s, 2H), 7.35-7.55 (m, 5H), 9.00 (br s, 2H)

  (6) 37 g of carbonyldiimidazole was added to a mixture consisting of 20 g of the compound obtained in the previous section and 200 ml of N, N-dimethylformamide, and the mixture was stirred at room temperature for 13 hours, 5.5 hours at 30-40 ° C, and stirred at 60-70 ° C for 4 hours did. The solvent was distilled off under reduced pressure, and diluted hydrochloric acid was added to the residue to adjust to pH 2. The mixture was extracted with ethyl acetate, washed with saturated brine, and dried over sodium sulfate. The solvent was distilled off under reduced pressure, diisopropyl ether-n-hexane (2: 1) was added to the residue, the precipitated crystals were collected by filtration, and 3-benzyl-2-oxo-3-azabicyclo [3.1.1] heptane- 15.5 g of 1-carboxylic acid was obtained.

Melting point: 127-128 ° C (recrystallized from ethyl acetate);
IR (KBr) cm ⁻¹ : 2531, 1750, 1732;
¹ H-NMR (CDCl ₃ ) δ: 1.85-2.00 (m, 2H), 2.70-2.84 (m, 1H), 2.84-2.97 (m, 2H), 3.44 (d, 2H, J = 2 Hz), 4.65 ( s, 2H), 7.20-7.45 (m, 5H)

  (7) A mixture consisting of 15.5 g of the compound obtained in the previous section, 9.6 g of triethylamine and 150 ml of t-butanol was heated to 70 ° C., and 26 g of diphenylphosphoryl azide was added dropwise. The reaction was refluxed for 15 hours. The mixture was concentrated under reduced pressure, ice water was added to the residue, and the mixture was extracted with ethyl acetate. The extract was washed with dilute aqueous sodium hydroxide solution and then with saturated brine, and then dried over sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (eluent chloroform: ethanol = 50: 1) to give 3-benzyl-1-t-butoxycarbonylamino-3-azabicyclo [3.1.1]. 14.6 g of heptan-2-one was obtained.

Melting point: 106-107 ° C (recrystallized from methylene chloride-diisopropyl ether);
IR (KBr) cm ⁻¹ : 3402, 1723, 1646;
¹ H-NMR (CDCl ₃ ) δ: 1.45 (s, 9H), 1.70-1.83 (m, 2H), 2.56 (tt, 1H, J = 2.5, 7 Hz), 3.10-3.35 (m, 2H), 3.25 ( d, 2H, J = 2.5Hz), 4.60 (s, 2H), 6.22 (brs, 1H), 7.20-7.40 (m, 5H)

  (8) A mixture consisting of 6.07 g of the compound obtained in the previous item, 12 ml of 10% aqueous hydrochloric acid and 6 ml of tetrahydrofuran was heated at 50 to 60 ° C. for 3 hours. The mixture was concentrated under reduced pressure, and a small amount of water was added to the residue, followed by washing with n-hexane. The aqueous layer was made alkaline with aqueous sodium hydroxide and extracted with chloroform. The extract was washed with saturated brine and dried over sodium sulfate. The solvent was distilled off under reduced pressure to obtain 4.09 g of 1-amino-3-benzyl-3-azabicyclo [3.1.1] heptan-2-one.

IR (neat) cm ⁻¹ : 3381, 1659, 1495, 1250;
¹ H-NMR (CDCl ₃ ) δ: 1.85-2.00 (m, 2H), 1.90 (s, 2H), 2.10-2.25 (m, 2H), 2.54 (tt, 1H, J = 2.5, 7 Hz), 3.26 ( d, 2H, J = 2.5Hz), 4.60 (s, 2H), 7.20-7.40 (m, 5H)

  (9) 30 ml of a 1.0 M tetrahydrofuran solution of borane-tetrahydrofuran complex was added to a mixture of 2.1 g of the compound obtained in the previous section and 25 ml of tetrahydrofuran. After refluxing for 7 hours, the mixture was concentrated under reduced pressure. 40 ml of ethanol was added to the residue and refluxed for 9 hours. The mixture was concentrated under reduced pressure, ice water was added to the residue, and the mixture was extracted with ethyl acetate. The extract was washed with saturated aqueous sodium hydrogen carbonate and then with saturated brine, and then dried over sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was distilled with a Kugelrohr microdistiller to obtain 1.86 g of 1-amino-3-benzyl-3-azabicyclo [3.1.1] heptane.

Boiling point: about 100 ° C (3mmHg);
IR (neat) cm ⁻¹ : 3360, 2791, 1603;
¹ H-NMR (CDCl ₃ ) δ: 1.51 (br s, 2H), 1.60-1.90 (m, 4H), 2.18 (tt, 1H, J = 3, 6 Hz), 2.65 (s, 2H), 2.69 (d , 2H, J = 3Hz), 3.67 (s, 2H), 7.15-7.40 (m, 5H)

  (10) 1.86 g of the compound obtained in the previous section was dissolved in 20 ml of ethanol, 190 mg of 5% palladium carbon was added, and a theoretical amount of hydrogen was added. The catalyst was removed by filtration, and the solvent was distilled off under reduced pressure to obtain 1.0 g of the desired product.

Example B-8 : Production of amines:
9-methylene-1,7-diazaspiro [4.4] nonane dihydrochloride

(1) A mixture consisting of 300 g of 2,3-dibromopropene, 245.3 g of 3,4-dimethoxybenzylamine, 503.7 g of triethylamine and 3000 ml of toluene was heated to reflux for 45 minutes. After cooling, water and aqueous acetic acid were added, and ethyl acetate was added. And washed with water. After drying over anhydrous sodium sulfate, the solvent was distilled off and the residue was purified by silica gel column chromatography (eluent chloroform) to obtain 120 g of 2-bromo-3- (3,4-dimethoxybenzylamino) propene.
IR (neat) cm ⁻¹ : 2934, 2833, 1516

(2) N- (t-butoxycarbonyl) -L-proline 73.8 g, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (WSC) 72.4 g and 1000 ml of dichloromethane Below, a mixture of 108 g of the compound obtained in the previous section and 100 ml of dichloromethane was added dropwise over 20 minutes, and the mixture was stirred at room temperature for 20 hours. Water was added to the mixture, followed by extraction with dichloromethane. After drying over anhydrous sodium sulfate, the solvent was distilled off, and the residue was purified by silica gel column chromatography (eluent chloroform) to give N- (2-bromopropenyl) -N- (3,4-dimethoxybenzyl) -1- 149 g of (t-butoxycarbonyl) prolinamide was obtained.
IR (neat) cm ^-1 : 2974, 1697

  (3) To a mixture consisting of 67.7 g of the compound obtained in the previous section and 800 ml of toluene, 800 ml of a toluene solution of 0.5 N potassium hexamethyldisilazane was added dropwise over 20 minutes under ice cooling, and the mixture was stirred at the same temperature for 90 minutes. Water and aqueous acetic acid were added thereto, and the mixture was extracted with ethyl acetate. The extract was washed with saturated brine, dried over anhydrous sodium sulfate, the solvent was evaporated, the residue was purified by silica gel column chromatography (eluent chloroform), and 1- (t-butoxycarbonyl) -7- (3.4 29.3 g of -dimethoxybenzyl) -9-methylene-6-oxo-1,7-diazaspiro [4.4] nonane was obtained.

IR (neat) cm ⁻¹ : 2971, 1692;
MS (m / z): = 403 (MH ^<+> )

  (4) To a mixture consisting of 20 g of the compound obtained in the previous section and 500 ml of dichloromethane, 100 ml of trifluoroacetic acid was added dropwise over 10 minutes under ice cooling, followed by stirring at room temperature for 16 hours. Water and an aqueous sodium hydroxide solution were added thereto, and the mixture was extracted with dichloromethane. After drying over anhydrous sodium sulfate, the solvent was distilled off, and the residue was purified by silica gel column chromatography (eluent chloroform) to give 7- (3,4-dimethoxybenzyl) -9-methylene-6-oxo-1,7. -10.6 g of diazaspiro [4.4] nonane was obtained.

IR (neat) cm ⁻¹ : 2959, 1699;
MS (m / z): 303 (MH ^<+> )

  (5) 20.5 ml of a 70% toluene solution of bis (2-methoxyethoxy) aluminum sodium hydride was added dropwise over 5 minutes to a mixture consisting of 10.6 g of the compound obtained in the previous section and 200 ml of toluene at room temperature. Stir for 5 hours. To this were added brine and aqueous sodium hydroxide solution, and the mixture was extracted with ethyl acetate. After washing with saturated brine, the solution was dried over anhydrous sodium sulfate, and the solvent was distilled off. The residue was purified by silica gel column chromatography (eluent chloroform) to obtain 7.8 g of 7- (3,4-dimethoxybenzyl) -9-methylene-1,7-diazaspiro [4.4] nonane.

IR (neat) cm ⁻¹ : 2953, 1516;
MS (m / z): 289 (MH ^<+> )

  (6) A mixture consisting of 0.23 g of the compound obtained in the previous item, 0.57 g of 1-chloroethyl chloroformate and 10 ml of 1,2-dichloroethane was heated to reflux for 2 hours, and the solvent was distilled off. 10 ml of methanol was added to the residue, heated under reflux for 1 hour, and the solvent was distilled off to obtain 0.12 g of the desired 9-methylene-1,7-diazaspiro [4.4] nonane dihydrochloride. This compound was used as a production raw material of compound 6F-6 described later.

IR (neat) cm ⁻¹ : 3400, 2917, 1631;
MS (m / z): 139 (MH ^<+> );
¹ H-NMR (DMSO-d ₆ ) δ: 1.8-2.1 (m, 4H), 3.0-3.3 (m, 4H), 3.6.-3.9 (m, 2H) 5.26 (t, 1H, J = 2 Hz), 5.43 (t, 1H, J = 2Hz)

Example B-9 : --- Production of amines--:
3-hydroxy-3-trifluoromethylpyrrolidine

(1) To 14.5 g of 1-benzyl-3-pyrrolidone, 150 ml of tetrahydrofuran and 0.92 ml of 1M tetrahydrofuran solution of tetra-n-butylammonium fluoride were added. Under ice cooling, 16.5 g of trimethylsilyltrifluoromethane was added to the mixture and stirred for 30 minutes. The reaction solution was brought to room temperature and further stirred for 3 hours. Thereto was added 30 ml of a 1M tetrahydrofuran solution of tetra-n-butylammonium fluoride, and the mixture was stirred at room temperature for 4 hours. After completion of the reaction, water and ethyl acetate were added for extraction. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (eluent chloroform) to obtain 13.3 g of 1-benzyl-3-hydroxy-3-trifluoromethylpyrrolidine.
Melting point 60-61 ° C

  (2) To 1.0 g of the compound obtained in the previous section, 15 ml of ethanol and 100 mg of 10% palladium carbon were added, and a theoretical amount of hydrogen was absorbed at 60 ° C. The catalyst was removed by filtration, and the solvent was evaporated to dryness to obtain 630 mg of the target amine.

Example B-10 : Production of amines:
(1R, 5S, 8R) -8- (tert-butoxycarbonylamino) -2-oxa-6-azabicyclo [3.3.0] octane

(1) (3S, 4R, 5S) -1-benzyl-3,4-bis (tert-butyl) produced according to the description of J. Org. Chem. 60 , 103-108 (1995) using D-tartaric acid as a raw material. To 125.5 g of (dimethylsilyloxy) -5- (2-hydroxyethyl) -2-pyrrolidinone, 1200 ml of 60% aqueous acetic acid was added and refluxed overnight. After concentration under reduced pressure, 300 ml of concentrated aqueous ammonia and 500 ml of methanol were added to the residue, and the mixture was stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent chloroform: methanol = 10: 1) to give (3S, 4R, 5S) -1-benzyl-3,4-dihydroxy-5- ( 35 g of 2-hydroxyethyl) -2-pyrrolidinone was obtained.

IR (neat) 3370, 1682 cm ^-1 ;
MS (m / z): 252 (MH ⁺ )

¹ H-NMR (CDCl ₃ ) δ: 7.10-7.32 (m, 5H), 5.52 (br s, 2H), 4.90 (d, 1H, J = 15.0Hz), 4.50 (d, 1H, J = 7.5Hz) , 4.21 (br t, 1H, J = 7.5Hz), 3.94 (d, 1H, J = 15.0Hz), 3.55 (br s, 3H), 2.20-2.00 (br s, 1H), 1.86 (br s, 2H )

  (2) 35 g of the compound obtained in the previous section was added to 400 ml of pyridine, and 26.6 g of p-toluenesulfonyl chloride was added to the ice-cooled place and stirred overnight. Water and chloroform were added to the reaction solution, and the product was extracted into an organic layer, washed with 10% aqueous hydrochloric acid and dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure to obtain 24 g of oily (1R, 5S, 8S) -6-benzyl-8-hydroxy-7-oxo-2-oxa-6-azabicyclo [3.3.0] octane.

IR (neat): 3390, 1692 cm ^-1 ;
MS m / z: 234 (MH ⁺ )

¹ H-NMR (CDCl ₃ ) δ: 7.40-7.18 (m, 5H), 4.84 (d, 1H, J = 15.0Hz), 4.49 (br, 1H), 4.42 (dd, 1H, J = 6.5, 1.7Hz ), 4.32 (br, 1H), 4.16-4.06 (m, 2H), 3.92-3.66 (m, 2H), 1.92-1.82 (m, 2H)

(3) 14 g of the compound obtained in the previous section was added to 250 ml of methylene chloride, and 14.6 ml of pyridine was added to the ice-cooled place, and then 13.1 ml of trifluoromethanesulfonic anhydride was slowly added. Two hours later, 550 ml of dimethylformamide and 58.9 g of potassium acetate were added to the reaction solution and stirred overnight. Insoluble matter was filtered and water and chloroform were added to the residue obtained by concentration under reduced pressure. The product was extracted into an organic layer, and then the solvent was distilled off under reduced pressure to obtain crude (1R, 5S, 8S) -8-. Acetoxy-6-benzyl-7-oxo-2-oxa-6-azabicyclo [3.3.0] octane was obtained.

  To this compound, 500 ml of etal and 200 ml of concentrated aqueous ammonia were added, stirred overnight at room temperature, and concentrated under reduced pressure. Water and chloroform were added to the residue, and the product was extracted into an organic layer. The solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (eluent chloroform: methanol = 30: 1) (1R, 5S, 8R). 9.2 g of -6-benzyl-8-hydroxy-7-oxo-2-oxa-6-azabicyclo [3.3.0] octane was obtained.

IR (neat): 3381, 1694 cm ^-1 ;
MS m / z: 234 (MH ⁺ )

¹ H-NMR (CDCl ₃ ) δ: 7.40-7.20 (m, 5H), 4.93 (d, 1H, J = 15.0Hz), 4.50 (dd, 1H, J = 6.0, 5.0Hz), 4.28 (d, 1H , J = 6.0Hz), 4.09 (d, 1H, J = 15.0Hz), 4.08-3.93 (m, 2H), 3.80-3.66 (m, 1H), 3.22 (br s, 1H), 2.24-2.11 (m , 1H), 2.01-1.81 (m, 1H)

  (4) 9.2 g of the compound obtained in the previous item was added to 100 ml of methylene chloride, 9.5 ml of pyridine was added to the ice-cooled place, and then 9.1 ml of trifluoromethanesulfonic anhydride was slowly added. After 2 hours, 300 ml of dimethylformamide and 24.4 g of sodium azide were added to the reaction solution and stirred overnight. Water and chloroform were added to the residue obtained by concentration under reduced pressure, and the product was extracted into an organic layer. After washing with dilute hydrochloric acid water and drying over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and (1R, 5S, 8S) -8-azido-6-benzyl-7-oxo-2-oxa-6-azabicyclo [3 .3.0] 7.4 g of octane was obtained.

IR (neat): 2109, 1694 cm ^-1 ;
¹ H-NMR (CDCl ₃ ) δ: 7.40-7.20 (m, 5H), 4.93 (d, 1H, J = 15.0Hz), 4.23 (dd, 1H, J = 6.0, 1.5Hz), 4.16-4.02 (m , 3H), 3.94-3.82 (m, 1H), 3.81-3.67 (m, 1H), 2.00-1.80 (m, 2H)

  (5) 7.4 g of the compound obtained in the previous section was added to 300 ml of tetrahydrofuran, and 116 ml of 1.0 mol-tetrahydrofuran solution of borane-tetrahydrofuran complex was added to the solution after ice cooling. Heated after 30 minutes and refluxed overnight. The reaction was cooled to room temperature and excess borane was treated with ethanol. After concentration under reduced pressure, 600 ml of ethanol was added to the residue and refluxed overnight. After concentration under reduced pressure, 10% aqueous hydrochloric acid was added to the residue, and the product was extracted into an aqueous layer and washed with chloroform. The mixture was made alkaline with 20% aqueous NaOH and extracted with chloroform. After drying over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure to obtain 5.5 g of (1S, 5S, 8R) -8-amino-6-benzyl-2-oxa-6-azabicyclo [3.3.0] octane. .

  Subsequently, this compound was added to 100 ml of methanol and cooled on ice, and 7.2 g of dicarbonate-di-tert-butyl dicarbonate was added thereto and stirred overnight. The residue obtained by concentration under reduced pressure was subjected to silica gel column chromatography (eluent n-hexane: ethyl acetate = 6: 1) to give (1S, 5S, 8R) -6-benzyl-8- (tert-butoxycarbonylamino). ) -2-oxa-6-azabicyclo [3.3.0] octane 7.0 g was obtained.

Melting point 115-116 ° C. (recrystallization from ethyl acetate);
[Α] _D ²⁷ + 2.3 ° (c = 1.024, methanol)

  (6) 5.4 g of the compound obtained in the previous section was dissolved in 100 ml of ethanol, 350 mg of 5% palladium carbon was added, and a theoretical amount of hydrogen was absorbed at 40 ° C. After removing the catalyst by filtration and distilling off the solvent under reduced pressure, the resulting crude crystals were recrystallized from ethyl ether-diisopropyl ether to obtain the final target product (1R, 5S, 8R) -8- (tert-butoxycarbonyl). 3.5 g of amino) -2-oxa-6-azabicyclo [3.3.0] octane was obtained.

Melting point 110-111 ° C .;
IR (KBr): 3377, 3228, 1680 cm ^-1 ;
[Α] _D ²⁹ -44.4 ° (c = 1.026, methanol)

Series C : Method of manufacturing the target

Example C-1 : --- Production of object (I)-:
1,4-Dihydro-7- (trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid (racemate) ) And related compounds

(1) 7-Chloro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester obtained in Example A-1 (4) (7.1) Triethylamine (18 ml) was added to a suspension consisting of g) and trans-3-methoxy-4-methylaminopyrrolidine dihydrochloride (6.0 g) and acetonitrile (150 ml). The mixture was stirred at room temperature for 5 hours, the reaction mixture was concentrated under reduced pressure, an aqueous sodium hydrogen carbonate solution was added, and the mixture was extracted with chloroform. The extract was dried over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent chloroform: methanol = 6: 1) to give 1,4-dihydro-7- ( Trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester (6.1 g) was obtained.
Melting point: 73-76 ° C

(2) A solution consisting of the ester (6.0 g) and 18% aqueous hydrochloric acid (100 ml) was stirred at 100 ° C. for 28 hours. The precipitated crystals were collected by filtration, washed with a mixture of ethanol and diisopropyl ether, and 4.45 g of 1,4-dihydro-7- (trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo- 1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid hydrochloride (Compound H-1-1-1) (4.45 g) was obtained.
Melting point: 270-273 ° C

(3) A solution consisting of the hydrochloride (51.5 g) obtained in (2), water (500 ml) and aqueous ammonia (40 ml) was stirred at 50 ° C. overnight. Acetonitrile was added thereto, and the mixture was concentrated under reduced pressure, and the precipitated crystals were collected by filtration. The obtained crystals were washed with water and acetonitrile to obtain 32.6 g of 1,4-dihydro-7- (trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2-thiazolyl). ) -1,8-naphthyridine-3-carboxylic acid (Compound H-1-1-0) was obtained.
Melting point: 290-292 ° C. (decomposition)

(4) A mixture composed of the compound (2.0 g) obtained in the preceding item (3), lactic acid (3.1 g) and distilled water (4 ml) was heated and dissolved at 60 ° C. After cooling to room temperature, ethanol (70 ml) was added and the precipitated crystals were collected by filtration and washed with ethanol to obtain 2 g of lactate (Compound H-1-1-2).
Melting point: 288-291 ° C (decomposition)

  (5) 1.27 g of WSC was added to a mixture consisting of the ester (1.89 g), Nt-butoxycarbonyl-L-alanine (1.26 g) and methylene chloride (80 ml) obtained in the preceding item (1). Stir for hours. After washing with water, it was dried over anhydrous sodium sulfate and concentrated under reduced pressure. 7- {trans-3- [N- (Nt-butoxycarbonyl-L-alanyl)] methylamino-4-methoxy-1 was purified by silica gel column chromatography (eluent chloroform: methanol = 50: 1). -Pyrrolidinyl} -1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester (2.15 g) was obtained.

Melting point: 120-123 ° C;
[Α] _D ²⁸ + 10.2 ° (c = 1.0, chloroform)

  A mixture of this ethyl ester (1.71 g), 0.5 N aqueous hydrochloric acid (42 ml), and ethanol (22 ml) was stirred with heating at 80 ° C. for 17.5 hours. The solution was concentrated under reduced pressure, and the precipitated crystals were collected by filtration, washed with 10% aqueous hydrochloric acid and ethanol, and washed with 7- [trans-3- (N-L-alanylmethylamino) -4-methoxy-1-pyrrolidinyl]. 1.16 g of -1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid hydrochloride was obtained.

Melting point: 230-233 ° C;
[Α] _D ²⁹ +8.4 ° (c = 1.0, water)

Example C-2 : --- Production of object (I)-:
(+)-1,4-dihydro-7- (trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carvone Acid (optically active substance) and related compounds

(1) Implementation with 7-chloro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester obtained in Example A-1 (4) (−)-7 in the same manner as in Example C-1 (1) using (+)-trans-3- (Nt-butoxycarbonylmethylamino) -4-methoxypyrrolidine obtained in Example B-2. -[Trans-3- (Nt-butoxycarbonylmethylamino) -4-methoxy-1-pyrrolidinyl] -1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine- 3-carboxylic acid ethyl ester (non-crystal) was obtained.
[Α] _D ²⁹ -9.1 ° (c = 1.006, chloroform)

  (2) The title hydrochloride (Compound H-1-2) was obtained from the ethyl ester obtained in (1) above according to the method described in Example C-1 (2).

Melting point: 278-282 ° C (decomposition);
[Α] _D ²⁹ + 24.8 ° (c = 0.500, water)

  (3) A solution consisting of the hydrochloride (28.1 g) obtained in (2), water (300 ml) and aqueous ammonia (25 ml) was stirred at 50 ° C. overnight and then concentrated under reduced pressure. Precipitated crystals were collected by filtration and washed with water and ethanol to obtain the target carboxylic acid compound (Compound H-1-2-1) (19.5 g).

Melting point: 268-271 ° C;
[Α] _D ³⁰ + 53.1 ° (c = 1.005, 1N NaOH)

(4) 6.22 g of the carboxylic acid compound obtained in the above item (3) was dissolved in 150 ml of distilled water, and 14.8 ml of 1.002 N-sodium hydroxide aqueous solution was added under ice cooling, followed by immediate stirring for 10 minutes. The solvent was distilled off under reduced pressure, acetonitrile and diisopropyl ether were added to the residue, and the precipitated crystals were collected by filtration to obtain 6.28 g of the intended sodium salt (Compound H-1-2-2).
Melting point: 189-190 ° C (decomposition)

  (5) Methanesulfonic acid (140 mg) was added to a suspension composed of the compound (300 mg) obtained in (3) above and water (1.2 ml), and dissolved by heating at 60 ° C. Filtered hot and the filtrate was cooled at room temperature. Acetone (10 ml) was added thereto, and the precipitated crystals were collected by filtration and washed with acetone to obtain the methanesulfonic acid salt of the title compound (Compound H-1-2-3).

Melting point: 283-285 ° C. (decomposition);
[Α] _D ²⁸ + 46.9 ° (c = 1.00, 1N NaOH)

  (6) The lactate salt of the title compound (Compound H-1-2-4) was obtained from the compound obtained in the above (3) according to the method described in Example C-1 (4).

Melting point: 273-275 ° C. (decomposition);
[Α] _D ²⁷ + 44.7 ° (c = 1.00, 1N NaOH)

  (7) The compound (350 mg) obtained in the above (3) and gluconic acid (about 50%, 1.1 g) were dissolved in water (1 ml) by heating (60 ° C.). After allowing to cool, acetone (100 ml) is added, and the precipitated solid is recrystallized from water-methanol. The precipitated crystals are collected by filtration, washed with methanol and the gluconate of the title compound (Compound H-1-2-5). )

Melting point: 129-132 ° C;
[Α] _D ²⁸ + 36.7 ° (c = 0.36, 1N NaOH)

  (8) The compound obtained in (1) above was treated with hydrochloric acid-ethanol (30%) to give (−)-1,4-dihydro-7- (trans-3-methoxy-4-methylamino-1- Pyrrolidinyl) -4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester (Compound H-1-2-6) was obtained.

Melting point: 184-185 ° C;
[Α] _D ²⁸ −16.9 ° (c = 0.503, chloroform)

Example C-3 : --- Production of object (I)-:
(-)-1,4-dihydro-7- (trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carvone Acid (optically active substance) and its salt

(1) Implementation with 7-chloro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester obtained in Example A-1 (4) (+)-7 in the same manner as in Example C-1 (1) using (−)-trans-3- (Nt-butoxycarbonylmethylamino) -4-methoxypyrrolidine obtained in Example B-3 -[Trans-3- (Nt-butoxycarbonylmethylamino) -4-methoxy-1-pyrrolidinyl] -1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine- 3-carboxylic acid ethyl ester (non-crystal) was obtained.
[Α] _D ²⁹ +9.0 ° (c = 1.002, chloroform)

  (2) According to the method described in Example C-1 (2), the hydrochloride of the title compound (Compound H-1-3) was obtained from the ethyl ester obtained in (1) above.

Melting point: 278-282 ° C (decomposition);
[Α] _D ²⁹ -25.2 ° (c = 0.504, water)

  (3) The target carboxylic acid compound (Compound H-1-3-1) was obtained from the compound obtained in (2) above according to the method described in Example C-2 (3).

Melting point: 277-279 ° C. (decomposition);
[Α] _D ²⁹ -59.9 ° (c = 1.00, 1N NaOH)

  (4) According to the method described in Example C-2 (5), the methanesulfonate (Compound H-1-3-3) of the title compound was obtained from the compound obtained in (3) above.

Melting point: 287-289 ° C. (decomposition);
[Α] _D ³⁰ −47.2 ° (c = 1.00, 1N NaOH)

Example C-4 : --- Production of object (I)-:
1,4-dihydro-7- (cis-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid and related Compound

(1) 7-chloro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester obtained in Example A-1 (4) and cis 1,4-Dihydro-7- (cis-3-methoxy-4-methylamino-1-pyrrolidinyl)-in the same manner as in Example C-1 (1) using 3-methoxy-4-methylaminopyrrolidine 4-Oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester was obtained.
Melting point 253-259 ° C. (decomposition)

(2) According to the method described in Example C-1 (2), the hydrochloride of the title compound (Compound H-1-4) was obtained from the ethyl ester obtained in (1) above.
Melting point: 263-269 ° C. (decomposition)

Example C-5 : --- Production of object (I)-:
7- (3-Amino-1-pyrrolidinyl) -6-fluoro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid hydrochloride (Compound 2F- 1)

(1) Ethyl 7-chloro-6-fluoro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylate 6.5 obtained in Example A (6) A mixture of g, 3-aminopyrrolidine (1.7 g), triethylamine (5.6 g) and acetonitrile (65 ml) was heated to reflux for 30 minutes. After cooling, the crystals are collected by filtration and recrystallized from ethanol to give 7- (3-amino-1-pyrrolidinyl) -6-fluoro-1,4-dihydro-4-oxo-1- (2-thiazolyl). ) -1,8-naphthyridine-3-carboxylic acid ethyl ester (ester of compound 2F-1) 7.1 g was obtained.
Melting point: 218-220 ° C.

(2) A mixture of 7.1 g of the ester and 200 ml of 10% aqueous hydrochloric acid was heated to reflux for 1 hour. After cooling, the crystals are collected by filtration and recrystallized from water-ethanol to give 6.9 g of 7- (3-amino-1-pyrrolidinyl) -6-fluoro-1,4-dihydro-4-oxo- 1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid hydrochloride (Compound 2F-1) was obtained.
Melting point: 294-296 ° C (decomposition)

(3) The above hydrochloride is treated according to a conventional method, and 7- (3-amino-1-pyrrolidinyl) -6-fluoro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8 -Naphthyridine-3-carboxylic acid (compound 2F-1 free form) was obtained.
Melting point: 253-254 ° C. (decomposition)

  (4) (R) -3-Aminopyrrolidine was used and reacted and treated as described above to obtain the following compound.

7-[(R) -3-Amino-1-pyrrolidinyl] -6-fluoro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester ;
Melting point: 254-257 ° C

  7-[(R) -3-Amino-1-pyrrolidinyl] -6-fluoro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid hydrochloride (Compound 2F-1-1)

Melting point: 297-300 ° C. (decomposition);
[Α] _D ²⁸ = −21.0 ^O (1N NaOH, c = 0.1)

  (5) (S) -3-Aminopyrrolidine was used and reacted and treated in the same manner as above to obtain the following compound.

7-[(S) -3-Amino-1-pyrrolidinyl] -6-fluoro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester ;
Melting point: 256-259 ° C

  7-[(S) -3-Amino-1-pyrrolidinyl] -6-fluoro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid hydrochloride (Compound 2F-1-2)

Melting point:> 300 ° C;
[Α] _D ²⁸ = + 20.0 ^O (1N NaOH, c = 0.1)

(6) 7- [3- (N-L-alanylamino) -1-pyrrolidinyl] -6-fluoro-1,4-dihydro-4-oxo-1 according to the description in Example C-1 (5) -(2-Thiazolyl) -1,8-naphthyridine-3-carboxylic acid hydrochloride (Compound 2F-1-3) was obtained. ;
Melting point: 236-239 ° C

Example C-6 ; --- Production of object (I)-:
The compounds shown in Tables 15 to 21 below and the ethyl esters thereof were obtained by the methods described in Examples C-1 to C-5 or a method analogous thereto.

  Table 15: Target compound (I)

*: 1 = HCl, 2 = 1/2 HCl, 3 = 1/5 HCl
**: (minutes) indicates the decomposition point. The same applies hereinafter.

*: 1 = HCl

*: “−” = Educt, 1 = HCl

Table 16: Target compound (I)

*: 1 = HCl

*: “−” = Educt, 1 = HCl

*: “−” = Educt, 1 = HCl

*: “−” = Educt, 1 = HCl

*: “−” = Educt, 1 = HCl, 4 = 1/2 H ₂ SO ₄

*: “−” = Educt, 1 = HCl

  Table 17: Target compound (I)

*: “−” = Educt, 1 = HCl

  Table 18: Objective Compound (I)

*: “−” = Educt, 1 = HCl, 4 = 1/2 H ₂ SO ₄

*: “−” = Educt, 1 = HCl,
**: 3-position ethyl ester

  Table 19: Target compound (I)

  Continuation of Table 19

  Table 20: Target compound (I)

  Continuation of Table 20

  Continuation of Table 20

  Table 21. Target compound (I)

*: Measured in water (c = 0.1).
**: Measured in DMSO (c = 0.1).

*: Measured in water (c = 0.1).
**: Dihydrochloride.

*: Measured in water (c = 0.1).
**: Measured in water (c = 0.5).

Example C-7 : --- Production of object (I)-:
2- (3-Amino-1-pyrrolidinyl) -5,8-dihydro-5-oxo-8- (2-thiazolyl) pyrido [2,3-d] pyrimidine-6-carboxylic acid

5,8-Dihydro-2-methanesulfonyl-5-oxo-8- (2-thiazolyl) pyrido [2,3-d] pyrimidine-6-carboxylic acid ethyl ester and 3-amino obtained in Example A-7 Pyrrolidine was reacted in the same manner as in Example C-1 to obtain the title compound (9N-1).
Melting point: 288-291 ° C (decomposition)

Example C-8 : --- Production of object (I)-:
3-Formyl-1,4-dihydro-7- (trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2-thiazolyl) -1,8-naphthyridine hydrochloride (compound 7H-1)

  (1) 7-chloro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester obtained in Example A-1 (4) (8.49) g) To a suspension consisting of trans-3-methoxy-4-methylaminopyrrolidine dihydrochloride (6.5 g) and acetonitrile (400 ml) was added triethylamine (13.4 ml). After stirring overnight at room temperature, the reaction mixture was concentrated under reduced pressure, an aqueous sodium hydrogen carbonate solution was added, and the mixture was extracted with chloroform. The extract was washed with saturated brine and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. Methylene chloride (500 ml) was added to the resulting residue, and di-t-butyl dicarbonate (6.4 g) was added under ice cooling. After stirring at room temperature overnight, the reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent chloroform: methanol = 100: 1) to give 7- [trans-3- (Nt-butoxy). Carbonylmethylamino) -4-methoxy-1-pyrrolidinyl] -1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester (9.1 g) It was.

IR (KBr) cm ⁻¹ : 1735, 1695;
MS (m / z): 530 (MH ^<+> )

  (2) To a mixture of the compound (8.95 g) obtained in the preceding item (1) and ethanol (150 ml), 1N aqueous sodium hydroxide solution (150 ml) was added under ice cooling, and the mixture was stirred at the same temperature for 30 minutes and at room temperature overnight. did. The solution was acidified with aqueous acetic acid and extracted with chloroform. After washing with saturated brine and drying over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure to remove 7- [trans-3- (Nt-butoxycarbonylmethylamino) -4-methoxy-1-pyrrolidinyl] -1, 4-Dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid (7.19 g) was obtained.

Melting point 185-188 ° C;
IR (KBr) cm ⁻¹ : 1715, 1690;
MS (m / z): 502 (MH ^<+> )

  (3) Sodium borohydride (2.16 g) was added to a mixture consisting of the compound (7.15 g) obtained in the preceding item (2) and methanol (400 ml) under ice cooling, and the same temperature was kept for 15 minutes at room temperature overnight. Stir. The solvent was removed under reduced pressure and then extracted with chloroform. The extract was washed with saturated brine and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent chloroform: methanol = 100: 1) to give 7- [trans-3- (Nt-butoxycarbonylmethylamino) -4-methoxy-1-pyrrolidinyl]. -1,2,3,4-Tetrahydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine (3.9 g) was obtained.

IR (neat) cm ⁻¹ : 1690;
MS (m / z): 460 (MH ⁺ )

  (4) The compound (3.8 g) obtained in (3) above was dissolved in tetrahydrofuran (500 ml), and an n-hexane solution (6.1 ml) of n-butyllithium (1.6 M) was added dropwise at −78 ° C. After stirring at the same temperature for 30 minutes, ethyl formate (1.34 ml) was added, the temperature was raised slowly, and the mixture was stirred overnight. The solvent was distilled off under reduced pressure, aqueous acetic acid was added to the residue, and the mixture was extracted with chloroform. After washing with saturated brine and drying over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure to remove 7- [trans-3- (Nt-butoxycarbonylmethylamino) -4-methoxy-1-pyrrolidinyl] -3- Formyl-1,2,3,4-tetrahydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine (3.87 g) was obtained.

IR (neat) cm ⁻¹ : 1690, 1615;
MS (m / z): 488 (MH ^<+> )

  (5) The compound (3.6 g) obtained in (4) above is dissolved in 1,4-dioxane (160 ml), and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (2.51 g) Was added slowly. After stirring for 2 hours, the solvent was distilled off under reduced pressure, an aqueous sodium hydroxide solution was added to the residue, and the mixture was extracted with chloroform. After washing with saturated brine and drying over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent chloroform: methanol = 100: 1) to give 7- [trans- 3- (Nt-butoxycarbonylmethylamino) -4-methoxy-1-pyrrolidinyl] -3-formyl-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine ( 1.73 g) was obtained.

Melting point 130-132 ° C .;
IR (KBr) cm ⁻¹ : 1695, 1645, 1615;
MS (m / z): 486 (MH ^<+> )

  (6) A mixture composed of the compound (1.65 g) obtained in the preceding item (5), 10% aqueous hydrochloric acid (10 ml) and ethanol (40 ml) was heated at 50-60 ° C. for 7 hours. The precipitated crystals were collected by filtration and washed with ethanol and diisopropyl ether to give 3-formyl-1,4-dihydro-7- (trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2-Thiazolyl) -1,8-naphthyridine hydrochloride (Compound 7H-1) (1.05 g) was obtained.

Melting point 255-263 ° C. (decomposition);
IR (KBr) cm ⁻¹ : 3460, 1695, 1645;
MS (m / z): 386 (MH ^<+> )

Example C-9 : --- Production of object (I)-:
1,2,3,4-tetrahydro-4-hydroxy-3-hydroxymethyl-7- (trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -1- (2-thiazolyl) -1,8- Naphthyridine (Compound 7H-2)

  Under ice cooling, 4.90 g of 1,4-dihydro-7- (trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2-thiazolyl) -1,8- Naphthyridine-3-carboxylic acid ethyl ester was dissolved in 150 ml of anhydrous tetrahydrofuran, 3.00 g of lithium aluminum hydride was added, stirred for 30 minutes, and further stirred overnight at room temperature. The reaction solution was slowly added to water, and an aqueous ammonium chloride solution and an aqueous sodium hydrogen carbonate solution were sequentially added. This was extracted with chloroform, washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (eluent chloroform: methanol = 50: 1) to obtain the target compound 7H-2 (1.60 g). Melting point: 69-70 ° C

Example C-10 : --- Production of object (I)-:
1,4-dihydro-7- (trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid 2-hydroxy Ethyl ester 4/3 trifluoroacetate (compound 7H-3)

(1) 1.00 g of 7- [trans-3- (Nt-butoxycarbonylmethylamino) -4-methoxy-1-pyrrolidinyl] -1,4-dihydro-4-oxo-1 under ice cooling -(2-Thiazolyl) -1,8-naphthyridine-3-carboxylic acid was dissolved in 90 ml of methylene chloride, 1.24 g of ethylene glycol, 420 mg of WSC and 50 mg of 4-dimethylaminopyridine were added and stirred for 30 minutes, and further at room temperature. Stir overnight. Water was added, this was extracted with chloroform, washed with saturated brine, and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (eluent chloroform: methanol = 50: 1), and 1,4-dihydro-7- [trans-3- (Nt-butoxycarbonylmethylamino) -4-methoxy-1 -Pyrrolidinyl] -4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid 2-hydroxyethyl ester (590 mg) was obtained.
Melting point: 101-105 ° C

(2) 250 mg of the compound of the previous item was dissolved in 3 ml of methylene chloride, and 1.0 ml of trifluoroacetic acid was added under ice cooling, followed by stirring for 1 hour and further stirring at room temperature overnight. Diisopropyl ether was added and the precipitated crystals were collected by filtration to obtain the target compound 7H-3 (150 mg).
Melting point: 125-127 ° C

Example C-11 : --- Production of object (I)-:
1- [1,4-Dihydro-7- (trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carbonyl] -3-Hydroxypyrrolidine 3/2 hydrochloride (Compound 7H-4)

(1) Under ice-cooling, 250 mg of 7- [trans-3- (Nt-butoxycarbonylmethylamino) -4-methoxy-1-pyrrolidinyl] -1,4-dihydro-4-oxo-1- (2-Thiazolyl) -1,8-naphthyridine-3-carboxylic acid was dissolved in 20 ml of methylene chloride, and 52 mg of 3-pyrrolidinol and 265 mg of benzotriazole-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate (BOP reagent) were dissolved therein. Then, 0.17 ml of triethylamine was added and stirred for 30 minutes, and further stirred overnight at room temperature. Water was added, this was extracted with chloroform, washed with saturated brine, and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (eluent chloroform: methanol = 20: 1) to give 1- {7- [trans-3- (Nt-butoxycarbonylmethylamino) -4-methoxy-1-pyrrolidinyl]. -1,4-Dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carbonyl} -3-hydroxypyrrolidine (280 mg) was obtained.
Melting point: 127-129 ° C

(2) 275 mg of the compound of the previous item was dissolved in 5 ml of ethanol, 3.0 ml of 10% ethanol-hydrochloric acid was added under ice cooling, and the mixture was stirred for 1 hour, and further stirred at room temperature overnight. Diisopropyl ether was added and the precipitated crystals were collected by filtration to obtain the target compound 7H-4 (163 mg).
Melting point: 189-193 ° C (decomposition)

Example C-12 : --- Production of object (I)-:
3-hydroxymethyl-1,4-dihydro-7- (trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2-thiazolyl) -1,8-naphthyridine hydrochloride ( Compound 7H-5)

110 mg of 3-formyl-1,4-dihydro-7- (trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1 obtained in Example C-8 under ice cooling -(2-thiazolyl) -1,8-naphthyridine hydrochloride was dissolved in 30 ml of methanol, 108 mg of cerium chloride heptahydrate and 30 mg of sodium borohydride were added thereto, and the mixture was stirred for 30 minutes. Chloroform and aqueous acetic acid solution were added. added. The solution was made alkaline with an aqueous sodium hydrogen carbonate solution and extracted with chloroform. The extract was washed with saturated brine, dried over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure, an ethanol-hydrochloric acid solution was added, and the precipitated crystals were collected by filtration to obtain 69 mg of the target compound 7H-5.
Melting point: 263-268 ° C (decomposition)

Example C-13 : --- Production of object (I)-:
7- (3-Amino-1-pyrrolidinyl) -6-fluoro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxamide trifluoroacetate (Compound 8F -1)

(1) Under ice cooling, 9.00 g of 7- [3- (Nt-butoxycarbonylamino) -1-pyrrolidinyl] -6-fluoro-1,4-dihydro-4-oxo-1 in 150 ml of methanol -(2-Thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester, 1N-aqueous sodium hydroxide solution (150 ml) was added, and the mixture was stirred for 30 minutes and further stirred at room temperature overnight. Acetic acid aqueous solution was added, and the mixture was extracted with chloroform. After washing with saturated brine, the organic layer was dried over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and 7- [3- (Nt-butoxycarbonylamino) -1-pyrrolidinyl] -6-fluoro-1,4- 8.50 g of dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid was obtained.
Melting point: 220-224 ° C

(2) 2.20 g of the compound of the preceding item was dissolved in 20 ml of chloroform, 0.97 ml of triethylamine and 604 mg of ethyl chloroformate were added under ice cooling, and the mixture was stirred for 1 hour and further stirred at room temperature for 1 hour. Again, under ice cooling, 20 ml of a saturated ammonia-ethanol solution was added and stirred for 1 hour, and further stirred overnight at room temperature. The solvent was distilled off under reduced pressure, water was added to the residue, and the mixture was extracted with chloroform. It dried with the anhydrous sodium sulfate and the solvent was depressurizingly distilled. The residue was purified by silica gel column chromatography (eluent chloroform: methanol = 100: 1) and 7- [3- (Nt-butoxycarbonylamino) -1-pyrrolidinyl] -6-fluoro-1,4-dihydro 1.19 g of -4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxamide was obtained.
Melting point: 240-243 ° C

(3) 700 mg of the compound of the previous item was dissolved in 40 ml of chloroform, and 1.3 ml of trifluoroacetic acid was added under ice cooling, followed by stirring for 1 hour and further stirring at room temperature overnight. The solvent was distilled off under reduced pressure, ethanol was added to the residue, and the precipitated crystals were collected by filtration to obtain 635 mg of the target compound (8F-1).
Melting point: 227-233 ° C. (decomposition)

Example C-14 : --- Production of object (I)-:
7- (3-Amino-1-pyrrolidinyl) -6-fluoro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine trifluoroacetate (Compound 8F-2)

(1) 7- [3- (Nt-butoxycarbonylamino) -1-pyrrolidinyl] -6-fluoro-1,4-dihydro-4-oxo-1- (2-thiazolyl) under ice cooling 11.3 g of -1,8-naphthyridine-3-carboxylic acid was dissolved in 400 ml of methanol, and 4.5 g of sodium borohydride was added and stirred for 30 minutes, and further stirred overnight at room temperature. The solvent was distilled off under reduced pressure, aqueous acetic acid was added to the residue, and the mixture was extracted with chloroform. The extract was washed with saturated brine and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (eluent chloroform: methanol = 100: 1), and 7- [3- (Nt-butoxycarbonylamino) -1-pyrrolidinyl] -6-fluoro-1,2,3. , 4-Tetrahydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine 5.41 g was obtained.
Melting point: 234-236 ° C

(2) 700 mg of the compound of the previous item was dissolved in 70 ml of 1,4-dioxane, 440 mg of 2,3-dichloro-5,6-dicyano-1,4-benzoquinone was added, and the mixture was heated to reflux overnight. The solvent was distilled off under reduced pressure, an aqueous sodium hydroxide solution was added to the residue, and the mixture was extracted with chloroform. It was dried over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (eluent chloroform: methanol = 100: 1) to give 7- [3- (Nt-butoxycarbonylamino)- 1-Pyrrolidinyl] -6-fluoro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine (270 mg) was obtained.
Melting point: 234-236 ° C

(3) 160 mg of the compound of the previous item was dissolved in 12 ml of chloroform, and 0.6 ml of trifluoroacetic acid was added under ice cooling, followed by stirring for 1 hour and further stirring at room temperature overnight. The solvent was distilled off under reduced pressure, ethanol was added to the residue, and the precipitated crystals were collected by filtration to obtain 150 mg of the target compound (8F-2).
Melting point: 237-240 ° C

Example C-15 : --- Production of object (I)-:
7- (3-Amino-1-pyrrolidinyl) -3-benzyl-6-fluoro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine (Compound 8F-3)

(1) 7- [3- (Nt-Butoxycarbonylamino) -1-pyrrolidinyl] -6-fluoro-1,2,3,4-tetrahydro-obtained in Example C-14 (1) in 10 ml of ethanol 400 mg of -4-oxo-1- (2-thiazolyl) -1,8-naphthyridine and 5 ml of 20% aqueous sodium hydroxide solution were added, 350 mg of benzaldehyde was added thereto, and the mixture was stirred overnight. Aqueous acetic acid was added, and the mixture was extracted with chloroform. The extract was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (eluent chloroform: methanol = 100: 1) to give 3-benzyl-7- [3- (Nt-butoxycarbonylamino) -1-pyrrolidinyl] -6-fluoro-1 , 4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine (325 mg) was obtained.
Melting point: 234-236 ° C

(2) 200 mg of the compound of the preceding item was dissolved in 15 ml of chloroform, 0.59 ml of trifluoroacetic acid was added under ice cooling, and the mixture was stirred for 1 hour and further stirred at room temperature overnight. The solvent was distilled off under reduced pressure, diisopropyl ether was added to the residue, and the precipitated crystals were collected by filtration to obtain 185 mg of the target compound (8F-3).
Melting point: 237-240 ° C

Example C-16 : --- Production of object (I)-:
7- (3-Amino-1-pyrrolidinyl) -3-formyl-1,4-dihydro-6-fluoro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine hydrochloride (Compound 8F-4) )

The title compound (8F-4) was obtained according to the method described in Example C-8.
Melting point: 242-250 ° C. (decomposition)

Example C-17 : --- Production of object (I)-:
7- (3-Amino-1-pyrrolidinyl) -6-fluoro-1,4-dihydro-3-hydroxy-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine trifluoroacetate (Compound 8F) -5)

(1) 7- [3- (Nt-butoxycarbonylamino) -1-pyrrolidinyl] -6-fluoro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine A mixture consisting of 0.5 g of -3-carboxylic acid, 0.75 g of potassium superoxide, 5 ml of ethanol and 5 ml of water was heated to reflux for 15 minutes, and the solvent was distilled off. The mixture was neutralized with aqueous acetic acid and extracted with chloroform. It dried with the anhydrous sodium sulfate and the solvent was depressurizingly distilled. The residue was recrystallized from chloroform-ethanol to give 7- [3- (Nt-butoxycarbonylamino) -1-pyrrolidinyl] -6-fluoro-1,4-dihydro-3-hydroxy-4-oxo-1- 260 mg of (2-thiazolyl) -1,8-naphthyridine was obtained.
Melting point: 251-252 ° C

(2) A mixture consisting of 260 mg of the compound of the preceding item, 6 ml of methylene chloride and 6 ml of trifluoroacetic acid was stirred at room temperature for 20 hours, and the solvent was distilled off under reduced pressure. Acetonitrile was added to the residue and the precipitated crystals were collected by filtration and washed with acetonitrile to obtain 170 mg of the target compound (8F-5).
Melting point: 206-207 ° C

Example C-18 : --- Production of object (I)-:
1,4-Dihydro-4-oxo-1- (2-thiazolyl) -7-vinyl-1,8-naphthyridine-3-carboxylic acid (Compound 9H-1)

(1) 7-chloro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester 3.6 g, vinyltributyltin 3.8 g, bistriphenylphosphine palladium dichloride A suspension consisting of 1.0 g and 150 ml of toluene was refluxed for 6.5 hours. After cooling with ice, it was washed with water, and the solvent was distilled off under reduced pressure. The obtained crystals were washed with toluene and ethanol to obtain 2.7 g of 1,4-dihydro-4-oxo-1- (2-thiazolyl) -7-vinyl-1,8-naphthyridine-3-carboxylic acid ethyl ester. It was.
Melting point: 163-164 ° C

(2) A suspension composed of 0.32 g of the compound of the preceding paragraph, 15 ml of 10% aqueous hydrochloric acid and 2 ml of ethanol was heated at 100 ° C. for 13 hours, and the crystals were collected by filtration and washed with ethanol to obtain the desired product 9H-1 (0.19 g )
Melting point: 284-288 ° C. (decomposition)

Example C-19 : --- Production of object (I)-:
1,4-Dihydro-7- (2,6-dimethyl-4-pyridinyl) -4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid (Compound 9H-2)

(1) 7-chloro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester 1.1 g, 2,6-dimethyl-4- (trimethyl) A suspension comprising 0.98 g of tin) pyridine, 0.25 g of bistriphenylphosphine palladium dichloride and 51 ml of toluene was refluxed for 6.5 hours. After cooling with ice, it was washed with water, and the solvent was distilled off under reduced pressure. Purification by silica gel column chromatography (eluent chloroform), washing with ethanol and 1,4-dihydro-7- (2,6-dimethyl-4-pyridinyl) -4-oxo-1- (2-thiazolyl)- 0.38 g of 1,8-naphthyridine-3-carboxylic acid ethyl ester was obtained.
Melting point: 232-235 ° C

(2) The target compound 9H-2 was obtained by treating in substantially the same manner as in Example C-18 (2).
Melting point: 272-275 ° C. (decomposition)

Example C-20 : --- Production of object (I)-:
7-ethynyl-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid (Compound 9H-3)

  (1) 7-chloro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester 1.5 g, (trimethylsilyl) ethynyltributyltin 2.0 g, bistriphenyl A suspension consisting of 0.45 g of phosphine palladium dichloride and 75 ml of toluene was heated at 100 ° C. under an argon atmosphere for 3 hours. After cooling with ice, it was washed with water, and the solvent was distilled off under reduced pressure. 1,4-dihydro-4-oxo-7- (trimethylsilyl) ethynyl-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester purified by silica gel column chromatography (eluent chloroform) 1.0 g was obtained.

(2) A suspension composed of 1.0 g of the compound obtained in the preceding paragraph, 0.45 g of potassium fluoride and 25 ml of ethanol was refluxed for 2 hours, and the solvent was distilled off under reduced pressure. Chloroform and water were added, insolubles were separated by celite filtration, washed with water, and the solvent was concentrated under reduced pressure. This was washed with a mixture of chloroform-ethanol to obtain 0.34 g of 7-ethynyl-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester. .
Melting point: 228-231 ° C. (decomposition)

(3) The compound obtained in the previous section was treated in substantially the same manner as in Example C-18 (2) to obtain the target compound 9H-3.
Melting point:> 300 ° C

Example C-21 : --- Production of object (I)-:
The following compounds were obtained by the method described in Example C-18 to Example C-20 or a method analogous thereto.

1,4-dihydro-4-oxo-7-phenyl-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid (compound 9H-4);
Melting point: 227-230 ° C (decomposition)

1,4-dihydro-7-methyl-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid (compound 9H-5);
Melting point: 268-269 ° C

1- (5-bromo-2-thiazolyl) -1,4-dihydro-7- (1-pyrrolidinyl) methyl-1,8-naphthyridine-3-carboxylic acid (compound 9H-6);
Melting point: 238-239 ° C

Example C-22 : --- Production of object (I)-:
1,4-dihydro-4-oxo-7- (3-pyrazolyl) -1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid (compound 9H-7)

(1) 7-ethynyl-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester 0.34 obtained in Example C-20 (2) g, a suspension composed of 1.3 g of trimethylsilyldiazomethane (10% hexane solution) and 20 ml of chloroform was refluxed for 5.5 hours. The solvent was distilled off under reduced pressure, washed with ethanol, and 1,4-dihydro-4-oxo-7- (3-pyrazolyl) -1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester 0.31 g was obtained.
Melting point: 285-287 ° C. (decomposition)

(2) The title compound 9H-7 was obtained by treating the compound obtained in the previous item in substantially the same manner as in Example C-18 (2).
Melting point:> 300 ° C

Example C-23 : --- Production of object (I)-:
6-Fluoro-1,4-dihydro-7-hydroxy-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid (Compound 9F-1)

To 10 g of 7-chloro-6-fluoro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester was added 10 ml of aqueous sodium hydroxide solution, 85 Stir at 6.5 ° C. for 6.5 hours. The reaction solution was acidified with aqueous hydrochloric acid, and the precipitated crystals were collected by filtration. The crystals were dissolved in a 5% aqueous sodium hydroxide solution, treated with activated carbon, acidified with hydrochloric acid, and the precipitated crystals were collected by filtration. The crystals were washed with water and then with ethanol to obtain 690 mg of the target compound.
Melting point: 261-264 ° C

Example C-24 : --- Production of object (I)-:
7- (3-Amino-1-pyrrolidinyl) -6-chloro-1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid 0.85 hydrochloride (compound Cl -1)

  (1) To a mixed solution consisting of acetic anhydride (60 ml) and concentrated nitric acid (5 ml), 2,6-dimethoxynicotinic acid (8.71 g) was added under ice cooling. After stirring at the same temperature for 3 hours and at room temperature for 4 hours, the precipitated crystals were collected by filtration and washed with isopropanol and diisopropyl ether. The filtrate was extracted with ethyl acetate, the solvent was distilled off under reduced pressure, and the resulting crystals were washed with isopropanol and diisopropyl ether, and the previous crystals were combined to give 2,6-dimethoxy-5-nitronicotinic acid (6.95 g). It was.

Melting point: 227-230 ° C. (decomposition);
IR (KBr) cm ⁻¹ ; 1706, 1610;
MS (m / z): 229 (MH ^<+> );
¹ H-NMR (DMSO-d ₆ ) δ: 4.03 (s, 3H), 4.08 (s, 3H), 8.71 (s, 1H)

  (2) Ethyl acetate (250 ml) was added to ethyl potassium malonate (18.6 g) and magnesium chloride (11.3 g), and triethylamine (28.0 g) was added dropwise to the resulting suspension under ice cooling. Stir for 2.5 hours.

  On the other hand, carbonyldiimidazole (14.8 g) was added to a solution of the compound (18.5 g) obtained in the previous section in tetrahydrofuran (150 ml), and the mixture was stirred at 70 ° C. for 1 hour. This solution was added to the previous suspension under ice cooling, and the mixture was stirred overnight at room temperature and further at 70 ° C. for 30 minutes. Concentrated hydrochloric acid was added to the reaction solution to make it acidic, and the precipitated crystals were collected by filtration and washed with ethyl acetate. The filtrate was extracted with ethyl acetate, the solvent was distilled off under reduced pressure, and the resulting crystals were recrystallized from a mixture of ethyl acetate and diisopropyl ether. Combined with the previous crystal, 2,6-dimethoxy-5-nitronicotinoyl acetic acid ethyl ester (19.9 g) was obtained.

MS (m / z): 299 (MH ^<+> );
¹ H-NMR (DMSO-d ₆ ) δ: 1.19 (t, 3H, J = 7 Hz), 4.00 (s, 2H), 4.10 (s, 3H), 4.11 (q, 2H, J = 7 Hz), 4.15 ( s, 3H), 8.78 (s, 1H)

  (3) A mixture consisting of the compound (2.2 g) obtained in the previous item, ethyl orthoformate (1.7 g) and acetic anhydride (2.4 g) was heated to reflux for 1.5 hours. The reaction mixture was concentrated under reduced pressure, and diisopropyl ether (10 ml) and 1,4-dioxane (3 ml) were added to the residue. To this solution was added 2-aminothiazole (0.84 g) under ice cooling, and the mixture was stirred overnight at room temperature. Precipitated crystals were collected by filtration and washed with diisopropyl ether to give 2- (2,6-dimethoxy-5-nitronicotinoyl) -3- (2-thiazolylamino) acrylic acid ethyl ester (2.3 g).

Melting point: 166-170 ° C;
IR (KBr) cm ⁻¹ ; 1729, 1603

  (4) Potassium carbonate (1.0 g) was added to a suspension composed of the compound (2.0 g) obtained in the previous section and 1,4-dioxane (90 ml), and heated at 60 ° C. for 3 hours. After cooling, water was added, neutralized with 20% aqueous hydrochloric acid, and the precipitated crystals were collected by filtration. Washing sequentially with water, ethanol and diisopropyl ether to give 1,4-dihydro-7-methoxy-6-nitro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid Ethyl ester (1.7 g) was obtained.

Melting point: 208-210 ° C;
IR (KBr) cm ⁻¹ ; 1739;
MS (m / z): 337 (MH ^<+> );
¹ H-NMR (DMSO-d ₆ ) δ: 1.31 (t, 3H, J = 7 Hz), 4.30 (s, 3H), 4.33 (q, 2H, J = 7 Hz), 7.85 (d, 1H, J = 4 Hz) ), 7.88 (d, 1H, J = 4 Hz), 9.08 (s, 1H), 9.69 (s, 1H)

(5) A dimethylformamide (150 ml) solution of the compound (10.3 g), 3- (t-butoxycarbonylamino) pyrrolidine (5.9 g) and potassium carbonate (4.7 g) obtained in the previous section was heated at 60 ° C. for 2 hours. . After cooling, water was added and extracted with ethyl acetate. The extract was washed with water and saturated brine, and dried over sodium sulfate. The residue obtained by distilling off the solvent under reduced pressure was recrystallized from a mixture of ethyl acetate and ethanol to give 7- (3-t-butoxycarbonylamino-1-pyrrolidinyl) -1,4-dihydro-6-nitro- 4-Oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester (11.4 g) was obtained.
Melting point: 204-205 ° C

(6) Dissolve the compound (1.0 g) obtained in the previous section in ethanol (100 ml) and 1,4-dioxane (50 ml), add Raney-nickel (about 1 g) and absorb hydrogen at 50 ° C. for 2.5 hours. It was. Precipitated crystals were collected by filtration, dimethylformamide was added and heated, and insolubles were filtered off. The filtrate was concentrated under reduced pressure, and the resulting crystals were washed with a mixed solution consisting of dimethylformamide, 1,4-dioxane and ethanol to give 6-amino-7- (3-tert-butoxycarbonylamino-1-pyrrolidinyl) -1, 4-Dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester (0.81 g) was obtained.
Melting point: 270-272 ° C. (decomposition)

(7) A solution of sodium nitrite (60 mg) in water (2 ml) was added dropwise to a mixture of the compound obtained in the previous item (250 mg) and concentrated hydrochloric acid (5 ml) under ice cooling, and the mixture was stirred at the same temperature for 10 minutes. The reaction solution was added dropwise to a mixture of copper (I) chloride (70 mg) and concentrated hydrochloric acid (2 ml) under ice cooling. The mixture was stirred at the same temperature for 1 hour and at 60 ° C. for 30 minutes, made alkaline with a 20% aqueous sodium hydroxide solution, and then extracted with chloroform. The extract was dried over sodium sulfate, the solvent was distilled off under reduced pressure, and the resulting residue was recrystallized from a mixture of chloroform and diisopropyl ether to give 7- (3-amino-1-pyrrolidinyl) -6-chloro-1. , 4-Dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid ethyl ester (30 mg) was obtained.
Melting point: 208-210 ° C

(8) The desired 7- (3-amino-1-pyrrolidinyl) -6-chloro-1 by hydrolyzing the ethyl ester obtained in the previous item according to the method described in Example C-1 (2) , 4-Dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid 0.85 hydrochloride (compound Cl-1) was obtained.
Melting point: 288-291 ° C (decomposition)

Example C-25 : --- Production of object (I)-:
The following compounds were obtained by the method described in any of the C-series examples or a method analogous thereto.

    Table 22

Example C-26 : --- Production of object (I)-:
3- [1,4-Dihydro-7- (trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2-thiazolyl) -1,8-naphthyridin-3-yl] Acrylic acid methyl ester 0.9 Trifluoroacetate (Compound 9H-9)

  3-Formyl-1,4-dihydro-7- (trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2-thiazolyl) -1 obtained in Example C-8 , 8-Naphthyridine hydrochloride (100 mg) was added 3 ml of aqueous ammonia and extracted with chloroform. The extract was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to give an oil.

  On the other hand, 60% sodium hydride (19 mg) was washed with n-hexane, and 10 ml of tetrahydrofuran was added. Trimethylphosphonoacetate (65 mg) was added to the resulting suspension at room temperature, stirred for 10 minutes, then ice-cooled, and a solution of the oil obtained above in tetrahydrofuran (5 ml) was added dropwise. After stirring at room temperature for 3 hours, water was added and the mixture was extracted with chloroform. The extract was washed with saturated brine and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent chloroform: methanol = 50: 1) to give an oil. This was dissolved in methylene chloride, 5 drops of trifluoroacetic acid was added under ice cooling, diisopropyl ether was further added, and the precipitated crystals were collected by filtration to obtain the title compound (22 mg).

Melting point: 230-233 ° C. (decomposition);
IR (KBr) cm ⁻¹ ; 2950, 1680, 1620;
MS (m / z): 442 (MH ^<+> )

Example C-27 : --- Production of object (I)-:
3-acetyl-1,4-dihydro-7- (trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2-thiazolyl) -1,8-naphthyridine (Compound 9H- 10)

  (1) Ethyl acetate (60 ml) was added to ethyl potassium malonate (572 mg), and magnesium chloride (343 mg) and triethylamine (1.0 ml) were added to the resulting suspension at room temperature and stirred for 5 hours.

  On the other hand, 7- [trans-3- (Nt-butoxycarbonylmethylamino) -4-methoxy-1-pyrrolidinyl] -1,4-dihydro-4-oxo- obtained in Example C-8 (2) To a solution of 1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid (1.2 g) in chloroform (120 ml), BOP reagent (1.27 g) and triethylamine (0.8 ml) were added at room temperature, and the mixture was stirred for 20 minutes. did. This reaction solution was added to the previous reaction solution at room temperature and stirred for 1 day.

The reaction solution was poured into water and extracted with chloroform. The extract was washed with saturated brine and dried over anhydrous sodium sulfate. The residue obtained by evaporating the solvent under reduced pressure was purified by silica gel column chromatography (eluent chloroform: methanol = 200: 1) to give 3- {7- [trans-3- (Nt-butoxycarbonylmethyl]. Amino) -4-methoxy-1-pyrrolidinyl] -1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridin-3-yl} -3-oxopropionic acid ethyl ester (397 mg).
MS (m / z): 572 (MH ^<+> )

  (2) The compound (170 mg) obtained in (1) above was dissolved in ethanol (2 ml), and 10% aqueous hydrochloric acid (10 ml) was added. The mixture was stirred at room temperature for 30 minutes and at 95 ° C. for 4 hours. After cooling, the solution was made alkaline with aqueous sodium hydrogen carbonate, and then extracted with chloroform. The extract was washed with saturated brine and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was crystallized from diisopropyl ether to obtain the title compound (76 mg).

Melting point: 112-114 ° C;
IR (KBr) cm ⁻¹ ; 3085, 1685, 1620;
MS (m / z): 400 (MH ^<+> )

Example C-28 : --- Production of object (I)-:
N- [1,4-dihydro-7- (trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carbonyl] Glycine ethyl ester trifluoroacetate (Compound 9H-11)

  According to the method described in Example C-11 (1), 7- [trans-3- (Nt-butoxycarbonylmethylamino) -4-methoxy-1-pyrrolidinyl] -1,4-dihydro-4 -Oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid (250 mg) and hydrochloric acid glycine ethyl ester (78 mg) were converted to N- {7- [trans-3- (N-t-butoxycarbonyl]. Methylamino) -4-methoxy-1-pyrrolidinyl] -1,4-dihydro-4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carbonyl] glycine ethyl ester (290 mg) was obtained. .

This was dissolved in methylene chloride (9 ml), and trifluoroacetic acid (1.2 ml) was added under ice cooling. After stirring at room temperature for one day, diisopropyl ether was added to the reaction mixture, and the precipitated crystals were collected by filtration to obtain the desired compound 9H-11 (260 mg).
Melting point: 233-236 ° C. (decomposition)

Example C-29 : --- Production of object (I)-:
N- [1,4-dihydro-7- (trans-3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carbonyl] -L-alanine trifluoroacetate (Compound 9H-12)

The title compound was obtained in the same manner as in Example C-28.
Melting point: 168-170 ° C (decomposition)

  Next, preparation examples of preparations are given.

Example D-1 :-Preparation of solution--:
Formula:-

  Preparation method: Compound H-1-1-1 and sorbitol were dissolved in a part of distilled water for injection, sodium hydroxide and the remaining distilled water were added, and the pH of the solution was adjusted to 5.0. This solution was filtered through a membrane filter (0.22 μm) to obtain an injection solution.

Example D-2 :-Preparation of lyophilized formulation--:
Formula:-

  Preparation method: Compound H-1-1-1 and mannitol were dissolved in a part of distilled water for injection, sodium hydroxide and the remaining distilled water were added, and the pH of the solution was adjusted to 5.0. This solution was filtered through a membrane filter (0.22 μm) and freeze-dried to obtain a powder for injection.

  The compound according to the present invention is not limited to non-solid tumors such as lymphocytic leukemia tumors, but also to various solid tumors occurring in tissues such as lung, milk, stomach, uterus, skin, intestine, bladder, and nasal throat. It also shows a marked antitumor effect. In addition, the compound according to the present invention has high safety for living bodies. Therefore, the compound according to the present invention is useful as a therapeutic or prophylactic agent for human tumors.

It is a figure which shows the anti-tumor effect with respect to human nasopharyngeal carcinoma (KB). It is a figure which shows the anti-tumor effect with respect to human breast cancer (MX-1). It is a figure which shows the anti-tumor effect with respect to a human colon cancer (WiDr). These are figures which show the anti-tumor effect with respect to human melanoma (HMV-2). It is a figure which shows the anti-tumor effect with respect to human lung cancer (LX-1).

Claims (4)

The compound represented by the following general formula (I), its stereoisomer or optical isomer, or a physiologically acceptable salt thereof, or any one of the hydrates or solvates described above as an active ingredient Tumor agents:
(In the formula, --------- means the presence or absence of a single bond. X means C-Rx, where Rx means a hydrogen atom. R ₁ and R ₂ are The same or different, a hydrogen atom, a halogen atom, a nitro group, a lower alkoxy group, a lower alkyl group (the lower alkyl group may be substituted with lower alkoxycarbonyl or carboxyl), or a phenyl group optionally substituted with halogen Or a thienyl group, or a phenylsulfonyl group (wherein the phenyl moiety may be substituted with halogen or nitro), or R ₁ and R ₂ together form a benzene ring or a naphthalene ring, a benzene ring or a naphthalene ring which may be substituted by halogen, nitro or lower alkyl .R ₃ is a hydrogen atom, a benzyl group, a hydroxyl group, carbo Sill group, a hydroxymethyl group, .R ₄ to mean a formyl group or a lower alkyl ester is .R ₅ to mean an oxo group or a hydroxy group and a hydrogen atom, an amino group, a halogen atom or a lower alkyl group, lower alkyl The group may be substituted with halogen, and R ₆ represents any group selected from the following (a) to (e).
(A) A 4- to 7-membered monocyclic amino group represented by the following formula (a):

(Wherein --------- means the presence or absence of a single bond, R6a1, R6a2, R6a3 and R6a4 are identical or _{different, C, CH, CH 2,} CH 2 CH 2, CH ═CH, S (O) _0-2 , O, N or N—R 6a, wherein R 6a represents a hydrogen atom, a lower alkyl group or a phenyl group, and the phenyl group represents lower alkoxy, lower alkyl, halogen Or k and m are the same or different and each represents an integer of 0 or 1. R6a5 represents a hydrogen atom, a methylene group or a group represented by the following formula (R6a-5) To:

[Wherein R6a51 represents a hydrogen atom or a phenyl group (the phenyl group may be substituted with halogen), or a lower alkyl group optionally substituted with phenyl or halogenophenyl; Is a hydrogen atom, a lower alkyl group or a group that can be converted to a hydrogen atom, and n is an integer of 0-4. ]
R6a6 means a hydrogen atom, a halogen atom, a hydroxyl group, an azido group, a formyl group, an oxo group, a lower alkoxy group, a lower alkylthio group or a lower alkyl group (the lower alkyl group may be substituted with halogen or hydroxy). . R6a7 means the same as R6a6 or a spirocyclic amino group represented by the following formula (R6a-7):
[Wherein p represents an integer of 3 or 4. ]
(B) a bicyclic amino group represented by the following formula (b):

(Where --------- means the presence or absence of a single bond, q, s and t are the same or different and are integers from 0 to 2, and the sum of q and t is 2) or 3 .R6b1 means C, CH, a CH ₂ or N-R6b, wherein the R6b is a hydrogen atom or a lower alkyl group, R6b2 is C, CH, CH _2, CH ₂ CH ₂ or CH ═CH, R 6b3 means C, CH, CH ₂ or O, u, v and w are the same or different and are integers of 0 or 1, and their sum is 1 to 3 R6b4 and R6b5 are the same or different and each represents a hydrogen atom, a hydroxyl group, a lower alkyl group, an amino group, a mono- or di-lower alkylamino group, or a mono- or di-lower alkylamino lower alkyl group.
(C) A 3- to 7-membered cyclic group represented by the following formula (c):

(Wherein --------- means the presence or absence of a single bond, R6c1, R6c2, R6c3 and R6c4 are identical or _{different, C, CH, CH 2,} CH 2 CH 2, CH ═CH, S (O) _0-2 , O, N or N—R6c, wherein R6c represents a hydrogen atom or a lower alkyl group, R6c5 represents O, S or CH ₂ , R6c6 and R6c7 is the same or different and represents a hydrogen atom, an amino group or a lower alkyl group, and f, g and h are the same or different and represent an integer of 0 or 1.)
(D) Amino group represented by the following formula (d):

(Wherein R6d1 means S or NH, R6d2 and R6d3 are the same or different and each represents a hydrogen atom or a lower alkyl group, i represents an integer of 0 or 1, and j represents an integer of 1 to 5) means.)
(E) a hydroxyl group, a lower alkyl group, a lower alkenyl group or a lower alkynyl group. However, 1,4-dihydro-7- (3-methoxy-4-methylamino-1-pyrrolidinyl) -4-oxo-1- (2-thiazolyl) -1,8-naphthyridine-3-carboxylic acid, its steric form Excludes isomers or optical isomers or physiologically acceptable salts thereof. ) A compound represented by the following general formula (Ia), a stereoisomer or optical isomer thereof, or a physiologically acceptable salt thereof, or any of the hydrates or solvates described above as an active ingredient The antitumor agent according to claim 1, wherein

(Wherein -----, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , X, R 6a ₁ , R 6a ₂ , R 6a ₃ , R 6a ₄ , R 6a ₅ , R 6a ₆ , R 6a 7 and k are defined in claim 1) Means the same as described.) A compound represented by the following general formula (Ib), a stereoisomer or optical isomer thereof, or a physiologically acceptable salt thereof, or any one of the hydrates or solvates described above as an active ingredient The antitumor agent according to claim 2, wherein:
Wherein W is a hydrogen atom, and R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R 6a ₅ , R 6a 6 and R 6a 7 are the same as described in claim 1. Means things.) A compound represented by the following general formula (Ic), a stereoisomer or optical isomer thereof, or a physiologically acceptable salt thereof, or any of the hydrates or solvates described above as an active ingredient The antitumor agent according to claim 3, wherein
(Wherein R6a511 is a lower alkyl group, and R6a61 and R6a71 are the same or different and each represents a hydrogen atom, a lower alkoxy group or a lower alkyl group.)