JP2011246362A - Alaremycin derivative - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a new alaremycin derivative that has inhibitory activity against porphobilinogen synthase (PBGS) and is useful as an antibacterial agent.SOLUTION: The alaremycin derivative is represented by formula (I) (wherein R is a hydrogen atom or lower alkyl; X is halogen-substituted methyl; when one of Yand Yis a hydrogen atom, the other is a hydrogen atom, hydroxymethyl or lower alkoxymethyl or Yand Yare bonded to form a double bond; m is an integer of 0-5) or its pharmacologically acceptable salt.

Description

  The present invention relates to a novel araremycin derivative having an inhibitory activity on porphobilinogen synthase (PBGS).

  Recently, for example, methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VER) and multi-drug resistant Pseudomonas aeruginosa (MDRP) having resistance to existing antibiotics Is spreading throughout the world and has become a major problem, including nosocomial infections. The development of new antibacterial agents having antibacterial activity against these multidrug-resistant bacteria has always been desired.

  In 2005, Awa et al. Performed antibacterial activity against Escherichia coli from a medium in which Streptomyces sp. A012304 was grown by screening using nonnucleated cells released as a result of inhibition of chromosome distribution (Non-patent Document 1). Is a novel structure antibiotic having the following formula (A):

"Araremycin" represented by (Non-Patent Document 2).

Aralemycin acts as an inhibitor of PBGS, a porphyrin-heme biosynthetic enzyme that synthesizes porphobilinogen (PBG) using 5-aminolevulinic acid (5-ALA) as a substrate. (Non-patent Document 3). PBGS is classified into two types based on the difference in metal ions that are cofactors. One is a Zn ²⁺ type distributed in animal cells including humans and many bacteria, and the other is one such as plant cells and Pseudomonas aeruginosa. Mg ²⁺ type distributed in some bacteria (Non-Patent Documents 4, 5, and 6).

PBGS is involved in the synthesis of porphyrins and related compounds that are indispensable for the survival of living organisms. Actually, PBGS inhibitor araremycin has antibacterial activity against E. coli, and its structure differs from humans. Pseudomonas aeruginosa with Mg ²⁺ type PBGS is a pathogen that has become a problem as a multi-drug resistant bacterium that is opportunistically infected in hospitals. From these, Mg ²⁺ type PBGS is effective in terms of antibacterial agents. It can be a target molecule.

However, in the drug sensitivity test conducted in the report of Heinemann et al., The IC ₅₀ (50% inhibitory concentration) of P. aeruginosa PBGS by araremycin is 2.1 mM, so that it can be used as an antibacterial agent. Has been required to enhance antibacterial activity (Non-patent Document 3).

  Also known as porphyria is a group of diseases caused by abnormal accumulation of intermediary metabolite porphyrin or its precursor due to genetic failure of any of the eight enzymes involved in the heme biosynthetic pathway. It has been. Clinically, skin characterized by acute porphyria with symptoms such as gastrointestinal symptoms (abdominal pain, vomiting, constipation, etc.) and neurological symptoms (motor paralysis and limb sensory impairment) and skin symptoms (photosensitivity) It can be broadly classified into type porphyria.

  Among them, erythroblastic protoporphyria (EPP), which develops when the skin is irradiated with sunlight, may be fatal due to liver failure at a young age. However, porphyria currently does not have a fundamental cure at all, and the burden on patients and their families, including the problem of being a genetic disease, is very large. Furthermore, there are few medical institutions that specialize in the treatment of porphyria, and in this respect it cannot be said that patients are receiving sufficient follow-up.

  An acid amide compound of araremycin represented by the following formula (B) is known as Primocarcin, an anticancer antibiotic (Non-patent Document 7). Further, a methyl ester compound of araremycin represented by the following formula (C) is known as a photosensitizer for cancer photodynamic therapy (Patent Document 1).

JP 2006-282577 A

Biochimie, 1999, 81, 909-913. Biosci. Biotechnol. Biochem. 2005, 69, 1721-1725. Antimicrob. Agents Chemother. 2010, 54, 267-272. J. et al. Bioenerg. Biomembr. 1995, 27, 169-179. Biochem. J. et al. 1996, 320, 401-412. Mol. Gen. Genet. 1998, 257, 485-489. J. et al. Antibiotics, Ser. A, 15, 77-79

  An object of the present invention is to provide a novel araremycin derivative having excellent PBGS inhibitory activity.

  The present inventors have found that a compound having an excellent PBGS inhibitory effect can be obtained by changing the methyl group of the acetyl group of araremycin to a methyl group substituted with a halogen atom, and the present invention has been completed. .

That is, the present invention
(1) Formula (I)

(In the formula, R represents a hydrogen atom or a lower alkyl group, X represents a halogen-substituted methyl group, Y ¹ and Y ² represent a hydrogen atom when one represents a hydrogen atom, and the other represents a hydrogen atom or a hydroxymethyl group. Or a lower alkoxymethyl group, or Y ¹ and Y ² together represent a double bond, and m represents an integer of 0 to 5).

The present invention relates to an araremycin derivative represented by the formula: or a pharmacologically acceptable salt thereof.

The present invention also provides:
(2) The compound represented by the formula (I) is represented by the following formula (Ia)

(Wherein R, X and m are as defined above)

An aralemycin derivative or a pharmacologically acceptable salt thereof according to the above (1), which is represented by:
(3) The araremycin derivative or the pharmacologically acceptable salt thereof according to the above (2), wherein R is a hydrogen atom,
(4) m is an integer of 0 to 2, the araremycin derivative according to any one of the above (1) to (3) or a pharmacologically acceptable salt thereof,
(5) X is a fluorine-substituted methyl group, the aralemycin derivative according to any one of (1) to (4) above, or a pharmacologically acceptable salt thereof,
(6) The araremycin derivative or the pharmaceutically acceptable salt thereof according to (5), wherein the fluorine-substituted methyl group is a trifluoromethyl group.

The present invention also provides:
(7) Inhibitor of porphobilinogen synthase (PBGS) containing the araremycin derivative or the pharmacologically acceptable salt thereof according to any one of (1) to (6) as an active ingredient About.

Furthermore, the present invention provides
(8) The present invention relates to an antibacterial agent containing the araremycin derivative or the pharmacologically acceptable salt thereof according to any one of (1) to (6) as an active ingredient.

  The novel araremycin derivative of the present invention has an excellent PBGS inhibitory activity and is useful as a therapeutic agent for infectious diseases, particularly as an antibacterial agent against multidrug-resistant bacteria.

  Specific examples of the definition of each group in the compound (I) are shown below, but these are preferable examples of the present invention, and of course not limited thereto.

  The lower alkyl group is, for example, linear or branched alkyl having 1 to 6 carbon atoms, specifically, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, Neopentyl, hexyl and the like can be mentioned.

  Examples of the halogen atom as the substituent of the halogen-substituted methyl group include fluorine, chlorine, bromine and iodine atoms. The number of substituents is the same or different and is 1 to 3. The halogen-substituted methyl group is not particularly limited as long as the hydrogen atom of the methyl group is substituted with a halogen atom, but is preferably a group substituted with a fluorine atom, such as monofluoromethyl, difluoromethyl, trifluoromethyl. And a trifluoromethyl group is particularly preferable.

  The alkyl part of the lower alkoxymethyl group has the same meaning as the above alkyl group. For example, the linear or branched alkoxymethyl having 2 to 7 carbon atoms, specifically, methoxymethyl, ethoxymethyl, propoxymethyl, isopropoxy Examples include methyl, butoxymethyl, tert-butoxymethyl, pentyloxymethyl, hexyloxymethyl and the like.

  Examples of the pharmacologically acceptable salt of the compound represented by the formula (I) include pharmacologically acceptable metal salts, ammonium salts, organic amine addition salts and the like, and are pharmacologically acceptable. As the metal salt, each alkali metal salt such as lithium, sodium and potassium, each alkaline earth metal salt such as magnesium and calcium, each metal salt such as aluminum and zinc are pharmaceutically acceptable ammonium salts. Organic salts that are pharmacologically acceptable for salts such as ammonium and tetramethylammonium include salts such as triethylamine, piperidine, morpholine, and toluidine.

  Next, the manufacturing method of compound (I) is demonstrated.

Manufacturing method 1.
The compound (Ia) in which Y ¹ and Y ² are combined to form a double bond, and the compound (Ib) in which Y ¹ is a hydrogen atom and Y ² is hydroxymethyl or lower alkoxymethyl are prepared according to the following reaction steps: Can be manufactured.

(Wherein Q represents an amino-protecting group, Y ³ represents a lower alkyl group, Hal represents a halogen atom, and R, X and m are as defined above).

(Process a)
The amino group of serine methyl ester compound (II) is protected by a conventional method. Any protecting group may be used as long as it is used as an amino group protecting group for amino acids, but a tert-butoxycarbonyl (Boc) group is preferably used in consideration of the subsequent reaction steps.

(Process b)
Compound (III) can be obtained by alkylating compound (II) by a conventional method.

(Process c)
The aldehyde compound (IV) can be obtained by reducing the methyl ester of the compound (III). The reducing agent is not particularly limited as long as it is a reducing agent that reduces an ester to an aldehyde, but diisobutylaluminum hydride (DIBAL-H) is preferably used.

(Process d)
Compound (VI) comprises compound (IV) and acrylate compound (V), a thiazole catalyst such as 3-ethyl-5- (2-hydroxyethyl) -4-methylthiazolium bromide and diazabicyclo It can be obtained by reacting in the presence of a base such as undecene (DBU).

(Process e)
Compound (VII) can be obtained by deprotecting the amino-protecting group of compound (VI) by a conventional method. For example, when Q is a Boc group, compound (VII) can be obtained by treating compound (VI) with trifluoroacetic acid.

(Process f)
The compound (Ib) of the present invention in which Y ¹ is a hydrogen atom and Y ² is lower alkoxymethyl can be obtained by reacting the compound (VII) with an acid halogen compound (VIII) in the presence of a base such as triethylamine. .

When the compound (Ib) in which Y ² is hydroxymethyl (Y ³ = H) is desired, it can be obtained by hydrolyzing the compound (Ib) in which Y ² obtained above is lower alkoxymethyl. it can. For example, compound (Ib) in which Y ² is hydroxymethyl can be obtained by treating compound (Ib) in which Y ² is tert-butoxymethyl (Y ³ = t-Bu) with trifluoroacetic acid.

(Process g)
The compound (Ia) of the present invention in which Y ¹ and Y ² are combined to form a double bond is a compound (Ib) in which Y ¹ obtained above is a hydrogen atom and Y ² is hydroxymethyl (Y ³ = H). ) Can be obtained by subjecting to a dealcoholization reaction. For example, the compound (Ib) in which Y ³ = H is reacted in the presence of a Boc agent such as di-tert-butyl dicarbonate (Boc ₂ O) and a base such as dimethylaminopyridine (DMAP), followed by It can be obtained by treatment with a base such as 1,1,3,3-tetramethylguanidine (TMG).

  Further, when a compound in which R is a hydrogen atom is desired among compounds (Ia) and (Ib), the ester compound (Ia) or (Ib) in which R is a lower alkyl group is hydrolyzed by a conventional method. Can be obtained.

Production method 2.
In compound (Ia), a compound in which m is 0 can also be produced according to the following reaction step.

(Wherein R and X are as defined above)

(Process h)
Compound (Ia) in which m is 0 is obtained by subjecting azide compound (IX) to sodium perrhenate (NaReO ₄ ) and trifluoromethanesulfonic acid (CF) according to the method described in the literature (Synlett, 2006, 3, 481-483). It can be obtained by reacting the acid anhydride compound (X) using ₃ SO ₃ H) as a catalyst.

Production method 3.
Compound (Ic) in which Y ¹ and Y ² are both hydrogen atoms can be produced according to the following reaction step.

(Wherein R, X, Q, Hal and m are as defined above)

(Steps i to l)
The compound (Ic) of the present invention in which Y ¹ and Y ² are both hydrogen atoms can be obtained from the glycine methyl ester (XI) in which the amino group is protected according to the methods of steps cf.

  The reaction in each of the above steps a to l is a solvent inert to an appropriate reaction, for example, a halogenated hydrocarbon such as chloroform and dichloromethane, an aromatic hydrocarbon such as benzene and toluene, diethyl ether, tetrahydrofuran (THF), Ether solvents such as 1,4-dioxane, aprotic polar solvents such as N, N-dimethylformamide (DMF), N-methylpyrrolidone (NMP) and dimethyl sulfoxide (DMSO), and basic solvents such as pyridine and quinoline And in water or a mixed solvent thereof at a temperature between −78 ° C. and the boiling point of the solvent used for 5 minutes to 48 hours.

  In each of the above production methods, when the defined group changes under the conditions of the method of implementation or is inappropriate for carrying out the method, the method for introducing and removing protecting groups commonly used in organic synthetic chemistry should be used. The target compound can be obtained. Moreover, the conversion of the functional group contained in each substituent can be carried out by a known method other than the above production method, and in compound (I), another derivative ( Some can lead to I).

  The intermediates and target compounds in each of the above production methods are isolated and purified by purification methods commonly used in synthetic organic chemistry, such as neutralization, filtration, extraction, washing, drying, concentration, recrystallization, and various chromatography. can do. In addition, the intermediate can be subjected to the next reaction without any particular purification.

  Some compounds (I) may have isomers such as optical isomers, but the present invention includes all possible isomers and mixtures thereof.

  When it is desired to obtain a salt of compound (I), if compound (I) is obtained in the form of a salt, it can be purified as it is, and if it is obtained in a free form, it can be dissolved in an appropriate organic solvent. Alternatively, it may be suspended and an acid added to form a salt by a conventional method.

  Compound (I) and pharmacologically acceptable salts thereof may exist in the form of adducts with water or various solvents, and these adducts are also included in the present invention.

  Specific examples of the compound (I) obtained by the above production method are shown in Tables 1 to 3.

  The compound of the present invention represented by the formula (I) has PBGS inhibitory activity, is useful as an antibacterial agent, and the compound (I) that can be used as a PBGS inhibitor is compound (I). Although not particularly limited, it is preferably compound (Ia) or compound (Ic), more preferably compound (Ia), compound (Ia-2), (Ia-4), or (Ia-5). Is particularly preferred. Compound (Ib) is also useful as a synthetic intermediate for compound (Ia).

  The compound (I) or a pharmacologically acceptable salt thereof can be administered alone as it is, but it is usually desirable to prepare various pharmaceutical preparations. It can be produced by a conventional method of pharmaceutics by mixing with one or two or more types of carriers that are acceptable.

  Examples of the administration route include oral administration, inhalation administration, and parenteral administration such as intravenous administration.

  Examples of the dosage form include tablets, injections, etc. The tablets are mixed with various additives such as lactose, starch, magnesium stearate, hydroxypropyl cellulose, polyvinyl alcohol, surfactant, glycerin, etc. The inhalant may be produced according to a conventional method by adding, for example, lactose. An injection may be produced according to a conventional method by adding water, physiological saline, vegetable oil, solubilizer, preservative and the like.

  The effective amount and frequency of administration of compound (I) or a pharmacologically acceptable salt thereof vary depending on the administration form, patient age, body weight, symptoms, etc., but usually 0.001 mg to 5 g per adult, preferably Is administered in a dose of 0.1 mg to 1 g, more preferably 1 to 500 mg, once a day or several times a day.

  EXAMPLES Hereinafter, although an Example demonstrates this invention more concretely, the technical scope of this invention is not limited to these illustrations.

4-Oxo-5- (trifluoroacetylamino) -5-hexenoic acid methyl compound (Compound Ia-1)
A solution of methyl 5-azido-4-oxohexanoate (93 mg, 0.50 mmol) and sodium perrhenate (2 mg, 7.5 μmol) in trifluoroacetic anhydride (1.0 ml) and carbon tetrachloride (1.0 ml). Trifluoromethanesulfonic acid (1.8 μl, 20 μmol) was added with stirring, and the mixture was heated to reflux for 12 hours with stirring. Subsequently, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 75 mg (yield 60%) of the title compound (Ia-1).

¹ H NMR (CDCl ₃ ) δ 2.72 (t, J = 6.4 Hz, 2H), 3.13 (t, J = 6.4 Hz, 2H), 3.72 (s, 3H), 6.10 (t, J = 1.7 Hz, 1H) , 7.03 (d, J = 1.7 Hz, 1H), 8.85 (s, 1H).
¹⁹ F NMR (CDCl ₃ ) δ 85.58 (s, 3F).
¹³ C NMR (CDCl ₃ ) δ 27.7, 30.5, 51.9, 112.5, 115.2 (q, J = 288.6 Hz), 136.3, 155.2 (q, J = 37.9 Hz), 172.4, 193.8.

4-Oxo-5- (trifluoroacetylamino) -5-hexenoic acid (Compound Ia-2)
While stirring a solution of the compound (Ia-1) obtained in Example 1 (65 mg, 0.26 mmol) in THF (1.0 ml) and water (1.0 ml), 1N lithium hydroxide (0.6 ml) was added. Added at 0 ° C. After stirring for 1 hour at 0 ° C., the reaction mixture was adjusted to pH = 3 with 10% tartaric acid and extracted three times with ethyl acetate. The organic layers were combined and dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 30 mg (yield 50%) of the title compound (Ia-2).

¹ H NMR (CDCl ₃ ) δ 2.77 (t, J = 6.4 Hz, 2H), 3.13 (t, J = 6.4 Hz, 2H), 6.09 (t, J = 1.7 Hz, 1H), 7.04 (d, J = 1.7 Hz, 1H), 8.85 (s, 1H).
¹⁹ F NMR (CDCl ₃ ) δ 85.58 (s, 3F).
¹³ C NMR (CDCl ₃ ) δ 27.6, 30.2, 112.6, 115.2 (q, J = 288.7 Hz), 136.2, 155.2 (q, J = 37.9 Hz), 177.6, 193.5.

Methyl 6- (tert-butoxy) -4-oxo-5-[(3,3,3-trifluoro-1-oxopropyl) amino] hexanoate (Compound Ib-1)

(1) Methyl 2- (tert-butoxycarbonylamino) -3-hydroxypropionate (III-1: Q = Boc, Y ³ = H)
While stirring a solution of serine methyl ester hydrochloride (1.6 g, 10 mmol) and triethylamine (3.1 ml, 22 mmol) in anhydrous methylene chloride (20 ml), (Boc) ₂ O (2.5 ml, 11 mmol) was added at 0 ° C. Slowly added. The mixed solution was stirred at 0 ° C. for 30 minutes, and further stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure and partitioned between ethyl acetate and 1M aqueous KHSO ₄ solution. The organic layer was separated, washed successively with 1M KHSO ₄ aqueous solution, saturated multistory water, and brine, and then dried over anhydrous magnesium sulfate. Then, the solvent was distilled off under reduced pressure to obtain 2.0 g (yield 92%) of the title compound (III-1).

¹ H NMR (CDCl ₃ ) δ 1.45 (s, 9H), 3.05 (br s, 1H), 3.78 (s, 3H), 3.86-3.91 (m, 1H), 3.94-3.99 (m, 1H), 4.38 ( br s, 1H), 5.60 (d, J = 7.3 Hz, 1H).
¹³ C NMR (CDCl ₃ ) δ 28.2, 52.3, 55.7, 63.2, 80.2, 155.6, 171.2.

(2) Methyl 3- (tert-butoxy) -2- (tert-butoxycarbonylamino) propionate (III-2: Q = Boc, Y ³ = ^t Bu)
While stirring a solution of compound (III-1) obtained in (1) (13.2 g, 60 mmol) and Mg (ClO ₄ ) ₂ (1.3 g, 6.0 mmol) in anhydrous methylene chloride (200 ml), ( Boc) ₂ O (30.3 ml, 132 mmol) was added and the mixture was stirred at room temperature for 12 hours. The reaction solution was diluted with water and extracted with methylene chloride. The extract was dried over anhydrous magnesium sulfate and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography to obtain 8.6 g (yield 52%) of the title compound (III-2).

¹ H NMR (CDCl ₃ ) δ 1.13 (s, 9H), 1.46 (s, 9H), 3.56 (dd, J = 9.0, 3.2 Hz, 1H), 3.74 (s, 3H), 3.79 (dd, J = 9.0 , 3.0 Hz, 1H), 4.39 (ddd, J = 8.1, 3.2, 3.0 Hz, 1H), 5.36 (d, J = 8.1 Hz, 1H).
¹³ C NMR (CDCl ₃ ) δ 27.2, 28.3, 52.0, 54.3, 62.1, 73.2, 79.7, 155.5, 171.3.

(3) 3- (tert-butoxy) -2- (tert-butoxycarbonylamino) propanal (IV-1: Q = Boc, Y ³ = ^t Bu)
While stirring a solution of compound (III-2) (8.1 g, 29 mmol) obtained in (2) above in anhydrous toluene (40 ml), DIBAL-H (1.0 M toluene solution; 38 ml, 38 mmol) was added at −78 ° C. Was gradually added. The reaction solution was stirred at −78 ° C. for 2 hours, and then the excess DIBAL-H was treated with 3N hydrochloric acid aqueous solution (20 ml) to stop the reaction. Subsequently, the reaction solution was filtered, and the organic layer of the filtrate was washed with saturated multistory water, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography to obtain 6.4 g (yield 89%) of the title compound (IV-1).

¹ H NMR (CDCl ₃ ) δ 1.15 (s, 9H), 1.47 (s, 9H), 3.60 (dd, J = 9.3, 3.9 Hz, 1H), 3.92 (dd, J = 9.3, 3.1 Hz, 1H), 4.25 (ddd, J = 7.0, 3.9, 3.1 Hz, 1H), 5.38 (d, J = 7.0 Hz, 1H), 9.62 (s, 1H). ¹³ C NMR (CDCl ₃ ) δ 27.1, 28.2, 60.1, 73.5 , 79.9, 155.6, 199.7.

(4) Methyl 6- (tert-butoxy) -5- (tert-butoxycarbonylamino) -4-oxohexanoate (VI-1: R = Me, Q = Boc, Y ³ = ^t Bu)
Compound (IV-1) (0.49 g, 2.0 mmol) obtained in the above (3), molecular sieve (MS-4A) (2.0 g), 3-ethyl-5- (2-hydroxyethyl) -4 -While stirring a solution of methylthiazolium bromide (0.50 g, 2.0 mmol) and methyl acrylate (V-1) (0.36 ml, 4.0 mmol) in anhydrous THF (10 ml), DBU (0.30 ml , 2.0 mmol) in anhydrous THF (3 ml) was added. The reaction mixture was stirred at 50 ° C. for 10 hours, further compound (V-1) (0.18 ml, 2.0 mmol) was added, and the mixture was stirred at 50 ° C. for 10 hours. The reaction was diluted with ether and filtered through Celite-pad. The filtrate was washed successively with 1N hydrochloric acid, saturated multistory water, and water, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography to obtain 0.32 g (yield 48%) of the title compound (VI-1).

¹ H NMR (CDCl ₃ ) δ 1.14 (s, 9H), 1.45 (s, 9H), 2.57 (ddd, J = 17.2, 7.0, 7.0 Hz, 1H), 2.64 (ddd, J = 17.2, 7.0, 7.0 Hz , 1H), 2.84 (ddd, J = 18.3, 7.0, 7.0 Hz, 1H), 2.91 (ddd, J = 18.3, 7.0, 7.0 Hz, 1H), 3.55 (dd, J = 9.0, 4.2 Hz, 1H), 3.68 (s, 3H), 3.81 (dd, J = 9.0, 3.4 Hz, 1H), 4.31 (ddd, J = 7.6, 4.2, 3.4 Hz, 1H), 5.46 (d, J = 7.6 Hz, 1H).

(5) 5-Amino-6- (tert-butoxy) -4-oxohexanoic acid methyl trifluoroacetic acid (VII-1: R = Me, Y ³ = ^t Bu)
While stirring a solution of compound (VI-1) (0.24 g, 0.72 mmol) obtained in (4) above in methylene chloride (2 ml), trifluoroacetic acid (0.5 ml) was added at 0 ° C. to react. The solution was stirred at 0 ° C. for 2 hours. Subsequently, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 0.20 g (yield 80%) of the title compound (VII-1).

¹ H NMR (CDCl ₃ ) δ 1.17 (s, 9H), 2.65 (ddd, J = 17.6, 6.8, 6.8 Hz, 1H), 2.70 (ddd, J = 17.6, 6.8, 6.8 Hz, 1H), 2.82 (ddd , J = 18.3, 6.8, 6.8 Hz, 1H), 2.90 (ddd, J = 18.3, 6.8, 6.8 Hz, 1H), 3.68 (s, 3H), 3.90 (dd, J = 10.3, 3.4 Hz, 1H), 3.93 (dd, J = 10.3, 3.4 Hz, 1H), 4.42 (dd, J = 3.4, 3.4 Hz, 1H), 7.20-9.20 (br s, 3H).
¹⁹ F NMR (CDCl ₃ ) δ 85.81 (s, 3F).

(6) Methyl 6- (tert-butoxy) -4-oxo-5-[(3,3,3-trifluoro-1-oxopropyl) amino] hexanoate (Compound Ib-1)
While stirring a solution of 3,3,3-trifluoropropionic acid (46 μl, 0.52 mmol) in anhydrous methylene chloride (2 ml), oxalyl chloride (47 μl, 0.56 mmol) and DMF (3 drops) were added at 0 ° C. The reaction solution was stirred at room temperature for 3 hours. The reaction mixture was stirred into a solution of compound (VII-1) obtained in (5) above (0.14 g, 0.40 mmol) and triethylamine (0.28 ml, 2.0 mmol) in anhydrous methylene chloride (3 ml). The mixture was added at 0 ° C., and the reaction solution was stirred at room temperature all day and night. The reaction mixture was diluted with ethyl acetate, washed successively with water and brine, dried over anhydrous magnesium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography to obtain 78 mg (yield 58%) of the title compound (Ib-1).

¹ H NMR (CDCl ₃ ) δ 1.14 (s, 9H), 2.59 (ddd, J = 17.2, 6.8, 6.8 Hz, 1H), 2.68 (ddd, J = 17.2, 6.8, 6.8 Hz, 1H), 2.84 (ddd , J = 18.3, 6.8, 6.8 Hz, 1H), 2.89 (ddd, J = 18.3, 6.8, 6.8 Hz, 1H), 3.14 (q, J = 10.5 Hz, 2H), 3.63 (dd, J = 9.5, 3.9 Hz, 1H), 3.68 (s, 3H), 3.88 (dd, J = 9.5, 3.2 Hz, 1H), 4.69 (ddd, J = 6.8, 3.4, 3.2 Hz, 1H), 6.76 (d, J = 6.8 Hz , 1H).
¹⁹ F NMR (CDCl ₃ ) δ 98.73 (t, J = 10.5 Hz, 3F).
¹³ C NMR (CDCl ₃ ) δ 27.1, 27.6, 34.4, 41.6 (q, J = 29.9 Hz), 51.7, 59.2, 61.1, 73.6, 123.9 (q, J = 276.9 Hz), 162.2, 172.6, 204.7.

Methyl 6-hydroxy-4-oxo-5-[(3,3,3-trifluoro-1-oxopropyl) amino] hexanoate (Compound Ib-2)
While stirring a solution of compound (Ib-1) obtained in Example 3 (68 mg, 0.20 mmol) in methylene chloride (1.0 ml), trifluoroacetic acid (1.0 ml) was added at 0 ° C. Was stirred at room temperature for 3 hours. Then, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 36 mg (yield 63%) of the title compound (Ib-2).

¹ H NMR (CDCl ₃ ) δ 2.64-2.71 (m, 2H), 2.79-2.86 (m, 1H), 2.91-2.98 (m, 1H), 3.18 (q, J = 10.5 Hz, 2H), 3.25 (br s, 1H), 3.69 (s, 3H), 3.88 (dd, J = 12.0, 3.7 Hz, 1H), 4.22 (dd, J = 12.0, 2.9 Hz, 1H), 4.69 (ddd, J = 6.1, 3.7, 2.9 Hz, 1H), 7.13 (d, J = 6.1 Hz, 1H).
¹⁹ F NMR (CDCl ₃ ) δ 98.77 (t, J = 10.5 Hz, 3F).
¹³ C NMR (CDCl ₃ ) δ 27.7, 34.0, 41.3 (q, J = 29.3 Hz), 52.1, 61.0, 62.3, 123.8 (q, J = 276.9 Hz), 162.8, 173.8, 205.6.

Methyl 4-oxo-5-[(3,3,3-trifluoro-1-oxopropyl) amino] -5-hexenoate (Compound Ia-3)
While stirring a solution of compound (Ib-2) obtained in Example 4 (34 mg, 0.12 mmol) and DMAP (2.2 mg, 0.18 μmol) in anhydrous acetonitrile (1.0 ml), (Boc) ₂ O ( 28 μl, 0.12 mmol) was added and the reaction was stirred overnight at room temperature. The reaction mixture was diluted with ether, washed successively with 1M aqueous KHSO ₄ solution, saturated multilayered water, and brine, and dried over anhydrous magnesium sulfate. Then, the solvent was distilled off under reduced pressure to obtain 26 mg (yield 83%) of the title compound (Ia-3).

¹ H NMR (CDCl ₃ ) δ 2.69 (t, J = 6.5 Hz, 2H), 3.12 (t, J = 6.5 Hz, 2H), 3.19 (q, J = 10.5 Hz, 2H), 3.71 (s, 3H) , 5.94 (t, J = 1.4 Hz, 1H), 6.97 (d, J = 1.4 Hz, 1H), 8.27 (br s, 1H).
¹⁹ F NMR (CDCl ₃ ) δ 98.76 (t, J = 10.5 Hz, 3F).

6- (tert-Butoxy) -4-oxo-5-[(3,3,3-trifluoro-1-oxopropyl) amino] hexanoic acid tert-butyl ester (Compound Ib-3)

(1) 6- (tert-Butoxy) -5- (tert-butoxycarbonylamino) -4-oxohexanoic acid tert-butyl ester (VI-2: R = Y ³ = ^t Bu, Q = Boc)
Compound (IV-1) (4.9 g, 20 mmol) obtained in Example 3 (3), MS-4A (20 g), 3-ethyl-5- (2-hydroxyethyl) -4-methylthiazolium bromide While stirring a solution of acrylic acid tert-butyl ester (V-2) (5.8 ml, 40 mmol) in anhydrous THF (40 ml) and anhydrous THF (3.0 ml, 20 mmol) in anhydrous THF (3.0 g, 20 mmol) 10 ml) solution was added. The reaction mixture was stirred at 50 ° C. for 10 hours, further compound (V-2) (2.9 ml, 20 mmol) was added, and the mixture was stirred at 50 ° C. for 10 hours. The reaction was diluted with ether and filtered through Celite-pad. The filtrate was washed successively with 1N hydrochloric acid, saturated multistory water, and water, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography to obtain 3.0 g (yield 41%) of the title compound (VI-2).

¹ H NMR (CDCl ₃ ) δ 1.14 (s, 9H), 1.43 (s, 9H), 1.45 (s, 9H), 2.48 (ddd, J = 17.0, 7.1, 7.1 Hz, 1H), 2.54 (ddd, J = 17.0, 7.1, 7.1 Hz, 1H), 2.78 (ddd, J = 18.3, 7.1, 7.1 Hz, 1H), 2.84 (ddd, J = 18.3, 7.1, 7.1 Hz, 1H), 3.56 (dd, J = 9.2 , 4.1 Hz, 1H), 3.81 (dd, J = 9.2, 3.4 Hz, 1H), 4.31 (ddd, J = 7.6, 4.1, 3.4 Hz, 1H), 5.47 (d, J = 7.6 Hz, 1H).
¹³ C NMR (CDCl ₃ ) δ 27.2, 28.0, 28.2, 29.1, 35.0, 60.0, 61.8, 73.3, 79.6, 80.4, 155.4, 171.7, 206.8.

(2) 5-Amino-6- (tert-butoxy) -4-oxohexanoic acid tert-butyl ester / trifluoroacetic acid (VII-2: R = Y ³ = ^t Bu)
From the compound (VI-2) obtained in (1) above, the title compound (VII-2) was obtained in a yield of 67% according to the method described in Example 3 (5).

¹ H NMR (CDCl ₃ ) δ 1.17 (s, 9H), 1.43 (s, 9H), 2.57 (dd, J = 6.8, 6.6 Hz, 2H), 2.78 (dt, J = 18.3, 6.6 Hz, 1H), 2.84 (dt, J = 18.3, 6.8 Hz, 1H), 3.93 (d, J = 3.4 Hz, 2H), 4.38 (t, J = 3.4, 1H), 7.20-9.20 (br s, 3H).
¹⁹ F NMR (CDCl ₃ ) δ 86.02 (s, 3F).
¹³ C NMR (CDCl ₃ ) δ 26.9, 27.9, 28.9, 33.6, 58.9, 59.3, 74.5, 81.0, 171.3, 202.4.

(3) 6- (tert-Butoxy) -4-oxo-5-[(3,3,3-trifluoro-1-oxopropyl) amino] hexanoic acid tert-butyl ester (Compound Ib-3)
From the compound (VII-2) obtained in (2) above, the title compound (Ib-3) was obtained in a yield of 65% according to the method described in Example 3 (6).

¹ H NMR (CDCl ₃ ) δ 1.14 (s, 9H), 1.43 (s, 9H) 2.50 (dt, J = 17.2, 6.6 Hz, 1H), 2.59 (dt, J = 17.2, 6.8 Hz, 1H), 2.80 (dd, J = 6.8, 6.6 Hz, 2H), 3.13 (qd, J = 10.5, 1.1 Hz, 2H), 3.63 (dd, J = 9.5, 3.9 Hz, 1H), 3.88 (dd, J = 9.5, 3.2 Hz, 1H), 4.68 (ddd, J = 6.6, 3.9, 3.2 Hz, 1H), 6.76 (d, J = 6.6 Hz, 1H).
¹⁹ F NMR (CDCl ₃ ) δ 98.71 (t, J = 10.5 Hz, 3F).
¹³ C NMR (CDCl ₃ ) δ 27.1, 27.9, 29.1, 34.6, 41.6 (q, J = 29.9 Hz), 59.2, 61.0, 73.6, 80.7, 123.9 (q, J = 276.9 Hz), 162.1 (q, J = 3.2 Hz), 171.4, 204.8.

6-Hydroxy-4-oxo-5-[(3,3,3-trifluoro-1-oxopropyl) amino] hexanoic acid (Compound Ib-4)
Compound (Ib-3) obtained in Example 6 (48 mg, 0.13 mmol) was dissolved in trifluoroacetic acid (1 ml), and 3 drops of water were added. After the reaction solution was stirred at room temperature for 3 hours, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 31 mg (yield 91%) of the title compound (Ib-4).

¹ H NMR (DMSO-d ₆ ) δ 2.40 (dd, J = 6.8, 6.6 Hz, 2H), 2.72 (dt, J = 18.5, 6.6 Hz, 1H), 2.78 (dt, J = 18.5, 6.8 Hz, 1H ), 3.41 (qd, J = 11.3, 4.6 Hz, 2H), 3.60 (ddd, J = 11.2, 4.9, 4.6 Hz, 1H), 3.78 (ddd, J = 11.2, 5.4, 4.9 Hz, 1H), 4.41 ( ddd, J = 7.6, 4.9, 4.6 Hz, 1H), 5.05 (dd, J = 5.4, 4.9 Hz, 1H), 8.63 (d, J = 7.6 Hz, 1H), 12.15 (br s, 1H).
¹⁹ F NMR (DMSO-d ₆ ) δ 101.23 (t, J = 11.3 Hz, 3F).
¹³ C NMR (CD ₃ OD) δ 26.8, 34.0, 39.1 (q, J = 29.3 Hz), 60.3, 60.5, 123.9 (q, J = 275.8 Hz), 163.9 (q, J = 3.7 Hz), 174.7, 205.6 .

4-Oxo-5-[(3,3,3-trifluoro-1-oxopropyl) amino] -5-hexenoic acid (Compound Ia-4)
While stirring a solution of compound (Ib-4) obtained in Example 7 (31 mg, 0.11 mmol) and DMAP (1.3 mg, 11 μmol) in anhydrous acetonitrile (1.1 ml), (Boc) ₂ O (25 μl, 0.11 mmol) was added and the reaction was stirred at room temperature overnight. Subsequently, a solution of TMG (4.1 μl, 33 μmol) in anhydrous acetonitrile (0.1 ml) was added, and the reaction solution was reacted at room temperature for 2 days. The reaction mixture was diluted with ethyl acetate, washed with 0.3 M KHSO ₄ aqueous solution and brine, and dried over anhydrous magnesium sulfate. Subsequently, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 12 mg (43% yield) of the title compound (Ia-4).

¹ H NMR (CDCl ₃ ) δ 2.75 (t, J = 6.4 Hz, 2H), 3.12 (t, J = 6.4 Hz, 2H), 3.20 (q, J = 10.3 Hz, 2H), 5.94 (t, J = 1.3 Hz, 1H), 6.98 (d, J = 1.3 Hz, 1H), 8.27 (s, 1H).
¹⁹ F NMR (CDCl ₃ ) δ 98.74 (t, J = 10.3 Hz, 3F).
¹³ C NMR (CDCl ₃ ) δ 27.5, 30.3, 42.6 (q, J = 29.9 Hz), 110.9, 123.6 (q, J = 276.9 Hz), 137.2, 161.1 (q, J = 3.8 Hz), 176.5, 194.3.

6- (tert-Butoxy) -4-oxo-5-[(4,4,4-trifluoro-1-oxobutyl) amino] hexanoic acid tert-butyl ester (Compound Ib-5)
While stirring a solution of 4,4,4-trifluorobutyric acid (0.13 g, 0.94 mmol) in anhydrous methylene chloride (3 ml), oxalyl chloride (85 μl, 1.0 mmol) and DMF (3 drops) were added at 0 ° C. And the reaction was stirred at room temperature for 3 hours. The reaction mixture was added to a solution of compound (VII-2) obtained in Example 6 (2) (0.28 g, 0.72 mmol) and triethylamine (0.50 ml, 3.6 mmol) in anhydrous methylene chloride (10 ml). The mixture was added at 0 ° C. with stirring, and the reaction mixture was stirred at room temperature overnight. The reaction mixture was diluted with ethyl acetate, washed successively with water and brine, dried over anhydrous magnesium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography to obtain 0.18 g (yield 63%) of the title compound (Ib-5).

¹ H NMR (CDCl ₃ ) δ 1.14 (s, 9H), 1.44 (s, 9H) 2.45-2.54 (m, 5H), 2.58 (dt, J = 17.1, 7.1 Hz, 1H), 2.80 (ddd, J = 7.1, 7.1, 1.0 Hz, 2H), 3.59 (dd, J = 9.4, 4.1 Hz, 1H), 3.86 (dd, J = 9.4, 3.3 Hz, 1H), 4.66 (ddd, J = 6.8, 4.1, 3.3 Hz , 1H), 6.50 (d, J = 6.8 Hz, 1H).
¹⁹ F NMR (CDCl ₃ ) δ 94.94 (t, J = 10.0 Hz, 3F).
¹³ C NMR (CDCl ₃ ) δ 27.2, 28.0, 28.6 (q, J = 3.2 Hz), 29.1, 29.5 (q, J = 29.9 Hz), 34.8, 59.0, 61.3, 73.5, 80.6, 126.7 (q, J = 275.8 Hz), 169.3, 171.5, 205.4.

6-Hydroxy-4-oxo-5-[(4,4,4-trifluoro-1-oxobutyl) amino] hexanoic acid (Compound Ib-6)
The title compound (Ib-6) was obtained in 88% yield from the compound (Ib-5) obtained in Example 9 according to the method described in Example 7.

¹ H NMR (DMSO-d ₆ ) δ 2.38 (dd, J = 6.8, 6.6 Hz, 2H), 2.45-2.50 (m, 4H), 2.70 (dt, J = 18.7, 6.6 Hz, 1H), 2.76 (dt , J = 18.7, 6.8 Hz, 1H), 3.59 (dd, J = 10.7, 4.2 Hz, 1H), 3.72 (dd, J = 10.7, 5.0 Hz, 1H), 4.10 (br s, 1H), 4.36 (ddd , J = 7.3, 5.0, 4.2 Hz, 1H), 8.41 (d, J = 7.3 Hz, 1H), 12.14 (br s, 1H).
¹⁹ F NMR (DMSO-d ₆ ) δ 97.41 (t, J = 10.7 Hz, 3F).
¹³ C NMR (CD ₃ OD) δ 26.7, 27.1 (q, J = 3.2 Hz), 28.5 (q, J = 29.9 Hz), 34.0, 60.4, 126.5 (q, J = 275.3 Hz), 171.0, 174.4, 206.1 .

4-Oxo-5-[(4,4,4-trifluoro-1-oxobutyl) amino] -5-hexenoic acid (Compound Ia-5)
While stirring a solution of compound (Ib-6) obtained in Example 10 (61 mg, 0.21 mmol) and DMAP (2.6 mg, 21 μmol) in anhydrous acetonitrile (2.1 ml), (Boc) ₂ O (48 μl, 0.21 mmol) was added and the reaction was stirred at room temperature overnight. Subsequently, a solution of TMG (7.9 μl, 63 μmol) in anhydrous acetonitrile (0.2 ml) was added, and the reaction solution was reacted at room temperature for 24 hours. The reaction mixture was diluted with ethyl acetate, washed with 0.3 M KHSO ₄ aqueous solution and brine, and dried over anhydrous magnesium sulfate. Next, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 29 mg (yield 52%) of the title compound (Ia-5).

¹ H NMR (CDCl ₃ ) δ 2.47-2.62 (m, 4H), 2.73 (t, J = 6.4 Hz, 2H), 3.11 (t, J = 6.4 Hz, 2H), 5.88 (t, J = 1.0 Hz, 1H), 6.93 (d, J = 1.0 Hz, 1H), 8.09 (s, 1H).
¹⁹ F NMR (CDCl ₃ ) δ 94.98 (t, J = 10.0 Hz, 3F).
¹³ C NMR (CDCl ₃ ) δ 27.7, 29.1 (q, J = 29.9 Hz), 29.7 (q, J = 3.2 Hz), 30.3, 110.0, 126.5 (q, J = 275.8 Hz), 137.5, 168.5, 177.3, 194.6.

4-Oxo-5-[(3,3,3-trifluoro-1-oxopropyl) amino] pentanoic acid tert-butyl ester (Compound Ic-1)

(1) 2- (tert-Butoxycarbonylamino) ethanal (XII-1: Q = Boc)
The title compound (XII-1) was obtained in a yield of 82% according to the method described in Example 3 (3) using N-tert-butoxycarbonylaminoglycine methyl ester (XI-1: Q = Boc). .

¹ H NMR (CDCl ₃ ) δ 1.46 (s, 9H), 4.08 (d, J = 4.9 Hz, 2H), 5.20 (br s 1H), 9.66 (s, 1H).
¹³ C NMR (CDCl ₃ ) δ 28.2, 51.3, 80.1, 155.5, 196.9.

(2) 5- (tert-Butoxycarbonylamino) -4-oxopentanoic acid tert-butyl ester (Compound XIII-1: R = ^t Bu; Q = Boc)
Compound (XII-1) obtained in the above (1) (0.42 g, 2.6 mmol), MS-4A (2.6 g), 3-ethyl-5- (2-hydroxyethyl) -4-methylthiazo While stirring a solution of lithium bromide (0.20 g, 0.78 mmol) and acrylic acid tert-butyl ester (V-2) (0.76 ml, 5.2 mmol) in anhydrous THF (8 ml), DBU (0.24 ml, 1.6 mmol) in anhydrous THF (3 ml) was added. After stirring the reaction mixture for 10 hours while refluxing, Compound (V-2) (0.38 ml, 2.6 mmol) was further added and stirring was continued for 10 hours while refluxing. The reaction was diluted with ether and filtered through Celite-pad. The filtrate was washed successively with 1N hydrochloric acid, saturated multistory water, and water, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography to obtain 0.26 g (yield 35%) of the title compound (XIII-1).

¹ H NMR (CDCl ₃ ) δ 1.43 (s, 9H), 1.44 (s, 9H), 2.57 (t, J = 6.4 Hz, 2H), 2.67 (t, J = 6.4 Hz, 2H), 4.07 (d, J = 4.9, 2H), 5.22 (br s, 1H).
¹³ C NMR (CDCl ₃ ) δ 27.9, 28.2, 29.0, 34.4, 50.2, 79.7, 80.8, 155.5, 171.4, 204.2.

(3) 5-Amino-4-oxopentanoic acid tert-butyl ester / trifluoroacetate (Compound XIV-1: R = ^t Bu)
To a solution of compound (XIII-1) obtained in (2) (0.15 g, 0.52 mmol) in methylene chloride (2 ml) was added trifluoroacetic acid (0.22 ml) at 0 ° C., and 2 at 0 ° C. Stir for hours. Then, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 0.10 g (yield 63%) of the title compound (XIV-1).

¹ H NMR (CD ₃ OD) δ 1.43 (s, 9H), 2.58 (t, J = 6.3 Hz, 2H), 2.75 (t, J = 6.3 Hz, 2H), 4.01 (s, 2H).
¹⁹ F NMR (CD ₃ OD) δ 88.38 (s, 3F).
¹³ C NMR (CD ₃ OD) δ 26.4, 28.0, 33.5, 46.3, 80.2, 171.5, 201.2.

(4) 4-Oxo-5-[(3,3,3-trifluoro-1-oxopropyl) amino] pentanoic acid tert-butyl ester (Compound Ic-1)
From the compound (XIV-1) obtained in (3) above, the title compound (Ic-1) was obtained in a yield of 74% according to the method described in Example 3 (6).

¹ H NMR (CDCl ₃ ) δ 1.43 (s, 9H) 2.60 (t, J = 6.3 Hz, 2H), 2.69 (t, J = 6.3 Hz, 2H), 3.14 (q, J = 10.5 Hz, 2H), 4.26 (d, J = 4.4 Hz, 2H), 6.64 (br s, 1H).
¹⁹ F NMR (CDCl ₃ ) δ 98.69 (t, J = 10.5 Hz, 3F).
¹³ C NMR (CDCl ₃ ) δ 27.9, 29.0, 34.7, 41.3 (q, J = 29.9 Hz), 49.3, 81.0, 123.8 (q, J = 276.9 Hz), 162.4 (q, J = 3.7 Hz), 171.3, 203.2.

4-Oxo-5-[(3,3,3-trifluoro-1-oxopropyl) amino] pentanoic acid (Compound Ic-2)
The title compound (Ic-2) was obtained in 88% yield from the compound (Ic-1) obtained in Example 12 according to the method described in Example 7.

¹ H NMR (DMSO-d ₆ ) δ 2.43 (t, J = 6.4 Hz, 2H), 2.65 (t, J = 6.4 Hz, 2H), 3.34 (q, J = 10.7 Hz, 2H), 4.05 (d, J = 5.4 Hz, 2H), 8.55 (t, J = 5.4 Hz, 1H), 12.19 (br s, 1H).
¹⁹ F NMR (DMSO-d ₆ ) δ 101.09 (t, J = 10.7 Hz, 3F).
¹³ C NMR (CD ₃ OD) δ 26.7, 33.4, 39.2 (q, J = 29.4 Hz), 48.0, 123.9 (q, J = 276.4 Hz), 164.0 (q, J = 3.7 Hz), 174.2, 203.8.

(PBGS inhibitory activity)
The activity of PBG synthase (PBGS) can be determined by converting PBG produced by PBGS into a compound that develops color by referring to the method of Michel et al. (Genetics and Molecular Research, 2003, 2, 48-62). Was quantified and evaluated as the activity of PBGS.

  4 μg of PBGS (HemB protein) and 5 mM chloride in 0.5 ml of 50 mM 2- [4- (2-hydroxyethyl) -1-piperazinyl-ethanesulfonic acid sodium salt (HEPES-Na) buffer (pH 8.0) Magnesium was added in advance and allowed to stand at room temperature for 10 minutes. Next, 2.5 μmol of 5-ALA as a substrate was added and reacted at 37 ° C. for 15 minutes. After the reaction, 50 μl of 50% trichloroacetic acid solution was added to stop the reaction, and the mixture was allowed to stand on ice for 15 minutes. Centrifugation was performed at 4 ° C. at 15,000 rpm × 10 min, 0.5 ml of the supernatant was collected, and 0.5 ml of Erich reagent (2% p-dimethylaminobenzaldehyde (DMAB) solution adjusted with 6N HCl) was added. After 15 minutes at room temperature, color development was measured by absorbance at 555 nm.

The test compound IC ₅₀ is obtained by adding a test compound adjusted with 50 mM HEPES-Na buffer (pH 8.0) or DMSO to the reaction solution before adding the substrate, measuring the activity of PBGS, and adding the test compound. The concentration at which 50% inhibition was achieved was calculated from the inhibition curve, with no condition being 100% activity.
The results are shown in Table 4.

[Formulation Example 1]
A tablet is prepared by a conventional method using a composition comprising 10 mg of compound (Ia-2), 70 mg of lactose, 15 mg of starch, 4 mg of polyvinyl alcohol and 1 mg of magnesium stearate (total 100 mg).

[Formulation Example 2]
In a conventional manner, 70 ml of compound (Ia-5), 50 mg of purified soybean oil, 10 mg of egg yolk lecithin and 25 mg of glycerin are added with distilled water for injection to a total volume of 100 mL, filled in a vial, and then heat-sterilized. To prepare an injection.

The novel araremycin derivative of the present invention has excellent PBGS inhibitory activity and can be used as a therapeutic agent for infectious diseases. In particular, and it is known to have antibacterial activity arare mycin is 5-ALA as an inhibitor of PBGS to synthesize PBG as a substrate, the PBGS multidrug resistance aeruginosa ^{Mg 2+} type of opportunistic infections in hospitals Therefore, the compound (I) of the present invention in which antibacterial activity is enhanced by modifying the structure of aralemycin which is effective in inhibiting Mg ²⁺ type PBGS is expected to be used as a highly effective antibacterial agent.

Claims (8)

Formula (I)

(In the formula, R represents a hydrogen atom or a lower alkyl group, X represents a halogen-substituted methyl group, Y ¹ and Y ² represent a hydrogen atom when one represents a hydrogen atom, and the other represents a hydrogen atom or a hydroxymethyl group. Or a lower alkoxymethyl group, or Y ¹ and Y ² together represent a double bond, and m represents an integer of 0 to 5).
An araremycin derivative represented by the formula or a pharmacologically acceptable salt thereof. The compound represented by the formula (I) is represented by the following formula (Ia)

(Wherein R, X and m are as defined above)
The araremycin derivative or a pharmacologically acceptable salt thereof according to claim 1, which is represented by the formula: R is a hydrogen atom, The araremycin derivative or its pharmacologically acceptable salt of Claim 1 or 2 characterized by the above-mentioned. m is an integer of 0-2, The araremycin derivative in any one of Claims 1-3, or its pharmacologically acceptable salt. X is a fluorine-substituted methyl group, The aralemycin derivative or pharmacologically acceptable salt thereof according to any one of claims 1 to 4. 6. The aralemycin derivative or a pharmacologically acceptable salt thereof according to claim 5, wherein the fluorine-substituted methyl group is a trifluoromethyl group. An inhibitor of porphobilinogen synthase (PBGS) comprising the araremycin derivative according to any one of claims 1 to 6 or a pharmacologically acceptable salt thereof as an active ingredient. The antibacterial agent which contains the araremycin derivative in any one of Claims 1-6, or its pharmacologically acceptable salt as an active ingredient.