JPWO2010041699A1 - BTMA2 compound - Google Patents

Abstract

A compound represented by the following general formula (1) or a salt thereof. [Wherein, X represents a group represented by any one selected from the group consisting of the following general formulas (2) to (5). In the general formulas (2) to (5), R21 represents a hydrogen atom, a hydroxy group, an alkyl group, an alkylthio group or an amino group, R31, R32, R41 and R42 each represents a hydrogen atom or an alkyl group, and R51, R52 And R53 each represents a hydrogen atom, a hydroxy group, an alkyl group, a mercapto group, an alkylthio group, an amino group, an alkoxy group, a phosphoric acid group or an aromatic heterocyclic group. ]

Description

The present invention relates to BTMA ₂ compounds.

Conventionally used antibiotics include β-lactams, aminoglycosides, macrolides, and tetracyclines, which play a major role in modern infection treatment. However, heavy use of antibiotics has resulted in the emergence of resistant bacteria, which has become a serious problem in recent years. MRSA (methicillin-resistant Staphylococcus aureus (methicillin-resistant Staphylococcus aureus )) is mentioned as one of resistant bacteria.

Bottromycin A ₂ (BTMA ₂ ) is known as one of therapeutic agents for MRSA (see, for example, Patent Document 1). However, BTMA ₂ is potent and exhibits a broad antibacterial spectrum in vitro, but is known to lose its antibacterial activity in vivo with oral or subcutaneous administration.

International Publication No. 2006/103010 Pamphlet

An object of the present invention is to provide a BTMA ₂ compound exhibiting excellent antibacterial activity and a broad antibacterial spectrum. Another object of the present invention is to provide a BTMA ₂ compound that does not lose its antibacterial activity even when administered orally or subcutaneously. Furthermore, an object of the present invention is to provide an antibacterial agent which exhibits excellent antibacterial activity and a broad antibacterial spectrum, and does not lose its antibacterial activity even when administered orally or subcutaneously.

As a result of intensive studies on the above problems, the present inventors have found that a BTMA ₂ compound having a structure that has not been known so far exhibits excellent antibacterial activity and a broad antibacterial spectrum. Further, a BTMA ₂ compound was found that does not lose its antibacterial activity even when administered orally or subcutaneously. The present invention has been completed based on these findings.

The present invention provides the following means.
[1] A compound represented by the following general formula (1) or a salt thereof.

[Wherein, X represents a group represented by any one selected from the group consisting of the following general formulas (2) to (5).

In the general formula (2), R ²¹ represents a hydrogen atom, a hydroxy group, an alkyl group, an alkylthio group or an amino group.

In general formula (3), R ³¹ and R ³² each independently represent a hydrogen atom or an alkyl group, and may be bonded to each other to form a ring.

In the general formula (4), R ⁴¹ and R ⁴² each independently represent a hydrogen atom or an alkyl group.

In general formula (5), R ⁵¹ , R ⁵² and R ⁵³ are each independently a hydrogen atom, a hydroxy group, an alkyl group, a mercapto group, an alkylthio group, an amino group, an alkoxy group, a phosphate group or an aromatic heterocyclic group. And may be bonded to each other to form a ring. ]
[2] The group represented by the general formula (2) is represented by any one selected from the group consisting of the general formula (2-1), the general formula (2-2), and the general formula (2-3). The compound or a salt thereof according to the above item [1], wherein the compound or a salt thereof.

[In General Formula (2-1), R ¹²¹ and R ¹²² each independently represent a hydrogen atom or an alkyl group, and may be bonded to each other to form a ring.

In General Formula (2-2), R ²²¹ represents a hydrogen atom, a hydroxy group, or an alkyl group.

In general formula (2-3), ^R321 represents an alkyl group. ]
[3] The group represented by general formula (5) is selected from the group consisting of general formula (5-1), general formula (5-2), general formula (5-3), and general formula (5-4). The compound or a salt thereof according to the above item [1], which is a group represented by any one of

[In General Formula (5-1), R ¹⁵¹ and R ¹⁵² each independently represent an alkyl group, and may be bonded to each other to form a ring. R ¹⁵³ represents a hydrogen atom or an alkyl group.

In general formula (5-2), ^R252 represents a hydrogen atom, an alkyl group, or an aromatic heterocyclic group.

In general formula (5-3), R ³⁵¹ and R ³⁵² each independently represent a hydrogen atom or an alkyl group, and may be bonded to each other to form a ring.

In general formula (5-4), ^R451 and ^R452 each independently represents an alkyl group, and may be bonded to each other to form a ring. ]
[4] An antibacterial agent containing at least one of the compound or salt thereof according to any one of [1] to [3].
[5] Use of the compound or salt thereof according to any one of [1] to [3] as an antibacterial agent.
[6] The compound according to any one of [1] to [3] or a salt thereof for use in the treatment of an infection caused by MRSA.
[7] A method for treating infection by MRSA, comprising administering the compound or salt thereof according to any one of [1] to [3].

The BTMA ₂ compounds of the present invention exhibit excellent antimicrobial activity and a broad antimicrobial spectrum. In addition, the BTMA ₂ compound of the present invention does not lose its antibacterial activity even when administered orally or subcutaneously. Furthermore, the antibacterial agent of the present invention exhibits excellent antibacterial activity and a broad antibacterial spectrum, and the antibacterial activity does not disappear even when administered orally or subcutaneously.

  These and other features and advantages of the present invention will become more apparent from the following description.

Hereinafter, the present invention will be described in detail based on preferred embodiments thereof.
In the general formula (1), X represents a group represented by any one of the general formulas (2) to (5). The compounds of the present invention exhibit excellent antimicrobial activity and a broad antimicrobial spectrum.

  First, general formula (2) will be described.

In the general formula (2), R ²¹ represents a hydrogen atom, a hydroxy group, an alkyl group, an alkylthio group or an amino group. As the alkyl group represented by R ²¹ , an alkyl group having 1 to 12 carbon atoms is preferable, an alkyl group having 1 to 10 carbon atoms is more preferable, and an alkyl group having 1 to 8 carbon atoms is more preferable. As the alkylthio group represented by R ²¹ , an alkylthio group having 1 to 8 carbon atoms is preferable, an alkylthio group having 1 to 4 carbon atoms is preferable, and an alkylthio group having 1 to 3 carbon atoms is more preferable. The alkyl group, alkylthio group and amino group represented by R ²¹ may further have a substituent.

Examples of the substituent include halogen atoms (for example, fluorine atom, chlorine atom, bromine atom, iodine atom), alkyl groups (for example, methyl group, ethyl group, n-propyl group, isopropyl group, tert-butyl group, n -Octyl group, 2-ethylhexyl group), cycloalkyl group (for example, cyclohexyl group, cyclopentyl group, 4-n-dodecylcyclohexyl group), bicycloalkyl group (for example, bicyclo [1,2,2] heptan-2-yl Group, bicyclo [2,2,2] octane-3-yl group), alkenyl group (for example, vinyl group, allyl group), cycloalkenyl group (for example, 2-cyclopenten-1-yl, 2-cyclohexene-1- Yl group), bicycloalkenyl group (for example, bicyclo [2,2,1] hept-2-en-1-yl group, bicyclo [2 2,2] oct-2-en-4-yl group), alkynyl group (for example, ethynyl group, propargyl group), aryl group (for example, phenyl group, p-tolyl group, naphthyl group), heterocyclic group (for example, 2-furyl group, 2-thienyl group, 2-pyrimidinyl group, 2-benzothiazolyl group), cyano group, hydroxyl group, nitro group, carboxyl group, alkoxy group (for example, methoxy group, ethoxy group, isopropoxy group, tert- Butoxy group, n-octyloxy group, 2-methoxyethoxy group), aryloxy group (for example, phenoxy group, 2-methylphenoxy group, 4-tert-butylphenoxy group, 3-nitrophenoxy group, 2-tetradecanoyl group) Aminophenoxy group), silyloxy group (for example, trimethylsilyloxy group, tert-butyldi) Tilsilyloxy group), heterocyclic oxy group (for example, 1-phenyltetrazol-5-oxy group, 2-tetrahydropyranyloxy group), acyloxy group (for example, formyloxy group, acetyloxy group, pivaloyloxy group, stearoyloxy) Group, benzoyloxy group, p-methoxyphenylcarbonyloxy group), carbamoyloxy group (for example, N, N-dimethylcarbamoyloxy group, N, N-diethylcarbamoyloxy group, morpholinocarbonyloxy group, N, N-di-) n-octylaminocarbonyloxy group, Nn-octylcarbamoyloxy group), alkoxycarbonyloxy group (for example, methoxycarbonyloxy group, ethoxycarbonyloxy group, tert-butoxycarbonyloxy group, n-octylcarbonyl group) Ruoxy group), aryloxycarbonyloxy group (for example, phenoxycarbonyloxy group, p-methoxyphenoxycarbonyloxy group, pn-hexadecyloxyphenoxycarbonyloxy group), amino group (for example, amino group, methylamino group, Dimethylamino group, anilino group, N-methyl-anilino group, diphenylamino group), acylamino group (for example, formylamino group, acetylamino group, pivaloylamino group, lauroylamino group, benzoylamino group), aminocarbonylamino group (for example, , Carbamoylamino group, N, N-dimethylaminocarbonylamino group, N, N-diethylaminocarbonylamino group, morpholinocarbonylamino group), alkoxycarbonylamino group (for example, methoxycarbonylamino group, Toxylcarbonylamino group, tert-butoxycarbonylamino group, n-octadecyloxycarbonylamino group, N-methyl-methoxycarbonylamino group), aryloxycarbonylamino group (for example, phenoxycarbonylamino group, p-chlorophenoxycarbonylamino group) , Mn-octyloxyphenoxycarbonylamino group), sulfamoylamino group (for example, sulfamoylamino group, N, N-dimethylaminosulfonylamino group, Nn-octylaminosulfonylamino group), alkyl or Arylsulfonylamino group (for example, methylsulfonylamino group, butylsulfonylamino group, phenylsulfonylamino group, 2,3,5-trichlorophenylsulfonylamino group, p-methylphenylsulfonylamino group) Group), mercapto group, alkylthio group (for example, methylthio group, ethylthio group, n-hexadecylthio group), arylthio group (for example, phenylthio group, p-chlorophenylthio group, m-methoxyphenylthio group), heterocyclic thio group (For example, 2-benzothiazolylthio group, 1-phenyltetrazol-5-ylthio group), sulfamoyl group (for example, N-ethylsulfamoyl group, N- (3-dodecyloxypropyl) sulfamoyl group, N, N -Dimethylsulfamoyl group, N-acetylsulfamoyl group, N-benzoylsulfamoyl group, N- (N'-phenylcarbamoyl) sulfamoyl group), sulfo group, alkyl or arylsulfinyl group (for example, methylsulfinyl group) , Ethylsulfinyl group, phenylsulfinyl group , P-methylphenylsulfinyl group), alkyl or arylsulfonyl group (for example, methylsulfonyl group, ethylsulfonyl group, phenylsulfonyl group, p-methylphenylsulfonyl group), acyl group (for example, acetyl group, pivaloylbenzoyl group) ), Aryloxycarbonyl group (for example, phenoxycarbonyl group, o-chlorophenoxycarbonyl group, m-nitrophenoxycarbonyl group, p-tert-butylphenoxycarbonyl group), alkoxycarbonyl group (for example, methoxycarbonyl group, ethoxycarbonyl group) , Tert-butoxycarbonyl group, n-octadecyloxycarbonyl group), carbamoyl group (for example, carbamoyl group, N-methylcarbamoyl group, N, N-dimethylcarbamoyl group, N, N-di-n-) Octylcarbamoyl group, N- (methylsulfonyl) carbamoyl group), aryl or heterocyclic azo group (for example, phenylazo group, p-chlorophenylazo group, 5-ethylthio-1,3,4-thiadiazol-2-ylazo group), Imido group (for example, N-succinimide group, N-phthalimide group), phosphino group (for example, dimethylphosphino group, diphenylphosphino group, methylphenoxyphosphino group), phosphinyl group (for example, phosphinyl group, dioctyloxyphosphini) Group, diethoxyphosphinyl group), phosphinyloxy group (for example, diphenoxyphosphinyloxy group, dioctyloxyphosphinyloxy group), phosphinylamino group (for example, dimethoxyphosphinylamino group) , Dimethylaminophosphinylamino ), Silyl groups (e.g., trimethylsilyl group, tert- butyldimethylsilyl group, and the like phenyl dimethylsilyl group).
Further, the substituent may be further substituted, and when there are a plurality of substituents, they may be the same or different. At that time, examples of the substituent include the above-described monovalent substituent. Moreover, you may combine with substituents and may form a ring.

In the general formula (2), R ²¹ is preferably a hydrogen atom, an alkyl group, an alkylthio group or an amino group, and more preferably an alkyl group or an amino group.

  Moreover, it is preferable that group represented by General formula (2) is group represented by General formula (2-1), General formula (2-2), or General formula (2-3). Hereinafter, general formula (2-1), general formula (2-2), and general formula (2-3) will be described.

In general formula (2-1), R ¹²¹ and R ¹²² each independently represent a hydrogen atom or an alkyl group, and may be bonded to each other to form a ring. As the alkyl group represented by R ¹²¹ and R ¹²² , an alkyl group having 1 to 6 carbon atoms is preferable, an alkyl group having 1 to 4 carbon atoms is more preferable, and an alkyl group having 1 to 2 carbon atoms is more preferable. The alkyl group represented by R ¹²¹ and R ¹²² may further have a substituent. Examples of the substituent include a substituent that R ²¹ may have.
In General Formula (2-1), R ¹²¹ and R ¹²² are each independently preferably a hydrogen atom, or a substituted or unsubstituted methyl group or ethyl group.

In General Formula (2-2), R ²²¹ represents a hydrogen atom, a hydroxy group, or an alkyl group. The alkyl group represented by R ²²¹ is preferably an alkyl group having 1 to 12 carbon atoms, more preferably an alkyl group having 1 to 8 carbon atoms, and more preferably an alkyl group having 2 to 4 carbon atoms. The alkyl group represented by R ²²¹ may further have a substituent. Examples of the substituent include a substituent that R ²¹ may have.
In General Formula (2-2), R ²²¹ is preferably an alkyl group, and more preferably a substituted or unsubstituted ethyl group, propyl group, or butyl group.

In general formula (2-3), ^R321 represents an alkyl group. As the alkyl group represented by ^R321 , an alkyl group having 1 to 4 carbon atoms is preferable, an alkyl group having 1 to 3 carbon atoms is preferable, and an alkyl group having 2 to 3 carbon atoms is more preferable. The alkyl group represented by R ³²¹ may further have a substituent. Examples of the substituent include a substituent that R ²¹ may have.
In General Formula (2-3), R ³²¹ is more preferably a methyl group, a propyl group, or an isopropyl group.

  Next, general formula (3) will be described.

In general formula (3), R ³¹ and R ³² each independently represent a hydrogen atom or an alkyl group, and may be bonded to each other to form a ring. As the alkyl group represented by R ³¹ and R ³² , an alkyl group having 1 to 4 carbon atoms is preferable, and an alkyl group having 1 to 3 carbon atoms is more preferable. The alkyl group represented by R ³¹ and R ³² may further have a substituent. Examples of the substituent include a substituent that R ²¹ may have.
In the general formula (3), R ³¹ and R ³² are preferably each independently a hydrogen atom, or a substituted or unsubstituted methyl group, ethyl group, or isopropyl group.

  Next, general formula (4) will be described.

In the general formula (4), R ⁴¹ and R ⁴² each independently represent a hydrogen atom or an alkyl group. As the alkyl group represented by R ⁴¹ and R ⁴² , an alkyl group having 1 to 6 carbon atoms is preferable, an alkyl group having 1 to 4 carbon atoms is more preferable, and an alkyl group having 1 to 3 carbon atoms is more preferable. The alkyl group represented by R ⁴¹ and R ⁴² may further have a substituent. Examples of the substituent include a substituent that R ²¹ may have.
In the general formula (4), R ⁴¹ is preferably a hydrogen atom or an alkyl group having 1 to 3 carbon atoms, more preferably an alkyl group having 1 to 3 carbon atoms, and further preferably a propyl group. R ⁴² is preferably an alkyl group having 1 to 3 hydrogen atoms or carbon atoms, more preferably an alkyl group having 1 to 3 carbon atoms, more preferably methyl group.

  Next, general formula (5) will be described.

In general formula (5), R ⁵¹ , R ⁵² and R ⁵³ are each independently a hydrogen atom, a hydroxy group, an alkyl group, a mercapto group, an alkylthio group, an amino group, an alkoxy group, a phosphate group or an aromatic heterocyclic group. And may be bonded to each other to form a ring. As the alkyl group represented by R ⁵¹ , R ⁵² and R ⁵³ , an alkyl group having 1 to 4 carbon atoms is preferable, and an alkyl group having 2 to 3 carbon atoms is more preferable. As the alkylthio group represented by R ⁵¹ , R ⁵² and R ⁵³ , an alkylthio group having 1 to 4 carbon atoms is preferable, and an alkylthio group having 1 to 3 carbon atoms is more preferable. As the amino group represented by R ⁵¹ , R ⁵² and R ⁵³ , an amino group having 1 to 3 carbon atoms is preferable, and an amino group having 1 to 2 carbon atoms is more preferable. As the alkoxy group represented by R ⁵¹ , R ⁵² and R ⁵³ , an alkoxy group having 1 to 3 carbon atoms is preferable. The aromatic heterocyclic group represented by R ⁵¹ and R ⁵² is preferably a 5- or 6-membered aromatic heterocyclic group, more preferably a 5-membered aromatic heterocyclic group. The alkyl group, alkylthio group, amino group, alkoxy group, phosphoric acid group and aromatic heterocyclic group represented by R ⁵¹ , R ⁵² and R ⁵³ may further have a substituent. Examples of the substituent include a substituent that R ²¹ may have.

  In addition, the group represented by the general formula (5) is a group represented by the general formula (5-1), the general formula (5-2), the general formula (5-3), or the general formula (5-4). It is preferable that Hereinafter, general formula (5-1), general formula (5-2), general formula (5-3), and general formula (5-4) will be described.

In general formula (5-1), R ¹⁵¹ and R ¹⁵² each independently represent an alkyl group, and may be bonded to each other to form a ring. R ¹⁵³ represents a hydrogen atom or an alkyl group. As the alkyl group represented by R ¹⁵¹ , R ¹⁵² and R ¹⁵³ , an alkyl group having 1 to 4 carbon atoms is preferable, and an alkyl group having 1 to 3 carbon atoms is more preferable. The alkyl group represented by R ¹⁵¹ , R ¹⁵² and R ¹⁵³ may further have a substituent. Examples of the substituent include a substituent that R ²¹ may have.
In General Formula (5-1), R ¹⁵¹ and R ¹⁵² are each independently preferably a methyl group, an ethyl group, or a propyl group, and more preferably a methyl group or an ethyl group. R ¹⁵³ is preferably a hydrogen atom, a methyl group, an ethyl group or a propyl group, more preferably a hydrogen atom.

In general formula (5-2), ^R252 represents a hydrogen atom, an alkyl group, or an aromatic heterocyclic group. As the alkyl group represented by ^R252 , an alkyl group having 1 to 4 carbon atoms is preferable, and an alkyl group having 1 to 3 carbon atoms is more preferable. As the aromatic heterocycle represented by R ²⁵² , a 5- to 6-membered aromatic heterocyclic group is preferable, and a 5- to 6-membered aromatic containing an oxygen atom, a sulfur atom and / or a nitrogen atom as a hetero atom. More preferred are group heterocyclic groups. The alkyl group represented by R ²⁵² and the aromatic heterocyclic ring may further have a substituent. Examples of the substituent include a substituent that R ²¹ may have.
In General Formula (5-2), R ²⁵² is preferably a hydrogen atom, a methyl group, an ethyl group, a propyl group, or a 2-thiazolyl group, and more preferably a hydrogen atom, an ethyl group, a propyl group, or a 2-thiazolyl group.

In general formula (5-3), R ³⁵¹ and R ³⁵² each independently represent a hydrogen atom or an alkyl group, and may be bonded to each other to form a ring. The alkyl group represented by R ³⁵¹ and R ³⁵² is preferably an alkyl group having 1 to 4 carbon atoms, and more preferably an alkyl group having 1 to 2 carbon atoms. The alkyl group represented by R ³⁵¹ and R ³⁵² may further have a substituent. Examples of the substituent include a substituent that R ²¹ may have.
In General Formula (5-3), R ³⁵¹ and R ³⁵² are each independently preferably a hydrogen atom, a methyl group, an ethyl group, or a propyl group, and more preferably a hydrogen atom, a methyl group, or an ethyl group.

In general formula (5-4), ^R451 and ^R452 each independently represents an alkyl group, and may be bonded to each other to form a ring. As the alkyl group represented by R ⁴⁵¹ and R ⁴⁵² , an alkyl group having 1 to 4 carbon atoms is preferable, and an alkyl group having 1 to 2 carbon atoms is more preferable. The alkyl group represented by R ⁴⁵¹ and R ⁴⁵² may further have a substituent. Examples of the substituent include a substituent that R ²¹ may have.
In General Formula (5-4), R ⁴⁵¹ and R ⁴⁵² are each independently preferably a methyl group, an ethyl group, or a propyl group, and more preferably a methyl group or an ethyl group.

  Specific examples of the compound represented by the general formula (1) are shown below, but the present invention is not limited thereto. In this specification, Ac, Me, Et, and Bn represent an acetyl group, a methyl group, an ethyl group, and a benzyl group, respectively. Moreover, the wavy line in the group represented by X represents the bonding site to the carbon atom to which X in the general formula (1) is bonded.

  The compound represented by the general formula (1) can be synthesized by any method. For example, the compound represented by the general formula (1) can be synthesized with reference to the following scheme. However, the present invention is not limited to this.

(Synthesis Example 1)
Hydrazine is added to an ethanol solution of BTMA ₂ and refluxed for 4 hours to prepare a hydrazide form of BTMA ₂ . Subsequently, the hydrazide is dissolved in N-methyl-2-pyrrolidinone (NMP) at 0 ° C., 4N hydrochloric acid / dioxane solution and n-butylnitrile are added, and the mixture is stirred for 30 minutes to prepare an intermediate (acyl azide). To do. Next, triethylamine is added to make the reaction solution basic, and then various amines are added and heated to reflux at 60 ° C. to obtain an amine body (BTM amide).

(Synthesis Example 2)
Triethylamine is added to the intermediate (acyl azide) prepared in the same manner as in Synthesis Example 1 to make the reaction solution basic, and then various thiols are added and stirred at room temperature to obtain a thioester (BTM thioester). .

(Synthesis Example 3)
Triethylamine is added to the intermediate (acyl azide) prepared in the same manner as in Synthesis Example 1 to make the reaction solution basic. Next, the mixture is extracted with ethyl acetate by a liquid separation operation to prepare a mixture of isocyanate and azido ester. The toluene solution of this mixture was stirred at 60 ° C. for 2 hours, and it was confirmed by LC / MS analysis that the acyl azide had disappeared and was all converted to an isocyanate form. Then, various amines were added to the reaction solution, and 60 ° C. The urea body (BTM urea) can be obtained by stirring for 4 hours.

(Synthesis Example 4)
A hydrazide of BTMA ₂ prepared in the same manner as in Synthesis Example 1 is dissolved in N, N-dimethylformamide / tetrahydrofuran at 0 ° C., a 4N hydrochloric acid / dioxane solution and n-butylnitrile are added, and the mixture is stirred for 30 minutes. Next, triethylamine is added to make the reaction mixture basic, t-butoxycarbonylpiperazine is added, and the mixture is heated to reflux at 40 ° C. to obtain Exemplary Compound (2-16). Next, 4N hydrochloric acid / dioxane is added to a dichloromethane solution of the exemplified compound (2-16), and the mixture is stirred at 0 ° C. for 2 hours to remove the t-butoxycarbonyl group, thereby obtaining the exemplified compound (2-17). Furthermore, the exemplary compound (2-17) is dissolved in N, N-dimethylformamide, various alkyl halides and K ₂ CO ₃ are added, and the mixture is stirred at 40 ° C. for 2 hours to obtain a piperazine compound (BTM piperazine). Obtainable.

(Synthesis Example 5)
Exemplified compound (2-34) is synthesized according to Synthesis Example 2. Then, using the exemplified compound (2-34) as a starting material, various alkylzinc iodide reagents can be reacted in the presence of a palladium catalyst to obtain an alkyl ketone body (BTM ketone).
Powdered zinc is activated by stirring in 1N hydrochloric acid at room temperature for 4 hours, filtered, and dried overnight using a desiccator. To the tetrahydrofuran suspension of zinc thus obtained, CH ₂ Br ₂ is added dropwise at room temperature under a nitrogen atmosphere, heated to 50 ° C. and stirred for 1 minute. The reaction is cooled to room temperature and trimethylsilyl chloride (TMSCl) is added dropwise, followed by the dropwise addition of various alkyl halide tetrahydrofuran solutions. Thereafter, the temperature is raised to 50 ° C. and stirred overnight to obtain a 2.0 M alkylzinc iodide tetrahydrofuran solution.
Pd (PPh ₃ ) ₂ Cl ₂ was added to a toluene solution of Exemplified Compound (2-34), the 2.0 M alkylzinc iodide tetrahydrofuran solution was added dropwise, and the mixture was stirred at room temperature for 1 hour. BTM ketone) can be obtained.

(Synthesis Example 6)
Exemplified compound (2-25) is synthesized according to Synthesis Example 5. And propargyl hydroxyamine hydrochloride is added to the ethanol solution of exemplary compound (2-25), and it stirs for 2 hours, and can show exemplary compound (4-1).

  In a nitrogen atmosphere, boron trifluoride diethyl ether complex and 1,2-propanediol are added to a methylene chloride solution of Exemplified Compound (2-25), and the mixture is stirred at room temperature for 6 hours. Exemplified Compound (5-1) Can be obtained.

(Synthesis Example 7)
Sodium borohydride can be added to the ethanol solution of the exemplary compound (2-25) synthesized according to Synthesis Example 5 at room temperature and stirred for 2 hours to obtain the exemplary compound (5-7).
Alternatively, the exemplified compound (5-11) can be obtained by reacting a dichloromethane solution of the exemplified compound (5-7) with methoxymethyl chloride in the presence of diisopropylethylamine.
Furthermore, the pyridine solution of Exemplified Compound (5-7) can be reacted with diethyl chlorophosphate at room temperature to obtain Exemplified Compound (5-12).

(Synthesis Example 8)
Sodium borohydride can be added at room temperature to the methanol solution of the exemplary compound (2-33) synthesized according to Synthesis Example 10 described below to obtain the exemplary compound (5-14).

(Synthesis Example 9)
Sodium borohydride can be added at room temperature to the methanol solution of the exemplary compound (2-26) synthesized according to Synthesis Example 5 to obtain the exemplary compound (5-7).

(Synthesis Example 10)
A 1N lithium hydroxide solution is added to a tetrahydrofuran solution of BTMA ₂ and stirred for 30 minutes, followed by reaction for 4 hours to synthesize Example Compound (2-24). Subsequently, the reaction was performed using methoxyamine hydrochloride, 4-dimethylaminopyridine, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride in a dichloromethane solution of Exemplified Compound (2-24), and BTM64. Get. This BTM64 is dissolved in tetrahydrofuran, tri-tert-butoxyaluminolithium hydride is added, and the mixture is stirred for 1 hour to obtain the exemplary compound (2-33).

(Synthesis Example 11)
Various thiols and boron trifluoride diethyl ether complex are added to a dichloromethane solution of the exemplified compound (2-33) synthesized according to Synthesis Example 10, and the mixture is stirred overnight to obtain a thioether form (BTM thioether).
Moreover, various amines, sodium triacetoxyborohydride, and acetic acid are added to the ethanol solution of exemplary compound (2-33), and it stirs for 1 hour, An amine body (BTM amine) can be obtained.

(Synthesis Example 12)
To a methanol solution of Exemplified Compound (2-7) synthesized according to Synthesis Example 1, a methanol solution of benzyl azide compound, tris- (benzyltriazolylmethyl) amine and tetrakis (acetonitrile) copper (I) hexafluorophosphate are added, The exemplified compound (2-8) can be obtained by stirring at room temperature for 4 days.

(Synthesis Example 13)
Except for using Exemplified Compound (4-1) synthesized according to Synthesis Example 6 instead of Exemplified Compound (2-7) and using Azidoethanol instead of Benzyl Azide Compound, Exemplified Compound ( 4-4) can be obtained.

(Synthesis Example 14)
Exemplified compound (2-3) can be obtained in the same manner as in Synthetic example 13, except that exemplified compound (2-19) synthesized according to synthesis example 4 is used instead of exemplified compound (4-1).

Further, the BTMA ₂ compound of the present invention can be converted into an acid addition salt or a base addition salt according to a conventional method. Examples of the salt of the compound represented by the general formula (1) include inorganic base salts, ammonium salts, organic base salts, inorganic acid addition salts, organic acid addition salts, basic amino acid salts and the like. Examples of the inorganic base capable of generating an inorganic base salt include alkali metals (for example, sodium and potassium) and alkaline earth metals (for example, calcium). Examples of the organic base that can form an organic base salt include procaine, 2-phenylethylbenzylamine, dibenzylethylenediamine, ethanolamine, diethanolamine, trishydroxymethylaminomethane, polyhydroxyalkylamine, N-methylglucosamine, and the like. Examples of the inorganic acid that can form an inorganic acid addition salt include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Examples of the organic acid that can form an organic acid addition salt include p-toluenesulfonic acid, methanesulfonic acid, formic acid, trifluoroacetic acid, and maleic acid. Examples of basic amino acids that can form basic amino acid salts include lysine, arginine, ornithine, histidine, and the like.

The BTMA ₂ compound or salt thereof of the present invention exhibits excellent antibacterial activity and a broad antibacterial spectrum. Therefore, the BTMA ₂ compound of the present invention is useful as an antibacterial agent for pharmaceutical materials such as MRSA therapeutic agents, animal drugs, fish diseases, agricultural chemicals, food preservatives, dyes, pigments, paints, and fibers. In the present invention, the “antibacterial agent” refers to an agent that can kill a specific microorganism or weaken its activity.

  When the compound represented by the general formula (1) or a salt thereof is used as a pharmaceutical, formulation adjuvants such as excipients, carriers, and diluents that are usually used for formulation may be appropriately mixed. These are tablets, capsules, powders, syrups, granules, pills, suspensions, emulsions, solutions, powder formulations, suppositories, eye drops, nasal drops, ear drops, It can be administered orally or parenterally in the form of a patch, ointment or injection. In addition, the administration method, the dosage, and the number of administrations can be appropriately selected according to the age, weight and symptoms of the patient.

  EXAMPLES Hereinafter, although this invention is demonstrated in detail based on an Example, this invention is not limited to these.

Production Example 1 Synthesis of 55-demethoxy55-hydrazinobotromycin A ₂ (BTM1)

Under a nitrogen atmosphere, hydrazine (1.6 mL) was added to a solution of botromycin A ₂ (500 mg, 608 μmol) in ethanol (30.4 mL), and the mixture was heated to reflux at 90 ° C. for 4 hours. After the reaction solution was concentrated, the crude product was dissolved in ethyl acetate (60 mL), and washed 3 times with purified water (70 mL) and once with saturated brine (60 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated to obtain white powder of 55-demethoxy55-hydrazinobotromycin A ₂ (BTM1) (486 mg, 591 μmol) in a yield of 97%.

IR (KBr) νcm ⁻¹ : 3325 (m), 2966 (m), 2881 (w), 1653 (s), 1533 (m), 1506 (m), 1257 (w)
¹ H-NMR (300 MHz, CDCl ₃ ) δ (ppm): 0.77 (3H, d, J = 6.2 Hz), 0.80 (3H, d, J = 6.0 Hz), 0.85 to 1.14 (21 H, m), 1.41 ( 3H, d, J = 6.9Hz), 1.47 to 1.66 (2H, m), 1.91 to 1.99 (2H, m), 2.25 to 2.36 (1H, m), 2.72 to 2.88 (1H, m), 2.94 (2H, dd, J = 4.0, 14.6Hz), 3.24-3.74 (5H, m), 3.83 (2H, br, s), 3.89 (1H, s), 4.57 (1H, d, J = 10.6Hz), 4.96 (1H , Dd, J = 4.0, 9.4Hz), 5.44 (1H, br), 6.79 (1H, d, J = 9.1Hz), 7.14, (1H, d, J = 3.2Hz), 7.28-7.34 (5H, m ), 7.55 (1H, d, J = 3.5Hz), 7.76 (1H, d, J = 5.5Hz)
¹³ C-NMR (67.5 MHz, CDCl ₃ ) δ (ppm): 8.2, 11.7, 12.7, 13.1, 20.0, 20.8, 20.9, 23.1, 26.1, 28.2, 31.3, 31.5, 35.2, 39.9, 41.0, 43.6, 46.1, 49.7, 58.4, 61.5, 63.4, 112.2, 119.8, 121.1, 121.5, 133.7, 135.1, 149.6, 161.7, 162.0, 164.1, 165.3, 166.0, 166.2, 167.0
HRFABMS: calcd for C ₄₁ H ₆₂ O ₆ N ₁₀ S: 823.4653 [M + H], found m / z 823.4686 [M + H] ⁺

Production Example 2 Synthesis of Exemplary Compound (2-1) (55-benzylamino-55-demethoxybotromycin A ₂ )

Under nitrogen atmosphere at 0 ° C, BTM1 (30.2mg, 36.7μmol) N-methyl-2-pyrrolidinone (367μL) with t-butyl nitrate (20.4μL, 110μmol) and 4N dioxane hydrochloride solution (91.8μL, 367μmol) Added and stirred for 30 minutes. After 30 minutes, triethylamine (61.6 μL, 440 μmol) and benzylamine (12.0 μL, 110 μmol) were added, and the mixture was heated to 60 ° C. and stirred for 1 hour. Ethyl acetate (3 mL) was added to the reaction solution to stop the reaction, and water (3 mL) was added to separate into an organic layer. The aqueous layer was extracted 3 times with EtOAc (3 mL) and the combined organic layers were washed 5 times with water (9 mL). This organic layer was dried over sodium sulfate, filtered, and concentrated, and the crude product obtained by concentration was separated and purified by silica gel chromatography (CHCl ₃ : MeOH = 9: 1) to give an exemplary compound (2- 1) (55-benzylamino-55-demethoxybotromycin A ₂ ) (22.8 mg, 25.5 μmol) was obtained in a yield of 69%.
HRFABMS: calcd for C ₄₈ H ₆₈ O ₆ N ₉ S: 898.5056 [M + H], found m / z 898.5100 [M + H] ⁺

Production Example 3 Synthesis of Exemplary Compound (2-2) (55-demethoxy-55-morpholinobotromycin A ₂ )

By reacting and purifying BTM1 (15.0 mg, 18.2 μmol) and morpholine (20 μL, 229 μmol) in the same manner as in Production Example 2, the exemplified compound (2-2) (55-demethoxy-55-morpholinovo in the form of white powder) is purified. Thromycin A ₂ ) (11.8 mg, 13.4 μmol) was obtained with a yield of 78%.
LCMS (ESI): calcd for C ₄₅ H ₆₇ O ₇ N ₉ S: 879.13 [M + H], found m / z [M + H] ⁺

Production Example 4 Synthesis of Exemplary Compound (2-3) (55-Amino-55-demethoxybotromycin A ₂ )

By reacting and purifying BTM1 (30.0 mg, 36.4 μmol) and ammonium chloride (10 mg, 187 μmol) in the same manner as in Production Example 2, the exemplified compound (2-3) (55-amino-55-demethoxy) in the form of a white powder was purified. Bothromycin A ₂ ) (22.0 mg, 24.9 μmol) was obtained in 67% yield.
LCMS (ESI): calcd for C ₄₁ H ₆₁ O ₆ N ₉ S: 808.45 [M + H], found m / z 808.3 [M + H] ⁺

Production Example 5 Synthesis of Exemplified Compound (2-4) (55- (2-N-acetylethylene-1,2-diamino) -55-demethoxybotromycin A ₂ )

By reacting and purifying BTM1 (30.0 mg, 36.4 μmol) and N-acetylethylenediamine (18.6 μL, 182 μmol) in the same manner as in Production Example 2, the exemplified compound (2-4) (55- (2- N-acetylethylene-1,2-diamino) -55-demethoxybotromycin A ₂ ) (16.0 mg, 17.9 μmol) was obtained with a yield of 50%.
LCMS (ESI): calcd for C ₄₅ H ₆₈ O ₇ N ₁₀ S: 893.5 [M + H], found m / z 893.5 [M + H] ⁺

Production Example 6 Synthesis of Exemplary Compound (2-5) (55-demethoxy-55-thiazolidinobotromycin A ₂ )

By reacting and purifying BTM1 (30.0 mg, 36.5 μmol) and thiazolidine (14.4 μL, 182 μmol) in the same manner as in Production Example 2, the exemplified compound (2-5) (55-demethoxy-55-thiazoli) in the form of white powder is purified. Dinobotromycin A ₂ ) (22.0 mg, 24.9 μmol) was obtained in 67% yield.
LCMS (ESI): calcd for C ₄₄ H ₆₅ O ₆ N ₁₀ S ₂ : 880.45 [M + H], found m / z 880.4 [M + H] ⁺

Production Example 7 Synthesis of Exemplary Compound (2-6) (55-demethoxy-55-thiomorpholinobotromycin A ₂ )

By reacting and purifying BTM1 (30.0 mg, 36.5 μmol) and thiomorpholine (17.6 μL, 185 μmol) in the same manner as in Production Example 2, the exemplified compound (2-6) (55-demethoxy-5-55- Thiomorpholinobotromycin A ₂ ) (18.5 mg, 20.6 μmol) was obtained with a yield of 56%.
LCMS (ESI): calcd for C ₄₅ H ₆₇ O ₆ N ₉ S ₂ : 894.47 [M + H], found m / z 894.4 [M + H] ⁺

Production Example 8 Synthesis of Exemplary Compound (2-7) (55-demethoxy-55-propargylaminobotromycin A ₂ )

By reacting and purifying BTM1 (101 mg, 122 μmol) and propargylamine (23.4 μL, 364 μmol) in the same manner as in Production Example 2, the exemplified compound (2-7) (55-demethoxy-55-propargylaminoborate) in the form of a yellow powder was purified. Thromycin A ₂ ) (80.3 mg, 94.9 μmol) was obtained with a yield of 78%.
Rf: 0.57 (Silica gel, CHCl ₃ : MeOH = 9: 1)
IR (KBr) νcm ⁻¹ : 3300 (m), 2966 (m), 2881 (w), 1658 (s), 1537 (m), 1510 (m), 1257 (w)
¹ H-NMR (270 MHz, CDCl ₃ ) δ (ppm): 0.73 (6H, d, J = 4.1 Hz), 0.96 to 0.98 (18H, br), 1.05 (3H, d, J = 6.8 Hz, 8-H ), 1.43 (3H, d, J = 6.5Hz), 1.58 (2H, t, J = 10.5Hz), 1.95 to 2.07 (2H, m), 2.31 to 2.40 (1H, m), 2.73 to 2.78 (1H, m), 2.90 to 3.80 (8H, m), 3.89 (2H, br), 4.08 (1H, br), 4.51 (1H, d, J = 10.5Hz), 4.94 (1H, dd, J = 3.4, 4.8Hz ), 5.54 (1H, br), 6.88 (1H, d, J = 9.7Hz), 7.09 (1H, d, J = 3.2Hz), 7.13-7.42 (5H, m), 7.53 (1H, d, J = 3.5Hz), 7.87 (1H, d, J = 5.7Hz)
¹³ C-NMR (67.5 MHz, CDCl ₃ ) δ (ppm): 8.4, 11.2, 12.6, 13.0, 19.9, 20.8, 21.0, 22.1, 23.2, 27.3, 28.3, 31.4, 33.5, 36.1, 39.9, 42.0, 43.0, 47.2, 50.8, 58.3, 61.3, 63.6, 64.1, 72.8, 112.4, 119.8, 121.1, 121.6, 133.4, 135.1, 149.7, 161.7, 162.4, 164.1, 164.8, 165.5, 166.1, 167.1
HRFABMS: calcd for C ₃₉ H ₆₄ O ₈ N ₁₁ S: 846.4700 [M + H], found m / z 846.4731 [M + H] ⁺

Production Example 9 Synthesis of Exemplary Compound (2-8) (55-demethoxy-55- [1- (3-pyridylmethyl) -1,2,3-triazol-4-yl-4-methylamino] bothromycin A ₂ )

BTM8 (3.1mg, 3.7μmol) in methanol solution (71μl), 0.1M methanol solution of 3-azidopyridine (35μL), tris- (benzyltriazolylmethyl) amine and tetrakis (acetonitrile) copper (I) hexafluoro A mixed solution of phosphate in 0.01M MeOH (7 μL) was added and stirred at room temperature for 4 days. After confirming that the reaction was stopped by LC / MS, each reaction solution was concentrated and exemplified compound (2-8) (55-demethoxy-55- [1- (3-pyridylmethyl) -1,2, 3-Triazol-4-yl-4-methylamino] bothromycin A ₂ ) (3.5 mg) was obtained.
LCMS (ESI): Rt = 4.92min
LCMS (ESI): C ₄₉ H ₆₇ O ₆ N ₁₃ S: 980.52 [M + H], found m / z 966 [M + H] ⁺

Production Example 10 Exemplified Compound (2-9) (55- [1- (p-Bromophenyl) -1,2,3-triazol-4-yl-4-methylamino] -55-demethoxybotromycin A ₂ ) Synthesis

A methanol solution (71 μl) of Exemplified Compound (2-7) (3.1 mg, 3.7 μmol) and a 0.1 M methanol solution of 1-azido-4-bromobenzene were reacted in the same manner as in Production Example 9 to give Exemplified Compound (2-9) ) (55- [1- (p-bromophenyl) -1,2,3-triazol-4-yl-4-methylamino] -55-demethoxybotromycin A ₂ ) (3.2 mg) was obtained.
LCMS (ESI): Rt = 4.92min
LCMS (ESI): calcd for C ₅₀ H ₆₇ O ₆ BrN ₁₂ S: 1043.42 [M + H], found m / z 1042 [M + H] ⁺

Production Example 11 Exemplified Compound (2-10) (55- [1- (7-Chloro-4-isoquinolyl) -1,2,3-triazol-4-yl-4-methylamine] -55-demethoxybotro Synthesis of mycin A ₂ )

A methanol solution (71 μl) of Exemplified Compound (2-7) (3.1 mg, 3.7 μmol) and a 0.1 M methanol solution of 7-chloro-4-azidoisoquinoline were reacted in the same manner as in Production Example 9 to give Exemplified Compound (2-10 ) (55- [1- (7-chloro-4-isoquinolyl) -1,2,3-triazol-4-yl-4-methylamine] -55-demethoxybotromycin A ₂ ) (3.8 mg) Obtained.
LCMS (ESI): Rt = 5.38min
LCMS (ESI): calcd for C ₅₃ H ₆₈ O ₆ ClN ₁₃ S: 1050.48 [M + H], found m / z 1050 [M + H] ⁺

Production Example 12 Synthesis of Exemplary Compound (2-11) (55-demethoxy-55- [1- (4-pentanonyl) -1,2,3-triazol-4-yl-4-methylamino] bothromycin A ₂ )

Example Compound (2-7) (3.1 mg, 3.7 μmol) in methanol (71 μl) and 5-azido-2-pentanone in 0.1 M methanol solution were reacted in the same manner as in Production Example 9 to give Example Compound (2-11) (55-demethoxy-55- [1- (4-pentanonyl) -1,2,3-triazol-4-yl-4-methylamino] bothromycin A ₂ ) (3.6 mg) was obtained.
LCMS (ESI): Rt = 4.72min
LCMS (ESI): calcd for C ₄₉ H ₇₂ O ₇ N ₁₂ S: 973.54 [M + H], found m / z 973 [M + H] ⁺

Production Example 13 Synthesis of Exemplary Compound (2-12) (55-demethoxy-55- [1- (2-naphthalenyl) -1,2,3-triazol-4-yl-4-aminomethyl] bothromycin A ₂ )

Exemplified compound (2-7) (3.1 mg, 3.7 μmol) in methanol (71 μl) and 2- (azidomethyl) naphthalene in 0.1 M methanol were reacted in the same manner as in Production Example 9 to give exemplified compound (2-12) ( 55-demethoxy-55- [1- (2-naphthalenyl) -1,2,3-triazol-4-yl-4-aminomethyl] bothromycin A ₂ ) (3.8 mg) was obtained.
LCMS (ESI): Rt = 5.67min
LCMS (ESI): calcd for C ₅₅ H ₇₂ O ₆ N ₁₂ S: 1029.54 [M + H], found m / z 1029 [M + H] ⁺

Production Example 14 Synthesis of Exemplary Compound (2-13) (55- (1-Benzyl-1,2,3-triazol-4-yl-4-methylamine) -55-demethoxybotromycin A ₂ )

Exemplified Compound (2-7) BTM8 (3.1 mg, 3.7 μmol) in methanol (71 μl) and azidobenzene in 0.1 M methanol were reacted in the same manner as in Example 5 to give Exemplified Compound (2-13) (55- ( 1-Benzyl-1,2,3-triazol-4-yl-4-methylamine) -55-demethoxybotromycin A ₂ ) (3.6 mg) was obtained.
IR (KBr) νcm ⁻¹ : 3290 (m), 2964 (m), 2879 (w), 1658 (s), 1539 (m), 1510 (m), 1254 (w), 1232 (w)
LCMS (ESI): Rt = 5.22min
LCMS (ESI): calcd for C ₅₁ H ₇₀ O ₆ N ₁₂ S: 979.53 [M + H], found m / z 979 [M + H] ⁺

Production Example 15 Synthesis of Exemplary Compound (2-14) (55-demethoxy-55- (4-methylpiperazino) -botromycin A ₂ )

BTM1 (30.7 mg, 37.3 μmol) and 1-methylpiperazine (22.2 μL, 186 μmol) were reacted in the same manner as in Production Example 2 and purified to give the exemplified compound (2-14) (55-demethoxy-55) in the form of a white powder. -(4-Methylpiperazino) -botromycin A ₂ ) (10.6 mg, 11.9 μmol) was obtained in a yield of 32%.
LCMS (ESI): calcd for C ₄₆ H ₆₀ O ₆ N ₁₀ S ₁ : 891.52 [M + H], found m / z 891.2 [M + H] ⁺

Production Example 16 Synthesis of Exemplary Compound (2-15) (55-demethoxy-55- (4-phenylpiperazino) -botromycin A ₂ )

BTM1 (114.5 mg, 139.1 μmol) and 1-phenylpiperazine (105.7 μL, 696 μmol) were reacted in the same manner as in Production Example 2 and purified to give the exemplified compound (2-15) (55-demethoxy-55) in the form of a white powder. -(4-Phenylpiperazino) -botromycin A ₂ ) (62.2 mg, 65.2 μmol) was obtained in a yield of 47%.
LCMS (ESI): calcd for C ₅₁ H ₇₂ O ₆ N ₁₀ S: 953.54 [M + H], found m / z 952.5 [M + H] ⁺

Production Example 17 Synthesis of Exemplary Compound (2-16) (55- (4-tert-Butoxycarbonylpiperazino) -55-demethoxybotromycin)

By reacting and purifying BTM1 (151.4 mg, 183.9 μmol) and 1-tert-butoxycarbonylpiperazine (68.5 mg, 368 μmol) in the same manner as in Production Example 2, the exemplified compound (2-16) (55- (4-tert-Butoxycarbonylpiperazino) -55-demethoxybotromycin) (134.8 mg, 137.9 μmol) was obtained in 75% yield.
LCMS (ESI): calcd for C ₅₀ H ₇₇ O ₈ N ₁₀ S: 977.5647 [M + H], found m / z 977.56 [M + H] ⁺

Production Example 18 Synthesis of Exemplary Compound (2-17) (55-demethoxy-55-piperazinobotromycin A ₂ )

Trifluoroacetic acid (5.44 ml, 5.44 μmol) was added to a dichloromethane solution (520 μl) of Exemplified Compound (2-16) (50.8 mg, 52.0 μmol) at 0 ° C. in a nitrogen atmosphere and stirred for 2 hours. After stirring, the reaction solution was concentrated to obtain a white powdery exemplified compound (2-17) (55-demethoxy-55-piperazinobotromycin A ₂ ) as a mixture.
LCMS (ESI): calcd for C ₄₅ H ₆₈ O ₆ N ₁₀ S: 877.5 [M + H], found m / z 877.3 [M + H] ⁺

Production Example 19 Synthesis of Exemplary Compound (2-18) (55- (4-Benzylpiperazino) -55-demethoxybotromycin A ₂ )

Dimethyl salt of Exemplified Compound (2-17) (generated by deprotecting t-ert-butoxycarbonyl group of Exemplified Compound (2-17) with hydrochloric acid) (32.3 mg, 34 μmol) at room temperature under nitrogen atmosphere To the formamide solution (340 μl) were added potassium carbonate (18.8 mg, 136.0 μmol) and benzyl bromide (12.1 μl, 102 μmol), and the mixture was heated to 40 ° C. and stirred for 1.5 hours. After stirring, ethyl acetate (5 ml) was added, and the organic layer was washed 3 times with purified water (5 ml) and once with saturated brine (5 ml). The organic layer was dried over sodium sulfate, filtered, and concentrated, and the resulting crude product was separated and purified by silica gel chromatography (chloroform: methanol = 9: 1), and exemplified compound (2-18) (55- (4 -Benzylpiperazino) -55-demethoxybotromycin A ₂ ) (23.3 mg, 24.1 μmol) was obtained in 71% yield.
LCMS (ESI): calcd for C ₄₅ H ₆₈ O ₆ N ₁₀ S: 877.5 [M + H], found m / z 877.3 [M + H] ⁺

Production Example 20 Synthesis of Exemplary Compound (2-19) (55-demethoxy-55- (4-propargylpiperazino) bothromycin A ₂ )

Hydrochloride of Exemplified Compound (2-17) (produced by deprotecting tert-butoxycarbonyl of Exemplified Compound (2-17) with hydrochloric acid) (37.5 mg, 42.7 μmol) and 3-bromopropylene (9.5 μl, 128 μmol) ) Is reacted in the same manner as in Production Example 19 and purified to give Example Compound (2-19) (55-demethoxy-55- (4-propargylpiperazino) bothromycin A ₂ ) (26.4 mg, 28.8 μmol) was obtained with a yield of 68%.
LCMS (ESI): calcd for C ₄₈ H ₇₀ O ₆ N ₁₀ S: 915.52 [M + H], found m / z 915.5 [M + H] ⁺

Production Example 21 Synthesis of Exemplary Compound (2-20) (55-demethoxy-55- (4-methylacetate piperazino) -botromycin A ₂ )

Dimethylformamide solution of hydrochloride of Example Compound (2-17) (produced by deprotecting tert-butoxycarbonyl of Example Compound (2-17) with hydrochloric acid) at room temperature under nitrogen atmosphere (22.9 mg, 24.1 μmol) Methyl bromoacetate (6.7 μl, 72 μmol) and triethylamine (26.9 μl, 193 μmol) were added to (241 μl) and stirred for 4 hours. After stirring, ethyl acetate (5 ml) was added, and the organic layer was washed 3 times with purified water (5 ml) and once with saturated brine (5 ml). The organic layer was dried over sodium sulfate, filtered, and concentrated, and the crude product obtained by concentration was separated and purified by silica gel chromatography (chloroform: methanol = 9: 1) to give the exemplified compound (2-20) in the form of a white powder. ) (55-demethoxy-55- (4-methylacetate piperazino) -botromycin A ₂ ) (11.5 mg, 12.3 μmol) was obtained in 51% yield.
LCMS (ESI): calcd for C ₄₈ H ₇₃ O ₈ N ₁₀ S: 949.5334 [M + H], found m / z 949.53 [M + H] ⁺

Production Example 22 Synthesis of Exemplary Compound (2-21) (55- (4-Benzylacetate Piperazino) -55-demethoxybotromycin A ₂ )

Hydrochloride of Exemplified Compound (2-17) (produced by deprotecting tert-butoxycarbonyl of Exemplified Compound (2-17) with hydrochloric acid) (53.9 mg, 51.4 μmol) and benzyl 2-bromoacetate (24.4 μl, 154 μmol) was reacted in the same manner as in Production Example 21 and purified to give exemplified compound (2-21) (55- (4-benzylacetate piperazino) -55-demethoxybotromycin A _{2 in the} form of white powder. ) (21 mg, 20.5 μmol) was obtained in 40% yield.
LCMS (ESI): calcd for C ₄₈ H ₇₃ O ₈ N ₁₀ S: 1025.56 [M + H], found m / z 1025.5 [M + H] ⁺

Production Example 23 Synthesis of Exemplary Compound (2-22) (55- (4-Carboxymethylpiperazino) -55-demethoxybotromycin A ₂ )

Acetic acid ester solution (450 μl) of Exemplified compound (2-21) (20 mg, 19.5 μmol) to a suspension of 10% Pd / C (125.3 mg, 117 μmol) added to acetic acid ester (1500 μl) at room temperature under nitrogen atmosphere Was added and replaced with hydrogen, followed by stirring for 4 hours. After stirring, the reaction solution was filtered through Celite and concentrated, and then the resulting crude product was added with ethyl acetate (3 ml) and washed with purified water (3 ml). The organic layer was dried over sodium sulfate, filtered, and concentrated to give Example Compound (2-22) (55- (4-carboxymethylpiperazino) -55-demethoxybotromomycin A ₂ ) (13.6 mg, 14.6 μmol) was obtained with a yield of 75%.
HRFABMS: calcd for C ₄₇ H ₇₀ O ₈ N ₁₀ S: 957.4997 [M + Na], found m / z 957.5000 [M + Na] ⁺

Production Example 24 Synthesis of Exemplary Compound (2-23) (55-demethoxy-55-{[4- (2-hydroxyethyl) -1,2,3-4-yne] -4-methylpiperazino} bothromycin A ₂ )

At room temperature, a methanol solution (71 μl) of Exemplified Compound (2-19) (3.1 mg, 3.7 μmol) and a 0.1 M methanol solution of 2-azido-1-ethanol were reacted in the same manner as in Production Example 5 to give Exemplified Compound (2 -23) (55-demethoxy-55-{[4- (2-hydroxyethyl) -1,2,3-4-yne] -4-methylpiperazino} bothromycin A ₂ ) (3.7 mg) was obtained.
LCMS (ESI): calcd for C ₅₀ H ₇₆ O ₇ N ₁₀ S: 1003.57 [M + H], found m / z 1002.5 [M + H] ⁺

Production Example 25 Synthesis of Exemplary Compound (3-1) (54-demethoxycarbonyl-54-[(N-isopropylpropacarbamoyl) amino] bothromycin A ₂ )

Under a nitrogen atmosphere at 0 ° C., 4N hydrochloric acid dioxane solution (109 μL, 437 μmol) and n-butyl nitrate (20 μL, 109 μmol) were added to an N, N-dimethylformamide solution (365 μL) of BTM1 (30 mg, 36.5 μmol), 30 Stir for minutes. After 30 minutes, triethylamine (66 μL, 474 μmol) was added to make the reaction basic. (3 mL) was added to the reaction solution to stop the reaction, and purified water (3 mL) was added to separate into an organic layer. The aqueous layer was extracted 3 times with ethyl acetate (3 mL), and the combined organic layers were washed 5 times with water (9 mL). The organic layer was dried over sodium sulfate and concentrated to obtain a mixture of isocyanate and azide ester (24.7 mg). Under a nitrogen atmosphere, toluene (2 mL) was added to the crude product, and the mixture was stirred at 60 ° C. for 2 hours. After 2 hours, the reaction solution was cooled to room temperature, isopropylamine (15.7 μL, 182 μmol) was added, and the mixture was heated to 60 ° C. and stirred for 4 hours. Ethyl acetate (3 mL) was added to the reaction solution to stop the reaction, and purified water (3 mL) was added to separate into an organic layer. The aqueous layer was extracted 3 times with ethyl acetate (3 mL), and the combined organic layers were washed 5 times with purified water (9 mL). The organic layer was dried over sodium sulfate and concentrated, and the resulting crude product was separated and purified by silica gel chromatography (CHCl ₃ : MeOH = 10: 1) to give the exemplified compound (3-1) (white powder) ( 54-demethoxycarbonyl-54-[(N-isopropylpropacarbamoyl) amino] bothromycin A ₂ ) (13.0 mg, 15.0 μmol) was obtained in 41% yield.
LCMS (ESI): calcd for C ₄₄ H ₆₈ O ₆ N ₁₀ S: 865.5 [M + H], found m / z 865.5 [M + H] ⁺

Production Example 26 Synthesis of Exemplary Compound (3-2) (54- (N-Benzylcarbamoyl) amino-54-demethoxycarbonylbothromycin A ₂ )

By reacting BTM1 (30.0 mg, 36.5 μmol) and benzylamine (25 μL, 24 μmol) in the same manner as in Production Example 25 and purifying, exemplary compound (3-2) (54- (N-benzylcarbamoyl) in the form of white powder Amino-54-demethoxycarbonylbothromycin A ₂ ) (6.3 mg, 17.9 μmol) was obtained in 50% yield.
LCMS (ESI): calcd for C ₄₈ H ₆₈ O ₆ N ₁₀ S: 913.5 [M + H], found m / z 913.5 [M + H] ⁺

Production Example 27 Synthesis of Exemplary Compound (3-3) (54-demethoxycarbonyl-54- [N- (3-thiazocarbonyl) amino] bothromycin A ₂ )

By reacting BTM1 (30.6 mg, 37.2 μmol) and thiazolidine (6.6 μL, 84 μmol) in the same manner as in Production Example 25 and purifying, exemplary compound (3-3) (54-demethoxycarbonyl-54- [N- (3-thiazocarbonyl) amino] bothromycin A ₂ ) (4.3 mg, 4.8 μmol) was obtained in a yield of 13%.
LCMS (ESI): calcd for C ₄₄ H ₆₆ O ₆ N ₁₀ S ₂ : 895.46 [M + H], found m / z 895.2 [M + H] ⁺

Production Example 28 Synthesis of Exemplary Compound (3-4) (54- [N- (4-acetylaminoethylenecarbonyl) amino] -54-demethoxycarbonylbotromycin A ₂ )

By reacting and purifying BTM1 (30.0 mg, 36.5 μmol) and N-acetylethylenediamine (10.8 mg, 106 μmol) in the same manner as in Production Example 25, the exemplified compound (3-4) (54- [N- (4-Acetylaminoethylenecarbonyl) amino] -54-demethoxycarbonylbotromycin A ₂ ) (7.6 mg, 8.3 μmol) was obtained in a yield of 23%.
LCMS (ESI): calcd for C ₄₅ H ₆₉ O ₇ N ₁₁ S: 908.51 [M + H], found m / z 908.3 [M + H] ⁺

Production Example 29 Synthesis of Exemplary Compound (2-24) (55-demethoxy-55-hydroxybothromycin A ₂ )

Under a nitrogen atmosphere, 1N lithium hydroxide aqueous solution (510 μL, 510 μmol) was added to a trihydrofuran solution (4.2 mL) of botromycin A ₂ (350 mg, 425 μmol) at room temperature, and the mixture was stirred for 30 minutes. After 4 hours, the reaction solution was concentrated to obtain Exemplified Compound (2-24) (55-demethoxy-55-hydroxybothromycin A ₂ ) (534.7 mg) as a white powder.
Rf = 0.03 (Silica gel, CHCl ₃ : MeOH = 9: 1)
IR (KBr) νcm ⁻¹ : 3317 (m), 2972 (m), 1672 (s), 1522 (m), 1419 (w), 1196 (s), 1144 (m)
¹ H-NMR (300 MHz, CDCl ₃ ) δ (ppm): 0.87 (3H, d, J = 6.8 Hz), 0.91 to 1.04 (18H, br), 1.26 (3H, d, J = 7.2 Hz), 1.80 to 1.96 (2H, m), 2.08 to 2.24 (3H, m), 3.81 (1H, s), 3.88 to 4.34 (6H, m), 4.59 (1H, d, J = 17.9Hz), 4.57 (1H, d, J = 10.8Hz), 4.76 (1H, m), 3.05 to 3.25 (4H, m), 5.70 to 5.80 (1H, m), 7.09 to 7.40 (5H, m), 7.52 (1H, d, J = 3.6Hz ), 7.73 (1H, d, J = 3.0Hz)
HRFABMS: calcd for C ₄₁ H ₆₁ O ₇ N ₈ S: 809.4384 [M + H], found m / z 809.4390 [M + H] ⁺

Production Example 30 Synthesis of Exemplary Compound (2-34) (55-demethoxy-55-ethylthiobothromycin A ₂ )

In a nitrogen atmosphere at 0 ° C, BTM1 (30.0 mg, 36.4 μmol) in a mixed solution of tetrohydrofuran (728 μl) dimethylformamide (600 μL) with 4N dioxane hydrochloride (91.1 μL, 364.2 μmol), t-butyl nitrate (20.2 μL) , 109.2 μmol), and stirred for 1 hour. After 1 hour, triethylamine (61.1 μL, 436.8 μmol) and ethyl mercaptan (8.0 μL, 109.2 μmol) were added, and the mixture was warmed to room temperature and stirred overnight. Ethyl acetate was added to the reaction solution to stop the reaction, and water was added to separate into an organic layer. The aqueous layer was extracted twice with ethyl acetate, and the combined organic layers were washed 5 times with water. This organic layer was dried over sodium sulfate, filtered, and concentrated, and the crude product obtained by concentration was separated and purified by silica gel chromatography (CHCl ₃ : MeOH = 9: 1) to give an exemplary compound (2- 34) (55-demethoxy-55-ethylthiobothromycin A ₂ ) (25.3 mg, 29.6 μmol) was obtained in 81% yield.
HRFABMS: calcd for C ₄₃ H ₆₅ O ₆ N ₈ S ₂ : 853.4469 [M + H], found m / z 853.4457 [M + H] ⁺

Production Example 31 Synthesis of Exemplified Compound (2-35) (55-demethoxy-55-propylthiobothromycin A ₂ )

BTM1 (20.0 mg, 24.3 μmol) and 1-propanethiol (11 μL, 122 μmol) were reacted in the same manner as in Production Example 30 to give exemplary compound (2-35) (55-demethoxy-55-propylthiobotro) in the form of a yellow powder. Mycin A ₂ ) (3.0 mg, 3.46 μmol) was obtained in 14% yield.
LCMS (ESI): calcd for C ₄₄ H ₆₆ O ₆ N ₈ S: 867.45 [M + H], found m / z 867.3 [M + H] ⁺

Production Example 32 Synthesis of Exemplary Compound (2-36) (55-demethoxy-55-isopropylthiobothromycin A ₂ )

BTM1 (30.0 mg, 36.5 μmol) and 2-propanol (17 μL, 182 μmol) were reacted in the same manner as in Production Example 30 and purified to give the exemplified compound (2-36) (55-demethoxy-55-isopropyl) as a white powder. Thiobotromycin A ₂ ) (8.3 mg, 9.5 μmol) was obtained in a yield of 26%.
LCMS (ESI): calcd for C ₄₄ H ₆₆ O ₆ N ₈ S ₂ : 867.45 [M + H], found m / z 867.4 [M + H] ⁺

Production Example 33 Synthesis of Exemplified Compound (2-37) (55-[(3-acetylamino) ethylthio] -55-demethoxybotromycin)

By reacting and purifying BTM1 (30.0 mg, 36.5 μmol) and N-acetylcysteamine (19 μL, 179 μmol) in the same manner as in Production Example 30, the exemplified compound (2-37) (55-[(3- Acetylamino) ethylthio] -55-demethoxybotromycin) (23.1 mg, 25.3 μmol) was obtained in 70% yield.
HRFABMS: calcd for C ₄₅ H ₆₈ O ₇ N ₉ S ₂ : 910.4680 [M + H], found m / z 910.4700 [M + H] ⁺

Production Example 34 Synthesis of Exemplary Compound (2-38) (55-demethoxy-55-[(4-methoxycarbonyl) ethylthio] bothromycin A ₂ )

By reacting and purifying BTM1 (30.0 mg, 36.5 μmol) and methyl N-3-mercaptopropionate (18 μL, 182 μmol) in the same manner as in Production Example 30, the exemplified compound (2-38) (55- Demethoxy-55-[(4-methoxycarbonyl) ethylthio] bothromycin A ₂ ) (11.7 mg, 12.8 μmol) was obtained in 35% yield.
LCMS (ESI): calcd for C ₄₅ H ₆₆ O ₈ N ₈ S: 911.44 [M + H], found m / z 911.4 [M + H] ⁺

Production Example 35 Synthesis of Exemplary Compound (2-39) (55-demethoxy-55-[(3-methoxycarbonyl) methylthio] -botromycin A ₂ )

By reacting and purifying BTM1 (30.0 mg, 36.5 μmol) and methyl thioglycolate (16.3 μL, 182 μmol) in the same manner as in Production Example 30, the exemplified compound (2-39) (55-demethoxy-55) in the form of a yellow powder was purified. -[(3-Methoxycarbonyl) methylthio] -botromycin A ₂ ) (22.5 mg, 25.0 μmol) was obtained in 69% yield.
LCMS (ESI): calcd for C ₄₄ H ₆₈ O ₆ N ₁₀ S: 897.43 [M + H], found m / z 896.43 [M + H] ⁺

Production Example 36 Synthesis of Exemplary Compound (2-25) (55-demethoxy-55-propylbothromycin A ₂ )

A zinc reagent as a reaction reagent was prepared in advance. Zinc powder (1 g, 15 mmol) activated with propyl iodide (15 ml, 1.5 mmol) and dilute hydrochloric acid was heated to reflux for 2 hours in THF (15 ml) under a nitrogen atmosphere. The reaction solution was cooled to room temperature to prepare 1.0 M zinc iodide reagent.
The thiol ester exemplary compound (2-34) (30 mg, 35.0 μmol) and the dichlorobistriphenylphosphine catalyst were dissolved in THF (1 ml) at room temperature under a nitrogen atmosphere. The previously prepared zinc iodide reagent (42 μl) was added dropwise to the reaction solvent and stirred for 6 hours. After stirring, purified water was added to terminate the reaction, and the mixture was filtered through celite. The organic layer was washed with purified water (10 ml) and saturated brine (10 ml). The organic product was dried over sodium sulfate, filtered, and concentrated to give a crude product, which was separated and purified by silica gel chromatography (CHCl ₃ : MeOH = 9: 1) to give the exemplified compound (2-25) (2-25) ( 55-demethoxy-55-propylbothromycin A ₂ ) (21 mg, 25.1 μmol) was obtained in 72% yield.
¹³ C-NMR (67.5 MHz, CDCl ₃ ) δ (ppm): 13.6, 15.5, 16.1, 16.9, 19.6, 20.2, 26.9, 27.7, 21.0 (), 22.1 (C-52), 23.2, 27.3, 28.3, 31.4 , 33.5, 36.1, 39.9, 42.0, 43.0, 47.2, 50.8, 58.3, 61.3, 63.6, 64.1, 72.8, 112.4, 119.8, 121.1, 121.6, 133.4, 135.1, 149.7, 161.7, 162.4, 164.1, 164.8, 165.5, 166.1 , 167.1
¹ H-NMR (270 MHz, CDCl ₃ ) δ (ppm): 0.72 (3H, d, J = 6.5 Hz), 0.80 (3H, d, J = 6.5 Hz), 0.87 to 0.98 (24 H, br), 1.36 ( 3H, d, J = 7.3Hz), 1.56 to 1.64 (2H, m), 1.89 to 2.09 (4H, br), 2.42 (1H, t, J = 7.3Hz), 2.35 to 2.59 (1H, m), 2.70 ~ 2.89 (1H, m), 2.99 (1H, dd, J = 4.6, 18.6), 3.23 (1H, dd, J = 5.5, 18.0Hz), 3.34 to 3.45 (2H, br), 3.47 to 3.81 (4H, m), 3.91 (3H, br), 4.00 (1H, br), 4.59 (1H, d, J = 10.3Hz), 4.88 to 4.89 (1H, br), 5.54 (1H, m), 6.88 (1H, d , J = 8.9Hz), 7.18 (1H, br), 7.22 to 7.39 (12H, m), 7.55 (1H, d, J = 3.4Hz, 46-H), 7.84 (1H, d, J = 5.7Hz, 8.78 (1H, br)
HRFABMS: calcd for C ₄₄ H ₆₇ O ₆ N ₈ S: 835.4904 [M + H], found m / z 835.4924 [M + H] ⁺

Production Example 37 Synthesis of Exemplary Compound (2-26) (55-demethoxy-55-ethylbotromycin A ₂ )

Exemplified compound (2-34) (40 mg, 46.8 μmol) and 0.9M ethylzinc iodide reagent (78 ml, 70 μmol) were reacted in the same manner as in Production Example 36 and purified to give an exemplified compound (2-26 ) (55-demethoxy-55-ethylbothromycin A ₂ ) (37.0 mg, 45.1 μmol) was obtained in 96% yield.
LCMS (ESI): calcd for C ₄₃ H ₆₄ O ₆ N ₈ S: 821.47 [M + H], found m / z 821.2 [M + H] ⁺

Production Example 38 Synthesis of exemplified compound (2-27) (55-butyl-55-demethoxybotromycin A ₂ )

Exemplified compound (2-34) (30 mg, 35.0 μmol) and 1.0 M butylzinc iodide reagent (42 μl, 42 μmol) were reacted in the same manner as in Production Example 36 and purified to give an exemplified compound (2-27 ) (55-butyl-55-demethoxybotromycin A ₂ ) (18.0 mg, 21.2 μmol) was obtained in a yield of 60%.
LCMS (ESI): calcd for C ₄₅ H ₆₉ O ₆ N ₈ S: 849.5061 [M + H], found m / z 849.51 [M + H] ⁺

Production Example 39 Synthesis of Exemplified Compound (2-28) (55-demethoxy-55-isobutylbothromycin A ₂ )

Exemplified compound (2-34) (30 mg, 35.0 μmol) and 1.0 M butylzinc iodide reagent (42 μl, 42.0 μmol) were reacted in the same manner as in Production Example 36 and purified to give an exemplified compound (2- 28) (55-demethoxy-55-isobutylbotromycin A ₂ ) (14.0 mg, 16.5 μmol) was obtained in a yield of 47%.
LCMS (ESI): calcd for C ₄₅ H ₆₈ O ₆ N ₈ S: 849.50 [M + H], found m / z 849.3 [M + H] ⁺

Production Example 40 Synthesis of Exemplary Compound (2-29) (55-demethoxy-55-octylbotromycin A ₂ )

Exemplified compound (2-34) (30 mg, 35.0 μmol) and 1.0 M octylzinc iodide reagent (42 μl, 42 μmol) were reacted in the same manner as in Production Example 36 and purified to give an exemplary compound (2-29) in the form of a white powder. ) (55-demethoxy-55-octylbotromycin A ₂ ) (20.1 mg, 21.7 μmol) was obtained in 62% yield.
LCMS (ESI): calcd for C ₄₉ H ₇₆ O ₆ N ₈ S: 905.56 [M + H], found m / z 927.3 [M + H] ⁺

Production Example 41 Synthesis of Exemplary Compound (2-30) (55-demethoxy-55-isopropylbothromycin A ₂ )

Exemplified compound (2-34) (80 mg, 58.6 μmol) and 1.0 M isopropylzinc iodide reagent (58 μl, 58 μmol) were reacted in the same manner as in Production Example 36 and purified to give an exemplary compound (2-30 in the form of white powder). ) (55-demethoxy-55-isopropylbothromycin A ₂ ) (17.6 mg, 21.1 μmol) was obtained in a yield of 36%.
LCMS (ESI): calcd for C ₄₄ H ₆₆ O ₆ N ₈ S: 835.48 [M + H], found m / z 835.3 [M + H] ⁺

Production Example 42 Synthesis of Exemplified Compound (2-31) (55-benzyl-55-demethoxybotromycin A ₂ )

Exemplified compound (2-34) (20 μg, 23.4 μmol) and 0.9M benzylzinc iodide reagent (58 μl, 58 μmol) were reacted in the same manner as in Production Example 36 and purified to give an exemplary compound (2-31) in the form of a white powder. ) (55-benzyl-55-demethoxybotromycin A ₂ ) (8.9 mg, 10.1 μmol) was obtained in 43% yield.
LCMS (ESI): calcd for C ₄₈ H ₆₆ O ₆ N ₈ S: 883.48 [M + H], found m / z 883.3 [M + H] ⁺

Production Example 43 Synthesis of Exemplary Compound (2-32) (55-demethoxy-55- (3-ethoxycarbonylethyl) bothromycin A ₂ )

  Ethyl iodide propionate to be used was added with ethanol and 20 v% sulfuric acid to a benzene solution (5 ml) of iodopropionic acid (2 g, 10 mmol) in a nitrogen atmosphere and heated under reflux for 1 hour. After cooling to room temperature, the mixture was concentrated and ethyl acetate (10 ml) was added. The organic layer was washed with purified water (10 ml) and saturated brine (10 ml), dried over magnesium sulfate, filtered, and concentrated to propionate iodide. Ethyl acid was obtained quantitatively. This ethyl iodide propionate (1.71 ml, 7.5 mmol) and activated zinc powder (0.5 g, 7.5 mmol) treated with dilute hydrochloric acid were allowed to act in THF and heated under reflux for 2 hours under a nitrogen atmosphere to give 1.0 M propionated iodide. An ethyl zinc acid reagent was obtained. Thereafter, the reaction solution was cooled to room temperature.

Under a nitrogen atmosphere, dichlorobistriphenylphosphine catalyst was added to a tetrahydrofuran solution (2 ml) of Exemplified Compound (2-34) (105 mg, 123 μmmol) at room temperature, and the previously prepared ethyl zinc iodide propionate reagent (200 μl) was added dropwise. And stirred for 6 hours. After stirring, water was added to complete the reaction, and the mixture was filtered through celite and concentrated. Ethyl acetate (10 ml) was added, and the organic layer was washed with purified water (10 ml) and saturated brine (10 ml). The organic layer was dried over magnesium sulfate, filtered, and concentrated to give a crude oily product, which was separated and purified by silica gel chromatography (chloroform: methanol = 9: 1) to give the exemplified compound (2-32) (55- Demethoxy-55- (3-ethoxycarbonylethyl) bothromycin A ₂ ) (60 mg) was obtained in a yield of 55%.
HRFABMS: calcd for C ₄₆ H ₆₈ O ₈ N ₈ S: 915.4779 [M + Na], found m / z 915.4788 [M + Na] ⁺

Production Example 44 Synthesis of Exemplified Compound (4-1) (55-demethoxy-55-O-propargyloxime-55-propylbothromycin A ₂ )

Propargyl hydroxylamine hydrochloride (5.15 mg, 47.9 μmol) was added to an ethanol solution (234 μl) of Exemplified Compound (2-25) (20 mg, 23.4 μmol) at room temperature under a nitrogen atmosphere and stirred for 2 hours. After stirring, purified water was added to complete the reaction, and the mixture was concentrated. Ethyl acetate was added, and the organic layer was washed with purified water (3 ml) and saturated brine (3 ml). The organic layer was dried over sodium sulfate, filtered, and concentrated to obtain an oily composition product, which was purified by silica gel chromatography (chloroform: methanol = 9: 1) to give the exemplified compound (4-1) (55-demethoxy). -55-O-propargyloxime-55-propylbothromycin A ₂ ) (18 mg, 19.9 μmol) was obtained in 85% yield.
LCMS (ESI): calcd for C ₄₇ H ₆₉ O ₆ N ₉ S: 888.5 [M + H], found m / z 888.6 [M + H] ⁺

Production Example 45 Synthesis of Exemplary Compound (5-7) (55-demethoxy-55-hydroxy-55-propylbothromycin A ₂ )

Under a nitrogen atmosphere, sodium borohydride (2.6 mg, 67 μmol) was added to an ethanol solution (350 μl) of the exemplified compound (2-25) (30 mg, 35 μmol) at room temperature, and the mixture was stirred for 2 hours. After stirring, purified water was added to terminate the reaction. After concentration, ethyl acetate (10 ml) was added, and the organic layer was washed with purified water (10 ml) and saturated brine (10 ml). The organic layer was dried over magnesium sulfate, filtered, and concentrated, and the crude product obtained was separated and purified by silica gel chromatography (chloroform: methanol = 9: 1) to give the exemplified compound (5-7) (555-demethoxy-55 -Hydroxy-55-propylbothromycin A ₂ ) (18 mg, 22.4 μmol) was obtained in a yield of 64%.
LCMS (ESI): calcd for C ₄₄ H ₆₈ O ₆ N ₉ S: 837.5 [M + H], found m / z 837.4 [M + H] ⁺

Production Example 46 Synthesis of Exemplary Compound (5-1) (55-demethoxy-55,55- (1,3-dithiane) -55-propylbothromycin A ₂ )

To a methylene chloride solution (2 ml) of Exemplified Compound (2-25) (38 mg, 45 μmol) at −20 ° C. in a nitrogen atmosphere, boron trifluoride diethyl ether complex (12.5 μl, 99 μmol), 1,2-ethanedithiol ( 5.5 μl, 54 μmol) was added, and the temperature was raised to room temperature and stirred for 6 hours. After stirring, purified water was added to terminate the reaction, diethyl ether (5 ml) was added, washed with purified water (10 ml) and saturated brine (10 ml), dried over sodium sulfate, filtered and concentrated. The crude oily product was separated and purified by silica gel chromatography (chloroform: methanol = 9: 1), and exemplified compound (5-1) (55-demethoxy-55,55- (1,3-dithiane) -55. -Propylbotromycin A ₂ ) (23 mg, 24.8 μmol) was obtained in a yield of 55%.
HRFABMS: calcd for C ₄₇ H ₇₂ O ₅ N ₈ S ₃ : 947.4686 [M + Na], found m / z 947.4700 [M + Na] ⁺

Production Example 47 Synthesis of Exemplary Compound (2-33) (55-demethoxy-55-hydrobotromycin A ₂ )

N-[(3S, 6S, 14R, 14aS) -6-tert-Butyl-3-isopropyl-14-methyl-1,4,10-trioxododecahydropyrrolo [1,2-a] [1, 4,7,10] tetraazacyclododecin-7 (8H) -yliden] -3-methyl-L-valyl- (betas) -N-[(1R) -3- [methoxy (methyl) amino] -3-oxo-1 -(1,3-thiazol-2-yl) propyl] -beta-methyl-L-phenylalaninamid) (see WO 2006 / 103010A1 pamphlet) (76.76 mg, 89.9 μmol) in tetrahydrofuran (899 μl) solution with hydrogen Tri-tert-butoxyalumino lithium chloride (58.1 mg, 228.4 μmol) was added and stirred for 5 hours. Dichloromethane was added to the reaction solution to stop the reaction, and water was added to separate into an organic layer. This organic layer was dried over sodium sulfate, filtered, and concentrated, and the crude product obtained by concentration was separated and purified by silica gel chromatography (CHCl ₃ : MeOH = 40: 1 to 20: 1) to give an example of a white powder. Compound (2-33) (55-demethoxy-55-hydrobotromycin A ₂ ) (61.3 mg, 77.3 μmol) was obtained in a yield of 86%.
HRFABMS: calcd for C ₄₁ H ₆₁ O ₆ N ₈ S: 793.4435 [M + H], found m / z 793.4454 [M + H] ⁺

Production Example 48 Synthesis of Exemplary Compound (4-2) (55-demethoxy-55- (N-methoxy) iminobotromycin)

Methoxyamine hydrochloride (2 mg, 23.9 μmol) was added to an ethanol solution (200 μl) of Exemplified Compound (2-33) (8.5 mg, 11.6 μmol) at 0 ° C. in a nitrogen atmosphere, and the mixture was stirred for 30 minutes. After 30 minutes, the reaction solution was concentrated, dissolved in water, adjusted to neutrality by adding 1N aqueous sodium hydroxide solution, and extracted with chloroform. The organic layer was dried over sodium sulfate, filtered, and concentrated, and the crude product obtained was separated and purified by silica gel chromatography (CHCl ₃ : MeOH = 9: 1) to give an exemplary compound (4- 2) (55-demethoxy-55- (N-methoxy) iminobotromycin) (6.9 mg, 8.4 μmol) was obtained in a yield of 72%.
LCMS (ESI): calcd for C ₄₂ H ₆₃ O ₆ N ₉ S: 821.47 [M + H], found m / z 822.2 [M + H] ⁺

Preparation Example 49 Synthesis of Exemplary Compound (4-3) (55-demethoxy-55-oximbothromycin A ₂ )

Exemplified compound (2-33) (8.5 mg, 11.6 μmol) and hydroxylamine hydrochloride (4 mg, 57.6 μmol) were reacted in the same manner as in Example 48 and purified to give exemplary compound (4-3) in the form of a white powder. (55-demethoxy-55-oximbothromycin A ₂ ) (22.2 mg, 27.5 μmol) was obtained in 96% yield.
LCMS (ESI): calcd for C ₄₁ H ₆₁ O ₆ N ₉ S: 808.45 [M + H], found m / z 808.3 [M + H] ⁺

Production Example 50 Synthesis of Exemplary Compound (5-9)

2-aminoethanethiol hydrochloride (2.8 mg, 24.6 μmol), potassium carbonate in a water: ethanol mixed solution (13:30, 200 μL) of Exemplified Compound (2-33) (10 mg, 12.6 μmol) at room temperature under a nitrogen atmosphere (5.2 mg, 37.6 μmol) was added and stirred for 4 hours. After 4 hours, water was added to the reaction solution to stop the reaction, and chloroform was added to separate into an organic layer. The aqueous layer was extracted 3 times with ethyl acetate (3 mL), and the combined organic layers were washed 5 times with water (9 mL). The organic layer was washed with saturated brine, dried over sodium sulfate and concentrated.The resulting crude product was separated and purified by silica gel chromatography (chloroform: methanol = 9: 1) to give a white powder. Compound (5-9) (7.2 mg, 8.4 μmol) was obtained in a yield of 67%.
LCMS (ESI): calcd for C ₄₃ H ₆₅ O ₅ N ₉ S ₂ : 852.46 [M + H], found m / z 852.3 [M + H] ⁺

Production Example 51 Synthesis of Exemplary Compound (5-10) (54-demethoxycarbonyl-54- (4N-morpholino) bothromycin A ₂ )

In a nitrogen atmosphere at room temperature, the exemplified compound (2-33) (10 mg, 12.6 μmol) in ethanol solution (126 μL) was added to morpholine (5 μl, 63 μmol), sodium triacetoxyborohydride (4 mg, 18.9 μmol), acetic acid (20 μl, 0.35 μmol) was added and stirred for 1 hour. After stirring, chloroform was added to the reaction solution, and the organic layer was washed with a saturated aqueous bicarbonate solution and saturated brine. The organic layer was dried over sodium sulfate, filtered, and concentrated. The crude product obtained by concentration was separated and purified by silica gel chromatography (chloroform: methanol = 9: 1) to give an exemplary compound (5- 10) (54-demethoxycarbonyl-54- (4N-morpholino) bothromycin A ₂ ) (6.6 mg, 7.6 μmol) was obtained in a yield of 60%.
LCMS (ESI): calcd for C ₄₅ H ₆₉ O ₆ N ₉ S: 864.51 [M + H], found m / z 865.4 [M + H] ⁺

Production Example 52 Synthesis of Exemplary Compound (5-2) (54-demethoxycarbonyl-54,54- (1,3-dithiane) bothromycin A ₂ )

In a nitrogen atmosphere at room temperature, Exemplified Compound (2-33) (20.2 mg, 25.2 μmol) in dichloromethane solution (252 μl) was mixed with 1,3-propanedithiol (2.99 mg, 29.8 μmol), boron trifluoride diethyl ether complex (6.9 μl, 54.6 μmol) was added and stirred overnight. After stirring, chloroform (3 ml) was added, washed with water (3 ml) and saturated brine (3 ml), the organic layer was dried over sodium sulfate and filtered, and the resulting crude product was subjected to silica gel chromatography (chloroform: methanol = 9: 1), 38% of the exemplified compound (5-2) (54-demethoxycarbonyl-54- (1,3-dithiane) bothromycin A ₂ ) (8.3 mg, 9.4 μmol) in the form of a white powder was obtained by 38% The yield was obtained.
HRFABMS: calcd for C ₄₄ H ₆₇ O ₅ N ₈ S ₃ : 883.4397 [M + H], found m / z 883.4400 [M + H] ⁺

Production Example 53 Synthesis of Exemplary Compound (5-3) (54-demethoxycarbonyl-54,54- (1,2-ethanedithio) bothromycin A ₂ )

By reacting Exemplified Compound (2-33) (21.4 mg, 27.0 μmol) with 1,2-ethanedithiol (2.7 μl, 32.4 μmol) in the same manner as in Production Example 52 and purifying, Exemplified Compound in the form of white powder ( 5-3) (54-demethoxycarbonyl-54,54- (1,2-ethanedithio) bothromycin A ₂ ) (7.3 mg, 8.4 μmol) was obtained in a yield of 32%.
HRFABMS: calcd for C ₄₃ H ₆₅ O ₅ N ₈ S ₃ : 869.4240 [M + H], found m / z 869.4200 [M + H] ⁺

Production Example 54 Synthesis of Exemplary Compound (5-4) (54-demethoxycarbonyl-54,54-bis (1-ethylthionyl) bothromycin A ₂ )

Exemplified compound (2-3) (21.6 mg, 27.2 μmol) and ethanethiol (4.8 μl, 65.3 μmol) were reacted in the same manner as in Production Example 52 and purified to give exemplary compound (5-4) in the form of white powder (54-demethoxycarbonyl-54,54-bis (1-ethylthionyl) bothromycin A ₂ ) (11.4 mg, 12.7 μmol) was obtained in a yield of 47%.
HRFABMS: calcd for C ₄₅ H ₇₁ O ₅ N ₈ S ₃ : 899.4710 [M + H], found m / z 899.4700 [M + H] ⁺

Production Example 55 Synthesis of Exemplary Compound (5-5) (54-demethoxycarbonyl-54,54-bis (1-propylthionyl) bothromycin A ₂ )

Exemplified compound (2-33) (20.0 mg, 25.2 μmol) and propanethiol (5.5 μl, 60.5 μmol) were reacted in the same manner as in Production Example 52 and purified to give exemplary compound (5-5) in the form of white powder (54-demethoxycarbonyl-54,54-bis (1-propylthionyl) bothromycin A ₂ ) (14.4 mg, 15.5 μmol) was obtained in a yield of 63%.
HRFABMS: calcd for C ₄₇ H ₇₅ O ₅ N ₈ S ₃ : 927.5023 [M + H], found m / z 927.500 [M + H] ⁺

Production Example 56 Synthesis of Exemplary Compound (5-6) (54-demethoxycarbonyl-54,54-bis (3-hydroxy-1-ethylthionyl) bothromycin A ₂ )

The exemplified compound (2-33) (21.2 mg, 26.7 μmol) and 2-mercaptoethanol (2.3 μl, 32.0 μmol) were reacted in the same manner as in Production Example 52 and purified to give the exemplified compound (5- 6) (54-demethoxycarbonyl-54,54-bis (3-hydroxy-1-ethylthionyl) bothromycin A ₂ ) (6.3 mg, 6.8 μmol) was obtained in a yield of 25%.
HRFABMS: calcd for C ₄₅ H ₇₁ O ₇ N ₈ S ₃ : 931.4608 [M + H], found m / z 931.4600 [M + H] ⁺

Production Example 57 Synthesis of Exemplified Compound (5-8) (54-demethoxycarbonyl-54-hydroxy-54- (2-thiazole) bothromycin)

In a nitrogen atmosphere at 0 ° C., a solution of Exemplified Compound (2-33) (25.1 mg, 31.7 μmol) in dichloromethane (317 μl) in 2-trimethylsilylthiazole (8.1 mg, 51.5 μmol) and 1N tetrabutylammonium fluoride tetrahydrofuran solution ( 47.6 μl, 47.6 μmol) were added and stirred for 9 hours. After stirring, the reaction mixture was concentrated, separated and purified by silica gel chromatography (chloroform: methanol = 100: 1 to 40: 1), and exemplified compound (5-8) (54-demethoxycarbonyl-54- Hydroxy-54- (2-thiazole) bothromycin) (12.3 mg, 14.0 μmol) was obtained in 44% yield.
HRFABMS: calcd for C ₄₄ H ₆₄ O ₆ N ₉ S ₃ : 878.4421 [M + H], found m / z 878.4400 [M + H] ⁺

In addition, the identification of the compound was performed as follows.
Infrared (IR) absorption spectra were measured using JASCO Corporation [JASCO] FT / IR-460Plus.
¹ H-NMR spectrum using JEOL JNM-EX270 (270 MHz) spectrometer, Varian VXR-300 (300 MHz), ¹³ C-NMR using JEOL JNM-EX270 (67.5 MHz) spectrometer, Varian VXR-300 (75 MHz) It was measured. Chemical shifts are expressed in δ (ppm), and coupling patterns are indicated by the following abbreviations.
s: singlet, d: doublet, dd: double doublet, ddd: double double doublet, q: quartet, m: multiplet, br: broad

For thin layer chromatography (prep.TLC), Silicagel 60 F254 manufactured by Merck was used, and UV irradiation (254 nm) and phosphomolybdenum color were used for detection of the compound.
Column chromatography was performed by flash column chromatography, and a column packed with Merca silica gel (Art.1.09385) was used.
Preparative HPLC is HITACHI L-2130 type pump, L-2400 type detector (column PEGASIL ODS 1 20φx250mm, 35% CH ₃ CN with 0.05% TFA (solvent A) / H ₂ O 0.1% with TFA (solvent B) , Using a flow rate of 10 mL / min, detection UV of 210 nm).
Low-resolution mass spectra (LC / MS) were measured using Waters micromass ZQ and Waters 2795 Separations Module (column Shenshu Pak HPLC COLUMN PEGASIL ODS, 2.0φ × 50 mm, gradient 8 minutes to 10% solvent A / B to 100% solvent A, flow rate 0.3 High resolution mass spectrum (HR-MS) was measured using JEOL JMS-AX505 HA Mass Spectrometer using mL / min, detection Photo Diode Array (210 to 400 mm).

Test example 1
The antibacterial activity (minimum growth inhibitory concentration (MIC)) of conventional antibiotics, BTMA ₂ and the BTMA ₂ compound of the present invention against various microorganisms shown in Table 1 was measured. The minimum inhibitory concentration was measured according to the 2008 version of Clinical and Laboratory Standard Institutes (CLSI) method M7-A7. Details of the measurement method are as follows.
First, a stock strain was raised in MHB (Mullar Hinton broth) and cultured at 35 ° C. for 18 to 20 hours. Each antibiotic, BTMA ₂ and BTMA ₂ compound were dissolved in 50% methanol to 1280 μg / mL, and dilution series of 20, 40, 80, 160, 320, 640, and 1280 μg / mL were prepared by doubling dilution. 1.0 mL of the solution of each concentration was dispensed, and 9 mL of sterilized MHA (Mullar Hinton agar) was added and solidified on a plate. The cultured bacterial solution was adjusted so that the amount of bacteria was about 1.0 × 10 ⁸ CFU / mL, and 10 μL of bacterial solution and 90 μL of MHB were placed in a 27-well planter and inoculated on the agar plate. Each MIC was determined after culturing the agar plate at 35 ° C. for 18 to 20 hours. Gram-positive bacteria that are test bacteria include Stphylococcus aureus FDA209P and Smith strain, MRSA 1,2,70,92-1191 strain with multidrug resistance, and S. aureus 447,217,2104,97 strain with macrolide resistance Using. Moreover, NCTC12201 and NCTC12203 strains having E.faecalis ATCC21212 and vancomycin resistance gene (vanA) were used as enterococci. Furthermore, S. epidermidis IFO12648 and M. luteus ATCC9341 were used as other test bacteria.
The results are shown in Table 1.

As is apparent from the results in Table 1, the BTMA ₂ compound of the present invention was found to exhibit a broad antibacterial spectrum against gram-positive bacteria.

Test example 2
As an index of the stability of BTMA ₂ and each BTMA ₂ compound in serum, the activity remaining rate was measured as follows.
BTMA ₂ and each BTMA ₂ compound were dissolved in 50% methanol to 1280 μg / mL, and 0.05 mL of the solution and 0.45 mL of rat serum were mixed. When each sample and serum were mixed, the serum contact time was set to 0 minutes, and 0.1 mL was extracted from the mixture at 0 minutes, 15 minutes, 30 minutes, 60 minutes, and 120 minutes, and 0.3 mL of methanol was added to remove protein. went. After centrifugation at 12000 rpm for 5 minutes, 0.08 mL of the supernatant was infiltrated into an 8 mm thick paper disk.
The paper disk for preparing the calibration curve was prepared by adding 1.8 mL of DW (distilled water) to 0.2 mL of 1280 μg / mL 50% methanol solution of each BTMA ₂ compound to prepare a 128 μg / mL solution, and then performing two-stage dilution. Dilution series of 2, 4, 8, 16, 32, 64, and 128 μg / mL were prepared. 0.08 mL of these dilutions were soaked into each paper disk.
Place a paper disc soaked with BTMA ₂ and each BTMA ₂ compound on TSA (Tryptic Soy Agar) coated with 10 ⁸ cfu / mL of Enterococcus faecalis bacterial solution cultured overnight, and at 4 ° C for about 2 hours After pre-diffusion, the cells were cultured at 35 ° C. for 18-20 hours. After the culture, the diameter of the inhibition circle (fungal growth suppression range) was measured. A paper disk prepared for the calibration curve was also prepared in the same manner, and the concentration of BTMA ₂ and each BTMA ₂ compound was calculated from the calibration curve obtained from the diameter and concentration. The concentration of BTMA ₂ and each BTMA ₂ compound of serum contact time 0 minutes, based on serum contact time 0 min, 15 min, 30 min, 60 min, the percentage concentration of BTMA ₂ and each BTMA ₂ compound at 120 minutes The activity remaining rate was calculated.

Test example 3
A test was conducted in the same manner as in Test Example 2 except that BTMA ₂ shown in Table 3 and the BTMA ₂ compound of the present invention were used and methanol was not added for deproteinization. The results are shown in Table 3.

Test example 4
Except for using BTMA ₂ compound of the present invention shown in Table 4 and using Muller Hinton agar medium (MHA) coated with MRSA fungus instead of TSA coated with Enterococcus faecalis fungus, A test similar to Test Example 2 was performed. The results are shown in Table 4.

Test Example 5
A test was conducted in the same manner as in Reference Example 4 except that the BTMA ₂ compound of the present invention shown in Table 5 was used and methanol for protein removal was not added. The results are shown in Table 5.

From the results of Test Examples 2 to 5, it was found that the BTMA ₂ compound of the present invention did not lose its antibacterial activity even when administered into serum, and had very high metabolic stability.

The BTMA ₂ compound of the present invention exhibits excellent antibacterial activity and a broad antibacterial spectrum, and the antibacterial activity does not disappear even when administered orally or subcutaneously. Therefore, the antibacterial agent of the present invention containing the BTMA ₂ compound exhibits excellent antibacterial activity and a broad antibacterial spectrum, and has an excellent effect that the antibacterial activity does not disappear even when administered orally or subcutaneously. Furthermore, the BTMA ₂ compound of the present invention can be suitably used for the treatment of infections caused by MRSA.

  While this invention has been described in conjunction with its embodiments, we do not intend to limit our invention in any detail of the description unless otherwise specified and are contrary to the spirit and scope of the invention as set forth in the appended claims. I think it should be interpreted widely.

Claims (7)

A compound represented by the following general formula (1) or a salt thereof.

[Wherein, X represents a group represented by any one selected from the group consisting of the following general formulas (2) to (5).

In the general formula (2), R ²¹ represents a hydrogen atom, a hydroxy group, an alkyl group, an alkylthio group or an amino group.

In general formula (3), R ³¹ and R ³² each independently represent a hydrogen atom or an alkyl group, and may be bonded to each other to form a ring.

In the general formula (4), R ⁴¹ and R ⁴² each independently represent a hydrogen atom or an alkyl group.

In general formula (5), R ⁵¹ , R ⁵² and R ⁵³ are each independently a hydrogen atom, a hydroxy group, an alkyl group, a mercapto group, an alkylthio group, an amino group, an alkoxy group, a phosphate group or an aromatic heterocyclic group. And may be bonded to each other to form a ring. ] The group represented by the general formula (2) is a group represented by any one selected from the group consisting of the general formula (2-1), the general formula (2-2), and the general formula (2-3). The compound according to claim 1 or a salt thereof, wherein

[In General Formula (2-1), R ¹²¹ and R ¹²² each independently represent a hydrogen atom or an alkyl group, and may be bonded to each other to form a ring.

In General Formula (2-2), R ²²¹ represents a hydrogen atom, a hydroxy group, or an alkyl group.

In general formula (2-3), ^R321 represents an alkyl group. ] The group represented by general formula (5) is any one selected from the group consisting of general formula (5-1), general formula (5-2), general formula (5-3) and general formula (5-4) The compound or a salt thereof according to claim 1, which is a group represented by one.

[In General Formula (5-1), R ¹⁵¹ and R ¹⁵² each independently represent an alkyl group, and may be bonded to each other to form a ring. R ¹⁵³ represents a hydrogen atom or an alkyl group.

In general formula (5-2), ^R252 represents a hydrogen atom, an alkyl group, or an aromatic heterocyclic group.

In general formula (5-3), R ³⁵¹ and R ³⁵² each independently represent a hydrogen atom or an alkyl group, and may be bonded to each other to form a ring.

In general formula (5-4), ^R451 and ^R452 each independently represents an alkyl group, and may be bonded to each other to form a ring. ]   The antibacterial agent containing at least 1 sort (s) of the compound in any one of Claims 1-3, or its salt.   Use of the compound according to any one of claims 1 to 3 or a salt thereof as an antibacterial agent.   The compound or a salt thereof according to any one of claims 1 to 3, for use in the treatment of an infection caused by MRSA.   A method for treating an infection with MRSA, comprising administering the compound according to any one of claims 1 to 3 or a salt thereof.