KR830002482B1 - 3" - acylated macrolide antibiotics - Google Patents

Abstract

Macrolide antibiotics I (R3=C2˜C6 alkanoy1; R4=C2˜C5 alkanoy1; R11=C2˜C3 alkanoy1) were ! prepd. Thus, 2'-o-acetylleucomycin A5, dissolved in dichloro methane, was acetylated to the 2'-c-acety1-3,9-di-o-chloroacetylleucomycin A5, which was treated with NH3-MeOH to give 2'-o-acetyl-3"-o-isovalerylleucomycin A5. The resulting material was dissoloved in MeOH, refluxed for 20 hrs., and dired to give 3"-o-isovalerylleuconycin A5.

Description

Method for preparing 3 "-acylated macrolides

The present invention relates to a method for preparing a new 3 "-acylated macrolide antibiotic having the following general formula (I).

Wherein R ₃ is an alkanoyl group (C ₂ -C ₆ ),

R ₄ is an alkanoyl group (C ₂ -C ₅ ).

Also useful are salts of the compounds, where salt means a physiologically acceptable salt. Examples of such salts are inorganic salts such as hydrogen chloride, sulfate or phosphate and organic salts such as acetite, propionate, tartrate, citrate, succinate, aspartate or glutamate. Other nontoxic salts may also be included.

The novel compounds (1) of the present invention are conventional 16-membered macrolides such as leucommycin group antibiotics, including irradiation mycin, SF-837 group antibiotics, YL-704 group antibiotics, and Espinomycin group antibiotics. It has improved antimicrobial activity against susceptible or resistant strains compared to antibiotics.

In particular, the compounds are very effective against strains that are resistant to other macrolide antibiotics such as onremycin, erythromycin, carbomycin and spiramycin. Furthermore, a series of blood levels increases because deacylation at the 4 "position, which is one of the reasons for inactivation of 16-membered macrolide antibiotics, cannot easily occur. Syrups for infants with difficult pill administration may be advantageously prepared because of their strong properties and reduced persistent bitterness, and as a result, the compound (1) of the present invention is expected to exhibit an excellent clinical infectious disease treatment effect.

In the nomenclature of the compounds of the invention, this is influenced by substituents at the 3-position of the general formula (1) or substituents at the 3 "and 4" positions. Thus, when the 3 "position is acylated or when the original acyl group at the 4" position is not translocated to the 3 "position as an acyl arrangement, the nomenclature is a known antibiotic represented by the following structural formula (2) Based on that.

Wherein R ₁ is a hydrogen atom or an alkanoyl (C ₂ -C ₃ ) group,

R ₄ is an alkanoyl (C ₂ -C ₅ ) group.

The compounds represented by the above formula are listed as follows.

The antibiotic of structure (2) wherein R ₁ is a hydrogen atom has a hydroxyl group at the 3,9,2'- and 3 "positions, and when the R ₁ is an acetyl or propionyl group, It has a hydroxyl group.

In these groups, hydroxyl groups at the 3, 9 and 2 'positions are easily acylated and therefore many acylated derivatives therefrom have been reported. However, hydroxyl groups at the 3 "position have been reported to be inactive.

In recent years, derivatives in which hydroxyl groups at the 3 "position are acylated have been reported. (See Japanese Patent Publication Nos. 49-124087 and 51-26887.) The reported compounds are essentially propionyl and 9 positions. To have acyl groups.

However, in order to produce derivatives (hereinafter referred to as 3 "-acylated derivatives) having 3" -acylated compounds having at least one hydroxyl group at 3 and 9 positions, the acyl group is introduced only at the 3 "-position of the antibiotics previously known. In order to achieve this, acylation by the conventional acylation method was impossible because there is a very active hydroxyl group at a position other than the 3 ″ position.

We protect by hydroxyl groups other than 3 ", in particular by protecting groups which can be easily removed without 3" deacylation after the 3 "and 9 position hydroxyl groups are acylated. I figured it could be.

Accordingly, an object of the present invention is to provide a method for producing a novel macrolide antibiotic represented by the general formula (1).

Another object of the present invention is to provide an antibiotic having high blood levels when taken and at the same time showing high viability against sensitive or resistant strains.

Another object of the present invention is to provide a novel antibiotic with a strong, persistent bitter taste.

The novel 3 "-acylated macrolide antibiotics of general formula (I) according to the invention are prepared as follows.

Where R ₄ is as defined above,

R ₃ is an alkanoyl group (C ₂ -C ₄ ).

The 3 'and 9'-position hydroxyl groups of the 2'-acylated antibiotic of the general formula (3) were treated with acetyl halide chloride in an inert organic solvent in the presence of a tertiary organic amine to prepare a compound of the following general formula (4). do.

Wherein R ₄ and R ₈ are as defined above,

R ₅₁ is an acetyl chloride group.

This compound (4) was treated with an aliphatic carboxylic acid (C ₂ -C ₆ ) halide in an inert organic solvent in the presence of tertiary organic amine, 3 "-acylated to form a compound of the general formula (5), followed by methanol or Treatment with ammonia in ethanol affords the compound of formula (6).

Wherein R ₃ , R ₄ , R ₈ and R ₅₁ are as previously defined.

The compound (6) is heated in methanol, which may contain any water, to remove the acyl group at the 2 'position, resulting in the 3' '-acylated macrolide antibiotic of general formula (1) as the final desired product. .

The compound of formula (4) is a 2'-acylated antibiotic (3) which introduces a protecting group to prevent acylation of the 3 and 9 position hydroxyl groups during the subsequent 3 "-acylation reaction. The hydroxyl group is also protected by another protecting group, with the preferred protecting group being an ethethyl group.

The hydroxyl groups at positions 3 and 9 are protected by acetyl chloride groups such as chloroacetyl, dichloro acetyl or trichloro acetyl groups. Introduction of the protecting group is carried out with 2-3 fold excess of acetyl chloride halide. The 2'-acylated antibiotic 10 described above is described in Japanese Patent Publication No. 53-7434 and J. Med. Chem. 2'-acylated derivatives of antibiotics (2) wherein R ₁ is a hydrogen atom, such as leucomycin A ₁ , A ₅ , A ₇ or A ₉ specified in 20 (5), 732-736 (1977) it means.

Compound (4) is 3 "-acylated by aliphatic carboxylic acid halide. The reaction is carried out by heating in an inert organic medium in the presence of a tertiary organic amine. Preferred inert organic solvents are acetone, methyl ethyl ketone, ethyl acetate , Dimethoxyethane, tetrahydrofuran, dioxane, benzene or toluene, and the like, and tertiary organic amines include pyridine, piclin or collidine.

In addition, triethylamine, dimethylaniline, tribenzylamine, N-methylpiperidine, N-methylmorpholine, quinoline or isoquinoline can also be used as the tertiary organic amine. Preferred aliphatic carboxylic acid halides are aliphatic carboxylic acid (C ₁ -C ₆ ) halides such as acetylchloride, propionyl chloride, butyryl chloride, isobutyryl chloride, isovaleryl chloride or caproyl chloride. The reaction heating temperature is about 50-120 ° C., and the reaction time may vary depending on the reaction temperature. The progress of the reaction can be detected by silica gel thin film chromatography, and the reaction termination is usually determined within 1-150 hours.

Compound (5) produced by the reaction as described above can be separated by adjusting the pH of the reaction mixture to 8-10 and precipitated in water. On the other hand, when the reaction solvent is an organic solvent that does not stand in water, the reaction mixture is poured into water, the pH is adjusted to 8-10 and then extracted together with an organic solvent that does not stand in water.

Purification can be carried out by silica gel, activated alumina or adsorbent resin chromatography using benzene-acetone or the like as the eluent. Removal of the protecting group at position 3 or 9 can be done by treating compound (5) with ammonia in methanol in room temperature. The termination of the reaction can be checked by disappearing the point of compound (5) on the silica gel thin film chromatogram.

Compound (6) obtained by distilling ammonia and methanol off from the reaction mixture removes 2'-acyl groups by heating in methanol, which may contain water. The final product (1) is obtained by separating and purifying the compound obtained by removing methanol.

Separation of compound (1) from the reaction mixture is well known and well known separation and purification methods such as concentration, extraction, washing, mobile extraction and recrystallization, chromatography using silica gel, activated alumina, adsorbent resin or ion exchange resin. It is done according to

The antimicrobial spectra of compound (1) of the present invention is shown in the following table.

These data show that the compounds have improved antimacrobial spectractivity for sensitive and resistant strains when compared to existing antibiotics.

Minimum Inhibition Concentration (MIC)

ug / ml

/ml의 에리스로마이신, 올레엔도 마이신, 16-멤버 매크롤라이드항체, broth 희석법* Clinically isolated (macrolide antibody A-group strain) inoclum size 10

/ ml erythromycin, oleendomycin, 16-membered macrolide antibody, broth dilution

** LM = leucomycin

The present invention will be described in detail with reference to the following Examples. Unless otherwise indicated, the Rf value is measured by the following thin film chromatography.

Media: Silica gel 60 (Art. 5721, Merck Co.)

Developer:

A: n-hexane-benzene-acetone-ethyl acetate-methanol (90: 80: 25: 60: 30)

B: benzene-acetone (3: 1)

C: benzene-acetone (5: 1)

Example 1

3 "-0-Isovaleryl Leucomycin A ₅ :

Dry dicicle was added to dry pyridine (1.64 ml) in 2'-0-acetyl leucomycin A ₅ (5 g) dissolved in methane (25 ml), followed by dropwise addition of dichloro acetyl chloride (1.77 ml) under ice cooling for 1 hour. Stir while. Cold water is added to the reaction mixture and extracted with dichloromethane (25 ml).

The dichloro methane layer was dehydrated with anhydrous sodium sulfate and dried in vacuo to yield 2'-0-acetyl-3,9-di-0-chloroacetylleucoacin A ₅ quantitatively.

This product (5 g) was dissolved in dry dioxane (50 ml), r-clidine (7.87 ml) was added, and isovaleryl chloride (6.62 ml) was added dropwise under cooling, followed by stirring at 90 ° C. for 80 hours. The reaction mixture is poured into cold water (500 ml) and extracted twice with benzene (400 ml). The benzene layer is dehydrated with anhydrous magnesium sulfate and dried in vacuo.

The residue is dissolved in methanol (20 ml), ammonia saturated methanol solution (20 ml) is added and stirred at room temperature for 30 minutes.

The reaction mixture is poured into cold water (500 ml), extracted twice with benzene (400 ml), dehydrated with anhydrous magnesium sulfate and dried in vacuo. The residue is chromatographed on silica gel tubes using benzene-acetone (15 :: 1, 7: 1 and 5: 1).

The main parts were collected and dried in vacuo to give 2'-0-acetyl-3 "-0-isovalerylleucomycin A ₅ (1.255), which was dissolved in methanol (20 ml) and heated to reflux for 20 hours. . and dried in vacuo the residue was benzene-acetone (9: 1) was used to collect the portion is curled, representing by chromatography on a grafted silica gel tube Rf _a = 0.48 is obtained object product (830mg) Rf = 0.48 _a

Mass (m / e): 855 (M ⁺ ) 768 (M ⁺ -87) 754 (M ⁺ -101)

NMR (CDCl ₃ , 100 MHz: 1.42 (3'-CH ₃ ), 9.89 (CHO) ppm

Example 2

3 "-0-acetylleucomycin A ₁ :

Dry pyridine (1.64 ml) was added to 2'-0-acetylleucomycin A ₁ (5 g) dissolved in dry dichloromethane (25 ml), followed by dropwise addition of dichloro acetyl chloride (1.77 ml) under ice cooling for 1 hour. Stir. Ice water (25 ml) is added to the reaction mixture, which is extracted with dichloromethane (25 ml). The dichloro methane layer was dehydrated with anhydrous magnesium sulfate and dried in vacuo to yield 2'-0-acetyl-3,9-di-0-dichloro acetylleucomycin A ₁ quantitatively. To this product (5 g) dissolved in dry ethyl acetate (50 ml) r-collidine (9.4 ml) and acetyl chloride (4.6 ml) were added under ice cooling and stirred at 70 ° C. for 72 hours.

The reaction mixture is poured into cold water (250 ml) and extracted twice with chloroform (150 ml). The chloroform layer is washed with dilute hydrochloric acid (pH 2), water, saturated aqueous sodium carbonate, and then dehydrated with anhydrous sodium sulfate and dried in vacuo.

Dissolve the residue in methanol (10 ml), add ammonia saturated methanol solution (10 ml) and stir at room temperature for 30 minutes. The reaction mixture is poured into cold water (250 ml) and the chloroform is extracted twice. The chloroform layer is dehydrated with anhydrous magnesium sulfate and dried in vacuo.

The residue was chromatographed on a silica gel tube using benzene-acetone (8: 1) and the main part was concentrated to give a coarse powder of 2 ', 3 "-di-0-acetylleucomycin A ₁ (1.4 g).

This powder is dissolved in methanol (20 ml), heated to reflux for 20 hours and dried in vacuo. The residue is chromatographed on a silica gel tube with benzene-acetone (6: 1) to collect and dry the portion where Rf _A = 0.46 to give the desired product (716 mg). Rf _A = 0.46

Mass (m / e): 827 (M ⁺ ), 768 (M ⁺ -59), 726 (M ⁺ -101)

NMR (CDCI ₃ , 100 MHz): 1.43 (3 ″ -CH ₃ ), 2.00 (3 ″ -OAc), 9.89 (CHO) ppm.

Example 3

3 "-0-propionyl leucomycin A ₅

In Example 1, 2'-0-acetyl-3 "-0-propionyl leucomycin A ₅ is prepared using propionyl chloride (4.72 ml) instead of isovaleryl chloride (6.62 ml).

This compound is dissolved in methanol (20 ml), refluxed for 16 hours and dried in vacuo. The residue is dissolved in a small amount of benzene and chromatographed on a silica gel tube with benzene-acetone (6: 1). Collect and dry the fractions showing Rf _A = 0.47 to give the desired product (850 ml).

Rf _A = 0.47, Rf _B = 0.14

Mass (m / e): 827 (M ⁺ ), 754 (M ⁺ -73), 740 (M ⁺ -87)

NMR (CDCI ₃ , 100 MHz): 1.44 (3 ″ -CH ₃ ), 2.57 (3′-N (CH ₃ ) ₂ ), 3.56 (4-OCH ₃ ), 9.86 (CHO) ppm.

Example 4

3 "-0-acetyl leucomycin A ₅

In Example 2, 2 ', 3 "-di-0-acetyl leucomycin A ₅ (1.4 using 2'-0-acetyl leucomycin A ₅ instead of 2'-0-acetyl leucomycin A ₁₎ was used. g) is prepared.

The compound is dissolved in methanol (50 ml), refluxed for 15 hours and dried in vacuo. The residue is chromatographed on a silica gel tube using benzene-acetone (3: 1).

The fractions representing Rf _A = 0.45 were collected and vacuum dried to yield the desired product (710 mg).

Rf _A = 0.45, Rf _B = 0.10

Mass (m / e): 813 (M ⁺ ), 754 (M ⁺ -59), 726 (M ⁺ -87)

NMR (CDCl ₃ , 100 MHz): 1.43 (3 ″ -CH ₃ ), 2.02 (3 ″ -OAc), 9.88 (CHO) ppm.

Claims (1)

Compound (4) produced by treating and protecting hydroxyl groups at the 3 'and 9'positions of the 2'-acylated antibiotics of the general formula (3) with acetyl chloride halide in an inert organic solvent in the presence of tertiary organic amine In the presence of an inert organic solvent in the presence of an aliphatic carboxylic acid (C ₂ -C ₆ ) halide to 3 "-acylation to prepare a compound of formula (5), which is treated with ammonia in Forming the compound of 6) and then heating in a methanol, which may include any water to remove the acyl group in the 2 'position to prepare a 3 "-acylated macro of the general formula (1) for the ride antibiotic.

Wherein R ₃ and R ₄ are an alkanoyl group (C ₂ -C ₆ ) and an alkanoyl group (C ₂ -C ₅ ), respectively.

Wherein R ₄ is as defined above and R ₈ is an alkanoyl group (C ₂ -C ₄ ).

Wherein R ₄ and R ₈ are as defined above and R ₅₁ is an acetyl chloride group.

Wherein R ₃ , R ₄ , R ₈ and R ₅₁ are as previously defined.

Wherein R ₃ , R ₄ , and R ₈ are as previously defined.