JPH0354956B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a novel macrolide antibiotic. Various 16-membered ring macrolide antibiotics such as the leucomycin group, tylosin, and coelamycin are known. These exhibit strong antibacterial activity, especially against Gram-positive bacteria, but have a common drawback of relatively weak antibacterial activity against certain Gram-negative bacteria. Therefore, it has been desired to provide a 16-membered ring macrolide antibiotic that has strong antibacterial activity against both Gram-negative and Gram-positive bacteria and exhibits high blood concentrations. The present invention provides novel compounds derived from certain known 16-membered ring macrolide antibiotics that have strong antibacterial activity against both Gram-positive and Gram-negative bacteria and are easy to produce. It is based on this knowledge. Accordingly, an object of the present invention is to provide a 16-membered ring macrolide antibiotic that has improved antibacterial activity and can be easily produced. The present invention provides a compound represented by the following general formula () and a pharmacologically acceptable salt thereof. (In the formula, R represents a hydrogen atom or an acyl group consisting of C ₂ to _6. A is

[Formula] or C=O, B is a single Indicates a bond or an oxygen atom. X is

[Formula] (i) R ₁ and R ₂ each or either of them is _a hydrogen atom, a C _1-4 alkyl group, a C _3-10 cycloalkyl group, or -(CH _{2 )
o} −Ph (n=0, 1, 2). Ph is unsubstituted or phenyl substituted with an amino group, a _{C1-4 alkoxycarbonyl} group, a halogen, _{a C1-4} alkyl group, or _{a C1-4} alkoxy group. (ii) Either R ₁ or R ₂ is hydrogen or

[Formula] ₍ R ₃ is hydrogen or C _1-4 alkyl group). Or X is

【formula】

【formula】

【formula】

[Formula] and _a formula in which _the above heterocycle is substituted with a C _1-4 alkyl group, a C _1-4 alkoxy group, an amino group, or a halogen. ) The compound represented by formula () is a gram-positive bacterium,
It has antibacterial activity against Gram-negative bacteria, and has antibacterial activity against Gram-positive bacteria that is comparable to or slightly stronger than the antibiotic tylosin and demycarosyl tylosin derived from it. Also, especially
is a benzylamino group, 20-deoxo-20-
(N-benzylamino)-demycarosyltylosin Mycobacterium smegmatis ATCC
It showed the strongest antibacterial activity against 607 among tylosin, demycarosyl tylosin, and their related compounds. Furthermore, when administered orally in therapeutic experiments using mice infected with Streptococcus biogenes, this compound exhibited twice as strong antibacterial activity as the raw material demycarosyltylosin. In addition, the derivatives in which the benzylamino group was substituted with various amino groups, various halogen atoms, and alkyl groups were found to be comparable to or superior to the raw material demycarosyltylosin in the aforementioned infection treatment experiments in mice (in vivo). The results were shown. In addition, the aromatic alkylamino group represented by X in formula () includes anilino, N-methylanilino, and the like in addition to benzylamino. Examples of pharmacologically acceptable salts of the compound of formula () include salts with inorganic acids such as hydrochloric acid and phosphoric acid, and salts with organic acids such as acetic acid, propionic acid, citric acid, tartaric acid, and sulfonic acid. Can be mentioned. Examples of the physicochemical properties of the compounds according to the present invention are as follows. In formula (), R is a hydrogen atom, A is a carbonyl group, B is a single bond, and X is a benzylamino group (see Example 1) Elemental analysis, molecular formula and molecular weight Actual value C: 64.10 H: 8.60 N: 3.20
O: 24.10 Calculated value C: 64.04 H: 8.58 N: 3.25
O: 24.13 C ₄₆ H ₇₄ N ₂ O ₁₃ (Molecular weight 862) Melting point 88.5-91.5℃ Specific rotation [α] ²⁹ _D = -35.2° (c = 1, methanol) Ultraviolet absorption spectrum λ ^CH3OH _nax = 284 nm (ε, 22000) Infrared absorption spectrum 3440, 2950, 2980, 1710, 1680, 1590, 1460, 1350, 1165, 1090, cm ^-1 (KBr method) Mass spectrum (m/z) 756, 672, 565, 482, 174, 91 Thin layer chromatography Merck TLC plate, Kieselgel 60F _{254Developing} solvent: Chloroform/methanol/concentrated aqueous ammonia (10:1:0.05) Rf=0.20 Hydrogen nuclear magnetic resonance spectrum 90MHz nuclear magnetic resonance spectrum in deuterated chloroform is As shown in FIG. Note that tetramethylsilane was used as an internal standard. The acute toxicity (oral and intraperitoneal administration in mice) of the compounds according to the invention is approximately comparable to that of other 16-membered ring macrolide antibiotics. Tylosin is currently widely used as a veterinary drug, especially as a feed additive for the purpose of promoting growth. The antibacterial spectra of some of the compounds according to the invention are comparable or expanded or improved compared to the known 16-membered macrolide antibiotic tylosin and related compounds. Therefore, the compound according to the present invention is a new antibiotic and is expected to be used not only as a veterinary drug but also as a pharmaceutical. Table 1 shows an example of the antibacterial spectrum of the compound according to the present invention, and compares it with the raw material demycarosyltylosin, tylosin, and various amino derivatives of tylosin. In addition, the compound names in Table 1 are as follows. A: 20-deoxo-20-(N-dimethylamino)
Tylosin B: 20-deoxo-20-(benzylamino)-Tylosin C: 20-deoxo-20-anilinotylosin D: 20-deoxo-20-(para-ethoxycarbonylanilino)Tylosin E: 20-deoxo-20 -[N-(4-methylpiperazyl)amino]tylosin F: 20-deoxo-20-morpholinotylosin G: 20-deoxo-20-(N-methylanilino)
Tylosin H: 20-deoxo-20-(N-cyclohexylamino)demycarosyltylosin I: 20-deoxo-20-(N-benzylamino)-
Demycarosyltylosin J: 20-deoxo-20-(N-methylanilino)
Demycarosyltylosin K: 20-deoxo-20-morpholinodemycarosyltylosin L: Tylosin M: Demycarosyltylosin

【table】

[Table] As is clear from Table 1, compared with the known antibiotics tylosin and demycarosyl tylosin, the compounds [20-deoxo-20-(N-benzylamino) demycarosyl tylosin] and compound K
[20-deoxo-20-(N-morpholino)demycarosyltylosin] showed increased antibacterial activity against Mycobacterium smegmatis ATCC 607 and Escherichia coli NIHJ. Furthermore, in a treatment experiment using mice for Streptococcus biogenes infection, the ED ₅₀ of the compound when administered orally was 39 mg/Kg. This is the raw material compound M (demycarosyltylosin).
The therapeutic effect was approximately twice as good as that of ED ₅₀ 76mg/Kg. Other derivatives are also expected to have therapeutic effects. Examples of methods for producing compounds according to the present invention are as follows. General formula () with tylosin as the representative compound (In the formula, A, B, and R have the same meanings as in the formula ()) as a starting material, and by increasing the concentration of the acid using an inorganic acid such as hydrochloric acid or sulfuric acid. , or by increasing the reaction time or increasing the heating temperature, the mycarose moiety is eliminated, and the general formula () (In the formula, A, B and R have the same meaning as the above formula ()) [US Patent 3433711: Tetrahedron Lett.No.34.2339]
(1964): No. 40.4737 (1970)]. These 16-membered ring lactone compounds represented by the general formula () are dissolved in methanol. Next, ammonia, primary amine, and secondary amine are used as amination reagents. While stirring the reaction solution in a nitrogen stream or air, sodium cyanoborohydride or lithium cyanoborohydride dissolved in methanol is added, and the mixture is reacted at room temperature under neutral conditions (usually pH 6 to 8). This reaction usually proceeds at room temperature, but the reaction can be controlled by heating or cooling if necessary. The reaction solution obtained as above was poured into cold sodium bicarbonate water, extracted with an organic solvent such as benzene, ethyl acetate, chloroform, etc. under weak alkaline conditions, and the organic solvent layer was collected and concentrated. Thus, a coarse powder of the compound represented by the general formula () is obtained. Furthermore, the compound represented by the general formula () can also be produced by the following method, which is different from the above-mentioned method. That is, an amino derivative is obtained by once reacting the compound represented by the general formula () with an aminating reagent by the above method. Next, this amino derivative is subjected to an acid hydrolysis reaction using an inorganic acid such as hydrochloric acid or sulfuric acid to remove the mycarose moiety, thereby obtaining a crude powder of the compound represented by the general formula (). The crude powder is purified by known methods such as a transfer method using an organic solvent and column chromatography using an adsorbent such as silica gel or alumina. By acylating the aminated macrolide compound thus obtained by a conventional method, a compound represented by the formula () can be obtained. That is, in the presence of an acylation reaction solvent for an aminated macrolide compound, such as pyridine, triethylluamine, p-toluenesulfonic acid, etc., a carboxylic acid anhydride or acid halide having 2 to 5 carbon atoms is used as an acylating agent. Although the reaction is generally carried out at room temperature, it may be carried out under ice cooling or heating if necessary. Acylated reaction product obtained in this way ()
Methods include recrystallizing using a recrystallization solvent, or performing column chromatography using alumina, silica gel, etc., separating only the target substance fraction, and precipitating crystals by operations such as concentration. Purify by If desired, a pharmaceutically acceptable compound can be prepared from the compound of formula () by a conventional method. Example 1 20-Deoxo-20-(N-benzylamino)demycarosyltylosin Tylosin (5 g) was mixed with benzylamine (7.9 g)
and sodium cyanoborohydride (1.4 g) were dissolved in methanol (50 ml) and reacted at room temperature in a nitrogen stream. After the reaction was completed, the reaction solution was poured into cold saturated sodium bicarbonate water, and chloroform (100ml
(twice). The extract was concentrated, and the resulting crude powder was purified by silica gel column chromatography [developing solvent: chloroform/methanol/concentrated aqueous ammonia (15:1:0.05)] to obtain an amino derivative, 20-deoxo-20-( N-benzylamino)
Tylosin (3.8g) was obtained. This material (1 g) was dissolved in 0.1N hydrochloric acid (30 ml) and reacted at room temperature for 19 hours. After the reaction is complete, wash the reaction solution with chloroform (30ml) and remove the remaining aqueous layer.
After adjusting the pH to 8.0 with 1N sodium hydroxide solution,
Extracted with chloroform (3x100ml). The chloroform layer was dried over anhydrous sodium sulfate and then concentrated to dryness under reduced pressure to obtain a pale yellow crude powder (820 mg).
This powder was purified by silica gel column chromatography [developing solvent: chloroform/methanol/concentrated aqueous ammonia (20:1:0.05)] to obtain a white powder (620 mg) in a yield of 72%. The physicochemical properties of this substance were as described above. Example 2 20-deoxo-20-(N-methylamino)demycarosyltylosin 20-deoxo-20-(N- Methylamino)tylosin (300 mg) was dissolved in 0.2N hydrochloric acid and left at room temperature for 3 hours. After the reaction was completed, the target substance was isolated from the reaction solution by the method described above. Melting point 95.5-98.5℃ Ultraviolet absorption spectrum λ ^MeOH _nax = 283 nm (ε, 20600) Mass spectrum (m/z) 862, 688, 190, 174 Hydrogen nuclear magnetic resonance spectrum δ 1.77 (s, H ₂₂ ), 2.49 (s , C ₃ , −N
(Me) ₂ ), 2.87 (s, C ₂₀ −N−Me), 3.46 (s, C ₂ ″−
OMe), 3.59 (s, C ₃ ″, −OMe), 4.33 (d, H ₁ ), 4.56 (d, H ₁ ″, 5.00 (dt, H ₁₅ ), 5.84 (bd,
H ₁₃ ), 6.63 (d, H ₁₀ ), 6.70 (d, (ortho-H)-anilino), 6.73 (d, para-H-anilino), 7.20 (d,
(meta-H)-anilino), 7.2 (H ₁₁ ) Yield 43% Example 3 20-deoxo-20-(N,N-dibenzylamino)demicalosyltylosin Tylosin (1 g) was mixed with 0.2N hydrochloric acid (30 ml). ) and reacted at room temperature for 4 hours. After the reaction was completed, the reaction solution was washed with chloroform (20 ml), the remaining aqueous layer was adjusted to pH 8.0 with 1N sodium hydroxide solution, the chloroform layer was dried over anhydrous sodium sulfate, and then concentrated to dryness under reduced pressure. A pale yellow powder (820 mg) was obtained. This coarse powder, dibenzylamine (0.5ml)
and sodium cyanoborohydride (330 mg) were dissolved in methanol (10 ml) and reacted in a nitrogen stream at room temperature for 4 hours. After the reaction was completed, the reaction product was poured into ice water, and saturated aqueous sodium hydrogen carbonate solution (50%
ml) and extracted with chloroform (150 ml). The extract was dried over anhydrous sodium sulfate and then concentrated to dryness under reduced pressure to obtain a crude substance. This powder was subjected to silica gel column chromatography [Developing solvent: chloroform/methanol/concentrated ammonia water (25:
1:0.05)] to obtain the target substance (950 mg).
Got it at 47%. Specific optical rotation [α] ²⁹ _D = -29.4° (c = 1, methanol) Mass spectrum (m/z) 952, 726, 175 Hydrogen nuclear magnetic resonance spectrum δ 1.77 (s, H ₂₂ ), 2.46 (s, C ₃ , -N
(Me) ₂ , 3.50 (s, C ₂ ″−OMe), 3.60 (s, C ₃ ″−
OMe), 4.54 (d, H ₁ ″), 5.06 (d, H ₁₅ ), 5.90 (bd,
H ₁₃ ), 6.25 (d, H ₁₀ ), 7.3 (benzyl-H) Example 4 20-deoxo-20-(N-cyclohexylamino) demycarosyl tylosin Demycarosyl tylosin (500 mg), cyclohexylamine (450 mg) and sodium cyanoborohydride (120 mg).
After the reaction was completed, the target substance was obtained from the reaction solution by the method described above. Melting point 98.5-103.0℃ Specific rotation [α] ²⁹ _D = -21.9゜ (c = 1, methanol) Ultraviolet absorption spectrum λ ^MeOH _nax = 284nm (ε, 22400) Mass spectrum (m/z) 854, 664, 473 , 175, 174 Yield 93% Example 5 20-deoxo-20-(N-morpholino) demycarosyltylosin Using demycarosyltylosin (500 mg), morpholine (0.6 ml) and sodium cyanoborohydride (160 mg) , carried out the reaction. After the reaction is complete,
The target substance from the reaction solution was isolated by the method described above. Melting point 108.0-110.0℃ Ultraviolet absorption spectrum λ ^MeOH _nax = 283 nm (ε, 19500) Mass spectrum (m/z) 842, 824, 669, 477, 461, 174 Yield 44%

[Brief explanation of drawings]

FIG. 1 shows the nuclear magnetic resonance spectrum of the material according to Example 1.

Claims (1)

[Claims] 1 General formula () (In the formula, R represents a hydrogen atom or an acyl group consisting of _C2-6 . A represents [formula] or C=O, and B represents a single bond or an oxygen atom. X is represented by [formula] , (i) R ₁ and R ₂ each or either of them is a hydrogen atom, a C _1-4 alkyl group, _{a C 3-10 cycloalkyl} group, or -(CH ₂ )
_o −Ph (n=0, 1, 2). Ph is unsubstituted or phenyl substituted with an amino group, a _{C1-4 alkoxycarbonyl} group, a halogen, _{a C1-4} alkyl group, or _{a C1-4} alkoxy group. (ii) Either R ₁ or R ₂ is hydrogen or represented by the formula (R ₃ is hydrogen or _{a C 1-4 alkyl} group). Alternatively _, X is represented by [Formula] [Formula _] [Formula] [Formula] and a formula in which the above heterocycle is substituted with a C _1-4 alkyl group, a C _1-4 alkoxy group, an amino group or a halogen. ) or a pharmacologically acceptable salt thereof. 2 R is a hydrogen atom, an acetyl group, a propionyl group, a butyryl group, an isobutyryl group, or an isovaleryl group, and R are the same or different,
A compound according to claim 1 or a pharmacologically acceptable salt thereof.