KR19990014843A - Antibiotic - Google Patents

Abstract

The present invention relates to an N-alkyl-N-glycosyl derivative of an alkyl ester of an antimicrobial antibiotic of the polyene macrolide group in formula (1) (wherein M denotes the residue of a polyenemedrolide antibiotic , R represents a substituted sugar residue, and R ¹ and R ² denote a C _1-4 alkyl group) and a salt thereof (wherein R, R ¹ , R ² , And A represents an organic or inorganic acid anion). The compounds of formulas (a) and (b) in which the N-alkyl and alkyl ester groups are represented by methyl are particularly interesting.

The present invention also relates to methods of making the compounds, compositions containing them and their use in therapy, and the treatment of exogenous and endogenous homologous infections in humans and animals.

Description

Antibiotic

The present invention relates to an N-alkyl-N-glycosyl derivative of an alkyl ester of an antimicrobial antibiotic of the polyene macrolide group of the formula (1) (a), a salt of the compound 1 (a) The present invention relates to a process for preparing a compound of formula (1) (a) and (a) and a process for their use in medicine, wherein M in formula (1) , R is a substituent of the sugar residue, R ¹ and R ² may be the same or different and represent a C _1-4 alkyl group, and A means an anion of an inorganic acid or an organic acid.

Particularly interesting compounds are the N-methyl-N-glycosyl derivatives of the methyl esters of the antimicrobial antibiotics of the polyenmarmrolide group of formula (1) (b), their salts in formula (b) Wherein M is a polyenemedrolide residue, R is a substituent of a sugar residue, and A is an anion of an inorganic acid or an organic acid. do.

N-alkyl derivatives of antibiotics of the polyene macrolide group wherein the amino group of the parent antibiotic is substituted by an alkyl group are known.

N-glycosyl derivatives of polyenemaprolides in which the amino groups of the parent antibiotic are substituted with sugar residues are also described in J. Antibiotics 28 , 244 (1975), L. Falkowski, J. Gorik, P. Kolodziejczyk, J. et al. Pawlak, J. Zielinski, T. Ziminski, E. Burowski; Acta Polon. Pharm. 37 , 517 (1980), L. Falkowski, J. Pawlak, J. Gorik, P. Kolodziejczyk, B. Stefanska, E.Bylec, E, Borowski. Examples of sugars used in the preparation of these derivatives include D-glucose, D-mannose, L-rhamnose, D-ribose and maltose. When polyene macrolides and suitable sugars are reacted, perhaps an Amadori potential reaction occurs simultaneously to provide the corresponding N-glycosyl derivatives. Compounds have the advantage of exhibiting biological activity similar to the activity of the starting antibiotic and forming a water-soluble salt. However, the high toxicity associated therewith presents significant drawbacks.

J. Antibiotics 28 , 244 (1975), L. Falkowski, J. Golik, P. Kolodziejczyk, J. Pawlak, J. Zielinski, T. Ziminski, E. Bouowski; Acta Polon. Pharm. 37 , 517 (1980), L. Falowaski, J. Pawlak, J. Golik, P. Koldziejczyk, B. Stefanska, E.Bylec, E.Borowski], the amino acid of the parent antibiotic is D- glucose, D-mannose , L-rhamnose, D-ribose, or maltose are known. The compound is prepared by the reaction of polyenemedrolide with the sugar described above and the simultaneous potentiation reaction. The compounds exhibit biological activities similar to those of the starting antibiotic and form a water-soluble salt. However, they exhibit high toxicity.

Another type of derivative is known from J. Antibiotics 32 , 1080 (1979), L. Falkowski, B. Stefanska, J. Zielinski, E.Bylec, J. Gallik, P. Colodziejczyk, , Trimethylammonium derivatives of polyenemalcolide methyl ester, in which the amino group of the parent antibiotic is completely methylated to provide a quaternary ammonium salt. These compounds are prepared by fully methylating the parent antibiotic to dimethyl sulfate. The advantageous properties of the derivatives include their water solubility and antimicrobial activity similar to the starting antibiotic. Unfortunately, they are very toxic and unstable.

Another form of the derivative is the poly (2-hydroxyethyl) derivative known in J. Antibiotics 32 , 1080 (1979), L. Falkowski, B. Stefanska, J. Zielinski, E.Bylec, J. Gallik, P. Kolodziejczyk, And trimethylammonium derivatives of ene-dichloride methyl ester, in which the antibiotic methyl ester amino group is fully methylated to provide a quaternary ammonium salt. The compound is prepared by fully methylating the parent antibiotic to dimethyl sulfate. Derivatives are soluble in water and are characterized by antimicrobial activity similar to the activity of the starting antibiotic, but they are highly toxic and unstable.

Other forms of N-alkyl derivatives include antibiotics and micah-substituted maleimides such as N-ethylmaleimide, N, N'-hexamethylene dimaleimide, N- (3-dimethylaminopropyl) Succinimidyl derivatives formed by an addition reaction, and these compounds can be prepared according to the method described in J. Antibiotics, 44 , 979 (1991), A. Czerwinski, WAKonig, T. Zieniawa, P. Sowinski, Milewski, E.Borowski]. These compounds are less toxic than endogenous antibiotics, but their antibacterial activity is reduced.

Finally, the most recently known group of N-alkyl derivatives of polyenemapololide is the N-enamines formed by the reaction of antibiotics with antibiotics and acetylacetone, ethylacetyl acetate, dimethylacetal or dimethylformamide and Amidine derivatives, which are described in Acta Polonica Phann. 45 , 71 (1988), B. Stefanska, J. Zielinski, E.Borowski, L. Falkwski. These derivatives exhibit antibacterial activity similar to the activity of parent antibiotics and improve solubility in organic solvents, but still exhibit significant toxicity and are very unstable.

The present inventors have now produced N-alkyl-N-glycosyl derivatives of alkyl esters of mixed polyene macrolide antibiotics. Methods for the preparation of these mixed compounds have also been achieved. These novel compounds have a high antimicrobial activity similar to that of the parent antibiotic, forming a water-soluble acid addition salt, and the toxicity has been found to be significantly reduced. Because these compounds exhibit high toxicity, which is a significant drawback, N-alkyl derivatives of all the polyene macrolides of the prior art are not expected. The compounds included in the present invention have no such drawbacks.

To date, N-methyl-N-glycosyl derivatives of methyl esters of polyenemmacrolide antibiotics and methods for their preparation have not been known. Surprisingly, these compounds retained a high antimicrobial activity similar to that of the parent antibiotics, which formed water-soluble acid addition salts and markedly reduced toxicity. These compounds do not exhibit the same toxicity as the N-alkyl derivatives of the prior art. High toxicity is a fundamental drawback of all of the already known N-alkyl derivatives of polyene macrolides, and the compounds included in the present invention have no such drawbacks.

A first aspect of the present invention provides an N-alkyl-N-glycosyl alkyl ester of an antimicrobial of the macrolide group of formula (I).

In the above Formula 1 (a)

M means the residue of an antibiotic of the polyene macrolide group,

R means a part of the sugar residue formed by the reaction of a mono or oligosaccharide with an antibiotic,

R ¹ and R ² are the same or different and each represents a C _1-4 alkyl group.

Preferably, the antimicrobial residue of the polyene macrolide group M is selected from the group consisting of amphotericin B, candidin, candidoin, candidinin, mycoheptin, nystatin, polyfungin, aureofacin, vacidin, trichomycin, or candicidin.

The mono or oligosaccharide from which the sugar residue R is derived is preferably selected from D-glucose, L-glucose, D-mannose, D-galactose, lactose or maltose. Presumably, when the reversion reaction occurs at the same time, a suitable sugar reacts with the polyenemedrolide to provide an N-glycosyl precursor for the compound of the present invention.

A preferred embodiment of the present invention comprises an N-methyl-N-glycosyl derivative of a methyl ester of an antibiotic of a polyene macrolide group represented by the following formula (b): <

In the above formula (1) (b)

M represents a residue of an antibiotic substance of a polyene macrolide group,

R is a portion of the sugar moiety formed by the reaction of mono oligosaccharides, preferably D-glucose, L-glucose, D-mannose, D-galactose, lactose, or maltose with antibiotics and concurrent amaryl reactions.

In a preferred form, the present invention provides a method for treating or preventing an antibiotic substance in a polyenmarmrolide group, wherein the antibiotic substance of the polyenmarmrolide group is selected from the group consisting of amphotericin B, Candidin, Candidone, Candidinin, Maikoheptin, Nistatin, Polypuggin, Oreofacin, Bacidine, Lt; / RTI > derivatives.

A second aspect of the present invention provides a salt of an N-alkyl-N-glycosyl derivative of an antibiotic of a polyenmarmrolide group in formula (2)

In the formula (2)

M, R, R ¹ and R ² are as defined in the first aspect of the present invention,

A means an anion of an organic or inorganic acid.

The salt is preferably a physiologically acceptable salt, and a compound wherein A is an anion of L-aspartic acid is particularly preferred. Salts wherein R < ₁ > and R < ₂ > are methyl groups are particularly preferred.

A preferred embodiment of the second aspect of the present invention comprises a salt of an N-methyl-N-glycosyl derivative of an antibiotic of the polyenmarmrolide group represented by formula (b)

In the above formula (2)

M means the residue of an antibiotic of the polyene macrolide group,

R is part of the sugar moiety formed by the reaction of a mono or oligosaccharide, preferably D-glucose, L-glucose, D-mannose, D-galactose, lactox, or maltose with an antibiotic, ,

A is an anion of an organic or inorganic acid.

In a preferred form, the present invention provides a method for treating or preventing an antibiotic substance in a polyenmarmrolide group, wherein the antibiotic substance of the polyenmarmrolide group is selected from the group consisting of amphotericin B, Candidin, Candidone, Candidinin, Maikoheptin, Nistatin, Polypuggin, Oreofacin, Bacidine, Preferably a salt in which A is related to an anion of L-aspartic acid.

A third aspect of the present invention is a process for preparing a compound of formula (a) wherein M, R, R ¹ and R ² are as defined in the first and second aspects of the present invention, Reacting a polyene antibiotic with a mono or oligosaccharide to produce an N-glycosyl derivative of a polyenemarcholide antibiotic, characterized by the occurrence of a potential reaction; Possibly separating the product of the rearrangement reaction; Treating the product with an alkylating agent and purifying the crude product.

In a first embodiment of the third aspect of the present invention, the product of the potentiostatic reaction is separated in the form of a suspension by precipitation from a solution in which the potential reaction takes place. An organic solvent such as N, N-dimethylformamide is preferable for supporting the potentiostatic reaction. An organic solvent such as diethyl ether is suitable for carrying out the formation of a suspension by precipitation of the product of the potentiostatic reaction.

In a second embodiment of the third aspect of the present invention, the alkylation of the product of the potentially rearrangement reaction is carried out at a reduced temperature. A temperature of -5 ° C to + 5 ° C is preferred. Alkylating agents such as diazoalkanes may be used. The ethereal solution of the diazomethane is preferably an alkylating agent.

The N-alkyl-N-glycosyl product can be separated by removal of the solvent and precipitation from diethyl ether. The pure product can be separated using known precipitation methods.

A preferred embodiment of the third aspect of the present invention comprises a process for the preparation of an N-methyl-N-glycosyl derivative of a methyl ester of an antibiotic of a polyenemaprolylide group which is represented by formula (1) (b) The N-glycosyl derivative of the antibiotic of the polyenemarcholide group obtained by the rearrangement reaction is dissolved in a solvent, preferably a solution of the derivative of N, N-dimethylformamide in a solvent, preferably N, After conversion to a suspension by precipitation with ethyl ether, followed by treatment with an etheric solution of diazomethane at a reduced temperature, preferably at a temperature of from -5 ° C to + 5 ° C, stirring, And separating the crude product by precipitation from a concentrated solution with excess diethyl ether, and purifying the crude product according to a known method, wherein M in formula (1) (b) Refers to the residue of a group of antibiotics and R denotes the reaction of antibiotics with mono- or oligosaccharides, preferably D-glucose, L-glucose, D-mannose, D-galactose, lactose, or maltose, Means a portion of the sugar residue formed by the reaction.

In a fourth aspect of the present invention, MR, R ¹ and R ² are the first aspect of the present invention. Alkyl-N-glycosyl derivatives of alkyl esters of polyenemarcholide antibiotics in formula (2) as defined in the second and third aspects, wherein the method comprises the steps of homogeneously Suspending the N-alkyl-N-glycosyl derivative prepared according to the third aspect of the present invention in sufficient water to effect the formation of the suspension, acidifying the resulting suspension and isolating the product . The preparation of salts of N-methyl-N-glycosyl derivatives of methyl esters of polyene macrolide antibiotics is particularly preferred.

Organic or inorganic acids can be used for acidification of the suspension. L-aspartic acid is more preferable.

The separation of the pure product can be carried out by precipitation of the crude product with an organic solvent, followed by washing with further suitable solvents and drying. It is preferred that the solvent used to precipitate the crude product is miscible with water, preferably acetone. Typical solvents used to wash the product include acetone and diethyl ether. It is preferred that the product is dried under reduced pressure.

A preferred embodiment of the fourth aspect of the present invention comprises a process for the preparation of a salt of an N-methyl-N-glycosyl derivative of a methyl ester of an antibiotic of the polyene macrolide group represented by formula 2 (b) The N-glycosyl derivative of the antibiotic of the polyenmarmrolide group obtained by re-rearrangement may be isolated from a solution of the derivative in an organic solvent, preferably N, N-dimethylformamide, in a solvent, preferably diethyl ether To a suspension and then treated with an ethereal solvent of diazomethane at a reduced temperature, preferably -5 ° C to + 5 ° C, stirred, and the solvent evaporated and preferably dissolved in excess diethyl ether And the crude product is purified according to known methods, and then the derivative obtained as a solid is suspended in a small amount of water, and a stoichiometric Of an organic or inorganic acid and then the product is precipitated with an excess of an organic solvent miscible with water, preferably acetone, and then the solid is preferably washed with acetone, (B) wherein M represents a residue of an antibiotic substance of a polyenecrolide group, R represents a mono or oligosaccharide, and preferably, Refers to a portion of the sugar residues formed from the reaction of D-glucose, L-glucose, D-mannose, D-galactose, lactose, or maltose with an antibiotic, and concurrent potentiostatic reactions.

A fifth aspect of the present invention includes an N-alkyl-N-glycosyl derivative of formula (1) and salts thereof for use in therapy.

A sixth aspect of the present invention provides a method of treating a homogenous infection in humans and animals, including the administration of an N-alkyl-N-glycosyl derivative of formula (a) or a salt thereof as defined above do. N-methyl-N-glycosyl derivatives of formula (1) (b) and salts thereof are of particular interest.

A first preferred embodiment of the sixth aspect of the invention comprises a method of treating an exogenous and endogenous homologous infection in humans and animals, comprising the step of administering to the mammal an effective amount of a methyl esters of antibiotics of the polyenecrolide group represented by formula (I) N-methyl-N-glycosyl is widely used for the treatment of infections. In the above formula (1), M represents a residue of a polyene macrolide group, R represents a mono- or oligosaccharide, preferably D- Glucose, D-mannose, D-galactose, lactose, or a portion of the sugar residues formed by the reaction of the maltose with the antibiotic and the simultaneous potentiation reaction.

A second preferred embodiment of the sixth aspect of the present invention relates to a method of treating an exogenous and endogenous fungal infection in humans and animals comprising the step of administering to a mammal an effective amount of a methyl ester of an antibiotic of a polyenecrolide group represented by formula 2 (b) The salt of an N-methyl-N-glycosyl derivative is widely used for the treatment of infection. In the above formula (2), M represents a residue of an antibiotic substance of a polyenecrolide group, R represents a mono- or oligosaccharide, Means a part of sugar residues formed by the reaction of D-glucose, L-glucose, D-mannose, D-galactose, lactose, or maltose with antibiotics and concurrent potato potential reaction, A is an organic acid or an inorganic acid Means anion.

A seventh aspect of the present invention provides an N-alkyl-N-glycosyl derivative of formula (a) for use in the treatment of fungal infections and salts thereof as defined above. N-methyl-N-glycosyl derivatives of formula (I) or salts thereof are particularly interesting.

The infection provided with the treatment may be an internal infection or an external infection. The mode of administration will depend on the nature of the infection. Thus, the compounds of the present invention may be formulated for administration intravenously, intraperitoneally, orally, topically, subcutaneously, rectally or intravaginally.

An eighth aspect of the invention provides a composition for use in the treatment of a fungal infection, comprising an N-alkyl-N-glycosyl derivative of formula (a) or a salt thereof and a physiologically acceptable carrier . The nature of the N-alkyl-N-glycosyl derivative and the carrier will depend on the mode of administration. The compositions may optionally be formulated from one or more compounds according to the invention in combination with other antimicrobial agents known in the art, if desired. Compositions containing the N-methyl-N-glycosyl derivatives of formula (1) (b) and salts thereof are of particular interest.

A ninth aspect of the present invention is a pharmaceutical composition comprising, for use in pharmacy or veterinary medicine, at least one N-alkyl-N-glycosyl derivative of formula (a) or a salt thereof and a physiologically acceptable, To provide a unit dosage form. The unit dosage form may optionally be formulated from one or more compounds according to the invention in combination with other known antimicrobial agents. Of particular interest is the analogy with previous embodiments of unit dosage forms of the present invention containing an N-methyl-N-glycosyl derivative of Formula I (a).

The properties of the unit formulations mentioned above will depend on the mode of administration. Typically tablets and capsules are suitable for oral administration and creams and patches are suitable for topical administration with a paddle suitable for rectal and vaginal administration.

A tenth aspect of the present invention provides the use of an N-alkyl-N-glycosyl derivative of formula (I) and salts thereof for the manufacture of a medicament for use in the treatment of fungal infections. Compounds in which both N-alkyl and alkyl ester substituents are methyl groups are of particular interest.

Structural measurements performed on the compounds of the present invention using spectroscopy show that preservation of the parent antibiotic is maintained during the course of the reaction.

The invented method for the production of N-methyl-N-glycosyl derivatives of the methyl esters of antibiotics of the polyene macrolide groups provides the desired products without altering the parent antibiotic structure. The structure of the obtained compound was tested using spectroscopy. Proof is exemplified by measurement of the structure of N-methyl-N-D-fructosyl amphotericin B methyl ester, i.

The electronic spectrum of the N-methyl-ND-fructosyl amphotericin B methyl ester is the same as that of the parent antibiotic, meaning amphotericin B, and the method of the present invention does not induce the degradation of the polyene chromophore, It has been demonstrated that the extinction value (E ^1% _{1 cm} = 1300 at 382 nm) confirms the high purity of the obtained product. The infrared absorption spectrum of N-methyl-N-methyl-ND-fructosyl amphotericin B methyl ester indicates that the band related to the stretching vibration of the ester carbonyl group at 1730 cm ^-1 and the band free radical of the free carboxyl group, which means that the carboxyl group was completely converted to the methyl ester group . Complete information on the structure of N-methyl-ND-fructosyl amphotericin B methyl ester can be found in the nuclear magnetic resonance spectra (NMR) for ¹ H (DQF-COZY, ROESY), ¹³ C A heterocorrelated spectrum (mutation 3000 MHZ spectrometer) was provided that allowed partitioning of formula (3).

The most important ¹ H and ^{1 3} C information are listed in Tables 1 and 2, respectively. NMR data for aglycones of amphotericin B are reported in Magn. Reson. Chem. 30 , 275, (1992), P.Sowinski, J. Pawlak, E. Borowski, P. Garibol. ¹ H chemical shifts (in the DMSO / MeOD solvent system) of N-CH ₃ (δ = 2.35 ppm) and H-1 (2.03 and 3.15 ppm) are characterized by the effect of amino substituents. After acidification, delta changes to 2.92 ppm for N-CH ₃ and to 3.64 ppm for H-1 due to the protonation of the amino group (隆 for H-3 'changed to 3.19 ppm). This data is supported by the effect of the ROE between proton _{NCH 3 / H3 ', NCH 3} / H2', NCH 3 / H1b and 1a / H3 '. The coupling constant and ROE represent the ⁴ C ₁ form of the mycosamin moiety as previously found for the amphotericin B,

Table 1 shows chemical shifts and ROE effects of the disaccharide fragments of ¹ H and N-methyl-ND-fructosyl amphotericin B methyl esters.

Table 1.

proton δ [ppm] ROE for proton δ [ppm] ROE for proton Pyridine-d ₅ : methanol-d ₄ 9: 1 DMSOO-d ₆ : methanol-d ₄ 4: 6 1 '4.77 2', 3 ', 5', 18b, COOMe 4.48 3 ', 5'2' 4.41 1 ', 3', NMe, 17, COOMe, 4.04 3 ', NMe, COOMe 3' 2.06 1 ', 2', 5 ', NMe, 1 a, COOMe 1.89 1', 2 ', NMe 1 b 4' '3.45 1', 3 ', 6'6' 1.23 4 ', 5' 1.27 4 ', 5'1a 2.56 3', 3 (2 ') 2.301b 3.58 NMe 3.15 3, 3'3 4.41 1a? 3.65 1b4 4.76 3.975 4.41 3.636a 4.21 3.576b 4.36 3.76NMe 2.29 2 ', 3', 1b, COOMe 2.35 2 ', 3', 1a, 1b, COOMeCOOMe 3.70 1 ', 2', 3 ' ', NMe

Table 2 shows ¹³ C-NMR chemical shifts of the disaccharide fragments of N-methyl-ND-fructosyl amphotericin B methyl ester and their comparison with D-fructose data.

Table 2.

Tetra of Fructus ^* beta -D-fructopyranose alpha -D-furanos beta -D-fructorolanose carbon delta [ppm] delta [ppm] delta [ppm] delta [ppm] 1 '98.22' 72.43 '66.44' 69.85 '72.36' 18.21 62.2 64.1 62.1 63.92 98.1 99.1 105.1 102.43 66.9 70.5 83.0 76.54 71.2 68.4 77.0 75.55 72.2 70.0 82.2 81.56 64.6 64.7 62.1 63.3NMe 40.9COOMe 51.6

* See [SNRosenthal and JH Fendler, Progr. Phys. Org. Chem. 13 , 280 (1976).

Comparison of ¹³ C-NMR data from Table 2 and literature data for D-fructose on fructosyl fragments shows the pyranoside form of the sugar substituent. The observed coupling constants recorded in Table 3 demonstrate the boat shape of the fructopyranoside ring represented by Formula 4. [ Only this boat shape of the ring is exactly the same as the measured coupling constant.

Table 3 shows the coupling constants J _{H, H} for the modifier (pyridine-d ₅ : methanol-d ₄ 9: 1) of the disaccharide fragment of N-methyl-ND-fructosyl amphotericin B methyl ester, The coupling constants and chemical shifts of the coupled spin systems H3 to H6 were repeatedly refined by computer simulation.

Table 3.

proton J [Hz] quantum J [Hz] 1 ', 2'2', 3'3 ', 4'4', 5'5 ', 6' ~ 02.29.59.26.0 1a, 1b3, 44, 55, 6a5, 6b6a, 6b4, 6a 11.61.058.095.763.61-10.86-0.1

The results confirm the morphology of the fructosyl fragment of the N-methyl-N-D-fructosyl amphotericin B methyl ester represented by formula (4). The different N-methyl-N-glycosyl derivatives of the methyl esters of antibiotics of the polyene macrolide groups are characterized by methods similar to those described above.

The present invention will now be described with reference to the following non-limiting embodiments. Further embodiments within the scope of the invention will be apparent to those skilled in the art.

Anti-fungal activity

Toxicity and antimicrobial activity in in vitro experiments were measured for all of the manufactured compounds. Activity and toxicity in vivo were also measured for compounds with the best properties, namely N-methyl-N-D-fructosyl amphotericin B methyl ester asparate.

Antimicrobial activity in vitro

The antimicrobial activity of the compounds was determined according to the standard method for the polyene macrolide method. 10 ⁴ cells / ml of the Candida albicans organism Candida albicans ATCC 262778 to be tested in the Sabouraud liquid medium was incubated with the tested antibiotics (serially diluted) for 24 hours at 30 ° C. Amphotericin B was used as a reference material. The compound was dissolved in DMF and an appropriate amount of solution was added to the medium. The turbidity titration method (660 nm) was used to measure the extent of growth inhibition. The concentration of antibiotics inhibited by 50% growth of fungi was measured by a dose response curve. The IC ₅₀ values obtained are characteristic of the antimicrobial activity of the compounds.

Toxicity in vitro

The toxicity of compounds in animal experiments in animal cells was determined by measuring the extent of hemolysis in human erythrocytes using the standard method for the polyenemarcholide method. Human red blood cells isolated from fresh and citrated human blood were washed twice with cold saline. Cells were diluted 250 fold with saline and equilibrated at 37 ° C for 30 min. Samples of erythrocytes were incubated with various concentrations of antibiotics (base solution in DMP) for 30 minutes at 37 占 폚. After centrifugation, dissolution of erythrocytes was evaluated by measuring the hemoglobin released from the solution. The optical density of the supernatant was measured at 550 nm. The results are shown as the EH ₅₀ value as the concentration of the antibiotic substance in which hemolysis occurred at 50%. The value of EH ₅₀ was interpreted as a curve showing the degree of haemolysis according to antibiotic dose.

Toxicity in vivo experiments

The in vivo toxicity of the methyl ester of N-methyl-ND-fructosyl amphotericin B L -aspartate was measured as the maximum tolerated dose (MTD) and acute toxicity (LD ₅₀ ). To determine the MTD dose, the compounds were dissolved in 5% glucose solution and single and multiple doses were administered intravenously and intraperitoneally to Balb / c mice. The maximum single dose was 100 mg / kg administered intravenously and more than 200 mg / kg administered intraperitoneally. The maximum combined dose given at 100 mg / kg per day for 5 days was even higher. For this dose, toxic effects were not observed for 20 days.

The LD ₅₀ of the acute poison, the methyl ester of N-methyl-ND-fructosyl amphotericin B L-aspartate, was measured with a Swiss Webster male mouse with an average weight of 20 g. Various doses of amphotericin B were administered intravenously in animals in the form of tested compounds dissolved in 5% glucose and in the form of fonns for comparison. The administered volume of solution is 0.5 ml. 0.5 ml of a 5% glucose solution was administered to the mice as a reference. All doses of these two preparations were administered to 5 mice. Animals were observed for 7 days. Next, after the animal died, some serum indicators were measured. Increased levels of aspartate aminotransferase or creatinine were not observed when compared to the control. In the methyl ester of N-methyl-ND-fructosyl amphotericin B L -aspartate, the LD ₅₀ was 400 mg / kg, whereas the amphotericin B in the form of the fonnzide was 6 mg / kg.

Chemotherapy efficacy

The chemotherapeutic efficacy of the methyl ester of N-methyl-ND-fructosyl amphotericin B L-aspartate was measured using the myelin candy dose model of the whole body. Candida albicans were cultured in Sabouraud dextrose fermentation broth overnight at room temperature. Fungal cells were centrifuged, washed twice with 0.9% sodium chloride solution, and suspended in physiological salt solution. 10 ⁵ Candida cells in 0.2 ml of 0.9% sodium chloride solution were injected intravenously into a female Swiss Webster mouse weighing 25 g. Initially, the infection was systemic, but 2 to 3 days were concentrated in the kidneys. Unspecified animals generally died 7 to 14 days post-infection. On post-infection day 3, animals were treated intravenously twice daily for five consecutive days, between five and six hours. The preparation was administered as a 5% glucose solution. The animals were observed for 5 weeks from the day of infection. After this time, the surviving animals were sacrificed, their kidneys were removed, homogenized in sterile water, the bacteria were applied to Sabouraud dextrose agar, and the number of grown colonies of Candida was counted. The efficacy of the chemotherapy regimen in which the test was shown to be 50% survival of the animal and the removal of Candida from the kidneys in half of the mice was expressed as a single dose of mg / kg. One time dose, ED _50, was determined as described in J.HYG. 27 , 493, (1938). The ED ₅₀ value for the methyl ester of N-methyl-N-methyl-ND-fructosyl amphotericin B L -aspartate is 2.3 mg / kg based on survival and 6 mg / kg based on elimination in the kidney.

Manufacturing Example

N-methyl-N-D-glycosyl derivatives of methyl esters of antibiotics of polyene glycolide groups, their salts and their preparation methods are illustrated in the following examples.

Example I

Amphotericin B (E ^1% _{1 cm} = 1350 at 382 nm, MeOH) was dissolved in 15 ml of N, N-dimethylformamide, 0.3 g of D-glucose was added and the mixture was incubated at 37 ° C for 40 hours And stirred in a dark room. The reaction mixture was then cooled and the solid was precipitated with excess diethyl ether. The solid was centrifuged, washed twice with diethyl ether, and dried under reduced pressure. To remove excess glucose, the solid was suspended in 20 mL water, centrifuged, washed twice with a small amount of water, twice with acetone, and then twice with diethyl ether. The product was dried under reduced pressure to yield 0.98 g of ND-Flutosyl amphotericin B (E ^1% _{1 cm} = 1200, MeOH at 382 nm). The product was dissolved in 10 ml of N, N-dimethylformamide with stirring, and 50 ml of diethyl ether was added to the solution to obtain a good suspension. The suspension was cooled in ice to 0 ° C to 2 ° C and a diethyl ether solution of 2.5 moles diazomethane per 1 mole of the freshly prepared ND-fructo-amphotericin B was added under vigorous stirring. The reaction was carried out by thin film chromatography on silica gel in a solvent system with a volume ratio of chloroform-methanol-water of 10: 6: 1 v / v. After a complete run of the reaction carried out for about 2 hours, excess diazomethane and diethyl ether were evaporated under reduced pressure at temperatures not higher than 40 ° C. The crude product was precipitated from the residue with excess diethyl ether, centrifuged, washed twice with diethyl ether, then with n-hexane and dried under reduced pressure to give 0.95 g ≪ / RTI > Pure N-methyl-N-methyl-N-methyl-N-methyl-N-methyl-N-methyl-N-methyltoluenesulfonate amphotericin B methyl ester was prepared from the crude product in a solvent system with a volume ratio of chloroform-methane- water of 20: 8: 1 v / v to Merck Silicagel 60, And separated by column chromatography on a mesh. Thus, 0.95 g of the crude product is suspended in the mixture of the specific solvent, and if the product is not well dissolved, the volume ratio of the same solvent is changed to 10: 6: 1. The inferred fraction is centrifuged and the supernatant is loaded on a chromatography column and then developed into the recorded solvent mixture (except 20: 8: 1 v / v ratio). The eluate was analyzed on a silica plate using a solvent system with a volume ratio of chloroform-methanol-water of 10: 6: 1 v / v. The plate was visualized with a cerium sulfate reagent. A fraction of R _f = 0.5 to 0.54 containing the pure N-methyl-N-fructosyl derivative of amphotericin B methyl ester was collected. The combined fractions were evaporated under reduced pressure. The dry residue was dissolved in a small amount of N, N-dimethylformamide, the product was precipitated with excess diethyl ether, the solid was centrifuged, washed twice with diethyl ether and dried in a vacuum drier. 0.137 g of N-methyl-ND-fructosyl amphotericin B methyl ester of E ^1% _{1 cm} = 1300 on 382 nm in methanol was obtained. Proof of the structure is recorded in the above-mentioned section. The antimicrobial activity of the compound against Candida albicans measured by the method described above provides IC ₅₀ = 0.12 / / ml, and the toxicity to human erythrocytes measured by the method described above is higher than 350 / / ml EH ₅₀ values were obtained. In comparison, the EH ₅₀ value for the starting amphotericin B was 1.5 μg / ml. The exact value of EH ₅₀ for N-methyl-ND-fructosyl amphotericin B methyl ester as 350 μg / ml of said insoluble compound under the conditions of the experiment could not be determined.

Example II

0.5 g of Candida (E ^1% _{1 cm} = 1175 in MeOH) at 382 nm and 0.15 g of D-glucose were dissolved in 10 ml of N, N-dimethylformamide and stirred at 37 캜 for 36 hours. Also carried out analogously to the method of Example I, 0.43 g of ND-fructosylcandidine was used to elute E ^1% _{1 cm} = 1100 in MeOH at 382 nm. The product was methylated with diazomethane in diethyl ether analogously to Example I to provide 0.4 g of crude product. Pure N-methyl-ND-fructosyl candidin methyl ester was isolated by column chromatography, i. E. Analogous to the method provided in Example I. Fractions containing pure derivatives are characterized by R _f = 0.49-0.52 on thin layer chromatography. The fractions were collected, evaporated to dryness, dissolved in a small amount of N, N-dimethylformamide and the solid was precipitated with diethyl ether. Separating the solid centrifuge, washed with diethyl ether and dried in a vacuum dryer N- methyl -ND- fructosyl candy Dean methyl ester 0.05g, MeOH 382nm in _{^{E 1% 1㎝ = 1200, IC}} 50 = 0.75㎍ of / Ml and 300 μg / ml or more of EH ₅₀ were obtained. The structure of the compound was determined by the same method for the structure of N-methyl-ND-fructosyl amphotericin B methyl ester.

Example III

2 g of Nistatin (E ^1% _{1 cm} = 870 at 304 nm in MeOH) and 0.6 g of D-glucose were dissolved in 35 ml of N, N-dimethylacetamide and stirred at 37 ° C for 40 hours. After complete reaction, the crude product was precipitated with diethyl ether, centrifuged and dried under vacuum. The excess glucose was then washed with a small 1: 1 mixture of water-acetone ratio, then with acetone, diethyl ether, centrifugation of the product and drying in a vacuum dryer to yield ND-Fructosylcystin 1.2 g of E ^1% _{1 cm} = 720 at 340 nm in NeOH). The product was methylated using diazomethane in Example I to give 1.25 g of crude product. The product was purified on a silica gel column in Example I. Fractions containing pure N-methyl-ND-fructosylnistatin methyl ester are characterized by R _f = 0.49-0.52 on thin layer chromatography in Example I. The fractions were collected and evaporated to dryness at 30 < 0 > C under reduced pressure, dissolved in a small amount of N, N-dimethylformamide and the product precipitated with diethyl ether. The product was centrifuged, washed with diethyl ether and dried in a vacuum oven to give 0.17 g of N-methyl-ND-fructosylnastatin methyl ester; E ^1% _{1 cm} = 900 at 304 nm in MeOH. The structure of the compounds was determined by the method described for the N-methyl-ND-fructosyl amphotericin B methyl ester. IC ₅₀ = the 6.8㎍ / ㎖ large and EH ₅₀ of the compound obtained than 300㎍ / ㎖ was found.

Example IV

0.79 g of the major constituent (E ^1% _{1 cm} = 900 at 378 nm in MeOH) of the basidiin, namely the antibiotic complex Oreofacin in 15 ml of N, N-dimethylacetamide and 0.22 g of D- Lt; / RTI > After complete reaction, the mixture was allowed to cool and the solid was precipitated with excess diethyl ether. The solid was centrifuged, washed with diethyl ether and dried under reduced pressure to give 0.8 g of initial ND-fructosylcysteine (E ^1% _{1 cm} = 720 at 378 nm in MeOH). The resulting derivative was methylated with diazomethane as in Example I to give 0.5 g of crude product. Pure N-methyl-ND-fructosylbasdine methyl ester was separated by chromatography on silica gel, analogous to that described in Example I, but the column was developed using a 30: 8: 1 solvent system with a chloroform-methanol- I did. The fractions containing pure derivatives of bacidin and having a thin film chromatographic value of R _f = 0.53-0.5 in a chloroform-methanol-water ratio 13: 8: 1 solvent system were combined and combined. The method was also carried out by the method in Example III. 0.07 g of N-methyl-ND-fructosylbacydin methyl ester (E ^1% _{1 cm} = 900 at 378 nm in MeOH). The compounds exhibited IC ₅₀ = 0.01 μg / ml and EH ₅₀ = 170 μg / ml. The structure of the compound was determined by the method described for N-methyl-ND-fructosyl amphotericin B methyl ester.

Example V

0.79 g of the major constituents of candicidin D, the antibiotic complex candicidin, were treated in the same manner as described in Example IV. N-methyl-ND-fructosyl Candididin D methyl ester was obtained; E ^1% _{1 cm} = 920 at 378 nm in MeOH under the conditions described in Example IV, and R _f of the compound in thin film chromatography was 0.50-0.53. The compounds showed IC ₅₀ = 0.01 μg / ml and EH ₅₀ = 180 μg / ml. The structure of the compounds was determined by the method described in N-methyl-ND-fructosyl amphotericin B methyl ester.

Example VI

0.79 g of the major constituent of the antibiotic compound tricomycin was treated in the same manner as described in Example IV. Yielded 0.1 g of N-methyl-ND-fructosyl tricomycin methyl ester, E ^1% _{1 cm} = 910 at 378 nm in MeOH under the conditions described in Example IV, R _f of the compound in thin- 0.49 to 0.52. The compounds exhibited IC ₅₀ = 0.013 μg / ml and EH ₅₀ = 165 μg / ml. The structure of the compounds was determined by the method described in N-methyl-ND-fructosyl amphotericin B methyl ester.

Example VII

0.5 g of amphotericin B (E ^1% _{1 cm} = 1350 at 382 nm in MeOH) and 0.15 g of L-glucose in 8 ml of N, N-dimethylformamide were stirred at 37 캜 for 40 hours. Thereafter, it was operated as in Example I. 0.065 g of N-methyl-NL-fructosyl amphotericin B methyl ester was obtained; E ^1% at 382 nm in MeOH = ¹ _cm = 1280. Compounds showed values greater than IC ₅₀ = 0.42 μg / ml and EH ₅₀ = 200 μg / ml. The structure of the compound was determined by the method described in the methyl ester of N-methyl-ND-fructosyl amphotericin B.

Example VIII

0.52 g amphotericin B (E ^1% _{1 cm} = 1350 at 382 nm in MeOH) and 0.153 g D-mannose were dissolved in 10 ml N, N-dimethylformamide and stirred at 37 ° C for 40 hours. Thereafter, it was operated as in Example I. The pure derivative was isolated by silica gel column chromatography in a similar manner to Example I and the fraction with a value of R _f = 0.53-0.55 in thin film chromatography run according to the method described in Example I was pooled. Thereafter, it was operated as in Example I. 0.08 g of N-methyl-ND-fructosyl amphotericin B methyl ester was obtained: E ^1% _{1 cm} = 1280 at 382 nm in MeOH. The compound exhibited an IC ₅₀ = 0.42 g / ml and an EH ₅₀ of 165 g / ml or more. The structure of the compounds was determined by the method described in N-methyl-ND-fructosyl amphotericin B methyl ester.

Embodiment IX

0.5 g Amphotericin B (E ^1% _{1 cm} = 1350 at 382 nm in MeOH) and 0.3 g D-lactose in 12 ml N, N-dimethylformamide were stirred at 37 캜 for two days. Thereafter, it was operated as in Example I. The pure derivative was isolated by silica gel column chromatography in a similar manner to Example I and fractionated with thin layer chromatography with R _f = 0.25-0.30, operating according to the method described in Example I. 0.08 g of N-methyl-Nd-fructosyl-galactosyl amphotericin B methyl ester was obtained: E ^1% _{1 cm} = 1000 at 382 nm in MeOH. The compounds exhibited IC ₅₀ = 6.0 μg / ml and EH ₅₀ greater than 200 μg / ml.

Example X

0.5 g of the N-methyl-N-methyl-N-methyl-N-methylated porphyrin B methyl ester prepared in accordance with Example I was suspended in 10 ml of water and 2 ml of water containing 0.059 g of aspartic acid dissolved therein was added. To make the solution effective, the acidic solution was added drop wise while stirring. The solution was filtered to remove a small amount of residual solids and excess acetone was added to the clear filter until all salts were precipitated. The solid was filtered or centrifuged, washed twice with acetone, washed twice with diethyl ether, dried under reduced pressure and eluted with ^1% _{1 cm} = 1100 N-methyl-ND-fructose at 382 nm in MeOH 0.5 g of methyl ester of tosyl amphotericin B L -aspartate was obtained. Under the conditions of Example I, thin film chromatography gave _Rf = 0.5 to 0.54. The product is soluble in a 5% aqueous solution of N, N-dimethylformamide, dimethylsulfoxide, and glucose. It is more soluble in water. The compound exhibited an IC ₅₀ = 0.125 μg / ml and an EH ₅₀ greater than 350 μg / ml.

Claims (43)

Alkyl-N-glycosyl derivatives of alkyl esters of antimicrobials of the macrolide group of formula (1): In this formula, M means the residue of an antibiotic of the polyene macrolide group, R means a part of the sugar residue formed by the reaction of an antibiotic with a mono or oligosaccharide, R ¹ and R ² are the same or different and each represents a C _1-4 alkyl group. The composition according to claim 1, wherein M is selected from amphotericin B, candidin, candidone, candidin, maikoheptin, nystatin, polypugjin, oreofacin, barcidin, tricomycin or candicidin N-alkyl-N-glycosyl derivatives of alkyl esters of antibiotics of polyene macrolides. 3. The method according to claim 1 or 2, wherein the mono or oligosaccharide which derives the sugar residue R is selected from D-glucose, L-glucose, D-mannose, D-galactose, lactose or maltose. N-alkyl-N-glycosyl derivatives of alkyl esters of antibiotics in lead groups. Claim 1 to claim according to any one of 3, wherein, R ¹ and R ² is N- alkyl -N- glycosyl derivatives of alkyl esters of antibiotics of the polyene macrolide group, characterized in that the methyl group. A salt of an N-alkyl-N-glycosyl derivative of an alkyl ester of an antibiotic of a polyenmarmrolide group of formula (2) In this formula, M, R, R ¹ and R ² are as defined in any one of claims 1 to 4, A means an anion of an organic or inorganic acid. 6. A salt of an N-alkyl-N-glycosyl derivative of an alkyl ester of an antibiotic of a polyenemarcholide group according to claim 5, wherein A is an anion of L-aspartic acid. 5. A method according to any one of claims 1 to 4, characterized in that a simultaneous pyrroelectric potential reaction takes place to react the polyenmarmrolide antibiotic with a mono- or oligosaccharide to produce a polyenemalcolide antibiotic N-glyco Obtaining a silical derivative; Possibly separating the product of the rearrangement reaction; A method for preparing an N-alkyl-N-glycosyl derivative of an alkyl ester of an antibiotic of a polyenmarmrolide group, comprising the steps of treating the product with an alkylating agent and purifying the crude product. 8. A process according to claim 7, characterized in that the product of the potentiostatic reaction is separated in the form of a suspension by precipitation from a solution in which the potential reaction takes place. 9. Process according to claim 7 or 8, characterized in that N, N-dimethylformamide is used to support the potenial reaction. 10. A process according to any one of claims 7 to 9, characterized in that the diethyl ether is used to effect the formation of a suspension by precipitation of the product of the potentiostatic reaction. 11. Process according to any one of claims 7 to 10, characterized in that the alkylation of the product of the potentially rearrangement reaction is carried out at a reduced temperature. 11. Process according to claim 10, characterized in that the alkylation is carried out at a temperature of from -5 [deg.] C to +5 [deg.] C. 13. A process according to any one of claims 7 to 12, characterized in that the alkylating agent is an etheric solution of diazomethane. 14. A process according to any one of claims 7 to 13, which comprises suspending an N-alkyl-N-glycosyl derivative prepared in sufficient water to effect the formation of a homogenous suspension, acidifying the resulting suspension, Alkyl-N-glycosyl derivative of an alkyl ester of a polyenmarmrolid antibiotic of formula (2) according to claim 5 or 6, 15. The method according to claim 14, wherein the organic acid or inorganic acid is used for acidifying the suspension. 16. The method according to claim 15, wherein the L-aspartic acid is used in the acidification of the suspension. 17. A process according to any one of claims 14 to 16, characterized in that the separation of the pure product is carried out by precipitating the crude product with an organic solvent, followed by washing with further suitable solvents and drying. ≪ / RTI > salt of an alkyl ester of a substance. 18. A salt of an alkyl ester of polyenmarmrolide antibiotics according to any one of claims 14 to 17, characterized in that the solvent used to precipitate the crude product is converted to water. N-Alkyl-N-glyco A method for producing a silical derivative. 19. The method according to claim 18, wherein the solvent is acetone. A method for preparing a salt of an alkyl ester of a polyenmarmrolide antibiotic, characterized in that the solvent is acetone. 20. A salt of an alkyl ester of a polyenmarmrolide antibiotic according to any one of claims 17 to 19, characterized in that the further suitable solvent used for the washing is selected from acetone and diethyl ether. -Alkyl-N-glycosyl derivatives. 21. A process as claimed in any one of claims 14 to 20, wherein the product is dried under reduced pressure. A process for the preparation of a salt of an alkyl ester of a polyenmarmrolide antibiotic, characterized in that the product is dried under reduced pressure. 7. An N-alkyl-N-glycosyl derivative of formula (I) according to any one of claims 1 to 6 for use in therapy or a salt thereof. 7. A method according to any one of claims 1 to 6 for the treatment of a homogeneous infection in humans and animals characterized by the administration of an N-alkyl-N-glycosyl derivative of formula (1) How to. 7. A method according to any one of claims 1 to 6 for the treatment of a homogeneous infection in humans and animals characterized by the administration of an N-methyl-N-glycosyl derivative of formula (a) Way. 7. A pharmaceutical composition according to any one of claims 1 to 6, characterized in that it comprises a N-alkyl-N-glycosyl derivative of formula (1) or a salt thereof and a physiologically acceptable carrier. A composition for use in treating an infection. Characterized in that it comprises at least one N-alkyl-N-glycosyl derivative of formula (1) or a salt thereof and a physiologically acceptable carrier, formulated for use in pharmacy or veterinary medicine Formulation. Characterized in that the N-alkyl-N-glycosyl derivative of the formula (1) or a salt thereof comprises an N-methyl-N-glycosyl derivative of the methyl ester of the formula (1) 26. A composition for use in treating a homogeneous infection according to claim 25 or a unit dosage form according to claim 26. 28. A composition or unit dosage form according to any one of claims 25 to 27, further comprising a known antimicrobial agent. 29. A composition or unit dosage form according to any one of claims 25 to 28, characterized in that it is formulated for administration intravenously, intraperitoneally, orally, topically, subcutaneously, rectally or intravaginally. 7. A method according to any one of claims 1 to 6 for the preparation of a medicament for use in the treatment of a homogeneous infection, comprising administering an N-alkyl-N-glycosyl derivative of the alkyl ester of formula (I) How to use. 31. The method of claim 30, wherein all N-alkyl and alkyl ester substituents are methyl groups. 7. The method according to any one of claims 1 to 6, wherein an N-alkyl-N-glycosyl derivative of formula (1) or a salt thereof is used for the treatment of a homogenous infection. 33. The method of claim 32, wherein the N-alkyl and alkyl ester substituents are all methyl groups. 34. The method according to claim 32 or 33, wherein the homogenous infection is exogenous or endogenous. N-methyl-N-glycosyl derivatives of methyl esters of antibiotics of the polyenemarcholide group of formula (1): In this formula, M means the residue of an antibiotic of the polyene macrolide group, R refers to a portion of the sugar moiety formed by the reaction of a mono or oligosaccharide, preferably D-glucose, L-glucose, D-mannose, D-galactose, lactose or maltose with an antibiotic and concurrent potato potential reaction. 38. The method of claim 37, wherein the antibiotic of the polyenmarmrolide group is selected from the group consisting of amphotericin B, candidin, candidoxin, candidinin, maikoheptin, nystatin, polypuggin, oreofacin, Lt; RTI ID = 0.0 > candycidine. ≪ / RTI > A salt of an N-methyl-N-glycosyl derivative of an antibiotic of a polyenemarcholide group represented by the following formula (2) In this formula, M means the residue of an antibiotic of the polyene macrolide group, R is a part of the sugar moiety formed by the reaction of a mono or oligosaccharide, preferably D-glucose, L-glucose, D-mannose, D-galactose, lactose or maltose with an antibiotic and concurrent potato potential reaction. 40. The method of claim 39, wherein the antibiotic of the polyenemedrolide group is selected from the group consisting of amphotericin B, candidiene, candidone, candidinin, maikoheptin, nystatin, polypuggin, aureopacin, ≪ / RTI > 41. A salt according to claim 39 or 40, wherein A is associated with an anion of L-aspartic acid. An N-methyl-N-glycosyl derivative of a methyl ester of an antibiotic of the polyenemarcholide group represented by the formula (1) (wherein M represents a residue of an antibiotic substance of a polyenemarcholide group, R means a part of the sugar residue formed by the reaction of a mono or oligosaccharide, preferably D-glucose, L-glucose, D-mannose, D-galactose, lactose or maltose and concurrent potato potential reaction) As a method, the N-glycosyl derivative of the antibiotic of the polyenmarmrolide group obtained by preselection reaction is dissolved in an organic solvent, preferably a solution of a derivative in N, N = dimethylformamide in a solvent, Diethyl ether to give a suspension which is then treated with an etheric solution of the diazolmethane at a reduced temperature, preferably at -5 to + 5 < 0 > C, stirred, Advantageously, and separated by precipitation from the concentrated solution by an excess of diethyl ether, the method comprising the step of purifying the crude product according to known methods. An N-methyl-N-glycosyl derivative of a methyl ester of an antibiotic of the polyenemarcholide group represented by the formula (1) (wherein M represents a residue of an antibiotic substance of a polyenemarcholide group, R means a part of the sugar moiety formed by the reaction of a mono or oligosaccharide, preferably D-glucose, L-glucose, D-mannose, D-galactose, lactose or maltose with an antibiotic, There is provided a method for producing an N-glycosyl derivative of an antibiotic of a polyenmarmrolide group obtained by a potenial reaction, preferably in an organic solvent, preferably a solution of a derivative in N, N-dimethylformamide, Is converted to a suspension by precipitation with diethyl ether followed by treatment with an etheric solution of diazomethane at reduced temperature, preferably at -5 to + 5 < 0 > C, Preferably by precipitation from a concentrated solution with an excess of diethyl ether, purifying the crude product according to known methods, then suspending the derivative obtained as a solid in a small amount of water and adding a stoichiometric amount And then the product is precipitated from the resulting solution with water, preferably an excess of an organic solvent miscible with acetone, the solids are preferably washed with acetone, preferably with di Washing with ethyl ether and drying preferably under reduced pressure. N-methyl-N-glycosyl derivative of a methyl ester of an antibiotic of the polyene macrolide group represented by the formula (1) (wherein M represents a residue of an antibiotic substance of a polyene macrolide group, R means a part of the sugar moiety formed by the reaction of a mono or oligosaccharide, preferably D-glucose, L-glucose, D-mannose, D-galactose, lactose or maltose with an antibiotic, A method for treating an exogenous < RTI ID = 0.0 > 7 < / RTI > and endogenous homologous infection in humans and animals, including those used in the treatment of infection. An N-methyl-N-glycosyl derivative of a methyl ester of an antibiotic of a polyenemarcholide group represented by the formula (2) (wherein M represents a residue of an antibiotic substance of a polyenemarcholide group, R means a part of sugar residues formed by the reaction of mono or oligosaccharide, preferably D-glucose, L-glucose, D-mannose, D-galactose, lactose or maltose with antibiotics, A means an organic or inorganic acid) is used in the treatment of an infection. A method for treating an exogenous and endogenous homogeneous infection in humans and animals.