CS235343B2 - Method of 2%-aminotylosine derivatives production - Google Patents

Abstract

The subject of the invention is a production method of new macrolide antibiotics from the group 20-amino tylosin derivative and the like macrolide substances that are effective against bacteria and mycoplasmas.

Description

The present invention relates to a process for the production of a macrolide antibiotic, in particular to novel compounds which are similar to the known tylosin drug, as described, for example, in Tetrahedron Letters, 2339 (1970) and US Patent 3,178,341. .

Although tylosin is a valuable substance, there is still a need to develop additional antibiotics, both because of the need to obtain derivatives even more potent or with an even broader spectrum of activity, and because microorganisms easily develop resistance. However, chemical modifications of macrolide antibiotics of the tylosin type are very difficult. For this reason, most researchers are trying to find new microorganisms that produce related compounds.

It has now been unexpectedly found that the aldehyde group at the 20-position in tylosin-like systems can be converted by a reductive scanning amine to an amino group without simultaneously disrupting the macrolide system. Moreover, these derivatives have very good antibiotic activity.

The present invention therefore provides a process for the preparation of a novel macrolide antibiotic of formula (I)

where

R is hydrogen, hydroxyl or mycinosyloxyacin, Q is hydrogen,

X represents a group of the general formula

wherein (i) R ¹⁰ and R ¹¹ are independently hydrogen, C 1 -C 4 alkyl or C 3 -C 10 cycloalkyl or (CH 2) -Ph wherein q is an integer of 0, 1 or 2a /

Ph represents a phenyl radical optionally substituted by a (C 1 -C 4) alkoxycarbonyl, amino, halogen atom, (C 1 -C 4) alkyl or alkoxy radical, (ii) a single R ¹⁰ or R ¹ 'is a radical of formula

wherein r '2 represents a hydrogen atom or an alkyl radical of 1 to 4 carbon atoms, the other being as defined in (i) or (iii)

R ¹⁰ a

R ¹¹ forms together with the nitrogen atom to which they are attached a heterocyclic ring of 5 to 7 atoms, the other heteroatom being oxygen or nitrogen, as well as pharmaceutically acceptable salts and acyl esters, which are potent antibiotics.

Although no athereochemical configuration is given for the above structural formula, it is apparent that the stereochemistry is the same as that of tylosin. Aminosugar is wylaminosis.

The term "C 3 -C 10 cycloalkyl" refers to C 3 -C 10 carbon ring groups such as cyclobutyl, cyclohexyl, cyclooctyl and adamantyl. Similarly, C 1 -C 4 alkyl means straight or branched C 1 -C 4 alkyl radicals such as methyl, ethyl, n-propyl and isopropyl.

Preferred are compounds of formula I wherein R is myciaosyloxy.

When κ'θ and r '' form a heterocyclic ring of 5 to 7 atoms with oxygen or nitrogen as another heteroatom, the piperazine or morpholine moieties are typical examples. The heterocyclic ring must contain two heteroatoms, preferably saturated.

Another group of interesting compounds which may be mentioned are those in which X is amino, dimethylamino, aniline, H-methylaniline, o-ethoxycarbonylaniline, benzylamino, aminobenzylamino, halobenzylamino, cyclohexylamino and morpholine.

Preferred compounds obtainable by the process of the invention include:

20-deoxo-20- (N-benzylamino) demycarosyltylosin a

20-Deoxo-20- (N-morpholino) demycarosyltylosin and their pharmaceutically-acceptable ^- salts and acyl esters.

The hacrolides of formula I contain amino groups at the 20 and 3 'positions and can form acid addition salts. These salts, when non-toxic, can be used in chemotherapy in warm-blooded animals, for example, pharmaceutically acceptable salts can be used as antibiotics.

Typical salts of this type are those formed by conventional reactions with organic and inorganic acids such as sulfuric, hydrochloric, phosphoric, acetic, oxalic, citric, lactic, maleic, fumaric, palmitic, bile, pamoic, mucic, D-glutamic, d. - camphor, glutaric, glycolic, violet, tartaric, formic, lauric, stearic, salicylic, methanesulfonic, benzenesulfonic, sorbinic, picrous, benzoic and cinnamon.

Pharmaceutically acceptable acyl esters are preferably those derived from a monocarboxylic acid having 2 to 7 carbon atoms, for example acetic, proponic, butyric or isovaleric acid.

Compounds of formula I wherein Q 'is mycarosyloxy can be obtained by reductive amination of an aldehyde of formula II

where

R is as defined above, and

Q 'is mycarosyloxy using an amine reagent of formula III

HN (III)

JI kde

R < ¹ > and R < ¹¹ > are as defined above, or some of its acid addition salts,

The reductive amination is preferably carried out using a cyanoborohydride of the formula MBH3CN in which U is a metal of group Ia or an ammonium ion, with sodium cyanoborohydride being the most preferred.

An inert polar organic solution is usually employed as the solvent and for the reaction. a solvent, for example an alkanol of 1 to 4 carbon atoms, preferably methanol. Typical reaction temperatures range from 0 to 60 ° C, preferably 20 to 40 ° C. The reaction is usually carried out at a neutral pH in the range of 6-8.

The reaction is usually carried out using an excess of the reactant of formula (III), usually running at 2 to 3 equivalents. Dehydrating agents, such as molecular sieve 4A or anhydrous sodium or magnesium sulfate, are also preferably used in the reaction.

Reductive amination can also be carried out by reducing the aldehyde function at position 20 to the corresponding hydroxymethyl derivative. The hydroxymethyl derivative obtained is then modified by converting the hydroxymethyl group to a -CHgL group in which L is a readily cleavable group which can be readily removed by treatment with an amine reagent of formula III. For example, according to one embodiment of the process of the present invention, the hydroxyl group at the 20-position can be converted to a trifluoromethanesulfonyl group which can optionally be further converted and replaced with an iodine atom. According to a second embodiment, the iodo derivative may be formed directly by the addition of iodine dissolved in a suitable solvent such as dimethylformamide to a solution of the 20-hydroxymethyl derivative and triphenylphosphine under a nitrogen atmosphere.

The readily cleavable group at position 20, iodine, trifluoromethanesulfonyl, and the like, can then be replaced by reaction with an amine reagent in a suitable organic solvent, for example acetonitrile, to give a compound of formula I.

. Hydrolysis of compounds of formula I wherein Q is mycarose and / or R is mycinoeyloxy is carried out in a conventional manner as described in U.S. Patent 3,459,853.

For example, the mycarose may be hydrolytically cleaved at a pH of less than 4, preferably at a pH of 0.5 to 2.0 at a temperature of 0 to 60 ° C, preferably at room temperature.

The hydrolytic cleavage can be effected by treatment with a strong aqueous solution of an inorganic acid, for example hydrochloric or sulfuric acid, or a strong organic acid, for example p-toluenesulfonic acid.

Similarly, the remainder of mycinosis can also be hydrolytically cleaved, although less mild conditions must be used and care should be taken not to disrupt the macrolide system. A longer reaction time should be used and the pH maintained between 1.5 and 2.5. It is also desirable to use higher temperatures in the range of 80 to 130 ° C, preferably 90 ° C to the reflux temperature of the reaction mixture. Because mycinosis is more difficult to cleave, a compound of formula I in which R is a hydroxyl group is most preferably prepared by reductive amination of the starting compound of formula II in which R is a hydroxy group.

If desired, the compounds of formula I may be esterified to the acyl ester derivatives by treatment with acylating agents by conventional means. Suitable organic solvents for this use are pyridine and triethylamine.

The acylating agent may be an activated carboxylic acid derivative, for example a carboxylic acid anhydride or halide of 2 to 7 carbon atoms. In a similar manner, pharmaceutically acceptable salts of the above antibiotics can be obtained by converting them to these salts in a known manner.

Accordingly, the present invention provides a process for the preparation of a macrolide antibiotic of formula (I), a pharmaceutically acceptable salt or acyl ester thereof, which comprises:

(a) cleaves the carbohydrate sugar from a markolic antibiotic of formula I 'wherein Q' is mycarose by acid hydrolysis to form a compound of formula 1 wherein Q is hydrogen; and

b) optionally esterifying and / or converting the resulting product into a salt.

The aldehydes of the formula are known compounds and their preparation is disclosed, for example, in U.S. Patent Nos. 3,117,341, 4,332,361 and 4,321,362.

The compounds produced by the process of the invention are effective against Gram positive microorganisms, for example Streptococcus pyogenes and Mycoplasaates. The compounds of formula (1) are also effective against certain gram-negative bacteria, for example Pasteurella sp. In addition, the compounds produced by the method of the invention are well absorbed upon administration and provide good levels of antibiotic in the blood. For this reason, they can be used for the prevention and treatment of microbial infections in warm-blooded animals. For this purpose, a chemotherapeutically effective amount of a compound of formula I is administered to a parenterally or orally infected or endangered warm-blooded animal. In some cases, it is also possible to inject the compounds, for example in the form of a dust, into an enclosure in which, for example, animals or poultry are kept. These animals or poultry then inhale the dust containing the active substance present in the air, moreover, the powder may also enter the body through the eye mucosa, thereby treating or protecting the animals from infection.

The dose required to control the infection varies depending on the severity of the infection and the age, weight and general condition of the animal. The total dose for parenteral administration is usually in the range of 1 to 100 mg / kg. When administered orally, this dose is usually taken. it is in the range of up to 300 mg / kg, preferably 1 to 100 mg / kg.

Antibiotics obtained by the method of the invention may be formulated for the treatment of infections caused by bacteria and mycoplasmas. These veterinary compositions comprise as active ingredient a macrolide antibiotic of formula (I), a pharmaceutically acceptable salt or acyl ester thereof, together with one or more physiologically acceptable carriers or diluents.

Often, the most preferred route of administration of these compounds is the incorporation into feed or drinking water. The compounds can be incorporated into a variety of feeds, including dry feed, liquid feed and pelleted feed.

The method of incorporating veterinary drugs into feed is well known. The most preferred method is to produce a concentrated pre-feed which is then added to the feed. A typical pre-feed contains 1 to 400 g of active ingredient per kg and may be in liquid or solid form.

The resulting livestock feed or poultry will depend on the amount of antibiotic to be administered. Conventional methods are used to produce and pellet these feeds.

Injectable compositions containing the compounds of the present invention may be in the form of a suspension or solution. In the preparation of these compositions, it is understood that acid addition salts have better solubility in water than free materials. The free compounds of formula I are more soluble in dilute acids or acidic solutions than in neutral or basic solutions.

The compounds produced by the process of the invention are dissolved in physiologically acceptable compositions. These may be suitable solvents, preservatives such as benzyl alcohol and buffers. A useful solvent is, for example, water, a mixture of water and alcohols, 1-glycols and esters, for example diethyl carbonate. Aqueous solutions usually contain at most 50% by volume organic solvent.

Suspensions for injection are obtained using a liquid medium, optionally in the presence of an adjuvant. The medium may be, for example, aqueous polyvinylpyrrolidone, inert oils such as vegetable oils or highly purified mineral oils or aqueous carboxymethylcellulose.

Physiologically acceptable adjuvants are added to keep the compounds in suspension. These include, for example, a thickening agent such as carboxymethylcelluloses, polyvinylpyrrolidone, gelatin and alginates. Wetting agents may also be used as suspending aids. Other suitable suspending agents include lecithin, alkylphenol and polyethylene oxide addition products, naphthalene sulfonates, alkylbenzene sulfonates, and polyoxyethylene sorbitan esters.

A variety of substances affecting the hydrophilicity, density, and surface tension of liquid suspensions may help in individual cases to form a suspension suitable for injection use. For example, antifoams such as silicones, sorbitol and sugars may be used.

The invention will be illustrated by the following examples.

He did

20-deoxo-20- (N-benzylamino) tylosin g tylosin, 7.9 g benzylamine hydrochloride and 1.4 g sodium cyanoborohydride are dissolved in 50 ml methanol and the mixture is reacted under nitrogen atmosphere for 5 hours at room temperature. After completion of the reaction, the reaction mixture was poured into 600 ml of cold, saturated aqueous sodium bicarbonate solution and then extracted three times with 100 ml of chloroform. The resulting chloroform layer was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography eluting with chloroform: methanol: concentrated ammonium hydroxide (15: 1: 0.5) to give 3.8 g (71%) of a yellow product.

Elemental analysis for 6 molecular weight 1,006:

calculated:

found:

63.22% C, 64.01% C,

H 8.55, H 8.58,

N 2.78, N 2.69,

25.45 # O; 24.72 »O.

Melting point: 103.5 to 107 ° C

Specific rotation [.alpha.] D @ ²⁰ : -55.8 DEG (c 1, methanol)

Spectrum in ultraviolet light

LCH-max max

283 nm (S 22,900)

Mass spectrum (a / e) 673, 654, 175, 174, 145, 106, 91.

In a similar manner, the compounds of the following examples can be obtained

Example 2

20-deoxo-20- (K, N-dimethylamino) tylosin

119.2-120.3 ° C

Specific rotation at -40.4 ° (c 1, methanol)

Ultraviolet spectrum .HeOH (0- 282.5 nm (12500)

Mass spectrum (m / e): 944, 700, 770, 419, 175.

NMR (.delta. (Ppm)):

1.78 (and, 2.15 <8, C ₂₀ -N (Me) ₂ ) 2.52 (. ·, O ₃ -N (Me) _a ) 3.43 (E, C ₂ ,,, - 0lle), 3.60 (8, C ₃ ,,, - 0Me), *, 23 CD, H, «), 4.52 CD, »») « 4.92 (bt ' ^H 15 ^' 5,05 (d, H ,,,), 5.90 CD, h, ₃ ), 6.26 CD, ^ 0 ^ 7.33 CD, H ,,)>

Yield: 84%.

At 1. a d 3

20-deoxo-20- (N-anilino) tyloain

Melting point: 111-113 ° C

Optical rotation: M -86.0 ° (c 1, methanol)

Haot spectrum (m / e): 831, 658,190,175,145.

NMR spectrum (ppm):

1.77 (with, ^ 22 ^ ^- 2.46 (·, C ₃ -N (Me) g) 3.46 (E, C ₂ ,,, - Cbe), 3.56 (with, C ₃ ,,, - 0Ke), 4.23 CD, H, '), 4.54 CD, H ,,,,), 5.04 CD, ,,), 4.97 (bt, - ^H 15 ^}

AND

5.84 (bd. ^H , 3 ⁾ 6.23 (d, ^H 10 ⁾ ' 6.57 (d, o-H-anilino) 6.67 (d, o-H-anilino) 7.10 (d, m-H-anilino) 7.23 (d, H ,,).

Yield: 48%.

Example 4

20-deoxo-20- [N- (o-ethoxycarbonyl) anilino] tylosin Melting point: 122.5 to 124 ° C UV spectrum and MeOH ^X aax ^{(C): 282.3} "" ⁽² '^300> Yield: 34%.

Example 5

20-deoxo-20- (N-methylanilino) tylosin

Melting point: 106.5-109 ° C

Mass spectrum (m / e): Γ 006, 862, 688, 481, 175, 145 HMK spectrum, δ (ppm):

1.79 (with, Hgg), 2.40 (with, Cy-N (Me) ₂ ) 2.87 (with, C ₂₀ -N-Ue), 3.46 (with, C ₂ ,,, - 0Me), 3.59 (with, C ₃ ,,, - 0Me), 4.27 (d, H, '>, 4.54 (d, «,,,,,) 5,05 (d, «,,,>, 5.0 (b, h, ₅ ), 5.83 (bd , h, ₃ ), 6.21 (d, H, _o ), 6.7 (d, C ₂₀ -N- (oH (gH) Ph), 7.27 (d, ». 7.15 (d, C ₂₀ -N- (bH) PH).

Yield: 34%.

Example

20-deoxo-20- (N, N-dibenzylamino) tylosin MMR spectrum <5 (ppm):

1.78 (s, Hgg),

2.46 (s, _C3, -N (Me) _2), i.e.

3.46 (s, _C2 ,,, - 0Me)

3.60 (s, C ₃ , - 0 Me),

4.57 (d, H ,,,,),

5.88 (bd, H _13);

6.23 (d, H, _o)

7.3 (benzylic protons).

Yield: 70%.

Example 7

20-deoxo-20- (N-adamantylamino) tylosin

Melting point 108.5-10.5 ° C

Optical rotation: [α] -32.9 ° (o = 1, methanol)

Absorption spectrum in ultraviolet light:

, UeOH ^Z max 284 nm (9,800)

Mass spectrum (m / e): 1050, 732, 718, 543, 175, 157.

Yield: 82%.

Example

20-deoxo-20- (N-cyclohexylamino) tylosin

Melting point: 111-114 ° C.

Optical rotation: [α] -48.9 ° (c1, methanol)

Absorption spectrum in ultraviolet light:

UeOH max (£): 284 nm (20,000)

Mass Spec (m / e): 832, 664, 473, 175, 157, 145.

Yield: 50%.

Example 9

20-deoxo-20- (N-benzylamino) demycarosyltylosin g 20-deoxo-20- (N-benzylamino) tylosin was dissolved in 30 ml of 0.1 K hydrochloric acid and allowed to react at room temperature for 19 hours. After completion of the reaction, the reaction mixture was washed with 30 ml of chloroform and the resulting aqueous layer was adjusted to pH 8.0 with 1 N sodium hydroxide. The solution was extracted three times with 100 ml of chloroform, the chloroform layer was dried over anhydrous sodium sulfate and evaporated. pressure to dryness to give 820 mg of pale yellow crude material. This material was purified by silica gel column chromatography using a 20: 1: 0.95 mixture of chloroform, methanol and concentrated ammonium hydroxide as eluent. to give 620 mg of the title compound as a white solid, yield 72%.

Elemental analysis for (MW 862):

calculated:

found:

64.10% C, 64.04% C,

H 8.60, H 8.58,

N, 3.20, N, 3.25,

24.10% 0; 24.13% 0.

Mp 88.5-91.5 ° C.

Optical rotation: [α] = -35.2 ° (c, = 1, methanol)

Spectrum in ultraviolet light:

, CH, OH max ^{= 264 (C 22,000)} ·

The absorption spectrum IR (KBr): 3440, 2980, 2950, 1710, 1680, 1590, 1460, 1350, 1165, 1090 cm ^- '.

Mass spectrum (m / e): 756, 672, 565, 482, 174, 91.

In a similar manner, the compound of the following example can be obtained.

He did

0-deoxo-2 0- (N-thylanil.no) demykaro syltylo sin

Melting point: 95.5 to 98.5 ° C.

Absorption spectrum in ultraviolet light:

MeOH max (C):

283 nm (20,600)

Mass spectrum (m / e): 862,688,190,174. NMR spectrum (δ (ppm)):

1, 77 (with, h ₂₂ ), 2.49 (with, C ₃ -N (Me) ₂ 2.87 (with, C ₂₀ -N-Me), 3 <46 (with, C ₂ ,, - OMe>, 3.59 (with, C ₃ , - C e, 4.33 CD, H, '), 4,56 (and, H, '), 5.00 (dt, 5.84 (bd, , h ₁₃ ), 6.63 CD, ^H 10 ⁾ > 6.70 (and, o-H-aniline) 6.73 Ca, p-H-aniline) 7.20 CD, m-H-aniline) 7.2 CH ,, ).

Yield: 43 »

Example 11

20-deoxo-20- (N-morpholine) tylosin

A solution of 500 mg of tylosin, 480 mg of morpholine and 30 mg of sodium cyanoborohydride in 5 ml of methanol was allowed to stand at room temperature under a nitrogen atmosphere for 24 hours. After completion of the reaction, the reaction mixture was poured into 50 mL of aqueous saturated sodium bicarbonate solution and then extracted three times with 80 mL of chloroform. The resulting chloroform layer was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure to give the title compound as a crude product. This material was purified by silica gel column chromatography using a 30: 1: 0.05 mixture of chloroform, methanol and concentrated ammonium hydroxide to give the title compound as a white solid (430 mg, 80%).

Elemental analysis for ^ 50 ^ 6 ^ 2 ^ 17 (MW 986):

calculated; Found: C, 60.85; H, 8.72;

2.83% N, 27.62% 0.

2.84% N, 27.59 »O.

Melting point: 122.5 to 124.5 ° C.

Optical rotation: [α] = -39.6 ° (c = 1, methahnol)

Absorption spectrum in ultraviolet light:

CH, 0H ^A max ^{283.5 (L} '9500)

In a conventional manner, the compound of the following example can be obtained:

Example 12

20-deoxo-20- (4-N-methylpiperasinylamino) tylosin

Melting point: 95.0 to 98.0 ° C

Optical rotation: -55.4 ° (c 1, methanol)

Absorption spectrum in ultraviolet light:

&Gt; MeOH (t) 283 nm (17,700) n &

Mass spectrum (m / e): 1 014, 870, 680, 175, 157, 145

HMR-spectrum δ (ppm);

1.79 (with, «22), 2.31 (E, 4-Me) 2.47 (with, C ₃ : -N (Me) ₂ ) 3.47 (E, Og 'OMe') 3.60 (with, C ₃ ,,, - 0Me), 4.27 <4, 4,56 <a. H ,,,,), 5,07 (and, H ,,,),

4.95 (b, H ₁₅ ), 5.86 (d, H _{) 3} ),

6.26 (d, _{H) 0)}

7.27 (d, H +).

Yield: 69%.

The antibacterial activity of typical compounds of formula (1) is shown in the following table, which shows the results of experiments with the determination of minimum inhibitory concentration (HIC) in a standard agar solution, cultured for 24 hours at 37 ° C.

Table

Minimum inhibitory concentration (jug / ml) Example number ...... 9 , 0. Staphylococcus aureus PDA 209P 0.4 0.4 Bacillus subtilis PCI 219 0.4 0.4 Bacillus cereus IPC 300 0.4 <0.1 Micrococcus leuteus PCI 1001 <0.1 <0.1 Mycobacterium smegmatis ATCC 607 3.12 25 Esterichia coli NIHJ 25 > 100 Klebsiella pneumoniae PCI 602 50 50 Salmonella typhimuriua KB 20 50 > 100

Animal experiments have also shown that 20-deoxo-20- (N-benzylamino) demycarosyltylosin when administered orally to mice infected with Streptococcus pyogenes has an ED _{50 of} 39 mg / kg, i.e. almost twice that of demycarosyltylosin.

Claims (5)

SUBJECT OF THE INVENTION ,. A process for the preparation of 20-aminotylosine derivatives of the general formula I wherein H represents a hydrogen atom, a hydroxyl group or a mycinosyloxy group, Q represents a hydrogen atom, X is a radical of formula wherein (1) r'O and r '' are independently hydrogen, C 1 -C 4 alkyl or C 3 -C 10 cycloalkyl, or formula - < ^CH 2) _n -Ph wherein U is an integer of 0, 1 or 2a Ph represents a phenyl radical optionally substituted by a C 1 -C 4 alkoxycarbonyl, amino, halogen, C 1 -C 4 alkyl or alkoxy radical, (ii) a single substituent R c or r represents a group of the formula wherein, 12 ( iii) r 10a represents a hydrogen atom or a C 1 -C 4 alkyl radical, the latter as defined in (i) or (11), together with the nitrogen atoms to which they are attached, forms a 5- to 7-heteroatomic ring, the heteroatom is an alkali or a dical, as well as pharmaceutically acceptable salts and acyl esters, characterized in that a) cleaving off the mycarose sugar from an aacrolide of formula I corresponding to formula I wherein Q is replaced by Q * and Q * represents an acarose by acid hydrolysis to give a compound of formula I wherein Q is hydrogen b) optionally, the product obtained in step (a) is esterified and / or converted into a salt. 2. A process according to claim 1, starting from a compound of formula (I ') to form an acrolide of formula (I) wherein R is mycinoayloxy and the other formulas are as defined above, or salts or an acyl ester thereof, acceptable. from a pharmaceutical point of view. 3. A process according to claim 1, wherein the compound of formula (I) is formed to form a macrolide antibiotic of formula (1) in which X is amino, dimethylano, aniline, N-ethylaniline, o-ethoxycarbonylaniline. , benzylamino, aainobenzylaainoacin, halobenzylamino, cyclohexylaminoacin or an aphoroline group, or a pharmaceutically acceptable salt or acyl ester thereof. 2353 * 3 4. A process according to claim 1, starting from a compound of formula I 'to give 20-deoxo-20- (N-benzylamino) demycarosyltylosin. 5. A process according to claim 1, starting from a compound of formula I * to give 20-deoxo-20- (N-morpholino) demycarosyltylosin.