HU195820B - Process for producing new 6-alkyl-analogues of spectinomycin and pharmaceutical compositions containing them as active components - Google Patents

Abstract

Field of the Invention The present invention relates to novel antibacterial action spectinomycin derivatives of the formula I wherein R 'is C 1 -C 12 alkyl or (C 4 -C 8) cycloalkyl (C 1 -C 4) alkyl. and pharmaceutically acceptable acid addition salts thereof and the preparation of hydrates thereof by chemical methods. R * 1 195820 -1-

Description

The present invention relates to a process for the preparation of novel 6 * - < g > The compounds of the present invention have excellent antibacterial activity.

Spectinomycin A is a known anitibiotic and was first prepared microbiologically (U.S. Patent No. 3,234,092).

Some analogs of spectinomycin are described in Rosenbrook, Jr., et al., J. Antibiotics, 28, 953-960 (1975) and ibid, 31, 451 (1978), and Carney et al., Ibid, 30, 960 (1977). The chloro-deoxy derivatives of spectinomycin are described in U.S. Pat. Foley et al., J. Org. Chem. 43 (22). 4355-4359 (1978) describes 9-epi-4- (R) -dihydro-spectinomycin. However, neither of the mentioned spectinomycin analogs or derivatives mentions biological efficacy in the mentioned publications.

Lemieux a Can. J. Chem., 51, 53 (1973) that the 5-hydroxy group of 2-deoxy-streptamine (1) reacts preferentially with the tri-O-acetyl-2-deoxy-2-nitroso (2). alpha-D-glucopyranosyl chloride to give an alpha-pseudodisaccharide for which CBz represents a benzyloxycarbonyl group. Mallams et al., J. Chem. Soc. Perkin I, 1118 (1976) utilize the Lemieux method for the synthesis of diis trisaccharides.

Oxime degradation is suggested by Lemieux et al. And Mallame et al. In their publications mentioned in the previous paragraph.

In 1979, Hannesaian et al. Described the chemical synthesis of spectinomycin.

White et al., July, 1979, in Tetrahedron Letters, describe chemical synthesis for the production of spectinomycin and analogues thereof.

We have discovered that a wide variety of 6'-alkyl spectinomycin analogs can be prepared from dienes by rose-catalyzed Grignard addition using various intermediates useful in the Reaction Batch. In the reaction sequence (enon acylates), the C-6 'conversion takes place.

Accordingly, the present invention relates to a process for the preparation of compounds of formula I from compounds of formula IV. Compounds of formula (IV) may be prepared from enone acylates of formula (VII).

The process of the invention is described with reference to Scheme B. In Reaction Scheme B

R 'is alkyl of 1 to 12 carbon atoms or cycloalkyl of 4 to 8 carbon atoms; Containing 1-4 carbon atoms (alkyl,

R'j and R'e as protecting groups are aralkoxycarbonyl, halogenated alkoxycarbonyl or alkoxycarbonyl; and

Rio represents an acyl group.

The carbon numbering used in Formula (I) is referred to throughout this specification, although this numbering is different from the usual.

The compounds of the present invention include acid addition salts and hydrated forms of the compounds of formula (I).

Examples of alkyl groups mentioned in the substituent reports include the methyl, ethyl, propyl, butyl, pentyl, hexyl, heptylene octyl and isomeric forms thereof.

Examples of the acyl group include formyl, acetyl, propionyl, butyryl and pentanoyl.

Examples of the halogenated alkoxycarbonyl include mono-, di- or trihaloxymethoxycarbonyl-, mono-, di- or trihalogenethoxycarbonyl-, mono- or trihalo- propoxycarbonyl-, mono-, di- or trihalogen-. butoxycarbonyl or mono-, di- or trihalogenopentoxycarbonyl, or isomeric forms thereof.

Halogen atoms include fluorine, chlorine, bromine and iodine.

The term "aralkoxycarbonyl" refers to benzyloxycarbonyl, phenylethoxycarbonyl, phenylpropoxycarbonyl or diphenyloctyloxycarbonyl, and isomeric forms thereof, and fluorenylmethoxycarbonyl.

By alkoxycarbonyl is meant isopropoxycarbonyl, tert-butoxycarbonyl or tert-pentoxycarbonyl.

It is further noted that if more than one hydroxy or alkoxy group is present in the sugar molecules mentioned herein and in the claims, they may be the same or different.

The compounds of formulas II, III and IV are novel compounds. The dienones of formula IV are very versatile, so-called Michael acceptors.

The "alpha-anomer" refers to the anomer carrying a substituent at the 1'-position below the plane of the ring system, while the "beta-anomer" to the carbon at the 1'-position is an anomer having the same configuration as spectinomycin.

Among the compounds obtainable by the process of the present invention, the beta-anomers of the compounds of formula (I) have the desired biological activity. This glycosidic configuration can be found in the spectinomycin shown in Scheme B.

In Scheme B, Step 1, an enone acyl ethanol of formula (VIII) is reacted with dimethylforminide dimethylacetate in a solvent to give an enamine (VI). The reaction may be carried out at a temperature between 25 ° C and the reflux temperature of the reaction mixture, preferably at 40-80 ° C. The reaction time is from 1 to 48 hours, preferably from 2 to 10 hours. Dimethylformamide dimethylacetal is used in excess. The preferred reaction time is 7 hours and the preferred solvent is dimethylformamide. Other acetals of dimethylformamide, such as ditert-butyl acetal, may also be used.

The starting enone acetate (VII) and methods for its preparation are described in U.S. Patent 4,351,771.

The enamines of formula (VI) may be prepared from the reaction mixture by conventional methods such as extraction, chromatography or a combination of such methods.

In step 2, the side chain of the enamines of formula (VI) is reduced by the silver cyanoborohydride method to give an amine of formula (V). The latter can then be prepared from the reaction mixture by conventional means such as extraction, chromatography or a combination of such methods.

In step 3, an amine of formula (V) is reacted with an alkyl halide in the presence of a solvent to give a dienone of formula (IV). The reaction may be carried out at a temperature of 0 'C to 100' C for a period of 1 to 30 hours, for example using excess methyl iodide. Preferred reaction temperatures and times are 20-40 'C and 1-20 hours. Suitable solvents are, for example, methylene chloride, chloroform, tetrahydrofuran or diethyl ether. The preferred solvent is methylene chloride.

An alternative and preferred method for carrying out Step 3 is to react an amine of formula (V) with 3-chloroperbenzoic acid in a solvent to give a dienone of formula (IV). The reaction is carried out at a temperature between -20 'C and 50' C for 1-60 minutes. The preferred reaction temperature and reaction time are 20-30 ° C and 5-20 minutes. Suitable solvents are, for example, a mixture of ethyl acetate and Skellysolve B, tetrahydrofurene, dioxane or methylene chloride. The preferred solvent is the first mentioned mixture.

In step 4, a dienone of formula (IV) is reacted with a Grignard reagent in the presence of a copper catalyst to give a compound of general formula (III). This reaction is carried out in a solvent at a temperature of -130 ° C to 0 ° C for 1 minute to 2 hours. Preferred reaction temperatures and reaction times are -78 ° C to -50 ° C and 1-10 minutes. Examples of the solvent include diethyl ether, 1,2-dimethylethane, tetrahydrofuran or diisopropyl ether. The preferred solvent is tetrahydrofuran.

In step 5, an enone of formula (III) is reduced to give a protected 6-alkyl spectinomycin of formula (II). This step is extremely important for carrying out the process of the invention for a number of reasons. It is extremely difficult to reduce the 4 ', 5'-double bond without reducing the 3'-carbonyl group. As mentioned above, palladium catalysts, such as palladium catalysts containing 10% metal barium sulfate, can be used with partial success, but protecting groups, especially benzyloxycarbonyl groups, are also cleaved, which further increases the separation problems. In addition, palladium catalysts are ineffective with analogs containing polysubstituted sugar moieties. Therefore, the catalyst is preferably platinum oxide in the presence of a solvent and a base.

Another major advantage of using the latter catalyst is that the protecting groups remain intact during the reduction so that the products can be purified by chromatography before being subjected to one of the many protecting group deprotection methods. The reaction is carried out by dissolving an enone of formula (III) in a solvent, adding the base and platinum oxide to the resulting solution and treating the resulting mixture with hydrogen gas. The reduction is carried out at 20-40 ° C for a reaction time of 1-4 hours. Preferred reaction temperatures and reaction times are 20-40 ° C and 2-3 hours.

An alternative and preferred method for performing Step 5 is the reduction of a didehydro spectinomycin of formula (III) to a protected 6'-alkyl spectinomycin of formula (II) with a solution of lithium tri-sec-butylborohydride. The reactants are carried out by dissolving a didehydro-spectinomycin of formula (III) in a solvent, adding a solution of the lithium-containing reagent, followed by a base and water. The reduction is carried out at a temperature of -130 ° C to -10 ° C for a reaction time of 5-63 minutes. Preferred reaction temperatures and reaction times are -73 'C to -10' C and 5-40 minutes. Suitable solvents are, for example, tetrahydrofuran, dioxane, 1,2-dimethoxyethane or diethyl ether. The preferred solvent is tetrahydrofuran.

In step 6, a compound of formula II is deprotected to give a final product of formula I. The conditions for the deprotection operation depend on the particular protecting groups used, i. E. On the actinin ring, on the airinocBoportol protecting groups R'j and R'e. When the protecting group is benzyloxycarbonyl or aralkoxycarbonyl, the protecting group is 0.304. 10 ⁵ Pa and 1,418. At a hydrogen pressure of 10 Pa, in the presence of a conventional catalyst such as palladium carbon, palladium on carbon or palladium on barium sulfate, tetrahydrofuran in a solvent such as isopropanol, absolute ethanol, methanol, ethyl acetate, toluene. In some cases, it is beneficial _; if hydrogen is generated in situ from methanol-hang y-acid solution.

Alternatively, the compounds of formula (II) having R'1 and R'e as alkoxycarbonyl or aryloxycarbonyl as protecting groups may be deprotected with an acid in a solvent such as nitromethane or methylene chloride.

When R'i and R'e are haloalkoxycarbonyl, the deprotection is preferably carried out with zinc.

A particularly preferred reaction sequence for the preparation of compounds of Formula VI is shown in Scheme C.

In Scheme C, Step 1, N, N'-di (benzyloxycarbonyl) -spectinomycin of formula (X) (prepared by the Journal of the American Chemical Society, 85, 2652 (1963)) is reacted with formic acid, ethyl acetate. in the presence of acetate and pyridine. The reaction is carried out at a temperature between -70 'C and + 10' C for 0.5 to 30 hours in the presence of a solvent. The preferred reaction temperature is between -40 ° C and 0 ° C and 10-18 hours. Suitable solvents are, for example, ethyl acetate, tetrahydrofuran, methylene chloride, chloroform or 1,2-dimethylethane. The preferred solvent is ethyl acetate. In step 1, the corresponding compound of formula IX is formed.

In step 2, the compound of formula (IX) is reacted with acetic anhydride (Rio) aO and a base to give the corresponding compound of formula (VIII). The reaction may optionally be carried out in the presence of a solvent. The reaction is carried out at C, preferably 50-70 ° C, for a reaction time of 2 to 30 hours, preferably 10 to 20 hours. Suitable solvents are, for example, ethyl acetate, tetrahydrofuran, 1,2-dimethoxyethane or chloroform. The preferred solvent is ethyl acetate.

In step 3, the compound of formula VIII is subjected to light treatment in the presence of dibromodimethylhydantoin to form a carbonyl group at the 3-position carbon atom as shown in formula VII. The reaction is carried out at a reaction time of 10-100'C, preferably 40-80'C, with a reaction time of 2-60 minutes, preferably 10-20 minutes. As a result of this step, a double bond is formed at the 4,5-position.

In Step 4, the compound of general formula (VII) is reacted with diraethylformamide acetate in the presence of dimethylformamide and then the resulting intermediate is reacted with methanol or water to give the product of general formula (VI). Step 4 is carried out at a reaction temperature of 30-90 ° C, preferably 40-70 ° C, for 2-6 hours, preferably 3-5 hours.

The intermediates obtained according to each of the previous steps, if desired, are the reaction. may be isolated from the mixture in a known manner.

Any of the known methods for isolating the compounds of formula (I) may also be used. If the isolation is carried out under anhydrous conditions, the compounds of formula (I) having a carbonyl group at the 3'-position are obtained. Conversely, isolation in the presence of water yields compounds hydrated at the 3'-position, except for compounds of formulas VII and VI, which are unable to hydrate.

The crude end products can be purified by adsorption chromatography on a column of weakly acidic ion exchange resin such as Amberlite IRC-50 or CG-50, eluting with water, methanol, ethanol, diethyl ether, tetrahydrofuran, 1,2-dimethoxy, using p-Cioxane. The solvents listed above include hydrochloric acid, hydrobromonide, hydrogen iodide or sulfuric acid as the acid.

Acid addition salts may be prepared by acidifying the compounds of formula (I) to a suitable SBwal below pH 7.0, preferably at pH 2-3. Suitable acids for this purpose include, for example, hydrochloric acid, sulfuric acid, phosphoric acid, sulfuric acid or hydrobromic acid.

The acid addition salts and the cationic salts can be used for the same biological purposes as the free compounds of formula (I).

The compounds of the present invention inhibit the growth of microorganisms under various conditions, such as the beta-configurated compounds of formula (I) being active against Escherichia coli and thus useful in reducing or preventing the formation of sludge formed by this microorganism in papermaking systems. These beta anomers can also be used to extend the lifespan of cultures of Trichomonas fetus, Trichomonas hominis, and Trichomonas vaginalis by depleting these cultures of Escherichia coli contamination. These beta-anomers are also active against Bacillus subtilis and can thus be used in fish or fish products to minimize or eliminate the odor caused by the microorganism. These beta-anomers can also be used to treat laboratory desks and racks used in glass washes and mycology laboratory equipment. Beta anomers are also effective against Klebsiella pneumoniae.

The compounds of formula (I) are also effective for the treatment of bacterial infections, e.g., 49-dul gonorrhea in mammals, including man.

Particularly good antibacterial activity is shown by the compounds of formula I wherein R 'is a straight, branched or cyclic alkyl group and the longest moiety contains from 1 to 4 carbon atoms in the branched or cyclic chains. Examples of such R 'substituents include ethyl, propyl, isopropyl, butyl, tert-butyl or cyclohexyl. 6 '- (n-Propyl) Spectinomycin, 6' - (n-Butyl) Spectinomycin or 6 '- (n-Pentyl) Spectinomycin are particularly effective antibacterial compounds.

The compounds of the present invention, when mixed with excipients and / or excipients commonly used in the pharmaceutical art, can be formulated into pharmaceutical compositions suitable for administration to humans and animals, such as sterile parenteral solutions or suspensions, eye drops and water-in-oil emulsions.

For parenteral administration, liquid preparations are prepared using the active ingredient and a sterile diluent, preferably water. Depending on the nature and concentration of the diluent employed, the active ingredient is suspended or dissolved in the diluent. In preparing solutions, the active ingredient is dissolved in water for injection and then sterilized by filtration before filling into a suitable vial or ampoule and sealing with one of the latter. Additives such as topical anesthetics, preservatives and buffering agents are preferably dissolved in the diluent. To increase stability, the resulting composition is frozen after filling into a vial and the water is removed under vacuum. The dry lyophilized powder is then sealed in the vial and a vial of water for injection is added to the pharmaceutical package to restore the liquid to a state prior to use. Parenteral suspensions may be prepared in the same manner except that the active ingredient is suspended in the diluent instead of being reconstituted and sterilization is not performed by filtration. In this case, the active ingredient is sterilized with ethylene oxide before being suspended in the sterile diluent. Preferably, a surfactant or wetting agent is added to the composition to ensure a uniform distribution of the active ingredient.

The term "dosage unit" as used herein refers to physically discrete units suitable for administration of a uniform amount of active ingredient to humans and animals. Each such dosage unit form contains an amount of active ingredient calculated on the basis of the desired therapeutic effect, together with such carrier and / or excipient. Examples of such dosage units are ampoules, vials, aerosol formulations for measuring a measurable quantity of refrigerant, kits comprising any one of such formulations, and any other form known in the art.

Effective amounts of the active ingredient are, of course, used in the treatment. The magnitude of these depends on a variety of factors well known to those skilled in the art, such as the mode of administration and the potency of the compound. In humans, the single dose may be about 2-4,000 mg for the treatment of bacterial infections. In particular, a single dose may be, for example, about 5 mg to about 200 mg.

The present invention is illustrated by the following reference examples and embodiments. It will be appreciated by those skilled in the art that the corresponding stereoisomers of the compounds mentioned in these examples are also encompassed within the scope of the invention.

Reference Example 1

N, N'-di (Benzyloxycarbonyl) -2,6-di-O-formylcytinomycin

To a 5 L flask equipped with a stirrer and a thermometer, 1.33 L of ethyl acetate and 255 mL of 97% formic acid are added, followed by the slow addition of acetic anhydride (618 mL) while cooling in a dry ice / acetone cooling bath. stay below. Pyridine (1.09 L) was then added slowly to the reaction mixture, maintaining the temperature at -40 ° C. Subsequently, 100 g of Ν, Ν'-dibenzyloxy-spectinomycin are added and the cooling bath is allowed to warm to room temperature with stirring overnight. The reaction mixture was then diluted with ethyl acetate (1.65 L), quenched with 0.5 N brine (675-675 mL), twice with water (675-675 mL), 10% aqueous sodium bicarbonate (675-675 mL), and finally. It is also washed twice with 675-675 ml of saturated aqueous sodium chloride solution. After drying and evaporation of the solvent, 105 g of the title compound are obtained.

CMR (De-acetone) t 201.6, 161.4, 161.0,

157.1, 133.6, 129.1,

128.4, 128.1, 97.1, 92.1,

73.7, 71.7, 68.3, 67.7,

67.6, 65.9 and 56.1 ppm.

-511

Reference Example 2

Reference Example 4

N, N'-Di (benzyloxycarbonyl) -2,6-di-0-formyl-2 '- (0-acetyl) -epektinomicin enol acetate

Weigh 7.0 g into a 250 ml flask

N, N'-di (benzyloxycarbonyl) -2,6-di-O-formyl-epectinomycin, 15.0 ml acetic anhydride, 30.0 ml pyridine, 0.5 g dimethylaminopyridine and 50 ml ethyl acetate (the other components are added to the latter) and the resulting reaction mixture is kept at 55 ° C overnight. The reaction mixture was then diluted with ethyl acetate (500 mL) and hexane (500 mL), followed by 3 x 1 L 0.6 N hydrochloric acid, 0.5 L 0.6 N hydrochloric acid (1 x 500 mL), and saturated aqueous sodium bicarbonate (500 mL). and twice with 500 ml of saturated aqueous sodium chloride solution. The organic layers were combined, dried over sodium sulfate and evaporated to give 7.0 of the title compound.

CMR (De-acetone): 168.9,

161.0,

137.8,

161.4, 139.9,

128.4,

167.7,

157.1,

129.1,

128.2, 128.1, 122.8, 94.7,

92.2, 74.1, 71.3, 69.1,

67.6, 67.2, 56.3, 55.5,

31.4, 30.7, 21.1, 21.0, 20.7 ppm.

Reference Example 3

N, N'-Di (Benzyloxycarbonyl) -2,6-di-O-formyl-2 '- (0-acetyl) -4', 5'-didehydro-spectinomycin was dissolved in ml of ethan tetrachloride. g of N, N'-di (benzyloxycarbonyl) -2,6-di-O-formyl-2 '- {0-acetyl) -spectinomycin-enol acetate and then 1.05 g of dibromo dimethyl hydantoin is added. Using the visible light as the initiator, the resulting reaction mixture is refluxed. After 20 minutes, the resulting solution was cooled and diluted with chloroform (200 mL) and water (100 mL). The phases are separated35

N, N'-di (Benailoxycarbonyl) -2 '- (0-acetyl) -6 - [(dimethylamino) methyl] -4', 5'-didehydro-spectinomycin

22.0 g (34.4 mmol) of N, N'-di (benzyloxycarbonyl) -2 '- (0-acetyl) -4' ^, 5,5-didehydro-spectinomycin and 100 ml of dimethylformamide dimethylacetal A solution of 100 ml of dimethylformamide was heated at 50-55 ° C for 7 hours under Drier protection. The resulting brown solution is evaporated in vacuo and the residue is chromatographed on 250 g of silica gel which is wet filled in a 2 liter sintered glass funnel. Elution was carried out over 1.5 hours with 2 L of 5% acetonitrile in chloroform, 3 L of 33% acetonitrile in chloroform and 6 L of 50% acetonitrile in chloroform, 0.5 L fractions. The fractions were evaluated by thin layer chromatography (eluent: acetonitrile / chloroform = 1: 1 by volume) and the thermally containing fractions were poured. This gives 19.78 g of pure product on the one hand and 3.30 g of product containing a certain amount of dimethylformamide on the other hand. The latter portion was subjected to repeated chromatography on 60 g of silica gel to give 2.5 g of pure product. Total yield 22.53 g (94% yield). Circular Dichroism - CD (Methanol):

(Θ) 311 E · - 10,400 ± 1,200.

(3) 265 na = 2500 ± 1200.

(3) 246 na - 1200 ± 1200.

IR spectrum (bubble):

PMR (CDCb, á):

3380, 1750, 1695, 1675, (shoulder), 1600, 1555, 1500, 1385, 1350, 1280, 1240, 1195, 1185, 1145, 1110, 1085, 1065, 1025, 1000, 960, 770, 740, and 645 cm ¹ .

2.13 (3H, s), 2.82 (3H,

s), 2.89 (6H, broad s),

2.91 (3H, s), 5.10 (4H, broad s), 5.16 (1H, s), 5.97 (1H, s) and 7.30

they are apart, then the organic phase 60 ml (10H, b) aqueous sodium chloride -oldattal Moes, nitrate CMR (CDaCOCDa): 20.9, 31.2, 31.5, 30-44 dried over um sulfo evaporate, (wide), 36.3, 59.6, 5.65 g the title ve ₅₅ 66.2, 67.3, 74.6, 75.4, at stake. 88.7, 94.9, 95.0, 95.1, CMR (Ds-acetone): 182.4, 173.4, 170.0, 95.5, 128.3, 129.1, 137.9, 161.6, 161.0, 157.0, 138.0, , 150.3, 157.2, 137.6, 129.2, 128.6, 157.7, , 163.3, 169.8,

128.2, 103.4, 95.7, 93.1, 73.7, 72.1, 67.8, 66.9, 56.1, 55.7, 31.8, 30.6 and 21, 0 ppm.

171.2 and 180.3 ppm.

Mass Spectrum, m / c (diTMS): 839 (M +), 824, 797, 779, 730 and 688 (peaks aligned).

-613

Calculated molecular weight: 839, 3480

Molecular Weight: 839, 3466

Reference Example 5

N, N'-Di (benzyloxycarbonyl) -2 '- (0-acetyl) -6 - [(dimethylamino) methylene] -4', 5'didehidro-Bpektinomicin

Dimethylformamide (39.0 mL) was dissolved in N, N'-di (benzyloxycarbonyl) -2,6-di-O-formyl-2 '- (O-acetyl) -4', 5 '(5.9 g). -dehydro-spectinomycin followed by addition of 36.0 ml of dimethylformamide dimethylacetal. The resulting reaction mixture was heated at 55-60 ° C for 4 hours, then 36 'ml of methanol was added and heating was continued at 55-60 ° C for 3 hours. The residue was taken up in chloroform (15 ml) and the resulting solution was applied to a silica gel filter (200 ml). The product was liberated from the residual dimethylformamide by sequential elution with 0.5 L chloroform, 0.8 L 5% acetonitrile in chloroform, 150 mL 30% acetonitrile in chloroform and 1 L acetonitrile. 2.71 g of the title compound are isolated from the appropriate fractions. CMR (De-acetone): 180.3, 171.2, 169.8,

157.0, 150.3, 137.9,

129.0, 128.8, 95.5, 95.2,

95.1, 94.8, 88.7, 75.3,

67.3, 66.2, 59.9, 65.7,

41.0, 31.4, 30.7, and 21.2 ppm.

Reference Example 6

N, N'-Di (benzyloxycarbonyl) -2 '- (0-acetyl) -6' - (dimethylamino-pyridylmethyl) -4 ', 5'-didehydro-spectinomycin

Dissolve 20.0 g (28.78 mmol) of N, N'-di (benzyloxycarbonyl) -2 '- (0-acetyl) -6' - (dimethylaminomethyl) -4-methanol in 200 ml of methanol ^. 5'-didehydro-Bpectinomycin and 5 mg methylarance, and the resulting solution was adjusted to pH 4.0 with 2N methanolic hydrogen chloride. Sodium cyanoborohydride is then added over a period of 20 minutes, with frequent pH adjustments, using 2N methanolic hydrochloric acid. After addition of a total of 2.0 g (31.83 mmol) of sodium cyanoborohydride, the resulting solution was stirred at room temperature for 1 hour. TLC (1: 9 methanol: chloroform) as eluent indicated the starting material to be converted. The solution was first concentrated and diluted with 500 ml of ethyl acetate, 250 ml of 0.1 N aqueous sodium hydroxide solution, 25 ml of 1.0 N aqueous sodium hydroxide solution and finally 250 ml of a saturated aqueous solution of sodium chloride. . After separation of the organic phase, the reaction mixture is washed with 200 ml and then 100 ml of a saturated aqueous sodium chloride solution. The three different aqueous phases are washed with 200-200 ml of ethyl acetate, and the organic phases are combined, dried over sodium sulfate and evaporated. This gave 17.93 g (89%) of the title compound as a foam.

CD (CIbOH): (Θ) μι ». = 12,000 ± 1100 (θ) 270 Β · - ± 1100

UV spectrum (C 2 H 5 OH): 273 nm (8800) and 377 nm (shoulder, 63).

PMR (CDCl 3, i): 2.13 (3H, s), 2.17 (6H,

s), 2.46 (3H, s), 3.05 (3H, s), 3.06 (3H, s),

5.05 (4H, d), 5.34 (1H, s), 5.89 (1H, s) and 7.26 (10H, d).

CMR (CD3COCD3): 20.9, 31.3, 45.0, 56.1,

57.1, 61.3, 66.1, 67.3, 74.0, 75.4, 93.7, 96.0,

103.1, 128.3, 129.1,

137.9, 157.2, 170.0,

174.9, ee 183.1 ppm.

6a. PREPARATION

N, N'-Di (benzyloxycarbonyl) -2 '- (0-acetyl) -6' - (dimethylamino) -4 ', 5'-didehydro-epektinomicin

57.0 g (78.7 mmol) of N, N'-di (benzyloxycarbonyl) -2 '- (0-acetyl) -6' - [(dimethylamino) methylene) -4 ', 5 ^, -didehydro-apectinomycin, 450. of ethyl acetate and 105 ml of methanol and the resulting mixture was adjusted to pH 4 with 2N hydrochloric acid. Then, 7.3 g of sodium cyanoborohydride are added over 5 minutes, with frequent pH adjustment using 2N hydrochloric acid. After the addition of sodium cyanoborohydride was completed, the reaction mixture was maintained at pH 4 for another 15 minutes and then subjected to thin layer chromatography using a 1: 9 by volume mixture of methanol and chloroform as eluent. The starting material was found to be transformed. The reaction mixture was then stirred with a solution of 175 g of sodium hydroxide in 350 ml of water and the layers were separated. The upper layer was washed with 250 mL of saturated aqueous sodium hydroxide solution and 150 mL of Skellysolve B was added to break the emulsion. The two aqueous layers were backwashed with ethyl acetate (300 mL) and Skellyeolve B (75 mL). The combined organic phases are dried over sodium sulfate and the sodium sulfate is filtered off to give a filtrate containing the title compound.

-715

Reference Example 7

N, N'-Di (benzyloxycarbonyl) -2 '- (0-acetyl) -6'metilén-4', 5'-didehydro-spectinomycin

Dissolve 17.43 g (25.0 mmol) of N, N'-di-benzyloxycarbonyl-2 'in a mixture of 200 ml of methylene chloride and 18.0 ml (41.04 g, 289 mmol) of methyl iodide. - (0-Acetyl) -6 '- (dimethylaminomethyl) -4', 5'-didehydro-spectinomycin and the resulting solution was stirred for 6 hours at Ezo temperature. The reaction mixture was allowed to stand at 10 ° C for 16 hours, then concentrated and dissolved in a 1: 9 by volume mixture of acetonitrile and chloroform. The resulting solution was chromatographed on 300 g of silica gel, eluting with 4 liters of said solvent mixture, followed by 4 liters of acetonitrile / chloroform (1: 3 by volume). The first eluting pure material was collected by pooling and evaporating the appropriate fractions to give the title compound (10.03 g, 61%).

CD (CH 2 OH): (O) 33. = 26 100 + 2300 (ö) j «o _n . = 44,800 + 2300 {0h «». = 10,800 ± 2300

UV spectrum (C 2 H 5 OH):

204 (shoulder, 22,500), 241 (6,750), 250 (shoulder, 5,900),

256 (shoulder, 4900) and 293 nm (14,650). PMR (CDCl3, 5): 2.13 (3H, a), 3.03 (3H, s), 3.06 (3H, s), 5.05 (4H, d), 5.44 (1H the), 5.61 (1H, q), 5.95 (LH, s), 6.11 (1H, s), 6.19 (LH, s) and 7.29 (10H, CMR (CD3COCD3): s ,. 20.9, 31.4, 57.2, 60.1, 66.2 ,, 67.4, 74.1, 74.7, 75.6, 94.0, 96.1, 104.3, 125.3, , 128.4, 129.2, 131, l · , 138.0, 138.1, 157.6 , 166.6, 170.2 and 184.0 ppm.

7a. PREPARATION

N, N'-di (Benzyloxycarbonyl) -2'- (0-acetyl) -6'methylene-4 ', 5'-didehydro-spectinomycin

6a. REFERENCE EXAMPLE N, N'-Di-Benzyloxycarbonyl-2 '- (O-acetyl) -6' - (dimethyl-ararinoethyl) -4 ' To the solution of 'didehydro-spectinomycin' is added 15 g of 3-chloroperbenzoic acid, and the reaction mixture is stirred for 10 minutes with the addition of 300 ml of a saturated aqueous solution of sodium bicarbonate. The reaction mixture thus obtained was separated into its phases and the organic phase was washed with 250 ml of a saturated sodium chloride solution and then methanol was added to break the emulsion. The two aqueous layers were washed with ethyl acetate (250 mL) followed by Skellysolve B (75 mL), the organic layers were combined, dried over sodium sulfate and evaporated. The resulting solid was dissolved in chloroform and chromatographed on 1 liter of silica funnel in a 1 L silica funnel, eluting successively with 4 L of chloroform, 2 L of 2% acetonitrile in chloroform, 2 L of 5% acetonitrile in 4 L of chloroform. 10% acetonitrile in chloroform and 4 liters 15% acetonitrile in chloroform. 10-14. fractions were combined and evaporated to give 15.7 g of product mixture containing the title compound.

Reference Example 8

N, N'-Di (benzyloxycarbonyl) -2 '- (0-acetyl) -6' - (n-octyl) -4 ', 5'-didehydro-spectinomycin

A dried 15 ml three-necked flask is fitted with a magnetic stir bar and a lid for serum storage vials. Tetrahydrofuran (10 mL) was added and dissolved in N, N'-di-benzyloxycarbonyl-2 '- (0-acetyl) -6'-methylene-4', 5'- (332 mg, 0.5 mmol). didehydro-spectinomycin and 25 mg of copper (II) bromide. The resulting slurry was cooled to -78 ° C under a nitrogen atmosphere and 1.2 ml of a 2.1 M solution of heptylmagnesium bronide was injected through said lid. TLC (1: 4 v / v methanol / chloroform) indicated that the starting material was transformed into a slightly less polar main product. The reaction mixture was then poured into a mixture of ethyl acetate (20 mL) and acetic acid (0.3 mL) and diluted with water (10 mL). After stirring, the organic phase was separated and washed with saturated aqueous sodium bicarbonate solution (15 mL) and brine (10 mL). The aqueous extracts were backwashed with ethyl acetate (20 mL), and the combined organic extracts were dried over sodium sulfate and evaporated to a foam. This was then purified by chromatography on 6.0 g silica gel, eluting with a 1: 9 by volume mixture of acetonitrile and chloroform. 0.32 g of pure title compound is obtained.

CMR (CD3COCD3): 183.1, 178.4, 170.0,

157.7, 138.0, 124.1,

128.3, 102.5, 96.0, 93.8,

75.6, 73.4, 67.3, 66.1,

59.7, 57.2, 35.0, 32.4,

-817

31.4, 30.0, 29.8, 29.6,

26.7, 23.2, 20.4 and 14.3 ppm.

Mass spectrum (after silylation) m / e: 856 (M '), 841, 769, 67],

600, 484, 356, 313, 284, 243, 217, 199, 171, 145 and 73.

Using the procedure described in Reference Example 8, but substituting heptyl magnesium bromide for an appropriately substituted alkyl magnesium bromide, the following compounds were obtained:

N, N'-di (benzyloxycarbonyl) -2'-0-acetyl-4 ', 5'-didehydro-6'-ethyl-apektÍnomicin

CMR: 183.1, 176.2, 170.0, 151.3, 137.81, 129.0,

128.1, 102.6, 95.9, 93.6, 75.3, 74.2,

67.3, 66.1, 60.0, 57.1, 36.8, 31.3, 20.8, 20.0 and 13.6 ppm.

N, N'-di (Benzyloxycarbonyl) -2'-O-acetyl-4 ', 5'-didehydro-6' - (n-propyl-spectinomycin CMR: 183.3, 176.5, 170, 1, 157.2, 138.2, 129.2,

128.4, 102.6, 96.1, 93.8, 75.6, 74.6,

67.4, 66.2, 60.1, 57.3, 34.8, 32.2, 28.8,

22.7, 20.9 and 13.9 ppm.

N, N'-Di (Benzyloxycarbonyl) -2'-O-acetyl-4 ', 5'-didehydro-6' - (n-butyl) -spectinomycin CMR: 183.1, 176.4, 170 , 0, 157.9 138.2, 129.1,

128.7, 102.5, 96.0, 93.8, 75.6, 74.6,

67.3, 66.1, 60.0, 57.2, 35.0, 31.7, 31.4,

26.3, 22.8, 20.9 and 14.1, ppm. N, N'-di (Benzyloxycarbonyl} -2'-O-acetyl-4 ', 5'-didehydro-6' - (n-pentyl) -epeptinomycin CMR: 183.1, 176.5, 170.0, 157.8, 138.4, 129.1,

128.3, 102.6, 96.0, 93.7, 75.6, 74.8,

67.3, 66.0, 60.2, 57.1, 35.0, 32.0, 31.4,

29.2, 26.6, 23.0, 20.9 and 14.2 ppm.

Reference Example 9

N, N'-di (benzyloxy-carbonyl) -6'-octyl spectinomycin

Dissolve 0.32 g in 12.0 ml of 2-propanol

N, N'-di (benzyloxycarbonyl) -2 '- (0-acetyl) -4', 5'-didehydro-6 '- (n-octyl) -spectinomycin, followed by 0.12 ml triethylamine is added. The reaction mixture was then divided into four equal portions and filled into test tubes containing 30 mg of platinum oxide each. The test tubes are then carefully placed in a Parr foil vessel and shaken under a hydrogen pressure of 1.4 at 2 for 2 hours. The catalyst was then filtered off and collected on the combined filtrate

O, 7 mL of water was added. After standing for 16 hours, the filtrate was heated for an additional three-quarters of an hour at 65 ° C and evaporated to give a residue which was chromatographed on 6 g of silica gel. Elution was carried out with 200 ml of 0.5% methanol in chloroform, 100 ml of 1% methanol in chloroform, 150 ml of 1.5% methanol in chloroform, 200 ml of 2% methanol in chloroform and 100 ml of 3% raetane in chloroform. The product-containing fractions were selected by analysis by thin-layer chromatography: the product gave a purple-red spot with a DNP spray followed by sulfuric acid spray and heating to give 0.09 g of the title compound.

CMR (CDaCOCDa): 201.5 , 157, 138.1, 129.1, 128.3 , 97.5 , 92.3, 75.1, 74.7, 74.6, 71.8, 67.2, 66.4, 65.6, 61.1, 60.1, 57.5, 44.1, 36.1, 32.4, 31.7, 31.4, 30.7, 30.1, 29.8, 25.7, 23.1 and 14.3, ppm. MS (silylation after) m / e: 928 (M *), 913, 901, 837, 823, 793, 745, 624 , 611,

539, 493, 359 and 91.

9a. PREPARATION

N, N'-Di (Benzyloxycarbonyl) -6'-propyl-spectinomycin in 0.7 g of 4 ', 5'-didehydro-2' - (0-acetyl) -N, N'-di is dissolved in tetrahydrofuran (benzyloxycarbonyl) -6'-propyl spectinomycin and the resulting solution was cooled to -78 ° C. To the solution was then added 3.0 mL of a 1 M solution of lithium tri (sec-butyl) borohydride in tetrahydrofuran. The resulting reaction mixture was stirred at -78 ° C under nitrogen for 25 minutes and then poured into a mixture of 10 mL of ethyl acetate and 0.2 mL of acetic acid. To the resulting mixture was added 10 ml of ethyl acetate, 10 ml of water and 1.0 ml of triethylamine. After 2 hours, additional triethylamine (1.0 mL) was added and the resulting solution was stirred at room temperature for 16 hours. 20 ml of Skellysolve B and 15 ml of saturated aqueous sodium bicarbonate solution are then separated and the phases are separated. The upper layer was washed with saturated aqueous sodium chloride (15 mL). The organic phase was dried over sodium sulfate and evaporated to an oil. The oil is then chromatographed on 40 ml of silica gel (silica gel is applied to the column with methylene chloride). The column was treated with 50 ml of methylene chloride, 1: 1 methylene chloride: chloroform (50 ml), 50 ml of chloroform, 100 ml of 0.5% methanol in chloroform, 100 ml of 2.5% methanol in chloroform and eluted with chloroform containing% methanol. 0.45 g (58.3%) of the title compound is isolated by evaporation of the appropriate fractions.

9 or 9a. as described in Reference Example 2, but starting from N, N'-di (benzyloxycerbonone) -2 '- (0-acetyl) -4', 5'-didehydro-6 '-octyl or propyl spectinomycin. The following compounds can be prepared using 6'-alkyl analogs:

-919

N, N'-di (benzyloxy-carbonyl) -6'-ethyl 8pektinomicin

CMR: 201.0, 158.2, 138.2, 129.1, 128.4, 97.6,

92.3, 75.2, 74.7, 74.6, 71.5, 67.2, 66.2,

65.6, 61.1, 60.2, 57.6, 57.3, 44.1, 38.2,

31.7, 31.4, 18.2 and 14.1 ppm. N, N'-di (benzyloxycarbonyl) -6 '- (n-propyl) -spektinomicin

CMR: 201.8, 156.7, 138.1, 129.1, 128.3, 97.5,

92.3, 75.1, 74.7, 74.6, 71.8, 67.2, 66.4,

65.6, 60.9, 59.9, 57.5, 44.1, 35.7, 31.4,

30.8, 27.7, 23.0 and 14.2 ppm. N, N'-di (benzyloxycarbonyl) -6 '- (n-butyl) -spektinomicin

CMR: 201.5, 156.2, 138.2, 129.4, 128.6, 97.7,

92.6, 75.3, 75.1, 74.8, 72.1, 67.6, 66.7,

66.0, 57.8, 44.3, 35.3, 31.6, 32.5, 32.1,

31.8, 31.2, 30.2, 29.2, 25.6, 23.3 and 14.6 ppm

N, N'-di (Benzyloxycarbonyl) -6 '- (n-pentyl) -spectinoracin

CMR: 202.0, 158.1, 138.2, 129.1, 128.4, 97.6,

92.4, 75.2, 74.6, 74.0, 71.8, 67.2, 66.4,

65.7, 61.1, 60.1, 57.5, 57.3, 44.1, 36.1,

32.3, 31.7, 31.4, 29.7, 25.6, 23.2 and 14.3 ppm.

N, N'-di (benzyloxycarbonyl) -6 '- (cyclohexylmethyl -benzoic spectinomycin

CMR: 202.0, 157.5, 138.0, 129.1, 178.3, 97.5,

92.3, 75.1, 74.7, 72.1, 67.2, 66.4, 65.6,

60.9, 60.7, 57.5, 57.3, 44.1, 38.2, 33.8,

33.5, 31.7, 31.4, 27.2 and 26.9 ppm.

Example 1

6 '- (n-Octyl) -spektinoraicin

Dissolve 0.09 g of N, N'-di (benzyloxycarbonyl) -6 '- (n-octyl) -spectinomycin in 7.0 ml of methanol and stir the solution in a nitrogen atmosphere. Then 60 ml of palladium on carbon and 0.45 ml of 97% formic acid were added. After vigorous stirring for 40 minutes, the catalyst is filtered off and the filtrate is concentrated under high vacuum. The residue is taken up in 8 ml of water and the pH of the aqueous solution is adjusted to 1.5 by the addition of 0.35 ml of 0.1 N hydrochloric acid. The aqueous solution was then freeze-dried to obtain a sample for CMR uptake. After uptake of the CMR, the sample was diluted with water and freeze-dried to give the title compound as its hydrochloride.

CMR (DjO, cyanomethane internal reference):

92.9, 92.6, 91.1, 70.9, 68.8, 65.1, 64.7,

60.6, 58.7, 57.5, 38.2, 33.4, 30.8, 30.1,

29.5, 28.5, 28.2, 24.2, 21.5, 12.8 ppm.

By following the procedure described in Example 1, but employing, instead of N, N'-di-benzyloxycarbonyl-6 '- (n-octyl) -epectinomycin, the appropriately substituted protected 6'-alkyl spectinomycins, the following compounds were obtained: in the form of hydrochloride salts:

6'-ethyl-8pektinomicin

CMR: 29.8, 29.4, 60.6, 58.6, 57.6, 64.6, 70.7,

65.0, 92.7, 92.7, 91.0, 58.5, 68.8, 35.0, 16.6 and T.2.4 ppm.

G'n- (n-propyl) -spektinomicin

CMR: 29.8, 29.4, 60.6, 58.6, 57.6, 64.6, 70.9,

64.9, 92.6, 92.6, 91.0, 38.5, 68.7, 32.5,

25.4, 21.0 and 12.3 ppm.

6 '- (n-butyl) -spektinomicin

CMR: 29.9, 24.4, 60.4, 58.5, 57.4, 64.5, 70.7,

64.9, 92.5, 92.5, 90.9, 38.7, 68.7, 32.9,

30.1, 23.1, 20.4 and 12.5 ppm.

6 '- (n-pentyl) -epektinomicin

CMR: 29.8, 29.3, 60.6, 58.6, 57.7, 64.7, 71.0,

65.0, 92.7, 92.7, 91.1, 38.6, 68.8, 32.9,

30.1, 23.2, 21.0, 20.8 and 12.5 ppm.

'- (cyclohexylmethyl) -spectinomycin

CMR: 30.1, 29.5, 60.6, 58.6, 57.5, 64.7, 70.9,

65.1, 92.6, 92.6, 91.1, 38.2, 68.8, 33.4,

30.7, 28.5, 28.5, 28.2, 28.2, 24.2, 21.5 and 12.8 ppm.

6 '- (n-undecyl) -spektinomicin

CMR: 60.5, 58.6, 57.5, 64.9, 70.9, 65.2, 97.7,

92.8, 91.2, 38.8, 68.8, 33.4, 30.9, 28.4,

28.8, 28.8, 28.8, 28.8, 28.8, 28.8, 24.4,

21.5, 12.8, 30.2 and 24.6 ppm.

The resulting compounds are compounds of formula (Ia) on the basis of the CMR spectrum.

Example 2

6 '- (n-propyl) sulfate pentahydrate -spektinomicin

2.2 g of N, N'-di-benzyloxycarboryl-6 '- (propyl) -spectinomycin are dissolved in 100 ml of methanol and the resulting solution is stirred under nitrogen. Then 1.39 g of palladium on carbon was added to the solution followed by 4.3 ml of 97% formic acid. added. After vigorous stirring, the catalyst is filtered off and the filtrate is evaporated under high vacuum. The residue is taken up in 13 ml of water and the resulting solution is acidified with 3.5 ml of 1M aqueous sulfuric acid. The sample was then freeze dried and recrystallized from acetone / water to give the title compound (0.87 g).

CMR (D10, cyanomethane internal standard): 118.07, 92.67, 91.24, 91.06, 70.73, 68.94, 65.01, 64.74, 60.78, 59.15 , 57.69, 38.77, 32.83, 30.35, 29.77, 25.78, 21.18 and

12.45 ppm.

Claims (6)

PATENT CLAIMS A process for the preparation of spectinomycin analogs of formula (I): wherein: R 'is C 1 -C 12 alkyl or C 4 -C 8 cycloalkyl-C 1 -C 4 alkyl, and -1021 for the preparation of their pharmaceutically acceptable acid addition salts and hydrates of the latter, characterized in that a compound of formula XV wherein R'j and R 'are a protecting group is aralkoxycarbonyl, halogenated alkoxycarbonyl or alkoxycarbonyl. and R10 is an acyl group with a compound of formula R 'Mg X wherein R' is as defined in formula I and X is halogen to form a compound of formula III wherein R 'is in the formula (I), while the other substituents have the same meaning as in the formula (IV), the compound of formula (II) in which the substituents have the meaning given in the formula (III): The protecting groups of 20 and R '' are cleaved and, if desired, the resulting general formula (I) The compound of formula I is converted into a pharmaceutically acceptable acid addition salt thereof or the bridging salt of the latter. Process for the preparation of compounds of formula (Ia) according to claim 1, wherein R 'is as defined in claim 1, characterized in that the appropriate starting materials are used. ^ 0 The process according to claim 2, 6'-ethyl-spectinomycin, 6'-ethyl-spectinomycin-dihydrochloride, 6 '- (n-propyl) -spectinomycin, 6 '- (n-propyl) -spectinomycin dihydrochloride,?' - (n-pentyl) -spectinomycin, 6 '- (n-pentyl) -spectinomycin-dihydrochloride, 6' - {n-octyl) -spectinomycin, 6 ' - (n-octyl) -spectinomycin dihydrochloride, 6 '- (n-undecyl) -spectinomycin, 6' - (cyclohexylmethyl) -bpectinomycin and 6 '- (cyclohexylmethyl) -spectinomycin dihydrochloride, characterized in that: using the appropriate starting materials. The process of claim 2, 6 '- (n-propyl) -spectinomycin sulfate, t'-isobutyl-Bpectinomycin, 6'-isobutyl-spectinomycin dihydrochloride 6 '- (3,3-dimethylbutyl) -Bpektinomicin, 6 '- (3,3-dimethyl-butyl]) - 8pektinomicin dihydrochloride 6 '- (cyclopentylmethyl) -spectinomycin and?' - (cyclopentylmethyl) -spectinomycin dihydrochloride, characterized in that the appropriate starting materials are used. 5. A process for the preparation of a pharmaceutical composition comprising the steps of: a compound of formula (I) obtained by the process of claim 1 wherein: R 'is C 1 -C 12 alkyl or C 3 -C 8 cycloalkyl (C 1 -C 4) alkyl, or a pharmaceutically acceptable acid addition salt or hydrate thereof, in admixture with pharmaceutically acceptable carriers and / or excipients. 6 sheets drawing The Director of Economic and Legal Publishing is responsible for the publication 89,921.66-4 Alföldi Nyomda Debrecen - Chief Executive Officer: István Benkő Chief Executive Officer