NL8001315A - SPECTINOMYCIN AND SPECTINOMYCIN ANALOGA, METHODS FOR PREPARING THE SAME AND PHARMACEUTICAL PREPARATIONS PREPARED THEREOF. - Google Patents

Description

*

Spectinomycin and spectinomycin analogs, methods of preparing the same and pharmaceutical preparations prepared therefrom *

The invention relates to novel anomers and asteric mixtures of spectinomycin analogues, methods of preparing the same and pharmaceutical preparations prepared therefrom, as well as to intermediates suitable for use in the aforementioned methods.

Spectinomycin is an antibiotic of the formula 59. So far, spectinomycin has been prepared by a microbiological method. See U.S. Patent 3,234,092 *

Some analogues of spectinomycin have been described by Rosenbrook Jr et al in J. Antibiotics, 28, p. 963 and 960 (1978) and J. Antibiotics, 31_, p. 451 (1978). In addition, Camey et al., Describe chlorodoxides of spectinomycin in J. Antibiotics, 30, 960 (1977). Furthermore, 9-epi-4 (R) -dihydrospecti-nomycin is described by Foley et al. In J. Org. Chem., 43, 22, pp. 4355-4359 (1978).

However, no biological activity is described for the spectinomycin analogs and their derivatives mentioned in the above references.

The reaction on the 5-hydroxyl of 2-deoxvstrepta-20 τΗπο (i) with tri-0-acetyl-2-deoxy-2-nitroso-aD-glycopyranosyl chloride (2) to an α-pseudodisaccharide (3) has been described by Lemieux in Can. J. Chem. 51, p. 53 (1973) and shown in Scheme A, wherein CBz is a carbobenzyloxy group. The extension of this reaction for the preparation of di- and tri-saccharides has been described by Mallams et al in J. Chem. Soc. Perkin I, p. 1118 (1976),

Oxime removal has been described by Lemieux et al, Can. J. Chem., 51, 19 (1973) and Mallams et al., J. Chem. Soc. Perkin I, p. 1097 (1976).

The anomers and asteric mixtures of the new 800 1 3 15 2 spectinomycin analogues of the invention are of Formula 1, wherein R represents a group of Formula 8 or Formula 9 wherein R 1 through R 4 are the same or different and hydrogen , represent a lower alkyl, lower alkenyl, lower haloalkyl, lower aminoalkyl, lower alkynyl, OX or - {C4 ^ - OX, with the proviso that R 1 and R 2 are not a hydroxyl group and one of the groups R and R 1 must be hydrogen, wherein X is hydrogen, a lower alkyl, lower alkenyl, or lower alkynyl group? η is 1, 2, 3 or 4?

Rj. is hydrogen or a lower alkyl group?

Rg to R ^ 4 represent hydrogen, a lower alkyl, lower alkenyl or lower alkynyl group, with the proviso that one of the groups R ^ and Rg and one of the groups R ^ j and R ^ is always hydrogen , B and Bj are the same or different and are hydrogen, a hydroxyl, alkoxy, o-lower alkenyl, thio, thio-lower alkyl or thio-lower alkenyl group and A is oxygen or sulfur, with the proviso that when the compound is of formula 10 Bj cannot be a hydroxyl group, the numbering of the carbon atoms indicated in formula 1 is used hereinafter.

The compounds according to the invention comprise the hydrate forms of compounds of the formula 1. These compounds are hydrated in the 3 'position and have the formula 1', wherein A, B, Bj, R and Rg to RJ4 are as indicated above. have meaning. The invention further includes pharmaceutically acceptable salts of the compounds of formulas 1 and 1 '.

The group - (CH-) includes straight-chain lower alkyl groups and isomers thereof.

Low alkyl includes the methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl group and the isomeric forms thereof.

Low alkenyl includes ethylidene, propylidene, 800 1 3 15 * V * 3 butylidene, pentylidene, hexylidene, heptylidene, octylidene and the isomeric forms thereof.

Lower alkynyl includes ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl and the isomeric forms thereof.

Low alkoxy includes methoxy, ethoxy, propoxy, butoxy, pentoxy, hexoxy, heptoxy, octoxy and the isomeric forms thereof.

Acyl includes formyl, acetyl, propionyl, butyryl, and pentionyl and isomeric forms thereof.

Aralkyl includes benzyl, phenethyl, fenpropyl, fenbu-tyl, fenpentyl, diphenylmethyl, diphenyloctyl and isomeric forms thereof.

Low haloalkyl includes - (CH_) -halogen and isomeric forms thereof.

The group contains 1-3 halogen substituents.

Low aminoalkyl includes a group of the formula 11 and isomeric forms thereof.

Aroyl includes benzoyl, substituted benzoyl, naphthoyl, and substituted naphthoyl. The substituted benzoyl and naphthoyl may contain 1-3 substituents such as lower alkyl, lower alkoxy, nitro or halogen.

Halogenated alkoxycarbonyl includes mono-, di-, tri-halomethoxycarbonyl; mono-, di-, tri-haloethoxycarbonyl; mono-, di-, tri-halopropoxycarbonyl; mono-, di-, tri-halo-butoxycarbonyl; mono-, di-, tri-halo-pentoxycarbonyl and isomeric forms thereof.

Halogen includes the fluorine, chlorine, bromine and iodine atom.

Aralkoxycarbonyl includes benzyloxycarbonyl, phenethoxycarbonyl, fenpropoxycarbonyl, fenbutoxycarbonyl, fenpentoxy-carbonyl, diphenylmethoxycarbonyl, diphenyloctoxycarbonyl and isomeric forms thereof, and fluorenylmethoxycarbonyl.

Alkoxycarbonyl includes isopropyloxycarbonyl, tertiary butyloxycarbonyl and tertiary pentyloxycarbonyl.

When more than one hydroxyl or alkoxy group are present on the sugar residue, they can be the same or different.

800 1 3 15 4

Of significance is the stereochemistry at the glycosidic bond, which is the 1 'position in the formula 1.

"α-Anomer" means a 1 'substituent behind the plane of the ring system and "β-anomer" means a 1' substituent behind the plane of the ring system. In particular, "β-Anomer" means anomers with the C-1 'configuration corresponding to spectinomycin.

Compounds of the invention having the desired biological activity are β-anomers of the compound 1. This glycosidic configuration is shown in formula 59. Thus, to obtain biologically active analogues of spectinomycin, selectivity at the 1 'position is desirable.

Actinamines and actinamine derivatives include the aminocyclitols of the formula 6.

Strikers include pyran, natural and synthetic sugars, chirals and achirals.

The desired selectivity is achieved with the method according to the invention. Heretofore, it has not been possible to obtain a wide variety of spectinomycin analogues by preparing a 6-membered hemiketal having a spectinomycin structure with biological activity by converting δ-hydroxy oximes to δ-hydroxy ketones.

With regard to the process according to the invention, it has been found that a starting actinamine with protected amino groups reacts at the C 1 -OH site without protecting the other α or 8-OH groups at the C 2, c 4 or C 5 sites. need be omitted, with which a difficult step to perform can be omitted, which in itself is surprising. Another advantage of the method according to the invention is the selective formation of a C3'-carbonyl group or a group from which a C3'-carbonyl group can be formed by an elimination reaction. The C31 carbonyl group is of great significance but difficult to realize in spectinomycin analogues.

The reaction according to the invention is shown in scheme B, in which 800 1 3 15

V

5 R represents a group of the formula 8 or 9,

Rj to r4 may be the same or different and represent hydrogen, a lower alkyl, lower alkenyl, lower alkynyl, lower haloalkyl, lower aminoalkyl, -Οχ or - (CH.) -OX, with

X »XI

It should be understood that R 1 and R 1 do not represent a hydroxyl group and one of the groups R 1 and R 1 must be hydrogen; X is hydrogen, a lower alkyl # lower alkenyl or lower alkynyl group; n is 1, 2, 3 or 4; 10 Rj. is hydrogen or lower alkyl group;

Rg to R4 represent hydrogen, a lower alkyl, lower alkenyl or lower alkynyl group; R * 7 'R'a' R011 6.1 R'l2 6611 low a3Jcy1 '·' low alkenyl, low alkynyl, a protective aralkoxycarbonyl, halogenated alkoxycarbonyl or alkoxycarbonyl group, provided that one of the groups and Rg and one of the groups Rjj and Rj2 is always hydrogen and further with the proviso that one of the groups R'j and R'g and one of the groups R'jj and R'12 is always a protecting group is; R 1 is an alkyl, aralkyl or aroyl group; 16 A is oxygen or sulfur; and B and Bj are the same or different and represent hydrogen, a hydroxyl, alkoxy, o-lower alkenyl, thio, thio-lower alkyl or thio-lower alkenyl group.

An example of a reaction according to the invention is shown in scheme C.

* »

Although both α and β anomers can be formed in step 1 of the above-mentioned process, the β anomers can preferably be obtained by separating the 8- from the α anomers after any step of the process. Also, the use of an available enantiomeric sugar in the initial coupling reaction to obtain a large amount of β-anomer, or epimerization of an intermediate from end product, can increase the amount of β-anomer obtained. Furthermore, asteric mixtures 35 themselves can be used as bactericides, since the biological activity is 800 1 3 15 6 due to an action anomer present therein.

Anomers and asteric mixtures of a compound include spectinomycin analogues within the scope of the invention with bactericidal activity. Although the 8-configuration is the active anomer according to the invention, the term "anomers and asteric mixtures" is not limitative, since new a-anomers may also be present, without prejudice to the activity of an asteric mixture. Also, α-anomers of the spectino-mycine analog can in some cases advantageously be anomerized to the active form of the analog. Therefore, α configuration is not excluded at any step of the method of the invention.

On the other hand, compounds useful in the invention are products of the formula 1 having the $ configuration, since these anomers exhibit bactericidal properties. Separation of the anomers can be performed after each step of the process. Preferred β-Anomers are compounds, including C-2 and C-6 hydroxyl groups, of the formula 12, wherein all substituents have the meanings indicated above.

The invention further relates to new intermediates for the method according to the invention. These include: (i) anomers and asteric mixtures of compounds of the formula 2, wherein R represents a group of the formula 8 or 9,

Rj to R4 may or may not be the same and represent hydrogen, a lower alkyl, lower haloalkyl, lower aminoalkyl, lower alkenyl, lower alkynyl, -OX or - (CH_) -0X, provided that R 1 and R 1 are not OH and one of the groups R 1 and R 4 must be hydrogen; X is hydrogen, a lower alkyl, lower alkenyl or lower alkynyl group; η is 1, 2, 3 or 4; R ,. is hydrogen or lower alkyl group; R6, Rg, Rjq, R13 and R14 represent hydrogen, a lower alkyl, 800 1 3 15% 7 lower alkenyl or lower alkynyl group; R (is hydrogen or lower alkyl group; R'7, R'g, R'jj and R'12 is a lower alkyl, lower alkenyl, lower alkynyl, a protective aralkoxycarbonyl, halogenated alkoxycarbonyl or alkoxycarbonyl group propose that one of the groups R'7 and R'g and one of the groups R'jj and R * 12 is always a blocking group;

R15 is hydrogen or acyl group;

Rjg is a lower alkyl, aralkyl or aroyl group; 10 A is oxygen or sulfur; and B βη represent hydrogen, hydroxyl, alkoxy, o-lower alkenyl, thio, thio-lower alkyl or thio-lower alkenyl group, with the proviso that when the compound has the formula 13 wherein CB 2 is a carbobenzyloxy group , Bj cannot represent hydrogen or hydroxyl group; (ii) anomers and asteric mixtures of compounds of the formula 3b, wherein A, B, B ^, R ^ to Rg, Rg, R'7, R'g, Rg, Rjg, R '^, R'12 R13 and R14 have the meanings indicated above and Rjg is a lower alkyl, aralkyl or aroyl group, with the proviso that R1, R2 and Rg do not represent a hydroxyl group.

In compounds of formula (3b), the levl-ketal may be wholly or partly in the open ketone form * These two forms may be in equilibrium, as shown in scheme D.

Spectinomycin is an aminocyclitol antibiotic with a unique structure in that it is a single sugar component bound to an actinamine by both a β-glycosidic and a hemi-metal bond. The method according to the invention for the preparation of the analogs with this unique bond is a synthesis, in which a sugar derivative and a protected actinamine are coupled. The sugar can be natural, synthetic, chiral or achiral sugar.

The method consists of two basic steps, as shown in the above diagram. Step 1 consists of the sub-steps according to the scheme E.

800 1 3 15 8

Other examples of step 1 are shown in scheme P.

Before step 1a, the two amino groups of the actinamine derivative 6a are protected by each protecting with a protecting group such as an aralkoxycarbonyl, haloalkyloxycarbonyl, aryloxycarbonyl or alkoxycarbonyl group, which are known above. For preparation and deprotection of carbo-benzyloxy and carbo-tert-butyloxy derivatives of amino acids, see RA, Boissinas Chapter, Selectively Removable Amino 10 Protective Group used in the Synthesis of Peptides, in: Advances in Organic Chemistry, 3 : 159-190 (1963); for the use of the tert.-butyloxycarbonyl group to protect amine to ALDRICH Technical Information Bulletin entitled BOC-OC (September 1976); and using the trichloroethoxycarbonyl group to protect amines after Windholz et al., Tetrahedron Letters, 2555 (1967). The actinamines are prepared by known methods, see, for example, Suamin et al., Bull. Chem. Soc. Jap. 43, 1843 (1970).

The sugars are commercially available or can be prepared by known methods, for example, the method described by Mochalin et al., Chem. It. Comp. 699 (1977) (English translation of KHIM Geterotsiklsoedin, 867 (1977)).

Step 1a involves a coupling reaction between actin "nriru" 6 and sugar 7a, 7b or 7c. This step occurs in a solution of Ν, Ν-dimethylformamide or a similar solvent, such as diethyl ether, tetrahydrofuran or dimethoxymethane, sometimes in the presence of a base. The reaction is best performed under a nitrogen atmosphere at room temperatures and pressures, as described for a similar reaction by Lemieux et al., Can. J. Chem., 51, 53 (1973). The temperature range of the reaction is generally between 0 and 45 ° C with molar ratios of activated sugar in solvent between 0.01 M and 0.5 M added to actinamine in a solution at a concentration of 0.01 M to 0 .5 M so that in the reaction mixture the molar ratio of sugar to actinamine is between 0.2 and 4. Preferred reaction conditions are a temperature of 20-30 ° C, using 800 1 3 15 9 dimethylformamide as a solvent with the ratio of sugar to actinamine between 3: 2 and 2: 3. The reaction time can be between 4 hours and 1 week, but is preferably between 24 and 48 hours.

The oxime 5a, 5b or 5c obtained is generally separated from the reaction mixture by concentration or by concentration and vigorous stirring with excess water. The solid obtained is taken up in chloroform and evaporated to dryness to yield the crude oxime intermediate, α and β-Anomers can be further fractionated by chromatography on a silica gel column, eluting with methanol in chloroform in the ratio of 1:99 to 2:98. However, the use of normal recovery methods such as extraction, crystallization, chromatography and combinations thereof are encompassed by the method of the invention. In step 1b, the oxime group is removed from compound 5, whereby a cyclic hemital, the above compound 4 is obtained. The reaction of step 1b is performed by deoximation methods similar to those described by Lemieux et al. In Can. J. Chem., 51, 19 (1973) and by Mallams et al. In 20 J. Chem. Soc. Perkins I, 1097 (1976). The deoximation reaction is carried out, for example, by dissolving either the crude oxime or the α and β oximes of compound 5 to be separated in a solvent, followed by addition of acetaldehyde and hydrochloric acid. The initial concentration of the oxime in solvent is between 0.01 M and 1.5 M, but preferably between 0.1 M and 0.3 M and the mole. - the ratio of the aldehvd to the oxime is between 1: 1 and 80: 1.

IN hydrochloric acid is added in a 2: 3 ratio with the oxime. The reaction mixture is stirred at room temperature for 3.75 hours to 5 days, after which sodium bicarbonate can be added and stirring is continued for an additional 15 minutes. The concentrate can also be directly chromatographed, so that silica gel removes the HCl during purification.

Intermediate 4 is recovered in conventional ways as described above. One way is by preparing a filtrate which is evaporated to dryness in vacuo to yield the crude 800 1 3 15 10 product. This product can be fractionated again several times by silica gel chromatography using chloroform and methanol as eluents in the ratio of about 95: 5. Combination of similar fractions corresponding to the thin layer chromatographic analysis thereof, followed by evaporation to dryness in vacuo, gives separate hemiketal et and β anomers, which are included in Structural Formula 4.

Bases other than sodium carbonate can be used in the deoximation of step 1b. However, these should be chosen so that acetates are not attacked, ie hydroxides cannot be used in excess. Suitable bases are benzoates or dipotassium hydrogen phosphate.

Solvents that can be used include acetonitrile, tetrahydrofuran, ether or dimethylformamide. The preferred solvent is acetonitrile.

In step 1c, both 1 or 2 substituents on the sugar residue of the compound 4 are removed, and a C-3 'carbonyl is formed by 6-elimination of the C-4' site, whereby the compound of the formula 3 is obtained. A second elimination can occur on c-6 ', whereby a C-4', c-5 'olefin is obtained. This step is performed in the presence of a base system at temperatures between 0 and 80 ° C for about 2 hours to 1 week. Base systems which can be used include potassium bicarbonate, triethylamine, pyridine and alkoxide. A preferred system is potassium bicarbonate / acetonitrile. 0-20 Mol.eq. base can be used, but 1-10 is preferred.

The manner in which step 1c is carried out also depends on the particular protecting groups on the sugar residue and on the actinamine residue of the intermediate 4. In general, the protecting groups on the sugar residue are easier to remove than those on the actinamine residue. Step 1c is a novel method for the mild and selective formation of the major c-3 'carbonyl group as such or in latent form by elimination.

For example, acetates eliminate acetic acid, benzoate, benzoic acid, benzyl ethers eliminate benzolate, halides 800 1 3 15 11 eliminate hydrogen halide. These are not limiting examples of the elimination occurring in step 1c.

The intermediates according to the invention, in particular the products from step 1c, are useful substances for the synthesis of a variety of analogues. This is done by changing the functional groups of the intermediates by known methods such as halogenation, reduction, oxidation, chain elongation, etc.

In some instances in the present invention, the elimination step 1c involves migration of the O 3 'substituent on oxygen to the C-2' oxygen to form a c-3 'carbonyl group. This behavior is explained in the conversion from 4a to 3a and from 4c to 3c. In other instances, the C-3 * substituent on oxygen does not migrate to form a masked or latent C-3 * carbon-15-yl; these are enol derivatives. An example is the conversion from 4b to 3b. The enol derivatives can be hemiketals or open ketone isomers or mixtures of these forms.

Both of the above cases are new, useful # selective methods, yielding final spectinomycin analogs with 20 C-3 * carbonyl groups. The masked or latent C-3 'carbonyl groups containing intermediates have unique chemical properties which make them useful for modification by known methods such as halogenation, alkylation, acylation, oxidation and the like. Finally, the masked or latent c-3 'carbonyl group is much more stable, especially with respect to base, so that it is part of the more versatile and easier to separate intermediates.

The compound of formula III is removed from the reaction mixture by conventional methods such as precipitation, crystallization or concentration followed by chromatography.

In step 1d, the protection is removed at one or several places of the sugar ring. Usually these are C-2 ', c-3' or c-6 'and depending on the nature of the protecting groups, acid and / or base can be used. When base is used, such as in the conversion of 3a to 2a or from 3c to 2c, 800 1 3 15 12 hydrolysis is carried out at -10 to + 50 ° C for 5 minutes to 40 hours. Preferred conditions , are 20-30 ° C for 1-20 hours.

Alcohols that can be used include methanol, ethanol and isopropanol, but methanol is preferred. Any base that does not affect the quality of the product can be used. These include sodium bicarbonate, potassium bicarbonate, pyridine, dipotassium hydrogen phosphate, triethylamine, sodium potassium tartrate, but the preferred catalyst is dipotassium hydrogen phosphate. In the first step of the two-step process for converting 3b 'to 2b, the C-61 acetyl group is selectively removed using the above-mentioned basic alcoholysis conditions, while the c-3' methoxyl group remains intact.

Acid catalysis can also be used to remove the protecting groups from the sugar ring. For example, after the C-6 'acetyl group is removed from 3b' using base as described above, the residual protection from C-3 'can be removed by acid treatment to yield 2b. Also, 3b 'to 2b can be converted in one step using acid catalysis.

Deprotection using acid is usually performed at 0-80 ° C, preferably 20-30 ° C for 1 hour to 3 days, preferably 2 hours to 2 days. Acids such as hydrochloric acid, paratoluene sulfuric acid or phosphoric acid can be used here; hydrochloric acid is preferably used. Solvents can include tetrahydrofuran, aqueous dimethoxyethane, methanol or ethanol. Methanol or aqueous tetrahydrofuran is preferably used.

In some cases it is advantageous to combine step 1d with step 2 by using a reaction medium in step 2, which satisfies the deprotection conditions indicated in step 1d described above. For example, when step 2 is carried out in isopropanol with pyridine added, the intermediate 3a is converted to the antibiotic 800 1 3 15 13 spectinomycin. This shows that in addition to deprotecting the nitrogen (and C-4 ', C-5' saturation) that occurs under the influence of palladium catalyst and hydrogen, the C-2 'acetyl group is removed by alcoholysis due to the basic Solvent-5 medium system.

In some instances, it is advantageous to omit step ld or part of ld so that deprotection occurs in the biological system, releasing the active analog.

The particular conditions for deprotecting the actinamine residue in step 2 depend on particular groups, ie, groups R '^ or R'g and R' 'or R' 'ie protecting the amine on the actinamine. Also, by a suitable choice of R'j, R'g, R '^ and R' ^ and suitable choice of known deprotecting conditions, a C-4 ', C-5' olefin may remain intact or during removal of protection is reduced. When this group is benzyloxycarbonyl or aralkoxycarbonyl, deprotection can be carried out under a pressure of 0.07 to 14 at nitrogen over a conventional catalyst such as palladium carbon black, palladium on carbon, palladium on barium sulfate or pallaium on barium carbonate, while it is suspended in a solvent, for example, isopropanol, absolute ethanol, ethyl acetate, toluene or tetrahydrofuran.

Also can removing protections from connections where R '? whether R'g and R '^ or R'12 are an alkoxycarbonyl or aryloxycarbonyl group are carried out in the presence of acid in solution, such as nitromethane and methylene chloride.

When R '? whether R'g and R'jj or R'j represent a halo-alkoxycarbonyl group, deprotection is preferably carried out in the presence of zinc.

Any known method can be used to separate an analog or asteric mixture of a compound of formula 1 and the methods described here are not limiting. When the separation is performed under anhydrous conditions, compounds having a carbonyl group in the 3'-position 800 1 3 15 14 (formula 1) are obtained. When carried out under aqueous conditions, hydrated compounds are obtained at the 3 'position (formula 1'). In one such method, the excess solvent is evaporated and a crystalline salt of the compound is formed. These salts can be formed using a solution of an acid such as toluene sulfonic acid, hydrochloric acid, hydrobromic acid, hydroiodic acid or other acids in a solvent such as water, methanol, ethanol, isopropanol, ether, 1,2-dimethoxyethane or p -dioxane. The salt is separated by filtration and direct crystallization or by evaporation of the solvent, followed by recrystallization from a suitable solvent.

The crude analogs can also be purified by adsorption on a column of a weakly acidic ion exchange resin, such as Amberlite IRC-50 or CG-50, followed by elution with a solvent, such as water, methanol, ethanol, ether, tetrahydrofuran 1,2-dimethoxy, 1,2-dimethylethane, or p-dioxane, which contains hydrochloric, hydrobromic, hydroiodic and sulfuric acids.

Similarly, the salts of hydroxy analogs can be converted back into the free analog by dissolving the salt in solvent, such as water, methanol, ethanol, tetrahydrofuran, or 1,2-dimethoxyethane through a basic ion exchange resin, such as Dowex 1-XB (OH-) and evaporate the eluate containing the analogs with free base substituents.

Any step of the above-mentioned method can be applied to asteric mixtures of different anomers or to the desired β-anomer itself, which has been obtained by resolution or separation at any stage of the method. The remaining steps can be performed on β intermediates and lead to the desired biologically active anomers.

The preferred method is to separate β anomers from the mixture obtained in step 1, couple sugar and actinamine and carry out step 2 of the process on the β anomers, using only spectinomy-35 cine analogues which are biologically active.

800 1 3 15 15

Separation of anomers from asteric mixtures can be carried out by conventional resolution methods.

For example, a compound 4 can be separated to obtain a desired 3-component by chromatography on a silica gel column, eluting with a mixture of methanol in chloroform in the ratio of 1:99 to 2:98. Likewise, the separation of B anomers can be performed on an asteric mixture of compound 5 by combining 8 fractions obtained from a silica gel chromatography using a mixture of chloroform 10 and methanol as the eluent. Subsequently, evaporation to dryness in vacuo gives a separated hemi metal with the 3-structure.

Another particularly effective method according to the invention, and therefore preferred, is to obtain a relatively high concentration of a 3-anomer in the coupling reaction in step 1 by using an enomeric sugar, which forms the 3 structure. For example, D-arabinosa gives α and 3 oximes in a ratio of about 4: 1. In addition to avoiding costly and time-consuming separation methods on intermediates or products obtained in obtaining the 3 configuration, the enantiomer can also be available in inexpensive amounts. The use of such an enantiomer thus results in a mixture which is considerably enriched with the desired 3-anomer, which can be purified by a method as described above or used without further purification.

Acid salts can be prepared by neutralizing compounds of the formula 1 with the appropriate acid to below pH about 7.0 and advantageously to about 2-6 pH. Suitable acids include hydrochloric, sulfuric, phosphoric, sulfamic, hydrobromic, and the like. Acid and base salts of the compounds can be used for the same biological purposes as the parent compound.

Compounds of the formula 1 prevent the growth of microorganisms in different environments. For example, the compounds of formula 1 with the 3 configuration are active against Escherichia coli and can be used to reduce and stop sludge formation and sludge removal in paper mill systems, due to their bactericidal activity against this microorganism. * These β-anomers can also be used to extend the lifespan of cultures of Trichomonas fetus, Trichomonas hominis and Trichomonas vaginalis by removing contamination from Escherichia coli. Furthermore, β anomers are active against Bacillus subtllis, so that they can be used to minimize or prevent odor in fish or fish crates caused by this organism. The 10 anomers can also be used for the collection of laboratory tables and equipment in a mycological laboratory. 3-Anomers are also effective against Klebsiella pneumoniae.

The compounds of formula 1 are also suitable for treating bacterial infections, such as gonorrhea and tumors in humans and animals.

The compositions of the invention are administered to humans and animals in unit dosage forms such as tablets, capsules, pills, powders, granules, sterile parenteral solutions or suspensions, eye drops, oral solutions or suspensions and water-in-oil emulsions containing appropriate amounts of the compound of the formula 1.

For oral administration, either solid or flowable unit dosage forms can be used. To prepare solid preparations, such as tablets, the compound of formula 1 is mixed with conventional ingredients such as talc, magnesium stearate, dicalcium phosphate, magnesium aluminum silicate, calcium sulfate, starch, lactose, acacia gum, methyl cellulose and functionally similar substances as pharmaceutical diluents or carriers. Capsules are prepared by mixing the compound with an inert pharmaceutical diluent and placing the mixture in an appropriately sized hard gelatin capsule. Soft gelatin capsules are prepared by mechanically encapsulating a suspension of the compound with an acceptable vegetable oil, slightly liquid petrolatum or other inert oil.

Liquid unit dosage forms for oral 800 1 3 15 17 administration such as syrups, elixirs and suspensions can be prepared. The water-soluble forms can be dissolved in an aqueous carrier together with sugar, aromatic flavors and preservatives to form a syrup. An elixir 5 is prepared using a hydroalcoholic (ethanol) carrier with suitable sweeteners, such as sugar and saccharin, together with an aromatic flavoring agent.

Suspensions can be prepared with an aqueous carrier using a suspending agent such as gum acacia, tragacanth, methyl cellulose and the like.

For parenteral administration, liquid unit dosage forms are prepared using the compound and a sterile carrier, water being preferred.

The compound, depending on the carrier and concentration used, can either be suspended or dissolved in the carrier. In preparing solutions, the compound can be dissolved in water for injection and sterilized on the filter before being placed in a suitable vial or ampoule and sealed.

Advantageously, aids such as a local anesthetic, preservative and buffering agents can be dissolved in the carrier. to increase stability, the composition can be frozen after it has been placed in the vial and the water removed under vacuum. The dry lyophilized powder is then sealed in the vial and a vial of water for injection is provided to reconstitute the liquid 25 times per use. Parenteral suspensions can be prepared in much the same way, but the compound is suspended rather than dissolved in the carrier and sterilization cannot be performed by filtration. The compound can be sterilized by exposing it to ethylene oxide before suspension in the sterile vehicle. Advantageously, a surfactant or wetting agent is included in the composition to facilitate even distribution of the compound.

A rectal suppository can also be used to deliver the active compound. This dosage form is of particular interest when the mammal cannot be treated well with other dosage forms, such as orally or by insufflation, such as in young children or debilitated persons. The active compound can be incorporated in one of the known suppository bases in a known manner. Examples of such bases include cocoa butter, polyethylene glycols (Carbo waxes), polyethylene sorbitan, monostearate, and mixtures thereof with other compatible substances to change the melting point at the dissolution rate. These rectal suppositories can weigh about 1-2.5 g.

A heroine dosage form is here understood to mean physically discrete units suitable as unit doses for humans and animals, each unit containing a predetermined amount of active ingredient calculated to achieve the desired therapeutic effect, together with the required pharmaceutical diluent or carrier. The formulations for the novel unit dosage forms of the invention are determined by and depend directly on (a) the unique properties of the active material and the effect to be determined and (b) the limitations inherent in the composition of such an active agent for use in humans and animals, which are described in detail herein and constitute features of the invention. Examples of suitable unit dosage forms according to the invention are tablets, capsules, pills, suppositories, powder packs, wafers, granules, cachets, teaspoons, tablespoons, droppers, ampoules, vials, metered-release aerosols, individual multiples of one of the above doses and other forms described here.

An effective amount of the compound is used in the treatment. The dosage of the compound for treatment depends on many factors known to the person skilled in the art. For example, they include the mode of administration and the ability of the particular compound. A human dosing schedule of about 2-4000 mg of compound in a single dose, which is administered parenterally or in the compositions of the invention, is effective in treating tumors and bacterial infections. More particularly, the single dose is from 5 mg to 80 mg about 200 mg. The oral and rectal dose is between approximately 5 and 500 mg in a single dose. In particular, the dose is between about 10 and 2500 mg of compound.

Preparation 1.

5 s-ο- (3 ', 4'-di-O-Acetyl-2'-deoxy-2'-oximino-D-arabinopyranosyl) -N, N'-dicarbobenzyloxyactinin (See Scheme G).

To a solution of 7.20 g (27.10 mmol) of 3,4-di-O-acety1-2-deoxy-2-nitroso-D-arabinopyranosyl chloride in 75 ml of dimethylformamide is added a solution of 11.86 g ( 25.0 mmol) N, N'-bis-carbobenzyloxyactinamine in 50 ml dimethylformamide. The reaction mixture is stirred at room temperature under a nitrogen atmosphere for 19.5 hours. The mixture is poured into 1 l of water with vigorous stirring. The aqueous phase is decanted and the solid is taken up in 300 ml of chloroform after which the water is separated and the solvent is removed in vacuo to yield 12.6 g of a brittle white foam. The total aqueous phase is extracted three times with 100 ml portions of chloroform. The combined organics are washed with water and brine (50 ml each) and dried over sodium sulfate. The solvent is reduced to 40 ml and this solution is added to 300 ml of water with stirring. The chloroform layer is separated and the solvent removed in vacuo to yield 6.18 g of white foam, which is combined with the other portion.

Analysis of the crude product by PMR showed that most of the DMF had been removed. Chromatography on 1.25 kg of silica gel (graduated mixture of methanol and chloroform) gives 1.33 kg (1.89 mmol, 7.6%) of the pure α anomer of 5-0- (3 ', 4' -di-0-acetyl-2'-deoxy-2-oximino-D-arabinopyranosyl) -N, N'-dicarbobenzyloxy-30-actinamine, then 2.86 g of the pure β-anomer of 5-0- (3 4'-di-O-acetyl-2'-deoxy-2O-oximino-D-arabinopyranosyl) -N, N'-dicarboben-zyloxyactinamine and further 3.17 g of this β-anomer with a purity of about 90 *.

For the α anomer: IR (CHCl 3) 3550, 3400, 3070, 35 2980, 1745, 1686, 1675 (sh), 1484, 1449, 1364, 1333, 1235, 1167, 800 1 3 15 20 1026, 669 cm "1; NMR (CDCl3) ppm 7.40 (s, 10, aromatic), 6.07 (s, 1, Hj'i, 5.95 (d, J * ®3 Hz, 1, H3 '), 5 .20 (m, 7), 3.3-4.6 (m, 10), 3.12 and 3.08 (s's, 6, NCHj), 2.12 and 2.03 (s', s, 6 , CH3); MS (for tetra-trimethylsilyl derivative) 991 (M +), 689, 673: CMR (dg-acetone) ppm 5 170.1, 169.8, 157.8, 156.5, 150.1, 138, 1, 129.1, 128.8, 128.4, 91.7, 86.9, 79.1, 74.4, 69.1, 68.6, 68.3, 67.8, 67.3, 60.3, 57.7, 31.5, 31.4, 20.8, 20.5, mp 130-140 ° C, decomposition High resolution MS (tetra TMS) c gH ^ NgO ^ Si ^ calculated 991 4169 Found 991.4190.

For the β anomer: IR (CHCl3) 3500, 3100, 2970, 10 1748, 1686, 1672 (sh), 1486, 1449, 1370, 1337, 1235, 1167, 1107, 1024, 700 cm -1 NMR (CDCl 3) ) ppm 7.40 (s, 10, aromatic), 6.40 (s, 1, Hj *), 6.05 (d, Ja3 Hz, 1, H3 '), 5.0-5.5 (m, 6), 3.4-3.8 (m, 11), 3.07 and 3.04 (s's, 6, NCH3), 2.08, 2.05 (s's, 6, CH3); MS (for tetratrimethylsilyl derivative ), 991 (M +), 990, 976, 975, 689, 673; CMR 15 (dg-acetone) ppm 170.7, 169.9, 157.7, 157.1, 149.1, 137.9, 129 , 1, 128.7, 128.3, 82.3, 85.3, 74.7, 70.8, 69.4, 67.3, 60.2, 31.4, 20.8, 20.5 mp 140-155 ° C, decomposition.High resolution MS (tetra TMS) Cl2H73N3014Si ^ calculated 991.4169, found 991.4229.

Preparation 1a.

5-0- (3 ', 4', 6'-Tri-O-acetyl-2'-oximino-2'-deoxy-α-L-glucopyranosyl) - Ν, Ν'-dicarbobenzyloxyactinamine (see Scheme H).

A solution of 3,4,6-tri-0-acetyl-2-nitroso-2-deoxv-α-glucopyranosyl chloride (7.09 g, 21 mmol) in 150 ml of dimethyl-25-formaraide and N, N-dicarbobenzyloxyactinamine (14 .22 g, 30 mmol) is stirred for 18 hours until no pyranosyl chloride remains. The solution is concentrated at 45 ° C and the residue dissolved in chloroform containing 1.5% methanol. This residue is applied to a column of wet-applied silica gel (3 kg). The column is eluted with 1.5% methanol in chloroform. After using about 10 l of eluent, the main product is eluted as determined by TLC (5% methanol in chloroform). The fractions containing pure product are combined and concentrated to give 7.86 g (48%), 5-0- (3 ', 4', 6'-Tri-O-acetyl-2'-oximino-2'-deoxy-α-glucopyranosyl) -35 N, N'-dicarbobenzyloxyactinamine.

800 1 3 15 21 CD (CH OH) / ~ QJ ™ ny + 18,900 + 1,300 / “α_7ρ -53 ° (6, 0.9 acetone) NMR (CDC13): 2.02 (9H, S), 3r03 (6 .H, S) # 5.04 (4H, S), 6.20 (1H, S), 7.32 (10H, S) ppm.

5 CMR (CD3COCD3) s 170.8, 170.0, 169.8, 157.7, 157.0, 149.5, 137.9, 129.1, 128.7, 128.3, 92.0, 85.8, 74.4, 70.4, 69.8, 69.4, 68.6, 67.3, 62.5, 60.6, 60.2, 31.4, 31.1, 20, 6 ppm.

Mass spectrum, m / e (tetra TMS): 1063 (M +), 1048, 793, 792, 689, 674, 645.

10 Preparation lb.

5-0- (4 ', 6'-di-O-Acetyl-3'-O-methyl-2'-deoxy-2' -oximino-β-D-gluco-pyranosyl) -N, N'-dicarbobenzyloxyactinamine ( see diagram I).

To a solution of 60.8 g (196 mmol) of 4,6-di-O-15 acetyl-3-O-methyl-2-nitroso-α-glucopyranosyl chloride (2) in 1 L of dimethylformamide is added 97.7 g (201 mmol) Ν, Ν'-bis-carbobenzyloxy-actinamine. The reaction mixture is stirred at room temperature under a nitrogen atmosphere for 45 hours and then concentrated to a thick syrup under high vacuum at 30 ° C. It is poured in 20 parts into 3 liters of chilled water and stirred vigorously.

The white solid obtained is filtered, redissolved in 3 L chloroform, separated from the remaining water and dried over sodium sulfate. Concentration of the chloroform layer gives 143 g of a tacky white foam, which contains about 7% dimethylformamide in 25 km integral ratios.

Chromatography on 3.5 kg of silica gel using a graduated elution systeam consisting of methanol and chloroform gives a crude separation of products. The fractions containing the β anomer are crystallized in acetone to give 4.4 g of the pure β isomer. Chromatography of the mother liquors gives 1.5 more of the 5 anomer.

IR: 3500, 3310, 2920, 1750, 1690, 1459, 1240, 1175, 1105, 1403, 735, 711 cm -1.

NMR (CDCl3) 7.32s, 5.41s, 5.11s, 3.5-4.5m, 3.33s, 3.04s, 2.02s.

35 CMR (CDC13) 170.6, 169.6, 157.7, 156.5, 150.1, 136.5, 128.5, 127.8, 80 0 1 3 15 22 95.7, 92.7 , 77.1, 73.2, 70.1, 67.54, 64.09, 56.8, 29.7, 20.8, 20.5 ppm.

Mass spectrum (tetratrimethylsilyl derivative) M + 1035 mp. 214-216 ° C.

The oxime is obtained from Tables A and B by a method similar to that of Preparation 1, 1a and 1b but using the appropriately substituted actinamine and sugar reaction components.

table A

10 See Formula 14.

B Bx V V V V Ris "

- o O O

I »'* HO- HO- H- CH3COCH2- C ^ CO- H ch3 ° -

15 II II »II II

CH30- HO-

Μ H If W

C2HsO- HO- tt η η n HS- HO- "" η m ii n tl CH3S- H0-

II η II II II

CHS- HO- 20 II η Η II ·· H- HO- _ "" It H "HO- H- HO- CH30-" C-CH2- HO- HO- "BrCH2-

"II II II

HO- HS- "ClCHj- 25 HO- CH3S-" BTCH2 HO- C2H5S- "C1CH2- HO- HO-" φ CH2OCH2- <D CH2 ° "H_ <S> CH2" HO- "" CH2 ° ”" "30 »» Ho- '· "ch3o- ch3c-o-" ch3c-

HO- "" C2H5 ~ H

O O

HO- "" CH3C-OCH2- CH3C-0-CHjOCHj- 35 HO- "*" C2H5 ”800 1 3 15

TABLE A CONTINUED

23

η T} B If B Η B «o M P M

B Bj Rj R2 R3 R4 Rjs o ö

C W VI

3 HO- HO- CH3OCH2- H CH3C-0- H CH-jC- HO- "(o ^ 0Η2 ° 0Η2 ~" "" (Ó) CH2- HO- "H- H- CH30 H CH3-

O O

io ho- "" ch3c-o-ch2- ch3c-o- "ch3-

TABLE B

See formula 15 15 -

o B P p W p W Tj II p H

B Bj Rj R2 R3 R4 R1S

0 Ö Ö 1 i n HO- HO- H- CH3C0CH2- CH3C0_ H CH3C “CHjO- HO-" "" "" 20 C2H5 ° "H0_" "" ""

HS- HO- ”" Hl. II

CH3S- HO- "" M

CjHgS- HO- "" "" H- HO- "" .... o 25 HO- H- "" h HO- CH3 ° "" C1CH2- m "'· HO- HO-" BrCH2 "" HO- HS - "" "" "HO- CRjS-" "" "30 HO- C2H5S" ”ho- ho-" Qy ch2och2- 0 ch2o-h-ch2- HO- "" <ö> CH20- "" "

O O

tl ff 35 HO- "" CH3 ° "CH3C-0-" CH3C "HO-" "C2H5" H "" 800 1 3 15

TABLE B CONTINUED

24 B Bt Rj "R2" R3 "R4" Rl5 "-Ö Ö ö - H» »** S HO- HO- H- CH3C-OCH2- CH ^ -O- CT ^ OCHj CH-jC- HO-" " "C2H5" "

HO- "CH3OCH2-H" H

HO- * CH2OCH2- "" CH2- .. HO- Η- Η- O Η H CH--

10 "J

CH3OCH2-

Prepare 2 N, N'-Dicarbobenzyloxy-5-demethyl-3-O-acetyl-4- (R) -acetoxy-3 "(R) -15 dihydrospectrinomycin (see Scheme J).

5-0-0.4'-di-O-acetyl-2'-deoxy-2 '-oximino-D-arabinopyranosyl) -N, N'-dicarbobenzyloxyactinamine (2.32 g, 3.30 mmol) is dissolved in 30 ml methyl nitrile and 5.0 ml (89.4 mmol) 20 acetaldehyde and 2.5 ml (2.50 mol) 1N hydrochloric acid are added.

The reaction mixture is stirred at room temperature for 3.75 hours, sodium sulfate is added and stirring is continued for 15 minutes. The mixture is filtered and the solvent removed in vacuo to give 2.6 g of a white solid. Chromato-25 over 200 g of silica gel (mixture of methanol and chloroform) gives 1.45 g (2.11 mmol, 64%) of N, N'-dicarbobenzyloxy-5'-demethyl-3'-O-acetyl-4 ' - (R) -acetoxy -3 '- (R) -dihydrospectinomycin as a white solid: m.p. 135.0-142.9 ° C.

IR (CHCl 3) 3450, 3050, 1748, 1684, 1881, 1447, 1362, 1333, 1238, 1160, 1050, 1020, 680 cm -1.

NMR (CDCl3) ppm 7.4 (s, 10, aromatic), 3.4-5.4 (m), 3.05 (s, 6, NCH3), 2.10 (s, 3, COCH3), 1 .95 (s, 3, COCH3); CMR (d-acetone) ppm 170.2, 169.8, 157.5, 156.9, 138.0, 129.1, 128.3, 35 94.6, 91.5, 74.7, 72 , 4.67.3, 67.0, 66.3, 65.6, 61.7, 57.5, 31.6, 800 1 3 15 25 20.6; MS (for tristrimethylsilyl derivative) 904 (M +).

Preparation 2a.

See diagram K.

5 A solution of 5-0- (3 ', 4', 6'-tri-O-acetyl-2 * -oximino-2'-deoxy-α-glucopyranosyl) -N, N'-dicarbofcenzyloxyactin-amine (7, 00 g, 9.03 mmol), acetonitrile (7 ml), acetaldehyde (14.2 ml) and 1N HCl (17.0 ml) are stirred at room temperature for 5 hours. Anhydrous sodium sulfate (60 ml) is added and stirring is continued for 10 minutes. The solid is filtered and washed with acetonitrile. The filtrate is concentrated and taken up on a mixture of chloroform and ethyl acetate (1: 1 ratio) (15 ml) and chromatographed on silica gel (150 g) using the same solvent. The eluate is subjected to TLC 15 (5% methanol in chloroform) and pure fractions are combined and concentrated to yield 5.96 g (87%) hemicetal triacetate.

[mu] g7 -52 ° (C.07), chloroform.

NMR (CDCl3): 2.06 (9H, S), 2.88 (3H, S), 3.08 (3H, S), 4.90 (s), 20 5.13 (4h, S), 7 .38 ppm (10H, S).

CMR (CD3COCD3): 170.1, 169.4, 137.5, 137.4, 128.6, 127.9, 99.2, 94.0, 82.2, 73.9, 70.6, 68 , 4.68.0, 66.8, 65.6, 62.3, 60.4, 57.3, 30.0, 20.0 ppm.

Preparation 2b.

N, N'-Dicarbobenzyloxy-3'-O-methyl-4 '- (R) -6'-diacetoxy-3' - (S) -dihydrospectinomycin (see Scheme L).

To a suspension of 5-0- (4 ', 6'-di-O-acetyl-3'-0-methyl-2'-deoxy-2'-oximino-6-D-glucopyranosyl -) - N, N' -Dicarbobenzyl-30 oxyactinamine, 300 mg (0.40 mmol) in 12 ml of acetonitrile, acetaldehyde, 1.2 ml, and hydrochloric acid, 0.18 ml (1.0 N) are added and stirred for 60 hours. The homogeneous solution is stirred with sodium sulfate for 20 minutes and filtered. Chromatography of the product on 10 g of silica gel in a mixture of 15% chloroform and ethyl acetate gives 150 mg of N, N'-dicarbobenzyloxy-3'-O-methyl-4 '- (R) -6'-diacetoxy-31- ( S) - 800 1 3 15 26 dihydrospectinomycin.

NMR (CDCl3) 7.32 s, 5.1 s, 4.68 s, 3.5 s, 3.1, 2.1 ppm.

CMR (d-acetone) 171.0, 170.3, 157.8, 138.2, 129.2, 128.5, 95.8,

O

93.8, 84.5, 75.2, 73.1, 69.8, 67.4, 66.4, 65.2, 63.7, 61.2, 57.9, 5 57.5, 30 8, 29.8, 21.0, 20.7, ppm.

Mass spectrum (tritrimethylsilyl derivative) m / e (M +) 948 (M-15) 933.

Protected spectinomycin analogs from Tables C and D are obtained by a method similar to that of Preparation 2 or 2a but using the suitably substituted precursor oxime from Tables A and B.

TABLE C See formula 16 B Bl VVVV Ris "15 - o Ö II II n HO- HO- H- CH3COCH2- C ^ CO- HC ^ C- CH30- HO-" "" "C2H50- HO-" "" HS - HO- "" »" "20 ch3s- HO- C2H5S- HO-" "η» ·· H- HO- "" "" HO- H- "" "" "HO- CH30-" "n" 25 HO- C2H5 "" "" "HO- HS-" "" "HO- CH3S” "" "" "HO- C2H5S" "" "" "30 HO- HO-" <£> CH2OCH2- <Ö) CHjO- Η- CHj- HO- "" {0} CH20- »HO-" "CH.O- 0" 0 3 II I »CHgC-O-CH3C“ HO- "" C2H5 H "" 35 800 1 3 15

TABLE C CONTINUED

* y 27 η Ο η II Ο Η pH ρ II ρ Η Β Bj Rj R2 R3 R4 Rls δ 5 δ

C II II II

3 HO- HO- H- CH3C-OCH2- CH ^ -O- CB ^ OCB ^ CB ^ C- H0- I. i. «" C2H5 "" HO- "CH30CH2- Η" H "HO-" φ, CH2OCH2- ”" "(o) CH2- 0 io HO-" H CH3COCH2- "" CH3-

TABLE D

See formula 17 15

Q p pH pH pH pH pH

B Bj Rj R2 R3 R4 Rl5 0 0 0 HO- HO- H- CH3C0CH2- CE ^ CO- H CH3C "20 ch3o HO-" ”" c2h5o H0- HS- HO- ”" »" ch3s- HO- ”" "" "c2h5s- HO-" "" "25 H- HO-" "" "HO- H-" "M" "HO- CH30- H0- C2H5 ~ HO- HS-" "" "" HO- CH -S- "" "" 30 5 HO- c2H5S "" "" "" HO- HO- "CH2OCH2- CH20_ H“ CH2 ~ HO- "" <gV CH20- O 0 35 HO- "" ch3 ° - CH3C- 0- "CH3C" 800 1 3 15

TABLE D CONTINUED

28 l v V V v v ~ 5 3 HO- HO- H- C2H5 “H H" CH3C-

0 O

HO- "CH3C-OCH2- CH3C-0- CH3OCH2-" HO- "" "C2H5- HO- * CH-OCH-- Η" H "10 HO-" <ö) CH2OCH2- "" "<£> CH2 ~

O

HO- "" CH3COCH2- "" CH3 15 Preparation 2c.

N, N '-Dicarbobenzyloxy-6'-acetoxyspeetinomycin-3'-methylenol ether (see Scheme M).

N, N'-Dicarbobenzyloxy-3'-O-methyl-4 '- (R) -6-diace-20-toxy-31- (S) -dihydrospectinomycin, 101.3 mg (0.136 mmol), dissolved in 5.5 ml of acetonitrile are stirred with 350 mg of potassium bicarbonate at room temperature for 6 days. The mixture is filtered through Celite® and the bicarbonate is washed with further acetonitrile. Concentration of the filtrate gives 80.5 mg of N, N'-dicarbobenzyloxy-25'-acetoxyspectinomycin-2'-methylenol ether.

NMR (dc acetone) 7.4 s, 5.1 s, 4.8 s, 4.75 s, 4.6-4 m, 3.9 s, 3.5 s,

O

2.0 s, CMR (dg-acetone) 171.1, 158.0, 154.2, 138.2, 129.2, 128.5, 95.7, 95.5, 89.3, 75.3 , 71.7, 67.4, 66.9, 66.5, 66.2, 60.8, 60.7, 67.8, 55.6, 30.8, 20.75 ppm.

Mass spectrum (tritrimethylsilyl derivative) ra / e M + = 888, M-15, 873.

Using a method similar to that of Preparation 2a, but using suitably substituted hemiketals gives the protected spectinomycin analogs of Tables E and F.

800 1 3 15

TABLE E

See formula 18 29 tj ta o h p w p n p «B Bj Rj R2 R3 R16 5 -'- δ-

II

HO- HO- H- CH3COCH2- H CH ^ CH30- HO- "" ""

CjHgO- HO- ”" ”" HS- HO- "" "10 CH3S- HO-" .. h C2H5S- HO- "" "H- HO-" "" ”HO- H-” "" HO- CH3 ° “15 HO- C2H5-" ....

HO- HS- "" "" HO- CH3S- "" "

HO- C.H_S- "" "M

έ D

20 HO- HO- '(£) ch2och2-' · ® ch2- HO- v "@ CH20- *"

O

HO- "" ch3 ° - "ζο) C- HO-" "C2H5 ~" 25 HO- "" O CH3- ch3c-och2- HO- "" "CH3CH2" "HO-" OH H- (c} coch2 30 HO- "" "" CH2- HO- "" '· "CH3- 35 80 0 1 3 15

TABLE F

See formula 19 30 B V Y R3 "R16" 5 _

O

«I

HO- HO- H- CH3COCH2- H CH ^ CH30- HO- "" "C2H50- HO- ...." HS- HO- "" "" 10 CH3S- HO- "" C2H5S- HO- '* H- HO- "" "" HO- H- "" "n HO- CH-O-" "” "15 3 HO- C2H5" "" "" HO- HS- HO- CH3S "" "" "HO- C2H5S- "20 HO- HO-" @ CH2OCH2- "ζθ) C ^ - HO-" "CH2 °“ ""

O

HO- "" CH30- "<ö> C- HO-" "C2H5" ""

25 O

HO- "" CH3C-OCH2- CH3- HO- "" "CH3CH2-"

O

HO- "(3} COCH2- Η H- 30 HO- *" "" <g> CH2- HO- "" "" CH3 35 80 0 1 3 15 31

Preparation 3.

N, N'-Dicarbobenzyloxy-5'-demethyl vectinomycin (see Scheme N).

N, N'-Dicarbobenzyloxy-5-demethyl-3'-0-acetyl-4'-5 R-acetoxy-3 '- (R) -dihydrospectinomycin (1.32 g, 1.92 mmol) is dissolved in 20 ml acetonitrile and 1.00 g (7.24 mmol) anhydrous potassium carbonate are added. After stirring at room temperature for 20 hours, the mixture is filtered and the solid washed with an additional 20 ml of acetonitrile. The solvent is removed in vacuo, 20 ml of methanol added and the solution treated with 1.40 g (8.04 mmol) dipotassium hydrogen phosphate. The mixture is stirred at room temperature for 22 hours, filtered and the solid residue washed well with methanol. Removal of the solvent in vacuo gives 1.37 g of a gummy orange solid. Rapid filtration through 40 g of silica gel 15 (ethyl acetate) gives 950 mg of a white solid. Further purification over 2000 µl of silica gel (prepared) plates of ethyl acetate gives pure N, N · dicarbobenzyloxy-5'-demethylspectinomycin: IR (CHCl 3) 3600, 3100, 1748, 1695, 1443, 1333, 1156, 1111, 700 cm -1; NMR (CDCl 3) ppm 7.40 (s, 10, aromatic), 5.13 (s, 4), 3.1-5.0 (m), 3.05 and 2.95 (s', s) , 6, NCH3); CMR (dg-acetone) (relative to CD3cocd3) ppm 201.5, 157.1, 156.8, 137.5, 128.5, 128.2, 127.8, 97.7 , 92.4, 74.4, 74.1, 66.6, 65.8, 65.0, 60.8, 56.7, 38.2, 30.8.

By methods similar to that of Preparation 25 3, but using the suitably substituted precursor hemiketal instead of N, N-biscarbobenzvloxy-3'-O-acetyl-4 "- (R) -acetoxy dihydrospectinomycin, the protected spectino- is obtained. mycine analogues according to Tables 6 and H.

30 9Q0 13 13

TABLE G

See formula 20 32 B Βχ Rj "R2" R3 "R4" 5 -g- η

HO- HO- H- CH3COCH2- Η H

CH30- HO- "" "·" C2H50- HO- "" ....

HS- HO- "" h "10 CH-jS- HO-" "M" C2H5S- HO- "" "" H- HO- '· "" HO- H- "" "HO- CH30-" "" 15 HO- C2H5 ”" "....

HO- HS- .. h "" HO- CH3S- "" HO- C2H5S “" "" "HO- HO- CH2OCH2- <2> CH20- h- 20 HO-” "(q ± ch2 ° - h”

HO- "" ch3 ° - H

HO- "" C2H5- H

HO- "CH3OCH2-H" H

25 HO- "@ CH2OCH2-" "" 30 800 1 3 15 TABLE Η

See formula 21 33 B Bt Rj "R2'V R3" R4 "5 _— -

O

HO- HO- H- CH3COCH2- Η H

ch3o- HO- C2HsO- HO- "" 10 ES- HO- "" "" CH3S- HO- "" "c2h5s- HO- H- HO-" "" "HO- H-" "" "-. HO- CH, 0- "" "" 15 3 HO- C2H5 "" "" HO- HS- "" "" HO- ch3s “HO- C2H5S" ”" ""

20 HO- HO- * CH2OCH2- CH2 ° “H

HO- "" <@ CH20- "" HO- "" CH30- H "HO-" "C2H5 H"

HO- "CH3OCH2 H" H

HO- "@ CH2OCH2-" "" 30 800 1 3 15 34

Preparation 3a N, N-Dicarbobenzyloxy-6-hydroxyspectinomycin (see Scheme O).

To a solution of N, N-dicarbobenzyloxy-6'-5-acetoxyspectinomycin-31-methylenol ether (25 mg, 0.04 mmol) in 0.65 ml of tetrahydrofuran is added 0.5 ml of 4 N aqueous hydrochloric acid.

The solution is stirred at room temperature for 28 hours. The reaction mixture is diluted with 25 ml of chloroform and washed with 4 ml of saturated sodium bicarbonate and water, respectively. The organic layer is collected, dried over sodium sulfate, filtered and concentrated to give 22 g of N, N'-dicarbobenzyloxy-6'-hydroxy-spectinomycin.

NMR (CDCl 3): 7.25 s, 5.1 s, 4.75 s, 4.5-3.5 m, 3.0 s ppm.

CMR (CDC13): 157.5, 156.5, 136.3, 128.5, 128.1, 127.8, 96.8, 91.0, 77.0, 74.3, 74.0, 72.1, 67.6, 67.5, 65.8, 65.2, 64.4, 61.7, 57.1, 39.0, 29.7 ppm.

Preparation 3b.

N, N-Dicarbobenzyloxy-6-hydroxyspectinomycin (see Scheme P).

To a solution of N, N'-dicarbobenzyloxy-6-hydroxy-6-spectinomycin-3'-methylenol ether, 90 mg (0.143 nice), in 2.2 ml of tetrahydrofuran is added 1.6 ml of 4 N aqueous hydrochloric acid. The reaction mixture is stirred at room temperature for 24 hours and worked up as in Preparation 3a. The N, N'-dicarbobenzyloxy-6-hydroxyspectinomycin exhibits the same mobility at TLC and properties as the product of Preparation 3a. The products are combined and chromatographed on silica gel with EtOAcCH2 Cl2 (1: 1 ratio) as the elution system.

By methods similar to those of Preparations 3a and 3b, but using suitably substituted B-enol ethers instead of the B-enol ethers used in these Preparations, the protected analogs of spectinomycin are obtained from Tables I and J.

35 800 1 3 15

TABLE I

See formula 22 35 B Bj Rt H2 R3 5 -

HO- HO- H- HOCHj- H

CH30- HO- "" C2H50- HO- "" "HS- HO-" "" 10 CH3S- HO- "" "C2HgS- HO-" "" H- HO- HO- H- "HO- CH30-" " "15 HO- C2H5" "" "HO- HS-" "" HO- CH3S ~ "" "HO- C2H5S" "" "HO- HO-" <ö> CH-OCH.- 20 1 HO- "" < 5) CH20- "HO-" "CH30-" HO- * "C2H5- HO-" "HOCH2- Οη3 ° ΟΗ2 25 HO-" "" CH-

SO

HO- "CH3OCH2- Η H

HO- "CH2OCH2-" 30 800 1 3 15

TABLE J

see formula 23 36 B Bj Rj R2 1 * 3 5 —-

HO- HO- H- HOCH2- H

CH30- HO- "" "

CoH_0- HO-

Δ D

HS- HO- "" "10 CH3S- HO-" ""

CjHgS- HO- "Η- HO- HO- H-" "" HO- ch3 ° - "" "15 HO- C2H5" "" "HO- HS- HO- CH3S- HO- C2H5S" HO- HO- "CH2OCH2 - 20 HO- "" <g> ch2o- HO- "" * CH30- "HO-" "C2H5" ho- "" hoch2- ch3och2 25 H0 "" "" C2H5

HO- "CH30CH2- Η H

HO- "<§> CH2OCH2- 30 800 1 3 15 37

Preparation 3c.

N, N * -Dicarbobenzyloxy-2'-O-acetyl-4 ', 5'-didehydrospectinomycin (see Scheme Q).

Anhydrous potassium bicarbonate (14.58 g, 146 meq.) And acetonitrile (384 ml) are heated in a flask equipped with a distillation head. After 40 ml of solvent have been distilled, the remaining suspension is cooled to room temperature under nitrogen and then hemiketal triacetate from Preparation 2a is added. The mixture is stirred at room temperature for 41 hours, after which, as shown by TLC (5% methanol in chloroform), the starting material is gone. The solid is filtered, washed and the filtrate concentrated. The residue is taken up in chloroform (40 ml) and the inorganic substances are filtered. The filtrate is concentrated to give a foam (14.51 g). After dissolving the foam in chloroform and cooling, a mass (7.55 g) is formed, which is recrystallized from chloroform (20 ml) and 3.42 g of N, N'-dicarbobenzyloxy-2'-O-acetyl-4 ' 5'-didehydrospectinomycin, mp 149-152 ° C. Chromatography of the combined mother liquors on silica gel (800 g) using a mixture of methanol and chloroform (ratio 1: 4) gives 3.37 g of product (total yield 55%).

UV (CjHgOH): 204.5 nm (20,950), 206 sh (20650), 267 (11,550), CD (CH3OH) / "\ 7 ^ o -22,000, /" \ 7 ^ +33,500, -43 ° (C , 1.25 acetone).

NMR (CDCl 3): 2.07 (3H, s), 2.15 (3H, S), 3.08 (6H, s), 5.12 (4H), 5.40 (1H, s), 5, 95 (1H, s), 7.32 (10H, s).

CMR (CD3COCD3): 182.5, 172.9, 169.5, 137.6, 137.5, 128.6, 127.8, 102.7, 95.5, 93.1, 75.0, 74 0.66.8, 65.6, 60.3, 59.6, 56.7, 31.1, 20.4 ppm.

Exact mass: 784.3097: Calculated for ^ 39 ^ 52 ^ 2 ^ 12 ^ 84.3059.

Using a method similar to that of Preparation 3, but using the suitably substituted hemiketal instead of the hemiketal prepared in Preparation 2a, the protected didehydrospectinomycin analogs of Tables K and L are obtained.

800 1 3 15 38

TABLE K

See formula 24 B Bj Rj r15 5 -

O

HO- HO- CH3- CH3C "

O

vt CH30- HO- CH3- CH3C "10 ° C2H50- HO- CH3- CH3C-

O

II

HS- HO- CH3- CH3C- 0 tf CH-S- HO- CH, CH, C- 15 3 3—3

O

C2H5S- HO- CH3- CH3C “

O

H- HO- CH3- CH3C "20 ° HO- H- CH3- CH3C"

O

II

HO- CH3 ° "CH3 ~ CH3C- 0 25 H °" C2H5 'CH3- CH ^ C- 0 tl HO- H2- CH3- CH3C ”

O

II

HO- CH3S- CH3- CH3C "30?, HO- C2H5S" CH3_ CH3CH2C- 0 tl HO- HO- CH3- CH3CH 2C- 0

Vt 35 HO- HO- CH30-CH (CH3) - CH3 (CH2) 2C- 800 1 3 15

TABLE K CONTINUED

B Βχ Rt R15 39

O

5 HO- HO- CH3OCH2-CH3 (CH2) 2C-0

II

HO- HO- C2H5 CH3 (CH2) 3C-

O

0 HO- HO- CH3- CH3 (CH2) 3C- HO- HO- CH3- isopropionyl HO- HO- C3H7 "sec.butyryl HO- HO- C3H7" tert.butyryl

15 TABLE L

See formula 25 B Bj Rj R15

20 O

II

HO- HO- CH3- CH3C-

O

I »CH30- HO- CH3- CH3C"

O

tp 25 C 2 H 5 ° “h ° - CH 3 -CH 3 C- 0

II

HS- HO- CH3- CH3C-

O

II

CHjS- HO- CH3- CH3C "

30 O

C2H5S- HO- CH3- CH3C “

O

II

H- HO- CH3- ch3c-

O

II

35 HO- H-CH3-CH3C- 800 1 3 15 40

TABLE L CONTINUED

B Bj Rj Rj ,.

5 ° “HO- CH3 °“ CH3 ”CH3C- 0 lt HO- C-H, .- CH, - CH, C- 2 3 3 3

O

10 H0- V CH3- CH3J "

O

II

HO- CH3S- CH3- CH3C "

O

«I

HO- C-H-S-CH, - CH.CH.C- 2 5 3 3 2 15? HO- HO- CH3- CH3CH2C- 0 HO- HO- CH30-CH (CH3) - CH3 (CH2) 2C- 0 tl 20 HO- HO- CH3OCH2- CH3 (CH2) 2C-

O

II

HO- HO- C2H5 "CH3 (CH2) 3C-

O

II

HO- HO- CH3- CH3 (CH2) 3C- 25 HO- HO- CH3- isopropionyl HO- HO- C3H7 "sec.butyryl HO- HO- C3H7" tert.butyryl 30 80 0 1 3 15 41

Preparation 4 N, N'-Bi scarbobenzyloxy-4 ', 5'-didehydrospectinomycin (see Scheme R) N, N'-Biscarbobenzyloxy-2'-O-acetyl-4,51-didehy-5 drospectinomycin (1.0 g) is added to a suspension of dipotassium hydrogen phosphate (0.40 g) in anhydrous methanol (20 ml) and stirred at room temperature for 1½ hours. The solvent is removed under reduced pressure and the organics are dissolved in 1% methanol in chloroform. The product-containing fractions are combined and concentrated to yield 0.51 g (55%) of Ν, Ν-biscarbo-benzyloxy-4 ', 51-didehydrospectinomycin.

CD (CH3OH): Z-®L? 314 _8 »300 ± 2,100 / ^ 7 ^ 6 + 10 * 500 ± 2 * 100 Z \ 7D

-56 ° (C 1.0, CH 3 OH).

CMR (CD3COCD3) j 187.6, 175.8, 157.2, 138.1, 128.4, 101.7, 99.3, 87.7, 76.3, 64.6, 63.8, 67.3, 66.7, 66.3, 65.3, 60.8, 60.0, 31.5, 21.3 ppm.

Mass spectrum: m / e (triTMS)? 814 (M +), 799 (M-15).

The protected didehydrospectinomycin is obtained by a method similar to that of Preparation 4, but using the suitably substituted didehydrospectinomycin derivative instead of Ν, Ν'-biscarbobenzyl-oxy-2'-O-acetyl-4 ', 5'-didehydrospectinomycin. analogues from Tables M and N.

TABLE M See formula 26 25 _____ B Bj Rj HO- HO- CH3 ~ CH30- HO- CH3- 30 C2H5 ° "h ° - CH3- HS- HO- CH3- CH3S- HO- CH3- c2h5s- bo- ch3- H - BO- CH-- 35 BO- H- CB-- 800 1 3 15

TABLE · M CONTINUED

B Βχ Rj 42 5 HO- CH30- CH3- HO- C2H5- CH3- HO- H2- CH3- HO- CH3S ”CH3” HO- C2H5S ”CH3“ 10 HO- HO- CH3- HO- HO- CH3- HO- HO- CH3OCH2- HO- HO- C2H5 “HO- HO- CH3- 15 HO- HO- CH3- HO- HO- C3H7 TABLE N See formula 27 20 B Bj Rj HO- HO- CH3- 25 CH30- HO- CH3- C2HsO- HO- CH3- HS- HO- CH3- CHjS- HO- CH3- C2H5S- HO- CH3- 30 H- HO- CH3- HO- H- CH3- HO- CH30- CH3- HO- C2H5- CH3- HO - H2- CH3- 35 HO- CH3S ”CH3 800 1 3 15

TABLE N CONTINUED

43 B Bj Rj 5 HO- C2H5S "CH3" HO- HO- CH - HO- HO- CH3- HO- HO- CH ^^ OCH ^ - HO- HO- C2H5 "10 HO- HO- CH3- HO- HO- CH3- HO- HO- C3H? -

Preparation 5 N, N'-Dicarbobenzyloxy-6'-hydroxyspectinomycin-3'-methylenol (see Scheme S).

The enol ether N, N'-dicarbobenzyloxy-5'-acetoxy-spectinomycin, 3'-methylenol ether, 103.9 mg (0.154 mmol), dissolved in 6.8 ml of methanol, is stirred with potassium bicarbonate for 24 hours at room temperature. The mixture is concentrated, and the residue is dissolved in chloroform, filtered and concentrated again to give 90 mg of N, N'-dicarbobenzyloxy-6'-hydroxyspectinomycin, 3'-methylenol ether. No methyl-25 acetate is observed at NMR and CMR.

NMR (CDCl3) j 7.31 s, 5.10 s, 5-3.75 m, 3.5 s, 3.1 s ppm.

CMR (CDC13): 158, 152.5, 136.5, 128.2, 95.1, 94.9, 88.7, 77.2, 67.5, 67.2, 55.0, 34.1 , 29.72 ppm.

Mass spectrum: (tetra-trimethylsilyl derivative) m / e (M +) - 918 (M-15) 30 903.

The protected spectinomycin analogs from Tables O and P are obtained by a method similar to that of Preparation 5, but using the suitably substituted enol ethers.

35 800 1 3 15 B Βχ Rx R2 44

TABLE O

See formula 28 5 - HO- HO- H- HOCH2- CH30- HO- "" C2H50- HO- "" HS- HO- "" 10 CH3S "HO- C2HsS- HO-" H- HO- "HO- H- "" HO- CH30- "” 15 HO- C2H5- HO- HS- "" HO- CH3S- HO- C2H5S ”20

TABLE P

See formula 29

B Bl V Y

25 HO- HO- H- HOCH2- CH30- HO- "" C2H50- HO- "HS- HO-" CH-S- HO- "30 3 C2H5S- HO-" "H- HO- H" HO- H- "" HO- ch3 ° - 35 H ° - C2H5- 800 1 3 15

TABLE P CONTINUED

45

B Bt V V

5 HO- HS- H- HOCHj- HO- CH3S- HO- C2H5S ~

Example I

Spectinomycin (see Schedule T).

To a solution of 480 mg (0.80 mmol) N, N'-biscarbobenzvloxy-4 ', 5'-didehydrospectinomycin in 40 ml isopropanol, 480 mg 10% palladium on barium sulfate and 0.48 ml (5.9 mmol) 15 pyridine added. The mixture is hydrogenated at normal pressure for 5 hours, filtered and the filtrate concentrated in vacuo. The residue is redissolved in isopropanol and treated with 2.0 ml (2.0 mmol) of an IN solution of hydrogen chloride in isopropanol. Removal of solvent in vacuo gives 440 mg of solid. Re-20 crystallization from aqueous acetone gives 164 mg (0.33 mmol, 41%) spectinomycin dihydrochloride pentahydrate with the same physical and biological properties as the natural product.

Using the same method on the enone acetate prepared in Preparation 3c, a 35% yield of crystalline spectinomycin dihydrochloride pentahydrate is obtained. Example II

5-Desmethylspectinomycin (see Scheme U).

A solution of 333 mg (0.57 mmol) Ν, Ν'-dicar-30 bobenzyloxy-5'-desmethylspectinomycin in 5 ml isopropanol is added in a Parr bottle to 300 mg 10% Pd on carbon in 45 ml isopropanol. The solution is treated with 1.36 ml (1.36 mmol) 1N hydrogen chloride in isopropanol and hydrogenated at room temperature for 16 hours at a pressure of 2.8 at. The material is filtered and the catalyst washed with isopropanol and then with water.

800 1 3 15 46

The product is concentrated in vacuo to dryness and the water-soluble portion is separated. Removal of the solvent in vacuo leaves a white residue, which is recrystallized from aqueous acetone to give 112 mg of 5'-desmethylspectinomycin 5 as a white solid.

Mp. 196-205 ° C decomposition; MS m / e (penta TMS derivative) 687.3378; calculated for C 1 H N N Sig 678.3403; CMR (D30) (refers to external TMS) ppm 95.3, 94.9, 93.6, 71.1, 67.4, 67.0, 62.9, 62.5, 61.0, 59, 9, 35.9, 32.2, 31.7.

Example III

6'-Hydroxyspectinomycin (see Scheme V).

36 mg of N, N'-Dicarbobenzyloxy-6-hydroxyspectinomycine is shaken with 20 mg of palladium black in 35 ml of isopropanol under a hydrogen pressure of 0.28 at. The catalyst is filtered and washed with isopropanol. The filtrate is concentrated to a total volume of 10 ml after which 0.12 ml of 1.0 N hydrochloric acid in isopropanol is added. Concentration of the solution gives a white residue, which is redissolved in DIO for 13c determination. isolating 20 of the 13C sample gives 14 mg of material after lyophilization. This is crystallized from water and acetone to give 6'-hydroxyspectinomycin.

Snqat. 201-205 ° C (dec.).

CMR (D20) 94.9, 93.2, 73.6, 70.7, 67.2, 64.6, 62.7, 60.8, 59.6, 37.0, 31.9, 31 , 4 ppm.

Spectinomycin Rf. 0.5.

6'-Hydroxyspectinomycin Rf; 0.25,

The crude product 61-hydroxyspectinomycin shows bactericidal activity.

Using methods similar to those of Examples I, II and III, but using the appropriately substituted protected spectinomycin instead of N, N * bis-carbobenzyloxy-4 ', 5'-didehydrospectinomycin, N, N'-bis-carbobenzyloxy-2 "-O-acetyl-41,5'-didehydrospectinomycin and N, N'-dicarbobenzyloxy-5'-desmethylspec-35 tinomycin and N, N'-dicarbobenzyloxy-6'-hydroxyspectinomycin 800 1 3 15 47 are obtained in the tables Q and R indicated spectinomycin analogs.

TABLE Q

See formula 30 5 ______ B B1 * 2 HO- HO- H- HOCH2- CH30- HO- H- H0CH2- 10 C2H5 ° "HO- H- HOCH2- HS- HO- H- HOCH2- CH3S- HO- H- HOCH2 -

CjHgS- HO- H- HOCH2- H- HO- H- HOCH2- 15 HO- H- H- HOCH2- HO- CH3 ° - H- HOCH2- HO- C2H5 “H“ HOCH2- HO- H- H- H0CH2 " HO- CH3S "H" HOCH2- 20 HO- C2H5S ~ H “HOCH2- HO- HO- H- HOCH2- HO- HO- H- C2H5” HO- HO- H- HOCH2- HO- HO- H- HOCH2- 25 HO- HO- CH3OCH2- H- HO- HO- HOCH2- H- HO- HO- H- CH3- 30 800 1 3 15 48 TABLE R See formula 31 B Bj R2 5 --—-------— -— HO- HO- H- HOCHj- CH30- HO- H- HOCH2- C2H50- HO- H- HOCH2- HS- HO- H- HOCH2- 10 CH3S- HO- H- HOCH2- C2H5S- HO- H- HOCH2 - H- HO- H- HOCHj- HO- H- H- HOCH2- HO- CH3 ° - H- HOCH2- 15 HO- C2H5- H- HOCH2- HO- H- H- HOCH2- HO- CH3S "H" HOCH2 - HO- C2H5S "H" HOCHj- HO- HO- H- HOCH2- 20 HO- HO- H- C2H5 ~ HO- HO- H- HOCH2- HO- HO- H- HOCH2- HO- HO- CHgOCHj- H- HO- HO- HOCHj- H- 25 HO- HO- H- CH3-

Example IV

4 ', 51-Didehydrospectinomycin dihydrochloride (see Scheme W).

A mixture containing N, N'-biscarbobenzyloxy-4 ', 5 * - didehydrospectinomycin (93 mg), 10% palladium on carbon (70 mg), isopropanol (30 ml) and 0.1 N HCl in isopropanol (2 ml ) is hydrogenated for 1½ hours at a pressure of 2.1 at. At this point further catalyst (30 mg) and 0.1 N HCl in isopropanol (1.0 ml) are added and hydrogenation is resumed for another 2½ hours. Then the solid is filtered off and the filtrate concentrated to give 50 mg of 4 ', 5'-didehydrospectinomycin dihydrochloride as a solid. This product is active against E coli and does not contain spectinomycin or dihydrospectinomycin, as shown by 5 different DLC systems (CHCl 3: CH 2 OH: NH 4 OH, ratio 3: 4: 2) (3% NH 4 OH in acetone). In this system, it exhibits a pv active spot, corresponding to a reference standard of enantiomer 41,5'-dehydrospectinomycin, which is characterized as follows:

Mass spectrum (tri-TMS): m / e 546 (M +), 531 (H-15), 10 492, 426, 401, 361, 271, 199.

CMR (DjO external TMS): 189.6, 179.3, 102.3, 98.5, 72.6, 69.9, 67.0, 66.5, 62.6, 61.5, 59.7 , 32, 32.1, 22.0 ppm.

Using a method similar to that of Example IV but using the suitably substituted 4 ', 5'-didehydrospectinomycin instead of N, N'-biscarbobenzyloxy-4', 51-didehydrospectinoraycine, the 4 ', 5 * -didehydrospec is obtained -tinomycins from Tables S and T.

TABLE S See formula 32 20______ B Bj Rj HO- HO- CH3- CH30- HO- CH3- 25 C2H5 ° “HO- CH3 ~ HS- HO- CH3- CH3S- HO- CH3- C2H5S- HO- CH3- H- HO - CH3- 30 HO- H- CH3- HO- CH3 ° “CH3 ~ HO- C2H5- CH3- H0- H2- CH3- HO- CH3S- CH3- 35 HO- C2H5S" CH3 “800 1 3 15

TABLE S CONTINUED

50 B Bj Rj 5 HO- HO- CH3- HO- HO- C2H5 ”HO- HO- HOCHj- HO- HO- HOCH2- HO- HO- C3H7- 10 HO- HO- C3H3"

TABLE T

See formula 33 15 --- BB, R, 1 1 HO- HO- CH3- CH30- HO- CH3- C_H_0- HO- CH-- 20 2 5 3 HS- HO- CH3- CH3S- HO- CH3- C2H5S- HO- CH3- H- HO- CH3- 25 H ° - H 'CH3 "HO- ch3 ° - CH3- HO- C2H5" CH3 ~ HO- H2- CH3- HO- CH3S- CH3- 30 HO- C2H5S- CH3- HO- HO- CH3- HO- HO- C2H5 "HO- HO- HOCH2- HO- HO- HOCH2- 35 HO- HO- C3H7" HO- HO- C3h3- 80 0 1 3 15

Claims (42)

Anomers and asteric mixtures of a compound characterized by the formula 1, wherein R represents a group of the formula 8 or 9 where-5 in R 1 to R 1 -ad may be or not equal and a hydrogen atom, lower alkyl, lower alkenyl, lower haloalkyl, low aminoalkyl, lower alkynyl, -ox or - (CH2) ^ - OX and the isomer and represent thereof, with the proviso that R1 and R2 are not a hydroxy group and that one of the groups Rg and Rj must be hydrogen, 10. is a hydrogen atom, lower alkyl, lower alkenyl or lower alkynyl group; η is 1, 2, 3 or 4, Rj. a hydrogen atom or lower alkyl group, Rc to R4. a hydrogen atom, lower alkyl, lower o 14 15 alkenyl or lower alkynyl group, with the proviso that one of the groups R 1 and Rg and one of the groups and RJ2 is always hydrogen and B and are or are not equal and represent a hydrogen atom, hydroxyl, alkoxy, o-lower alkenyl, thio, thio lower alkyl-20 or thio lower alkenyl group and A is an oxygen or sulfur atom with the proviso that when the compound of formula 10 cannot be a hydroxyl group, hydrated forms thereof and pharmaceutically acceptable salts of the compound of formula 1 and hydration standards thereof. A compound according to claim 1, characterized by the formula Ib, wherein R 1 to R 1, A, B and have the meaning indicated in claim 1. 3. A compound according to claim 2, characterized by the formula 34, wherein Rg and R1 represent a lower alkyl group and B and Bj represent a hydroxyl group or lower alkoxy group. Compound according to claim 3, characterized in that it is 5-desmethylspectinomycin. 5 R1 is a hydrogen atom or lower alkyl group, R'7, R'g, R'and R'12 represent a lower alkyl, lower alkenyl, lower alkynyl, protecting aralkoxycarbonyl, halogenated alkoxycarbonyl or alkoxycarbonyl group with the proviso that one of the groups R'7 and R'g and one of the groups A compound according to claim 2, characterized by the formula 35. 800 1 3 15 A compound according to claim 1 / characterized by the formula 60, wherein R 1 to R 1, A, B and Bj have the meaning as defined in claim 1. 7. A compound according to claim 6, characterized by the formula 36, wherein R 1, R 8 and R 1 represent a lower alkyl group and B and B 1 represent a hydroxyl or alkoxy group. A compound according to claim 7, characterized in that it is 4 ', 5'-didehydrospectinomycin. 9. Dihydrochloride salt of the compound according to claim 7. R '?, R'g, R' and R'12 have the meaning indicated in claim 29. R'jj and R'12 is always a protecting group, Rjg is a hydrogen atom or acyl group, A is an oxygen or sulfur atom and B and a hydrogen atom, hydroxyl, alkoxy, o-lower alkenyl, thio, thio lower alkyl or thio lower alkenyl group 15 with the proviso that when the compound has the formula 40 wherein CB 2 is a carbobenzyloxy group it cannot be hydrogen or hydroxyl group. 10. Process for the preparation of anomers and asteric mixtures of a compound of the formula 37, characterized in that a) a compound of the formula 61 is converted into a compound of the formula 2 'and b) the protected group (and ) removes from the compound formed in step a) wherein R is a group of the formula 8 or 9 wherein R 1 to m are equal or not and a hydrogen atom, lower alkyl, lower haloalkyl, lower aminoalkyl, lower alkenyl - a lower alkynyl group, -ox or - (CH_) ox and isomers thereof, with the proviso that R ^ and R ^ do not represent a hydroxy group and one of the groups Rg or R ^ must be hydrogen, X hydrogen atom, lower alkyl, lower alkenyl or lower alkynyl group, η is 1, 2, 3, or 4, R5 is a hydrogen atom or lower alkyl group, Rg to Rj ^ is a hydrogen atom, lower alkyl, lower alkenyl or lower alkynyl group; 11. Process according to claim 10, characterized in that the compound prepared has the formula 2b, wherein Rj to ® y »^ 10f R'll 'R'i2' R13 # Rj4 * ® 10 have the meaning indicated in claim 10 . 12. A process according to claim 11, characterized in that the compound prepared has the formula 38 wherein R'g and R * 2 represent a lower alkyl group and B 'and B' 2 represent a hydroxyl or lower alkoxy group. The method according to claim 12, characterized in that the compound prepared is 5-deoethyl vectinomycin. Process according to claim 10, characterized in that the compound prepared has the formula 62, wherein Rj, A, B and Bj have the meaning indicated in claim 10. Process according to claim 14, characterized in that the compound prepared has the formula 39, wherein R 1, R 8 and R 12 represent a lower alkyl group and B and B 1 represent a hydroxyl or alkoxy group. 16. A method according to claim 15, characterized in that the compound prepared is 4 ', 5'-didehydrospectinomycin, 17, Anomers and asteric mixtures of a compound characterized by the formula 2, wherein R represents a group of the formula 8 or 9, wherein R 1 to R 2 may be the same or different and a hydrogen-30 atom, lower alkyl , lower haloalkyl, low aminoalkyl, low alkenyl, lower alkynyl, -ox or - (cell) -ox, with the proviso that R 1 and R 2 are not a hydroxyl group and R 1 and R 1 must be hydrogen, X a hydrogen atom lower alkyl, lower alkenyl-35 or lower alkynyl group, 800 is 1 3 15 η 1, 2f 3 or 4, R 1. a hydrogen atom or lower alkyl group, Rg, Rg, R10 # R13 and R14 represent a hydrogen atom, lower alkyl, lower alkenyl or lower alkynyl group, Compound according to claim 17, characterized by the formula 64, wherein Rj to r ^, Rg, R 'R'g, Rg, R ^ g, R' A compound according to claim 18, characterized by the formula 41, wherein R'g and R'J2 represent a lower alkyl group, B represents a hydrogen atom or lower alkoxy group and 20. A compound according to claim 19, characterized in that it is N, N'-biscarbobenzyloxy-5-desmethylspectinomycin. R'12 'R13' R14 * B and Bj have the meaning indicated in claim 17. A compound according to claim 18, characterized by the formula 42, wherein CBz is a carbobenzyloxy group. A compound according to claim 17, characterized by the formula 63, wherein V V RV RV V V R, 11 'R' 12 'r,', R. ', A, B and B have the meaning as defined in claim 17. 13 14 1 Compound according to claim 22, characterized in that RJ5 is hydrogen. 24. A compound according to claim 23, characterized by the formula 43, wherein R 1, R'g and R '' are a lower alkyl group, A is oxygen or sulfur and B represents a hydroxyl or alkoxy group. Rg is a hydrogen atom or a lower alkyl group, Rg, Rg, Rj0 # Rjg, and R ^ are a hydrogen atom, lower alkyl, lower alkenyl or lower alkynyl group, Rg is a hydrogen atom or alkyl group, R ', R'g, R'jj and R'12 represent a lower alkyl, lower alkenyl, lower alkynyl, protective aralkoxycarbonyl, halogenated alkoxycarbonyl or alkoxycarbonyl group, with the proviso that one of groups R * 7 and R'g and one of the groups R'11 ^ R'12 is a protecting group, R1 is a lower alkyl, aralkyl or aroyl group, 1635. is an oxygen or sulfur atom and 800 1 3 15 B and Bj al or are not the same and represent a hydrogen atom, hydroxyl, alkoxy, o-lower alkenyl, thio, thio-lower alkyl or thio-lower alkenyl group. 25. A compound according to claim 24, characterized by the formula 44. CBz is a carbobenzyloxy group. A compound according to claim 22, characterized in that R 15 is an acyl group. A compound according to claim 26, characterized by the formula 45, wherein R ', R'g and R'12 are a lower alkyl group, 10. and Bj represent a hydroxyl or alkoxy group and Ac is an acyl group. 28. A compound according to claim 27, characterized in that it is N, N, -dicarbobenzyloxy-2'-o-acetyl-4 ', 5'-didehydro-spectinomycin. 29. Anomers and asteric mixtures of a compound, characterized by the formula 46, wherein R 1 to R 6 may or may not be the same and a hydrogen atom, low alkyl, low haloalkyl, low aminoalkyl, low alkenyl, low alkynyl group, -OX or - (CH.) ox and its isomers thereof, with the proviso that R1, R2 and R8 are not a hydroxyl group, wherein X is a hydrogen atom, lower alkyl, lower alkenyl or lower alkynyl group is, η is 1, 2, 3 or 4, A compound according to claim 29, characterized by the formula 47, wherein RJf R2, Rg, Rg, Rg, RJ0, RJ3, R1 (J, Rjg, R ', R', R ', and R', B and B have the meaning indicated in claim 29. Rg is hydrogen or a low alley group, R'7, R'q, R'jj and R 'J2 is a lower alkyl, lower alkenyl, lower alkynyl, a protective aralkoxycarbonyl, halogenated alkoxycarbonyl or alkoxycarbonyl group proposing, with the proviso that one of the groups R 'and R' and one of the 7 or 35 groups R '^ and R' ^ is always hydrogen and further, with the proviso that one of the groups R '^ and R'g and one of the groups R'jj and R' is always a protecting group, A is an oxygen or sulfur atom and B and Bj are the same or different and a hydrogen-5 atom, hydroxyl, alkoxy -, o-lower alkenyl, thio, thio lower alkyl, or thio lower alkenyl group. A compound according to claim 30, characterized by the formula 48, wherein Rg is a lower alkyl group and R 1, R 2, R 3, A compound according to claim 31, characterized by the formula 49, wherein R 1, R 2, R 3 and R 16 have the meaning as defined in claim 29 and CB 2 is a carbobenzyloxy group. A compound according to claim 32, characterized by the formula 50, wherein R 2 and CB 2 have the meaning as defined in claim 32. A compound according to claim 33, characterized by the formula 51, wherein CB 2 is a carbobenzyloxy group. 35. Process for preparing anomers and asteric mixtures of a compound characterized by the formula 3b by converting a) a compound of the formula 52 into a compound of the formula 53 and 25 b) converting the compound from step a) into the compound of the formula 3b wherein R 1 to R 3 may or may not be the same and represent a hydrogen atom, lower alkyl, lower haloalkyl, lower aminoalkyl, lower alkenyl, lower alkynyl, -OX or - (CEL) ox with the proviso that R1, R2 and R3 do not represent a hydroxy group, wherein X is a hydrogen atom, a lower alkyl, lower alkenyl or lower alkynyl group, η is 1, 2, 3, or 4, Rg is a hydrogen atom or lower alkyl group, 35 and R 1. hydrogen, a lower alkyl, 6 y 10 13 14 800 1 3 15 lower alkenyl or lower alkynyl group, Rj. a hydrogen atom or lower alkyl group, R ', R'g, R'jj and R'12 represent a lower alkyl, lower alkenyl, lower alkynyl, protective aralkoxycarbonyl, halogenated alkoxycycarbonyl, or alkoxycarbonyl group, with the proviso that one of the groups R'7 and R'g and one of the groups R '^ and R'^ is always a protecting group, Rjg is a lower alkyl, aralkyl or aroyl group, A oxygen or sulfur and 10. and Bj is hydrogen, a hydroxyl, alkoxy, o-lower alkenyl, thio, thio-lower alkyl or thio-lower alkenyl group. A method according to claim 35, characterized in that the compound prepared has the formula 54, wherein R 1, R 1, 15 β 3. V R '?, 8s, 83, B10> B'u. R'u, R13, R14, R16, B and Bt have the meaning indicated in claim 35. 37. Process according to claim 36, characterized in that the compound prepared has the formula 55, in which R16 is a lower alkyl group and R1, R1, R1, R'7, R'g, r'h 611 R'12 Claim 36 have the meaning indicated. 38. Process according to claim 37, characterized in that the compound prepared has the formula 56, wherein R 1, R 1, R and R 1 have the meaning as defined in claim 36 and CB 2 is a carbobenzyloxy group. A method according to claim 38, characterized in that the compound prepared has the formula 57, wherein and CB 2 have the meaning indicated in claim 37. 40. A process according to claim 39, characterized in that the compound prepared has the formula 58, wherein CB 2 is a carbobenzoyloxy group. 41. Pharmaceutical preparations with antibiotic activity, characterized in that the preparation as active ingredient contains an anomeric or asteric mixture as defined in claims 1-9, alternatively as prepared according to the method according to claims 10-40. 800 1 3 15 5 42. Compounds and preparations as described in the description and the examples. 800 1 3 15