HU181535B - Process for preparing new analogues of spectinomycin - Google Patents

Abstract

An anomer or an asteric mixture of a compound of the formula <IMAGE> wherein R is selected from <IMAGE> wherein R1 to R4 may be the same or different and are selected from hydrogen, lower alkyl, lower alkenyl, lower haloalkyl, lower aminoalkyl, lower alkynyl, -OX and -(CH2)n-OX and isomers thereof with the provisos that R1 and R2 are not hydroxy and one of R3 and R4 must be hydrogen, wherein X is selected from hydrogen, lower alkyl, lower alkenyl, and lower alkynyl, n is an integer of from one to four, R5 is hydrogen or lower alkyl; R6 to R14 are selected from hydrogen, lower alkyl, lower alkenyl and lower alkynyl with the proviso that one of R7 and R8 is always hydrogen and one of R11 and R12 is always hydrogen, and B and B1 are the same or different and are selected from hydrogen, hydroxy, alkoxy, o-lower alkyenyl, thio, thio- lower alkyl and thio-lower alkenyl, and A is oxygen or sulfur; or a hydrate or a pharmaceutically acceptable salt or salt hydrate thereof; excluding spectinomycin, are novel compounds. These novel compounds, and also spectinomycin, are prepared by a synthetic route involving coupling of an actinamide with a sugar. They can be used for treating bacterial infections and tumours.

Description

This invention relates to novel analogs of spectinomycin.

It is known that spectinomycin (VIII) has been prepared microbiologically by the method described in U.S. Patent No. 3,234,092.

Rosenbrook Jr. et al., J. Antibiotics, 1975, 28, 953-960 and ibid, 31, 451 (1978) disclose some analogs of spectinomycin. Camey et al., J. Antibiotics, 1977, 30, 960 (1977) describe chlorodeoxy derivatives of spectinomycin. Finally, Foley et al., J. Org. Chem., 43 (22) 4355-4359 (1978) discloses 9-epi-4 (R) -dihydro-spectinomycin.

None of the aforementioned literature discloses that any analog or derivative of spectinomycin is biologically active.

The method which is closest to the process of the invention described below is described by Lemieux in Can. J. Chem., 51.53 (1973). According to this method, the 5-hydroxy group of the 2-deoxy-streptamine of the formula (I) reacts preferentially with the tri-O-acetyl-2-deoxy-2-nitroso-α-D-glucopyranosyl chloride of the formula (2). pseudodisaccharide (see Scheme A wherein CBz represents a benzyloxycarbonyl group). Mallams et al., J. Chem. Soc. Perkin, I. 1118 (1976), extended the Lemieux method for the synthesis of di- and trisaccharides.

The conversion of oximes is described by Lemieux on page 19 of the abovementioned publication 181535 and Mallams et al. On page 1097 of the abovementioned reference.

It has been found that the novel analogues of spectinomycin (I) have valuable biological activity. In the general formula (I)

R is (X) or (IX)

R 1 is hydrogen or C 1-4 alkyl, halo-C 4 alkyl, or hydroxy (C 1-4) alkyl as defined below, but other than methyl;

R 1 is C 1-4 alkyl,

R ₂ and R ₃ are each independently alkyl of 1 to 4 carbon atoms.

The numbering of carbon atoms in formula (I) as set forth below will be used throughout the specification.

Pharmaceutically acceptable salts of the compounds of formula (I) may also be prepared according to the process of the invention.

In addition to the description of substituents of formula I, the alkyl group includes methyl, ethyl, propyl or butyl groups, and isomeric forms thereof.

The term "halogen-C4 alkyl" in the alkyl portion of the above-mentioned alkyl group refers to any one of two alkyl groups substituted by one, two or three halogens.

By "halogen" is meant fluorine, chlorine, bromine or iodine.

It has further been found that the stereochemistry of the glycoside bond, i.e. the Γ-position of the compounds of formula (I)

-1181535 is extremely important as the β-anomer is more biologically active.

Thus, the term "α-anomer" means that a substituent at the Γ position is located below the plane of the ring system, while the term "β-anomer" means that the substituent at the position Γ is located above the plane of the ring system. The configuration of spectinomycin corresponds to that of the β-anomers.

The β-anomers exhibit desirable biological activity. Therefore, in order to produce biologically active analogues of spectinomycin, sufficient selectivity for the Γ-position must be ensured.

, Actinic amines and actinin derivatives include aminocyclitols of formula VI.

The decisive advantage of the process of the present invention, which will be described below, is the ability to selectively produce antimicrobial agents which are analogs of spectinomycin in a complete chemical synthesis. To date, it has not been recognized that a variety of spectinomycin analogues can be prepared by a process that produces a biologically active six-membered hemiketal having a structure similar to spectinomycin by converting delta-hydroxy oximes into delta-hydroxy ketones.

A further advantage of the process according to the invention is that the actinines bearing the protected amino groups used as starting material react at their 5-hydroxyl group without the need to protect any further β- or α-hydroxy groups at the 2-, 4- or 6-position. Thus, it is surprisingly unnecessary to protect the hydroxyl groups at the non-5-position of the starting actinamine with much difficulty and labor before carrying out the process of the invention. A further advantage of the process according to the invention is that at the 3'-position, either alone or in masked or latent form, a carbonyl group can be selectively formed in an elimination step. The carbonyl group at the 3'-position is a particularly important but difficult to obtain feature of the compounds of formula (I) obtainable by the process of the invention.

Scheme B illustrates the novel preparation process of the present invention. In Scheme B, R, R ₂ and R ₃ are as defined for formula (I), while T is aralkoxycarbonyl or optionally halogenated alkoxycarbonyl, preferably benzyloxycarbonyl.

An even closer example of carrying out the process of the present invention is illustrated in Scheme C. Although both α and β anomer can be prepared according to step (1) of this scheme, it is preferred to prepare the β anomer by separating it from the α anomer in any subsequent step of the process. In the initial coupling reaction, a commercially available enantiomeric sugar can be used when the β-anomer will be formed in large amounts. However, any intermediate or end product may be epimerized to increase the amount of the β-anomer. It is also noted that anomeric mixtures can also be used as active ingredients of antimicrobial agents, since they will exert biological activity with respect to the active anomer contained therein.

Although the compounds of formula (I) having the β-configuration are undoubtedly biologically active anomers, the term is not to be construed in a restrictive manner, since the a-anomers may be present without impairing the action of the β-anomer. However, the aanome of a spectinomycin analog can be anomerized to the corresponding β-anomer. The anomers may be separated from each other after any step of the process of the invention.

In carrying out the process of the present invention, new intermediates are formed. These are compounds of the formula II, wherein the substituents have the meanings given above, and derivatives of the compounds of the formula II, in which the 2'-hydroxyl group can be acylated, preferably acetylated.

Furthermore, the novel compounds of formula (III), ame10 of the formula wherein the substituents are an alkyl group having the previously defined and R is C1-4 alkyl _down.

In the case of the compounds of formula (Ib1b), the hemiketal may be in partially or fully open ketone form. These two forms may be in equilibrium with each other as shown in Scheme D.

Turning now to the detailed description of the process according to the invention, it should first be pointed out that the process can actually produce spectinomycin itself. Spectinomycin of Formula VIII is an aminocyclitol antibiotic that is unique in structure to a single sugar component fused to an actinin by both a β-glucoside linkage and a hemiketal linkage. The process of the present invention for the preparation of these uniquely fused spectino25 micin analogs is in fact a synthesis involving a sugar derivative and a protected actinin. The sugar may be of natural or artificial origin, or may be chiral or achiral.

The process according to the invention may comprise two essential steps. Possible variants for carrying out these steps are shown in Schemes E and F.

The la. Before proceeding step 2, the two amino groups of an actinamine derivative of formula Vla must be protected by the application of known protecting groups such as aralkoxycarbonyl, aryloxycarbonyl or optionally halogenated alkoxycarbonyl groups. For example, the preparation of benzyloxycarbonyl and tert-butoxycarbonyl derivatives of amino acids and the deprotection of protecting groups are described in detail by Boissonas, R. A., in "Synthesis of Peptides from Selectively Removable Amino Protective Groups". in Advances in Organic Chemistry, 3, 159-190 (1963). The use of the tert-butoxycarbonyl 45 protecting group for the amino group is further described in the September 1976 publication "BOC-OH" ALDRICH Technical Infonnation Bulletin. The use of the trichloroethoxycarbonyl group to protect the amino group is described by Windholz et al., Tetrahedron Letters, 2555 50 (1967). '

The actinins used as starting materials in the process of the invention are known in the art, such as those described by Suami et al., Bull. Chem. Soc. Jap., 43, 1843 (1970). Similarly, sugars used as starting materials in the process of the invention are commercially available or can be obtained by methods known per se, such as those described by Mochalin et al., Chem. Comp., 699 (1977) (Kim. Heterocyclosojegyiny., 867 (1977)) can be prepared as described.

1a. Step (a) is carried out by coupling an actinamine of formula (VIIa) with a sugar of formula (VIIa), (VHb) or VIIc). This coupling reaction in 65 Ν, Ν-dimethylformamide or similar solvent,

-2181535, for example, in diethyl ether, tetrahydrofuran or dimethoxyethane, sometimes in the presence of a base. The reaction can be carried out very effectively under a nitrogen atmosphere at normal temperature and pressure, as in the case of a similar reaction by Lemieux et al. J. Chem., 51, 53 (1973). Thus, the reaction is generally carried out at 0 ° C and 45 ° C, using 0.01-0.5 molar solution of activated sugar, 0.01-0.5 molar solution of actin derivative, and 0 molar actinine per mole. Using 2-4 to 10 moles of sugar. Preferably, the reaction is carried out at 20-30 ° C in dimethylformamide, with the sugar: actinamine ratio being selected between 3: 2 and 2: 3. The reaction time may vary from 4 hours to 1 week, but preferably from 24 to 48 hours.

The resulting compound of formula (Va), (Vb) or (Ve) is generally isolated from the reaction mixture by concentration or concentration and vigorous mixing with excess water. The resulting precipitate was then taken up in chloroform and then evaporated to dryness to give the crude oxime intermediate. The α and β anomers can then be separated by chromatography on a silica gel column using methanol: chloroform (1:99 to 2:98) as eluent. Of course, other isolation methods such as extraction, crystallization, chromatography or any combination thereof may be used.

The lb. In step (b), the oxime group of a compound of formula Va is removed to form a cyclic hemiketal of formula IVa. This operation is described by Le mieux et al., Can. J. Chem., 51, 19 (1973) or by deoxylation methods similar to those described by Mallams et al., J. Chem. Soc. Perkins 1,1097 (1976). For example, the crude oxime or an already isolated α- or β-anomeric oxime may be dissolved in a solvent and then acetaldehyde and hydrochloric acid added. The initial concentration of oxime in the solvent is adjusted to 0.01 to 1.5 molar, preferably 0.1 to 0.3 molar, while the molar ratio of acetaldehyde to oxime is adjusted to a ratio of 1: 1 to 80: 1. At the same time, 1N hydrochloric acid was added in a molar ratio of about 2: 3 to oxime. The reaction mixture is stirred at room temperature for 3.75 hours to 5 days, then optionally sodium bicarbonate is added and stirring is continued for another 15 minutes. Alternatively, after prolonged stirring, the reaction mixture can be directly subjected to chromatography, whereby the silica gel directly removes the hydrochloric acid during the purification step. 50

A compound of formula (IVa) may be isolated by one of the conventional isolation methods mentioned above. One such method is to filter the reaction mixture and then evaporate the filtrate in vacuo to give the crude product. Here again, the crude product is subjected to chromatography on a column of silica gel 55, eluting with a 95: 5 mixture of chloroform and methanol. TLC analysis revealed that the α- and β-anomers of general formula IVa (60) could be separated by pooling the fractions containing the same substance and then evaporating in vacuo.

The lb. Bases other than sodium hydrogencarbonate may be used in the deoxylation step of Step 1, but it is essential to ensure moisture retention in order to prevent degradation of the ring intermediates which are essential for the final products of the present invention. Therefore, we need to use bases that do not attack the acetates, such as hydroxides in excess. Useful bases include, for example, benzoates or potassium hydrogen phosphate.

Examples of suitable solvents for the reaction include acetonitrile, tetrahydrofuran, diethyl ether or dimethylformamide. The preferred solvent is acetonitrile.

That's down. In step (b), one or two substituents are simultaneously removed from the sugar moiety of the compound of formula (TVa) and the carbonyl group in the 3'-position is eliminated by β-elimination from the 4'-position to produce a compound of the formula (IIIa). Further elimination may result in the formation of a 4 ', 5' double bond. That's down. Step (a) is carried out at 0-80 ° C for 2 hours to 1 week in the presence of a base system. Suitable bases for this purpose include, for example, potassium bicarbonate, triethylamine, pyridine or alcoholates as bases. The use of a base system consisting of potassium bicarbonate and acetonitrile is preferred. 0 to 20 molar equivalents of base may be used, but 1 to 10 molar equivalents are preferred.

That's down. However, the manner in which steps 1 to 4 are carried out depends on the quality of the protecting groups on the sugar moiety and the actinamine moiety. Generally, the sugar moiety is easier to remove than the actinamine moiety. That's down. In any event, step (b) should be considered as a novel process for the preparation of the important 3'-carbonyl group (or its latent form) under mild conditions and selectively by elimination.

In our experiments with 4'- and 6'-derivatives having elimination, it has been found that, for example, acetic acid in acetates is benzoic acid in benzoates, benzyl alcohol in benzyl esters and hydrogen halides in the elimination. However, they are down. are only examples.

The intermediates used in the process of the invention, but in particular the intermediates. The intermediates obtainable in Step (b) can be used to prepare a wide variety of analogs. These methods of preparation are characterized by the functional group change of the starting intermediate in known methods, such as halogenation, reduction, oxidation, or chain extension reactions.

In carrying out the process of the present invention, in some cases, it is described below. As a consequence of the elimination step, the oxygen substituent attached to the 3'-carbon atom may migrate to the 2'-oxygen and at the same time form a 3'-carbonyl group. This phenomenon occurs when a compound of formula (IVa) is converted to a compound of formula (IIIa) or a compound of formula (TVc) to a compound of formula (lile). In other cases, however, the 3'-oxygen substituent does not migrate to form a masked or latent 3'-carbonyl group, these are enol derivatives. For example, the conversion of a compound of formula IVb to a compound of formula IIIb is mentioned. Enol derivatives may exist in the form of hemiketals or open ketone isomers or as mixtures of the two.

However, both types of preparation are novel, useful and selective, which ultimately yields a compound analogous to spectinomycin and bearing a 3'-carbonyl group. Unique chemical properties of masked or latent 3'-carbonyl derivatives

-3181535 which allow the chemical modification of these compounds by known methods such as halogenation, alkylation, acylation or oxidation. Finally, the masked or latent 3'-carbonyl group is much more stable, especially against bases.

An intermediate of formula (IIIa) may also be isolated from the reaction mixture by conventional means such as precipitation, crystallization or concentration followed by chromatography.

See ld. In step 1, the protecting groups at one or more positions of the sugar ring are removed. Usually, a protecting group is present at the 2 ', 3' or 6 'position and, depending on its nature, an acid and / or a base may be used. When a base is used to convert a compound of formula (IIIa) into a compound of formula (IIa) or a compound of formula (IIIa) into a compound of formula (IIc), hydrolysis at a temperature between -10 ° C and + 50 ° C hours. Preferably, the reaction is carried out at 20-30 ° C for a reaction time of 1-20 hours.

Useful alcohols include methanol, ethanol and isopropanol, although the use of methanol is preferred. Any base which does not degrade the product to be produced may be used. For example, sodium bicarbonate, potassium bicarbonate, pyridine, potassium hydrogen phosphate, triethylamine or potassium sodium tartrate may be used, but potassium hydrogen phosphate is preferred. When converting a compound of formula (IIb ') to a compound of formula (IIb), the 6'-acetyl group may first be selectively removed using one of the abovementioned alcohols and bases (in which case the 3'-methoxy group will remain chemically intact).

Acid hydrolysis may also be used to remove protecting groups on the sugar ring. For example, after removal of the 6'-acetyl group from a compound of formula IIIb with a base as described in the preceding paragraph, the 3'-protecting group may be removed by acid treatment to give a compound of formula IIb. Alternatively, a compound of formula (IIb ') can be converted into a compound of formula (IIb) in a single step by acid catalysis.

The deprotection by acidic treatment is carried out at a reaction temperature of 0 to 80 ° C, preferably 20 to 30 ° C, for 1 hour to 3 days, preferably 2 hours to 2 days. Suitable acids for this purpose are, for example, hydrochloric acid, p-toluenesulfonic acid or phosphoric acid; hydrochloric acid is preferred. Treatment solvents include water and tetrahydrofuran, or a mixture of water and dimethoxyethane, and methanol or ethanol. Methanol or a mixture of water and tetrahydrofuran is preferred.

In some cases, it may be advantageous to refer to ld. combining step 2 with step 2 by using a reaction medium of the same type as described in ld. fulfills the requirements for deprotection as described in step. For example, if Step 2 is carried out in isopropanol in the presence of pyridine, the intermediate (IIIa) is formed into spectinomycin. That is, in addition to the deprotection (and saturation of the 4'- and 5'-positions) of nitrogen under the influence of palladium catalyst and hydrogen, the 2'-position acetyl is also deprotected as a result of the alcoholic reaction with the basic solvent system.

In some cases, it may be advantageous to refer to ld. skipping step or part of this step, whereby cleavage of the protecting group occurs in a biological system in which the active end product is to be utilized.

The reaction parameters to be followed in step 2 for the deprotection of the actinamine moiety depend on the particular groups, i.e., the nature of the T groups. By appropriately selecting the aforementioned groups and the reaction parameters provided for the deprotection, a 4 ', 5' double bond can be left intact or reduced during the deprotection. For example, when the protecting group is a benzyloxycarbonyl or aralkoxycarbonyl group, the cleavage may be carried out at a hydrogen gas pressure of 200 to 14 atoms in the presence of conventional catalysts such as palladium on carbon, palladium on carbon, palladium on barium sulfate. Said catalysts may be used in suspension in solvents such as isopropanol, anhydrous ethanol, ethyl acetate, toluene or tetrahydrofuran.

Alternatively, compounds having an alkoxycarbonyl or aryloxycarbonyl group in place of the T substituents may be deprotected with an acid in a suitable solvent such as nitromethane or methylene chloride.

If it is a T-alkoxycarbonyl group, the cleavage of these groups is preferably carried out in the presence of zinc.

In addition, any isolation methods well known in the art may be used to isolate any compound of formula (I). One such method of isolation is by evaporating the excess solvent and preparing a crystalline salt of the compound to be isolated. For the preparation of these salts, for example, solutions of acids such as p-toluenesulfonic acid, hydrochloric acid, hydrobromic acid or hydrogen iodide may be used, and water, methanol, ethanol, isopropanol, diethyl ether, 1,2 -dimethoxyethane or p-dioxane. The salt is isolated by filtration and direct crystallization or by evaporation of the solvent followed by recrystallization from a suitable solvent.

Alternatively, the crude compounds of Formula I may be purified by column adsorption from a weakly acidic ion exchange resin such as Amberlite IRC-50 or CG-50, followed by the use of a solvent such as hydrochloric acid, hydrobromic acid, hydrogen iodide or sulfuric acid. containing water, methanol, ethanol, diethyl ether, tetrahydrofuran, 1,2-dimethoxyethane or p-dioxane.

Similarly, salts of the compounds of formula (I) formed on the free hydroxy groups can be converted to the corresponding free analogs by passing a solution of the salt in, for example, water, methanol, ethanol, tetrahydrofuran or 1,2-dimethoxyethane, a basic ion exchange resin, e.g. hydroxyl form Dowex 1-X8 resin and evaporate the eluate.

Each of the foregoing operations may be performed on a mixture of different anomers or on the desired β-anomer itself, which may be isolated at any stage of the process of the invention. The remaining steps can now be performed with the β-anomer to give the biologically active end product.

Preferably, the β-anomer is separated from the reaction mixture obtained after step 1, i.e. the coupling of the sugar and actinine moieties, and the step 2 is performed only on the β-anomer, yielding only biologically active end products.

-4181535

The individual anomers may be separated from the anomeric mixtures by methods well known to those skilled in the art by conventional modifications of resolution. For example, the compound of formula (IV) can be separated by applying a column of silica gel to elute the desired β-anomer and eluting the column with methanol: chloroform 1:99 to 2:98. Alternatively, the corresponding β-anomer can be isolated from the compound of formula (V) in the same manner.

In a further preferred embodiment of the process of the invention, an enantiomeric sugar is used in step 1, i.e., the coupling step, to obtain a relatively high β-anomeric concentration, which is favorable for the formation of the β-structure. For example, D-arabinose results in the formation of β- and α-oximes in a ratio of approximately 4: 1. Thus, in order to avoid the need for laborious and time-consuming separation operations to obtain the β-configurated intermediates or products, these enantiomers may be commercially available or not costly to obtain. The use of such an enantiomer results in an intermediate or end product which is rich in the desired β-anomer and can thus be converted to a substantially pure β-anomer by one of the methods described above or used without further purification.

The acid addition salts of the compounds of formula (I) may also be prepared, for example, by neutralizing a compound of formula (I) with a suitable acid to a pH of less than 7.0, preferably between 2 and 6. Suitable acids for this purpose include hydrochloric acid, sulfuric acid, phosphoric acid, sulfamic acid or hydrobromic acid. It should be noted that the salts of the compounds of the formula I with acids and bases can be used for the same biological purposes as the compounds of the formula I themselves.

The compounds of the formula I inhibit the growth of microorganisms in various environments. For example, the compounds of formula (I) are active against Escherichia coli and can thus be used against this microorganism in the sludge formed during papermaking. The β-anomers of formula (I) can also be used to extend the lifespan of cultures of Trichomonas fetus, Trichomonas hominis and Trichomonas vaginalis by depriving these cultures of Escherichia coli contamination. The β-anomers are also effective against Bacillus subtilis and can thus be used to reduce or prevent the odor caused by this microorganism in fish or fish crates. The same anomers can be used in a mycology laboratory to wash laboratory racks and equipment. Panomers are also effective against Klebsiella pneumoniae.

The compounds of the formula I are effective in the treatment of bacterial infections, such as gonorrhea, as well as tumors in mammals, including man.

The compounds of the present invention may be converted into pharmaceutical compositions in conventional manner. These formulations, usually in unit dosage form for human and veterinary use, can be, for example, tablets, capsules, pills, powders, granules, sterile parenteral solutions or suspensions, eye drops, oral solutions or suspensions, and water-in-oil emulsions. containing an effective amount of a compound of formula I.

For oral administration, solid or liquid dosage units may be prepared. For the preparation of solid preparations such as tablets, a compound of formula (I) is usually formulated with conventional carriers and / or diluents or other excipients such as alkali, magnesium stearate, dicalcium phosphate, magnesium aluminum silicate, calcium sulfate, starch, , gum arabic or methyl cellulose. Capsules may be prepared by mixing the active ingredient with an inert drug diluent and filling the resulting mixture in a hard gelatin capsule of appropriate size. Soft gelatine capsules are prepared by encapsulating a suspension of the active ingredient in a pharmaceutically acceptable vegetable oil, light petroleum oil or other inert oil in a suitable device.

Liquid dosage units for oral administration such as syrups, elixirs, or suspensions may also be prepared. The water-soluble active compounds may be dissolved in an aqueous carrier together with sugar, aromatic flavors and preservatives to form a syrup. Elixir may be prepared using an alcoholic carrier such as ethanol, and a suitable sweetener such as sugar or saccharin and an aromatic flavoring.

Suspensions may be formulated with an aqueous carrier using a suspending agent, for example, gum arabic, gum tragacanth or methylcellulose.

For parenteral administration, liquid dosage units are prepared using the active ingredient and a sterile vehicle, preferably water. Depending on the vehicle or concentration, the active ingredient may be dissolved or suspended in the vehicle. In preparing solutions, the active ingredient is dissolved in water for injection and then sterilized by filtration before filling into a suitable ampoule or vial. Advantageously, excipients such as topical anesthetics, preservatives and buffering agents may be dissolved in the vehicle. To increase stability, the composition may be frozen after filling into the vial and the water removed under vacuum. The dry lyophilized powder is then sealed in the vial and a ready-to-use vial containing water for injection is provided to reconstitute the composition prior to use. Parenteral suspensions can be prepared in substantially the same manner except that the active ingredient is suspended in the vehicle instead of being dissolved and sterilization is not accomplished by filtration. The active ingredient may be sterilized by treatment with ethylene oxide prior to suspension in the sterile vehicle. Preferably, a surfactant or wetting agent is added to the composition to ensure a uniform distribution of the active ingredient. _x

The active ingredient may also be delivered in the form of a rectal suppository. This dosage form is particularly important if the mammal cannot be easily treated with another dosage form, for example, orally or by insufflation, e.g., in infants or persons with debility. The active ingredient may be incorporated into any known suppository base by any of the methods well known in the art. Examples of such materials include cocoa butter, polyethylene glycols (e.g., Carbowax brand names), polyethylene sorbitan monostearate, and mixtures thereof with other compatible materials to modify the melting point or dissolution rate. The weight of these rectal suppositories generally ranges from 1 g to 2.5 g.

The term "dosage unit" refers to physically discrete units suitable for unit dose administration.

-5181535 for human or animal administration, each containing a predetermined amount of the active ingredient to achieve the desired therapeutic effect, together with the necessary pharmaceutical carrier, diluent and / or other excipients. The characteristics of the pharmaceutical compositions of the present invention are determined by the action of the compounds of formula (I) and the known limitations of the preparation of the active ingredient. Dosage units for use in the present invention include, for example, tablets, capsules, pills, suppositories, powder packets, wafers, granules, teaspoonfuls, drops, ampoules, vials, metered dose aerosols and the like.

Naturally, an effective amount of the active ingredient is used in medical treatment. It will be appreciated by those skilled in the art that the effective amount, i.e., dose, will depend on a variety of factors. These factors include, for example, the route of administration and the potency of the compound. For the treatment of tumors and bacterial infections, single doses of about 2 to about 4,000 mg may be administered parenterally to man. More often, a single dose is between 5 mg and 200 mg. Single oral and rectal dosages range from 5 mg to 5000 mg, more typically from 10 mg to 2500 mg.

The invention is further illustrated by the following Reference Examples and Embodiments. It will be apparent to those skilled in the art that the appropriate stereoisomers may be used instead of the specific compounds mentioned in the examples.

Reference Example 1

Preparation of 5-O- (3 ', 4'-di-O-Acetyl-2'-deoxy-2'-oxyimino-D-arabinopyranal) -N, N'-bis (benaloxycarbonyl) -actinamine (Reaction G )

To a solution of 7.2 g (27.1 mmol) of 3,4-di-O-acetyl-2-deoxy-2-nitroso-N-D-arabinopyranoal chloride in 75 mL of dimethylformamide was added 11.86 g (25 mL) of (0 mmol) of Ν, Ν'-bis (benzyloxycarbonyl) -actinamine in 50 ml of dimethylformamide and the resulting mixture was stirred under nitrogen at room temperature for 19.5 hours. The reaction mixture was then poured into 1 L of water with vigorous stirring. The aqueous phase is then decanted and the crude portion is taken up in 300 ml of chloroform. A small amount of water was collected and the chloroform solution was evaporated in vacuo to give 12.6 g of a brittle white foam. The whole aqueous phase was then extracted with chloroform (3 x 100 mL), and the combined extracts were washed with water (50 mL), brine (50 mL) and dried over anhydrous sodium sulfate. The solution was concentrated to 40 mL and the concentrate was added to 300 mL of water with vigorous stirring. The slight chloroform layer was separated and the solvent was evaporated in vacuo to give 6.18 g of a white foam which was added to the first foam. The proton magnetic resonance spectroscopy (PMR) analysis of the crude product thus obtained showed that most of the dimethylformamide was removed in this way. Purification by chromatography on 1.25 kg silica gel (gradient elution with methanol / chloroform) first gave 1.33 g (1.89 mmol, 7.6%) of the pure α-anome of the title compound and 2.86 g of pure β-anometry of the title compound and 3.17 g of additional β-anomer are obtained which are approximately 90% pure.

Identification of the α-anomer:

IR (CHC1 ₃₎ 3550, 3400, 3070, 2980, 1745, 1686, 1675 (shoulder), 1484, 1449, 1364, 1333, 1235, 1167, 1026 and 669 cm in ^{the fourth}

PMR (CDCl₃): δ 7.40 (s, 10, aromatic), 6.07 (s, 1 H), 5.95 (d, J = 3 Hz, 1 H, _a), 5.20 ( m, 7), 3.3-4.6 (m, 10), 3.12 and 3.08 (s's, 6, NCH ₃ ), 2.12 and 2.03 (s's, 6, CH ₃ ).

Mass spectrum (for the tetra-trimethylsilyl derivative (hereinafter referred to as the tetra-TMS derivative)): 991 (M ⁺ ), 689 and 673.

Carbon Magnetic Resonance Spectroscopy (hereinafter referred to as CMR; D ₆ -aceton):? 170.1, 169.8, 157.8, 156.5, 150.1,

138.1, 129.1, 128.8, 128.4, 91.7, 86.9, 79.1, 174.4, 69.1,

68.6, 68.3, 67.8, 67.3, 60.3, 57.7, 31.5, 31.4, 20.8 and 20.5.

130-140 ° C (dec.).

Molecular weight of the tetra-TMS derivative on C4 ₅ H ₇₃ N ₃ O ₄ S ₄ S _{4 by} high resolution mass spectrometry:

calcd: 991.4169, found: 991.4190.

Identification of the β-anomer:

IR (CHC1 _3): 3500, 3100, 2970, 1748, 1686, 1672 (shoulder) cm 1486,1449,1370,1337,1235,1167,1107,1024és700.

PMR (CDCl 3): δ 7.40 (s, 10, aromatic), 6.40 (s, 1H), 6.05 (d, J = 3 Hz, 1, H _{3 (} ), 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, CH ₃ ).

Mass spectrum (for the tetra-TMS derivative): 991 (M ⁺ ), 990, 976, 975, 689 and 673.

CMR (D ₆ -aceton): δ 170,7,169,9,157,7,157,1,149,1,137,9,

129.1, 128.7, 128.3,92.3, 85.3, 74.7, 70.8,69.4, 67.3, 62.0,

60.2, 31.4, 20.8 and 20.5.

Melting point: 140-155 ° C (dec.).

Molecular weight measured on tetra-TMS derivative by high resolution mass spectra for C ₄₅ H ₇₃ NaO ₁₄ Si 4:

· calculated. 991.4169, found: 991.4229.

la. PREPARATION

Preparation of 5-O- (3 ', 4', 6'-tri-O-acetyl-2'-oxyimino-2'-deoxy-αL-glucopyranosyl) -N, N, bis (benzyloxycarbonyl) -actinamine ( Scheme H).

To a solution of 7.09 g (21 mmol) of 3,4,6-tri-O-acetyl-2-nitroso-2-deoxy-L-glucopyranosyl chloride in 150 mL of dimethylformamide, 14.22 g (30 mmol) of N, N'-bis (benzyloxycarbonyl) -actinamine was added and the reaction mixture was stirred for 19 hours, at which point the pyranosyl chloride was no longer detectable. The resulting solution was then concentrated at 45 ° C and the residue was stirred at room temperature for 1 hour. Dissolve in 5% methanol in chloroform, apply to a column of 3 kg wet silica gel, elute with said solvent, and use approximately 10 liters of eluent for continuous thin layer chromatography (5% methanol as eluant). The fractions containing pure product were combined and evaporated to give the title compound (7.86 g, 48%).

Circular Dichroism (hereinafter referred to as CD; recorded in methanol) (θ) ^ "ιημ = + 18 900 ± 1300 Specific Rotation [a] g = - 53 '(6.0.9%, acetone)

PMR (CDCl 3): 2.02 (s, 9H), 3.03 (s, 6H), 5.04 (s, 4H), 6.20 (s, 1H) and 7.32 (s, 10H).

CMR (D ₆ -aceton): δ 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 and 20.6 p. p. m.

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

lb. PREPARATION

5-O- (4 ', 6'-di-O-acetyl-3'-O-methyl-2'-deoxy-2'-oximino-PD-glucopyranosyl) -N, N'-bis (benzyloxycarbonyl ) -actinamine (Scheme I)

To a solution of 60.8 g (196 mmol) of 4,6-di-O-acetyl-3-O-methyl-2-nitroso-αD-glucopyranosyl chloride in 1.1 L dimethylformamide was 97.7 g (201 mmol). N, N'-Bis (benzyloxycarbonyl) -actinamine was added and the reaction mixture was stirred under nitrogen at room temperature for 45 hours and then concentrated to a thick syrup at 30 ° C under high vacuum. This is then added in small portions to 3 liters of ice-cooled water with vigorous stirring. The white precipitate formed is filtered off, redissolved in 3 L of chloroform, the chloroform phase is separated from the residual water and dried over anhydrous sodium sulfate and finally evaporated. This gives 143 g of a viscous white foam which, according to nuclear magnetic resonance spectroscopy (NMR), contains about 7% dimethylformamide.

Gradient elution of 3.5 kg silica gel with methanol and chloroform allows the products to be roughly separated. The fraction containing the β-anomer was crystallized from acetone to give 4.4 g of pure β-anomer. A further 1.5 g of β-anomer can be isolated by chromatography on the mother liquor.

IR: 3500, 3310, 2920, 1750, 1690, 1459, 1240, 1175, 1105, 1403, 735 and 711 cm in ^{the fourth}

PMR (CDCl 3): δ 7.32 (s), 5.41 (s), 5.11 (s), 3.5-4.5 (m), 3.33 (s), 3.04 (s) ) and 2.02 (s).

CMR (CDCl 3): δ 170.6, 169.6, 157.7, 156.5, 150.1, 136.5,

128.5.127.8.95.7.92.7, 77.1, 73.2, 70.1,67.54, 65.0, 56.8,

29.7, 20.8 and 20.5 p. p. m.

Mass spectrum for the tetra-TMS derivative: 1035 (M ⁺ ). Melting point: 214-216 ° C

Reference Example 2

Preparation of N, N'-Bis (Benzyloxycarbonyl) -5'-demethyl-3'-O-acetyl-4 '- (R) -acetoxy-3' - (R) -dihydro-spectinomycin (Scheme J) ml 2.32 g (3.3 mmol) of 5-O- (3 ', 4'-di-O-acetyl-2'-deoxy-2'-oxyimino-D-arabinopyranosyl) -N, N' are dissolved in methylnitrile. bis (benzyloxycarbonyl) -actinamine followed by 5.0 ml (89.4 mmol) of acetaldehyde and

1N HCl (2.5 mL, 2.5 mmol) was added. After stirring at room temperature for 3.75 hours, sodium sulfate was added and stirring was continued for 15 minutes. The reaction mixture was filtered and the filtrate was concentrated in vacuo to give a white precipitate (26 g). The latter was chromatographed on 200 g of silica gel with methanol-chloroform to give 1.45 g (2.11 mmol, 64%) of the title compound as a white solid, m.p. 135.0-12.9 ° C.

IR (CHC1 _3): 3450, 3050, 1748, 1684, 1881, 1447, 1362, 1333, 1238,1160,1050, 1020 and 680 cm in ^{the fourth}

PMR (CDCl 3): δ 7.4 (s, 10, aromatic), 3.4-5.4 (m), 3.05 (s, 6, NCH ₃ ), 2.10 (s, 3, COCH 3). and 1.95 (s, 3, COCH ₃ ).

CMR (D _e -aceton): δ 170.2,169,8,157,5,156,9, 138,0,129.1, 128.3, 94.6, 91.5, 74.7, 72.4, 67.3, 67 , 0, 66.3, 65.5, 61.7,

61.2, 57.5, 31.6 and 20.6.

Mass spectrum for the tetra-TMS derivative: 904 (M ⁺ ).

2a. reference example (Scheme K)

7.0 g (9.03 mmol) of α-5-O- (3 ', 4', 6'-tri-O-acetyl-2'-deoxy-2-carbonyl-ol-glucopyranosyl) -N, A solution of N'-bis (benzyloxycarbonyl) -actinamine, 7 ml of acetonitrile, 14.2 ml of acetaldehyde and 17.0 ml of 1 N hydrochloric acid was stirred at room temperature for 5 hours, followed by addition of 60 g of anhydrous sodium sulfate and stirring. Continue for 10 minutes. The precipitated solid was filtered off and washed with acetonitrile. The filtrate was evaporated and the residue was taken up in 15 ml of a 1: 1 mixture of chloroform and ethyl acetate and the resulting solution was chromatographed on 150 g of silica gel using the same solvent mixture. The eluate was evaluated by TLC using 5% methanol in chloroform as the eluent and the fractions containing pure product were pooled and evaporated to give 5.96 g (87%) of the corresponding hemiketal triacetate.

Specific rotation [α] = -52 '(0.7%, chloroform). PMR (CDCl 3): δ 2.06 (9H, s), 2.88 (3H, s), 3.08 (3H, s), 4.90 (s), 5.13 (4H, s) and 7 , 38 (10, s).

CMR (d ₆ -aceton): δ 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 and 20.0 p. p. m.

2b. Reference Example

Preparation of N, N'-bis (Benzyloxycarbonyl) -3'-O-methyl-4 '- (R) -6'-diacetoxy-3' - (S) -dihydro-spectinomycin (Scheme L).

300 mg (0.4 mmol) of 5-O- (4 ', 6'-di-O-acetyl-3'-O-methyl-2'-deoxy-2'-oximimo-p-D-glucopyranosyl) -N, N To a suspension of 1'-bis (benzyloxycarbonyl) -actinamine in 12 ml of acetonitrile was added 1.2 ml of acetaldehyde and 0.18 ml of 1N hydrochloric acid, and the resulting mixture was stirred for 60 hours. The resulting homogeneous solution is then stirred with anhydrous sodium sulfate for 20 minutes and then filtered. Chromatography of the precipitated product on silica gel (10 g) with ethyl acetate (15% chloroform) as the eluent afforded the title compound (150 mg). PMR (CDCl 3): δ 7.32 (s), 5.1 (s), 4.68 (s), 3.5 (s), 3.1 and 2.1. CMR (I) ₆ -aceton): δ 171,0,170,3,157,8,138,2,129,2,128,5,

95,8,93,8,84,5,75,2,73,1,69,8,67,4,66,4,65,2,63,7,61,2,

57.9, 57.5, 30.8, 29.8, 21.0 and 20.7 p. p. m.

Mass spectrum for tri-TMS [m / e]: 948 (M ⁺ )

933 (M-15).

-7181535

2c. PREPARATION

Preparation of N, N'-bis (Benzyloxycarbonyl) -6'-acetoxy-spectinomycin-3'-methylenol ether (Scheme M)

N, N'-Bis (benzyloxycarbonyl) -3'-O-methyl-4 '- (R) -6'-diacetoxy-3' - (S) -dihydro-spectinomycin (101.3 mg, 0.136 mmol)

A solution of 5.5 ml of acetonitrile was stirred with 350 mg of potassium bicarbonate at room temperature for 6 days, then the reaction mixture was filtered through Celite and the filtered bicarbonate washed with additional acetonitrile. Concentration of the filtrate gave 80.5 mg of the title compound.

PMR _(D -acetone): δ 7.4 (s), 5.1 (s), 4.8 (s), 4.75 (s), from 4.6 to 4 (m), 3.9 ( s), 3.5 (s), and 2.0 (s).

CMR (D ₀ -aceton): δ 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, 61.08,

60.7, 57.8, 55.6, 30.8 and 20.75 p. p. m.

Mass spectrum for tri-TMS derivative [m / e]: 888 (M +) and 873 (M-15).

Reference Example 3

Preparation of N, N'-Bis (Benzyloxycarbonyl) -5'-demethyl-spectmomycin (Scheme N) Dissolve 1.32 g (1.92 mmol) of N, N'-bis-benzyloxycarbonyl in acetonitrile. 5-Demethyl-3'-O-acetyl-4'-R-acetoxy-3 '- (R) -dihydro-spectinoinycin and 1.0 g (7.24 mmol) of anhydrous potassium carbonate were added to the resulting solution. After stirring at room temperature for 20 hours, the reaction mixture was filtered and the precipitate was washed with additional acetonitrile (20 mL). The solvent was removed in vacuo and the precipitate was dissolved in methanol (50 mL) and potassium hydrogen phosphate (1.4 g, 8.04 mmol) was added to the resulting solution. The resulting mixture was stirred at room temperature for 22 hours, then filtered and the solid residue thoroughly washed with methanol. Removal of the solvent in vacuo gave a resinous orange precipitate. Rapid filtration over 40 g of silica gel with ethyl acetate gave 950 mg of a white precipitate. Purification by preparative thin layer chromatography on 2000 μ silica gel plates with ethyl acetate as the eluent afforded the pure title compound as a cl.

IR (CHC1 _3): 3600, 3100, 1748, 1695, 1443, 1333, and 700 cm 1156.1111 ^4th

PMR (CDCl ₃ ): δ 7.40 (s, 10, aromatic), 5.13 (s, 4), 3.1-5.0 (m), 3.05 and 2.95 (s's, 6, NCH ₃ ).

CMR (D _e -aceton): 8201,5,157,1,156,8,137,5,128,5,128,2,

127.8, 97.7, 92.4, 74.4, 74.1, 66.6, 65.8, 65.0, 60.8, 56.7,

38.2 and 30.8.

3a. PREPARATION

Preparation of N, N-bis (benzyloxycarbonyl) -6'-hydroxy spectinomycin (Scheme O) mg (0.04 mmol) of N, N-bis (benzyloxycarbonyl) -6'-acetoxy spectinomycin-3 To a solution of 1-methylenol ether in 0.65 ml of tetrahydrofuran was added 0.5 ml of a 4N aqueous hydrochloric acid solution, and the resulting mixture was stirred at room temperature for 28 hours. The reaction mixture was diluted with chloroform (25 mL) and washed with saturated sodium bicarbonate solution (4 mL) and water (4 mL). The organic phase is then separated, dried over anhydrous sodium sulfate, filtered and evaporated to give the title compound (22 mg).

PMR (CDCl ₃ ): δ 7.25 (s), 5.1 (s), 4.75 (s), 4.5-3.5 (m) and 3.0 (s).

CMR (CDCl ₃ ): δ 157.5,156,5,136,3,128,5,127,8,96,8,91,0, 77,0,74,3,74,0,72,1,67,6,67,5 , 65.8,65,2,64,4,61,7,57.1, 39.0 and 29.7 ppm

3b. PREPARATION

Preparation of N, N-bis (benzyloxycarbonyl) -6'-hydroxy spectinnoticin (Scheme P) mg (0.143 mmol) of N, N'-bis (benzyloxycarbonyl) -6'-hydroxy spectinomycin-3 ' To a solution of methylenol ether in 2.2 ml of tetrahydrofuran was added 1.6 ml of a 4N aqueous hydrochloric acid solution, and the resulting mixture was stirred at room temperature for 24 hours and then treated with 3a. as described in reference example. The title compound thus obtained exhibits the same behavior and properties as TLC. Reference Example. The two products were combined and purified by chromatography on silica gel, eluting with ethyl acetate: chloroform 1: 1. PREPARATION

Preparation of N, N'-Bis (Benzyloxycarbonyl) -2'-O-acetyl4 ', 5'-didehydro-spectinomycin (Scheme Q)

In a flask fitted with a distillation head, a mixture of 14.58 g (146 milliequivalents) of anhydrous potassium bicarbonate and 384 ml of acetonitrile is heated until 40 ml of solvent are distilled off. The remaining slurry was then cooled to room temperature under a nitrogen atmosphere. the hemiketal triacetate prepared according to the referendum example is added. The resulting reaction mixture was then stirred at room temperature for 41 hours. Thin layer chromatography on 5% methanol in chloroform as eluent indicated that the starting material had been completely converted. The precipitate was filtered off, washed and the filtrate was combined with the washing liquid. The resulting mixture was evaporated to give 14.51 g of a foam. This was then dissolved in chloroform and the resulting solution was cooled to give a precipitate (7.55 g). This was recrystallized from 20 ml of chloroform to give 3.42 g of the title compound, m.p. 149-152 ° C. Chromatography of the combined mother liquor on 800 g of silica gel with a 1: 4 mixture of methanol and chloroform as the eluent afforded an additional 3.37 g of product (55% overall yield).

UV spectrum (C ₂ H ₅ OH): 204.5 (20950), 206 (shoulder,

650) and 267 (11,550) nanometers.

CD (CH2OH): (O) - = 22000, (O) - = 3300.

Specific rotation [α] D = -43 ° (1%, acetone).

-8181535

PMR (CDCl ₃ ): δ 2.07 (3H, 2), 2.15 (3H, s), 3.08 (6H, s), 5.12 (4H), 5.40 (1H, s), 5.95 (1H, s) and 7.32 (10H, s).

CMR (De-acetone): δ 182.5, 172.9, 169.5, 1377.6, 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 and 20.4 p. p. m.

Molecular weight C ₃₈ H ₅₂ N ₂ O _12: Calcd: 784.3059, Found: 784.3097.

Reference Example 4

Preparation of N, N'-Bis (Benzyloxycarbonyl) -4 ', 5'-didehydro-spectinomycin (Scheme S)

To a suspension of 0.4 g of potassium hydrogen phosphate in 20 ml of anhydrous methanol is added 1.0 g of N, N'-bis (benzyloxycarbonyl) -2'-O-acetyl-4 ', 5'-didehydro-spectinone. and the resulting mixture was stirred at room temperature for 1.5 hours. The solvent was evaporated under reduced pressure and the organic portion was dissolved in 1.5% methanol in chloroform. The resulting solution was purified by chromatography on 225 g of silica gel using 1.5% methanol in chloroform as eluent. The product-containing fractions were combined and evaporated to give 0.51 g (55%) of the title compound.

CD _(CH3 0H) (0) = ^ - 8300+ 2100 (θ) 2 "= + 10500 + 2100

Specific rotation [α] θ = - 56 ° (1.0%, methanol).

CMR (D ₆ -aceton): δ 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 and 21.3 p. p. m.

Mass spectrum for tri-TMS derivative [m / e]: 814 (M ⁺ ) and 799 (M-15).

Reference Example 5

Preparation of N, N'-Bis (Benzyloxycarbonyl) -6'-hydroxy spectinomycin-3'-methylenol (Scheme SZ)

A solution of 103.9 mg (0.154 mmol) of N, N'-bis (benzyloxycarbonyl) 6'-acetoxy-spectinomycin-3'-methylenol ether in 6.8 ml of methanol was stirred with 115 mg of potassium bicarbonate at room temperature for 24 hours. The reaction mixture was evaporated, the residue redissolved in chloroform, the chloroform solution was filtered and the filtrate was evaporated again to give the title compound (90 mg). No sign of the methyl group corresponding to the acetate group was detected in the PMR and CMR spectra.

PMR (CDCl 3): δ 7.31 (s), 5.10 (s), 5-3.75 (m), 3.5 (s) and 3.1 (s).

CMR (CDCl ₃ ): δ 158, 152.5, 136.5, 128.2, 95.1, 94.9, 88.7, 77.2, 67.5, 67.2, 55.0, 34 , 1 and 29.72 ppm

Mass spectrum for the tetra-TMS derivative [m / e]: 918 (M ') and 903 (M-15).

Example 1

Preparation of Spectinomycin (Scheme T)

To a solution of 480 mg (0.8 mmol) of N, N'-bis (benzyloxycarbonyl) -4 ', 5'-didehydro-spectinomycin in 40 ml of isopropanol was added 480 mg of 10% palladium on carbon and 0.48. pyridine (5.9 mmol) was added and the reaction mixture was hydrogenated at atmospheric pressure for 5 hours. The reaction mixture was filtered and the filtrate was concentrated in vacuo and redissolved in isopropanol. To the resulting solution was added 2.0 mL (2.0 mmol) of 1 N hydrochloric acid in isopropanol and the solvent was evaporated in vacuo to give a white precipitate (440 mg). This was recrystallized from aqueous acetone to give 164 mg (0.33 mmol, 41%) of spectinomycin dihydrochloride pentahydrate. The physical and biological properties of this product are the same as those of naturally occurring products.

3c. The crystalline spectinomycin · dihydrochloride pentahydrate was also obtained in 35% yield from enol acetate prepared in Reference Example 1 and following the procedure described in the preceding paragraph.

Example 2

Preparation of 5'-DemetiI-Spectinomycin (Scheme U)

In a Parr hydrogenation vessel, a suspension of 300 mg of 10% palladium on carbon in 45 ml of isopropanol was added 333 mg (0.57 mmol) of N, N'-bis' - (benzyloxycarbonyl) -5'-demethyl spectinomycin in ml of isopropanol. followed by addition of 1.36 ml of 30N (1.36 mmol) in 1N isopropanolic hydrogen chloride solution, and the reaction mixture was hydrogenated at 2.8 at room temperature for 16 hours. The reaction mixture is filtered and the filtered catalyst is washed first with isopropanol and then with water to remove the product more effectively. The combined filtrate and washings were evaporated to dryness in vacuo and the resulting water-soluble portion was separated. Removal of the solvent in vacuo gave a white residue which was recrystallized from aqueous acetone. This gave 112 mg of the title compound as a white precipitate, m.p. 196-205 ° C (dec.).

Molecular weight based on mass spectrum of penta-TMS derivative for C ₂₃ H ₆₂ N ₂ O ₇ Si _s :

calcd: 578.3403, found: 678.3378.

CMR (D ₃ O, relative to cimetal tetramethylsilane reference material): δ 95.3, 94.9, 93.6, 71.1, 67.4, 67.0,62.9,

62.5, 61.0, 50.9, 35.9, 32.2 and 31.7.

Example 3

Preparation of 6'-hydroxy spectinomycin (Scheme V) A mixture of mg of N, N 'bis (benzyloxycarbonyl) -6'-hydroxy spectinomycin and 20 mg of palladium carbon black in 35 ml of isopropanol at 1.8 kg / cm ^{2 of} hydrogen gas After shaking for 60 hours, the catalyst is filtered off and washed with isopropanol. The filtrate was combined with the washings and concentrated to a total volume of 10 ml. To the concentrate was added 0.12 ml of 1N isopropanolic hydrogen chloride solution and the resulting mixture was evaporated. The white precipitate 65 thus obtained was redissolved in deuterium oxide to a ¹³ C separation.

-9181535 for that purpose. Isolation of the ^u C sample yielded 14 mg of material after lyophilization. This material was recrystallized from a mixture of water and steel to give the title compound, m.p. 201-205 ° C (dec.).

CMR (D ₂ O): δ 94.9,93.2, 73.6, 70.7, 67.2,64,6,62,7,60,8,

59.6, 37.0, 31.9 and 31.4 p. p. m.

Spectinomycin had an Rf of 0.5 and the title compound had an Rf of 0.25. The title compound, even in its raw form, has been shown to have antibacterial activity in a submerged disk test.

Example 4

Preparation of 4 ', 5'-Didehydro-spectinomycin dihydrochloride (Scheme W) mg of N, N'-bis (benzyloxycarbonyl) .4', 5'-didehydrospectinomycin, 70 mg of 10% palladium on carbon and 30 ml of isopropanol A mixture of 2 ml of 0.1 N isopropanolic hydrochloric acid was hydrogenated at 2.1 at 1.5 for 1.5 hours, followed by addition of 30 mg of catalyst and 1.0 ml of 0.1 N isopropanolic hydrochloric acid. , Continue for 5 hours. The reaction mixture was filtered and the filtrate was evaporated to give 50 mg of the title compound as a precipitate. This precipitate is active against Escherichia coli and does not contain spectinomycin or dihydrospectinomycin as shown by thin layer chromatography on chloroform: methanol: ammonium hydroxide (3: 4: 2) and acetone containing 3 ammonium hydroxide as a blowing agent. When tested with the latter solvent system, it gives a μγ-active spot that corresponds to a reference standard of the enantiomeric 4 ', 5'-dehydro-spectinomycin. The latter can be characterized as follows: Mass spectrum of tri-TMS derivative [m / e]: 546 (M ⁺ ), 531 (M-15), 492.426, 401, 361, 271 and 199.

CMR (D ₂ O, extreme tetramethylsilane standard): δ 189.6,

179.3,102.8, 98.5, 72.6, 69.9, 67.0, 66.5, 62.6, 61.5, 59.7,

32.1 and 22.0 p. p. m.

Reference Example 6

Preparation of N, N'-Bis (Benzyloxycarbonyl) -6'-chloro-spectinomycin (Scheme X)

At room temperature, a mixture of 200 mg (0.32 mmol) of N, N'-bis (benzyloxycarbonyl) -6'-hydroxypectinomycin and 170 mg (0.65 mmol) triphenylphosphine in 10 mL of carbon tetrachloride and 5 mL of acetonitrile was added. After stirring for 24 hours, an additional 80 mg of triphenylphosphine is added and stirring is continued for 24 hours. The reaction mixture was concentrated and the resulting oil was dissolved in chloroform (50 mL). The resulting solution was washed with 15 ml of 0.1 N hydrochloric acid and 30 ml of saturated sodium bicarbonate solution. The aqueous layer was extracted with chloroform and the combined organic layers were dried over sodium sulfate and evaporated. The crude product was then chromatographed on three 1000 micron silica gel preparative thin-layer chromatography plates, eluting with methanol: chloroform (1: 9). The silica gel was washed with warm ethyl acetate and the wash solution was collected. The filtrate is evaporated, the residue is redissolved in chloroform, the resulting solution is filtered through Celite and the filtrate is concentrated,

Yield: 87.9 mg (42%).

CM (D ₆ -aceton): 192 (C-3 'carbonyl), 157, 138.0,

129.2, 128.4, 97.3, 92.1, 75.2, 74.7, 71.5, 67.3, 66.4, 65.7,

60.2, 60.9, 57,4.47.0, 41.6 and 31.5 p. p. m.

Mass spectrum for tri-TMS derivative [m / e]: 850 (M ⁺ ), 835, 822, 814, 745 and 629.

Reference Example 7

Preparation of N, N'-bis (Benzyloxycarbonyl) -6'-bromo-spectinomycin (Scheme Y)

At room temperature, 250 mg (0.406 mmol) of N, N'-bis (benzyloxycarbonyl) -6'-hydroxymectomycin, 270 mg (1.03 mmol) of triphenylphosphine and 1 g (3.02 mmol) of carbon tetrabromide were added. of acetonitrile at room temperature

After stirring for 2.5 days, the mixture was evaporated and the residue was chromatographed on two 1500 micron silica gel preparative thin-layer chromatography plates with ethyl acetate as the eluent. The silica gel was washed with warm acetonitrile and the washings were collected. Concentration of the filtrate gave 105 mg (37%) of the title compound.

CMR (D _e -acetone): 192 (C-3 'carbonyl), 157, 138.1,

129.2, 128.4, 97.2, 92.0, 75.2, 74.5, 70.9, 67.3, 66.3, 65.6,

60.5, 57.5, 42.5, 35.5 and 31.5 p. p. m.

Mass spectrum for tri-TMS derivative [m / e]: 896, 894 (M ⁺ ), 881, 879 (M-15), 814 (M-HBr), 745 and 629.

Reference Example 8

Preparation of N, N'-Bis (Benzyloxycarbonyl) -6'-hydroxy spectinomycin (Scheme Z)

490 mg (0.64 mmol) of N, N'-bis (benzyloxycarbonyl) -3'-O-acetyl-4 '- (R) -acetoxy-6'-acetoxy-3' (5) -dihydro- A solution of spectinoinicin in acetonitrile was stirred with 1.5 g of potassium bicarbonate for 48 hours, and the reaction mixture was filtered and the filtrate was concentrated. The resulting crude product (460 mg) was dissolved in a mixture of THF (3 mL) and 4N HCl (3 mL), and the resulting reaction mixture was diluted with chloroform (200 mL) and the organic layer was washed with saturated sodium bicarbonate solution and water. dried over sulfate and evaporated to give 330 mg of product. Thin layer chromatography and CMR showed a purity of at least 85-90%. The product was identified as the title compound by CMR, mass spectrometry, and HPLC.

Example 4

Preparation of 6'-Chloro-spectinomycin and its dihydrochloride salt (Scheme AA)

To a solution of 43.4 mg (0.068 mmol) of N, N'-bis (benzyloxycarbonyl) -6'-chloro-spectinomycin in 5.5 ml of isopropanol is added 42 mg of 10% palladium-on-metal palladium catalyst. to vacuum several times, purge with nitrogen gas between each treatment, and vacuum treatments were repeated, with hydrogen gas being purged occasionally.

After stirring for 1 hour under a hydrogen atmosphere of -10181535 g, it was diluted with 25 ml of isopropanol and filtered through a pad of Celite. The filtrate was concentrated to about 10 ml and the concentrate was acidified with hydrochloric acid in isopropanol and evaporated to dryness. The resulting solid was dissolved in water (4 mL), filtered and lyophilized. The crude product (30 mg) was further precipitated from a mixture of water and acetone to give the title compound (7 mg, 23.5%).

CMR (D ₂ O): 97.4, 94.5, 92.9, 72.3, 71.0, 66.8, 62.7, 60.7,

59.8, 47.5, 38.3, 31.9 and 31.4.

Mass spectrum for tri-TMS derivative [m / e]: 581 (M-1), 566, 551, 545 (M-HCl).

Example 5

Preparation of 6'-bromo-spectinomycin and its dihydrochloride salt (Scheme AB)

To a solution of N, N'-bis (benzyloxycarbonyl) 6'-bromo spectinomycin (29.9 mg, 0.044 mmol) in isopropanol (3.0 mL) was added palladium catalyst (10 mg, 10% metal) and the resulting reaction mixture. and stirred under a hydrogen atmosphere at atmospheric pressure for 3 hours. The reaction rate is reduced, so the catalyst is filtered off and the reaction is re-started after adding another 65 mg of fresh catalyst and after 2 hours an additional 70 mg of fresh catalyst. After stirring for an additional 1 hour, the reaction mixture was filtered through Celite and the filtrate was concentrated to one third of its original volume. The concentrate was acidified with 0.15 ml of normal isopropanolic hydrochloric acid and evaporated to dryness. The crude precipitate was dissolved in water (3 mL) and the resulting solution was filtered and the filtrate was lyophilized. 17 mg (79%) of the dihydrochloride salt of the title compound are obtained.

CMR (D ₂ O): 94.7, 94.6, 92.8, 71.6, 71.0, 67.1, 66.8, 62.8,

60.8, 59.8, 39.3, 36.3, 31.9 and 31.5p. p. m.

Claims (2)

Patent claims A process for the preparation of a compound of formula (I): wherein: R (IX) or (X), wherein R j is hydrogen or C 1-4 alkyl, halo (C 1-4) alkyl, or hydroxy (C 1-4) alkyl, but other than methyl, while R'j is alkyl of 1-4 carbon atoms, R ₃ and R ₃ each independently represent an alkyl group containing from 1 to 4 carbon atoms and pharmaceutically acceptable salts thereof, characterized in that j) a compound of formula VI wherein R ₂ and R ₃ are as defined herein, while T is a protecting group such as aralkoxycarbonyl or optionally halogenated alkoxycarbonyl, preferably benzyl &gt; -xycarbonyl, with a compound of formula VII, wherein R 'is the same as R but with formula X other than R 1 and R 1 are hydroxyalkyl protected with an acyl group and R ₆ is alkyl having 1 to 4 carbon atoms or an acyl protecting group, preferably acetyl, to give a compound of formula III wherein R ₂ and R ₃ are while T, R 'and Rj _e are as defined herein - d and then deprotecting a compound of formula (II) wherein R, R ₂ and R ₃ are as defined herein, or T is as defined herein, in a known manner, or ( ₂ ) a compound of formula (II) Wherein R, R ₂ , R ₃ and T are as defined in process (j), and are optionally cleaved and, if desired, converted to a pharmaceutically acceptable salt of the compound of formula (I) prepared according to process ( _a ) or ( ₂ ). 2. A process for the preparation of pharmaceutical compositions, characterized in that a compound as claimed in claim 1, j) or a compound of formula prepared by the procedure ₂₎ (I) - wherein R (IX) or (X), wherein R j is hydrogen or C 1-4 alkyl, halo (C 1-4) alkyl, or hydroxy (C 1-4) alkyl , but other than methyl, while R'j is alkyl of 1-4 carbon atoms, R ₂ and R ₃ each independently represent a C 1-4 alkyl group - or a pharmaceutically acceptable salt thereof, in a known manner, in admixture with carriers and / or excipients conventionally used in the manufacture of a medicament.