HU176355B - Process for preparing 3',4'-dideoxy-kanamycin derivatives - Google Patents

Abstract

PURPOSE:To obtain title compound through fewer reaction steps by hydrogenation of 3',4'-dideoxy-3'-enokanamycin B derived from kanamycin as starting material.

Description

BACKGROUND OF THE INVENTION The present invention relates to a novel process for the preparation of the antibiotic 3 ', 4'-dideoxy-kanamycin B, which is active against various Gram-positive and Gram-negative bacteria, in particular kanamycin-resistant microorganisms such as kanamycin-resistant Staphylococci and kanamycin-resistant Escherichia. can be used to combat diseases caused by coli.

The structure of the 3 ', 4'-dideoxy-kanamycin B is represented by Structure I ^1st According to the patent specification U.K. No. 1349302 to protect the starting amino groups of kanamycin B such that a Schiff base are transformed into type groups A and the 3 'and 4'-position and the 4 "- and ^one 6' -position the hydroxyl groups present in each pair are converted into acetal or ketal. Subsequently, an alkanoyl or aroyl group is used to temporarily protect the 2 'hydroxyl group and then selectively deprotect the 2', 3 'and 4' hydroxyl groups with dilute acid. They then sulfonylate the hydroxyl groups at the 3'- and 4'-positions, form a double bond between the 3'- and 4'-carbon 2-atom using an alkali metal bromide or iodide, and then hydrogenate the double bond. The acidic group is then deprotected at the 4 ' and 6 ' position and then cleaved with the remaining protecting groups to give 3 ', 4 ' -dideoxycannamycin B.

The disadvantage of the above method is that it requires complex operations for the temporary protection of the amino and hydroxy groups and subsequent cleavage of the protecting groups and provides DKB with a poor overall yield of only 10%.

The present invention relates to a novel process for the preparation of 3 ', 4'-dideoxy-kanamycin B which is more advantageous than the methods known hitherto in that it does not require the use of alkali metal bromides or iodides and zinc dust, but less expensive reagents. Similarly to the previously known method, we start from kanamycin B, but swallow 3 ', 4'-dideoxy-kanamycin B via a new pathway.

The present invention relates to a. a process for the preparation of 3 ', 4'-dideoxy-kanamycin B and its acid addition salts by:

1. Compounds of formula li, wherein R is hydrogen or C 1-6 alkyl, Z is cyclohexylidene, and 3 ', 4'-epoxy can be in the a- or < 1 > position, and is reacted with xanthate. the 3 ', 4'-dideoxy-3'-canamycin B derivative of the general formula wherein R and Z are as defined above and the formula IV wherein R and Z are as defined above and the can be in the - position to give the 3 ', 4'-epithiocannamycin B derivative,

2. recovering the 3 ', 4'-dideoxy-3'-enamen-kanamycin B derivative from the reaction mixture,

3. Cleavage of the 3 ', 4'-dideoxy-3'-canamycin B derivative obtained in this manner by conventional methods of protecting the amino-protecting group -COOR and the hydroxy protecting group Z to obtain the 3' , 4'-dideoxy-3'-en-kanamycin B, then

4. Hydrogenation of the compound thus obtained in a conventional manner to give 3 ', 4'-dideoxy-kanamycin B and, if necessary, converting the product into an acid addition salt thereof.

According to a preferred embodiment of process a) of the present invention, the first step of the above reaction sequence, that is, the reaction of the compounds of formula II with an xanthate in an organic solvent, preferably at a temperature of 50 ° C to 100 ° C. Preferably, a lower alcohol such as methanol or ethanol is used as the organic solvent. The xanthate used for this treatment is of the formula R'OCSSMe, wherein R 'is a lower alkyl group and Me is an alkali metal such as sodium or potassium. In this reaction, the 3 ', 4'-dideoxy-3'-ene derivatives of the general formula III and the 3', the by-product of the general formula IV are generally used as the main product by washing the reaction mixture with water, recovering the solvent by distillation and distilling the residue to dryness. 4'-epithio derivatives were obtained.

The mechanism of the reaction between the 3 ', 4'-epoxy derivatives of formula II and xanthate has not been elucidated yet, but it is likely that the 3', 4'-epoxy derivatives are converted via the epithio derivatives of formula IV 3 ', 4'-dideoxy-3'-enan-kanamycin B derivatives of formula III.

Recovery of the product from step 1, i.e. step 2, can be carried out, for example, preferably by chromatography on a silica gel plate, for example, using a 1: 1 by volume mixture of carbon tetrachloride and acetone.

In a further preferred embodiment of process a) according to the invention, the third step is carried out in the usual manner, the removal of the amino protecting groups COOR and the hydroxyl protecting group Z. For example, the hydroxyl protecting group Z may first be removed by mild hydrolysis with dilute hydrochloric acid or aqueous acetic acid, followed by hydrolysis with barium hydroxide or hydrogenolysis in the presence of palladium as a catalyst.

According to a further preferred embodiment of process a) according to the invention, the fourth step, the hydrogenation reaction, can be carried out in known manner. Thus, known hydrogenation catalyst preferably comprising a platinum group metal such as platinum or palladium, _(hydrogen in the reaction may be carried out. It is also used a Raney nickel for this purpose.

The advantages of process a) of the present invention, as described above, over the prior art process mentioned above, are as follows:

- it is possible to form an unsaturated bond between the carbon atoms 3 'and 4' without resorting to a reaction requiring alkali metal bromide or iodide and zinc dust,

- in the final step, the final intermediate to be converted to the final product, 3 ', 4'-dideoxy-canamycin B, is 3', 4'-dideoxy-3'-enam-kanamycin B, i.e. no longer having any protecting groups To get a compound of formula V,

much less impurity is formed in the final reaction step, catalytic hydrogenation, and the reaction is relatively easy to carry out and therefore requires only less purification of the final product than in the prior art where the protecting groups on the amino and hydroxy groups are hydrogenated. is removed.

It should be noted, however, that if necessary, steps 3 and 4 of process a) of the present invention can be performed in reverse order, similarly to the procedure known in the literature.

If desired, the free base product, 3 ', 4'-dideoxy-kanamycin B, can be converted into its acid addition salt with inorganic or organic acids. For example, the sulfate of 3 ', 4'-dideoxy-kanamycin B can be obtained by adjusting the pH of the aqueous solution of the free base to 6.8 by adding dilute sulfuric acid, filtering the solution, filtering the carbon and freeze-drying the filtrate. .

It has further been found that the 3 ', 4'-epithio derivatives of the general formula IV, which are formed as a by-product of the process of step a) of the present invention, can also be used to prepare 3', 4'-dideoxy-kanamycin B. It has been found that there are two ways of converting the 3 ', 4'-epithio derivatives of the general formula IV into one of the known intermediates for the preparation of 3', 4'-dideoxy-kanamycin B; by acidic treatment, these compounds are converted to the 3 ', 4'-dideoxy-3'-canamycin B derivatives of the general formula V, and the other method consists of hydrazine or Raney's derivative of the 3', 4'-epithio derivatives. nickel is used to prepare the 3 ', 4'-dideoxy-3'-canamycin B derivatives of formula III.

The present invention also relates to process b) for the preparation of 3 ', 4'-dideoxy-kanamycin B and its acid addition salts by:

1. treating a compound of formula II wherein R and Z are as defined above and the 3 ', 4'-epoxy group is at the' 5 'position with xanthate to form a 3', 4'-epithio of formula IV derivatives wherein R and Z are as defined above and the 3 ', 4'-epithio group may be in the a or 0 position; and the 3' of formula III wherein R and Z are as defined above. , 4'-dideoxy-3'-canamycin B derivatives,

2. recovering the 3 ', 4'-eptothiocanamycin B derivatives from the reaction mixture,

3. Acidic treatment of the 3 ', 4'-epithio derivatives thus obtained yields 3', 4'-dideoxy-3'-enen-kanamycin B of formula V and

4. Conventional hydrogenation of the compound thus obtained to give 3 ', 4'2

-dideoxycannamycin B and, if necessary, converting the product into an acid addition salt thereof.

Steps 1, 2 and 4 of process b) according to the invention correspond to steps 1, 2,

2 and 4, as shown above.

Step 3 of process b) of the present invention, acid treatment of the 3 ', 4'-epithio derivatives of formula IV, is preferably carried out in a lower alkanol, such as methanol or ethanol, as a solvent, a hydrochloric acid such as concentrated hydrochloric acid or hydrobromic acid. In general, however, a non-oxidizing mineral acid such as sulfuric acid may be used for this treatment. Preferably, the reaction is carried out at a temperature of 0 ° C to 30 ° C.

The present invention also relates to process c) for the preparation of 3 ', 4'-dideoxy-kanamycin B and its acid addition salts by:

1. derivatives of formula II of the 3 ', 4'-epoxy-kanamycin B protected amino and hydroxy groups, wherein R and Z are as defined above, treated with a xanthate of formula IV, wherein R and Z are as defined above, and the 3 ', 4'-epithio group may be in the a or 0 position for the 3', 4'-epithiocannamycin B derivatives and the 3 ', 4' of the formula III wherein R and Z are as defined above. The '-dideoxy-3' -en-kanamycin B derivatives are then obtained

2. treating the 3 ', 4'-epithio derivatives of formula IV with S' ^ '- dideoxy-S'-enan-kanamycin B derivatives of formula III with hydrazine or Raney nickel;

The reaction mixture was obtained after step 1 and then

3. Removal of the amino protecting groups -COOR and the hydroxyl protecting groups Z by conventional methods from the 3 ', 4'-dideoxy-kanamycin B derivatives obtained in the above manner to give 3', 4'-dideoxy-3 'of formula V -en-kanamycin B was obtained and finally

4. hydrogenating the compound obtained in a conventional manner to give 3 ', 4'-dideoxycannamycin B and, if necessary, forming an acid addition salt thereof.

Steps 1, 3 and 4 of process c) according to the invention correspond to steps 1, 3,

3 and 4, as shown above.

The c. According to a preferred embodiment of the process, the hydrazine treatment is preferably carried out in a lower alcohol such as methanol or ethanol as solvent, with hydrazine, especially its hydrate, at room temperature, generally between 15 ° C and 25 ° C. The amount of hydrazine used is preferably 10 to 30 times molar with respect to the compounds of formula IV. If Raney nickel is used for Step 2, it is preferable to dissolve the compounds of Formula IV in a lower alkanol, such as methanol, by adding an appropriate amount of, for example, three times the weight of the compound of Formula IV to Raney Nickel. solution, then

The reaction mixture is stirred for 1 to 3 hours at room temperature, generally between 15 ° C and 25 ° C.

According to process c) according to the invention is ₅ In another preferred embodiment of the second operation can be performed if necessary also in a way, that beside the 3 ', 4'-dideoxy-3'-en-kanamycin B derivatives of general formula III obtained from IV formula 10 3 ', 4' -epitio derivatives starting material, but in general, the use of particularly large quantities, preferably, obtained directly upon completion of the operation 1), the compounds of formula III and IV, a reaction mixture containing a side ₁₅ as starting material.

It has also been found that 3 ', 4'-dideoxy-3'-enanthamycin B of formula V can be prepared from 3', 4'-epoxy derivatives of formula II, also by another reaction route, in high total yield. This reaction path is as follows:

1. II - wherein R and Z are as defined above - is obtained 3 ', 4'-epoxy-derivative of an acylating agent, for example by treatment with benzoyl chloride, that is at the 2-position hydroxyl group is acylated in a conventional manner to ₂₅ Compounds of formula VI wherein R and Z are as defined above and Y is acyl, then

2 from the above Formula ₃ Q is a compound of formula VI, an alkali metal or alkaline earth metal iodide such as sodium iodide, preferably sodium acetate and treated with acetic acid in the presence of the formula VII - wherein R, Z and Y are as defined above compounds of the invention

3. sulfonylation of the 3'-hydroxy group of the compounds of formula VII obtained above with a sulfonylating agent such as mesyl chloride, tosyl chloride or benzylsulfonyl chloride, preferably in pyridine as solvent at a temperature below 10 ° C. wherein R, Z and Y are as defined above and W is a mesyl, tosyl or benzylsulfonyl group;

4. Conversion of the reaction mixture obtained in Step 3 above, preferably at a temperature of 80 to 100 ° C, converts the compounds of formula VIII to the compounds of formula IX wherein R, Z and Y are as defined above, and finally

5. Compounds of formula IX obtained in the above manner are obtained by treatment with an alkali metal or alkaline earth metal alcoholate such as sodium alcoholate in a manner known per se, 3 ', 4'-dideoxycannamycin B derivatives.

⁵⁵ X - wherein the R and Z are as defined above - an amino group at the 4 - protected hydroxyl group at the "6" 4ielyzetben 3 ', 4'-0-epoxy-kanamycin B derivatives, which according to the invention ) and (c) a starting material for process ( ⁶⁰ ), also a novel compound, is also included within the scope of the invention.

The preparation of the 3 ', 4'-epoxy-kanamycin B derivatives of formula X according to the present invention is accomplished by the following reaction steps starting from kanamycin B.

First, the amino groups of kanamycin B are protected in a known manner. For this purpose, a chloroformic acid ester of the formula ROCOC1, where R is as defined above, for example methyl, is reacted with kanamycin B, the five amino groups of which are thus converted to the urethane group of the formula -NHCOOR, as disclosed in Japanese Patent Application 7595/75. Thus, the penta-N-protected-kanamycin B derivatives of formula XI, wherein R is as defined above, are obtained.

The next step also protects the 4 "and 6" hydroxyl groups in a manner known per se. To this end, compounds of formula XI are reacted with a known reagent commonly used to protect hydroxyl groups, such as an alkylidene, arylidene, cyclohexylidene or tetrahydropyranylidene. Examples of suitable reagents for protecting hydroxyl groups are acetaldehyde, 2 ', 2'-dimethoxypropane, anisaldehyde, benzaldehyde, dimethylacetal, tolualdehyde, 1,1-dimethoxycyclohexane and 1,1-dimethoxytetrahydropyran. Preferably, the reaction is carried out in a polar organic solvent, such as dimethylformamide, in the presence of a catalytic amount of p-toluenesulfonic acid, at room temperature, for example between 15 ° C and 25 ° C, for 15-20 hours. In this way, the hydroxyl protecting group selectively enters into the hydroxyl groups at the 4 ' and 6 ' positions and the formula XII - wherein R is as defined above and Z is alkylidene, arylidene, cyclohexylidene or tetrahydropyranylidene. - 4 ", 6" -O-protected derivative. It is important that the temperature of the reaction mixture is maintained below 30 ° C during this process, since at higher temperatures the reactant may also attack the hydroxyl groups at the 3'- and 4'-positions.

Subsequently, the compounds of formula XII are subjected to an acylation reaction to protect the hydroxyl groups in the 2 "and 3" positions with an acyl-type hydroxyl group. This is usually accomplished by dissolving the compounds of formula XII in pyridine, adding an acylating agent such as an acid chloride at low temperature, preferably below 5 ° C, and stirring the reaction mixture for several hours.

Preferred acylating agents for this purpose are, for example, chlorides of C 2-4 alkanoic acids such as acetyl chloride or aroyl chlorides such as benzoyl chloride. Most preferably, benzoyl chloride is used. During acylation at temperatures below 5 ° C, the hydroxy groups 41 and 5 do not react because they are relatively less reactive. In this way, the compound of formula XIII, wherein R and Z are as defined above, and X and Y are each an acyl group such as an alkanoyl group, especially a lower alkanoyl group such as acetyl, or an aroyl group such as benzoyl 3'-diacyl derivatives were obtained.

If, for certain purposes, the 2 ' -monoalkylated derivatives, i.e. compounds of formula XIII, wherein X is hydrogen, are required, they may be prepared by performing the acylation step under mild conditions. For example, when benzoyl chloride is used, the 2 'monobenzoyl derivative can be obtained as the main product by slowly adding benzoyl chloride in small portions at temperatures below 0 ° C. On the other hand, the addition of benzoyl chloride at 0 ° C to room temperature, preferably 0 ° C to 5 ° C, yields, at least as the main product, the 2 ', 3'-dibenzoyl derivative. If this step results in a mixture of 2'-monoacylated and 2 ', 3'-diacylated compounds, the two products can be separated by chromatography in a manner known per se, such as silica gel plates, chloroform: methanol (2: 1).

The 4'-hydroxyl group of the compounds of formula XIII is then sulfonylated to give the compound of formula XIV wherein R, Z, X and Y are as defined above and W is a mesyl, tosyl or benzylsulfonyl group. 4'-O-sulfonyl derivatives are obtained. Preferably, this process is carried out by reacting the compounds of formula XIII with pyridine in mesyl chloride, tosyl chloride or benzylsulfonyl chloride. The 4'0-sulfonylation is carried out at temperatures up to 50 ° C. Most preferably, mesyl chloride is used.

Thereafter, the 4'-O-sulfonylated derivatives of formula XIV are treated with a metal alcoholate to give the aforementioned 3 ', 4' - / 3-epoxy derivatives of formula X. Preferably, the epoxide is prepared by dissolving a compound of formula XIV in a solvent such as water, a lower alkanol such as methanol or ethanol, diethylene glycol dimethyl ether, sulfolane, tetrahydrofuran or dimethyl sulfoxide. to a solution of a metal alcoholate, generally an alkali metal or alkaline earth metal alcoholate, such as sodium potassium or lithium alcohol, in particular a lower alcohol such as sodium ethylate or sodium methylate, and the solution obtained in a suitable manner, 1-3. The reaction mixture is maintained at room temperature, usually between 15 and 25 ° C, for one hour. The alkaline medium used in the epoxide-forming reaction also cleaves the protecting group Y of the hydroxyl group at the 2 '' position, thereby releasing the hydroxy group at the 2 'position in the 3', 4 'epoxy derivatives of formula X.

3 ', 4' -? - epoxy derivatives corresponding to the 3 ', 4' -? - epoxy derivatives of the general formula AX from the 3'-O-tosyl derivatives corresponding to the 4'-O-sulfonylated derivatives of the general formula XIV, i.e. can be obtained from compounds of formula XIV wherein W is hydrogen and X is a tosyl group by the method described above for treating the starting materials with an alkali metal alcoholate such as sodium 4-methylate. This method is described in detail in German Patent Publication No. 2,555,479.

In addition, Scheme Z of the drawings and described below summarizes 3 ', 4'-dideoxy-kanamycin B according to the present invention starting from kanamycin B and in a novel intermediate, 3', 4'-O- of formula Xa. processes for the preparation of said intermediate via epoxy derivatives, wherein said intermediate is converted to the desired product by certain steps of the process of the invention.

Reaction Scheme Z is a temporary protection of five amino groups with a group of formula -COOR

I ----------------------------------------------- xi a Temporary protection of hydroxyl groups at the 4 ' and 6 ' position by one of the Z groups, Sulfonylation of the hydroxyl group at the 2 ' and 3 ' Formation of a 4'-epoxide group by means of a metal alcoholate

XII

XIII

XIV

Reaction Xa with xanthate (Methods a, b and c) with hydrazine or

With nickel Raney (method c)

Note: d. Process A is an alternative embodiment of process a and b of the invention.

The invention is further illustrated by the following non-limiting examples.

Example 1

1. Preparation of penta-N-ethoxycarbonyl-kanamycin B 60

Penta-N-ethoxycarbonyl-kanamycin B was prepared from the free base of canaryiden B using the method described in Example 1 of British Patent No. 1,349,302. 65

2. Preparation of 2 "3'-diO-benzoylpenta-N-ethoxycarbonyl-4", 6 "-O-cyclohexylidenecanamycin B In 10 ml of dimethylformamide, 10 g of penta-N-ethoxycarbonycanamycin B are mixed. p-toluenesulfonic acid is added until the pH drops below 3 and 10 ml of cyclohexanone dimethyl ketal are added and the mixture is stirred for 18 hours at 25 ° C. TLC (Merck silica gel, developing solvent: chloroform: methanol = 10: 1, v / v) was carried out to check that the reaction was complete and neutralized with triethylamine. The neutralized mixture was concentrated under reduced pressure to a total volume of 25 ml and the concentrate was dissolved in 150 ml of pyridine. The solution is cooled to 0 ° C to 5 ° C, benzoyl chloride (3.9 ml) is added and the reaction mixture is maintained at this temperature for 3 hours. ⁵ Check by TLC that the reaction is complete. Water (5 mL) was added to the reaction mixture, stirred at room temperature for half an hour, concentrated and poured into water (200 mL). The precipitate was filtered off, yield 10: 12.7 g (95%). Chromatography on silica gel gives the pure material the physical constants: [α] ^D = 76.6 ° (c = 1, pyridine), m.p. 233-235 ° C.

Analysis for C _{s 3} H _{7 3} N ₅ O _{2 2} 15

date: C = 56.22% N = 6.19% H = 6.51% found: C = 55.98% H = 6.44%, N, 5.60%. 20

3. The 2 ', 3'di-O-benzoylpenta-N-ethoxycarbonyl-

Preparation of 4 ", 6" -di-O-Cyclohexylidene-4'-O-Mesyl-Kanamycin B

Dissolve 5 g of 2 "3'-di-O-benzoylpenta-N-ethoxycarbonyl-4", 6 "-di-O-cyclohexylidenecannamycin B in 100 ml of addin, add 1.4 ml of mesyl chloride and The reaction mixture was stirred at 40 ° C. After cooling to room temperature, water (5 mL) was added to quench the excess mesyl chloride and the solution was concentrated. Add 200 ml of water to the residue which precipitates and is filtered off. Yield: 5,0g (94%), [α] ρ = ⁵ 103.6 ° (c = 1, pyridine), m.p .: 35176-179 ° C.

Analysis for C H ₇₅ N ₅ O2 _{S4 4} S Calcd Calculated: C, 53.59; H = 6.25% N, 5.79% S = 2.65% Found: C, 53.28; H = 6.25% N = 5.41% S, 2.95%.

4. The 3 ', 4' - / 3-epoxy-penta-N-ethoxycarbonyl-

Preparation of 4 ", 6" -O-Cyclohexylidenecanamycin B 45

5 g of 2 ", 3'-di-O-benzoylpenta-N-ethoxycarbonyl-4", 6 "-O-cyclohexylidene-4'-O-mesyl-kanamycin B are dissolved in 100 ml of methanol and 2.2 g of sodium methylate are added. . 2 hours at room temperature stirring to ₅₀ while cooling the mixture, thin layer chromatography on silica gel (eluent: 1 in volume: carbon tetrachloride: acetone 1) indicated that the reaction was complete or not. Cooled in ice, the crude reaction mixture, 1.25 ml of concentrated hydrochloric acid ₅₅ was neutralized and concentrated. The desired product precipitates from the neutralized concentrate to add 100 ml of water and isolates by filtration. Yield: 3.5 g (95%), [α] ⁵ θ = 37.8 ° (c = 0 pyridine), m.p .: 254-258 ° C (decomposition with foaming). ₆₀

Analysis for C ₃₉ H ₆₃ N _{9 s} Oi Found Calculated: C, 51.69; H = 7.02% N = 7.73% O = 33.55% Found: C, 51.86; H = 6.89% N, 7.58% O = 33.67%.

5. Preparation of 3 ', 4'-dideoxy-3'-ene-penta-N-ethoxycarbonyl-4', 6 '-O-cyclohexylidenecanamycin B in ml n-butanol was mixed with 800 mg of 3', 4'-βe ρ oxi-penta-N-ethoxycarbonyl-4 ", 6" -O-cyclohexylidenecannamycin B, 1.7 g of potassium n-butyl xanthate are added to the suspension and the mixture is stirred at 80 ° C for 4 hours. . After confirming (TLC on silica gel plates, 1: 1 carbon tetrachloride / acetone) that the reaction was complete, the reaction mixture was cooled and washed with water (2 x 40 mL). The butanol solution is distilled to dryness and the residue weighs 900 mg. This product was found to be 3 ', 4'-dideoxy-3'-penta-N-ethoxycarbonyl4', 6 '-0-cyclohexylidenecanamycin B and 3', 4'-epithio by TLC on silica gel. penta-N-ethoxycarbonyl-4 ", 6" -O-cyclohexylidenecannamycin B ca. 1: 1 mixture,

The 3 ', 4'-dideoxy-3'-ene-penta-N-ethoxycarbonyl-4', 6 '-cyclohexylate was chromatographed on silica gel plates with a 5: 1 mixture of carbon tetrachloride and acetone as the eluent. Bt Ι [α] θ ^δ = 24.7 ° (c = 1.0, methanol) | and 3 ', 4'-penta epitio-N-ethoxycarbonyl-4 ", 6" -O-cyclohexylidene-kanamycin Bt I [a] ^ ⁵ = 10.8 ° (c = 0, water) mp. 250-260 ° C (dec.).

Analysis of this compound C ₃₉ H ₆₃ N _s O ₁₈ S

Found date: C = 50.79% H = 6.90%, N = 7.60% S = 3.48%, found: C = 50.41% H = 6.95% N = 7.45% S, 3.48%.

6. Preparation of 3 ', 4'-dideoxy-3'-enan-kanamycin B Dissolve 475 mg of 3', 4'-dideoxy-3'-penta-N-ethoxycarbonyl4 "-6" -O in ml of methanol. -cyclohexylidene-kanamycin B and then 1 N hydrochloric acid was added until the pH was 2.0. After half an hour at 50 ° C, water (5 ml) and barium hydroxide octahydrate (1.4 g) were added, methanol was distilled off from the mixture, refluxed for 8 hours and finally cooled. Passing carbon dioxide gas into the cooled mixture, the barium carbonate precipitates and is filtered off. Purification on a column packed with ammonium phase Amberlite CG-50 ion exchange resin afforded the desired compound (560 mg, 26%).

Analysis calculated for C ₁₈ H _{3 S} N ₅ O ₈ : C, 48.11;

N, 15.59. Found: C, 47.85;

N, 15.40%.

H = 7.80%

H = 7.95%

7. Preparation of 3 ', 4'-dideoxy-kanamycin B Dissolve 120 mg of 3', 4'-dideoxy-3'-kanamycin B in 120 ml of water, add 0.2 ml of Raney nickel and place at room temperature for 2 hours. hydrogen at atmospheric pressure is passed through the solution. The catalyst is then filtered off and the filtrate is distilled to dryness. Yield: 108 mg (95%).

Example 2

1. Preparation of penta-N-tert-butoxycarbonyl-kanamycin B To a mixture of ml of water, 24 ml of triethylamine and 48 ml of dimethylformamide is added 10 g of kanamycin B, and 40 g of monothiosic acid half-butyl are added at room temperature. -ester S-half-(4,6-dimethylpyrimidin-2-yl) ester and stirred for 18 hours at the same temperature. Water (150 mL) was added, the precipitated crystals were filtered off, and washed thoroughly with water saturated with ethyl acetate. Yield: 20 g (100%), m.p. 229-234 ° C (decomposes with foaming).

Analysis calculated for C43H ₇₇ N _s 02o O: C = 52.47%, H 7.90%,

N, 7.12; Found: C, 52.61; H, 7.86;

N, 6.93%.

2. Preparation of 3 ', 4' - / 3-epoxy-penta-N-tert-butoxycarbonyl-4 ", 6" -O-cyclohexylidenecannamycin B

4. Preparation of 3 ', 4'-dideoxy-3'-enan-kanamycin B Dissolve 1.15 g of 3', 4'-a-epi5 thiopenta-N-tert-butoxycarbonyl-4 'in methanol. -6 "-O-Cyclohexylidenecanamycin B, 3 mL of concentrated hydrochloric acid was added and the mixture was stirred at room temperature for 3 hours.

Distill the solution to dryness, dissolve the residue in 18 ml of water and adjust the pH of the solution to 6.5 by adding 2.5 ml of normal sodium hydroxide solution. Is slowly added into the solution phase of ammonium Amberlite CG-50 ion exchange resin column of 20 ml and then washed with water for 1 ₅ column, then eluted with 0.1 N and 0.3 N ammonium hydroxide. Concentration of the eluate affords the desired material. Yield: 194 mg.

20o 5. Preparation of 3 ', 4'-dideoxy-kanamycin B Dissolve 120 mg of 3', 4'-dideoxy-3'-enan-kanamycin B in ml of water, add 0.2 ml of Raney nickel and place at room temperature for 2 hours. hydrogen at atmospheric pressure is passed through the solution. The catalyst is filtered off and the filtrate is distilled to dryness. Yield: 108 mg (90%).

Example 3

Following the procedure of Example 1, Part 2, 3 and 4, and starting from 10 g of penta-N-tert-butoxycarbonyl-kanamycin B was the desired product, yield 80%, [α] θ ³ = 27 ° (c = 1.0 pyridine), m.p. 232-234 ° C (decomposes with foaming).

3. Preparation of 3 ', 4'-epithiopenta-N-tert-butoxycarbonyl4 ", 6" -0-cyclohexylidenecannamycin B

In 100 ml of n-butanol was added 10 g of 3 ', 4', 3-epoxy-penta-N-tert-butoxycarbonyl-4', 6 '-O-cyclohexylidenecannamycin B, and 9.5 g of potassium nitrate was added to the suspension. butyl xanthate and kept at 90 ° C for 2 hours. After completion of the reaction, the reaction mixture was cooled, washed with water (2 x 100 mL) and the butanol solution was distilled to dryness. Thus, 3 ', 4'-epithiopenta-N-tert-butoxycarbonyl-4', 6 '-O-cyclohexylidenecannamycin B and 3', 4'-dideoxy-3'-ene-penta- A mixture of N-tert-butoxycarbonyl-4 ", 6" -O-cyclohexylidenecanamycin B is obtained. Production: llg. The crude product was chromatographed on silica gel (eluent: chloroform: methanol = 50: 1) to give 3.9 g (35%) of 3 ', 4'-epithiopenta-N-tert-butoxycarbonyl 4', 6 '-O-cyclohexylidene. kanamicm Bt I [a] p ⁵ = 23 ° (c = 0, pyridine), m.p .: 235-238 ° (decomposition with foaming) | win.

Analysis for C ₄₉ H ₈₃ N _s O _{1 R} S

date: C = 55.39%, N, 6.59% H = 7.89% S = 3.02% found: C = 55.10% H = 7.94%, N = 6.31% S, 3.30%.

Preparation of 3 ', 4'-epithiopenta-N-ethoxycarbonyl-4', 6'-O-cyclohexylidenecannamycin B Dissolve 100 mg in ml pyridine as disclosed in German Patent Publication No. 2,555,479. 3 ', 4'-α-epoxy-penta-N-ethoxycarbonyl-4', 6 '-O-cyclohexylidenecanamycin B obtained as described in Example 1A, 100 mg of potassium ethyl xanthate were added and the mixture was refluxed for 1.5 hours. the mixture is then distilled to dryness. A 2: 3 mixture of water and chloroform is added to the residue, the chloroform phase is separated off, washed three times with 20 ml of water and the solvent is distilled off. This gave 70 mg of crude product of 3 ', 4'-epithiopenta-N-ethoxycarbonyl-4', 6 '-O-cyclohexylidenecanamycin B and 3', 4'-dido xi-3. Mixture of '-en-penta-N-ethoxycarbonyl-4', 6 '-O-cidohexylidenecanamycin B.

The components of the mixture were separated by thin layer chromatography as described in Example 1 (5) above.

Example 4

1. Preparation of 3 ', 4'-dideoxy-3'-ene-penta-N-ethoxycarbonyl-4', 6 '-O-cyclohexylidenecannamycin B ⁶⁰ ml of methanol are dissolved in 470 mg. example

3'4'-epithiopenta-N-ethoxycarbonyl-4'6'O-cyclohexylidenecannamycin B obtained as described in Example 5 or Example 1 (1), 0.22 ml of hydrazine hydrate are added and and allowed to stand at room temperature for 2 hours. The mixture was then distilled to dryness and treated with water (20 mL). The precipitate was filtered off, yield 362 mg (80%). ^[6 αβ 24.7 ° (c = 1.0, methanol).

Analysis calculated for C ₃₉ H _{6 S} N _{s 8} Oi O: C = 52.50%, H 7.36%,

N, 7.85. Found: C, 52.31; H, 7.53;

N, 7.49%.

2. Preparation of 3 ', 4'-dideoxy-3'-enan-kanamycin B Dissolve 475 mg of 3', 4'-dideoxy-3 '-ene-penta-N-ethoxycarbonyl-4' in ml of methanol. '-O-cyclohexylidenecannamycin B and treated as described in Example 1, paragraph 6 to give the desired product.

3. Preparation of 3 ', 4'-dideoxy-kanamycin B

3 ', 4'-Dideoxy-3'-enan-kanamycin B is exemplified in Example 1

Treatment as described in point 7 gives the desired product. The compound monocarbonate had an Rf of 0.19 in n-butanol (ethanol) chloroform (17%) ammonium hydroxide (4: 4: 2: 3).

Example 5

1. Preparation of 3 ', 4'-dideoxy-3'-ene-penta-N-tert-butoxycarbonyl-4', 6 '-O-cyclohexylidenecanamycin B: Dissolve 585 mg of methanol in ml of Example 2.

3 ', 4'-epithiopenta-N-tert-butoxycarbonyl4 ", 6" -0-cyclohexylidenecanamycin B obtained as described in point 3, add 500 mg of Raney nickel R-100, and stirred for 2 hours at room temperature (20 ° C). The Raney nickel was then filtered off and the filtrate was distilled to dryness to give 450 mg (80%) of product.

2. Preparation of 3 ', 4'-dideoxy-kanamycin B

In the same manner as in Example 1, points 6 and 7, 450 mg of 3 ', 4'-dideoxy-3'-enan-kanamycin were obtained in 90% yield starting from 50B.

Example 6

1. Preparation of 3 ', 4'-O-Epoxy-penta-N-tert-butoxycarbonyl4 ", 6" -O-cyclohexylidene-2 "-O-benzoyl-kanamycin B Dissolve in 60 ml of dry pyridine 2.0 g 3 ', 4'-β-Epoxy-penta-N-tert-butoxycarbonyl4 ", 6" -O-cyclohexylidenecannamycin B, 0.8 ml of benzoyl chloride was added to the solution under ice-cooling and the mixture was stirred for half an hour for 65 hours. The reaction mixture was maintained at 5 ° C. Water (2 ml) was added and the mixture was concentrated and the resulting concentrate was poured into water (20 ml). The precipitate was filtered off and dried, yielding 2.16 g (98.3%).

2. The 4'-deoxy-4'-iodo-penta-N-tert-butoxy-

Preparation of Carbonyl-4 ", 6" -O-Cyclohexylidene-2 "-O-Benzoyl-Kanamycin B Dissolve 2.1 g of 3 ', 4'-O-epoxy-penta-N-tert-butoxycarboxylic acid in ml of acetone. onyl-4 ", 6" -O-cyclohexylidene-2 "-O-benzoyl-kanamycin B, 1.4 g of sodium iodide, 80 mg of sodium acetate and 1.4 ml of acetic acid are added, followed by ca. The mixture is refluxed for 8 hours.

After completion of the reaction, the reaction mixture is cooled, whereupon the product crystallizes and is isolated by filtration. Yield: 2.25 g (94%), m.p. 171-176 ° C (dec.).

Analysis for C 4 H ₃ N _{76 s 9} Oi formula I.

date: C = 52.62% H = 6.94%, N = 5.48% I = 9.93% found: C = 52.81% H = 7.11% N = 5.31% I = 10.39%.

3. Preparation of 3 ', 4'-dideoxy-3'-ene-penta-N-tert-butoxycarbonyl4 ", 6" -O-cyclohexylidene-2 "-O-benzoyl-kanamycin B dissolved in dry pyridine 452 mg of 4'-deoxy-4'-iodopentane-N-tert-butoxycarbonyl4 ", 6" -O-cyclohexylidene-2 "-O-benzoylcannamycin B, at 0 ° C to 5 ° C 305 mg of benzylsulfonyl chloride are added and the mixture is maintained at this temperature for half an hour.

Upon completion of the reaction, 0.18 mL of methanol was added and the mixture was heated to 90 ° C for 50 minutes. After cooling to room temperature, the solution is concentrated and the product is precipitated from the syrup obtained by adding 10 ml of water. Filter and wash the crystalline product with water.

Thin layer chromatography (silica gel, eluent mixture: carbon tetrachloride: acetone = 4: 1, v / v) identifies the product thus obtained.

4. Preparation of 3 ', 4'-dideoxy-3'-ene-penta-N-tert-butoxycarbonyl-4', 6 '-O-cyclohexylidenecannamycin B in methanol was dissolved in the wet wet substance from step 3, the solution was adjusted to pH 9.0-10.0 with methylate, stirred at room temperature for half an hour, then neutralized with 1 N hydrochloric acid and concentrated. Water is added to precipitate the product from the residual syrup, filtered, washed with water and dried. Yield: 360 mg (100%).

Claims (11)

A process for the preparation of 3 ', 4'-dideoxy-kanamycin B and its acid addition salts comprising: (a) one of the following general formulas: R is hydrogen or (C1-C6) alkyl, and Z is cyclohexylidene and the 3 ', 4'-epoxy can be in the a- or β-position - the 3', 4'-epoxy-cananicin B derivative having protected amino and hydroxy groups is treated with xanthate, so that III and IV wherein R and Z are the B 'derivative 3', 4'-dideoxy-3'-en-kanamycin and the B derivative 3 ', 4'-epithiocanamycin, respectively, wherein 3', 4 ' The '-epithio group is in the a or β position, whereupon the 3', 4'-deoxy-3'-enamen-kanamycin B derivatives of formula III, the 3'4'-dideoxy of formula III are recovered from the reaction mixture. The 3'-en-kanamycin B derivatives are cleaved in a manner known per se by the amino-protecting groups -COOR and the hydroxyl protecting groups Z, and the resulting 3 ', 4'-dideoxy-3'-en-kanamycin B of formula V is known per se. hydrogenated, or b) treating a 3 ', 4'-epoxycannamycin B derivative of the Formula II wherein R and Z are as defined above with protected amino and hydroxy groups, and thereby treating a compound of Formula IV wherein R and Z are as defined above, and the 3 ', 4'-epithio group may be in the α or β position - for the 3', 4'-epithiocannamycin B derivatives and in the formula III wherein R and Z are as defined 3 ', 4'-dideoxy-3'-enan-kanamycin B is obtained, followed by recovery of the 3', 4'-epithiocanamycin B-derivative of formula IV from the reaction mixture, followed by 3 ', 4' treating the '-epithio derivative with an acid, and hydrogenating the resulting 3', 4'-dideoxy-3'-enamamycin B of formula V in a manner known per se, or c) treating a 3 ', 4'-epoxycannamycin B derivative of the Formula II wherein R and Z have the above protected amino and hydroxy groups with a xanthate to form a compound of Formula IV wherein R and Z are the aforesaid and the 3 ', 4'-epithio group may be in the? or? - position for the 3', 4'-epithiocannamycin B derivatives, and the formula III wherein R and Z are as defined above for the 3 ', 4'-dideoxy-3'-enan-kanamycin B derivatives, and the resulting reaction mixture is treated with hydrazine or Raney nickel to give the resulting compound of formula III wherein R and Z are the amino-protecting groups -COOR of the above 3 ', 4'-dideoxy-3'-ene derivatives and the hydroxyl protecting groups of Z are removed by methods known per se, and the resulting 3', 4'-dideoxy-3'-canamycin B hydrogenation according to methods known per se and, if desired, converting the resulting 3 ', 4'-dideoxy-kanamycin B into an acid addition salt thereof. 2. The process of claim 1, wherein the amino and hydroxyl protecting groups are removed after the hydrogenation step. 3. The process of claim 1, wherein the xanthate is an alkali metal xanthate. 4. The process of claim 1, wherein the xanthate is reacted in an organic solvent. 5. A process according to claim 4, wherein the solvent is a lower alkanol. 6. A process according to claim 1, wherein the reaction with xanthate is carried out at a temperature between 50 ° C and 100 ° C. 7. The process of claim 1, wherein the chromatographic method is used to recover the 3 ', 4'-dideoxy-3'-en-kanamycin B derivatives from the reaction mixture. 8. The process of process variant b) of claim 1, wherein the acid treatment of the 3 ', 4'-epithioic derivative is carried out in a lower alkanol solvent at a temperature between 0 ° C and 30 ° C. 9. A process according to claim 8 wherein the acid is a non-oxidizing mineral acid. 10. A process according to claim 9 wherein the non-oxidizing mineral acid is a hydrohalic acid. 11. The process of claim 1, wherein the 3 ', 4'-epithio derivatives are treated with hydrazine or Raney nickel as a solvent in a lower alkanol at room temperature.