JPS632267B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention is based on the general formula (In the formula, R ₁ and R ₂ may be the same or different, each represents a hydrogen atom or a methyl group, R ₃ represents a hydrogen atom or an acyl group, and the amino group and/or hydroxyl group may be protected) The present invention relates to novel aminoglycosides and acid addition salts thereof. The acyl group R ₃ is preferably one derived from an amino acid such as glycine, alanine, or valine, and the amino group of the amino acid may be substituted with, for example, a lower alkyl group, a carbamoyl group, a formyl group, or the like. The present inventors previously reported that Saccharopolyspora hirsuta strain KC-6606 and Streptomyces sp.
We found several compounds (KA-6606 substance group and KA-7038 substance group) that are useful as antibiotics as metabolic products of strain KC-7038. These compounds are disclosed in JP-A-53-127401 and JP-A-54-127401.
No. 66603, No. 54-141701, No. 55-11541, No. 55
-111497 and No. 55-162795, and these antibiotics are represented by the following structural formula.

[Table] KA-6606, KA-7038 and KA-7038 are ranked first, respectively.
It is a stereoisomer at the 4th or 5th position. The present inventors synthesized various derivatives of these compounds and examined their activities, and found that the 5-day-O-methyl compound, which is obtained by cleaving the methyl ether at the 5-position, has excellent antibacterial activity ( Tokukai Akira
55-55198 and 56-2997). As a result of further research, the present inventors found that the above-mentioned 5-day
A compound in which the hydroxyl group at the 5-position of the O-methyl compound has been removed,
That is, the present invention was achieved by discovering that 5-de-methoxy compound exhibits even more excellent antibacterial action. The compound of formula (1) is, for example, a compound of the general formula It is produced by eliminating (or deoxylating) the hydroxyl group at the 5-position of a compound represented by the formula (wherein R ₁ , R ₂ and R ₃ have the meanings described above). Before this deoxylation, each amino group and the hydroxyl group at the 3-position of the compound of formula (3) are preferably protected by a conventional method. The compound of formula (3) has the general formula (In the formula, R ₁ , R ₂ and R ₃ have the above meanings) (KA-6606 substance and KA
-7038 substance) by the method described in JP-A-55-55198, mineral acids such as hydrobromic acid, hydroiodic acid, hydrofluoric acid, etc. or Lewis acids such as boron trichloride, trifluoride. It can be produced by cleaving the 5-position methyl ether under the action of boron, etc., or an alkali metal and an amine. Prior to cleavage of the methyl ether (de-O-methylation) each amino and/or hydroxyl group is preferably protected. When a compound of formula (4) in which R ₃ is an acyl group is used, the acyl group R _{3 and the optionally present amino protecting group are also removed at the same time as de-O-methylation, and the compound of the formula (4} ) in which R ₃ is a hydrogen atom is removed. Compound (3) is obtained. Then, if desired, an acyl group R ₃ can be introduced. Various methods can be applied to deoxylate the 5-position of the compound of formula (3), and for example, the following method is preferred. (A) Sulfamoylation of the hydroxyl group at position 5, for example N.
After N-dimethylsulfamoylation or sulfonylation, such as trifluoromethanesulfonylation, the protecting group of the amino group is removed or not removed, so the carbon-oxygen bond is cleaved, for example, by reaction with metallic sodium in liquid ammonia. Method. (B) A method in which the 5-position hydroxyl group is substituted with a halogen atom to form a 5-halo (preferably 5-chloro) compound, and then, for example, tri-n-butylstannane is applied to dehalogenate the compound. (C) A method in which the 5-position hydroxyl group is esterified with phosphoric acid, preferably a substituted phosphoric acid such as bis-dimethylaminophosphoric acid, and then reduced with lithium in the presence of, for example, ethylamine. (D) A method in which the hydroxyl group at the 5-position is carbonylated to form a hydrazone, such as tosylhydrazone, and then reduced with, for example, sodium cyanoborohydride. (E) Thiobenzoate, S-methyldithiocarbonate, etc. are formed at the 5-position, and tri-
A method of making n-butylstannane act. (F) A method in which a sulfonic acid ester is prepared, followed by halogen substitution (preferably iodine substitution), and then catalytic reduction. (G) After trifluoromethanesulfonylation,
How to photolyse. Before performing these reactions for deoxylating the hydroxyl group at the 5-position, each amino group of compound (3) and 3
It is preferable to protect the hydroxyl group at this position. As the protecting group for the amino group, known amino protecting groups commonly used in conventional peptide synthesis can be used, but the general formula A protecting group represented by (wherein A represents an alkyl group, a cycloalkyl group, an aryl group, or an aralkyl group) is particularly preferred. Examples of these protecting groups include the following. Alkyloxycarbonyl groups such as ethyloxycarbonyl group, tertiary butyloxycarbonyl group, tertiary amyloxycarbonyl group, etc., cycloalkyloxycarbonyl group such as cyclohexyloxycarbonyl group, aryloxycarbonyl group such as phenoxycarbonyl group, etc. Alkyloxycarbonyl group such as benzyloxycarbonyl group, p
-methoxybenzyloxycarbonyl group, etc. The introduction of a protecting group is carried out by a conventional method, but a carboxylic acid represented by the general formula A-OCOOH (5) (A in the formula has the meaning described above) or a reactive derivative thereof is applied to compound (3). The preferred method is to Examples of reactive derivatives of carboxylic acids include acid halides, acid azide compounds, acid anhydrides, and active esters. Then, only the hydroxyl group at position 3 is selectively protected. The compound of formula (3) has 4 adjacent to the 3-position hydroxyl group.
Since there is an amino group at the position, by utilizing this to form a cyclic carbamate, the hydroxyl group at the 3-position can be easily protected together with the amino group at the 4-position. Cyclic carbamates are formed by the action of a tetra-N-protected compound with an alkali, preferably in a solvent. Examples of alkali include sodium hydride, barium hydroxide,
For example, dimethylformamide, dioxane, tetrahydrofuran, water, etc. or a mixture thereof can be used as the solvent, such as sodium hydroxide or calcium hydroxide. When the methylamino group at the 4-position of the compound of formula (1) in which R ₃ is a hydrogen atom is optionally acylated, R ₃
A compound of formula (1) is obtained in which is an acyl group. To carry out the acylation, the methylamino group at the 4-position is first released and then acylated. That is, by hydrolyzing the cyclic carbamate between the 3rd and 4th positions, the methylamino group at the 4th position can be easily released. For this hydrolysis, ordinary acid hydrolysis or alkaline hydrolysis can be applied. If the protecting group is also removed at the same time during the deoxylation reaction, the amino groups at the 1-, 2'- and 6'-positions of the compound of formula (1) are first protected. This protection reaction can be carried out in the same manner as the above-mentioned protection reaction of the amino group of compound (3). At this time, since the 4-position methylamino group is also protected, it is necessary to liberate the 4-position methylamino group to be acylated. For this purpose, by forming a cyclic carbamate between the hydroxyl group at the 3-position and the methylamino group at the 4-position in the same manner as described above, and then hydrolyzing this, it is easy to remove only the methylamino group at the 4-position. It can be liberated. For hydrolysis, ordinary acid hydrolysis or alkaline hydrolysis can be applied. Furthermore, when a tetra-N-protected compound is treated with an alkali in a solvent containing water, only the methylamino group at the 4-position can be directly released. On the other hand, when protecting the amino group of the compound of formula (3) or (1) where R ₃ =H, active esters such as substituted phenyl esters, N- Oxysuccinimide ester, N-oxyphthalimide ester, etc., especially general formula (In the formula, X represents an optionally substituted phenyl group, succinimidyl group, or phthalimidyl group)
When the active ester represented by is used, the 1st position,
Only the amino groups at the 2' and 6' positions can be selectively protected. Favorable results are obtained when a metal compound such as nickel acetate, cobalt acetate, copper acetate, etc. is present during this reaction. Compounds of formula (1) with protected amino groups at positions 1, 2' and 6' obtained by these methods (R ₃ = H)
When the methylamino group at the 4-position is acylated, the compound of formula (1) with a protected amino group (R ₃ = acyl)
is obtained. As the acylating agent, it is preferable to use amino acids such as glycine, alanine, valine, and the like. For acylation using amino acids, normal peptide synthesis methods can be applied, and N-
A protected amino acid or a reactive derivative thereof is applied. As the protecting group for the amino group of the amino acid, any of the above-mentioned protecting groups may be used, and it may be the same as or different from the protecting group of the compound of formula (1) (R ₃ = H), but in the next step It is preferable to use the same protecting group in order to easily carry out the elimination reaction in . In addition, the amino group may be substituted with, for example, a lower alkyl group, a carbamoyl group, a formyl group, or the like.
As the acid-reactive derivative of the amino acid, those similar to the carboxylic acid-reactive derivative of formula (5) can be used. The thus obtained compound of formula (1) with a protected amino group and/or hydroxyl group (R ₃ = H or acyl group)
Removal of the protecting group gives the free compound of formula (1). Although conventional methods can be applied to remove the protecting groups for amino groups and hydroxyl groups, it is preferable to use catalytic reduction and acid hydrolysis. Examples of catalysts used for catalytic reduction include palladium, platinum, Raney nickel, rhodium, ruthenium, and nickel. As the solvent, for example, water, methanol, ethanol, dioxane, tetrahydrofuran, acetic acid, etc. or a mixture thereof can be used. This reaction is usually completed within 0.1 to 10 hours at a hydrogen pressure of 1 to 5 atmospheres and a temperature of 0 to 100°C. Isolation and purification of the target substance, the compound of formula (1), and the compound with its amino group and/or hydroxyl group protected, can be carried out by conventional methods, but it is particularly preferable to use column chromatography. As the adsorbent, it is preferable to use a cation exchange resin such as CM-Sephadex, Amberlide IRC-50, Amberlide IRC-84, Amberlide CG-50, and carboxymethyl cellulose. The development can be carried out by a concentration gradient method or a concentration step method using an alkaline aqueous solution such as aqueous ammonia or aqueous ammonium chloride solution as a developing solvent. By collecting the active fraction from the eluate and lyophilizing it, a pure product of the target compound can be obtained. The compound of formula (1) can be converted into an acid addition salt according to a conventional method. As acids, inorganic acids such as sulfuric acid, hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, carbonic acid, nitric acid, etc. and organic acids such as acetic acid, fumaric acid, malic acid, citric acid, mandelic acid, succinic acid, etc. are used. It will be done. Representative examples of the new compounds of formula (1) include 5-demethoxy-KA-6606 (A), 4-N-glycyl-5
The antibacterial spectra of -demethoxy-KA-6606 (B) and 5-demethoxy-KA-7038 (C) are compared to KA-6606, which has a methoxy group at the 5-position, respectively.
(A'), 4-N-glycyl-KA-6606 (B') and KA-7038 (C') are shown in the following table.

【table】

[Table] Example 1 (a) The following formula The compound represented by (5-de-O-methyl-
Dissolve 1 g of KA-6606) in 10 ml of water, add 1.3 g of anhydrous sodium carbonate and 40 ml of methanol, and then drop 2.6 ml of carbobenzyloxychloride under ice-cooling. After continuing stirring for an additional 3 hours under ice-cooling, the reaction solution was concentrated to dryness. Chloroform was added to the residue, washed with water, dried, and the solvent was distilled off to obtain 2.6 g of colorless crystals. When this crystal is recrystallized from benzene, the melting point is 153-154.
1,4,2',6'-tetrakis-N-benzyloxycarbonyl-5-de-O-methyl-KA-6606 is obtained as colorless needle-like crystals at . Elemental analysis value: C ₄₆ H ₅₄ N ₄ O ₁₂ Calculated value (%) 64.62 6.37 6.55 Actual value (%) 64.49 6.33 6.61 Specific rotation: [α] ²³ _D +44° (c1, CHCl ₃ ) ( b) Dissolve 100 mg of the N-protected compound obtained in (a) in 2 ml of dioxane, add 1.5 ml of 0.1M barium hydroxide aqueous solution, and stir at 60°C for 1 hour.
Add dry ice to the reaction solution to neutralize it, filter out insoluble matter, and concentrate the solution to dryness. The residue was separated by preparative thin layer chromatography (chloroform-methanol 15:1) to give 1,2',6'-tris-N-benzyloxycarbonyl-3-O:4-N-carbonyl- as a colorless solid. 668 mg of 5-deO-methyl-KA-660 is obtained. Elemental analysis value: C ₃₉ H ₄₆ N ₄ O ₁₁ C H N Calculated value (%) 62.72 6.21 7.50 Actual value (%) 62.48 6.10 7.28 Specific rotation: [α] ²³ _D +33° (c1, CHCl ₃ ) IR Value: ν ^CHCl3 _nax cm ^-1 1760 (cyclic carbamate) PMR value: δ _CDCl3 1.07 (3H, d, J=6Hz, C-C H ₃ ) 2.87 (3H, s, N-C H ₃ ) (c) ( 223 mg of N.O-protected compound obtained by b)
Dissolve in 5 ml of anhydrous dimethylformamide and add 50
Add 43mg of % oily sodium hydride under ice cooling,
Next, 0.12 ml of N.N-dimethylsulfamoyl chloride was added, and the mixture was stirred for 1 hour under ice cooling. After adding 0.4 ml of acetic acid to the reaction solution, 50 ml of ice-cold water was added, and the precipitate was collected, washed with water and n-hexane, and then dried. The resulting colorless powder was purified by preparative thin layer chromatography (chloroform-methanol 20:1) to yield 1,2',6'-tris-N-benzyloxycarbonyl-3-O:4- as a colorless solid. N-carbonyl-5-deO-methyl-5-O-(N.
N-dimethylsulfamoyl)-KA-6606
167mg is obtained. Elemental analysis value: C ₄₁ H ₅₂ N ₅ O ₁₃ S Calculated value (%) 57.60 6.13 8.19 3.75 Actual value (%) 57.86 6.01 7.93 3.44 Specific optical rotation: [α] ²³ _D +35° (c1, _CHCl3 ) IR value: ν ^CHCl3 _nax cm ^-1 1765 (cyclic carbamate) 1170 (sulfamoyl) PMR value: δ _CDCl3 1.08 (3H, d, J=6.5Hz, C-C H ₃ ) 2.84 (6H, s, SO _2N (C H ₃ ) ₂ ) 2.92(3H, s, N-C H ₃ ) (d) 135 mg of the 5-O-dimethylsulfamoyl compound obtained in (c) was dissolved in 3 ml of acetic acid, and used as a catalyst. Catalytic reduction is carried out using palladium black at room temperature and pressure. The catalyst was removed from the reaction solution, and the solution was concentrated to dryness. Dissolve the residue in 100ml of water,
It was adsorbed on a 5 ml column of CM-Sephadex C-25 (NH ₄ ⁺ type), washed with water, eluted with 0.7M aqueous ammonia, and the eluate was concentrated to dryness. Dissolve the residue in 3 ml of liquid ammonia, add 100 mg of metallic sodium at -70°C, and stir for 3 hours. Water is added to the reaction solution to decompose sodium, and then concentrated to dryness. The residue was dissolved in 2 ml of water, heated at 60°C for 3 hours, diluted to 100 ml with water, and neutralized with 4N sulfuric acid. This is passed through a CM-Sephadex column and treated in a conventional manner, resulting in a colorless solid, 5-demethoxy-KA-
660615mg is obtained. Elemental analysis value: C ₁₄ H ₃₀ N ₄ O ₃・H ₂ O Calculated value (%) 52.48 10.07 17.49 Actual value (%) 52.31 9.83 17.24 Specific rotation: [α] ²² _D +119° (c1, H ₂ O) PMR value: δ _D2O (TMS external standard) 1.51 (3H, d, J = 7Hz, C-C H ₃ ) 2.80 (3H, s, N-C H ₃ ) 5.44 (1H, d, J = 3.7Hz, anomeric H) 1.7-2.4 (8H, m, C- _CH2 -C) Example 2 (a) 5-demethoxy-KA-6606 (Example 1d)
Dissolve 30mg in 1ml of methanol, add 75mg of nickel acetate (tetrahydrate) and stir for 30 minutes.
Add 80 mg of benzyloxycarbonyloxysuccinimide and stir at room temperature for 3 hours. Add 1 ml of concentrated ammonia water to the reaction solution, stir for 1 hour, and then concentrate to dryness. Chloroform and residue
Add 10ml each of 4N ammonia water, separate the chloroform layer, wash with 4N ammonia water,
Dry and remove the solvent. Dioxane the residue
Dissolve in 2.3 ml of triethylamine and 0.15 ml of N-benzyloxycarbonylglycine.
Add 80 mg of hydroxysuccinimide ester and warm at 60°C overnight. The reaction solution was concentrated, and the residue was subjected to preparative thin layer chromatography (silica gel, chloroform-methanol 13:
When separated and purified in step 1), colorless solids 1, 2',
6'-Tris-N-benzyloxycarbonyl-
661 mg of 4-N-(N'-benzyloxycarbonylglycyl)-5-demethoxy-KA-6606, ie tetrakis-N-benzyloxycarbonyl-5-demethoxy-KA-660, is obtained. Elemental analysis value: C ₄₈ H ₅₇ N ₅ O ₁₂ C H N Calculated value (%) 64.34 6.41 7.82 Actual value (%) 64.51 6.43 7.56 Specific rotation: [α] ²³ _D +38° (c2, CHCl ₃ ) IR Value: ν ^CHCl3 _nax cm ^-1 1635 (amide) PMR value: δ _CDCl3 1.08 (3H, d, J=6.5Hz, C-C H ₃ ) 2.88 (3H, s, N-C H ₃ ) (b) Obtained 60 mg of the obtained N-protected compound was dissolved in 1 ml of acetic acid and subjected to catalytic ring reduction using palladium black as a catalyst. After the reaction is complete, remove the catalyst, dilute to 160 ml with water, and neutralize with aqueous ammonia. This solution
The target substance is adsorbed through a 12 ml column of CM-Sephadex C-25 (NH ₄ ⁺ type), washed with water, and then eluted between 0.05N and 0.5N aqueous ammonia using the concentration gradient method. When the part containing the target substance is freeze-dried, 621 mg of 5-demethoxy-KA-660, which is a colorless solid and is represented by the following formula, is obtained. Elemental analysis value: C ₁₆ H ₃₃ N ₅ O ₄・H ₂ O Calculated value (%) 50.91 9.35 18.55 Actual value (%) 50.68 9.21 18.23 Specific optical rotation: [α] ²² _D +118° (c1, H ₂ O) PMR value: δ _D2O 1.51 (3H, d, J=6.5Hz, C-C H ₃ ) 1.8-2.4 (8H, m, C-C H ₂ -C) 3.45 (3H, s, N- C H ₃ ) 3.99 (2H, s,

[Formula] ) 5.45 (1H, d, J=4Hz, anomeric H) Example 3 (a) The following formula The compound represented by (5-de-O-methyl-
When 300 mg of KA-6606) was used and treated in the same manner as in Example 1(a), a colorless solid of tetrakis-
N-benzyloxycarbonyl-5-deO-
6,760 mg of methyl-KA-660 is obtained. Elemental analysis value: C ₄₆ H ₅₄ N ₄ O ₁₂ Calculated value (%) 64.62 6.37 6.55 Actual value (%) 64.90 6.18 6.48 Specific rotation: [α] ²² _D +51° (c1, CHCl ₃ ) PMR Value: δ _CDCl3 1.16 (3H, d, J=6Hz, C-C H ₃ ) 2.89 (3H, s, N-C H ₃ ) 7.3-7.4 (20H, m, 4×C ₆ H ₅ ) (b) Example 1 using 540 mg of the above N-protected compound
Operate in the same way as (b). Silica gel column chromatography (chloroform-methanol 20:
When separated and purified in step 1), colorless solids 1, 2',
6'-Tris-N-benzyloxycarbonyl-
3-O: 4-N-carbonyl-5-de-O-methyl-KA-6606384 mg is obtained. Elemental analysis value: C ₃₉ H ₄₆ N ₄ O ₁₁ C H N Calculated value (%) 62.72 6.21 7.50 Actual value (%) 62.70 5.90 7.34 Specific rotation: [α] ²² _D +48° (c1, CHCl ₃ ) IR Value: ν ^CHCl3 _nax cm ^-1 1760 (cyclic carbamate) PMR value: δ _CDCl3 1.15 (3H, d, J=6Hz, C-C H ₃ ) 2.87 (3H, s, N-C H ₃ ) (c) Above Using 318 mg of the N.O-protected compound, the same procedure as in Example 1(c) resulted in a colorless solid of
2',6'-tris-N-benzyloxycarbonyl-3-O: 4-N-carbonyl-5-deO
-Methyl-5-O-(N.N-dimethylsulfamoyl)-KA-6606275 mg is obtained. Elemental analysis value: C ₄₁ H ₅₂ N ₅ O ₁₃ S Calculated value (%) 57.60 6.13 8.19 3.75 Actual value (%) 57.51 5.87 7.98 3.63 Specific optical rotation: [α] ²² _D +39° (c1, _CHCl3 ) IR value: ν ^CHCl3 _nax cm ^-1 1760 (cyclic carbameth), 1170 [SO ₂ N
(CH ₃ ) ₂ ] PMR value: δ _CDCl3 1.17 (3H, d, J=6Hz, C- CH ₃ ) 2.85 [6H, s, SO ₂ N (CH ₃ ) ₂ ] 2.89 (3H, s, N -C H ₃ ) (d) Using 260 mg of the above-mentioned 5-O-dimethylsulfamoyl compound, the same procedure as in Example 1(d) is carried out.
When purified by CM-Sephadex C-25 (NH ₄ ⁺ form) ion-exchange chromatography, a colorless solid, 5-de-methoxy-KA, represented by the following formula
-660638 mg is obtained. Elemental analysis value: C ₁₅ H ₃₀ N ₄ O ₃・1/2H ₂ CO ₃・H ₂ O
As C H N Calculated value (%) 49.55 9.46 15.94 Actual value (%) 49.25 9.18 15.81 Specific rotation: [α] ²² _D +72° (c1, H ₂ O) PMR value: δ _D2O (TMS external standard) 1.53 ( 3H, d, J = 6.5Hz, C-C H ₃ ) 2.86 (3H, s, N-C H ₃ ) 5.52 (1H, d, J = 3.5Hz, Anomeric H) 1.7-2.4 (8H, m, CC _H2 -C) Example 4 (a) 5-demethoxy-KA-6606 (Example 3(d)
Using 26 mg and operating in the same manner as in Example 2(a),
Colorless solid 1,2',6'-tris-N-benzyloxycarbonyl-4-N-(N'-benzyloxycarbonylglycyl)-5-demethoxy-
KA-660658 mg is obtained. Elemental analysis value: C ₄₈ H ₅₇ N ₅ O ₁₂ C H N Calculated value (%) 64.34 6.41 7.82 Actual value (%) 64.21 6.19 7.62 Specific rotation: [α] ²² _D +28° (c1, CHCl ₃ ) IR Value: ν ^CHCl3 _nax cm ^-1 1640 (amide) PMR value: δ _CDCl3 1.19 (3H, d, J=6Hz, C-C H ₃ ) 2.90 (3H, s, N-C H ₃ ) (b) Above Example 2 using 48 mg of N-protected compound
By operating in the same manner as in (b), a colorless solid, 5-demethoxy-4-N-glycyl-
KA-660614 mg is obtained. Elemental analysis value: C ₁₆ H ₃₃ N ₅ O ₄・3/2H ₂ O Calculated value (%) 49.72 9.39 18.12 Actual value (%) 49.97 9.08 18.03 Specific rotation: [α] ²² _D +108° ( c1, H ₂ O) PMR: δ _D2O (TMS external standard) 1.53 (3H, d, J=6.5Hz, C-C H ₃ ) 3.58 (3H, s, N-C H ₃ ) 4.01 (2H, s, CO-C H ₂ -NH ₂ ) 5.40 (1H, d, J=3.3Hz, anomeric H) Example 5 (a) The following formula The compound represented by (5-de-O-methyl-KA
-7038) Using 330 mg, the same procedure as in Example 1(a) yielded a colorless solid, tetrakis-N-benzyloxycarbonyl-5-de-O-methyl-KA-
7038820 mg is obtained. Elemental analysis value: C ₄₆ H ₅₄ N ₄ O ₁₂ Calculated value (%) 64.62 6.37 6.55 Actual value (%) 64.59 6.21 6.38 Specific rotation: [α] ²² _D +62° (c1, CHCl ₃ ) PMR Value: δ _CDCl3 2.89 (3H, s, N-C H ₃ ) 2.92 (3H, s, N-C H ₃ ) (b) Same as Example 1(b) using 795 mg of the above N-protected compound. When manipulated, the colorless solid 1, 2',
6'-Tris-N-benzyloxycarbonyl-
3-O: 503 mg of 4-N-carbonyl-5-deO-methyl-KA-7038 is obtained. Elemental analysis value: C ₃₉ H ₄₆ N ₄ O ₁₁ C H N Calculated value (%) 62.72 6.21 7.50 Actual value (%) 62.45 6.02 7.28 Specific rotation: [α] ²² _D +46° (c1, CHCl ₃ ) IR Value: ν ^CHCl3 _nax cm ^-1 1760 (cyclic carbamate) PMR value: δ _CDCl3 2.81 (3H, s, N-C H ₃ ) 2.90 (3H, s, N-C H ₃ ) (c) The above N・O - Using 480 mg of the protected compound and operating in the same manner as in Example 1(c), a colorless solid of 1.
2',6'-tris-N-benzyloxycarbonyl-3-O: 4-N-carbonyl-5-deO
-Methyl-5-O-(N.N-dimethylsulfamoyl)-KA-7038411 mg is obtained. Elemental analysis value: C ₄₁ H ₅₂ N ₅ O ₁₃ S Calculated value (%) 57.60 6.13 8.19 3.75 Actual value (%) 57.50 5.88 7.86 3.71 Specific optical rotation: [α] ²² _D +41° (c1, _CHCl3 ) IR value: ν ^CHCl3 _nax cm ^-1 1760 (cyclic carbamate) 1170 (sulfamoyl) PMR value: δ _CDCl3 2.82 (3H, s, N-C H ₃ ) 2.91 (3H, s, N-C H ₃ ) (d) 300 mg of the above 5-O-sulfamoyl compound
By using 5-demethoxy-KA represented by the following formula as a colorless solid and operating in the same manner as in Example 1(d),
−703831 mg is obtained. Elemental analysis value: C ₁₄ H ₃₀ N ₄ O ₃・H ₂ O Calculated value (%) 52.48 10.07 17.49 Actual value (%) 52.24 9.99 17.23 Specific rotation: [α] ²² _D +36° (c1, H ₂ O) PMR value: δ _D2O (TMS external standard) 2.80 (3H, s, N-C H ₃ ) 2.83 (3H, s, N-C H ₃ ) 5.54 (1H, d, J=3.3Hz, anomeric H) Example 6 (a) 5-demethoxy-KA-7038 (Example 5d)
Using 26 mg and operating in the same manner as in Example 2(a),
Colorless solid 1,2',6'-tris-N-benzyloxycarbonyl-4-N-(N'-benzyloxycarbonylglycyl)-5-demethoxy-
KA-7038, i.e., tetrakis-N-benzyloxycarbonyl-5-demethoxy-KA
−703846 mg is obtained. Elemental analysis value: C ₄₈ H ₅₇ N ₅ O ₁₂ Calculated value (%) 64.34 6.41 7.82 Actual value (%) 64.08 6.20 7.96 Specific rotation: [α] ²² _D +32° (c1, CHCl ₃ ) PMR Value: δ _CDCl3 2.91 (6H, s, N-C H ₃ ) (b) Using 40 mg of the above N-protected compound, Example 2
By operating in the same manner as in (b), 12 mg of 5-demethoxy-KA-703, represented by the following formula, as a colorless solid is obtained. Elemental analysis value: C ₁₆ H ₃₃ N ₅ O ₄・H ₂ CO ₃ Calculated value (%) 48.44 8.37 16.62 Actual value (%) 48.21 8.02 16.74 Specific rotation: [α] ²² _D +112° (c1 , H ₂ O) PMR value: δ _D2O (TMS external standard) 2.84 (3H, s, 6'-N-C H ₃ ) 3.61 (3H, s, 4-N-C H ₃ ) 5.40 (1H, d, J=3Hz, anomeric H) Example 7 (a) 1・2′・6′-tris- obtained in Example 1(b)
N-benzyloxycarbonyl-3-O:4-
N-carbonyl-5-deO-methyl-KA-
Dissolve 66062g in 40ml of dichloromethane, -
4 ml of pyridine and sulfuryl chloride at 10°C.
Add 1.6 ml and incubate at -10℃ for 12 hours, then at 5℃ for 4 hours.
Let stand for a while. The reaction solution was poured into a mixture of 200 ml each of chloroform and saturated sodium bicarbonate solution. After separating the chloroform layer and washing it with saturated sodium bicarbonate solution,
Dry and remove the solvent. Add 40ml of benzene to the residue
The solution was dissolved in water, heated at 70°C for 3 hours, and the solvent was distilled off. The residue was subjected to silica gel column chromatography and chloroform-methanol (100:
1), a colorless solid of 1,2',6'-tris-N-benzyloxycarbonyl-3-
66061.3 g of O:4-N-carbonyl-5-chloro-5-demethoxy-KA-660 is obtained. Elemental analysis value: C ₃₉ H ₄₅ ClN ₄ O ₁₀ C H N Cl Calculated value (%) 61.21 5.93 7.32 4.63 Actual value (%) 60.98 5.81 7.51 4.55 Specific optical rotation: [α] ²³ _D +24° (c2, CHCl ₃ ) IR value: ν ^CHCl3 _nax cm ^-1 1760 (cyclic carbamate) PMR value: δ _CDCl3 1.06 (3H, d, J=6Hz, C-C H ₃ ) 2.86 (3H, s, N-C H ₃ ) ( b) 1.2 g of the above 5-chloro compound was added to 24 g of toluene.
1.5 ml of tri-n-butylstannane
After replacing the mixture with nitrogen, add 20 mg of azobisisobutyronitrile and heat at 80°C for 4 hours.
The reaction solution was concentrated to dryness, the residue was applied to a silica gel column, and chloroform-methanol (100:
1), a colorless solid of 1,2',6'-tris-N-benzyloxycarbonyl-3-
O: 4-N-carbonyl-5-demethoxy-
KA-66061.13g is obtained. Elemental analysis value: C ₃₉ H ₄₆ N ₄ O ₁₀ C H N Calculated value (%) 64.10 6.34 7.67 Actual value (%) 63.95 6.32 7.61 Specific rotation: [α] ²⁴ _D +3° (c2, CHCl ₃ ) IR Value: ν ^CHCl3 _nax cm ^-1 1755 (cyclic carbamate) PMR value: δ _CDCl3 1.07 (3H, d, J=7Hz, C-C H ₃ ) 2.73 (3H, s, N-C H ₃ ) (c) Above 1.1g of 5-demethoxy form of dioxane
Dissolve in 20ml 0.3M barium hydroxide solution 25
ml and stirred at 60°C overnight. The reaction solution is neutralized with carbon dioxide gas to remove insoluble matter, and the solution is concentrated to dryness. Dissolve the residue in 10 ml of acetonitrile and add N-benzyloxycarbonylglycine.
500mg and dicyclohexylcarbodiimide 500
After stirring at room temperature for 3 hours, insoluble matter was removed and the liquid was concentrated to dryness. The residue was eluted with chloroform-acetone (6:1) on a silica gel column to obtain 820 mg of tetrakis-N-benzyloxycarbonyl-5-demethoxy-KA-6606 as a colorless solid. The specific rotation, IR value, and PMR value of this compound match those of the specimen obtained in Example 2(a). (d) 5-Demethoxy-KA-6606 is obtained from the above N-protected compound in the same manner as in Example 2(b).

Claims (1)

[Claims] 1. General formula (In the formula, R ₁ and R ₂ may be the same or different, each represents a hydrogen atom or a methyl group, R ₃ represents a hydrogen atom or an acyl group, and the amino group and/or hydroxyl group may be protected) The represented compound or its acid addition salt.