JPH0582394B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a method for producing 3'-fluoro-3'-deoxykanamycin A. The present inventors disclosed 3'-fluoro-3'-deoxykanamycin A as a novel semi-synthetic aminoglycoside antibiotic in the previous Japanese Patent Application No. 161615/1984 (see Japanese Patent Application Laid-open No. 40297/1982). In addition, for the production of this new compound, 3-deoxy-3-fluoro-1,2;5,
A synthetic method has been described starting from 6-di-O-isopropylidene-α-D-glucofuranose via a multistep route. Since the synthesis method of 3'-fluoro-3'-deoxykanamycin A described in the above patent application is complicated and low in efficiency, 3'-fluoro-3'-deoxykanamycin A is produced starting from kanamycin A. Further research was carried out with the intention of developing a method. As a result, starting from kanamycin A, known N,O-protected derivatives of kanamycin A (Japanese Patent Application Laid-Open No. 55-105699,
and JP-A-56-68698, or “Carbohydrate Research”
89 , pp. 91-101 (1981)) is the following formula:

[Chemical formula] [In the formula, Ts represents a tosyl group]
6′-N,4′-O-carbonyl-4″,6″-O-cyclohexylidene-1,3,3″-tri-N-tosylkanamycin A [referred to as compound (a)] was prepared. Compound (a) is reacted with a 2.5-5 molar ratio of N-acetylimidazole in anhydrous dimethyl sulfoxide in the presence of pyridine at a temperature of 0-50°C.
It has been found that only the 2'-hydroxyl group and 2''-hydroxy group of compound (a) can be selectively acetylated without acetylating the 3'-hydroxyl group.The 6'-N, 4′-O-carbonyl-4″, 6″-O-cyclohexylidene-2′,
2″-di-O-acetyl-1,3,3″-tri-N-
Tosylkanamycin A [compound (b)] is prepared by treating it with a benzylsulfonylating agent such as benzylsulfonyl chloride, a mesylating agent such as mesyl chloride, or a tosylating agent such as tosyl chloride in anhydrous pyridine at a temperature below 50°C. When you react,
The fact that the hydroxyl group of compound (b) is hardly sulfonylated and the 3'-hydroxyl group can be substantially selectively sulfonylated; therefore, 3'-O-sulfonyl-6'-N,4'-O- Carbonyl-4″,6″-O-
Cyclohexylidene-2′,2″-di-O-acetyl-1,3,3″-tri-N-tosyl-kanamycin A [referred to as compound (c)] can be produced, and the 3′ of compound (b) - The benzylsulfonylating agent is best suited for the selective sulfonylation of hydroxyl groups, and furthermore, when a mesylating agent is used, compound (b)
The 5-hydroxyl group of is partially sulfonylated, provided that the resulting 3',5-di-O-sulfonylated product is purified by chromatographic techniques to obtain the desired mono-3'-O-sulfonyl derivative, That is, it was found that it could be separated from compound (c). this compound
(c) is a reaction temperature of 0 to 50℃ and 10 minutes to 2
Under mild reaction conditions selected within the reaction time range of hours, e.g., reaction temperature at room temperature and reaction time of 10 to 20 minutes, 0.1 to
Using 0.8 molar ratio of sodium methylate, 1
Treatment with sodium methylate in anhydrous methanol containing sodium methylate at a concentration of ~5% (wt) yields 2'-O-acetyl without elimination of the 4',6'-cyclic carbamate moiety (O=C). group and the 2″-O-acetyl group can be eliminated, thus forming the 3′-
O-sulfonyl-6'-N,4'-O-carbonyl-
4″,6″-O-cyclohexylidene-1,3,3″-
It has been found that tri-N-tosylkanamycin A [referred to as compound (d)] can be produced. Furthermore, the compound
(d) is treated with sodium methylate or sodium methylate in a molar ratio of 2 to 10 to compound (d) at a reaction temperature of 0°C to the boiling point of methanol or ethanol and a reaction time of 5 minutes to 2 hours or more. sodium·
ethylate or sodium benzylate in absolute methanol containing sodium methylate at a concentration of 1 to 5% (by weight) or in absolute ethanol containing sodium methylate at a concentration of 1 to 5% (by weight) or in the presence of benzyl alcohol. When treated with 1 to 5% by weight of sodium benzylate, the 3'-sulfonyloxy group and 2'-hydroxyl group of compound (d) react to cause epoxidation, and at the same time, the compound (d) undergoes epoxidation. One of the bonds of the carbonyl group (O=C) that bridged the 6'-amino group and 4'-hydroxyl group in d) is broken from the 4'-hydroxyl group and reacts with methanol, ethanol, or benzyl alcohol. , the 6'-amino group is converted into the form of a methoxycarbonylamino group or an ethoxycarbonylamino group or a benzyloxycarbonylamino group, thus giving the formula

[Chemical formula] [In the formula, Ts is a tosyl group, and A″ is a methoxycarbonyl group, an ethoxycarbonyl group, or a benzyloxycarbonyl group] 2′,
3'-Anhydro-4'',6''-O-cyclohexylidene-3'-epi-6'-N-methoxy (or ethoxy or benzyloxy)carbonyl-1,3,3''-
It has been found that tri-N-tosylkanamycin A [referred to as compound (e)] can be produced. In addition, when reacting sodium methylate, sodium ethylate, etc. with compound (e) in anhydrous methanol or anhydrous ethanol, the reaction temperature, reaction time, and working concentration at which sodium methylate or sodium ethylate, etc. is reacted. , etc. If the reaction conditions are made more severe than the appropriate conditions mentioned above, 2'-O-
The acetyl group and the 2″-O-acetyl group are eliminated, but at the same time, the 4′,6′-cyclic carbamate moiety (O=C) from compound (c) is decomposed and the 3′- O-sulfonyl-4'',6''-O-cyclohexylidene-1,3,3''-tri-N-tosylkanamycin A [referred to as compound (d')] is produced.
This compound (d') is obtained by converting it into a compound as described above.
When treated with sodium methylate in methanol or sodium ethylate in ethanol under the same reaction conditions as when (d) is treated with sodium methylate or sodium ethylate,
2′,3′-anhydro-4″,6″-O-cyclohexylidene-3′-epi-1,3,3″-tri-N-tosylkanamycin A and 3′,4′-anhydro-4″ ，
6″-O-cyclohexylidene-3′-epi-1,
3,3″-tri-N-tosylkanamycin A is produced in a mixed state, and both the 2′,3′-epoxy form and the 3′,4′-epoxy form can be produced using conventional chromatography techniques. It was recognized that a similar hybrid epoxidation reaction could not be isolated from the other using the method.
Already shown on pages 6 and 8 of the issue. Furthermore, for compound (c), 2'-O-
With the 2′-acetoxy group of compound (c) omitted the treatment with alkali metal alcoholate for selective elimination of the acetyl group and 2″-O-acetyl group.
When compound (c) is immediately treated with an alkali metal alcoholate under reaction conditions strong enough to cause epoxidation by reaction with the 3'-sulfonyloxy group,
During the treatment, elimination of the 4',6'-cyclic carbamate moiety occurs in compound (c), and as in the case described above when compound (d') was treated, the 2',3'-epoxy and 3' ，
It is disadvantageous that the 4'-epoxy compound is produced in a mixed state. This 3',4'-epoxy compound is not directly useful for the synthesis of 3'-fluoro-3'-deoxykanamycin A, which is the objective of the method of the present invention, and is also difficult to remove chromatographically. ,3′,4′−
Epoxy by-products are undesirable. Therefore, compound (b) is produced from compound (a), and compound (b) is produced from compound (b).
(c), and following the route to produce compound (e) from compound (c), prepared compound (e) in the form of a 2',3'-epoxy-3'-epicanamycin A derivative. was recognized as necessary to provide a highly efficient method for Furthermore, as a result of repeated experiments and research, the present inventors discovered that compound (e) can be converted to sodium hydrogen difluoride by using a liquid lower alkylene glycol such as ethylene glycol or propylene glycol as a reaction medium. or with potassium hydrogen difluoride
When reacted at a temperature of 100 to 200°C for a reaction time of, for example, 1 hour to about 1 day, the 2',3'-epoxy ring opens and the 3' position is fluorinated, and at the same time
The hydroxyl protecting groups (cyclohexylidene groups) at the 4″ and 6″ positions are removed, thus
3'-Fluoro-3'-deoxy-6'-N-methoxy (or ethoxy)carbonyl-1,3,3''-tri-N-tosylkanamycin A [referred to as compound (f)]
was found to be generated. The fact that the 3'-position of the kanamycin A compound is fluorinated by the alkali metal hydrogen difluoride along with the opening of the epoxy ring in this way indicates that it has a nucleophilic effect on the epoxy ring of steroids and the epoxy ring of sugars. Fu¨rst−, which predicts the position where a nucleophilic group will be introduced into the ring-opened product when a reaction reagent is applied to open the ring.
Plattner's Law “Advances in Carbohydrate Chemistry and Biochemistry”
Chemistry and Biochemistry), vol. 25, 120-145
Pages, especially pages 123-124 (Published by Academic Press)
(Reference), this is recognized as an abnormal phenomenon. The above compound (f) has its 6'-amino group and 1
When treated with a known deprotection method that removes the amino protecting group at the -, 3- and 3''-amino groups, the desired 3'-fluoro-3'-deoxykanamycin A
(compound) was produced. Also, when the amino protecting group and the hydroxyl protecting group are first removed from compound (e), and then the compound is reacted with an alkali metal hydrogen difluoride under the above-mentioned conditions, 3'-fluoro-
It was confirmed that 3'-deoxykanamycin A () can be produced. In this way, the present inventors succeeded in developing a route for synthesizing 3'-fluoro-3'-deoxykanamycin A starting from kanamycin A. Furthermore, an amino protecting group (tosyl group) on the 1-, 3-, 3″-amino group and an amino protecting group (methoxycarbonyl group, ethoxycarbonyl group, or benzyloxycarbonyl group) on the 6′-amino group of compound (e) group) and the hydroxyl protecting group (cyclohexylidene group) on the 4″- and 6″-hydroxyl groups are removed from compound (e), and the 2′,3′-
Introducing a different type of amino protecting group to each amino group of anhydro-3'-epi-kanamycin A,
The thus obtained 1,3,6′,3″-tetra-N-
When protecting -2',3'-anhydro-3'-epi-kanamycin A is treated with an alkali metal hydrogen difluoride in the same manner as described above, the desirable 1,3,
6′,3″-tetra-N-protected-3′-fluoro-3′-
It was confirmed that deoxykanamycin A can be produced. Starting intermediate compounds used in the synthetic route described above
In (a), the amino protecting group protecting the 1-, 3- and 3″-amino groups is a tosyl group and the 3″- and 6″-
The hydroxyl protecting group that protects the hydroxyl group was a cyclohexylidene group, but these protecting groups can be replaced with other known amino protecting groups and other known hydroxyl protecting groups that work equally well. Even if a protected derivative equivalent to compound (a) is used, the same 4″,6″-di-O-protected derivative as compound (e) can be obtained by performing the same reaction steps as described above.
It has been found that 1,3,3''-tri-N-protected-2',3'-anhydro-3'-epi-6'-N-methoxy (or ethoxy or benzyloxy) carbonyl-kanamycin A can be prepared. The present invention has been completed based on the above-mentioned findings. Therefore, the gist of the first invention is as follows:
The following general formula

[Formula, A is an amino protecting group other than a sulfonyl group type amino protecting group or a hydrogen atom, B is a hydrogen atom or any amino protecting group,
X is a hydrogen atom or a monovalent hydroxyl protecting group, Y is a hydrogen atom or a monovalent hydroxyl protecting group, or X and Y together form one divalent hydroxyl protecting group. shown
2',3'-Anhydro-3'-epi-kanamycin A
or a protected derivative thereof of the following formula MeHF ₂ () [wherein Me is an alkali metal atom] in an organic solvent with an alkali metal hydrogen difluoride of 100 to 200
The following general formula is reacted at a temperature of °C.

[Chemical formula] A 3'-fluoro-3'-deoxykanamycin A compound represented by the formula (wherein A, B, X and Y have the same meanings as above) is produced, and further, a compound of the formula ()
If an amino protecting group and/or a hydroxyl protecting group remain in the compound, these protecting groups are removed by a known method.

A method for producing 3'-fluoro-3'-deoxykanamycin A represented by the following formula: In the first method of the present invention, the general formula ()
2',3'-Anhydro-3'-epi-kanamycin A
The amino protecting group A in the protected derivative may be any known amino protecting group that does not participate in the reaction, such as an acetyl group, an alkanoyl group such as a trifluoroacetyl group, an acyl group such as an aroyl group such as a benzoyl group, a methoxycarbonyl group, or an ethoxy group. an alkoxycarbonyl group such as a carbonyl group or a butoxycarbonyl group; an aralkyloxycarbonyl group such as a benzyloxycarbonyl group or a phenethyloxycarbonyl group; or an aryloxycarbonyl group such as a phenoxycarbonyl group or a methoxyphenoxycarbonyl group; Something can happen. However, if the group A is in the form of an alkyl-, aryl-, or aralkylsulfonyl group such as a mesyl group or a tosyl group, it may participate in the reaction with the alkali metal hydrogen difluoride and produce undesirable by-products. Therefore, the amino protecting group (A) is preferably not of the sulfonyl group type. Although the group (A) can be a hydrogen atom, it is preferable that the 6'-amino group of the compound () is protected. 1-, 3- and in the compound of general formula ()
The 3″-amino group does not need to be protected, but it is an acyl group such as an acetyl group, a benzoyl group, an alkoxycarbonyl group such as a methoxycarbonyl group, an ethoxycarbonyl group, a butoxycarbonyl group;
Aralkyloxycarbonyl groups such as benzyloxycarbonyl group and phenethyloxycarbonyl group; Aryloxycarbonyl group such as phenoxycarbonyl group; Alkylsulfonyl group such as mesyl group, aralkylsulfonyl group such as benzylsulfonyl group; Preferably, it is protected with an amino protecting group such as an arylsulfonyl group. 4″-hydroxyl group in compound () and
Protecting group (X, Y) that protects the 6″-hydroxyl group
can be any known monovalent hydroxyl protecting group, including lower alkyl groups, aryl groups such as phenyl groups; alkanoyl groups such as acetyl groups, or aroyl groups such as benzoyl groups.
However, X and Y may jointly be any known divalent group, including a lower alkylidene group such as an isopropylidene group, an aralkylidene group such as a benzylidene group, a cycloalkylidene group such as a cyclohexylidene group, or a tetrahydropyranylidene group. It may form a hydroxyl protecting group. Introduction of these amino-protecting groups and hydroxyl-protecting groups into kanamycin A compounds has been described, for example, in JP-A No. 55
−105699, British Patent No. 2043634B; Japanese Patent Application Publication No. 1983
−68698, U.S. Patent No. 4357466; Japanese Patent Application Publication No. 1983-
This can be accomplished using conventional protection techniques such as those described in US Pat. No. 152,497 and US Pat. No. 4,359,572. In the first method of the present invention, the alkali metal hydrogen difluoride () to be reacted with the N,O-protected 2',3'-epoxidized kanamycin A compound of general formula () is NaHF ₂ , KHF ₂ or _LiHF2 , but _KHF2 is preferred. This reaction may be carried out in a dry organic solvent suitable for the reaction, but it is preferably carried out in a liquid lower alkylene glycol, preferably ethylene glycol or propylene glycol. Ethylene glycol is most suitable. Depending on the type of organic solvent used, the desired reaction may not proceed. Reaction temperature is 100-200
It can be selected appropriately within the range of ℃, but 120~160
Preferably, it is within the range of °C. The above reaction produces a 3'-fluoro-3'-deoxy-kanamycin A protected derivative of the general formula (), which can be collected by extracting the reaction solution with a suitable organic solvent such as ethyl acetate and then using silica gel.・
This can be carried out by separation and purification using column chromatography. If an amino protecting group and/or a hydroxyl protecting group remain in the compound of general formula (),
The protecting group is removed in the deprotection step. This deprotection step is carried out in stages depending on the type of protecting group used, using a commonly used deprotection method and a known method, such as the method disclosed in the above-mentioned Japanese Patent Application Laid-Open No. It can be done. The reaction solution obtained in this deprotection reaction is concentrated, the solid residue is dissolved in water, passed through a column of CM-Sephadex C-25, and developed with aqueous ammonia to isolate and purify the desired 3'- Fluoro-3'-deoxykanamycin A [compound ()] can be obtained. The gist of the second invention is the following general formula

[In the formula, B' is an amino protecting group, and X and Y
are each a hydrogen atom or a monovalent hydroxyl protecting group, or X and Y are jointly one 2
protection of 3′-O-sulfonyl-6′-N,4′-O-carbonyl-kanamycin A, where Z is an alkylsulfonyl group, an aralkylsulfonyl group, or an arylsulfonyl group] The derivatives are reacted with sodium methylate in the presence of methanol or sodium ethylate in the presence of ethanol or sodium benzylate in the presence of benzyl alcohol to give the following general formula:

[Chemical formula] [In the formula, B', X and Y have the same meanings as above, and A' is an amino protecting group in the form of a methoxycarbonyl group, an ethoxycarbonyl group, or a benzyloxycarbonyl group] 2' , 3'-anhydro-3'-epi-6'-N-alkoxycarbonyl-kanamycin A and then use the compound of formula (') immediately or prepare the amino Protecting group (A′, B′)
and 2',3'-anhydro- obtained after removing part or all of the hydroxyl protecting groups (X, Y)
Using 3'-epi-kanamycin A compound, it is combined with an alkali metal hydrogen difluoride of the following formula MeHF ₂ () [where Me is an alkali metal atom] in an organic solvent at 100 to 200
The following general formula is reacted at a temperature of °C.

[Formula, A″ is an amino-protecting group in the form of a methoxycarbonyl group, an ethoxycarbonyl group, or a benzyloxycarbonyl group, or a hydrogen atom, and B″ is an amino-protecting group or a hydrogen atom,
X' and Y' have the same meaning as X and Y above,
Alternatively, a 3'-fluoro-3'-deoxykanamycin A compound represented by If any protective groups remain, the following formula is characterized in that these protective groups are removed by a known method.

A method for producing 3'-fluoro-3'-deoxykanamycin A represented by the following formula: In the second method of the present invention, 3'-O-sulfonyl- of the general formula () used as a starting compound
The amino protecting group B' and the hydroxyl protecting groups X and Y in the 6'-N,4'-O-carbonyl-kanamycin A protected derivative are each represented by the general formula ( )
They may be the same as the amino protecting group B and the hydroxyl protecting groups X and Y, respectively, in the 2',3'-anhydro-3'-epi-kanamycin A protected derivative. 3'-O-sulfonyl group (Z) has 1 to 1 carbon atoms
The alkylsulfonyl group of 4, such as methanesulfonyl group (mesyl group), ethanesulfonyl group; aralkylsulfonyl group, such as benzylsulfonyl group; and arylsulfonyl group, such as tosyl group, but preferably benzylsulfonyl group. 3′-O-sulfonyl-kanamycin A derivative of general formula () and sodium chloride in the presence of methanol.
Sodium in the presence of methylate or ethanol
The reaction with sodium benzylate (NaOCH ₂ C ₆ H ₅ ) in the presence of ethylate or benzyl alcohol is carried out under anhydrous conditions. Sodium methylate or sodium ethylate or sodium benzylate is a kanamycin A compound ()
It is preferably used in a molar ratio of 2 to 10 per mole of and in a concentration of 1 to 5% by weight in the reaction medium. The reaction temperature can range from 0°C to the reflux temperature of the reaction medium. The reaction time should be at least 5 minutes,
Even if the time is prolonged, undesirable by-products are not produced as much, but the methoxycarbonyl group or ethoxycarbonyl group bonded to the 6'-amino group of the desired 2',3'-epoxidation product ('), etc. A′)
It is preferable to use reaction conditions that do not cause desorption. In the second method of the present invention, the 2,3-epoxidized product of the general formula (') produced as an intermediate is removed, if desired, by removing part or all of its protective groups by a conventional method, The deprotected product may be subjected to reaction with an alkali metal hydrogen difluoride ().
The reaction step of the 2′,3′-epoxidized product of the general formula (′) or its deprotected product with the alkali metal hydrogen difluoride of the formula (), and the subsequent reaction step as necessary. The step of deprotecting the 3'-fluoro-3'-deoxykanamycin A compound of formula (') can be carried out in the same manner as the corresponding step in the first method of the invention. Next, the present invention will be specifically explained with reference to Reference Examples and Examples. Reference example 1 6′-N,4′-O-carbonyl-4″,6″-O-cyclohexylidene-2′,2″-di-O-acetyl-1,3,3″-tri-N- Production of tosylkanamycin A [referred to as compound (b)]

[ka]

[Chemical formula] Compound (a) of the above formula, i.e. 6'-N, 4'-O-
Carbonyl-4″,6″-O-cyclohexylidene-
450 mg of 1,3,3'-tri-N-kanamycin A
(0.427 mmol) in anhydrous dimethyl sulfoxide 2
ml, dissolved in a mixture of 0.23 ml of anhydrous pyridine and N-
188 mg (1.72 mmol) of acetylimidazole was added, and the mixture was stirred at room temperature for 16 hours. 30 ml of 5% sodium hydroxide solution was added to the reaction solution, and the resulting precipitate was collected, washed with water, and then dried. Further washing of the solid with ether and drying yielded 435 mg (90% yield) of the title compound (b).
I got it. Elemental analysis _{Calculated value} ( _{C50H64N4O20S3}・_H2O ) _{: C51.98} ;
H5.76; N4.85; S8.33% Actual value: C51.84; H5.50; N4.72; S8.29% ¹ H-NMR spectrum (pyridine- _d5 ): δ2.22 and 2.23 (respectively) 3H singlet, Ac),
δ2.26, 2.27 and 2.47 (3H singlet, respectively)
NTs(CH ₃ )) Reference example 2 3'-O-pendylsulfonyl-6'-N,4'-O
-Carbonyl-4″,6″-O-cyclohexylidene-2′,2″-di-O-acetyl-1,3,3″-
Production of tri-N-tosyl-kanamycin A [referred to as compound (c')]

[Chemical formula] 77.5 mg (0.068 mmol) of compound (b) obtained in Reference Example 1 was dissolved in 1.5 ml of anhydrous pyridine, and 30.1 mg (0.158 mmol) of benzylsulfonyl chloride was added.
The reaction was carried out at 0-5°C for 10 minutes. After adding 0.1 ml of water to the reaction solution, it was concentrated under reduced pressure. After dissolving the residue in chloroform, the solution was diluted with 5% sodium bicarbonate solution,
It was washed successively with water and dehydrated with anhydrous magnesium sulfate. This solution was concentrated under reduced pressure to obtain 88.0 mg (yield 100%) of the title compound (c'). Elemental analysis Calculated value (C ₅₇ H ₇₀ O ₂₂ N ₄ S ₄・2H ₂ O):
C51.57; H5.62; N4.22; S9.65% Actual value: C51.45; H5.36; N4.33; S9.73% ¹ H-NMR spectrum (pyridine- _d5 ): δ2.23
and 2.25 (respectively 3H singlet, Ac), 2.30,
2.44 and 2.50 (3H singlets, NTs respectively)
(CH ₃ )) Reference example 3 3'-O-benzylsulfonyl-6'-N,4'-O
-Production of carbonyl-4″,6″-O-cyclohexylidene-1,3,3″-tri-N-tosylkanamycin A [referred to as compound (d″)]

[Chemical formula] 668 mg (0.517 mmol) of the compound (e') obtained in Reference Example 2 was dissolved in 10 ml of anhydrous methanol, and 0.4 ml of 2% sodium methylate dissolved in methanol was added.
(0.15 mmol) was added under ice-cooling and reacted for 15 minutes at room temperature. The reaction solution was neutralized with 0.5% hydrogen chloride methanol solution, and then concentrated under reduced pressure. After dissolving the residue in acetone, insoluble matter was removed and the solution was again concentrated under reduced pressure to obtain 546 mg (yield 88%) of the title compound (d″). Elemental analysis Calculated value (C ₅₃ H ₆₆ O ₂₀ N ₄ S ₄ ): C52.72; H5.51;
N4.64; S10.62% Actual value: C52.50; H5.62; N4.58; S10.47% IR spectrum (KBr tablet, cm ^-1 ): 1705 (cyclic carbamate) ¹ H-NMR spectrum (pyridine _-d5 ):
δ2.20, 2.33 and 2.42 (3H singlets each) Example 1 (a) 2',3'-anhydro-4'',6''-O-cyclohexylidene-3'-epi-6'-N-methoxycarbonyl -Production of 1,3,3″-tri-N-tosylkanamycin A [referred to as compound (e′)]

[ka]

[Chemical formula] 600 mg (0.500 mmol) of the compound (d″) obtained in Reference Example 3 was added to 8 ml (2.96 mmol) of 2% sodium methylate dissolved in methanol, and the mixture was heated at 45°C for 30 minutes.
It was allowed to react for a minute. The reaction solution was neutralized with a 0.5% methanol solution of hydrogen chloride, concentrated under reduced pressure, and the residue was dissolved in ethyl acetate, after which insoluble materials were removed. The solution was again concentrated under reduced pressure. The residue was developed with silica gel using chloroform-methanol (50:1 → 10:1).
Separation and purification by column chromatography gave 400 mg (yield 75%) of the title compound (e').
[α] ²⁵ _D +4° (c2.3, chloroform) Elemental analysis Calculated value (C ₄₇ H ₆₂ O ₁₈ N ₄ S ₃・1/2H ₂ O): C52.45; H5.85; N5.20; S8 .93% Actual value: C52.54; H5.78; N5.04; S8.92% (b) 3'-deoxy-3'-fluoro-6'-N-methoxycarbonyl-1,3,3''- Production of tri-N-tosylkanamycin A [referred to as compound (f')]

[C] 174 mg (0.163
mmol) in 3 ml of anhydrous ethylene glycol, and add potassium hydrogen fluoride (KHF ₂ ) to the solution.
Add 90.2 mg (1.16 mmol) and heat at 150-160℃ in a sealed tube.
The reaction was carried out for 3.5 hours (opening of the 2',3'-epoxy ring, 3'-fluorination and elimination of the cyclohexylidene group). Ethyl acetate was added to the reaction solution for extraction, and the extracted solution was washed successively with 5% sodium bicarbonate solution and water, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (developing system: chloroform-methanol, 30:1~
5:1) to obtain 51.4 mg of the title compound (f').
(Yield 31%) and 26.0 mg of by-product (unidentified) were obtained.
[α] ²⁵ _D +3° (c1.3 methanol) Elemental analysis Calculated value (C ₄₁ H ₅₅ N ₄ O ₁₈ S ₃ F・1/2H ₂ O): C48.46; H5.55; N5.51% Actual measurement Value: C48.37; H5.48; N5.48% ¹ H-NMR spectrum (pyridine- _d5 ): δ5.33
(1H double triplet, H-3'; J ₃ ', F ⁵⁵ H ₂ ,
_J2 ' _,3 '= _J3 ' _,4 '=10H2 ₎ (c) 3'-deoxy-3'-fluorokanamycin A
Synthesis of [compound ()]

[Chemical] Dissolve 45.7 ml (0.045 mmol) of compound (f') obtained in the previous section (b) in 3 ml of liquid ammonia at -50 to -60°C, add about 10 mg of metallic sodium, and dissolve at -50 to -60°C.
The reaction was carried out at ℃ for 5 minutes (detosylation). After adding 1 ml of methanol to the reaction solution and concentrating under reduced pressure, the residue was diluted with 5 ml of water.
ml and heated at 50℃ for 15 minutes (in this reaction, 6'-
N-methoxycarbonyl group is eliminated). The reaction solution was cooled to room temperature, 3 ml of Dowex 50W x 2 resin (H ⁺ type) was added, the mixture was adsorbed onto 4 ml of Dowex 50W x 2 resin (NH ₄ ⁺ type), and the target compound was eluted with 0.5N ammonia water. 20.1 mg (yield 91%) of the title compound () was obtained. [α]
²⁵ _D +116° (c0.5 water) This substance has a ^1H -NMR spectrum and an antibacterial spectrum with the 3'-fluoro-3'-deoxykanamycin A substance obtained in the example of the previous patent application No. 161615/1982. Completely agreed on this point. Example 2 2′,3′-anhydro-4″ obtained in Example 1 (a)
，
6″-O-cyclohexylidene-3-epi-6′-N
-methoxycarbonyl-1,3,3″-tri-N-
Tosylkanamycin A, i.e. compound (e')
Dissolve 170mg in 3ml of anhydrous ethylene glycol,
Add 70mg of sodium dihydrogenfluoride (NaHF ₂ ),
The reaction and post-treatment were carried out in the same manner as in Example 1 (b).
47 mg (yield
29%). Example 3 (a) 2',3'-anhydro-4'',6''-O-cyclohexylidene-3'-epi-6'-N-methoxycarbonylkanamycin A [referred to as compound (e ² )]
synthesis of

[ka]

[Chemical formula] 38.5 mg (0.036 mmol) of the compound (e ¹ ) obtained in Example 1 (a), that is, the compound of the above formula, was dissolved in about 15 ml of liquid ammonia at -55 to -60°C, and about 50 mg was added with vigorous stirring. After 15 seconds, ammonium chloride was added to the solution until the blue color disappeared, and the solution was left at room temperature to evaporate the ammonia. 10 ml of water was added and the mixture was concentrated under reduced pressure until the pH reached approximately 9. Amberlite CG-50 for the residue
(NH ₄ ⁺ ) was adsorbed on 5 ml of resin, washed with water, separated and purified with 0.2N ammonia water, and concentrated under reduced pressure.
21.2 mg (yield 97.1%) of the title compound (e ² ) was obtained. ¹ H-NMR spectrum (20% ND ₃ inD ₂ O): δ3.72 (3H, singlet, OCH ₃ ) δ3.85 (1H, double doublet, H-2', J ₁ ' _,2 ' = 3Hz, J ₂ ′ _,3 ′ = 4Hz) 5.10 (1H, doublet, H-1″, J ₁ ″ _,2 ″=4Hz) 5.50 (1H, doublet, H-1′, J ₁ ′ _,2 ′ = 3Hz) ( b) 3'-Fluoro-3'-deoxy-6'-N-methoxycarbonylkanamycin A [Compound (f ² )
generation of

[Chemical formula] 98 mg of the compound (e ² ) obtained in the previous section (a) was dissolved in 3 ml of anhydrous ethylene glycol, 120 mg of potassium hydrogen fluoride was added, and the mixture was reacted in a sealed tube at 150° for 3 hours to dissolve the compound (f ^{2 )} . ) was generated. (c) 3'-fluoro-3'-deoxykanamycin A
Synthesis After removing ethylene glycol from the reaction mixture obtained in the previous section (b) under reduced pressure, the residue containing compound (f ² ) was dissolved in 4 ml of 1N sodium hydroxide, and the mixture was incubated at 80°C for 3 mL.
Heated for an hour (elimination of methoxycarbonyl group).
Transfer the reaction solution to Dowex 50W x 2
(H ⁺ type) was adsorbed onto 20 ml of resin, washed with water, and eluted with 0.5N ammonia water to obtain a substance containing the target compound. This was dissolved in water and placed in a CM-Sephadex C-25 (NH ₄ ⁺ type) column.
The mixture was developed with 0.03-1.5N aqueous ammonia and purified and separated to obtain 21.0 mg (yield 27%) of the target compound. Example 4 (a) 2',3'-anhydro-6'-N-benzyloxycarbonyl-4'',6''-O-cyclohexylidene-3'-epi-1,3,3''-tri-N -Synthesis of tosylkanamycin A [referred to as compound (e ³ )]

[C] 600 mg of the compound (d'') obtained in Reference Example 3 was added to 4
% Sodium Benzilate (

Benzyl Al containing [Formula]) call(

The mixture was added to 10 ml of a mixture of [Formula])-tetrahydrofuran (1:1) and reacted at 45°C for 30 minutes. After neutralization with 0.5% hydrochloric acid and concentration, the residue was dissolved in ethyl acetate, insoluble materials were removed, and the solution was again concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a developing system of chloroform-methanol (50:1→10:1) to obtain 420 mg (74%) of the title compound (e ³ ). (b) Production of 2',3'-anhydro-1,3,6',3''-tetra-N-benzyloxycarbonyl-3'-epi-kanamycin A [referred to as compound (e ⁵ )]

[ka]

[Chemical] 110 mg of the compound (e ³ ) obtained in the previous section (a) is -60
It is dissolved in 40 ml of liquid ammonia at 0.degree. C., and about 150 mg of sodium metal is added with vigorous stirring to cause a reaction, whereby the 6'-N-benzyloxycarbonyl group and tosyl group are eliminated. After 15 seconds, add ammonium chloride until the solution becomes colorless and leave at room temperature.
Ammonia was evaporated. Dissolve the residue in water,
When adsorbed on 3 ml of Dowex 50W x 2 (H ⁺ type) resin (this operation removes the cyclohexylidene group) and eluted with 0.5N aqueous ammonia, the compound of the above formula (e ⁴ ), i.e. A crude product of 2',3'-anhydro-3'-epi-kanamycin A was obtained. This was mixed with water-tetrahydrofuran (1:
2), 25 mg of sodium carbonate and 70 mg of benzyloxycarbonyl chloride (ClCOOCH ₂ φ) were added, and the mixture was reacted at room temperature for 3 hours to introduce a benzyloxycarbonyl group (Z) into the amino group. The reaction solution was concentrated under reduced pressure, water was added, and the precipitate was collected to obtain 84 mg (87%) of the title compound (e ⁵ ). (c) 3'-fluoro-3'-deoxykanamycin A
Synthesis of [compound ()]

[Chemical] 90 mg of the compound (e ⁵ ) obtained in the previous section (b) was dissolved in 3 ml of anhydrous ethylene glycol, 50 mg of potassium hydrogen fluoride (KHF ₂ ) was added, and the mixture was allowed to react in a sealed tube at 150° for 3 hours. Ta. Thereafter, treatment was carried out in the same manner as in Example 1 (b) to obtain the compound (f ³ ) of the above formula, that is, 1,3,6',3''-tetra-N-benzyloxycarbonyl-3'-fluoro- A crude product (24 mg) of 3'-deoxykanamycin A was obtained. It was diluted with water-dioxane (1:1) without purification.
ml, palladium black was added, and reduction was performed under 3 atmospheres of hydrogen to remove the benzyloxycarbonyl group (Z). Dissolve the resulting substance in water,
CM-Sephadex C-25 (NH ₄
⁺ type) was developed with 0.03-1.5N ammonia water and purified to obtain 9.5 mg (yield 22%) of 3'-fluoro-3'-deoxykanamycin A. Example 5 (a) 2',3'-anhydro-3'-epi-1,3,6'
，
Synthesis of 3″-tetra-N-acetylkanamycin A [referred to as compound (e ⁶ )]

[Chemical formula] Compound (e ³ ) obtained in Example 4 (a), i.e.
2',3'-Anhydro-6'-N-benzyloxycarbonyl-4'',6''-O-cyclohexylidene-
Using 110 mg of 3'-epi-1,3,3''-tri-N-tosylkanamycin A, it was treated in the same manner as the deprotection treatment in Example 4 (b) to give 2',3'-anhydro- 3′-epi-kanamycin A [compound (e ⁴ )]
A crude product was obtained. This crude compound (e ⁴ ) was suspended in 2 ml of water-tetrahydrofuran (1:2) and 45 mg of acetic anhydride was added in the presence of 25 mg of sodium carbonate.
was added and reacted at room temperature to protect the amino group of compound (e ⁴ ) with an acetyl group. The reaction solution was concentrated under reduced pressure, and the resulting solid was water-methanol (1:
1), 1 ml of Dowex 50W (H ⁺ type) resin was added to the solution, stirred, and filtered. Combine the solid washing liquid and liquid,
The mixture was concentrated to dryness under reduced pressure to obtain 80 mg of the crude title compound (e ⁶ ). (b) 1,3,6′,3″-tetra-N-acetyl-
3'-Fluoro-3'-deoxy-kanamycin A
[Production of compound (f ⁴ )] The crude compound (e ⁶ ) obtained in the previous section (a) was dissolved in 3 ml of anhydrous ethylene glycol, 50 mg of potassium hydrogen fluoride (KHF ₂ ) was added, and the mixture was placed in a sealed tube. The reaction was carried out at 150° for 3 hours. Thereafter, treatment was carried out in the same manner as in Example 1 (b) to obtain the title compound (f ⁴ ), i.e. 1,3,6′,
3″-tetra-N-acetyl-3′-fluoro-3′-
20 mg of crude deoxykanamycin A was obtained. (c) 3'-fluoro-3'-deoxykanamycin A
Synthesis of the crude compound (f ⁴ ) obtained in the previous section (b)
Dissolve in 1 ml of 2N-sodium hydroxide aqueous solution,
C. for 2 hours to deacetylate. After that, it was adsorbed to 1.5 ml of Dowex 50W x 2 resin (H ⁺ type) and eluted with 0.5N ammonia water to obtain the target.
9.7 for 3'-fluoro-3'-deoxykanamycin A
I got mg. In the formulas shown in the reference examples and examples above, Ts represents a tosyl group, Ac represents an acetyl group, Bzs represents a benzylsulfonyl group, Cbm represents a methoxycarbonyl group, and Z represents a benzyloxycarbonyl group.

Claims (1)

[Scope of Claims] 1 The following general formula: [In the formula, A is an amino-protecting group other than a sulfonyl group-type amino-protecting group, or a hydrogen atom, and B is a hydrogen atom or any amino-protecting group. is the basis,
X and Y are each a hydrogen atom or a monovalent hydroxyl protecting group, or X and Y are jointly 1
2',3'-Anhydro-3'-epi-kanamycin A or its protected derivative represented by the following formula MeHF ₂ () [wherein Me is an alkali metal 100 to 200 in an organic solvent with an alkali metal hydrogen difluoride of
℃ to produce a 3'-fluoro-3'-deoxykanamycin A compound represented by the following general formula: [wherein A, B, X and Y have the same meanings as above] and then the expression ()
If an amino protecting group and/or a hydroxyl protecting group remain in the compound, the 3'-fluoro- Method for producing 3'-deoxykanamycin A. 2. Claim 1, wherein the organic solvent used in the reaction of the 2',3'-anhydro-3'-epi-kanamycin A compound () and the alkali metal hydrogen difluoride is a liquid lower alkylene glycol. the method of. 3. The method according to claim 1, wherein the alkali metal hydrogen difluoride is sodium hydrogen difluoride or potassium hydrogen difluoride. 4 The following general formula: [In the formula, B' is an amino protecting group, and X and Y
are each a hydrogen atom or a monovalent hydroxyl protecting group, or X and Y are jointly one 2
protection of 3′-O-sulfonyl-6′-N,4′-O-carbonyl-kanamycin A, where Z is an alkylsulfonyl group, an aralkylsulfonyl group, or an arylsulfonyl group] The derivative is reacted with an alkali metal methylate in the presence of methanol or an alkali metal ethylate in the presence of ethanol or sodium benzylate in the presence of benzyl alcohol to form the following general formula: has the same meaning as above, and A' is an amino protecting group in the form of a methoxycarbonyl group, an ethoxycarbonyl group, or a benzyloxycarbonyl group. A protected derivative of N-alkoxycarbonyl-kanamycin A is generated and then a compound of formula (') is used immediately or a compound of formula (') is prepared with amino protecting groups (A', B') and hydroxyl protecting groups ( 2′,3′-anhydro- obtained after eliminating part or all of X, Y)
Using 3'-epi-kanamycin A compound, it is combined with an alkali metal hydrogen difluoride of the following formula MeHF ₂ () [where Me is an alkali metal atom] in an organic solvent at 100 to 200
The reaction is carried out at a temperature of 3'-fluoro-3, which is an amino protecting group or a hydrogen atom, and X' and Y' have the same meanings as X and Y above, or X' and Y' are each a hydrogen atom] '-Deoxykanamycin A compound is produced, and if an amino protecting group and/or a hydroxyl protecting group remain in the compound of formula ('), these protecting groups are removed by a known method. A method for producing 3'-fluoro-3'-deoxykanamycin A represented by the following formula: 5. According to claim 4, the organic solvent used in the reaction of the 2',3'-anhydro-3'-epi-kanamycin A compound (') and the alkali metal hydrogen difluoride is a liquid lower alkylene glycol. Method described. 6. The method according to claim 4, wherein the alkali metal hydrogen difluoride is sodium hydrogen difluoride or potassium hydrogen difluoride.