JPS61143393A - 2',3'-dideoxy-2'-fluorokanamycin a - Google Patents

Abstract

NEW MATERIAL:2',3'-dideoxy-2'-fluorokanamycin A shown by the formula I and its acid addition salt. USE:An antibacterial agent. showing high antibacterial activity also against bacteria resistant to kanamycin. PREPARATION:For example, a compound [e.g., 6-0-(2-0-acetl-4,6-0-cyclohwxyli dene-3-deoxy-3-tosylamino-alpha-D-glucopyranosyl)-1,3-bis-N-tosyl-2-de -oxystreptamine, etc.] shown by the formula II (Y is NH2-protecting group; B and D are OH- protecting group) is reacted with a compound(e.g., 6-azido-4-0-benzoyl-2,3- dideoxy-2-fluoro-ribo-hexopyranosyl bromide, etc.) shown by the formula III (A is OH-protecting group) in an anhydrous organic solvent (preferably dichromomethane) at 80-100 deg.C, the azido group is reduced, and deprotected.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides a novel semi-synthetic aminoglycoside antibiotic, λ'
, 3'-dideoxy-λ'-fluorokanamycin A. This new compound exhibits high antibacterial activity against various kanamycin-sensitive and kanamycin-resistant bacteria and is useful as an antibacterial agent.

Kanamycin A as a semi-synthetic aminoglycoside antibiotic derived from kanamycin A, B and 0.

Various deoxy derivatives of B and O are known.

These conventionally known deoxykanamycin derivatives have useful antibacterial activity, but their antibacterial spectrum varies widely, and new antibacterial agents may emerge that may render them ineffective. There is a constant desire to create chemical compounds. The present inventors believe that if they can create a derivative in which the hydroxyl group at the J' position of kanamycin A is replaced with a fluoro group, that is, J'-fluoro-3'-deoxykanamycin A, this will be a compound that is effective against kanamycin-resistant bacteria. We expected that this would be the case, and as a result of our research, we succeeded in synthesizing it. Namely, the present inventors have proposed l-fluoro-3 as a novel semi-synthetic minoglycoside antibiotic in the earlier Japanese Patent Application No. Shoj F-141411 (filing date: January 2008-1999). '-Deoxykanamycin At-
J-Fluoro-/,ni! , t-di-0-isozolobylidene-α-D-Dalkov 2-north, and a multi-step route to synthesis method.

Furthermore, this time, the inventor has determined that %21,41-dideoxy-
Research was conducted with the intention of creating 2'-fluorokanamycin At, and as a result of the research, [Oan, J.

Ohem, J 4'vol. 726-7P.
), the known substance methyl a, t-o-pendycudene-J-deoxy-β-D-arabino-hequinpyranoside and the known substance &-0-(J-amino-J-deoxy-α-D-glucopyranosyl )-2-deoxystreptamine, Eri-2', J'-)thioxy-λ'-fluorokanamycin A'v1- was synthesized for the first time using the following synthetic route as shown in the Reference Examples and Examples described below. Moreover, this new compound was successfully used in various Durham-positive bacteria, including mt-resistant.

It was confirmed that it has antibacterial activity against negative bacteria.

Therefore, the key feature of the present invention lies in λ', 3'-dideoxy-2'-fluorokanamycin A and its acid addition salt represented by the linear formula.

The λ', J'-dideoxal λ'-fluorokanamycin of the present invention is a white powder with no definite melting point and is a basic substance exhibiting the physical properties described in the Examples below. The novel compounds of formula (i) of the present invention are typically.

Obtained as free base or hydrate or carbonate, but 1
Any acetic acid addition salt can be prepared in a conventional manner. Examples of acid addition salts include pharmaceutically acceptable inorganic acids such as hydrochloric acid, sulfuric acid, 11-acid, nitric acid, or acetic acid;
There are salts with pharmaceutically acceptable organic acids such as malic acid, citric acid, ascorbic acid, and methanesulfonic acid.

The invention! , J'-,) Deoxy-λ'-fluorokanamycin A or its acid addition salt can be formulated into an antimicrobial composition by combining it with a pharmaceutically acceptable liquid or solid carrier.゛The antibacterial spectrum (minimum inhibitory concentration) of λ', J'-dideoxy-λ'-fluora kanamycin A (abbreviated as 21,31-dideoxy-λI-v-kanamycin A) (free base) of the present invention is as follows: As shown in Table 1. For comparison, the antibacterial spectrum of Kanamycin A is also shown in Table 1.

As mentioned above, one of the present invention's λ', J'-dideoxy-λ'
-Fluorokanamycin A has higher antibacterial activity than kanamycin A against resistant bacteria. This is true for both resistant E. coli and secondary Pseudomonas aeruginosa.

The method for producing λ', J'-dideoxy-λ'-fluorokanamycin A of the formula (Il) of the present invention, apart from the raw material preparation step, involves the step J of the arrow, that is, the following formula [where A is a hydroxyl protecting group]. 1 For example, a t-dideoxy group or an aralkyl group, preferably a benzoyl group or a benzyl group.
-Fluororibohexopyranosyl bone bromide t-
[In the formula, Y is a hydroxyl-protecting group such as a hydroxyl-protecting group, such as a ryokyloxycarbonyl group, an alkyloxycarbonyl group, an alkylsulfonyl group, an arylsulfonyl group, or an aralkylsulfonyl group, and %D is a hydroxyl-protecting group, such as an acyl group. and B is a monovalent hydroxyl protecting group, each different or the same as D, such as an acyl group or an aralkyl group, or λ B's jointly serve as a divalent hydroxyl protecting group, an alkylidene group or a cyclo of an alkylidene group or an aralkylidene group /fakR]
J'-N-protected-J'-amino-J'-deoxy-a-
〇-glucopyranosyl)-/, j-bis-N-protected-2
-condensation with the φ-position hydroxyl group of deoxystreptamine to form the following formula [wherein A, B, D and Y are the same protecting groups as above]
A step of producing a condensation product represented by

A step of reducing the cytogroup (N5) of the condensation product of formula (IV) and converting it into an amine group, and converting the amination product to the remaining protecting groups (A, B, D, Y): and a step of eliminating the target compound of formula (1) by a conventional method, namely, λ',3'-dideoxy-λ'-fluorokanamycin A'l-.

The formula of the raw material (the starting compound of 10 is % Reference Example 1 (a) described below)
6-Azide-〇-benzoyl produced by the method shown in 2. λ-fluoro-D-ribo-hexopyranose in J, 4-trideo is prophylated by reacting its 1-position hydroxyl group with thionyl bromide to give t-azido-4cm0-benzoyl λ, J, 4 -trideogysicofluoro-2-ribo-hexopyranosyl bromide (see Reference Example 1 (e)) 1) Then, if desired, the Hiro-0-benzoyl group is converted to other hydroxyl protecting groups (A1 e.g. benzyl group) by known methods. It can be prepared by replacing it with L.

The compound of formula (In) is a known substance t-〇-(Jf
-amino-3'-deoxy-α-D-glucopyranosyl)-codeoxystreptamine at position 1.

3-position The 3-position amino group is protected with a known 7-mino group (Y), such as a tosyl group, and then the 22-position hydroxyl group is protected with a known hydroxyl group (D), such as an acetyl group. -(λ'-〇-Protection-31-N-Protection-
Using cystrezotamine in the 3'-amino-J'-deoxy-α-D-glucopyranosyl)-t, J-bis-N-protected M-2-deo position.

Hydroxyl group at position 1 - known monovalent hydro Φ
mm can be obtained by protecting with a syl-protecting group (B) or a known covalent hydroxyl-protecting group such as an alkylidene group 1 such as an inzolobylidene group, an aralkylidene group 1 such as a benzylidene group, or a cycloalkylidene group 1 such as a cyclohexylidene group.

The order of introduction of protecting groups can be adjusted as convenient. The compound of formula (n) is t-〇-(
-10-acetyl ginseng, t-0-cyclohexylidene-
J-y"oxy-J-)silucodeoxystreptamine α-D-glucopyranosyl)-i, J-bis-N-1silucodeoxystreptamine, or the reference example in the specification of the patent application No. A-0-(-10-benzyl-J-penzyloxycarponylamino-44, A
-0-cyclohexylidene-J-deo-α-D-/
(rucobylanosyl)-t, J-bis-N-penzylocyclocarbonyl-co-deoxystreptamine.

The condensation step between the compound of formula (If) and the compound of formula (Ill) is:

It may be carried out in the presence of mercuric cyanide as a catalyst in an anhydrous co-solvent 1, for example a chlorinated hydrocarbon, preferably dichloromethane, under heating at a temperature of, for example, 10 DEG to 100 DEG C. The azide group (N5) of the resulting condensation product of formula (fV) can be reduced by a known method, for example, by catalytic reduction in the presence of a palladium catalyst. Reduction of the azide group is performed simultaneously with removal of the protective group. For example, when the amine protecting group (Y) is a tosyl group, when this tosyl group is deprotected by treating with liquid sodium monoxide metal sodium,
The azide group is also reduced to an amine group by the reduction action that accompanies this process (see Example 1 (b)). The remaining protective group (A.

The deprotection step for eliminating B, Y) can be carried out in one step or in stages according to the type of protecting group used, using a commonly used deprotection method and a known method. In addition.

The above condensation reaction and reduction reaction of the azide group are 3'-deoΦ
U.S. Patent No. J, P 2 for Synthesis of Cikanamycin Man
F, 7 A It can be carried out in the same manner as shown in the specification of No. 7.

The reaction solution obtained in this deprotection reaction was concentrated, the solid residue was dissolved in water, and CM-Sephadex C-λ! The target z,i'-dideoxy-2'- of formula (1) is isolated and purified by passing it through a column of
Fluorokanamycin A can be obtained.

Next, reference examples and examples of the present invention will be specifically explained. In the formulas shown in these Reference Examples and Examples, %MS represents a methyl group, Ph represents a phenyl group, Bz represents a benzoyl group, Ts represents a tosyl group, and Ac represents an acetyl group.

Reference Example 1 (a) Methyl@μ,6-0-benzylidene-J-deoxy-ID-arabino-to bovine sopiranoside [Compound (1
) from methyl hiro, z-0-benzylidene-2
, J-dideoxy-2-fluoro-β-D-ribo-hexopyranoside [Compound (referred to as 24)] To a mixture of anhydrous benzene, Jjd, and anhydrous pyridine iH was added 1.1 tIl of diethylaminosulfatrituruolide, and the mixture was heated at 0°C. Under stirring, add known substances (E, H).

Williams, W.A., 8zarek, a.
nd JaK, N. Jones: 'Oan, J. Ohe.
m, ', 4'ri, 726-72F (/ri7/))
200 μ of compound (1) was added and dissolved,
After stirring at 0°C for 0.2 hours, the mixture was heated at 10°C for 3 hours. The reaction solution was cooled on ice and fc11i! l hydrogen carbonate)
IJum aqueous solution - 2!0 tdlfC was added and stirred vigorously for 03 hours. Thereafter, benzene too and z2 were added, extracted, washed with water, dried over anhydrous sodium sulfate, and concentrated to obtain a syrup. The product was purified by silica gel column (/ρ0Ml!, Lycium; developed with holph) to obtain compound (2) as crystals. Yield 7
1%6 (b) Compound (2) Nidimethyl-4C-0-hencyyl-t-bromo-2,! ,,4-trideoxy-cofluoro/-D-ribo-hexopyranosite [Compound (3
) Synthetic compound (3) of compound (2) in the previous section (a), N-bromosuccinimide λtrap, Nororium carbonate 447
To 4C Ki.

Anhydrous carbon tetrachloride λO- was added and the mixture was heated under reflux for 1 hour. The reaction suspension was concentrated, extracted by adding aaa foam, and after filtering off insoluble matter, the extract was washed with water.

It was dried over 13 um of anhydrous sulfuric acid and concentrated to give the crude compound (3) Datata Misaki as pale yellow crystals. Roughness rate: 22%.

(c) Synthesis of methyl 6-azido-hiro-〇-penzoyluko, 3,6-drideoxy-cofluoro/-D-ribo-hexopyranosite [referred to as compound (4)] from compound (3) Compound (4) (referred to as compound (4)) Crude product 4CFPsy of compound (3) obtained in (b)
(Dissolved in 10wt of anhydrous dimethylformamide, added sodium azide/2saki, and reacted at %toyC for O0j time. After concentrating the homogeneous reaction solution, 10wt of chloroform
The extract was washed with water and dried over anhydrous sodium sulfate to obtain a yellowish brown syrup. This was purified by silica gel column chromatography (JOgd, chloroform) to obtain a colorless syrup compound (4 ciOqt).

The yield of compound (2) was 6 to 0)' From compound (4), t-diy-u-o-benzoyl-4,J,≦-tridiofluoro-D-ribo-hexopyranose [ A mixture of trifluoroacetic acid and wax (/ : / ) containing 1lttqf & normal hydrochloric acid of the compound (4) of the preceding item (c),
6- and reacted at ro°C for 1 hour. The reaction solution was diluted with water, neutralized by adding sodium bicarbonate under water cooling, and then extracted with chloroform.

The extract was washed with water, dried over anhydrous sodium sulfate, concentrated to obtain a syrup, and purified by silica gel column chromatography (JM, toluene = ethyl acetate = A:l) to obtain a colorless syrup of metal compounds ( 1IOq of 5) was obtained. Yield 20%.

(e) Synthesis of sopyranosyl bromide [referred to as compound (6)] from compound (5) to t-azido-44-0-pencyyl-, J-dideomalcy-fluoroligoe (referred to as compound (6)) Compound (5) Nore 4471F'i was dissolved in anhydrous dichloromethane-25'd to convert Pdi to thionyl bromide.
The mixture was added and reacted for 34 hours at room temperature. Add hydrogen carbonate to the acidic reaction solution) Add IJ trott (after neutralization).

Extract with dichloromethane, filter off insoluble materials, and concentrate.
Syrup t-obtained. It was purified by Korekura silica gel column chromatography (JjOmb benzene) to obtain 1.0 Jtt of compound (6) as a colorless syrup. yield! t%
.

Optical rotation: [α)''-+/Ib° (c' e chloroform) Infrared absorption spectrum; Koi i Oon-'(7d)'H-NM 几spectrum (in deuterated chloroform, tetramethylsilane internal standard) : δ2.J2 (/l(, dqH-jax), -2j4A
(/H.

dddt H-J eq) u, A j (/H,
dddd H-λ).

! , l4c(/H, ddddf(-41,j, 4/ (
/H.

d H-/) '1.2= ""J2,5ax='λ,0,
J2.5e, = 44.7.

2J5ax, 5eq = '''' * J5ax,
4 = "λ, 0゜5J5aq, 4 = '-' *
Jl, 4-~/ , J/, F -6-~Q 2J2 , F = 4' 7・!・ '5ax, F
= ”” J5eq, F' = yo.

'J4. v Mi/-j Hz Reference example 2 (a) 4-O-(j-amino-J-deomushi α-
D-glucopyranosyl)-codeoxystreptamine [referred to as compound (7)]L4-O-(J-deoxy-
j-) cylamino-α-D-glucopyranosyl)-t,
Synthetic compound (7) of i-bis-N-tosylu-deoxystreptamine [referred to as compound (8)]
Compound (8) Known compound (7) /! , 0? and sodium carbonate, io, or dioxane-water (/:/
) was dissolved in 5OOd, 27.7v of tosyl chloride was added under water cooling, and after stirring for 1 hour, it was left at room temperature overnight (/4L).
time) reacted (N-tosylation). The residue obtained by concentrating the reaction solution was extracted with ethyl acetate, and after washing the extract with water,
It was dried over anhydrous sodium sulfate and condensed to obtain colorless solid compounds (8) JP and JP'ii. Yield 1r1% using compound (7) as free pace.

(b) From compound (8), 4-0-(but-0-cyclohexylidene-3-deo)
-D-glucopyranosyl)-t,i-bis-N-)sil-codeoxystrezotazine [referred to as compound (9)]
Synthetic compound (9) of compound (8) of the previous section (a) -! ,0ft-Anhydrous N,N
-Dimethylform completed) 11JOOgI! Dissolve NL, Nog 2 in toluenesulfonic acid 1. lf and/,/-dimethoxycyclohexane j, Jtdk processing, ≦O'C%20~λ
jwaH? 1 hour under the pressure of
41-O-cyclohexylidene).

Reaction solution t-j% carbonated water, sodium aqueous solution λ! In addition to
The resulting precipitate was collected and purified by silica gel column chromatography (developed with chloroform-methanol-t:i) to give a colorless solid compound (9).
I got f. yield! At the same time, 1.7 f (/FtX) of di-0-dichlorohexylidene compound was also obtained.

(c) Compound (9), 4-0-(co-0-acetyl-4.j-0-cyclo, cylidene-J-deoxy-J
-tosylamino-α-D-/rucobyranosyl)-t, J
Synthesis of -bis-N-)silylcodeoxystreptamine [referred to as compound αO] 0^1- Compound αG Compound 'to) of the previous section c) l≠, u? f Anhydrous dimethyl sulfoxide-anhydrous pyridine (9:t) mixture 7
0d, N-acetylimidazole J and 74r gold were added, and after replacing with nitrogen, the mixture was reacted at room temperature for 27 hours (λ
′-〇-acetylation).

The reaction solution was saturated with hydrogen carbonate) IJ cum aqueous solution -! In addition, the resulting precipitate was washed with water, further washed with ethyl ether, and subjected to silica gel column chromatography (λA,
Compound (10) was purified as a colorless solid (developed with chloroform-methanol/IO:l) to give compound (10)/0. / r'i
Obtained. Yield: 47Y.

Optical rotation = [α) ” + 7'(c' *chloroform).

[α)” +41!” (e/ ,DMF)'H-NM
R spectrum (in deuterated pyridine, tetramethylsilane internal standard): δ Co, Co J, Co, 26. Ko, Ko I (J Hs, o
h5. o6H4゜so□-), 2.4' 4 (j H
s, OH5,00-), j, At(/H,d
d, H-,, z'), t,, z Hiro(/Hd, H-
/') 3J - JJ Hz 6J2t 5t =
/ 0.1 Hzl, 2" (a) Co1-〇-7ceteru-t'-azido-zhen'-〇-pensoyl-4ct, y + o-cyclohexylitine-t'-deamino-λI, Jl -, ) Synthesis of 2'-FluorotholyN-tosylkanamycin A [Compound aυ] Compound (6) obtained in Reference Example 1 was dissolved in 1.J- of anhydrous dichloromethane and pulverized. ire to dry light, crushed mercuric cyanide J-4/-jI1
9t-, and here the compound aatJ obtained in Reference Example
Add R Misaki and dissolve at 1 aoC, protected from light.

The mixture was stirred for 7 hours (condensation reaction). The reaction suspension 1- was diluted with chloroform (1OOIId), filtered using Celite, and the filtrate was washed with a saturated aqueous sodium bicarbonate solution and water in turn, dried with anhydrous sulfuric acid, and dried with IJum.
Shrink. The obtained brown solid gold was purified by silica gel column chromatography (/JOd, benzene; ethyl acetate = to
: When separated and purified using 9), pale yellow solid compound (6) was obtained.
The crude JOO cape and the %μ-condensed compound Q3-λj1q
(yield 27% from compound (to)) was obtained.

Crude compound a1) ioOqt - silica gel column chromatography again (40 m, benzene:ethyl acetate=S
: I of the desired compound (b) purified in 3) as a colorless solid
Obtained JIAq. Yield/A% of compound αOnid.

(b) Compound (b) Technique #) λ', J'-dideoxy-2
'-Fluorokanamycin [among compounds] Synthetic compound II) Compound θυ/J7q of the previous section (a) was treated with Sephadex L.
H-20 KAMKU AR Togerahui (/7td.

Colorless solid compound Q obtained by repurification with acetone)
l)/J 4(, ko"if, -jOCICkvs, add 200q of dissolved metal sodium to the liquid and react by stirring at -10°C for 4 hours. 9. Reduction of azide group and When the tosyl group was eliminated, about 11 df of methanol was added to turn the dark-colored reaction solution into pale yellow.
Concentration gave a pale yellow residue aI. When this is dissolved in water, the lyacetyl group and benzoyl group can be eliminated by flukaline hydrolysis. 7nonono-')ite OG-/20 resin (H+ type λoo~4co.

mesh) λ,,r Of t-addition, neutralization, resin t-
Take F.

The reaction solution was washed with water, developed and eluted with 1/7 mL of water, and the eluate with positive coloration was collected and concentrated to obtain a pale yellow solid. When this solid was dissolved in 10 d of a mixture of acetic acid and water (4C:/) and heated at 10° C. for 0.2 hours, the cyclohexylidene group was eliminated. The solid obtained by concentrating the reaction solution was dissolved in water, passed through an OM-Sephrodex 0--I column (J (7d), and the ammonia att- Developed while gradually increasing.The eluate was colored with ninhydrin, and the part containing the target substance (77~rijm)f: was collected and concentrated as a colorless solid.T'--yy'oxy-! - 1jlA,/q of fluorokanamycin A was obtained (23% yield as H2O05 salt).

Optical rotation! f: [α) + ” 6 @(c (' J
e water)'H-NMR spectrum (20% deuterated ammonia-
in heavy water, tetramethylsilane internal standard): d-, 7/(/H, ddtH-1'), z, z≠ (/
H, d.

H-i') 5J1/, 21 J, j, Jll, v-
~0. J2/ , v = Hiroshi 1. OHz Reference Example J Deprotection of β-condensation compound (2) obtained in Example 1 (a).

Compound (2) Compound 1 Example 1 (a)
The co-produced compound 0, RG and Sephadex L
Colorless solid compound Q3λ obtained by repurification by H-λθ column chromatography (co-Od, acetone development) K
00. twpt-, at -10°C, metal sodium JOOqf dissolved in 4 cOwd of solution ammonia was added, and the mixture was stirred at -J-0'Q for 2 hours. About methanol! - was added, the dark blue reaction solution turned pale yellow, and concentrated.
A pale yellow residue was obtained. Dissolve this in water 20114 and add An''-La()CG-/20 resin (H+Il1% aoo ~
Go.

Add mesh) -027ft- to neutralize and remove resin'kF.

Wash with water, develop and elute with 1N aqueous ammonia.

The ninhydrin-positive eluate was collected and concentrated to obtain a pale yellow solid. This solid was mixed with acetic acid-water (4c:/),
1 and heated at roC for 0.3 hours and then concentrated. The obtained solid was dissolved in water and OM-Sephadex O-co! column (jOd), 0 to a, /! The ammonia was gradually increased in aqueous ammonia from normal ammonia water. The eluate was colored with ninhydrin, and the target substance α4t-
The containing portion (4'0'-6Od) was collected and concentrated to obtain 9 compounds 04)'OJ4'sv represented by the above formula as a colorless solid. Yield core pigeon procedure amendment (voluntary) April 25, 1985

Claims (1)

[Claims] 1. 2',3'-dideoxy-2'-fluorokanamycin A and its acid addition salt represented by the following formula ▲ Numerical formula, chemical formula, table, etc. ▼ (I).