CH648564A5 - IF NECESSARY, 1-N-ACYLATED 5,3,4-TRIDEOXY- OR 5,3,4-TRIDEOXY-6-N-METHYL- OR 5,3,4,6-TETRADEOXYKANAMYCIN-B AND METHOD FOR THE PRODUCTION THEREOF. - Google Patents

Description

The present invention relates to new semi-synthetic aminoglycosidic antibiotics which contain an IN - [a-hydroxy-co-aminoalkanoyl] -5,3 ', 4'-trideoxykanamycin-B, 1 -N- [a-hydroxy-co-aminoalkanoyl] -5,3 ', 4', 6 "-tetradeoxyka-namycin-B and 5,3 ', 4', 6" -tetradeoxykanamycin-B and an IN- [a-hydroxy- (o-aminoalkanoyl] -5,3 ', 4'-trideoxy-6'-N-methylkanamycin-B and 5,3', 4'-trideoxy-6'-N-methyl-kanamycin-B, all of which are new compounds useful as antibacterial agents "The invention further relates to methods for producing these new compounds. Finally, the invention further relates to an antibacterial preparation which contains one of these new compounds as an active ingredient. ***"

Dibekacin, that is, 3 ', 4'-dideoxykanamycin-B was produced semi-synthetically from kanamycin-B, as described in Japanese Patent Laid-Open No. 7595/75, Japanese Patent No. 794 612 and U.S. Patent No. 3,753,973. Dibekacin has been used extensively for the therapeutic treatment of various bacterial infections as a chemotherapeutic agent which is effective against the kanamycin sensitive bacteria and also against various kanamycin resistant bacteria. Habekacin was also produced semi-synthetically, namely 1-N- (L-4-amino-2-hydroxybutyryl) -dibe-

kacin, which is a chemotherapeutic agent effective against dibekacin resistant bacteria (see Japanese Patent Laid-Open No. 33629/77, U.S. Patent No. 4,107,424). Furthermore, an IN- [a-20 hydroxy-co-aminoalkanoyl] -6'-N-methyldibekacin was also prepared semisynthetically, which was found to be highly effective against various strains of bacteria (Japanese Patent Application No. 115 199/74, British Patent No. 1,475,481, U.S. Patent No. 4,147,861). In further research by the same inventors, the 6 "-deoxy or 4", 6 "-dideoxy derivatives of dibekacin and habekacin were synthesized, ie of 3 ', 4'-dideoxykanamycin-B and of lN- (L-4- Amino-2-hydroxybutyryl) -dibekacin Furthermore, it was found that these 6 "-deoxy derivatives and 4", 6 "-dideoxy-30 derivatives of dibekacin or habekacin not only have a low ototoxicity but also an antibacterial activity which is just as high as Dibekacin or Habekacin (see Japanese Patent Laid-Open No. 119 323/79, British Patent Laid-Open No. GB 2 058 774 A and US-35 Patent No. 4332 794).

As a further deoxy derivative of dibekacin, 5,3 ', 4'-trideoxykanamycin-B was produced (Japanese Journal of Anti-biotics, 32, p. 178 [1979]). However, 5,3 ', 4' -trideoxykanamycin-B was found to be less effective than Dibe-40 kacin.

Furthermore, some deoxy derivatives of IN (L-4-amino-2-hydroxybutyryl) kanamycin-A (ie, amikacin) were prepared including 3'-deoxyamikacin (Japanese Patent Laid-Open No. 49 105/75, U.S. Patent No. 45 4 104372), 3 ', 4'-dideoxyamicacin (Japanese Patent Laid-Open No. 11 402/79, British Laid-Open No. GB 2 043 034 A, U.S. Patent No. 4,298,727). 6 "deoxy-amikacin (Japanese Patent Laid-Open No. 54 733/79, 4", 6 "-Dideoxyamikacin (Japanese Patent Laid-Open No. 54733/79), 3 ', 4', 4", 6 "-tetradeoxyamicacin (Japanese Patent Laid-Open No. 138 685/79), and 3 ', 4', 6 "-tride-oxyamikacin (Japanese Patent Application No. 5 657/80, as well as 3 ', 6" -dideoxyamikacin, 5,3'-dideoxyamikacin and 5,3' , 6 "-trideoxyamikacin (Japanese laid-open publication ss No. 107 202/80, all of which have a low acute toxicity and a low ototoxicity as well as a useful antibacterial activity as high as that of amikacin.

Continuing to pursue research on 60 deoxy derivatives of dibekacin and habekacin, a new compound, 5,3 ', 4', 6 "-tetradeoxykanamycin-B, was found which was found to be remarkably effective against certain types of bacteria, despite the fact that the 5th , 3 ', 4'-trideoxykanamycin-B does not have a useful antibacterial activity. Furthermore, a new compound was successfully produced, namely 5,3', 4'-trideoxy-6'-N-methylka-namycin-B, which also has a higher antibacterial activity than that of 5,3'-4'-trideoxyca

648564

10th

namycin-B, especially against certain resistant strains. In an effort to develop such new derivatives of 5,3 ', 4'-trideoxykanamycin-B, 5,3', 4 ', 6 "-tetradeoxykanamycin-B or 5,3', 4'-trideoxy-6'-N-methylkanamycin -B, which have an improved antibacterial activity, have now been developed new IN- [a-hydroxy-co-aminoalka-noyl] derivatives of these deoxykanamycin-B compounds by acylation of the 1-amino group of deoxykana-mycin-B with a a-Hydroxy-co-amino fatty acid, and it was then found that the resulting new lN- [a-hydroxy-co-aminoalkanoyl] derivatives of 5,3 ', 4'-trideoxykanamycin-B, 5,3', 4 ' -Trideoxy-6'-N-methylkanamycin-B and 5.3 ', 4', 6 "-tetradeoxykanamycin-B have a very high activity against kanamycin sensitive and kanamycin resistant bacteria and also a high antibacterial activity against a wide range of bacteria develop.

s

The present invention thus relates to new compounds, namely 1N- [a-hydroxy-co-aminoalkanoyl] -5,3 ', 4'-trideoxy- or 1N- [a-hydroxy-co-aminoalkanoyl] -5,3', 4'-trideoxy-6'-N-methylkanamycin-B or 5,3 ', 4'-tri-i «deoxy-6'-N-methylkanamycin-B or an IN- [cc-hydroxy-co-aminoalkanoyl] -5 , 3 ', 4', 6 "-tetradeoxykanamycin-B or 5,3 ', 4', 6" -tetradeoxykanamycin-B, which correspond to the following formula I:

in which R represents a hydroxyl group or a hydrogen atom, 30 or a methyl group, provided that Ri represents a hydrogen atom or an a-hydroxy-co-aminoalka- that R2 is not the methyl group when R represents hydrogen, or a pharmaceutically acceptable acid addition salt thereof Links.

noyl group of the formula -CO-CH- (CH2) n-NH2

I.

OH

where n is 1, 2 or 3 and R2 is a hydrogen atom

6 "

These new compounds of formula I according to the invention include a new compound, namely 1N- [a-hydroxy-ra-aminoalkanoyl] -5,3 ', 4'-trideoxykanamycin-B or 1N- [a-hydroxy-co-aminoalkanoyl] -5,3 ', 4', 6 "-tetradeoxykanamycin-B of formula II

choh

(ch2) nnh2

[IIJ

in which R represents a hydroxyl group or a hydrogen atom and n represents 1, 2 or 3, and in which R represents the hydroxyl group if the compound of the formula II is an IN - [a-hydroxy- <a-aminoalkanoyl] -5,3 ', 4 '-trideoxykanamycin-B but R is hydrogen when the compound of formula II is an IN- [a-hydroxy-co-aminoalkanoyl] -5,3', 4 ', 6 "-

is tetradeoxykanamycin-B, and the pharmaceutically acceptable acid addition salts of the compounds of formula II.

Another preferred compound which belongs to the compounds of the formula I according to the invention is 5,3 ', 4', 6 "tetradeoxykanamycin B of the formula III

11

648564

[iii]

as well as their pharmaceutically acceptable acid addition salts.

The physico-chemical and biological properties of the compounds of formulas II and III are as follows:

(1) 1 -N - [(S) -4-amino-2-hydroxybutyryl] -5,3 ', 4' -trideoxy-kanamycin-B-monocarbonate monohydrate is a substance in the form of a colorless powder, which can be found in Decomposed 163 to 166 ° C and a specific optical rotation

[a] o = + 87 ° C (c = 1, water). Your elementary analysis agrees with the theoretical values of C :: H44Nr> Oj-H: COj-H2O (C 44.80%, H 7.85%, N 13.63%). This substance gives a single spot (positive for ninhydrin) at Rf 0.05 and at Rf 0.09 in a thin layer chromatography on silica gel, developed with butanol - ethanol - chloroform - 17% aqueous ammonia (in a volume ratio of 4: 5: 2: 5 ) or with chloroform - methanol - concentrated aqueous ammonia - water (volume ratio 1: 4: 2: 1)) as developer solvent.

(2) 1 -N - [(RS) -3-amino-2-hydroxypropionyl] -5,3 ', 4' -tri-deoxykanamycin-B-monocarbonate is a substance in the form of a colorless powder, which is found in 113 to 116 ° C decomposed and has a specific optical rotation [cc] d = + 120 ° (c = 1, water). Your elemental analysis agrees with the theoretical values of CmH ^ NeO * H2CO3 (C 42.85%, H 7.19%, N 13.63%). This substance gives a single spot (positive for ninhydrin) at Rf 0.12 and at Rf 0.35 in the above-mentioned thin layer chromatography on silica gel, developed with butanol - ethanol - chloroform - 17% aqueous ammonia (volume ratio 4: 5: 2: 5) or with chloroform - methanol - concentrated aqueous ammonia - water (volume ratio 1: 4: 2: 1) as developer solvent.

(3) IN - [(S) -4-amino-2-hydroxybutyryl] -5,3 ', 4', 6 "-tetra-deoxykanamycin-B-dicarbonate is a substance in the form of a colorless powder, which is found in 131 decomposed up to 135 ° C and a specific optical rotation [a] ?, = + 90 °

(c = 1, water). Your elementary analysis agrees with the theoretical values of C22H44N6O8 • 2H2CO3 (C

15 44.71%, H 7.50%, N 13.04%). This substance gives a single spot (positive for ninhydrin) at Rf 0.07 and at Rf 0.23 in the above-mentioned thin layer chromatography on silica gel, developed with butanol - ethanol - chloroform - 17% aqueous ammonia (volume ratio 20 4: 5: 2 : 5) or with chloroform - methanol - concentrated aqueous ammonia - water (volume ratio 1: 4: 2: 1) as developer solvent.

(4) 5,3 ', 4', 6 "-tetradeoxykanamycin-B-dicarbonate mono-25 hydrate is a substance in the form of a colorless powder which decomposes at 128 to 136 ° C and has a specific optical rotation [a]" = + 102 ° (c = 1, water). Your elementary analysis agrees with the theoretical values of CisHSTNSOÔ • 2H2CO3-H20 (C42.77%, H7.72%, N 12.47%) 30. This substance gives a single spot (positive for ninhydrin) at Rf 0.42 and at Rf 0.56 in the above-mentioned thin layer chromatography on silica gel, developed with butanol - ethanol - chloroform - 17% aqueous ammonia (volume ratio 4: 5: 2: 5) or with 35 chloroform - methanol - concentrated aqueous ammonia - water (volume ratio 1: 4: 2: 1) as developer solvent.

The minimal inhibitory concentration (MIC) (mcg / ml) 40 of IN - [(S) -4-amino-2-hydroxybutyryl] -5,3 ', 4'-trideoxyka-namycin-B (abbreviated as AHB-trideoxy- KMB), 1-N - [(RS) -3-amino-2-hydroxypropionyl] -5,3 ', 4' -trideoxykanamycin-B (abbreviated to AHP-trideoxy-KMB) and lN - [(S) -4 -Amino-2-hydroxybutyryl] -5,3 ', 4', 6 "-tetradeoxykanamycin-B (abge-45 abbreviated with AHB-tetradeoxy-KMB) of the formula II and the new compound 5,3 ', 4', 6 "-Tetradeoxykanamycin-B (abbreviated to tetradeoxy-KMB) of the formula III against various microorganisms was determined by serial dilution methods on a nutrient agar medium at 50 37 ° C. and assessment after 18 hours of incubation. For comparison purposes, the minimum inhibitory concentrations of habekacin were also determined in the same way.

The antibacterial spectra of these new and the 55 known substances are summarized in Table I below.

Table I

Test organisms

MIC (mcg / mI)

AHB trideoxy KMB

AHP trideoxy KMB

AHB tetradeoxy KMB

Habekacin (comparison)

Tetradeoxy KMB

Staphylococcus aureus 209P <0.20 <0.20 <0.20 0.39 3.13

Staphylococcus aureus Smith <0.20 <0.20 <0.20 <0.20 <0.20

Staphylococcus aureus AP01 0.39 0.78 0.78 0.78 6.25

Staphylococcus epidermidis 109 0.39 0.78 0.78 0.78 6.25

Micrococcus flavus FD A 16 1.56 6.25 6.25 1.56 50

648564

12

Table I (continued)

Test organisms

MIC (mcg / ml)

AHB trideoxy

AHP trideoxy

AHB tetradeoxy

Habekacin

Tetradeoxy

KMB

KMB

KMB

(Comparison)

Sarcina lutea PCI 1001

0.39

0.78

0.78

1.56

50

Bacillus anthracis

<0.20

<0.20

<0.20

<0.20

0.39

Bacillus subtilis PCI 219

<0.20

<0.20

<0.20

<0.20

<0.20

Bacillus subtilis NRRL B-558

<0.20

<0.20

<0.20

<0.20

<0.20

Bacillus cereus ATCC 10702

0.78

0.78

0.39

1.56

3.13

Mycobacterium smegmatis ATCC 607

0.20

0.20

<0.20

0.39

0.78

Escherichia coli NIHJ

0.78

1.56

0.78

3.13

6.25

Escherichia coli K-12

0.78

1.56

0.78

3.13

25th

Escherichia coli K-12 R5

100

100

50

100

> 100

Escherichia coli K-12 R 388

0.78

1.56

0.39

1.56

3.13

Escherichia coli K-12 J5R 11-2

0.78

1.56

0.78

1.56

6.25

Escherichia coli K-12 ML 1629

1.56

3.13

0.78

3.13

6.25

Escherichia coli K-12 ML 1630

1.56

3.13

1.56

3.13

12.5

Escherichia coli K-12 ML 1410

1.56

1.56

1.56

6.25

12.5

Escherichia coli K-12 ML 1410 R81

1.56

1.56

0.78

3.13

6.25

Escherichia coli K-12 LA 290 R55

3.13

3.13

1.56

6.25

> 100

Escherichia coli K-12 LA 290 R56

0.78

1.56

0.39

3.13

100

Escherichia coli K-12 LA 290 R64

0.78

1.56

0.78

3.13

100

Escheri chia coli W677

0.78

1.56

0.78

3.13

6.25

Escherichia coli JR66 / W677

1.56

3.13

1.56

6.25

> 100

Escherichia coli K-12 C 600R / 35

0.78

1.56

0.78

1.56

6.25

Escherichia coli JR255

0.39

1.56

0.78

3.13

25th

Klebsiella pneumoniae PCI 602

0.78

1.56

0.78

1.56

3.13

Klebsiella pneumoniae 22 # 3038

3.13

6.25

1.56

3.13

100

Shigella dysenteriae JS 11910

3.13

6.25

1.56

12.5

12.5

Shigella flexneri 4b JS 11811

3.13

6.25

1.56

6.25

25th

Shigella sonnei JS11746

3.13

6.25

3.13

6.25

12.5

Salmonella typhi T-63

25th

50

25th

1.56

12.5

Salmonella enteritidis 1891

3.13

6.25

3.13

3.13

25th

Proteus vulgaris OX19

0.39

0.78

0.39

0.78

1.56

Proteus rettgeri GN311

25th

25th

25th

50

> 100

Proteus rettgeri GN466

3.13

3.13

1.56

12.5

"25

Serratia marcescens

25th

25th

25th

50

100

Serratia SOU

100

> 100

100

> 100

> 100

Serratia 4

12.5

25th

25th

50

50

Providencia Pv 16

12.5

25th

25th

25th

<100

Providencia 2991

6.25

25th

25th

50

<100

Pseudomonas aeruginosa A3

0.39

0.78

0.78

3.13

3.13

Pseudomonas aeruginosa No.12

3.13

3.13

6.25

3.13

25th

Pseudomonas aeruginosa H9

3.13

3.13

3.13

. 6.25

100

Pseudomonas aeruginosa Hl 1

6.25

6.25

12.5

12.5

25th

Pseudomonas aeruginosa TI-13

3.13

3.13

3.13

3.13

12.5

Pseudomonas aeruginosa GN315

25th

50

100

6.25

> 100

Pseudomonas aeruginosa 99

3.13

3.13

6.25

6.25

25th

Pseudomonas aeruginosa B-13

6.25

12.5

12.5

25th

50

Pseudomonas aeruginosa 21-75

6.25

12.5

12.5

25th

> 100

Pseudomonas aeruginosa PST1

6.25

6.25

6.25

25th

> 100

Pseudomonas aeruginosa

> 100

> 100

100

> 100

> 100

ROS134 / PU21

Pseudomonas aeruginosa K-Psl02

1.56

3.13

3.13

3.13

12.5

Pseudomonas maltophilia GN907

> 100

> 100

> 100

> 100

> 100

Another preferred new compound, which belongs to the co-aminoalkanoyl] -5,3 ', 4'-trideoxy-6'-N-methyl-channel compounds of the formula I, is a 1 -N- [a-hydroxy-mycin- B of formula IV

13

648564

hn i 1

co choh i

(ch2) nnh2

[iv]

in which n is 1, 2 or 3, and their pharmaceutical compounds belong to the formula I, is 5,3 ', 4'-trideoxy-6' acceptable acid addition salts. N-methylkanamycin-B of formula V

Another interesting new compound which is related to the and their pharmaceutically acceptable acid addition salts.

The physico-chemical and biological properties of the above-mentioned compounds of formulas IV and V are as follows:

(5) 1 -N - [(S) -4-amino-2-hydroxybutyryl] -5,3 ', 4'-trideoxy-6'-N-methylkanamycin-B-monocarbonate is a substance in the form of a colorless powder, which decomposes at 162 to 165 ° C and has a specific optical rotation [a] o = + 88 ° (c = 1, water). Your elementary analysis agrees with the theoretical values for C23H46N6O9 • H2CO3 (C 47.05%, H 7.90%, N 13.72%). This substance gives a single spot (positive for ninhydrin) at Rf 0.05 and at Rf 0.08 in a thin layer chromatography on silica gel, developed with butanol - ethanol - chloroform - 17% aqueous ammonia (volume ratio 4: 5: 2: 5) or with chloroform - methanol - concentrated aqueous ammonia - water (volume ratio 1: 4: 2: 1) as developer solvent.

(6) 1 -N - [(RS) -3-amino-2-hydroxypropionyl] -5,3 ', 4'-tri-deoxy-6'-N-methylkanamycin-B-monocarbonate monohydrate is a substance in the form of a colorless powder,

which decomposes at 162 to 164 ° C and has a specific optical rotation [a] n = + 80 ° (c = 0.5, water). Your elemental analysis agrees with the theoretical values of C :: H44N6Û9- H: C03- H: 0 (C44.80%, H7.85%, N13.62%). This substance gives a single stain (positive

40 on ninhydrin) at Rf 0.05 and at Rf 0.14 in the above-mentioned thin layer chromatography on silica gel, developed with butanol - ethanol - chloroform - 17% aqueous ammonia (volume ratio 4: 5: 2: 5), or with chloroform - methanol - concentrated aqueous ammo-

4s niac - water (volume ratio 1: 4: 2: 1) as developer solvent.

(7) 1 -N - [(S) -5-amino-2-hydroxyvaleryl] -5,3 ', 4'-trideoxy-6' -N-methylkanamycin-B-monocarbonate monohydrate is a substance in the form of a colorless Powder, which itself

50 decomposed at 163 to 166 ° C and has a specific optical rotation [a] o = + 84 ° (c = 0.5, water). Your elementary analysis agrees with the theoretical values of C24H48N6O9 • H2CO3 • H20 (C46.57%, H8.13%, N 13.03%).

55

This substance gives a single stain (ninhydrin positive) at RF 0.03 and RF 0.08 in the thin layer chromatography on silica gel developed above with butanol-ethanol-chloroform-17% aqueous ammonia

60 (volume ratio 4: 5: 2: 5) or with chloroform-methanol-concentrated aqueous ammonia water (volume ratio 1: 4: 2: 1) as developer solvent.

(8) 5,3 ', 4' -trideoxy-6 '-N-methylkanamycin-B-monocar bonate monohydrate is a substance in the form of a colorless

65 powder, which decomposes at 137 to 140 ° C and has a specific optical rotation jcxJjj = + 66 ° (c = 1, water). Your elementary analysis agrees with the theoretical values of C19H39N5OT H2C03- H20 (C45.36%,

648564

14

H 8.18%, N 13.23%). This substance gives a single spot (positive for ninhydrin) at Rf 0.22 and at Rf 0.49 in the above-mentioned thin layer chromatography on silica gel, developed with butanol - ethanol - chloroform - 17% aqueous ammonia (volume ratio 4: 5: 2: 5) or with chloroform - methanol - concentrated aqueous ammonia - water (volume ratio 1: 4: 2: 1) as developer solvent.

The minimum inhibitory concentrations (MIC) (mcg / ml) of IN - [(S) -4-amino-2-hydroxybutyryl] -5,3 ', 4'-trideoxy-6'-N-methylkanamycin-B (abbreviated as AHB-Tri-deoxy-MKMB), 1 -N - [(RS) -3-Amino-2-hydroxypropionyl] -5,3 ', 4'-trideoxy-6'-N-methylkanamycin-B (abbreviated AHP- Tridoeoxy-MKMB) and IN - [(S) -5-Amino-2-hydroxy-

vaIeryl] -5,3 ', 4'-trideoxy-6'-N-methylkanamycin-B (abbreviated to AHV-Trideoxy-MKMB) of the formula IV and the compound 5,3', 4'-trideoxy-6'-N -methylkanamycin-B (abbreviated to trideoxy-MKMB) of the formula V against various microorganisms was determined using the serial dilution method on a nutrient agar medium at 37 ° C., the evaluation being carried out after 18 hours of incubation. For comparison purposes, the minimum inhibitory concentration of l-N - [(S) -4-amino-2-hydroxy-10-butyryl-3 ', 4'-dideoxykanamycin-B (i.e. habekacin) was also determined in the same manner as above.

The antibacterial spectra of these new and known compounds are summarized in Table II below.

Table II

Test organisms

MIC (mcg / ml)

AHB-trideoxy-MKMB

AHP-trideoxy-MKMB

AHV-trideoxy-MK.MB

Habekacin (comparison)

Trideoxy MKMB

Staphylococcus aureus 209P

<0.20

0.39

0.20

0.39

1.56

Staphylococcus aureus Smith

<0.20

<0.20

<0.20

<0.20

0.39

Staphylococcus aureus AP01

0.39

<0.20

0.39

0.78

25th

Staphylococcus epidermidis 109

<0.20

<0.20

<0.20

0.78

6.25

Micrococcus flavus FDA16

1.56

3.13

3.13

1.56

100

Sarcina lutea PCI 1001

0.39

0.78

1.56

1.56

25th

Bacillus anthracis

<0.20

<0.20

<0.20

<0.20

0.78

Bacillus subtilis PCI 219

<0.20

0.39

<0.20

<0.20

<0.20

Bacillus subtilis NRRL B-558

<0.20

0.39

<0.20

<0.20

0.78

Bacillus cereus ATCC 10702

0.39

1.56

1.56

1.56

12.5

Mycobacterium smegmatis ATCC 607

<0.20

0.39

0.39

0.39

6.25

Escherichia coli NIHJ

0.78

3.13

1.56

3.13

12.5

Escherichia coli K-12

0.78

3.13

1.56

3.13

12.5

Escherichia coli K-12 R5

3.13

12.5

6.25

100

50

Escherichia coli K-12 R 388

0.39

1.56

1.56

1.56

12.5

Escherichia coli K-12 J5R 11-2

1.56

3.13

3.13

1.56

25th

Escherichia coli K-12 ML 1629

0.78

3.13

3.13

3.13

25th

Escherichia coli K-12 ML 1630

1.56

3.13

3.13

3.13

25th

Escherichia coli K-12 ML 1410

1.56

3.13

6.25

3.13

25th

Escherichia coli K-12 ML 1410 R81

1.56

3.13

3.13

3.13

25th

Escherichia coli K-12 LA 290 R55

1.56

6.25

3.13

6.25

100

Escherichia coli K-12 LA 290 R56

0.78

3.13

1.56

3.13

25th

Escherichia coli K-12 LA 290 R64

1.56

3.13

1.56

3.13

25th

Escherichia coli W677

1.56

3.13

1.56

3.13

12.5

Escherichia coli JR66 / W677

1.56

6.25

3.13

6.25

100

Escherichia coli K-12 C 600R / 35

1.56

1.56

1.56

1.56

12.5

Escherichia coli JR255

0.78

3.13

1.56

3.13

100

Klebsiella pneumoniae PCI 602

0.78

3.13

1.56

1.56

12.5

Klebsiella pneumoniae 22 # 3038

1.56

3.13

3.13

3.13

100

Shigella dysenteriae JS 11910

1.56

6.25

6.25

12.5

50

Shigella flexneri 4b JS 11811

0.78

3.13

1.56

6.25

25th

Shigella sonnei JS11746

3.13

6.25

6.25

6.25

25th

Salmonella typhi T-63

1.56

3.13

3.13

1.56

6.25

Salmonella enteritidis 1891

0.78

6.25

3.13

3.13

25th

Proteus vulgaris OX19

<0.20

0.78

0.78

0.78

3.13

Proteus rettgeri GN311

50

25th

100

50

100

Proteus rettgeri GN466

6.25

12.5

6.25

12.5

25th

Serratia marcescens

6.25

12.5

12.5

50

100

Serratia SOU

25th

50

50

> 100

> 100

Serratia 4

6.25

12.5

12.5

50

50

Providencia Pv 16

6.25

50

25th

25th

> 100

Providencia 2991

12.5

50

25th

50

> 100

Pseudomonas aeruginosa A3

0.69

0.78

0.39

3.13

3.13

Pseudomonas aeruginosa No. 12

1.56

3.13

3.13

3.13

25th

Pseudomonas aeruginosa H9

1.56

3.13

3.13

6.25

25th

Pseudomonas aeruginosa Hl 1

12.5

12.5

6.25

12.5

25th

Pseudomonas aeruginosa TI-13

1.56

3.13

1.56

3.13

12.5

Pseudomonas aeruginosa GN315

3.13

3.13

3.13

6.25

25th

15

Table II (continued)

648564

Test organisms MIC (mcg / ml)

AHB-trideoxy- AHP-trideoxy- AHV-trideoxy- Habekacin trideoxy-MKMB

MKMB MKMB MKMB (comparison)

Pseudomonas aeruginosa 99

3.13

12.5

6.25

6.25

50

Pseudomonas aeruginosa B-13

12.5

25th

12.5

25th

100

Pseudomonas aeruginosa 21-75

12.5

25th

12.5

25th

> 100

Pseudomonas aeruginosa PST1

12.5

25th

12.5

25th

> 100

Pseudomonas aerugionsa

50

50

50

> 100

> 100

ROS134 / PU21

Pseudomonas aeruginosa K-Psl02

1.56

3.13

1.56

3.13

25th

Pseudomonas maltophilia GN907

> 100

> 100

> 100

> 100

> 100

It can be seen from Tables I and II that the new compounds of the formula I according to the invention, including the compounds of the formulas II, III, IV and V, prevent the growth of numerous types of bacterial strains. The new compounds of the present invention have low acute toxicity to animals and humans. It has been found that the new compounds of formulas II and III, e.g. 1-N - [(S) -4-amino-2-hydroxybu-tyryl] -5,3 ', 4'-trideoxykanamycin-B; 1-N- [3-Amino-2-hydro-xypropionyl] -5,3 ', 4'-trideoxykanamycin-B; IN - [(S) -4-amino-2-hydroxybutyryl] -5,3 ', 4', 6 "-tetradeoxykan-mycin-B and 5,3 ', 4', 6" -tetradeoxykanamycin-B an LDso- Have a value of 25 to 50 mg / kg if their acute toxicity is determined by intravenous injection in mice. It has also been found that the new compounds of formulas III and IV, e.g. l-N - [(S) -4-amino-2-hydroxybu-tyryl] -5,3 ', 4'-trideoxy-6'-N-methylkanamycin-B; l-N- (3-amino-2-hydroxypropionyl) -5,3 ', 4' -trideoxy-6 '-N-methyl-kanamycin-B; 1 -N - [(S) -5-amino-2-hydroxyvaleryl] -5,3 ', 4' -trideoxy-6'-N-methylkanamycin-B and 5,3 ', 4'-trideoxy-6' -N-methylkanamycin-B have an LD 50 of 50 to 100 mg / kg if their acute toxicity is determined by intravenous injection in mice.

The new compounds according to the invention are usually obtained in the form of their free base or a hydrate or a carbonate thereof. The new compounds of the invention can each easily be converted into the form of a pharmaceutically acceptable acid addition salt, e.g. into the hydrochloride, hydrobromide, sulfate, phosphate, nitrate, acetate, maleate, citrate, ascorbate, methanesulfonate and the like, by reaction with the corresponding pharmaceutically acceptable inorganic or organic acid in an aqueous medium.

The new compounds of formulas I, II, III, IV or V and their pharmaceutically acceptable acid addition salts can be administered orally, interperitoneally, intravenously, subcutaneously or intramuscularly using any known pharmaceutical form for such administration and in a manner similar to the known Kana -mycine. For example, the new compounds can be administered orally using any known pharmaceutical form for oral administration. Examples of pharmaceutical forms for oral administration are powders, capsules, tablets, syrups and the like. A suitable dose for the novel compounds according to the invention for achieving an effective treatment of bacterial infections is in the range from 0.1 to 1 g per person per day when administered orally. This dose should preferably be administered orally in three to four aliquots per day. The new compounds can also be administered by intramuscular injection at a dose of 50 to 500 mg per person two to four times a day. In addition, the new compounds can also be processed into an ointment for external use, which contains the active compound in a concentration of 0.5 2s to 5% by weight in a mixture with a known ointment base, e.g. Polyethylene glycol, contains. Furthermore, the new compounds are all also useful for the sterilization of surgical instruments and sanitary materials.

Another object of the present invention is therefore an antibacterial preparation which, as an active ingredient, an IN- [a-hydroxy-co-aminoalkanoyl] -5,3 ', 4'-trideoxykanamycin-B, an IN- [a-hydroxy- co-aminoa! ka-noyl] -5,3 ', 4', 6 "-tetradeoxykanamycin-B or a pharmaceutically acceptable acid addition salt thereof, as represented by 35 Formula II, or 5,3 ', 4', 6" - Tetradeoxykana-mycin-B or a pharmaceutically acceptable acid addition salt thereof, as represented by the formula III, in an antibacterially effective amount to prevent the growth of bacteria, together with a carrier for the 4o active compound, and an antibacterial preparation which as an active ingredient, an IN- [a-hydroxy-co-aminoalkanoyl] -5,3 ', 4'-trideoxy-6'-N-methylkanamycin-B or a pharmaceutically acceptable acid addition salt thereof, as represented by the formula IV, or 5 , 3 ', 4'-Tri-45 deoxy-6'-N-methylkanamycin-B or a pharmaceutically acceptable acid addition salt thereof, as represented by the formula V, in an antibacterially effective amount to prevent the growth of bacteria, together with a carrier for the active compound.

Next, the preparation of the new compounds of formula II will be described. The new compounds of formula II, e.g. IN - [(S) -4-amino-2-hydroxybutyryl] -5,3 ', 4'-trideoxykanamycin-B and IN - [(S) -4-amino-2-hydroxybu-55 tyryl] -5,3 ', 4', 6 "-tetradeoxykanamycin-B can be prepared using either the known compound 5,3 ', 4'-trideoxykanamycin-B (Japanese Journal of Antibiotics, 32, S-178 [1979]) or the new one Compound 5,3 ', 4', 6 "-tetradeoxykanamycin-B, as it is obtained according to the present invention, as starting substance and condensing the 1-amino group of the starting compound with a corresponding a-hydroxy-co-amino fatty acid or functional equivalent of it.

Another object of the present invention is accordingly a process for the preparation of an IN- [a-hydroxy-co-aminoalkanoyi] -5,3 ', 4'-trideoxykanamycin-B or 1 -N- [a-hydroxy-co-aminoalkanoyl ] -5,3 ', 4', 6 "-tetradeoxykanamycin-B according to formula II

648564

16

hn i

co choh

[ii]

(ch2) nnh2

in which R is a hydroxyl group or a hydrogen atom (a) is the 1-amino group of 5,3 ', 4'-trideoxykanamycin-B and n is 1, 2 or 3, which is characterized or 5,3', 4 ', 6 " -Tetradeoxykanamycin-B of formula VI is that one

[vi]

in which R has the same meaning as above, or a partially amino-protected derivative of 5,3 ', 4'-trideoxykanamycin-B or 5,3', 4 ', 6' formula VI '

-Tetradeoxykanamycin-B the n '

-a

-B

[vi ']

in which R is a hydroxyl group or a hydrogen atom and A is a hydrogen atom and at least one B is a monovalent amino protective group, but the other B are each a hydrogen atom or at least one pair A and B together are a divalent amino protective group, but the other A and B are each a hydrogen atom, wherein the amino protecting groups represented by A and B may be the same or different, by reaction with an a-hydroxy-co-amino fatty acid or an amino-protected derivative thereof of the formula VII

60 ^ "N (CH2) nCHCOOH (VII)

B '^ |

OH

in which A 'is a hydrogen atom and B' is a hydrogen atom or a monovalent amino protective group or A '6s and B' together form a divalent amino protective group and n is 1, 2 or 3, or a functional equivalent of the compound of the formula VII, in order to -N-acylated product of the formula II '

17th

648564

[ii ']

or formula II "

choh

(ch2) nn b1

[ii "]

in which R, A, B, A ', B' and n have the same meaning as above, and

(b) the remaining amino protecting groups, if any, are removed from the 1-N-acylated product of formula II 'or II "in a known manner to obtain the compound of formula II.

The process according to the invention may comprise a further step in which the compound of formula II is converted into a pharmaceutically acceptable acid addition salt by reaction with a pharmaceutically acceptable inorganic or organic acid in a known manner, if desired.

The procedure for carrying out the method according to the invention will now be described in detail. In carrying out the present process, it is possible to use 5,3 ', 4'-trideoxykanamycin-B or 5,3', 4 ', 6 "-tetradeoxykanamycin-B (VI) as starting compound, in which the amino groups are not protected at all These starting compounds can be present in the form of the free base or the acid addition salt with a suitable acid, for example hydrochloric acid or sulfuric acid, but a partially amino-protected derivative of 5,3 ', 4'-trideoxykana-mycin-B is preferred as the starting compound or 5,3 ', 4', 6 "-tetradeoxykanamycin-B of formula VI ', in which all or part of the amino groups with the exception of the 1-amino group were protected with known amino protective groups and 4s which by introducing a known amino protective group into the Compound of formula VI can be prepared in a known manner as used in the synthesis of some known deoxy derivatives of kanamycin-B. For the production of partially amino-protected 5,3 ', 4'-so trideoxykanamycin-B or 5,3', 4 ', 6 "-tetradeoxykan-mycin-B of the formula VI', the amino protection methods which are used in the Preparation of 6'-N-benzyloxycarbonyl derivatives of Kanamycin-B as described in U.S. Patent No. 3,781,168 or No. 3,929,762; ss in the preparation of 2 ', 6'-di-N-tert-butoxycarbonylka -mamycin-B or 6'-N-benzyloxycarbonylkanamycin-B or the mono-N- or di-N-tert-butoxycarbonyl and even tri-N-tert-butoxycarbonyl derivative of 6'-N-benzyloxycar-bonyl-kanamycin -B either isolated or in a mixture as described in British Patent No. 1,426,908 or US Patent No. 3,939,143; or in the preparation of 2 ', 3.3 ", 6'-tetra-N-formyl derivatives of Kanamycin-B, as in Belgian Patent No. 817 546 can be used.

65 In general, suitable examples of amino protective groups which can be used for the protection of some amino groups in the partially amino-protected derivative of the formula VI 'are a common amino protective group,

648564

18th

including an alkoxycarbonyl group such as tert-butoxoxycarbonyl and tert-amyloxycarbonyl; a cycloalkyloxy carbonyl group such as cyclohexyloxycarbonyl; an aralkyloxycarbonyl group such as benzyloxycarbonyl; an acyl group such as trifluoroacetyl and o-nitrophenoxyacetyl; a s phosphinothioyl group such as diphenylphosphinothioyl and dimethylphosphinothioyl; a phosphinyl group such as diphenylphosphinyl and the like. Preferred examples of divalent amino protecting groups include the phthaloyl group and a Schiff see 10 group, such as salicylidene. The amino protective group of this type can be introduced by reacting the compound of the form] VI with a suitable known reagent for introducing the amino protective group, which can be in the form of an acid halide, acid azide, active ester or acid anhydride and the like, as is the case with the conventional one Synthesis of peptides is known. By selecting the amount of the reagent used to introduce the amino protecting group in a ratio of 0.5 to 6 moles per mole of compound of formula VI, it is possible to obtain a mixture of various partially amino protected derivatives (VI ') in any ratio as a result the different reactivity of the corresponding amino groups of the compound of formula VI.

In the present process, it is advisable to use such an amino-protected 5,3 ', 4'-trideoxy-kanamycin-B or 5,3', 4 ', 6 "-tetradeoxykanamycin-B derivative as the starting compound, in which all or some of the amino groups with the exception of the 1-amino group have been blocked, for example a 3,2 ', 6', 3 "tetra-N-protected derivative, a 30 3,2 ', 6'-tri-N-protected derivative , a 2 ', 6', 3 "-tri-N-protected derivative, a 2 ', 6', 3" -tri-N-protected derivative, a 2 ', 6'-di-N-protected derivative and a 6'-mono-N protected derivative. Furthermore, a mixture of two or more of these partially N-protected derivatives can be used without purification 35 for the 1-N-acylation in the present process.

In order to ensure that the desired product of the general formula II can be produced in high yield by the process according to the invention, it is only necessary that only the 1-amino group of the compound of the formula VI, namely 5,3 ', 4' -Trideoxykanamycin-B or 5,3 ', 4', 6 "-Tetradeoxykanamycin-B, is selectively aeylated with the a-hydroxy-co-amino fatty acid (VII). It follows that preferably a 3,2 ', 6' , 3 "tetra-N-45

protected derivative of compound VI, i.e. the amino-protected derivative of the compound of the formula VI ', in which all amino groups with the exception of the 1-amino group have been blocked with protective groups, is used as the starting compound for 1-N-acylation in the present process.

To prepare the 3,2 ', 6', 3 "tetra-N-protected derivative of formula VI 'from the compound of formula VI, for example, the following procedure can be used. A known method according to US Pat. 4 136 254 of 55

Nagabhushan et al. can be used in which a 3,2 ', 6'-tri-N-acylated protected derivative of kanamycin-B is prepared by reacting kana-mycin-B with a divalent transition metal cation, e.g. a cation of copper (II), nickel (II), cobalt (II), etc. 60 for the formation of a metal complex of kanamycin-B, reaction of this kanamycin B-metal complex with an acylating agent which is known as an agent for introducing amino protective groups to protect all amino groups except the 1 - amino and 3 "amino groups of the channel 65 mycin B portion of the kanamycin B metal complex, which 1- and 3" amino groups by complexing with the divalent metal cation in Canada

mycin-B metal complex was blocked, and then removing the divalent metal cations from this complex, e.g. by treatment with hydrogen sulfide or with aqueous ammonia. Or a method can be used as described in Belgian Patent No. No. 879,925 of the patent owner, in which a 3,2 ', 6'-tri-N-acylated, protected derivative of kanamycin-B is prepared, in a similar manner to that described by the Nagabhushan et al Zinc cations are used instead of the divalent transition metal cations. In this way, a 3,2 ', 6' -tri-N-protected derivative of the formula VI 'can be prepared from the compound of the formula VI in high yield. The 3 "amino group of the 3,2 ', 6' -tri-N-protected derivative (VI ') thus obtained can be further protected by selective acylation according to a selective 3" -N-acylation method as described in claim 15 of the Belgian Patentes No. 879 923 to obtain an amino-protected derivative of an aminoglycosidic antibiotic, in which all amino groups with the exception of the 1-amino group were selectively protected, so that a 3,2 ', 6', 3 "-tetra-N-protected derivative of Compound VI can be produced in high yield. According to the selective 3 "-N-acylation method as described in claim 15 of Belgian patent no. 879,923, the above-mentioned 3,2 ', 6'-tri-N-protected derivative of the compound VI is reacted with an alkyl formate, an alkyl dihalogen or trihalogenate, formylimidazole or an N-alkanoylimidazole as acylating agent, whereby the 3 "-amino group can be selectively aeylated with the acyl residue of the acylating agent used in high yield without the acylation of the 1-amino group of the 3,2 ', 6'-tri-N-protected derivative occurring. The 3,2' , 6 ', 3 "-tetra-N-acylated derivative, e.g. 3,2 ', 6'-tri-N -benzyloxycarbonyl-3 "-N-trifluoroacetyl derivative of 5,3', 4 'tri-deoxykanamycin-B or 5,3', 4 ', 6" tetradeoxykana- mycin-B, which by using the above-mentioned methods according to US Patent No. 4 136 254 and Belgian patent no. 879 923 can be obtained is a preferred starting material for the selective 1-N-acylation with the a-hydroxy-co-amino fatty acid (VII) in the 1-N-acylation stage of the present process.

In this process, the 1-amino group of the compound of formula VI or the 1-amino group of partially amino-protected derivatives VI 'thereof, either isolated or in a mixture of two or more thereof, with the a-hydroxy-co-amino fatty acid of formula VII

; N- (CH2) "- CHC00H

I.

OH

(VII)

aeylated, in which A 'and B' mean the same as above and n represents 1, 2 or 3, and in which the amino group is neither protected nor protected. This a-hydroxy-co-amino fatty acid can be 3-amino-2-hydroxypropionic acid (ie the compound of formula VII in which n = 1 and A 'and B' are hydrogen), 4-amino-2-hydroxybutyric acid (ie the compound of formula VII in which n = 2 and A 'and B' are hydrogen) or 5-amino-2-hydroxyvaleric acid (ie the compound of formula VII in which n = 3 and A 'and B' are hydrogen) be. Among them, the (S) isomer is preferred.

In the process according to the invention, 1-N-acylation with the a-hydroxy-co-amino fatty acid (VII) can be carried out by any customary method for the synthesis of peptides, for example by the known dicyclohexylcarbodi-

19th

648564

imide method, the known mixed acid anhydride method, the known azide method or the active ester method and the like, using a-hydroxy-co-amino fatty acid as such or in the form of its reactive derivatives (as a functional equivalent thereof). For the amino protecting group for protecting the amino group of the a-hydroxy-co-amino fatty acid, such an amino protecting group can be used which is the same or different from that in the starting compound VI '. A preferred amino protecting group for this purpose is in particular the tertiary butoxycarbonyl group, which is easily removable by treatment with aqueous trifluoroacetic acid or acetic acid or with dilute aqueous hydrochloric acid. The benzyloxycarbonyl group, which can be removed by conventional hydrogenolysis in the presence of a catalyst such as palladium or platinum oxide, forms a convenient N-protecting group.

The 1-N-acylation in the present process can preferably be carried out in an aqueous organic solvent, according to the active ester method and using an a-hydroxy-co-amino fatty acid in the form of its active ester. For example, the N-hydro-xysuccinimide ester of L-4-benzyloxycarbonylamino-2-hydroxybutyric acid can preferably be used as the active ester, which can be prepared by known methods for the preparation of the active esters. This active ester can preferably be used in an amount of 0.5 to 3 molar equivalents and preferably of 1 to 1.5 molar equivalents per mol of the starting compound VI or VI 'to be acylated. The aqueous organic solvent used in the reaction medium can be a water-miscible organic solvent, e.g. Dioxane, 1,2-dimethoxyethane, dimethylformamide, tetrahydrofuran and the like, mixed with water. The 1-N acylation can be carried out at room temperature or, if desired, at an elevated temperature of 20 to 90 ° C. and for a reaction time of several hours, preferably 5 to 6 hours.

When the 1-N-acylation in the present process is carried out using a partially amino-protected derivative as the starting compound, in which some, but not all, of the amino groups except the 1-amino group are protected, e.g. using the 6'-N-protected derivative of the starting compound VI, the acylation products obtained can be partially purified by column chromatography, e.g. on silica gel, so that the unreacted starting material is removed, whereby a mixture of the desired 1-N-mono-acylated product with the otherwise N-cylated products is obtained, as is the case in the synthesis of habekacin, namely lN- [(S) -4-amino-2-hydroxybutyryl] -

3 ', 4'-dideoxykanamycin-B is as described in U.S. Patent No. 4,107,424. These mixed acylation products can, without being purified and / or isolated, be immediately subjected to the subsequent step of the present process in which the protecting group is removed, followed by purification and isolation so that the desired 1-N-mono- acylated product is obtained.

In the second stage of the process io according to the invention, the 1-N-acylated product obtained in the 1-N-acylation stage (including the mixed acylation products) is subjected to the removal of the amino protective groups, provided that these are still present in the 1-N-acylation product. The protecting groups are split off in the usual way. Thus, the alkoxycarbonyl type amino protecting group can be obtained by weak acid hydrolysis with an aqueous solution of trifluoroacetic acid or acetic acid and the like or with a dilute aqueous solution of an inorganic acid, e.g. Hydrochloric acid, 20 can be performed. The aralkyloxycarbonyl group, e.g. the benzyloxycarbonyl group, can be split off by conventional catalytic reduction (hydrogenolysis).

If a phthaloyl group is present as an amino protecting group, it can be removed by heating in a solution of hydrazine hydrate in a lower alkanol.

The deprotected acylation product obtained in the second step of the present process may contain the desired 1-N-acylation product of Formula II along with isomers thereof. The desired 1-N- (a-hydroxy-ro-aminoalkanoyl) derivative II can be isolated chromatographically and purified using a cation exchanger containing carboxylic acid functions, e.g. "Amberlite CG-50" (a product from Rohm & Haas Co., USA) or "CM-Sephadex 33 C-25" (a product from Pharmacia Co., Sweden), the antibacterial activity of the eluate fractions using a suitable kanamycin-sensitive Strain and kanamycin-resistant strain of bacteria can be determined.

40

The 5,3 ', 4', 6 "-tetradeoxykanamycin-B, which is used as the starting compound in the process for the preparation of IN (a-hydroxy-co-aminoalkanoyl) -5,3 ', 4', 6" -tetradeoxykan-mycin -B is used according to the present method, 45 can be obtained starting from 3 ', 4', 6 "-trideoxyka-namycin-B, which has already been synthesized (this compound was described as 6" -deoxydibekacin in the description of Japanese Patent Application Laid-Open No 119 323/79 or British Offenlegungsschrift No. GB 2 058 774 A) or starting from an N, 0-protected derivative thereof of the formula VIII '

, A

[viii1]

in which A is a hydrogen atom and B is a monovalent ami-6s. Another object of the present invention is no-protecting group, or A and B together are a divalent protecting process for the preparation of 5,3 ', 4', 6 "-tetradeoxyka amino-protecting group and a the hydroxyl-namycin-B, as described above, which has the formula III

group protecting acyl group means. having:

648564

20th

NH2

[III]

which is characterized in that 2 "-0-protected derivatives of 3 ', 4', 6" -trideoxykana-

(a) the 4 "hydroxyl group of a penta-N protected and is mycin-B of Formula VIII

[viii]

in which A is a hydrogen atom and B is a monovalent amino-protecting acyl group of the same type as the no-protecting group or A and B together are a divalent amino-protecting group and D is a hydroxyl-protecting acyl group with a monovalent hydroxyl-

Hydroxyl protecting group D in the 2 "position of compound VIII protects to form the 4" -0-protected compound of formula VIII ':

n

■ A b-

[viii1]

in which A, B and D have the same meaning as above,

(b) reacting the 4 "-0-protected compound of formula VIII 'obtained with sulfuryl chloride in order to replace the 5-hydroxyl group with a chlorine atom and the corresponding

To form 5-chloro derivative

(c) the 5-chloro derivative obtained is reduced to remove the 5-chloro group and to obtain a protected derivative of 5,3 ', 4', 6 "tetradeoxykanamycin B of the formula IX:

[IX]

21st

648564

in which A, B and D have the same meaning as above, and

(d) cleaving the remaining hydroxyl protecting groups and amino protecting groups from the compound of formula IX in a known manner to obtain the compound of formula III.

In this method, the amino protecting groups present in penta-N-protected and 2 "-O-protected 3 ', 4', 6" -T rideoxykanamycin-B of formula VIII which are used as the starting material can be the same as those in the starting compound of the Formula VI 'may be present, which are used in the first inventive method described above. The preparation of the starting compound of formula VIII is described below.

In the first stage of the process, the 4 "hydroxyl group of the starting compound VIII is protected with a monovalent hydroxyl protecting group of the acyl type, which can be a lower alkanoyl group such as acetyl or an aroyl group such as benzoyl. The introduction of a such acyl group protecting the hydroxyl group in the 4 "hydroxyl group of the starting compound VIII is easily accomplished by reacting the starting compound VIII with a corresponding acylating reagent in the form of an acid anhydride, acid halide or active ester in a known manner, for example in an organic solvent such as pyridine, at a temperature from 10 to 50 ° C, preferably at room temperature. Preferred acylating reagents for this purpose are acetyl chloride or benzoyl chloride. In this acylation reaction, the 5-hydroxyl group of the starting compound VIII is hardly aylated by the acylation reagent due to the lower reactivity of the 5-hydroxyl group.

In this way the 1, 3,2 ', 6', 3 "-penta-N-protected and 2", 4 "-di-0-protected derivative and 3 ', 4', 6" -trideoxyka-namycin- B of formula VIII 'generated.

In the second stage of the process, the protected derivative of formula VIII 'thus obtained is subjected to deoxygenation on the 5-hydroxyl group in a known manner, as described in the Bulletin of the Chemical Society of Japan, vol. 51, page 2354 (1978). Thus, the protected derivative VIII 'is mixed with one to five times the molar amount of sulfuryl chloride in an organic solvent, e.g. dry pyridine, at a temperature below room temperature during

Reacted for 2 to 20 hours while stirring, whereby the 5-chloro derivative is obtained.

In the third stage of this process, the 5-chloro derivative obtained in this way is reduced in order to carry out the halogen elimination s. This removal of the 5-chloro group can be accomplished by reaction with a metal hydride, e.g. Tributyltin hydride, as described in the above-mentioned publication, or by conventional catalytic hydrogenation in the presence of Raney nickel. In this way, the N, 0-protected derivative of 5,3 ', 4', 6 "-tetradeoxykan-mycin-B of the formula IX is obtained.

In the fourth stage, the N, 0-protected 5,3 ', 4', 6 "tetradeoxykanamycin B derivative IX is freed from the protective groups. The monovalent hydroxyl-protecting is acyl groups (D), which in the 5-deoxygenated Compound IX present can be easily removed by alkaline hydrolysis at room temperature, e.g.

by dissolving compound IX in ammoniacal methanol (i.e. a mixture of aqueous ammonia and methanol). The amino protecting group present in the starting compound VIII is of the aralkyloxycarbonyl type, and amino protecting groups of this kind can be simultaneously removed by the catalytic hydrogenation to which the 5-chloro derivative is subjected in the third step of the process mentioned above. Amino protecting groups other than the aralkyloxycarbonyl group can be easily removed in a known manner, e.g. by hydrolysis with a weak acid. If the amino protecting group is a lower alkoxycarbonyl group, e.g. Ethoxycarbonyl, 30 it can be removed by alkaline hydrolysis with barium hydroxide.

It is also possible to carry out this latter process in a modified embodiment, in which 3 ', 4', 6 "-trideoxykanamycin-B is used as starting material, its 5-amino groups and subsequently its 2" and 4 "hydroxyl groups are protected with the same monovalent hydroxyl protecting groups (D) to form the N, 0-protected derivative of the formula VIII '.

40 The preparation of the penta-N-protected and 2 "-0-protected derivative of 3 ', 4', 6" -trideoxykanamycin-B of the formula VIII, which is used as the starting compound for this second process according to the invention, can be carried out as in the British Patent application No. GB 2 058 774 45. So is a penta-N protected derivative of 3 ', 4'-dideoxykanamycin-B of the formula IA

[ia]

in which A and B are the same as those in formula VIII above, used as the starting material, the two 4 "and 6" hydroxyl groups thereof simultaneously with a divalent hydroxyl protecting group, such as that

65 isopropylidene group, protected, the 2 "hydroxyl group with a monovalent acyl group protecting the hydroxyl group, e.g. benzyl or acetyl, protected to a 2", 4 ", 6" tri-O-protected derivative of the formula IIB:

648564

22

[iib]

to form, in which A and B mean the same as above, the group

X

represents a divalent hydroxyl protecting group in which X and Y each represent a hydrogen atom, an alkyl group having 1 to 4 carbon atoms, an aryl group, e.g. is a phenyl group or an alkoxy group having 1 to 4 carbon atoms, or the group is dengruppe and D is a monovalent acyl group protecting the hydroxyl group. The 2 ", 4", 6 "-tri-0-protected derivative of the formula IIB obtained in this way is treated with aqueous acetic acid in order to select a cyclohexylidene group or a tetrahydropyranyl group.

split off from the 4 "and 6" hydroxyl groups, and the product thus partially deprotected is treated with a suitable sulfonylating reagent, e.g. p-toluene-30 sulfonyl chloride, reacted in pyridine to preferentially sulfonylate the 6 "hydroxyl group and a 6" -0-sulfonylated derivative of the formula IIIC

to form, in which A, B and D have the same meaning as above and G is a lower alkyl group, especially an alkyl group having 1 to 4 carbon atoms, an aryl group, e.g. a phenyl or p-methylphenyl group, or an aralkyl group, e.g. the benzyl group. The resulting 6 "-0-sulfonylated derivative is then treated with an alkali metal iodide or bromide to replace the 6" sulfonyloxy group (GSO3-) with the iodine or bromine group and thus the corresponding 6 "iodine or 6 "-bromo derivative (corresponding to a compound of formula IIIC, in which, however, the GSCb group is converted into the iodine or bromine atom), which is then reduced with hydrogen in the presence of a known hydrogenation catalyst, such as palladium, in order to reduce the halogen to effect, whereby the Penta-N-protected and 2 "-O-protected 3 ', 4', 6" -Tri-. deoxykanamycin B derivative of formula VIII, as desired, is obtained.

The preparation of the new compound of formula IV according to this invention is described below. The new compound of formula IV, e.g. IN - [(S) -4-amino-2-hydroxybutyryl] -5,3 ', 4'-trideoxy-6'-N-methylkanamycin-B, 55 1 -N- (3 - amino-2-hydroxypropionyl) - 5,3 ', 4' -trideoxy-6 '- N-methylkanamycin-B and IN - [(S) -5-amino-2-hydroxyva-leryl] -5,3', 4'-trideoxy-6'- N-methylkanamycin-B, can be prepared using the new compound 5,3 ', 4'-trideoxy-6'-N-methylkanamycin-B as the starting compound 60 obtained according to the present invention and condensing the 1st -Amino group of this starting compound with a corresponding a-hydroxy-co-amino fatty acid or a functional equivalent thereof.

65 Another object of the present invention is therefore a process for the preparation of a 1 -Na-hydroxy-co-aminoalkanolyl] -5,3 ', 4'-trideoxy-6'-N-methylkanamycin-B, as described above, of the formula IV

23

648564

co choh

I.

(ch2) nnh2

[iv]

in which n denotes 1, 2 or 3, which is characterized by (a) the 1-amino group of 5,3 ', 4'-trideoxy-6'-N-methyl-

is that you can kanamycin-B of the formula V

[v]

or a partially amino-protected derivative of 5,3 ', 4'-trideoxy-6'-N-methylkanamycin-B of the formula V'

[v ']

in which A is a hydrogen atom and at least one B is a 60 A 'monovalent amino protecting group, but the other B are hydrogen or at least one pair of A and B together B'. form a divalent amino protecting group, but the other A and B represent hydrogen, wherein the amino protecting groups represented by A and B may be the same or different, by reaction with an a-hydroxy-co-amino fatty acid or an amino-protected derivative thereof of formula VII

~ N (CH2) "CHCOOH

I.

OH

(VII)

65 in which A 'is a hydrogen atom and B' is a hydrogen atom or a monovalent amino protecting group or A 'and B' together are a divalent amino protecting group and n is 1, 2 or 3, or a functional derivative of the

648 564 24

bond VII aeylated to give the 1-N-acylated product of the formula IV '

h2n

(ch2) nn

[iv]

or formula IV "

in which A, B, A ', B' and n have the same meaning as above, and

(b) the remaining amino protecting groups, if present, are cleaved from the 1-N-acylated product of the formula IV 'or IV "in a known manner in order to obtain the compound of the formula IV.

This process may include a further step in which the compound of formula IV obtained is converted to a pharmaceutically acceptable acid addition salt by reaction with a pharmaceutically acceptable inorganic or organic acid in a known manner if desired.

The method described above can be carried out in the same way as the first method according to the invention.

When carrying out this third process, it is possible to use 5,3 ', 4'-trideoxy-6'-N-methylkanamycin B of the formula V as the starting compound, the amino groups of which are not protected at all, in the form of the free base or in the form of an acid addition salt thereof with a suitable acid, e.g. Hydrochloric acid. However, it is preferred to use as the starting compound a partially amino-protected derivative of 5,3 ', 4'-trideoxy-6'-N-me-so thylkanamycin-B of the formula V', in which all or part of the three amino groups and Methylamino groups with the exception of the I-amino group were protected with known amino protective groups and which can be obtained by introducing a known amino protective group into the compound V, as described in detail in connection with the first method according to the invention.

In this third process, the step of 1-N-acylation of the starting compound IV or IV 'and the step of 60 deprotection of the 1-N-acylated product IV' or IV "as well as the step of purifying the deprotected 1 -N-acylation product carried out in the same manner as described in connection with the corresponding steps of the first process.

65

The new compound of formula V, namely 5,3 ', 4'-trideoxy-6'-N-methylkanamycin-B, which as the starting compound in this third method according to the invention

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used, can be prepared starting from a known compound, namely 5,3 ', 4'-trideoxykana-mycin-B (Japanese Journal of Antibiotics, Vol. 32, p. 178 [1979]) and carrying out the N-methylation as in US Patent No. 3 925 353 or the "Journal of Antibiotics", 25, 743 (1972).

Another object of the present invention is a process for the preparation of 5,3 ', 4'-trideoxy-6'-N-

6'-amino group of this starting material in the same way 5 methylkanamycin-B of the formula V high:

[v]

which is characterized in that bonyl group or an aralkyloxycarbonyl group in the 6'-amino group of 5,3 ', 4'-trideoxykanamycin-B of (a) an alkyloxycarbonyl group, a cycloalkyloxycar formula X

introduces to the compound of formula XI

nhcoro

II J 0

generate in which R.3 represents an alkyl group having 1 to 6 carbon atoms, a cycloalkyl group having 3 to 6 carbon atoms or an aralkyl group, in particular a phenyl- (Ci-C4) -alkyl group, in particular benzyl, and

(b) reducing the compound of formula XI obtained with a metal hydride in an anhydrous organic solvent to obtain the compound of formula V.

In a preferred embodiment of this fourth method according to the invention, in the first stage an alkyloxycarbonyl group, a cycloalkyloxycarbonyl group or an aralkyloxycarbonyl group is introduced into the 6'-amino group of the starting compound (X), one of the urethane groups usually known as an amino protecting group Type can be. The introduction of the alkyloxycarbonyl, cycloalkyloxycarbonyl or aralkyloxycarbonyl group so (-COR3)

ii

O

into the 6'-amino group of the starting compound 5,3 ', 4'-tri-55 deoxykanamycin-B (X) is carried out in the same manner as described above for the preparation of the amino-protected derivative of the starting material (VF), which in the first according to the invention Procedure is used. Selective protection of the 6'-amino group can be achieved 60 because the 6'-amino group is the most readily reactive amino group in the kanamycin B compound (X). The group

(-COR3),

O

which in the 6'-amino group of the starting compound (X)

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26

should be introduced, preferably the methoxycarbonyl group, ethoxycarbonyl group or benzyloxycarbonyl group. In this way the 6'-N-alkyloxycar bonylated, 6'-N-cycloalkyloxycarbonylated or 6'-N-araI-kyloxycarbonylated product of the formula XI is formed.

The 6'-N-alkoxycarbonyl group, 6'-N-cycloalkyloxycarbonyl group or 6'-N-aralkyloxycarbonyl group thus introduced is reductively converted to a methyl group to achieve the 6'-N-methylation. This can be done by reducing the compound of formula XI with a metal hydride, e.g. Lithium aluminum hydride or diborane, in an anhydrous organic solvent, e.g. Tetrahydrofuran. This reduction can be carried out at a temperature of 40 to 90 ° C for 10 hours or longer.

The present invention is further illustrated by the following examples.

The residue was taken up in 100 ml of water. The aqueous solution was adjusted to pH 10.5 by adding aqueous ammonia and stirred at room temperature for 20 hours in order to split off the trifluoroacetyl group. The reaction solution obtained was concentrated to a volume of about 20 ml, adjusted to pH 7.5 by adding aqueous ammonia, diluted with 50 ml of water and then through a column of 150 ml of “Amberlite CG-50” resin (NH4 form , Product of Rohm & Haas Co., io USA). The column was washed successively with 750 ml of water and with 500 ml of 0.5 N aqueous ammonia and then eluted with 0.8 N aqueous ammonia. The eluate fractions containing the desired product were combined and evaporated to dryness, where 1.76 g (72%) IN - [(S) -4-amino-2-hydroxybutyryl] -5,3 ', 4' - trideoxykanamycin B monocarbonate monohydrate were obtained as a colorless powder. Overall yield: 32%.

example 1

Synthesis of 1 -N - [(S) -4-amino-2-hydroxybutyryl] - 20

5,3 ', 4' -trideoxykanamycin-B

(a) 4.36 g (10 millimoles) of 5,3 ', 4'-trideoxykanamycin-B was dissolved in 100 ml of dry dimethyl sulfoxide,

whereupon 10.5 g (48 millimoles) of zinc acetate [Zn (CH3CCh) 2-2H2O] was added. The resulting mixture was stirred at room temperature for 20 hours to form a complex of 5,3 ', 4'-trideoxykanamycin-B with the zinc cation. After adding 12.0 g (39.5 millimoles) of p-methoxy-benzyl-S-4,6-dimethylpyrimid-2-ylthiocarbonate (product of Kokusan Kagaku KK, Japan), the resulting mixture was stirred at 50 ° C. for T / 2 hours stirred to achieve Np-methoxybenzyloxycarbonylation. The reaction solution obtained was then poured into 1000 ml of water and the pH was adjusted to 11 by adding aqueous ammonia to cause decomposition of the zinc complex 35, whereby a precipitate failed. The precipitate was filtered off, washed with 500 ml of water and dissolved in 60 ml of a solvent mixture of chloroform-methanol - 17% aqueous ammonia (volume ratio 50: 10: 1). The solution was subjected to chromatography 40 on a column of 500 g of silica gel ("Wako-gel C-200"), which was developed with the same solvent mixture, 4.1 g (44%) of a colorless powder of 3.2 ', 6'-Tri-N-paramethoxybenzyloxycarbonyl-5,3 ', 4'trideoxykanamycin-B were obtained. 45

3.7 g (4 millimoles) of this colorless powder was dissolved in 50 ml of dimethyl sulfoxide and 0.95 ml (8 millimoles) of ethyl tri-fluoroacetate was added to the solution. The resulting mixture was stirred at room temperature for 5 hours to achieve 3 "-N-trifluoroacetylation, thereby obtaining 3,2 ', 6'-tri-N-paramethoxybenzyloxycarbonyl-3" -N-tri-fluoroacetyl-5.3 ', 4'-trideoxykanamycin-B was obtained.

(b) To the reaction solution containing the N-protected kanamycin B derivative obtained in step (a) above, 0.6 ml (4.4 millimoles) of triethylamine was added, followed by a solution of 1.9 g (6 millimoles) of N-hydroxysuccinic ester of (S) -4-tert-butoxycarbonylamino-2-hydroxy-butyric acid in 20 ml of dioxane. The mixture was stirred at room temperature for 19 hours, after which the reaction solution was poured into 500 ml of water to precipitate. The precipitate was filtered off and washed with 100 ml of water, whereby 10.9 g of a colorless powder were obtained. The powder was dissolved in 20 ml of 90% trifluoroacetic acid and the solution was left at room temperature for 50 minutes to give the amino protecting groups. The solution was then evaporated to dryness, and the

55

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65

Example 2

Synthesis of l-N- (3-amino-2-hydroxypropionyl) -5,3 ', 4'-trideoxykanamycin-B

(a) 435.5 mg (1 millimole) of 5,3 ', 4'-trideoxykanamycin-B was dissolved in 10 ml of dry dimethyl sulfoxide, followed by 1.05 g (4.8 millimole) of zinc acetate [Zn (CH3CCh) 2' 2H2O ] were added to the solution. The resulting mixture was stirred at room temperature for 23 hours. A solution of 937.3 mg (3.9 millimoles) of tert-butyl-S-4,6-dimethylpyrimid-2-yl-thiocarbonate in 5 ml of dimethyl sulfoxide was then added and the resulting mixture was stirred at 50 ° C. for 24 hours to achieve the N-tert-butoxycarbonylation. 15 ml of water were added to the reaction solution obtained, the pH was adjusted to 11 with aqueous ammonia, then 6.4 g of sodium chloride were added and the mixture was extracted with ethyl acetate (2 × 15 ml). The ethyl acetate extracts were combined and evaporated to dryness and the residue was taken up in 6 ml of dimethyl sulfoxide. The solution was mixed with 0.125 ml (1 millimole) of ethyl trifluoroacetate and the mixture was stirred at room temperature for 4 hours to effect the 3 "-N-trifluoroacetylation.

(b) To the reaction solution obtained in the step (a), which is 3,2 ', 6' -Tri-N-tert-butoxycarbonyl-3 "-N-tri-fioracetyl-5,3 ', 4' -trideoxykanamycin -B, 0.1 ml (0.7 millimoles) of triethylamine and 384 mg (1.05 millimoles) of N-hydroxysuccinimide ester of paramethoxybenzyloxoxycarbonylisoserine were added, and the resulting mixture was stirred at room temperature for 20 hours, followed by reaction solution with 10 ml of water was added and extracted with ethyl acetate (2 x 10 ml), the combined extracts were concentrated to a small volume and then 20 ml of 3N hydrochloric acid -50% methanol were added. The mixture was stirred at room temperature for 2 hours to give the to effect simultaneous cleavage of the two tert-butoxycarbonyl and para-methoxybenzyloxycarbonyl groups as amino protective groups.

The reaction solution obtained was adjusted to pH 10 by adding aqueous ammonia and then stirred at room temperature for 21 hours in order to enable the trifluoroacetyl group to be split off. The reaction solution thus obtained was evaporated to a small volume and then diluted with water, and the aqueous solution obtained was passed through a column on 25 ml of "Diaion WK-10S" (NW form, product of Mitsubishi Kasei K.K., Japan). The column was then washed with 125 ml of water and then eluted with 0.4 N aqueous ammonia. The eluate fractions which contained the desired product were combined

and evaporated to dryness to give 257 mg of 1 -N- (3-amino-2-hydroxypropionyl) -5,3 ', 4' -trideoxykanamycin-B-monocarbonate as a colorless powder. Overall yield: 42%.

Example 3

Synthesis of l-N - [(S) -4-amino-2-hydroxybutyryl] -5,3 ', 4', 6 "-tetradeoxykanamycin-B

(a) 419.5 mg (0.75 millimoles) of 5,3 ', 4', 6 "tetradeoxycana mycine-B-dicarbonate monohydrate was dissolved in 10 ml of dry dimethyl sulfoxide, followed by 1.05 g (4th , 8 millimoles) of zinc acetate [Zn (CH3CC> 2) 2 • 2HO] were added to the solution and the mixture was stirred at room temperature for 18 hours, followed by a solution of 1.44 g (6.0 millimoles) of tert. Butyl-S-4,6-dimethyl-pyrimid-2-ylthiocarbonate in 5 ml of dimethyl sulfoxide was added to the mixture and the resulting mixture was stirred for 25 hours at 50 ° C. 15 ml of water was added, and aqueous ammonia was added to the reaction solution obtained Adjusted to pH 11 and extracted with ethyl acetate (2x15 ml). 6.4 g of sodium chloride was added to the aqueous layer and further extracted with 20 ml of ethyl acetate. All of the ethyl acetate extracts were combined and evaporated to dryness, and the residue was taken up in 6 ml of dimethyl sulfoxide The solution was mixed with 0.125 ml (1 millimole) of ethyl trifluoroacetate and the G Then it was stirred at room temperature for 4 hours.

(b) To the reaction solution obtained in step (a) above, which contains 3,2 ', 6'-tri-N-tert-butoxycarbonyl-3 "-N-tri-fluoroacetyl-5,3', 4 ', 6 "tetradeoxykanamycin-B, 0.1 ml (0.7 millimoles) of triethylamine and 399.4 mg (1.05 millimoles) of N-hydroxysuccinimide ester of (S) -4-p-methoxybenzyloxycarbonylamino-2-hydroxy -butyric acid added. The resulting mixture was stirred at room temperature for 18 hours, after which the reaction solution was mixed with 10 ml of water and extracted with ethyl acetate (2 x 10 ml). The combined extracts were concentrated to a small volume and then mixed with 20 ml of 3N hydrochloric acid - 50% methanol. The mixture was stirred at room temperature for 2 hours to remove both the tertiary butoxycarbonyl group and the para-methoxy-benzyloxycarbonyl group. The reaction solution obtained was adjusted to pH 10 by adding aqueous ammonia and then stirred at room temperature for 20 hours in order to enable the trifluoroacetyl group to be split off. The reaction solution thus obtained was concentrated to a small volume and diluted with water, whereupon the aqueous solution obtained was passed through a column of 25 ml "Diaion WK-10S" (NH4 form). The column was then washed with 125 ml of water and then eluted with 0.32 N aqueous ammonia. The eluate fractions containing the desired product were combined and evaporated to dryness to give 303 mg IN - [(S) -4-amino-2-hydroxybutyryl) -5,3 ', 4', 6 "-tetradeoxykanamycin- B-dicar bonat as a colorless powder, overall yield: 63%.

Example 4

Synthesis of 5,3 ', 4', 6 "tetradeoxykanamycin-B

(a) Preparation of l, 3,2 ', 6', 3 "-penta-N-ethoxycarbonyl-3 ', 4' -dideoxykanamycin-B

4.64 g (10.3 millimoles) of 3 ', 4'-dideoxykanamycin-B was dissolved in a mixture of 45 ml of water and 45 ml of methanol, followed by the addition of 8 g (95.2 millimoles) of sodium bicarbonate, followed by slowly add 7.2 ml (75.6 millimoles) of ethyl chloroformate with ice cooling. The resulting mixture was kept at room temperature for 18

648564

Stirred for hours to effect N-ethoxycarbonylation. The reaction solution obtained was treated with 500 ml of water to separate out a precipitate. The precipitate was filtered off and washed with water to give 6.49 g (78%) of a colorless powder of the title compound.

(b) Preparation of 1,3,2 ', 6', 3 "-penta-N-ethoxycarbonyl-4", 6 "-0-isopropylidene-3 ', 4' -dideoxykanamycin-B

4.83 g (5.95 millimoles) of the product obtained in step (a) above were dissolved in 50 ml of dimethylformamide and the solution obtained with 30 mg (0.16 millimoles) of p-toluenesulfonic acid and 1.1 ml ( 9.0 millimoles) of 2,2-dimethoxypropane. The resulting mixture was stirred at room temperature for 25 hours to effect the 4 ", 6" -0-isopropylidation, whereupon the reaction solution was mixed with 1 ml (7.2 millimoles) of triethylamine and then evaporated to dryness, 5 , 1 g (100%) of the title compound were obtained as a pale yellow powder.

(c) Preparation of l, 3,2 ', 6', 3 "-penta-N-ethoxycarbonyl-4", 6 "-O-isopropylidene-2" -O-benzoyl-3 ', 4' -dideoxykana-mycin -B

5.1 g (6.0 millimoles) of the product from step (b) above was dissolved in 75 ml of pyridine, then 1 ml (8.6 millimoles) of benzoyl chloride was added and the mixture was stirred at room temperature for 5 hours The reaction solution thus obtained was mixed with 10 ml of water, stirred at room temperature and then evaporated to dryness. The residue was taken up in 250 ml of chloroform and the solution successively with 100 ml of 0.2 N hydrochloric acid and The chloroform layer was separated, dried over anhydrous sodium sulfate and evaporated to dryness to give 5.7 g (99%) of the title compound as a pale yellow powder.

(d) Preparation of l, 3,2 ', 6', 3 "-penta-N-ethoxycarbonyl-2" -O-benzoyl-3 ', 4' -deoxykanamycin-B

5.3 g (5.5 millimoles) of the product from step (c) above were dissolved in 80 ml of a mixture of acetic acid-methanol-water (volume ratio 3: 1: 1) and the solution obtained stood at room temperature for 21 hours left and then evaporated to dryness, 4.9 g (97%) of the title compound being obtained as a pale yellow powder.

(e) Preparation of 1,3,2 ', 6'3 "-penta-N-ethoxycarbonyl-2" -0-benzoyl-6 "-0-toxyl-3', 4'-dideoxykanamycin-B

1.5 g (1.63 millimoles) of the product obtained in step (d) above were dissolved in 28 ml of pyridine, 374 mg (1.96 millimoles) of paratoluenesulfonyl chloride were added and the mixture was stirred at room temperature for 28 hours. to effect the 6 "-0-tosylation. The reaction solution obtained was evaporated to dryness, the powdery residue (2.3 g) was taken up in 12.5 ml of dichloromethane and the solution obtained was subjected to column chromatography on silica gel (" Wako-gel C-200 », 200 g) The column was then washed with 1000 ml dichloromethane-ethanol (60: 1) and then developed with dichloromethane-ethanol (50: 1), 1.0 g (60%) a colorless powder of the title compound were obtained.

(f) Preparation of l, 3,2 ', 6', 3 "-penta-N-ethoxycarbonyl-2" -0-benzoyl-6 ".- iodine-3 ', 4'-dideoxykanamycin-B

900 mg (0.84 millimoles) of that obtained in step (e) above27

5

10th

IS

20th

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35

40

45

50

55

60

65

648564

28

The product was dissolved in 18 ml of dimethylformamide, 12.6 g (84 millimoles) of sodium iodide were then added, and the mixture was stirred at 95 ° C. for 2 hours to achieve 6 "iodination. The reaction solution obtained was dissolved in 250 Poured ml of water and the precipitate which had separated out was filtered off and then dissolved in 100 ml of chloroform .The chloroform solution was washed with 100 ml of 20% aqueous sodium thiosulfate solution and with 100 ml saturated aqueous sodium chloride solution. The separated chloroform layer was dried over anhydrous sodium sulfate and evaporated to dryness , whereby 817 mg (95%) of the title compound were obtained as a colorless powder.

(g) Preparation of l, 3,2 ', 6', 3 "-penta-N-ethoxycarbonyl-2" -Ó-benzoyl-3 ', 4', 6 "-trideoxykanamycin-B

773 mg (0.75 millimoles) of the product obtained from step (f) above was dissolved in 20 ml of dioxane, after which a small amount of Raney nickel W-2 was added and the mixture under a hydrogen pressure of 3.6 kg / cm 2 was hydrogenated for 23.5 hours using a Parr device. The reaction solution obtained was filtered to remove the catalyst and then evaporated to dryness to obtain 655 mg (97%) of a colorless powder of the title compound.

(h) Preparation of l, 3,2 ', 6', 3 "-penta-N-ethoxycarbonyl-2", 4 "-di-O-benzoyl-3 ', 4', 6 '-trideoxykanamycin-B

622 mg (0.69 millimole) of the product obtained in the above step (g) was dissolved in 30 ml of pyridine, after which 0.1 ml (0.86 millimole) of benzoyl chloride was added to the solution, and the mixture at room temperature for 25 hours The reaction solution obtained was then evaporated to dryness and the residue taken up in 50 ml of chloroform. The solution was washed successively with 10% aqueous potassium bisulfate solution (2 x 30 ml), saturated aqueous solution Washed sodium bicarbonate solution (2x30 ml) and saturated aqueous sodium chloride solution (1 x 30 ml), then dried over anhydrous sodium sulfate and finally evaporated to dryness to give 663 mg (96%) of the title compound as a colorless powder.

(i) Preparation of l, 3,2 ', 6', 3 "-penta-N-ethoxycarbonyl-2", 4 "-di-0-benzoyl-5-chloro-3 ', 4', 6" -trideoxykanamycin -B

600 mg (0.6 millimole) of the product obtained in the above step (h) was dissolved in 12 ml of pyridine, whereupon 0.1 ml (1.24 millimole) of sulfuryl chloride was added while cooling with ice, and the mixture was then kept at room temperature for 18 hours was stirred to effect 5-chlorination. The reaction solution thus obtained was mixed with 100 ml of water and extracted with 60 ml of chloroform. The chloroform extract was washed successively with 60 ml of 10% aqueous potassium bisulfate solution, 60 ml of saturated aqueous sodium bicarbonate solution and 60 ml of saturated aqueous sodium chloride solution, then dried over anhydrous sodium sulfate and finally evaporated to dryness. The powdery residue obtained (572 mg) was taken up in 5 ml of dichloromethane and chromatographed on a column of silica gel (“Wako-gel C-200”, 50 g). The column was washed with 500 ml dichloromethane and developed with a mixture of dichloromethane-ethanol (100: 1) to give 356 mg (58%) of a colorless powder of the title compound.

(j) Preparation of I, 3,2 ', 6', 3 "-penta-N-ethoxycarbonyl-

2 ", 4" -di-0-benzoyl-5,3 ', 4', 6 "-tetradeoxykanamycin-B

308 mg (0.13 millimoles) of the product obtained in step (i) above was dissolved in 5 ml of dioxane, a small amount of Raney nickel W-2 was added and the mixture obtained under a hydrogen pressure of 3.6 kg / cm2 was hydrogenated for 23.5 hours using a Parr device to remove the chlorine. The reaction solution obtained was filtered to remove the catalyst and then evaporated to dryness to give 295 mg (100%) of a colorless powder of the title compound.

(k) Preparation of 5,3 ', 4', 6 "tetradeoxykanamycin-B

120 mg (0.12 millimoles) of the product obtained in step (j) above was dissolved in a mixture of 1 ml of dioxane and 1 ml of water, after which 400 mg (1.27 millimoles) of barium hydroxide [Ba (OH2) • 8H2O] were added and the mixture obtained was subjected to the deprotection by heating to 110 ° C. for 15 hours under reflux. Then, carbon dioxide gas was introduced into the reaction solution to precipitate the barium carbonate, which was filtered off. The filtrate was mixed with 30 ml of water and passed through a column of 20 ml of “Amberlite CG-50” (NH4 form, product from Rohm & Haas Co., USA). The column was washed with 100 ml of water and 75 ml of 0.1 N aqueous ammonia and then eluted with 0.3 N aqueous ammonia. The eluate fractions containing the desired product were combined and evaporated to dryness to give 19.5 mg (29%) of 5,3 ', 4', 6 "tetradeoxykanamycin B-dicarbonate monohydrate as a colorless powder .

Example 5

Synthesis of 5,3 ', 5'-trideoxy-6'-N-methylkanamycin-B

(a) 4.0g (9.18 millimoles) of 5,3 ', 4'-trideoxykanamycin was dissolved in a mixture of 40 ml of water and 40 ml of methanol, followed by a solution of 5.8 ml (9.18 millimoles) of triethylamine and 2.72 g (11.02 millimoles) of 2- (tert-butoxycarbonyloxy-imino) -2-phenylacetonitrile (commercially available under the name "BOC-ON" from Aldrich Co., USA) in 40 ml of methanol was added . The resulting mixture was stirred at room temperature for 3 hours, whereupon the reaction solution containing the 6'-N-tert-butoxycarbonyl-5,3 ', 4'-trideoxykanamycin-B formed with 2 ml of a 17% solution aqueous ammonia solution mixed and evaporated to dryness. The residue was taken up in 100 ml of water and the solution was adjusted to pH 7.4 with 6N hydrochloric acid, whereupon it was washed with 50 ml of ethyl ether and then, via a column (20 mm internal diameter), from 100 ml of “Amberlite CG-50” ( NH4 + form, product from Rohm & Haas Co., USA). The column was washed with 300 ml of water and 150 ml of 0.1 M aqueous ammonia and then eluted with 0.2 M aqueous ammonia. The eluate was collected in 5 ml fractions and fractions no. 11 to 24 were combined and evaporated to dryness to give 2.1 g (42.5%) of 6'-N-tert-butoxy-carbonyl-5,3 ', 4'-trideoxykanamycin-B. Evaporation of the combined fractions No. 26 to 33 to dryness gave 1.42 g (35.6%) of unreacted 5,3 ', 4'-trideoxykanamycin-B.

(b) 1.85 g (3.45 millimoles) of the 6'-N-tert-butoxycarbonyl-5,3 ', 4'-trideoxykan-mycin-B obtained in step (a) above were dried in 60 ml Dissolved tetrahydrofuran and then decomposed with 1.31 g (34.5 millimoles) of lithium aluminum hydride and heated to 80 ° C. under reflux for 20 hours. The reaction mixture obtained was s

10th

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25th

30th

35

40

45

50

55

60

65

29

648 564

added dropwise to 200 ml of water, a precipitate precipitating out, which was filtered off. The filtrate was evaporated to remove the tetrahydrofuran and then adjusted to pH 6.5 by adding hydrochloric acid. The solution thus obtained was passed over a column (12 mm inner diameter) of 30 ml "Amberlite CG-50" (NH-i +) and the column was then washed with 150 ml of water and successively with 0.2 M aqueous ammonia (150 ml ), 0.3 M aqueous ammonia (150 ml) and 0.4 M aqueous ammonia. The eluate was collected in 6 ml fractions and the fractions no. 11 to 17 were combined and evaporated to dryness, 740 mg (39.9%) of unreacted 6'-N-tert-butoxycarbonyl-5,3 ', 4'-trideoxykanamycin-B being obtained. Fractions No. 37-55 were evaporated to dryness in a similar manner to give 592 mg (32.4%) of the title compound 5,3 ', 4'-trideoxy-6'-N-methylkanamycin-B.

Example 6

Synthesis of l-N - [(S) -4-amino-2-hydroxybutyryl] -5,3 ', 4'-trideoxy-6' -N-methylkanamycin-B

583 mg (1.10 millimoles) of 5,3 ', 4'-trideoxy-6'-N-methylkanamycin-B, prepared as described in Example 5, were dissolved in 10 ml of 90% aqueous dimethyl sulfoxide, followed by 1.16 g (5.28 millimoles) of zinc acetate [Zn (CH3CCh) 2 '2H2O] were added. The mixture was stirred at room temperature for 20 hours, after which a solution of 1.06 g (4.29 millimoles) of 2- (tert-butoxycarbonyloxyimino) -2-phenylacetonitrile in 5 ml of dimethyl sulfoxide was added and the mixture at 50 ° C was stirred for a further 6 hours to achieve the N-tert-butoxycarbonylation. The reaction solution obtained was mixed with 2 ml of 17% aqueous ammonia and then washed with 100 ml of water and with 50 ml of ethyl ether. The separated aqueous layer was adjusted to pH 11 with 17% aqueous ammonia and mixed with 2 g of sodium chloride, whereupon the resulting solution was extracted with ethyl acetate (3 x 100 ml). The combined ethyl acetate extracts were dried over anhydrous sodium sulfate and evaporated to dryness. The residue was chromatographed on a column (20 mm in diameter) of 50 g of silica gel (“Wako-gel C-200”, manufactured by Wako Yunyaku KK, Japan), using a mixture of chloroform - methanol - concentrated aqueous ammonia as the eluent (Volume ratio 30: 10: 1) was used. The eluate was collected in 10 ml fractions and fractions no. 25 to 50 were combined and evaporated to dryness, giving 607 mg (73.6%) 3,2 ', 6', tri-N-tert-butoxycarbonyl-5,3 ', 4'-trideoxy-6'-N -methylkanamycin-B were obtained.

590 mg (0.787 millimoles) of the product so obtained was dissolved in 10 ml of dimethyl sulfoxide, and 0.14 ml (1.18 millimoles) of ethyl trifluoroacetate was added to the obtained solution. The mixture was stirred at room temperature for 1.5 hours to obtain 3 "-N-trifluoroacetylene. The reaction solution obtained, which was 3,2 ', 6' -T ri-N-tert-butoxy carbonyl-3 "-N-trifluoroacetyl-5,3 ', 4'-trideoxy-6'-N-methylkanamycin-B was mixed with 0.16 ml (1.18 millimoles) of triethylamine and 420 mg (1.18 millimoles) of N- Hydroxysuccinimide esters of (S) -4-p-methoxybenzyloxycarbonylamino-2-hydroxybutyric acid dissolved in 4 ml of tetrahydrofuran were mixed and the mixture was stirred at room temperature for 4 hours to effect 1-N-acylation. 100 ml of water were then added to the reaction solution, which was then extracted with methyl acetate (2 × 100 ml). The combined extracts were dried over anhydrous sodium sulfate and evaporated to dryness, whereupon the 1-N-acy-

gated product in the form of the amino-protected derivative was obtained.

The 1-N-acylated product thus obtained was dissolved in 10 ml of 90% aqueous trifluoroacetic acid solution, and the solution was stirred at room temperature for 5 hours to remove the tertiary butoxycarbonyl group and the p-methoxy-benzyloxycarbonyl group, whereupon the reaction solution was evaporated to dryness . The residue was taken up in 30 ml of water and the solution was adjusted to pH 10 with 17% aqueous ammonia, after which it was stirred at room temperature for 18 hours to remove the trifluoroacetyl group. The reaction solution thus obtained was adjusted to pH 7.5 by adding 6N hydrochloric acid and passed through a column (13 mm inner diameter) of 20 ml of “Amberlite CG-50” (NH4 + form). The column was washed with 100 ml of water and eluted successively with 160 ml of 0.5 M aqueous ammonia and 100 ml of 0.8 M aqueous ammonia. The eluate was collected in 4 ml fractions and the 20 fractions No. 27 to 62 were combined and evaporated to dryness, giving 346 mg (71.8%) of the title compound IN - [(S) -4-amino-2-hydroxybutyryl] -5,3 ', 4'-trideoxy-6'- N-methylkanamycin-B was obtained in the form of its monocarbonate.

25th

Example 7

Synthesis of l-N- [3-amino-2-hydroxypropionyl] -5,3 ', 4'-trideoxy-6' -N-methylkanamycin-B

102 mg (0.136 millimoles) of 3,2 ', 6'-tri-N-tert-butoxycar-3o bonyl-5,3', 4'-trideoxy-6'-N-methylkanamycin-B, prepared as in Example 6 described, were dissolved in 3 ml of dimethyl sulfoxide and the solution obtained was mixed with 0.02 ml (0.204 millimoles) of ethyl trifluoroacetate. The mixture was stirred at room temperature for 1 hour. The resulting 35 reaction solution, which is 3,2 ', 6'-tri-N-tert-butoxycarbonyl-3 "-N-trifluoroacetyl-5,3', 4 '-trideoxy-6' -N-methylkana-mycin-B was mixed with 0.03 ml (0.204 millimole) of triethylamine and 75 mg (0.204 millimole) of N-hydroxysuccinimide ester of 3-p-methoxybenzyloxycarbonylamino-2-hydroxypro-4o pionic acid dissolved in 0.5 ml of tetrahydrofuran, and the mixture was mixed at room temperature Stirred for 7 hours to effect 1-N-acylation, then 10 ml of water was added to the reaction solution, which was then extracted with ethyl acetate (2x10 ml), the combined extracts were dried over anhydrous sodium sulfate and then dried evaporated, whereby 136 mg of the 1-N-acylated product were obtained in the form of the amino-protected derivative.

This 1-N-acylated product was dissolved in 3 ml of 90% aqueous 50% trifluoroacetic acid, and the solution was stirred at room temperature for 2 hours to remove the tertiary butoxycarbonyl group and the p-methoxybenzyloxycarbonyl group, and the reaction solution was dried was evaporated. The residue was taken up in 55 10 ml of water and the solution was adjusted to pH 10 with 17% strength aqueous ammonia and then stirred at room temperature for 16 hours in order to remove the trifluoroacetyl group. The reaction solution obtained in this way was adjusted to pH 6.4 by adding 6N hydrochloric acid and passed through a column (11 mm internal diameter) of 10 ml “Amberlite CG-50” (NH4 + form). The column was then washed with 30 ml of water and 30 ml of 0.2 M aqueous ammonia and then eluted with 0.5 M aqueous ammonia. The 65 eluate was collected in 1 ml fractions and fractions no. 4 to 7 were combined and evaporated to dryness, with 38.1 mg (45.4%) of the desired IN- [3-amino-2-hydroxypropionyl] -5,3 ', 4'-trideoxy-6'-N- methyl-

648564

30th

kanamycin-B was obtained in the form of its monocarbonate.

Example 8

Synthesis of l-N - [(S) -5-amino-2-hydroxyvaleryl] -5,3 ', 4'-trideoxy-6' -N-methylkanamycin-B

The same procedure as described in Example 7 was repeated, but using 76 mg (0.204 millimoles) of N-hydroxysuccinimide ester of (S) -5-p-methoxyben-

zyloxycarbonylamino-2-hydroxyvaleric acid instead of the 3-p-methoxybenzyloxycarbonylamino-2-hydroxy-propionic acid N-hydroxysuccinimide ester for the reaction with the 3,2 ', 6'-tri-N-tert-butoxycarbonyl-3 "-N -trifluoroacetyl-s 5,3 ', 4'-trideoxy-6'-N-methylkanamycin-B.

After eluting the “Amberlite CG-50” column, the eluate fractions No. 11 to 18 combined and evaporated to dryness to give 47.2 mg (53.8%) of the title compound as monocarbonate.

B

Claims (13)

648564 1. Compounds of formula I. PATENT CLAIMS [I] in which R2 represents a hydrogen atom or a methyl group, R is a hydroxyl group or a hydrogen atom, where R2 is not a methyl group if R is hydrogen Ri represents a hydrogen atom or an a-hydroxy-co-amino-alka, as well as their pharmaceutically acceptable acid addition salts. noyl group of the formula -CO-CH- (CH2) n-NH2 I. OH in which n is 1, 2 or 3, and 6 " 2. Compound according to claim 1, namely a 1 -N- [a-hydroxy-o-aminoalkanoyl] -5,3 ', 4' -trideoxykana-mycin-B or lN- [a-hydroxy-co-aminoalkanoyl] -5 , 3 ', 4', 6 "-30 tetradeoxykanamycin-B of formula II choh (ch2) nnh2 [ii] in which R is a hydroxyl group or a hydrogen atom 3. Compound according to claim 1, namely one and n is 1, 2 or 3, and their pharmaceutically IN- [a-hydroxy-a) aminoalkanoyl] -5,3 ', 4 "- trideoxy-6'-N- acceptable acid addition salts. methylkanamycin-B of formula IV 648564 choh i (ch2) nnh2 nhch: [iv] in which n is 1, 2 or 3, and their pharmaceutically acceptable acid addition salts. 4. A compound according to claim 2, namely 1-N - [(S) -4-amino-2-hydroxybutyryl] -5,3 ', 4' -trideoxy-kanamycin-B, 1 -N- [3-amino-2 -Hydroxypropionyl] -5,3 ', 4' -trideoxykana-mycin-B or IN - [(S) -4-amino-2-hydroxy-butyryl] -5,3 ', 4', 6 "-tetradeoxykanamycin-B . 5. A compound according to claim 3, namely 1-N- [3-25 amino-2-hydroxypropionyl] -5,3 ', 4'-trideoxy-6'-N-methyl- kanamycin-B, 1 -N - [(S) -4-amino-2-hydroxybutyryl] -5,3 ', 4'-trideoxy-6'-N-methylkanamycin B or lN - [(S) -5-amino -2-hydroxy valeryl] -5,3 ', 4'-trideoxy-6' -N-methyl-kanamycin-B. 6. A compound according to claim 1, namely 5,3 ', 4', 6 "-30 tetradeoxykanamycin-B of the formula III 5 'nh: [iii] as well as their pharmaceutically acceptable acid addition salts. 7. A compound according to claim 1, namely 5,3 ', 4'-trideoxy-6'-N-methylkanamycin-B of the formula V. h2n and their pharmaceutically acceptable acid addition salts. 8. Process for the preparation of an l-N- [a-hydroxy-co- aminoalkanoyl] -5,3 ', 4'-trideoxykanamycin-B or 1-N- [a-hydroxy- (o-aminoalkanoyl] -5,3', 4 ', 6 "-tetradeoxykan-mycin-B of formula II 648564 4th co i - choh i (ch2) nnh2 [ii] in which R is a hydroxyl group or a hydrogen atom (a) the 1-amino group of 5,3 ', 4' -trideoxykanamycin-B and n is 1, 2 or 3, characterized in that one or 5,3 ', 4', 6 "tetradeoxykanamycin-B of formula VI in which R has the same meaning as above, or a partially amino-protected derivative of 5,3 ', 4'-trideoxykana- [vi] mycin-B or5,3 ', 4', 6 "tetradeoxykanamycin-B of formula VI ' b [vi '] in which R is a hydroxyl group or a hydrogen atom 60 non-protected derivative thereof of the formula VII and A is a hydrogen atom and at least one B is a monovalent amino protective group, but the other B are each a hydrogen atom, or at least one pair of A and B together are a divalent amino protective group, but the other A and B each represent a hydrogen atom 65, where amino protecting groups represented by A and B can be the same or different, by reaction with an a-hydroxy-co-amino fatty acid or an amino B '. ; N (CH2) "CHCOOH OH (VII) in which A 'is a hydrogen atom and B' is a hydrogen atom or a monovalent amino protecting group or A 'and 5 648564 B 'together a divalent amino protecting group and n 1, compound of formula VII acylated to mean 1-N-acylated 2 or 3, or with a functional derivative of the product of formula II' 6 " or formula II " (ch2) nn in which R, A, B, A ', B' and n remove the same meaning II "to obtain the compound of formula II. tion as above, and (b) the remaining amino protecting groups, if any, from the 1-N-acylated product of the formula II 'or the formula IV h0- 9. Process for the preparation of a 1-N- [a-hydroxy-co-aminoalkanoyl] -5,3 ', 4' -trideoxy-6 '-N-methylkanamycin-B no choh i (ch2) nnh2 [iv] 648564 in which n means 1, 2 or 3, characterized in that one (a) the 1-amino group of 5,3 ', 4'-trideoxy-6'-N-methyl-kanamycin-B of formula V. [v] or a partially amino-protected derivative of 5,3 ', 4' -trideoxy-6 '-N-methylkanamycin-B of the formula V' / ch3 b 1 [v '] in which A is a hydrogen atom and at least one B is an atom, wherein the amino protecting groups which are monovalent amino protecting groups from A but the other B and B are the same or different, each represents a hydrogen atom, or at least a pair of reaction with an a-hydroxy-co-amino fatty acid or A and B together represent a divalent amino protective group 35 an amino-protected derivative thereof of the formula VII, but the other A and B each represent a hydrogen K B'- : N (CH2) n CHCOOH I. OH (vii) in which A 'represents a hydrogen atom and B' represents a hydrogen 2 or 3, or acylated with a functional derivative of the atom or a monovalent amino protecting group or A 'and compound of the formula VII to give the 1-N-acylated B' together a divalent Amino protecting group and n 1, 45 product of formula IV ' hn co I choh (ch2) nn a 'B1 [IV *] 648564 or formula IV " (ch2) nn ■ b ' [iv "] in which A, B, A ', B' and n remove the same meaning as above or IV "to have the compound of formula IV, and 25th (b) the remaining amino protecting groups, if any, from the 1-N-acylated product of the formula IV ' receive. 10. Process for the preparation of 5,3 ', 4', 6 "tetradeoxy-kanamycin-B of the formula III nh ' [iii] characterized in that 2 "-0-protected derivatives of 3 ', 4', 6" -trideoxykana- (a) the 4 "hydroxyl group of a penta-N protected and mycin-B of formula VIII ijit CH ^ [viii] in which A is a hydrogen atom and B is a monovalent amine-65 acyl group of the same type as the hydroxyl-protecting no-protecting group or A and B together are a divalent group D which is in the 2 "position of the compound VIII before-amino-protecting group and D is a hydroxyl-protecting acyl group, protects to the 4 "-O-protected compound of the group, with a monovalent hydroxyl protective formula VIII ' 648564 in which A, B and D have the same meaning as above, (b) reacting the 4 "-0-protected compound of formula VIII 'with sulfuryl chloride to convert the 5-hydroxyl group through a [VIII *] 15 to replace chlorine atom to obtain the corresponding 5-chloro derivative, (c) reducing the chlorine derivative to remove the 5-chloro group and a protected derivative of 5,3 ', 4', 6 "tetra-deoxykanamycin-B of Formula IX b s in which A, B and D have the same meaning as above, and (d) the remaining hydroxyl protecting groups and the remaining amino protecting groups from the compound of [ix] Formula IX removed to obtain the compound of Formula III 35. 11. Process for the preparation of 5,3 ', 4'-trideoxy-6'-N-methylkanamycin-B of the formula V h oh h [v] characterized in that one bonyl group or an aralkyloxycarbonyl group in the 55 6'-amino group of 5,3 ', 4'-trideoxykanamycin-B of (a) an alkyloxycarbonyl group, a cycloalkyloxycar formula X NH2 [X] 9 introduces to obtain the compound of formula XI 648564 [xi] in which R.3 denotes an alkyl group with 1 to 6 carbon atoms, a cycloalkyl group with 3 to 6 carbon atoms or an aralkyl group, and (b) reducing the compound of formula XI with a metal hydride in an anhydrous organic solvent to obtain the compound of formula V. 12. Antibacterial preparation, characterized in that it is an 1-N- [a-hydroxy-co-amino- (C3-C5L) -alkanoyl] -5,3 ', 4' -trideoxykanamycin-B as an active ingredient 1 -N- [a-Hydroxy-co-amino- (C3-Cs) -alkanoyl-5,3 ', 4', 6 "-tetra-deoxykanamycin-B or a pharmaceutically acceptable acid addition salt thereof or 5,3 ', 4 ', 6 "-tetradeoxykana-mycin-B or a pharmaceutically acceptable acid addition salt thereof in an antibacterially effective amount to inhibit the growth of bacteria, together with a carrier. 13. Antibacterial preparation, characterized in that it is an lN- [a-hydroxy-co-amino- (C3-C5) -alkanoyl-5,3 ', 4' -trideoxy-6 '-N-methylkanamycin as an active ingredient. B or 5,3 ', 4' -trideoxy-6 '-N-methylkanamycin-B or a pharmaceutically acceptable acid addition salt thereof in an antibacterially effective amount to inhibit the growth of bacteria, together with a carrier.