CH618180A5 - Process for the preparation of a derivative of the antibiotic XK-62-2. - Google Patents

Description

The invention thus relates to a process for the preparation of a compound of the formula A.

io

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(A)

ch3 oh where Rß is the grouping

45

OH O

HjjN — CH2 — CH2 — CH — C—

is or a pharmaceutically acceptable acid addition salt 50 thereof, characterized in that in a compound of formula A, in which R ,,. Is hydrogen, the free amino groups acylated by reaction with a compound of the formula

Y1Y2N-CH., - CH, -CHOH-COZ 55

where Z

'"Û •

5

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Cl, Br, I or OH and wherein Yx is hydrogen and Y2 is a protecting group selected

R.

CH O t 3 n

çj; -ch2-o-c-, ch3-c-o-c ~

O

IT

ch -o-c-3 _

O

n

^ 3-0îr2 "C-

O

* n c2v ° - ° - -

CH.

Chip or s—

1Î02

is

where R-! and R2 are the same or different and are hydrogen, OH, NO2, Cl, Br, I, alkyl having 1-5 carbon atoms or alkoxy having 1-5 carbon atoms and R3 is hydrogen, Cl, Br or I, or Yj and Y2 are one Form phthaloyl group to form a mixture of compounds of formula (C)

C-Hj where

(1) R4 'Y1Y2N-CH, -CH., - CH0H-C0- and R =' and Ru 'are hydrogen,

(2) R (i 'Y1Y2N-CH2-CHOH-CO- and R,' and R5 'represent hydrogen,

(3) R.j 'and Rr; Y ^ .N-CH ^ CH.-CHOH-CO- and R ,; ' Are hydrogen or

(4) R, 'and R ,.' Y ^ N-CH.-CH.-CHOH-CO- and R, 'is hydrogen,

where Y and Y2 have the above meaning, the protective group Yj, removed, the compound A is isolated from the mixture, or first of all a mixture from the mixture in which R.sup.2 '' Y, Y2N-CH. and R4 'and Rr,' are hydrogen, isolated and then the protective group Y. removed, and optionally converting a compound A thus obtained with a pharmaceutically acceptable acid into an acid addition salt.

In the process according to the invention, a mixture of compounds is obtained which, in the case of the following numbers

(1) and (2) one, or in the case of numbers (3) and (4) two

25 acyl groups, i.e. a-Hydroxy-3-aminobutyryl groups instead of the three free amino groups which had the antibiotic XK-62-2, bound in the 1-position or in the 2'- and 1- or 2'- and 3- Position.

30 These derivatives of XK-62-2 thus include

(1) a compound I in which one of the hydrogen atoms of the amino group bonded to the 2'-carbon atom of XK-62-2 is substituted by

35 O OH

!! I.

- C - CH - CH, - CH2 - NH2;

(2) a compound II in which one of the hydrogen atoms

40 of the amino group bonded to the 1-carbon atom of XK-62-2 is substituted by

O OH

II I

45 - C-CH-CH2-CH2NH,;

(3) a compound III in which one of the hydrogen atoms of the amino group bonded to the 2'-carbon atom and a hydrogen atom of that amino

50 nogroup, which is bonded to the 1-position carbon atom of XK - 62-2, are substituted by

O OH

II I

55-C-CH-CH2-CH2-NH2;

and (4) a compound IV, wherein one of the hydrogen atoms of the amino group attached to the 2'-carbon atom and a hydrogen atom of the amino group

60 pe, which is bonded to the 3-position carbon atom of XK-62-2, are substituted by

O OH

II I

65-C-CH-CH2-CHm-NH2.

The individual figures are discussed in more detail below.

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Figure 1 illustrates the IR absorption spectrum of 2'-N- [L - (-) - a-hydroxy-Y-aminobutyrylJ-XK-62-2 (Compound I);

Figure 2 illustrates the NMR spectrum of 2'-N- [L - (-) - a-hydroxy-Y-aminobutyryl] -XK-62-2;

Figure 3 illustrates the IR absorption spectrum of l-N- [L - (-) - a-hydroxy-y-aminobutyrylJ-XK-62-2 (Compound II);

Figure 4 illustrates the NMR spectrum of 1-N- [L - (-) -a-hydroxy-Y-aminobutyryl] -XK-62-2;

Figure 5 illustrates the IR absorption spectrum of l-N-2'-N-bis- [L - (-) - a-hydroxy-Y-aminobutyryl] -XK-62-2 (Compound III);

Figure 6 illustrates the NMR spectrum of l-N-2 '- N-bis- [L - (-) - a-hydroxy-Y-aminobutyryl] -XK-62-2;

Figure 7 illustrates the IR absorption spectrum of 3-N-2'-N-bis- [L - (-) - a-hydroxy-Y-aminobutyryl] -XK-62-2 (Compound IV) and

Figure 8 illustrates the NMR spectrum of 3-N-2'-N - bis- [L - (-) - a-hydroxy-Y-aminobutyryl] -XK-62-2.

The acylating agents used can be derived from the α-hydroxy-Y-aminobutyric acid or from a functionally equivalent compound, to produce:

1) a compound IA, in which one of the hydrogen atoms of the amino group of XK-62-2 bonded to the 2'-carbon atom is substituted by

O OH

II 1 yt

—C— CH — CH2 — CH., - Nd;

y2

(2) A compound IIA in which one of the hydrogen atoms of the amino group bonded to the 1-carbon atom of XK-62-2 is substituted by

O OH

II I Y1

—C — ch — ch., - ch. — nc; ;

Y2

(3) a compound IIIA, wherein one of the hydrogen atoms of the amino group bonded to the 2'-carbon atom and one of the hydrogen atoms on the am

1-position carbon atom-chained amino group of XK-62-2 are substituted by

O OH

IM Y,

—C — ch — ch., - ch., - n c; ;

Y2

and (4) a compound IVA, wherein one of the hydrogen atoms of the amino group bonded to the 2'-carbon atom and a hydrogen of the amino group bonded to the 3-carbon atom of XK-62-2 is substituted by o oh

II I Yt

—C — ch — ch, —ch., - nc!

Y2

where Y, and Y2 have the above meaning.

It can be seen that acylating agents with other groups can also be selected for the preparation of compounds IA, IIA, IIIA and IVA. It is also possible to choose other derivatives of a-hydroxy-Y-aminobutyric acid which are functionally equivalent in terms of introducing an a-hydroxy-Y-ami-nobutyryl group to a free amino group.

The acylation methods with functionally equivalent acylating agents are described in M. Bodansky and co-workers, Synthesis, p. 435 (1972) and M. Bodansky and co-workers, Peptide Synthesis, pp. 75-135 (1966) (John Wiley & Sons, Inc., USA).

The acylation is advantageously carried out using an acylating agent of the formula

0

OH O

: N-CH2-CH2-CH-C-O-N

wherein Yx and Y2 have the meanings given above. In this case, a compound of formula

O

Yx II

^ N-CH2-CH2-CH-C-OH Y2 I

OH

be reacted with N-hydroxysuccinmide in the presence of dicyclohexyl-carbodiimide, a compound of the formula

0

OH O

: N-CH2-CH2-CH-C-O-N

is created and the resulting connection can then be isolated and implemented with XK-62-2.

According to another alternative, a compound of the formula

OH O

Yi | II

^: N — ch2 — ch2 — ch — c — oh

Y,

wherein yi and y2 have the above meanings, reacted with N-hydroxysuccinimide and dicyclohexylcarbodiimide and the resulting reaction mixture is reacted with XK-62-2.

According to another embodiment, the acylation can be carried out by adding dicyclohexylcarbodiimide to a mixture of XK-62-2 and a compound of the formula

OH O

y, | II

^ n — ch. — ch. — ch — c — oh

Y2

In general, 0.4-2.5 moles, preferably 0.7-1.5 moles, of the acylating agent are used per 1 mole of XK-62-2. The reaction is conveniently carried out at -50 to 50 ° C, preferably -20 to 20 ° C for 15 minutes to 24 hours, advantageously 5-15 hours.

The following solvents can be selected for the reaction, for example, tetrahydrofuran, dimethylacet

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amide, dimethylformamide, lower alcohols, dioxane, ethylene glycol dimethyl ether, pyridine and water. One or a mixture of two or more of these solvents can be used.

A mixture of ethyl alcohol and water in a ratio of 2: 1 is preferably used.

The intermediate compounds IA, IIA, IIIA and IVA prepared in the above manner can be isolated or purified for use in the second reaction. However, it is preferred that the reaction mixture be used as such without isolation and purification of the acylated compounds. Following the subsequent reaction, the compounds I, II, III and IV prepared from the mixture of intermediate compounds are isolated and purified. The latter method is advantageous because it simplifies the process and increases the yield.

If necessary, the compounds IA, IIA, IIIA and IVA can be conveniently isolated and purified by known methods, for example by column chromatography using an adsorbent such as an ion exchange resin, silica gel, alumina, cellulose, "Sephadex" etc., and with the help thin layer chromatography using silica gel, alumina, cellulose, etc., as is well known.

The compounds IA, IIA, IIIA and IVA are either isolated or treated in a mixture in order to split off the protective groups Y-t and Y2 to give the compounds I, II, III and IV.

The protecting groups can be split off in a known manner. If the protective group is a phthaloyl group, it can be split off with hydrazine; if the protecting group is a carbomethoxy group or carboethoxy group, the elimination can be carried out with barium hydroxide; if the protective group is a tertiary butoxycarbonyl group, it can be split off with formic acid or trifluoroacetic acid; if the protective group is an orthonitrophenyl-sulfenyl group, the cleavage can be carried out with acetic acid or hydrochloric acid and the protective group is a chloroacetyl group, the cleavage can be carried out with 3-nitro-pyridin-2-thione, according to K-Und-heim and Collaborator: Journal of the Chemical Society, Parkin Transactions, Part I, p. 829 (1973).

In a preferred embodiment, the protective group is one of the formula

- 'C1I

R

wherein R [and R2 have the meaning given above, and the cleavage takes place by hydrogenolysis in the presence of a metal catalyst such as palladium, platinum, rhodium or Raney nickel, preferably a palladium catalyst on a support of activated carbon in at least one solvent selected from the group consisting of tetrahydro-furan , Dimethylacetamide, dimethylformamide, lower alcohols, dioxane, ethylene glycol dimethyl ether, pyridine and water, preferably in a mixture of water and methanol 1: 1; in the presence of a small amount of hydrochloric acid, hydrobromic acid, hydroiodic acid and acetic acid, preferably acetic acid, at room temperature and atmospheric pressure.

A technique used in peptide synthesis is commonly used to introduce an easily removable protecting group. The protection of amino groups with such protective groups and their cleavage are by M.

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Bodansky and co-workers: Peptide Synthesis, pp. 21-41 (1966) (John Wiley & Sons, Inc., USA) and A. Kapoor: Journal of pharmaceutical Sciences, Vol. 59, pp. 1-27 (1970) .

The compounds I, II, III and IV thus prepared, which either from isolated and purified intermediate compounds IA, IIA, IIIA and IVA or from a mixture of compounds IA, IIA, IIIA and. IVA were prepared, isolated from the reaction mixture and purified. For example, the compounds can be isolated and purified by column chromatography using an adsorbent such as ion exchange resins, silica gel, alumina, cellulose, etc., and thin layer chromatography using silica gel, alumina, cellulose, etc.

In the present process, compounds I, II,

III and IV obtained in their 1, d or dl form, but the compounds are preferably obtained in their 1 form.

The compounds I, II, III and IV can be converted into pharmaceutically acceptable, non-toxic acid addition salts, namely mono-, di-, tri-, tetra- or pentasalts, by reaction with suitable acids. Non-toxic acids include inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, carbonic acid, etc., and organic acids such as acetic acid, fumaric acid, malic acid, citric acid, mandelic acid, tartaric acid, ascorbic acid, etc.

The derivatives I, II, III and IV of XK-62-2 have an excellent antibacterial activity. It is particularly noteworthy that the derivatives have a strong antibacterial activity against strains of Escherichia coli which has such an R factor which indicates resistance to known amino-glycoside antibiotics.

The following Table I illustrates the antibacterial spectrum of kanamycin A, gentamicin Cla, XK-62-2 and compounds 2'-N- [L - (-) - a-hydroxy-y-aminobutyryl] -XK - 62-2 (Compound I), IN- [L - (-) - a-Hydroxy-Y-aminobutyryl] -XK-62-2 (Compound II), IN-2'-N-bis- [L - (-) -a-hydroxy-y-aminobutyryl] -XK-62-2 (compound III) and 3-N - 2'-N-bis- [L - (-) - a-hydroxy-y-aminobutyryl] - XK-62-2 (Compound IV) against various gram-positive and gram-negative bacteria as determined by the agar dilution method at pH 8.0.

A comparison of the minimum inhibitory concentration in Table 1 shows that compounds I, II, III and

IV show strong antibacterial activity. It is particularly noteworthy that these compounds have a strong antibacterial activity, in particular against Escherichia Coli KY 8327 and 8348.

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TABLE 1

Antibacterial spectrum (minimum inhibitory concentration, mcg / ml)

Tribes

Kanamycin gentamicin

A C, "XK-62-2

I.

Compounds II III

IV

Pseudomonas aeruginosa BMH 1

Staphylococcus aureus ATCC 6538P

Bacillus subtüis No. 10707

Proteus vulgaris ATCC 6897

Shigella sonnei ATCC 9290

Salmonella typhosa ATCC 9992

Krebsiella pneumoniae ATCC 10031

Escherichia coli ATCC 26

Escherichia coli KY 8327

Escherichia coli KY 8348

0.13 0.52 25 2.08 8.34 8.34

0.021 0.004 0.088 0.05 0.016 0.065 0.130

0.021 0.004 0.004 0.05 0.004 0.008 0.016

0.16 0.033 0.033 0.39 0.065 0.521 0.521

0.16 0.033 0.033 0.39 0.033 0.260 0.260

0.088 0.016 0.008 0.20 0.008 0.065 0.130

0.042 0.016 0.004 0.10 0.008 0.065 0.065

0.16 0.033 0.016 0.39 0.004 0.065 0.130

1.04 2.08 1.04 4.10 0.016 0.065 0.260

0.041 1.04 1.04 0.05 0.004 0.065 0.130

In the above table, Escherichia coli KY 8327 and KY 8348 intracellularly produce gentamicin adenylyl transferase and gentamycin acetyl transferase of type I. The former bacterium inactivates kanamycin and gentamicin by adenylation and the latter inactivates gentamicin 40 by acetylation.

The antibacterial spectrum of IN - [L - (-) - a-Hydroxy - y-aminobutyry 1] -XK-62-2 (Compound II) compared to Kanamycin A, Gentamicin complex (C ^ Cla and C2) and XK-62-2, determined by the agar dilution method at 45 pH 7.2, is shown in Table 2 below.

TABLE 2

Antibacterial spectrum of l-N- [L - (-) - a.-Hydroxy - ^ - aminobutyryl] -XK-62-2 lowest inhibitory concentration mcglml

Tribes

Kanamycin A

Gentamicin complex (Cl CIa and C2)

XK-62-2

Compound II (L type)

Staphylococcus aureus 209 P.

0.2

<0.05

0.1

0.1

Staphylococcus aureus Smith

0.2

<0.05

<0.05

0.1

Bacillus subtilis ATCC 6633

0.2

<0.05

<0.05

0.1

Sorcina lutea ATCC 9341

6.25

0.2

0.4

0.4

Escherichia coli T-2

1.56

0.4

0.4

0.4

Escherichia coli T-5

1.56

0.4

0.4

0.4

Escherichia coli Ky 83271

50

12.5

12.5

0.2

Escherichia coli Ky 83492

100

6.25

3.12

0.2

9

TABLE 2 (continued)

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Tribes

Kanamycin A

Gentamicin complex

(Cl. Cla and C2>

XK-62-2

Compound II (L type)

Escherichia coli Ky 83483 Escherichia coli Ky 83494 Pseudomonas aeruginosa BmH No. 1 Pseudomonas aeruginosa Ky 85103 Pseudomonas aeruginosa Ky 85116 Pseudomonas aeruginosa Ky 8512T Pseudomonas aeruginosa Ky 8516s Pseudomonas providencia sp. 1649 Klebsiella pneumoniae No. 8045 Proteus Mirabilis 1287 Proteus vulgaris 6897 Proteus rettgeri KY 4288 Proteus morganii KY 4298

0.78> 100 12.5 100 100 12.5> 100> 100 0.4 6.25 3.12 0.78 1.56

3.12 0.2 0.4 3.12 50 0.4 3.12 50 0.2 1.56 0.78 0.78 6.78

12.5 0.4 0.78 1.56 100 0.78 3.12 100 0.1 0.78 0.78 0.4 0.4

0.1 0.2 1.56 3.12 3.12 0.78 3.12 12.5 0.2 3.12 3.12 0.78 0.78

1 produces gentamicin adenylyl transferase

2 produces gentamicin adenyltransferase and neomycin kanamycin phosphotransferase type II

3 produces type I gentamicin acetyltransferase

4 produces neomycin-kanamycin phosphotransferase type I 3 produces kanamycin acetyltransferase

0 produces gentamicin acetyltransferase type I and neomycin

Kanamycin phosphotransferase type I 7 produces neomycin kanamycin phosphotransferase type I and

Type II and streptomycin phosphotransferase • s likely to produce kanamycin acetyltransferase 9 produces gentamicin acetyltransferase type II

The minimum inhibitory concentration of Gentamicin 35 administered an effective dose of 1.6-6 mg / kg per day

Complex to Proteus morganii KY 4298 is 0.78. will.

It can be seen from Table 2 above that the acute toxicity LD30 of l-N- [L - (-) - a-hydroxy-y-

generated according to l-N- [L - (-) - a-hydroxy-Y-aminobuytryl] - -aminobutyryl] -XK-62-2 (compound II) in mice

-XK-62-2 a very strong antibacterial activity against- 250 mg / kg with intravenous injection, while that of

has various bacteria that are resistant to 40 XK-62-2 and the gentamicin complex, a mixture to one of the gentamicin antibiotics and XK-62-2, and by Cj, Cla and Co 93 mg / kg or Is 72 mg / kg,

the gentamicin adenyl transferase and / or gentamicin. The method according to the invention is described below with

Acetyltransferase of type I and type II intracellu- help of preferred embodiments explained in more detail, generate, and thereby the gentamicin antibiotics and that

Deactivate XK-62-2. 45 Example 1

Based on various protection tests, production of 2'-N- [L - (-) - a-hydroxy-Y-carbobenzoxy-

against infections induced by strains above-aminobutyryl! -XK-62-2 (compound IA), l-N- [L - (-) - a-Hy-

Resistant bacteria, it was found that the inventive hydroxy-Y-carbobenzoxyaminobutyryl] -XK-62-2 (compound according to the compound prepared the a-hydroxy-y-aminobuty IJA), IN-2'-N-bis- [L - (-) - a-Hydroxy-Y-carbobenzoxyamino-

ryl group in vivo and a significantly larger anti-50 butyryl] -XK-62-2 (compound IIIA) and 3-N-2'-N-bis- [L-

bacterial activity, as gentamicin antibiotics and - (-) - a-hydroxy-Y-carbobenzoxyaminobutyryl] -XK-62-2

XK-62-2 has. (Compound IVA).

From the tables above it can further be seen that the 2,778 g (6.0 mmol) of XK-62-2 in 30 ml of aqueous

Compounds I, II, III and IV dissolved a particularly strong antigen 50% tetrahydrofuran. To this solution bacterial activity against a variety of gram- 55 is then a solution of 2.94 g (84 mmol) of the N-hydroxy-

positive and gram-negative bacteria, including those succinimide esters of L - (-) - a-hydroxy-Y-carbobenzoxy-

which are resistant to aminoglycoside antibiotics, aminobutyric acid in 20 ml of tetrahydrofuran with stirring and gene. It is therefore expected that these compounds will be used while maintaining the temperature at -5 to 0 ° C.

treating various infections in humans and giving. The production of the first connection from L - (-) - a-

in animals that are caused by inflammation-causing bacteria - 60 Hydroxy-Y-aminobutyric acid is published in the Journal of Antibiotics,

are called, are effective. Vol. XXV. Pp. 695-708 (1972) and the production of L - (-) -

For example, it is believed that compounds for -a-hydroxy-Y-aminobutyric acid in Tetrahedron Letters, S. describe the treatment of urinary tract and AT 2625-2628 (1971) infections. The addition is complete in one hour, induced by Staphylococcus aureus, Escheri and the mixture is left to react overnight, chia coli, Pseudomonas aeruginosa and strains of genus 65 are effective by means of silica gel thin layer chromatography, under VerProteus. As a developer of isopropanol-concentrated aqueous

The compound produced according to the invention and its ammonia-chloroform in the ratio of 2: 1: 1 and

Acid addition salts can be parenterally advantageously with ninhydrin as the dye, the presence of the compound

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fertilize IA with Rf of 0.75, IIA with Rf of 0.81, IIIA with Rf of 0.86 and IVA with Rf of 0.92 and a small amount of the unreacted XK-62-2 confirmed. The reaction mixture is then concentrated under reduced pressure and a slightly yellowish residue is obtained, which is a mixture of compounds IA, IIA, IIIA and IVA.

Example 2

Preparation of 2'-N- [L - (-) - a-Hydroxy - ^ - aminobutyryl] - XK-62-2 (Compound I)

The mixture of compounds IA, IIA, IIIA and IVA obtained according to Example 1 is dissolved in 40 ml of aqueous 50% methanol. Then 0.3 ml of acetic acid is added to this solution and the mixture is then subjected to hydrogenolysis in the presence of 250 mg of 5% activated carbon-palladium at room temperature and at atmospheric pressure for 6 hours. By means of silica gel thin-layer chromatography under the conditions described in Example 1, the presence of compounds I with Rf of 0.52, II with Rf of 0.40, III with Rf of 0.31 and IV with Rf of 0.45, in addition to one small amount of the unreacted XK-62-2 confirmed. The catalyst is then filtered and the filtrate is concentrated under reduced pressure. 15 ml of water are dissolved in the residue obtained in order to dissolve the residue and the solution is flowed through a 2.5 cm diameter column with 150 ml of “Amberlite” CG-50 in ammonium form, a product of Rohm and Haas Co., USA calmly. The column is then washed with 200 ml of water. The elution is then carried out with 0.2N aqueous ammonia and the eluate is taken up fractionally in 10 ml portions. As a result, 0.81 g of XK-62-2 is obtained from fractions No. 118-124. Elution is then continued with 0.4N aqueous ammonia and Compound I is eluted in Fractions # 144-174. These fractions are combined and concentrated to dryness under reduced pressure. The result is 1.42 g of compound I in the form of a colorless product. The characteristics of Compound I are as follows. Melting point 135-140 ° C (decomp. At 143 ° C) Specific rotation [a] D2i1 = + 131.6 ° C (C = 0.098, water) IR absorption spectrum (KBr) (cm * 1): 3.700-3.100, 2,950, 1,640, 1,535, 1,490, 1,385, 1,340, 1,287, 1,147, 1,110, 1,055, 1,028, 958, 820, 600 (Fig. 1).

NMR spectrum (in deuterium oxide) S (in ppm from DSS): 1.18 (3H, single »), 2.41 (3H, single), 2.60 (3H, single), 2.83 (2H, single) ), 5.33-4.85 (multiplet overlaps the signal from OH) (Fig. 2).

Elemental analysis for: C2., HlsN „0 ,,. HaCO; 1 Calculated: 'C 47.91 "H 8.04 N 13.41 Found: C 47.97 H 8.27 N 13.51 These results confirm that Compound 1 has the following structural formula:

no.

3rd

nh.

0

nh nh

-ho-

c = 0

ch-oh ch.

high.

nh ch3 oh nh.

Example 3

Preparation of l-N- [L - (-) - a-Hydroxy - "{- aminobutyryl] - XK-62-2 (Compound II)

Following the elution of compound I in example 2, compound II is eluted in fractions No. 225-250. These fractions are combined and evaporated to dryness under reduced pressure, and 0.69 g of compound II is obtained in the form of a colorless product.

The characteristics of this compound II are as follows: Melting point = 120-124 ° C

Specific rotation: [a] D29 = + 99.0 ° (C = 0.10 water)

IR spectrum (KBr) (cm "1): 3,700-3,100, 2,940, 1,640, 1,565, 1,480, 1,385, 1,340, 1,282, 1,111, 1,054, 1,022, 973, 816, 700-600 (Fig. 3).

NMR spectrum (in deuterium oxide) 8 (in ppm from DSS): 1.20 (3H, singlet), 2.36 (3H, singlet), 2.52 (3H, singlet), 5.14 (1H, doublet, J = 4.0 Hz) 5.22 (1H doublet, J = 4.0 Hz) (Fig. 4).

Elemental analysis for: C21H4.sN (; Or). ^ H2C03 Calculated: C 49.39 H 8.29 N 14.11 Found: C 48.96 H 8.37 N 13.95 These results confirm that Compound II has the following structural formula:

no.

o oh nh-c-ch — ch -ch_-nh,

ho ch nh

Example 4

Preparation of 3-N-2'-N-bis- [L - (-) - a-hydroxy-f-amino-butyryl] -XK-62-2 (Compound IV)

Following the elution of compound II, compound IV is eluted in fractions No. 273-291. These

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Fractions are combined and concentrated to dryness under reduced pressure, and 0.53 g of compound IV is obtained in the form of a colorless product.

The characteristics of this compound IV are as follows: Melting point: 157-160 ° C (decomp. At 167 ° C)

Specific rotation: [a] D1'M = + 56.1 ° (C = 0.11, water)

IR spectrum (KBr) (cm "1): 3.700-3.000, 2.930, 1.640, 1.525, 1.470, 1.384, 1.320, 1.140, 1.110, 1.050, 1.020, 953, 870, 815, 600 (Fig. 7).

NMR spectrum (in deuterium oxide) 5 (in ppm from DSS): 1.22 (3H, singlet), 2.36 (3H, singlet), 2.53 (3H, singlet), 5.12 (1H, doublet, J = 4.0 Hz), 5.35 (1H, doublet, J = 3.9 Hz) (Fig. 8).

Elemental analysis for: CasH ^ NfO ^. 3H2C03. H20 Calculated: C 42.81 H 7.52 N 11.28 Found: C 42.79 H 7.30 N 11.20 The results above confirmed that the connection

The analysis of compound III shows the following features:

Melting point: 152-156 ° C (decomp. At 160 ° C)

Specific rotation: [a] D25 = + 91.7 ° (C = 0.11, what-

5 ser)

IR absorption spectrum (KBr) (cm-1): 3,700-3,000, 2,960, 1,650, 1,540, 1,490, 1,387, 1,330,1,150, 1,115, 1,060, 1,023, 960, 820, 600 (Fig. 5).

NMR spectrum (in deuterium oxide) 5 (in ppm from DSS): io 1.22 (3H, singlet), 2.57 (3H, singlet), 2.73 (3H, singlet), 5.40-5.10 (Multiplet overlaps the signal from OH) (Fig. 6). Elemental analysis for: C28H55N701: l. 2H20CO3. H2 Calculated: C 44.61 H 7.56 N 12.14 Found: C 44.86 H 7.29 N 12.31 15 The above results confirm that Compound III has the following structural formula:

2nd

nh ch3 oh

Example 5

Preparation of l-N-2'-N-bis- [L - (-) - a-Hydroxy-f-amino-butyryl] -XK-62-2 (Compound III)

The elution of compound IV in the example above is followed by the elution of compound III with fractions No. 315-341. These fractions are combined and evaporated to dryness under reduced pressure, and 0.72 g of compound III is finally obtained in the form of a colorless product.

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Example 6

Preparation of the monosulfates of compounds I, II, III and IV

1 mol of 2'-N- [L - (- Ja-Hydroxy-y-aminobu-40 tyryl] -XK-62-2 (Compound I) is dissolved in 2 l of water. A solution of 1 Mol of sulfuric acid in 500 ml of water is added with cooling, after 30 minutes cold ethanol is added to the solution to form a precipitate and until the precipitation has ended 45 The precipitate is filtered and the desired monosulfate of compound I is obtained. The monosulfates of Compounds II, III and IV are obtained in the same way.

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2 sheets of drawings

Claims (4)

618180 (1) R4 'Y1Y2N-CH2-CH2-CHOH-CO- and R5' and R0 'are hydrogen, (2) R0 'Y1Y2N-CH2-CHOH-CO- and R4' and R5 'represent hydrogen, 2nd PATENT CLAIM Process for the preparation of a compound of the formula ca) ch3 oh where RG is the grouping oh o ! I! h2n — ch2 — ch2 — ch — c— is or a pharmaceutically acceptable acid addition salt thereof, characterized in that the free 30 amino groups are acylated in a compound of the formula A in which R0 is hydrogen by reaction with a compound of the formula Y, Y2N-CHO-CH.-CHOH-COZ ° 2n n 'N 35 wherein z -rO-n ch -o-c-3 O » H3-Cfl2-C- Cl, Br, I or OH and wherein Yj is hydrogen and Y2 is a protecting group selected Ri JR. O I ch o i 3 n ç,)) - gh2-0-c-, ch3 ~ c-0-c- C * L , c h-o-c- '2 5. Oi- or is, ho2 (3) R4 'and R5' Y ^ N-CH.-CH.-CHOH-CO and RG 'are hydrogen or (4) R "'and Ru' Y1Y2N-CH2-CH2-CHOH-CO- and R, 'is hydrogen, where Yx and Y2 have the above meaning, the protective group Y2 is removed, the compound A is isolated from the mixture, or first of all a mixture from the mixture in which R "is Y, Y2N-CH2-CH, -CHOH-CO- and R4 'and R5' are hydrogen, isolated and thereafter the protecting group Y2 removed, and, if appropriate, a compound A thus obtained is converted into an acid addition salt with a pharmaceutically acceptable acid. 5 The present invention relates to a method for producing a semi-synthetic derivative of the antibiotic XK-62-2, the production of which is described in Swiss Patent No. 568,388. It is apparent from this patent that the anti-io biotic XK-62-2 is obtained by cultivating Actionomycetes such as Micromonospora sagamiensis, Micromonospora echinospora and Micromonospora purpurea by methods commonly used in the cultivation of Actinomycetes. Strains of the microorganisms are inoculated into a liquid medium containing a carbon source such as sugar, hydrocarbons, alcohols, organic acids, etc. inorganic or organic nitrogen sources and inorganic salts and growth promoting agents, and then at 25-40 ° C for 2 Bred for 12 days. XK-62-2 is isolated and purified by combining adsorption and desorption from ion exchange resins and activated carbon, as well as column chromatography using cellulose, «Sephadex» and Sili-cagel. In this way, the antibiotic XK-62-2 can be obtained in the form of the sulfate or in free form. The following published patent applications are known in the prior art, in which compounds of similar constitution are described, but which are not identical to those of the present invention. 30 DT-OS 2 311 524 describes the preparation of l- [L - (-) - Y-amino-a-hydroxybutyryl] -tobramycin from tobramycin, and the Swedish patent application 7 209 211-7 describes the preparation of l- [L - (-) - Y-amino-a-hydroxybutyryl] -kan-amycin A and l- [L - (-) - Y-amino-a-hydroxybutyryl-kanamy-35 a B from kanamycin A or Kanamycin B. XK-62-2 is a basic substance and is obtained as a white powder. It has a molecular formula of C20H41-N-, Oj and a molecular weight of 463. The substance is readily soluble in water and methanol, easily soluble in ethanol 40 and acetone and insoluble in chloroform, benzene, ethyl acetate and n-hexane. According to the present method according to the invention, new antibacterial compounds are produced in such a way that the antibiotic XK-62-2, which has the following formula 45 (B) CII3 Olì 618180 3rd 618180 wherein Ri and R2 are the same or different and are hydrogen, OH, NO2, Cl, Br, I, alkyl having 1-5 carbon atoms or alkoxy having 1-5 carbon atoms and R3 is hydrogen, Cl, Br or I, or Yj and Y2 form a phthaloyl group to form a mixture of compounds of formula (C) p VWCKj. vj "% X0 wherein 4th chemically modified. The derivatives thus produced according to the invention have a strong antibacterial activity against a large number of gram-positive and gram-negative bacteria and in particular they have a remarkably strong antibacterial activity against bacteria which are resistant to known aminoglycoside antibiotics. They can therefore be used for cleaning and sterilizing laboratory equipment and surgical instruments and can also be used in soaps for disinfection purposes and in the cleaning and disinfection of hospital rooms as well as disinfection of rooms used for the production of food. Furthermore, it is to be expected that the compounds produced according to the invention are effective in the treatment of various infections, such as, for example, infections of the urinary and respiratory tract which arise from various bacteria causing inflammation.