DK172543B1 - Process for the preparation of 1-N- (omega-amino-alpha-hydroxyalkanoyl) aminoglucosides and polysilylated aminoglycoside so - Google Patents

Description

in DK PR 172543 B1

The present invention relates to a difficult process for the preparation of 1-N- [w-amino-e-hydroxyalkanoyl] aminoglycosides of the formula

'TTW

R RJ0 I

HO-CH

wherein n is an integer of from 0 to 4; R is a substituted hexo-pyranosyl ring so defined hereinafter; R 3 represents hydrogen or a substituted hexopyranosyl ring so defined as

A

the; R is hydrogen, hydroxy or a pentofuranosyl ring as hereinafter defined; and R 3 represents hydrogen or hydroxy; below the proviso that when R 3 is different from hydrogen, one of the groups 5 and R 5 represents hydrogen and the other hydroxy; and under the assumption that when R 3 represents hydrogen, R 3 represents hydrogen and R * is a substituted pentofuranosyl ring.

The process involves substituting a polysilylated aminoglycoside prepared from an aminoglycoside of formula (XIV): nh, R RO 35 optionally Containing from 1 to 3 aminoblocking groups other than silyl on other amino groups than the Cl amino group, 1 an organic solvent, optionally containing up to approx. 40% water, with an acylating derivative of an acid of the formula: BNS DOCID; <DK 172543B »2 DK PR 172543 B1 B-HN-CH2- (CH2) n-CH-COOH (XIII)

OH

5 wherein B represents an amino-blocking group and n has the meaning described above. All blocking groups are then removed by conventional grace to produce the desired compound of formula (I) · The aminoglycosides are a well-known category of antibiotics and have been widely described in the literature. An excellent review article is the article "Structures and Syntheses of Aminoglycoside Antibiotics" by Suflrio Umezawa 1 "Advances 1n Carbohydrate Chemistry and Biochemistry, JO, 111-182, Academic Press, NY (1974). This review article (and the references cited therein) ) also discusses many known 1N- (acyl) aminoglycoside antibiotics such as * 1N- [L - (-) - 7-amino-0,3 -3 ', 4'-dideoxycanamycin B, tobramycin, paromomycin I, Hbostamydn, 3', 4'-diodeoxyyrbostamycin and lividomycline A.

United States Patent No. 4,029,882 discloses 1-N-acyl derivatives of genta micelles A, B, Bj, Cj, Cja> C2, C2a and X2, slsomldn, verdamln, mutamln 1, 2, 4, 5 and 6 and antibiotics G-418, 66-40B, 66-400, JI-20A, JI-20B and G-52, wherein the acyl groups are derived from a straight chain, branched or cyclic alkyl group containing from 1 to 8 carbon atoms which may contain a amino or hydroxy substituent or both an amino and a hydroxy substituent. the compounds are prepared by acylation of a partially neutralized acid addition salt of the antibody with an acylating derivative of the desired side chain acid.

U.S. Patent No. 4,055,715 discloses the 1N- [L - (-) - 7-amino-e-hydroxy-butyryl] derivative of the aminoglycosyl XK-62-2 and a process for its preparation by acylation of XK-62 -2, if 2'-amino or 2'- and 6'-α * 1no groups are protected by a known amino protecting group (such as the carbobenzyloxy group), with an acylating derivative of L - (-) - t-amino-α-hydroxybutyric acid (such as its N-hydroxy succinic ester).

British Patent No. 1,500,218 discloses the D, L and D, L forms of 1-35 N- [d-amino-o-hydroxypropylone] XK-62-2 and their preparation by substantially the same procedure as described. in U.S. Pat.

4055715.

British Patent No. 1,499,041 discloses 1N- [L - (-) - 7-amino-e-hydroxybutyryl) -6'-N-alkylcanamycin A, wherein the 6'-N-alkyl group of the 1-BNSDOCPD: <DK « 7'Μ3β> 3 DK PR 172543 B1 holds from 1 to 4 carbon atoms. The compounds are prepared, inter alia, by reacting a 6 '-N-alkyl kanamydn A (either unprotected or with the 3 or 3' amino group protected by a conventional amino-blocking group) with an acylating derivative of L - (-) - t -amino-e-5 hydroxybutyric acid.

British Patent No. 1,475,481 discloses 1-N-acyl derivatives of 6'-N-methyl-3 ', 4'-dideoxycanamycin B, wherein the acyl groups may be in the L or D, L form and have the formula:

II

H2N- (CH2) -CH-C- i

OH

Wherein the n is 1, 2 or 3. The compounds are prepared by acylation of the aminoglycoside (whose 6'-amino and optionally 2'-amino group is protected by a conventional amino-blocking group) with an acylating agent containing the above acyl group, e.g. The N * hydroxysuccin1m1 ester thereof.

South African Patent No. 77/1944 discloses, inter alia, a process for the preparation of 1- N- (1-lower) alkanoyl derivatives of kanamydn A and B, wherein the alkanoyl groups may be substituted by hydroxy and / or amino. The method comprises acylation of kanamydn A or B, wherein the 3-amino group of kanamydn A or B and the 2'-amino group of kanamydn B (and optionally the 6'-amino group of the individual antibiotics) are protected by a conventional amino-blocking group. Acylation is achieved in a conventional manner, such as using the N-hydroxy succinyl ester of the acylating acid.

U.S. Patent No. 3,974,137 discloses and Contains claims for a process for the preparation of 1- {L - (-) - 7-amino-o-hydroxybutyryl) -canamydn A, comprising 6'-carbobenzyloxy kanamycin A is reacted with at least three moles of benzaldehyde, a substituted benzaldehyde or plvaldehyde to produce 6'-N-carbobenzyl-oxycanamydn A containing Schlff base groups at the 1-, 3- and 3 "positions, acylation of this tetra-protected kanamydn A derivative with the N-hydroxysuccinimide ester of L - (-) - 7 * benzyloxycarbonylamino or hydroxybutyric acid, and subsequent removal of the protecting groups.

In The Journal of Antibiotics, 790-3 (1973), BNSDOCID refers: <DK 172543B> 4 DK PR 172543 B1 T.P. Culbertson et al. the preparation of 5 "-amino-5" deoxybutirosins A and B from butts in raisins A and B. The first steps of the synthesis included: 1) partial N-trifluoroacetylation of butlrosine base by reflux 5 1 a mixture of methanol and ethyl trifluoroacetate; 2) evaporation to dryness, solution of residue 1 pyridine, treatment with hexamethyldilsilazane and trimethylchlorosilane followed by cooling to less than 10 ° C and treatment with 10 trifluoroacetic anhydride, 3) evaporation to dryness and hydrolyzing the residue in a 2: 1 mixture ethanol and 2N acetic acid under reflux to give tetra [N-trifluoroacetyl)] butirosine.

15

The synthetic end products, 5 "-amino-5" -deoxybutirosins A and B were also reacted in accordance with the above three steps to produce penta [N- (trifluoroacetyl)] - 5 "-amino-5" -deoxybutyrosines A and B. This publication discusses the acylation of a methyl methyl Hylated 20 (and partially acylated) aminoglycoside antibiotic, the result in each case is a complete acylation of all primary amino groups in the molecule (four 1 but1 ros1n starting material) and five. The process of the present invention substantially eliminates polyacylation and provides a high degree of selectivity down to 25 acylation in the desired 1-N position.

J.J. Wright et al. 1, The Journal of Antibiotics, 22 · 714-719 (1976) describes a general method for selective 1-N acylation of the gentamicin-insomnia into the category of aminoglycosides. They report that selectivity for the acylation position is pH dependent and that the 30 C-1 amino group is the most reactive to acylation when the amino group 1 molecule is almost completely protonated. These conditions are obtained by adding an equivalent of a tertiary amine base to a solution of the fully neutralized acid addition salt. Although these researchers achieved 1-N selectivity for the acylation of gentamicin C ^α, 35 slomic acid and foramide, they reported that low selectivity was observed for the acylation of highly hydroxylated aminoglycosides such as gentamicin B and kanamycline A.

British Patent No. 1,460,039 discloses a process for the preparation of various deoxyaminoglycoside antibiotics by halogenation NSDOCID * DK (7? 543B> 5 DK PR 172543 B1) of a phosphorylated aminoglycoside (a glycoside in which the hydroxy group to be removed has been converted to a phosphonoxy group), to produce the corresponding aminoglycoside in which the hydroxy group has been converted to halogen, and reduction of the halogen compound to produce the corresponding deoxyamnoglycoside. Before halogenation of the phosphorylated amylnoglycoside, all of its auxiliary groups are protected. or acyl groups.

The present invention provides a particular process for the preparation of 1'N * [w-amino-α-hydroxyalkanoyl] aminoglycosides. The use of a polysilylated aminoglycoside as a starting material provides high solubility in the organic solvent and thus enables reaction under high concentrations. Although the reaction is usually carried out 1 solutions containing approx. 10-20% polysilylated amino-glycoside starting material, excellent results have been obtained at 15 concentrations of approx. 50% w / v. (for example, 50 g per 100 ml solution).

As with the known processes, the present process provides a mixture of acylated products and the desired product is separated from the other products by chromatography. However, the substitution mode is much more selective in the practice of the present invention, whereby lesser amounts of undesirable products are obtained, which both increases the yield of the desired product and simplifies the chromatographic purification. Thus, in the preparation of 1- (1- (-) - 7-amino-ft-hydroxybutyryl] kanamycin A, amikacin, by various known methods, the 3 "-N-acylated product (ΒΒ-Κ1Ί), the 3 -N-acylated product (BB-K29), the 6'-N-acylated product (BB-K6) and polyacylated material as well as unreacted kanamycln. In commercial scale amikacin production by acylation of 6'-N-carbobenzyloxycanamycin In an aqueous medium followed by removal of the protecting group, it was found that about 10% of the desired amikadn (2.5 kg in a 25 kg 30 charge) was usually lost due to the presence of BB-K11 as a coproduct. The 3 "-N-acylated material produced caused a loss of about an equal amount of the desired 1-N-acylated product due to the great difficulty of separating the latter from the former. The substitution selectivity of the present process is illustrated by the extremely low amount of unwanted 3 "N-acyls t product produced in the preparation of B8-K8 by the present process. Typically, no BB-K11 will be detected in the reaction in the Wed mixture.

The present invention provides a process for BNSDOCiD: <OK 1725438 »6 OK PR 172543 B1 Preparation of 1-N - [« - amino-α-hydroxyalkanoyl] aminoglycosides of Formula (I): • ΤΓΊγ \ λ · R R O C = 0

10 I

HO-CH

CH 2 is NH 2 15 where n is an integer pi from 0 to 4; R represents a hexopyranosyl ring of formula: f6 R6 r6 R10 ^ CHNHR7 In CHNHR7 (II) (III) (IV) wherein R® represents H or CH, R7 represents H or CH, R8 represents OH or NH2, represents H or OH, and R1U represents H or OH; RJ 35 represents H or a hexopyranosyl ring of the formula: M'innniri <Rw 7 DK PR 172543 B1

5 Ho ^ yCH2 ° H PH

h2n ^ -7 \\ r11

I ch3 cow I

10 (v> (vi)

HO \\ - n H.CO ^ K

HN "^ - Λ Λ '—O

15 in «3 ^ or -fsA

(VII) (VIII) wherein R * 1 represents H or CH 3; Represents H or OH, and R4 represents H, OH or a pentofuranosylation of the formula:

“N OH B120 OH

(H) <X 'BNS DOCID <-DK 1725438> 8 DK PR 172543 B1 wherein R1Z represents H or a hexopyranosyl ring of the formula: ch2nh2 "" "γνΙ-O 13 Λν f 5" Λ or R ° ΥΝΙ -O.

(XI) (XII) 'wherein R13 represents H or α-D-mannopyranosyl; provided that when R is different from H, it represents one of the groups R and R H and the other OH; and under the assumption that when R represents H, R H and R represent a pentofuranosylation of formula (IX) or (X) or pharmaceutically acceptable acid addition salts thereof; which is characterized by the addition of a polysyllylated aminoglycoside prepared from an aminoglycoside of formula (XIV): 20 R ° - \ (XIV) R5 R30X \ / NH2

2 3 4 S

wherein R, R, R and R are as defined above, and so may optionally contain from 1 to 3 amino blocking groups, so are different from silyl on amino groups other than the Cl amino group, 1 an organic solvent optionally containing up to 40% water, with an acylating derivative of an acid of formula (XIII): B-HN-CH 2 - (CH 2) n-CH-COOH (XIII)

OH

wherein B represents an amino blocking group and n has the meaning defined above; and then removing all blocking groups.

BM ^ nnrp'v, -ηκ 9 DK PR 172543 B1

The amino groups of the acylating acid of formula (XII) above must be protected with an amino blocking group during the acylation reaction.

This is usually done using a conventional amino-blocking group. These same conventional amino blocking groups 5 can be used to protect amino groups other than the C-1 amino group of the amino glycoside. Such conventional blocking groups for protecting primary amino groups are well known to those skilled in the art. Suitable blocking groups include alkoxycarbonyl groups such as t-butoxycarbonyl and t-amyloxycarbonyl; aralkoxycarbonyl groups such as benzyloxycarbonyl; Cycloalkyloxycarbonyl groups such as cyclohexyloxycarbonyl; haloalkoxy carbonyl groups such as trichloroethoxycarbonyl; acyl groups such as phthaloyl and o-nitrophenoxyacetyl; haloacetyl groups such as trifluoroacetyl; and other well known blocking groups such as the o-nitrophenylthio group, the trityl group, etc.

The acylating acid of formula (XIII) contains an asymmetric carbon atom and may exist in the (+) form or (·) form or as a mixture thereof (d, the 1 form), thus producing the corresponding compound of formula (I) wherein the 1N- [u-amino-α-hydroxyalkanoyl] group is in its (+) [or (R)] form or its (-) (or (S)] form or a mixture thereof.

Acylation of the polysilylated aminoglycosil (with or without from 1 to 3 amino-blocking groups different from silyl on amino groups other than the C1 amino group) can, in general, be carried out in an organic solvent in which the starting material has sufficient solubility of 25 Hg. . These starting materials are highly soluble in most common organic solvents. Suitable solvents include, e.g. acetone, diethyl ketone, methyl-n-propyl ketone, methyl in so-butyl ketone, methyl ethyl ketone, heptane, glyme, diglyme, dioxane, toluene, tetrahydrofuran, cyclohexanone, pyridine, methylene chloride, chloroform and carbon tetrachloride. The choice of solvent depends on the particular starting materials used.

Ketones are generally the preferred solvents. The most advantageous solvent for the particular combination of reactants used can be readily ascertained in routine experiments.

Suitable abrasives for use in preparing the polysilylated aminoglycoside starting materials used herein include abrasives of the formula: BNS DOCID: <DK 172543B> 10 DK PR 172543 B1 r1 \ ^^ r14 f ^ - nh / _15

c 14 'R

5 Ri m ΛΑ I

^ Si / * and R16-Si-Z

R14 ^ n --- Si [n «\ L4 10 R ra k / (XV) (XVI) wherein R * 8, R * 8 and R * 7 are selected from hydrogen, halogen, (lower) al-15 Kyl , (lower) alkoxy, halogen (lower) alkyl and phenyl, At least one of the groups R15, R16 and R17 is different from halogen or hydrogen; R 14 represents (lower) all cooling, m is an integer of 1 to 2, and Z is selected from halogen and R18 20 /

-H

\ "R19

lfi 1Q

Wherein R represents hydrogen or (lower) alkyl and R represents hydrogen, (lower) alkyl or R 15

30 16 I

R - side 17 wherein R 15, R 18 and R 17 have the meaning defined above.

a.NSOOCIO <OK 177M3FI> DK PR 172543 B1 n

Specificsilyl compounds look down formulas (XV) and (XVI) are: trimethylchlorosilane, hexamethyldisilazane, triethylchlorosilane, methyltrichlorosilane, dimethichichlorosilane, triethylbromosilane, tri-n-propylchlorosilane, methyldiethylchlorosilane, methyldiethylchlorosilane, ,, phenyldimethylbromosilane, benzylmethylethylchlorosilane, phenylethylmethylchlorosilane, triphenylchlorosilane, tri-phenylfluorosilane, tri-o-tolylchlorosilane, tri-p-dimethylaminophenylchlorosilanylsilanylsilane, -10 tolyldisilazane, etc. Also suitable are hexaalkylcyclotrisilazanes and octa-alkylcyclotetrasilazanes. Other suitable silylating agents are silyl amides (such as trialkylsilylacetamides and bis-trialkylsilylacetamides), silylureas (such as trimethylsilylurea) and silylurides. Trinethyl silylimidazole can also be used.

A preferred silyl group is the trimethylsilyl group, and preferred silylating agents for introducing the trimethylsilyl group are hexamethylsilazane, bis (trimethylsilyl) acetamide, trimethylsilylacetane and trimethyl chlorine 11 an. Hexamethylenedisilazane is the most preferred.

Polysilylation of aminoglycosides changes the normal order of activity of the amino groups contained therein. Thus, the 6'-amino group is the most active on the kanamycins. If unprotected kanamycin A or B is acylated, the main products are the 6'-N-acylcanamycins. It is for this reason that known methods for preparing 1-N-acylcanamycins required protection of the 6'-N-amino group (e.g. with carbobenzyloxy) to obtain the 1-N-acyl product in good yield. By contrast, when the polysilylated kanamycins are acylated, the main products are the 1-N-acyl kanamycins. This is believed to be due to steric effects of present (or nearby) sllylated hydroxy groups (as well as adjacent glycoside bonds), which prevents acylation by the usually more active amino groups; 30 but this is only a theoretical explanation and does not form part of the invention.

Kanamycin B has the formula: 35 BNS DOCID: <DK 172543S> »-i 12 DK PR 172543 B1 -¾ ApfY · h2N TV / 10 / HO - '° \ / H oN'V' / HO 1/15 ° When kanamycin B with all hydroxy groups sllylyated are considered in the light of the above working theory, it is seen that the 3 "amino group is greatly hindered by the 2" and 4 "silylated hydroxy groups present. It is for this reason that it is not usually detected some 3 "-20 acylated product when the polysilylated kanamycline B (or the structurally similar polysylylated kanamycline A or C) is acylated, although the best 3" N-acylated products are obtained by known methods. Similarly, the 6'-amino group of the neighboring is hindered. 4'and adjacent 3'-silylated hydroxy group The 2'-amino group is hindered by the adjacent 3'-25 silylated hydroxy group and the adjacent glycoside bond.

Other aminoglycosides which are structurally similar to the kanamycins and which, when polysylated, primarily give the 1-N-acyl product, include, e.g. 3'-deoxycanamycin A, 3'-deoxycanamycin B (tobramycin), 6'-N-alkylcanamyclin A compounds, 6'-N-alkylcanamyclin B compounds, 3'-deoxy-6'-N-alkylcanamycline The A compounds, the 3'-deoxy-6'-N-alkyl canamycycline B compounds, the gene A, B, Bj and X2, cell selomycline factors 1 and 3 and the aminoglycosides NK-1001 and NK-1012-1.

Each of these, and other structurally similar aminoglycosides, primarily give the 1-N-substituted product when acylated in the form of their polysyllylated derivative. However, 6'-N- and 3-N-substituted products are formed in small amounts, and one or both of these amino groups may be protected, e.g. with a carbobenzyloxy group.

Another group of aminoglycosides which, although otherwise structurally similar to the kanamycin types described above, contain neither. .nSDOCIO <rDK 17? $ 43B> 13 DK PR 172543 B1 3'- or 4'-hydroxy groups (i.e., 3 ', 4'-dideoxy compounds). When polysyllylated, these do not hinder the sterile 6'-amino group (or 2'-amino group, if present), and 6'-N-substituted (or 2'-6'-di-N-substituted) compounds are the main products of acyl-5 ring. In these aminoglycosides, it is necessary to protect the 6'-amino group (and the 2'-amino group, if present) with an amino-blocking group other than silyl (e.g. with carbobenzyloxy) and acylating the polysylated 6'-N-blocked (or 2 ', 6'-di-N-blocked) aminoglycoside. Aminoglycosides which fall within 10 of this group include e.g. 3 ', 4'-DideoxyCanamycin A, 3', 4'-DideoxyCanamycin B, 6'-N-alkyl-3 ', 4'-DideoxyCanamycin A Compounds, 6'-N-alkyl-3', 4'-DideoxyCanamycin B Compounds , the gentlemen Cj,

Cja> and Cga, aminoglycoside XK-62-2, aminoglycoside 66-400, verdamicin and s1som1cin.

Another category of aminoglycosides are the aminoglycosides in which the glycoside linkage is in the 4 and 5 positions of the deoxystreptamine ring rather than in the 4 and 6 positions. These can be illustrated by ribostamycin having the formula: 20 j. CIUNH-.

HO-X_- ° \ ^ * 7 \\ H2N in * 1 NH, * r * ho, s ·· / "kH '

HO OH

In ribostamycin-type amino glycosides, polysilylation inhibits the desired 1-N-amino group more than the unwanted 3-N-amino group (the other amino groups being hindered as described above for the kanamycin types). Thus, polysilylated ribostamycin-type antibiotics will primarily form the 3-N-substituted product by acylation, and it is therefore necessary to protect the 3-amino group with an amino blocking group as BNS DOCID: <DK 172S43B »14 DK PR 172543 B1 as carbobenzyloxy to obtain the 1-N-substituted product by acylation of the polysilylated starting material. Other aminoglycosides falling within this category include e.g. the neomycins B and C, the paromomycins I and II, the lividomydins A and B, aminoglycosides 2230-C and 5 xylostasin, and their 3'-deoxyders. Also, the 6'-N-alkyl and 3'-deoxy-6'-N-alkyl variants of any of the above-mentioned ribostamycin-type antibiotics containing a 6'-amino group belong to this category. Some of the aminoglycosides of this category contain a 6 'hydroxy group rather than a 6' amino group.

Another group of aminoglycosides are the aminoglycosides of the ribostamycin type described above, which are 3 ', 4'-dideoxy compounds. As with the above-described aminoglycosides of 3 ', 4'-dideoxycanamycin type, the 2'- and 6'-amino groups (in those aminoglycosides of this category containing a 6'-amino group) will not be hindered by polysilylation. Accordingly, for compounds such as 3 ', 4'-dideoxyyrbostaipycin, 3', 4'-dideoxyneomyclines B and C and 3 ', 4'-dideoxyxylostasin and their 6'-N-alkyl analogs, it is necessary to protect 2'-, 3 - and the 6'-amino groups with an amino-blocking group such as carbobenzyloxy, so that acylation of the polysyllylated starting material should primarily produce the 20 IN acyl product. For those aminoglycosides of this category containing a 6'-hydroxy group rather than a 6'-amino group (e.g., 3 ', 4'-dideoxyparomotinyls I and II and the 4'-deoxy1IV1 domycins A or B), it is only necessary to protecting the 2 'and 3-amino groups.

When a starting material is used a polysyllylated amino glycoside containing from 1 to 3 amino blocking groups different from sllyl on amino groups other than the C1 amino group, this starting material can be prepared either by polysilylation of the desired N-blocked aminoglycoside or by introduce the desired N-blocking group 1 the polysyllylated aminoglycoside (if necessary after partial desilylation by hydrolysis or solvolysis).

Methods for introducing silyl groups into organic compounds, including certain aminoglycosides, are known in the art.

The polysyllylated kanamyden (with or without blocking groups different from silyl on amino groups other than the C-1 amino group) can be prepared by methods known per se or as described in this specification.

The term "polysyllylated aminoglycoside" as used herein does not include a persilylated aminoglycoside. Thus, e.g. the term "polysyllylated kanamycline A" kanamycin A containing from 2 OCID: <DK 17? 543B> 15 DK PR 172543 B1 to 10 silyl groups in the molecule [there are a total of 11 positions (4 amino groups and 7 hydroxy groups) which can be silylated].

The precise number of silyl groups present in the polysilylated aminoglycoside starting materials (with or without from 1 to 35 amino-blocking groups different from silyl on amino groups other than the Cl amino group) (or their position ) are not known. It has been found that both lysis and leaching reduce the yield of the desired product and increase the yield of other products.

In case of strong under or overs in lyl is 1ng, a small amount of 10 or None of the desired product can be formed. The degree of silylation which will yield the greatest yield of the desired product will depend upon the many reactants used in the acylation step. The most advantageous degree of silylation using a given combination of reactants can be readily determined by routine experiments.

It is believed that the preferred average number of silyl groups in the polysilylated aminoglycoside starting material will usually be between a lower limit of 4 and an upper limit equal to one more than the total number of hydroxy groups in the aminoglycoside molecule, and that these upper and lower limits are lowered by one for each ami non-blocking group, 20 present in the aminoglycoside molecule. This explanation is merely theory and is not considered part of the invention.

Polysilylated aminoglycosides containing the desired number of silyl groups can be prepared either by using an amount of silylating agent sufficient to add the desired number of silyl groups or by using excess silylating agent to perilylate the aminoglycoside and then desilylate partially. hydrolysis or solvolysis.

Thus, it has e.g. It has been found that in the preparation of 1-N- [L - (-) - γ-amino-o-hydroxybutyryl) kanamycin A by acylation of polysilylated kanamycin A with the N-hydroxysuccin1m1 ester of L - (-) - γ-benzyloxycarbonyl -amino-α-hydroxybutyric acid 1 acetone solution gives good yields of the desired product when using polysilylated kanamycin A, which has been prepared by reacting from ca. 4 to approx. 5.5 moles of hexamethyldisilazane per greater or smaller amounts of hexamethyldisilazane may be used, but the yield of the desired product at the subsequent acylation stage is considerably lowered. In the specific method outlined above, it is preferred to use from ca. 4.5 to approx. 5.0 moles of hexamethyldisilazane per moles of kanamycin in order to obtain maximum yield of product at the acylation step.

BNS DOCID: <DK 172543B> 16 DK PR 172543 B1

It will be appreciated that each mole of hexamethyldisilazane is capable of introducing two equivalents of the trimethylsilyl group 1 kanamydn A or B. Kanamydn A or B each has eleven positions (NH position and OH groups) which can be silylated while kanamydn A and B containing a blocking group which is different from silyl on a different amino group than the C1 amino group, each having a total of such positions. Thus, 5.5 nol of hexamethyldisilazane per theoretically completely silylate all OH and NH 2 groups on the kanamycin, while 5.0 mol of hexamethyldisilazane could completely silylate one mol of kanamydn A or 10 B containing a blocking group different from silyl on another amino group than the Cl amino group. However, it is believed that such extensive silylation does not occur with these molar ratios within reasonable reaction times, although higher rates of silylation are obtained within a given reaction time when a slurry catalyst is added.

Silylation catalysts greatly accelerate the silylation rate. Suitable silylation catalysts are well known in the art and include, inter alia, amine sulfates (which may be the aminoglycosulfate), sulfamic acid, imidazole and tetramethylchlorosilane. Silylation catalysts generally promote a higher degree of silylation than is required for the process of the invention. However, oversilylated ainoglycosides can be used as starting material if first treated with a desilylating agent to reduce the degree of silylation before conducting the acylation reaction.

Thus, e.g. good yields of desired product when acylating polysilylated kanamydn A prepared using a 5.5: 1 molar ratio of hexamethyldisilazane to kanamydn A. However, kanamyclin A silylated with a 7: 1 molar ratio of hexamethyldisilazane (or with a 5, 5: 1 molar ratio in the presence of a silylation catalyst, 1 acetone was acylated with the N-hydroxysuccinimide ester of L - (-) - γ-benzyloxycarbonylamino-e-hydroxyl butyric acid, less than a 1% yield of the desired product was obtained. the same "oversylated" kanamydn A was acylated with the same acylating agent in acetone solution to which water [21 moles water per mole kanamydn; 2.5% water 35 (w / v)] had been added as a desilylating agent 1 hour before the acylation, a yield of about 40% of the desired product was obtained.The same results are obtained if the water is replaced by methanol or another active hydrogen compound capable of desilylation, e.g., ethanol, propanol, butanediol, methyl, mercaptan, ethylmercaptan, BNSDOCIO <OK 17? 543B »17 OK PR 172543 B1 phenylmercaptan or the like.

While it is customary to use terre solvents when working with flushing materials, it has surprisingly been found that adding water to the reaction solvent prior to acylation, 5 even 1 absence of "flushing", often yields equally good yields and sometimes gives better yields of a single product than a dry solvent. Thus, it has e.g. by acylation reactions carried out in acetone at the usual concentrations of 10-20% (w / v) of poly-silylated kanamycin A, it was found that excellent yields of 1N- [L - (-) - 10 Tf-amino-o-hydroxybutyryl ] kanamycin A is obtained by adding up to 28 moles of water per ml. mole polysylated kanamycin A; at a concentration of 20%, 28 moles per mole about 8% water. In other combinations of reactants and solvents, even more water may be tolerated or beneficial. The acylation reaction can be carried out in solvents containing up to approx. 40% water, although at high water concentrations, short acylation times must be used to avoid excessive disilylation of the polysilylated aminoglycoside starting material. A preferred interval is up to approx. 20% water, a more preferred range is up to approx. 8% water and the most preferred range is up to approx. 4% water.

Unless, as described above, solvents containing very large quantities of water are used, the duration of the acylation reaction is not critical. Temperatures can be used in the range of from approx.

-30gC to approx. 100DC in reaction times varying from about 1 hour to up to 1 day or more. The reaction usually proceeds well at room temperature and can be carried out at ambient temperature for convenience. However, for maximum yields and selective acylation, it is preferable to carry out the acylation by from ca. 0 to 5 °.

Acylation of the 1-amino group 1 of the polysilylated aminoglycoside (with or without a blocking group other than sllyl on an amino group other than the C1 amino group) can be carried out with any acylating derivative of the acid of formula (XIII) for the region is known to be suitable for acylation of a primary amino group.

Examples of suitable acylating derivatives of the free acid include the corresponding acid anhydrides, mixed anhydrides, e.g. alkoxy formic anhydrides, acid halides, acid acids, active esters and active thioesters. The free acid can be coupled to the polysilylated aminoglycoside starting material after the free acid is first reacted with N, N'-dimethylchloroforminminum chloride [cf. British Patent Specification No. 1,008,170 BNSDOCID: <DK 172543Β »18 DK PR 172543 B1 and Novak and Weichet," Experientia "XXI, 6, 360 (1965)] or using a Ν, Ν'-carbonyldiimidazole or an N, N'-carbonyld1tr1azole [cf. South African Patent Specification No. 63/2684] or a carbodylmide reagent [ises N, N'-dicyclohexylcarbodiimide, Ν, Ν'-diisopropylcarbodiimide or N-cyclohexyl-N '- (2-morpholinoethyl) carbodiimide Sheehan and Hess, J.A.C.S., 77, 1967 (1955)], or of an alkynylaidine reagent [cf.

R. Buijle and H.G. Viehe, Angew. Chem. International Edition, 3, 582 (1964)] or of an isoxazolium salt reagent [cf. RB Woodward, R.A.

Olofson and H. Mayer, J. Aner. Chem. Soc., 83, 1010 (1961)], or of a ketenimine reagent [cf. C. L. Stevens and M.E. Mund, J. Amer. Chem. Soc., 80, 4065 (1958)] or of hexachlorocyclotriphosphatriazine or hexa-bromocyclotriphosphatriazine (U.S. Patent No. 3,651,050) or of diphenylphosphoryl azide [ODPA; J. Amer. Chem. Soc., 94, 6203-6205 (1972)] or of diethylphosphoryl cyanide [OEPC; Tetrahedron Letters No. 18, pp. 1595-15 1598 (1973)] or of diphenylphosphite [Tetrahedron Letters No. 49, pp.

5047-5050 (1972)]. Another acid equivalent compound is a corresponding azolid, ie. an amide of the corresponding acid whose amide nitrogen is included in a quasiaromatic five-membered ring containing at least two nitrogen atoms, i. imidazole, pyrazole, the triazoles, benzimidazole, benzotriazole and the substituted derivatives thereof. As will be appreciated by those skilled in the art, it may sometimes be desirable or necessary to protect the hydroxyl group on the acylating derivative of the acid of formula (XIII), for example, using acylating derivatives such as an acid halide. Protection of the hydroxy group may be accomplished in ways known in the art, e.g. using a carbobenzyloxy group, by acetylation, by silylation or the like.

In a preferred embodiment of the invention, the acylating derivative of the acid of formula (XIII) is an active ester, and preferably its active ester with N-hydroxysuccinylmide, N-hydroxy-5-norbornene-2,3-30 dicarboxylmide or N-hydroxyphthalimide. In another preferred embodiment, the acylating derivative of the acid of formula (XIII) is a mixed acid anhydride, and preferably its mixed acid anhydride with pivalic acid, benzoic acid, isobutyl carboxylic acid or benzyl carboxylic acid.

After the acylation of the polysilylated aminoglycoside is complete, all blocking groups are removed in 1 known manner to produce the desired product of formula (I). Eg. the silyl groups are easily removed by hydrolysis with water, preferably at low pH.

Also, any amine-blocking groups on the aminoglycoside molecule or on the acyl acid chain can be removed in known manner. Thus, a t-butoxy- * Nsnnrm · n * 179 ^ 7 «-, carbonyl group can be removed using formic acid, a carbobenzyloxy group by catalytic hydrogenation, a 2-hydroxy-1-naphthcarbonyl group by acid hydrolysis. a trichloroethoxycarbonyl group by treatment with zinc stove 1 glacial acetic acid, the phthaloyl group by treatment with hydrazine hydrate 1 ethanol under heating, the trifluoroacetyl group by treatment with NH4 OH, etc.

Preferred amino-blocking groups suitable for protecting amino groups 1 of both the aminoglycosyl molecule and the amino group 1 of the acylating acid of formula (XIII) are the groups of the formulas: 10 R20, __O CH3oO 15 - CH2OC-, CH2-COC- Y2XCC- 'r21> ^ = / CH3

Q

25 M ° * U

o in 11 X3C-C82-0-C- »8

II

o BNS DOCID: <DK 172S43B> 20 DK PR 172543 B1 wherein R20 and R21 are the same or different and each represents H, F,

Cl, Br, NOg, OH, (lower) alkyl or (lower) alkoxy, X represents Cl, Br, F or I, and Y represents H, Cl, Br, F or 1. A particularly preferred amino blocking group for use in the amtnoglycoside molecule is the carbobenzyloxy group. Particularly preferred amino-blocking groups for use in the acylating acid of formula (XIII) are the carbobenzyloxy, trifluoroacetyl and t-butyloxycarbonyl groups.

Some of the compounds of formula (I) contain a double bond 2 (i.e., where the substituent R has the structure IV). These are compounds 10 derived from such amnoglycosides such as sisomicline, verdamicin, G-52, 66-40B and 66-40D. No lateral connections are used, as one skilled in the art understands that any reduction technique which would reduce the double bond should be avoided. Thus, e.g. For example, amino blocking groups which can be removed by hydrolysis or by means of an alkali metal in liquid ammonia are used, so that the reduction of the double bond which would occur by lateral techniques such as catalytic hydrogenolysis is avoided.

Product yields were determined by various methods. After removal of all blocking groups and chromatography on a "CG-50" (NH₂ +) column, the yield could be determined by Isolating the crystalline solid from appropriate fractions or by microbiological analysis (turbidimetric or plate) of appropriate fractions. Another technique used was highly effective liquid chromatography of the unreduced acylation mixture, i.e. the aqueous solution obtained after hydrolysis of the silyl groups and removal of organic solvent but before hydrogenolysis to remove the remaining blocking group (s). This analysis was not a direct analysis for the final product, but for the corresponding N-blocked compounds.

The instrument used was a Waters Associates "ALC / GPC 244" high pressure liquid chromatograph with a Waters Associates absorption detector of 30 model 440 and a 30 cm x 3.9 mm l.d. "μ-Bondapak C-18" column, under the following conditions:

Mobil phase: 25% 2-propanol 75% 0.01 m sodium acetate pH 4.0 Flow rate: 1 ml / min.

Detector: UV at 254 nm

Sensitivity: 0.04 AUFS

Diluent: DMSO

Injected amount: 5 μΐ. NSDOCin · iDK 1> DK PR 172543 B1 21

Concentration: 10mg / ml.

The curve velocity varied, but 2 m1n./2.54 cm was typical. The above conditions provided UV curves with peaks that were easy to quantitatively measure.

5 The results of the above assays are described in the description as HPLC assays.

In order to avoid repetition of complex chemical names, the following abbreviations are sometimes used in the description: 10 AHBA L - (-) - Tr-amino-tt-hydroxysmic acid

BHBA N-carbobenzyloxy derivative of AHBA

HONB N-hydroxy-5-norbornene-2,3-dicarboximide NAE N-hydroxy-5-norbornene-2,3-dicarboxylmide (or BHBA-'ONB ') activated ester of BHBA HONS N-hydroxysuccinylmide SAE N-hydroxysuccinimide activated ester of (or BHBA * ONS ') BHBA DCC dicyclohexylcarbodiimide DCU dicyclohexylurea 20 HHDS hexamethyl isisazane BSA (bis (trimethylsilyl) acetamide HSA trimethyl sllylacetamide TFA tri fluoroacetyl t-BOC tertvc Carbon Corporation for diatomaceous earth.

"Amberi in Tea CG-50" is a trademark of Rohm and Haas Co. for the chromatographic design of a weakly acidic cationic carboxyl polymethacryl type ion exchange resin.

"μ-Bondapak" is a trademark of Waters Associates for a series of highly effective liquid chromatography columns.

All temperatures herein are listed in ° C.

By the terms "(lower) alkyl" and "(lower) alkoxy", as used herein, is meant alkyl or alkoxy groups containing from 1 to 6 carbon atoms.

By the term "pharmaceutically acceptable acid addition salt" of a compound of formula (I) is meant, as used in the specification and claims, a mono-, di-, tri-, tetra- (or higher) salt formed by BNS DOCID: <DK 172543Β B1 interaction between a molecule of a compound of formula (I) and one or more equivalents of a non-toxic, pharmaceutically acceptable acid 1 depending on the particular compound of formula (1). As will be appreciated, an acid addition salt can be formed at each of the amino groups of the molecule, as well as the 1 amino acid glycoside core, as the 1 acyl chain of ideas. Included among these acids are acetic acid, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, nitric acid, hydrobromic acid, ascorbic acid, hydrochloric acid and citric acid, and the other acids commonly used to form salts of amine-containing drugs.

Most of the aminoglycosides used as starting materials

According to the present invention, are known. Any individual aminoglycoside, which is Not 1 and known per se (e.g., a previously described 6'-N-methyl derivative of a known aminoglycoside) can be readily prepared by methods well known in the art, 15 to preparation of analogous analogs.

Compounds of formula (I) prepared by the present invention are active against gram-positive and gram-negative bacteria and are used analogously to other known aminoglycosides. Many of the compounds of formula (I) are 1 and known per se.

According to another aspect of the present invention, polysilylated aminoglycosides of formula (XIV) or polysilylated aminoglycosides of formula (XIV) are provided containing from 1 to 3 amino-blocking groups which are different from sllyl in amino groups other than the Δal group.

The present invention is described in the following, by way of example, which illustrate preferred embodiments.

, ΓϊΟΟΙΠ DK PR 172543 B1 23

Etegffl? El-A

Preparation of 1- [N-fl - [.: -] - [1- (hydroxybutyrylcanoic acid) A. amikacin, by selective acrylic oolyltrimethylsilyl] 6'-N-carbobenzyloxycanamycline A in anhydrous diethyl cyclo-5'6 (15 g, 24.2 mmol) was slurried in 90 ml tmr acetonitrile and heated to reflux under nitrogen atmosphere. Hexamethyldisilazane (17.5 g, 108.48 mmol} was slowly added over 30 minutes and the resulting solution was refluxed for 24 hours. After removal of the solvent in 1 vacuum (40 °) and complete drying under 10 vacuum (10 mm) ), 27.9 g of a white amorphous solid [90.71% calculated as 6'-N-carbobenzyloxycanamycin A (stlyl) g] were obtained.

This solid was dissolved in 150 ml of dry diethyl ketone at 23®. L - {-) - γ-Benzyloxycarbonylamino-e-hydroxybutyric acid N-hydroxy-5-norbornene-2,3-dicarboxylic acid ester (NAE) (11.05 g, 26.67 mmol) dissolved in 100 ml of dry diethyl ketone at 23 ° was slowly added with good stirring over half an hour. The solution was stirred at 23 ° for 78 hours. The yellow clear solution (pH 7.0) was diluted with 100 ml of water. The pH of the mixture was adjusted to 2.8 (3 n HCl) and stirred vigorously at 23 ° for 15 minutes. The aqueous phase was separated and the organic phase was extracted with 50 ml of water at pH 2.8. The combined aqueous fractions were washed with 50 ml of ethyl acetate. The solution was placed in a 500 ml Parr flask together with 5 g of 5% palladium-pi-carbon catalyst (Engelhard) and reduced at 3.4 atm. Hg for 2 hours at 23 °. The mixture was filtered through a layer of "Dicalite" which was then washed with an additional 30 ml of water. The 25 colorless filtrate was concentrated in vacuo (40-45o) to 50 ml. The solution was loaded onto a 5 x 100 cm "CG-50" (NH 2+) ion exchange column. After washing with 1000 ml of water, unreacted kanamycline A, 3- [L - (-) - t-α] 1no-e-hydroxybutyryl] -canamycline A (BB-K29) and amylcadn were eluted with 0.5 n ammonium hydroxide. Polyacyl material was recovered with 3N ammonium hydroxide.

Bioassay, thin layer chromatography and optical rotation were used to record the elution progress. The volume and observed optical rotation for each eluate fraction as well as the weight and percent yield of solid Isolated from each fraction by evaporation to dryness are summarized below: 35 BNS DOCID: <DK 172543Β »24 DK PR 172543 B1

Volume

Material (ml) ® 578 1 wt (g)% yield

Kanamycin A 1000 -> 0.115 0.989 9.15 BB-K29 1750 +0.24 4.37 32.0

Amikacin 2000 +0.31 6.20 47.4

Polyacyls 900 +0.032 0.288 2.0 10

The diethyl ketone layer consumed was shown by highly efficient liquid chromatography to contain an additional 3-5% amikacin.

Oet amikacin (6.20 g) was dissolved in 20 ml of water and diluted with 20 ml of methanol, and 20 ml of isopropanol was added to give crystallization. 6.0 g (45.8%) of crystalline amikacin were obtained.

EteCTWKl l

Preparation of Ι-Ν-ΓΙ-M-Tr-amino-α-hydroxybutyryllanamycin A. amikacin, by selective acylation of oleyftrimethylsilyl kanamycin A.

20 using blocking In situ

A. Polyftrimethylsil vi 1 kanamycin A

Kanamycin A free base (18 g of activity, 37.15 mmol) was suspended in 200 ml of dry acetone / tril and heated to reflux. Hexamethyldislazane 25 (29.8 g, 184.6 mmol) was added over 30 minutes and the mixture was stirred under reflux for 78 hours to give a pale yellow clear solution. Removal of the solvent in vacuo left an amorphous solid residue (43 g, 94%) [calculated as kanamycin A (silyl) 10].

B. L - (-) - γamino-α-hydroxybutyryl 1 kanamycin A

p * (Benzyloxycarbonyloxybenzoic acid (5.56 g, 20.43 mmol) was suspended in 50 ml of dry acetonitrile at 23 ° N, O-β1-trimethylsilyl acetamide (8.4 g, 41.37 mmol) The solution was kept at 23 ° for 30 minutes and then added over a period of 3 hours under vigorous stirring to a solution of poly (trimethylsilyl) kanamycin A (21.5 g, 17.83 m.mol). The compound was stirred for 4 hours, the solvent was removed in vacuo (40 °) and the oily residue dissolved in 50 ml of dry acetone at 23 ° C.

INSDOCID: <DK 172S43B> 25 DK PR 172543 B1 t - (-) - 7-Benzyloxycarbonylamino-e-hydroxybutyric acid N-hydroxy-5-norbornene-2,3-dicarboxylic acid ester (NAE) (8.55 g, 20.63 µl) 1 mol of acetone was added to the above solution over a period of 5 minutes. The mixture was kept at 23 ° C for 78 hours. The solution was diluted 5 with 100 ml of water and the pH (7.0) lowered to 2.5 (6 n HCl). The mixture was placed in a 500 ml Parr flask together with 3 g of 5-palladium-on-carbon catalyst (Engelhard) and reduced at 2.72 atm. Hg for 2 hours at 23 °. The mixture was filtered through a layer of diatomaceous earth which was then washed with 20 ml of water. The combined filtrate and wash water (168 ml) were determined by microbiological analysis against E. coli to contain about 11,400 mcg / ml (19% yield) amlkadn.

Example 3

Preparation of 1-N-fL - (-) - v -amino-tt-hydroxybutyrene kanamycline A.

15 amikacin, by selective acvlerino of polytrimethylsillylcanamycin A

A. Polytrimethylsilvil kanamvcin A

A suspension of 10 g (20.6 mmol) of kanamycin A in 100 ml of dry acetone / tril and 25 ml (119 ml) of 1,1,1,3,3,3-hexamethyldisilazane was refluxed for 72 hours. This gave a clear pale yellow solution. The solution was evaporated to dryness in vacuo at 30-40 ° C. 21.3 g of poly (trimethylslyl) kanamycin A was obtained as a light yellow-brown amorphous powder [85% yield calculated as kanamycin A (silyl) 1Q].

B. 1-N-fL - (-> - γ-amino-tt-hydroxybutyranecanamycin A

T11 a solution of 2.4 g (2.0 mmol) of poly (trimethylsilyl) kanamycin A in 30 ml of dry acetone was slowly added 2.0 mmol of 1- (-) - γ-benzyloxy-carbonylamino-e-hydroxybutyric acid N-hydroxy-5-norbornene-2,3-dicarboxylic acid ester (NAE) in 10 ml of dry acetone at 0-5 ° C. The reaction mixture was stirred at 23 ° C for one week and then evaporated to dryness in vacuo at a bath temperature of 30-40BC. Water (60 ml) was then added to the residue followed by 70 ml of methanol to give a solution. The solution was acidified with 3 n HCl to pH 2.0 and then reduced at 3.40 atm. Hg for 2 hours using 500 mg of 5% palladium-on-carbon catalyst. The material was filtered and combined filtrate and wash was determined by microbiological analysis against E. coli to contain amikacin 1, a yield of 29.4%.

BNS DOCID: <DK 172543B> 26 DK PR 172543 B1

Example 4

Preparation of amikacin by the acylation of PolWtHmethysilyl) S'-N-Cla cane A 1 tetrahydrofuran with the mixed acid anhydrous choice of acid and acid

fiUBA

A. Preparation of mixed anhydrite BHBA (5.066 g, 20.0 mmol), BSA (4.068 g, 20.0 mmol) and triethylamine (2.116 g, 22.0 mmol) were dissolved. 200 ml genus-dried tetrahydrofuran. The solution was refluxed for 1 1/2 1/4 hour and cooled to 10-10BC. Plvaloyl chloride (2.412 g, 20.0 mmol) was added over a period of 2-3 minutes with stirring and stirring was continued for 2 hours at -10 ° C.

The temperature was then allowed to rise to 23 ° C.

B. Acylation of Polytrimethyl Methyl 6 * -H-Cbz Channel A

Poly (trimethylsilyl) 6'-N-Cbz channel A prepared as Example 1 (5.454 g, 4.97 m.mol, calculated as 6'-Cbz channel A (silyl) g) was dissolved in 50 ml dry (molecular sieve). tetrahydrofuran at 23 ° C. Half of the mixed anhydride solution prepared in step A above (10.0 mmol) was added over 20 minutes with stirring and stirring was continued for 7 days.

Water (100 ml) was then added to the reaction mixture and pH (5.4) was adjusted to 2.0 with 3 m H 2 SO 2. Stirring was continued for 1 hour and the solution extracted with ethyl acetate. Polyacylated material began to crystallize so that the reaction mixture was filtered. After drying over 0505, the recovered solid weighed 0.702 g. Extraction of the reaction mixture was continued 1 Total 4 x 75 ml of ethyl acetate, after which excess ethyl acetate was evaporated from the aqueous layer. An aliquot of the aqueous solution was subjected to analysis by HPLC. The resulting curve indicated a 26.4% yield of di-Cbz amikacin.

The aqueous layer was then hydrogenated in a Parr apparatus at 3.40 atm H pressure for 2 hours using 0.5 g of 10% Pd-on-carbon catalyst. The material was filtered and combined filtrate and wash washes against E.coll to contain a 31.2% yield of amikacin. Am1kac1n / BB-K29 ratios around 9-10 / 1; traces of polyacyl and unreacted kana 35 A present.

ftNSnrvrirY 'ΠΚ 17944 ^ «., 27 DK PR 172543 B1

I & sroe] 5

Preparation of Amikacin by Acrylic Acrylic Trimethylsilyl 6'-N-Cbz Can A 1 Acetone with the Mixed BHBA and Isobittyl Carbonic Acid 5A Preparation of Mixed Anhydrous BHBA (1.267 g, 5.0 mmol (MSA) (1.313 g, 10.0 mmol) in 20 ml of genus dried acetone was stirred at 23 ° C and triethylamine (TEA) (0.70 ml, 5.0 mmol) was added. The mixture was refluxed under N 2 atmospheres for 2 1/2 hours. The mixture was cooled at * 20 ° C and 1SO-10 butyl chloroformate (0.751 g, 0.713 ml, 5.50 m.mol) was added. Triethyl-amine hydrochloride immediately began to be excreted. The mixture was stirred for 1 hour at -20 ° C.

b. Asylum ugly Poly (trimethylsilyl) 6'-N-Cbz channel A prepared as Example 1 (6.215g, 4.9mol, calculated as the (silyl) g compound) was dissolved in stirred acetone (20ml) with stirring. at 23 ° C. The solution was cooled to -20 ° C and the cold solution of mixed anhydride from Step A was slowly added over 30 minutes. The reaction mixture was stirred for a further 1½ hours at -20 ° C and then 1 17 hours at 23 ° C. The reaction mixture was then poured into 150 ml of water at 23 ° C with stirring, pH (7.75) was adjusted to 2.5 with 3 n HCl and stirring was continued for 15 minutes. Acetone was then evaporated in vacuo at 40 ° C. An aliquot of the resulting aqueous solution was subjected to analysis by HPLC. The resulting 25 curve showed a 34.33% yield of di-Cbz amikacin.

Most of the aqueous solution was reduced at 3.40 atm. Hg pressure at 23 (C for 3 1/4 hours using 2.0 g Pd / C catalyst. The catalyst was removed by filtration and combined filtrate and wash bags were determined by microbiological analysis against E. coil to give 1n -30 contain a 35.0% yield of amikacin.

Example. $

Preparation of amylcadn by acylation of oolyltrimethylsilyl) 6 * -N-Cbz channel A in anhydrous cyclohexanone 1 varying times 35 A. Poly (trimethylsilyl) 6'-N-Cbz channel A prepared as in Example 1 (2.537 g, 2.0 m .mol, calculated as 6'-N-Cbz channel A (silyl) g) in 300 ml dry cyclohexanone was acylated for 20 hours at 23 ° C with a NAE solution in dry cyclohexanone (10.8 ml of a 0.1944 m .mol / ml solution, 2.10m.mol). Re BNSDOCIO: <DK 172543B> 28 DK PR 172543 B1 the reaction mixture was then added to 150 ml of water with stirring and the pH (5.6) was adjusted to 2.5 with 3 n HCl. The cyclohexanone was evaporated in vacuo at 40 ° C and an aliquot of the remaining aqueous phase was taken for analysis by HPLC.

Most of the aqueous phase was reduced below 3.40 atm. Hg pressure for 3 hours at 23 ° C using 1.0 g of 10% Pd / C catalyst. The catalyst was removed by filtration and combined filtrate and wash bags were analyzed in crobiology for amikacin.

B. A above was repeated except that the acylation was continued for 1 115 hours instead of 20 hours.

Drier 15 HPLC: analysis Microbiology so-called amikacin

UkCbz.amikiClnl Turbid imetr is k Plate

Sales A 49.18% 42.87% 39.16%

Turnover 8 56.17% 55.39% 38.45% 20

Example 1

Preparation of amikacin by acylino of polvitrimethylsilvil 6'-N-Cbz channel A 1 anhydrous tetrahydrofuran 1 varying time 25 A. Example 6A was repeated except that dry tetrahydrofuran was used as a solvent instead of dry cyclohexanone.

B. Example 6B was repeated except that as the solvent 30 I instead of dry cyclohexanone, dry tetrahydrofuran was used.

Yields HPLC Analysis Microbial Nebulizer Amikacin 35 (di-Cbz amikacin) Turbidimetric Plate

Sales A 29.27% 28.34% 28.18%

Turnover B 33.39% 21.52% 28.63% Y / Y

29 DK PR 172543 B1

Example O

Preparation of amikacin by acylation of polytrimethylsilyl) 6'-N-Cbz channel A in anhydrous dioxane for varying time 5 A. Example 6A was repeated except that the acylation was continued in 44 urns using ter dioxane as the solvent.

B. Example 6 B was repeated except that the acylation was continued for 1 18½ hours using ter dioxane as the solvent.

10

Yields HPLC analysis mcrofeialMlilLanalysis amikacin (di-Cbz amikacin 1 Inrbjd.ijieLr1 $ K PI ad? 15

Turnover A 39.18% 43.27% 33.36%

Turnover B 42.82% 22.55% 33.37% EK & eaPfil. g

Preparation of Amikacin by Deviation of Pipeflytrimethylsilvil 6'N-Cbz channel A in anhydrous diethyl ketone at 7S * C

To a stirred solution of poly (trimethylsilyl) 6'-N-Cbz channel A prepared as Example 1 (2.537 g, 2.0 m.mol, calculated as 6'-N-Cbz channel A (silyl) g) in 32 ml of genus-dried diethyl ketone at 75 ° C was added a solution of NAE (10.8 ml with 0.1944 m.mol / ml diethyl ketone, 2.10 m.mol) over 15 minutes. Stirring was continued at 75 ° C for an additional 3 hours, after which the mixture was poured into 150 ml of water. The pH was adjusted to 2.8 with 3N HCl and the diethyl ketone was evaporated in vacuo at 40 ° C.

30 HPLC analysis of an aqueous phase aliquot showed a 39.18% yield of di-Cbz amikacin.

The majority of the aqueous phase was reduced below 3.39 atm. Hz pressure for 3 1/4 hours at 23 ° C using 1.0 g Pd / C catalyst. The catalyst was removed by filtration and combined filtrate and washings were analyzed microbiology for amikacin. Turbidometric analysis showed 27.84% yield and plate analysis showed 28.6% yield.

BNSDOCtD: <DK 172543Β »30 DK PR 172543 B1 iteampi !. lfl

Preparation of Amikacin by Acquisition of Poly (lr1methyl.Sllyl) AttU A ed NAE at 0-5 ° after Tuber Hydrolysis TOd Yaod

5 A. Silvation of kanamycline A using HUDS with THCS SOM

catalyst

Kanamydn A (10 g of purity 97.6%, 20.14 mmol) in 100 ml of screen-dried acetone / ltrl was brought to reflux under nitrogen atmosphere. A mixture of HHOS (22.76 g, 141 moles, 7 moles per mole of kanamydn A) and TMCS 10 (1 ml, 0.856 g, 7.88 m moles) was added to the reflux reaction mixture over the course of 10 minutes. Reflux was continued for 4/4 hours and the mixture was then cooled, concentrated in vacuo to a yellow viscous syrup and dried under high vacuum for 2 hours. The yield of product was 23.8 g (97.9%, calculated as kanamycline A (silyl) 1Q).

B. Acvlerlno

Poly y (tr 1 methyl silyl) kanamydn A (23.8 g, 20.14 m mol) prepared under step A above was dissolved in 250 ml sieve-dried acetone at 23 ° and then cooled to 0-5 °. Water (3.63 mL, 201.4 mole, 10 mole per mole of 20 polysilylated kanamydn A) was added with stirring and the mixture was allowed to stand under moderate vacuum for 30 minutes. NAE (19,133 moles, 0.95 moles per mole of polysilylated kanamydn A) in 108.3 ml of acetone was then added for less than 1 minute. The mixture was stirred at 0-5 ° for 1 hour, diluted with water, pH adjusted to 2.5, and the acetone was then removed in 1 vacuum. The aqueous solution was reduced by 3.40 atm.

H2 ~ pressure at 23 ° for 2½ hours using 2.0 g of 10% Pd / C as catalyst. The reduced reaction mixture was filtered through "Dicalite", concentrated to ca. 100 ml of 1 vacuum at 40 ° and was then kept on a "CG-SO * (NH₂ +) column (6 L resin, 5 x 100 cm). It was washed with 30 water and eluted with 0.6n-1.0n 3n NH 2 OH 60.25% amikacin, 4.37% BB-K6, 4.35% BB-K29, 26.47% kanamycline A and 2.18% polyacyl compounds were obtained.

Example 11 35 Addition of Amikacin by Acrylic Acrylic Methylsilvation 6'N-Cbz Channel A with SAE at 0-5 »

A. Will celebrate off 6 * -N-Cbz kanamycline A

6'-N-Cbz kanamydn A (20.0 g, 32.4 mmol) in 200 ml of sieve-dried ace- 174 Τ4ς3 ^ 31 DK PR 172543 B1 tontril was brought to reflux under nitrogen atmosphere. HMDS (47.3 ni. 226.8 nmol, 7 nmol per mole of 6'-N-Cbz channel A) was added over 10 minutes and reflux continued for 20 hours. The mixture was then cooled, concentrated in vacuo and dried under high vacuum for 25 hours to give 39.1 g of white amorphous solid (95.4% yield, calculated as 6'-N-Cbz channel A (silyl) g).

B. Acylslog

Poly (trimethylslyl) 6'-N-Cbz channel A (39.1 g, 32.4 mmol) prepared under step A above was dissolved in 400 ml of dry acetone with stirring at 23 °. Methanol (6.6 mL, 162 mL, 5 mole per mole of polysylated 6'-N-Cbz channel A) was added and the mixture was stirred at 23 ° for 1 hour under a strong stream of nitrogen. The mixture was cooled to 0-5 ° and a solution of SAE (11.35 g, 32.4 mmol) in 120 ml of precooled dry acetone was added. The mixture was stirred for an additional 3 hours at 0-5 ° and then placed in a 4 ° cold room for 1 week. Water (300 ml) was added, the pH adjusted to 2.0; the mixture was stirred for 1 hour and the acetone was then evaporated in vacuo. The resulting aqueous solution was reduced at 3.67 atm. Hg pressure for 17 hours at 23 ° using 3.0 g of 10% Pd / C as a catalyst. The oil was then filtered through "Dlcalite", concentrated in vacuo to 75-100 ml, loaded onto a "CG-50" (NH 2+) column and eluted with water and 0.6 n NH 2 OH. 52.52% amikacin, 14.5% BB-K29, 19.6% kanamycin A and 1.71% polyacyl compounds were obtained.

Example 12

Preparation of Amikacin by Acupuncture of Polydril Methylsilyl) Channel A with SAE at 0-5 ° after Recirculation with Water A. Silver Leveling of Kanamycin A with TMCS in Acetonitrile Using Tetramethylguanldine as a Reactor

Kanamycin A (4.88 g, 10.07 mmol) was suspended in 100 ml of screen-dried acetonitrile with stirring at 23 °. To the stirred suspension was added tetramethylguanidine (TMG) (16.234 g, 140.98 m.mol, 14 moles per mole kanamycin A). The mixture was heated to reflux and TMCS (15.32 g, 140.98 mole mol, 14 moles per mole kanamycin A) was added over 15 minutes.

A white precipitate of TMG * HCl was formed after ca. half of TMCS had been added. The mixture was cooled to room temperature, concentrated to a sticky residue and dried under high vacuum for 2 hours. The solid was triturated with dry THF (100 ml) and the insoluble TMG * HCl BNS DOCID <DK 172543B> 32 DK PR 172543 B1 was filtered off and washed down 5 x 20 nine portions of THF. Combined filtrate and washings were concentrated in vacuo at 40 ° to a silica residue and dried under high vacuum for 2 hours. 10.64 g of a light cream-colored, silica renan (87.6% yield, calculated as kanamycline 5 A (s1lyl) 10) was obtained.

B. migration

Poly (triethylsillyl) kanamydn A (10.64 g, 10.07 rn.mol) prepared under step A above was dissolved in 110 ml sieve dried acetone with stirring at 23 ° and the solution cooled to 0-5 °. Water (1.81 ml, 100.7 mole, 10 moles per mole of polysilylated channel A) was added and the solution was stirred for 30 minutes under moderate vacuum. SAE (3.70 g, 10.57 n.mol, 5% excess) in 40 ml of precooled dry acetone was added over less than 1 minute and the mixture was stirred for 1 hour. The 81 second was worked up 15 by the general method of Example 11B to produce approx.

50% amikacin, approx. 10% BB-K29, 5-8% BB-K6, approx. 20% kanamydn A and 5-8% polyacyl compound.

Example 13 20 Preparation of Polyethylene Ethylene Polymer A Using Triethylene Chlorosilane with Triethylamine as a Heavy Action Engine

Kanamydn A (5.0 g of 97.6% purity, 10.07 rn.mol) was suspended in 100 ml sieve-dried acetonitrile at 23 *. THethylamine (TEA) (33.8 ml, 24.5 g, 241.7 µmol) was added and the suspension was refluxed.

A solution of trichloroethyl silane (23.7 ml, 21.3 g, 140.98 rn.mol) in 25 ml of dry acetonitrile was added over 20 minutes. Reflux was continued for an additional 7 hours and the mixture was cooled to room temperature, after which long fine needles of TEA * HCl were separated. The mixture was allowed to stand at room temperature for 16 hours, concentrated in vacuo at 40 ° to a sticky solid, and dried for 2 hours under high vacuum to a deep orange sticky solid. The solid was triturated with 100 ml of dry THF at 23 ° and the insoluble TEA * HCl was filtered off, washed with 5 x 20 ml of THF and dried to give 16.0 g of TEA.HC1. Combined filtrate and liquid were concentrated in vacuo to a solid and dried under high vacuum for 2 hours. 19.3 g of poly (triethylslyl) kanamydn A were obtained as a deep orange viscous syrup.

kiWQnnnn ·, nii 33 DK PR 172543 B1

Example 14

Preparation of PolytrimethylBetylSilyl) Kanamydn A Using Bis-Trimethylsilvilurne

Kanamydn A (10.0 g of 99.7% purity, 20.58 mmol) was suspended in 5 200 ml of genus-dried acetonitrile with stirring at 23 °. To the suspension was added bis-trimethylsilurine (BSD) (29.45 g, 144.01 m.mol, 7 moles per mole of kanamydn) and the mixture was brought to reflux under nitrogen atmosphere. Reflux was continued for 17 hours and the reaction mixture was then cooled to room temperature. A small amount of insoluble material was removed by filtration, washed with 3 x 10 ml portions of acetonitrile and dried (1.181 g). Infrared analysis showed that this was BSD plus a small amount of unreacted kanamydn A. Combined filtrate and washes were cooled to 4 ° for 16 hours. Additional solid was separated, removed as above (7.8 g) and shown by infrared analysis to be BSU plus urea. The light yellow filtrate and wash bags were concentrated in vacuo at 40 ° and dried under high vacuum to give 27.0 g of a white solid which was partly sticky and partly fine needle-like crystals. The solid was treated with 150 ml of heptane at 23 °, the insoluble portion removed by filtration, washed with 2 x 20 ml portions of heptane and dried to give 6.0 g of white needles (shown by BSU plus urea) . Combined filtrate and wash bags were concentrated in vacuo at 40 ° and dried under high vacuum for 2 hours to give 20.4 g of white needles, whose infrared spectrum was typical of polysyllylated kanamydn A. Calculations showed that the product contained an average of 7, 22 trimethylsilyl groups.

EKsempd -15

Preparation of Amikacin by Acrylate No Derftrimethylsilyl) Kanamvein A after Partial Desulfurization of 1,3-Butanediol

a. FrewstUling of porttrinftthYlsilyl) can «? yc1n A

Kanamydn A (10.0 g, 20.639 mmol) was suspended in 100 ml of screen-dried acetonitrile with stirring at 23 °. The suspension was refluxed under a stream of nitrogen, and HHOS (23.332 g, 144.5 mmol, 7 moles per mole).

35 moles of kanamydn A) were added over 10 minutes. Reflux was continued for 16 hours and the mixture was then cooled to room temperature, concentrated in 1 vacuum and dried for 2 hours under high vacuum. Crude was obtained 24.3 g of a white sticky residue (92.1% yield calculated as kanamydn A (silyl) n).

BNS DOCID: <DK 172543B> 34 DK PR 172543 B1 B. screening

Per (trimethylsilyl) kanamycline A (24.3 g) prepared under step A above was dissolved in 240 ml of genus-dried acetone with stirring at 23 °.

To this solution was added 1,3-butanediol (9.25 ml, 103.2 m mol, 5 moles per mole per (trimethylsilyl) kanamycin A. The mixture was stirred at 23 ° for 2 hours under a stream of nitrogen and then cooled to SAE (7.23 g, 20.64 mmol) in 70 ml of precooled acetone was added over a period of about 1 minute. The mixture was stirred at 0-5 ° for 3 hours and then allowed to stand for a 4 ° cold 10 room for about 16 hours Water (200 ml) was added, the pH was adjusted to 2.5 and the clear yellow solution was stirred at 23 ° for 30 minutes. The acetone was evaporated in vacuo and the aqueous solution was reduced. at 3.74 atm.

High pressure at 23 ° 1 for 2 hours using 3.0 g of 10% Pd-on-carbon as catalyst (the reduced solution was filtered through "Dlcalite" and chromatographed as Example 11B to give 47.50% amikacin, 5.87% BB-K29, 7.32% BB-K6, 24.26% kanamyclin A, and 7.41% polyacyl compounds.

Etoorool Jfi 20 Preparation of awikadn by acvlerino of polytrimethylsilvil kanamycln A ^ prepared in 1HF using SAE with a solvate catalyst To a reflux mixture of kanamycln A (5.0g, 10.32mol) in 50 ml of hydrochloric acid (THF) was added sulfamic acid (100 mg) and HMDS (12.32 g, 76.33 mmol). The mixture was refluxed for 18 hours, with complete solution occurring after 6 hours. The solution was cooled to 23 °, treated with 0.1 ml of water and kept at 23 ° for 30 minutes. A solution of SAE (3.61 g, 10.3 mmol) in THF 36 ml was added over 30 minutes. After stirring for 3 hours, the mixture was diluted with 100 ml of water and the pH was adjusted to 2.2 with 10% H 2 SO 4. It was stirred for 30 minutes at 23 ° and then concentrated in 1 vacuum to remove THF. The resulting aqueous solution was reduced at 3.40 atm. Hg pressure for 2 hours at 23 ° using 10% Pd-on-carbon as catalyst. The reduced solution was filtered through "Olcalite" and the solid washed with water. Combined filtrate and wash washes (150 ml) were determined by microbiological analysis against E. coli to contain 1225 mcg / ml (31.5% activity yield) of amikacin.

riNsnncin · <ηκ 35 DK PR 172543 B1

CksfiMgl. 11

Prepare Young of amikacin by acylation of polytrimethylsilyl) kanamycin A with the N-hydroxysuccinyl moiety ester of di-carbobenzyloxv AHBA

A. Preparation of di-carbobenzyloxy 1-M-α-amino-o-hydroxybutyric acid N-hydroxysuccinyl ester

Di-carbobenzyloxy L - (-) - tt-amino-α-hydroxybutyric acid (8 g, 20.65 mol) and N-hydroxysuccinimide (2.37 g, 20.65 mmol) were dissolved in 50 ml of dry acetone. at 23 °. Dicyclohexylcarbodiimide (4.25 g, 20.65 mmol) dissolved in 20 ml of dry acetone was added and the whole was stirred at 23 ° for 2 hours. Dicyclohexylurea was filtered off, the filter cake was washed with 10 ml of dry acetone, and the filtrate and washings were combined.

B. Acquisition

Poly (trimethylsilyl) kanamycin A prepared in accordance with the general procedure of Example 15 from 10.0 g (20.639 m.mol) of kanamycin A was dissolved in 100 ml of dry acetone. The solution was cooled to 0-5 °, 3.7 ml of deionized water was added and the solution was stirred at 0-5 ° for 20 minutes under moderate vacuum.

To this solution was added the solution prepared in step A of the di-Cbz-blocked acylating agent and the mixture was stirred at 0-5 ° for 30 minutes. The mixture was diluted with water, the pH adjusted to 2.2 and the acetone removed in vacuo. The aqueous solution was reduced by the general procedure of Example 22 and then filtered through "Dicalite". Chromatography showed 40-45% amikacin, ca. 10% BB-K29, trace amounts of BB-K6, approx. 30% kanamycin A and a small amount of polyacyl compound.

Example 18

Preparation of Dolv (trimethylsilyl) kanamycin A uadec. application of mOS with Imldazole as catalyst

Kanamycin A (11 g, 22.7 m mol) and 100 mg imidazole were heated to reflux in 100 ml of sieve-dried acetonitrile under a stream of nitrogen. HMDS 35 (1B, 48 g, 114.5 mmol, 5 moles per mole kanamycin A was added over 30 minutes and the mixture was refluxed for 20 hours. Complete solution Occurred about 2¼ hours. The solution cooled to 23 ° and the solvent was removed in vacuo leaving 21.6 g of poly (trimethylsilyl) kanamycin A as a foamy residue (93.1% yield calculated as kanamycin BNSDOCIDkOK 172543B> 36 DK PR 172543 B1 (s1lyl) n).

Example 1.9

Preparation of 1-N-rL-M-T-amino-o-hydroxybutyryl kanamyclin B 5 (1BB-K26) by acrylication of polytrimethylsilyl kanamylene 8 with SAE

A. Preparation of Poly (Trinethylsilyl) Kanawvdn B Using HMDS with TMCS Catalyst

Kanamydn B (25 g, 51.7 mmol) in 250 ml of genus-dried acetonitrile is heated to reflux under a nitrogen stream. HHOS (62.3 g, 385.81 m mole, 7.5 moles per mole of kanalycin B) was added over a period of 30 minutes followed by 1 ml of TMCS as catalyst. The mixture was refluxed for 21 hours with complete solution after 1 hour. The solvent was removed in vacuo at 60 ° and the oily residue was maintained at 60 ° under high vacuum for 15 hours. Oops were obtained 53.0 g of poly (trimethylslyl) kanamycline B (85.2% yield calculated as kanamydn B (sllyl) µg).

B. Acceleration

The poly (trimethylsilyl) 20 kanamydn B (53.0 g) prepared in step A above was dissolved in 1,500 ml of dry acetone at 0-5 °, methanol (20.9 ml) was added and the mixture was stirred in vacuo for 30 minutes at 0 ° C. -5 °. A solution of SAE (18.1 g, 51.67 mmol) in 200 ml of cold acetone was added over less than 1 minute and the mixture was stirred for 30 minutes at 0-5 °. The mixture was worked up according to the general procedure of Example 11 and then loaded onto a column of "CG-50" (NH 2+) (8 x 120 cm). It was eluted with an NH 1 OH gradient from 0.6 n to 11 3 n. 38% BB-K26, 5% of the corresponding 6'-N-acylated kanamycline B (BB-K22), 10% of the corresponding 3 were obtained. -N-acylated kanamycline B (BB-K46) 14.63% kanamycline 8 and an Ulle amount of polyacylated kanamycline B

30

Example 20

Preparation of Powder (Trimethylsilyl) Canvane A Using HHOS with Canvane A Sulfate as Catalyst

Kanamycline A (19.5 g, 40.246 mmol) and kanamydn A sulfate (0.5 g, 0.858 mmol) [total - 20.0 g, 41.0 mmol] in 200 ml of genus dried acetonitrile brought to reflux. HMDS (60.3 mL, 287.7 mL, 7 moles per mole kanamycline A) was slowly added and the mixture was refluxed for 28 hours.

It was then evaporated to dryness in a rotary evaporator and dried under a steam injector vacuum. 47.5 g of poly (trimethylslyl) were obtained

-Nsnomn «ίίκ 17? S4:? R

37 DK PR 172543 B1

kanamycline A as a pale yellow oil (95.82% yield calculated as kanamycin A

(Silyl) 10) ·

Example 21 Preparation of Amikacin by Acolylation of Ooly (Trifroethylsilyl) kanamyclin A with N-trifluoroacetyl-bleached AHBA N-hydroxysuccinylmester ester A. Preparation of N-trifluoroacetyl AHBA and conversion to its N-

Mn> xysutt1n.1iWestgr 10 To a suspension of AHBA (5.0 g, 42 m.mol) in 100 ml of THF was added trifluoroacetic anhydride (40g, 191 m.mol) with stirring over 10 minutes. The solution was stirred for 18 hours at 23 ° and then concentrated to dryness in vacuo at 50 °. The residue was dissolved in 100 ml of aqueous methanol (1: 1) and stirred for 1 hour. It was then concentrated to dryness in vacuo and redissolved in 50 ml of H 2 O. The aqueous solution was extracted with 3 x 50 ml portions of MIBK and the extract was concentrated after drying over Na 2 SO 4 to an oil. Solvent traces were removed by addition and distillation of 4 ml of water. On standing, the oil is changed to a waxy crystalline solid (2.5 g, 28% yield).

20 N-trifluoroacetyl AHBA (2.4 g, 11.3 mmol) was dissolved in 50 ml of dry acetone and N-hydroxysuccin1m1d (1.30 g, 11.31 mmol) was added to the solution. A solution of dicyclohexylcarbod11mide (2.33 g) in 20 ml of dry acetone was added slowly. The reaction mixture was stirred for 2 hours at 23 ° and the precipitated dicyclohexylurea was removed by filtration and washed with a small portion of acetone. Combined filtrate and washing liquids (a solution of the N-hydroxy succinyl ester of N-trifluoroacetyl AHBA) were used during the next step without isolation.

B. Acquisition 30 To a solution of poly (trimethylsilyl) kanamycline A prepared as in Example 20. (11.31 moles) in 54 ml of acetone was added 2.0 ml (113.4 moles) of water and the mixture was stirred. in vacuo at 0-5 * for 30 minutes. The N-hydroxysuccinyl ester of N-trifluoroacetyl AHBA prepared under step A above (11.31 m.mol) was added to the mixture and it was then maintained at 5 ° for 1 hour. The pH value was adjusted to approx. 2.0 by 20%

HgSO4, the mixture was stirred for 30 minutes, and the pH was raised to ca. 6.0 with NH 3 OH. The mixture was then evaporated to dryness in a rotary evaporator to give 14.4 g of a clear off-white solid. The solid was dissolved in 100 ml of water, the pH was calculated from BNSDOC1D: <DK 172543B> 38 DK PR 172543 B1 5.5 to 11.0 with 10 n NH 3 OH, and the solution was heated in an oil bath at 70 · 1 L hour. The pH (9.5) was flushed to 7.0 with HCl, the solution fine filtered to remove a small amount of insoluble constituents and the filter washed with water. Combined filtrate and washes (188 ml) were added to a "CG-50" (NH NH + J column (8 x 90 cm)), washed with 2 liters of water and eluted with an NH NHOH gradient (0.6n-1). , 0n-concentrated.) 28.9% amikacin, 5.OX BB-K6, 5.7% BB-K29, 43.8% kanamydn A, 3.25% polyacyl compound plus 14.3% of an unknown material which was 1 the first fraction from the column.

10

Example 22

Preparation of Amikacin by Acrylic Acid Polymer-Methylsilyl) Kanamvenn A with t-Butyloxycarbonyl-Bleached AHBA N-Hydroxysuccinimide Ester 15 A. Preparation of t-BOC AHBA and Conversion to Its N-Hydroxysuccinyl Esters

A solution of AHBA (5.0 g, 42 mmol) in 100 ml of water and 20 ml of acetone was adjusted to pH 10 with 10 n NaOH. Over 3-4 minutes, 11.6 g (53 mmol) of dl-t-butyl dicarbonate was added and the solution was stirred for 20 minutes, maintaining the pH value of 10 for periodic counting of 10 n NaOH. The acetone was removed in vacuo and the aqueous phase was washed with 40 ml of ethyl acetate. The pH of the aqueous solution was lowered to 2.0 with 3 n HCl and the solution was then extracted with 3 x 30 ml MIBK. Oe combined MIBK extracts were dried over NagSO 4 and concentrated to give a clear oily residue (8.2 g, 89X).

-t-BOC-AHBA (4.25 g, 19.4 mmol) was dissolved in 50 ml of acetone and N-hydroxysuccinylmide (2.23 g, 19.4 mmol) was added. A solution of dicyclohexylcarbodime (4.00 g, 19.4 mmol) in 20 ml of acetone was added slowly and the mixture was stirred for 2 hours at 23 °. The unfolded di-cyclohexylurea was removed by filtration and washed with a small amount of acetone. Combined filtrate and washings (a solution of the N-hydroxysuccinyl ester of -t-BOC-AHBA) were used in the next step without Isolation.

B. Acylation

To a solution of poly (trimethylsilyl) kanamydn A prepared as in Example 20 (41.28 mmol) in 94 ml of acetone was added 3.5 ml (194 mmol) water and the mixture was stirred in vacuum at 0-5 °. for 30 minutes. The N-hydroxysuccinyl ester of -t-BOC-AHBA prepared under step A above BNS DOCID: <DK 172S43B> 39 DK PR 172543 B1 (19.4 mmol) was added and the mixture was allowed to stand at 5 ° for 1 hour. Water (200 ml) was added and pH (7.0) was lowered to 2.0 with 20% HgSO 4. After stirring for 30 minutes, the pH was raised to ca. 6.0 with NH 2 OH and the mixture was evaporated to dryness in vacuo to give 36.3 g of a golden oil.

6 The oil was dissolved in 200 ml of trifluoroacetic acid, allowed to stand for 15 minutes and evaporated to dryness in a rotary evaporator. The oil was washed with water and the water flash evaporated. Concentrated NH 4 OH was added to pH 6.0 and flash evaporated. The resulting solid was dissolved in water, filtered and the filter washed with water. Combined filtrate and washes 10 (259 ml) were charged to an aCG-50 "(NH 4 +) column (8 x 92 cm), washed with 4 liters of water and eluted with an NH 2 OH gradient (0.6n-1.0n concentrated).

Oer was obtained 40.32% amylkadn, 4.58% BB-K6, 8.32% BB-K29, 30.50% kanamycin A and 7.43% polyacyl compound.

Example 23

The general procedure of Example 10 is repeated except that the kanamycin A used therein is replaced with a zquimolcr amount of: 3'-deoxycanamycin A, 6'-N-methylkanamycin A, 3'-deoxy-6'-N-methylkanamycin, respectively. A kanamycin B, 6'-N-methylkanamycin B, tobramycin (3'-deoxycanamycin B), 6'-N-methyltobramycin, aminoglycosyl NK-1001, 3'-deoxy-aminoglycoside NK-1001, 6'-N-methyl -amylnoglycosyl NK-1001, 3'-deoxy-6'-N-methyl-aminoglycosyl NK-1001, gentamicin A, 3'-deoxygentam1c1n A, gentamicin 8, 3'-deoxygentamyclin B, 6'-N-methylgentam1c1n B, 3'-deoxy-6'-N-methylgentamicin Bj, gentamicin 8j, 3'-deoxygentamidn Bj, 6'-N-methylgentamicin Bj, 3 '-deoxy-6'-N-methylgentamicin Bj, BNSDOCID: <DK 172S43B> 40 DK PR 172543 B1 gentamicin X2, seldomycln factor 1 and seldomydn factor 2, 5, thereby producing respectively: 1-N-IL - (-) - γ-amino-α-hydroxybutyryl] -3'-deoxycanamycin A, 1- Ν- [L - (-) - γ-amino-α-hydroxybutyryl] -6'-N-methylkanamycin A, 1-N- [1- (-) - 7-amino-α-hydroxybutyryl] -3'-deox γ-6'-H-methyl-kanamycin A, 10N- [L - (-) - γ-amino-α-hydroxybutyryl] kanamycin B, 1-N- [L - (-) - γ-amino-α-α hydroxybutyryl] -6'-N-methylcanamycin B, 1-N- [L - (-) - γ-amino-α-hydroxybutyryl] tobramycin, 1N-1L-1β-amino-α-hydroxybutyryl-e'-N-methyltobramycin , 1N- [L - (-) - 7-Amino-α-hydroxybutyryl] aminoglyco $ 1d NK-1001, 15N- [L - (-) - 7-Amino-α-amino-α-hydroxybutyryl] -3 ' -deoxy-aminoglycoside NK-1001, 1-N- [L- (-) -γ-amino-α-hydroxybutyryl] -6'-N-methyl-aminoglycoside NK-1001, 1-N- [L- (-) -y-amino-α-hydroxybutyryl 1 -3 '-deoxy-6' -N-methyl-aminoglycoside NK 1001, 20N- [L - (-) - 7-amino-e-hydroxybutyryl] gentam1c1n A, 1N- [L - (-) - 7-Amino-e-hydroxybutyryl] -3'-deoxygenetamycin A, 1-N- [L - (-) - 7-Amino-α-hydroxybutyryl] gentam1c1η B, 1-N - [L - (-) - 7-Amino-α-hydroxybutyryl] -3'-deoxygentamine B, 1N- [L - (-) - 7-amino-α-hydroxybutyryl] -6'-N-methylgenta-1c1n B, 1 -N- [L- (-) -7-amino-α-hydroxybutyryl] -3 '-deoxy-6' -N-methyl gentamicin Bj, 1N- [1- (-) - 7-amino-α-hydroxybutyryl ] gentam1c1n 8j, 1-N- [L- (-) - 7-amino- α-hydroxybutyryl] -3'-deoxygentamycin B j, 1N- [L - (-) - 7-amino-α-hydroxybutyryl] -6'-N-methylgentamycin Bj, 1-N- [L- (-) -7 - amino-α-hydroxybutyryl) -3 '-deoxy-6'-N-methyl gentamicin N Bj, 30 lN- [L - (-) - 7-amino-a-hydroxybutyryl] gentamicin X, lN- [L - (-) - 7-Amino-α-hydroxybutyryl] celeromycin factor 1 and 1N- [L - (-) - 7-amino-α-hydroxybutyryl] celeromycline factor 2.

Reaction of each of the above-mentioned aminoglycosid starting materials in the same manner with L - (-) - 0-benzyloxycarbonylamide-no-α-hydroxypropionic acid N-hydroxy-5-norbornene-2,3-dcarboxylmester ester in place of L- The (-) - 7-benzyloxycarbonylamino-α-hydroxybutyric acid N-hydroxy-5-norbornene-2,3-dicarboxylic acid ester produces the corresponding 1N- [L - (-) - β-amino-α-hydroxypropylonyl] aminoglycos in there.

BNS DOCID: <DK <79S43B> 41 DK PR 172543 B1

Reaction of each of the above-mentioned aminoglycoside starting materials in the same manner with L - (-) - 6-benzyloxycarbonylamino-hydroxy-valeric acid-N-hydroxy-5-norbornene-2,3 * dicarboximide ester in place of L- The (-) - 7-benzyloxycarbonylamino-α-hydroxybutyric acid N-hydroxy-5-norbornene-2,3-dicarboximide ester produces the corresponding 1N- [L - (-) - i-amino-ar-hydroxyvaleryl] amine noglycos in the .

Example 24

The general procedure of Example 10 is repeated except that the L - (-) - 7-benzyloxycarbonylanino-t-hydroxybutyric acid-N-hydroxy-5-norbornene-2,3-dicarboximide is used, respectively, with L- ( -) - O-Benzyloxycarbonylamino-α-hydroxypropionic acid N-hydroxy-5-norbornene-2,3-dicarboximide ester and L - (-) - S-benzyloxycarbonylamino-α-hydroxy-valeranoic acid N-hydroxy-5-norbornene-2 , 3 * dicarboximide ester, thereby producing, respectively, 1N- [L - (-) - 0-amino-ε-hydroxypropionyl] kananamycin A and 1-N- [L - (-) - 6-amino-α-hydroxyvaleryl] kanamycin A .

Example 25 The general procedure of Example 1 is repeated except that the 6'-N-carbobenzyloxycanamycin A used therein is replaced by an equimolar amount of 6'-N-carbobenzyloxy-3 ', 4'-dideoxycanamycin A, 25, respectively. 6'-N-carbobenzyloxy-3 ', 4'-dideoxy-6'-N-methylkanamycin A, 2', 6'-di- (N-carbobenzyloxy) -3 ', 4'-dideoxychanamycin B, 2', 6 '-di - (N-carbobenzyloxy) -3', 4'-dideoxy-6'-M-methylcanamycin B, 2 ', 6'-di- (N-carbobenzyloxy) gentamicin C 2, 2', 6'-di (N-carbobenzyloxy) gentamicin Cja, 2 ', 6'-di- (N-carbobenzyloxy) -6'-N-methylgentamicin Cja, 2', 6'-di- (N-carbobenzyloxy) gentamicin C2, 2 ' , 6'-di- (N-carboberzyloxy) -6, -N-methylgentamicin and 2 ', 6'-di - (N-carbobenzyloxy) ami noglycoside XK-62-2, thereby producing 35 l * N * respectively. [1- (-) - 7 * ai »1no-α-hydroxybutyryl] -3 ', 4'-dideoxycanafnycin A,] -N-11 - (-) - 7-amino -' - hydroxybutyryl] -3 ', 4 '-dideoxy-6'-N-methylkanamycin A, 1N- [1- (-) - 7-amino-α-hydroxybutyryl] -3', 4'-dideoxycanamycin B, 1-N- (L- (-) - 7-araino-o-hydroxybutyryl] -3 ', 4'-d ideoxy-6'-N-methylkanamycin BNSDOCID: <DK 172543B> 42 DK PR 172543 B1 B, 1-N- [L - (-) - 7-amino-o-hydroxybutyryl] gentamic1 C, 1-N-IL- ( -) - γ-Amino-o-hydroxybutyryljgentamici n Cj, 1 -N- (L - (-) - γ-amino-o-hydroxybutyryl] -6 '-N-methylgentamicin Cja, 5lN- [l - (- ) -Y-amino-e-hydroxybutyryl] gentamicin C2,1- [1- (-) - T-amino-o-hydroxybutyryl] -6'-N-methylgentamycin C2 and 1-N- [L - (-) - γ-amino-o-hydroxybutyryl] aminoglycoside XK-62-2.

Reaction of each of the above-mentioned aminoglycoside starting materials in the same manner with L - (-) - 0-benzyloxycarbonylamino-α-hydroxy-propionic acid-N-hydroxy-5-norbornene-2,3-dicarboximide ester instead of L The (-) - 7-benzyloxycarbonylamino-o-hydroxybutyric acid-N-hydroxy-S-norbornene-2,3-dicarboxylic acid ester produces the corresponding 1N- [t - (-) - / J-amino-α-hydroxypropionyl] aminoglycosides.

Mixing each of the above-mentioned aminoglycosyl starting materials in the same manner with L - (-) - i-benzyloxycarbonylamino-o-hydroxy-valeric acid-N-hydroxy-5-norbornene-2,3-dicarboxylic acid ester in place of L - The (-) - 7-benzyloxycarbonylamino-α-hydroxybutyric acid-N-hydroxy-5-norbornene-2,3-dicarboximide ester produces the corresponding 1- [1- (-) - 6-amino-α-hydroxyvaleryl] aminoglycosides.

Example 26 2 ', 6'-di- (N-trifluoroacetyl) silica is suspended in dry acetone / ltrl and heated to reflux under nitrogen atmosphere. Hexamethyldisilazane [4 moles 25 pr. moles of 2 ', 6'-di- (N-trifluoroacetyl) silsomcin] are added over 30 minutes and the solution obtained is refluxed for 24 hours. Removal of the solvent in vacuo gives solid polysilylated 2 ', 6'-di- (N-trifluoroacetyl) sisomicin.

The polysilylated 2 ', 6'-d1- (N-trifluoroacetyl) silicon is acylated with the N-hydroxysuccinimide ester of L - (-) - 7-trifluoroacetylamino-o-hydroxybutyric acid in accordance with the general procedure of Example 21B In Example 218 to produce 1-β- [ί - (-) - γ-amino-β-hydroxybutyryl] si somic In.

EKseinpgl 27

The general procedure of Example 26 is repeated except that the 2 ', 6'-d1- (N-trifluoroacetyl) sisom1c1n used therein is replaced by an equimolar amount of -r.SDOCID: <DK 172543B> DK PR 172543 B1 43 2 ', 6'-di - (N-trifluoroacetyl) -5-episisomicin, 2', 6'-di- (N-trifluoroacetyl) -6'-N-methylsisomicin, 2 ', 6'-di - (N-trifluoroacetyl ) -6'-N-methyl-5-epsissomicin, 2 ', 6'-di - (M-tri-fluoroacetyl) verdamicin, 2', 6 '- di - (N-tri-fluoroacetyl) -5-epi verdamin, 2 ', 6'-di - (N-trifluoroacetyl) -6'-N-methyl Verdainicin, 2', 6'-di- (N-trifluoroacetyl) -6'-N-methyl-5 epidemicamine and 2 ', 6'-di - (N-trifluoroacetyl) aminoglycoside 66-40D, 10 thereby producing respectively 1N- (L - (-) - Y-amino-o-hydroxybutyryl] -5-episisomicin, 1 -N- [L - (-) - γ-amino-ε-hydroxybutyryl] -6'-N-methylsomycin, 1-N- (L - (-) - γ-amino-or-hydroxybutyryl) -6'- N-methyl-5-epi-somicline, 1N- [1- (-) - γ-amino-α-hydroxybutyryl] verdamicin, IN- (L - (-) - γ-amino-α-hydroxybutyryl] -5- epiverdamycin, 1-N- [L - (-) - γ-amino-ε-hydroxybu tyryl] -6'-N-methylverdamicin, 1N- [L - (-) - t-amino-α-hydroxybutyryl] -6'-N-methyl-5-epiverdamicin and 1-N- [L - (-) - γ-ami no-α-hydroxybutyryl] am i noglycos id 66-40D.

20

Reaction of each of the above 2 ', 6'-di- (N-trifluoroacetyl) aminoglycoside starting materials in the same manner with the N-hydroxy-succinyl ester of L - (-) - ^ - trifluoroacetylanino-α-hydroxypropionic acid instead for the N-hydroxysuccin1m1 ester of L - (-) - t-trifluoroacetylamino-α-hydroxybutyric acid produces the corresponding 1N- [L - (-) - [(amino-α-hydroxy-propionyl) aminoglycosides).

Mixing each of the above 2 ', 6'-di- (N-trifluoroacetyl) aminoglycoside starting materials in the same manner with the N-hydroxy-succinic ester of 1- (-) - 6-trifluoroacetylamino-e-hydroxyvaleric acid 30 the site of the N-hydroxy succinic midester of 1- (-) - 7-trifluoroacetylamino-o-hydroxybutyric acid produces the corresponding 1 N- [L - (-) - 6-amino-o-hydroxyvaleryl] amine noglycosides.

gtefflQBgl-Zfl 35 The general procedure of Example 26 is repeated except that the N-hydroxysuccinimide ester of L - (-) - γ-trifluoroacetylamino-e-hydroxy-butyric acid is replaced by an equimolar amount of the N-hydroxysuccinimide esters of <17 > 44 DK PR 172543 B1 L - (-) - β-trifluoroacetylamino-α-hydroxypropionic acid and L - (-) - 4-trifluoroacetylamino-α-hydroxyvaleric acid, thereby producing 5 lN- [L - (-) - 0 -amino-α-hydroxypropionyl] silsomycline and 1-N- [L - (-) - 4-amino-t-hydroxyvaleryl] silsomycline.

Example 29 The general procedure of Example 1 is repeated except that the 6'-N-carbobenzyloxycanamycin A used therein is replaced by a quantum amount of 3-N-carbobenzyloxyribostamyclin, 3-N-carbobenzyloxy-3'-deoxy, respectively. 3-N-carbobenzyloxy-6'-N-methylrlbostamydine, 3 * N-carbobenzyloxy-6'-H-methyl-3'-deoxyyrbostamycin, 3-N-carbobenzyloxyneomycline B, 3-Nc arbobenzyloxy-3'-deoxyneomycl 3-N-carbobenzyloxy-6'-N-methylneomycin B, 3-N-carbobenzyloxy-6'-N-methyl-3'-deoxyneomycl π B, 3-N-carbobenzyloxyneomycin C, 3-N-carbobenzyloxy-3 ' -deoxyneomycin C, 3-N-carbobenzyloxy-6'-N-methylneomycin C, 3-N-carbobenzyloxy-6'-N-methyl-3'-deoxyneomydn C, 3-Nc arbobenzyloxyxylosin, 3-N-carbobenzyloxy 3'-deoxyxylostazine, 3-N-carbobenzyloxy-6'-N-methylxylostazine, 3-N-carbobenzyloxy-6'-N-methyl-3'-deoxyxylostazine, 3-N-carbobenzyloxyparomomydine I, 3-N-carbobenzyloxy 3'-deoxyparomomycin I, 2 ', 3-di- (N-carbobenzyloxy) -3', 4'-ddedexyparomomycin I, 3-N-carbobenzylo xyparomomycin II, 3-N-carbobenzyloxy-3'-deoxyparomomydine II, 2β-di- (N-carbobenzyloxy) -3 ', 4'-dideoxyparomomydine II, 3-N-carbobenzyloxy-aminoglycoside 2230-C, 3- N-carbobenzyloxy-3'-deoxy-aminoglycoside 2230-C, 3-N-carbobenzyloxyl ivldomydn A and 3-N-carbobenzyloxylivldomycin B ,. NSDOCID: <DK 1725438> 45 DK PR 172543 B1, thereby producing 1-N- [1- (-) - 7-amino-α-hydroxybutyryl] ribostamycin, 5 lN- [L - (-) - 7-amino -e-hydroxybutyryl] -3'-deoxyyrbostamycin, 1-N- [L - {-) - 7-amo-no-o-hydroxybutyryl] * 6'-N-methyl ribostamycin, 1N- [L - (-) - 7 -amino-α-hydroxybutyryl] -6'-N- * ethyl-3'-deoxyribostamycin, 1-N- [L - (-) - 7-amino-o-hydroxybutyryl] neomyc in η B, lN- [l - (-) - 7-Amino-o-hydroxybutyryl] -3'-deoxyneomyciη B, 10 N- [L - (-) - 7-Amino-a-hydroxybutyryl] -6'-N-methylneomycline B, 1-N - [1- (-) - 7-Amino-o-hydroxybutyryl] -6'-N-methyl-3'-deoxyneomycline B, 1N- [L - (-) - 7-amino-o-hydroxybutyryl] neomycin c, 1-N- [L - (-) - 7-amino-o-hydroxybutyryl] -3-deoxyneomycin C, 1-N- [L - (-) - 7-amino-o-hydroxybutyryl] -6'-N -methyl neomyc in C, 15 NN- [L - (-) - 7-amino-o-hydroxybutyryl] -6'-N-methyl -3'-deoxyneomycin C, 1-N- [L - (-) - 7 -amino-o-hydroxybutyryl] xylostasin, 1-N- [L - (-) - 7-amino] -α-hydroxybutyryl] -3-deoxyxylostazine, 1-N- [L - (-) - 7-amino-o- hydroxybutyrylJ-6'-N-methylxylostasylin, 1N- [L - (-) - 7-amino] -o-hydroxybutyryl1-6'-N-methyl-3'-deoxyxylostasin, 1N- [L - (-) - γ-amino-α-hydroxybutyryl] paroinomycline I, 1-N- [L - (-) - 7 -amino-o-hydroxybutyryl] -3'-deoxyparomomycin I, 1N- [L - (-) - 7-amine-e-hydroxybutyryl] -3 \ 4'-dideoxyparomomycin I, 1-N- [L - (- ) -7-amino-o-hydroxybutyryl] paromomycin II, 1-N- [L - (-) - 7-amino-o-hydroxybutyryl] -3'-deoxyparomomycin η 11,25 lN- [L - (-) - γ-amtno-α-hydroxybutyryl] -3 ', 4'-dideoxyparomomycin II, 1-N- [L - (-) - 7-amtno-o-hydroxybutyryl] aminoglycoside 2230-C, 1-N- [L- ( -) - 7-Amino-o-hydroxybutyryl] -3'-deoxy-aminoglycoside 2230-C, 1N- [L - (-) - 7-Amino-o-hydroxybutyryl] 11vidomycin A and IN- [1- (-) -7-amino-o-hydroxybutyryl] 11-yldomycin B.

30

Reaction of each of the above carbobenzyloxy protected aminoglycoside starting materials in the same manner with L - (-) - 0-benzyloxy-carbonylamino-α-hydroxypropionic acid N-hydroxy-5-norbornene-2,3-dicarbox-Imide ester instead for the L - (-) - γ-benzyloxycarbonylamino-o-hydroxybutyric acid-N-hydroxy-5-norbornene-2,3-dicarboxylmethyl ester produces the corresponding 1N- [1 - (-) - 0-amino-e-hydroxypropylonyl] amt noglycos1der.

Reaction of each of the above carbobenzyloxy protected aminoglycoside starting materials in the same manner with L - (-) - i-benzyloxycarbonylamino-o-hydroxyvaleric acid-N-hydroxy-5-norbornene-2,3-BNS DOCID: <DK 1725438 »46 DK PR 172543 B1 dicarboxyl moiety ester in place of the L - (-) - T-benzyloxycarbonylamino-o-hydroxy acid acid N-hydroxy-5-norbornene-2,3-dicarboxine nudester produces the corresponding 1N- [L - (-) - 6 -araino-ol-hydroxyvaleryl] amino-glycosides.

5

Example 30

The general procedure of Example 1 is repeated except that 6'-N-carbobenzyloxycanamycline A used therein is replaced by an equimolar amount of 10 2 ', 3,6'-tr1- (N-carbobenzyloxy) -3', respectively. 4'-dideoxyri bostamycin, 2 ', 3,6'-tri- (N-carbobenzyloxy) -3', 4'-dideoxyneoniycin B, 2 ', 3,6'-tri- (M-carbobenzyloxy) -3', 4 # -dideoxyneomycln C and 2 ', 3,6'-tri- (N-carbobenzyloxy) -3', 4'-di-deoxylostasin, 15 thereby producing 1-N- [L - (- Jy-amino-a) -hydroxybutyryl] -3 ', 4-dideoxyribostamycin, 1-N- [L - (-) -γ-aalno-α-hydroxybutyryl] -3', 4'-dideoxyneomyclη B, 20 lN- [L - (-) - γ-amino-α-hydroxybutyryl] -3 ', 4'-dideoxyneomycline C and 1N- [L - (-) - 7-amino-e-hydroxybutyryl] -3', 4'-dideoxyxyostostine.

Reaction of each of the above 2 ', 3,6'-tri- (N-carbobenzyl oxy) protected aminoglycoside starting materials in the same manner with 25 L - (-) - ^ - benzyloxycarbonylamino-α-hydroxyproplonic acid N-hydroxy The 5-norbornene-2,3-dicarboxylmidester in place of the 1 - (-) - γ-benzyloxy-carbonyl-amino-α-hydroxybutyric acid-N-hydroxy-5-norbornene-2,3-dicarboxylin ester gives the corresponding 1N- [ L - (-) - 0-am1no-ft-hydroxyprop1onyl] am1no-glycosides.

Reaction of each of the above 2 ', 3,6'-tri- (N-carbo-benzyloxy) protected aminoglycoside starting materials in the same manner with L - (-) - i-benzyloxycarbonylamino-a-hydroxyvaleric acid-N The hydroxy-5-norbornene-2,3-dicarboxylic acid ester instead of the L - (-) - γ-benzyloxy-carbonylamino-e-hydroxybutyric acid N-hydroxy-5-norbornene-2,3-dicarboxylic acid ester produces the corresponding lN- (L - (-) - £ -amino-e-hydroxy-valeryl] am1noglycos1der.

nwcnnnri 47 DK PR 172543 B1

Example 31

Preparation of 1N-fL-Mv-amino-tt-hydroxybutyryl] -1 - dexoxy-e'-N-methylcanamycin A (BB-K3111 by acylation of polysilylated 6'-N-benzyl-3-M-carbobenzyloxy-4'-deoxy -6'N-Nitylcyanamycin A 5 A. 6'-N-carbobenzyloxy-4'-deoxyquanamycin A and 3,6'-di-N-carbobenzyloxy-4f-deoxycanamycin A To a stirred solution of 10.4 g (22, 2 mmol) of 4'-deoxycanamycline A and 27.5 g (111 moles) of Ni (OAc) 2-4 H2 O in 450 ml of ONSO were added 7.0 g (22.4 10 mmol) of N-hydroxy-5-norbornene-2 The 3-dicarboximide ester of benzyloxy-formic acid (Cbz-ONB) was stirred at about 10 ° C and stirred overnight at room temperature. The reaction mixture was concentrated in 1 vacuum to give a bl4 oily residue which was chromatographed over "CG-50" resin (ΝΗ4 *, 500 ml), eluting with dilute ammonia water, the 15 ninhydrin positive fractions collected from the waste (due to a leakage of 3,6'di-N-Cbz derlvate) and the eluate with 0.1 N ammonia was combined and evaporated in vacuo. The residue was chromatographed over "Diaion HR-10" resin (400 ml) using aqueous methanol as eluent to separate the mono- and di-Cbz derivatives. The 6'-N-Cbz-20 derivative eluted with 30% aqueous methanol was crystallized from HgO-ethanol to give 4.73 g (35%) of colorless crystals. Mp. 232-233®C. IR (KBr): 1700, 1540, 1275, 1135, 1075, 1040, 770, 750, 695 cm -1.

Analysis calculated for C CHH ^ NN₂O ^EtOH. ^O: C, 51.13; H, 7.51; N, 8.52.

Found: C, 51.03; H, 7.31; N, 8.42.

The 3,6'-di-N-Cbz derivative, eluted with 90% aqueous methanol, was crystallized from HgO-methanol to give 3.08 g (18.4%) of nile, m.p.

220-221 ° C, IR (KBr): 1690, 1545, 1260, 1135, 1075, 1045, 780, 750, 700, 30 cm -1.

Analysis calculated for C34 H48 N4 ° 14 H2 O: C, 54.10; H, 6.68; N, 7.42.

Found: C, 54.10; H, 6.69; N, 7.07.

B. 1.S.a'-tri-N-acetyl-B'-N-carbobenzylPXV-1'-deoxy-kananycin A

To a stirred suspension of 4.70 g (7.8 mmol) of 3,6'-di-NCbz-4'-deoxycanamycin A in 240 ml of methanol was added 29 ml (307 mmol) of acetic anhydride. The mixture was stirred overnight at room temperature and evaporated in vacuo to give 5.68 g (100%) of the title BNS DOCID: <OK 172S43B> 48 DK PR 172543 B1, tr1-N-acetyl derivative, m.p. 290-291 ° C (decomp.), IR (KBr): 1700, 1650, 1550, 1380, 1260, 1140, 1080, 1035, 745, 695 cm -1.

Analysis calculated for γ · Η, O: C, 51.47; H, 6.75; N. 7.50.

Found: C, 51.66; H, 7.13; N, 7.06.

C. 1.3.3 "-Tri-N-acetrl-4'-deoxycanamycin A

A solution of 5.5 g (7.6 mmol) of 1,3,3 "tri-N-acetyl-6'-N-carbobenzyloxy-4'-deoxycanamycline A in 80 ml of 50% aqueous ethanol was hydrogenated in 18 hours with 1 g 10% Pd-C at atmospheric pressure and room temperature The catalyst was removed by filtration and the filtrate was evaporated under reduced pressure to give 4.49 g (100%) of the title compound. by column chromatography on "Amberlite IRA-410" resin (OH ') and crystallization from 15 H 2 O methanol, mp 280-283 ° C, IR (KBr): 1650, 1550, 1380, 1135, 1080, 1035 cm -1.

Analysis calculated for C C HH ^NfO ^ -NeOH.fcHgO: C, 47.24; H, 7.45; N, 8.81.

Found: C, 47.61; H, 7.45; N, 8.58.

20

0. 1.3.3 "-Tri-N-acetyl-6'-N-benzyl-4 * -deoxycanamycin A

A solution of 4.39 g (7.39 mole) of 1,3,3 "tri-N-acetyl-4'-deoxycanamycline A and 5 ml of benzaldehyde in 60 ml of aqueous methanol was heated to 60 ° C for 30 minutes. was cooled, treated with 4.0 g (106 mol) of NaBH 3, stirred for 2 days at room temperature and evaporated in vacuo. The oily residue was chromatographed over "HP-10" resin (150 ml) using aqueous methanol as eluent. portion to give 3.86 g (74%) of the title compound and 1.70 g of moist starting material which was recycled in the same process to give an additional 1.32 g (26%) of the 1 An analytical sample was prepared by chromatography on "HP-10" resin and crystallization from HgO-ethanol, m.p. > 300 ° C, IR (KBr): 3280, 1640, 1555, 1380, 1135, 1080, 1040, 750, 700 cm -1.

Analysis calculated for; 35 C, 54.38; H, 7.07; N, 8.18.

Found: C, 54.01; H. 7.16; N, 7.87.

E. 1, 3,3 "-tr1-N-acetyl-6, -N-benzyl-4> -deoxy-6, -N-methylphenylamycin A To a stirred solution of 3.86 g (5.6 mmol) 1 , 3,3 "-tri-N-acetyl-aNsnoon · <· ηκ i7? Wsr» 49 DK PR 172543 B1 6'-N-benzyl-4'-deoxycanamycin A and 5.6 ml 37¾ aqueous HCHO in 85 ml 95X methanol was added 850 mg (13.5 mmol) of NaBH 3 CN. The mixture was stirred for 4 hours at room temperature and evaporated to dryness. The residue was chromatographed over "HP-10" resin using aqueous methanol as eluent to give 3.88 g (100¾) of the title compound. An analytical sample was obtained by crystallization from H 2 O-ethanol, m.p. 295-297 ° C (decomp.). IR (KBr): 3280, 1645, 1500, 1375, 1145, 1075, 1040, 740, 700 cm -1. NMR (DgO): S in ppm from DSS, 2.27 (3H, s, N-CH 3), 7.34 (5H, s, Ar-H).

Analysis calculated for ε ^ Η ^ Ν ^ Ο ^. ^ Ο: C, 54.30; H, 7.26; N, 7.92.

Found: C, 54.30; H, 7.33; N, 7.64.

F. S'-N-benzyl-A'-deoxy-S'-N-methylcanamycin A (BB-K312) A mixture of 5.18 g (7.42 mmol) of 1,3,3 "tri-N -acetyl-6'-N-benzyl-4'-deoxy-6'-N-methylkanamycin A and 15 g of NaOH in 80 mL of valΗ 0 to reflux were added overnight. The mixture was cooled, neutralized with wife, HCl and filtered to The filtrate was introduced on top of a column of "CG-50" resin (NH 2 +, 370 ml). After washing with 1.5 20 liters of water, the column was eluted successively with 2 liters of 0.05 n NH 2 OH and 2 liters of 0.1 n NH 4 OH.

The eluate, which showed positive ninhydrin reaction, was evaporated to give 3.23 g (76¾) of the title compound.

6. 6'-N-benzyl-3-N-benzyloxycarbonyl-4 * -deoxy-6'-N-methyl-kanamvein A

To a stirred solution of 2.94 g (5.14 mmol) of 6'-N-benzyl-4'deoxy-6'-N-methylkanamycin A (BB-K312) and 6.37 g (25.7 mmol) of Ni (OAc) 2 * 4H 2 O in 100 ml of ONSO was added 1.62 g (5.20 mmol) of Cbz-ONB and stirring was continued at room temperature. After 2 days, 72 ml of 0.2 m aqueous EDTA solution and 30 ml of wife were added. NH.OH to the reaction mixture. The blue solution obtained was introduced on top of a column of "HP-10" resin (150 ml). After washing with 200 ml of 7 n NH 2 OH followed by 300 ml of H 2 O, the column was eluted with 400 ml of 80X aqueous methanol to give 2.65 g (73¾) of the title compound. An analytical sample was prepared by crystallization from methanol, m.p. 216-217 ° C. IR (KBr): 3340, 1690, 1540, 1290, 1135, 1045, 745, 700 cm -1.

Analysis calculated for C, 57.06; H, 7.18; N, 7.83.

Found: C, 57.29; H, 7.22; N, 7.58.

BNS DOCID: <DK 172543B> 50 DK PR 172543 B1 H. 1-H-fL- (Q-'Y-amino-tt-hydroxybutyrryl-4> -deoxy) -6, -N-methylcananecin A_ (BB-K3111

A suspension of 2.49 g (3.53 mmol) of 6'-N-benzyl-3-N-benzyl-oxycarbonyl-4'-deoxy-6'-N-methylcanamycin A and S for back dropping oats overnight. The one achieved clear resolution wife one third to dryness. To a stirred solution of the oily residue in 50 ml of diethyl ketone was added 1.23 g (3.51 nmol) of SAE. The mixture was stirred overnight at room temperature and evaporated in vacuo to give an oily residue, which was treated with HgO-ethanol, adjusted to pH 10. 3 with 1 n HCl and allowed to stand for 30 minutes at room temperature. This solution containing the acylated product (a small sample isolated from the solution showed a carbonyl absorption at 1655 cm -1 due to an amide, was hydrogenated overnight with 1 g of 10% Pd-C at atmospheric pressure and room temperature. was removed by filtration, 15 and the filtrate was evaporated in vacuo. The residue was dissolved in a small amount of water and the solution was introduced into the top of a column of "CG-50" resin (NH 4, 160 ml). the column successively down 250 ml of 0.1 η NH 2 OH, 1 liter 0.2 n NH 2 OH and finally 1 liter 0.5 n NH 2 OH. The eluate was collected in 20-ml fractions. Fractions 89-100, which 20 showed positive ninhydrin reaction, evaporated in vacuo to give an oily residue, the residue was solidified in H 2 O-ethanol to give 1.25 g of crystalline product and 144 mg of a second portion, totaling 399 g (68%) of the title product, mp 182-184 ° C, [a) lZl 93 »(c 1, H2 O).

Analysis calculated for C 23 H 45 N 5 ° 12 * H * H 2 O: C, 46.36; H, 8.25; N, 10.81.

Found: C, 46.37; H, 8.40; N, 10.38.

This product was chromatographed over "CG-50" resin (NH 4 +, 100 30 ml) using carbonate-free aqueous ammonia as eluant and crystallized from H 2 O-methanol-isopropanol to give 927 mg of the title compound as colorless needles, m.p. . 193-194 »C, [e] J2 + 101« (c 1, H2 O). NMR (020): S in ppm from DSS, 1,37 (2H, q, J-12,2-Hax & 4-Hax), 1.86 (4H, m, 2-Heq, 4'-Heq & -CH2), 2.29 (3H, s, N-CH2), 4.14 (1H, dd, J-8 & 4.5, a-CH), 5.00 (1H, d, J- 3.5, 1'-H), 5.24 (1H, d, J '3.5, rH). IR (KBr): 1640, 1540, 1135, 1080, 1040, 955 cm -1.

Analysis calculated for Ν, -0 2H, 0: C, Μ.μΓη; M7, ». * 11.30

Found: C, 44.85; H, 7.90; N, 11.85.

^ Mcnnrin 51 DK PR 172543 B1

Example 32

Preparation of 1-N-fL-M-T-amino-tt-hydroxybutyrene-4'-deoxycanamycline A (BB-K160) by acylation of polysilylated 3.6 * -d1-M'cerbobenzylphxy.-AL ·

S deoxycarbonamide A

A suspension of 2.27 g (3.08 mmol) of 3,6'-di-N-carbobenzyloxy-4'-deoxycanamydn A and 5 ml of HHDS in 35 ml of dry CH 2 CN is refluxed for 2 days. The resulting clear solution was concentrated in vacuo. The oily residue was dissolved in 40 ml of dry diethyl ketone. To the solution was added 1.08 g (3.08 mmol) of SAE with stirring. The mixture was stirred overnight at room temperature and evaporated to dryness. The residue was treated with ethanol-H 2 O and adjusted to pH 3 with 1 n HCl. After standing for 30 minutes, the acidic solution was hydrogenated for 3 days with 1.2 g of 10% Pd-C at atmospheric pressure and room temperature. The catalyst was removed by filtration and the filtrate was evaporated in vacuo. The residue was dissolved in a small amount of water and the solution was introduced into the top of a column of "CG-50" resin (NH₂ +, 130 ml). After washing with 500 ml of H 2 O, the column was eluted successively with 1 liter of 0.2 n NH 2 OH and 1 liter 0.5 n NH 2 OH. The eluate was collected in 20-ml fractions. Fractions 79 - 95, which showed a positive 20 ninhydrin sample, were combined and evaporated in vacuo to give 1 g (56, IX) of the title compound as an amorphous powder. The product was crystallized by the same procedure as that used in Step H of Example 31 for BB-K311, m.p. 179-180 ° C, [α] 21 ° + 99 ° (c 1, H 2 O). IR (KBr): 1640, 1535, 1120, 1065, 1030, 945 cm. NMR (D20): δ in ppm, 1.37 (2H, q, J-12.2-4.4'-Hax), 1.86 (4H, m, 2- & 4'-Heq & O-CH2) , 4.13 (1H, dd, J-8 & 4,5,0-CH), 5.02 (1H, d, J-3, Γ-Η), 5.28 (1H, d, J »3 , 5, l'-H).

Analysis calculated for C 22 H 43 N 5 ° 12 * MeOH * 2H 2 O: C, 43.47; H, 7.95; N, 11.27.

Found: C, 43.69; H, 7.92; N, 10.91.

BNS DOCID: <DK 172543B>

Claims (56)

A process for the preparation of 1-N - [[α] -amino-e-hydroxy-alkanoyl] aminoglycosides of formula (I) NH NH, R 5 in (i) K RO C = 0 2. A process according to claim 1, characterized in that the acylated derivative of the acid of formula (XIII) is an active ester or a mixed acid anhydride. 3. A process according to claim 1 or 2, characterized in that the amino blocking group p in the acylating derivative of the acid of formula (XIII) is selected from groups of formulas 30 R20 O 0 CH 3 0 0 / ΓΛ ii in ii ii (f y -ai2oc-, CH-jcoc-, * 2xcc-, 35 "3 RNSOOCin- <DK 172543B 9 55 DK PR 172543 B1, Q ~ - Of- · iQ) o 4. A process according to any one of claims 1-3, characterized in that the acylating derivative of the acid of formula (XIII) is its active ester with N-hydroxysuccin1m1d, N-hydroxy-5-norbornene-2,3- dicarboximide or N-hydroxyphthalimide. 5 JR represents H or OH, and 4 20. represents H, OH or a pentofuranosylation of the formula: Μ0 <: | * 2 Ov, <0 <iV0 \ I ^ 7 ·· 'yj OH KO 0, i (IX) ( X) wherein R represents H or a hexopyranosyl ring of the formula ch2nh2 ΛΜ__ο ,, ΛΤΛ · “· (χΐ) (XU) ; on the assumption that when R is different from H, one of the groups R4 and R5 represents H and the other OH; and under the premise that 3.a 5 when R represents H, R H and R represent a pentofuranosyl ring of formula (IX) or (X); with silylating agents of the formulas 10 «v r ϊ ----- NH K14 NH R15 V \ l / R14 16 I ^ Sl. / and Ri6-Si-Z 15 r14 \ n - Si '\ H \ R \ n14 R m L / (XV) (XVI) 20 wherein R15, R16 and R17 are selected from hydrogen, halogen, (lower) alkyl , (lower) alkoxy, halogen (lower) alkyl and phenyl, wherein the 25 'moiety of one of the groups R * ®, R ^ and R ^ 7 is different from halogen or hydrogen; R 14 represents (lower) alkyl, 'is an integer of 1 to 2 and Z is selected from halogen and H0] CH3 HO I (V) (VI) 5 R 9 Q ~ · O-f— · 15 o s X, C-CII, -0-C- and. # 5 R ”2 / NH2 Γ HO Fri. 5. A process according to any one of claims 1-3, characterized in that the acylating derivative of the acid of formula (XIII) is its mixed anhydride with pivalic acid, benzoic acid, isobutylcarbonic acid or benzylcarbonic acid. Wherein R 2, R 3, R 4 and R 5 have the meaning defined above, and zone 10 optionally contains from 1 to 3 amino-blocking groups which are different from sllyl on amino groups other than the Cl amino group, in an organic solvent, which may contain up to approx. 40% water, with an acylating derivative of an acid of formula (XIII) 5 52 DK PR 172543 B1 ΡΑΙζΝΤΚΜγ 6. A process according to claim 4 or 5, characterized in that the amino blocking group on the acylating derivative of the acid of formula (XIII) is carbobenzyloxy, tr1 fluoroacetyl or 1-butoxycarbonyl. 7. A process according to claim 6, characterized in that the polysilylated aminoglycoside contains from 1 to 3 aminobloxy groups in the form of carbobenzyloxy or trifluoroacetyl on amino groups other than the C-1 amino group. Method according to any one of claims 1-7, KENOE-BNSDOCIDkDK 172543B> 56 DK PR 172543 B1 TE6NET in that the sHyl groups are trimethylsilyl groups. A process according to claim 1 for the preparation of 1N- [L - (-) - u-amino-o-hydroxyalkanoyl) aminoglycosides down the formula 5 R6 / K nch-r26 R1 ° W - ___ r9 ~ 8s rI3 M / / c = o * / / HO-CH t - (-) / (C "2, n" 24/1 R 25 / CH- .s "iA-I'-ox / r22H" A --- --- 7n \ / 110 / o 20 wherein n is an integer of from 0 to 4, R6 represents H or CH3, R® represents OH or NH2, R9 represents H or OH, R10 represents H or OH, R22 represents H or CH3, R23 represents OH or CH3, R2 * represents H or OH, R25 represents H or CH2 OH, and R26 represents OH, NH2 or NHCH, provided that when R22 represents H, R2® represents CH.OH And * when R represents CH-, RH, and provided that when 23 * 2a 23 2A R represents OH, RH represents, and when R represents CH 2, R represents OH, or pharmaceutically acceptable salts thereof, by using as a polysllylated aminoglycoside a 30 polysllylated aminoglycoside prepared from an aminoglycoside having to the formula 35 ^ Msnorin 57 DK PR 172543 B1 R6 „lo'Kj" -1'26 s //, l0 ^ y ^ Vwl2 ° / / Γ »" / 10 "Y - ° \ / - ~ 7χ \ / 1. wherein R6, R8, R9, R10, R22, R23, R24, R25 and R26 are as defined above and optionally contain from 1 to 3 amino-blocking groups which are different from silyl in amino groups other than The Cl amino group, and using as an acylating derivative an acylating derivative of an acid of formula (XIII) L - (-) B-HN-CH 2 - (CH 2) n-CH-COOH (XIII) OH 25 wherein B and n has the meaning defined above. Wherein R9 represents H or OH, R * ® represents H or OH, R2® represents OH, NH2 or MHCH2, R29 represents H or OH, and R30 represents H, OH or OR31, wherein R31 represents a hexopyranosyl ring of formula Z0 33 CH2N1I2I! »Η -ΜHN] 32 wherein R represents H or α-D-mannopyranosyl, and one of the groups 10 HO j 2 ° H j ψ h3chn / / H0V \ ____ H3C ° / \ XX_ | W or H2N νΛ ch3 “Ί (VII) (VIII) Λ 1 wherein R R represents H or CH,; HOClf, n / \ ts ° \ / 2jPj ---- {J, 30 R OH 15 wherein R9, R10, R26, R29 and R30 are as defined above and optionally contain from 1 to 3 amino blocking groups which are 20 different from sllyl on amino groups other than the Cl amino group, and that as an acylating derivative an acylating derivative of an acid of formula (XIII) 1- (-) B-HN-CH 2 - (CH 2) n-CH-C XIII) Process according to claim 9, characterized in that the acylating derivative of the acid of formula (XIII) is an active ester or a mixed acid anhydride. 10 O 11 «11 X3C-CH2-0-C- and. wherein R 2 O and R 21 are the same or different and each is 1 represents H, F, Cl, Br, NO 2, OH, (lower) alkyl or (lower) alkoxy, X represents Cl, Br, F or I and Y represent H, Cl, Br, F or I. 10 in HO-CH (iH2Jn CH, I in nh2 15 wherein n is an integer of from 0 to 4; R2 represents a hexopyranosyl ring of the formula f R6 R6 A process according to any one of claims 9 and 10, characterized in that the amino-blocking group of the acylating derivative of the acid of formula (XIII) is selected from groups of the formulas A process according to any one of claims 9-11, KEN · DETE6NET in that the acylating derivative of the acid of formula (XIII) is its active ester with N-hydroxysuccin1mfd, N-hydroxy-5-norbornene-2,3 - dicarboxylmide or N-hydroxyphthalimide. A process according to any one of claims 9-11, characterized in that the acylating derivative of the acid of formula (XIII) is its mixed anhydride with plvalic acid, benzoic acid, isobutylcarbonic acid 30 or benzylcarbonic acid. A process according to claim 12 or 13, characterized in that the amino-blocking group on the acylating derivative of the acid of formula (XIII) is carbobenzyloxy, trifluoroacetyl or -t-butoxycarbonyl. 35 15 NET knows that the silyl groups are trimethylsilyl groups. HO I / O wherein R 22 represents H or CH 2 and R 28 represents H or CH 2. 20 15. A process according to claim 14, characterized in that the polysilylated aminoglycoside contains from 1 to 3 amino blocking groups in the form of carbobenzyloxy or trifluoroacetyl in amino groups other than the ΟΙamino group. . »Nsoorm · 59 DK PR 172543 B1 B-HN-CH 2 - (CH 2) n-CH-COOH (XIII) OH wherein B represents an amino-blocking group and n has the meaning defined above; and then removing all blocking groups. Process according to any one of claims 9-15, characterized in that the silyl groups are trimethylsilyl groups. A process according to claim 1 for the preparation of 1-N- [L - (-) - u -amino-hydroxyalkanoyl) aminoglycosides of the formula r27-chnh-r28 NH, 10 / vi2 h2n »0 / \ HO /. / C = O / / HO-Cl 1 L - (-) 15 - - Q / <CH 2) n HjCHN - ^ - / T 2 HO 1 / NH, O 2 20 or a pharmaceutically acceptable salt thereof, wherein n represents a represents numbers from 0 to ♦, R 30 -Λ H2N \ ----- 1W Oil / X == \ / 35 h3chn '^ 7n \ / «ο y BNSDOCID; <DK 172543B> 60 DK PR 172543 B1 P7 pe wherein R and R have the meaning defined above and which contain 2 or 3 amino blocking groups, are different from silyl on other amino groups than the C1 amino group and that acylating derivative is used an acylating derivative of an acid of formula (XIII) L - (-) B-HN-CH2 - (CH2) n-CH-C00H (XIII) OH 10 wherein B and n have the meaning defined above. 18. A process according to claim 17, characterized in that the acylating derivative of the acid of formula (XIII) is an active ester or a mixed acid anhydride. 15 A process according to claim 17 or 18, characterized in that the amino blocking group on the acylating derivative of the acid of formula (XIII) is selected from groups of formulas _20 20 *, -v O CK, OO ii in 3 ii ii (f y ---- ch2oc-, ch3-coc-, y2xcc-, r21 ^ K = z / ch3 o Q-- 'OT -..... ”oo 35» X, C-CH, -0-C- and VN. II OCID: <DK 177643B. 61 DK PR 172543 B1 20 21 wherein R and R are the same or different and each represents H, F, Cl, Br, NO 2, OH, (lower) alkyl or (lower) alkoxy, X represents Cl, Br, F or I, and Y represents H, Cl, Br, F or I. Wherein R and R are the same or different and each ice year represents H, F, Cl, Br, NO2, OH, (lower) alkyl or (lower) alkoxy, X represents Cl, Br, F or I and Y represents H, Cl, Br, F or I. 25 A process according to any one of claims 17-19, characterized in that the acylating derivative of the acid of formula (XIII) is its active ester with N-hydroxysuccinimide, N-hydroxy-5-norbornene-2,3-dicarboximide or N-hydroxyphthalimide. R10 is V \ CHNHR7 J.HOM CHNHR7 1ΖΓ ° ν r1 ° x \ n ^ - - o - TV 25 (II) (III) (IV) wherein R® represents H or CH 2, R 7 represents H or CH 2, R® represents OH or HH ^, R® represents H or OH, and R * ® represents H or OH; R3 represents H or a hexopyranosylation of the formula Γ,, n ^ —'- Λ 35 2 7 Λ '---' H0 1 CH3 »O I (V) cVI) NS NSDOCID <OK 172543B> 53 DK PR 172543 B1. S, uo ^ · «« (twist) (wine) 2 In 10 wherein R11 represents H or CH3; R 5 represents H or OH and R * represents H, OH or a pentofuranosyl ring down the formula "hccl> °" -4 v_7 -> - / OH R 1 O 2 OH (IX), xl 20 wherein R1Z represents H or a hexopyranosyl ring of formula CH2nh2. .... "- VK Αη *, jA (XI) (XII) 30 wherein R * 3 represents H or oD-mannopyranosyl; 3 under the assumption that nir R is different from H, ic represents one of the groups R and RH and the other OH; and under the precedent that n R represents H, RH and R represent a pentofuranosyl ring of formula (IX) or (X), or pharmaceutically acceptable acid addition salts thereof; a polysyllylated aminoglycoside prepared from an aminoglycoside of formula (XIV) BNS DOCID is reacted: <DK 172543B »54 DK PR 172543 B1 yw nh2 -> (XIV) A process according to any one of claims 17-19, characterized in that the acylating derivative of the acid of formula (XIII) is its mixed anhydride with plvalic acid, benzoic acid, isobutylcarboxylic acid or benzyl carboxylic acid. A process according to claim 20 or 21, characterized in that the amino blocking group on the acylating derivative of the acid of formula (XIII) is carbobenzyloxy, trifluoroacetyl or -butoxycarbonyl. A process according to claim 22, characterized in that the polysilylated aminoglycoside contains 2 or 3 amino blocking groups in the form of carbobenzyloxy or trifluoroacetyl on amino groups other than the C-1 amino group. The method of any of claims 17-23, Cl, Br, NO 2, OH, (lower) alkyl or (lower) alkoxy, X represents Cl, Br, F or I, and Y represents H, Cl, Br, F or I, in amino groups other than the C-1 amino group. In OH wherein B and n have the meaning defined above. The process of claim 1 for the preparation of 1N- [L - (-) - w -amino-α-hydroxyalkanoylamino] glycosides of formula 30 BNS DOCID: <DK 172S43B »62 DK PR 172543 B1 I / HO-CH L - (-) 10 iii, x x γ ί "2 RII nh, R30 Oli 15 each n represents an integer pi from 0 to 4, R9 represents H or OH, R * ° represents H or OH, R26 represents OH, NH2 or NHCH2, R29 represents H 20 or OH, and R30 represents H, OH or OR31, wherein R31 represents a hexopyranosyl ring down the formula "33 η2ν I 32 32 wherein R represents H or α-D mannopyranosyl, and one of the groups 33 34 R and R represents H and the other CHgNHg; assuming that nir R29 represents H, R30 represents OH or OR31 and that nir R29 represents OH, R30 represents H, or pharmaceutically acceptable acid addition salts thereof; CHARACTERISTICS: By using as a polysyllylated aminoglycoside, a polysilylated aminoglycoside prepared from an aminoglycoside of the formula is used. ^ SDOCIO <Γ »Κ 172S43B> 63 DK PR 172543 B1 / K CH2-R26 ^ ± -r \ THE CHARACTERISTICS OF THE SULYL GROUPS are trimethylsilyl groups. Process according to claim 25, characterized in that the acylated de derivative of the acid of formula (XIII) is an active ester or a mixed acid anhydride. A process according to claim 25 or 26, characterized in that the 35 amino blocking group on the acylating derivative of the acid of formula (XIII) is selected from the groups of formulas BNS DOCID: <OK 172S43B> 64 DK PR 172543 B1 R20 _ O CH, O 0 y ^ S - CIljOC-, CHj-COC-. * 2xcc-. 213 = / CH3 28. A process according to claim 27, characterized in that the acylating derivative of the acid of formula (XIII) is its active ester with N-hydroxysuccinimide, N-hydroxy-5-norbornene-2,3-dicarboxylmid or N-hydroxyphthalimide. 30 29. A process according to claim 27, characterized in that the acylating derivative of the acid of formula (XIII) is its mixed anhydride with pallic acid, benzoic acid, isobutylcarbonic acid or benzylcarbonic acid. 30 NET knows that the syllyl groups are trimethylsilyl groups. R33 and R3 * denote H and the other CHgNHgi, provided that when R29 represents H, R3 represents OH or OR3 * and that when R29 represents OH, R3 represents H. Wherein R represents hydrogen or (lower) alkyl and R represents hydrogen, (lower) alkyl or R 15 -15- BNSDOOID <DK 172543B> 68 DK PR 172543 B1 wherein R 1, R 16 and R ^ 7 has the meaning defined above, and is also desired by the introduction of 1-3 amino-blocking groups of formulas R20 __ O CH3 OO 5 -c "2 ° ^" 'CH3-C-O-C-, 2XCC-. u21 3 o Q-- o-j · - · o o 20 ίί X3C-CU2-0-C- 09 IH ^ ίί o 20 21 wherein R and R are the same or different and each 1s * r represents H, F, A process according to any one of claims 25-29, characterized in that the amino blocking group on the acylating derivative of the acid of formula (XIII) is carbobenzyloxy, trifluoroacetyl or t-butoxycarbonyl. R ^ SDOCID: <DK 17P543B> 65 DK PR 172543 B1 31. A process according to claim 30, characterized in that the polysilylated aminoglycoside contains from 1 to 3 amino blocking groups in the form of carbobenzyloxy or trifluoroacetyl on amino groups other than the rupp amino group. 5 32. A process according to claims 25-31, characterized in that the silyl groups are trimethylsilyl groups. 33. Polysilylated aminoglycoside having 2 * 10 silyl groups for use * 10 as a starting material in the practice of the process of claim 1 for the preparation of 1N- [w-afflino-α-hydroxyalkanoyl] aminoglycosides of the formula (I), wherein it is prepared by reacting an aminoglycoside of formula (XIV) wherein R represents a hexopyranosyl ring of formula: r10W "K", R7 ^ Ihoh f- »7.. ... * (II) (III) (IV) wherein R6 represents H or CH3, R7 represents H or CH3, R8 represents OH or NHZ, R9 represents H or OH, and R10 represents H or OH; R3 represents H or a hexopyranosyl ring of the formula: BNSDOC1D: <OK 172543B> 66 DK PR 172543 B1 H2 ^ 7n \, UN * 34. Polysllylated aminoglycoside According to claim 33, characterized in that it contains an average number of sllyl groups per cell. molecule pi from 3 to B. The polysyllylated aminoglycoside according to claim 33 or 34, characterized in that the silyl groups are trimethylsilyl groups. 35 35 R20 —λ,? CH3 0 0 C \ -ch2oc-, ch -c-o-c-. Y xcc-, r21 ^ A = / CH3 BNS DOCID: <DK 172543B> 58 DK PR 172543 B1 G. Q ~ Of- '> »2 o 10 o li f' X3C-CH2-0-C- and -I jl VGN .. ^ II 15 0 20 21 wherein R and R are the same or different and each ismr represents H, F, Cl, Br, NOg, OH, (lower) alkyl or (lower) alkoxy, X represents Cl, Br, 20. or I, and Y represents H, Cl, Br, F or I. The polysyllylated aminoglycoside according to any one of claims 33-35, KEM) ETE6NET, in that the amino blocking groups are carbo-benzyloxy or trifluoroacetyl groups. HNsnorin, ηκ ι7 ·> μιβ »69 DK PR 172543 B1 37. Polysyllylated aminoglycoside According to claim 33, characterized in that it is prepared by reaction of an aminoglycoside of the formula 5 "* 10 1 / / lv 'o / / r2-1 ^ / IS -" ° \ / "" .. Wherein R6 represents H or CH3, R® represents OH or NHg, R9 represents H or OH, R10 represents H or OH, R22 represents H or CH2, R23 represents OH or CH3, R2 * represents H or OH, R2® represents H or CH2 OH, and R26 represents OH, NH4 or NHCH2 *. provided that when R 22 represents H, R 25 represents CH 2 OH and when R 22 represents CH 3, R 2 represents H; and provided that when R23 represents OH, R2 represents <H, and when R23 represents CH3, R2 represents OH. 38. Polysllylated aminoglycoside According to claim 37, characterized in that it contains an average number of sllyl groups per molecule of from 3 to 8. 39. Polysyllylated aminoglycoside According to claim 37 or 38, characterized in that the silyl groups are trimethylsilyl groups. 35 40. Polysilylated aminoglycoside According to any one of claims 37-39, characterized in that the amino blocking groups are carbo-benzyloxy or trifluoroacetyl groups. BNSOOCID: <DK 172543B> 70 DK PR 172543 B1 41. Polysylated aminoglycoside According to Claim 33, characterized in that it is prepared by reacting an aminoglycoside of formula 5 57 28 R 1 -CHNH-R H, N L / \ 2 _ \ ^^ 7N \ / Nii 42. Polysilylated aminoglycoside According to claim 41, characterized in that it contains an average number of sllyl groups per molecule pi from 3 to 8. 43. Polysilylated aminoglycoside According to claim 41 or 42, KENDETEG NET knows that the silyl groups are trimethylslyl groups. 44. Polysilylated aminoglycoside According to any one of claims 41 - 43, characterized in that the amino blocking groups are carbobenzyloxy or trifluoroacetyl groups. 45. Polysilylated aminoglycoside according to claim 33, characterized in that it is prepared by reacting an aminoglycoside of the formula 35> .gSDOrin · -OK 172543B> 71 DK PR 172543 B1 / vr 46. A polysyllylated aminoglycoside according to claim 45, characterized in that it contains an average number of silyl groups per cell. molecule of from 3 to 8. 47. Polysilylated aminoglycoside according to claim 45 or 46, CHARACTERISTIC BNS DOCID: <DK 172543Β »72 DK PR 172543 B1 NET in that the silyl groups are trimethylsilyl groups. 48. Polysilylated aminoglycoside according to any one of claims 45-47, characterized in that the amino blocking groups are carbobenzyloxy or trifluoroacetyl groups. The polysilylated aminoglycoside of claim 33, characterized in that it is polysilylated by the blending of gentamicin Bj. 50. A polysilylated aminoglycoside according to claim 49, characterized in that it contains an average number of silyl groups per cell. molecule of from 3 to 7. The polysilylated aminoglycoside of claim 49 or 50, 52. A polysilylated aminoglycoside according to any one of claims 49-51, characterized in that the amino blocking groups are carbo-benzyloxy or trifluoroacetyl groups. 20 53. The polysilylated aminoglycoside of claim 33, characterized in that it is polysilylated by the blending of 6'-N-methyl-3 ', 4'-dideoxycanamycin B. 54. A polysilylated aminoglycoside according to claim 53, characterized in that it contains an average number of silyl groups per cell. molecule of from 3 to 5. The polysilylated aminoglycoside of claim 53 or 54, KENOETEG 56. A polysilylated aminoglycoside according to any one of claims 53-55, characterized in that the amino blocking groups are carbo-benzyloxy or trifluoroacetyl groups. 35 'T.SDOCID <DK 17? S43B>