WO2002070017A1

WO2002070017A1 - Siderophore antibiotic conjugates with tetra- or hexadentate iron chelators on the basis of amino acids or peptides, method for producing them and the use thereof

Info

Publication number: WO2002070017A1
Application number: PCT/EP2002/002074
Authority: WO
Inventors: Steffen Wittmann; Lothar Heinisch; Ute Möllmann
Original assignee: Grünenthal GmbH
Priority date: 2001-03-01
Filing date: 2002-02-27
Publication date: 2002-09-12
Also published as: DE10111163A1

Abstract

The invention relates to novel conjugates of beta-lactam antibiotics of the general formula (I) with tetra- or hexadentate iron chelators or the acylated derivatives thereof as siderophore components, derived from amino acids or peptides, comprising at least 2 catechol substituents. In formula (I), R1 represents a beta-lactam antibiotic, optionally with an intercalated further amino acid moiety, or R1 represents various groups that contain a catechol or hydroxamate group as a complement to a hexadentate chelator structure, optionally in the acylated form or together with spacer groups, and that further contain the group of a beta-lactam antibiotic, especially of a penicillin or cephalosporin derivative, especially ampicillin, amoxicillin or cefaclor (as a free acid, in the form of their salts or their easily cleavable esters). The inventive compounds can be introduced into bacterial cells via the iron transport pathways on account of their siderophore components, thereby efficiently improving or enlarging their activity to a greater extent than known compounds of the kind. The inventive compounds are capable of overcoming resistances of problematic germs that cannot be treated.

Description

Siderophore-antibiotic conjugates with 4- or 6-toothed iron chelators based on amino acids or peptides, process for their preparation and their use

The present invention relates to new siderophore-antibiotic conjugates, wherein 6-toothed iron chelators or their derivatives masked with acyl groups, which are derived from amino acids or peptides, function as siderophore components. With the siderophore components (R ¹ or A = OH) at least 4 chelating groups of catechol units are formed. The new Siderophor-antibiotic conjugates are antibacterial, especially against Gram-negative bacteria, whereby the antibiotics are introduced into the bacterial cell via iron transport routes and can therefore improve or expand their effectiveness with reduced side effects much more than previous compounds of this type contribute to combating penetration-related antibiotic resistance, which plays a central role in the treatment of bacterial infections.

Siderophore-antibiotic conjugates of the formula I with the stated substituents have not hitherto been described in the literature.

It is known that certain catechol structures play an important role in natural siderophores as iron-complexing structural elements ("Iron Transport in Microbes, Plants and Animals", ed .: Winkelmann, G., van Helm, D., Neilands, JB, V.Ch . Publishing house Weinheim, 1987), for example, the enterobactin, a siderophore in E. coli and other bacterial strains, is a trimer of N- (2,3-dihydroxybenzoyl) -L-serine. The monomer is also effective as a siderophore (Hantke, K., FEMS Microbiol. Lett. 67 (1990), 5). The N- (2,3-dihydroxybenzoyl) glycine has been found as a siderophore in B.subtilis (lto, T., Neilands, JB, J.Amer.Chem Soc. 80 (1958), 4645). Some catechol-substituted amino acid derivatives have already been prepared synthetically, for example N- (2,3-dihydroxybenzoyl) -L-threonine (Kanai, F .; Kaneko, T., Morishima, H., Isshiki, K., Takita, T., Takeuchi, T., Umezawa. H., J. Antibiot. 38 (1985), 39), the N ² , N ⁶ bis (2,3-dihydroxybenzoyl) -L-lysine (Corbin, JL, Bulen.WA , Biochemistry 8 (1969), 757; McKee, JA, .Sharma, SK, Miller, MJ; Bioconjugate Chem., 2 (1991) 281) and N ² , N ^6- bis (2,3-dihydroxybenzoyl) -ysyl -N ⁶ - (2,3-dihydroxybenzoy!) Lysine (Chimiak, A., Neilands, JB, Structure and Bonding, 58 (1984), 89). Various O-acylated catechol compounds derived from mono- and diamino acids (L. Heinisch, M. Schnabelrauch, U. Möllmann, R. Reissbrodt, DE 19654920 A1) as well as benzoxazine-2,4-dione derivatives derived from these catechol compounds (L Heinisch, S. Wittmann, U. Möllmann, R. Reissbrodt, EP 0 863 139 A1) have become known. Some derivatives of polybasic secondary amino acids have already been described for the latter compounds. The catechol derivatives mentioned have been converted with antibiotics to conjugates which are highly antibacterial in vitro. Various other catechol compounds have been linked to β-lactams, which has led to a considerable increase in the antibacterial activity of these antibiotics, as a result of the introduction into the bacterial cell via bacterial iron transport routes (for example Arisawa, M., Sekine, Y., Shimizu, S., Takano , H., Angehrn, P., Then, RL, Antimicrob. Agents Chemother. 35 (1991), 653). So far, however, no such compounds have reached clinical maturity. To achieve this goal, further siderophore-antibiotic conjugates with new synthetic siderophores must be sought, which have even better antibacterial activity, e.g. against resistant pathogenic problem germs, such as Stenotrophomonas maltophilia, and fewer side effects than the previously known compounds of this type.

The invention serves to obtain new siderophore-antibiotic conjugates with 4- or 6-toothed iron chelators which contain at least 2 catechol units or their acylated derivatives, derived from amino acids or peptides, and for their use. The aim of the invention is to find suitable compounds for introducing active substances, e.g. of antibiotics in the bacterial cell, which exceed the previously described compounds of this type. The use of acylated catechol derivatives is intended to ensure that the compounds have improved pharmacological properties or can serve as pharmacological transport forms for the catechol compounds which actually promote penetration and are less toxic.

The invention has for its object to find new siderophore-antibiotic conjugates of the general formula I, which have stronger antibacterial activity as comparable known compounds of this type. 4- or 6-toothed iron chelators or their acylated derivatives are said to function as siderophore components, at least 2 catechol substituents or their acylated derivatives, derived from amino acids or peptides, being involved.

The object is achieved according to the invention by providing new siderophore-antibiotic conjugates with 4- or 6-toothed iron chelators or their acylated derivatives based on amino acids or peptides of the general formula I,

Wherein

R ¹ = A,

with R> 6 _ = H, alkyl, hydroxyalkyl, acyloxyalkyl, alkyloxyalkyl, carboxy,

Alkyloxycarbonyl, aryl, arylalkyl, arylalkyloxyalkyl, hetaryl, hetarylalkyl, m = 0-5 = R ¹¹

with o = 1 - 5

with R> 7 _ = H, alkyl, alkyl, alkyloxycarbonylalkyl, R 8 ^ö _ = H, aroyl, o = 1-5

with R ⁸ = H, aroyl, R ⁹ = H, alkyl, cycloalkyl, alkyloxycarbonylalkyl, o = 1-5

= R 15

with o = 1 -5, p = 1 - 5, R ¹⁰ = H, COalkyl, COOalkyl = R ¹⁶

R ² = H, CO-alkyl, COO-alkyl, R ⁵ = H or R ⁵ together with one of the radicals R ² or R ^{10 represents} a group -CO-, R ³ = H, CO-alkyl, COO-alkyl,

R ⁴ = H, alkyl, substituted alkyl, aryl, substituted aryl, halogen, alkoxy, substituted alkoxy, in all possible positions, also occurring several times, A = the residue of a ß-lactam antibiotic, preferably the residue of a penicillin derivative, in particular an ampicillin or amoxicillin residue (formula B) or a bacampicillin residue (α-ethoxycarbonyloxyethyl ester of ampicillin), or the residue of a cephalosporin, in particular a cefachlor residue (formula C), or a quinolone residue

B: R = H: ampicillin residue, R = OH: amoxicillin residue C: cefaclor residue

mean, in the above formulas acyl in particular CrC -alkanoyl or -CC ₄ alkoxycarbonyl, alkyl and alkoxy, also in word combinations such as alkoxycarbonyl, are in particular -C ₈ -alkyl or alkoxy, substituted alkyl for halogen , Alkoxy, hydroxy, carboxy and alkoxycarbonyl substituted alkyl, substituted alkoxy for alkoxy substituted by halogen, alkoxy, carboxy and alkoxycarbonyl, a substituted phenyl is a phenyl substituted by alkyl, halogen, alkoxy, hydroxy, carboxy and alkoxycarbonyl. If asymmetric carbon atoms are present, the corresponding D and L forms, enantiomers and diastereomers and the racemates or mixtures of enantiomers and diastereomers are likewise the subject of the invention. The compounds mentioned can be present as free acids, in the form of their salts or as readily cleavable esters which can be cleaved under physiological conditions, with the proviso that R ⁴ and / or R ^{5 is} not H when R ¹ = A.

The compounds of the formula I provided according to the invention are prepared as follows:

First of all, bis-catechol derivatives of the formula I with R ¹ or A = OH are prepared from corresponding diamino acids and derivatives of 2,3-dihydroxybenzoic acid (for example 2,3-diacyloxybenzoyl chloride) and then with the corresponding amino acid derivatives R ¹ using processes known per se H, where R ¹ = R ¹¹ - R ¹⁶ , implemented by customary methods, for example by the mixed anhydride method or by the active ester method, so that the siderophore components (formula I with R ¹ = R ¹¹ - R ¹⁶ and A = OH) can be obtained. These compounds are then treated with an appropriate antibiotic, in particular with a ß-lactam antibiotic, for example ampicillin or amoxicillin, or a suitable cephalosporin derivative by customary processes, preferably by the anhydride process (for example using isobutyl chloroformate), by the active ester process (for example using N-hydroxysuccinimide and dicyclimidylcarb ) or implemented according to the chloride method to the Siderophor antibiotic conjugates of formula I with A = antibiotic.

The starting compounds of the formula I with R ¹ or A = OH are prepared as follows: First, biscatechol derivatives of the formula I with R ¹ = OH are obtained from corresponding diamino acids and derivatives of 2,3-dihydroxybenzoic acid (for example 2nd , 3-diacyloxybenzoyl chloride) and these are then reacted with the corresponding amino acid derivatives R ¹ H, where R ¹ = R ¹¹ - R ¹⁶ , by customary methods, for example by the mixed anhydride method or by the active ester method, so that the siderophore components of the formula I with R ¹ = R ¹¹ - R ¹⁶ can be obtained.

The compounds of the formula I with a carboxyl group can be present as free acids, in the form of their salts or as easily cleavable esters, in particular cleavable under physiological conditions. The compounds can be further purified by customary methods known from the prior art, for example by means of chromatographic methods.

The compounds of formula I according to the invention show antibacterial activity, some of which the effectiveness of previously known comparable compounds far exceeds.

The antibacterial activity was tested in a microdilution test according to the National Committee for Clinical Laboratory Standards, 1998, Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically, Approved Standard M7-A4, NCCLS, Villanova, Pa minimal inhibitory concentrations (MIC values) tested against the following bacterial strains; Against the Gram-negative strains Pseudomonas aeruginosa SG 137, ATCC 27853, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC 10031, Stenotrophomonas maltophilia GN 12873, Serratia marcescens SG 621 and against the Gram-positive strain Staphylococcus aureus SG 511.

The results of the antibacterial testing are summarized in the table. The corresponding values of azlocillin, ampicillin and meropenem are given for comparison. The results show that the substances shown according to the invention partly. against some bacterial strains far exceed the inhibitory values of the comparison substances and can successfully overcome bacterial resistance. The compounds also show a greater activity against Gram-negative bacteria than hitherto, known corresponding catecholate-β-lactam conjugates according to the literature mentioned above, including against the problem germ Stenotrophomonas maltophilia.

Because of their antibacterial properties, the compounds of the general formula I are suitable for use as medicaments for bacterial infections. In the case of such diseases, the compounds of the formula I can be used either alone or with physiologically tolerable auxiliaries or excipients, it being possible in principle for all customary pharmacological uses and physiologically tolerable dosages.

Examples

Example 1 N-rN ² .N ⁵ -Bis- ( ^, 5-bromo-2,3-diacetoxybenzovπ-L-omithvn-ampicillin

Formula I with R ¹ = Ampicillino, R ² , R ³ = COCH ₃ , R ⁴ = 5-Br, R ⁵ = H, n = 3. To a solution of 0.730 g (1 mmol) N ² , N ⁵ -Bis - (5-bromo-2,3-diacetoxy-benzoyl) -L-omithin and 0.112 ml of N-methylmorpholine in 10 ml of anhydrous tetrahydrofuran were added at -20 ° C with stirring 0.131 ml (1 mmol) of isobutyl chloroformate. The mixture was stirred at about -10 ° C. for one hour and then a solution of 0.412 g (1 mmol) of ampicillin trihydrate and 0.140 ml (1 mmol) of triethylamine in 5 ml of 80% tetrahydrofuran was added. The mixture was stirred for one hour at about -10 ° C. and for one hour at 20 ° C., then evaporated in vacuo, the residue was mixed with ethyl acetate and water and carefully acidified with 1 M hydrochloric acid. After shaking, the organic phase was separated, washed with aqueous sodium chloride solution, dried over sodium sulfate and evaporated. The residue was purified by preparative HPLC on silica gel (Eurospher 100 C18, 7 μm, Knauer, Berlin) with a mixture of acetonitrile / water (37.5 / 62.5) as the eluent. The acetonitrile was distilled off from the corresponding fraction in vacuo and the residue was lyophilized. 0.40 g (40% of theory) of the title compound were obtained in the form of a colorless solid.

¹ H NMR (DMSO-d6): 1, 39 (s, 3H, CH _3); 1, 53 (s, 3H, CH _3); 1.45 - 1.90 (m, 4H, 2 x CH ₂ ); 2.16 (s, 3H, COCH ₃ ); 2.21 (s, 3H, COCH ₃ ); 2.27 (s, 6H, COCH ₃ ); 3.17 (m, 2H, CH _2); 4.19 (s, 1H, 3-CH); 4.62 (1H, m, CH); 5.38 (d, 1H, 7-CH); 5.52 (q, 1H, 6-CH); 5.75 (d, 1H, a-CH); 7.20 - 7.75 (m, 9H, aromat); 8.45 (t, 1H, NHCO); 8.52 (d, 1H, NHCO); 8.67 (d, 1H, NHCO); 9.16 (d, 1H, NHCO);

The sodium salt of the title compound was prepared by adding a solution of 0.02 g of sodium ethyl hexanoate in 3 ml of ethyl acetate to a solution of 0.10 g of the title compound in 12 ml of ethyl acetate. The precipitate which had separated out was filtered off after standing for 10 minutes and washed with petroleum ether. The sodium salt of the title compound was obtained in the form of a colorless amorphous solid in 90% yield. Example 2

N- (N'-rN ² .N ⁵ -Bis- (2,3-diacetoxybenzovπ-L-ornithyll-L-0-benzyl-seryl) -ampicillin

Formula I with R ¹ = R ¹¹ , R ⁴ , R ⁵ = H; R ² , R ³ = COCH ₃ , R ⁶ = CH ₂ 0 benzyl, n = 3, m = 0,

A = Ampicillino. The title compound or its sodium salt was prepared analogously to

Example 1 from N- [N ² , N ⁵ -Bis- (2,3-diacetoxybenzoyl) -L-ornithyl] -LO-benzyl-serine and

Ampicillin trihydrate in 50% yield in the form of a colorless amorphous

Solid.

¹ H NMR (DMSO-d6): 1.40 (s, 3H, CH ₃ ); 1, 55 (s, 3H, CH _3); 1 45 -1 90 (m, 4H, .2 x CH _2); 2.19 (s, 3H, COCH ₃ ); 2.20 (s, 3H, COCH3); 2.27 (s, 6H, COCH ₃ ); 3.18 (m, 2H,

CH ₂ ); 3.63 (m, 2H, CH _2); 4.19 (s, 1H, 3-CH); 4.47 (2H, s, CH ₂ Ph); 4.55 (m, 1H, CH);

); 4.78 (1H, m, CH); 5.38 (d, 1H, 7-CH); 5.51 (q, 1H, 6-CH); 5.78 (d, 1H, a-CH); 7.20

- 7.60- (m, 16H, aromat); 8.18 (d, 1H, NHCO); 8.33 (m, 2H, 2xNHCO); 8.68 (d, 1H,

NHCO); 9.20 (d, 1H, NH).

Example 3 Ni ^" N-rN ² , N ^5- bis (2,3-diacetoxybenzovπ-L-ornithvn-L-tryptophanyll-ampicillin Formula I with R ¹ = R ¹¹ , R ² , R ³ = COCH ₃ , R ⁴ , R ⁵ = H., R ⁶ = L-indolyl-2-methyl, n = 3, m = 0, L, L-form A = ampicillino The title compound or its sodium salt was prepared analogously to

Example 1 from N- [N ² , N ^5- bis (2,3-diacetoxybenzoyl) - -ornithyl] -L-tryptophan and

Ampicillin trihydrate in 30% yield in the form of a colorless amorphous

Solid.

¹ H NMR (DMSO-d6): 1, 39 (s, 3H, CH _3); 1, 54 (s, 3H, CH _3); 1.50-1.90 (m, 4H, 2x CH ₂ ); 2.16 (s, 3H, COCH3); 2.17 (s, 3H, COCH3); 2.27 (s, 6H, COCH ₃ ); 3.18 (m, 2H, NCH _2); 3.35 (m, 2H, _Tr-CH2); 4.18 (s, 1H, 3-CH); 4.44 (m, 1H, CH); 4.81 (, 1H, CH); 5.36 (d, 1H, 7-CH); 5.49 (q, 1H, 6-CH); 5.72 (d, 1H, a-CH); 6.85 - 7.60 (m, 16H, aromat.); 8.07 (d, 1H, NHCO); 8.32 (m, 2H, 2xNHCO); 8.71 (d, 1H, NHCO); 9.20 (d, 1H, NHCO); 10.78 (s, 1H, NHCO).

Example 4 N- (N'-rBis-N, N ⁵ - (2,3-diacetoxybenzoyl) -L-ornithyll-L-phenylalanyl) -ampicillin Formula I with R ¹ = R ¹¹ , with R ⁶ = benzyl, R ² , R ³ = COCH ₃ , R ⁴ , R ⁵ = H, n = 3, m = 0, A = Ampicillino. The title compound or its sodium salt was prepared analogously to Example 1 from N- [bis-N ² , N ⁵ - (2,3-diacetoxybenzoyl) -ornithyl] -L-phenylalanine and ampicillin trihydrate in the form of a 30% yield colorless amorphous solid. ¹ H NMR (DMSO-d6): 1.40 (s, 3H, CH ₃ ); 1, 55 (s, 3H, CH _3); 1.50 -1.90 (m, 4H, 2x CH ₂ ); 2.18 (s, 3H, COCH ₃ ); 2.19 (s, 3H, COCH ₃ ); 2.27 (s, 6H, COCH ₃ ); 2.99 (m, 2H, CH ₂ -Ph); 3.17 (m, 2H, CH _2); 4.18 (s, 1H, 3-CH); 4.45 (m, 1H, CH); 4.82 (m, 1H, CH); 5.37 (d, 1H, 7-CH); 5.52 (q, 1H, 6-CH); 5.78 (d, 1H, a-CH); 7.10 - 7.50 (m, 16H, aromatic H); 8.10 (d, 1H, NHCO); 8.21 (t, 1H, NHCO); 8.30 (d, 1H, NHCO); 8.85 (d, 1H, NHCO); 9.22 (d, 1H, NHCO).

Example 5 N- (2-rN ² , N ⁵ -Bis- (2,3-diacetoxybenzoyl) -L-ornithyl-aminol-4- (benzyloxycarbonvπ-n-butanovD-ampicillin Formula I with R = R ¹¹ , R ² , R ³ = COCH ₃ , R ⁴ , R ⁵ = H., R ⁶ = 2-benzyloxycarbonylethyl, n = 3, m = 0, L, L-form, A = Ampicillino.

The title compound or its sodium salt was prepared analogously to Example 1 from N- [N ² , N ^5- bis (2,3-diacetoxybenzoyl) -L-ornithyl] -L-glutamic acid 5-benzyl ester and ampicillin trihydrate in 45 % yield in the form of a colorless amorphous solid.

¹ H NMR (DMSO-d6): 1, 39 (s, 3H, CH _3); 1, 54 (s, 3H, CH _3); 1.50 -1.90 (m, 6H, 3 x CH ₂ ); 2.18 (s, 3H, COCH3); 2.19 (s, 3H, COCH3); 2.26 (s, 6H, COCH ₃ ); 2.32 (m, 2H, CH _2); 3.17 (m, 2H, CH _2); 4.19 (s, 1H, 3-CH); 4.48 (m, 2H, 2xCH); 5.04 (s, 2H, benzyl. CH ₂ ); 5.37 (d, 1H, 7-CH); 5.51 (q, 1H, 6-CH); 5.73 (d, 1H, a-CH); 7.25 - 7.50 (m, 16H, aromat.); 8.10 (d, 1H, NHCO); 8.29 (m, 1H, NHCO); 8.35 (d, 1H, NHCO); 8.82 (d, 1H, NHCO); 9.17 (d, 1H, NHCO).

Example 6 Nf N'.N "N '" - tris (2,3-diacetoxybenzoyl) -N ² -L-lvsyllvsyl} -ampicillin sodium salt formula I where R ¹ = R ^12, R ^2, R ³ = COCH _3, R ⁴ , R ⁵ = H., n = 4, L-shape, o = 4, L, L-shape, A = Ampicillino.

The title compound was prepared analogously to Example 1 from N ', N "N"' - tris (2,3-diacetoxybenzoyl) -N ² -L-lysyl-L-lysine and ampicillin trihydrate in 70% yield in the form of a colorless amorphous solid. ¹ H NMR (DMSO-d6): 1, 38 (s, 3H, CH _3); 1, 51 (s, 3H, CH _3); 1, 20 -1, 80 (m, 12H, 6 x CH ₂ ); 2.21 (s, 9H, COCH ₃ ); 2.29 (s, 9H, COCH3); 3.13 (m, 4H, 2xNCH _2); 3.80 (s, 1H, 3-CH); 4.43 (m, 2H, 2xCH); 5.23 (m, 1H, 7-CH); 5.34 (m, 1H, 6-CH); 5.75 (m, 1H, a-CH); 7.20 - 7.55 (m, 14H, aromat); 8.00 - 9.00 (m, 6H, δxNHCO).

Example 7 Nf N'.N "N '" - tris (2,3-diacetoxybenzovn-N ² -L-lvsyl-L-lvsyl) amoxicillin Formula I with R ¹ = R ¹² , R ² , R ³ = COCH3 , R ⁴ , R ⁵ = H., n = 4, L-form, o = 4, L, L-form, A = Amoxicillino. The title compound or its sodium salt was prepared analogously to Example 1 from tris-N ', N "N"' - (2,3-diacetoxybenzoyl) -N ² -L-lysyl-L-lysine and amoxicillin trihydrate in 50% strength Yield in the form of a colorless amorphous solid. ¹ H NMR (DMSO-d6): 1, 38 (s, 3H, CH _3); 1, 50 (s, 3H, CH _3); 1, 20 -1, 80 (m, 12H, 6 x CH ₂ ); 2.21 (s, 9H, COCH3); 2.29 (s, 9H, COCH ₃ ); 3.13 (m, 4H, 2xNCH _2); 3.83 (s, 1H, 3-CH); 4.44 (m, 2H, 2xCH); 5.25 (m, 1H, 7-CH); 5.34 (m, 1H, 6-CH); 5.75 (m, 1H, a-CH); 6.65 - 7.55- (m, 13H, aromat); 8.00 - 9.00 (, 7H, 6xNHCO, OH).

Example 8 N- { ^" 2- {rN ', N" -Bis- (2,3-diacetoxybenzovπ-D-ornithyll-amino-4-r (N-benzoyloxy-N-methvO-aminoyπ-L-butanoyl) -ampicillin

Formula I with R ¹ = R ¹⁴ ; R ² , R ³ = COCH3, R ⁴ , R ⁵ = H., n = 3, D-form, R ⁸ = benzoyl,

R ⁹ = CH _3, o = 2, L-form, A = Ampicillino.

The title compound or its sodium salt was prepared analogously to Example 1 from N- [N ', N "-bis (2,3-diacetoxybenzoyl) -D-omithyl] -L-glutamic acid-5- (N'-benzoyloxy-N '-methyl) -amide and ampicillin trihydrate in 40% yield in the form of a colorless amorphous solid.

¹ H NMR (DMSO-d6) 1, 39 (s, 3H, CH _3); 1, 53 (s, 3H, CH _3); 1 45 -2.06 (m, 6H, CH _2);

2.18 (s, 6H, COCH ₃ ); 2.27 (s, 6H, COCH3); 2.39 (m, 2H, CH _2); 3.15 (m, 2H, CH _2); 3.26 (m, 3H, NCH _3); 4.18 (s, 1H, 3-CH); 4.49 (m, 2H, 2xCH); 5.35 (d, 1H, 7-CH);

5.49 (q, 1H, 6-CH); 5.73 (d, 1H, a-CH); 7.20 - 7.45- (m, 11H, aromat); 7.54 (t, 2H, aromat); 7.75 (t, 1H, aromat); 8.00 (d, 2H, aromat); 8.21 (d, 2H, 2xNHCO); 8.32

(m, 1H, NHCO); 8.65 (d, 1H, NHCO); 9.18 (d, 1H, NHCO). Example 9 Ni ^r 2-iTN ² , N ^6- bis (2,3-diacetoxybenzoyl) -L-lvsvn-amino -4-r (N-benzoyloxy-N-methvπ-aminovπ-L-butanoyl) -ampicillin

Formula I with R ¹ = R ¹⁴ , R ² , R ³ = COCH ₃ , R ⁴ , R ⁵ = H., n = 4, L-form, R ⁸ = benzoyl, R ⁹ = CH _3, o = 2, L-shape, A = ampicillino.

The title compound or its sodium salt was prepared analogously to Example 1 from N- [N ² , N ⁶ -bis (2,3-diacetoxybenzoyl) -L-lysyl] -L-glutamic acid-5- (N'-benzoyloxy-N '-methyl) -amide and ampicillin trihydrate in 20% yield in the form of a colorless amorphous solid. ¹ H NMR (DMSO-d6): 1, 39 (s, 3H, CH _3); 1, 53 (s, 3H, CH _3); 1, 45 to 2.06 - (m, 8H, CH _2); 2.19 (s, 3H, COCH ₃ ); 2.21 (s, 3H, COCH ₃ ); 2.27 (s, 6H, COCH ₃ ); 2.39 (m, 2H, CH _2); 3.15 (m, 2H, CH _2); 3.30 (m, 3H, NCH _3); 4.19 (s, 1H, 3-CH); 4.38 (m, 1H, CH); 4.46 (m, 1H, CH); 5.35 (d, 1H, 7-CH); 5.50 (q, 1H, 6-CH); 5.73 (d, 1H, a-CH); 7.25 - 7.60 (m, 13H, aromat); 7.74 (t, 1H, aromatic H); 7.97 (d, 2H, aromat); 8.10 (m, H, NHCO); 8.30 (m, 2H, 2xNHCO); 8.55 (d, 1H, NHCO; 9.17 (d, 1H, NHCO).

Example 10 Ni ^r 2-rN ² , N ⁵ 'bis (2,3-diacetoxybenzoyl) -D-omithvπ-amino1-4-r (N-benzoyloxy-N-cvclohexyl) amidovn-L-butanoyl) -ampicillin formula I with R ¹ = R ¹⁴ , R ² , R ³ = COCH ₃ , R ⁴ , R ⁵ = H, n = 3, D-form, R ⁸ = benzoyl, R ⁹ = cyclohexyl, o = 2, L-Fόrm , A = Ampicillino.

The title compound or its sodium salt was prepared analogously to Example 1 from N- (N ² , N ⁵ -bis- (2,3-diacetoxybenzoyl) -D-omithyl) -L-glutamic acid-5- (N'-benzoyloxy- N'-cyclohexyl) -amide and ampicillin trihydrate in 50% yield in the form of a colorless amorphous solid.

¹ H NMR (DMSO-d6): 1, 38 (s, 3H, CH _3); 1, 53 (s, 3H, CH _3); 1.00 - 2.30 (m, 18H, 10 x CH ₂ ); 2.18 (s, 6H, COCH ₃ ); 2.27 (s, 6H, COCH ₃ ); 3.15 (m, 2H, NCH _2); 4.18 (s, 1H, 3-CH); 4.20 (m, 1H, CH); 4.48 (m, 2H, 2xCH); 5.35 (d, 1H, 7-CH); 5.48 (q, 1H, 6-CH); 5.74 (d, 1H, a-CH); 7.22 - 8.05- (m, 16H, aromat); 8.22 (d, 2H, 2xNHCO); 8.32 (m, 1H, NHCO); 8.65 (d, 1H, NHCO; 9.19 (d, 1H, NHCO).

Example 11 N-fN ² -r4- (N'-benzoyl-N'-methvn-amido-qlutarovn-N ⁶ - (N ² .N ⁶ -bis-2.3- diacetoxybenzoyl) -L-lvsvn-L-lvsyl) - ampicillin Formula I with R ¹ = R ¹⁵ , R ² , R ³ = COCH ₃ , R ⁴ , R ⁵ = H., n = 4, L-form, R ⁸ = benzoyl,, R ⁹ = CH _3, p = 4 , q = 3, A = Ampicillino.

The title compound or its sodium salt was prepared analogously to Example 1 from N ² - [(N'-benzoyloxy-N'-methyl-amido-glutaroyl) -N ⁶ - (N ² , N ⁵ -bis-2,3- diacetoxybenzoyl) -L-lysyl] -L-lysine and ampicillin trihydrate in 30% yield in the form of a colorless amorphous solid.

¹ H NMR (DMSO-d6): 1, 39 (s, 3H, CH _3); 1, 53 (s, 3H, CH _3); 1, 20-2.40 (m, 18H, 4 x CH ₂ ); 2.19 (s, 3H, COCH3); 2.22 (s, 3H, COCH ₃ ); 2.27 (s, 6H, COCH3); 3.10 (m, 4H, 2xNCH ₂ ); 4.19 (s, 1H, 3-CH); 4.33 (m, 2H, 2xCH); 5.37 (d, 1H, 7-CH); 5.50 (q, 1H, 6-CH); 5.68 (d, 1H, a-CH); 7.30 -8.05 (m, 18H, aromat. + 2x NHCO); 8.20 (d, 1H, NHCO); 8.29 (m, 1H, NHCO); 8.40 (m, 1H, NHCO); 9.10 (m, 1H, NHCO).

Example 12 NN ² -r (N'-benzoyloxy-N'-cvclohexyl-amido-qlutarovπ-N ⁶ -rN ^{2 '} .N ^6' -bis- (2.3-diacetoxybenzoyl) -L-lvsvπ-L-Ivsyl) -ampicillin

Formula I with R ¹ = R ¹⁵ , R ² , R ³ = COCH3, R ⁴ , R ⁵ = H., n = 4, L-form, R ⁸ = benzoyl, R ⁹ = cyclohexyl _, p = 4, q = 3, A = Ampicillino.

The title compound or its sodium salt was prepared analogously to Example 1 from N ² - [(N ^, -benzoyloxy-N'-cyclohexyl-amido-glutaroyl] -N ⁶ - [N ² , N ^{6 '} -bis- (2, 3-diacetoxybenzoyl) -L-lysyl] - L-lysine and ampicillin trihydrate in 50% yield in the form of a colorless amorphous solid

¹ H NMR (DMSO-d6): 1, 32 (s, 3H, CH _3); 1, 54 (s, 3H, CH _3); 1.00 - 2.30 (m, 28H, 8 x CH ₂ ); 2.19 (s, 3H, COCH3); 2.22 (s, 3H, COCH ₃ ); 2.27 (s, 6H, COCH ₃ ); 3.03 (m, 2H, NCH _2); 3.15 (m, 2H, NCH _2); 4.33 (m, 3H, 3xCH); 5.37 (d, 1H, 7-CH); 5.51 (q, 1H, 6-CH); 5.68 (m, 1H, a-CH); 7.20 - 7.95 (m, 16H, aromat.); 8.04 (d, 2H, 2xNHCO); 8.29 (m, 1H, NHCO); 8.45 (m, 1H, NHCO); 9.10 (m, 1H, NHCO).

Example 13 N- {N ² -rε- (8-methoxycarbonyloxy-3,4-dihydro-2,4-dioxo-2H-1,3-benzoxazin-3-yl) -n-hexanovn-N ⁶ - [2,6- bis- (8-methoxycarbonyloxy-3.4-dihvdro-2,4-dioxo-2 - / - 1.3-benzoxazin-3-yl) -n-hexanovπ-L-lvsvP-am ^p icillin Formula I with R ¹ = R ¹⁶ , R ² and R ¹⁰ = -CO- in combination with R ⁵ , R ³ = COOCH ₃ , R ⁴ = H, R ⁵ = -CO- in combination with R ² and R ¹⁰ , n = 4, L-shape, o = 4, p = 5, A = Ampicillino.

The title compound or its sodium salt was prepared analogously to Example 1 from N ² - [ε- (8-methoxycarbonyloxy-3,4-dihydro-2,4-dioxo-2H-1,3-benzoxoxazin-3-yl ) -n-hexanoyl] -N ⁶ - [2,6-bis- (8-methoxycarbonyloxy-3,4-dihydro-2,4-dioxo-2H-1,3-benzoxazin-3-yl) -n-hexanoyl ] -L-lysine and ampicillin trihydrate in 30% yield in the form of a colorless amorphous solid.

¹ H NMR (DMSO-d6): 1, 39 (s, 3H, CH _3); 1, 53 (s, 3H, CH _3); 1, 00 to 2.30 (m, 20H, CH _2); 2.97 (m, 2H, NCH _2); 3.79 (m, 4H, NCH _2); 3.90 (s, 9H, COCH ₃ ); 4.19 (s, 1H, 3-CH); 4.25 (m, 1H, CH); 5.07 (m, 1H, CH); 5.37 (d, 1H, 7-CH); 5.50 (q, 1H, 6-CH); 5.65 (m, 1H, a-CH); 7.20 - 8.02- (m, 16H, aromat + NHCO); 8.45 (m, 1H, NHCO); 9.10 (m, 1H, NHCO).

Example 14 ^[ N- (2-rN ² , N ⁶ -Bis- (2,3-diacetoxybenzovπ-L-lvsvπ-aminol-4-r (N-benzoyloxy-N-cyclohexyO-amidovπ-L-butanovD-ampicillin

Formula I with R ¹ = R ¹⁴ , R ² , R ³ = COCH3, R ⁴ , R ⁵ = H., n = 4, L-form, R ⁸ = benzoyl,

R ⁹ = cyclohexyl, o = 2, L-form, A = ampicillino.

The title compound or its sodium salt was prepared analogously to Example 1 from N- (N ² , N ⁶ -bis- (2,3-diacetoxybenzoyl) -L-lysyl) -L-glutamic acid-5- (N'-benzoyloxy- N'-cyclohexyl) -amide and ampicillin trihydrate in 50% yield in

Form of a colorless amorphous solid.

¹ H NMR (DMSO-d6): 1, 39 (s, 3H, CH _3); 1, 53 (s, 3H, CH _3); 1, 00 - 2.30 - (m, 16H, 8 x

CH ₂ ); 2.19 (s, 3H, COCH ₃ ); 2.22 (s, 3H, COCH ₃ ); 2.27 (s, 6H, COCH ₃ ); 3.14 (m, 2H, NCH _2); 4.18 (s, 1H, 3-CH); 4.20 (m, 1H, CH); 4.26 (m, 1H, CH); 4.40 (m, 1H, CH);

5.36 (d, 1H, 7-CH); 5.48 (q, 1H, 6-CH); 5.72 (d, 1H, α-CH); 7.20 - 8.03- (m, 16H, aromat); 8.08 (d, 1H, NHCO); 8.30 (m, 2H, 2x NHCO); 8.53 (d, 1H, NHCO);

9.18 (d, 1H, NHCO). Table. Antibacterial activity of the siderophore-antibiotic conjugates MIC values [in μg / ml]

Claims

claims

1. Siderophore-antibiotic conjugates of the general formula

wherein

R ¹ = A,

or R =

with R ⁶ = H, alkyl, hydroxyalkyl, acyloxyalkyl, alkyloxyalkyl, carboxy, alkyloxycarbonyl, aryl, arylalkyl, arylalkyloxyalkyl, hetaryl, hetarylalkyl m = 0-5 = R ¹¹

with o = 1 - 5

with R ⁷ = H, alkyl, alkyl, alkyloxycarbonylalkyl, R ⁸ = H, aroyl, o = 1-5

with p = 1 -5, q = 1 -5 _ _D 15

R ² = H, CO alkyl, COO alkyl,

R ⁵ = H or R ⁵ together with one of the radicals R ² or R ^{10 represents} a group -CO-,

R ³ = H, CO alkyl, COO alkyl,

R ⁴ = H, alkyl, substituted alkyl, aryl, substituted aryl, halogen, alkoxy, substituted alkoxy, in all possible positions, also occurring several times,

A = the residue of a ß-lactam antibiotic, preferably the residue of a penicillin derivative, in particular an ampicillin or amoxicillin residue (formula B) or a bacampicillin residue (α-ethoxycarbonyloxyethyl ester of ampicillin), or the residue of a cephalosporin, in particular a cefachlor residue (formula C), or a quinolone residue ,.

B: R = H: ampicillin residue, R = OH: amoxicillin residue C: cefaclor residue mean, if asymmetric C atoms are present, the corresponding D and L forms, enantiomers and diastereomers as well as the racemates or enantiomer and diastereomer mixtures of compounds of Formula I, wherein the compounds of formula I can be present as free acids, in the form of their salts or as easily cleavable esters which can be cleaved under physiological conditions, with the proviso that R ⁴ and / or R ^{5 is} not H when R ¹ = A.

2. Compounds of formula I according to claim 1, wherein R ¹ = A, R ¹¹ , R ¹³ or R ¹⁴ and R ² and / or R ³ = H, COalkyl or COOalkyl, R ⁴ = H or halogen, also occurring repeatedly , R ⁵ = H.

3. Compounds of formula I according to claim 1, wherein R ¹ = A and R ² and / or R ³ = H, COalkyl or COOalkyl, R ⁴ = halogen, also occurring several times; R ⁵ = H and A = an ampicillin or amoxicillin residue.

4. Compounds of formula I according to claim 1, wherein R ¹ = R ¹¹ , R ² and / or R ³ = H, COalkyl or COOalkyl, R ⁴ = H or halogen, also occurring repeatedly, R ⁵ = H and R ⁶ = H, alkyl, hydroxyalkyl, alkyloxyalkyl, acyloxyalkyl, carboxy, alkyloxycarbonyl, aryl, arylalkyl, arylalkyloxyalkyl, hetarylalkyl, m = 0-5 and A = an ampicillin or amoxicillin radical.

5. Compounds of formula I according to claim 1, wherein R ¹ = R ¹⁴ , R ² and / or R ³ = H, COalkyl or COOalkyl, R ⁴ = H or halogen, R ⁵ = H, R ⁸ = H or aroyl. and / or R ⁹ = H, alkyl, cycloalkyl, alkylcarbonyloxyalkyl and A = an ampicillin or amoxicillin radical.

6. Compounds of formula I according to claim 1, wherein R ¹ = R ¹⁵ , R ² and / or R ³ = H, COalkyl or COOalkyl, R ⁴ = H or halogen, R ⁵ = H, R ⁸ = H or aroyl and / or R ⁹ = H, alkyl, cycloalkyl, alkylcarbonyloxyalkyl.

7. Use of the compounds of formula I according to claims 1-6 as agents against bacterial infections, in particular against infections with Gram-negative bacteria.

8. Medicament containing a compound of formula I according to claims 1-6 together with conventional carrier materials.