MXPA98001743A - New derivatives of benzoxacindiona, procedure for its preparation and its emp - Google Patents

Abstract

The invention relates to novel benzoxazidene derivatives of the general formula I, and to their conjugates with active substances, for example antibiotics. The compounds according to the invention represent protected catechol derivatives in heterocyclic form and are effective as siderophores in the case of strains of gram-negative bacteria, especially pseudomonates, E. coli and salmonella strains. In the form of their conjugates with active substances, for example antibiotics (such as "siderophore-antibiotic conjugates"), they can introduce these into the bacterial cells and improve or extend their antibacterial effectiveness, partly also with respect to other bacterial strains resistant to la-lactam. . In addition, the aforementioned compounds are suitable as potential prodrug factors for chelators deriving from iron metabolism. The application of the invention can be carried out in pharmaceutical research, industry and agronomy. Formula (I): where R 1 = H or carboxyalkyl, R 2 = H, alkyl or phenyl, and R 3 = various acid groups derived from amino acids, dipeptides and hydrozones, or their conjugates with active substances, for example antibiotics. The indicated compounds can be present as free acids, in the form of their salts or asosteres easily unfolded

Description

NEW DERIVATIVES OF BENZOXACINDIONA. PROCEDURE FOR YOUR PREPARATION AND YOUR EMPLOYMENT.

Description of the Invention The present invention relates to new benzoxazidene derivatives, as well as to their conjugates with active substances, for example antibiotics. The compounds are effective as siderophores in the case of gram-negative bacteria, they can specifically supply iron ions to the bacteria and, in the form of their conjugates with active substances, for example antibiotics (such as "siderophore-antibiotic conjugates"), they can introduce these into the bacterial cell through iron transport routes, with which they improve or extend the effectiveness of the active substances. This is intended to provide a contribution to combat resistance to antibiotics referred to penetration. The compounds according to the invention represent preliminary phases for iron chelators or catecholic compounds protected in heterocyclic form, that is to say they can be derived enzymatically in corresponding catechol derivatives. In this way they can influence iron metabolism and related diseases in different ways. The industrial field of application of the invention is pharmaceutical research and industry. The benzoxazidene derivatives of the formula I with the indicated substituents have not been described to date in the literature. The compounds in principle represent acylated catecholic derivatives, whose acyl component is bound in a benzoxacin ring. It is known that certain catechoic structures play an important role in natural siderophores as structural elements in iron complexes ("Iron Transport in Microbes, Plants and Animáis", published by Winkelmann, G., van Helm, G., Neilands, JB, Editorial V.Ch.-Verlagsgesellschaf Weinheim, 1987), for example, enterobactin, a siderophore in E. coli and other bacterial strains, is a trimer of N- (2,3-dihydroxybenzoyl) -L-serine. The monomer is also active as siderophore (Hantke, K., FEMS Microbiol. Lett. £ 7 (1990), 5). The N- (2,3-dihydroxybenzoyl) -L-serine was found as siderophore in B. subtilis (Ito, T., Neilands, J.B., J. Amer. Chem. Soc. SQ. (1958), 4645). A number of substituted amino acid derivatives of catechol 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, 3_a (1985), 39), N2, N6-bis- (dihydroxybenzoyl) -L-lysine (Corbin, JL, Bulen, WA, Biochemistry 8. (1969), 757; McKee, JA, Sharma, SK, Miller, MJ, Bioconjugate Chem., 2 (1991) 281) and N2, N6-bis- (2,3-dihydroxybenzoyl) -lysil -Ns- (2,3-dihydroxybenzoyl) -lysine (Chimiak, A., Neilands, JB, Structure and Bonding, 53. (1984), 89). It was further discovered that in various bacterial strains, certain benzohydrazones of glyoxylic acid and certain oxanilic acid derivatives may be used as siderophores (Reissbrodt, R., Heinisch, L., Mollmann, U., Rabsch, W., Ulbricht, H ., BioMetals, 6. (1993), 155). Various catechol compounds were coupled with β-lactams, whereby a considerable increase in the antibacterial activity of these antibiotics was achieved, achieved due to an introduction in the bacterial cell through iron transport routes (for example Arisawa, M ., Sekine, Y., Shimizu, S., Takano, H., Angehrn, P., Then, RL, Antimicrob, Agents Chemother, 3J (1991), 653). However, to date, none of this type of compounds has matured for clinical application. To achieve this goal it is necessary to look for other new synthetic siderophores that are suitable for the formation of conjugates with antibiotics. On the other hand, siderophores as iron chelators have the potential to influence in various ways the biological metabolism of iron and related diseases. A) Yes, for example, the siderophore Desferrioxamine B (Desferal) is successfully applied in diseases that result from an iron overload (for example thalassemia). Benzoxacidene derivatives without a substituent at position 8 are known, which are not derived from catechol, for example 3-carboxymethyl-2,4-dioxobenzoxacin (for example, Lespagnol, A., Lespagnol, Ch., Bernier, JL, Cazin , JC, Cazin, M., Bull, Soc. Pharm, Lille 4, (1972), 179-185). A preparation of benzoxacin derivatives of acyloxybenzoylchloride and amino components with evolution of HCl and methanol has not been described so far. The invention serves to find new benzoxazidene derivatives, as well as for their application. The invention seeks to develop suitable compounds for introducing active substances, for example antibiotics, into the bacterial cell through bacterial routes of iron transport. On the other hand, new preliminary phases must be found with these compounds or forms of prodrugs for iron chelators that can influence in various ways the biological metabolism of iron and the diseases derived from it. The coupling of the cathcolic structure in the heterocyclic structure of benzoxacin is intended to ensure that the compounds in their acylated form, especially their conjugates with antibiotics, have better pharmacological properties compared to free catechols, or that they can serve as forms Pharmacological transportation for catechol compounds that specifically promote penetration. The object of the invention is to find new benzoxazide derivatives that can function as siderophores or biological chelators of iron. According to the invention, the task is solved by providing new benzoxazidene derivatives of the general formula I, wherein R1 = H, COalkyl or COOalkyl, R2 = H, alkyl, halogen, and RJ represents the following substituents: a) R3 = -Z-CHR4-COR5 with Z = with R = H, alkyl, phenyl or substituted phenyl, especially hydroxyphenyl or acyloxyphenyl, or with R * 1 = (CH-) rC0X with X = OA, wherein A = H, alkyl, an alkali metal ion or an ion of ammonium, either it is a substituted ammonium ion, or with X = a radical of active substance, in epsecial a radical of an antibiotic, which is linked through an OH group or an NH group, and with n = 1 -10, or with R = (CH2) nY, where Y represents a radical of bezoxacosene of the form where R1 and R = are as defined above, and both bezoxacidene radicals may be the same or different, and may be n = 1 -10, or with RJ = OA, where A is defined as above, or with R = a radical of active substance, in particular a radical of an antibiotic, which is linked through an OH group or an NH group, or with R5 = NH-CHR8-COR9, with R8 = H, alkyl, phenyl or substituted phenyl and with R = OA, wherein A is defined as above, or with R = a radical of active substance, especially a radical of an antibiotic, which is linked through a group OH or an NH group, = R17 with X and Y as above, n = 1 - 10 and with R6 = H, alkyl, halogen and with R = H, acyl and n = 1 -10 and m = 1-2, or RJ CHR4-COR5 with R and R as above, or = R 18 with R and / or RH, alkyl, phenyl or substituted phenyl, n = 1-10, and with COR9 and R12 in all possible positions, R defined as above and R12 = H, alkyl, halogen, hydroxy, alkoxy , a radical Y of benzoxacose, or with R, R as above, R14, R15 as R1, R2 and can be n = 1 - 10, or d) R3 = = R with R, 13 and COR in all possible positions, and with R 1 H, alkyl, halogen, hydroxy, alkoxy or a Y radical of benzoxazdione, and R as above and p 0 2, or = R or RJ = R22 with R9 as above, R1_6 = H, alkyl, phenyl or substituted phenyl, of) R = a radical of active substance, especially a radical of an antibiotic, which is linked through an OH group or a group NH In the preceding formulas and in the following, acyl means in particular straight or branched chain C ^^ - Cg alkanoyl or straight or branched chain C- ^ Cg alkoxycarbonyl; straight or branched chain alkyl and straight-chain or branched alkoxy, also in word combinations such as straight-chain or branched alkoxycarbonyl, in particular C 1 -C 8 -alkyl or C 2 -C 6 -alkoxy, - a substituted phenyl means a phenyl more substituted by straight or branched chain alkyl, halogen, especially Cl or F, straight or branched chain alkoxy, hydroxy, carboxy, straight or branched chain alkoxycarbonyl and substituted alkyl of halogen. A substituted ammonium is, for example, an ammonium substituted one or more times of alkyl, as one to four times. An alkali metal ion can be, for example, a sodium or potassium ion. An active substance means, for example, the radical of a suitable antibacterial active substance of any type with free NH or OH group, whereby the active substance is esterified or amidated with the compound of formula 1 through this NH or OH group. The link between the catechol derivative and the antibiotic can be performed both directly and also by customary binding groups, for example aminocarboxylic acids, hydroxycarboxylic acids, diamines or diols. By antibiotic is meant a corresponding β-lactam containing an NH or OH group, such as for example a cephalosporin, for example cephalexin, cefadroxil or clarofan, or a penicillin, for example ampicillin, amoxicillin, or an O-acyl derivative of amoxicillin, or a tetracycline derivative, for example aminodoxycycline, or an antibiotic of the type of aminoglycosides, macrolides, quinolones or carbapenems. In the case of the presence of asymmetric carbon atoms, the object of the invention is also the corresponding Right and Left forms, enantiomers and diastereomers, as well as racemates or mixtures of enantiomers and diastereomers. The aforementioned compounds can be present as free acids, in the form of their salts, or as easily dissociated esters, as required under physiological conditions. The compounds of formula I prepared according to the invention with R 5 or R 9 = OH are prepared by condensation of 2,3-diazyloxybenzoylchloride with the corresponding amino components. For example amino acids, dipeptides, or aminobenzoic acids in sodium carbonate solution, according to formulation scheme 1 indicated: with R 1 = COOalkyl or H Scheme of formulation 1 During the course of the reaction, the COOalkyl group can be split hydrolytically into R, so that compounds can be obtained with R = H. These can be acylated again, for example to compounds with R - Alkyl. The compounds of the formula I with R 5 or R 9 OH can be prepared according to the same principle also according to reaction scheme 2, proceeding in such a way that the corresponding amide derivatives are reacted, for example 2, 3-dihydroxybenzhydrazone , with alkyl esters of chloroformic acid in an alkaline medium.

Formulation scheme 2 The compounds of the formula I according to the invention with R3, R5, R9 or X = a radical of an active substance, for example the radical of an antibiotic, are prepared, for example, by proceeding in such a way that to. The corresponding chloride is first prepared from a compound of the formula I with A = H following conventional procedures, for example by phosphorus pentachloride in carbon tetrachloride, the chloride then being reacted with an active substance or antibiotic, which contains an OH or free NH group as well as optionally a common binding group, such as for example residues of a diaminocarboxylic acid, a hydrocarboxylic acid or a diamine or a diol, in a suitable solvent, for example tetrahydrofuran, or proceeding in a manner what B. a compound of the formula I is reacted with A = H according to the mixing method of mixed anhydrides, first with chloroformic acid ester and a tertiary amine, for example triethylamine, and then together with a suitable tertiary amine, for example triethylamine , with the corresponding active substance containing a free NH or OH group, as well as optionally a common linking group, such as, for example, residues of a diaminocarboxylic acid, of a hydrocarboxylic acid or of a diamine or of a diol, in a suitable solvent , for example tetrahydrofuran. The compounds of formula I with a carboxyl group can be present as free acids, in the form of their salts, or as easily dissociated esters, in particular unfolding under physiological conditions. The purification of the compounds is carried out according to the usual procedures known from the state of the art, for example by recrystallization or by chromatographic methods. The compounds of the formula I according to the invention, especially the compounds without radical of active substance, show sideroforous activity in various strains of gram-negative bacteria. It is therefore possible to use these compounds as a growth factor for certain bacterial cultures. The test on sideroforous activity in accordance with DIN 58 940 was carried out with various bacterial indicator mutants, which do not show growth under the test conditions due to the lack of own iron transport systems. After the administration of the test substances as foreign siderophores a growth promotion can be verified. In the indicator mutants, the biosynthesis of the respective siderophores is blocked, for example pioverdin, pyoqueline, enterobactin, aerobactin, yesiniabactin, or for example aromatic biosynthesis, or the receptors for enterobactin, pyoqueline or pioverdin are missing, as well as other components important for the bacterial transport of iron (for example Cir membrane proteins, Fiu, FepA, and also the TonB protein). Because of this, these mutants can not grow or can only grow very late under conditions of lack of iron. Individually the following indicator mutants were used: Pseudomonas aeruginosa .- PAO 6609, K 407, E. coli: AB 2847, Salmonella typhimurium: enb-7, TA 2700, Yersinia enterocolitica WAH 5030. As controls were used in the case of strains Pseudomonas and Y. enterocolitica, Ferrioxamine E, in the case of the strains Salmonella Ferrioxamina G and Enterobactina, in the case of the E. coli and Ferricromo strains. The test substances were applied in each case in a number of 5μg test sheets. The growth zones of the indicator mutants (diameter in mm) under the influence of the test substances are indicated in Table 1. By virtue of their characteristics as bacterial siderophores, the compounds of the general formula I can serve as a transportation vehicle or penetration accelerators for antimicrobial antibiotics and other active substances, that is to say that conjugated with antibiotics or other active substances, can transport them through iron transport routes to the microbial cell and in this way increase its effectiveness. Accordingly, the compounds of the general formula I with R3, R5, R9 or X = a radical of an active substance, in particular a β-lactam, possess antibacterial activity, partly also with respect to other bacteria resistant to ß -Lactama, being that the sideroforous activity of the benzoxacindion radical is disguised by the antibacterial activity of the complete molecule. In order to determine the antibacterial effectiveness, the minimum inhibition concentrations (MHK) were determined in a microdilution test in accordance with DIN 58 940 in the following bacterial strains: Pseudomonas aeruginosa. : SG 137, NCTC 10662, ATCC 27853, E. coli: ATCC 25922, Kl ebsiella pneumoniae ATCC 10031, Stenotrophomonas mal tophilia GN 12873 and Staphylococcus aureus SG 511. The results of the tests are indicated in table 2. According to it the tested compounds show high antibacterial effectiveness, which in part surpasses that of the comparative substances azlcillin and ampicillin. By varying the iron content of the test agents, and by the use of iron transporting mutants, the dependence of the antibacterial activity of the bacterial transport of iron was demonstrated. The compounds of the general formula I, especially the compounds without radical of active substance, as well as their salts in the case of existing acid groups, and their cleavable esters under physiological conditions, are suitable as siderophores or iron chelators by virtue of of its characteristics to be used as medicines in the case of diseases that derive from a malfunction of the physiological metabolism of iron. The compounds of the general formula I with R3, R5, R9 or X = a radical of an active substance, for example the radical of an antibiotic with an NH or OH group, as well as their salts if there are acid groups, and its splitting esters under conditions • r physiological, are appropriate to be used as drugs against bacterial infections in humans and animals useful by virtue of their antibacterial activity. In the case of the aforementioned diseases, the compounds of the formula I can be used either alone or in the form of pharmaceutical preparations with physiologically acceptable auxiliary substances or carriers known from the state of the art, all of which are possible in principle. the forms of application pharmacological. EXAMPLES Example 1 Preparation of (8-methoxycarbonyloxy-2,4-dioxo-benzoxacin-3-yl) -acetic acid (Formula I with R1 = C00CH3, R2 = H, R3 = CH2COOH) A solution of 2.75 g of glycine in 175 ml of a 0.5M sodium hydrocarbonate solution was cooled in an ultrasound bath at 0-5 ° C. Under stirring, 10.5 g of 2,3-di- (methoxycarbonyloxy) -20-benzoylchloride, dissolved in 20 ml of absolute tetrahydrofuran, were added dropwise at 0-5 ° C. The turbid solution that results after 1 hour is filtered and the tetrahydrofuran is removed by distillation. The substance obtained is separated by suction and washed with a little cold water. For purification the The substance was again dissolved in a 0.5M sodium hydrocarbonate solution, filtered and precipitated with concentrated hydrochloric acid. Colorless crystals with a melting point of 205-208 ° C were obtained in a yield of 70% of the theory. Example 2 Preparation of (8-methoxycarbonyloxy-2,4-dioxo-benzoxazin-3-yl) -acetylglycine (Formula I with R1 = COOCH3, R2 = H, R3 = CH2CONHCH2COOH) The compound was prepared in a manner analogous to Example 1 a from glycylglycine and 2,3-di- (methoxycarbonyloxy) -benzoylchloride in aqueous sodium bicarbonate solution. Colorless crystals with a melting point of 195-198 ° C were obtained after recrystallization in water, in a yield of 70% of the theory. Example 3 Preparation of (8-methoxycarbonyloxy-2,4-dioxo-benzoxacin-3-yl) -acetyl-L-alanine (Formula I with R1 = COOCH3, R2 = H, R3 = CH2CONH-CH (CH3) -COOH) The compound was prepared in a manner analogous to Example 1 from glycyl-L-alanine and 2,3-di- (methoxycarbonyloxy) -benzoylchloride in aqueous sodium bicarbonate solution. Colorless crystals with a melting point of 180-185 ° C were obtained after recrystallization from ethyl acetate, in a yield of 70% of the theory.

Example 4 Preparation of (8-hydroxy-2,4-dioxo-benzoxacin-3-yl acetyl-L-alanine (Formula I with R = H, R H, R '3 CH2CONH-CH (CH3) -COOH) The compound could be obtained by preparative high pressure liquid chromatography (eluent: acetonitrile / water = 1/1 with 0.05% trifluoroacetic acid) from the mother liquor which resulted in isolating the ( 8-methoxycarbonyloxy-2,4-dioxo-benzoxacin-3-yl) -acetyl-L-alanine (product of Example 3). Colorless crystals with a melting point of 203-204 ° C were obtained after recrystallization from acetic acid ethyl ester, in a yield of 20% of the theory. Example 5 Preparation of (8-methoxycarbonyloxy-2,4-dioxo-benzoxacin-3-yl) -acetyl-L-leucine (Formula I with R1 = COOCH3, R2 = H, R3 = CH2CONH-CH (COOH) CH2CH (CH3 ) 2) The compound was prepared in a manner analogous to Example 1 from glycylcholine and 2,3-di- (methoxycarbonyloxy) -benzoylchloride in aqueous sodium bicarbonate solution. The isolation was carried out by immediate extraction with acetic acid ethyl ester of the resulting mixture on acidification with hydrochloric acid, repeated washing with water of the solution thus obtained, drying with sodium sulfate and removal of the solvent in vacuo. After preparative high pressure liquid chromatography (eluent: acetonitrile / water = 1 μl with 0.05% trifluoroacetic acid) and recrystallization from water, colorless crystals with a melting point of 179-181 ° C were obtained in 60% yield of the theory. Example 6 Preparation of (8-hydroxy-2,4-dioxo-benzoxacin-3-yl) -acetyl-L-leucine (Formula I with R1 = H, R2 H, R3 CH2CONH-CH (COOH) CH2CH (CH3) 2) The compound was isolated by preparative high pressure liquid chromatography (eluent: acetonitrile / water = l / l with 0.05% trifluoroacetic acid) as the second product of the reaction mixture obtained during the preparation of (8-methoxycarbonyloxy-2,4-dioxo-benzoxacin-3-yl) -acetyl-L-leucine (product of example 5). After recrystallization from acetic acid ethyl ester, colorless crystals having a melting point of 204-207 ° C were obtained, in a yield of 25% of the theory. Example 7 Preparation of 2-L- (8-methoxycarbonyloxy-2,4-dioxo-benzoxacin-3-yl) -propionic acid (Formula I with R 1 - = COOCH 3, R 2 = H, R 3 = CH (CH 3) -COOH The compound was prepared in a manner analogous to Example 4 from L-alanine and 2,3-di- (methoxycarbonyloxy) -benzoylchloride in aqueous sodium bicarbonate solution. After preparative high pressure liquid chromatography (eluent: acetonitrile / water = 2/3 with 0.05% trifluoroacetic acid) a colorless foam was obtained in a yield of 50% of the theory. Example 8 Preparation of L- (8-methoxycarbonyloxy-2,4-dioxo-benzoxacin-3-ii) -phenylacetic acid (Formula I with R1 = COOCH3, R and « = H, R3 = CAH, -CK-COOH) The compound was prepared analogously to Example 4 from L-phenylalanine and 2,3-di- (methoxycarbonyloxy) -benzoylchloride in aqueous sodium bicarbonate solution. After preparative high pressure liquid chromatography (eluent: acetonitrile / water = l / l with 0.05% trifluoroacetic acid) and recrystallization from water, colorless crystals with a melting point of 182-184 ° C were obtained in a yield of 50% of the theory.

Example 9 Preparation of 4- [(8-methoxycarbonyloxy-2,4-dioxo-benzoxacin-3-ylimino) -methyl-benzoic acid (Formula I with R = COOCH3, R2 = H, R3 = -N = CH-C6H4- COOH (p) To a solution of 0.3 g of 4 - [(2,3-dihydroxybenzoyl) -hydrazonomethyl-benzoic acid in 2 ml of 2N sodium hydroxide and 3 ml of water was added with stirring at 0 ° C 2 ml of ester The mixture was stirred for 30 minutes and then acidified with hydrochloric acid.The crude product was dissolved in hot dimethylformamide, the solution was filtered and the product was separated again by precipitation with water. carried out by preparative high pressure liquid chromatography (eluent: acetonitrile / water = 1 / L with 0.05% trifluoroacetic acid) .Colorless crystals with a melting point of 232-234 ° C. were obtained. EXAMPLE 10 Preparation of N- [ (8-methoxycarbonyloxy-2,4-dioxo-benzoxazin-3-yl) -acetyl-ampicillin (Formula I with R = COOCH3, R2 = H, R3 = CH2CORs, R5 = N-ampicillin-) (a) preparation of (8-methoxycarbonyloxy-2,4-dioxo-benzoxacin-3-yl) -acetylchloride A mixture of 1.07 g of 8-methoxycarbonyloxy -2,4-dioxo-benzoxazin-3-yl-acetic acid (substance 1) and 1 g of phosphorus pentachloride in 5 ml of absolute carbon tetrachloride was heated carefully until the end of HCl development ( 30 minutes) . The resulting solution was filtered and the filtrate was concentrated. The residue was redissolved in hot carbon tetrachloride, the acid chloride was separated by precipitation with anhydrous petroleum ether and dried under high vacuum. Obtained 0.81 g of the product (86% of the theory) with a V " melting point of 80-82 ° C. (b) A solution of 0.78 g of ampicillin sodium salt in 12 ml of 80% tetrahydrofuran was cooled to -5 ° C. To this solution, 0.63 g of 8-methoxycarbonyloxy-2,4-dioxobenzoxazin-3-yl-acetylchloride were added under stirring in 5 portions. The mixture was stirred for 1 hour at 0 ° C and for 1 hour at 20 ° C, and then it was removed by evaporation in vacuo. The residue was mixed with 50 ml of water and 50 ml of acetic acid ethyl ester. Later The mixture was acidified slightly with 1M hydrochloric acid (pH3) and washed three times with an aqueous solution of sodium chloride. The organic phase was separated, dried over sodium sulfate and concentrated to a volume of 20 ml. The residue was mixed with petroleum ether, obtaining 0.92. g (74% of the theory) of the ampicillin derivative as a white powder. Example 11 Preparation of N- [(8-methoxycarbonyloxy-2,4-dioxo-benzoxazin-3-yl) -acetyl-amoxicillin (Formula I with R = COOCH3, R2 = H, R3 = CH2COR5, R5 = N-amoxicillin-) A solution of 0.55 g of amoxicillin in 8 ml of 80% tetrahydrofuran was mixed with 0.22 ml of triethylamine and cooled to -5 ° C. To this solution, 0.45 g of (8-25 methoxycarbonyloxy -2,4-dioxo-benzoxacin-3-yl) -acetylchloride (preparation according to example 8) were added in portions. The mixture was stirred in each case for 1 hour at 0 ° C and at 20 ° C, and then it was removed by evaporation in vacuo. The residue was mixed with 40 ml of water and 40 ml of ethyl acetate. It was acidified slightly with 1M hydrochloric acid (pH3), shaken and washed until the acid was removed with an aqueous solution of sodium chloride. The separated organic phase was dried over sodium sulfate, concentrated to a large extent and mixed with petroleum ether. Thus, the amoxicillin derivative was obtained as a white powder in a yield of 80% of the theory. Example 12 Preparation of N- [(8-methoxycarbonyloxy-2, 4-dioxo-benzoxacin-3-yl) -glycyl-ampicillin (Formula I with R COOCH3, R2 = H, R3 = CH2CONH-CH2-COR5, R5 = N-ampicillin-) 0.352 g of (8-methoxycarbonyloxy-2) , 4-dioxo-benzoxacin-3-yl) -acetyl-glycine (preparation see example 2), 140 μl of triethylamine and a catalytic amount of dimethylaminopyridine were dissolved in 5 ml of absolute tetrahydrofuran, and the solution was mixed under stirring to - 20 ° C with 126 μl isobutyl ester of chloroformic acid. Then 0.357 g of ampicillin sodium salt in 5 ml of 80% tetrahydrofuran was added. It was stirred in each case for 1 hour at -20 ° C and at + 20 ° C, and then it was removed by evaporation in vacuo. The residue was mixed with ethyl acetate and water and carefully acidified with 1M hydrochloric acid. The organic phase was separated, washed three times with sodium chloride solution, dried over sodium sulfate and partially concentrated by evaporation. The ampicillin derivative was precipitated by the addition of petroleum ether, which was purified by preparative high pressure liquid chromatography 'eluent: acetonitrile / water = 1 / L with 0.05% trifluoroacetic acid). Example 13 Preparation of N- [(8-methoxycarbonyloxy-2,4-dioxo-benzoxazin-3-yl) -acetyl-L-alanyl] -ampicillin (Formula I with R1 = COOCH3, R2 = H, R3 = CH2CONH-CH (CH3) -COR5, R5 = N-ampicillin-) The preparation of the compound was carried out analogously to Example 12 starting from (8-methoxycarbonyloxy-2,4-dioxo-benzoxacin-3-yl) -acetyl- L-alanine (preparation see example 3) and sodium salt of ampicillin. Purification was carried out by preparative high pressure liquid chromatography (eluent: acetonitrile / water = 1 / L with 0.05% trifluoroacetic acid), with or that a colorless solid was obtained in a 65% yield of the theory.

Example 14 Preparation of N- [(8-methoxycarbonyloxy -2,4-dioxo-benzoxacin-3-yl) -acetyl-L-leucyl] -ampicillin (Formula I with R1 = COOCH3, R2 = H, R3 = CH2CONH-CH (COR5) CH2CH (CH3) 2, R5 = N-ampicillin-) The preparation of the compound was carried out analogously to Example 12 from (8-methoxycarbonyloxy-2,4-dioxo-benzoxazin-3-yl) -acetyl-L-leucine (preparation see example 5) and sodium salt of ampicillin. Purification was carried out by preparative high pressure liquid chromatography (eluent: acetonitrile / water = 2/3 with 0.05% trifluoroacetic acid), with or a colorless solid was obtained in a 60% yield of the theory. Example 15 Preparation of N- [(8-methoxycarbonyloxy-2,4-dioxo-benzoxacin-3-yl) -acetyl] -0-n-propionyl-amoxicillin (Formula I with R1 = C00CH3, R2 = H, R3 = CH2C0R5 , Rs = N- (0-n-propionyl) -amoxicilino-) 0.385 g of N- [(8-methoxycarbonyloxy-2,4-dioxo-benzoxacin-3-yl) -acetyl-amoxicillin (preparation see example 11) dissolved in 25 ml of tetrahydrofuran and the solution was cooled to -78 ° C. Under stirring, 0.34 ml of triethylamine was added, followed by 0.16 ml of propionyl chloride. The reaction mixture was stirred for 30 minutes at -60 ° C and for 1 hour at 20 ° C. It was then evaporated off under vacuum and the residue was mixed with water and ethyl acetate. After acidification with 1M hydrochloric acid (pH3), it was washed until the acid was removed with an aqueous solution of sodium chloride. The separated organic phase was dried over sodium sulfate, concentrated to a large extent and mixed with petroleum ether. With this the amoxilin derivative was obtained as a white powder in a yield of 40% of the theory. Example 16 Preparation of 4- (8-methoxycarbonyloxy-2,4-dioxo-benzoxacin-3-yl) -benzoic acid (Formula I with R1 = C00CH3, R2 = H, R3 = C6H4-COOH (p) The compound was prepared Analogously to Example 1 from 4-aminobenzoic acid and 2,3-di- (methoxycarbonyloxy) -benzoylchloride in aqueous sodium hydrocarbonate solution After recrystallization from acetic acid ethyl ester, colorless crystals were obtained with a fusion of 236-240 ° C, in a yield of 80% of the theory Example 17 Preparation of 2L, 6-bis (8-methoxycarbonyloxy-2, -dioxo-1,3-benzoxacin-3-yl) -hexanic acid (Formula I with R1 C00CH3, R2 = H, R3 = CHR4-C00H, R4 = (CH2) 4Y) The compound was obtained as a colorless foam in a manner analogous to Example 5, from L-lysine and 2 molar equivalents of , 3-di- (methoxycarbonyloxy) -benzoylchloride in aqueous sodium hydrocarbonate solution Example 18 Preparation of sodium salt of N- [4- (8-methoxycarbonyloxy-2, 4-d ioxo-1,3-benzoxacin-3-yl) -benzoylj-ampicillin (Formula I with R1 = COOCH3, R2 = H, R3 = R20 with R13 = H, COR9 in position 4, R = (sodium salt) of N -ampicillin), p = 0) (a) Preparation of 4- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxacin-3-yl) -benzoylchloride The compound was prepared analogously to the substance 10a a from substance 16 and phosphorus pentachloride in carbon tetrachloride. A colorless oil was obtained in a 65% yield of the theory. (b) preparation of N- [4- (8-methoxycarbonyloxy-2,4-dioxo-l, 3-benzoxacin-3-yl) -benzoyl] -mpieylin The compound was prepared analogously to substance 10b from 4- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) -benzoylchloride and sodium salt of ampicillin. A white powder was obtained in a yield of 85% of the theory. (c) sodium salt: A solution of 0.25 g of N- [4- (8-methoxycarbonyloxy-2,4-dioxo-l, 3-benzoxacin-3-yl) -benzoyl] -ampicillin in acetic acid ethyl ester It is mixed under cooling on ice and stirring with a solution J 0.083 g of sodium 2-ethylhexanoate in ethyl acetate. After extracting the product by precipitation with petroleum ether, it was separated by filtration. Purification was carried out by preparative high pressure liquid chromatography (eluent: acetonitrile / water). With this a white powder was obtained in a yield of 0.202 g (79% of the theory). Example 19 Preparation of 4- (8-methoxycarbonyloxy-2) acid, 4-dioxo-l, 3- 1 2-benzoxacin-3-yl) -benzoic acid (Formula I with R = COOCH3, R = H, R3 = R2 ° with R13 = * H, COR9 in position 4, R9 = OH , p = 1) The compound was prepared analogously to substance 5 from 4- (aminomethyl) -benzoic acid and 2,3-di- (methoxycarbonyloxy) -benzoylchloride in solution aqueous sodium hydrocarbonate. After recrystallization from acetic acid ethyl ester, colorless crystals with a melting point of 220-222 ° C were obtained, in a yield of 65% of the theory. Example 20 Preparation of sodium salt of N- [4- (8-methoxycarbonyloxy- 2,4-dioxo-l, 3-benzoxacin-3-methyl-methyl) -benzoyl] -ampieiline (Formula I with R1 = COOCH3, R2 = H, R3 = R20 with R13 = H, COR9 in position 4, R = (sodium salt) of N-ampicillin), p = 1) (a) preparation of 4- (8-methoxycarbonyloxy -2, 4- dioxo- 25 1, 3-benzoxacin-3-methyl-methyl) -benzoylchloride • ^ The compound was prepared analogously to substance 10a from substance 19 and phosphorus pentachloride in carbon tetrachloride. A colorless oil was obtained in a 95% yield of theory. 5 (b) preparation of N- [4- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-methyl-methyl) -benzoyl] -mpieylin The compound was prepared analogously to substance 10b from 4- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-methyl-methyl) -benzoylchloride and sodium salt of ampicillin. A white powder was obtained in a yield of 80% of the theory. (c) sodium salt: the preparation was carried out analogously to substance 18 from N- [4- (8-methoxycarbonyloxy -2,4-dioxo-l, 3-benzoxacin-3-methyl-methyl) ) - 15 benzoylj-ampicillin and 2-ethylhexanoate sodium in acetic acid ethyl ester. A white powder was obtained in a yield of 60% of the theory. Example 21 Preparation of 3,5-bis- (8-methoxycarbonyloxy-2,4,4-dioxo-1,3-benzoxazin-3-yl) -benzoic acid (Formula I with R C00CH3, R2 = H, R3 = R20 with R13 = Y in position 3, COR9 in position 5, R9 = OH, p = 0, R1, R2 as above) The compound was prepared analogously to substance 5 from 3,5-diaminobenzoic acid and molar equivalents of 2,3-di- (methoxycarbonyloxy) -benzoylchloride in aqueous sodium hydrocarbonate solution. After recrystallization from ethyl acetate, colorless crystals having a melting point of 164-166 ° C were obtained, in a yield of 65% of the theory. EXAMPLE 22 Preparation of sodium salt of N- [3,5-bis- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxacin-3-yl) -benzoyl] -ampicillin (Formula I with R 1 = COOCH 3 , R2 = H, R3 = R20 with R-Y in position 3, COR in position 5, R = (sodium salt) of N-ampicillin), p = 0, R1, R as above) (a) preparation of 3 , 5-bis- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) -benzoylchloride The compound was prepared analogously to substance 10a from substance 21 and phosphorus pentachloride in carbon tetrachloride. It was obtained in a 90% yield of the theory as colorless foam. (b) preparation of N- [3,5-bis- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxacin-3-yl) -benzoyl] -mpieylin The compound was prepared analogously to the substance 10b from 3, 5-bis- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxacin-3-yl) -benzoylchloride and sodium salt of ampicillin. It was obtained as white powder in a yield of 80% of the theory. (c) sodium salt: the compound was prepared in a manner and Analogous to substance 18 from N- [3,5-bis- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) -benzoyl] -amicillin and sodium 2-ethylhexanoate in ethyl ester of acetic acid. A white powder was obtained in a yield of 40% of the theory. Example 23 Preparation of 3- (8-methoxycarbonyloxy-2,4-dioxo-l, 3-benzoxacin-3-yl) -propionic acid (Formula I with R1 = COOCH3, R2 = H, R3 = (CH2) 2COOH) The compound was prepared analogously to substance 5 from β-alanine and 2,3-di- (methoxycarbonyloxy) -benzoylchloride in aqueous sodium hydrocarbonate solution. After recrystallization from acetic acid ethyl ester crystals were obtained colorless with a melting point of 140-144 ° C, in a yield of 55% of the theory. Example 24 Preparation of sodium salt of N- [3- (8-methoxycarbonyloxy- 2,4-dioxo-l, 3-benzoxacin-3-yl) -propionyl] -ampicillin 20 (Formula I with R1 = C00CH3, R2 = H, R3 = (CH2) 2C0-N-ampicillin (sodium salt) (a) preparation of 3- (8-methoxycarbonyloxy-2,4-dioxo-1, -benzoxacin-3-yl) -propionylchloride The compound was prepared analogously to the Substance 10a from substance 23 and phosphorus pentachloride in carbon tetrachloride. A yellowish oil was obtained in a 100% yield of the theory. (b) preparation of N- [3 - (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) -propionyl] -ampicillin The compound was prepared analogously to substance 10b from 2- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) -propionylchloride and sodium salt of ampicillin. A white powder was obtained in a yield of 88% of the theory. (c) sodium salt: the salt was prepared analogously to substance 18 from N- [3- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) -propionylj- ampicillin and sodium 2-ethylhexanoate in ethyl acetate. A white powder was obtained in a yield of 41% of the theory. Example 25 Preparation of 3,5-bis - [3 - (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) -propionylamino] -benzoic acid (Formula I with R1 = C00CH3, R2 = H, R3 = R18 with R10, R11 = H, R12 = R19 in position 3, COR9 in position 5, R9 = OH, R15, R14 = H, n = 2, R1, R as above) The compound was prepared in a analogous to substance 5 from 3,5-diaminobenzoic acid and 2 molar equivalents of 3- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) -propionylchloride (substance 24a) in solution aqueous sodium hydrocarbonate. Colorless crystals with a melting point of 160-165 ° C were obtained, in a yield of 50% of the theory. Example 26 Preparation of sodium salt of N- [3,5-bis- [3- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) -propionylamino] -benzoyl] -ampicillin ( Formula I with R = C00CH3, R2 = H, R3 = R18 with R10, R11 = H, R12 = R19 in position 3, COR in position 5, R = N-ampicillin (sodium salt), R15, R14 = H, n = 2, R1, R2 as above) (a) preparation of N- [3, 5-bis- [3- (8-methoxycarbonyloxy-2,4-dioxo-l, 3-benzoxazin-3-yl) -propionylamino ] -benzoi1] -ampicillin The compound was prepared analogously to substance 12 from substance 23 and sodium salt of ampicillin. It was obtained as white powder in a yield of 80% of the theory. (b) sodium salt: the compound was prepared in a manner analogous to substance 18 from N- [3,5-bis- [3- (8-methoxycarbonyloxy) -2,4-dioxo-1,3-benzoxacin- 3-yl) propionylamino] -benzoyl] -ampicillin and sodium 2-ethylhexanoate in ethyl acetate. A white powder was obtained in a yield of 18% of the theory.

Example 27 Preparation of 3,5-bis- [(8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxacin-3-yl) -acetylamino] -benzoic acid (Formula I with R 1 = COOCH 3, R 2 = H, R3 = R18 with R10, R11 = H, R12 = R19 in position 3, COR9 in position 5, R9 = OH, R15, R14 = H, n = 1, R1, R2 as above) The compound was prepared analogously to substance 5 from 3,5-diaminobenzoic acid and 2 molar equivalents of (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) -acetylchloride (substance 10a) in aqueous hydrocarbonate solution of sodium. After recrystallization from ethyl acetate, colorless crystals with a melting point of 190-195 ° C were obtained in a yield of 53% of the theory. EXAMPLE 28 Preparation of sodium salt of N- [3,5-bis- [(8-methoxycarbonyloxy-2,4-dioxo-l, 3-benzoxacin-3-yl) -acetylamino] -benzoyl] -ampicillin (Formula I with R1 = C00CH3, Ri-, 2 = THT, iR-, 3 = tR-, 8 with nR10, tR-, 11 = tHt, tR-, 12 = _R19 in posi.ci.ó ^ n 3_., COR 9 in position 5, R9 N-ampicillin (sodium salt), R15, R14 = H, n = 1, R1, R2 as above) (a) preparation of N- [3,5-bis- [(8 - methoxycarbonyloxy-2,4-dioxo-l, 3-benzoxacin-3-yl) -acetylamino] -benzoyl] -ampicillin The compound was prepared analogously to substance 12 from substance 27 and sodium salt of ^ ampicillin It was obtained as a white powder in a yield of 90% of the theory. (b) sodium salt: the salt was prepared analogously to substance 18 from N- [3, 5-bis- [(8,5-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxacin-3-yl) -acetylamino] -benzoii] -ampicillin and sodium 2-ethylhexanoate in ethyl acetate. A white powder was obtained in a yield of 17% of the theory. Example 29 Preparation of 4-chloro-3- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) -benzoic acid (Formula I with R = C00CH3, R2 = H, R3 = R20 with R13 = Cl in position 2, COR9 in position 5, R = OH, p = 0) The compound was prepared analogously to substance 5 from -amino-4-chloro-benzoic acid and 2, 3 - di- (methoxycarbonyloxy) -benzoylchloride in aqueous sodium hydrocarbonate solution. After recrystallization from ethyl acetate, colorless crystals with a melting point of 20 234-236 ° C were obtained, in a yield of 41% of the theory. Example 30 Preparation of sodium salt of N- [4-chloro-3- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxacin-3-yl) -benzoyl] -amicillin (Formula I with R1 = COOCH3 , R2 = H, R3 = R 20 with R = Cl in position 2, COR in position 5, R = N-ampicillin (sodium salt), p = 0) (a) preparation of 4-chloro-3- (8-methoxycarbonyloxy-2,4-dioxo- 1, 3-benzoxacin-3-yl-) -benzoylchloride The compound was prepared analogously to substance 10a from substance 29 and phosphorus pentachloride in carbon tetrachloride. A yellowish powder was obtained in a yield of 94% of the theory with a melting point of 76-78 ° C. (b) preparation of N- [4-chloro-3- (8-methoxycarbonyloxy-2,4-dioxo-l, 3-benzoxazin-3-yl) -benzoyl] -mpieylin The compound was prepared analogously to the substance 10b from 4-chloro-3- (8-methoxycarbonyloxy- 2,4-dioxo-l, 3-benzoxacin-3-yl) -benzoylchloride and sodium salt of ampicillin. It was obtained as a white powder in a yield of 87% of the theory. (c) sodium salt: prepared in a manner analogous to substance 18 from N- [4-chloro-3- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) - benzoyl] -ampicillin and sodium 2-ethylhexanoate in ethyl acetate. A white powder was obtained in a 55% yield of theory. Example 31 Preparation of 2-hydroxy-4- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxacin-3-yl) -benzoic acid (Formula I with R C00CH3, R2 = H, R3 = R20 with R13 = OH in position 3, COR9 in position 4, R9 = OH, p = 0) The compound was prepared analogously to substance 5 from 4-aminosalicylic acid and 2,3-di- (methoxycarbonyloxy) -benzoylchloride in aqueous solution of sodium hydrocarbonate. After recrystallization from acetic acid ethyl ester, colorless crystals with a melting point of 261-262 ° C were obtained, in a yield of 68% of the theory. Example 32 Preparation of sodium salt of N- [2-hydroxy-4- (8-methoxycarbonyloxy-2,4-dioxo-l, 3-benzoxacin-3-yl) -benzoyl] -ampicillin (Formula I with R1 = C00CH3 , R2 = H, R3 = R20 with R1 = OH in position 3, COR in position 4, R9 = N-ampicillin (sodium salt), p = 0) (a) preparation of succinimido-2-hydroxy-4- ( 8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxacin-3-yl-) -benzoate: To a solution of 0.224 g of substance 31 and 0.069 g of N-hydroxysuccinimide in 5 ml of anhydrous dioxane was added. under stirring at 0 ° C, a solution of 0.124 g of dicyclohexylcarbodimide in 5 ml of anhydrous dioxane. The mixture was stirred for 8 hours at 20 ° C, the formed precipitate was removed by filtration, and the solvent was removed in vacuo. The remaining oil was solidified by trituration with some isopropanol. Recrystallization from acetic acid ethyl ester gave a white powder in a yield of 0.23 g (81% of theory) with a melting point of 145-150 ° C. (b) Preparation of N- [2-hydroxy-4- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) -benzoyl] -mpieylin: Under argon atmosphere 0.191 g of ampicillin trihydrate in a mixture of 5 ml of tetrahydrofuran and 5 ml of water, and brought to solution with 138 μl of triethylamine. A solution of 0.223 g of succinimido-2-hydroxy-4- (8-methoxycarbonyloxy-2,4-dioxo-l, 3-benzoxacin-3-yl) -benzoate in 5 ml of tetrahydrofuran was then added under stirring, and the The mixture was stirred for 10 hours at 20 ° C. The reaction mixture was concentrated by evaporation at 20 ° C, and the residue was mixed with water and ethyl acetate. After acidification, the organic phase was separated, washed with a sodium chloride solution and with water, dried over sodium sulfate and finally concentrated largely by evaporation. By the addition of petroleum ether, the ampicillin derivative was precipitated, which was obtained with a yield of 0.28 g (84% of the theory). (c) sodium salt: the salt was prepared analogously to substance 18 from N- [2-hydroxy-4- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) ) -benzoyl] -ampicillin and sodium 2-ethylhexanoate in ethyl acetate. A white powder was obtained in a yield and of 19% of the theory. Example 33 Preparation of 3- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) -benzoic acid (Formula I with R1 = COOCH3, R2 = 5 H, R3 = R20 with R * 3 = H, COR9 in position 3, R9 = OH, p = 0) The compound was prepared in a manner analogous to substance 5 from 3-aminobenzoic acid and 2,3-di- (methoxycarbonyloxy-benzoylchloride in aqueous sodium hydrocarbonate After recrystallization from acetic acid ethyl ester, colorless crystals were obtained in a yield of 71% of the theory with a melting point of 250-253 ° C. EXAMPLE 34 Preparation of sodium salt of N- [3- (8-methoxycarbonyloxy-2, 4-dioxo-l, 3-benzoxacin-3-yl) -benzoyl] -ampicillin (Formula I with R1 = COOCH ,, R2 = H, R3 = R20 with R13 = H , COR9 in position 3, R = N-ampicillin (sodium salt), p = 0) (a) preparation of 3- (8-methoxycarbonyloxy -2,4-dioxo-1,3-benzoxacin-3-yl) -benzoylchloride 20 The compound was prepared analogously to the substance 10a from substance 33 and phosphorus pentachloride in carbon tetrachloride. A yellowish oil was obtained in a yield of 97% of the theory. (b) preparation of N- [3- (8-methoxycarbonyloxy-2,4,4-dioxo-1,3-benzoxazin-3-yl) -benzoyl] -ampicillin and The compound was prepared analogously to substance 10b from 3- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxacin-3-yl) -benzoylchloride and sodium salt of ampicillin. A white powder was obtained in a yield of 87% of the theory. (c) sodium salt: the salt was prepared analogously to substance 18 from N- [3 - (8-methoxycarbonyloxy- 2,4-dioxo-1,3-benzoxacin-3-yl) -benzoylj- ampicillin and sodium 2-ethylhexanoate in ethyl acetate.

A white powder was obtained in a yield of 59% of the theory. EXAMPLE 35 Preparation of 2L- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) pentane-dicarboxylic acid benzyl ester (Formula I with R1 = COOCH3, R2 = H, R3 = CHR4 - COR5 with R4 = (CH2) 2COOH, R5 = O-benzyl) 1 g of 1-glutamic acid benzyl ester were dissolved under an argon atmosphere in 40 ml of anhydrous tetrahydrofuran. Under cooling on ice and After stirring, 1.24 ml of triethylamine was added, followed by a solution of 1.22 g of 2,3-di- (methoxycarbonyloxy) -benzoylchloride in 10 ml of anhydrous tetrahydrofuran. After stirring for 20 hours at 20 ° C, the tetrahydrofuran was removed in vacuo and the The residue was mixed with water and ethyl acetate. While cooling on ice and stirring it was acidified, and then the ethyl ester phase of acetic acid was separated. It was washed several times with water and a saturated solution of sodium chloride, and finally it was concentrated by evaporation. The purification was carried out by preparative high pressure liquid chromatography. A yellowish foam was obtained in a yield of 0.260 g (13% of theory). Example 36 Preparation of the sodium salt of the 4-ampicillincarbamoyl-2L- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxacin-3-yl) -butyric acid benzyl ester (Formula I with R - = COOCH3 , R2 = H, R3 = CHR4-COR5 with R4 = (CH2) 2CO-N-ampicillin (sodium salt), R5 = 0-benzyl) (a) preparation of benzyl ester of 4-chlorocarbonyl-2L- (8) -methoxycarbonyloxy-2,4-dioxo-1,3-benzoxacin-3-yl-) -butyric acid The compound was prepared analogously to substance 10a from substance 35 and phosphorus pentachloride in carbon tetrachloride. A yellowish oil was obtained in a yield of 97% of the theory. (b) preparation of 4-ampicillincarbamoyl-2L- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxacin-3-yl) -butyric acid benzyl ester The compound was prepared analogously to substance 10b a from 4-chlorocarbonyl-2L- (8-methoxy-carbonyloxy-2,4-dioxo-1,3-benzoxacin-3-yl) -butyric acid and sodium salt of ampicillin. A white powder was obtained in a yield of 87% of the theory. (c) sodium salt: prepared analogously to substance 18 from 4-ampicillincarbamoyl-2L- (8-methoxycarbonyloxy-2), 4-dioxo-l, 3-benzoxacin-3-yl) -butanoyl) -ampicillin and sodium 2-ethylhexanoate in ethyl acetate. A white powder was obtained in a 46% yield of theory. Example 37 Preparation of 2L- [2L, 6-bis- (8-methoxycarbonyloxy-2,4-dioxo-l, 3-benzoxacin-3-yl) -hexanoylamino] -6- (8-methoxy-carbonyloxy-2, 4-dioxo-l, 3-benzoxacin-3-yl) -hexanic (Formula I with R1 = * COOCH3, R2 = H, R3 = CHR4 - COR5 with R4 = (CH2) 4-Y, R5 = R17 with X = OH, n = 4) (a) Preparation of the benzylic ester of 2L- [2L, 6-bis- (8-methoxycarbonyloxy-2,4-dioxo-l, 3-benzoxacin-3-yl) -hexanoylamino] -6 - (8-methoxy-carbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) -hexanic: 1.55 g of tris-p-toluenesulfonate of the benzylic ester of 6-amino-2- (2, 6) -diaminohexanoylamino) -hexanic were dissolved under an argon atmosphere in 20 ml of anhydrous dimethylformamide and, under cooling on ice and stirring, the solution was mixed first with 1.48 ml of triethylamine and then with a solution of 1.53 g of 2,3-di- (methoxycarbonyloxy) -benzoylchloride in 10 ml of anhydrous dimethylformamide. After stirring for 5 hours at 0 ° C and for 20 hours at 20 ° C, the dimethylformamide was removed in vacuo and the residue was mixed with water and ethyl acetate. While cooling on ice and stirring it was acidified, and then the ethyl ester phase of acetic acid was separated. It was washed several times with water and a saturated solution of sodium chloride, and finally it was concentrated by evaporation. Purification was carried out by preparative high pressure liquid chromatography (eluent: acetonitrile / water = 1 / L with 0.05% trifluoroacetic acid), leaving 0.25 g of a yellowish oil (14% of the theory). (b) 0.250 g of benzylic acid ester of 2L- [2L, 6-bis- (8-methoxycarbonyloxy -2,4-dioxo-1,3-benzoxacin-3-yl) -hexanoylamino] -6- (8-methoxy) -carbonyloxy-2,4-dioxo-l, 3-benzoxacin-3-yl) -hexanic acid were catalytically hydrated at 20 ° C and normal pressure in 30 ml of ethanol on 0.06 g of palladium on activated carbon (10% of Pd) . After filtering over celite it was concentrated by evaporation and solidified with ethyl acetate. A yellowish foam was obtained in a yield of 0.220 g (98% of theory). Example 38 Preparation of the sodium salt of N- [2 L- [2 L, 6-bis- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) -hexanoyl-amino] -6- (8-methoxy-carbonyloxy-2,4-dioxo-l, 3-benzoxacin-3-yl) -hexanoyl] -ampicillin (Formula I with R1 = COOCH3, R2 = H, R3 = CHR4-COR5 with R4 = (CH2 ) 4-Y, R5 = R17 with X = N-ampicillin (sodium salt), n = 4) (a) preparation of N- [2L- [2L, 6-bis- (8-methoxy-carbonyloxy-2) , 4-dioxo-l, 3-benzoxacin-3-yl) -hexanoyl-amino] -6- (8-methoxy-carbonyloxy-2,4-dioxo-l, 3-benzoxacin-3-yl) -hexanoyl] - ampicillin The compound was prepared analogously to substance 12 from substance 37 and sodium salt of ampicillin. A white powder was obtained in a yield of 90% of the theory. (b) sodium salt: the salt was prepared in a manner analogous to substance 18 from N- [2L- [2L, 6-bis- (8-methoxy-carbonyloxy-2,4-dioxo-1,3- benzoxacin-3-yl) -hexanoyl-amino] -6- (8-methoxy-carbonyloxy-2,4-dioxo-l, 3-benzoxacin-3-yl) -hexanoyl] -ampicillin and sodium 2-ethylhexanoate in ester Acetic acid ethyl ester. A white powder was obtained in a yield of 15% of the theory. Example 39 Preparation of 3-hydroxy-2L- (8-methoxycarbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) -propaneic acid (Formula I with R = COOCH3, R2 = H, R3 = CHR4 - COR5 with R4 = CH2-OH, R5 = OH) (a) preparation of the benzylic ester of 2L-N- (2,3-dimethoxycarbonyloxybenzoyl) -serine The compound was prepared analogously to the substance 35 from the ester hydrochloride benzyl of L-serine and 2,3-di-methoxycarbonyloxy) -benzoylchloride in tetrahydrofuran. White foam was obtained in a 43% yield of theory. (b) 1.3 g of the benzyl ester of 2L-N- (2) was suspended, 3-dimethoxycarbonyloxybenzoyl) -serine in 50 ml of ethanol and 300 mg of palladium on carbon catalyst (10%) were added. After transfer to a hydrogen atmosphere, the mixture was shaken for 2 hours at 20 ° C and normal pressure. After filtration over celite it was concentrated by evaporation. Purification by preparative high pressure liquid chromatography resulted in a white foam, yield: 599 mg (48% of theory). Example 40 Preparation of the sodium salt of N- [3-hydroxy-2L- (8-methoxycarbonyloxy -2,4-dioxo-l, 3-benzoxacin-3-yl) -1,3-propionyl] -ampicillin (Formula I with R = COOCH3, R = H, R = CHR4-COR5 with R4 = CH2-OH, R5 = N-ampicillin (sodium salt)) (a) preparation of the N- [3-hydroxy-2L- (8- methoxycarbonyloxy -2,4-dioxo-l, 3-benzoxacin-3-yl) -propionyl] -ampicillin The compound was prepared analogously to substance 12 from substance 39 and sodium salt of ampicillin. A white powder was obtained in a yield of 90% of the theory. (b) sodium salt: prepared in a manner analogous to substance 18 from N- [3-hydroxy-2L- (8-methoxy-carbonyloxy-2,4-dioxo-1,3-benzoxazin-3-yl) ) -propionyl] -ampicillin and sodium 2-ethylhexanoate in ethyl acetate. A white powder was obtained in a yield of 10% of the theory.

Example 41 Preparation of the acid (8-ethoxycarbonyloxy-2,4-dioxo-1-benzoxacin-3-yl) -acetic acid (Formula I with R = COOC2H5, RH, R3 = CH2COOH) The compound was prepared in a manner analogous to Example 1 a from glycine and 2,3-di- (ethoxycarbonyloxy) -benzoylchloride in aqueous sodium hydrocarbonate solution. After recrystallization from acetic acid ethyl ester, colorless crystals with a melting point of 162-163 ° C were obtained, in a yield of 69% of the theory.

Example 42 Preparation of N- [N '- [6 - (8-methoxycarbonyloxy-2,4-dioxo-benzoxacin-3-yl) -hexyl] -N' - [2,3-di- (methoxycarbonyloxy) -benzoyl] 6-aminohexyl] -N- [2,3-di- (methoxycarbonyloxy) -benzoyl] -glycine (Formula I with R3 = Z-CHR4-COR5; R1, R7 = COOCH3, R2, R4, RS = H, R5 = OH, n = 6, m = 2) (a) Benzyl ester preparation (R5 = OCH2C6H5): To a solution of 880 mg of benzylic ester tosylate N- [N '- (6-aminohexyl) -6- aminohexyl] -glycine and 1.04 ml of triethylamine in 20 ml of dichloromethane was added at -30 ° C a solution of 864 mg of 2,3-di- (methoxycarbonyloxy) -benzoylchloride in 5 ml of dichloromethane. The mixture was stirred for 1 hour at -10 ° C and for 1 hour at room temperature, and then filtered. The solution was concentrated and the residue was taken up in 20 ml of acetic acid ethyl ester. The acetic acid ethyl ester solution was washed three times with 1 M hydrochloric acid, saturated sodium carbonate solution and with water. After drying and removing the solvent by vacuum distillation, a colorless solid was obtained in a yield of 650 mg (60% of the theory) (b) acid preparation ((R = OH) The preceding benzyl ester was hydrated in methanol with 100 mg Pd / C (10%) for 3 hours in a shaker at room temperature After removing the catalyst by filtration over celite, the solvent was removed by vacuum distillation. a 70% performance of the theory.

Table 1: Growth zones (in mm) of siderophore indicator strains with selected benzoxazidene derivatives a) Ferrioxamine E, b) Ferricrome, c) Ferrioxamine G c Table 2: Antibacterial activity of the benzoxationdione-antibiotic conjugates MHK values in mg / ml V In the examples HPLC means high pressure liquid chromatography

Claims (22)

1. r CLAIMS Benzoxazidene Derivatives of Formula I wherein R1 = H, COalkyl or COOalkyl, R2 = H, Alkyl, halogen, and R represents the following sust:? T: u; yent is: R3 = Z - - CHR 4 - COR5 with Z _ twenty with R 4 = H, alkyl, phenyl or substituted phenyl, especially hydroxyphenyl or acyloxyphenyl, or with R 4 = (CH 2) nCOX with X = OA, where A = H, Alkyl, an alkali metal ion or an ammonium ion, is either a substituted ammonium ion, or with X = a radical of active substance, in epsecial a radical of an antibiotic, which is linked through a group OH or an NH group, and with n = 1 -10, or with R = (CH2) nY, where Y represents a radical of bezoxane dione of the form wherein R1 and R2 = are as defined above, and both bezoxacezone radicals may be the same or different, and may be n = 1 -10, or with Rs = OA, where A is defined as above, or with R5 = a radical of active substance, in particular a radical of an antibiotic, which is linked through an OH group or an NH group, or with R5 = NH-CHR8-COR9, with R8 = H, alkyl, phenyl or substituted phenyl and with R9 = OA, where A is defined as above, or with R = a radical of active substance, especially a radical of an antibiotic, which is linked through an OH group or an NH group, or with R ~ = 17 = R with X and Y as above, n = 1 10 and with R = H, alkyl, halogen and with R = H, acyl and n = 1-10 and m = 1 2, or b) R3 = CHR4 -COR5 with R and R as above, or 18 = R with R1 and / or RH, alkyl, phenyl or substituted phenyl, n = 1-10, and with COR9 and R12 in all possible positions, R defined as above and R12 = H, alkyl, halogen, hydroxy, alkoxy , a radical Y of benzoxacose, or = R * with R1, R2 as above, R14, R15 as R1, R2 and can be n = 1 10, d) RJ = = R20 with R and COR in all possible positions, and with 13 R = H, alkyl, halogen, hydroxy, alkoxy or a Y radical of benzoxacezione, and R as above and p = 0-2, or e) R3 = R3 = = R22 with R as above, R = H, alkyl, phenyl or substituted phenyl, of) R - = to a radical of active substance, especially a radical of an antibiotic, which is linked through an OH group or a NH group, as well as the cleavable esters under physiological conditions of those compounds of the formula I having a free carboxyl group in the radical R3.
2. 2. Compounds of the formula I according to claim 1, wherein R1 = carboxyalkyl, R2 = H, R3 = CHR4-COR5, R4 = H, alkyl, phenyl or p-hydroxyphenyl, and R = OH.
3. 3. Compounds of the formula I according to claim 1, wherein R = carboxyalkyl, R2 = H, R3 = CH2CONHCHR8, with R8 = H, alkyl, phenyl or substituted phenyl.
4. 4. (8-methoxycarbonyloxy-2,4,4-dioxo-benzoxacin-3-yl) -acetic acid according to claim 1.
5. 5. (8-methoxycarbonyloxy-2,4-dioxo-benzoxacin-3-yl) -acetylglycine according to claim 1.
6. 6. Compounds of the formula I according to claim 1, wherein X or R or R or R means the radical of an antibacterial active substance.
7. 7. Compounds of the formula I according to claim 1, wherein X or R or R or R means the radical of any cephalosporin that is linked through an NH or OH group.
8. 8. Compounds of the formula I according to claim 1, wherein X or R or R5 or R9 means »b the radical of any penicillin that is linked through an NH or OH group.
9. 9. Compounds of the formula I according to claim 1 or 8, wherein X or R3 or R5 or R9 5 means a radical of ampicillin.
10. 10. Compounds of the formula I according to claim 1 or 8, wherein X or R3 or R5 or R9 means a radical of amoxycillin or of O-acylamoxycillin.
11. 11. Compounds of the formula I according to claim 1, wherein X or R or R5 or R9 means a radical of 6-aminopenicilane acid.
12. 12. Compounds of the formula I according to claim 1, wherein X or R or R5 or R9 means 15 any tetracycline radical that is linked through an NH or OH group.
13. 13. Compounds of the formula I according to claim 1, wherein X or R3 or R5 or R9 means the radical of any macrolide that is linked 20 through a NH or OH group.
14. 14. Compounds of the formula I according to claim 1, wherein X or R or R or R means the radical of any quinolone that is linked through an NH or OH group.
15. 15. Compounds of the formula I according to claim 3, wherein X or R or R or R means the radical of any carbapenem which is linked through an NH or OH group.
16. 16. N- (8-methoxycarbonyloxy-2,4-dioxo-benzoxacin-3-yl-acetyl) -ampicillin according to claim 1, 8 or 9.
17. 17. N- (8-methoxycarbonyloxy-2,4-dioxo) -benzoxacin-3-yl-acetyl) -amoxiciline according to claim 1, 8 or 10.
18. 18. Use of the compounds of the formula I according to one of claims 1 and 17 as growth factors for bacteria.
19. Use of the compounds of the formula I according to one of claims 1 to 17 which contain a radical of active substance for X or R or R or R as therapeutic agents in the case of bacterial infections.
20. 20. Use of the compounds of the formula I according to one of claims 1 to 17 as therapeutic agents in the case of diseases resulting from a disorder in the metabolism of iron.
21. 21. Medicament containing a compound of formula I according to one of claims 1 to 17, together with the usual carrier materials.
22. 22. Compounds of the formula I, according to one of claims 1 to 17, for use as a medicament. SUMMARY The invention relates to new benzoxazioner derivatives of the general formula I, and to their conjugates with active substances, for example antibiotics. The compounds according to the invention represent protected catechol derivatives in heterocyclic form and are effective as siderophores in the case of strains of gram-negative bacteria, especially pseudomonates, E. coli and salmonella strains. In the form of their conjugates with active substances, for example antibiotics (such as "siderophore-antibiotic conjugates"), they can introduce these into bacterial cells and improve or extend their antibacterial effectiveness, partly also with respect to other bacterial strains resistant to β -Lactama Furthermore, the aforementioned compounds are suitable as potential prodrug factors for iron chelators, for use in the case of diseases resulting from iron metabolism. The application of the invention can be carried out in pharmaceutical research, industry and agronomy. Formula I wherein R 1 = H or carboxyalkyl, R 2 = H, alkyl or phenyl, and R various acid groups derived from amino acids, dipeptides and hydrazones, or their conjugates with active substances, for example antibiotics. The indicated compounds can be present as free acids, in the form of their salts or as easily unfoldable steres.