EP0923538A1

EP0923538A1 - Compounds which can form complexes with metals

Info

Publication number: EP0923538A1
Application number: EP96923943A
Authority: EP
Inventors: Wolfram Trowitzsch-Kienast; Rolf Reissbrodt; Horst-Dieter Ambrosi; Vera Hartmann
Original assignee: Analyticon AG Biotechnologie Pharmazie
Current assignee: Analyticon AG Biotechnologie Pharmazie
Priority date: 1994-12-22
Filing date: 1996-06-26
Publication date: 1999-06-23
Also published as: WO1997049669A1; DE4447374A1; DE19626020A1; AU6417296A

Abstract

Compound of general formula (I) where R = -CH2-NH-CO-(2,3-dihydroxyphenyl), CN or -CH2-NH2, n = 1 to 5.

Description

Compounds that can form complexes with metals

The present invention relates to a compound of the general formula according to claim 1, an iron complex of this compound according to claim 3, a process for the preparation of the compounds according to claim 4, a conjugate according to claim 5, a medicament according to claim 6, a use of the invention Compounds according to claim 7, a method for complexing metal ions and a use according to claim 9.

At the 25th general meeting of the Society of German Chemists from September 10 to 16, 1995, compounds of the generic type and others were presented to the public in the form of Myxochelin C (short presentations and list of participants at the 25th General Meeting of the Society of German Chemists in Munich, 10 to 16 September 1995). In the abstract volume, Trowitzsch-Kienast et al. about Myxochelin B as a new iron transporter from the strain of the sliding bacteria Myxococcus xanthus Mx x 48 (W. Trowitzsch-Kienast, et al. 9th annual DECHEMA conference of biotechnologists, Berlin, January 1991, short reference volume, page 382) reported.

The abstract published at the GDCH conference reports on the synthetic access to myxochelin C and compounds homologous to it. Regarding the biological effect of myxochelin C, it is disclosed that myxochelin C is enormously effective as a hexadentate siderophore. The absorption of the iron-laden siderophore by enterobacteria apparently takes place after one

ORIGINAL DOCUMENTS mechanism comparable to myxochelin B (tonB dependence). The R isomer of myxochelin C shows little effectiveness.

The present invention therefore relates to compounds of the general formula I.

where R = -CH ₂ -NH-C0- (2,3-dihydroxiphenyl), CN or -CH ₂ -NH ₂ , the absolute configuration at C-2 can be both S and R and n is a natural number of 1 to 5.

The following compounds are particularly suitable according to the invention:

- if R = CN (myxochelin - nitrile)

2 myxochelin B-nitrile, n = 4; R-2 myxochelin B _R -nitrile, n = 4;

3 myxochelin D-nitrile, n = 3; R-3 myxochelin D _R -nitrile, n = 3;

- if R = -CH ₂ -NH _? (B series cheline)

R-4 myxochelin B _{R /} n = 4; 5 myxochelin DB, n = 3; R-5 Myxo¬ chelin D "-B, n = 3; Furthermore

Myxochelin C; (D, L) -1, 2, 7-triamino-tris- [N ¹ , N ² , N ⁷ - (2, 3-dihydroxy-benzoyl)] -heptane (myxochelin F).

According to the invention, the following are suitable as intermediate compounds: N, N, N-1,2,6-tris (2,3-0-dibenzyloxybenzoyl) -1,2,6-triaminohexane

(7); (D, L) -2-amino-heptanedicarboxylic acid dimethyl ester hydrochloride (la); (D, L) -2-amino- [N- (2,3-dibenzyloxybenzoyl)] -heptanedicarboxylic acid dimethyl ester (Ib); (D, L) -2-amino- [N- (2,3-dibenzyloxybenzoyl)] heptane dicarboxylic acid diamide ( _. Lc); (D, L) -2-amino-

[N- (2,3-dibenzyloxybenzoyl)] heptane nitrile (Id); (D, L) -1, 2, 6-triamino- [N ² - (2, 3-dibenzyloxybenzoyl)] heptane (le,); (D, L) - 1,2,6-triamino-tris- [N ^ N ^ N ⁶ - (2,3-dibenzyloxybenzoyl)] heptane

(lf); (L) -2, 6-diamino-bis- [N ² , N ⁶ - (2, 3-dibenzyloxybenzoyl)] - hexanecarboxamide (2a); (L) -2, 6-diamino-bis- [N ² , N ⁶ - (2,3-dibenzyloxi-benzoyl)] hexanitrile ( _. 2b); (L) -2, 6-diamino-bis- [N ² , N ⁶ -

(2,3-dihydroxybenzoyl)] hexanenitrile (myxochelin C-nitrile, 2);

(L) -2,5-diamino-bis- [N ² , N ⁵ - (2,3-dibenzyloxybenzoyl)] pentane nitrile (tetra-O-benzyl-myxochelin D-nitrile, 3a);

The present invention also relates to an iron complex of the compound of the general formula I, the iron complex having UV / VIS absorption at λ _max «= 571 nm.

The compounds according to the invention can be prepared according to the invention by a process in which starting from the amides of the amino acids lysine, ornitine or 2,3-diaminopropionic acid is reduced to the corresponding triamines using a complex hydride. The triamines formed are converted to the corresponding triamides using known coupling methods in peptide chemistry with protected 2,3-dihydroxybenzoic acids. The triamides obtained are converted by hydrogenolysis into the compounds according to the invention of the general formula (I).

Myxochelin C can be produced from L-lysinamide using known methods using DCC / HOBt coupling. The benzyl-protected 2,3-dihydroxybenzoic acid is bound amidically to the two NH ₂ groups, the primary amide is converted into the nitrile by means of crystalline triphosgene, the nitrile is reduced to the primary amine by means of sodium borohydride reduction with the aid of cobalt chloride Structure 6 leads. Figure 1 shows the reaction sequence to myxochelin B. Figure 2 shows the reaction sequence of the compound

6 to myxochelin C.

Compound 6 can be coupled into the 6-fold benzyl-protected triamide by further coupling using DCC / HOBt and an equivalent of benzyl-protected 2,3-dihydroxybenzoic acid

7 convict. 7 can be converted into the free myxochelin C by hydrogenolytic cleavage.

According to FIG. 3, compound 7 can also be obtained from L-lysinamide after reduction with lithium aluminum hydride to 1,2,6-triaminohexane and reaction of the triamine with three equivalents of the bisbenzyl-protected 2,3-dihydroxybenzoic acid.

The compounds according to the invention with a siderophoric structure are suitable for being taken up by bacteria. They overcome the bacterial cell wall. In particular, the compounds R-4 and 5 are introduced by the bacteria in an active transport process. This opens up the possibility of specifically introducing desired substances into such bacteria. These can be, for example, pharmacologically or biologically active compounds, such as pharmaceuticals, for example antibiotics, etc. However, they may also have higher molecular structures, for example nucleic acids which are able to transform the corresponding bacterium in this way, or structures of antibodies which determine Recognize, block or otherwise modify bacterial structures. The compounds to be introduced into the cell can be covalently attached, for example, to certain functional groups of the compounds according to the invention and then brought into the cell of the bacterium. The covalent bond mentioned can also be made unstable, so that this covalent bond is released again by intracellular processes and the active substance is then present in its free form in the cell. The advantage of such a coupling is that the active ingredients are brought specifically to the site of action can, so that increased active concentrations can be lowered and thus the risk of side effects can be reduced. The conjugates according to the invention can also be used for the production of corresponding medicaments.

The compounds according to the invention can be used as pharmaceuticals, it being advisable to provide an effective amount of one of the compounds of the formula I or their mixtures with pharmaceutical auxiliaries and / or carriers. The choice of aids is based, among other things, on galenical considerations, which in turn may depend on the type of application of the drugs. In principle, it is possible to apply the compounds in dissolved or solid form in appropriate dosage forms.

The medicaments can be used in particular in therapeutic approaches in which the diseases are caused by defective metal ion metabolism. This can be indicated in particular in the event of an iron or aluminum metabolism error. The pharmaceuticals according to the invention complex the metal ions, in particular iron or aluminum ions, which can then be removed from a cell. Furthermore, the compounds according to the invention are suitable as antibacterial or antiviral substances. The medicaments according to the invention can therefore be used for the treatment of bacterial and / or viral infections. The medicaments according to the invention can also be used for parasitic diseases. However, the medicament according to the invention can also be used as medicament in a form loaded with metal ions. The medicament according to the invention can remove iron and aluminum in various diseases of humans or animals, e.g. in hemosiderosis or thalassemia or also in Alzheimer's disease, in its metal ion-free form.

A suitable dosage of the medicament according to the invention can be determined by the person skilled in the art by known tests. In the form with bound iron or other metal ions, the medicament according to the invention can also be used for tumor treatment. Iron complexes are thus able to generate oxygen radicals which can attack tumors in particular.

A method using the uses according to the invention relates to the complexation of metal ions. Here, the metal ion-containing solution is brought into direct contact with solutions of the compounds according to the invention or the compounds according to the invention themselves. This method is therefore suitable for complexing, characterizing and / or removing metals from corresponding solutions containing these metal ions. Radioactive metal ions can also be complexed with the compounds according to the invention. This can serve as a starting point for the enrichment of radioactive isotopes and can be used in an analogous manner for the removal of radioactive isotopes.

The compounds according to the invention can be used for the analysis of bacteria. In particular, the presence of pathogenic enterobacteria can be quickly analyzed. For this purpose, for example, samples contaminated with pathogenic enterobacteria are incubated in an iron deficiency medium. By adding the compounds according to the invention, it is possible to selectively only stimulate a pathogenic bacterial strain to grow.

The invention is illustrated by the following examples.

example 1

N, N, N-1, 2, 6-tris- (2, 3 -0-dibenzyloxy-benzoyl) - 1, 2, 6 -triaminohexane (7): 173.9 mg (0.52 mmol) of bis-O-benzyl-protected benzoic acid, mp. 123 ° C (shown according to literature specification F. Kanai, K. Isshiki, H. Nagawana, T. Takita, T. Takeuchi, H. Umezawa, "J. Antibiot.", 37, 3987 (1985), mp. 124 ° C.) are dissolved in 8 ml of dry dimethylformamide (DMF). After the addition of 156.8 mg (1.16 mmol) of N-hydroxibenzotriazole (HOBt) and 52.9 mg (0.26 mmol) (DCC), the solution is cooled to 0 ° C. and then one hour with ice cooling and another hour stirred at RT.

245 mg of tetra-O-benzyl-protected myxochelin B, which was obtained from L-lysine according to scheme 2 above, is dissolved in 5 ml of dry DMF and added dropwise to this solution. Half-hour TLC control (dichloromethane / 5% methanol) after spraying with ninhydrin shows that after two hours there is no longer any starting material. After this time, the solvent is distilled off under an oil pump vacuum. The greenish residue is taken up in dichloromethane, first washed with 2 N hydrochloric acid, then with 1 M sodium hydrogen carbonate solution and finally with water until neutral, dried over sodium sulfate. In a water jet vacuum, the dichloromethane is removed. The residue is taken up in dry tetrahydrofuran, and the dicyclohexyl urea which precipitates is filtered off. 265.1 mg of crude product are obtained, which are fractionated by means of medium pressure column chromatography with dichloromethane / 0.37% methanol. At a flow of 6 ml / min, 7 is collected at R _t = min. After concentration, 104.2 mg of 7 are obtained (yield 37%), mp. 115 ° C.

PC behavior (Merck, Kieselgel 60 F ₂₅₄ , aluminum foil); Mobile solvent: dichloromethane with 7% methanol, R _£ = 0.42. Optical rotation: [at] ²⁵ _D = + 1.34 (c = 0.6 in CHC1 ₃ ). ^ -NMR (400 MHz, CDCl ,, see Figure 1): δ (ppm) = 8.28 (d, J = 7.4 Hz, 1 H, NH at C-2), 7.98 (t, J = 5.5 Hz, 1 H, NH at C-6), 7.81 (t, J = 3.9 Hz, 1 H, NH at Cl), 7.30 (m, 20 H, aromatic protons of the benzyl radicals ), 5.05 (m, 8 H, -CH ₂ residues of the benzyl groups), 3.99 (m, 1 H, 2-H), 3.35 (m, 1 H, 1-H _a ), 3 , 25 (m, 1 H, 1-H _b ), 3.11 (m, 2 H, 6-CH ₂ ), 1.15 to 0.98 (m, 6 H, 3,4,5-CH ₂ ). IR (CCL Figure 3): nü = 3390 (NH), 3034 (arom. H), 2939 (CH), 1662 (amide-I), 1526 cm ¹ (amide-II).

(+) -FAB-MS: Matrix: 3-nitrobenzyl alcohol m / z = 1080 (M + H) ⁺ . MS high resolution: for C ₆₉ H ₆₆ N ₃ 0 ₉ found 1080.4671 and calculated 1080.4799.

Example 2

N, N, N-1, 2, 6-tris- (2, 3-dihydroxibenzoyl) -1,2, 6-triaminohexane, myxochelin C (1):

93.8 mg (0.087 mmol) 7 are dissolved in 3 ml of methanol and 1 ml of 10% acetic acid is added. 93.8 mg of palladium with 10% activated carbon as a catalyst are added to the solution. Using a hydrogenation apparatus (an air balloon serves as the H ₂ reservoir), the mixture is first flushed with N ₂ and then hydrogenated at RT for two hours. After 90 minutes the experiment is ended, the catalyst is filtered off, the solvents are evaporated off under an oil pump vacuum (30 ° C.).

The dark-colored residue is used to separate residual catalyst and metal complexes, which had already formed from traces of metals in the solvents used, in approx. 1 ml of a solution of dichloromethane and 10% methanol, which was previously removed using Chelex 100 to remove Traces of iron were filtered, taken up and applied via a small Pasteur pipette filled with silica gel (100 mg). 1 is eluted from this mini column with a further approx. 10 ml of the solvent, the solution is then freed from the solvent under an oil pump vacuum, 1 crystallizing out in white plates.

39 mg of 1 with mp 115 ° C. are obtained, yield 83%.

PC behavior: (on silica gel plates from Merck as above): mobile phase: dichloromethane with 10% methanol and 1% glacial acetic acid, R _f = 0.29, Claims with FE- (III) -Cl ₃ solution leads to abrupt bluing of the spot, which can also be detected at 254 nm. Optical rotation: [α] ²⁰ _D = + 9.14 (c = 0.35, in methanol). * H-NMR (600 MHz, CDC1 ₃ + 5% CD ₃ OD, Figure 4): 6 (ppm) = 7.25

(m, 3 H, 3 x 6'-H of the benzoyl residues), 6.9 and 6.25 (m, 6 H, 3 x 5 '-H and 3 x 4' -H), 5.5 CH ₂ C1 ₂ residues, 4.4 (m, 1 H, 2-H), 3.65

(m, 1 H, 1-HJ, 3.59 (m, 1 H, 1-HJ, 3.43 (m, 2 H, 6-CH ₂ ), 1.9 to 1.5 (m, 6 H , 3,4,5-CH ₂ ).

Example 3

Myxochelin iron (III) complex:

If a solution of 1 in methanol is mixed with dilute iron (III) chloride solution - also in methanolic form - an immediate blue color appears: λmax »571 nm.

Example 4

To produce the enantiomeric compound myxochelin C _E , the starting point is D-lysinamide only while maintaining all the reaction conditions.

Example 5

The enantiomerically pure D- and L-ornitinamides can be used to prepare the compound which is shorter by one CH ₂ group.

Example 6

The enantiomerically pure 2, 3-diaminobutyric acid amides can be used to prepare the chelators which are shorter by two CH ₂ groups. Example 7

The enantiomerically pure 2, 3-diaminopropionic acid amides can be used to prepare the chelators which are shorter by three CH ₂ groups.

Example 8

(D, L) -2-amino-heptanedicarboxylic acid dimethyl ester hydrochloride ⁽ la ⁾ •

5.0 g (28.5 mmol) (D, L) -2-amino-heptanedicarboxylic acid (from Bachern, Switzerland) are suspended in approx. 100 ml of dry methanol. HCl _g is passed in with stirring until the solution becomes clear. The mixture is stirred overnight and then evaporated down in vacuo. The crude product is purified by flash chromatography on silica gel Si60 (Merck, Darmstadt) with n-hexane / ethyl acetate = 7: 3. 4.78 g (70%) of la are isolated.

APCI-MS: m / z (%) = 520 (50) [M + H] ^* , 542 (6) [M + Na] ^* , 317

(100). "C-NMR: (75.4 MHz, DMSO-d ₆ ): δ = 23.7 (t, C-4), 23.9 (t, C-3),

29.6 (t, C-5), 33.0 (t, C-6), 51.4 (d, C-2), 51.9 and

52.9 (q, 0-CH _3), 170.2, 173.5 (-C = 0).

For C ₉ H ₁₈ N0 ₄ C1 (MG 239.5) calc. C 45.09 H 7.52 N 5.85 Cl 14.82 found. C 45.07 H 7.66 N 5.64 Cl 15.28

Example 9

(D, L) -2-amino- [N- (2,3-dibenzyloxybenzoyl)] -heptanedicarboxylic acid dimethyl ester (lb).

1.5 g of 2,3-dibenzyloxybenzoic acid (4.5 mmol), prepared according to F. Kanai, K. Isshiki, H. Nagawana, T. Takeuchi, H. Umezawa, in "J. Antibiotics", 3_7, 3987 (1985 ) are dissolved in a flask with a drying tube in 50 ml of absolute dichloromethane and other with 1.08 g la (4.5 mmol), 1.44 g TBTU (4.5 mmol) and 1.74 g (13.5 mmol, three equiv.) Hünig base. It is stirred for 72 hours.

For the preparation, the organic phase with 1) 50 ml of 5% HCl, 2) with saturated NaHC0 ₃ solution and 3) with sat. Washed NaCl solution. It is dried with MgS0 ₄ and evaporated down in vacuo. It is cleaned by means of flash chromatography on silica gel Si60 (Merck, Darmstadt), eluent: n-hexane-ethyl acetate = 1: 1. Yield: 630 mg (27%, as a light syrup).

IR (KBr): v = 3360, (NH), 2925 (CH) 1730 (ester-CO), 1655

(Amid-I), 1560 cm ^"1 (Amid-II)

APCI-MS: m / z (%) = 520 (50) [M + H] \ 542 (6) [M + Na] ⁺ , 317 (100).

¹³ C-NMR: (75.4 MHz, CDCL ₃ ): δ = 24.5: t, c-4. : 25.1 (t, C-5)

31.7 (t, C-3), 33.7 (t, C-6), 51.6 (d, C-2), 52.3 and 52.6 (q, 0-CH ₃ ), 71 , 4 and 76.3 (t, 0-CH ₂ -phenyl), 117.4, 123.5, 124.7 (d, arom. = CH), 126.8 (s, arom. = C-), 128, 0, 128, 7, 128, 9, 129, 0 (2 x d. Arom. = C- H), 128.5 and 128.8 (s, arom. = C-), 136.5 us 136.6 (s, arom. = C-), 147.2 and 152.0 (s, arom. = C-0), 165.2 (s, amide-CO), 173.0 and 174.0 ( Ester-CO).

For C ₃₀ H ₃₃ NO ₇ (MG 519.60) calc. C 69.35 H 6.40 N 2.70 found. C 68.70 H 6.31 N 3.01

Example 10

(D, L) -2-amino- [N- (2,3-dibenzyloxybenzoyl)] heptanedicarboxylic acid diamide (lc.).

630 mg (1.2 mmol) lb_ are dissolved in 150 ml methanol, the solution is cooled to -5 ° C. NH ₃ gas is passed through the cooled solution for three hours. The still cool solution is placed in an autoclave, which is left closed at RT for four days. After careful ventilation, the solution concentrated in vacuo, yield 470 mg (80%) crystalline lc, mp. 200 to 203 ° C.

IR (KBr): v = 3390, (NH), 3180 and 3250 (NH ₂ ), 2850, 2930, 3020, 3050 (CH), 1640 and 1650 (Amid-I), 1520 cm ¹ (Amid-II) - CI-MS (NH,): m / z (%) = 490 (22) [M + H] \ 472 (18) [MH ₂ 0 + H] \

455 (55), 365 (100), 337 (56), 275 (81), 247 (48), 11 (76).

13 C-NMR: (75.4 MHz, DMSO-d ₆ ): δ (ppm) = 24.8 (t, C-4), 24.9 (t,

C-5), 32.3 (t, C-3), 34.9 (t, C-6), 52.6 (d, C-2), 70.4 and 75.1 (t, 0- CH ₂ -phenyl), 116.6, 121.6, 124.3 (d, arom. = CH), 128.2, 128.2, 128.6, 128.8 (each 2 xd, arom. = CH ), 136, 7 and 136, 8 (s, arom. = C-), 145.8 and 151.8 (s, arom. = C-0), 164.5 (s, amide-CO), 173, 6 and 174.3 (ester-CO).

C Found for C ₂₈ H ₃₁ N ₃ 0 ₅ (MW 489.58, ^'calc. C 69.69 H 6.38 N 8.58. 68.50 H 6.29 N 8.48

Example 11

(D, L) -2-amino- [N- (2,3-dibenzyloxybenzoyl)] heptane nitrile (Id).

450 mg (0.92 mmol) lc, 0.3 ml (3.68 mmol) pyridine and 200 mg (0.62 mmol) triphosgene are dissolved in 80 ml dried dichloromethane and stirred for one hour at RT. For working up, the organic phase is washed with 1) 5% HCl and 2) with sat. NaCl solution extracted. It is dried over MgS0 ₄ and evaporated down in vacuo; this gives 350 mg (84%) of pure ld.

IR (KBr): v = 3330 (-NH), 2870, 2925, 3020, 3055 (-CH), 2235

(-CN), 1650 (amide-I), 1570 (amide-II) cm ¹

-CI-MS (NHJ: m / z (%) = 454 (100) [M + H] *, 471 (8) [M + NHJ \

427 (39) [M-CN + H] ⁺ , 337 (89) [M-CN-Benzyl + H] ^* .

"C NMR: (75.4 MHz, CDCL ₃ ): δ (ppm) = 16.9 (t, C-4), 24.5

(t, C-5), 24.7 (t, C-3), 31.8 (t, C-6), 40.1 (d, C-2), 71.5 and 76.8 (t , 0-CH _? -Phenyl), 118.4 and 119.3 (s, -CN), 118.2, 123.7 and 124.8 (d, arom. = CH), 125.2, 128, 7 and 128, 78 (s, quart. arom. = C-), 129.0, 129.2, 129.3 (6C, d, arom. = CH), 136, 3 and 136, 4 (s, arom. = C-), 147.4 and 151.9 (s, arom . = C-0), 164.8

(Amide-CO).

For C ₂₈ H ₂₇ N ₃ 0 ₃ (MG 453.55) calc. C 74.15 H 6.00 N 9.26 found. C 73.66 H 6.10 N 9.20

Example 12

(D, L) -1,2,7-triamino- [N ² - (2,3-dibenzyloxybenzoyl)] heptane (le).

300 mg (0.66 mmol) id and 200 mg Co ₂ B, SW Heinzmann, B. Ganem, in J. Amer. Chem. Soc. , 1982, 6801 and 0.5 g NaBH 2 are dissolved in 10 ml THF and 40 ml methanol and stirred for two hours. For working up, 5% HC1 are added (pH 2-3), then the mixture is made basic with NH ₃ and extracted four times with 30 ml CHC1 ₃ (until the extract spotted on TLC with ninhydrin shows no positive reaction!). The chloroform solution is dried over MgSO ₄ and evaporated down in vacuo. There are 324 mg of oily diamine _. obtained as a crude product which is used in the following reaction without purification.

For C ₂₈ H ₃₅ N ₃ 0 ₃ (MG 461.61)

Example 13

(D, L) -1,2,7-triamino-tris- [N ^ N ^ N ⁷ - (2, 3-dibenzyloxi-benzoyl)] - heptane (If _. ).

300 mg of crude product le from the pre-reaction are dissolved in 50 ml of dry dichloromethane with 417 mg of TBTU (1.23 mmol), 0.43 g of dibenzyloxybenzoic acid (1.23 mmol) and 0.44 ml (2.6 mmol) of Hünig's base and stirred at rt for 72 hours. The processing takes place as described for lb. It is cleaned by means of flash chromatography with the mobile phase n-hexane / ethyl acetate = 1: 1. 311 mg If are obtained, 43% based on Id from stage 4.

IR (Kbr): v = 3370 (-NH), 2850, 2915, 3020, 3055 (CH), 1630

(Amide-I), 1565 cm ^"1 (Amide-II).

ESI-MS: m / z (%) = 1094.4 (100) [M + H] \ 1116.5 (65) [M + Na] \ 1132.4 (36) [m + K] ⁺ .

"C-NMR: (75.4 MHz, CDCL ₃ ): δ (ppm) = 25.7 (t, C-5), 26.9 (t, C-4), 29.2 (t, C- 6), 32.3 (t, C-3), 39.7 (t, C-7), 43.5 (t, Cl), 49.9 (d, C-2), 71.4, 71 , 4, 71.4, 75.9, 76.2, 76.5 (t, 0-CH ₂ -Phe), 117.0, 117.1, 117.2 (d, arom. = CH-), 123.4, 123.6, 123.7 and 124.4, 124.5, 124.6 (d, arom. = CH-), 127.2, 127.7, 128.0 (s, quart. Arom . = C-), 127.8, 127.9, 127.9, 128.5, 128.5, 128.7, 128.7, 128.8, 128.80, 128.85, 128.85, 128.90, 128.90, 128.93, 128.93 (all as d, arom. = CH-), 136.57, 136.6, 136.6, 136.63, 136.7, 136.7 (s, quart. arom. = C-), 146.9, 147.0, 147.1 (s, quart. = C-0), 151.9, 152.0, 152.0 (s, quart. = C-0), 165.2, 165.4, 165.9 (s, amide-CO).

For C ₇₀ H ₆₇ N ₃ O ₉ (MW 1094.33) calc. C 76.83 H 6.17 N 3.84 found. C 75.91 H 6.23 N 4.19

Example 14

(D, L) -1,2,7-triamino-tris- [N ¹ , ^, ^ - (2, 3-dihydroxi-benzoyl)] - heptane (Myxochelin F, 1).

113 mg (0.1 mmol) if _. are dissolved in 20 ml of THF and 30 ml of methanol and hydrated with Pd / CH ₂ for two hours at normal pressure. It is filtered through diatomaceous earth, the solution is evaporated down in vacuo, yield 50 mg, 87%.

IR (KBr): v = 3360 (NH), 2920, 2850 (CH), 1640 (amide-I), 1580

(Aromatic), 1540 cm ^"1 (Amide-II).

ESI-MS: m / z (%) = 576 (100) [M + Na] ⁴ . "C NMR: (100.6 MHz, CD ₃ OD): δ (ppm) = 26.9 (t, C-4), 27.8 (t, C-5), 30.3 (t, C -6), 33.1 (t, C-3), 40.4 (t, C-7), 44.6 (t, Cl), 51.2 (d, C-2), 116.9, 116.9, 117.0 (s, quart. Arom. = C-), 118.7, 118.9, 119.0, 119.6, 119.7, 119.7 (all d, arom. = CH -), 147.3, 147.3, 147.4 (s, aroma = C-0-), 150.2, 150.2, 150.3 (s, aroma = C-0-), 171 , 5, 171.8, 172.0 (s, amide-CO).

For C ₂₈ H ₃₁ N ₃ O _g (MW 553.58) calc. C 60.75 H 5.64 N 7.59

The following examples relate to the siderophore Myxochelin- C- nitrile _(2.) And a method for its preparation.

The substance is described below on the basis of its production and its spectroscopic properties:

Example 15

It is obtained from: (L) -2, 6-diamino-bis- [N ² , N ⁶ - (2, 3-dibenzyloxi-benzoyl)] -hexanecarboxamide ( _. 2a), which itself can be purchased L-lysinamide is shown. The spectroscopic features of .2a are as follows. :

[α] ²⁰ _D = - 9 (c = 0.5 in acetone).

IR (KBr): v = 3371, 3200 (NH), 3032, 2930 (CH), 1651 (Amid-I),

1526 cm ^"1 (Amide II).

¹³ C-NMR: (75.4 MHz, CDCl ₃ ): δ (ppm) = 23.03 (t, C-4), 28.9 (t, C-5), 30.6 (t, C- 3), 39.1 (t, C-6), 53.1 (d, C-2), 71.3, 71.4, 76.3, 76.4 (t, 0-CH _? -Phenyl) , 116.9, 117.4, 123.0, 123.3, 124.4, 124.4 (d, arom. = CH-),

126.7, 127.3 (s, quarter aroma = C-) 127.6, 127.6,

127.8, 128.2 (d, aroma. = CH-), 128.3 (d, 10 x aroma. = CH-), 128.7 (d, 5 x aroma. = CH-) 128.9 (d, arom. = CH-), 136.2, 136.3, 136.4, 136.4 (s, quart. arom. = C-), 146.8, 146.9, 151.7, 151.7 (s, = C-0-), 165.1, 165.6 (s, sec.amide-C =), 173.7 (s, prim Amide CO)

For C ₄₈ H ₄₇ N ₃ 0 ₇ (MW 777.34) calc. C 74.16 H 6.10 N 5.41 found. C 72.60 H 6.07 N 4.60

Example 16

(L) -2,6-Diamino-bis- [N ² , N ⁶ - (2,3-dibenzyloxybenzoyl)] hexanitrile (2b).

0.98 g (1.2 mmol) 2a. are dissolved in 15 ml of absolute dichloromethane and mixed with 0.29 ml of pyridine and 176 mg of triphosgene. After working up - as described for ld - 670 mg 2b are obtained (73.6%).

[α] ²⁰ _D = - 16.1 (c = 1 in CHC1 ₃ ).

IR (KBr): v = 3367 (NH), 2866, 2927, 3031, 3064 (CH), 2230 (CN),

1661 (amide-I), 1520 cm ^"1 (amide-II).

"C-NMR: (75.4 MHz, CDC1 ₃ ): δ (ppm) = 22.7 (t, C-4), 28.4 (t,

C-5), 32.1 (t, C-3), 39.0 (t, C-6), 40.3 (d, C-2), 71.3, 71.4, 76.4, 76.6 (t, 0-CH ₂ -Phe), 117.0, 117.9 (d, arom. = CH-), 118.5 (s, -CN), 123.3, 123.4, 124 , 4, 124.5 (d, arom. = CH-), 127.6, 127.6, 128.4, 128.4 (d, arom. = CH), 128.3, 128.4 (s, fourth aroma = C-) 128.7 (4 x C), 128.8 (8 x C), 129.0 (4 x C, all d, 16 aroma = CH- from Bn groups), 135 , 9, 136.0, 136.2, 136.4 (s, arom. = C- from Bn groups), 146.8, 147.1 (s, quart. Arom. -0-C =), 151 , 6, 151.7 (s, quart. Arom. 0-C =), 164.5, 165.0 (s, amide-0 =).

For C ₇₅ H ₄₅ N ₃ 0 ₆ (MG 759.91) calc. C 75.87 H 5.97 N 5.53 found. C 72.18 H 5.63 N 4.16 Example 17

(L) -2, 6-diamino-bis- [N ² , N ⁶ - (2, 3-dihydroxi-benzoyl)] hexanitrile (myxochelin C-nitrile, 2_).

There are 90 mg (0.12 mmol) _. 2b hydrides under standard conditions. After filtration through diatomaceous earth and concentration in vacuo, 45 mg (95%) 2 are isolated.

[a] ²⁰ _D = - 12 (c = 1 in methanol). MG 399.43

The following examples relate to myxochelin D-nitrile (3_) and a process for its production.

Example 18

3_ becomes - as for 2 _. shown - obtained from the tetra-benzyl-protected myxochelin D-nitrile (3_a) by hydrogenolytic cleavage on Pd / C using H ₂ . 3_a is characterized as follows:

Example 19

(L) -2, 5-Diamino-bis- [N ² , N ⁵ - (2, 3-dibenzyloxybenzoyl)] pentane nitrile (tetra-O-benzyl-Myxochelin D-nitrile, 3a).

Mp 134-136 ° C [α] ²⁰ _D = 18.4 (c = 1 in methanol).

IR (KBr): v = 3360 (NH), 3015, 3065, 2900, 2925, 2860 (CH), 1650

(Amide-I), 1520 cm ^"1 (Amide-II).

Cl- (+) -MS: m / z (%) = 746 (100) [M + H] ⁴ .

"C-NMR: (75.9 MHz, CDCl ₃ ): δ (ppm) = 25.4 (t, C-4), 30.0 (t, C-3), 38.5 (t, C- 5), 40.3 (d, C-2), 71.3, 71.4, 76.5, 76.7 (t, 0-CH ₂ -Phe), 117.1, 117.9 (d, arom. = CH-), 118.3 (s, -CN), 123.4, 123.5, 124.4, 124.5 (d, arom. = CH-), 127.6, 127.7 (d, arom. = CH-), 128.3, 128.4 (s, quart. Arom. = C-) 128.7, 128.8, 128, 9 129.1 (18 C, d, arom. = CH-), 135.8, 136.2, 136.3, 136.4 (s, quart. Arom. = C ~), 146.9, 147, 1 (s, quart. Aroma. -0-C =), 151.6, 151.63 (s, quart. Aroma. -0-C =), 164.4, 165.0 (s, sec. Amide- CO).

For C ₄₇ H ₄₃ N ₃ 0 ₆ (MW 745.87) calc. C 75.68 H 5.81 N 5.63 found. C 75.66 H 5.80 N 5.36

Example 20

(L) -2, 5-diamino-bis [N ^2, N ⁵ - (2,3-dihydroxy benzoyl)] pentane-nitrile _(. Myxochelin- D-nitrile, 3).

50 mg (7.7 x 10 ^"5 mol) of L-2,5-diamino-bis- [N ² , N ⁵ - (2,3-dibenzyloxybenzoyl)] -pentanitrile (3_a) are hydrogenated under standard conditions Filtration through diatomaceous earth and concentration in vacuo give 24 mg (93.4%) 3_.

[o;] ²⁰ _D = - 14.5 (c = 0.9 in methanol).

IR (KBr): v = 3360 (NH), 2925 (CH), 2230 (CN) 1630 (amide-I),

1525 cm ^'1 (amide-II).

¹³ C-NMR: (125.6 MHz, CD ₃ OD): δ (ppm) = 26.8 (t, C-4), 31.0 (t,

C-3), 39.4 (t, C-5), 41.5 (d, C-2), 116.1, 116.7 (s, quart. Arom. -C =) 118.7, 119 , 1 (d, arom. = CH-), 118.7 (s, -CN), 119.6, 119.7, 120.0, 120.4 (d, arom. = CH-), 147.3 , 147.4, 150.2, 150.3 (s, quart. -0-C =), 171.1, 171.7 (s, sec. Amide-CO).

'H-NMR: (500 MHz, CD ₃ OD): δ (ppm) = 1.02 (quin., J _x = 14.8 Hz,

J ₂ = 7.4 Hz 2H, 4-CH ₂ -), 125 (9 lines, J _x = 15.5, J ₂ = 7.8, J ₃ = 7.7 Hz, 2H, 3-CH ₂ - ), 2.67 (dd, J _λ = 6.6, J ₂ = 7.0 Hz, 1H, 2-H), 5, 91 (dd, J, = 7, 8, J _z = 8.1 Hz , 1H, meta-H _aroma ), 5.95 (dd, J _x = 8.1, J _? = 8.2 Hz, 1H, meta-H _aron ,), 6.13 (dd, J ₁ = 1, 2, J ₂ = 7.8 Hz, 1H, para¬ t _em ), 6.17 (dd, J, = 1.2, J ₂ = 7.8 Hz, 1H, para H _aroma ), 6.41 (dd, J ₁ = 1.5, J ₂ = 8.1 Hz, 1H, ortho-H _aroma ), 6.45 (d, J _α = 1.1, J ₂ = 8 , 1 Hz, 1H, ortho-

^H arom.) ^•

ESI- (+) -MS: m / z (%) = 408 (100) [M + Na] ⁴ .

For C ₁₉ H ₁₉ N ₃ 0 ₆ (MG 385.39) calc. C 59.36 H 4.72 N 10.93 found. C 57.10 H 5.44 N 9.78

The following example relates to myxochelin D _R -nitrile R-3, and a process for its preparation which follows the same route as described for 3_.

Example 21

The precursor R-3a shows the same spectroscopic properties as 3_a, but has a rotation value of:

[a] ²⁰ _D = + 16.9 (c = 1 in methanol).

(R) -2,5-diamino-bis- [N ² , N ⁵ - (2,3-dihydroxybenzoyl)] pentane nitrile (myxochelin D _R nitrile, R-3).

48 mg of R-3a are hydrogenated under standard conditions at normal pressure. After filtering off over kieselguhr and concentrating i.V. 22 mg (88.7%) of R-3 are obtained. See the description for R-3 for the spectroscopic properties.

[α] ^2D _D = + 16 (c = 1 in methanol).

The following example relates to the enantiomer of the natural product myxochelin B, the myxochelin B _R (R-4) and a process for its preparation.

Example 22

The starting point is R-2b, the spectroscopic properties of which, with the exception of the rotational value, are the same as for _. 2 B are (W. Trowitzsch-Kienast, H. Irschik, V. Wray, H. Reichen¬bach, G. Höfle, Liebigs Ann. Chem. 1996, in preparation). The rotation value for R-2b is:

[ot] _D = + 18.5 (c = 1 in CHC1 ₃ ).

The nitrile R-2b is converted into the primary amine by means of NaCNBH ₃ , which is hydrogenated by standard hydrogenation at RT and under normal pressure for two hours on a Pd / C catalyst:

(R) -1,2,6-triamino-bis- [N ² , N ⁶ - (2,3-dihydroxybenzoyl)] hexane hydrochloride (Myxochelin B _R hydrochloride, R-4).

[ot] ^2D _D = + 7 (c = 0.5 in 6N HC1). (The natural product myxochelin B has: [a] ² ° _D = - 8 (c = 1 in 6N HC1) (W. Trowitzsch-Kienast, H. Irschik, V. Wray, H. Reichenbach, G. Höfle, Liebigs Ann. Chem. 1966, in preparation).

¹³ C-NMR: (75.4 MHz, DMSO-d ₆ ): δ (ppm) = 22.9 (t, C-4), 28.7 (t, C-5), 31.3 (t, C-3), 38.6 (t, C-6), 48.7 (d, C-2), 44.0 (t, Cl), 115.7, 115.7 (s, quart. Aroma. C), 114.1, 115.8, 116.0, 117.2, 117.8, 118.4 (d, arom. = CH-), 147.0, 147.6, 151.6, 153, 6 (s, quart. -0-C =), 169.3, 169.9 (s, sec. Amide-CO).

FAB- (+) -MS: m / z (%) = 404 (100) [M + H] ⁴ .

C ₂₀ H ₂₅ N ₃ O ₆ (MG 403, 44)

effect

The effect of 1 is proven by means of a bioassay.

The following strains of enterobacteria (gram-negative bacteria), which have a defect in the iron transport system and therefore cannot absorb the iron, are so well supplied with iron by 1 in the concentration of 5 μg / disc that the Strains in a medium depleted of iron nevertheless show enormous growth zones (in mm):

Salmonella typhimurium (32), E. coli (30), Klebsiella pneumonia (33), Pseudomonas aeruginosa strain 6609 (34), strain 648 (30), strain 201 (32), strain K 437 (34).

The test conditions are published in R. Reissbrodt, L. Heischisch, U. Möllmann, W. Rabsch, H. Ulbricht, "Biometais", 6, pages 155 to 162 (1993) and R. Reissbrodt "and W. Rabsch, "Zbl. Bakt. Hyg. ", A 268, pages 306 to 317, (1988).

Myxochelin C and Myxochelin C _R have antiviral activities against cytomegalon viruses from strain AD-169. The IC ₅₀ values for the active substances are 0.7 µg / ml for myxochelin C and 1 µg / ml for myxochelin C _B.

Claims

Expectations

Compound of the general formula (I)

where R = -CH ₂ -NH-CO- (2, 3-dihydroxiphenyl), CN or -CH ₂ - NH ₂ , n = 1 to 5.

2. A compound according to claim 1, wherein R = CN (myxochelin nitriles) and the compounds

2 myxochelin B-nitrile, n = 4; R-2 myxochelin B _R -nitrile, n = 4; 3 myxochelin D-nitrile, n = 3; R-3 comprises myxochelin D _R -nitrile, n = 3,

where R = -CH ₂ -NH ₂ (cheline of the B series) and the compounds

R-4 myxochelin B _R , n = 4; 5 myxochelin DB, n = 3; R-5 comprises myxochelin D _R -B, n = 3; Furthermore

Myxochelin C; (D, L) -1, 2,7-triamino-tris- [N ¹ , N ² , N'- (2,3-dihydroxy-benzoyl)] heptane (Myxochelin F). Links

N, N _# N-1,2,6-tris (2,3-O-dibenzyloxybenzoyl) -1,2,6-triaminohexane; (D, L) -2-amino-heptanedicarboxylic acid dimethyl ester hydrochloride; (D, L) -2-amino- [N- (2,3-dibenzyloxybenzoyl)] heptanedicarboxylic acid dimethyl ester; (D, L) -2-amino- [N- (2,3-dibenzyloxy-benzoyl)] -heptanedicarboxylic acid diamide; (D, L) -2- amino- [N- (2, 3 -dibenzyloxybenzoyl)] heptane nitrile; (D, L) - 1,2,7-triamino- [N ² - (2,3-dibenzyloxybenzoyl)] heptane;

(D, L) -1,2,7-triamino-tris- [N ^ N ^ N ⁷ - (2,3-dihydroxybenzoyl)] -heptane; (L) -2, 6-diamino-bis- [N ² , N ⁶ - (2, 3 -dibenzyloxybenzoyl)] hexane carboxamide; (L) -2, 6-diamino-bis- [N ² , N ^G -

(2,3-dibenzyloxybenzoyl)] hexanenitrile; (L) -2, 6-diamino-bis- [N ² , N ⁶ - (2, 3 -dihydroxy-benzoyl)] hexanitrile (myxochelin C-nitrile) or (L) -2, 5-diamino-bis- [N ² , N ^B - (2, 3 -dibenzyloxybenzoyl)] pentane-nitrile (tetra-O-benzyl-Myxochelin D-nitrile) as intermediates for the preparation of compounds according to claim 1 and / or 2.

Iron complex of the compound according to one of claims 1 to 3, characterized in that the iron complex has a UV / VIS absorption at λ _max «= 571 nm.

Process for the preparation of the compounds according to one of the

Claims 1 to 3, wherein the amides of the amino acids lysine, ornitine, asparaginamide

(Myxochelin E) or 2, 3-diaminopropionic acid amide with a complex hydride can be reduced to the corresponding ones

Triamines,

the resulting triamines are converted to the corresponding triamides using coupling methods of peptide chemistry with protected 2, 3-dihydroxybenzoic acids and

The triamides obtained are converted into the compounds according to one of claims 1 or 2 by hydrogenolysis. 6. Conjugate of a compound according to at least one of claims 1 to 3 and a pharmaceutically and / or biologically active substance such as a drug.

7. Medicaments containing, in addition to conventional pharmaceutical auxiliaries and / or carriers, an effective amount of at least one of the compounds according to claims 1 to 3 and / or 5

8. Use of a compound according to at least one of claims 1 to 3 for the manufacture of a medicament for the treatment of diseases which are correlated with defective metal ion metabolism, in particular iron or aluminum metabolism, or for the discharge of metal ions, in particular iron or aluminum ions, from cells and / or for the treatment of bacterial, viral and / or parasitic infections and for the treatment of tumors.

9. A method for complexing metal ions, in particular iron ions, by adding a metal ion-containing solution with a compound according to at least one of claims 1 to 3.

10. Use of a compound according to at least one of claims 1 to 3 for complexing, characterizing and / or removing metals from the solutions containing the corresponding metal ions.

11. Use according to claim 10, wherein the metal ions are radioactive metal ions.

12. A method for the analysis of bacteria, in particular pathogenic enterobacteria, wherein the samples contaminated with bacteria are incubated in an iron deficiency medium and, by adding a compound according to at least one of claims 1 to 3, selectively the bacteria which are able to grow them To make connections.