DE19643764A1 - Glycoconjugates of Modified Camptothecin Derivatives (20-0 Link) - Google Patents

Abstract

Disclosed are glycoconjugates from modified camptothecin derivates, where at least a glucidic component is linked to the 20-hydroxyl group of a camptothecin derivate. Also disclosed are their preparation and their use as pharmaceutical products, especially against cancer diseases.

Description

The present invention relates to glycoconjugates of camptothecin derivatives, in which have at least one carbohydrate component via suitable spacers and Spacers linked to the 20-hydroxyl group of a camptothecin derivative is. The invention further relates to methods for producing the fiction, ge moderate compounds and their use as medicines, especially in Cancer related.

20 (S) -camptothecin is a pentacyclic alkaloid developed by Wall et al. was isolated (J. Am. Chem. Soc. 88, 3888 (1966)). It has a high anti-tumor effect potential in numerous in vitro and in vivo tests. Unfortunately, however, failed the realization of the promising potential in the clinic at toxicity and Solubility problems.

One opened by opening the E-ring lactone and formation of the sodium salt water-soluble compound obtained in a pH-dependent equilibrium with the ring-closed form. Clinical studies have also been carried out here so far not to success.

About 20 years later, it was found that the biological activity decreased to one Enzyme inhibition of topoisomerase I is due. Since then, the Research activities again intensified to be more tolerable and effective in vivo Find camptothecin derivatives.

Salts from A-ring and B-ring were used to improve water solubility modified camptothecin derivatives and 20-O-acyl derivatives with ioni described groups (Vishnuvajjala et al. US 4943579). The latter The prodrug concept was later also modified on camptothecin derivatives transferred (Wani et al. WO 9602546). The 20-O-acyl prodrugs described  However, they have a very short half-life in vivo and become very fast split to the main body.

We now found it surprising that the attachment of carbohydrate derivatives for example via peptide spacers to the 20-hydroxyl group of A- and / or B- Ring-modified camptothecin derivatives to a compound class with high interesting properties:

• - By the ester-like connection of the carrier residues to the 20-hydroxyl group is the lactone ring of Camptothecin-De derivatives stabilized.
• - The conjugates obtained in this way show a high activity towards in vitro Tumor cell lines and tumor xenografts.
• - They clearly show a comparison to the underlying toxophores higher tolerance and tumor selectivity as well as an improved dissolution sensitivity, especially in aqueous media.
• - In vivo they show excellent therapeutic efficacy over several Dose levels.
• - They are much more stable than that in extracellular medium and in blood previously described 20-O-acyl prodrugs of camptothecin.

The invention relates to compounds of the general formula (I)

A-Cp-B (I)

wherein
Cp for a group of formulas

wherein
R¹, R², R³ and R⁴ independently of one another can denote hydrogen alkyl with up to 3 carbon atoms, halogen, amino, hydroxyl or nitro or
R² and R³ together form a group of the formula

mean where
m can have the values 1 or 2, and
R⁵ for H, -CH₂CH₃, -CH₂-O- *, -CH₂-NH *,

stands,
or for -CH₂-N (CH₂CH₃) R⁶ or

stands,
wherein R⁶ means arylmethyl or hetarylmethyl,

wherein
R⁷ and R⁸ are defined as R² and R³ and can be the same or different with them,

wherein
R⁹ represents hydrogen or -CH₂-N (CH₃) ₂ and
R¹⁰ represents hydrogen or ethyl,
or

wherein
R¹¹ and R¹² are defined as R² and R³ and can be the same or different with them,
or

wherein
R¹³ and R¹⁴ are defined as R² and R³ and can be the same or different with them,
where Cp is linked to A at the positions marked with # and B to the positions marked with *,
A is a residue of the formula

means, where 1 (n + o + p) 3 applies,
B is hydrogen or a radical of the formula

means, where 0 (q + r + s) 3, where
M¹ and M² independently represent a bridging group whose main chain comprises up to 21 atoms in a linear sequence,
L¹, L², L³, L⁴, L⁵ and L⁶ independently of one another stand for liner groups, as are commonly used in glycoconjugate chemistry (see review article Lee YC and Lee R. in Lectins and Cancer 1991, 53-69, ed. By Gabius HJ and Gabius S., Springer-Verlag),
Sp¹, Sp², Sp³, Sp⁴, Sp⁵ and Sp⁶ independently represent arylene with up to 10 carbon atoms or for alkylene with up to 8 carbon atoms, which are each optionally substituted, and
K¹, K², K³, K⁴, K⁵ and K⁶ independently of one another for a radical of the formula (II)

stand in what
E is methyl, hydroxymethyl, alkoxymethyl with up to 6 carbon atoms, acyloxymethyl with up to 6 carbon atoms or a radical of the formula -CH₂-D, wherein
D represents a radical of the formula (II),
R¹⁵, R¹⁶ and R¹⁷ independently of one another hydrogen, hydroxy, optionally substituted by hydroxyl-substituted alkoxy having up to 6 carbon atoms, optionally substituted by alkyl or acyl having up to 6 carbon atoms, amino, halogen, sulfate or a group of the formulas

mean what
R¹⁸ and R¹⁹ independently of one another represent hydroxyl or alkoxy having up to 6 carbon atoms or amino, which is optionally substituted by alkyl having up to 6 carbon atoms, and
u and v can independently assume the values 0, 1, 2, 3 or 4, in particular the values 1, 2, 3 or 4
or
R¹⁵, R¹⁶ and R¹⁷ independently represent a radical of the formula (II)
or
two of the radicals R¹⁵, R¹⁶, R¹⁷ together represent an epoxy group,
and their isomers, mixtures of isomers and salts.

The term “alkyl groups” is intended, unless otherwise stated, in the sense of the invention dung straight-chain, branched, cyclic and cycloalkyl radicals containing Include alkyl radicals. This definition should also apply to everyone else Residues containing alkyl groups apply, such as, for example, alkoxy, acyl, etc.

The terms given in the definition of R⁶ arylmethyl and Hetarylmethyl can mean, for example, phenylmethyl or pyridylmethyl.

Preferred compounds of the general formula (I) are those in which K¹, K², K³, K⁴, K⁵ and K⁶ can independently of one another represent a radical of the formula (II), where
E denotes methyl, hydroxymethyl, methoxymethyl or acetoxymethyl,
R¹⁵ is hydrogen, hydroxyl, methoxy or a group of the formulas

means what
u and v can independently assume the values 1 or 2 and
R¹⁸ and R¹⁹ independently of one another represent hydroxyl or alkoxy having up to 4 carbon atoms,
or
R¹⁵ represents a radical of the formula (II),
R¹⁶ is hydrogen, hydroxyl, halogen, alkoxy with up to 4 carbon atoms, sulfate or a group of the formulas

means what
u and v can independently assume the values 1 or 2 and
R¹⁸ and R¹⁹ independently of one another represent hydroxy or alkoxy having up to 4 carbon atoms or amino, which is optionally substituted by alkyl having up to 4 carbon atoms,
R¹⁷ hydroxyl, alkoxy with up to 4 carbon atoms, which is optionally substituted by hydroxy, optionally substituted by alkyl or acyl with up to 4 carbon atoms, or a group of the formulas

means what
u and v can independently assume the values 1 or 2 and
R¹⁸ and R¹⁹ independently of one another represent hydroxyl or alkoxy having up to 4 carbon atoms,
or in what
R¹⁵ and R¹⁶ together represent an epoxy group,
and their isomers, mixtures of isomers and salts.

According to a particularly preferred embodiment, the carbohydrate structure comprises stones K¹, K², K³, K⁴, K⁵ and / or K⁶ each have a maximum of two monosaccharide structures stones.

Also preferred are compounds of the general formula (I) in which Sp 1, Sp², Sp³, Sp⁴, Sp⁵ and Sp⁶ independently of one another for aryls with up to 10 Koh lenstoffatomen can stand, each with a group K¹, K², K³, K⁴, K⁵ or K⁶ and L¹, L², L³, L⁴, L⁵ or L⁶ is linked and the one or more multiple times by hydroxy, carboxy, carboxyalkyl with up to 4 carbon atoms, Nitro, cyano, halogen, alkyl with up to 4 carbon atoms, haloalkyl with up to 4 carbon atoms or by alkoxy with up to 4 carbon atoms is substituted, as well as their isomers, isomer mixtures and salts.

Particularly preferred substituents of Sp¹, Sp² and / or Sp³ are halogen, Nitro, alkyl with up to 4 carbon atoms, alkoxy with up to 2 carbons atoms, -OCF₃ and / or -CF₃.

Compounds of the general formula (I) are also preferred.
wherein
L¹, L², L³, L⁴, L⁵ and L⁶ independently

or

mean where
R²⁰ represents chlorine or hydroxyalkylamino with up to 6 carbon atoms.

Also preferred are compounds of the general formula (I), in which M 1 and M² can independently represent a peptide which has a Amino function with L¹, L², L³, L⁴, L⁵ and / or L⁶ is linked via a Acyl function is linked to Cp and its amino acid building blocks if necessary Protective groups can wear. Mono-, di- and Tripeptides, especially mono- and dipeptides.

The amino acid units are preferably selected from the group glycyl, Alanyl, valyl, leucyl, lysyl, seryl, glutamyl, threonyl, asparagyl, isoleucyl, Diaminopropionyl, diaminobutyryl, histidyl and / or omithyl.

The amino acid building blocks glycyl, alanyl, valyl, leucyl, Lysyl, Seryl, Asparagyl, Histidyl and / or Glutamyl.

The compounds of the invention can in stereoisomeric forms, for example as enantiomers or diastereomers, or as mixtures thereof, for example as a racemate. The invention relates to both the pure stereoiso mere as well as their mixtures.

Mixtures of stereoisomers can, if necessary, be introduced in a known manner into the stereoisomerically uniform constituents are separated, for example by Chromatography or by crystallization processes.

The stereochemistry at the anomeric center of the carbohydrate building blocks K¹, K², K³, K⁴, K⁵ and / or K⁶ can be α or β. Furthermore, you can in the D or  L shape. Due to the stereochemistry at the other centers, the gluco, manno, galacto, gulo, rhamno or fuco configuration.

The amino acid building blocks can each be in the D or L form.

The camptothecin building block Cp can be in the 20 (R) or in the 20 (S) configuration or as a mixture of these two steroisomeric forms. The is preferred 20 (S) configuration.

Preferred examples of the camptothecin building block are:

The compounds according to the invention can also be present in the form of their salts. In general, here are salts with organic or inorganic bases or Acids and internal salts called.

The acids that can be added preferably include halogen water Substance acids, such as B. the hydrochloric acid and hydrobromic acid, ins especially hydrochloric acid, also phosphoric acid, nitric acid, Schwe rock acid, mono- and bifunctional carboxylic acids and hydroxycarboxylic acids, such as e.g. B. acetic acid, trifluoroacetic acid, maleic acid, malonic acid, oxalic acid, glucon acid, succinic acid, fumaric acid, tartaric acid, citric acid, salicylic acid, sorbine acid and lactic acid and sulfonic acids, such as. B. p-toluenesulfonic acid, 1,5-naphthalenedisulfonic acid or camphorsulfonic acid.

Physiologically acceptable salts can also be metal or ammonium salts of the compounds of the invention, which be a free carboxyl group to sit. Z are particularly preferred. B. sodium, potassium, or magnesium Calcium salts, as well as ammonium salts, which are derived from ammonia or organic amines such as ethylamine, di- or triethylamine, di- or Triethanolamine, dicyclohexylamine, dimethylaminoethanol, arginine, lysine, Ethylenediamine or phenethylamine.

The glycoconjugates according to the invention can be produced, for example by linking modified camptothecin derivatives with activated ones Carboxyl components, which in turn are parts of protected amino acids, peptides or carbohydrate-modified peptides.

The invention therefore further relates to a method for producing verbin of the general formula (I), characterized in that Verbin of the general formula (III)

H _B -Cp-H _A (III)

wherein Cp has the meaning given above and the hydrogen atom H _{A is located} at the position marked with # and H _B at the *, optionally after replacing H _B by a protective group, with an activated carboxyl component M¹a corresponding to the radical M¹ defined above, which optionally carries protective groups, in a suitable solvent, if appropriate in the presence of a base, reacted by customary methods, optionally selectively cleaves off one, more or all of the protective groups of M 1 using known methods and the product is reacted with compounds of the general formula (IV)

K¹-Sp¹-L¹a (IV)

wherein K¹ and Sp¹ are as defined above and L¹a is a reactive precursor of the group L¹, reacted, optionally splitting off the protective groups and gradually introducing different groups K²-Sp²-L²- and K³-Sp³-L³- in a comparable manner can, and that, if a carbohydrate component is to be linked to the position marked with *, the protective group which replaces H _B is optionally split off by known methods, and in the manner described above the remainder of M² and, as desired, the remainder of the Introduces formulas K⁴-Sp⁴- L⁴-, K⁵-Sp⁵-L⁵- and K⁶-Sp⁶-L⁶-
or that, if M 1 and / or M 2 is a peptide, a first amino acid residue in the form of the corresponding carboxyl component, optionally bearing protective groups, is introduced in a comparable manner by conventional methods, optionally protecting groups are removed, and optionally amino acid residues bearing protective groups, as indicated above, are residues of the formulas K¹-Sp¹-L¹-, K²-Sp²-L²-, K³-Sp³-L³-, K⁴-Sp⁴-L⁴-, K⁵-Sp⁵-L⁵- and / or K⁶-Sp⁶-L⁶- and if necessary cleaves protective groups.

The reactions can take place at different pressure and temperature conditions, for example 0.5 to 2 bar, or -30 to + 100 ° C, in suitable solvents such as dimethylformamide (DMF), tetrahydrofuran (THF), dichloromethane, chloro form, lower alcohols, acetonitrile, dioxane, water or in mixtures of mentioned solvents are carried out. Usually reactions are in DMF or THF / dichloromethane at room temperature and normal pressure is preferred.

For the activation of the carboxyl groups come those in peptide chemistry knew coupling reagents such as z. B. in Jakubke / Jeschkeit: amino acids, Peptides, proteins; Verlag Chemie 1982 or Tetrahedr. Lett. 34, 6705 (1993) are described in question. For example, acid chlorides are preferred, N-carboxylic acid anhydrides or mixed anhydrides.  

The formation of is also suitable for activating the carboxyl groups Adducts with carbodiimides e.g. B. N, N'-diethyl, N, N'-diisopropyl, N, N'-dicyclo hexylcarbodiimide, N- (3-dimethylaminopropyl) -N'-ethyl-carbodiimide hydrochloride, N-Cyclohexyl-N '- (2-morpholinoethyl) carbodiimide-metho-p-toluenesulfonate, or Carbonyl compounds such as carbonyldiimidazole, or 1,2-oxazolium compounds such as 2-ethyl-5-phenyl-1,2-oxazolium-3-sulfate or 2-tert-butyl-5-methyl-isoxa zolium perchlorate, or acylamino compounds such as 2-ethoxy-1-ethoxycarbonyl- 1,2-dihydroquinoline, or propanephosphonic anhydride, or isobutylchloroform, or benzotriazolyloxy-tris (dimethylamino) phosphonium hexafluorophosphate, 1-hydroxybenzotriazole or hydroxysuccinimide ester. Furthermore, the Amino acid component also used in the form of a Leuchs anhydride will.

Triethylamine, ethyldiisopropylamine, pyridine, N, N-dimethylaminopyridine or others can be used.

As protective groups for possible further reactive functions in the camptothecin part or for third-party functions of the amino acids can be used in peptide chemistry known protecting groups, for example from urethane, alkyl, acyl, ester or Amide type can be used.

Amino protecting groups in the context of the invention are the usual ones in peptide chemistry I used amino protecting groups.

These preferably include: benzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, 2,4-dimethoxybenzyloxycarbonyl, 4-methoxy benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, 2-nitro- 4,5-dimethoxybenzyloxycarbonyl, methoxycarbonyl, ethoxycarbonyl, tert-butoxy carbonyl (Boc), allyloxycarbonyl, vinyloxycarbonyl, 3,4,5-trimethoxybenzyloxy carbonyl, phthaloyl, 2,2,2-trichloroethoxycarbonyl, 2,2,2-trichloro-tert-butoxy carbonyl, menthyloxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxy carbonyl (Fmoc), formyl, acetyl, propionyl, pivaloyl, 2-chloroacetyl, 2-bromo acetyl, 2,2,2-trifluoroacetyl, 2,2,2-trichloroacetyl, benzoyl, benzyl, 4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl, phthalimido, isovaleroyl or benzyloxy methylene, 4-nitrobenzyl, 2,4-dinitrobenzyl, 4-nitrophenyl or 2-nitrophenylsul fenyl. The Fmoc group and the Boc group are particularly preferred.  

Preferred carboxyl protecting groups are linear or branched C₁-C₄ alkyl esters.

The modification of ver with a bridge grouping M¹ and / or M² linked camptothecin derivatives with carbohydrate residues can be different Methods and linker groups are done. Preferably z. B. the transfer of p- Amino-phenylglycosides in isothiocyanates and the linkage for example with amino groups. Carboxyalkyl or amino can also be used alkyl glycosides are easily coupled with amino or carboxyl groups.

Protective groups can be split off in appropriate reaction steps for example by exposure to acid or base, hydrogenolytically or on otherwise done reductively.  

Biological testing 1. Growth inhibition test to determine the cytotoxic characteristics

The human colon cell lines SW 480 and HT 29 (ATCC No. CCL 228 and HBT-38) and the mouse melanoma cell line B16F10 were in Roux dishes drawn in RPMI 1640 medium with the addition of 10% FCS. It was then trypsinized and made into one in RPMI plus 10% FCS Cell count of 50,000 cells / ml added. 100 µl cell suspension / well were placed in a 96 microwave plate and 1 day at 37 ° C in CO₂- Incubator incubated. Then another 100 ul RPMJ medium and 1 µl DMSO with the test substances added. The growth was checked after day 3 and day 6. For this purpose, 40 µl was added to each microwave MTT solution (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazoline bromide) added with an initial concentration of 5 mg / ml H₂O. It was 5th Incubated for hours in a CO₂ incubator at 37 ° C. Then that was Aspirated medium and 100 ul i-propanol / well added. After 30 min Shaking with 100 ul H₂O was the absorbance at 540 nm with a Titertek Multiskan MCC / 340 (Flow) measured.

The cytotoxic effect is in Table 1 as the IC₅₀ value for each SW 480 and HT 29 and B16F10 cell line specified:  

Table 1

2. Hematopoietic activity of glycoconjugates compared to underlying active ingredient material and methods

Bone marrow cells were rinsed from the mouse femur. 10⁵ cells were in McCoy 5A medium (0.3% agar) together with recombinant murine GM-CSF (Genzyme; stem cell colonization) and the substances (10 ^-4 to 100 µg / ml) at 37 ° C and 7 % CO₂ incubated. 7 days later, the colonies (<50 cells) and clusters (17-50 cells) were counted out.

Results

As shown in Table 2, the examined glycoconjugates show one compared to the underlying active ingredient drastically reduced hem Bone marrow stem cell proliferation.  

Table 2

Inhibition of CSF-induced proliferation of mouse bone marrow stem cells

3. In vivo inhibition of tumor growth in the nude mouse model material

For all in vivo experiments to investigate tumor inhibition athymic nude mice (NMRI nu / nu strain) were used for growth turns. The selected large cell lung carcinoma LXFL 529 was developed by serial passage in nude mice. The human primal Leap of the tumor was caused by isoenzymatic and immunohisto chemical methods documented.

Experimental setup

The tumor was subcutaneously in both flanks from 6 to 8 weeks old implanted nu / nu nude mice. Treatment became dependent on that Doubling time started as soon as the tumors had a diameter of Had reached 5-7 mm. The mice were the treatment group and the Control group (5 mice per group with 8-10 evaluable tumors) allocated by randomization. The individual tumors of the control group all grew progressively.

The size of the tumors was measured in two dimensions using a slide gauge measured. The tumor volume, which correlated well with the cell number, was then used for all evaluations. The volume was according to  using the formula "length × width × width / 2" ([a × b²] / 2, a and b stand for two diameters arranged at right angles).

The relative tumor volume (RTV) values were for each individual Tumor by dividing the tumor size on day X by the tumor size of day 0 (at the time of randomization). The middle ones RTV values were then used for further evaluation.

Inhibition of the increase in tumor volume (tumor volume of the test group / control group, T / C, in percent) was the final measurement.

treatment

The compounds were applied on days 1, 2 and 3 after Randomization intraperitoneally (i.p).

Results

The therapeutic activity is based on the compound from Example 1.1 the glycoconjugates according to the invention compared to the large cell human lung tumor xenograph LXFL 529. The therapy leads at the maximum tolerable dose (MTD) and at 1/2 MTD Tumor remission.  

Table 3

The compounds of the invention exhibit both in vitro and in vivo a surprisingly strong antitumor activity against various Tumors, especially those of the lungs and colon, associated with a great selectivity towards non-malignant cells.

They are therefore suitable for the treatment of cancer in particular those of the lungs and colon.  

The present invention includes pharmaceutical preparations which, in addition to non-toxic, inert pharmaceutically suitable excipients one or more contain more compounds according to the invention or those of one or more Active substances according to the invention exist, and methods for producing them Preparations.

The active ingredient (s) may optionally be present in one or more of the above specified carrier substances are also present in microencapsulated form.

The therapeutically active compounds are listed in the above pharmaceutical preparations preferably in a concentration of about 0.1 to 99.5, preferably from about 0.5 to 95% by weight of the total mixture be there.

The pharmaceutical preparations listed above can in addition to the inventions Compounds according to the invention also contain other active pharmaceutical ingredients.

In general it has been in both the human and the veterinary medicine has proven to be advantageous in the active ingredient (s) according to the invention Total amounts from about 0.5 to about 500, preferably 5 to 100 mg / kg body weight each 24 hours, if necessary in the form of several individual doses, to educate to deliver the desired results. A single dose contains the or the active compounds according to the invention preferably in amounts of about 1 to about 80, especially 3 to 30 mg / kg body weight.  

Examples Carbohydrate educts Example I.1 p-aminophenyl-3-O-methyl-β-L-fucopyranoside I.1.a) p-Nitrophenyl-3-O-methyl-β-L-fucopyranoside

6 g (21 mmol) p-nitrophenyl-β-L-fucopyranoside in 300 ml absolute. Methanol 7.84 g (31.5 mmol) of dibutyltin oxide are added and the mixture is refluxed for 2 h heated. It is then concentrated, the residue is dried and then in 300 ml of DMF added. After adding 15.7 ml of methyl iodide Batch stirred at 70 ° C for 40 h. The solvent is removed in vacuo and the residue was taken up in 300 ml of dichloromethane. The suspension becomes fil tried, the remaining solution concentrated again and a flash chromatography (Dichloromethane / methanol 99: 1). After concentration, 3.82 g are obtained (61%) of the target product.

I.1) p-aminophenyl-3-O-methyl-β-L-fucopyranoside

3.81 g (12.73 mmol) of p-nitrophenyl-3-O-methyl-β-L-fucopyranoside are dissolved in methanol and, after adding platinum dioxide, hydrogenated in a hydrogen atmosphere at a slight excess pressure. After filtering off the catalyst and precipitating with ether, 3 g (88%) of the target product are obtained. [TLC: dichloromethane / methanol 9: 1 R _f = 0.53].

Example I.2 p-aminophenyl-3-O-carboxymethyl-β-L-fucopyranoside I.2.a) p-Nitrophenyl-3-O-methoxycarbonylmethyl-β-L-fucopyranoside

1 g (3.5 mmol) of p-nitrophenyl-β-L-fucopyranoside and 1.3 g (5.2 mmol) of dibutyl Tin oxide are refluxed in 50 ml of methanol for 2 hours. The solution will be concentrated, the residue taken up in 50 ml of dioxane, with 2 ml of bromine methyl acetate and 100 mg of tetrabutylammonium iodide added and 16 h heated under reflux. The solvent is evaporated and the product through Flash chromatography (dichloromethane / methanol 99: 1) cleaned. After constriction the corresponding fractions and cases from methanol / ether give 455 mg (37%) of the target connection.

I.2) p-aminophenyl-3-O-carboxymethyl-β-L-fucopyranoside

282 mg (0.79 mmol) of p-nitrophenyl-3-methoxycarbonylmethyl-β-L-fucopyranoside are dissolved in 20 ml of methanol and mixed with 440 ul of an aqueous 2N lithium hydroxide solution. After stirring for 2 h at room temperature. is adjusted to pH 3 with an acidic ion exchanger SC108 and filtered. 250 mg of palladium on activated carbon are added to the filtrate. The mixture is then hydrogenated with hydrogen at a slight excess pressure for 1.5 h, the catalyst is separated off and washed with methanol. Concentration, absorption in water and freeze-drying lead to the target product (212 mg) in 86% yield. [TLC: aceto nitrile / water / glacial acetic acid 5: 1: 0.2 R _f = 0.24].

The following derivatives, for example, can be used as further carbohydrate building blocks:
p-aminophenyl-β-L-fucopyranoside
p-aminophenyl-2-O-methyl-β-L-fucopyranoside
p-aminophenyl-2-O-hydroxyethyl-β-L-fucopyranoside
p-aminophenyl-4-O-methyl-β-L-fucopyranoside
p-aminophenyl-3-O-methyl-α-L-fucopyranoside
p-aminophenyl-3-on-propyl-β-L-fucopyranoside
p-aminophenyl-3-deoxy-β-L-fucopyranoside
p-aminophenyl-3,4-dideoxy-β-L-fucopyranoside
p-aminophenyl-3,4-epoxy-β-L-fucopyranoside
p-aminophenyl-4-deoxy-β-L-fucopyranoside
p-aminophenyl-3-O-methoxycarbonylmethyl-β-L-fucopyranoside
p-aminophenyl-3-O-hydroxyethyl-β-L-fucopyranoside
p-aminophenyl-2-O-carboxymethyl-β-L-fucopyranoside
p-aminophenyl-3-O-succinyl-β-L-fucopyranoside
p-aminophenyl-3,4-di-O-methyl-β-L-fucopyranoside
p-aminophenyl -3-O-carbamoylmethyl-β-L-fucopyranoside
p-aminophenyl-α-L-rhamnopyranoside
p-aminophenyl-β-D-galactopyranoside
p-aminophenyl-2-O-methyl-β-D-galactopyranoside
p-aminophenyl-3-O-methyl-β-D-galactopyranoside
p-aminophenyl-4-O-methyl-βD-galactopyranoside
p-aminophenyl-6-O-methyl-β-D-galactopyranoside
p-aminophenyl-2,3-di-O-methyl-β-D-galactopyranoside
p-aminophenyl-2,4-di-O-methyl-β-D-galactopyranoside
p-aminophenyl-2,6-di-O-methyl-β-D-galactopyranoside
p-aminophenyl -3,4-di-O-methyl-β-D-galactopyranoside, acetate
p-aminophenyl-3,6-di-O-methyl-β-D-galactopyranoside
p-aminophenyl-4,6-di-O-methyl-β-D-galactopyranoside
p-aminophenyl-2,3,4-tri-O-methyl-β-D-galactopyranoside
p-aminophenyl-2,3,6-tri-O-methyl-β-D-galactopyranoside
p-aminophenyl2,4,6-tri-O-methyl-β-D-galactopyranoside
p-aminophenyl-3,4,6-tri-O-methyl-β-D-galactopyranoside
p-aminophenyl-3-deoxy-β-D-galactopyranoside
p-aminophenyl-3,4-dideoxy-β-D-galactopyranoside
p-aminophenyl-6-O-acetyl-β-D-galactopyranosi d
p-aminophenyl-3-O-methoxycarbonylmethyl-β-D-galactopyranoside
p-aminophenyl-3-O-carboxymethyl-β-D-galactopyranoside, sodium salt
p-aminophenyl-3-O-carbamoylmethyl-β-D-galactopyranoside
p-aminophenyl-3-O- (N-methyl-carbamoylmethyl) -β-D-galactopyranoside
p-aminophenyl-α-D-mannopyranoside
p-aminophenyl-3-O-methyl-α-D-mannopyranoside
p-aminophenyl-2,3-di-O-methyl-α-D-mannopyranoside
p-aminophenyl-3-O-methoxycarbonylmethyl-α-D-mannopyranoside
p-aminophenyl-3-O-carboxymethyl-α-D-mannopyranoside
p-aminophenyl-3-O-carbamoylmethyl-α-D-mannopyranoside
p-aminophenyl-4-O- (β-D-galactopyranosyl) -β-D-glucopyranoside
p-Aminophenyl-4-O- (3'-sulfate-β-D-galactopyranosyl) -β-D-glucopyrano sid, sodium salt
p-Aminophenyl-4-O- (3'-O-methyl-β-D-galactopyranosyl) -β-D-glucopyranoside
p-Aminophenyl-2-O-methyl-4-O- (3'-O-methyl-β-D-galactopyranosyl) -β-D-gluco pyranoside
p-Aminophenyl-4-O- (3 ', 4'-di-O-methyl-β-D-galactopyranosyl) -β-D-glucopyranoside.

The synthesis of the following carbohydrate building blocks was already described in EP 501 250:
Carboxymethyl-β-L-fucopyranoside
5-carboxypentyl-β-L-fucopyranoside.

The linkage of these two building blocks with peptide conjugates can be found in the EP 501 250 described manner.  

Glycoconjugates Example 1.1 20 (S) -7-ethyl-20-O- {N ^α , N ^ε -bis- [O- (3-O-methyl-β-L-fucopyranosy) -4-hydroxy-phenylamino-thiocarbonyl] -lysyl- alanyl} -camptothecin 1.1.a) 20 (S) -7-ethyl-20-O- [N- (tert-butoxycarbonyl) alanyl] camptothecin

A solution of 1.88 g (5.0 mmol) of 20 (S) -7-ethyl camptothecin (S. Sawada et al., Chem. Pharm. Bull., 39 (1991) 1446-1454) in 100 ml of absolute Dimethylformamide is mixed with stirring with 2.15 g (10.0 mmol) of N- (tert-butoxycarbonyl) -alanine-N-car bonic anhydride and 150 mg (1.2 mmol) of 4- (N, N-dimethylamino) pyridine . After 3 h at room temperature, a further 2.15 g (10.0 mmol) of N- (tert-butoxycarbonyl) alanine-N-carboxylic acid anhydride and 150 mg (1.2 mmol) of 4- (N, N-dimethylamino) - pyridine and stir overnight at room temperature. The mixture is then concentrated in vacuo and the residue is purified by flash chromatography [petroleum ether / ethyl acetate 2: 1 → 1: 1 → ethyl acetate]. Colorless crystals (2.02 g, 73.8%) are obtained; TLC [ethyl acetate]: R _f = 0.56; Mp = 206-212 ° C; FAB-MS: m / z = 548 (M + H⁺).

1.1.b) 20 (S) -20-O-Alanyl-7-ethyl-camptothecin, trifluoroacetate

A solution of compound 1.1.a (1.81 g, 3.3 mmol) in a mixture of 70 ml dichloromethane and 7 ml anhydrous trifluoroacetic acid is for 90 min. stirred at room temperature. After concentration in vacuo to a small volume, the product is precipitated with diethyl ether and washed thoroughly with diethyl ether. The product is obtained in light yellow crystals (1.34 g, 72.3%); TLC [ethyl acetate]: R _f = 0.05; Mp = 242 ° C (decomp.).

1.1.c) 20 (S) -7-ethyl-20-O- [N ^α , N ^ε -di- (tert-hutoxycarbonyl) -ysyl-alanyl] - camptothecin

1.57 g (4.55 mmol) of N, N-di- (tert-butoxycarbonyl) -lysine and 923 mg (6.83 mmol) of 1-hydroxy-1H-benzotriazole hydrate are dissolved in 35 ml of dimethylformamide. After adding 1.09 g (5.7 mmol) of N-ethyl-N '- (dimethylaminopropyl) carbodiimide hydrochloride and 990 μl (5.7 mmol) of ethyl-diisopropylamine, the mixture is stirred for 30 min. at room temperature. A solution of compound 1.1.b (1.3 g, 2.32 mmol) in 35 ml of dimethylformamide and 408 μl (2.32 mmol) of ethyl-diisopropylamine is then added and the mixture is stirred for a further 16 h at room temperature. After concentration in vacuo and purification by flash chromatography [petroleum ether / ethyl acetate 2: 1 → 1: 1 → ethyl acetate], pale yellow crystals are obtained (1.38 g, 75.3%); TLC [ethyl acetate]: R _f = 0.53; Mp = 125 ° C (dec.).

1.1.d) 20 (S) -7-ethyl-20-O- (Iysyl-alanyl) -camptothecin, di-trifluoroacetate

An anhydrous trifluoroacetic acid (5 ml) is added to a suspension of the above compound (1.18 g, 1.5 mmol) in dichloromethane (50 ml) and the resulting solution is stirred for 1 h at room temperature. After concentration to a small volume in vacuo, the product is precipitated by adding diethyl ether. The precipitate is filtered off and recrystallized from ethyl acetate. Yellow crystals are obtained (862 mg, 71.5%); TLC [ethyl acetate]: R _f = 0.05; Mp = 137 ° C (dec.).

1.1) 20 (S) -7-ethyl-20-O- {N ^a , N ^ε -bis- [O- (3-O-methyl-βfucopyranosyl) -4-hydroxy-phenylamino-thiocarbonyl] -lysyl-alanyl} -camptothecin

A solution of 269.4 mg (1.0 mmol) of p-aminophenyl-3-O-methyl-β-L-fuco pyranoside (Example I.1) in dioxane / water 1: 1 (10 ml) is stirred with thio phosgene (150 µl, 2.0 mmol) was added. After 10 min. add 4 equiva lenten ethyldiisopropylamine, then concentrated in vacuo and dries the Residue for 1 h in an oil pump vacuum. The isothiocyanate obtained is in 50 ml of absolute dimethylformamide dissolved and with 361.7 mg (0.45 mmol) of Compound 1.1.d and 317 ul (1.8 mmol) of ethyl diisopropylamine were added. Man Stir for 16 h at room temperature, then concentrate in vacuo and clean through Flash chromatography [acetonitrile / water 15: 1 → 10: 1 → 5: 1]. The product will reprecipitated from dichloromethane / methanol with diethyl ether. 203.3 mg are obtained (37.7%) of the target product as yellow crystals; Mp = 196-198 ° C (dec.).

Example 1.2 1.1) 20 (S) -7-ethyl-20-O- {N ^α , N ^ε -bis- [O- (3-O-carboxymethyl-β-L-fucopyranosyl) -4-hydroxy-phenylamino-thiocarbonyl] - lysyl-alanyl} -camptothecin, disodium salt

Analogously to Example 1.1, the glycoconjugate is made from 361.7 mg (0.45 mmol) of peptide conjugate 1.1.d and 313.3 mg (1.0 mmol) of p-aminophenyl-3-O-carboxy methyl-β-L-fucopyranoside (Example I.2). Purification by flash chromatography [acetonitrile / water 10: 1 → 5: 1]. Conversion to the disodium salt with 2 equivalents of a 0.1N sodium hydroxide solution. A yellow, amorphous solid is obtained (162.8 mg, 27.2%); TLC [aceto nitrile / water 5: 1]: R _f = 0.09.

Example 1.3 20 (S) -7-ethyl-20-O- {N ^α - [O- (3-O-carboxymethyl-β-L-fucopyranosy) -4-hydroxy-phenylamino-thiocarbonyl] -syl-alanyl} -camptothecin, Hydrochloride 1.3.a) 20 (S) -7-ethyl-20-O- [N ^ε - (fluorenyl-9-methoxycarbonyl) -Iysyl-alanyl] - camptothecin, trifluoroacetate

The conjugate from Example 1.1.b is prepared in analogy to the instructions from Example 1.1.c. linked with N ^α - (tert-butoxycarbonyl) -N ^ε - (fluorenyl-9-methoxycarbonyl) -lysine and then to the α-amino function according to Example 1.1.d. unblocked.

1.3.b) 20 (S) -7-ethyl-20-O- {N ^α - [O- (3-O-carboxymethyl-β-L-fucopyranosyl) -4-hydroxy-phenylamino-thiocarbonyl] -N ^ε - [fluorenyl-9-methoxycarbonyl] lysyl alanyl} camptothecin

The compound from Example 1.3.a is analogous to Example 1.1 with 1.2 equi valents of the carbohydrate derivative from Example I.2 implemented.  

1.3) 20 (S) -7-Ethyl-20-O- {N ^a - [O- (3-O-carboxymethyl-β-L-fucopyranosyl) -4-hydroxy-phenylamino-thiocarbonyl] -lysyl-alanyl} - camptothecin, hydrochloride

The conjugate 1.3.b is deblocked with piperidine in DMF. After 30 min. you narrow and digest the residue twice with dichloromethane. Then in DMF taken up and precipitated with methanol / ether. The product is suctioned off with Washed ether and then lyophilized from dioxane / water. Then takes one in water; mixed with 1 equivalent of a 0.01 N HCl solution and lyophi again.

Example 1.4 20 (S) -7-ethyl-20-O- (N ^α - [O- (3-O-carboxymethyl-β-L-fucopyranosyl) -4-hydroxy-phenylamino-thiocarbonyl] -lysyl-valyl} -camptothecin, Hydrochloride

The connection is established analogously to 1.3.  

Example 2.1 20 (R, S) -10,11-methylenedioxy-20-O- {N ^α , N ^ε -bis- [O- (3-O-methyl-β-L-fuco pyranosyl) -4-hydroxy-phenylamino- thiocabonyl] lysyl alanyl} camptothecin 12.1.a) 20 (B / S) -10,11-methylenedioxy-camptothecin

This camptothecin derivative is according to Wall et. al. (J. Med. Chem., 29 (1986), 2358) from the racemic tricyclic.

2.1.b) 20 (R, S) -10,11-methylenedioxy-20-O-alanyl-camptothecin, trifluoroacetate

A solution of 300 mg (0.765 mmol) 20 (R / S) -10,11-methylenedioxy-campto thecin in 30 ml absolute dimethylformamide is stirred with 330 mg (1.53 mmol) N- (tert-butoxycarbonyl) -alanine -N-carboxylic acid anhydride and 30 mg of 4- (N, N-dimethylamino) pyridine were added. After stirring for 24 h at room temperature, two further equivalents of N- (tert-butoxycarbonyl) alanine-N-carboxylic anhydride and 30 mg of 4- (N, N-dimethylamino) pyridine are added. After the reaction has ended, the mixture is concentrated in vacuo. After precipitation from dichloromethane with ether, the protected intermediate is obtained in 80% yield. [TLC: dichloromethane / methanol 95: 5 R _f = 0.38]. It is taken up in 10 ml of dichloromethane and 2 ml of anhydrous trifluoroacetic acid are added at 0 ° C. After 15 min. is concentrated and precipitated from dichloromethane / methanol with ether.
Yield: 97% [FAB-MS: m / z = 464 = M + H].

2.1) 20 (R, S) -10,11-methylenedioxy-20-O- {N ^α , N ^ε -bis- [O- (3-O-methyl-β-L-fucopyranosyl) -4-hydroxy-phenylamino -thiocarbonyl] -lysyl-alanyl} - camptothecin

The glycoconjugate is prepared in analogy to the instructions for 1.1.c, 1.1.d and 1.1 from the alanyl conjugate in Example 2.1.b.
Yield: 87% [TLC: acetonitrile / water 10: 1 R _f = 0.33] [FAB-MS: m / z = 1214 = M + H].

Example 2.2 20 (S) -10,11-methylenedioxy-20-O- {N ^α , N ^ε -bis- [O- (3-O-methyl-β-L-fucopyranosyl) -4-hydroxy-phenylamino-thiocarbonyl] - lysyl-alanyl} - camptothecin 2.2.a) 20 (S) -10,11-methylenedioxy camptothecin

This camptothecin derivative is according to Wall et. al. (J. Med. Chem., 29 (1986), 2358) from the S-configured, enantiomerically pure tricyclic compound, which is characterized by Racemate resolution can be obtained.

2.2) 20 (S) -10,11-Methylenedioxy-20-O- {N ^α , N ^ε -bis- [O- (3-O-methyl-β-L-fucopyranosyl) -4-hydroxy-phenylamino-thiocarbonyl ] -lysyl-alanyl} - camptothecin

Representation in analogy to example 2.1.  

Example 2.3 20 (S) -10,11-methylenedioxy-20-O- {N ^a - [O- (3-O-carboxymethyl-β-L-fuco pyranosyl) -4-hydroxy-phenylamino-thiocarbonyl] -lysyl-alanyl} -camptothecin, hydrochloride 2.3.a) 20 (S) -10,11-methylenedioxy-20-O-alanyl-camptothecin, trifluoroacetate

This conjugate is produced from compound 2.2.a in analogy to 2.1.b.
Education: 72% over 2 stages [FAB-MS: m / z = 464 = M + H].

2.3.b) 20 (S) -10,11-Methylenedioxy-20-O- {1N ^ε - [fluorenyl-9-methoxycarbonyl] - lysyl-alanyl} -camptothecin, trifluoroacetate

The conjugate from Example 2.3.a is prepared in analogy to the instructions from Example 1.1.c. linked with N ^α - (tert-butoxycarbonyl) -N ^ε - (fluorenyl-9-methoxycarbonyl) -lysine and then deblocked at the α-amino function by the action of trifluoroacetic acid. Education: 52% over 2 levels.

2.3.c) 20 (S) -10,11-Methylenedioxy-20-O- (N ^α [O- (3-O-carboxymethyl-β-L-fuco pyranosyl) -4-hydroxy-phenylamino-thiocarbonyl] -N ^ε - [fluorenyl-9-methoxycarbonyl] lysyl alanyl} camptothecin

The compound from Example 2.3.b is reacted in analogy to Example 1.1 with 1.2 equivalents of the carbohydrate derivative from Example I.2. The crude product is purified by stirring with water and subsequent precipitation from dichloromethane / methanol with ether.
Yield: 88% [TLC: dichloromethane / methanol / ammonia 17% 15: 4: 0.5 R _f = 0.27]

2.3.) 20 (S) -10,11-Methylenedioxy-20-O- {N ^α [O- (3-O-carboxymethyl-β-L-fucopyranosyl) -4-hydroxy-phenylamino-thiocarbonyl] -lysyl-alanyl } - camptothecin, hydrochloride

The conjugate 2.3.c is deblocked with piperidine in DMF. After 20 min. The mixture is concentrated and the residue is digested twice with dichloromethane. The product is filtered off, washed again with ether and then lyophilized from dioxane / water. Then take up in dioxane / water; mixed with 1 equivalent of a 0.1 N HCl solution and lyophilized again. Education: quant. [DC: aceto nitrile / water / glacial acetic acid 5: 1: 0.2 R _f = 0.26] [FAB-MS: m / z = 947 = M + H]

Example 3.1 20 (S) -9-amino-20-O- {N ^α -N ^ε -bis- [O- (3-O-methyl-β-L-fucopyranosyl) -4-hydroxy-phenylamino-thiocarbonyl] -lysyl- alanyl} -camptothecin 3.1.a) 20 (S) -9-nitro-camptothecin

This derivative is according to the instructions of Wani et al. (J. Med. Chem. 29 (1986), 2358).

3.1.b) 20 (S) -9-nitro-20-O-alanyl camptothecin, trifluoroacetate

516 mg (1.31 mmol) of the compound from Example 3.1.a are stirred in 55 ml of absolute dimethylformamide with 846 mg (3.9 mmol) of N- (tert-butoxycarbonyl) alanine-N-carboxylic anhydride and 55 mg of 4- (N, N-Dimethylamino) pyridine added. After 4 h, the mixture is concentrated in vacuo and chromatographed on silica gel using dichloromethane / ethyl acetate 2: 1. After using the recovered starting material again under the same conditions, the protected intermediate is obtained in 54% yield. [TLC: dichloromethane / methanol 95: 5 R _f = 0.5]. It is taken up in 40 ml of dichloromethane and 4 ml of anhydrous trifluoroacetic acid are added at 0 ° C.

After 15 min. is concentrated and purified by stirring with ether. Education: 80%

3.1.c) 20 (S) -9-amino-20-O-alanyl-camptothecin, trifluoroacetate

320 mg (0.55 mmol) of the compound from 3.1.b are applied in 30 ml of ethanol take and 15 min. hydrogenated over 10 mg of platinum dioxide. The catalyst is off filtered, the solution concentrated and the product after precipitation from dichloromethane used with methanol in the next stage.

3.1.d) 20 (S) -9-amino-20-O- (lysyl-alanyl) -camptothecin, di-trifluoroacetate

31 mg (0.09 mmol) of N, N-di- (tert-butoxycarbonyl) lysine are dissolved in 5 ml of dimethylformamide with 15 mg (0.135 mmol) of N-hydroxysuccinimide and 22 mg (0.11 mmol) of dicyciohexyl preactivated carbodiimide for 2 h. Then compound 3.1.c (60 mg, 0.09 mmol) and 31 μl of ethyl-diisopropylamine are added and the mixture is stirred for a further 16 h at room temperature. After concentration in vacuo and purification by flash chromatography [toluene / ethanol 8: 1], the intermediate is obtained in a yield of 47%. [TLC: dichloromethane / methanol 9: 1 R _f = 0.31]. 30 mg (0.039 mmol) in dichloromethane (10 ml) are mixed with anhydrous trifluoroacetic acid (2 ml) and the resulting solution for 30 min. stirred at room temperature. After concentration, the product is precipitated from dichloromethane / methanol with ether (20 mg, 57%). [TLC: acetonitrile / water / glacial acetic acid 10: 5: 3 R _f = 0.44].

3.1) 20 (S) -9-amino-20-O- {N ^α , N ^ε -bis- [O- (3-O-methyl-β-L-fucopyranosyl) -4-hydroxy-phenylamino-thiocarbonyl] - lysyl-alanyl} -camptothecin

The glycoconjugate is prepared from the compounds in 3.1.d and I.1 in analogy to Example 1.1. Yield: 58% [TLC: acetonitrile / water / glacial acetic acid 5: 1: 0.2 R _f = 0.6].

Example 4.1 20 (S) -7- {N ^α , N ^ε -bis- [O- (3-O-methyl-β-L-fucopyranosyl) -4-hydroxy-phenylamino-thiocarbonyl] -lysyl-alanyloxymethyl} -20-O - {N ^α , N ^ε -bis- [O- (3-O-methyl-β-L-fucopyranosyl) -4-hydroxy-phenylamino-thiocarbonyl] -lysyl-alanyl} - camptothecin 4.1.a) 20 (S) -7-hydroxymethyl camptothecin

This connection is carried out according to the instructions of Miyasaka et al. (Chem. Pharm. Bull. 39 (1991) 2574).

4.1.b) 20 (S) -7- (Alanyloxymethyl) -20-O-alanyl-camptothecin, di-trifluoroacetate

1 g (2.64 mmol) of 20 (S) -7-hydroxymethyl-camptothecin are dissolved in 100 ml of DMF and then with 100 mg of 4-N, N-dimethylaminopyridine and 1.5 equivalents of N- (tert-butoxycarbonyl) - L-alanine-N-carboxy-anhydride are added and the suspension is stirred at room temperature for 16 h. To achieve complete double acylation, 100 mg of 4-N, N-dimethylaminopyridine and 1.5 equivalents of N- (tert-butoxycarbonyl) alanine-N-carboxy-anhydride are again added and the mixture is left at room temperature for a further 16 h. It is concentrated and purified by flash chromatography on ethyl acetate / petroleum ether 1: 1. The purified material is taken up in 30 ml of dichloromethane and 5 ml of trifluoroacetic acid are added at 0 ° C. After stirring for 30 min, the mixture is concentrated and the amino-blocked product is precipitated from dichloromethane / ether. The mixture is then lyophilized from dioxane / water. [TLC: acetonitrile / water / glacial acetic acid 10: 5: 5 R _f = 0.17] [FAB-MS: m / z = 777 = M + H].

4.1) 20 (S) -7- {N ^α , N ^ε -bis- [O- (3-O-methyl-β-L-fucopyranosyl) -4-hydroxy-phenylamino-thiocarbonyl] -lysyl-alanyloxymethyl} -20 -O- {N ^α , N ^ε -bis- [O- (3-O-methyl-β-L-fucopyranosyl) -4-hydroxy-phenylamino-thiocarbonyl] - lysyl-alanyl} -camptothecin

The conjugate is prepared from Example 4.1.b in analogy to Examples 1.1.c, 1.1.d and 1.1, but at all stages with twice the amount of the respective reactants. [TLC: acetonitrile / water 10: 1 R _f = 0.2] [MALDI-MS: m / z = 2047 = M (+3) + Na].

Claims (10)

1. Compounds of the general formula (I) A-Cp-B (I) in which
Cp for a group of formulas wherein
R¹, R², R³ and R⁴ are independently hydrogen; Alkyl with up to 3 carbon atoms, halogen, amino, hydroxy or nitro can mean or
R² and R³ together form a group of the formula mean where
m can have the values 1 or 2, and
R⁵ for H, -CH₂CH₃, -CH₂-O- *, -CH₂-NH *, stands for or -CH₂-N (CH₂CH₃) R⁶ or stands,
wherein R⁶ means arylmethyl or hetarylmethyl, wherein
R⁷ and R⁸ are defined as R² and R³ and can be the same or different with them, wherein
R⁹ represents hydrogen or -CH₂-N (CH₃) ₂ and
R¹⁰ represents hydrogen or ethyl,
or wherein
R¹¹ and R¹² are defined as R² and R³ and may be the same or different with them, or wherein
R¹³ and R¹⁴ are defined as R² and R³ and can be the same or different with them,
where Cp is linked to A at the positions marked with # and B to the positions marked with *,
A is a residue of the formula means, where 1 (n + o + p) 3 applies,
B is hydrogen or a radical of the formula means, where 0 (q + r + s) 3, where
M¹ and M² independently represent a bridging group whose main chain comprises up to 21 atoms in a linear sequence,
L¹, L², L³, L⁴, L⁵ and L⁶ independently of one another stand for linker groups, as are commonly used in glycoconjugate chemistry,
Sp¹, Sp², Sp³, Sp⁴, Sp⁵ and Sp⁶ independently represent arylene with up to 10 carbon atoms or for alkylene with up to 8 carbon atoms, which are each optionally substituted, and
K¹, K², K³, K⁴, K⁵ and K⁶ independently of one another for a radical of the formula (II) stand in what
E is methyl, hydroxymethyl, alkoxymethyl having up to 6 carbon atoms, acyloxymethyl having up to 6 carbon atoms or a radical of the formula -CH₂-D, in which
D represents a radical of the formula (II),
R¹⁵, R¹⁶ and R¹⁷ are independently hydrogen; Hydroxy, optionally substituted by hydroxyl-substituted alkoxy having up to 6 carbon atoms, optionally substituted by alkyl or acyl having up to 6 carbon atoms, amino, halogen, sulfate or a group of the formulas mean what
R¹⁸ and R¹⁹ independently of one another represent hydroxyl or alkoxy having up to 6 carbon atoms or amino, which is optionally substituted by alkyl having up to 6 carbon atoms, and
u and v can independently assume the values 0, 1, 2, 3 or 4
or
R¹⁵, R¹⁶ and R¹⁷ independently represent a radical of the formula (II)
or
two of the radicals R¹⁵, R¹⁶, R¹⁷ together represent an epoxy group,
and their isomers, mixtures of isomers and salts. 2. Compounds of general formula (I) according to claim 1, wherein K¹, K², K³, K⁴, K⁵ and K⁶ can independently of one another represent a radical of formula (II), wherein
E denotes methyl, hydroxymethyl, methoxymethyl or acetoxymethyl,
R¹⁵ is hydrogen; Hydroxyl, methoxy or a group of the formulas means what
u and v can independently assume the values 1 or 2 and
R¹⁸ and R¹⁹ independently of one another represent hydroxyl or alkoxy having up to 4 carbon atoms,
or
R¹⁵ represents a radical of the formula (II),
R¹⁶ is hydrogen, hydroxyl, halogen, alkoxy with up to 4 carbon atoms, sulfate or a group of the formulas means what
u and v can independently assume the values 1 or 2 and
R¹⁸ and R¹⁹ independently of one another represent hydroxy or alkoxy having up to 4 carbon atoms or amino, which is optionally substituted by alkyl having up to 4 carbon atoms,
R¹⁷ hydroxyl, alkoxy with up to 4 carbon atoms, which is optionally substituted by hydroxy, optionally substituted by alkyl or acyl with up to 4 carbon atoms, or a group of the formulas means what
u and v can independently assume the values 1 or 2 and
R¹⁸ and R¹⁹ independently of one another represent hydroxyl or alkoxy having up to 4 carbon atoms,
or in what
R¹⁵ and R¹⁶ together represent an epoxy group,
and their isomers, mixtures of isomers and salts. 3. Compounds of general formula (I) according to claim I, wherein Sp¹, Sp², Sp³, Sp⁴, Sp⁵ and Sp⁶ independently of one another for aryls with up to Can stand 10 carbon atoms, each with a group K¹, K², K³, K⁴, K⁵ or K⁶ and L¹, L², L³, L⁴, L⁵ or L⁶ is linked and that optionally one or more times by hydroxy, carboxy, Carboxyalkyl with up to 4 carbon atoms, nitro, cyano, halogen, Alkyl with up to 4 carbon atoms, haloalkyl with up to 4 Carbon atoms or by alkoxy with up to 4 carbon atoms is substituted, as well as their isomers, isomer mixtures and salts. 4. Compounds of general formula (I) according to claim 1, wherein L¹, L², L³, L⁴, L⁵ and L⁶ independently of one another or mean where
R²⁰ represents chlorine or hydroxyalkylamino with up to 6 carbon atoms,
and their isomers, mixtures of isomers and salts. 5. Compounds of general formula (I) according to claim 1, wherein M¹ and M² can independently represent a peptide which via an amino function is linked to L¹, L², L³, L⁴, L⁵ and / or L⁶, via an acyl function is linked to Cp and its amino acid building blocks can optionally carry protective groups and their isomers, Mixtures of isomers and salts. 6. Compounds according to claim 5, characterized in that M₁ and / or M₂ is a mono-, di- or tripeptide. 7. Compounds according to claim 5 or 6, characterized in that the Peptide from amino acid units which may have protective groups is selected from the group consisting of glycyl, alanyl, valyl, Leucyl, lysyl, seryl, glutamyl, threonyl, asparagyl, isoleucyl, Diaminopropionyl, diaminobutyryl, histidyl and / or ornithyl. 8. A process for the preparation of compounds of the general formula (I) according to claim 1, characterized in that compounds of the general formula (III) H _B -Cp-H _A (III) wherein Cp has the meaning given above and the hydrogen atom H _A at the position marked with # and H _B at the position marked with *, optionally after replacing H _B with a protective group, with an activated carboxyl component M¹a corresponding to the radical M¹ defined above, which optionally carries protective groups, in a suitable solvent, optionally in In the presence of a base, reacted by customary methods, optionally by means of known methods, one, more or all of the protective groups of M¹ are split off and the product is reacted with compounds of the general formula (IV) K¹-Sp¹-L¹a (IV) wherein K¹ and Sp¹ are as defined above and L¹a stands for a reactive precursor of the group L¹, reacted, the protective groups optionally being split off selectively and gradually shifting Different groups K²-Sp²-L²- and K³-Sp³-L³- can be introduced in a comparable manner, and that, if a carbohydrate component is to be linked to the position marked with *, the protective group, which replaces H _B , may be selectively selected , split off using known methods, and in the manner described above introduces the residue M² and, as desired, residues of the formulas K⁴-Sp⁴-L⁴-, K⁵-Sp⁵-L⁵- and K⁶-Sp⁶-L⁶-
or that, if M 1 and / or M 2 is a peptide, a first amino acid residue in the form of the corresponding carboxyl component, optionally bearing protective groups, is introduced in a comparable manner by conventional methods, if necessary splitting off protective groups, furthermore optionally linking amino acid residues bearing protective groups, as indicated above Introduces formulas K¹-Sp¹-L¹-, K²-Sp²-L²-, K³-Sp³-L³-, K⁴-Sp⁴-L⁴-, K⁵-Sp⁵-L⁵- and / or K⁶-Sp⁶-L⁶- and if necessary remove protective groups splits,
and that the stereo isomers are separated, if appropriate by customary methods, and that the compounds are optionally converted into their salts. 9. Use of the compounds of general formula (I) according to Claim 1 for the manufacture of pharmaceuticals. 10. Medicament containing compounds of the general formula (I) according to Claim 1.