WO1998023626A1

WO1998023626A1 - Antiadhesive benzoic acid derivatives

Info

Publication number: WO1998023626A1
Application number: PCT/EP1997/006308
Authority: WO
Inventors: Gerhard Kretzschmar; Alexander Toepfer
Original assignee: Glycorex Ab
Priority date: 1996-11-25
Filing date: 1997-11-12
Publication date: 1998-06-04
Also published as: DE19648681A1

Abstract

The invention relates to benzoic acid derivatives, in which α-C-fucosides or α-C-mannosides and alkyl chains or rings, which have at least one acid function, are attached in positions 3 and 5 of the aromatic ring, via ether linkages and short spacer groups. The invention relates furthermore to a procedure for the production of these compounds and their application in the production of medicaments and diagnostic reagents.

Description

description

Anti-adhesive benzoic acid derivatives

The invention relates to derivatives of benzoic acid in which α-C-fucosides or α-C-mannosides and chains or rings which carry at least one acid function have been attached in the 3- and 5-positions of the aromatic ring via ether bonds and short spacer groups. The invention also relates to the preparation of these compounds and their use in the manufacture of medicaments and diagnostics.

The circulation of blood cells such as Leukocytes, neutrophies, granulocytes and monocytes are a multi-stage, very complex process at the molecular level, which is only known in part steps (Review: T.A. Springer, Cell 76, 301-314, 1994).

Recent research results have shown that the crucial recirculation of lymphocytes in immune surveillance and the localization of neutrophils and monocytes in inflammation foci obey very similar molecular mechanisms. In acute and chronic inflammatory processes, leukocytes adhere to endothelial cells and migrate to the focus of inflammation and the secondary lymphoid organs.

Numerous specific signaling molecules such as interleukins, leukotrienes and tumor necrosis factor (TNF), their G-protein coupled receptors and in particular tissue-specific cell adhesion molecules are involved in this process, which ensure a precisely controlled recognition of the immune and endothelial cells. The most important adhesion molecules involved, which will be referred to as receptors in the following, include the selectins (E, P and L selectins), integrins and the members of the immunoglobulin superfamily. The three selectin receptors determine the initial phase of leukocyte adhesion. E-selectin is applied to endothelial cells a few hours after stimulation, for example as expressed by interleukin-1 (IL-1β) or tumor necrosis factor (TNF-α), while P-selectin is stored in platelets and endothelial cells and, after stimulation by thrombin, peroxide radicals or substance P, is presented on the cell surfaces, among other things. L-selectin is permanently expressed on leukocytes, but is quickly cleaved from the leukocytes again in the course of the inflammation.

The adhesion of leukocytes to endothelial cells mediated by selectin receptors in the initial phase of inflammatory processes is a natural and necessary immune response to various inflammatory stimuli and injuries to the vascular tissue. However, the course of a number of acute and chronic diseases is adversely affected by the excessive adhesion of leukocytes and their infiltration into the affected tissue and by the damage to healthy tissue in the sense of an autoimmune reaction. These include, for example, rheumatism, reperfusion injuries such as myocardial ischemia / infarction (MI), acute pneumonia after surgery, traumatic shock and stroke, psoriasis, dermatitis, ARDS (respiratory distress syndrome in adults) as well as after surgery (e.g. angioplasty and by-pass operations) occurring restenosis.

The natural ligand with the structure of the complex oligosaccharide Sialyl LewisX (SLeX) (Formula Scheme 1) has already been successfully used in animal experiments with P-selectin-dependent lung injuries (MSMulligan et al., Nature 1993, 364, 149) and with myocardial reperfusion injuries (M.Buerke et al., J.Clin.Invest. 1994, 93, 1140). However, in the first clinical trials in myocaric infarction, doses of several grams of this active ingredient had to be administered, since it has only a very low bioavailability (communication from Cytel Corp, / La Jolla, CA., published in Scrip No 2137, June 14, 1996, page 22). This high dose of active ingredient is also in line with the known weak affinity of the natural SLeX / A ligands for the selectin receptors in static in vitro test systems: SLeX inhibits the cell in all known in vitro test systems. adhesion to selectin receptors only at a relatively high concentration in the range of IC ₅₀ about 1 mM or above (Jacob et al., Biochemistry 1995, 34, 1210).

Formula scheme 1: Sialyl Lewis ^x

Efforts have been reported in numerous publications to achieve more firmly binding antagonists by structural variation of the SLeX ligand (see e.g. Review in Drugs of the Future 1995, 20, 805-816). The aim of this work is to provide more effective inhibitors of leukocyte adhesion in vivo. A comparable or even higher specific affinity for the selectin receptors and at the same time a significant structural simplification of an antagonist with the potential for improved bioavailability (also orally) has not yet been achieved. Thus, numerous so-called SLeX mimetics are actually still complex derivatives with the oligosaccharide structure (examples in J.Med Chem. 1996, 39, 1339-1343), or the glycosidic bonds were broken down by the even more labile serum peptide bonds. replacements (examples in Bioorgan. Med. Chem. Lett. 1996, 4, 1149-1165).

The object of the present invention is to produce new selectin ligands which are much easier to prepare than SLeX itself and which contain no glycosidic or peptide structural features. These substances therefore have an inherently higher bioavailability compared to the natural ligands with the SLeX structure or with peptide structural elements and would be much easier and cheaper to produce. Surprisingly, it was found that simple benzoic acid derivatives act as antagonists of the selectins and inhibitors of leukocyte adhesion and apparently contain all the minimally required structural features of the complex SLeX ligand.

Accordingly, the object is achieved by a compound of formula I.

in what mean

X oxygen, sulfur or 2 hydrogen atoms,

Y OH, NH ₂ , NHCH ₂ COOH, NHCH (COOH) ₂ , NHCH ₂ CH ₂ OH,

NHCH ₂ CH ₂ CH ₂ OH, NHCH ₂ CH ₂ CH ₂ CH ₂ OH,

R ¹ C-glycosidically linked fucose or mannose,

R ² (CH ₂ ) _π COOH, (CH ₂ ) _n SO ₃ H, (CH ₂ ) _n CH (COOH) ₂ , CH (COOH) ₂ , or COR ³ , where n is an integer from 1 to 5 and

R ^{3 is} an amino acid residue of the formulas NH (CH ₂ ) _m COOH, NH (CH ₂ ) _m SO ₃ H, or NHCH (R) COOH, pyrrolidine-2-carboxylic acid, pyrrolidine-3-carboxylic acid, piperidine-2-carboxylic acid, piperidine -3-carboxylic acid or piperidine-4-carboxylic acid and where m is an integer from 1 to 5 and

R ^{4 represents} an amino acid side chain of an amino acid selected from the group of naturally occurring proteinogenic α-amino acids. The amino acid side chain of glycine is, for example, -H, that of alanine, for example, -CH ₃ , that of valine, for example, -CH (CH ₃ ) _2, and the amino acid side chain of phenylalanine, for example, -CH ₂ -C ₆ H ₅ . The radical R ¹ in formula I is preferably

The following are examples of compounds with the preferred properties mentioned:

1 . A compound of formula 1 a, which is characterized in that

X oxygen,

Y OH,

R ¹ α1- (CH ₂ ) ₃ -glycosidically linked L-fucose,

R ^{2 means} (CH ₂ ) _n COOH and n = 1.

1 a 2. A compound of formula 1 b, which is characterized in that

X oxygen,

Y OH,

R ¹ α1- (CH ₂ ) ₃ -glycosidically linked L-fucose,

R ² COR ³ , where R ^{3 represents} a 1- (4-carboxy) piperidyl radical.

1b

3. A compound of formula 1 c, which is characterized in that X is oxygen,

Y OH,

R ¹ denotes α1- (CH ₂ ) ₃ -glycosidically linked L-fucose, R ² CH (COOH) ₂ .

1c 4. A compound of formula 1d, which is characterized in that

X oxygen,

Y NHCH ₂ CH ₂ OH,

R ¹ α1- (CH ₂ ) ₃ -glycosidically linked L-fucose,

R ^{2 means} (CH ₂ ) _n COOH and n = 1.

1 d

5. A compound of formula 1 e, which is characterized in that X 2 hydrogen atoms, Y OH,

R α1- (CH ₂ ) ₃ -glycosidically linked L-fucose, R ² (CH ₂ ) _n COOH and n = 1.

H

1e 6. A compound of formula 1f, which is characterized in that

X oxygen,

Y OH,

R ¹ α1-CH ₂ -glycosidically linked D-mannose,

R ^{2 means} (CH ₂ ) _n COOH and n = 1.

1f

7. A compound of formula 1g, which is characterized in that X is oxygen, Y OH,

R ¹ denotes 1- (CH ₂ ) ₂ -glycosidically linked L-fucose, R ² (CH ₂ ) _n COOH and n = 1.

ig The object stated at the outset is also achieved by a process for the preparation of a compound of the formula I, which is characterized in that one of the two OH groups of a commercially available 3,5-dihydroxy-benzoic acid ester is treated with an electrophilic, OH-protected fucose or mannose block selectively O-alkylated in the 3-position. The 5-OH position of the intermediate is then further alkylated or acylated, the ester function in the 1-position is modified if necessary and finally the end product of the formula I is released by splitting off the protective groups.

Conversely, it is also possible to alkylate or acylate only in the 3-position and only then to introduce the sugar unit in the 5-position by alkylation. Both processes lead to the same end product of formula I, and for each compound to be specially prepared, the optimal sequence of these 2 alternative synthesis strategies must be checked depending on the structure of the radicals R ¹ and R ² . In general, the former variant always leads to the desired end products, since the protected sugar units in the radical R ^{1 to} be introduced cannot be further alkylated or acylated. The reverse procedure according to the second variant is advantageous if there are no further alkylatable or acylatable functions, in particular basic N atoms, in the radical R ^{2 to} be introduced, so that the more valuable sugar component is only used later in the synthesis sequence.

Formula scheme 3 illustrates the synthesis of compounds of formula I according to the first variant and formula scheme 4 gives an example of the synthesis sequence according to the second variant.

95% 83%

65% 98%

32%

Formula scheme 3

The individual steps of the synthesis sequences can be carried out according to standard methods of organic synthesis familiar to the person skilled in the art. Components 2-4 shown in FIG. 5 are used as synthesis building blocks are in turn obtainable by methods known from the literature. For example, fucosides 2 and 3 can still be found in bioorgan. Med.Chem.Lett. 1996, 4, 1149-1165. The synthesis of the C-mannoside 4, which has not yet been described, can be carried out in a multi-stage process as described in FIG. The primary alcohols 2a, 3a or 4a can be used directly for the alkylation of the phenolic OH group, for example with

98%

12 11

100%

Mitsonobu reaction can be used. Alternatively, only the reactive bromides 2b, 3b and 4b are prepared from the alcohols with CBrCI ₂ CBrCI ₂ or CBr ₄ / triphenylphosphine / / triethylamine in dichloromethane, which in turn are used as alkylation reagents in the presence of bases for ether formation.

3a: X = OH 4a: X = OH 3b: X = Br 4b: X = Br

Formula scheme 5

After introducing a radical R ² into a compound of the formula I, differentiation of the acid functions in the radical R ² and in the 1 position of the benzene ring can be achieved by means of suitable protective groups. For example, even the sterically hindered tert-butyl group in the glycol side chain is removed more alkaline than the aromatic ester group on the way to the synthesis of 1d (formula scheme 4). The latter can therefore be easily converted into the amides in question simply by heating with nucleophilic amines. On the other hand, the aromatic methyl ester group can be more easily reduced to the hydroxyl group than the tert-butyl ester in the side chain of a radical R ² in I by sterically hindered hydride reagents such as DIBAL or Li (s-butyl) ₃ AIH (example compound 1e). In the last two synthesis steps, the benzyl protective groups are removed hydrogenolytically with hydrogen over a palladium catalyst and the ester groups are saponified in an alkaline manner. The sequence of these two steps can also be interchanged. The end compounds of formula I can be used as free acids or in the form of their salts with organic or inorganic bases for the biological and pharmacological Investigations and applications are used.

The α-C-mannoside 4a shown in formula scheme 6, which is required as an intermediate for the synthesis of the compounds I according to the invention, is new. To produce 4a, commercially available methylmannoside 14 is converted to 15, the anomeric OMe group is first exchanged for an O-acetyl group (compound 16) and then a cyanide residue in the 1 position is introduced using trimethylsilylcyanide / boron trifluoride etherate. The product mainly contains the preferred α-C anomer of the formula 17, which is saponified to give the methyl ester 18, which in turn is converted to the primary alcohol 4a.

Accordingly, the present invention also relates to the provision of the interconnect 4a.

Compound 4a can then optionally be processed according to standard procedures, e.g. with triphenylphosphine / dibromotetrachloroethane or tetrabromomethane) are converted into the reactive bromide 4b, which can be used for the alkylation of the 3- or 5-OH groups in 3,5-dihydroxybenzoic acids.

14 15 16

TMSCN BF ₃ OEt

17

50% α

Formula scheme 6 (22% ß) The affinity of the compounds of the formula I according to the invention for selectins can be checked using the cell adhesion assays described below.

Primary assays to investigate the effect of the compounds according to the present invention on cell attachment to recombinant, soluble selectin fusion proteins.

In order to test the effectiveness of the compounds according to the invention on the interaction between the E- and P-selectins (old nomenclature ELAM-1 or GMP-140) with their ligands, an assay is used which is only specific for one of these interactions. The ligands are offered in their natural form as surface structures on promyelocytic HL60 cells. Since HL60 cells have ligands and adhesion molecules of different specificity, the desired specificity of the assay can only be achieved via the binding partner. Genetically engineered soluble fusion proteins from the respective extracytoplasmic domain of E- or P-selectin and the constant region of a human immunoglobulin of the IgG1 subclass were used as binding partners.

Production of E-selectin (Lx)

The genetic construct "ELAM-Rg" published by Walz et al., 1990, was used to produce soluble E-selectin (Lx) fusion protein. For expression, the plasmid DNA was transfected into COS-7 cells (ATCC) using DEAE-dextran (molecular biological methods: see Ausubel, FM, Brent, R., Kingston, RE, Moore, DD, Seidman, JG, Struhl, K. and Smith, JA 1990. Current Protocols in Molecular Biology, John Wiley, New York). Seven days after the transfection, the culture supernatant is obtained, freed from cells and cell fragments by centrifugation and brought to 25 mM Hepes pH 7.0, 0.3 mM PMSF, 0.02% sodium azide and stored at +4 C. (Walz, G., Aruffo, A., Kolanus, W., Bevilacqua, M. and Seed, B. 1990. Recognition by ELAM-1 of the sialyl-Lex determinant on myeloid and tumor celis. Science 250, 1 132-1135.)

Production of P-selectin-IgG1

The genetic construct "CD62Rg", published by Aruffo et al., 1991, is used to produce the soluble P-selectin-IgG1 fusion protein. The further procedure corresponds to the production of E-selectin (Lx) shown under A1.

Aruffo, A., Kolanus, W., Walz, G., Fredman, P. and Seed, B. 1991. CD62 / -P-Selectin recognition of myeloid and tumor cell sulfatides. Cell 67, 35-44.

Production of CD4-IgG1

The genetic construct "CD4: IgG1 hinge" published by Zettlemeissl et al., 1990 is used to produce the soluble CD4-IgG1 fusion protein. The further procedure corresponds to the production of E-selectin (Lx) shown under A1. (Zettelmeissl, G., Gregersen, J.-P., Duport, JM, Mehdi, S., Reiner, G. and Seed, B. 1990. Expression and characterization of human CD4: Immunoglobulin Fusion Proteins. DNA and Cell Biology 9 , 347-353.)

Implementation of the HL60 cell adhesion assay on recombinant, soluble adhesion molecules

1. 96-well microtiter test plates (Nunc Maxisorb) are incubated with 100 μl of a goat anti-human IgG antibody (Sigma) diluted in 50 mM Tris pH 9.5 (1 + 100) for 2 hours at room temperature. After removing the antibody solution, it is washed once with PBS. 2. 150 μl of the blocking buffer are left in the wells for 1 hour at room temperature. The composition of the blocking buffer is: 0.1% gelatin, 1% BSA, 5% calf serum, 0.2 mM PMSF, 0.02% sodium azide. After the blocking buffer has been removed, it is washed once with PBS.

3. 100 μl of cell culture supernatant from appropriately transfected and expressing COS cells are pipetted into the wells. Incubation takes place for 2 hours at room temperature. After removing the cell culture supernatant, it is washed once with PBS.

4. 20 μl of binding buffer are added to the wells. The binding buffer has the composition: 50 mM Hepes, pH 7.5; 100 mM NaCl; 1 mg / ml BSA; 2 mM MgCl ₂ ; 1 mM CaCl ₂ ; 3 mM MnCl ₂ ; 0.02% sodium azide; 0.2 mM PMSF. 5 μl of the test substance are pipetted in, mixed by swirling the plate and incubated for 10 minutes at room temperature.

5. 50 ml of an HL60 cell culture with 200,000 cells / ml are centrifuged at 350 g for 4 min. The pellet is resuspended in 10 ml RPMI 1640 and the cells centrifuged again. To label the cells, 50 μg BCECF-AM (Molecular Probes) are dissolved in 5 μl anhydrous DMSO; 1.5 ml of RPMI 1640 are then added to the BCECF-AM / DMSO solution. The cells are resuspended with this solution and incubated at 37 ° C. for 30 minutes. After centrifugation at 350 g for two minutes, the labeled cell pellet is resuspended in 11 ml of binding buffer and the resuspended cells are distributed in 100 μl aliquots in the microtiter plate wells. The plate is left to stand at room temperature for 10 minutes to allow the cells to sediment on the bottom of the test plate. The cells have the opportunity to adhere to the coated plastic.

6. To stop the test, completely immerse the microtiter plate in the stop buffer at a 45-degree angle (25 mM Tris, pH 7.5; 125 mM NaCl; 0.1% BSA; 2 mM MgCl ₂ ; 1 mM CaCl ₂ ; 3mM MnCl2; 0.02% sodium azide). The stop buffer is removed from the wells by inverting and the procedure is repeated twice.

7. The BCECF-AM-labeled cells adhering to the well are measured in a cytofluorimeter (Millipore) with a sensitivity setting of 4, an excitation wavelength of 485/22 nm and an emission wavelength of 530/25 nm.

Leukocyte adhesion - testing the effectiveness of the compounds according to the invention in vivo (intravital microscopy in the rat):

In inflammatory processes and other conditions that activate the cytokines, tissue destruction by immigrant or microcirculation blocking leukocytes plays a crucial role. The first phase, which is decisive for the further disease process, is the activation of leukocytes within the bloodstream, especially in the pre- and postcapillary area. After the leukocytes have left the axial flow of the blood, the leukocytes first attach to the inner wall of the vessel, i.e. on the vascular endothelium. All subsequent leukocyte effects, i.e. the active migration through the vessel wall and the subsequent oriented migration in the tissue are subsequent reactions (Harlan, J.M., Leukocyte-endothelial interaction, Blood 65, 513-525, 1985).

This receptor-mediated interaction of leukocytes and endothelial cells is considered an initial sign of the inflammatory process. In addition to the adhesion molecules that have already been expressed physiologically, inflammatory mediators (leukotrienes, PAF) and cytokines (TNF-alpha, interleukins) cause massive, time-graded expression of adhesion molecules on the cells. They are currently divided into three groups: 1. immunoglobulin gene superfamily, 2. integrins and 3. selectins. While the adhesion between Molecules of the Ig gene superfamily and the protein-protein bonds are in the foreground in the cooperation between selectins lectin-carbohydrate bonds (Springer, TA, Adhesion receptors of the immune system. Nature 346, 425-434, 1990; Huges, G., Cell adhesion molecules - the key to an universal panacea, Scrips Magazine 6, 30-33, 1993; Springer, TA, Traffic Signals for lymphocyte recirculation and leukocyte emigration; The multistep paradigm. Cell 76, 301-314, 1994) .

Method:

The induced adhesion of leukocytes is quantified using an intravital microscopic examination technique in the rat mesentery (Atherton A. and Born GVR, Quantitative investigations of the adhesiveness of circulating polymorphnuclear leukocytes to blood vessel walls. J. Physiol. 222, 447-474, 1972; Seiffge , D. Methods for the investigation of the receptor-mediated interaction between leukocytes and endothelial cells in the inflammatory process, in: Replacement and supplementary methods for animal experiments in biomedical research, SchöffI, H. et al., (Ed.) Springer, 1995 (in press) ). Inhaled anesthesia is induced by an intramuscular injection of urethane (1.25 mg / kg body weight). After free preparation of vessels (femoral vein for the injection of substances and carotid artery for measuring blood pressure), catheters are integrated into them. The corresponding transparent tissue (mesentery) is then exposed according to the standard methods known in the literature and stored on the microscope stage and covered with 37 C paraffin oil (Menger, MD and Lehr, H., A. Scope and perspetives of intravital microscopy-bridge over from in vitro to in vivo, Immunology Today 14, 519-522, 1993). The test substance is administered iv to the animal (10 mg / kg). The experimental increase in blood cell adhesion is triggered by systemic administration of lipopolysaccharide (LPS, 15 mg / kg) 15 minutes after application from test substance by cytokine activation (Foster SJ, Mc Cormick LM, Ntoiosi BA and Campbell D., Production of TNF -alpha by LPS-stimulated murine, rat and human blood and its pharmacological modulation, Agents and Actions 38, C77-C79, 1993, 18.01.1995). The resulting increased adhesion of leukocytes to the endothelium is quantified directly under the vital microscope or with the help of fluorescent dyes. All measuring processes are recorded by a video camera and saved on a video recorder. Over a period of 60 minutes, the number of rolling leukocytes (ie all visible rolling leukocytes that are slower than the flowing erythrocytes) and the number of adhering leukocytes on the endothelium (residence time longer than 5 seconds) are recorded every 10 minutes. After the end of the experiment, the anesthetized animals are euthanized without pain by systemic injection of T61. For evaluation, the results of 8 treated and 8 untreated animals (control group) are compared (the results are given in percent).

Reperfusion model to investigate the influence of neutrophil adhesion in the course of ischemia / reperfusion on the open rabbit heart:

The hearts are perfused at constant pressure using the Langendorff technique with nutrient solution and with / without leukocytes or active ingredient. Then ischemia is caused by the prevention of left coronary artery (30 min). After reperfusion (30 min), the leukocyte accumulation is evaluated histologically. In the course of the test, potentials and arrhythmias are also measured on 256 electrodes (total test duration approx. 90 min). In 6 of 7 untreated hearts perfused with leukocytes, pronounced arrhythmias occur due to leukocyte infiltration, while the hearts treated with an active ingredient (RGDS peptides, chondroitin suifate) develop reduced leukocyte accumulation and arrhythmias. The investigated compound 3a was highly effective in the range of approximately 1 μM (strong reduction in arrhythmias).

Because of their valuable pharmacological properties, the compounds according to the present invention and their physiologically tolerable salts are very suitable for use as medicinal products in mammals, in particular humans. The present invention therefore further relates to a medicament comprising at least one compound of the formula I and the use thereof for the manufacture of a medicament for the therapy or prophylaxis of diseases which are associated with excessive cell adhesion mediated by selectin receptors in the tissue affected by the disease, for example rheumatism , Cardiovascular diseases such as reperfusion injuries, ischemia or heart attack.

The medicinal products are particularly suitable for the treatment of acute and chronic inflammations which can be characterized pathophysiologically by a disturbance in the cell circulation, for example by lymphocytes, monocytes and neutrophilic granulocytes. These include autoimmune diseases such as acute polyarthritis, rheumatoid arthritis and insulin-dependent diabetes (diabetes mellitus IDDM) acute and chronic graft rejection, shock lung (ARDS, adult respiratory distress syndrome), inflammatory and allergic skin diseases such as psoriasis and contact eczema, cardiac Circulatory diseases such as myocardial infarction, reperfusion injuries after thrombolysis, angioplasty or by-pass surgery, septic shock and systemic shock. Another potential indication is the treatment of metastatic tumors, because tumor cells carry surface antigens that have both sialyl-Lewis-X and sialyl-Lewis-A structures as recognition epitopes. In addition, these drugs, which are stable in the acidic environment of the stomach, can also be used in combination with antibiotics for the anti-adhesive therapy of Helicobacter pylori and related microorganisms. Therapy of the cerebral form of malaria is also conceivable with the help of these drugs.

The medicaments according to the invention are generally administered intravenously, orally or parenterally or as implants, but rectal use is also possible in principle. Suitable solid or liquid pharmaceutical preparation forms are, for example, granules, powders, tablets, dragées, (micro) capsules, suppositories, syrups, emulsions, suspensions, aerosols, drops or injectables Solutions in ampoule form and preparations with protracted release of active ingredient, in the production of which carriers and additives and / or auxiliaries such as explosives, binders, coating agents, swelling agents, lubricants or lubricants, flavorings, sweeteners or solubilizers are used. Examples of commonly used carriers or auxiliaries are magnesium carbonate, titanium dioxide, lactose, mannitol and other sugars, talc, milk protein, gelatin, starch, vitamins, cellulose and their derivatives, animal and vegetable oils, polyethylene glycols and solvents, such as sterile water, alcohols , Called glycerin and polyhydric alcohols.

The pharmaceutical preparations are preferably produced and administered in dosage units. Fixed dosage units are tablets, capsules and suppositories.

Different daily doses are necessary for the treatment of a patient depending on the effectiveness of the compound, the type of application, the type and severity of the disease, the age and body weight of the patient. Under certain circumstances, however, higher or lower daily doses may also be appropriate. The daily dose can be administered either by single administration in the form of a single dosage unit or else several small dosage units or by multiple doses divided at certain intervals. The daily dose to be administered may also depend on the number of receptors expressed during the course of the disease. It is conceivable that only a few receptors are expressed on the cell surface in the initial stage of the disease and, consequently, the daily dose to be administered is lower than in the case of severely diseased patients.

The medicaments according to the invention are produced by bringing a compound according to the present invention into a suitable administration form with conventional carriers and, if appropriate, additives and / or auxiliaries. It is also conceivable to use compounds of the formula I for the preparation of an agent for diagnosing a disease which is associated with excessive cell adhesion mediated by selectin receptors in the tissue affected by the disease.

Examples of the preparation of the compounds according to the invention:

Example 1: Synthesis of Compound 6:

1,434 g (2.66 mmol) of 3- (tri-0-benzyl-α-L-fucopyranosyl) propyl bromide and 1.34 g (7.98 mmol) of methyl 3,5-dihydroxybenzoate (5) are mixed with 0.553 g (3.99 mmol) of potassium carbonate stirred in 20 ml DMF at 0 ° C for 12 h and then at 20 ° C for 24 h. After adding 50 ml of saturated ammonium chloride solution, the mixture is shaken out several times with ethyl acetate (EA), the EA phases are washed with saturated sodium hydrogen carbonate solution, dried and concentrated. 1.03 g (61.4%) of product 6 crystallize from toluene / EA.

¹ H NMR (300 MHz, δ [ppm] rel. TMS, CDCI ₃ ): 7.23-7.37 (m, Phe, 15H), 7.10 / 7.13 (2 dd, ArH-2, ArH-5, 2H), 6.56 (t, ArH-4, 1 H), 5.33 (s, OH, 1 H), 4.50-4.80 (m, H-benzyl, 6H), 3.88 (s, OMe), 3.70-4.10 (m, FucH-17 -27-37-47-5 ^' , OCH ₂ CH ₂ CH ₂ , OMe, 10H), 1.60-1.90 (m, OCH ₂ CΗ ₂ CH ₂ , 4H), 1.26 (d, Fuc-6 ^' , 3H).

Example 2: Synthesis of compound 7:

0.839 g (1.34 mmol) of compound 6 and 0.247 ml (2.68 mmol) of methyl bromoacetate are stirred for 5 h at 50 ° C. with 0.555 g (4.02 mmol) of potassium carbonate in 10 ml of DMF. After adding 50 ml of saturated ammonium chloride solution, the mixture is shaken out repeatedly with (EE), the EE phases are washed with saturated sodium hydrogen carbonate solution, dried and concentrated. Column chromatography on silica gel 60 in hexane / EA (3/1) yields 0.78 g (83.5%) of product 7. ¹ H-NMR (300 MHz, δ [ppm] rel. TMS, CDCI ₃ ): 7.23-7.37 (m, Phe , 15H), 7.22 / 7.14 (2 dd, ArH-2, ArH-5, 2H), 6.67 (t, ArH-4, 1 H), 4.50-4.80 (m, H-benzyl, OCH ₂ C0 ₂ Me, 8H), 3.88 (s, 1-C0 ₂ Me), 3.80 (s, OCH ₂ C0 ₂ Me), (3.70-4.20 (m, FucH- 17-27-37-47-5 ', OCH ₂ CH ₂ CH ₂ , OMe, 13H), 1.60-1.92 (m, OCH ₂ CH ₂ CH ₂ , 4H), 1.26 (d, Fuc-6 ', 3H).

Example 3: Synthesis of Compound 8:

0.782 g (1.12 mmol) of compound 7 are dissolved in 15 ml of dry methanol and 0.5 ml of acetic acid and hydrogenated at 20 ° C. over 0.50 g of 10% PD / C. After filtering off the catalyst and removing the solvents, 0.400 g (83%) of compound 8 are obtained. H-NMR (300 MHz, δ [ppm] rel. TMS, CD ₃ OD): 7.21 / 7.13 (2 dd, ArH-2 , ArH-5, 2H), 6.72 (t, ArH-4, 1 H), 4.73 (s, OCH ₂ C0 ₂ Me, 2H), 3.90 (s, 1-C0 ₂ Me), 3.81 (s, OCH ₂ C0 ₂ Me), (3.65 -4.10 (m, FucH- 17-27-37-47-5 ^' , OCH ₂ CH ₂ CH ₂ , 2xOMe, 13H), 1.60-1.92 (m, OCH ₂ CH ₂ CH_ ₂ , 4H), 1.22 (d, Fuc-6 ^' , 3H).

Example 4: Synthesis of Compound 1a:

0.400 mg (0.935 mmol) of compound 8 are stirred with 5 ml of 2N NaOH at 20 ° C for 0.5 h. The solution obtained is neutralized with 1 N HCl (pH 5.73), filtered through a 0.45 μm filter and concentrated. After medium-pressure chromatography on RP (reversed phase), 0.409 g (98%) of the end compound 1a is obtained as a DC-uniform (EE / MeOH / H ₂ 0 / AcOH 5/3/1 / 0.5) disodium salt of the formula C ₁₈ H ₂₂ O ₁₀ Na ₂ (444.3g / mol); H-NMR (300 MHz, δ [ppm], D ₂ 0): 7.17 / 7.05 (2 dd, ArH-2, ArH-5, 2H), 6.70 (t, ArH-4, 1 H), 4.79 (s , OCH ₂ C0 ₂ H, 2H), 4.10-4.25 (m, OCH ₂ , 2H), 3.93 -4.04 and 3.74-3.88 (2 m, 2H or 3H, FucH-17-27-37-47-5 ^' ), 1.65-1.97 (m, OCH ₂ CH ₂ CH ₂ , 4H), 1.11 (d, Fuc-6 ^' , 3H).

¹³ C NMR (75.4 MHz, APT, δ [ppm], D ₂ 0): 176.48, 174.55 (C = 0), 158.90, 158.73 ArC-3 / -5), 138.98 (ArC-1), 108.24, 107.96 (ArC-2 / -6), 104.39 (ArC-4), 75.68, 71.81, 69.77, 67.99, 66.78 (FucC-17-27-37-47-5 ^' ), 68.23 (OCH ₂ CH ₂ CH ₂ ), 66.91 (OCH ₂ CO ₂ H), 24.77, 19.74 (CH ₂ CH ₂ ), 15.57 (C-6 ^' _Fuc ). Example 5: Synthesis of Compound 9:

0.690 g (1.10 mmol) of compound 6, 0.245 g (1.21 mmol) of 4-nitrophenyl chloroformate, 15 mg (0.12 mmol) of 4-dimethylaminopyridine (DMAP) and 0.17 ml (1.21 mmol) of triethylamine are dissolved in 10 ml of dry dichloromethane. After stirring at 0 ° C. for 5 h, 0.47 ml (2.76 mmol) of N-ethyl-N, N-diisoprpylamine (DIPEA) and 0.187 ml (1.21 mmol) of piperidine-4-carboxylic acid ethyl ester are added. After stirring at 0 ° C. for 12 h, the mixture is diluted with 30 ml of dichloromethane and washed 3 times with saturated sodium bicarbonate solution and water and dried. After concentrating the organic phase, 0.81 g of crude product is obtained, which is purified using the Biotage-Flash-40 system (cartridge Flash 40 M, 90 g silica gel 32-63 μ, eluent 600 ml EA / hexane 1/4, 800 ml EA / Hexane 1/3, 600 ml EE / Hexane 1/2). Yield product 9: 0.845 g (94.6%).

¹ H-NMR (300 MHz, δ [ppm] rel. TMS, CDCI ₃ ): 7.23-7.42 (m, Phe, ArH-2, ArH-5, 17H), 6.85 (t, ArH-4, 1 H) , 4.49-4.80 (m, H-benzyl, 6H), 4.17 (q, C0 ₂ CH ₂ CH ₃ ), 2H), 3.88 (s, C0 ₂ Me), 3.78-4.05 (m, FucH-17-27- 37-47-5 ^' , OCH ₂ CH ₂ CH ₂ , OMe, 10H), 1.60-2.04 (m, OCH ₂ CH ₂ CH ₂ , 4H), 1.28 (t, CO, CH CH _3. 3H), 1.26 ( d, Fuc-6 ^' , 3H).

Example 6: Synthesis of Compound 1b:

0.835 g (1.03 mmol) of compound 9 are dissolved in 30 ml of dry methanol and 0.5 ml of acetic acid and hydrogenated at 20.degree. C. over 0.30 g of 10% PD / C. After filtering off the catalyst, stripping off the solvents and column chromatography on silica gel 60 (dichloromethane / methanol 17/1), 0.365 g (65%) of the degenerated intermediate is obtained, which is stirred with 5 ml of 1N NaOH at 0 ° C. for 12 h . The solution obtained is neutralized with 1 N HCl (pH 3), filtered through a 0.45 μm filter and concentrated. After medium pressure chromatography on RP material (methanol / water 1/1), 0.110 g (32%) of end product 1b of the formula C ^ H ^ NO ^ (497.5) is obtained (without optimization);

¹ H-NMR (300 MHz, δ [ppm], D ₂ 0): 7.37 / 7.22 (2 dd, ArH-2, ArH-5, 2H), 6.90 (t, ArH- 4, 1 H), 3.71- 4.35 (FucH- 17-27-37-47-5 ^' , 2xCH ₂ CHHN, OCH ₂ , 9H), 2.90-3.20 (2xCH ₂ CHHN, 2H), 2.36-2.50 (tt, CH ₂ CHC0 ₂ H, 1 H), 1.52-2.20 (m, OCH ₂ CH ₂ CH ₂ , CH ₂ CHC0 ₂ H, 8H), 1.13 (d, Fuc-6 ^' , 3H). ³ C NMR (75.4 MHz, APT, δ [ppm], D ₂ 0): 183.94 (pip-C = 0), 173.78 (ArC = 0), 158.61, 155.18, 151.59 (ArC-3 / -5, NC = 0), 139.01 (ArC-1), 114.96, 112.96, 112.42 (ArC-2 / -6 / -4), 75.63, 71.78, 69.76, 67.96, 66.76 (FucC-17-27-37-47-5 ^' ), 68.38 (OCH ₂ ), 43.91 (CCH ₂ CH ₂ N), 43.97 (CCH ₂ CH ₂ N), 28.53 (CCH ₂ CH ₂ N), 24.74, 19.73 (OCH ₂ CH ₂ CH ₂ ), 15.57 (C -6 ^' _Fuc ).

Example 7: Synthesis of Compound 1c:

0.493 g (0.788 mmol) of compound 6 and 0.172 ml (0.87 mmol) of bromomonic acid diethyl ester are stirred with 0.10 g (1.2 mmol) of sodium hydrogen carbonate in 20 ml of DMF at 0 ° C. for 48 h. After adding 50 ml of saturated ammonium chloride solution, the mixture is shaken out repeatedly with (EE), the EE phases are washed with saturated sodium hydrogen carbonate solution, dried and concentrated. Column chromatography on silica gel 60 (40-63 μ) in hexane / EA (3.5 / 1) yields 0.31 g (50%) of the intermediate C ₄₅ H ₅₂ 0 ₁₂ (784.9), which is further reacted as follows:

0.30 g (0.38 mmol) of the intermediate compound are dissolved in 10 ml of dry methanol and 0.4 ml of acetic acid and hydrogenated at 20 ° C. over 0.12 g of 10% PD / C. After filtering off the catalyst and removing the solvents, 0.15 g (75%) of the debenzylated intermediate is obtained, which is further reacted as follows:

0.148 mg (0.287 mmol) of the intermediate are stirred with 1.5 ml of 2N NaOH at 20 ° C for 2 h. The solution obtained is neutralized with 1 N HCl (pH 2) and concentrated. After MPLC purification on RP material (methanol / water), 0.090 g (68%) of end compound 1c is obtained as a TLC-uniform (EE / MeOH / H ₂ 0 / AcOH 5/3/1 / 0.5) product of the formula C ₂₀ H ₂₆ O ₁₂ (458.4);

¹ H-NMR (300 MHz, δ [ppm], D ₂ 0): 7.13 / 7.07 (2 m, ArH-2, ArH-5, 2H), 6.77 (m, ArH- 4, 1 H), 3.70 - 4.30 (m, FucH-17-27-37-47-5 ^' , OCH ₂ ), 1.65-2.00 (m, OCH ₂ CH ₂ CH ₂ , H), 1.11 (d, Fuc-6 ^' , 3H). ³ C-NMR (75.4 MHz, APT, δ [ppm], D ₂ 0): 158.90, 158.18 (ArC-3 / -5), 138.59 (ArC-1), 108.45, 108.36 (ArC-2 / -6) , 104.86 (ArC-4), 75.65, 71.81, 69.74 _r 67.97, 66.75 (FucC- 17-27-37-47-5 ^' ), 68.23 (OCH ₂ ), 24.74, 19.73 (CH ₂ CH ₂ ), 15.56 ( C-6 ^' _Fuc ).

Example 8: Synthesis of Compound 10:

5.74 g (34.1 mmol) of methyl 3,5-dihydroxybenzoate (5) and 3.78 ml of tert-butyl bromoacetate (addition in portions) are stirred with 7.08 g (25.6 mmol) of potassium carbonate in 60 ml of DMF at 0 ° C. for 12 h . After adding 200 ml of saturated ammonium chloride solution, the mixture is shaken out several times with diethyl ether, the ether phases are washed with saturated sodium bicarbonate solution, dried and concentrated. 5.64 g (58% with respect to 5; 78% with respect to benzoic acid ester) crystallize from toluene / EA (7/1) product 10. ¹ H-NMR (300 MHz, δ [ppm] rel. TMS, CDCI ₃ ): 7.17 / 7.07 ( 2 dd, ArH-2, ArH-5, 2H), 6.67 (t, ArH-4, 1 H), 6.30 (s, OH, 1 H), 4.54 (s, OCH ₂ ), 2H), 3.88 (s , OMe), 1.50 (s, tert-Bu, 9H).

Example 9: Synthesis of Compound 11:

0.885 g (1.64 mmol) of 3- (tri-0-benzyl-α-L-fucopyranosyl) propyl bromide and 0.695 g (2.46 mmol) of compound 10 are mixed with 0.51 g (3.69 mmol) of potassium carbonate in 10 ml of DMF for 12 h at 20 ° C and then stirred at 60 ° C for 7 h. After adding 50 ml of saturated ammonium chloride solution, the mixture is shaken out repeatedly with EA, the EA phases are washed with saturated sodium hydrogen carbonate solution, dried and concentrated. After column chromatography on silica gel 60 (EA / hexane 5/1), 1.20 g (98% based on Fuc. Component) of product 11 are obtained. ¹ H-NMR (300 MHz, δ [ppm] rel. TMS, CDCI ₃ ): 7.24-7.38 (m, Phe, 15H), 7.20 / 7.13 (2 dd, ArH-2, ArH-5, 2H), 6.67 (t, ArH-4, 1 H), 4.52 (s, OCH ₂ C0 ₂ tBu), 4.50-4.80 (m, H-benzyl, OCH ₂ C0 ₂ tBu, 8H), 3.89 (s, OMe), 3.70-4.10 (m, FucH- 17-27-37-47-5 ^' , CH ₂ CH ₂ CH ₂ , OMe, 10H), 1.60-1.90 (m, OCH ₂ CH ₂ CH ₂ , 4H), 1.49 (s, tBu, 9H), 1.25 (d, Fuc-6 ^' , 3H).

Example 10: Synthesis of Compound 12:

0.901 mg (1.217 mmol) of compound 11 are stirred for 12 h at 0 ° C in 20 ml of methanol and 3 ml of 1N NaOH. After adding 1N HCl (pH 50-5.5) and 200 ml of EA, the organic phase is washed with dilute aqueous citric acid solution, dried and concentrated. After column chromatography on silica gel 60 (EA / hexane 1/1 with 0.1% AcOH), 0.48 g (57.6%) of compound 12 are obtained. ¹ H-NMR (300 MHz, δ [ppm] rel. TMS, CDCI ₃ ): 7.26 -7.38 (m, Phe, 15H), 7.23 / 7.15 (2 dd, ArH-2, ArH-5, 2H), 6.68 (t, ArH-4, 1 H), 4.67 (s, OCH ₂ C0 ₂ H) , 4.50-4.80 (m, H-benzyl, OCH ₂ C0 ₂ H, 8H), 3.89 (s, OMe), 3.70-4.10 (m, FucH-17-27-37-47-5 ^' , OCH ₂ CH ₂ CH ₂ , OMe, 10H), 1.60-1.90 (m, OCH ₂ CH ₂ CH ₂ , 4H), 1.26 (d, Fuc-6 ^' , 3H).

Example 11: Synthesis of Compound 13:

0.462 g (0.675 mmol) of compound 12 are heated to 100 ° C. for 1 h with 7 ml of ethanolamine. The cooled solution is taken up in EA and washed with saturated NaCl solution. After drying and concentration, the mixture is filtered through silica gel (dichloromethane / methanol 10/1, 1% HOAc). 482 mg (100%) of compound 13 are obtained.

¹ H-NMR (300 MHz, δ [ppm] rel. TMS, CDCI ₃ ): 7.25-7.36 (m, Phe, 15H), 6.81 / 6.89 (2 m, ArH-2, ArH-5, 2H), 6.50 (m, ArH-4, 1 H), 4.50-4.80 (m, H-benzyl, OCH ₂ CO ₂ H, 8H), 3.70-4.10 (m, FucH-17-27-37-47-5 ^' , OCH ₂ CH ₂ CH ₂ , OCH ₂ CH ₂ N, 11 H), 1.60-1.90 (m, OCH ₂ CH ₂ CH ₂ , 4H), 1.25 (d, Fuc-6 ^' , 3H).

Example 12: Synthesis of Compound 1d:

0.484 g (0.678 mmol) of compound 13 in 6 ml of dioxane and 0.2 ml of acetic acid are hydrogenated at 20 ° C. over 0.200 g of 10% PD / C. After filtering off the catalyst and stripping off the solvent, the residue is dissolved in water. The sterile filtered The solution is freeze-dried and 0.213 g (70.7%) of the final compound 1d of the formula C ₂₀ H ₂₉ NO ₁₀ (443.5) is obtained;

¹ H NMR (300 MHz, δ [ppm], D ₂ 0): 6.93 / 6.87 (2 dd, ArH-2, ArH-5, 2H), 6.95 (t, ArH-

4, 1 H), 4.79 (s, OCH ₂ C0 ₂ H, 2H), 3.90 -4.10 (2m, OCH ₂ CH ₂ CH ₂ , CH ₂ OH, 4H), 3.72-

3.83 (2m, FucH-17-27-37-47-5 ^' , 5H), 3.51 (t, J = 6Hz, CH ₂ N, 2H), 1.68-1.94 (m,

OCH ₂ CH ₂ CH ₂ , 4H), 1.12 (d, J = 6Hz, Fuc-6 ^' , 3H).

¹³ C NMR (75.4 MHz, APT, δ [ppm], D ₂ 0): 174.57, 169.75 (2 C = 0), 159.37, 158.56

(ArC-3 / -5), 135.81 (ArC-1), 106.77, 105.92, 104.83 (ArC-2 / -6 / -4), 75.61, 71.74,

69.80, 67.92, 66.77 (FucC- 17-27-37-47-5 ^' ), 68.21 (OCH ₂ CH ₂ CH ₂ ), 65.75

(OCH ₂ CO ₂ H), 59.93 (CH ₂ OH), 42.01 (CH ₂ N), 24.84, 19.76 (CH ₂ CH ₂ ), 15.58 (C-6 ^' _Fuc ).

Example 13: Synthesis of (2,3,4,6-tetra-0-benzyl-α-D-mannopyranosyl) #yanide (17).

A solution of 20 g (103 mmol) of α-D-methyl- is added to a mixture of 17.3 g (576.8 mmol) of NaH (80% in mineral oil) and 63.6 ml (535.6 mmol) of benzyl bromide in DMF (325 ml). mannopyranoside in DMF (300 ml). After stirring at 20 ° C. for 18 h, methanol (13 ml) and water (1.25 l) are added. The mixture is extracted with diethyl ether, concentrated and chromatographed on silica gel (hexane / EA 5/1). 44 g (77%) of the tetrabenzylmannoside 15 are obtained.

The acetolysis of 22.5 g (40.6 mmol) of tetrabenzylmannoside 15th in acetic anhydride / acetic acid / sulfuric acid (340/590 / 2.3ml) provides after working up

1. ice water;

2. extraction with EE;

3. neutralization of the organic phase with sodium hydrogen carbonate;

4. Chromatography with hexane / EA 4/1) the anomeric acetate 16 in a yield of 20 g (76%).

8.8 g (15.12 mmol) of the anomeric acetate 16 are glycosylated on the anomeric carbon atom by reaction with 8.7 ml (69.55 mmol) of cyanotrimethylsilane in Acetonitrile (200 ml) under the catalysis of boron trifluoride-diethyl etherate complex.

After stirring for 15 min, the mixture is neutralized with aqueous sodium hydrogen carbonate and chromatographed on silica gel (hexane / EA 6/1).

Yield: 17 (α-anomer) 4.0 g (50%) and 17 (β-anomer) 1.75 g (22%). H-NMR (300 MHz, CDCI ₃ , rel. TMS, δ [ppm]): 3.69-4.11 (m, 2-H, 3-H, 4-H, 5-H, 6-

H _a, 6H _s, 6H); 4.78 (d, J _{1 2} = 2.3 Hz, 1-H _α , 1 H), 7.15-7.39 (m, 5Ph, 20H).

Example 14: Synthesis of C-1-hydroxymethyl-2,3,4,6-tetra-0-benzyl-α-D-mannopyranoside (4a).

4.6 g (8.38 mmol) of cyanide 17 (α) are dissolved in a 0.015 M sodium methanolate solution (838 ml). After stirring at 20 ° C. for 8 h, 1 M hydrochloric acid (100 ml) is added. Concentration and chromatography on silica gel (hexane / EA 5/1 to 4/1) yielded 2.9 g (91%, based on the reacted reactant) of the methyl ester 18 and 1.6 g of reactant 17.

To reduce the ester 18, 3.25 g (5.58 mmol) are dissolved in diethyl ether (85 ml) and added dropwise at 0 ° C. to a mixture of diethyether (43 ml) and 1 M ethereal LiAIH ₄ solution (5.75 ml). After stirring for 10 min, EA (20 ml) and then water (10 ml) are carefully added. It is diluted with ethyl acetate, the organic phase washed with brine and water, dried over sodium sulfate, filtered, concentrated and the residue on silica gel (hexane / EA 3/2 to 1/1). chromatographed. Yield of compound 4a: 2.7 g (87%). ¹ H-NMR (300 MHz, CDCI ₃ ): δ = 2.05 (bt, OH, 1 H), 3.64 (dd, 1 H), 3.70 (dd, 1 H), 3.95, 4.06 (2m, CH ₂ OH, 2H), 3.72-3.88 (m, 5H), 4.42-4.57 (m, 4CH ₂ Ph, 8H), 7.18-7.36 (m, 4Ph). Example 15: Inhibition of cell adhesion in vitro:

IC ₅₀ values for E-selectin [mM] and for P-selectin [mM]:

Example 16: Inhibition of leukocyte adhesion in vivo (intravital microscopy), application 3 mg / kg i.v.

Claims

Claims:

1. Compound of formula I.

in what mean

X oxygen, sulfur or 2 hydrogen atoms,

Y OH, NH ₂ , NHCH ₂ COOH, NHCH (COOH) ₂ , NHCH ₂ CH ₂ OH,

NHCH ₂ CH ₂ CH ₂ OH, NHCH ₂ CH ₂ CH ₂ CH ₂ OH, R ¹ C-glycosidically linked fucose or mannose, R ² (CH ₂ ) _n COOH, (CH ₂ ) _n SO ₃ H, (CH ₂ ) _n CH (COOH) ₂ , CH (COOH) ₂ , or

COR ³ , where n is an integer from 1 to 5 and R ^{3 is} an amino acid residue of the formulas NH (CH ₂ ) _m COOH, NH (CH ₂ ) _m SO ₃ H, or NHCH (R ⁴ ) COOH, pyrrolidine-2-carboxylic acid , Pyrrolidine-3-carboxylic acid, piperidine-2-carboxylic acid, piperidine-3-carboxylic acid or

Piperidine-4-carboxylic acid and where m is an integer from 1 to 5 and R ^{4 is} an amino acid side chain selected from the group of naturally occurring proteinogenic α-amino acids

Amino acid mean.

Compound according to claim 1, characterized in that the radical R ¹ in formula I. means.

3. A compound according to claim 1, characterized in that the radical R ¹ in formula I.

means.

4. A compound according to claim 1, characterized in that the rest

R ¹ in Formula I.

means.

5. A compound according to claim 2, characterized in that X is oxygen,

Y OH,

R ^{2 means} (CH ₂ ) _n COOH and n = 1.

6. A compound according to claim 2, characterized in that X is oxygen,

Y OH and

R ² COR ³ , where R ^{3 represents} a 1- (4-carboxy) piperidyl radical.

7. A compound according to claim 2, characterized in that X is oxygen,

Y OH and

R ^{2 represents} CH (COOH) ₂ .

8. A compound according to claim 2, characterized in that X is oxygen,

Y means NHCH ₂ CH ₂ OH, R ² (CH ₂ ) _n COOH and n 1.

9. A compound according to claim 2, characterized in that X 2 is hydrogen,

Y OH,

R ^{2 is} (CH ₂ ) _n COOH and n 1.

10. A compound according to claim 3, characterized in that X is oxygen,

Y OH,

R ^{2 is} (CH ₂ ) _n COOH and n 1.

11. A compound according to claim 4, characterized in that X is oxygen,

Y OH,

R ^{2 is} (CH ₂ ) _n COOH and n 1.

12. A process for the preparation of a compound of formula I according to any one of claims 1 to 1 1, characterized in that one of the two OH groups in 3,5-dihydroxybezoic acid esters with an OH-protected fucose or mannose building block selectively in the 3-position O-alkylated and then the 5-OH position of the intermediate further alkylated or acylated, the ester function in the 1 position hydrolyzed or coupled with amines and finally the end product of formula I is released by splitting off all protective groups.

13. A process for the preparation of a compound of formula I according to any one of claims 1 to 1 1, characterized in that one of the two OH groups in 3,5-dihydroxybezoic acid esters in the 3-OH position is alkylated or acylated and then with a OH-protected 1-C-fucose or 1-C-mannose building block in the 5-position O-alkylated, the ester function in the 1-position hydrolyzed or coupled to amines and finally the end product of the formula I is released by splitting off all protective groups.

14. The method according to claim 12 or 13 for the preparation of a compound of formula I according to claim 3 or 10, characterized in that the 1-C-mannose block is a compound of formula III

where OBn is an O-benzyl protecting group.

15 The method according to claim 14, characterized in that the

Compound of formula III according to claim 14 from a primary alcohol of formula II

prepared by first providing a methylmannoside with O-benzyl protecting groups, replacing the anomeric O-methyl group with an O-acetyl group, then substituting the said O-acetyl group for a cyanide residue in the 1-position, which mainly forms The α-C anomer of the nitrite thus obtained is separated off and saponified to form an ester, which is finally reduced to the primary alcohol of the formula II, after which the primary alcohol of the formula II is converted to the corresponding bromide.

16. A compound of formula II according to claim 15.

17. Medicament containing at least one compound according to one of claims 1 to 11.

18. Use of a compound according to any one of claims 1 to 11 for the manufacture of a medicament for the therapy or prophylaxis of a disease which is accompanied by an excessive cell adhesion mediated by selectin receptors in the tissue affected by the disease.

19. Use according to claim 18, characterized in that the disease is a cardiovascular disease.

20. Use according to claim 18, characterized in that the disease is an autoimmune disease such as rheumatoid arthritis or osteoarthritis.

21. Use of a compound according to any one of claims 1 to 11 for the Preparation of an agent for diagnosing a disease associated with excessive selectin-mediated cell adhesion in the tissue affected by the disease.