WO1998023626A1 - Derives d'acide benzoique anti-adhesifs - Google Patents

Derives d'acide benzoique anti-adhesifs Download PDF

Info

Publication number
WO1998023626A1
WO1998023626A1 PCT/EP1997/006308 EP9706308W WO9823626A1 WO 1998023626 A1 WO1998023626 A1 WO 1998023626A1 EP 9706308 W EP9706308 W EP 9706308W WO 9823626 A1 WO9823626 A1 WO 9823626A1
Authority
WO
WIPO (PCT)
Prior art keywords
formula
compound
cooh
compound according
disease
Prior art date
Application number
PCT/EP1997/006308
Other languages
German (de)
English (en)
Inventor
Gerhard Kretzschmar
Alexander Toepfer
Original Assignee
Glycorex Ab
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Glycorex Ab filed Critical Glycorex Ab
Publication of WO1998023626A1 publication Critical patent/WO1998023626A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H7/00—Compounds containing non-saccharide radicals linked to saccharide radicals by a carbon-to-carbon bond
    • C07H7/04—Carbocyclic radicals

Definitions

  • the invention relates to derivatives of benzoic acid in which ⁇ -C-fucosides or ⁇ -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).
  • receptors 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-1 ⁇ ) or tumor necrosis factor (TNF- ⁇ ), 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.
  • acute and chronic diseases are 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.
  • 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.
  • MI myocardial ischemia / infarction
  • psoriasis dermatitis
  • ARDS respiratory distress syndrome in adults
  • 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).
  • 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.
  • 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.
  • R 1 C-glycosidically linked fucose or mannose
  • R 2 (CH 2 ) ⁇ 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 ⁇ -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
  • R 2 COR 3 , where R 3 represents a 1- (4-carboxy) piperidyl radical.
  • R 1 denotes ⁇ 1- (CH 2 ) 3 -glycosidically linked L-fucose, R 2 CH (COOH) 2 .
  • a compound of formula 1 e which is characterized in that X 2 hydrogen atoms, Y OH,
  • 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.
  • 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.
  • 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
  • Mitsonobu reaction can be used.
  • 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.
  • 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).
  • sterically hindered hydride reagents such as DIBAL or Li (s-butyl) 3 AIH (example compound 1e).
  • 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 ⁇ -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.
  • 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 ⁇ -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.
  • 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.
  • 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.
  • E- and P-selectins old nomenclature ELAM-1 or GMP-140
  • 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.
  • EAM-Rg The genetic construct "ELAM-Rg” published by Walz et al., 1990, was used to produce soluble E-selectin (Lx) fusion protein.
  • Lx E-selectin
  • 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).
  • 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.
  • CD4 IgG1 hinge
  • 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.
  • Lx E-selectin
  • 96-well microtiter test plates (Nunc Maxisorb) are incubated with 100 ⁇ l 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 ⁇ l 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.
  • 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 ⁇ l of the test substance are pipetted in, mixed by swirling the plate and incubated for 10 minutes at room temperature.
  • the labeled cell pellet is resuspended in 11 ml of binding buffer and the resuspended cells are distributed in 100 ⁇ l 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.
  • 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.
  • 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.
  • inflammatory mediators leukotrienes, PAF
  • cytokines TNF-alpha, interleukins
  • 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.
  • 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).
  • LPS lipopolysaccharide
  • 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.
  • the number of rolling leukocytes ie all visible rolling leukocytes that are slower than the flowing erythrocytes
  • the number of adhering leukocytes on the endothelium are recorded every 10 minutes.
  • the anesthetized animals are euthanized without pain by systemic injection of T61.
  • the results of 8 treated and 8 untreated animals are compared (the results are given in percent).
  • 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 ⁇ M (strong reduction in arrhythmias).
  • 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.
  • 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.
  • 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.
  • 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.
  • 1,434 g (2.66 mmol) of 3- (tri-0-benzyl- ⁇ -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.
  • EA ethyl acetate
  • Example 16 Inhibition of leukocyte adhesion in vivo (intravital microscopy), application 3 mg / kg i.v.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention concerne des dérivés d'acide benzoïque où, en position 3 et 5 du composé cyclique aromatique, des α-C-fucosides ou des α-C-mannosides, ainsi que des chaînes ou des composés cycliques alkyle portant au moins une fonction acide ont été rattachés par l'intermédiaire de liaisons ether et de groupes séparateurs courts. L'invention concerne en outre un procédé permettant de préparer ces composés et leur utilisation pour préparer des médicaments et des agents diagnostiques.
PCT/EP1997/006308 1996-11-25 1997-11-12 Derives d'acide benzoique anti-adhesifs WO1998023626A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19648681.5 1996-11-25
DE1996148681 DE19648681A1 (de) 1996-11-25 1996-11-25 Antiadhäsive Benzoesäure-Derivate

Publications (1)

Publication Number Publication Date
WO1998023626A1 true WO1998023626A1 (fr) 1998-06-04

Family

ID=7812656

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP1997/006308 WO1998023626A1 (fr) 1996-11-25 1997-11-12 Derives d'acide benzoique anti-adhesifs

Country Status (2)

Country Link
DE (1) DE19648681A1 (fr)
WO (1) WO1998023626A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20210094573A (ko) * 2018-11-22 2021-07-29 이도르시아 파마슈티컬스 리미티드 클로스트리듐 디피실에 대한 안정성 백신

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
PHYTOCHEMISTRY, vol. 33, no. 4, 1993, pages 813 - 816, XP002060354 *

Also Published As

Publication number Publication date
DE19648681A1 (de) 1998-05-28

Similar Documents

Publication Publication Date Title
EP0787739B1 (fr) Anto-adhésive piperidine et pyrollidine-acides carboxyliques
DE19532902A1 (de) Neuartige Glycomimetika als Selektin-Antagonisten und daraus hergestellte entzündungshemmend wirkende Arzneimittel
EP0786467A1 (fr) Acides dicarboxyliques multi-fucosylés à propriétés anti-adhésives
EP0714903B1 (fr) Carbohydrat conjugués utilisés comme inhibiteurs d'adhésion cellulaire
EP1169313B1 (fr) Derives de 1,4-benzothiazepin-1,1-dioxyde substitues par des residus de sucre, leur procede de production, des medicaments contenant ces composes et leur utilisation
US6133240A (en) Tetrahydronapthalene derivatives and their therapeutic use
DE69627380T2 (de) Biphenyl-hydroxamat matrix-metalloproteine inhibitor
JP3004297B2 (ja) リゾスフィンゴ脂質誘導体
EP0671408A2 (fr) Mimétiques d'hydrates de carbone ayant des propriétés anti-adhésives
EP0671409A2 (fr) Dérivés de l'acide malonique ayant des propriétés anti-adhésives
MXPA96004657A (en) Acidos piperidina- and pirrolidina-carboxilicos antiadhesi
DE60215530T2 (de) Neues glykolipid, und heilmittel für autoimmunerkrankungen, das dieses als wirkstoff enthält
DE60011568T2 (de) Derivate von Monosacchariden als Zelladhäsionsinhibitoren
EP0147777A2 (fr) Dérivés d'amides d'acides carboxyliques M-glycosylés comme produit pour combattre les maladies de type rheumatismal
DE19540388A1 (de) Neuartige Glycomimetika als Selektin-Antagonisten und entzündungshemmend wirkende Arzneimittel
EP0741139A1 (fr) Esters de l'acide sulfurique d'aminosucres
DE3881862T2 (de) Disaccharidderivate.
DE69031448T2 (de) Halbsynthetische Gangliosidanaloge
WO1998023626A1 (fr) Derives d'acide benzoique anti-adhesifs
CA1329591C (fr) Produits pharmaceutiques et nouveaux composes de lactosyl et leur preparation
WO1998030573A1 (fr) Analogues anti-adhesifs de sulfatides
DE69908919T2 (de) Verfahren zur herstellung von gm3
EP0807437A1 (fr) Compositions contenant de polymeres polyacryliques et polymethacryliques ayant des propriétés antiadhesives
DE4025330A1 (de) Lipidderivate und verfahren zu deren herstellung
JPH10152498A (ja) 糖脂質誘導体、それを有効成分とする免疫抑制剤およびその製造用中間体

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): CA JP US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
NENP Non-entry into the national phase

Ref country code: CA

122 Ep: pct application non-entry in european phase